Objective:To study the expression of miR-496 in gastric cancer cells, and explore its role and mechanism in the proliferation and apoptosis of gastric cancer cells. Methods:Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression of miR-496 in normal gastric epithelial cell lines and gastric cancer cell lines AGS and MKN45. miR-496 was knocked down in AGS cells with the lowest expression level, and a negative control group and a blank control group were set up. Cell proliferation and apoptosis were detected by CCK8 assay and flow cytometry. LYN, the target gene of miR-496, was screened using bioinformatics software, and the effect of transfection of miR-496 on LYN expression was detected by qPCR. Subsequently, rescure experiment was conducted to further study the mechanism of miR-496 on gastric cancer cells through regulation of LYN. Data were analyzed by GraphPad Prism 9 software. Measurement data were presented as mean ± standard deviation, and the comparison between the two groups was performed by t test. Results:The expression of miR-496 in AGS and MKN45 was significantly lower than that in normal gastric epithelial cells ( P<0.05). After overexpression of miR-496, the proliferation of AGS cells could be inhibited and the apoptosis ratio of AGS cells could be significantly increased ( P<0.05). QPCR results showed that miR-496 overexpression group could inhibit the expression of LYN ( P<0.05). Bioinformatics analysis showed that miR-496 binds to LYN kinase ( LYN) 3 ′UTR region, and overexpression of miR-496 can inhibit the expression of LYN in AGS cells, while CCK8 rescue experiment showed that overexpression of LYN could remove the inhibitory effect of miR-496 on cell proliferation. Flow cytometry showed that LYN expression could cancel the promoting effect of miR-496 on apoptosis ( P<0.05). Conclusion:miR-496 is low expressed in gastric cancer cells, and it inhibits the proliferation and promotes apoptosis of gastric cancer cells by targeting the expression of LYN in gastric cancer cells.

Objective:To explore the features the gait of elderly persons with type 2 diabetes and peri-pheral neuropathy.Methods:Twenty patients no less than 60 years old with type 2 diabetes and peripheral neuropathy (DPN) formed a DPN group, while 20 counterparts with type 2 diabetes but without peripheral neuropathy composed the DM group, and another 20 healthy counterparts served as a control group. The three groups were tested using the Swedish Qualisys motion capture system and their walking speed, step length, step width, stride frequency and stride length, bipedal foot support phase time, single foot support phase time, peak plantar pressure, and regional-holding time were collected and compared.Results:The average walking speed, stride length and stepping frequency of the DPN group were all significantly lower than the other 2 groups′ averages. Their bipedal support phase was significantly longer, but their single foot support phase time was significantly shorter. And in the DPN group the average first and second peak plantar pressures and the second peak pressure time were significantly greater than the other groups′ averages.Conclusions:Elderly patients with type 2 diabetes and peripheral neuropathy have significant gait abnormalities, decreased walking stability, as well as increased plantar pressure and plantar compression time.

Objective:To study the effect and mechanism of Rab4A knockout expression on proliferation, migration and invasion of gastric cancer cells. Methods:The expression of Rab4A in four human gastric cancer cell lines MGC-803, SGC-790, MKN45 and AGS was detected by Western blot. Rab4A was knocked out in AGS cells with the highest expression level, and untransfected gastric cancer cells were used as control group. Cell proliferation, migration and invasion were detected by CCK8 and Transwell assay, respectively. Western blot analysis was used to investigate the expression changes of epidermal growth factor receptor (EGFR), downstream pathway proteins AKT and β-catenin induced by Rab4A knockout. The interaction between Rab4A and MiR- 496 was detected by dual luciferase reporter gene, and the effect of MiR- 496 transfection on Rab4A expression was detected by qPCR and Western blot. GraphPad Prism 9 software was used for data analysis, t-test was used for comparison between the two groups, and normal distribution measurement data were expressed as mean ± standard deviation ( ± s). Results:The expression of Rab4A was the highest in AGS cells, and the knockdown of Rab4A inhibited the proliferation, migration and invasion of AGS cells ( P<0.05). In Rab4A knockout gastric cancer cells, the surface expression of epidermal growth factor receptor (EGFR) was significantly decreased, and the expression of downstream pathway proteins p-AKT and β-catenin was also inhibited ( P<0.05). The luciferase reporter showed that MiR- 496 could bind the 3′UTR of Rab4A. In addition, MiR- 496 down-regulated the expression of Rab4A in AGS cells( P<0.05). Conclusion:The expression of Rab4A is inhibited by MiR- 496, and the proliferation, migration and invasion of gastric cancer cells can be inhibited by down-regulating the surface expression of EGFR after inhibiting Rab4A expression.

Objective To evaluate the predicting value of circulating miRNAs for sepsis secondary to pneumonia in elderly patients.Methods From April 2016 to January 2017,44 cases with sepsis secondary to pneumonia,52 elderly patients with pneumonia and 21 healthy older adults as control were involved in this study.The expression levels of MiRNA-150 5p,miRNA-25-3p,miRNA-122 5p and miRNA-223-3p in plasma were evaluated by fluorescence quantitative PCR.The demographic characteristics,sequential organ failure assessment (SOFA)scores,prognosis and days stayed in ICU were recorded.The area under the receiver operating charaeteristic(ROC)curve was used to calculated the specificity and sensitivity of miRNA in identifying sepsis-associated pneumonia.Results There were significantly differences among levels of circulating miRNA-223-3p in pneumonia,sepsis and healthy control groups(F =36.441,P =0.000),△CT values were 2.39 ± 1.36,1.44± 1.43,and 4.58 ± 0.91,respectively.The relative expression levels of miRNA-223-3p in the three groups were significantly different (P =0.000),which were 0.189 (0.107,0.367),0.361 (0.221,0.735),and 0.044 (0.022,0.061),respectively.The AUC of miRNA-223-3p for predicting sepsis from pneumonia was 0.964(95 ％CI =0.925 1.000).At a cutoff value of 2.759,miRNA-223-3p yielded a sensitivity of 82.9％ and a specificity of 100.0％.Conclusions MiRNA-223-3p expression is up-regulated in patients with sepsis secondary to pneumonia compared to that of patients with pneumonia,and it could be used to predict sepsis associated pneumonia.

Objective To construct the lentiviral vector of RNA interference(RNAi) for DEK,and to detect its effect on breast cancer cell growth.Methods The DEK siRNA was designed and constructed based on DEK sequence using a lentiviral vector.The lentivirul vector containing DEK siRNA was named PSIH-H1-DEK as confirmed by PCR and sequenceing.PSIH-H1-DEK was then packaged with accessory plasmids into lentivirus in 293T cells and selected for 2 weeks with puromycin ( puro ) before the mixed colonies stably expressing DEK siRNA were obtained and the DEK expression was detected by real time PCR( RT-PCR) and Western blotting.The effect of DEK siRNA on ZR75-1 cell growth was determined by cell counting kit.Results Western blot and RT-PCR showed that PSIH-H1-DEK siRNA could suppress DEK gene expression.Suppression of DEK could markedly inhibit the growth of ZR75-1 cells.Conclusion The lentivirus-mediated DEK siRNA is obtained,which will facilitate further research on DEK function in breast cancer development.

Objective To construct the lentiviral vector (pSIH-H1) for E6AP small-interfering RNA(siRNA) and to detect its effect on breast cancer ZR 75-1cell growth.Methods E6AP siRNA was designed and constructed based on hu-man papillomavirus E6-associated protein ( E6AP) cDNA sequence.The expression of E6AP was examined by real-time quantitative PCR(qRT-PCR) and Western blotting.The effect of E6AP on ZR75-1 cell growth was determined by cck-8 kit.Results DNA sequencing indicated that E 6AP siRNA expression vector was constructed successfully .qRT-PCR and Western blotting experiments showed that pSIH-H1-E6AP siRNA could suppress the E6AP gene expression.Suppression of E6AP could markedly inhibit the growth of ZR 75-1.Conclusion A lentivirus RNA interference ( RNAi) vector targeting E6AP gene is successfully constructed ,which inhibits the cell growth of ZR 75-1.

ObjectiveTo assess the effect of transarterial radioembolization (TARE) using Yttrium-90 microsphere for unresectable primary liver cancer (PLC).MethodsLiterature from January 2009 to December 2013 were searched in the Medline, Web of Science, Cochrane Controlled Trial Register (CENTRAL) and EMBASE databases with the search terms mainly including: radioembolization, transarterial radioembolization, TARE, selective internal radiation therapy, SIRT, Yttrium-90, 90Y, chemoembolization, transarterial chemoembolization (TACE), hepatocellular carcinoma, HCC, liver cancer, liver tumor, liver neoplasm and with the assistance of manual searching. Data of the included literature were merged and the patients were divided into TARE group and TACE group according to the different treatments. The data of tumor therapeutic response and 1-, 2-, 3-year survival rates were collected. Literature heterogeneity inspection was conducted byQ test. Publication bias was tested by drawing funnel plot and linear regression model.ResultsFive articles were included after screening with the quality of medium to high. There were totally 591 cases with 292 in TARE group and 299 in TACE group. Meta-analysis was conducted using fixed effect model. Tumor therapeutic response was observed better in TARE group, compared with that in TACE group (RR=1.50,P<0.05). The 2-, 3-year survival rates in TARE group were signiifcantly higher than those in TACE group (RR=1.56, 2.04;P<0.05).ConclusionsCompared with TACE, TARE can obviously improve the tumor therapeutic response rate and long-term survival rate of patients with unresectable PLC.

Objective To compare the results of the anterior approach (AA) with the conventional approach in the treatment of large hepatocellular carcinoma (HCC).Methods We searched the Medline,PubMed,Cochrane Library,Wanfang database on randomized clinical controlled trials and non-randomized clinical controlled trials comparing AA with the CA in right hepatic resection for large hepatocellular carcinoma.The data were analyzed with the RevMan5 software.Results Five non-randomized clinical controlled trials (NRCTs) and three randomized clinical controlled trials involving 615 patients (304 in the AA group,311 in the CA group) were enrolled into the analysis.There was no significant difference in the operation time between the two groups.Compared with the CA,the AA had lower intraoperative blood loss (WMD=-680.2 ml; 95％CI,-1023.97～-336.43;P=0.0001),blood transfusion rate (OR=0.38;95％ CI,0.25～0.59;P＜0.0001),intraoperative tumor rupture (OR=0.33;95％CI,0.11～0.97;P=0.04),surgical complication (OR=0.59;95％CI,0.38 ～ 0.93 ; P =0.02),hospital mortality (OR =0.37 ; 95 ％ CI,0.21 ～ 0.67 ; P =0.0009),and hospital stay (WMD=-4.75 d;95％CI,-7.82～-1.67;P=0.002).Conclusion AA is superior to CA in the treatment of larger.The operation time is the same for the 2 approaches.

Objective To research a new type of gastrointestinal anastomat -- anastomosis device with three rows of staples (Pa-tent No .2012200583213) ,and clarify its associated functions through esophagogastric anastomosis operations on pigs ,so that we could provide experiences for its clinical application in future .Methods Compared with domestic anastomat with two rows of sta-ples ,we designed and manufactured a new type of anastomat with three rows of staples and researched its function .Choosing 12 do-mestic pigs ,about 60 kg ,gastroesophageal anastomosis was taken twice with each case by anastomat with three or two rows of sta -ples randomly in sequence .According to the different types of anastomats ,cases were divided into two groups :group A ,used anas-tomat with three rows of staples ,including 12 cases of anastomosis ;group B ,used anastomat with two rows of staples ,including an-other 12 cases of anastomosis .Results Compared with group B ,cases of group A have less bleeding sites (t = 7 .00 ,P < 0 .01) . Without reinforcement and with 0 .5 kg of tension ,fewer of outermost staples exposed(t= 6 .17 ,P< 0 .01) .And the shape of used staples of group A is double circles ,which has bigger mechanical strength than that of group B (t= 6 .57 ,P < 0 .01) .Conclusion The function of anastomat with three rows of staples surpasses that of traditional anastomat with two rows of staples in pig esopha -gogastrostomy surgery .

OBJECTIVETo study the constituents of the whole herbs of Vernonia cinerea by bio-activity guided isolation with PC-12 model.

METHODThe constituents were separated by column chromatography and the structures were elucidated by spectroscopic methods.

RESULTTen compounds were identified to be (-)-clovane-2,9-diol (1), caryolane-1,9beta-diol (2), apigenin (3), chrysoeriol (4), luteolin (5), thermopsoside (6), luteolin-7-O-beta-D-glucoside (7), quercetin(8), apigenin-4'-O-beta-D-glucoside (9), hyperin (10), beta-amyrin aceate (11), lupeol acetate (12).

CONCLUSIONCompounds 1, 2, 6 and 10 were isolated from this genus for the first time.