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Objective To investigate lim domain protein 4(LMO4) functions and mechanisms in regulating proliferation of skin squamous cells(A431), the shRNAs targeted to human LMO4 were coated by calcium phosphate nanoparticles(NP) and transfected into A431 cells to inhibit LMO4 expression. Methods Reverse transcription and quantitative polymerase chain reaction(RT-qPCR), immunohistochemistry analysis and Western blot were used to detect expression of the interest genes. The expression vectors with shRNA targeted to human LMO4(NP/sh-L) were coated by the calcium phosphate nanoparticles, and transfected into A431. The MTT assay was conducted to determine cell proliferation after transfected for 24, 36 and 48 h. Cells were stained with propidium iodide and examined cell cycles by using flow cytometry. Results LMO4 expressed at higher levels both in the skin squamous tissues and A431 cells. DNA-binding efficiency of NP/sh-L showed that the better combined ratio between calcium phosphate nanoparticles and DNA was 10 ∶1. There was no significant difference of transfection efficiency between the NP/sh-L and lipofection approaches. The MTT assay showed that silencing LMO4 inhibited proliferation of cells. RNAi-induced LMO4 inhibition exerted cell cycle arrest in G0/G1phase. Both CDK2 and cyclin E were downregulated in the A431 cells transfected with NP/sh-L comparing to the controls. However, LMO4 knockdown did not alter expression of CDK4 and cyclin D1. Conclusion The calcium phosphate nanoparticles could bind and transfer the foreign DNA into the targeted cells with high efficiency. Silencing LMO4 decreased expression of cyclin E and CDK2 resulted in inhibition of cell proliferation.

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Objective To investigate the effects of milk derived pentapeptide prolin-glycine-proline-isoleucine-proline(PGPIP) on chronic alcoholic liver injury in mice and its related molecular mechanism. Methods Forty C57BL/6 mice were randomly divided into control group, model group, glutathione(GSH) group, PGPIPN group. The mice model of chronic alcoholic fatty liver was established by 10 d ad libitum oral feeding with the Lieber-DeCarli(LD) ethanol liquid diet plus one binge. Drug intervention was given at the same time. Based on the reported RNA sequencing data in gene expression omnibus(GEO) database, the differential expression of liver endoplasmic reticulum stress-related genes was analyzed by cluster heat map. Liver hematoxylin-eosin(HE) staining was used to analyze the pathological effects of each treatment group on alcoholic liver injury in mice. Oiled Red O staining was used to analyze the effects of each treatment group on the accumulation of liver lipid droplets in mice. Transmission electron microscopy was used to analyze the effect of lipid droplet accumulation caused by alcohol metabolism on the microstructure of hepatocytes and verify the effect of PGPIP. Endoplasmic reticulum stress-related signal transduction protein expression was detected by Western blot. Results The pathological examination of PGPIP group was similar to that of Control group, and the liver injury of mice was significantly reduced. The accumulation of lipid droplets in the liver of mice in the model group was manifested as mixed lipid droplets of different sizes, and PGPIP treatment significantly reduced the accumulation of liver lipid droplets induced by alcohol. PGPIP had significant effects on the PERK-eIF2α-ATF4 pathway and the expression of transcriptional activator 6(ATF6), Cleaved Caspase 3 protein. Conclusion Pentapeptide PGPIP can alleviate chronic alcoholic fatty liver and liver injury in mice. The mechanism may be attributed to reducing the accumulation of lipid droplets in hepatocytes, endoplasmic reticulum stress, and hepatocyte apoptosis.

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Objective To investigate the cross-sectional associations of serum interleukin(IL)-18 with cartilage volume, cartilage defects, bone marrow lesions(BML) and biomarkers of cartilage degradation in patients with knee osteoarthritis(OA), and to provide new ideas and new methods for clinical diagnosis and treatment. Methods The study included 151 patients with knee OA, a general questionnaire survey was conducted, and the knee structural was photographed by magnetic resonance imaging(MRI). The cartilage volume was measured by OsiriX software in 3D-FLASH sequence, and cartilage defect and BML were determined in T2-weighted sequence. Serum IL-18 and matrix metalloproteinase(MMP)-3, 13 levels were measured by enzyme-linked immunosorbent assay(ELISA). SPSS software was used for statistical analysis.Results In multivariable analyses, serum IL-18 level was negatively associated with total cartilage volume(β=-0. 24, 95%CI:-0. 13-0. 03) and the association was consistent at divided part of joint(femorotibial joint and the patella femoral joint, allP<0. 05). Serum IL-18level was positively associated with cartilage defect and BML at media femorotibial area(allP<0. 01). Serum IL-18 level was positively associated with MMP-3(β= 0. 31, 95%CI:0. 001-0. 010) and MMP-13(β= 0. 86, 95%CI:0. 08-0. 10, allP<0. 01).Conclusion Serum IL-18 level is negatively associated with cartilage volume and positively associated with cartilage defect, BML, MMP-3 and MMP-13, suggesting IL-18 may play a significant role in the progression of cartilage degradation in patients with knee OA, the therapy of targeting serum IL-18 may reduce the injury of article cartilage in patients with knee OA and delay the progression of disease.

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Objective To explore a non-invasive assay for screening osteosarcoma patients with positive programmed death receptor-1(PD-1)/programmed death receptor ligand-1(PD-L1) signaling pathway expression. Methods The subcutaneous tumor bearing mouse model of human Osteosarcoma cells(OS-732) was established by the method of tumor formation, and toxicity test was performed to verify the toxicity of PD-L1 antibody to mouse organs.124I-anti-PD-L1 monoclonal antibody molecular probe was further synthesized, and OS-732 rats were injected with 18.5 MBq124I-anti-PD-L1 probe in tail vein to start OS-732 osteosarcoma Micro Positron Emission Tomography/Computed Tomography(Micro-PET/CT) imaging. Results OS-732 osteosarcoma model was successfully constructed, immunohistochemistry confirmed the presence of PD-L1 expression in OS-732 osteosarcoma, and Micro-PET/CT imaging was successfully performed at different time points(2, 24 and 48 h after probe injection) to achieve non-invasive real-time observation of PD-L1 in OS-732 osteosarcoma. Conclusion In this study, the124I-anti-PD-L1 monoclonal antibody molecular probe was constructedin vitro, and Micro PET/CT imaging verified that the probe successfully targeted the PD-L1 receptor of OS-732 Osteosarcoma, and showed clear immune imaging, indicating that it is hopeful to achieve non-invasive screening of Osteosarcoma patients with PD-L1 positive expression in clinical practice, and this technology might benefit the majority of Osteosarcoma patients.

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Objective To observe the expression of glial fibrillary acidic protein(GFAP) in the brain tissue of rats with traumatic brain injury(TBI) and the differential changes between brain regions. Methods The TBI model of rats was prepared by the modified Feeney free fall method, and it was randomly divided into the sham-operated group(Sham group) and the model group(TBI group). Immunohistochemical staining, Western blot and RT-qPCR were used to observe the morphological changes of activated astrocytes and the expression of GFAP in different brain regions after TBI. Results Immunohistochemical staining showed that astrocytes were activated in allbrain regions on the right side of TBI, and the only brain regions that spilled over to the contralateral side(left) were cortical region 1 and midbrain regions. RT-qPCR results showed that GFAP mRNA was highly expressed in all brain regions on the right side of TBI, affecting the contralateral cortex and midbrain regions. Western blot results showed that GFAP protein was highly expressed in all brain regions on the right side of TBI, and it spread to the contralateral midbrain regions. Conclusion TBI can cause the activation of astrocytes and increase GFAP expression in the right brain regions, while also affecting the contralateral(left) cortex 1 and midbrain regions.

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Objective To explore the expression of the μ-opioid receptor(MOR) and the effects of intracellular vesicle transport on the MOR expression during endomorphin 2(EM2) analgesia. Methods Adult male SD rats were randomly divided into 3 groups: control(naive) group, neuropathic pain group and drug group. Spared nerve injury(SNI) induced neuropathic pain rats were employed as the pain model. The drug group rats were the SNI pain ones intrathecally injected with EM2. The methods of immunofluorescence single staining and Western blot were used to detect the expression of MOR total protein, phosphorylated protein and Rab7 protein. Immunofluorescence double staining was used to detect the expression of MOR/Rab7 and MOR/LAMP1 co-labeled immunoreactivity. Results Compared with the control group, the expression of total MOR protein and phosphorylated protein in the dorsal root ganglion(DRG) of the SNI pain rats decreased(P<0.05), and the expression of Rab7 significantly increased(P<0.05). The expression of MOR/Rab7 co-labeled immunoreactivity in Rab7 and MOR immunoreactive(-ir) products and MOR/LAMP1 co-labeled immunoreactivity in MOR and LAMP1-ir products both increased(P<0.05). Multiple intrathecal injection of EM2 significantly increased paw withdrawal threshold in the SNI neuropathic pain rats(P<0.01), the expression of MOR protein and phosphorylated protein in DRG was increased(P<0.05), while the expression of Rab7 decreased(P<0.05). Compared with the control group, the expression of MOR/Rab7 positive products in Rab7 and MOR positive ones decreased, and the expression of MOR/LAMP1 positive products in LAMP1 and MOR positive markers decreased(P<0.05). Conclusion In the process of analgesia, EM2 inhibits the expression of Rab7 in the DRG of SNI neuropathic pain rats, reduces the transport of MOR to lysosomes, and promotes the re-sensitization of MOR.

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Objective To observe the effects of electroacupuncture and bifidobacterium on the survival state, the level of intestinal signaling substances and lipid metabolism levels in rats with simple obesity, and to compare their efficacy. Methods Sixty 4-week-old SPF SD rats with the body weight ranging from 40 to 60 g were purchased, 10(half male and half female) were given ordinary diet as blank group, and 50(half male and half female) were given high fat diet to prepare simple obese rat model. After 8 weeks, 30 obese rats successfully established in high fat group were randomly divided into obesity model group, electroacupuncture group and bifidus group, with 10 rats in each group(half male and half female). The treatment group was treated with electroacupuncture and bifidobacterium solution for 4 weeks, respectively, and the body mass of the rats was measured at a fixed time during the experiment. After the treatment cycle, the level of enterochromaffin cell(ECC) in colon was detected by silver staining, the levels of 5-hydroxytryptamine(5-HT), histamine(HIS) in obese rats were detected by ELISA, the levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein(LDL) and high density lipoprotein(HDL)were detected by biochemical method. Results After intervention, compared with model group, body weight, colon and hypothalamus 5-HT, HIS, colon ECC, serum TC, TG and LDL levels decreased in electroacupuncture group and bifidobacterium group, with statistical significance(P<0.05), although the serum HDL levels were increased, the difference was not statistically significant. And the indexes of the electroacupuncture group were more similar to those of the blank group than those of the bifidobacterium group. Conclusion The effect of electrotherapy on simple obesity rats is better than bifidobacterium. Electroacupuncture may stimulate the hypothalamus to reduce food intake, accelerate lipid metabolism and slow down obesity by adjusting the intestinal signaling substances in rats with simple obesity.

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Objective To investigate the effect and mechanism of miR-145-3p on mitophagy in 1-methyl-4-phenylpyridiniumion(MPP+)-induced Parkinson′s disease(PD) cell model. Methods Human neuroblastoma cells(SH-SY5Y) were divided into control group, model group, mimics group, calmodulin-dependent protein kinase kinaseβ(CaMkkβ) inhibitor(STO-609) group, mimics+STO-609 group, cyclic adenosine monophosphate response element-binding protein(CREB) inhibitor(KG-501) group, mimics+KG-501 group and STO-609+KG-501 group. Cell apoptosis was detected by flow cytometry, autophagosome structure was observed by transmission electron microscopy, and apoptosis, autophagy and CaMkkβ/adenylate activated protein kinase(AMPK)/CREB pathway related protein expression were detected by Western blot. Results Compared with control group, the apoptosis rate, Bcl-2-associated X protein(Bax), cysteine proteinase-3(Caspase-3) and microtubule-associated protein light chain 3-I(LC3-Ⅰ) protein expression levels in model group increased(P<0.01), and the autophagosome structure decreased. The protein levels of B cell lymphoma-2(Bcl-2), autophagy gene(Beclin-1), microtubule-associated protein light chain 3-Ⅱ(LC3-Ⅱ), phosphorylated calmodulin-dependent protein kinase kinaseβ(p-CaMkkβ), phosphorylated cadenylate activated protein kinase(p-AMPK), and phosphorylated cyclic adenosine monophosphate response element-binding protein(p-CREB) decreased(P<0.01). Compared with model group, the apoptosis rate, Bax, Caspase-3 and LC3-Ⅰ protein expression levels in mimics group decreased(P<0.05), and the autophagosome structure increased. The protein levels of Bcl-2, Beclin-1, LC3-Ⅱ, p-CaMkkβ, p-AMPK, p-CREB increased(P<0.05). The trend of STO-609 group and KG-501 group was the same and opposite to mimics group. Compared with mimics group, the apoptosis rate, Bax, Caspase-3 and LC3-Ⅰ protein expression levels in the mimics+STO-609 group and the mimics+KG-501 group increased(P<0.01), and the autophagosome structure decreased. The protein levels of Bcl-2, Beclin-1, LC3-Ⅱ, p-CaMkkβ, p-AMPK, p-CREB protein levels decreased(P<0.01). Compared with STO-609 group, the apoptosis rate, Bax, Caspase-3 and LC3-Ⅰ protein expression levels of STO-609+KG-501 group increased(P<0.01), and the autophagosome structure decreased. The protein levels of Bcl-2, Beclin-1, LC3-Ⅱ, p-CaMkkβ, p-AMPK and p-CREB decreased(P<0.05). Conclusion miR-145-3p can inhibit the apoptosis of MPP+-induced PD cell model and promote mitophagy, and its mechanism may be related to the activation of the CaMkkβ/AMPK/CREB pathway.

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Objective To study the association between single nucleotide polymorphism(SNP) in transcriptional coactivator containing PDZ binding motif(TAZ) gene and the risk of non-cardia gastric cancer. Methods Among 460 patients with non-cardia gastric cancer and 470 normal controls, TAZ rs16861985, rs1055153, rs6783790, rs12490239 were genotyped by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Unconditional Logistic regression was used to evaluate the association of studied SNPs with the risk of non-cardia gastric cancer. Results TAZ rs16861985 was associated with the risk of non-cardia gastric cancer, compared with the CC genotype carriers, the carriers of GC genotype had an increased risk of non-cardia gastric cancer(OR=1.547, 95%CI: 1.145-2.089). TAZ rs1055153 was associated with the risk of non-cardia gastric cancer, compared with the GG genotype carriers, the carriers of GT+TT genotypes had a decreased risk of non-cardia gastric cancer(OR=0.570, 95%CI: 0.400-0.814). TAZ rs6783790 and rs12490239 were not associated with the risk of non-cardia gastric cancer. Among the haplotypes constructed by TAZ rs16861985, rs1055153, rs6783790 and rs12490239, C-G-A-A haplotype had a decreased risk of non-cardia gastric cancer(OR=0.761, 95%CI: 0.612-0.945), while G-G-A-G haplotype had an increased risk of non-cardia gastric cancer(OR=1.894, 95%CI: 1.314-2.730). The fourth-order interaction of TAZ rs16861985, rs1055153, rs6783790 and rs12490239 was associated with non-cardia gastric cancer risk(P<0.05). Conclusion TAZ rs16861985 and rs1055153 may play a role in the risk of non-cardia gastric cancer. The C-G-A-A haplotype constructed by TAZ rs16861985, rs1055153, rs6783790 and rs12490239 can reduce the risk of non-cardia gastric cancer, and G-G-A-G haplotype can increase the risk of non-cardia gastric cancer. The interaction of TAZ rs16861985, rs1055153, rs6783790 and rs12490239 has a synergistic effect innon-cardia gastric cancer.

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Objective To investigate the effects of aerobic exercise intervention at low temperature on renal metabolism and short-term prognosis in diabetic nephropathy. Methods (1) 32 mice with diabetic nephropathy were randomly divided into a model group, a low-temperature group, an exercise group, and a low-temperature exercise group, and another 8 mice were selected as members of the normal group. The exercise group and the low-temperature exercise group were given an 8-week running table exercise intervention at room temperature and low temperature, respectively, while the low-temperature group was only given the low temperature intervention. Fasting blood glucose(FBG), serum creatinine(Scr), blood urea nitrogen(BUN), and 24 h urine protein levels were measured by a biochemical analyzer; renal lesions were observed by the hematoxylin-eosin(HE) method; and nuclear factor E2-related factor 2(NRF2) and heme oxygenase 1(HO-1) levels in renal tissues were measured by Western blot.(2) 194 patients with diabetic nephropathy were prospectively enrolled and divided into the exercise group(1 time/week,n=35), the exercise group(≥2 times/week,n=47), and the control group(0 time/week,n=112) according to the frequency of aerobic exercise intervention at low temperature. Scr, BUN, uric acid(UA), urinary albumin/creatinine ratio(UACR), and estimated glomerular filtration rate(eGFR) levels were compared in the 3 groups. Kaplan-Meier and COX regression were performed to analyze the relationship between aerobic exercise interventions at low temperature and short-term prognosis. Results (1) Compared with the normal group, the levels of FBG, Scr, BUN, 24 h urine protein, and the kidney injury score in the model group increased, while the levels of NRF2 and HO-1 in the model group decreased(P<0.05). Compared with the model group, the levels of FBG, Scr, BUN, 24 h urine protein, and the kidney injury score in the exercise group and the low-temperature exercise group decreased, while the levels of NRF2 and HO-1 in the exercise group and the low-temperature exercise group increased(P<0.05). Compared with the low-temperature group and the exercise group, the levels of FBG, Scr, BUN, 24 h urine protein, and the kidney injury score in the low-temperature exercise group decreased, while the levels of NRF2 and HO-1 in the low-temperature exercise group increased(P<0.05).(2) After the aerobic exercise intervention at low temperature, the levels of Scr, BUN, and UACR in the exercise group(1 time/week) and the exercise group(≥2 times/week) were lower than those in the control group, and eGFR were higher than that in the control group(P<0.05). The cumulative incidence of poor prognosis in the exercise group(≥2 times/week) was 2.13%(1/47), which was lower than in the exercise group[1 time/week, 11.43%(4/35)] and the control group[16.07%(18/112),P<0.05]. COX regression analysis showed that exercise intervention ≥2 times/week(HR=0.123, 95%CI: 0.016-0.925) reduced the risk of poor short-term prognosis in patients with diabetic nephropathy compared with no intervention. Conclusion Aerobic exercise intervention at low temperature can correct renal metabolic abnormalities in diabetic nephropathy, protect renal tissues, and reduce the risk of poor short-term prognosis by a mechanism that may be related to upregulation of NRF2/HO-1 axis expression.

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Objective To construct folate modified D-α-tocopheryl polyethylene glycol 1000 succinate(TPGS) nanomaterials(FA-TPGS), which can stably load and effectively release siRNA, and to observe the effects of nanoparticles on the cytotoxicity and spliced X-box binding protein 1(XBP1s)of mouse leukemia cells of monocyte macrophage(RAW264.7). Methods Mixed FA-TPGS and rhodamine B(RhB) labeled XBP1 siRNA solution in a proportion of 5 ∶1 and obtained the nano-complex coated with XBP1 siRNA(FT@XBP1). FT@XBP1 nanocarriers were characterized by transmission electron microscope, dynamic light scattering, ultraviolet visible spectrum analysis and/or fluorescence analysis. And the release of siRNA from FA-TPGS nano-carriers was calculated simultaneously. The cell cytotoxicity of FT@XBP1 nanomaterials were detected by scanning electron microscopy(SEM), CCK-8 and flow cytometry. And the inhibited effect of XBP1s of RAW264.7 cells was checked by Western blot. Results FA modified TPGS could effectively bind XBP1 siRNA. And the average particle size of FT@XBP1 nanocarriers were(200±20)nm. The relative release assays showed that acidic environments(pH 5.0)was beneficial for siRNA to release from FT@XBP1. Both CCK-8 and apoptosis assay showed that the effects of FT@XBP1 on the proliferation and apoptosis of RAW264.7 cells were relatively small, and FT@XBP1 could significantly inhibit the expression of XBP1s protein in RAW264.7(P<0.001). Conclusion TPGS nanoparticles modified with folic acid can effectively encapsulate XBP1 siRNA and inhibit XBP1s expression of RAW264.7 cells with good cellular compatibility.

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Objective To investigate the effect of adeno-associated virus(AAV) delivery of short hairpin RNA(shRNA) against the Ptger3 gene in the lateral parabrachial nucleus(LPB) on the fever induced by microinjection of prostaglandin E2(PGE2) into the LPB and the intraperitoneal injection of lipopolysaccharide(LPS). Methods AAV2-shRNA-Ptger3(EP3)-EGFP(shRNA-EP3) and AAV2-CMV-EGFP(shRNA-control) viruses were constructed and transfected the rat LPB by stereotaxic injection. Four weeks later, the transfection efficiency of AAV viruses was observed by fluorescence microscopy, and the knockdown efficiency was determined by real-time PCR of EP3 receptor mRNA on the LPB. The effects of microinjection of saline or PGE2in the LPB or intraperitoneal injection of LPS on body temperature(Tcore) and energy expenditure(EE) of shRNA-control group and shRNA-EP3 group were monitored using an animal monitoring system with temperature telemetry. Results AAV virus transfection of shRNA-EP3 group and shRNA-control group was mainly found in the LPB. The EP3 receptor mRNA level in LPB of shRNA-EP3 group was reduced compared with that of shRNA-control group(P<0.05). There was no significant difference in basal body temperature between shRNA-control group and shRNA-EP3 group. Tcoreand EE were briefly and slightly increased after microinjection of saline in the LPB, but there was no significant difference between the two groups. Compared with the shRNA-control group, the febrile response induced by LPB PGE2was attenuated in the shRNA-EP3 group(P<0.05). Furthermore, the knockdown of EP3 receptor of LPB also attenuated the LPS-induced fever, and the Tcore5.5 h post-LPS in the shRNA-EP3 rats increased compared with the baseline(P<0.05), which was lower than that in the shRNA-control rats(P<0.01). Conclusion EP3 receptor knockdown in LPB attenuates the febrile response induced by microinjection of PGE2in the LPB and intraperitoneal injection of LPS, suggesting that EP3 receptors of LPB mediate the pyrogenic action of LPB PGE2and partly participate in LPS-induced fever.

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Objective To analyze the expression and clinical significance of circ-RACGAP1 in bladder cancer tissues, and to explore the influence and mechanism of circ-RACGAP1 on the malignant biological behavior of bladder cancer cells. Methods The expression of circ-RACGAP1 in bladder cancer tissues was explored through the TCGA database, and the relationship between the expression of circ-RACGAP1 and the clinicopathological features of bladder cancer patients was analyzed. The expression of circ-RACGAP1 in cell lines 5637, T24, J82, RT-4 and UM-UC-3 was analyzed by quantitative real-time PCR(qPCR). The circ-RACGAP1 knockdown plasmid was transfected into T24 cells by lipofection technology. Colony formation assay, scratch assay and Transwell assay were used to analyze the effects of knocking down circ-RACGAP1 on the proliferation, migration and invasion of T24 cells, respectively. The targeted binding between circ-RACGAP1 and miR-4324 was verified using deepBase, Circbank, CircInteractome, circRNABase databases and a fluorescent reporter system. The effect of knocking down circ-RACGAP1 on the expression of miR-4324 in T24 cells was detected by qPCR. Western blot was used to detect the effect of knocking down circ-RACGAP1 on the expression of recombinant rac-GTPase activating protein 1(RACGAP1) protein and phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT) signaling pathway proteins in T24 cells. Results circ-RACGAP1 was highly expressed in bladder cancer tissues(P<0.01), and its expression increased with the clinical stage of the patients(P<0.01). The expression of circ-RACGAP1 in bladder cancer cell lines was significantly higher than that in normal human bladder epithelial cells(all P<0.01). Compared with the control group, the proliferation, migration and invasion abilities of T24 cells in the sh-circ-RAC-GAP1 group significantly decreased(allP<0. 01). circ-RACGAP1 could target and inhibit the expression of miR-4324(P<0. 01). Compared with the control group, the expression level of RACGAP1 protein in T24 cells in the sh-circ-RACGAP1 group decreased(P<0. 01), and the expression levels of PI3K/AKT signaling pathway proteins phosphatidylinositol-3-kinase(p-PI3K), phosphorylated protein kinase B(p-AKT), nuclear factor kappa B(NF-κB) decreased(allP<0. 01).Conclusion circ-RACGAP1 is highly expressed in bladder cancer tissues and cell lines, knocking down circ-RACGAP1 can inhibit the malignant biological behavior of T24 cells, and circ-RACGAP1 plays a role by inhibiting the expression of miR-4324 and activating the PI3K/AKT signaling pathway.

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Objective To explore the nucleotide variation and protein amino acid changes of E4 and L2 genes ofHuman Papillomavirus16(HPV16), and to analyze the evolutionary characteristics of HPV16 virus. Methods 40 HPV16 infection-positive cervical exfoliated cells samples and tissue cell samples were collected from hospital, viral DNA was extracted, Sanger sequencing perform in cervical exfoliated cells DNA and high-throughput sequencing technology sequenced in cervical tissues DNA for E4 and L2 genes of HPV16, HPV16 E4 and L2 gene phylogenetic evolution trees were constructed, and variation of HPV16 E4 and L2 genes were analyzed. Results There were 72 HPV16 E4 variant samples with nucleotide variants(4 missense mutations and 7 synonymous mutations) at 10 sites, HPV16 L2 gene variants in 74 samples, and nucleotide variants(23 missense mutations and 18 synonymous mutations) at 40 sites. The variation frequency of T4177C, A4288C and A4654C in cervical cancer was significantly higher than that in non-cervical cancer, and the difference was statistically significant(P<0.05). Conclusion (1) The main HPV16 virus strains in Xinjiang are European strains, and a few are Asian strains.(2) The mutation frequency of T4177C, A4288C and A4654C in HPV16 L2 gene is higher than that in non-cervical cancer, and G4181A is related to the Asian strain.

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Objective To explore the regulatory effects of ResolvinD1(RVD1) on autophagy and apoptosis of cardiomyocytes in aging rats, and the effects of RVD1 on mitochondria-associated endoplasmic reticulum membranes(MAMs) of myocardial tissue. Methods Thirty 18-month-old SD rats were randomly divided into model group, low-dose RVD1 group and high-dose RVD1 group, with 10 rats in each group, and another 10 6-month-old SD rats were selected as control group. The low-dose RVD1 group and high-dose RVD1 group were injected with 50 ng/ml and 100 ng/ml RVD1 through the tail vein, respectively, control group and model group were injected with the same amount of phosphate buffered saline(PBS) for 21 days. The β-galactosidase activity of rat myocardial tissue was determined by p-nitrophenol method, the histopathological changes of rat myocardial tissue were detected by hematoxylin-eosin(HE) staining, and the collagen deposition in rat myocardial tissue was observed by Masson staining, apoptosis of rat myocardial cells was detected by dUTP in situ end labeling mediated by terminal deoxynucleotide transferase(TUNEL), the expression of autophagy marker microtubule-associated protein 3(LC3) was detected by immunohistochemical S-P method, the ratio of LC3-Ⅱ/LC3-Ⅰ and the expression of Beclin-1, p62 in rat myocardial tissue were determined by Western blot, the structure of MAMs in rat myocardial tissue was observed by transmission electron microscopy, the expression levels of glucose-regulatory protein 75(GRP75), volt-dependent anion channel 1(VDAC1) and mitochondrial fusion protein 2(Mfn2) in rat myocardial tissue were determined by Western blot. Results Compared with model group, β-galactosidase activity of myocardial tissue decreased in low-dose RVD1 group and high-dose RVD1 group(P<0.05), myocardial fiber breakage and myocardial cell damage were significantly alleviated, collagen fiber deposition was significantly reduced, and the proportion of TUNEL positive cells in myocardial tissue decreased(P<0.05), the positive rate of LC3 protein increased(P<0.05), the ratio of LC3-Ⅱ/LC3-Ⅰ increased, the relative expression level of Beclin-1 protein was up-regulated and the relative expression level of p62 protein was down-regulated(P<0.05), the structure of MAMs was tighter, and the percentage of MAMs in mitochondrial circumference also increased(P<0.05), the relative protein expressions of GRP75, VDAC1 and Mfn2 were up-regulated(P<0.05). Compared with low-dose RVD1 group, β-galactosidase activity in high-dose RVD1 group further decreased(P<0.05), no obvious damage was observed in myocardial tissue, collagen fiber deposition was further decreased, the proportion of TUNEL positive cells in myocardial tissue decreased(P<0.05), and the positive rate of LC3 protein increased(P<0.05), LC3-Ⅱ/LC3-Ⅰ ratio increased, Beclin-1 relative expression level was up-regulated and p62 relative expression level was down-regulated(P<0.05),the structure of MAMs was more compact, and their percentage in mitochondrial circumference further increased(P<0.05), the relative expressions of GRP75, VDAC1 and Mfn2 were further up-regulated(P<0.05). Conclusion RVD1 can activate autophagy, alleviate myocardial apoptosis and collagen fiber deposition, and promote mitochondria-associated endoplasmic reticulum membranes in aging rats.

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Objective To explore the effects of long non-coding RNA LINC00092 on the proliferation, migration and invasion of glioma. Methods Using the cancer genome atlas(TCGA) and genotype-tissue expression(GTEx) databases, this study analyzed the expression of LINC00092 in pan-carcinoma and its effect on the prognosis of glioma. In addition, LINC00092 overexpression plasmid was constructed to detect the effects of LINC00092 on proliferation, migration, invasion and apoptosis of glioma cells by cell function experiments, including CCK-8 assay, Transwell assay and flow cytometry. Finally, qRT-PCR and Western blot were used to detect the effect of overexpression of LINC00092 on the expression level of IGF2BP1. Results The analysis of public databases revealed a widespread downregulation of LINC00092 in tumors, and its association with the development of glioblastoma multiforme(GBM) and low-grade glioma(LGG).In vitroexperiments demonstrated that overexpression of LINC00092 significantly reduced the proliferation, migration and invasion of glioma cells, while promoting apoptosis. Moreover, overexpression of LINC00092 led to a decrease in the expression levels of IGF2BP1. Conclusion LINC00092 may inhibit glioma proliferation, migration and invasion by targeting IGF2BP1, and promote glioma cell apoptosis.

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Objective To used Toll-like receptor 4(TLR4)inhibitor(TAK-242) investigate the role of TLR4 in the early stage of acetaminophen(APAP)-induced acute liver injury in mice. Methods Fifty-eight male institute of cancer research(ICR) mice were randomly divided into control group(normal control group, solvent control group, inhibitor control group) and experimental group(APAP 4-hour group, APAP+TAK-242 4-hour group, APAP 12-hour group, APAP+TAK-242 12-hour group). Mice in the experimental group were given a single dose of APAP(300 mg/kg) and TAK-242(3 mg/kg) were given intraperitoneal injection 3-hour before APAP injection. The serum alanine aminotransferase(ALT) level and liver index of mice in each group were compared; HE staining showed pathological changes of liver tissue; the level of high mobility group box-1(HMGB1) was determined by immunohistochemistry; the levels of TLR4, HMGB1 and nuclear factor kappa B(NF-κB)protein were detected by Western blot; the levels of monocyte chemoattractant protein-1(MCP-1), interleukin(IL)-1β, tumor necrosis factor-α(TNF-α)and IL-6 genes were determined by real-time fluorescence quantitative PCR(RT-qPCR). The content of IL-6 in liver tissue was determined by enzyme linked immunosorbent assay(ELISA). Results HE staining showed liver tissue of mice obvious swelling and congestion of in APAP group at 4-hour and typical lobular central necrosis in APAP group at 12-hour; the degree of liver necrosis in APAP+TAK-242 groups at 4-hour and 12-hour was less than that in APAP group at the same time point. Compared with the normal control group, serum ALT level, liver index, TLR4, HMGB1, NF-κB protein content, MCP-1, IL-1β, TNF-α, IL-6 gene levels and IL-6 content in liver tissue of APAP 4-hour group increased;serum ALT level, liver index, HMGB1protein content and IL-6 content in liver tissue increased in APAP 12-hour group. Compared with APAP 4-hour group, serum ALT level, liver index, TLR4, HMGB1, NF-κB protein content, MCP-1, IL-1β, TNF-α, IL-6 gene level and IL-6 content in liver tissue decreased in APAP + TAK-242 4-hour group. Compared with APAP 12-hour group, the levels of TLR4, HMGB1 protein and MCP-1, IL-1β, TNF-α genes in APAP + TAK-242 12-hour group decreased.Conclusion Inhibition of TLR4 may inhibit TLR4/HMGB1 pathway to reduce the inflammatory response in the early stage of acetaminophen-induced acute liver injury in mice.

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Objective To investigate the effects of melatonin(MEL) on the proliferation of hepatic stellate cells(HSCs) induced by platelet-derived growth factor(PDGF-BB) and explore its correlation with the regulation of autophagy levels. Methods The HSC-T6 cells were divided into the following groups: control group, model group and MEL(low, medium and high) treatment groups. After 24 hours culture, the cells adhered to the wall and were changed into serum-free DMEM medium to synchronize the cells to the G0stage. After 24 hours culture, all groups were given with PDGF-BB(10 ng/ml) excepted the control group. Besides, melatonin of different concentrations(1 nmol/L, 1 μmol/L and 0.1 mmol/L) were added immediately in three treated groups. After incubated for 48 hours, the effect of MEL on the proliferation of hepatic stellate cells activated by PDGF-BB was detected by CCK-8 method. The protein expression levels of LC3b and α-SMA in hepatic stellate cells were determined by Western blot. The expression levels of LC3b mRNA and α-SMA mRNA in hepatic stellate cells were determined by qRT-PCR. The ultrastructure of HSCs was observed by transmission electron microscopy to understand the autophagy level. Results Compared with control group, PDGF-BB could induce the proliferation of HSCs(P<0.01). Compared with model group, MEL inhibited the proliferation of HSCs activated by PDGF-BB(P<0.01). Compared with the control group, LC3b and α-SMA protein expressions significantly increased in the model group(allP<0.05), and LC3b mRNA and α-SMA mRNA expressions significantly increased in the model group(P<0.05,P<0.01). Compared with the model group, MEL could inhibit such effects(LC3b:P<0.05,P<0.01; α-SMA:P<0.01). Transmission electron microscopy(TEM) showed that compared with the control group, autopolysosome significantly increased in the model group(P<0.05). Compared with model group, autopolysosome significantly decreased in MEL treatment group(P<0.01). Conclusion The up-regulation of autophagy level can promote the proliferation of hepatic stellate cells and the inhibition of hepatic stellate cell proliferation by MEL may be related to the down-regulation of autophagy level.

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Objective To investigate the effects of hypoxia, high glucose single factor and hypoxia high glucose compound factors on the pyroptosis of rat glomerular podocytes. Methods Rat glomerular podocytes were culturedin vitroand randomly divided into control group, high glucose group, hypoxia group and hypoxia high glucose group, EdU method was used to detect the cell proliferation, transmission electron microscope was used to observe the morphology and size changes of nucleus and mitochondria, and Western blot was used to detect pyroptosis related proteins nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3), Cysteinyl aspartate specific proteinase-1(Caspase-1), gasdermin(GSDMD) and inflammatory factor pro-interleukin-1β(Pro-IL-1β),interleukin(IL)-1β,IL-18. The effect of hypoxia and high glucose on the pyroptosis of rat glomerular podocytes was analyzed. Results EdU results showed that hypoxia and high glucose inhibited the proliferation ability of rat glomerular podocytes(P<0.05). The results of transmission electron microscopy suggested that hypoxia and high glucose promoted the occurrence of pyroptosis of rat glomerular podocytes. Western blot showed that hypoxia and high glucose promoted pyroptosis of rat glomerular podocytes, and increased the expression of pyroptosis related proteins NLRP3, Caspase-1 and GSDMD, among which the expression of pyroptosis protein increased most significantly in hypoxia and high glucose group(P<0.05). At the same time, it also increased the expression of pro-inflammatory factor Pro-IL-1β, IL-1β and IL-18(P<0.05). Conclusion hypoxia and high glucose can induce pyroptosis of rat glomerular podocytes, one of the mechanisms may be through affecting NLRP3-Caspase-1-GSDMD and its downstream inflammatory factors.

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Objective To investigate the effect of SHP2 protein phase separation induced by activation mutation on cell proliferation and its mechanism through construct a mouse model of SHP2E76Kmutation. Methods Hybrid PTPN11E76K-NEO/+C57BL/6J mouse were hybridizedwith Mx1-cre tool mice to obtain the required Mx1-cre; Ptpn11+/+and Mx1-cre; Ptpn11E76K/+. The later genotype mice were injected with pI-pC to induce the expression of Cre enzyme and mutate Ptpn11E76K in bone marrow mesenchymal stem cells(MSC).The Mx1-cre; Ptpn11+/+and Mx1-cre; Ptpn11E76K/+genotype mice′s cells were isolated and culturedin vitroand identified as MSCs by immunofluorescence staining.With Ptpn11+/+MSC as the control group and Ptpn11E76K/+MSC as the experimental group, the two kinds of cells were divided into 6 groups by adding drugs: Ptpn11+/+group; Ptpn11+/++SHP099 group; Ptpn11+/++ET070 group; Ptpn11E76K/+group; Ptpn11E76K/++SHP099 group; Ptpn11E76K/++ET070 group. The differences of SHP2 protein phase separation in the six groups were observed by immunofluorescencestaining, and the differences of SHP2 protein expression were detectedby Western blot. CCK-8 was used to observe the changes of cell proliferation after phase separation was affected. Western blot was used to detect the expression levels of ERK/p-ERK,AMPK/p-AMPK, mTOR/p-mTOR and other molecules between the six groups. Results Genotypes Mx1-cre; Ptpn11E76K/+and Mx1-cre; Ptpn11+/+mice were obtained by genotyping, and the primary MSCs were isolated.Compared with Ptpn11+/+group, SHP2 proteins in the Ptpn11E76K/+group produced more phase separation condensates, and compared with Ptpn11E76K/+group, the SHP2 proteins in the Ptpn11E76K/++SHP099 and Ptpn11E76K/++ET070 groups produced less phase separation condensates.No difference in SHP2 protein expression levels between groups was detected by Western blot. Compared with Ptpn11+/+group, the proliferation ability of MSC in Ptpn11+/++SHP099 and Ptpn11+/++ET070 groups decreased, the expression of p-ERK and p-mTOR decreased, and the expression of p-AMPK protein increased. Compared with Ptpn11E76K/+group, the proliferation ability of MSC in Ptpn11E76K/++SHP099 and Ptpn11E76K/++ET070 groups decreased, the expression of p-ERK and p-mTOR decreased, and the expression of p-AMPK protein increased. Conclusion SHP2 phase isolation is involved in the alteration of proliferative capacity of SHP2E76K-activated MSCs by stimulating the expression of AMPK-mTOR signaling pathway.

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Objective To investigate the expression of class A scavenger receptor 1(MSR1) in the lungs of silicosis mice and its role in inflammation and lipid metabolism mediated by mouse mononuclear macrophages(RAW264.7). Methods 24 C57BL/6 male mice were randomly divided into control group, exposed 7 d group, exposed 14 d group, exposed 28 d group, with 6 mice in each group. RAW264.7 cells were divided into control group, siRNA-MSR1 group, SiO2group and siRNA-MSR1+SiO2stimulation group. The pathological changes of lung tissue in mice were observed by HE and VG staining. Lipid accumulation was observed under oil red O staining microscope. Immunohistochemical staining(IHC) was used to detect the expression and localization of MSR1. The expression of MSR1, tumor necrosis factor(TNF)-α, interleukin(IL)-6 and IL-1β were detected by Western blot. Results Compared with the control group, HE and VG staining results showed that inflammatory cells gathered and collagen distribution increased in the lung tissue of silicosis mice. Oil red O staining showed that a large number of orange-red lipid droplets appeared in the lung tissue of mice. IHC results showed that the expression of MSR1 was up-regulated in silicosis inflammation stage. Western blot results showed that the expression of MSR1, TNF-α, IL-6 and IL-1β was up-regulated in silicosis inflammation stage(P<0.05). The expression of MSR1 in the SiO2cell stimulation group was up-regulated(P<0.05), and the expression of MSR1 in the siRNA-MSR1 group decreased(P<0.05), and lipid droplets also appeared in the SiO2cell stimulation group. The accumulation of lipid droplets in siRNA-MSR1+SiO2stimulation group was lower than that in SiO2group(P<0.01). ELISA results showed that the expression of TNF-α, IL-6 and IL-1β in SiO2cell stimulation group was up-regulated(P<0.05). Compared with SiO2group, the expression of TNF-α, IL-6 and IL-1β in siRNA-MSR1+SiO2stimulation group was down-regulated(P<0.05). Conclusion MSR1 is involved in the regulation of lipid components and the release of inflammatory factors in lung tissue and cells of silicosis mice. Inhibition of MSR1 expression can antagonize the inflammatory response and abnormal lipid accumulation in macrophages. MSR1 may be a potential therapeutic target for future intervention in the progression of silicosis.

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Objective To investigate the effects of miR-26a-3p on rat myocardial cell(H9c2) injury induced by hypoxia/reoxygenation(H/R) and its mechanism. Methods H9c2 cardiomyocytes in logarithmic growth phase were subjected to hypoxia(1%O2) for 6 h, and reoxygenated at different times(2, 4, 8, 12 h) to establish H/R model cell. Normoxia group was also set up, and cell proliferation activity was detected by cell counting kit-8(CCK-8). The level of lactic dehydrogenase(LDH) in cell supernatant was determined by colorimetry. The expression levels of miR-26a-3p and Survivin mRNA were detected by real-time fluorescence quantitative PCR(qRT-PCR). The expression level of Survivin protein in the cells was detected by Western blot. H9c2 cells were transfected with miR-26a-3p inhibitor and negative control inhibitor NC, Survivin gene siRNA interference plasmid(si-Survivin) and negative control si-NC, followed by H/R intervention. CCK-8 was used to detect cell proliferation in each group. The activity of superoxide dismutase(SOD) and the content of malonaldehyde(MDA) in cell and the level of LDH in supernatant were determined by colorimetry. The apoptosis level of each group was detected by flow cytometry. The protein expression levels of Bcl-2 associated X protein(Bax), B-cell lymphoma-2(Bcl-2), cleaved caspase-3 and Survivin were detected by Western blot. Targeting relationship between miR-26a-3p and Survivin gene was determined by dual luciferase. Results Compared with the normoxia group, proliferative activity, mRNA and protein expression levels of Survivin in H9c2 cells gradually decreased with the extension of reoxygenation time(P<0.05), while LDH and expression level of miR-26a-3p gradually increased(P<0.05). Down-regulating the expression of miR-26a-3p increased proliferative activity, SOD activity, and expression level of Bcl-2 protein in H9c2 cells exposed to H/R(P<0.05), while MDA content, LDH release amount, apoptosis rate, expression levels of Bax and cleaved caspase-3 protein decreased(P<0.05). Survivin deficiency reversed the protective effect of miR-26a-3p inhibitor on H9c2 cells induced by H/R. Dual luciferase reporter gene assay confirmed that Survivin was the target gene of miR-93-5p. Conclusion miR-26a-3p is highly expressed in cardiomyocyte injury induced by H/R. Inhibition of miR-26a-3p expression can inhibit H/R-induced cardiomyocyte apoptosis and oxidative stress by targeted up-regulation of Survivin expression.

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Objective To analyze the epidemiological characteristics of influenza in Anhui province from 2018 to 2023, and to explore the change rule of influenza etiology, so as to provide reference for the prevention and control of influenza. Methods Sentinel hospital surveillance data and network laboratory etiological surveillance data of influenza-like cases(ILI) in Anhui province from 2018 to 2023 were collected by "China influenza surveillance information system", and the data were analyzed using Excel 2019 and SPSS 25.0. Results From 2018 to 2023, ILI% in Anhui province were 3.74%, 4.26%, 3.53%, 4.34%, 6.80%, respectively. The positive rates of influenza virus were 14.87%, 19.98%, 0.12%, 12.39% and 24.72%, respectively. There were significant differences in time distribution and age distribution of ILI% and influenza virus positive rates(P<0.001). Influenza mainly occurred from December to March and from June to August in Anhui province. The positive rate of influenza virus detection was the highest among children under 15 years old, which was the main population of influenza cases. The outbreaks had mainly occurred in primary and secondary schools and childcare facilities. Conclusion There are two incidence peaks of influenza each year, one is in winter and spring, another one is in summer. From December to March, from June to August are the critical periods of influenza control and prevention. Children under 15 years old are the key groups and schools are the key places of influenza control and prevention. Continuous influenza surveillance can provide scientific basis for the prevention and control of influenza in Anhui province.

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Objective To develop a prediction model for live birth based on the basic characters of patients after transcervical resection of adhesions(TCRA) undergoing frozen-thawed embryo transfer(F-ET). Methods A total of 491 cycles who went F-ET after TCRA were included. The cycles were randomly divided into training(n=347) and validation(n=144) cohorts at 7 ∶3 ratio. Relevant factors of live birth were selectedvialogistic regression analysis based on the data from patients in the training cohort. Multivariate logistic regression analysis was used to establish a nomogram, and then this predict model was calibrated and verified. Results Age, body mass index(BMI), miscarriages and endometrial thickness were significantly related to the live birth.The statistical nomogram was built with the area under the ROC curve for the training cohort was 0.713(95%CI: 0.519-0.815), Hosmer-Lemeshow goodness of fit test were χ2=7.062,P=0.530, and the slope of calibration curve was close to 1. AUC of validation cohort was 0.609(95%CI: 0.660-0.561), presenting a well-pleasing goodness-of-fit and stability in this model. Conclusion Age, BMI, miscarriages and endometrial thickness can better predict the probability of live birth undergoing F-ET after TCRA.

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Objective To investigate the differences in gut microbiota and metabolites between urban and rural middle school students and explore their significance in gut homeostasis, so as to establish a healthy lifestyle and diet for children. Methods Fecal samples were collected from middle school students in Hefei(n=14) and Jixi county(n=18, Southern Anhui), aged 13.0-13.5 years. Stool samples were sequenced by 16S ribosomal DNA identification(16S rDNA) and detected for non-targeted metabolites by liquid chromatograph mass spectrometer(LC-MS), followed by bioinformatic analysis. Results LachnoclostridiumandAnaerostipeswere dominant in the urban students that had been reported to be associated with colorectal cancer, atherosclerosis, depression and other disorders. In the village children,Ruminococcaceae UCG-002,BarnesiellaandEubacteriumdominated. An increased proportion of these microbes were related to metabolism of bile acids, short-chain fatty acids, lipid and carbohydrate decomposition, and play an important role in maintaining immune balance and physiological function. Additionally, significant differences in gut metabolites of the two groups were noted, mainly in arachidonic acid metabolism, platelet activation, serotonin metabolism, vitamin absorption, primary bile acid metabolism and other pathways. Conclusion Adolescent students of urban and mountainous areas differ in gut microbiota and metabolites. Rural children have a healthy bacterial flora and metabolites in guts due to a reasonable lifestyle and diet in comparison with the city children.

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Objective To explore the predictive value of oxygenation index(P/F) for clinical outcomes in adult community acquired pneumonia(CAP) patients. Methods 493 patients with CAP eligible for inclusion were extracted from AHCCDM, and were divided into survival group(n=405) and death group(n=88) according to whether they survived at discharge; multivariate Logistic regression analysis was used to identify independent risk factors associated with hospitalization death in CAP patients; the trend of changes in oxygenation index(P/F) and hospital mortality risk was evaluated using a restricted cubic spline(RCS) curve. Results Multivariate Logistic regression analysis found that independent risk factors affecting hospitalization death in CAP patients included acute and chronic health scores(Apache II score)(OR=1.11, 95%CI: 1.07-1.15,P=0.02), P/F(OR=1.70, 95%CI:1.03-2.82,P=0.04), and hemoglobin(Hb) and HCO-3concentrationP>0.05, which were not statistically significant; restrictive cubic spline regression analysis found that there was a non-linear response relationship between P/F and hospital mortality(non-linear testP<0.001), and a P/F of 250 mmHg was the cutoff point for the hospital mortality association analysis. P/F<250 mmHg, for every 1 mmHg increased in P/F, the risk of in-hospital mortality was reduced by 1%,HR: 0.99(0.99-1.00),P<0.001. P/F>250 mmHg, there was no association between CAP patients and in-hospital mortality,HR: 1.00(1.00-1.01),P<0.001, indicating that other factors played a dominant role. Conclusion A significantly reduced P/F has a high predictive value for mortality in patients with community-acquired pneumonia.

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Objective To select and obtainvaginal Lactobacillusstrains with inhibitory effect onGardnerella vaginalis, and to explore the possible mechanism of inhibition ofGardnerella vaginalisbyLactobacillus vaginalisstrains, so as to provide a prerequisite for the development of dominant strains for human disease treatment. Methods Gardnerella vaginalisandvaginal Lactobacillusstrains were isolated from vaginal secretions of patients with bacterial vaginosis(BV) and healthy women, respectively. TheLactobacillusstrains with the ability to inhibit the proliferation ofGardnerella vaginaliswere screened by spot on lawn. Double agar diffusion method was used to compare the inhibitory effects ofLactobacillussolutions, cell-free culture supernatants(CFCs) and bacteria onGardnerella vaginalis. Results Sixty-two strains ofvaginal Lactobacillusstrains were isolated and purified from vaginal secretions of 20 healthy women, belonging to six species. Three strains ofGardnerella vaginaliswere isolated from vaginal secretions of 10 patients with bacterial vaginosis. Among the 62 strains ofvaginal Lactobacillus, 18 strains could inhibit the proliferation ofGardnerella vaginalis. Thevaginal Lactobacillus solutions, the CFCs, and bacteria all had inhibitory effect onGardnerella vaginalis. The inhibitory effects ofvaginal Lactobacillussolutions were higher than those of the CFCs. After the CFCs were treated with proteinase K, the inhibitory effect of eight samples disappeared, and that of 10 samples weakened obviously. Conclusion In this paper, 62vaginal Lactobacillusstrains are purified and identified, of which 18 strains can inhibit the proliferation ofGardnerella vaginalis. The production of antimicrobial protein may be involved in the mechanism thatvaginal Lactobacillusinhibits the proliferation ofGardnerella vaginalis.

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Objective To delineate the similarities and differences among general characteristics and maternal-fetal outcomes in patients with selective intrauterine growth restriction(sIUGR)in monochorionic and dichorionic twin, and to compare the disparate patient types with dichorionic twin sIUGR, with an intent to furnish vital insights for clinical diagnosis and prognostic assessment. Methods Clinical data were collected from 89 patients with twin pregnancies diagnosed with sIUGR. Statistical analyses used thet-test, rank-sum test, and chi-square test. Results Pregnant women with dichorionic twins often leverage multiple assisted reproductive technologies. The maternal outcome for dichorionic twins were significantly later than that for monochorionic twins. Furthermore, the incidence of gestational diabetes mellitus was higher in dichorionic twins compared to monochorionic twins. In terms of fetal outcomes, the proportion of normal neonates with Apgar scores ≥8, body mass, and the incidence of type I sIUGR in dichorionic twin pregnancieswere significantly higher than those in monochorionic twin pregnancies. Conversely, occurrences of umbilical cord sail attachment and a smaller placental share were significantly higher in monochorionic twins. In assessing variations in sIUGR typologies among dichorionic twins, one case of intrauterine death was happened in a pregnancy with type Ⅱ/Ⅲ sIUGR typology, but none in type I. Pregnancies with type I sIUGR typology had a significantly higher proportion of large and small fetuses with Apgar scores ≥8 and better birth quality of smaller fetuses. Conclusions Compared to monochorionic twin pregnancies, there are superior maternal and fetal prognosis for dichorionic twin pregnancies with sIUGR. Clinical management should, therefore, prioritize greater attention to monochorionic twin pregnancies. In addition, this study discovers more complex outcomes in type Ⅱ/Ⅲ pregnancies among dichorionic twins, a subject area currently lacking sufficient domestic research, thereby underlining the need for meticulous monitoring and prompt intervention.