Inhibitory effect of survivin-shRNA on A431 cutaneous squamous cell carcinoma

Objective:To observe the effect of survivin-shRNA on A431 cutaneous squamous cell carcinoma(SCC)and explore the molecular biological mechanism of nuclear factorκB(NF-κB)in the inhibition of survivin-shRNA on cutaneous SCC.Methods:Survivin-shRNA adenovirus vector was constructed to screen out the best interference sequence.Nude mice were subcutaneously inoculated with the cultured A431 cell suspension to duplicate A431 transplanted tumor models.These mice were randomly divided into the blank control group,the rAd-EGFP negative control group,the rAd-survivin-shRNA transfection group and the Res positive control group,with 5 mice in each group.The corresponding reagents were injected into the tumors.All the mice were sacrificed on the 20th day.The tumor tissues were isolated,with tumor volume and tumor weight measured,the tumor growth curves were accordingly plotted and the tumor inhibition rate was consequently calculated.Hematoxylin-eosin(HE)staining was used to observe the cellular morphology of the tumors.TUNEL was applied to the detection of cell apoptosis.Western blot was applied to the detection of the expression of Survivin,inhibitor of NF-κB(IκB),P65,P53 and Caspase-3.Results:As to the transplanted tumors,tumor volume and tumor weight were decreased in nude mice in the rAd-survivinshRNA transfection group and the Res positive control group,in comparison with the blank control group and the rAd-EGFP negative control group,the differences were of statistical significance(p<.05);The results of TUNEL showed that the apoptosis rates were significantly increased in the rAd-survivin-shRNA transfection group and the Res positive control group,in comparison with the blank control group and the rAd-EGFP negative control group,the difference was statistically significant(p<.05);In the rAd-survivin-shRNA transfection group and the Res positive control group,the microscopic observation showed a small amount of sparsely-distributed tumor cells,degenerative liquefactive necrosis,cancer cell shrinkage and round-shape,and karyopycnosis;The expressions of Survivin and P65 proteins were decreased in the rAd-survivin-shRNA transfection group and the Res positive control group,in comparison with the blank control group and the rAd-EGFP negative control group,the differences were statistically significant(p<.05),while the expressions of IκB,P53 and Caspase-3 proteins were significantly increased,in comparison with the blank control group and the rAd-EGFP negative control group,the differences were statistically significant(p<.05).Conclusions:Survivin-shRNA can inhibit the growth of the transplanted tumors in cutaneous SCC nude mice,one of the mechanisms is to promote the apoptosis of tumor cells and inhibit the growth of SCC transplanted tumors by inhibiting the NF-κB signaling pathway and then activating the tumor suppressor gene P53.

Influence of thymopentin-5 on renal pathology and relevant indexes of serum in rats with acute kidney injury caused by sepsis

Objective:To discuss thymopentin-5 on renal pathology and relevant indexes of serum with sepsis-caused acute kidney injury(AKI)caused by cecal ligation and puncture(CLP).Methods:90 cases of healthy and male SD rats were randomly divided into 3 groups:control group(C group),sepsis group(S group),thymopentin group(T group).These groups were divided into five time point including 1 h,6 h,12 h,24 h and 48 h with 6 rats in each time point.The sepsis model was made by CLP.The blood and kidney tissue were collected in each time point.Changes of renal pathology were observed under light microscope and relevant indexes like serum creatinin(Cr),blood urea nitrogen(Bun),tumor necrosis factorα(TNF-α),interleukin-10(IL-10),CD4+/CD8+were tested and analyzed.Results:In T group,concentrations of Cr,Bun in 6 h after CLP started to rise,reached peak in 24 h and decreased in 48 h,which were all lower than S group(p<.05)and higher than C group(p<.05).Compared with C group,concentrations of TNF-αin 1 h significantly improved,reached peak in 12 h and decreased in 24 h,which were all lower than S group(p<.05);and concentrations of IL-10 in 1 h significantly decreased,reached peak in 12 h and rised in 24 h,which were all more than S group(p<.05).In T group,CD4+/CD8+ratio in 6 h after CLP started to decrease,reached the lowest in 24 h and rised in 48 h,which were all lower than S group(p<.05)and significantly lower than C group(p<.05).Conclusions:Thymopentin-5 plays the role of renal protection in AKI caused by sepsis.

Prediction of peritoneal free cancer cells in gastric cancer patients by golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging

Objectiveperitoneal free cancer cells can negatively impact disease progression and patient outcomes in gastric cancer. This study aimed to investigate the feasibility of using golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging (GRASP DCE-MRI) to predict the presence of peritoneal free cancer cells in gastric cancer patients.MethodsAll enrolled patients were consecutively divided into analysis and validation groups. Preoperative magnetic resonance imaging (MRI) scans and perfusion were performed in patients with gastric cancer undergoing surgery, and peritoneal lavage specimens were collected for examination. Based on the peritoneal lavage cytology (PLC) results, patients were divided into negative and positive lavage fluid groups. The data collected included clinical and MR information. A nomogram prediction model was constructed to predict the positive rate of peritoneal lavage fluid, and the validity of the model was verified based on data from the verification group.ResultsThere was no statistical difference between the proportion of PLC-positive cases predicted by GRASP DCE-MR and the actual PLC test. MR tumor stage, tumor thickness, and perfusion parameter Tofts-Ketty model volume transfer constant (Ktrans) were independent predictors of positive peritoneal lavage fluid. The nomogram model featured a concordance index (C-index) of 0.785 and 0.742 for the modeling and validation groups, respectively.ConclusionsGRASP DCE-MR could effectively predict peritoneal free cancer cells in gastric cancer patients. The nomogram model constructed using these predictors may help clinicians to better predict the risk of peritoneal free cancer cells being present in gastric cancer patients.