Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Aim To construct a subtracted cDNA library o f differentially expressed genes in human gastric carcinoma induced by diallyl dis ulfide(DADS). Methods Differentially expressed cDNA species induc ed by DADS in MGC 803 human gastric carcinoma cell line was determined by using suppression subtractive hybridization (SSH). Then these cDNA species were direct ly inserted into T/A cloning vector to set up the subtractive library. Amplification of th e library was carried out with transformation of E.coli by high voltage electrop erforation. One hundred positive bacteria clones were randomly picked and identi fied using PCR method. Results The amplified library contained more than 1,000 positive bacteria clones. Random analysis of 100 clones with PCR m ethod showed that all clones contained 100～600 bp inserts.Conclusions A subtracted cDNA library of differentially expressed genes in MGC 803 hum an gastric carcinoma cell line induced by DADS is constructed successfully with SSH and T/A cloning techniques. The library is efficient and lays solid foundati on for screening and cloning new and specific tumor correlative genes of human g astric carcinoma, and provides a new idea for further exploring the mechanism of DADS effects on carcinoma cells.

Aim To investigate whether DATS induce MGC803 cell apoptosis and the relationship betweenapoptosis and Ca~(2+) disruption. Methods MGC-803 cell growth inhibition was measured by MTT assay. Tunnel and flow cytometry methods were used to determine the induction of apoptosis and Ca2+ homeostasis disruption. Result MTT assay showed that the inhibitory rates on MGC-803 cell growth of different concentrations of DATS 4,8,12,16 and 24 mg?L-1 were 0.231?0.037,0.305?0.036,0.455?0.029,0.607?0.058,0.751?0.019 respectively. Flow cytometry analysis showed that treating MGC803 cell with DATS significantly increased the percentage of apoptosis cells and intracellular Ca2+. Treatment of cells with 1,2-bis(2-aminophenoxye-thane)-N,N,N-tetraacetic acid tetrakis acetoxymethyl ester (BAPTA-AM), cellular Ca2+ chelator, resulted in abolishment of the elevation of intracellular Ca~(2+) and blockage of DATS induced apoptotic of MGC-803. Conclusoin DATS could induce apoptosis of MGC-803 cells through the mechanism of Ca~(2+) homeostasis disruption.

Aim To observe effect of DADS on the differentiation of gastric adenocarcinoma cells and acetylation of human gastric cancer tumor cell transplantable histone.Methods Human gastric adenocarcinoma heterotransplantable tumor model was constructed through subcutaneously injecting MGC803 cells to nude mice.Morphologic changes of xenograft tumor cells were observed with optical microscope,the influence of DADS on xenograft tumor cells generationcycle distribution,the expression of p21~(WAF1) protein,histone H3 and and H4 acetylion were analyzed with flow cytometry and Western blot.Results There was obvious growth inhibitory effect of xenograft tumor while abdominal injection dose were 100 and 200 mg?kg~(-1)DADS;cells density and heteromorphism decreased after treated with DADS.Flow cytometry analysis revealed that treating xenograft tumor cells with increasing quantities of DADS increased the percentage of cells in the G_2/M phase.The proportion of xenograft tumor cells in the G_2/M phase after treatment with 100 mg?kg~(-1) and 200 mg?kg~(-1) DADS was 2.22 and 3.37 times of that in NS group.Western blot analysis showed H3 acetylion increase along with G_2/M arrest of xenograft tumor cells by DADS.DADS didn′t influence the expression level of H4 acetylion;the expression of p21~(WAF1) protein in xenograft tumor increased along with the increases in the concentration of DADS.Conclusion DADS cansignificantly inhibit the growth of human gastric carcinoma xenograft in BALB/C nucle mice and induce cell differentiation,which might be related with up-regulation of histone a cetylization and p21~(WAF1) protein level.

Objective To investigate the efficacy and side effects of the molecular targeted drug sunitinib in the treatment of metastatic renal cell carcinoma(mRCC). Methods Fifteen patients with histopathological confirmed mRCC,including 10 males and 5 females,were enrolled in the study.The median age was 56(range from 37 to 73 years).Fourteen cases of clear-cell RCC and 1 papillary RCC were diagnosed.Thirteen patients had prior nephrectomy,and 6 patients were treated with cytokines previously.All of the patients were given sunitinib at a dosage of either 50 mg daily(4 weeks on/2 weeks off)or 37.5 mg daily continuously.Objective response rate(ORR),progressive-free survival(PFS)and overall survival(OS)were evaluated,and adverse events were also observed. Results The median follow-up was 13 months(2-24 months).According to RECIST,partial response could be evaluated in 8 patients and stable was found in another 5 patients.Only 2 patients were confirmed disease progression.The ORR was 53% with the disease control rate of 87%,However,the median PFS and OS were not yet available due to the short-term follow-up.During the treatment,the most common adverse events experienced by patients were hand-foot syndrome 11/15(73%),changes in hair color 10/15(67%),mucositis 9/15(60%),alopecie 9/15(60%),diarrhea 8/15(53%)and neutropenia 8/15 (53%). All of adverse events were manageable and reversible. Conclusions Sunitinib as a single agent in the treatment for Chinese mRCC patient is efficient and the side effects are reversible. Further long-term follow-up and expanded samples should be expected to confirm the efficacy and safety of sunitinib.

Objective To investigate the correlation and consistency between the parameters of thromboelastography(TEG)and routine coagulation tests,and to evaluate the application value of the two methods in heparin anticoagulation monitoring and coagulation function monitoring in patients receiving extracorporeal membrane oxygenation(ECMO)therapy.Methods A total of 138 patients who recieved ECMO in the Department of Critical Care Medicine of the People's Hospital of Guangxi Zhuang Autonomous Region from October 2021 to December 2022 were selected.A total of 317 pairs of ordinary TEG and heparinase-modified TEG(hmTEG)parameters measured simultaneously were analyzed for correlation and consis-tency with activated partial thromboplastin time(APTT),fibrinogen(Fib),and platelet count(Plt),and the parameters tested when ECMO was established and 24 hours after ECMO operation were compared.Results The correlation coefficient between R values and APTT of hmTEG(r=0.441,P<0.05)was lower than that of ordinary TEG(r=0.547,P<0.05).The parameters α-Angle and K value of ordinary TEG were not correlated with Fib(P>0.05),while as for hmTEG,the correla-tion was 0.359(P<0.05)and-0.343(P<0.05),respectively.The correlation between MA value of hmTEG and Plt was 0.456(P<0.05),which was much lower than its correlation with Fib(r=0.715,P<0.05).APTT and hmTEG had moderate agreement in judging the anticoagulant effect of UFH(P<0.05).Plt at 24 hours after ECMO was significantly lower than that at establishment of ECMO(P<0.05).Fib,APTT and hmTEG parameters were not significantly different between the two groups(P>0.05).Conclusion The parameters of hmTEG can better reflect the real level of coagulation factors in patients receiving ECMO.The results of hmTEG and APTT are complementary to assess whether heparin in ECMO patients is over-dosed,and hmTEG has unique advantages.Routine coagulation tests and TEG cannot replace each other,and the combina-tion of them can achieve better anticoagulation and coagulation management.

Objective To investigate the independent risk factors for acute severe cholangitis and related protective factors, and to construct a risk prediction scoring model for acute severe cholangitis. Methods A retrospective analysis was performed for the clinical data of 381 patients with acute cholangitis who were admitted to Department of Hepatobiliary Surgery, The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army, from January 2016 to July 2021, among whom there were 273 patients with non-severe cholangitis and 108 patients with severe cholangitis. Univariate and multivariate logistic regression analyses were used to screen out the independent risk factors for acute severe cholangitis and related protective factors, and then a logistic regression model was established. The receiver operating characteristic (ROC) curve was used to evaluate the discriminatory ability of the model, the calibration curve was used to evaluate the prediction accuracy of the model, and decision curve analysis (DCA) was used to evaluate the clinical value of the model. Moreover, the enhanced Bootstrap method was used to perform internal validation of the model and evaluate the performance of the model in internal validation. The model was visualized by the construction of Web calculator, nomogram, and scoring system. The two-independent-samples t test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. Results The univariate and multivariate logistic regression analyses showed that total bilirubin (TBil) (odds ratio [ OR ]=1.014, 95% confidence interval [ CI ]: 1.009-1.020, P < 0.001), percentage of neutrophils ( OR =1.128, 95% CI : 1.088-1.175, P < 0.001), and age ( OR =1.053, 95% CI : 1.027-1.082, P < 0.001) were independent risk factors, and albumin (Alb) ( OR =0.871, 95% CI : 0.817-0.924, P < 0.001) was a protective factor. The above independent risk factors and protective factor were included in the logistic regression analysis for model fitting, and the predictive model obtained had an area under the ROC curve (AUC) of 0.925 (95% CI : 0.897-0.952), with a specificity of 0.817 and a sensitivity of 0.935 at the optimal cut-off value of 0.245. The calibration curve showed that the predicted probability of the model was approximately equal to the actual probability, with a Brier value of 0.098, and the decision curve analysis showed that the model had a higher net income within the threshold probability interval of 0.1-0.9. Internal validation showed an AUC internal validation of 0.915 and a Brier value internal verification of 0.106. Conclusion TBil, percentage of neutrophils, and age are independent risk factors for acute severe cholangitis, while Alb is a protective factor. The established risk prediction scoring model has good discriminatory ability, calibration, and clinical value and can identify patients with acute severe cholangitis at an early stage, which provides a reference for subsequent diagnosis and treatment.