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Objective To systematically predict the structure and function of theompBgene and its encoded protein from the newly discovered R. Japonica Strain Anhui 120 isolated from the blood of clinical patients in China, so as to provide a theoretical basis for the role in pathogenic mechanisms of rickettsia, the development of diagnostic reagents and vaccines of clinical spotted fever. Methods Based on theompBgene and its encoded amino acid sequence of R.Japonica strain Anhui 120, bioinformatics method was used to predict the signal peptide, transmembrane region, hydrophobicity and possible B cell epitopes. Results There were 15 deletions of consecutive bases of the 3 297～3 311 in theompBgene of R.Japonica strain Anhui 120. In addition, the isolate also had 4 base mutations. The number of amino acids encoded by the gene was 1 651, the molecular weight of the protein was 167.69 ku, and there were 20 kinds of amino acids. The amino acid with more content was Gly. The theoretical isoelectric point pI was 5.15, the extinction coefficient was 58 805 or 58 680, the unstable index was 7.15, the fat index was 88.86, the total average hydrophilicity(GRAVY) was 0.06 and it was hydrophobic protein, the Signal peptide(Sec/SPI) Likelihood was 0.567 7 with signal peptide sequence. There was a Pfam region and autotransporter region, which was associated with bacterial adhesion to host and protein transport respectively. The main secondary structure of the protein was random coils, followed by β-sheet and α-helix, accounting for 53.73%, 30.83% and 15.45%, respectively. In the tertiary structure of homologous model protein, the amino acid of 1 308～1 651 formed a barrel-like structure on the surface of the protein, which might have the action of serine protease. The antigenic epitope in the protein molecule was rich, and there were 159 possible epitopes. The highest predicted score(0.94) was the sequence of 1 269～1 284. Conclusion The composition of theompBgene and its encoded protein, the secondary structure, tertiary structure and B cell epitope of R.Japonica strain Anhui 120 were successfully analyzed and predicted, which provides the theoretical support for the development of pathogenic mechanisms, clinical diagnostic reagents, and subunit vaccines of R.Japonica.

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Objective To investigate the effects of Dendrobium candidum polysaccharide on the differentiation of bone marrow-derived macrophages cells(BMMs) into osteoclasts induced by receptor activator of nuclear factor-κ B ligand(RANKL)in vitro. Methods Water extraction and alcohol precipitation method was used to extract the Dendrobium polysaccharide from Dendrobium candidum. C57 BL/6 mouse BMMs were induced by macrophage colony-stimulating factor(M-CSF). The effects of different concentrations of Dendrobium polysaccharide on the proliferation and cell viability of mouse BMMs were detected by CCK-8 method. The expressions of p65, phospho-p65(p-p65), p38, phospho-p38(p-p38), nuclear factor of activated T-cells cytoplasmic 1(NFATc1) and c-Fos was detected by Western blot. The expression of tartrate-resistant acid phosphatase(TRACP), NFATc1, Cathepsin K(CTSK) and matrix metalloproteinase-9(MMP9) gene were detected by qRT-PCR. The BMMs were induced by RANKL and different concentrations of Dendrobium polysaccharide to carry out TRACP staining. Results When the polysaccharide concentration of Dendrobium candidum reached 400 μg/ml, it significantly inhibited the proliferation of BMMs(P<0.05). Western blot showed that Dendrobium candidum polysaccharide reduced the expression of NFATc1, c-Fos protein, decreased the phosphorylation levels of p65 and p38, which showed a dose-dependent effect(P<0.05). qRT-PCR showed that compared with the RANKL-induced group, the expression of NFATc1, MMP9, CTSK and TRACP genes of Dendrobium polysaccharide group were down-regulated(P<0.05). The TRACP staining showed that the formation of osteoclasts with relatively large volume and>10 nucleus was observed in the RANKL-induced group. The formation of osteoclasts in the concentration of 200 μg/ml was inhibited and only a small number of immature osteoclasts with relatively small volume were formed. Conclusion In vitro, Dendrobium candidum polysaccharide inhibits the expression of osteoclast-associated genes and the synthesis of related pathway proteins, inhibiting RANKL-induced osteoclast differentiation of BMMs.

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Objective To study the effects of dexmedetomidine(DEX) on the excitability of locus coeruleus(LC) neurons in the brain stem in order to elucidate the central mechanisms of the analgesic effects of DEX. Methods The effects of DEX on the mechanical and thermal pain thresholds in adult male C57 BL/6 mice were examined and the sedative behavior scores were evaluated. The effects of DEX on the discharge of LC neurons in free moving mice were measured by multichannel electrophysiologyin vivo. The analgesic and sedative effects of DEX were detected by behavioral tests with intrathecal injection of α2receptor antagonists or morphine receptor antagonists. Results Dex in high dose(12～20 μg/kg) intraperitoneal injected could increase the thresholds of mechanical and thermal pain in mice and they showed significant sedation. Both the sedative and analgesic effects of DEX were dose-dependent. Multichannel electrophysiological recordingsin vivoshowed that the discharge frequencies of LC neurons were increased by DEX in low dose, while they were decreased by DEX in high dose. Intrathecal injection of yohimbine, an α2receptor antagonist, significantly inhibited the analgesic effect of DEX in low dose, but did not affect the sedative effect of DEX in high dose. Intrathecal injection of naloxone, a morphine receptor antagonist, did not affect the analgesic effect of DEX. Conclusion DEX exerts analgesic effects by activating the descending inhibition system of LC, and the spinal α2receptor involves in analgesic effects of DEX. Nevertheless, the sedative effects of DEX are not mediated by the descending inhibition system from LC to spinal cord.

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Objective To investigate the effect of exosomes secreted by glioma stem cells(GSCs) on human-derived brain microvascular endothelial cells(HBMECs) with hypoxia/reoxygenation injury. Methods Human glioma U87 cell line tumor stem cells were cultured in vitro, and the successfully cultured tumor cell spheres were routinely identified by immunohistochemical staining and differentiation. The QIAGEN kit was used to extract exosomes from stem cell culture medium. HBMECs were randomly divided into 3 groups: ① normal control group(CON): 1640 cell culture medium containing 10% exosome-depleted FBS, conventional human brain microvascular endothelium cell culture; ② hypoxia/reoxygenation group(H/R): vascular endothelial cells were given hypoxia for 5 h, and then reoxygenated for 1 h; ③ exosomes pretreated group secreted by stem cells(GSCs-Exo+H/R): HBMECs were incubated with 60 μg/ml of GSCs-Exo prior to H/R treatment. At the end of the treatment, the prolif-eration viability,the migration ability and the apoptosis of the three groups were detected. Results Immunofluorescence staining demonstrated that Nestin and CD133 were specifically expressed in cultured glioma stem cells. Positive expression of GFAP and Neun was observed in differentiation of tumor cells. Compared with the CON group,the proliferation and migration of vascular endothelial cells in the H/R group significantly decreased,and the apoptosis rate significantly increased( P<0. 05). Compared with the H/R group,GSCs-Exo co-incubation with vascular endothelial cells could significantly increase the proliferation of endothelial cells,promote cell migration and inhibit vascular endothelial cell apoptosis( P<0. 05). Conclusion In the hypoxic/reoxygenation microenvironment of glioma,exosomes secreted by glioma stem cells can protect vascular endothelial cells.

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Objective To investigate the role of E2 F transcription factor 1(E2 F1) in hemin-induced primary cortical neuron injuries and the possible mechanisms in rats. Methods Rat primary cortical neurons were cultured, identified and treated with different concentrations of heminin vitro. LDH and MTT assays were performed to detect cell damage when neurons were treated for 12 h. The location and the expression of E2 F1 in neurons were detected by immunofluorescence double staining and Western blot respectively. Rats were divided into vehicle group(DMSO), injury group(Hemin), negative control group(Hemin+Control siRNA) and treatment group(Hemin+E2 F1 siRNA). TUNEL staining and caspase activity were performed in four groups. Results The positive rate of NeuN staining was 95% on the 7 d. With the treatment of hemin(50, 100, 200 μmol/L), the LDH release of neurons increased, cell viability decreased and E2 F1 expression was up-regulated(F=22.9,P<0.05). Compared with the negative control group, LDH release decreased [(73.6±5.2)%vs(42±4.1)%,F=343.4,P<0.05] and the cell viability increased [(28.7±3.4)%vs(61±2.9)%,F=127.7,P<0.05] in the E2 F1 siRNA-treated group. The TUNEL results showed that hemin treatments increased neuronal apoptosis, and E2 F1 siRNA treatments reduced the apoptosis(F=67.2,P<0.05). The caspase activity assay showed that the activities of caspase-3, caspase-8 and caspase-9 in the hemin group increased(t=15.7,t=9.7,t=7.8,P<0.05), but E2 F1 siRNA treatment decreased the activities of caspase-3, caspase-8 and caspase-9(t=8.6,t=7.3,t=5.7,P<0.05). Conclusion E2 F1 siRNA may play a neuroprotective role by inhibiting hemin-induced neuronal apoptosis.

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Objective To investigate the effect and mechanism of microRNA miR-206 on the growth and motility of ovarian cancer cell CAOV3.Methods The targeted gene of miR-206 was predicted by bioinformatics and its targeted-relationship were confirmed by the luciferase reporter assay. Cells were divided into Control, miR-206 mimic,pc-MAPK1 and miR-206 mimic + pc-MAPK1 group,the expression of miR-206 was measured by qRT-PCR,cell growth was measured by Brdu staining,cell invasion was measured by Transwell,the expression of Mitogen-activated protein kinase 1( MAPK1),epithelial cadherin( E-cadherin),vimentin( vimentin),c AMP response element binding protein( CREB),phosphorylated CREB( P-CREB),Ets-like Transcription factor-1( Elk-1),P-Elk-1,proto-oncogene( c-Myc),P-c-Myc and vascular endothelial growth factor( VEGF) were measured by Western blot.Subcutaneous xenograft tumors of nude mice with miR-206 transfected or non-transfected CAOV3 cells were established. The volume of the transplanted tumors was measured. The expression levels of miR-206 and MAPK1 were detected by qRT-PCR. The expression levels of Elk-1,P-Elk-1,c-Myc and Pc-Myc expression was detected by Western blot. Ki67 and vimentin expression were detected by immunohistochemistry. Results miR-206 targeted the inhibition of MAPK1 expression. Compared with the control group,the percentage of Brd U positive cells and the number of invasive cells in the miR-206 mimic group reduced( t = 4. 27,P = 0. 002; t = 3. 43,P = 0. 008),the expression of E-cadherin increased( t = 4. 13,P = 0. 003),P-CREB/CREB,P-Elk-1/Elk-1,Pc-Myc/c-Myc ratios and the expression of vimentin were down-regulated( t = 3. 27,P = 0. 010; t = 3. 26,P = 0. 010; t = 3. 28,P = 0. 010,t =3. 51,P = 0. 007). miR-206 mimic reduced the volume of subcutaneously transplanted tumors of CAOV3 cells in nude mice( t = 6. 51,P<0. 001),increased the expression of miR-206 in the tissues of transplanted tumors( t =4. 65,P = 0. 001),reduced MAPK1 mRNA levels,VEGF expression,P-Elk-1/Elk-1 and Pc-Myc/c-Myc ratio( t =3. 86,P = 0. 004; t = 3. 37,P = 0. 009; t = 3. 34,P = 0. 009; t = 3. 40,P = 0. 008),and reduced the percentage of Ki67 and Vimentin positive cells( t = 4. 45,P = 0. 002; t = 4. 22,P = 0. 002). Conclusion miR-206 over expression can inhibit ovarian carcinoma cell CAOV3 growth and mobility via obstructed activation of MAPK signal pathway.

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Objective To investigate the effect of silenced fibrin original protein 1(FGL1) on the drug sensitivity of gefitinib in lung adenocarcinoma cells. Methods Real-time quantitative PCR and Western blot were used to detect the differential expression of FGL1 in lung adenocarcinoma gefitinib-sensitive cell line PC9 and gefitinib-resistant cell line PC9/GR. FGL1 expression was silenced in gefitinib resistant strain PC9/GR. CCK-8 assay was used to detect cell proliferation in each group. Flow cytometry was used to determine the apoptosis rate of each group. Western blot was used to determine relative expression levels of apoptotic proteins Bcl-2, Bax, Caspase-3 and Cleaved-Caspase-3. Results FGL1 was highly expressed in PC9/GR cells. Silencing FGL1 inhibited the proliferation of PC9/GR and increased the apoptosis rate of PC9/GR cells(P<0.05). Western blot showed no significant difference in the expression of Caspase-3 in each group(P>0.05). Compared with the control group, the expression of Bax and Bcl-2 in gefitinib group showed no significant difference, while the expression of Cleaved-Caspase-3 was up-regulated, with statistically significant difference(P<0.05). After FGL1 was silenced, Bcl-2 protein expression was down-regulated, while Bax and Cleaved-Caspase-3 protein expression was up-regulated, with statistically significant differences(P<0.05). Conclusion Silencing FGL1 can inscrease the sensitivity of lung adenocarcinoma cells to gefitinib, inhibit cell proliferation, promote cell apoptosis, and provide potential therapeutic targets for gefitinib resistant lung adenocarcinoma patients.

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Objective To investigate the protective effect of resolvin D1(RvD1) on experimental colitis and its possible mechanism. Methods Mice were randomly divided into normal control group, RvD1 control group, model control group and RvD1 administration group. The colitis model was constructed by drinking 5% DSS solution freely and RvD1(50 μg/kg) was intraperitoneally injected into mice on 2 nd day, 4 th day and 6 th day. Disease activity index(DAI) score was measured during the experiment. At the end of the experiment, the peritoneal macrophages and the colon tissues were collected. Histological index(HI), myeloperoxidase(MPO), cytokine levels were measured and the structure of autophagosome in peritoneal macrophage was observed by electron microscope. The expression levels of related genes such as Beclin-1, p62, LC3-Ⅱ and pro-IL-1β were detected and analyzed by real-time PCR and Western blot. Results Compared with the model control group, the DAI score, HI score, MPO activity level and the level of proinflammatory cytokine decreased in the RvD1 administration group(P<0.05). The expression levels of Beclin-1, LAMP-2, LC3-Ⅱ/LC3-Ⅰ in peritoneal macrophages increased, and the expression levels of iNOS, IL-1β and p62 decreased(P<0.05). Conclusion RvD1 has a protective effect on experimental colitis, and the mechanism may be involved in the regulation of impaired autophagy in macrophages.

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Objective To investigate the expression of interleukin-11(IL-11) in glioma and its effect on glioma proliferation, migration and invasion. Methods qRT-PCR was performed to determine the expression levels of IL-11 in clinical samples, glioma cell lines and SF126 cells transfected with IL-11 knockdown plasmid. SF126 glioma cells were tansfected with IL-11 knockdown plasmid(shIL-11) or treated with recombinant human IL-11(rhIL-11), and then the colony formation assay was performed to detect the proliferation of glioma cells, and Transwell assay was used to detecte cell migration and invasion. Results The expression of IL-11 mRNA in glioma tissue was higher than that in normal brain tissue. Colony formation assay and transwell assay indicated that knockdown of IL-11 inhibited proliferation, migration and invasion of glioma cells. In contrast, SF126 cells treated with rhIL-11 significantly promoted proliferative, invasive and migratory capacity. Conclusion IL-11 is upregulated in glioma cells and promotes proliferation, migration and invasion ability.

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Objective To study the effects of different iodine nutritional status on methylation of B1 gene of carcinogen B1(BRAF) of rat thyroid sarcoma filter toxic bacteria(V-raf), and to explore whether iodine affects BRAF expression through methylation of BRAF gene. Methods 40 SPF female SD rats of different iodine nutritional status were randomly divided into 5 groups, namely low iodine group(LI), iodine suitable group(NI), and 5 times higher iodine group(5 HI), 10 times high iodine group(10 HI), 20 times high iodine group(20 HI), 8 rats in each group. The model was made by drinking deionized water with different iodine content and feeding low iodine feed. The urine of rats was collected for 24 h to measure urine iodine during the period, and rats were sacrificed after 3 months. Blood samples and thyroid tissues were collected, and the serum free triiodothyronine(FT3) and free thyroxine(FT4) and thyroid stimulating hormone(TSH) levels were determined by chemiluminescence. Bisulfite sequencing PCR(BSP) was used to determine the methylation of CPG island genes in the promoter region. Immunohistochemical technique was used to determine BRAF protein expression. Results ① Compared with NI group, the serum FT3 concentration in LI group and each HI group decreased, and the difference was statistically significant(P<0.05). ② Comparison of the methylation rate of BRAF in the five groups of rats was not statistically significant(P>0.05). ③ The expression of BRAF in thyroid tissues of LI group, NI group and HI group gradually increased, and the difference was statistically significant(P<0.05). Compared with the NI group, the expression of thyroid BRAF in LI group was lower, and the expression in each HI group was higher. Compared with 5 HI group, the expressions of thyroid BRAF in 10 HI and 20 HI group were higher, and the difference was statistically significant(P<0.05). Conclusion Iodine affects the expression of BRAF protein in rats' thyroid gland, but has no obvious effect on the methylation of BRAF gene, suggesting that iodine may affect the function of BRAF gene through other ways.

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Objective To investigate the effects of miR-182 and hypoxia inducible factor-2α(HIF-2α) pathway on the invasion of human extravillous trophoblasts(HTR8/SVneo) during early pregnancy induced by hypoxia. Methods 67 pre-eclampsia(PE) pregnant women were selected as the study subjects, and 84 healthy pregnant women were selected as the control group. All of them underwent cesarean section voluntarily, and placental tissues were collected after operation. HTR8/SVneo cells were culturedin vitroand transfected with miR-182 inhibitor. They were divided into control group, hypoxia group, miR-182 inhibitor group and miR-182 inhibitor negative control group. The expression levels of HIF-2α, vascular endothelial growth factor(VEGF)in placental tissues and HIF-2α, VEGF, E-cadherin, matrix metalloproteinase 9(MMP-9) and MMP-2 in HTR8/SVneo cells were determined by Western blot.The expression of miR-182 in placental tissues and HTR8/SVneo cells was detected by real-time fluorescence quantitative assay(RT-qPCR). Cell apoptosis was detected by flow cytometry, and the invasive ability of cells was determined by Transwell invasion assay in each group. Results Compared with the control group, the expression of HIF-2α and VEGF in placenta of PE patients increased significantly, while the expression of miR-182 decreased significantly(P<0.05). Compared with the control group, the apoptotic rate of hypoxia group increased significantly, the expression levels of invasive ability, miR-182, MMP-9 and MMP-2 decreased significantly, and the expression levels of E-cadherin, HIF-2α and VEGF increased significantly(P<0.05). Compared with the hypoxic group and the miR-182 negative control group, the apoptotic rate of the miR-182 inhibitor group was significantly lower, the invasive ability was significantly higher, the expression levels of HIF-2α, VEGF, MMP-9 and MMP-2 were significantly higher, and the expression levels of miR-182 and E-cadherin were significantly lower(P<0.05). TargetScan database predicted that HIF-2α was the target gene of miR-182, and double luciferase assay confirmed that HIF-2α was the target of miR-182. Conclusion The expression of miR-182 in placenta of PE patients is low, HIF-2α is highly expressed, and miR-182 may enhance the invasive ability of trophoblasts under hypoxia by activating HIF-2α/VEGF signaling pathway.

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Objective To analyze the expression levels of vitamin D(Vit D), liver vitamin D receptor(VDR) in patients with chronic hepatitis B(CHB) and its correlations with liver function and hepatitis B virus(HBV) serological indicators, and further explore whether Vit D can exert antiviral effects in vitro. Methods The serum levels of Vit D were measured in 81 untreated CHB patients and 40 healthy subjects. The expression of VDR in liver tissue was analyzed by immunohistochemistry. HBV-related parameters(HBsAg, HBeAg, HBV DNA) in cell culture supernatants and the protein expressiones(VDR, IRF-9, MxA) in cells were detected after HepG2.2.15 cellswere treated with Vit D alone or in combination with IFN-α． Results The serum Vit D level in CHB group was lower than that in healthy controls( HC) group( P<0. 05). The serum Vit D level was positively correlated with albumia( ALB)( r = 0. 339,P = 0. 002),negatively correlated with HBV DNA viral load( r =-0. 274,P =0. 013),correlated with e atigen status( P<0. 05),no correlation with alanine aminotransferase( ALT) 、aspartate aminotransferase( AST) 、total bilirubin( TBIL) 、alkaline phosphatase( ALP) 、gamma-glutamyl transferase( GGT),HBsA g and HBc Ab. Also,the expression levels of Vit D and VDR were related to the severity of CHB. The expression levels of Vit D in serum and VDR in liver tissue of severe CHB patients were lower than those of mild and moderate CHB patients. The expression of VDR,JAK-STAT signal transduction molecules IRF-9 and downstream antiviral protein Mx A in the Vit D combination with IFN-α group were higher and the secretion of HBV-related antigens was lower than those in the blank group and the single treatment group. Conclusion There may be a decline in the function of the Vit D/VDR signaling pathway in patients with CHB. Vit D may play a synergistic IFN-α antiviral effect by increasing the VDR expression and thereby enhancing the IFN-JAK-STAT signaling pathway.

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Objective To investigate the effect of exosomes(Exo) released from adipose derived stem cells(ADSCs) with overexpressed miR-34 a on proliferation and apoptosis of human hypertrophic scar fibroblasts(HSF). Methods Primary ADSCs were isolated from human adipose tissue and their surface markers were identified by flow cytometry. The osteogenic differentiation of ADSCs was observed by using alizarin red and alkaline phosphatase staining. ADSCs were infected by lentiviruses containing miR-34 a to obtain over-expressed miR-34 a ADSCs(ADSCs/miR-34 a) and negative control ADSCs(ADSCs/miR-NC). The exosome was extracted from cell supernatant of each group and ADSCs exosome(ExoADSCs), over-expressed miR-34 a ADSCs exosome(ExoADSCs/miR-34 a) and negative control ADSCs exosome(ExoADSCs/miR-NC) were obtained. The morphology of exosomes was observed by transmission electron microscope(TEM), and the surface marker proteins CD81 and CD63 were detected by Western blot. The exosomes were co-cultured with HSF and divided into control group(HSF+PBS), Exo ADSCs treatment group(HSF+ExoADSCs), ExoADSCs/miR-NCtreatment group(HSF+ ExoADSCs/miR-NC) and ExoADSCs/miR-34 atreatment group(HSF+ExoADSCs/miR-34 a). Quantificational real-time PCR(qRT-PCR) was used to detect the expression of miR-34 a in ADSCs and exosomes. The proliferation of HSF cells was detected by MTT assay. The apoptotic rate of HSF cells was detected by flow cytometry. The expression levels of apoptosis related proteins(cleaved-Caspase-3, Bax and Bcl-2), fibrosis-related proteins(COL Ⅰ, COL Ⅲ) and TGF-β/Smad signal pathway related proteins(TGF-β1, p-Smad 2 and p-Smad3) were detected by Western blot assay. Results ADSCs were successfully isolated and possessed osteogenic differentiation ability. TEM showed round or oval vesicle bodies with a diameter of 50～100 nm with bilayer membrane structure. Western blot showed positive expression of CD81 and CD63, which were the marker proteins of exosomes. Overexpression of miR-34 a could increase the expression of miR-34 a in ADSCs and exosomes. Compared with ExoADSCsgroup and ExoADSCs/miR-NCgroup, ExoADSCs/miR-34 aintervention could inhibit HSF cell proliferation(P<0.05), increase cell apoptosis rate(P<0.05), up-regulate the expression of cleaved-Caspase-3 and Bax protein(P<0.05), and down-regulate the expression of Bcl-2, COL Ⅰ, COL Ⅲ, p-Smad2, p-Smad3 and TGF-β1 protein(P<0.05). Conclusion The exosome of miR-34 a overexpressed ADSCs may inhibit the HSF cell proliferation and promote cell apoptosis by down-regulating TGF-β/Smad signaling pathway.

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Objective To investigate the protective effect of emodin on cardiac function in rats with myocardial ischemia-reperfusion injury(MIRI) by activating Nrf2/ARE/HO-1 signaling pathway. Methods Rats were randomly divided into Sham group, MIRI group, Positive control group, emodin 20, 40, 60 mg/kg group, then MIRI model of rats were established, ECG detection of left ventricular end-diastolic pressure(LVEDP), left ventricular systolic pressure(LVSP), maximal rise/fall rate of left ventricular pressure change(±dp/dtmax), myocardial infarct size was measured, biochemical analyzer was used to detect the concentration of serum creatine kinase(CK), lactate dehydrogenase(LDH) and troponin(cTnI), myocardial tissue pathological changes were observed by HE staining, cleaved Caspase-3 positive cell rate was detected by immunohistochemistry, the content of myeloperoxidase(MPO), malondialdehyde(MDA), Superoxide dismutase(SOD) was measured by kits, Western blot was used to detect the protein levels of nuclear factor E2 related factor 2(Nrf2) and heme oxygenase 1(HO-1). Then rats were randomly divided into Sham group, MIRI group, emodin group and emodin+ML385 group, rats in emodin+ML385 group were treated by Nrf2 inhibitor ML385, and changes of cardiac function, pathological changes, inflammation and oxidative stress were detected. Result Compared with Sham group, the rats in MIRI group had elevated LVEDP and LVSP, decreased ±dp/dtmax, increased myocardial infarct size, increased serum CK, LDH, and cTnI concentrations, obvious pathological changes in tissues, and the positive cell rate of cleaved Caspase-3 increased, the MPO and MDA cotent increased, SOD content decreased, protein levels of Nrf2 and HO-1 increased. Emodin treatment played a protective role for cardiac function damage, Nrf2 signaling pathway was further activated. ML385 treatment significantly inhibited emodin induced protective effect of cardiac function. Conclusion Emodin can alleviate MIRI injury, and its mechanism is related to the activation of Nrf2/ARE/HO-1 signaling pathway.

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Objective To investigate the effect of protein arginine methyltransferase(PRMT1) on gene expression of PI3 K-AKT pathway in gastric cancer. Methods The differentially expressed genes between 2 cases of gastric cancer and 2 cases of adjacent tissues were screened by transcriptome sequencing. The key genes affecting the pathway of gastric cancer were enriched and analyzed by kyoto gene encyclopedia of genome(KEGG) and verified by immunohistochemistry. Results 758 differentially expressed genes with length greater than 200 bp were obtained by high throughput sequencing. KEGG enrichment analysis revealed 25 significant enrichment metabolic pathways, of which PI3 K-AKT metabolic pathway was significantly abnormally activated containing 15 genes significantly up-regulated. cBioportal database correlation analysis showed that PRMT1 was negatively correlated with the expression of key genes(PPP2 R3 A, THBS4) in PI3 K-AKT signaling pathway. Immunohistochemical results further showed that there was a high expression of p-AKT(473 Ser) in gastric cancer tissues with low PRMT1 expression, moreover, there was a negative correlation between PRMT1 and p-AKT(473 Ser), indicating that PRMT1 was negatively correlated with PI3 K-AKT signal activity. Conclusion PRMT1 may affect the activity of PI3 K-AKT signaling pathway in gastric cancer through PPP2 R3 A and THBS4, which may provide clues for finding diagnostic markers and therapeutic targets of gastric cancer.

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Objective To explore the effect of exogenous recombined-growth differentiation factor 11(r-GDF11) on calcification of arteries in SD rats. Methods The SD rats were divided into five groups: calcification group, solvent control group, low-concentration group, medium-concentration group and high-concentration group. After 1 week of adaptive feeding, intraperitoneal injection of normal saline, r-GDF11 solvent and low, medium, high concentration r-GDF11 were given daily for 7 weeks, and then calcification was introduced. After 7 weeks of calcification induction, blood was collected from each group and the aortic vessels were separated. Von kossa·HE staining was used to observe the pathological status of vascular calcification. ELISA was used to determine the GDF11 and bone morphogenetic protein 4(BMP4) level in serum. Western blot and immunohistochemistry were used to determine the levels of calcified markers like BMP4 and runt-related transcription factor 2(RUNX2) and the distribution of GDF11 in blood vessels. Results ① The serum BMP4 and GDF11 levels in the calcification group and the solvent control group were significantly higher than those in the low, medium and high concentration group, and the serum BMP4 and GDF11 levels in the low, medium and high concentration group successively decreased. ② Compared with the calcification group and the solvent control group, the calcium salt deposition in the low, medium and high concentration groups decreased, and the elastic fiber was more complete. The higher the r-GDF11 concentration, the less the calcium salt deposition and the more complete the elastic fiber. ③ The expression of RUNX2 and BMP4 in the low, medium and high concentration groups was lower than that in the calcification group and the solvent control group. The higher the r-GDF11 concentration, the lower the expression of calcification marker protein and the rarer the distribution of calcification marker protein in the immunohistochemistry. The higher the concentration of r-GDF11, the higher the expression of GDF11 in tissues and the more intensive the distribution in immunohistochemistry. Conclusion Exogenous treatment with r-GDF11 can reduce the effect on artery calcification in SD rats.

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Objective To analyze the functional changes of Arg 307>Gln mutant of purinergic receptor P2 X ligand-gated ion channel 7(P2 X7 R) in high uric acid background by establishing a human leukemic monocyte(THP-1) derived monocyte cell line in wild type or different mutants to express P2 X7 R stably. Methods THP-1 cells were transfected by lentivirus to establish a THP-1 cell line that stably expressed wild type, Arg 307>Gln mutantand empty virus. Then three types of wild type,Arg 307>Gln mutant and empty virus were set up separately,and each type was randomized into three groups: monosodium urate( MSU)( Group M),MSU and adenosine triphosphate( ATP)( Group MA) and unstimulated control group( Group C). Three groups of cells were stimulated by Phorbol-12-myristate-13-acetate( PMA),then PMA was removed by centrifugation. Group M and MA were added with MSU incubation,then ATP was added to MA group. Three groups of serum and cells were collected. Interleukin-1β( IL-1β) protein expression level in serum was detected by ELISA while mRNA expression of IL-1β,oligomerization domain( NOD)-like receptor family,pyrin domain containing 3( NLRP3),apoptosis-associated speck-like protein containing a caspase-1 recruitment domain( ASC) and Caspase-1 in transfected THP-1 cells was detected by qRT-PCR. Results There was no significant difference between wild type,Arg 307>Gln mutant and empty virus after adding MSU crystals alone( P>0. 05). However after incubation with ATP,the levels of IL-1βand NLRP3 in the wild type were significantly higher than those in the Arg 307>Gln mutant,and the difference was statistically significant( P<0. 05). The mRNA levels of ASC and Caspase-1 in the ATP-P2 X7 R-IL-1β signaling pathway were not significantly different between wild type and Arg 307>Gln mutant( P>0. 05). Conclusion Arg 307>Gln mutant significantly down-regulated the function of P2 X7 R and reduced the expression levels of IL-1β and NLRP3. Caspase-1 and ASC may be degraded during the activation pathway under high uric acid background.

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Objective To study the effects of Hashimoto′s thyroiditis(HT) on the content of serotonin(5-HT) and the distribution of serotonin 2 c receptor(5-HT2 CR) in hippocampus of early pregnant mice. Methods 70 NOD female mice were randomly divided into normal pregnancy group(CON group) and HT pregnancy group(HT group).The HT mouse model was established by 2 times induction of immunization with porcine thyroglobulin(pTg) plus Freund′s adjuvant. Then they were caged with male rats and ended at 4.5 d of pregnancy. Each group randomly selected 16 pregnant rats. Serum thyroid hormone and antibody levels were determined by electrochemiluminescence immunoassay(ECLIA) and enzyme-linked ELISA. Thyroid was stained by HE staining. The content of 5-HT in hippocampus of mice in each group was measured by ELISA. The content and distribution of 5-HT2 CR in hippocampus of mice in each group were observed by Western blot and immunohistochemistry. Results Compared with the CON group, the levels of thyroid peroxidase antibody(TPOAb) and anti-thyroglobulin antibody(TGAb) in the HT group increased(P<0.05), and there was no statistically significant difference in free triiodothyronine(FT3), free tetraiodothyronine(FT4) and thyrotrophin(TSH). HE staining showed that some thyroid follicles were destroyed and lymphocytes infiltrated in the thyroid gland of mice in HT group than those in CON mice. The expression of 5-HT in hippocampus of HT group was lower than that of CON group(P<0.01). The expression level of 5-HT2 CR of HT group was higher than that of CON group. The increase(P<0.001) was mainly distributed in the pyramidal cell layer of hippocampal CA3 region(P<0.01), and the CA1 and DG regions showed an increasing trend. Conclusion HT leads to the decrease of 5-HT expression and the increase of 5-HT2 CR expression in hippocampus of early pregnancy mice.

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Objective To investigate the effects and mechanism of pretreatment with rosiglitazone(RSG) on lipopolysaccharide(LPS)-induced acute liver injury(ALI). Methods 42 male CD-1 mice were randomly divided into six groups(7 mice every group): Control group, RSG group, LPS 6 h group,LPS 12 h group,RSG+LPS 6 h group, RSG+LPS 12 h group. Mice were supplemented with RSG(10 mg/kg) by gavage five consecutive days before LPS solely injection in the RSG and RSG+LPS groups. All mice were singly intraperitoneal injected with LPS(2 mg/kg) in the LPS and LPS+RSG groups. In the Control group and RSG group, mice were intraperitoneal injected with equal normal saline. All mice were sacrificed at 6 h and 12 h after LPS injection. Mice livers were collected and weighted. Some livers were used for HE staining to observe inflammatory cells infiltration and pathological damage. Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected by Reitman. Serum tumor necrosis factor alaph(TNF-α) and macrophage inflammatory protein-2(MIP-2) were measuredviaELISA.The protein expression of activator protein 1(AP-1) signaling, NADPH oxidases(NOX) and antioxidant enzymes were detected using Western blot. Results RSG pretreatment alleviated LPS-induced inflammatory cells infiltration and pathological damage. RSG pretreatment inhibited LPS-induced upregulation of serum ALT and AST in mice. RSG pretreatment attenuated LPS-induced hepatic c-fos and c-jun phosphorylation. RSG pretreatment mitigated LPS-induced upregulation of hepatic NOX-4. RSG pretreatment attenuated LPS-induced upregulation of hepatic haem oxygenase 1. Conclusion RSG pretreatment prevents LPS-induced ALI partially through inhibiting hepatic AP-1 signaling and NOX, and regulating antioxidant enzyme.

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Objective To study the effect of S component of panton-valentine leucocidin(LukS-PV) on proliferation of hepatocellular carcinoma(HCC) cells and its mechanism. Methods HepG2 and Huh7 cells were treated with LukS-PV at different concentrations(0.25,0.5,0.75,1 μmol/L).Cell proliferation was evaluated by CCK-8 method.Changes in overall acetylation level after LukS-PV treatment were detected by pan-acetylated antibody,and changes in protein expression profile of HepG2 cells after LukS-PV treatment were detected by quantitative proteomics.TCGA database was used to analyze the expression of histone deacetylase 3(HDAC3) in HCC and its prognostic correlation.RT-PCR and Western blot were used to verify the changes of HDAC3 mRNA and protein levels in HepG2 and Huh7 cells treated with different concentrations of LukS-PV.Changes in proliferation capacity of HCC cells were detected after HDAC3 knockdown with specific SiRNA. Results LukS-PV could decrease the proliferation ability of HCC cells in a time-dependent and concentration-dependent manner.LukS-PV could increase the acetylation level of HepG2 cells,and quantitative proteomics results showed that HDAC3 expression was down-regulated in HepG2 cells after LukS-PV treatment.TCGA data analysis showed that the expression level of HDAC3 mRNA in liver cancer tissues was higher than that in normal tissues,and the survival time of patients with high HDAC3 expression was lower than that of patients with low HDAC3 expression.The mRNA and protein levels of HDAC3 were down-regulated by LukS-PV in HepG2 and Huh7 cell lines.Knockdown of HDAC3 reduced the proliferation of HepG2 and Huh7 cells. Conclusion LukS-PV may inhibit the proliferation of HCC cells by down-regulating the expression of HDAC3,which provides a new research idea for the precise and targeted treatment of HCC.

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Objective To elucidate the role of long non-coding RNA(lncRNA), a novel tumor regulator, PVT1 in hepatocellular carcinoma(HCC). Methods According to the analysis of TCGA database, PVT1 was highly expressed in HCC. Transient transfection was used to construct PVT1 knockdown cell lines. CCK-8 assay and scratch assay were used to evaluate the ability of cell growth and metastasis. Bioinformatics prediction and luciferase assay were used to validate the targeted binding of PVT1 to miR-17-5 p. The co-transfection experiment was used to verify the effect of PVT1 on proliferation and metastasis through miR-17-5 p. Results The results showed that PVT1 was highly expressed in HCC tissues and cell lines. Knockdown of PVT1 decreased the ability of cells proliferation and metastasis. Luciferase assay confirmed that PVT1 could bind to miR-17-5 p, and miR-17-5 p mediated the effect of PVT1 on the proliferation and metastasis of HCC cells. Conclusion lncRNA PVT1 can enhance the proliferation and metastasis of HCC cells through miR-17-5 p. The above results indicate the important role of PVT1 in HCC and provide a good prospect for future targeted therapy.

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Objective To investigate the effect of overexpression of a disintegrin and metalloproteinase 10(ADAM10) on myocardial cell apoptosis and Notch/Akt signaling pathway in acute myocardial infarction(AMI) rats. Methods A total of forty SD rats were randomly divided into four groups:sham operation group(sham), acute myocardial infarction group(AMI), empty adenovirus group(Ad-NC) and ADAM10 adenovirus group(Ad-ADAM10), with 10 rats in each group. Except for the sham group, the other groups were ligated the left anterior descending branch of coronary artery to establish the AMI models, and adenovirus was injected to the myocardial infarction area in situ. Cardiac function was detected by cardiac ultrasound after 72 h of intervention. Myocardial tissue was taken and TCC staining was used to evaluate myocardial infarction area. TUNEL staining was used to detect myocardial cell apoptosis. ADAM10 mRNA expression level in myocardial tissue was detected by RT-PCR. The expression levels of ADAM10, apoptosis-related proteins and Notch/Akt signaling proteins were detected by Western blot. Results Compared with sham group, the cardiac function in the AMI group decreased(P<0.05), myocardial infarct size and myocardial apoptosis rate increased(P<0.05), the mRNA and protein expression of ADAM10 and the protein expression levels of Bcl-2, Notch1, Hes1 and p-Akt decreased(P<0.05), while the protein expression levels of Cleaved-caspased-3 and Bax increased(P<0.05). Compared with AMI group, the cardiac function of rats in Ad-ADAM10 group improved(P<0.05), myocardial infarct size and myocardial apoptosis rate decreased(P<0.05), the mRNA and protein levels of ADAM10 in myocardial tissue and the protein expression of Bcl-2, Notch1, Hes1, p-Akt increased(P<0.05), while the protein expression levels of Cleaved-caspase-3 and Bax decreased(P<0.05). However, there was no significant difference between the above indexes in Ad-NC group and AMI group(P>0.05). Conclusion Overexpression of ADAM10 can improve cardiac function and inhibit cardiomyocyte apoptosis in AMI rats, and its mechanism may be related to the activation of Notch/Akt signaling pathway.

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Objective To investigate the effects of three different doses of ethanol extracts from Schefflera arboricola Hayata flower on central and cardiac toxicity induced by bupivacaine in rats. Methods Sixty 10-week-old male SD rats were randomly divided into four groups, 15 in each group. After fasting for 12 h, all rats were given intragastric administration once a day for 3 consecutive days. The control group(group D) was oral administrated with normal saline, and the other three groups were oral administrated with ethanol extract of Schefflera arboricola Hayata flower 2 g(group S1), 4 g(group S2) and 8 g(group S3) per kilogram rat, respectively. Then all rats were intravenous pumped with 0.5% bupivacaine 2 mg/(kg·min). The dosage of bupivacaine was recorded when convulsion, ventricular arrhythmia and cardiac arrest occurred. The blood gas analysis of rats in each group was executed when convulsion occurred. Heart tissue HE section pathological examination was executed. The expression of NMDA receptor 1 and 2 B subunits in hippocampus of rats in each group was measured by qRT-PCR. Results The dosage of bupivacaine in group S1, S2 and S3 was higher than that in group D when convulsion, arrhythmias and cardiac arrest occurred, and the dosage of bupivacaine in group S2 and group S3 was also higher than that in group S1. The differences were both statistically significant(P<0.05). Blood gas analysis showed that PaCO2of rats in group S1, S2 and S3 was higher than that of rats in group D during convulsion, and the difference between group S3 and group D was statistically significant(P<0.05). Histological examination of HE section showed that inflammatory cells infiltrated, interstitial congestion or hemorrhage was obvious in group D. Inflammatory cells infiltration in group S1, S2 and S3 was significantly less than that in group D, and interstitial congestion or hemorrhage was slight. The expression levels of NMDA receptor 1 subunit and 2 B subunit in group S1, S2 and S3 were lower than those in group D by qRT-PCR analysis, and the difference was statistically significant(P<0.05). Conclusion The central and cardiac toxicity in rats induced by bupivacaine could be alleviated by pre-administration of the ethanol extract of Schefflera arboricola Hayata flower.

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Objective To investigate whether the polymorphism of tryptophan hydroxylase 2(TPH2) gene is associated with erectile dysfunction(ED). Methods A total of 59 men diagnosed with ED and 151 healthy controls were recruited. International index of erectile function-5(IIEF-5) and the premature ejaculation diagnostic tool(PEDT) data were collected. All ED patients and healthy controls were genotyped for polymorphisms in the TPH2 gene. Allele and genotype frequencies of single nucleotide polymorphisms(SNPs) were compared between the patients and controls. Result The rs1007023 G allele frequency was significantly higher in ED patients than that in the control group(P=0.023). Conclusion The SNPs rs1007023 of the TPH2 gene seems to be associated with ED in males with the rs1007023 G allele or G/G+T/G allele in the Han population.

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Objective To investigate the correlation between plasma orexin-A and leptin levels and obesity in hospitalized patients with schizophrenia. Methods A total of 322 subjects were included, who met the diagnostic criteria of ICD-10 for schizophrenia. The 30-items positive and negative syndrome scale(PANSS-30) was used to evaluate the clinical symptoms of patients. The plasma orexin-A and leptin concentrations of the patients were determined by the ELISA method, and all data were analyzed by SPSS(version16.0). Results Compared to inpatients without obesity, the levels of plasma orexin-A and leptin were significantly higher(P=0.012,P<0.001) respectively. BMI of inpatients with obesity was positively correlated with orexin-A(r=0.22,P<0.001), leptin(r=0.57,P<0.001), triglyceride(TG)(r=0.41,P<0.001) and low density lipoprotein(LDL-C)(r=0.17,P=0.002), and negatively correlated with high density lipoprotein(HDL-C)(r=-0.23,P<0.001). Binary logistic regression analysis showed that high levels of leptin(OR=1.532, 95%CI: 1.260～1.846,P<0.001), orexin-A(OR=1.271, 95%CI: 1.026～1.575,P=0.028) and TG(OR=1.253, 95%CI: 1.029～1.525,P=0.025) were risk factors of obesity in patients with schizophrenia, while higher level of HDL-C(OR=0.207, 95%CI: 0.058～0.739,P=0.015) was a protective factor of obesity. Conclusion High levels of orexin-A, leptin and triglyceride may be risk factors of obesity in hospitalized patient with chronic schizophrenia.

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Objective To investigate the clinical features and prognostic factors of primary gastrointestinal lymphoma(PGIL). Methods Clinical data of 216 patients with PGIL diagnosed by endoscopic biopsy or surgical pathology were collected, and clinical characteristics, endoscopic manifestations and their effects on prognosis were retrospectively analyzed. Results The average age of 216 patients with PGIL was 57.94 years and the ratio of male to female was 1.37 ∶1. There was no specific clinical symptoms. Lesions tended to occur in the stomach, followed by ileocecal region, ileum, colon, jejunum, rectum and duodenum. 23.15% cases invaded multiple parts of the digestive tract. The three most common pathological types were diffuse large B-cell lymphoma(DLBCL)( 69.4%), mucosa-associated lymphoid tissue lymphoma(MALT)(13.0%) and mantle cell lymphoma(MCL)(6.0%). Primary gastric lymphoma(PGL) and primary intestinal lymphoma(PIL) had statistical differences in gender, Lugano staging and the depth of lesion(P<0.05). Endoscopic characteristics were not significantly different between DLBCL, MALT lymphoma and MCL in PGL, which were mostly ulcerative-type. In PIL,DLBCL mostly presented as protuberant but MCL was more inclined to be polypoid. Univariate analysis showed the age>60 years, low albumin(Alb), high beta-2 microglobulin(β2-MG) and lactic dehydrogenase(LDH) levels, Lugano staging IIE and Ⅳ, tumor size ≥10 cm, breaking through serosa, and T/NK cell lymphoma suggested poor prognosis of PGIL. Multivariate analysis showed that age and Lugano staging were independent factors for the prognosis of PGIL. Conclusion PGIL is more common in middle-aged and elderly men, usually in the stomach and ileocecal region. DLBCL is the most common pathological type. Under endoscopy, PGL tends to be ulcerative and PIL is mostly protuberant. PIL has different endoscopic manifestations according to different pathologic types. Age, Alb, β2-MG, LDH, Lugano staging, tumor size and depth,histological subtype are all factors affecting the prognosis of PGIL.

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Objective To evaluate the value of noninvasive novel indicators fibrosis index based on four factors(FIB-4)、aspartate transaminase-to-platelet(APRI) and Gamma-glutamy transpeptidase-to-platelet ratio(GPR) in differentiating liver inflammation and fibrosis in antiviral treatment of HBeAg-negative chronic HBV infected patients. Methods Retrospective analysis was performed on 78 patients with HBeAg-negative chronic hepatitis B virus(HBV) infection. The results of liver biopsy and laboratory examination were recorded.Univariate and multivariate logistic regression analysis were used to judge the independent predictor of antiviral treatment indications for HBeAg negative chronic HBV infection. The receiver-operating characteristics(ROC) curve was applied to compare the value of the independent predictive index in evaluating the antiretroviral treatment indications of liver inflammation and fibrosis degree of HBeAg-negative.ROC analysis was also used in validation group composed of 77 HBeAg-negative chronic HBV infected patients who also met the inclusion criteria.The patients whose liver pathology indicated significant inflammation or significant liver fibrosis(inflammatory activity≥G2 or fibrosis stage≥S2)were divided into Inflammatory activity≥G2 and Fibrosis stage≥S2 group and only Inflammatory activity≥G2 or only Fibrosis stage≥S2 group respectively. The independent predictive index was put into the two groups for evaluation using ROC analysis. Results After univariate and multivariate logistic regression analysis, FIB-4 and lgHBsAg were assured that they were the independent predictors of anti-viral treatment indications for HBeAg-negative chronic HBV infected patients.OR(95%CIvalue) were 4.083(1.748～9.538) and 2.709(1.252～5.862), respectively.The AUC of FIB-4 was 0.799(0.704～0.895) for predicting inflammatory activity≥G2 or fibrosis stage≥S2 in HBeAg-negative patients with chronic HBV infected. FIB-4 also had good diagnostic performance for predicting antiviral treatment indications in patients of the validation group.The AUC of liver histology with Inflammatory activity≥G2 and Fibrosis stage≥S2 in patients with HBeAg negative chronic HBV infection was 0.821, higher than that of the group with only Inflammatory activity≥G2 or only Fibrosis stage≥S2. Conclusion FIB-4 can be used to determine the timing of antiviral treatment, which may reduce the need for liver biopsy of patients infected with HBeAg-negative chronic HBV infection.

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Objective To analyze the expression of tissue inhibitor of metalloproteinase-4(TIMP-4) and matrix metalloproteinase-2(MMP-2) in gastric cancer(GC) and adjacent tissues, and to explore their clinical significance in GC.Methods The expression levels of TIMP-4 protein and MMP-2 protein in 75 cases of gastric adenocarcino-ma tissues and 42 adjacent tissues were analyzed by immunohistochemical method, and the correlation betweenTIMP-4 protein and MMP-2 protein was analyzed by Pearson correlation analysis. Kaplan-meier survival analysis of GC in TCGA was performed to analyze the correlation between high TIMP-4 and MMP-2 expression in GC and overall survival. Results Among 75 GC tissues,65 cases were positive for TIMP-4 protein( 86. 67%),68 cases were positive for MMP-2 protein( 90. 67%),and 22 cases were positive for TIMP-4 in non-tumor tissues adjacent to cancer( 52. 38%),23 cases of MMP-2 positive in non-tumor tissues adjacent to cancer( 54. 76%). The differences were statistically significant compared with adjacent tissues( P<0. 01). The expression of MMP-2 protein was statistically related to lymph node metastasis( P<0. 001) and cancer stage( P<0. 05),but it was not statistically related to gender,age,tumor size or degree of differentiation. There was no statistical correlation between TIMP-4 protein expression,gender,age,tumor size,lymph node metastasis,tumor differentiation or cancer stage( P>0. 05). Pearson correlation analysis found a positive correlation between MMP-2 protein and TIMP-4 protein expression( r = 0. 351,P<0. 001). Kaplan-meier survival analysis showed that GC patients with high expression of TIMP-4 and MMP-2 had a poor prognosis. Conclusion MMP-2 protein and TIMP-4 protein are highly expressed in GC tissues,and the high expression suggest poor prognosis of GC patients,thus which may jointly participate in the occurrence and development of GC andwere expected to become a potential biological marker for predicting metastasis of GC.

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Objective To establish normal range of fetal anterior-posterior diameter, left-right diameter, and endometrium to lay the foundation for finding abnormal uterine development and diagnosing bisexual malformations. Methods The study subjects were 213 pregnant women with a gestational age of 21～41 weeks. Ultrasound was used to measure the anterior-posterior diameter, left-right diameter, and endometrium of the fetus.And analyzed the correlation between uterine development and gestational age. Results Sufficient measurements were obtained in 213 cases of fetal uterus examination. In the range of 21～34 gestational weeks,the relationship between fetal anterior-posterior diameter, left-right diameter, endometrium and gestational week was expressed by regression equation:anterior-posterior diameter(mm)=1.240×gestational weeks(r=0.887,P<0.0001)-23.969, left-right diameter(mm)=1.579×gestational weeks(r=0.915,P<0.001)-30.968,endometrium(mm)=0.802×gestational weeks(r=0.892,P<0.001)-16.585. Between the range of 35～41 gestational weeks,the 95% confidence interval of the population mean was calculated by usingtdistribution method:anterior-posterior diameter(15.992, 17.092) mm、left-right diameter(20.478, 21.627) mm、endometrium(8.805, 9.910) mm. There was a linear relationship between fetal anterior-posterior diameter, left-right diameter and endometrium. Conclusions The anterior-posterior diameter, left-right diameter and endometrium of the uterus increased with the increase of gestational age before 34 weeks, reached the slow growth stage to the plateau stage after 34 weeks, and no longer increased significantly with the increase of gestational age.

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The prokaryotic expression vector of Ltp1 was constructed by recombinant method, and transformed into E.coli BL21(DE3) for over-expression induced by isopropyl-β-D-galactoside(IPTG), then the target protein was coarsely purified by nickel column chromatography and fine purified by molecular sieve, and the product was expressed by SDS-PAGE electrophoresis and Western blot. The activity of the target protein was analyzed by color reaction with disodium p-nitrophenyl phosphate. SDS-PAGE results showed that there was an expression band consistent with its theoretical relative molecular weight. Western blot analysis confirmed that the protein was indeed a target protein with six histidine(His) tagswhich could catalyze the activity of phosphate monoester. The Michaelis constant(Km) and the maximum reaction rate(Vmax) were calculated to be 70.4 μmol and 51.1 μmol/(L·min) respectively, which had the highest catalytic activity at pH 5.0～6.0 and 37～50 ℃.

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A simple and efficient method of human ovarian serous adenocarcinoma cells(HOSACs), culture was established through improving methods for culturing primary cells of the common human ovarian serous adenocarcinoma. 25 tissue specimens of serous ovarian carcinoma confirmed by clinicopathology were collected, and each specimen was randomly divided into two groups: the modified group and the traditional group. The modified group was extracted cells with modified enzyme digestion method and tissue block adherence method(trypsin+hyaluronidase digestion for 2～3 min+tissue block and cell suspension plating). Traditional group was extracted with tissue block adherence method(the tissue block is shredded directly and then plating). The cell purity and cell viability obtained by the two methods were compared by cell immunofluorescence and CCK-8. The growth characteristics of primary cells of the two groups were observed under phase contrast microscope at 48 h and 72 h. The results of immunofluorescence and CCK-8 showed that the number of cells, cell viability and cell purity obtained in the modified group were higher than those in the traditional group(P<0.05).