BACKGROUND: Studies have demonstrated that trehalose possesses protective effects on cyropreserved sternum. But the mechanism of action remains poorly understood. OBJECTIVE: To investigate the effects of trehalose on bcl-2 and bax mRNA expression in cryopreserved sternum. METHODS: Four groups of freshly prepared solution were used: low-potassium dextran (LPD), LPD + dimethyl sulfoxide (DMSO), LPD + trehalose, LPD + DMSO + trehalose. Rat sternum was cut and then immediately cryopreserved in the tubes containing each group of solution. Fresh rat sternum tissue and 4 groups of samples cryopreserved for 120 days were taken and bcl-2 and bax mRNA expression in fresh and cryopreserved sternum was detected using reverse transcription-polymerase chain reaction.RESULTS AND CONCLUSION: bcl-2 mRNA expression in the LPD + trehalose group was significantly higher, but bax mRNA expression was significantly lower, than in the LPD, LPD + DMSO groups (both P < 0.01). LPD + DMSO + trehalose group showed highest bcl-2 mRNA expression and lowest bax mRNA expression, which were basically similar to fresh bone tissue (P > 0.05). These findings indicate that trehalose may protect cell activity in cryopreserved sternum by enhancing bcl-2 mRNA expression and inhibiting bax mRNA expression, and trehalose together with DMSO shows better protective effects.

Objective To construct a rapid genetic DNA extraction method, with nano magnetic beads, self-designed reagents system and extracting process. Method Part I: DNA extraction from old blood cotton swab sample with self-designed DNA extraction kit, then quantiifed by UV spectrophotometer. The method was further optimized on the preliminary results. Part II: All kinds of difficult DNA sample were tested with optimized kit, to detect the applicability of the kit. Result By improving the experimental condition, the extraction effects of different DNA sample is good, meanwhile, the extraction cost is relatively low.

To develop and validate a novel 6-dye STR(short tandem repeat) 25-plex DNA typing system for forensic DNA profiling and databases. In this study, a novel STR 25-plex DNA typing system that includes 24 autosomal STRs (D1S1656, D2S1338, D2S441, D3S1358, D5S818, D6S1043, D7S820, D8S1179, D10S1248, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11, D22S1045, CSF1PO, FGA, Penta D, Penta E, TH01, TPOX, vWA, D11S4463) and Amelogenin was developed. Validation studies demonstrated the sensitivity, accuracy, and reproducibility of our novel STR 25-plex DNA typing system. The sensitivity of the STR 25-plex DNA typing system was demonstrated by the ability to obtain complete profiles from as little as 0.125ng of human DNA. Specificity testing was demonstrated by the lack of cross-reactivity to a variety of commonly encountered animal species and microbial pool. For stability testing, full profiles were obtained with humic acid concentration ≤60ng/μL and hematin ≤600μM. For forensic evaluation, the selected 24 autosomal STRs followed the Hardy–Weinberg equilibrium. Since 24 autosomal STRs were independent from one another, PM (Probability matching) was 3.5434×10-28, TDP (Total Probability of Discrimination Power) was 0.999999999999999999999999969863, and CEP (Cumulative probability of exclusion) was 0.99999999375. The new STR 25-plex typing system is sensitive, reproducible, and stable, therefore it is highly applicable for use in national DNA database and can help to facilitate international data sharing.

OBJECTIVETo determine the feasibility and efficacy of endoscopic submucosal dissection(ESD) in treating early gastric cancer(EGC) and precancerous lesions in the remnant stomach of patients after gastrectomy.

METHODSClinical data of 36 patients with EGC and precancerous lesions in remnant stomach undergoing ESD in Endoscopy Center of Zhongshan Hospital from January 2008 to December 2013 were retrospectively analyzed. Operative, postoperative conditions and long-term follow-up of these patients were evaluated.

RESULTSBoth the success rate and the complete resection rate were 100%. The average maximum diameter of the tumor was 1.5(range 0.6-4.5) cm. During the ESD process, two bleeding cases were treated successfully by endoscopic hemostasis. The average operation time was 40(10-80) min. The delayed hemorrhage developed in 2 cases within 1-3 days after operation, and were also treated successfully by endoscopic hemostasis. There was no perforation or delayed perforation. No emergency surgery was required for the complication. Twelve cases were diagnosed as mild-moderate dysplasia, 7 cases as high grade intraepithelial neoplasia, 16 cases as hyperplastic polyps, and 1 case as signet ring cell carcinoma with T1 stage, who underwent operation for resecting gastric stump and lymph node dissection 7 days after ESD without subsequent follow-up. The curative resection rate was 92.7%(35/36). The median follow-up of the remaining 35 patients was 36(6-78) months without discomfort and recurrence under gastroscopy.

CONCLUSIONESD is safe and effective for EGC and precancerous lesions in the remnant stomach.

Adenocarcinoma ; Dissection ; Gastrectomy ; Gastric Mucosa ; Gastric Stump ; Gastroscopy ; Hemostasis, Endoscopic ; Humans ; Lymph Node Excision ; Neoplasm Recurrence, Local ; Operative Time ; Retrospective Studies ; Stomach Neoplasms

Objective To determine the feasibility and efficacy of endoscopic submucosal dissection(ESD) in treating early gastric cancer(EGC) and precancerous lesions in the remnant stomach of patients after gastrectomy. Methods Clinical data of 36 patients with EGC and precancerous lesions in remnant stomach undergoing ESD in Endoscopy Center of Zhongshan Hospital from January 2008 to December 2013 were retrospectively analyzed. Operative, postoperative conditions and long-term follow-up of these patients were evaluated. Results Both the success rate and the complete resection rate were 100%. The average maximum diameter of the tumor was 1.5 (range 0.6-4.5) cm. During the ESD process, two bleeding cases were treated successfully by endoscopic hemostasis. The average operation time was 40 (10-80) min. The delayed hemorrhage developed in 2 cases within 1-3 days after operation , and were also treated successfully by endoscopic hemostasis. There was no perforation or delayed perforation. No emergency surgery was required for the complication. Twelve cases were diagnosed as mild-moderate dysplasia, 7 cases as high grade intraepithelial neoplasia, 16 cases as hyperplastic polyps, and 1 case as signet ring cell carcinoma with T1 stage, who underwent operation for resecting gastric stump and lymph node dissection 7 days after ESD without subsequent follow-up. The curative resection rate was 92.7%(35/36). The median follow-up of the remaining 35 patients was 36 (6-78) months without discomfort and recurrence under gastroscopy. Conclusion ESD is safe and effective for EGC and precancerous lesions in the remnant stomach.

Objective To determine the feasibility and efficacy of endoscopic submucosal dissection(ESD) in treating early gastric cancer(EGC) and precancerous lesions in the remnant stomach of patients after gastrectomy. Methods Clinical data of 36 patients with EGC and precancerous lesions in remnant stomach undergoing ESD in Endoscopy Center of Zhongshan Hospital from January 2008 to December 2013 were retrospectively analyzed. Operative, postoperative conditions and long-term follow-up of these patients were evaluated. Results Both the success rate and the complete resection rate were 100%. The average maximum diameter of the tumor was 1.5 (range 0.6-4.5) cm. During the ESD process, two bleeding cases were treated successfully by endoscopic hemostasis. The average operation time was 40 (10-80) min. The delayed hemorrhage developed in 2 cases within 1-3 days after operation , and were also treated successfully by endoscopic hemostasis. There was no perforation or delayed perforation. No emergency surgery was required for the complication. Twelve cases were diagnosed as mild-moderate dysplasia, 7 cases as high grade intraepithelial neoplasia, 16 cases as hyperplastic polyps, and 1 case as signet ring cell carcinoma with T1 stage, who underwent operation for resecting gastric stump and lymph node dissection 7 days after ESD without subsequent follow-up. The curative resection rate was 92.7%(35/36). The median follow-up of the remaining 35 patients was 36 (6-78) months without discomfort and recurrence under gastroscopy. Conclusion ESD is safe and effective for EGC and precancerous lesions in the remnant stomach.

Objective To explore the role of NLRP3/Caspase-3 in myocardial apoptosis induced by ischemia/reperfusion(IR)injury and its effect on myocardiocyte autophagy in rats.Methods A total of 60 SPF-grade male rats were randomly divided into sham operation,model,and nimodip-ine treatment groups,with 20 rats in each group.Rat model of myocardial IR injury was estab-lished in the rats of the two latter groups.Cardiac function was assessed,and the levels of myocar-dial enzymes and cytokines were measured.Additionally,myocardial pathological changes were de-tected using HE staining.Furthermore,flow cytometry was utilized to determine the apoptotic rate of myocardiocytes,and the autophagosomes were counted under transmission electron micro-scope.Moreover,the expression of NLRP3 and Caspase-3 was measured using RT-PCR and West-ern blotting.Results Significant differences were observed in left ventricular end diastolic pres-sure,left ventricular systolic pressure,maximal rate of rise and fall in left ventricular pressure,ap-optotic rate of myocardial cells,and levels of TNF-α,IL-6,CK,AST and LDH in the three groups(P＜0.01).Notably,both the model group and nimodipine treatment group exhibited significantly higher autophagosome than the sham operation group(10.55±1.87 and 6.32±1.43 vs 3.45±0.67 units,P＜0.01),and the nimodipine group displayed a significantly lower autophagosome count than the model group(P＜0.01).The mRNA and protein levels of NLRP3 and Caspase-3 were notably higher in the model group and nimodipine group than the sham operation group(P＜0.01),and in the model group than the nimodipine group(P＜0.01).Conclusion Myocardial IR injury in rats can increase myocardiocyte apoptosis,reduce cardiac function,induce inflammatory response,and enhance autophagosome formation,which is related to the abnormal high expression of NLRP3/Caspase-3.

Objective To explore the impact of knocking down Sec31A on the malignant phenotype of head and neck squamous cell carcinoma(HNSCC)and its possible mechanisms.Methods Transcriptome sequencing data of HNSCC tissues and adjacent tissues were obtained from the TCGA database,and the expression levels of Sec31A were compared.Immunohistochemical staining was used to analyze the expression of Sec31A in HNSCC tissues.Kaplan-Meier survival analysis was used to compare the relationship between Sec31A and the prognosis of HNSCC patients.Small interfering plasmids si-Sec31A and si-NC were transfected into HNSCC cell lines HN6 and HN4,and the impact of knocking down Sec31A on the biological behavior of HNSCC cells was detected through CCK-8 exper-iments,plate cloning experiments,scratch healing experiments,and Transwell experiments.Changes in the expression levels of PI3K/AKT/mTOR pathway related proteins in cells were detected after knocking down Sec31A with HN6 and HN4 through Western Blot(WB)experiments.Stable transfected cell lines of HN6 siSec31A and HN6 siNC were constructed and inoculated subcutaneously in nude mice to further verify the tumorigenic effect of Sec31A in vivo.Results TCGA data showed that Sec31A was higher in HNSCC tissues than in adjacent normal tissues(P＜0.01),and high expression of Sec31A was significantly correlated with poor prognosis in pa-tients(P＜0.05).Immunohistochemical staining showed that Sec31A was expressed stronger in HNSCC tissues than in normal tissues.In HN6 and HN4 cells,knocking down Sec31A resulted in significantly weaker proliferation,migration,and invasion abilities compared to the control group.Through WB experiments,it was found that transfection of si-Sec31A with HN6 and HN4 significantly reduced the expression levels of p-PI3K,p-AKT,and p-mTOR proteins.After knocking down Sec31A with HN6,the transplanted tumor volume in nude mice was significantly smaller than that in the control group.Conclusion Knocking down Sec31A can inhibit the proliferation,migration and invasion of HNSCC cells,possibly through the PI3K/AKT/mTOR pathway.