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Objective To explore the effect of hydroxy-carboxylic acid receptor 2(HCAR2) on erythroid differentiation of human chronic myeloid leukemia K562 cells under hypoxia. Methods The cells were cultured in medium containing hemin to compare the ability of K562 cells to differentiate into erythroid cells under normoxia/hypoxia conditions. Then the expression of HCAR2 was interfered with genetic methods to knockdown or overexpress it in K562 cells, respectively. Benzidine staining, RT-qPCR, Western blot and flow cytometry were used to detect the role of HCAR2 in the erythroid differentiation of K562 promoted by hypoxia. Results On day 1, 2, 3 after differentiation, the expression level of HCAR2 and the erythroid differentiation ability of K562 cells increased gradually with time, and hypoxia enhanced the above phenotypes compared with normoxia. In HCAR2-silenced K562 cells, erythroid cells was reduced and the expression of erythroid differentiation markers CD235a and γ-globin decreased. Simultaneously, the proportion of double-positive cells expressing erythroid cell surface molecules CD71 and CD235a was lower than that of the control group. On the contrary, the overexpression of HCAR2 in K562 cells increased erythroid cells generation as well as elevated the expression levels of CD235a and γ-globin. In addition, the proportion of CD71/CD235a double-positive cells also increased in HCAR2-overexpressed K562 cells. Conclusion HCAR2 mediates hypoxia in promoting the erythroid differentiation of K562 cells.

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Objective To investigate the protective effect and mechanism of Hudi Enteric capsules in ionizing radiation injury to small intestinal crypt cells(IEC-6 cells) in rats. Methods IEC-6 cells were irradiated with 6 mega electron volt X-rays(2, 4, 6, 8 and 10 Gy), cell clone formation assay was used to detect cell proliferation, and the 6 Gy ionizing radiation was selected to establish a cellular radiation damage model according to the cell survival rate. The effect of each concentration(12.5, 25, 50, 100 and 200 μg/ml) of Hudi enteric extract on the viability of IEC-6 cells was examined by cell counting kit-8(CCK-8) method, and the effect on the viability of IEC-6 cells after irradiation at(10, 20, 40 and 80 μg/ml) concentrations was examined according to the results.After obtaining the optimal irradiation dose and extract concentration, the cells were divided into control group, model group and Hudi extract group(80 μg/ml), the control group was pseudo-irradiated and the other two groups received 6 Gy of ionizing radiation, and the Hudi enteric extract group was pre-treated with drugs 2 h before irradiation. Apoptosis was detected by Annexin V-PI double staining; cell senescence was detected by β-galactosidase(β-Gal) staining; reactive oxygen species was detected by DCFH-DA fluorescent probe; the corresponding protein expression of p16, p21, Catalase(CAT) and Superoxide dismutase(SOD2) was detected by Western blot.Results The proliferation of IEC-6 cells was inhibited by radiation doses ranging from 4 Gy to 10 Gy(P<0.001);the concentrations of 40 and 80 μg/ml of Hudi enteric extract increased the survival rate of irradiated IEC-6 cells(P<0.001), and the apoptosis rate, β-Gal positivity rate and DCFH-DA fluorescence intensity in the Hudi enteric extract group were lower than those in the model group(P<0.05). The protein expressions of CAT and SOD2 in the Hudi extract group were higher than those in the model group(P<0.05), and the protein expressions of p16 and p21 were lower than those in the model group(P<0.05).Conclusion The mechanism of action of Hudi enteric capsules that attenuate radiation damage in IEC-6 cells may be related to the inhibition of reactive oxygen species production, reduction of oxidative stress, and attenuation of cellular senescence and apoptosis.

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Objective To analyze the genome structure and genetic characteristics of IncpGRT1plasmids fromPseudomonas, and elucidate its potential transmission risk. Methods The genomic DNA of the clinical isolate 15420352 was extracted after purification and preservation of the strain, and then the whole genome was sequenced, and then the type of the plasmid was identified. Sequence annotation and comparison of the backbone region and the accessory modules were performed on all five same type sequenced plasmids, including one plasmid p420352-strA in this study and four from GenBank. The plasmids were annotated byRAST,Plasmidfinder,Blast,ResFinder, andISfinder. The ORFs of the plasmid were annotated and drug resistance genes were found. Results All five plasmids were classified as new IncpGRT1type plasmids. The IncpGRT1backbone genes or gene loci were in all five plasmids, and they contained an auxiliary replicon besides the primary IncpGRT1replicon. Five IncpGRT1plasmids carried at least three different accessory modules, including thesrpregion, themsrregion, and a Tn5053family transposon. Three resistance genesstrA,strB, andmerwere obtained in these plasmids, which were involved in resistance to two categories of antibiotics and heavy metals. We also found that these plasmids carried at least one virulence genemsrand five key transporterssrp,emrE,mod,phn,andlpt, which could improve the environmental adaptability of the strains. Conclusion The IncpGRT1plasmids have become the important vector for the accumulation and spread of some drug resistance genes and virulence genes inPseudomonas, and have improved the environmental adaptability of the strain.

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Objective To investigate the partial mechanism and effect of transplanting with human umbilical cord-derived mesenchymal stem cells(hUC-MSCs) to repair articular cartilage defects in a rat model. Methods Critical-sized osteochondral defects were created in the trochlear grooves of rat femurs. The hUC-MSCs were transplanted into the defect of experimental knees. Sham group and model group knees were transplanted with saline. The effects of articular cartilage repair were evaluated at 10 weeks after surgery with International Cartilage Repair Society(ICRS) repair score, histological examination and immunohistochemical analysis.The effects of hUC-MSCs on the proliferation and migration of chondrocytes were detected by Transwellin vitro. Results After transplanting with hUC-MSCs, the articular surfaces of the defect sites were changed smoother thanthose of the model group, and the cellular morphology and arrangement were also improved in Safranin O staining or Masson staining results. Similar to surrounding normal articular cartilage tissue after treatment for 10 weeks, and the ICRS repair score was significantly elevated. In addition, hUC-MSCs treatment could promote chondrocytes proliferation, increase the expression of type Ⅱ(Col Ⅱ) collagen and decrease the level of type Ⅰcollagen(Col Ⅰ) in the articular defect site.Meanwhile, the increased protein expression of SOX9 and protein transportation to the nucleus were observed after treatment with hUC-MSCs for 10 weeks.In vitro, chondrocytes could be proliferated and migrated after co-cultured with hUC-MSCs. Conclusion Transplanting with hUC-MSCs can promote cartilage repair, and its role maybe related to the protection of the cartilage matrix and the promotion of chondrocyte proliferation and migration.

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Objective To investigate the effect of hypoxia/cold exposure on white fat browning in an obese rat model. Methods Obese rat model was constructed by high-fat feeding and randomly divided into control group, hypoxic group, low-temperature group and low-temperature hypoxic group, with 10 rats in each group. The changes of body weight and body fat were observed, and the morphological changes of rat adipose tissue cells were observed using HE staining and compared with the area of adipocytes. The expression of peroxisome proliferator-activated receptor γ2(PPAR-γ2), PR structural domain binding factor 16(PRDM16) and uncoupling protein 1(UCP-1) gene and the expression of UCP-1 protein in the adipose tissue of rats were examined by immunofluorescence, real-time fluorescence quantitative PCR and Western blot. Results Compared with the control group, the body weight and body fat of rats in the hypoxic, hypothermic and cryogenic hypoxic groups were reduced, and the body weight and body fat of rats in the hypothermic hypoxic group were lower than those in the hypoxic and cryogenic groups(P<0.05). Compared with the control group, the adipocyte area of rats in the hypoxia, hypothermia and low-temperature hypoxia groups was reduced, and the adipocyte area of rats in the low-temperature hypoxia group was lower than that in the hypoxia and low-temperature groups(P<0.05). Compared with the control group, the expression of PPAR-γ2, PRDM16 and UCP-1 genes all increased in the scapular brown adipose tissue(F=378.495, 102.061, 322.443,P<0.05) and decreased in the perirenal white adipose tissue(F=4.555,P<0.05) in the hypoxic, hypothermic and low-temperature hypoxic groups of rats, PRDM16 and UCP-1 gene expression all increased(F=24.387, 163.660,P<0.05). Compared with the control group, the expression of UCP-1 protein in scapular brown adipose tissue and perirenal white adipose tissue of rats in the hypoxic, hypothermic and low-temperature hypoxic groups increased(P<0.05); UCP-1 protein expression was lower in scapular brown adipose tissue and perirenal white adipose tissue of rats in the hypoxic and hypothermic groups compared to the hypothermic hypoxic group(P<0.05). Conclusion Hypoxia/cold exposure can induce white adipose browning and affect the body weight of rats by modulating the intra-adipose PPAR-γ2 and PRDM16 pathways which lead to high UCP-1 expression.

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Objective To investigate the effect of electroacupuncture(EA) treatment at pinch points(EX-B2) on motor function and Notch signaling pathway after spinal cord injury(SCI). Methods Seventy-two Sprague-Dawley(SD) male rats weighing(250±20) g were randomly divided into sham operation group, SCI, electroacupuncture group(SCI+EA) and acupuncture group(SCI+AP), with 18 rats in each group. A rat acute SCI model of T10 was established by the modified Allen′s method. The SCI+EA and SCI+AP groups received 15 minutes of electro acupuncture or acupuncture treatment per day. The motor function of the hind limbs of rats was assessed by Basso, Beattie and Bresnahan(BBB) scoring method, the pathological recovery changes of spinal cord tissues were observed by Hematoxylin-eosin(HE) staining, and the mRNA and protein expression levels ofHes 3,Notch 3andNotch 4were detected by real-time quantitative PCR and estern blot method, respectively, after 3, 7 and 14 days of intervention. The expression of glial fibrillary acidic protein(GFAP) was detected by immunohistochemistry after 14 days of intervention. Results Compared with the sham group, the BBB scores were reduced in rats after SCI surgery and there was significant hemorrhage, structural destruction and degeneration of spinal cord tissue(P<0.05), while the SCI+EA and SCI+AP groups were milder than the SCI group(P<0.05). The mRNA expression levels and protein expression levels ofHes 3,Notch 3andNotch 4as well as the expression levels of GFAP appeared significantly higher in the SCI group compared with the sham group(P<0.05), while the SCI+EA group appeared lower compared with the SCI group(P<0.05). Conclusion EA of EX-B2 can improve the locomotor function of rats with SCI, which may be related to inhibiting the expression of GFAP and the activation of Notch signaling pathway.

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Objective High performance liquid chromatography-mass spectrometry(LC-MS/MS) system was used to accurately determine 22 bile acids in serum, liver, amniotic fluid and placenta of pregnant mice, and a LC-MS/MS method was established for efficient detection and analysis of bile acids in serum, liver, amniotic fluid and placenta of mice. Methods Pregnant mice serum, liver, amniotic fluid and placenta samples were processed, with 0.1% glacial acetic acid in 4 mmol/L ammonium acetate aqueous solution as mobile phase A and pure methanol as mobile phase B, the flow rate was 0.4 ml/min, a gradient elution program was used to elute with Phenomenex Gemini 3 μm NX-C18 110A(100 mm × 2.0 mm) chromatographic column elution, and mass spectrometry detection system used an electrospray ion source for negative ion multiple reaction monitoring. Results The linear relationship of 22 bile acids in the quantitative range was good. The RSD of inter-day and intra-day precision at low, medium and high concentrations was 0.5%-7.4%, the matrix effect was 88%-110%, and the extraction recovery was 84%-108%. Conclusion In this experiment, LC-MS/MS was established to detect 22 bile acids in serum, liver, amniotic fluid and placenta of pregnant mice. The method not only has high sensitivity and selectivity, but also can stably detect a large number of samples.

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Objective To investigate the effect and mechanism of celecoxib on retinal vascular endothelial growth factor(VEGF) in rats with diabetic retinopathy. Methods Forty-five SD rats were divided into normal control group(NC),diabetic retinopathy group(DR), celecoxib intervention diabetic retinopathy group(DR+C). The diabetic model was established by intraperitoneal injection of 1% STZ. After one month, celecoxib(50 mg/kg) was given intragastric administration(1/day) in the DR+C group.Two months later, serum total cholesterol(TC) and insulin were detected.The histopathological changes of the retina were observed.The expression of junctional adhesion molecule-like protein(JAML),VEGF and their signal pathway proteins and the distributions of interleukin-10(IL-10), vascular cell adhesion molecules-1(VCAM-1) were detected by Western blot. HUVEC cells were divided into normal glucose group(NG),high glucose group(HG) and high glucose plus celecoxib group(HG+C) to detect the expression of the above proteins. Results Compared with DR,retina in DR+C group was thinner. The retina in the DR+C group was thicker than that in the NC group. The levels of retinal JAML,phosphatidylinositol kinase3(PI3K),phosphorylphosphatidylinositol kinase3(P-PI3K), hypoxia-inducible factor1-α(HIF1-α),and VEGF in DR+C group were lower than those in DR group, while higher than those in NC group.The expression of retinal IL-10 and VCAM-1 decreased.The content of TC in DR+C and DR group was higher than those in NC group(P<0.01), while the content of insulin in DR+C and DR group was lower than thlse in NC group(P<0.001). Compared with HG group, the expressions of JAML,PI3K,P-PI3K,HIF1-α,VEGF in HG+C group decreased, but was higher than those in NG group.There was no significant difference in PI3K among the three groups. Conclusion Celecoxib can decrease the expression of VEGF,IL-10, VCAM-1 in retina of DR rats, which may be related to the PI3K/HIF1-α signaling pathway mediated by JAML.

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Objective To explore the molecular mechanisms of immunosuppressive activity of soluble CD83(sCD83). Methods Bone marrow-derived dendritic cells(DCs) and CD4+T cells in the spleen were isolated from bone marrow progenitor cells of 6-8-week-old C57B/6 mice. DCs and CD4+T cells were treated with 10 μl PBS(Control group) or 10 μl 10 μg/ml soluble CD83(sCD83 group), respectively. Immature DCs(imDCs) in DCs and CD4+CD25+FOXP3+T cells(Tregcells) in CD4+T cells were determined by flow cytometry. Western blot was used to determine the expression levels of indoleamine 2,3-dioxygenase(IDO) in imDCs and the expres-sion levels of phosphatase and tensin homolog(PTEN), phosphorylated-Akt serine/threonine kinase 1(p-Akt), Akt, nucleus NF-κB subunit(RelB), cytoplasm RelB and programmed cell death 1(PD-1) in Tregcells. ELISA was used to determine the level of kynurenine in DCs culture supernatant, and the levels of interleukin-10(IL-10), transforming growth factor-β(TGF-β) and IL-12 in imDCs and CD4+T cells co-culture system. Results Compared with Control group, the percentage of DCs cell counts expressing the surface markers of CD40, CD83, CD86 and major histocompatibility complex class-Ⅱ(MHC Ⅱ) in sCD83 group decreased(P<0.05), with the level of kynurenine in DCs culture supernatant increased(P<0.05). Compared with Control group, the percentage of Tregcell counts in sCD83 group increased(P<0.05), with the expression levels of PTEN and cytoplasm RelB enhanced(P<0.05), while the expression levels of p-Akt, nucleus RelB and PD-1 decreased(P<0.05). There was no significant difference in the expression level of Akt between the two groups. In the imDCs and CD4+T cells co-cultured supernatants, compared with Control group, the levels of IL-10 and TGF-β increased, and the level of IL-12 decreased in sCD83 group(P<0.05). Conclusion sCD83 can inhibit CD4+T cells nucleus RelB levels and regulate cytokines secretion so as to have an effects on cellular crosstalk of imDCs and Tregcells and promote imDCs and Tregcells proliferation.

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Objective To explore the effects of an ultradian light cycle on the periodic expression of glutamate receptors in the suprachiasmatic nucleus and the lateral habenula nucleus. Methods An ultradian light cycle T7(3.5 hours/3.5 hours light: dark) was used to establish an ultradian light cycle model group and T24(12 hours/12 hours light: dark) was used to establish control group. The expression of key proteins in the brain regions of the suprachiasmatic nucleus(SCN) and the lateral habenula(LHB) of the hypothalamus were analyzed by Western blot. Including glutaminergic receptors, pituitary adenylate cyclase activating polypeptide(PACAP) receptors, and downstream signaling molecules. Results Western blot results showed that the expression of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor subunit 2(GluR2) in SCN under Zeitgeber time(ZT) 1 and ZT 5 increased under an ultradian light cycle compared with normal photoperiod(P<0.05,P<0.01), the overall expression of GluR2 in T7 group was higher than that in T24 group(P<0.01), the overall expression of GluR2 in LHB group T7 was lower than that in T24 group(P<0.01). Compared with T24 photoperiod, the overall expression of N-methyl-D-aspartic acid receptor subunit 2(NR2B) in SCN increased under T7 photoperiod(P<0.05), the overall expression of phosphorylation of extracellular signal-related kinase(P-ERK) in LHB significantly increased under T7 photoperiod(P<0.05). Conclusion An ultradian light cycle would cause an up-regulating of GluR2 and NR2B expression, a down-regulating of GluR2 expression and up-regulating P-ERK expression in LHB.

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Objective To explore the inhibitory effect of microRNA-200b(miR-200b) on pulmonary fibrosis in silicotic mice. Methods Male C57BL/6 mice were randomly divided into control group, SiO2group, SiO2+agomir-NC group and SiO2+miR-200b agomir group, with 6 mice in each group. The mice in the SiO2group were instilled intratracheally with 50 μl of saline containing 5 mg SiO2particles. miR-200b agomir and agomir-NC were delivered to mice in the SiO2+miR-200b agomir and SiO2+agomir-NC groups respectively by intratracheal instillation on day 1 and 7 after SiO2exposure. Animals were sacrificed after the lung function test on the 14th day. The pathological changes of lung tissues were observed by HE and Masson staining. Then the concentration of total proteins and the numbers of leukocytes in bronchoalveolar lavage fluid(BALF) were measured. The content of transforming growth factor-β1(TGF-β1) in BALF was detected by ELISA kit. Results Compared with the control group, inflammatory cell infiltration and collagen deposition were observed in the lungs tissues of the mice in the SiO2group and the SiO2+agomir-NC group, while these detrimental effects were weakened in the SiO2+miR-200b agomir group. In addition, compared with the control group, the frequency(f) was accelerated and the number of Pause(PAU) increased(P<0.05), while the ratio rate of achieving peak expiatory flow(Rpef) decreased in the SiO2group and the SiO2+agomir-NC group(P<0.05). After miR-200b agomir intervention, the lung function of silicotic mice was improved. Moreover, miR-200b agomir intervention could also effectively alleviate the increase in the concentrations of total protein, the numbers of total leukocytes and the secretion of TGF-β1 in the BALF of mice induced by SiO2stimulation. Conclusion miR-200b can effectively ameliorate the lung function of silicotic mice and inhibit the progress of fibrosis.

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Objective To investigate the role of early inhibition of Toll-like receptor 4(TLR4) in regulating hippocampal neuroimmune responseto adolescent ratsafter neonatal hypoxic-ischemic brain damage(HIBD). Methods Postnatal day 7 rats were randomized into controlgroup, hypoxic ischemia(HI)group, and HI+TAK-242(the specific inhibitor of TLR4)(TAK-242) group. The expression of TLR4 in rat hippocampus was detected by immuno-histochemistry at 3 days after HI. Immunofluorescence were used to determine the number of Iba-1+, GFAP+,CD161+, MPO+and CD3+cells in the hippocampus at 21 days after HI. Immunohistochemistry was used to detect ICAM-1 and C3 a expression in the hippocampal CA1 region; and Western blot was used to detect tumor necrosis factor interleukin IL-1β, TNF-α and IL-10 expression.Results Compared with control group, significantly raised TLR4 expression was observed in the left hippocampal CA1, CA3 and DG regions(P<0.01 or P<0.05), while the expression in the TAK-242 group lowered compared to the HI group(P<0.05). The number of GFAP+cells in the CA1 area of the hippocampus in the TAK-242 group of neonatal rats decreased compared to which in the HI group at 21 days after HI(P<0.05),but the number of CD3+T lymphocytes in the hippocampal CA1 area of new born rats in the HI group increased compared to which in the Control group(P<0.05), but the difference between TAK-242 and the Control group was not statistically significant. The number of Iba-1+cells, MPO+cells, CD161+cells, the expression of ICAM-1 and C3a in hippocampal CA1 region, and the expression of TNF-α, IL-1β and IL-10 in hippocampus of rats were not different among groups at 21 days after HIBD.Conclusion Early inhibition of TLR4 may ameliorate adolescent neuroimmune disorders by reducing the increase of hippocampal astrocytesafter neonatal HIBD.

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Objective To investigate the effects of carnosine(CAR) on streptozotocin(STZ) induced renal ferroptosis and inflammation in diabetic mice. Methods Type 1 diabetes mice model were induced by STZ, and normal C57 mice were used as normal control group. The C57 mice were divided into 5 groups(6-8 mice in each group): normal control group(NC), normal control + carnosine group(NC+CAR), STZ model group(STZ), STZ model + carnosine group(STZ+CAR), STZ model + ferroptosis inhibitor group(STZ+Fer-1). After feeding the mice for 16 weeks, serum samples were collected to detect blood creatinine(CRE) and urea nitrogen(BUN) levels. The urine of mice was collected to detect the 24-hour urinary albumin level. HE staining and PAS staining were performed to observe the degree of renal pathological injury. Real-time PCR was used to detect the expression of interleukin(IL)-1β, IL-6,monocyte chemotactic protein 1(MCP-1) and tumor necrosis factor-α(TNF-α) in mouse kidney tissue. The expression of reactive oxygen species(ROS) in mouse kidney was detected by immunofluorescence.Morphology of renal mitochondria was observed by transmission electron microscopy. The protein expression levels of glutathione peroxidase 4(GPX 4) and long-chain lipoacyl-CoA synthetase 4(ACSL4), which are ferroptosis indexes, were detected by Western blot. The contents of malondialdehyde(MDA), glutathione(GSH) and Fe2+in mouse kidney tissue were determined. Results Compared with NC group, CRE and BUN levels in STZ group increased(P<0.001); and ompared with STZ group, these indexes decreased in STZ+CAR group(P<0.001,P<0.01). Renal histopathological examination showed that compared with NC group, renal tubule dilatation, inflammatory cell infiltration and glycogen deposition significantly increased in STZ group; and compared with STZ group, tubule dilatation, inflammatory cell infiltration and glycogen deposition decreased in STZ+CAR group. Electron microscope results showed that the renal mitochondria in STZ group were swollen, membrane density increased, mitochondrial ridges decreased or absent, renal tubule dilation was improved significantly in the STZ+CAR group and STZ+Fer-1 group, and inflammatory cell infiltration was reduced. Real-time PCR test results showed that compared with NC group, mRNA expression levels of inflammatory factor(IL-1β, IL-6, MCP-1 and TNF-α) increased in STZ group(P<0.001 orP<0.01); and mRNA expressions of IL-1β, IL-6, MCP-1 and TNF-α were decreased in STZ+CAR group compared with STZ group(P<0.01 orP<0.05). Immunofluorescence results showed that compared with NC group, ROS level in kidney tissue of mice in STZ group increased(P<0.001); and compared with STZ group, the expression of ROS in kidney tissue of STZ+CAR group decreased while ROS expression in STZ+CAR group decreased(P<0.01). Compared with NC group, GPX4 expression and GSH content in kidney of STZ group decreased(P<0.001), and ACSL4 protein expression and MDA and Fe2+contents increased(P<0.01 orP<0.001), GPX4 expression and GSH content increased(P<0.01), ACSL4 protein expression and MDA and Fe2+content decreased in STZ+CAR group(P<0.01 orP<0.001). Conclusion CAR inhibits ferroptosis and inflammation in the kidney in diabetic mice induced by STZ, and improved renal pathological injury in diabetic mice.

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Objective To investigate the protective effect of dulaglutide on acute kidney injury(AKI) induced by lipopolysaccharide(LPS).Methods Twenty-four male C57BL/6 mice were randomly divided into Control group(normal saline), LPS group(LPS 15 mg/kg), LPS+Dul group(LPS 15 mg/kg+Dulaglutide 0.6 mg/kg) and Dul group(Dulaglutide 0. 6 mg/kg) with 6 mice in each group. The drug was administered by intraperitoneal injection. After drug intervention for 24 h, the body weight and kidney weight of mice were recorded, and kidney tissue and serum samples were collected. The pathological changes in kidney tissue were observed by HE staining.The serum urea nitrogen(BUN) and creatinine(CRE) levels were detected by the kit. The levels of cytokines interleukin(IL-6), tumor necrosis factor(TNF-α) and IL-1β in the kidney were detected by qRT-PCR. The contents of macrophage marker F4/80 and myeloperoxidase(MPO) in kidney were determined by immunohistochemistry.Results Compared with Control group, mice in LPS group lost weight and increased kidney weight(P<0.001). Moreover, the levels of BUN and CRE increased(P<0.001,P<0.01). Meanwhile, the mRNA levels of IL-6, IL-1β and TNF-α increased(P<0.05). There was obvious pathological damage in kidney tissue. In addition, macrophage and neutrophil infiltration increased in LPS group(P<0.001). Compared with LPS group,mice in LPS + Dul group gained weight and lost kidney weight(P<0.05,P<0.001). Moreover, the levels of BUN and CRE in LPS + Dul group decreased(P<0.01). The renal histological scores were reduced(P<0.05).In addition, the levels of IL-6, IL-1β and TNF-α in kidney tissue decreased(P<0.05 or P<0.01). Moreover,the infiltration of macrophages and neutrophils in kidney was reduced(P<0.01).Conclusion Dulaglutide has a protective effect on LPS-induced sepsis AKI, which may be related to reduce the expression of inflammatory mediators and decrease the infiltration of inflammatory cell.

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Objective To study the role of Toll-like receptor 7(TLR7) in trigeminal nerve pain in SD rats, and to explore the mechanism of TLR7 mediating related inflammatory factors through the activation of NF-κB pathway during the process of pain. Methods The rat model of trigeminal neuralgia(TN) was established by infraorbital nerve constriction injury(ION-CCI). The expression of TLR7 in trigeminal ganglion(TG) was detected by qRT-PCR and Western blot. Hydroxychloroquine(HCQ), an inhibitor of TLR7, was given to ION-CCI rats by intragastric administration. The expression of TLR7 and its downstream signal pathway NF-κB subunit p65, p-p65 and inflammatory factors(TNF-α, IL-1β) in TG were detected. Results After trigeminal nerve injury induced by ligation of infraorbital nerve, the expression of TLR7 in TG of rats significantly increased. After administration of TLR7 inhibitor, the expression of TLR7, TNF-α and IL-1β in TG decreased, the translocation and phosphorylation of p65 nucleus decreased, the activation of NF-κB signal pathway was inhibited, and the mechanical pain induced by ION-CCI was relieved in male SD rats. Conclusion TLR7 in TG regulates neuropathic pain by activating NF-κB signal pathway in primary sensory neurons and mediating the expression of inflammatory cytokines TNF-α and IL-1β.

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Objective To explore the role of Golgi protein ACBD3 knockdown in ferroptosis. Methods ACBD3 knockdownviashRNA, combined with treatment of ferroptosis inducer RSL3, cell viability, iron levels, reactive oxygen species(ROS) levels, and lipid peroxide levels in different treatment groups were measured to comprehensively analyze the role of ACBD3 in ferroptosis. Results Compared to control cells, the levels of ROS increased in ACBD3 knockdown cells. The total iron content was significantly reduced(P<0.001), while the Fe2+concentration in ACBD3 knockdown cells increased compared to control cells(P<0.001). In the presence of ferroptosis inducer RSL3, compared to control cells, cell viability significantly decreased(P<0.001), and lipid peroxide greatly increased in ACBD3 knockdown cells(P<0.001). Conclusion Golgi protein ACBD3 knockdown in mammalian cells changes iron homeostasis and promotes ferroptosis.

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Objective To explore the role and molecular mechanism of circHIPK3 in the migration and invasion of nasopharyngeal carcinoma cells. Methods Firstly, we searched public databases and analyzed the expression level of circHIPK3 in nasopharyngeal carcinoma tissues and non-cancerous tissues in Gene Expression Database(GEO).Then, the circHIPK3 siRNA was synthesized and transfected into the NPC cell line 5-8F. After validate the efficiency of circHIPK3 knockdown, the proliferation, migration and invasion capacities of 5-8F cells were evaluated by CCK-8, wound healing and transwell assay. Meanwhile, the mRNA and protein expression of downstream genes, includingCDH1,CDH2,MMP2,MMP9andIL-6/STAT3were determined by RT-qPCR and Western blot. Finally, transcriptome sequencing technology was used to analyze the significantly differentially expressed genes after the interference of circHIPK3-siRNA in 5-8F cells, in order to find out the key genes mediating the oncogenesis role of circHIPK3. Results circHIPK3 expression significantly increased in nasopharyngeal carcinoma tissues compared with non-cancerous tissues(P<0.05).After siRNA-specific knockdown of circHIPK3 expression in 5-8F cells, the proliferation, scratch healing rate, and the number of migrated/invaded cells were significantly reduced(P<0.05,P<0.01,P<0.05 respectively). Meanwhile, the mRNA and protein levels ofCDH1significantly increased(P<0.001), but theCDH2,MMP2,MMP9,IL-6/STAT3mRNA and protein levels were significantly reduced(P<0.05). Further analysis by RNA-seq showed that the expression of AL645922.1, SCO2, AL136295.1 and ISY1-RAB43 was significantly up-regulated(P<0.05), and the expression of BIVM-ERCC5, FAM47E-STBD1, INO80B-WBP1, NAIP, C15orf38-AP3S2, BCL2L2-PABPN1 and SMIM11B was significantly down-regulated(P<0.05) in circHIPK3-siRNA transfected 5-8F cells. Conclusion circHIPK3 expression is significantly upregulated in nasopharyngeal carcinoma tissues. circHIPK3 promotes 5-8F cells proliferation, migration and invasion by regulating the expression of E-Cadherin, N-Cadherin, MMP2, MMP9 and IL-6/STAT3. Genes such as AL645922.1 and BIVM-ERCC5 may play a key role in promoting the migration and invasion of nasopharyngeal carcinoma 5-8F cells mediated by circHIPK3.

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Objective Using RNA sequencing data from The Cancer Genome Atlas(TCGA) to explore the expression, mechanism, and clinical significance ofKCTD7in hepatocellular carcinoma(HCC). Methods The RNA sequencing data and clinical information of HCC patients were downloaded from the TCGA database. Univariate and multivariate Cox regression analyses were used to explore the correlation between the expression level of theKCTD7gene in HCC and the clinical information and prognosis of patients. Gene Set Enrichment Analysis(GSEA) was used to predict possible pathways regulated by theKCTD7gene in HCC. Single-sample GSEA(ssGSEA) was used to compare immune infiltration betweenKCTD7high expression group and low expression group.KCTD7knockdown HCC cell lines were established to explore its function and possible mechanism in HCC regulation. Results KCTD7was highly expressed in HCC tissues compared to normal tissues(P<0.01). The overall survival rate of HCC patients with high expression ofKCTD7gene was worse(P<0.05). The expression level ofKCTD7was an independent risk factor affecting the overall survival of HCC patients. GSEA results showed that theKCTD7gene was related to cell cycle signaling pathways. In the tumor microenvironment, high expression of theKCTD7gene was positively correlated with activated CD4+T cells, central memory CD4+T cells, and natural killer cells. Knocking downKCTD7might inhibit HCC cell proliferation, impair cell cycle distribution, and promote apoptosis. Conclusion KCTD7gene is highly expressed in HCC and affects the prognostic survival of HCC patients. Knocking downKCTD7can inhibit the proliferation of HCC cells and promote apoptosis.

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Objective To investigate the effect of platelet particles on the extent of intestinal inflammation and intestinal mucosal permeability in mice with dextran sodium sulfate induced colitis. Methods The experiment was divided into four groups: normal control group(n=10, drinking sterile distilled water + intraperitoneal injection of 0.9% sodium chloride solution), PMPs group(n=10, drinking sterile distilled water + intraperitoneal injection of PMPs), DSS model group(n=10, drinking DSS solution + intraperitoneal injection of 0.9% sodium chloride solution), and experimental group(n=15, drinking DSS solution + intraperitoneal injection of PMPs).Peripheral blood-derived PMPs suspension was collected from inflammatory bowel disease(IBD) patients. A colitis model was constructed in mice by allowing them to freely drink a 5% DSS solution for 1 week, followed by continuous intraperitoneal injection of PMPs for 7 days. Disease activity index(DAI) scores was recorded daily and the severity of intestinal inflammation with histopathological scores(HI) was assessed by HE staining of colon samples at the end of the experiment. Myeloperoxidase(MPO), neutrophil elastase(NE), citrullinated histone H3(citH3), and free DNA levels were measured in colon homogenate, observe intestinal mucosal structure by transmission electron microscopy, and intestinal permeability was tested using fluorescein isothiocyanate-dextran(FITC-D). Results Compared with the normal control group, the colonic mucosa of mice in the PMPs group showed edema, severe destruction of epithelial structure, extensive aggregation of inflammatory cells, and increased overall HI score(P<0.01); the levels of inflammatory factors such as IL-1β and TNF-α in colonic tissue homogenates of mice in the PMPs group increased(P<0.05), and the expression of NETs increased(P<0.05); the plasma FITC-D level of mice in the PMPs group significantly increased(P<0.05), and the permeability of intestinal mucosa increased.Compared with the DSS group, the experimental group mice had higher plasma FITC-D levels(P<0.05) and more electron microscopic colonic epithelial damage.Conclusion PMPs induces NETs formation in mice, promotes colonic inflammation in mice, increases intestinal mucosal permeability and aggravates intestinal inflammation in mice with DSS colitis.

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Objective To investigate the role of dapagliflozin(DAPA) in ox-LDL-induced pyroptosis of human myeloid leukemia monocytes(THP-1) derived foam cells. Methods THP-1-derived foam cell pyroptosis model was constructed by ox-LDL-induced THP-1derived macrophages. The experimental groups were set as follows: the blank control group(NC), the ox-LDL group(ox-LDL), and the drug intervention group(ox-LDL+DAPA). Oil Red O staining was used to detect the foam cell levels of macrophages. The cell proliferation and toxicity assay kit was used to detect the effect of DAPA on foam cell viability. Hoechst 33342/propidium iodide(PI) double staining was used to detect THP-1 derived foam cell pyroptosis. Cell immunofluorescence double staining was used to detect the effect of DAPA on the expression of pyroptosis key factor Caspase-1 in foam cells. The activity of lactate dehydrogenase(LDH) in the cell culture medium was detected using a microplate enzyme-linked immunosorbent assay. qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of Nod-like receptor pyrin domain containing 3(NLRP3), cystein-containing aspartate-specific protease-1(Caspase-1),apoptosis-associatedspeck-like protein containing CARD(ASC),gasdermin-D(GSDMD), interleukin(IL)-18andIL-1β,respectively. Results The CCK-8 assay indicated that the optimal intervention concentration of DAPA was 10 μmol/L. Oil Red O staining confirmed the successful construction of the THP-1-derived foam cell pyroptosis model. Compared with the blank control group, the expression levels ofNLRP3,Caspase-1,ASC,GSDMD,IL-18,IL-1βmRNA and protein significantly increased in ox-LDL group(P<0.05), as well as the number of PI-positive cells and LDH activity(P<0.05), the fluorescence intensity of Caspase-1 and the number of redlipid droplets in the cytoplasm of the cells. However, these effects were significantly reversed after DAPA intervention in the ox-LDL+DAPA group(P<0.05). Conclusion DAPA inhibits ox-LDL-induced pyroptosis in THP-1-derived foam cells.

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Objective To investigate the mechanism of miRNAs in glycyrrhizic acid treatment of alcohol-induced liver injury in rats. Methods 45 male SD rats were randomly divided into glycyrrhizin group, model group and control group. The rats in glycyrrhizin group were given 56% liquor and glycyrrhizin, the rats in model group were given 56% liquor, and the rats in control group were given distilled water for 8 weeks.The blood was collected and the serum was separated by centrifugation to detect the levels of aspartate aminotransferase(AST) and alanine aminotransferase(ALT). RNA were extracted from liver tissues, miRNAs were detected by rat miRNA microarray, and their expression levels were analyzed. The miRNA target genes of differential miRNA were predicted. Gene Ontology(GO) and KEGG Pathway enrichment analysis were used to understand the function of differential miRNA, and the differential miRNA-mRNA-Pathway regulatory network was constructed using Cytoscape to further screen the regulatory key miRNA and key pathways. qRT-PCR was used to verify the expression of selected miRNA. Results Compared with the model group, the glycyrrhizin group could significantly improve the liver tissue lesions, and reduce the liver serum AST and ALT levels(P<0.05). Compared the microarray data of glycyrrhizin group and model group, a total of 13 differentially expressed miRNA were screened(P<0.05,fold change≥1.5), of which 10 were up-regulated and 3 were down-regulated. The GO classification annotation of differential miRNA target genes showed that differential miRNA were related to cell adhesion, antioxidant activity, metabolic process, biological process regulation, cell killing, immune system and other functions. The KEGG Pathway analysis of differential miRNA target genes showed that MAPK signaling pathway, mTOR signaling pathway, Ras signaling pathway, Hippo signaling pathway, PI3K-Akt signaling pathway, wnt signaling pathway, apoptosis and other signaling pathways might play an important regulatory role in the improvement of alcoholic liver injury by glycyrrhic acid. Conclusion This study established the miRNA expression profile of glycyrrhizin in the treatment of alcoholic liver injury in rats, suggested that miR-615, miR-107-3p and miR-292-5p might play an important role in the treatment of alcoholic liver injury by glycyrrhizin.

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Objective To analyze the data related to the clinical outcome of preimplantation genetic testing(PGT) for double frozen, double biopsied blastocysts and double frozen, once biopsied blastocysts, in order to expand the existing data and provide some guidance for the clinical value and safety of PGT for frozen-thawed embryos. Methods Retrospective analysis was made on the 38 PGT cycles of frozen-thawed blastocysts. According to the frequency of biopsy, cases in the study were divided into two groups: double frozen, double biopsy(DFDB) group and double frozen, single biopsy(DFSB) group. The freezing method was vitrification. Results There were 24 patients in DFDB group, 34 blastocysts were not diagnosed in the last PGT cycle, 32 blastocysts survived after thawing, and the survival rate of thawed blastocysts was 94.12%. After the second biopsy of these 32 blastocysts, genetic testing was performed, and all of them were definitely diagnosed, including 15 normal blastocysts(46.88%) and 17 abnormal blastocysts(53.13%).There were 14 patients in DFSB.The remaining 50 blastocysts in the last ICSI cycle were thawed and all blastocysts survived after thawing. Biopsy of these 50 blastocysts and genetic analysis showed that 47 blastocysts were diagnosed, including 9 normal blastocysts(18.00%), 28 abnormal blastocysts(56.00%), 10 mosaic blastocysts(20.00%), and 3 undiagnosed blastocysts(6.00%). In DFDB group and DFSB group, 8 patients and 5 patients transferred the normal blastocystswhich all survivedafter thawing. There were 5 clinical pregnancies and 3 clinical pregnancies, respectively. One healthy live birth was obtained respectively in each group. Conclusion Acceptable pregnancy rate can be obtained whatever DFSB or DFDB blastocyst, which is of clinical value.However, due to the small sample size, we need to expand the sample size to further explore its safety.

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Objective The differences of cognitive impairment between children with attention deficit hyperactivity disorder(ADHD) and normal children were compared, the influencing factors of cognitive impairment of children with ADHD were analyzed. Methods A total of 133 children with ADHD were selected as the ADHD group, and 117 normal children were recruited as the control group. The severity of the subjects′ clinical symptoms was assessed using the Swanson, Nolan,&Pelham Rating Scale-Fourth Edition(SNAP-IV) and the Weiss Functional Impairment Rating Scale. The degree of cognitive functional impairment of subjects was assessed using the MATRICS cognitive suite(MCBB), and the executive function impairment of the subjects was assessed by the Behavior Rating Inventory of Executive Function(BRIEF). The differences in cognitive functional impairment of the two groups were compared. The children with ADHD were further divided into three groups according to their clinical phenotype and age, respectively, and the differences of cognitive impairment among the three groups were compared. The influencing factors of the degree of cognitive impairment in children with ADHD were analyzed by multiple linear regression. The improvement of methylphenidate sustained-release tablets on cognitive and executive functional impairment in children with ADHD was observed. Results The scores of SNAP-IV, Weiss Functional Impairment Rating Scale, connection test and BRIEF of ADHD patients were significantly higher than those of normal controls(P<0.05). The scores of symbol coding test and maze test of ADHD patients were significantly lower than those of normal controls(P<0.05). The score of symbol encoding test in children with ADHD-HI was significantly higher than that in ADHD-I and ADHD-C groups(P<0.05), and the total BRIEF score of ADHD-C was significantly higher than that in ADHD-I and ADHD-HI groups(P<0.05). With the increase of age, the score of connection test of ADHD children gradually decreased, while the scores of symbol coding test and maze test gradually increased(P<0.05). The results of multiple linear regression analysis showed that age was the influencing factor of ADHD children′s score in the connection test, symbol coding test and maze test(P<0.05); the scores of SNAP-IV and Weiss Functional Impairment Rating Scale were the influencing factors of BRIEF score of ADHD children(P<0.05). After methylphenidate treatment, the scores of connection testand BRIEF significantly decreased(P<0.001), while the scores of symbol coding test and maze test significantly increased(P<0.001). Conclusion The younger the age and the more serious the clinical symptoms related to ADHD suggest that the cognitive impairment of children with ADHD is more serious. After methylphenidate treatment, the degree of cognitive and executive impairment in children with ADHD are improved.

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Objective To explore the differences of serum inflammatory cytokines and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains-2(Tie-2) levels between patients with first-episode schizophrenia and healthy people, and the correlation with clinical symptoms. Methods A total of 168 participants were recruited, including 86 patients with first-episode schizophrenia(patient group) and 82 healthy people(control group). Demographic data, Positive and Negative Symptom Scale(PANSS) and Brief Psychiatric Rating Scale(PANSS) were collected at baseline. Serum inflammatory cytokines and Tie-2 levels were determined by Meso Scale Discovery(MSD). Results Compared with the control group, the levels of serum interleukin(IL)-1β and IL-4 in the patient group increased(P<0.05), while the level of Tie-2 decreased(P<0.05). IL-1β level in the patient group was positively correlated with the total score of BPRS, the score of BPRS hostility factor and the score of PANSS positive scale factor(P<0.05). The positive score of PANSS and the total score of BPRS in the patient group had a positive effect on IL-1β level(P<0.05).PANSS negative scale factor score, general psychopathological scale factor score and total score of PANSS in patient group also had positive effects on Tie-2 level(P<0.05). IL-1β level in the patient group could effectively predict the severity of clinical symptoms in patients with first-episode schizophrenia at the critical level of 1.181 pg/L, with a specificity of 0.850 and a sensitivity of 0.972. Tie-2 level in the patient group can effectively predict the severity of clinical symptoms in patients with first-episode schizophrenia at the critical level of 2 127.076 pg/L, with specificity of 0.675 and sensitivity of 0.639. The AUC of IL-1β and Tie-2 were 0.836 1 and 0.646 2, respectively. Conclusion The levels of IL-1β, IL-4 and Tie-2 in patients with first-episode schizophrenia are different from those in healthy people. IL-1β levels in patients with first-episode schizophrenia are correlated with part of clinical symptoms. IL-1β and Tie-2 levels in the patient group may be influencing factors with high sensitivity and specificity in predicting the severity of clinical symptoms in patients with first-episode schizophrenia.

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Objective To investigate the feasibility and safety of transumbilical single-port laparoscopic total hysterectomy with a self-made single-port laparoscopic platform for uterine fibroids. Methods 114 patients with uterine fibroids who needed total hysterectomy were divided into 3 groups: Microchannel transumbilical single-port laparoscopy group(group Micro-channel,n=38), standard access transumbilical single-port laparoscopy group(group Single-port,n=25) and traditional laparoscopy group(group Traditional,n=51), in which self-made single-port laparoscopic access platform was used in group Micro-channel and group Single-port. The operation time, intraoperative blood loss, preoperative and postoperative hemoglobin value changes, postoperative VAS pain score, incision cosmetic score, postoperative complications, postoperative gastrointestinal recovery time, treatment costs and hospital stay were compared among the three groups. Results Compared with the other two groups, Microchannel transumbilical single-port laparoscopy group had less intraoperative blood loss, shorter hospital stay, faster postoperative recovery, lower postoperative VAS score and higher postoperative cosmetic score, and the differences were statistically significant(P<0.05). Transumbilical single-port laparoscopic surgery had shorter discharge time, lower pain scores, and better cosmetic scores than traditional laparoscopic surgery, and the differences were statistically significant(P<0.05).There was no significant difference in the incidence of postoperative complications, the rate of postoperative analgesic use and the change of postoperative hemoglobin among the three groups(P>0.05). Conclusion Microchannel transumbilical single-port laparoscopic surgery is safe and effective in the treatment of uterine fibroids, and can improve the therapeutic effect of patients. The self-made single-port laparoscopic platform is safe and effective, and can reduce the cost of treatment for patients.

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Objective To analyze the association between multiple loci of genes and the risk of early-onset preeclampsia(EOPE) with pregnancy induced hypertension. Methods Among 382 EOPE patients who had lived in Yanbian area for more than 10 years, 192 patients were randomly selected as case group.At the same time, 192 cases of natural delivery in the hospital were randomly selected as the control group.PCR-RFLP method was used to determine the specific genotype and allele distribution information, and non-conditional Logistic method was used to obtain odds ratio(OR) and 95% confidence interval(CI) to confirm the risk of various genotypes. Results There were two alleles of T and C at theGRB2rs8082005 locus, TC, TT, and CC genotypes. There were two alleles of T and C at theRXRArs3849222 locus, TC, TT, and CC genotypes. Through unconditional logistic regression analysis, in theGRB2rs8082005 locus, compared with the TT genotype, the CC genotype was more susceptible to EOPE(OR=3.155, 95%CI=1.513-6.579,P=0.002). In the explicit model, compared to patients with TT+TC genotype, patients with CC genotype increased the risk of EOPE(OR=3.000, 95%CI=1.495-6.022,P=0.002). In theRXRArs3849222 locus, compared with the CC genotype, the TT genotype was more susceptible to EOPE(OR=2.031, 95%CI=1.077-3.820,P=0.028). In the invisible model, compared to patients with CC+CT genotype, patients with TT genotype had an increased risk of EOPE(OR=2.549, 95%CI=1.421-4.573,P=0.002). Conclusion There is a significant correlation between single nucleotide polymorphism(SNP) at theGRB2rs8082005 andRXRArs3849222 loci and the risk of EOPE in pregnant women with gestational hypertension in Yanbian area.

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Objective To evaluate the anti-inflammatory effect of tissue inhibitor of metalloprotease(TIMP1) on a rat model of irreversible pulpitis. Methods An irreversible pulpitis model was established by pulp exposure on maxillary first molars of 8-10 weeks old male rats, and pulpotomy was performed at 24 h, 36 h and 48 h after pulp exposure, with TIMP-1 covering the pulp section(TIMP-1 group) and saline as control(NS group). Coronal obturation were performed with glass-ionomer cement. Histological and immunohistochemical analysis were performed at 24 and 48 h after administration. Results Quantitative histopathological analysis of HE-stained sections showed that the inflammation score(inflammatory cellular response of the root pulp, morphological changes in root pulp tissue and extent of root pulp necrosis) of the pulp were higher in the TIMP-1 group than those in the NS group(P<0.001 orP<0.01). Immunofluorescence staining(CD68-labeled) sections showed that the intensity of CD68 immunofluorescence was higher in the TIMP-1 group than that in the NS group(P<0.001). Conclusion TIMP-1 had no anti-inflammatory effect in a rat model of irreducible pulpitis and increased the degree of inflammation in the root pulp.

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Objective To evaluate the effect of 2.5%NaClO solution as irrigating agent on the healing of dental pulp after preservation treatment in rats with inflammatory pulp. Methods 48 maxillary molars of 24 8-week-old male SD rats were selected and pulpitis models were established by pulp cavity exposure for 24 hours. Then part of the crown pulp was removed and divided into two groups according to different irrigation methods: 2.5%NaClO flushing group(NaClO) and saline flushing group(NS). The pulp was covered with iRoot BP Plus and the cavity was sealed with glass ionomer. Four animals in the two groups were killed on the 1st, 7th and 28th day after operation, respectively. And the inflammatory reaction, tissue destruction and hard tissue formation of residual dental pulp tissue were observed and evaluated by histopathology. Results The pulp inflammatory reaction, tissue destruction and restorative dentin formation in 2.5%NaClO irrigation group and saline irrigation group were similar on the 1st, 7th and 28th day after operation, there was no significant difference between the two groups. Conclusion 2.5%NaClO used as a hemostatic before iRoot BP Plus pulp capping after partial pulpotomy has similar effect with normal saline on the inflammatory pulp model of rats, and does not affect the healing of pulp inflammation.

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Objective To explore differential expression of insulin like growth factor 1(IGF1) in oral squamous cell carcinoma(OSCC) tissues and cells, and to analyze its clinical significance of IGF1. Methods The protein expression of IGF1 was detected in 115 OSCC tissues(OSCC group) and 74 normal tissues(normal group) using immunohistochemical technique, and the relationship with the clinicopathological factors and prognosis of OSCC was analyzed. The IGF1 protein and mRNA levels in HOK and OSCC cell lines(CAL-27, TCA-8113, SCC-15 and SCC-25) were detected by Western blot and qRT-PCR.Results The high expression rate of IGF1 in OSCC group was 72.17%, which was significantly higher than that in normal group(2.70%)(P<0.001). The proportion under ROC curve(AUC) of OSCC diagnosed by IGF1 was 0.81, the sensitivity was 0.73, and the specificity was 0.82. The expression of IGF1 was related to the degree of differentiation, T stage and depth of invasion(P=0.03,P=0.02,P=0.02), but not to gender, age, N stage, TNM stage, smoking, alcohol consumption, or HPV infection(P>0.05). Kaplan-Meier and COX regression analysis showed that the high expression of IGF1, the degree of differentiation, the T stage and the depth of infiltration were the related factors affecting the prognosis of patients(P<0.01,P=0.04,P=0.03,P=0.04). COX multivariate indicated that high expression of IGF1 was an independent factor affecting the prognosis of patients(P=0.01). Western blot and qRT-PCR results indicated that the expression of IGF1 in OSCC cell lines was higher than that in HOK(P<0.05).Conclusion IGF1 can be a potential diagnostic and poor prognostic marker in oral squamous cell carcinoma.

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Abstract: Objective To study the preparation method of silane and bone morphogenetic protein-2(BMP-2) on the nano-porous titanium surface and evaluate the osteogenic ability of the prepared surface. Methods Silane and BMP-2 modified nano-porous titanium surfaces were constructed by alkali-heat treatment, silanization modification and BMP-2 grafting. Scanning electron microscopy, X-ray photoelectron spectroscopy, infrared spectroscopy and water contact angle tests were used to characterize the changes of surface morphology, elements, functional groups and hydrophilicity during the preparation process. Immunofluorescence was used to observe the BMP-2 grafted on the surface. Cell activity test and cell fluorescence staining were used to analyze the ability to promote osteoblast adhesion and proliferation, and alkaline phosphatase activity was used to evaluate the ability to promote cell osteogenesis. Results Material characterization results confirmed that silane and BMP-2 were successfully immobilized on the nano-porous titanium surface.Cell evaluation confirmed that the prepared surface not only significantly enhanced the adhesion and proliferation of osteoblasts, but also greatly enhanced the osteogenic ability of osteoblasts. Conclusion The constructed surface has better compatibility with osteoblasts and is expected to improve the osseointegration ability of titanium-based implants.