Abstract:Objective：To construct a miRNA-205 overexpression vector and study the effect of miRNA-205 on the establishment of porcine induced pluripotent stem cell lines（iPSCs）. Methods：We compared miRNA transcriptional profiles between the primed porcine embryonic stem cells（primed PESCs），porcine inner cell mass（ICM）and pig fetal fibroblast cells（PEF）. The miRNA-205 precursor sequence was amplified by PCR and integrated into the expression vector pCAGDNA3-hFat1 to construct the overexpression vector pCAG-miR205. The expression vector was transfected into porcine fetal fibroblasts containing four mouse derived transcription factors（Oct4，Sox2，Klf4 and c-Myc）by lipofectamine transfection technique. To establish porcine iPSCs，the cells were incubated and cultured in stem cell culture medium. qPCR（real-time quantitative PCR）was used to detect the expression of miRNA-205 in the cells before and after transfection. The inducible effect of miRNA-205 on porcine iPSCs formation was analyzed. Results：The recombinant plasmid pCAG-miR205 was constructed. Compared with untransfected cells，the expression level of miRNA-205 in the transfected cells was significantly increased（P < 0.01），and the colony number was higher after being induced in stem cell culture medium. Conclusion：Over-expression of miRNA-205 at the cellular level can improve the efficiency of porcine induced pluripotent stem cell lineage. The miRNA-205 over-expression vector successfully constructed in this study will help to establish a stable cell line expressing miRNA-205，Which laid the foundation for further study on the exact function and mechanism of miRNA-205 on stem cell reprogramming and pluripotency maintenance.

Abstract:Objective：To investigate the expression and significance of MAP4K4 in pancreatic cancer（PC）and study its effect on proliferation. Methods：GEPIA and Oncomine databases were retrieved to analyze the expression of MAP4K4 in pancreatic tumors and normal pancreatic tissues；GEPIA and OncoLnc databases were performed to assess the relationship between MAP4K4 level and the survival time for patients with PC. Immunohistochemistry was used to detect the expression of MAP4K4 in human pancreatic tumors and adjacent normal tissues，then correlation and survival analysis were conducted to investigate the relationship between MAP4K4 level and clinicopathological parameters. qRT-PCR and Western blot were used to detect the expression of MAP4K4 in human PC cell lines. Lentiviruses were used to infect CFPAC-1 and MIA PaCa-2 to construct the cell lines with MAP4K4 overexpression and interfering，then CCK-8 and colony formation assay were conducted to study the influence of MAP4K4 on proliferation. Results：GEPIA and Oncomine databases showed that the expression of MAP4K4 was significantly higher in human pancreatic tumors than that in normal pancreatic tissues（P < 0.01）. GEPIA and OncoLnc databases revealed that higher expression of MAP4K4 was significantly associated with poorer survival time of PC patients（P < 0.01）. Data derived from immunohistochemistry indicated that MAP4K4 level was higher in human pancreatic tumors than that in adjacent normal tissues and was correlated with tumor size and differentiation（P < 0.05）. Survival analysis showed that the survival time of patients with high level of MAP4K4 was significantly lower than that with low level（P < 0.05）. Cell lines with MAP4K4 overexpression and interfering were successfully constructed. MAP4K4 overexpression significantly promoted，whereas MAP4K4 knockdown inhibited PC cell proliferaton compared with controls respectively（P < 0.05）. Conclusion：The expression of MAP4K4 is up-regulated in PC and correlated with poor prognosis，and it plays an important role in promoting proliferation in vitro.

Abstract:Objective：To explore the effect of TRIM22 on proliferation，invasion and migration in glioblastoma. Methods：The expression levels of TRIM22 were analyzed in TCGA database and were examined in two glioblastoma cell lines（U87 and U251 cells）using qRT-PCR and Western blot. Two independent TRIM22 siRNAs were transfected into U87 and U251 cells，and the effect of TRIM22 depletion was confirmed. CCK8 assay and colony formation assay were carried out to assess the effect of TRIM22 depletion on glioblastoma cell proliferation. Transwell assay and wound healing assay were also performed to investigate the role of TRIM22 depletion on glioblastoma cell invasion and migration. The expression of p-AKT and epithelial-mesenchymal transition（EMT）-associated proteins including N-cadherin，Vimentin，E-cadherin were examined using Western blot after TRIM22 depletion. Results：Inhibition of TRIM22 expression markedly reduced the proliferation，invasion and migration of glioblastoma cells. Correspondingly，the expression of p-AKT（S473），N-cadherin and Vimentin were decreased，whereas the expression of E-cadherin was increased after TRIM22 depletion. Conclusion：TRIM22 is a crucial factor for regulating the proliferation，invasion and migration of glioblastoma cells through PI3K/AKT signaling.

Abstract:Objective：To investigate the potential effects of long non-coding RNA（lncRNA）HOX transcript antisense intergenic RNA（HOTAIR）on proliferation，invasion and metastasis capacity of gastric cancer cells and to reveal the underling mechanisms. Methods：Human gastric epithelial cell line（GES-1）and different gastric cancer cell lines（SGC7901 and MKN-45）were cultured respectively. SGC7901 cancer cells were transiently transfected with small interfering RNA targeting HOTAIR（siHOTAIR），qRT-PCR was conducted to assess HOTAIR expression. MTT assay was carried out to evaluate the cell proliferation，wound-healing assay as well as transwell matrix invasion assay was performed respectively to evaluate the capability of invasion and migration of gastric cancer cells. The expression of cyclooxygenase-2（COX-2），together with E-cadherin was detected by Western blot assay. Results：The expression of HOTAIR in gastric cancer cells was higher than that in GES-1 cells（P < 0.05）. The cell proliferation was significantly inhibited after HOTAIR knockdown（P < 0.01）. Both wound healing assay and transwell assay demonstrated that HOTAIR knockdown resulted in a lower ratio in migration and invasion. Western blot analysis revealed that depletion of HOTAIR substantially decreased the expression of COX-2，and then elevated the expression of E-cadherin（P < 0.05），this effect would be attenuated by exogenous prostaglandin E2（PGE2）. Compared with siHOTAIR group，poliferation and invasion of SGC7901 cells were significantly increased in siHOTAIR + PGE2 group（P < 0.05）. Conclusion：Downregulation of lncRNA HOTAIR would inhibit cell viability，invasion and migration capability of gastric cancer cells，which were mediated partially by inhibiting the expression of COX-2 and then upregulating the expression of E-cadherin. HOTAIR might be a promising target to modulate the invasion and metastasis of gastric caner.

Abstract:Objective：To investigate the effects and mechanisms of 1，25（OH）2D3 on ameliorating dextran sulfate sodium（DSS）-induced colitis in mice. Methods：With DSS-induced colitis mice model established，inflammatory phenotype and goblet cells were detected and analyzed among model groups.The expression levels of tight junction proteins（TJs）among colonic tissues from different model groups were detected by RT-qPCR，Western blot and immunofluorescence staining. Using Caco-2 monolayer cell model，the protection of 1，25（OH）2D3 for TJs was detected in vitro. The mechanism of 1，25（OH）2D3 regulating expression of Claudin 4 was investigated via Chromatin immuno-precipitation（ChIP）and luciferase reporter assay. Results：1，25（OH）2D3 could enhance the intestinal epithelial barrier by promoting the expression of TJs，maintaining the structure of goblet cells and decreasing the intestinal permeability，thus relieving inflammation in DSS-induced acute colitis in mice. Conclusion：1，25（OH）2D3 plays a protective role in acute colitis mouse model via preserving intestinal permeability，promoting the expression levels of TJs and regulating the expression of Claudin4 through Vitamin Dreceptor.

Abstract:Objective：The aim of the study was to explore the effect and possible mechanism of 17 β-estradiol（E2）on the expression of small conductance Ca2+ activated K+ channel 3（SK3）in rat colonic smooth muscle.　Method：24 male Sprague-Dawley（SD）rats were randomly divided into 4 groups. Silicone tubes containing different solution was subcutaneously implanted in different groups of rats. Group C was filled with the solvent；Group EP was E2 in corn oil（0.3 mg/mL）which can keep serum levels of E2 at physiological levels；Group ES was E2 in corn oil（0.9 mg/mL）which can keep serum levels of E2 at supraphysiologic levels；Group EI was ICI 182780（estrogen receptor antagonist）plus E2（0.3 mg/mL）. The contraction of muscle strips and SK3 expression in colonic smooth muscles were detected at 14 days after tube implanted. SK3 were overexpressed in SMCs and Ca2+ concentration in cells were detected by laser scanning confocal microscope. Results：The contraction of muscle strips of EP group and ES group is weaker than other groups（P < 0.05）after Cch stimulating. And the SK3 expression was upregulated in EP and ES groups（P < 0.05）. Ca2+ decreased in SMCs after Cch stimulating in SK3 overexpression group and E2 stimulating group compared with control group. Conclusion：These findings indicate that E2 may suppress contraction of colonic smooth muscle by upregulating expression of SK3 and inhibiting Ca2+ influx in rats.

Abstract:Objective：To investigate the contribution of BDNF to exercise-induced angiogenesis in post-myocardial infarction rat hearts. Methods：Forty eight male SD rats（12 weeks old）were randomly divided into the following experimental groups：a sham infarction（Sham）group，a sedentary MI（MIC）group，a MI+exercise（MIE）group，a MI+exercise+K252a（MIEK）group. The MI model was established by ligation of the left anterior descending coronary artery，and exercised rats received 8 weeks of treadmill running. At the end of the exercise protocol，BDNF mRNA and protein expression（Western blot，RT-PCR and immunostaining）were measured，tropomyosin-related kinase B（TrkB）protein expression and phosphorylation levels were measured by Western blot，the angiogenic response was investigated by immunostaining，and cardiac function was analyzed by echocardiography. Results：Exercise training upregulated BDNF mRMA（P < 0.01）and protein（P < 0.01）expression，as well as phosphorylated TrkB levels（P < 0.01）in post-MI rat hearts. Exercise training induced elevation of serum BDNF in post-MI rats，which were related to angiogenesis（r=0.501，P=0.007）and cardiac function（r=0.611，P=0.002）. The exercise-related angiogenic response（P < 0.01）and functional improvement（P < 0.05）was attenuated by inhibition of BDNF receptor. Conclusion：Exercise convey its pro-angiogenic effects in ischemic myocardial tissue through BDNF.

Abstract:Objective：To evaluate the inhibitory effect of adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene on the xenograft tumor of hepatocellular carcinoma（HCC）in nude mice. Methods：A liver-specific adenovirus vector，Ad-CRM8-survivin-EHV4 TK（EHV4 TK），was constructed by molecular cloning and recombination. In nude mice bearing human hepatocellular carcinoma，the recombinant adenovirus was injected directly into the tumor followed by intraperitoneal injection of the prodrugs GCV and the inhibition effect on tumor xenografts was observed. The tumor section was stained with hematoxylin and eosin. Results： After adenovirus treatment, the volume of the tumors in EHV4 TK and GCV+EHV4 TK groups were（108.5 ± 18.3）and（37.6 ± 8.7）mm3. GCV+EHV4 TK represented much stronger anti-tumor effect on tumor growth than EHV4 TK only. GCV+EHV4 TK also showed much more tumor necrosis than EHV4 TK only. Conclusion：The adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene combined with GCV treatment may provide us a more effective，specific and convenient gene therapy method.

Abstract:Objective：To investigate the clinical efficacy of short-term intensive antiplatelet treatment of ticagrelor for patients with low response to clopidogrel after percutaneous coronary intervention（PCI）. Methods：A total of 100 cases who underwent PCI and were confirmed with low response to clopidogrel by light transmittance aggregation（LTA）were consecutively recruited and equally randomized into Clopidogrel（n=50）and Ticagrelor（n=50）groups. In Clopidogrel group，patients maintained clopidogrel 75 mg，gd in combination with aspirin 100 mg，gd；while in Ticagrelor group，patients were treated with ticagrelor 90 mg twice daily for 1 month，then switching to clopidogrel 75 mg，gd in combination with aspirin 100 mg，gd. The light transmission aggregations were determined for all patients 1 month after randomization；all participants were followed up and the adverse cardiovascular events were recorded for 6 months. Results：There were no significant differences between the two groups regarding both the adenosine diphosphate-induced platelet aggregation（PLADP）and the arachidonic acid-induced platelet aggregation（PLAA）prior to randomization（P > 0.05）. At 1 month after randomization，PLADP in the Ticagrelor group was significantly lower than that in the Clopidogrel group［（21.27 ± 12.81）% vs.（48.72 ± 10.92）%］（P < 0.01），while PLAA showed no significant difference between the two groups. The incidence of cardiogenic rehospitalization was significantly lower（6% vs. 20%，P < 0.05），although minimal bleeding was significantly higher（24% vs. 8%，P < 0.05）in the Ticagrelor group compared with that in the Clopidogrel group. Conclusion：The antiplatelet effect of ticagrelor is significantly more potent than that of clopidogrel，and 1-month intensive treatment of ticagrelor may reduce the cardiogenic rehospitalization in patients with CLR after PCI.

Abstract:Objective：To investigate the association of C-Reactive Protein（CRP）gene polymorphisms with low high-density lipoprotein cholesterol. Methods：Subjects were recruited by epidemiological cluster sampling in 2009 from Guanlin and Xushe town of Yixing city，and 4156 individuals were confirmed for final analysis. All subjects had completed physical examination and extracted of peripheral blood tests on an empty stomach blood lipids and other biochemical indicators in the morning. TaqMan probe method for four CRP gene SNPs genotyping. Multiple logistic regression was applied to further evaluate relationships between the SNPs and low-HDL-C. Results：The levels of hsCRP were significantly differet among rs1205 and rs876537 genotypes，P < 0.001. Rs876537 and rs1205 were significantly associated with decreased HDL-C，and the recessive model was statistically significant. The polymorphisms of rs1205 and rs876537 were significantly associated with low-HDL-C in the hs-CRP []1.0 mg/L group. Rs10737175 variation was significantly associated with low HDL-C in the hs-CRP normal group. Female with rs1205，rs10737175，rs876537 TT genotype were more likely to suffer from low-HDL-C. Female with rs10737175 TT genotype had a lower HDL-C level than CC+TT carriers. Conclusions：CRP polymorphisms were negatively associated with low-HDL-C level. The CRP gene rs1205，rs1073715 and rs876537 variations were associated with a significant reduction of HDL-C， and the association deserves more attention in the female population especially.

Abstract:Objective：To investigate the relationship of genetic variations in MAPK/ERK pathway with knee osteoarthritis risk. Methods：A case-control study was conducted including 278 patients with knee osteoarthritis and 289 age and sex matched healthy controls. A total of 5 potentially functional variations in MAPK/ERK pathyway（MEK1，MEK2，ERK1 and ERK2）selected by RegulomeDB were genotyped by using SequenomMassARRAY. Univariate and multivariate logistic regression models were used to evaluate the association and its strength. Results：In univariate analysis，rs350911 of MEK2 was significantly associated with knee osteoarthritis in recessive model（TT vs. TC+CC）（OR=2.62，95% CI：1.70~4.02，P < 0.01）. After adjustment for age，gender and BMI，the associations remain significant（OR=2.72，95% CI：1.75~4.22，P < 0.01）. The stratification analyses revealed that the effect of rs350911 on knee osteoarthritis was significant in male，lower BMI（<25 kg/m2），and lower Kellgren-Lawrence grade（1-2）（all P < 0.05）. P value for heterogeneity test was 0.01 in different gender group. Conclusion：The results indicate that potential functional genetic variation in MAPK/ERK plays an important role in the pathogenesis of knee osteoarthritis.

Abstract:Objective：To investigate the feasibility of multiphasic computed tomography scanning combined with a homemade software quantitative measurement of glomerular filtration rate（glomerular filtration rate measurement based on computed tomography，CT-GFR）. Methods：Fifty-two patients with unilateral renal tumors from February 2017 to July 2017 were included. All of them did not suffer from acute renal insufficiency. All patients underwent renal CT scans and serum creatinine（serum creatinine，Scr）tests preoperatively，of which 15 cases underwent additional radionuclide examination. Total CT-GFR and relative CT-GFR，with or without renal tumors，were calculated by a homemade GFR quantitative software. The correlation between total CT-GFR and estimated glomerular filtration rate（eGFR）was analyzed by Pearson’s method. The correlation between relative CT-GFR and relative GFR measurement based on radionuclide examination（R-GFR）of split kidneys was also analyzed by Pearson’s method. Total CT-GFR and relative CT-GFR were calculated in early and late renal parenchymal phases respectively with or without renal tumors. Results：Total CT-GFR in early renal parenchymal phase was highly correlated with eGFR（r=0.81，P < 0.01）. Relative CT-GFR in early renal parenchymal phase was highly correlated with relative R-GFR in the split kidneys（r=0.85，P < 0.01）. Conclusion：Split and total GFR can be measured accurately using multiphasic CT combined with homemade software in patients with unilateral renal tumors.

Abstract:Objective：To study the effect of enhanced recovery after surgery（ERAS）concept in perioperative period of laparoscopic radical prostatectomy. Methods：Total 91 cases with prostatic cancer which underwent operations in our hospital from December 2015 to September 2017 were divided into two groups，control group and ERAS group. All patients of two groups accepted laparoscopic radical prostatectomy. The data of recovery of gastrointestinal function，catheter and drainage tube removal time，length of stay（LOS），hospitalization expenses，incidence of postoperative complications and recovery rate of urinary incontinence 3 months after discharge were compared between the two groups. Results：Compared with control group，cases in ERAS group were with shorter time of recovery of gastrointestinal function（P < 0.05），shorter LOS（P < 0.05），shorter catheter and drainage tube removal time，lower hospitalization expenses and higher recovery rate of urinary incontinence（P < 0.05）. Conclusion：Methods of ERAS used in perioperative period of laparoscopic radical prostatectomy can apparently accelerate the time of postoperative recovery，shorten time stay in hospital and cut down the total cost，finally improve patients’ satisfaction.

Abstract:Objective：To investigate the effect of miR-448 on the proliferation of oral squamous cell carcinoma，and to screen and verify the relvelant genes. Methods：Using RT-qPCR to study the expression of miR-448 in human OSCC. Then miR-448 inhibitor was transfected into OSCC cell line Cal-27 cells，and the effects of miR-448 inhibitor on proliferation and migration of OSCC were detected by MTT assay and scratch test. In addition，we screened the target gene of miR-448 by three gene prediction software，and verify the regulation of miR-448 on target genes. Results：We found that miR-448 was significantly increased in 15 pairs of OSCC tissues by RT-qPCR analysis. Compared with the control group，the proliferation and migration of Cal-27 cells were significantly decreased after transfected with miR-448 inhibitor. The expression of SPARCL1 gene was increased detected by RT-qPCR and Western blot，and the presence of site-specific binding of miR-448 to the SPARCL1 gene in the 3′ UTR region was confirmed by luciferase reporter. Conclusions：MiR-448 is highly expressed in clinical samples of oral squamous cell carcinoma. miR-448 down-regulates the expression of SPARCL1 by binding to SPARCL1-3′ UTR，playing an oncogene role in oral squamous cell carcinoma and promoting tumor cell proliferation and migration.

Abstract:Objective：To investigate the effect of bioactive composite layers on porous titanium（Ti）surfaces with dopamine adhering hydroxyapatite coating on rat bone marrow mesenchymal stem cells. Methods： Coating the Ti with dopamine，the modified surface to increase the hydroxyapatite（HA）’s adhesion onto the titanium implant surface which has porous structure treating with the alkali-heat to form bioactive composite layers. Scanning electron microscopy（SEM）and energy dispersive X-ray spectroscopy（EDX）were employed to investigate the surface evolution of titanium after the chemical surface treatment and HA adhesion. Bone marrow deriued stem cells（BMSCs） were cultured and initial cell attachment，morphology，proliferation were evaluated. Results：The bioactive composite layers on porous titanium surfaces with dopamine adhering hydroxyapatite coating was successfully prepared. The bioactive composite layers were applied in vitro，the growth of BMSCs was not inhibited on it. Conclusion：The bioactive composite layers significantly promote the adhesion and early proliferation of BMSCs in vitro. This finding shows that the proposed method demonstrates a good prospective application in surface modification of titanium.

Abstract:Objective：To investigate the root anatomy and canal configuration of the mandibular first premolars with radicular groove in root surface by micro-computer tomography（Micro-CT）. Methods：In total of 58 mandibular first premolars with radicular groove were selected. The 3-dimensional images of the teeth and canal system morphology were reconstructed by Micro-CT. The types of canal configuration were classified，and the position and angle of Vertucci V root canal bifurcation were measured. The location and incidence of the accessory canals were recorded. The location of the apical foramen，the wall thickness and the diameter of the root canal were analyzed. Results：The most common type of root canal in mandibular first premolar with radicular groove was type V canal. In this study，cases of type V accounted for 55.2%. Root canal furcation of type V mandibular first premolar located about 6.13 mm below the cemento-enamel junction（CEJ）. Significant differences were found in the apical position，wall thickness and root canal diameter between the buccal and lingual canal. Conclusion：The root canal morphology of mandibular first premolars with radicular groove was complex，with a high incidence of type V canals. Understanding the anatomic features of the root canals provides greatly help in clinical treatment.

Abstract:Objective：To explore the mechanism of the chronic stress contribution to periodontitis. Method：Sprague-Dawley rats（2 months）were received chronic unpredicted mild stress（CUS）for eight weeks. Then，four weeks later，they were devided into susceptible（depressive）or unsusceptible（resistant）groups according to sucrose preference test. Total body and adrenal weights of rats were weighed weekly and at 8 week，respectively. The levels of plasma corticosterone and insulin，the total insulin receptor（IRS）and activity-dependent phosphorylated IRS in the periodontal tissues，and the mRNA expression of IL-1β and TNF-α in the periodontal tissues were detected by enzyme-linked immunosorbent assay（ELISA），Western blot and Real-time PCR，respectively. Results：After 4 weeks of CUS，59% of rats showed a decrease of sucrose preference indicating anhedonic or depressive-like behavior（CMSA）. Compared with controls，all the stressed rats showed decreased weight after three weeks；after eight weeks，they also exhibted increased corticosterone and decreased insulin in plasma，decreased p-IRS and increased mRNA levels of IL-1β and TNF-α in the periodontal tissue. The depressive andresitant groups showed no significant differences in above measures. Conclusion：The stress itself，rather than stress-induced mood disorders，leads to HPA axis disorders，abnormality of insulin signal pathway，and increased expression of periodontal tissue inflammatory factor. These changes may serve as the mechanism in contibuting to stress susceptible periodontal disease.

Abstract:Myelodysplastic syndrome（MDS）is a group of myeloid stem cell clonal disorders，characterized by a various degree decrease of peripheral blood cells，ineffective and pathological bone marrow hematopoiesis. There is large variability in the risk of progression to acute myeloid leukemia（AML）. The pathogenesis of the disease become clear gradually：the imbalance of the immune system，especially cellular immune defects，lead to the abnormal cloning clear blocked，abnormal precursor cells immune escape，while a variety of secretion cytokines promote apoptosis of bone marrow cells early. In this article，the changes of natural killer cells，lymphocyte，dendritic cells and marrow-derived suppressor cells of myelodysplastic syndrome and acute myeloid leukemia are reviewed in recent years in order to optimize the monitoring MDS progress.

Abstract:Phosphoinositol 3-kinase（PI3K）/ serine threonine protein kinase（Akt）signaling pathway is one of the important signaling pathways in the body，which can regulate the biological activity of cell growth，proliferation and apoptosis. A large number of studies have shown that activation of PI3K / Akt signaling pathway is involved in inflammatory responses and regulation of immune responses. Akt is a key downstream protein of the PI3K signaling pathway. Therefore，the mechanism of Akt downstream protein on the regulation of inflammatory diseases such as shock，sepsis and ischemia-reperfusion injury is helpful to find a potential target for the treatment of these diseases.