abstract:
Objective To explore the effect of glutamine transporter ASCT2 on myeloproliferative neoplasms(MPN)induced by SHP2 activation mutation, and to observe whether ASCT2 inhibitor l-glutamyl-4-nitroaniline(GPNA) can inhibit the progression of myeloproliferative neoplasms. Methods By constructing MPN mice model induced by Ptpn11E76 K/+activated mutation, the cell proliferation ability, amino acid and glutamic acid content, intracellular glycolysis of normal mice and mutant mice were analyzed, and the expression of ASCT2 protein and energy stress were compared. The mice were treated with ASCT2 inhibitor(GPNA), and ASCT2 protein and energy stress were compared between mutant mice and treated groups. The expression of p-mTOR and related proteins in the energy stress pathway and the rescue effect of the disease were detected. Results Ptpn11E76 K/+Mx1-Cre+activated mutant mice had high expression of glutamic acid,ASCT2 protein and strong cell proliferation ability; ASCT2 inhibitor decreased the progression of myeloproliferative neoplasms. Conclusion The expression level of ASCT2 is closely related to the pathogenesis of myeloproliferative neoplasms induced by SHP2 activated mutation. Inhibition of ASCT2 can effectively decrease the morbidity of myeloproliferative neoplasms, suggesting that ASCT2 may be an effective target for treating this type of disease.

abstract:
Objective To investigate the effects of 4-phenylbutyric acid(4-PBA) inhibiting the endoplasmic reticulum stress(ERS) of hepatoma cells on the antitumor effect of Huaierqing cream. Methods The hepatoma cells were treated with 0(as control), 2, 4, 6 and 8 mg/ml of Huaier extract for 24 h. Then the viability of the cells was determined by MTT. The apoptosis rate of the cells was determined by flow cytometry.The expression levels of the anti-apoptotic proteins Bcl2 and Bax, and the ERS-related proteins GRP78, PERK and IRE1 were determined by Western blot. Then the cells were divided into 4 groups: control group, 4-PBA group, Huaier group, and 4-PBA+Huaier group. After treated accordingly for 24 h, they were determined expression of ERS-related proteins PERK and IRE1, cell viability, apoptosis rate, and expression of Bcl-2 and Bax. Results Compared with the Huaier group, the cells in the 4-PBA+Huaier groups showed significantly lower viability. Apoptosis rates of the HepG2 and Hep3 B cells in the 4-PBA+Huaier group were(1.12±16.98)% and(2.32±15.68)% respectively, while those of the Huaier group were(1.15±12.89)% and(2.58±10.67)% respectively, and the difference was statistically significant(P<0.05). Conclusion The combination of Huaierqing cream and 4-PBA can inhibit the occurrence of ERS in hepatoma cells, inhibit the viability and increase the apoptosis, so as to enhance the antitumor effect of Huaierqing cream.

abstract:
Objective To compare the biological behavior between rhabdomyosarcoma(RMS) fusion gene positive and negative cells, analyze differential miRNAs of RMS fusion gene positive and negative using bioinformatics, and verify the expression of differential miRNAs. Methods The viability, invasion, migration and apoptotic capacity of fusion gene positive RH30 cells and fusion gene negative RD and PLA-802 cells in RMS were detected by CCK-8, Transwell, and flow cytometry, respectively. GEO2 R was used to analyze the differential miRNA of the fusion gene positive and negative cells. Sangerbox was used to analyze differential miRNAs and made Volcano map. qRT-PCR was used to verify the selected differential miRNAs. Results The proliferation ability of RD cells was higher than that of RH30 cells and PLA-802 cells(P<0.05). The invasion, migration and anti-apoptotic ability of RH30 cells were higher than those of PLA-802 cells and RD cells(P<0.05). Volcano plot results showed that the number of up-regulated miRNAs was more than down-regulated miRNAs(P<0.05). According to |log2 FC|>3,P<0.05, 12 differential miRNAs were screened(up-regulated miRNAs contained miR-1, let-7 d-5 p and let-7 b-5 p, down-regulated miRNAs contained miR-196 a-5 p, miR-455-3 p, miR-21-5 p, miR-193 a-3 p, miR-29 b-3 p, miR-29 a-3 p, miR-100-5 p, miR-222-3 p and miR-221-3 p), and qRT-PCR results were consistent with bioinformatics analysis. Conclusion The ability of invasion, migration and resistant apoptosis in RMS fusion gene-positive cells is higher than that of fusion gene-negative cells. The differentially expressed miRNAs between fusion gene positive and negative groups can provide new directions for RMS research.

abstract:
Objective To investigate the molecular mechanism by which short hairpin RNA(shRNA) interferes with silencing of collagen triple helix repeat containing 1(CTHRC1) to induce apoptosis in human breast cancer MCF-7 cells. Methods The shRNA-CTHRC1 plasmid and negative control plasmid were transfected into breast cancer MCF-7 cells. The mRNA and protein expression levels of CTHRC1 were determined by qPCR and Western blot respectively. The cell proliferation activity and apoptosis rate were detected by CCK-8 and flow cytometry respectively. The changes of mitochondrial membrane potential were detected by mitochondrial membrane potential kit(JC-10 staining). The expression of apoptosis-related proteins Caspase 3, Cyt C and p53-mediated mitochondrial apoptosis pathway related proteins p53, Apaf-1, Cleaved-caspase 9, Bcl-2, Bax and PUMA was detected by Western blot. The shRNA-CTHRC1 plasmid and si-p53 interfering plasmids were transfected into MCF-7 cells, respectively or simultaneously, and the expression levels of p53 and Cyt C protein were detected by Western blot, the apoptosis level was detected by flow cytometry. Results Silencing CTHRC1 reduced the mRNA and protein expression levels of CTHRC1, inhibited cell proliferation, promoted cell apoptosis, decreased mitochondrial membrane potential, and upregulated the protein expression levels of Cleaved-caspase 3, p53, Apaf-1, Cleaved-caspase 9, Bax, PUMA and the cytoplasm protein of Cyt C in MCF-7 cells, while down-regulated the protein expression levels of Bcl-2 and the mitochondrial Cyt C in MCF-7 cells. However, interference with p53 expression could reverse the pro-apoptotic effect of CTHRC1 silencing on MCF-7 cells. Conclusion CTHRC1 silencing is dependent on the p53-mediated mitochondrial apoptosis pathway to induce apoptosis in human breast cancer MCF-7 cells.

abstract:
Objective To investigate the effects and mechanism of Icariin on pathological damage and inflammatory response of lung tissue in rats with pulmonary fibrosis. Methods The pulmonary fibrosis model was established with bleomycin(BLM). Rats were randomly divided into healthy control(HC) group、BLM+Icariin(20 mg) group(BI20)、BLM+Icariin(40 mg) group(BI40) and BLM+Icariin(80 mg) group(BI80); HE, TUNEL and Masson staining were used to detect pulmonary pathological injury and fibrosis in rats. Protein expression of Bcl-2, Bax, Caspase-3, Caspase-9, transforming growth factor-β(TGF-β1), α-smooth muscle actin(α-SMA), basic fibroblast growth factor(bFGF), vascular endothelial growth factor(VEGF) and VEGFR was detected by Western blot. Elisa was used to detect the levels of inflammatory factors interleukin-6(IL-6), tumor necrosis factor-alpha(TNF-alpha) and IL-10. Results Icariin could improve the integrity of pulmonary tissue structure and reduce the apoptosis rate(P<0.01), up-regulate Bcl-2/Bax ratio(P<0.01), down-regulate Caspase-3 and Caspase-9 protein expression(P<0.01), up-regulate expression of IL-6 and TNF-α(P<0.01), down-regulate expression of IL-10, inhibit collagen deposition(P<0.01) and the overexpression of bFGF, VEGF and VEGFR. Conclusion Icariin can improve damage of lung tissue and alleviate inflammation in BLM-induced pulmonary fibrosis rats by activating bFGF/VEGF/VEGFR pathway.

abstract:
Objective To investigate the effect of antimicrobial peptide LL-37 on apoptosis of colon carcinoma HT-29 cells and its molecular mechanism. Methods Colon cancer HT-29 cells were treated with antimicrobial peptide LL-37 at different concentrations of 50 μg/ml,100 μg/ml and 200 μg/ml or LL-37 combined with AMPK inhibitor dorsomorphin(10 μmol/L, pre-treated for 30 min) for 48 h. The apoptosis rate of HT-29 cells was assessed by flow cytometry. Apoptotic proteins(Bcl-2 and Bax), autophagy-related proteins(Beclin1, Atg5, LC3 I and LC3 II) and AMPK protein were detected by Western blot assay. Results High dose antimicrobial peptide LL-37 could promote apoptosis of HT-29 cells in colon carcinoma(P<0.05), and increase the expression of autophagy related proteins Beclin1, Atg5 and LC3 II/LC3 I(P<0.05), accompanied by abnormal activation of AMPK(P<0.01). Intervention of AMPK inhibitor dorsomorphin combined with LL-37 could reverse the effect of antimicrobial peptide LL-37 on HT-29 cells in colon carcinoma. Conclusion Antimicrobial peptide LL-37 promotes apoptosis of colon cancer cell line HT-29 by activating AMPK-mediated autophagy.

abstract:
Objective To investigate the relationship between miR-26 b and Wnt/β-catenin signal pathway, different differentiation states of mesenchymal stem cells(MSCs) and its role in the chemotaxis of MSCs to hepatocyte growth factor(HGF).Methods MSCs derived from rat bone marrow were cultured in serum-free induced differentiation medium for 0 d, 7 d, 14 d and 21 d, respectively. The models of MSCs differentiation at different stages of liver-like differentiation were established.The expression of miR-26 b in different differentiated MSCs cells was detected by qRT-PCR. The recombinant adenovirus(Ad-26 b) was used to infect MSCs to achieve high expression of miR-26 b, and the empty virus infected MSCs was used as the control(Ad), the expression of active-β-catenin(ABC), p-β-catenin, β-catenin and the transcription level of Wnt/β-catenin classical target genes c-Myc and RUNX2 were detected. The effects of Transwell and Dunn chamber on the spreading area and adhesion spot of MSCs were determined, and their effects on the chemotaxis and migration of MSCs in different differentiation states were compared.Results With the prolongation of differentiation time, the miR-26 b expression of MSCs increased, and the expression of miR-26 b reached the peak at 14 d after differentiation, and the difference was significant compared with the control group(P<0.05). After high expression of miR-26 b, the level of ABC protein increased,the protein level of p-β-catenin decreased,but the total protein of β-catenin did not change significantly. The expression of downstream target gene c-Myc and RUNX2 of Wnt/β-catenin signal was significantly up-regulated. High expression of mi R-26 b could made the spreading area of MSCs smaller and show polar distribution,and the number of fine adhesive plaques of MSCs increased significantly and distributed to the periphery of the cell front end. Transwell and Dunn chamber experiments showed that the chemotactic migration ability of MSCs to HGF in the induced differentiation group was significantly higher than that in other groups,and the chemotactic migration ability to HGF in the high expression mi R-26 b group was significantly higher than that in the control group( P<0. 05). Conclusion Mi R-26 b affects the differentiation and chemotaxis of MSCs by regulating the Wnt/β-catenin signal pathway.

abstract:
Objective To investigate the change of astrocyte in different regions of immature hippocampus after hyperoxia. Methods In order to set up a model of neonatal hyperoxic immature brain injury, postnatal 6 d rats were taken into 80% oxygen for 24 h as hyperoxia group, and were taken into air room for 24 h as control group. All rats were sacrified for brain tissue at postnatal 7 d, 14 d, and 21 d. Fluorescence intensity of astrocyte in dendate gyrus, hilus and CA1 regions were tested by immunohistochemistry. Results Fluorescence intensity of astrocyte in hilus of hippocampus after 24 h 80% hyperoxia deceased significantly compared with control group at postnatal 21 d. Ratio of fluorescence intensity in cell proliferation and astrocyte in hilus and CA1 regions after hyperoxia deceased significantly compared with control group at postnatal 7 d. Conclusion 24 h 80% hyperoxia inhibited astrocyte growth and proliferation in immature hippocampus.

abstract:
Objective To investigate the mechanism of DNA methylation in papillary thyroid carcinoma, effects of the demethylated drug 5-aza-2′-deoxycytidine(5-Aza-C dR) on the expression level of death-associated protein kinase(DAPK) genes. Methods Human thyroid papillary carcinoma cell line TPC-1 and human normal thyroid cell line Nthy-ori-3 were treated with 10 μmol/L 5-Aza-CdR. Real-Time PCR and Western blot were used to detect the expression levels of DAPK, DNA methyltransferases(DNMTs)mRNA and protein before and after treatment. Results Real-Time PCR results showed that DNMTs mRNA was highly expressed in human thyroid papillary carcinoma cells compared with normal thyroid cells. After 5-Aza-CdR treatment in human thyroid papillary cancer cells, the expression level of DNMTs mRNA decreased, while that of DAPK mRNA increased, and there was significant difference(P<0.05); Western blot results showed that the expression of DNMTs protein was higher in human thyroid papillary carcinoma and DAPK protein expression was less compared with normal thyroid cells, after 5-Aza-CdR was applied for 48 h, the expression of DNMTs protein decreased and the expression of DAPK protein increased(P<0.05). Before and after administration, mRNA and protein expressions of DAPK and DNMTs in normal thyroid cells were not different. Conclusion DNA methylation plays an important role in papillary thyroid carcinoma, 5-Aza-CdR may regulate the expressions of DNMTs mRNA and protein to restore the expression of silenced DAPK gene, which may play a preventive role in thyroid papillary carcinoma.

abstract:
Objective To explore the effects of miR-381 on proliferation and invasion of gastric cancer cells by targeting transmembrane protein 16 A(TMEM16 A) and its mechanism. Methods 34 gastric cancer patients who underwent surgical resection were enrolled. Their cancer tissues and adjacent normal tissues(less than 3 cm away from the cancer) were collected. The gastric cancer cells AGS, MKN45 and SGC7901, and normal human gastric mucosal cells GES-1 were cultured. The expression of miR-381 in tissues and cells was detected by qRT-PCR. The miR-381-mimics and TMEM16 A-siRNA were transfected into AGS cells. The blank control group was set up. The expression of miR-381 and TMEM16 A mRNA in cells was detected. The targeted relationship between miR-381 and TMEM16 A was analyzed by online bioinformatics prediction software and dual-luciferase reporter assay. The cell proliferation was detected by MTT assay. The apoptosis was detected by flow cytometry. The cells invasion ability was detected by Transwell chamber. The expression of p27 gene, cysteine-containing aspartate proteolytic enzyme(caspase-3, caspase-9), E-cadherin, Vimentin and matrix metalloproteinase(MMP-2, MMP-9) in cells was detected by Western blot. Results Compared with normal tissues and normal gastric mucosa cells, the expression of miR-381 in gastric cancer tissues and cells decreased(P<0.05), while expression of TMEM16 A mRNA increased(P<0.05).Compared with blank control group, the expression of miR-381 increased, while the expression of TMEM16 A mRNA decreased in miR-381-mimics group(P<0.05), the expression of TMEM16 A mRNA decreased in TMEM16 A-siRNA group(P<0.05), and the expression of miR-381 increased in combination transfection group(P<0.05). However, there was no significant difference compared with that in miR-381-mimics group(P>0.05). The expression of TMEM16 A mRNA decreased(P<0.05), which was lower than that in miR-381-mimics group and TMEM16 A-siRNA group(P<0.05). Compared with blank control group, cell proliferation in miR-381-mimics group, TMEM16 A-siRNA group and combination group was significantly inhibited, the apoptosis and the expression of p27, caspase-3, caspase-9 and E-cadherin increased, while the number of cell invasion, expression of Vimentin, MMP-2 and MMP-9 decreased(P<0.05). Conclusion MiR-381 can inhibit proliferation and invasion of gastric cancer AGS cells by negatively regulating TMEM16 A, and promote their apoptosis.

abstract:
Objective To explore the effect of apurinic-apyrimidinic endonuclease 1(APE-1) on the tumorigenic ability and PKM2 expression of hepatocellular carcinoma cells in nude mice and its mechanism. Methods Based on silencing APE-1 expression of hepatocellular carcinoma Hep 3 B stable cell line, BALB/c-Nude nude mice were subcutaneously tumorigenic. The APE-1 silencing group was inoculated with APE-1 silenced cells, and the control group was inoculated with cells transfected with blank plasmids, 10 nude mice per group. After 8 weeks, the nude mice were sacrificed, the tumor formation rate was calculated, the tumor tissue was completely excised, the tumor weight was weighed, and the tumor volume was calculated. Immunohistochemistry and Western blot were used to detect the expression of APE-1 and proliferating cell nuclear antigen(PCNA) and Ki-67 in tumor tissues. After detecting the expression of PKM2 protein, based on the data of 374 patients with liver cancer in the TCGA database, the correlation between APE-1 and PKM2 expression was analyzed, and the difference in survival curves of PKM2 expression was compared. Results The tumor formation rate of the nude mice in both groups was 100%. The tumor weight and volume of the APE-1 silenced group were lower than those of the control group(P<0.05). The expressions of APE-1, PCNA, Ki-67 and PKM2 in APE-1 silencing group were lower than those in the control group(P<0.05). There was a linear positive correlation between APE-1 and PKM2 expression(P<0.001), and there was a difference in the overall survival curve between PKM2 in the high expression group and the low expression group(P<0.001). The low expression group had a longer survival time. Conclusion APE-1 may inhibit the tumorigenic ability of hepatocarcinoma cells in nude mice by affecting PKM2, which has strong anti-tumor potential.

abstract:
Objective To study the expression and diagnostic value of miR-1183 in plasma exosomes of colorectal tumors. Methods Electron microscopy, flow cytometry, and nanoparticle tracking analysis(NTA) were used to identify exosomes in plasma samples. miR-1183 expression levels in exosomes of 80 colorectal cancer, 70 colorectal adenoma and 70 healthy control samples were determined by qRT-PCR; the correlation between the expression of exosomes miR-1183 and the clinical data was analyzed by statistics. Results Typical characteristics of exosomes were identified by electron microscopy, flow cytometry, and NTA, it could be considered that plasma exosomes were successfully extracted. The expression level of miR-1183 in plasma exosomes of colorectal tumor group was higher than that of control group; compared with adenoma group, its expression level was significantly higher in colorectal cancer group(allP<0.05). The expression of miR-1183 in plasma exosomes of colorectal cancer group was positively correlated with pathological stage, N stage, M stage, and serum carcinoembryonic antigen(CEA) level(allP<0.05). The expression of miR-1183 in plasma exosomes of colorectal adenoma group was correlated with serum CEA level(P<0.05). As a diagnostic marker, the area under the curve(AUC) of plasma exosomes miR-1183 for colorectal cancer, colorectal adenoma, colorectal tumor, and colorectal benign or tumors were 0.885 7, 0.814 5, 0.852 5, and 0.730 4(allP<0.000 1). Conclusion The expression of exosomes miR-1183 is closely related to the formation and progression of colorectal tumors, and its plasma expression may be used as a marker for auxiliary or hierarchical diagnosis of colorectal tumors.

abstract:
Objective To investigate the effect of miR-455-3 p on the growth and movement of ovarian epithelial cancer cells by targeting HDAC2. Methods Luciferase assay was performed to verify the targeting relationship between miR-455-3 p and HDAC2. HDAC2 pcDNA vector was constructed to overexpress HDAC2 and the cells were transfected into mimic. The cells were randomly divided into four groups: Control group, mimic group, HDAC2 group and mimic+HDAC2 group for subsequent experiments. BRDU staining was used to detect cell proliferation. Cell apoptosis was detected by flow cytometry. The mRNA levels of Ki67, PCNA and c-Myc were detected by RT-PCR. Transwell detected cell invasion. Cell migration was detected by scratch test. Western blot detected vascular endothelial growth factor(VEGF), Vimentin and Fibronectin protein expression levels. Results The results showed that miR-455-3 p had directly targets with HDAC2, compared with HDAC2 group, the number of BRDU positive cells in mimic+HDAC2 group significantly reduced(P<0.05), cell apoptosis rate increased significantly(P<0.05), Ki67, PCNA, c-Myc mRNA level decreased significantly(P<0.05), invasive cells significantly reduced(P<0.05), nick healing rate significantly decreased(P<0.05), VEGF, Vimentin and Fibronectin protein levels significantly decreased(P<0.05). Conclusion miR-455-3 p can induce HO-8910 apoptosis of ovarian cancer cells by targeting HDAC2, inhibit cell proliferation, invasion and migration, and reduce movement ability.

abstract:
Objective To investigate the effect of quinalizarin on cervical cancer Caski cells. Methods Human cervical carcinoma Caski cells were culturedin vitroand pretreated with quinalizarin culture medium at different concentrations(0, 1, 4, 12 μmol/L), 5-fluorouracil group(5 μmol/L) as a positive control. The proliferation of Caski cells was detected by BrdU staining; the apoptosis of Caski cells was detected by flow cytometry; the microtubule-associated protein 1 light chain 3(LC3)content was detected by immunofluorescence; the expression of phosphorylation-related proteins in proliferation, apoptosis, autophagy and PI3 K pathway was detected by Western blot; CaSki cells were injected into nude mice to construct model with the transplanted tumor and verifiedin vivo. Results Compared with the control group(0 μmol/L), the proliferative capacity of cervical cancer CaSki cells was significantly weakened after treatment with 4 μmol/L and 12 μmol/L quinalizarin, the rate of BrdU positive cells significantly decreased, and the expression of Ki67 protein was also significantly declined. With the increase of quinalizarin concentration, the apoptosis rate of cervical cancer CaSki cells and the expression level of Cleaved caspase-3 gradually increased, and the effect was the most obvious in the 12 μmol/L quinalizarin treatment group. With the increase of the concentration of quinizarin, the expression of ATG5 gradually decreased, the ratio of LC3Ⅱ/LC3Ⅰ and the expression of p62 gradually increased, and the 4 μmol/L and 12 μmol/L quinizarin treatment groups had significant differences compared with the control group. Compared with the treatment with the autophagy inhibitor 3-MA alone, the combined treatment of 3-MA and high concentration(12 μmol/L) quinizarin significantly reversed the inhibitory effect of 3-MA on autophagy and the promotion of 3-MA on the phosphorylation of PI3 K pathway.In vivoexperiments, compared with the control group, the tumor type and weight of the drug-treated group significantly reduced, and their Ki67 and ATG5 expressions significantly reduced, and Caspase-3 expression significantly increased, and the phosphorylation levels of PI3 K, AKT, and mTOR protein significantly reduced, which was consistent with thein vitroexperiments. Conclusion Quinalizarin can inhibit the proliferation of cervical cancer CaSki cells, promote autophagy and apoptosis, possibly by inhibiting the phosphorylation of proteins in PI3 K pathway.

abstract:
Objective To preliminary study on the regulation of interleukin(IL)-18 in the expression of tumor necrosis factor alpha(TNF-α), IL-6, and IL-8 in vascular smooth muscle cells(VSMC) mechanism. Methods VSMCs were treated with different concentrations of IL-18( 3,10,30,100 ng/ml),and at different times,TNF-α,IL-6,IL-6,TNF-α,IL-6,Expression of IL-8 m RNA level; ELISA method to analyze the protein level expression of TNF-α,IL-6,IL-8; block p38 and AKT signal pathways with specific inhibitors,and analyze the activation of these two signal pathways by Western blot and its effect on the expression of TNF-α,IL-6 and IL-8 proteins. Results IL-18 at different concentrations could increase the expression of TNF-α,IL-6,IL-8 m RNA in VSMC,showing a dose-dependent effect. Il-18( 30 ng/ml) increased m RNA and protein levels of the three cytokines in a time-dependent manner. Western blot results showed that IL-18 can activate p38 and AKT; analysis of specific blockers found that IL-18 regulates the expression of TNF-α,IL-6,and IL-8 in part dependent on the activation of p38 and AKT signal pathways. Conclusion IL-18 partly relies on the activation of p38 and AKT signaling pathways to increase the expression of TNF-α,IL-6 and IL-8 in VSMC.

abstract:
Objective To explore the role and mechanism of resveratrol(Res) in facial nerve injury model. Methods Male SD rats were randomly divided into sham operation group(Sham group) and facial nerve injury group(Crush group), and facial nerve injury model was established by clamping injury of right facial nerve trunk in rats of the Crush group. The rats in the Crush group were randomly divided into three groups: the control group(Crush+Vehicle group), the resveratrol group(Crush+Res group), and the inhibitor group(Crush+Res+Axitinib group). Western blot and QPCR were used to compare the expression levels of vascular endothelial growth factor b(VEGFb) and its receptors 1 and 2(VEGFR1, VEGFR2) in facial nerve of different groups. Immunohistochemistry was used to evaluate facial nerve regeneration. Results Compared with the Crush+Vehicle group, the expression of VEGFb, VEGFR1 and VEGFR2 in the Crush+Res group was increased and nerve regeneration was promoted. The expression of VEGFb, VEGFR1 and VEGFR2 in Crush+Res+Axitinib group was inhibited compared with that in Crush+Res group. Conclusion Res can enhance the expression of VEGFb, VEGFR1 and VEGFR2 after facial nerve injury and promote nerve regeneration.

abstract:
Objective To explore berberine(BBR) inhibiting apoptosis of nucleus pulposus cells(NPCs) and degradation of extracellular matrix(ECM) in rats induced by H2O2by mediating Notch signaling pathway. Methods NPCs of SD male rats were isolated by sequential enzyme digestion. They were divided into Control group(no intervention),basic fibroblast growth factor( b FGF) group,H2O2group,BBR25 group,BBR50 group and BBR100 group. The proliferation rates in each group were detected by CCK8 method at different time points of incubation. Apoptosis was detected by flow cytometry. The expression of proteins was detected by Western blot. The expression of m RNA was detected by real-time quantitative PCR( RT-PCR). Results After 48 h of cell incubation,cell proliferation rate in bFGF group,BBR25 group,BBR50 group and BBR100 group was higher than that in H2O2group( P<0. 05). Compared with the control group,the expression levels of COL2 A1 m RNA and proteins in H2O2group decreased,while the expression levels of MMP-13 and ACAN m RNA and protein increased. Compared with H2O2group,the expression levels of COL2 A1 m RNA and proteins increased in b FGF group,BBR25 group,BBR50 group and BBR100 group,while the expression levels of MMP-13 and ACAN m RNA and protein decreased( P<0. 05). Conclusion BBR can inhibit NPCs apoptosis and ECM degradation in rats induced by H2O2. The action mechanism may be related to activation of Notch signaling pathway.

abstract:
Objective To investigate the expression of microRNA-34 a(miR-34 a) and silent information regulator 1(SIRT1) in the placentas of patients with pre-eclampsia and the effect of miR-34 a on the expression of SIRT1 as well as the proliferation and apoptosis in HTR-8/SVeno cells. Methods Placental tissues were collected from patients with pre-eclampsia(n=30) and normal pregnant women(n=30). MiR-34 a mimic, miR-34 a inhibitor and SIRT1 siRNA were transfected into HTR-8/SVeno cells. The expressions of miR-34 a and SIRT1 were measured by qRT-PCR and Western blot. Cell proliferation was determined by MTS assay. Apotosis was detected by flow cytometry. Results The relative expression of miR-34 a was significantly elevated and SIRT1 significantly reduced in the preeclamptic placentas. Transfection with miR-34 a mimic in HTR-8/SVeno cells decreased the expression of SIRT1 mRNA and protein. Meanwhile, the rate of proliferation significantly decreased, whereas the rate of apoptosis significantly increased. However, transfection with miR-34 a inhibitor showed the opposite results. Moreover, knockdown of SIRT1 significantly inhibited the rate of proliferation and increased the rate of apoptosis of HTR-8/SVeno cells. Conclusion MiR-34 a may participate in the development of PE by inhibiting the expression of SIRT1 and the proliferation of trophoblasts and inducing the apoptosis of trophoblasts.

abstract:
Objective To investigate the mechanism of quercetin inhibiting proliferation of prostate cancer cells(LNCap) by regulating promoter methylation of glutathione S transferase P1(GSTP1).Methods Human prostate cancer LNCap cells were cultured in vitro,and treated with quercetin( 0,10,40,80 μmol/L) and methyltransferase inhibitor 5'-aza-dc( 10 mol/L) as the control group. After 48 h,CCK-8 method was used to detect the inhibition rate of cell proliferation in each group. Cell apoptosis was detected by flow cytometry. Methylation status of GSTP1 gene was determined by methylation specific PCR( MSP). MRNA and protein expression of GSTP1 in LNCap cells were detected by real-time PCR and Western blot. Results The proliferation rate of LNCap cells decreased with the increase of quercetin dosage and time in a certain concentration range( P<0. 05). The methylation of GSTP1 promoter in LNCap cells decreased( P<0. 05). The expression level of GSTP1 increased( P<0. 05). Moreover,the decrease of GSTP1 m RNA was negatively correlated with the methylation of GSTPl gene( P<0. 05). Conclusion Quercetin inhibited the growth and proliferation of LNCap cells by reducing the methylation of LNCap promoters.

abstract:
Objective To investigate the differentially expressed genes(DEGs) and their functions in endometriosis and to screen potential biomarkers and candidate therapeutic drugs. Methods The stromal cells in ectopic endometrium and normal endometrial tissue were sorted by magnetic beads and analyzed for gene expression profile. Combined with the similar chip data on the same platform available in the GEO database, the DEGs were screened using the limma software package and empirical Bayes method, and GO and KEGG enrichment analysis was performed. The STRING database was queried for PPI interaction network analysis to screen the core pathogenic genes. Then, the CMAP database was retrieved to screen the drug-like small molecules. Results There were significant differences in the expression of 869 genes in ectopic endometrium, of which 17 were up-regulated while 852 were down-regulated. GO enrichment analysis showed that the DEGs mainly regulated receptor ligand activity, channel activity and passive transmembrane transporter activity. KEGG enrichment analysis found that DEGs mostly mediated neuroactive ligand-receptor interaction pathway, cytokine-cytokine receptor interaction pathway and IL-17 signaling pathway. PPI network showed that FPR2, SAA1, RLN3, CXCL8, FPR1, PF4, CCR7 and CXCL5 were the core pathogenic genes. The CMAP database showed that thioronine and flunarizine had potential value in the treatment of endometriosis. Conclusion FPR2, SAA1, RLN3, CXCL8, FPR1, PF4, CCR7 and CXCL5 are potential biomarkers for endometriosis. Liothyronine and flunarizine are drug candidates for the treatment of endometriosis.

abstract:
Objective To evaluate the diagnostic value of acoustic structure quantification(ASQ) technique in chronic hepatitis b cirrhosis and non-alcoholic fatty liver disease. Methods Thirty healthy volunteers, forty patients with chronic hepatitis b cirrhosis and thirty patients with non-alcoholic fatty liver disease were examined by ASQ. The ultrasonic two-dimensional and ASQ images were collected and analyzed. The relevant parameters(redmode, redave, redSD, bluemode, blueave, blueSD and FD ratio)were statistically analyzed. Receiver-operating characteristic(ROC)curves were drew for each parameter. Results The difference of red line parameters was statistically significant between each two groups(P<0.05). BlueSD showed statistically significant differences among all groups except the normal control group and the chronic hepatitis b cirrhosis group(P<0.05), while the difference of bluemode and blueave was statistically significant between each two groups(P<0.05). There were statistically significant differences in FD-ratio between all groups except the normal control group and the non-alcoholic fatty liver group(P<0.05). In chronic hepatitis b cirrhosis group,the ROC evaluation showed that the area under the cure(AUC)of redmode, redave, bluemode and FD ratio(0.940, 0.990, 0.745, 0.920)were greater than 0.7, with higher diagnostic value. The corresponding critical values were 108.32,111.19,15.50,126.84,145.60,26.82 and 0.16 respectively. In the non-alcoholic fatty liver group, the ROC evaluation showed that the AUC of each parameter(0.964,0.951,0.864,0.894,0.852,0.749,0.898)was greater than 0.7, indicating higher diagnostic value. The corresponding critical values were 100.64,104.31,13.95,119.28,126.99,20.08 and 0.09 respectively. Conclusion As a new non-invasive and objective examination method, ASQ technology can not only quantitatively diagnose these two diseases, but also has high value in the diagnosis of non-alcoholic fatty liver disease.

abstract:
Objective To confirm the expression of EPHB2 associated with clinical value and prognosis in endometrial cancer(EC). Methods The role of EPHB2 in ECs was investigated by using publicly available dataset from The Cancer Genome Atlas(TCGA). The correlations between clinical pathologic parameters and EPHB2 were analyzed by the Wilcox test and logistic regression. Then Cox regression and Kaplan-Meier survival analysis were used to analyze the relationships between clinicopathologic characteristics and EPHB2 expression and overall survival in TCGA database. Gene Set Enrichment Analysis(GSEA) was also conducted by TCGA. Results The expression of EPHB2 was increased in EC compared to the normal endometrium. Increased EPHB2 expression in EC was significantly correlated with stage(OR=2.58 for Ⅲ/ⅣvsⅠ/Ⅱ), grade(OR=2.41 for high gradevslow grade), histology type(OR=13.86 for serous/mixed serous endometrioid adenocarcinomavsendometrioid adenocarcinoma) and peritoneal cytology(OR=4.56 for positivevsnegative), etc(P<0.05). Kaplan-Meier methods revealed that patients of EC with high EPHB2 expression had a poor prognosis than those with low EPHB2 expression. The univariate and multivariate analysis showed that EPHB2 could become an independent predictor for poor outcome of EC(P<0.05). GSEA suggested that axon gidance might be enriched in EPHB2 high expression phenotype and α linolenic acid were enriched in EPHB2 low expression phenotype. Conclusion EPHB2 may be associated with development of EC, which could be a potential prognostic molecular marker for poor survival in EC.

abstract:
Objective To study the risk factors of neovascular glaucoma(NVG) and the efficacy of panrentinal photocoagulation(PRP) technology combined with conbercept.Methods 112 patients(112 eyes) with NVG were selected as the disease group, and the healthy people of the same age were selected as the healthy group. The general data of the patients was collected and the logistic regression was used to analyze the risk factors of NVG. The patients were divided into the study group(66 cases) and the control group(46 cases) according to the treatment method. The control group underwent traditional ciliary body cryotherapy, and the study group was treated with intravitreal injection of conbercept and PRP. The effects of different treatment methods on efficacy, intraocular pressure,vision and serum factors were compared. Results The results of logistics regression analysis showed that the levels of serum platelet-derived growth factor-C( PDGF-C),vascular endothelial growth factor( VEGF),tumor necrosis factor-α( TNF-α),and interleukin 6( IL-6) were independent risk factors for NVG. The total effective rate of the study group was higher than that of the control group( 95. 45% vs 82. 61%),and the regression time of the study group was shorter than that of the control group( P<0. 05). After treatment,the intraocular pressure in both groups decreased,and the intraocular pressure level of the study group was better than that of the control group at each time point( P<0. 05). After treatment,the best corrected visual acuity( BCVA) of the patients in the two groups increased,and the BCVA of the study group was higher than that of the control group at each time point( P<0. 05). The levels of serum PDGF-C,VEGF,TNF-α and IL-6 in the study group were lower than those in the control group 1 month after treatment( P<0. 05). Conclusion Serum PDGF-C,VEGF,TNF-α and IL-6 are all risk factors for NVG. The combination of PRP technology and conbercept can improve the patient's intraocular pressure and vision,as well as improve the serum inflammatory factors and angiogenesis factors levels,thereby improving the therapeutic effect.

abstract:
Objective To investigate the clinicopathological features and prognostic factors of small gastrointestinal stromal tumor(GIST). Methods Clinicopathological and mutations features of 34 mini-GIST and 16 micro-GIST patients were retrospectively analyzed. All data were compared with 58 cases of clinically overt GIST with tumor diameter between 2 cm and 5 cm. Results Both the mini-GIST and micro-GIST were predominantly located in the stomach, followed by small intestine and colorectal. The most common pathomorphology of mini-GIST and micro-GIST were spindle cell, mixed cell while epithelioid cell were relatively rare. The mutation rate of KIT in mini-GIST and micro-GIST was lower than that in clinically overt GIST, while the percentage of wild type GIST was significantly higher than that in clinically overt GIST(P<0.05). The mitotic rate, tumor risk grade and Ki67 proliferation index of mini-GIST and micro-GIST were obviously lower than those of clinically overt GIST(P<0.05). However, the synchronous occurrence of digestive tract cancer was much more common in mini-GIST and micro-GIST(P<0.05). Conclusion Most mini-GISTs and almost all micro-GISTs exhibit benign biological behavior and have an excellent prognosis. However, there are still a few mini-GISTs with potential malignancy. Early detection and timely intervention of small GIST with potential malignancy can reduce the risk of recurrence and improve the prognosis.

abstract:
Objective To explore the value of clinical dosage of norepinephrine in evaluating the prognosis of patients with sepsis and septic shock. Methods Retrospective study and analysis of patients with sepsis and septic shock(a total of 300 cases) were admitted. Patients were divided into survival group and death group according to the life and death within 28 d, single-factor analysis was used to screen out the indicators related to the patient′s prognosis, and then multi-factor analysis Logistic regression was used to screen out the independent risk factors which affected the patient′s prognosis. Receiver operating characteristic curve(ROC) curve was used to evaluate the predictive efficacy of the continuous variables of the independent risk factors selected for the patient ′s prognosis, and the results were completed using an area under curve(AUC). The Kaplan-Meier method was used to compare the survival rates of patients in the high-dose group and the low-dose group, respectively. Results (1) Univariate analysis showed there were significant differences in maximum dose of norepine kidney, cumulative dose of norepine kidney, APACHE Ⅱ score, SOFA score, pre-ICU department, basic diseases(respiratory diseases, cirrhosis), ICU average fluid output(P<0.05);(2) the maximum dose of norepine kidney, the cumulative dose of norepine kidney, invasive mechanical ventilation, and APACHE Ⅱ score were independent factors that affect the patients' s poor prognosis;(3) ROC curve display: cumulative dose of norepinephrine was the most effective in predicting theprognosis in patients with sepsis and septic shock;(4) Kaplan-Meier survival curve showed that the survival rate of high-dose group was lower than low-dose group at any time point. Conclusion The dose of norepinephrine administered to patients with sepsis and septic shock can reflect the prognosis of patients.

abstract:
A high-throughput flow cytometry method was established to determine autoantibodies in the plasma of immune thrombocytopenia(ITP) patients and to explore the meaning of clinical significance. The monoclonal antibodies anti-platelet glycoprotein(GP) Ⅸ, Ⅰb, Ⅲa, Ⅱb and P-selectin were coated on five different strength APC-cy7 fluorescent microsbeads for flow cytometry detection. The receiver operating characteristic curve was established to evaluate the specificity and sensitivity of the method as well as the intra-assay CVs and inter-assay CVs differences. The intra-assay CVs were 8.35%, 5.65%, 3.41%, 3.63% and 3.51%, respectively. The inter-assay CVs were 10.10%, 9.13%, 8.04%, 9.59% and 10.20%, respectively. Sensitivity, specificity, and accuracy were 71.0%, 80.4% and 73.5% for anti-GPⅨ; 82.1%, 80.8% and 81.8% for anti-GPⅠb; 71.7%, 79.2% and 73.7% for anti-GPⅢa; 71.0%,76.7% and 72.5% for anti-GPⅡb; 72.0%, 79.2% and 74.0% for anti-P-selectin; 89.0%, 66.5% and 82.9% for the combined detection of the five antibodies. This method was used to detect the level of autoantibody before and after treatment in 75 ITP patients with oral dexamethasone. A new diagnostic method for platelet membrane glycoprotein autoantibodies in patients with immune thrombocytopenic purpura is successfully established based on flow microsphere technology.

abstract:
To collect the clinical data of 306 patients with acute myocardial infarction(AMI) and the occurrence of death during 1 year follow-up. The raw data were randomly divided into modeling group and validation group according to the scale of 7 ∶3. The modeling group used the least absolute shrinkage and selection operator(LASSO) regression and Logistic regression to screen the risk predictors of death at 1 year after operation to construct a nomogram model. The validation group used receiver operating characteristic curve(ROC) curve and Hosmer-Lemeshow deviation test to evaluate the prediction efficiency of the nomogram model. The number of lesions, cystatin C, and left ventricular ejection fraction(LVEF) was the influencing factors of death at 1 year after operation in patients with AMI. The area under the ROC curve of the nomogram model for predicting the risk of death was 0.824(0.621～1.000)(P<0.05), and the Hosmer-Lemeshow deviation test result was χ2=6.090(P=0.637), suggesting that the nomogram model had good discrimination and calibration, and could be used to predict the risk of 1 year postoperative death in patients with AMI.

abstract:
The 7-day-old SPF grade BALB/C suckling mice were gavaged withrotavirus(RV). At the same time, PBS negative control group and normal control group were set up, all mice were kept in SPF grade animal room to observe the diarrhea of suckling mice every day. On the 1 st, 2 nd, 3 rd, 4 th, 5 th, 6 th, 7 th, 8 th, 9 th and 10 th day after inoculation, respectively, 5 suckling mice were killed. The small intestine tissues of suckling mice were collected in sterile environment. The viral content was detected by qPCR method, RV load and the histopathological changes were observed by HE staining, and the activity of lactase was detected by commercial kit. RV was detected in the intestinal tissue of suckling mice on the first day of infection, and the relative content of RV was higher; severe diarrhea occurred in the first 3 d after infection; HE staining showed that the small intestine tissue of suckling mice was damaged, the villi of small intestine were shed off, and the length became shorter. The corresponding results of PBS negative control group and normal control group were negative; the lactase activity was lower than that of the PBS negative control group and normal control group. Therefore, the establishment of RV infected BALB/C suckling mice with secondary lactose intolerance animal model will provide a strong guarantee for studying the pathogenesis of lactose intolerance and drug intervention.

abstract:
ThePSCAgene encoding prostate stem cell antigen(PSCA) andPaFvgene encoding PSCA-specific scFv antibody were amplified and cloned into pET-32 a and pDAP2/S vectors, respectively. After induced for expression of TrxA-PSCA and PaFv-AP fusion proteins, SDS-PAGE and Western blot were conducted to analyze their expression levels and forms, whereas alkaline phosphatase(AP) color-reaction and enzyme-linked immunosorbent assay(ELISA) were applied to detect the activity of PaFv-AP fusion. The results suggested that TrxA-PSCA and PaFv-AP fusion proteins were successfully expressed in soluble form in the constructed prokaryotic expression systems. Moreover, the PaFv-AP fusion retained the antigen-binding capacity of antibody and AP activity.