Abstract:Objective：To study the effects of Pellino1’s post-translational modification—small ubiquitin-related modifier（SUMO）modification of Pellino1 in the nuclear factor κB（NF-κB）signaling pathway mediated by the TNF receptor associated factor6（TRAF6）. Methods：We constructed the plasmid of the SUMO modification sites mutantation of Pellino1，then cotransfected with TRAF6 plasmid and ubiquitin（Ub）plasmid into HEK-293 cells. The efficiency of plasmid transfection was observed by fluorescence microscopy. We detected the combination of mutant Pellino1 with TRAF6 by co-immunoprecipitation，and monitored the ubiquitination level of Pellino1 and TRAF6. Lipopolysaccharide（LPS）was used to induce inflammatory response，and the cytoplasmic nucleocapsid was extracted. Western blot was used to detect the phosphorylation of inhibitor of NF-κBα（IκBα）and the nuclear translocation of NF-κB P65. The effect of Pellino1 SUMO modification on TRAF6 mediated NF-κB signaling pathway was analyzed. Results：Compared with wild type Pellino1，mutant of Pellino1 SUMOylation ylation not only increased ubiquitination of Pellino1 itself，but also the combination with TRAF6，and the ubiquitination modification of TRAF6 were also increased. LPS stimulation significantly increased the phosphorylation of inhibitor of NF-κBα（IkBα）and NF-κB P65 nuclear translocation. High expression of mutant of Pellino1 SUMO after transfection significantly enhanced the LPS induced NF-κB signal activation. Conclusion：Mutant of Pellino1 SUMOylation can promote TRAF6 mediated activation of NF-κB signaling pathway by increasing the ubiquitination of Pellino1 and binding with TRAF6 and the ubiquitination level of TRAF6.

Abstract:Objective：To investigate the effect of melatonin on proliferation，migration and invasion of two rhabdomyosarcoma cell lines RD and RH30 and its mechanism. Methods：The proliferation，migration and invasion of both groups were detected by MTT，EdU and Transwell，respectively. Western blot was used to detect the expression of Gli1 in the downstream transcription factor of sonic hedgehog （Shh） before and after MT treatment. After inhibition and over-expression of Gli1 in the two groups，cell proliferation，migration and invasion were detected. Results：MT effectively inhibited the proliferation，migration and invasion of RH30 cells，but had no obvious effect on RD cells，and luzindole，the MT receptor inhibitor，did not change the inhibitory effect of MT on RH30 proliferation. The expression of Gli1 in RH30 cells decreased after MT treatment，and the cell proliferation，migration and invasion changed after Gli1 expression was changed. Conclusion：MT can significantly inhibit the proliferation，migration and invasion of RH30 cells，and its mechanism is related to the inhibition of Gli1 expression in RH30 cells by MT.

Abstract:Objective：To investigate the effects of ripasudil on proliferation and migration of human pulmonary arterial smooth cells（HPASMCs）induced by platelet-derived growth factor（PDGF）-BB and the mechanisms underlying. Methods：Cultured HPASMCs were divided into four groups：the control group，the PDGF-BB-treated group，the PDGF-BB and ripasudil-treated group and the ripasudil-treated group. CCK-8 was applied to investigate cell viability and EdU assay was used to evaluate the proliferation of HPASMCs. Transwell assay was employed to examine cell migration. The expression of matrix metalloproteinase-2（MMP-2）was determined by real-time PCR and Western blot. The levels of phosphorylated myosin phosphatase target subunit 1（MYPT1），extracellar regulated protein kinases 1/2（ERK1/2），p38，and protein kinase B（PKB/Akt）were detected by Western blot. Results：Compared with the control group，ripasudil blocked the proliferation and migration of HPASMCs challenged by PDGF-BB（P＜0.01）. Ripasudil suppressed PDGF-BB-induced upregulation of MMP-2（P＜0.05）. Moreover，ripasudil inhibited PDGF-BB-induced phosphorylation of MYPT1，ERK1/2，p38，and Akt（P＜0.05）. Conclusion：Ripasudil significantly inhibited PDGF-BB-induced proliferation and migration of HPASMCs，which might be attributed to the inhibition of MYPT1，ERK1/2，p38 and Akt. Ripasudil，a Rho kinase inhibitor，might be a potential therapeutic option in PAH.

Abstract:Objective：To investigate the effects of trophoblast cell-surface antigen 2（Trop2）on drug resistance of gastric cancer and the underlying mechanism. Methods：BGC823 cells were infected with the lentivirus encoding shRNA against Trop2 and puromycin was used to select stable cell lines；The effects of interference were detected by qRT-PCR，Western blot and immunofluorescence；CCK-8 proliferation test and flow cytometric detection were performed to detect the influence of daunorubicin on cell proliferation and apoptosis. The mRNA and protein expressions of TopoⅡα were detected by qRT-PCR and Western blot. Results：The mRNA and protein expressions of Trop2 in the shTrop2 group（transfected with Trop2 shRNA）were significantly lower than those in the control group（untreated）and the shNC group（transfected with normal control shRNA）；The shTrop2 group showed higher proliferation inhibition rates after daunorubicin treatment than these in the control group and the shNC group，and the IC50 value of the shTrop2 group was lower than that in the control group and the shNC group（P＜0.05）；The daunorubicin-induced cell apoptosis rate in the shTrop2 group was higher than that in the control group and the shNC group（P＜0.05）；The mRNA and protein expressions of drug resistance-related gene TopoⅡα decreased after interference with Trop2 expression stably. Conclusion：Trop2 can intensify daunorubicin resistance of gastric cancer cells by inhibiting the expression of TopoⅡα.

Abstract:Objective：To investigate the expression of calcium binding protein S100A16 in gastric cancer tissues and cells and its effect on the proliferation，migration and invasion of gastric cancer cell SGC-7901. Methods：Immunohistochemical staining was used to detect the differential expression of S100A16 in 8 pairs of gastric cancer and corresponding adjacent tissues. S100A16 overexpression plasmid and empty plasmid，interference plasmid and empty vector were transfected into human gastric cancer cell line SGC-7901 respectively，and the expression of S100A16 in each group was detected by Western blot. SRB staining and colony formation assay were used to detect the proliferation of cells in each group. Scratch test was used to detect the migration ability of cells in each group. Transwell assay was used to detect the invasive ability of each group. Results：The expression of S100A16 in gastric cancer tissues and gastric cancer cells were significantly higher than that in corresponding paracancerous tissues and normal gastric epithelial cells. SRB staining and colony formation assays showed that overexpression of S100A16 increased the proliferation of SGC-7901 cells and knocked down the proliferation of S100A16 cells. Scratch and Transwell experiments showed that overexpression of S100A16 increased SGC-7901 cell migration and invasiveness，while knockdown of S100A16 decreased. Western blot analysis showed that the expression of YBX-1 in SGC-7901 cells transfected with S100A16 over-expression plasmid was significantly increased compared with the control group. Co-immunoprecipitation experiments showed that S100A16 bound to YBX-1 in gastric cancer cell line SGC-7901. Conclusion：The expressions of S100A16 in gastric cancer tissues and cells are significantly up-regulated. The abnormal expression of S100A16 in gastric cancer may be due to the interaction with YBX-1，thereby promoting the proliferation，migration，invasion of gastric cancer cells and affecting the occurrence and development of gastric cancer.

Abstract:Objective：To investigate effect of HMG-CoA reductase degradation protein 1（HRD1） on the proliferation and migration of breast cancer cells. Methods：The expression of HRD1 and lactate dehydrogenase B（LDHB） was detected by Western blot in breast cancer tissue specimens and matched adjacent normal breast tissues；The expression of HRD1 and LDHB in MDA-MB-231 and MCF-7 cells transfected with adenovirus over-expressed HRD1 were detected by Western blot. Cell MTT assay，colony-formation assay，and transwell assay were performed to detect cell proliferation and cell migration rate，respectively. Results：Compared with the control group，the protein expression of HRD1 in matched adjacent normal breast tissues was increased，whereas，the protein expression of LDHB was decreased（P＜0.05）. A decline in cell proliferation and migration were found in MDA-MB-231 and MCF-7 cells transfected with adenovirus over-expressed HRD1（P＜0.05）. Conclusion：Overexpression of HRD1 results in the inhibition of growth and migration in human breast cancer cells. It downregulates the LDHB protein levels of human breast cancer cells.

Abstract:Objective：To detect the efficiency of c-Met CAR infection in T cells by qRT-PCR using specific primers. Methods：Gene recombination technology was introduced to construct c-Met CAR（GFP）retroviral plasmids. C-Met CAR and c-Met CAR（GFP）viruses were prepared by viral packaging. C-Met CAR-T cells and c-Met CAR-T（GFP）cells were prepared by infection of T cells with virus. The expression of c-Met CAR and c-Met CAR（GFP）virus infected 293T cells was tested by Western blot assay to express exogenous CD3ζ protein. Specific primers were designed to detect the infection efficiency of T cells by qRT-PCR and compared with flow cytometry. Results：c-Met CAR（GFP）plasmid was constructed，and the expression of c-Met CAR（GFP）plasmid on 293T cells was observed by fluorescence microscope. The results showed that c-Met CAR and c-Met CAR（GFP）virus infected 293T cells expressed exogenous CD3ζ protein. The infection efficiency of c-Met CAR and c-Met CAR（GFP）virus were（55.9 ± 2.3）% and（52.1 ± 1.7）% by qRT-PCR. The efficiency of c-Met CAR（GFP）infection was（50.7 ± 3.6）% by flow cytometry. There was no statistical difference between the two methods（P > 0.05）. Conclusion：Through the design of specific primers，the qRT-PCR can detect the infection efficiency of c-Met CAR virus on T cells，which is accurate and security，and have value for the clinical application of CAR-T cell therapy.

Abstract:Objective：To investigate the mechanism of endosomal sorting by constructing the transferrin receptor-ubiquitin（TfR-Ubi）protein. Methods：Mouse ubiquitin and human transferrin receptor genes were amplified to construct HA-TfR-Ubi plasmid. Then the expression of HA-TfR-Ubi protein was confirmed by Western blot following by transfected into A431 cells. The A431 and Hrs-KO MEF cells stably expressing HA-TfR-Ubi were treated with transferrin，and the location of HA-TfR-Ubi fusion protein was observed in the cells by immunofluorescence. Results：pHA-TfR-Ubi was successfully constructed. Immunofluorescence results showed that HA-TfR-Ubi fusion protein was sorted into late endosome /lysosomal，and its endosomal sorting was affected in Hrs-KO MEF cells. Conclusion：The endosomal sorting of TfR-Ubi fusion protein demonstrated that the ubiquitin label changed the transferrin receptor degradative pathway. TfR-Ubi plasmid provided a useful experimental tool for the study of ESCRT sorting mechanism.

Abstract:Objective：To investigate whether Del-1 is expressed in human periodontal ligament cells（hPDLCs），and the effect of lipopolysaccharides（LPS）on the expression of Del-1 in hPDLCs，and to explore the potential mechanism of Del-1 in the development of periodontitis. Methods：HPDLCs were isolated and cultured by tissue block method，and immunofluorescence assay，flow cytometry and Matrigel assay were used to identify cells. Immunofluorescence was used to detect if Del-1 was expressed in hPDLCs. Then variable concentrations of LPS were used to stimulate hPDLCs，and the expression of Del-1 was detected. The optimal concentration of LPS was selected，based on the most significant decrease of Del-1. The optimal concentration was selected to be used in the time-dependent experiment. The expressions of Del-1 and IL-6 were respectively assayed. hPDLCs were pretreated with Del-1 of variable concentrations（0，0.05，0.5 and 5 μg/mL）for 1 h，and then stimulated with LPS for 24 h. The expressions of IL-6 were assayed. hPDLCs were pretreated with the optimal concentration of Del-1 for 1 h，and then stimulated with LPS for 10 min，for detecting the phosphorylation of IκBα. Results：Del-1 could be expressed in hPDLCs，locating in cytoplasm of hPDLCs. The expressions of Del-1 were significantly down-regulated by LPS in a dose-independent and time-independent manner，which were contrary to IL-6. Del-1 could down-regulate the expression of IL-6 which was induced by LPS and inhibit the phosphorylation of IκBα in hPDLCs. Conclusions：Our study discovered for the first time that Del-1 could be expressed in hPDLCs，and it was down-regulated by LPS in hPDLCs，Del-1 could suppress the phosphorylation of NF-κB，playing an antagonistic role in the development of periodontitis.

Abstract:Objective：This study aimed at the effect of tumor necrosis factor receptor-associated factor 6（TRAF6）knockdown on interleukin-17（IL-17）modulating the expression of osteoprotegerin（OPG）and receptor activator for nuclear factor-κB ligand（RANKL）in human periodontal ligament cells（hPDLCs）under hypoxia. Methods：The experiment was set as the normoxia plus IL-17 group，the hypoxia plus IL-17 group and the hypoxia group. Each group was divided into the negative control group（the TRAF6-NC group）and the disturbance group（the TRAF6-shRNA group）. The shRNA lentiviral vector was used to infect hPDLCs. The efficiency of infection was observed by inverted microscope and verified by RT-PCR. The expression of HIF-1α，RANKL and OPG in human periodontal ligament cells were detected by Western blot and RT-PCR. Results：The ratio of RANKL/OPG relative expression of RANKL and OPG protein in the hypoxia plus IL-17 group was higher than that in the hypoxia group. Under hypoxia plus IL-17 conditions，the expression of RANKL was decreased in the TRAF6-shRNA group compared with the TRAF6-NC group，and the ratio of RANKL/OPG relative expression of RANKL and OPG was decreased. Conclusion：TRAF6 is involved in the regulation of IL-17-stimulated expression of bone resorption molecules in human periodontal ligament cells in hypoxic conditions. Knocking down TRAF6 can reduce IL-17-induced expression of bone resorption molecules in human periodontal ligament cells.

Abstract:Objective：To investigate the protective effects of adiponectin（APN）on apoptosis and extracellular matrix（ECM）degradation in rat nucleus pulposus（NP） cells under oxidative stress. Methods：Cell viability was detected by cell counting kit-8（CCK-8）to determine the optimal protective concentration of APN. Cells were randomly divided into the control group，the H2O2 group，the APN+H2O2 group and the compound C+APN+H2O2 group. Apoptosis incidence was evaluated by flow cytometry. The expression of Bcl-2，Bax，cleaved caspase-3，matrix metalloproteinase-13（MMP-13） and a disintegrin and metalloproteinase with thrombospondin motifs-5（ADAMTS-5）was detected by Western blot. The mRNA levels of collagen type Ⅱ alpha 1 chain（COL2A1） and aggrecan（ACAN） were detected by RT-PCR. The phosphorylation levels of adenosine 5′-monophosphate-activated protein kinase（AMPK）and mammalian target of rapamycin（mTOR） were evaluated by Western blot. Results：One μg/mL APN conferred the optimal protection against the cytotoxicity of 200 μmol/L H2O2. APN pretreatment significantly suppressed H2O2-induced apoptosis in NP cells. Consistently，the increase in Bax/Bcl-2 ratio and the expression of cleaved caspase-3 was also inhibited by APN. Although no notable inhibitory effect on ADAMTS-5 was observed，APN reduced the expression of MMP-13 induced by H2O2. Besides，the inhibition of H2O2 on the transcription of COL2A1 and ACAN was also notably abated by APN. APN also induced the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. Inhibition of AMPK with compound C alleviated the suppression of APN on mTOR phosphorylation. Moreover，the inhibition of APN on apoptosis and catabolism，together with the promotion of APN on anabolism were also reversed by compound C. Conclusion：APN inhibits H2O2-induced apoptosis and ECM degradation in rat NP cells in an AMPK/mTOR dependent manner.

Abstract:Objective：To evaluate the capacity of cartilage extracellular matrix-derived scaffolds prepared by a modified method for constructing tissue engineered cartilage. Methods：The cartilage pieces were fragmented using two different methods to produce extracellular matrix（ECM） powder and fine extracellular matrix（FECM） powder. ECM scaffold and FECM scaffold were fabricated with ECM powder and FECM powder through freeing and lyophilization technique. Both of ECM and FECM scaffolds were seeded with bone marrow mesenchymal stem cells（BMSCs）as the control group and the experimental group. The poly（lactic-co-glycolic acid）（PLGA） scaffold were seeded with BMSCs induced to differentiate into chondrocytes as the positive control group. After cultured in vitro for 12 h，the cell-scaffold constructs were implanted in the nude mice subcutaneously. The cell-scaffold constructs were harvested and detected on histology and immunohistochemistry at day 28. Results：Cell-scaffold constructs of the experimental group grew much better than that of the control group，and similar to the postive control group. More cartilage-like tissues were generated using FECM scaffold and stained strongly for sGAGs and type Ⅱ collagen. Conclusion：FECM scaffold can promote the differentiation of BMSCs into chondrocytes without exogenous transforming growth factor-β1（TGF-β1） and have potential of application in tissue engineering cartilage .

Abstract:Objective：To construct a sustained-release system to control the abnormally increase of chromium and cobalt ion concentration persistently and effectively in rats. Methods：First，a sustained-release system loaded with ethylene diamine tetraacetic acid （EDTA） was constructed. Then，the rat models were established by intra-articular injection of CoCrMo nanoparticles. Afterwards，the rat models were randomly divided into 4 groups，injected intraperitoneally with normal saline，EDTA，silica microsphere empty carrier solution and EDTA-loaded system solution，respectively，during which，the rats were injected with CoCrMo nanoparticles as before. The blood chromium and cobalt ion concentrations were measured. Results：After the therapy，the concentration of metal ions in groups receiving EDTA and EDTA-loaded system solution decreased significantly in the short run. However，as the time goes，the concentration of metal ions in group injected with EDTA increased and with EDTA-loaded system solution，stayed low，relatively，there was a statistically significant difference between these 2 groups. Conclusion：This sustained release system can effectively reduce the continuous increase of metal ions in the serum of rats，which can offer an alternative treatment for the abnormally increased metal ion after arthroplasties.

Abstract:Objective：To investigate the effect of astragaloside IV combined with enalapril on cardiac function after acute myocardial infarction（AMI）in mice. Methods：Twenty-four 8-week old male C57BL/6 mice were randomly divided into 4 groups，including the sham group，the CTL group，the enalapril group and the AS-IV+enalapril group. The CTL group，the enalapril group and the AS-IV+enalapril group were subjected to left anterior descending coronary artery ligation as AMI model. Mice in the enalapril group（enalapril 7.5 mg/kg）and the AS-IV+enalapril group（AS+IV 10 mg/kg + enalapril 7.5 mg/kg）were treated with the corresponding medicine，respectively. Echocardiography was applied to measure cardiac function 2 weeks later. Pathological changes of heart were studied by HE staining. CD31/ VEGFR3 double immunofluorescent staining were used to observe the density of vessels and angiogenesis. Results：Compared to the CTL group，the cardiac function was significantly improved in the enalapril group and the AS-IV+enalapril group，especially the latter one. Compared to the CTL group，there was significantly more angiogenesis to be observed in the peri- infarction region of the enalapril group and the AS-IV+enalapril group. This effect was better in the AS-IV+enalapril group. Conclusion：Astragaloside-IV combined with enalapril can effectively improve the cardiac function of mice after myocardial infarction and promote the regeneration of blood vessels in the peri-infarction region.

Abstract:Objective：To investigate the mechanism of miR-141 regulating cell proliferation in pancreatic cancer. Methods：Expression of miR-141 in normal pancreas cell and four pancreatic cancer cell lines were detected by quantitative reverse transcription PCR（qRT-PCR）. miR-141 expression file of GSE71533 which including 36 pancreatic cancer samples and 16 normal samples was downloaded from Gene Expression Omnibus（GEO）to analyze the expression of miR-141. Western blot was used to detect the expression of Bmi-1 in normal pancreatic cell and four pancreatic cancer cells Luciferase reporter assay was performed to analyze the relationship between Bmi-1 and miR-141. The effects of miR-141 and Bmi-1 on cell proliferation was examined by CCK8 and colony formation assay，respectively. Results：The qRT-PCR results showed that miR-141 was significantly down-regulated in pancreatic cancer cells. miR-141 was down-regulated in pancreatic cancer tissues by analyzing the microarray of GSE71533. Western blot analysis results demonstrated that Bmi-1 was up-regulated in pancreatic cancer cells. The dual luciferase assay indicated that miR-141 was anchored at the 3′-untranslated region of Bmi-1. Down-regulation of Bmi-1 inhibited the proliferation of pancreatic cancer cells. Upregulating miR-141 decreased the protein level of Bmi-1，thus repressing cell proliferation in pancreatic cancer cell. Conclusions：miR-141 acts as a tumor-suppressor in pancreatic cancer by targeting Bmi-1. These findings revealed that miR-141’s potential function in the treatment of pancreatic cancer.

Abstract:Objective： By adjusting the phosphorus content in adenine diet，we aimed to establish an effective chronic kidney disease mice model accompanied by hyperphosphatemia for further studies. Methods：Male C57BL/6 mice were divided into four groups：simple adenine dietary：the normal group（1.0% calcium，0.6% phosphorus），the simple adenine group（0.2% adenine，1.0% calcium，0.6% phosphorus），adenine integrating high phosphorus dietary：the normal group（0.8% calcium，0.6% phosphorus），the adenine integrating high phosphorus dietary group（0.2% adenine，0.6% calcium，1.0% phosphorus）（n=7）. The weight，the levels of blood urea nitrogen（BUN），calcium（Ca）and phosphorus（P）in serum were measured at 0 and 4 weeks after the start of adenine diet. The gene expression of fibrosis markers collagenⅠ，fibronectin（FN），plasminogen activator inhibitor-1（PAI-1），in-ammatory markers TNF-α，IL-1β and ICAM-1 in the kidney were detected by RT-PCR. Results：Compared to the control group，their bodyweights were decreased and the serum BUN level［（41.15 ± 4.59）mmol/L］was significantly increased at 4 weeks in the simple adenine group，while the levels of serum Ca［2.62 ± 0.16）mmol/L］and P［（2.22 ± 0.26）mmol/L］were slightly increased（P < 0.05）. In the adenine integrating high phosphorus dietary group，their weights were reduced and the level of serum P［（2.97 ± 0.29）mmol/L］showed significant increase，whereas the level of serum BUN［（14.68 ± 3.57）mmol/L］were slightly increased at 4 weeks（P < 0.05）. There was no significant difference in serum Ca levels. RT-PCR results showed that the expressions of collagenⅠ，FN，PAI-1，TNF-α，IL-1β and ICAM-1 were increased in two model groups. Conclusion：Adenine combined with high phosphorus diet feeding C57BL/6 mice for 4 weeks can establish a hyperphosphatemia model，accompanied by mild renal dysfunction and renal fibrosis，which is an effective method to set up the mice with hyperphosphatemia.

Abstract:Objective：To observe the negative effect of androgen on lactate production in human granulosa cells，meanwhile，to explore the follicular dysplasia in polycystic ovary syndrome （PCOS） with hyperandrogenism influenced by the reduced lactate. Methods：The human granulosa cell line-KGN cells were cultured in vitro and treated with different concentrations of testosterone（10-10-10-6 mol/L）for different time（0~48 h）. The content of lactate in culture supernatant was measured. The mRNA levels of LDHA and LDHB were quantified by real-time PCR. The protein expression of LDHA and LDHB were detected by Western blot. Results：After treated with 10-6，10-7 and 10-9 mol/L testosterone for 24 h，besides，10-8 and 10-9 mol/L testosterone for 24 h or 36 h，the contents of lactate in the supernatant were significantly decreased（P < 0.05）. The mRNA and protein expression level of LDHA and LDHB significantly decreased after treated with 10-7 or 10-9 mol/L testosterone for 24 h（P < 0.05）. Conclusion：Excess androgen can suppress the lactate production in human granulosa cell by inhibiting LDH，which may be related to follicular dysplasia in PCOS.

Abstract:Objective：To investigate the effect of inhibition of proinflammatory cytokines produced by macrophages on the progesterone resistance in endometriosis. Methods：The primary ectopic endometrial stromal cells（ESCs）of 15 endometriosis women were cultured. The chemotactic test was used to detect the role of CXCL12 expressed by ectopic ESCs in the recruitment of macrophages. LPS（100 ng/mL）was used to stimulate macrophages to express pro-inflammatory cytokines after macrophages were transfected with nuclear factor（NF）-κB decoy oligonucleotides（ODNs）. The electrophoretic mobility shift assay（EMSA）assay was used to detect the activation of NF-κB in macrophages after LPS stimulation，and ELISA was used to detect the expression of interleukin-1β（IL-1β）and tumor necrosis factor-α（TNF-α）in macrophages. In the co-culture system of ectopic ESCs and macrophages，the effect of NF-κB decoy ODNs on the decidualization of ectopic ESCs was observed. Results：CXCL12 secreted by ectopic ESCs played a role in the recruitment of macrophages in endometriosis. NF-κB decoy ODNs significantly inhibited the expression of pro-inflammatory cytokines in macrophages. In co-culture system of macrophages and ectopic ESCs，NF-κB decoy ODNs enhanced the progesterone-induced decidualization of ectopic ESCs by inhibiting the production of pro-inflammatory cytokines from macrophages. Conclusion：This study indicates that NF-κB targeting therapy of macrophages maybe an opportunity for mitigating the inflammatory response and progesterone resistance in endometriosis.

Abstract:Objective：To explore the effects of different doses of dexmedetomidine（DEX）preconditioning on focal in cerebral ischemia reperfusion injury rats. Methods：Fifty healthy male Sprague-Dawley rats were randomly divided into the sham operation group（sham group），the ischemia-reperfusion group（I/R group），the low-dose dexmedetomidine group（D1 group），the medium-dose dexmedetomidine group（D2 group），and the larger-dose dexmedetomidine group（D3 group）. The sham group received surgical procedures without causing ischemia. The sham group and the I/R group received 1 mL normal saline intraperitoneally 30 minutes before ischemia，and the D1，D2 and D3 groups were injected with dexmedetomidine in 50 μg/kg，100 μg/kg and 200 μg/kg，respectively. The levels of S100β protein were measured 24 h and 48 h after reperfusion，the contents of Bcl-2，Bax and the volume of cerebral infarction were determined 48 h after reperfusion. Results：Compared with the I/R group，the protein contents of S100β，Bax，and the volume of cerebral infarction in the D1，D2 and D3 groups were lower（P＜0.05），but were higher than those in the sham group（P＜0.05）；the Bcl-2 expression were increased（P＜0.05），but were lower than that in the sham group（P＜0.05）. The content of Bcl-2 was lower and the content of S100β protein at 24 h after surgery was higher in the D3 group than that in the D1 and D2 groups（P＜0.05），but were not statistically different between the D1 and D2 groups. The expression level of protein S100β，Bax and the volume of cerebral infarction were not statistically different between the D1，D2 and D3 groups. Conclusion：Dexmedetomidine preconditioning has neuroprotective effect on focal cerebral ischemia reperfusion injury in rats，and the mechanism may be related to the inhibition of apoptosis. The effect of dexmedetomidine has no significant difference between 50 μg/kg and 100 μg/kg.

Abstract:Objective：This project explored a random forest（RF） analysis of high-dimensional data with the confounding effects. Methods：We used computer simulations and real data validation to evaluate the performance of 2 methods which can potentially account for the confounding effects in RF analysis：RF analysis with maximum candidate variables at each split（RFMCV） and RF with glm-based correction. The distribution of ranks of the causal variable was used to evaluate these approaches. Results：Simulation experiments suggested that RF with glm-based correction was more effective than the RFMCV to correct the confounding effects. The real data validation showed that rs3754686 and rs2322660 were ranked first and second，respectively. Analysis results of GWAS data confirmed that RF with glm-based correction can effectively remove the spurious association between the LCT gene and height. Conclusion：The confounding effects should be correctly adjusted in RF analysis. RF with glm-based correction was applicable to adjust the confounding effects and variable selection in high-dimensional data.

Abstract:Objective：To detect the mutation of isoniazid resistance gene inhA-15（C-T） in drug-resistant tuberculosis strains by genechip technology，and to provide guidance for the treatment of drug-resistant tuberculosis patients. Method：From January 1，2014 to December 31，2016，patients with smear positive tuberculosis were continuously enrolled in TB designated hospitals in Lianyungang city of Jiangsu Province. The wild type and mutant type gene rpoB and wild type and mutant type of katG and inhA were detected by gene chip technique. Based on the results of traditional susceptibility test，the mutation frequency of related resistance loci，especially the mutation of isoniazid resistance related gene inhA-15（C-T），was analyzed in different drug-resistant strains. Results：A total of 1 356 patients with pulmonary tuberculosis were included in the final analysis. Through the detection of genechip technology，22（12.2%）strains had a single mutation at inhA gene mutation without katG mutation，Among them，3 cases of multidrug-resistant tuberculosis（MDR-TB） accounted for 15.4%，1 case of early extensive drug resistant tuberculosis（pre-XDR-TB） accounted for 7.7%，and 3 cases of extensively drug-resistant（XDR-TB） strains accounted for 23.1%. There were 32 cases（17.7%） had inhA gene mutation and / or katG gene mutation，of which 12（23.1%） were in MDR-TB，3（23.1%） were in pre-XDR-TB，and 3（23.1%） were in XDR-TB. Conclusion：More than 10% drug-resistant TB patients have a single mutation at the inhA-15 locus，indicating that large doses of isoniazid can be used to treat these drug-resistant patients. Although ethionamide（protionamide） had been widely used in the treatment of MDR-TB patients，almost 20% of drug resistant patients were resistant to these two drugs. The application of genechip has specifically reported gene mutations in drug-resistant tuberculosis patients，which will have some guiding effect on clinical treatment.

Abstract:Objective：To investigate the expression of HOXA9 protein in ovarian serous tumors and its potential as a specific marker for early diagnosis and drug resistance in ovarian cancer. Methods：Immunohistochemical staining for HOXA9 was performed in 178 patients with serous ovarian carcinoma，in 72 patients with ovarian borderline tumors and in 69 patients with benign ovarian tumors. The correlations between high HOXA9 expression and the clinicopathological features of the ovarian carcinomas were evaluated by the χ2 test and Fisher's exact test. The overall survival（OS）rates were calculated by the Kaplan-Meier method，and the association between prognostic factors and patient survival was analyzed by the Cox proportional hazard model. Results：The positive rate of HOXA9 protein in serous ovarian cancer（75.8%，135/178）was significantly higher than that in benign serous tumors（34.8%，24/69）（P < 0.05）. Similarly，the strong positive expression rate of HOXA9 protein in serous ovarian cancer（59.6%，106/178）was significantly higher than that in benign serous tumors（11.6%，8/69）（P < 0.05）. The positive expression rate of HOXA9 in high grade（65.1%，71/109）ovarian cancer was significantly higher than that in lower grade（47.8%，33/69）（P=0.014）. Similarly，the positive expression rate of HOXA9 in metastatic ovarian cancer（72.6%，69/95）was significantly higher than that in ovarian cancer without metastasis（42.2%，35/83）（P < 0.001）. Regarding the FIGO clinical stage，strong positive HOXA9 expression was 69.1%（65/94）in advanced（ⅡB-ⅢC）ovarian cancers and only 46.4%（39/84）in early（stageⅠ-ⅡA）ovarian cancers. The difference was statistically significant（P=0.003）. The OS rate of patients with strong expression of HOXA9 was lower than that of patients with weak expression of HOXA9（Log-rank=43.765，P < 0.001）. Multivariate analysis showed that HOXA9 was an independent prognostic factor in patients with ovarian cancer. Conclusion：HOXA9 expression is strongly associated with grade and outcome in ovarian carcinoma，and may serve as a useful molecular marker for clinical management.

Abstract:Colorectal cancer is a common digestive system disease with the increasing morbidity in the past 30 years. However，the pathogenesis of it remains unclear. Researches indicate that the occurrence of colorectal cancer is related to many factors such as heredity，diet，and inflammation. At present，chemopreventive drugs targeted at colorectal cancer show surprising results in clinical studies. These drugs are found to protect the body from chemical carcinogens and exert cancer prevention effect by promoting the expression of biphasic detoxification enzymes and antioxidant enzyme genes in the promotor region of nuclear factor E2 related factors. This article will review the latest researches in nuclear factor E2 related factor2 as a chemopreventive target in colorectal cancer.

Abstract:RNA-binding motif 4（RBM4） is an RNA-binding protein（RBP） that involves in different cell biological processes. Its structure contains two RNA recognition motifs and a CCHC-type zinc finger. RBM4 is expressed in a variety of tissues，at least in the process of post-transcriptional gene regulation，such as selective splicing of precursor mRNA，translational control and RNA silencing. RBM4 has a variety of biological functions，including maintaining embryonic and gonadal development，controlling circadian rhythms and mediating cell differentiation. Abnormal expression of RBM4 can lead to the occurrence of multiple diseases，especially in relation to tumors，but the specific mechanism needs further study. This article will review the research progress of RBM4.