Objective To study the effect of the recombinant vaccinia virus expressing human interleukin 2 on MKN45 gastric cancer cells in vitro. Methods Recombinant vaccinia virus expressing hIL 2 (VMJ601hIL 2) was constructed by homologous recombination using molecular virology, and VMJ601hIL 2 was detected by DNA hybridization technique and the recombinant gene product was analyzed by SDS PAGE. In addition, MKN45 gastric cancer cells was infected by VMJ601hIL 2 and the effect of VMJ601hIL 2 on the gastric cancer cells was evaluated in vitro. Results hIL 2 protein with biological activity can be secreted by MKN45 gastric cancer cells after heavily infected by VMJ601hIL 2. Conclusions It is one of the crucial steps that VMJ601hIL 2 has been constructed and identified since it forms essential prerequisite for further in vivo gene therapy of gastric cancer.

Objective To investigate the efficacy of injecting insulin in family diabetes mellitus in Xiaofeng town community in Huzhou area and the problems of injection technique.Methods From February 2015 to December 2016, 94 patients with diabetes were injected with insulin in Xiaofeng town community in Huzhou area.From February 2015 to 3016, the fasting blood glucose(FPG), 2 h postprandial blood glucose 2hPG) and glycosylated hemoglobin(HbA1c), and to investigate the problems in the process of injecting insulin by questionnaire.Results After 3 months of injection, FPG, 2hPG and HbA1c were decreased in all patients(P<0.05).71 cases(75.53) were regularly rotated in the injection of insulin and 82 cases were alcoholic(81.9%), the use of 5 mm needle 86 cases(91.49%)[our hospital only 5 mm and 8 mm Needle, jurisdiction of the surrounding hospital needle is 6 mm.(76.20%), 12 cases(12.77%) of insulin needle, and 2 to 3 days of insulin needle(72.60%) were injected in the subcutaneous 10s(71.28%) after injection, 16 cases(17.2%) were replaced, 10 cases(48.94%) were replaced on the 10th day, and 11 cases(11.70%) were replaced when the insulin was replaced.9 cases(9.57%) were replaced after the pain.Conclusion Diabetic patients have definite therapeutic effect on insulin injection in family, but there are some defects in injection technique and injection hygiene.Therefore, health care workers need to strengthen their knowledge of injection of insulin injections in families.

Objective To study the effects of extracellular-signal regulated kinase mitogenactivated protein kinase (ERK-MAPK) signaling pathway inhibition on histone phosphorylation and the related gene expression in human colorectal cancer cells.Methods Two human colorectal cancer cell lines (SW1116 and HCT116) were cultured and treated with gradient(0,20,40/μmol/L) doses of ERK-MAPK signaling pathway inhibitor U0126.Cell viability was determined by cell counting kit 8 (CCK-8) assay.Cell cycle distribution was assessed by flow cytometry.The expression levels of histone H3 kinases including ribosomal S6 serine-threonine kinase (RSK-2) and mitogen-and stressactivated protein kinase 1 and 2 (MSK1 and MSK2),and the levels of histone H3 (Ser10) phosphorylation and c-Fos protein were detected using Western blotting.Results Treatment of these two human colorectal cancer cell lines with ERK-MAPK inhibitor resulted in a time and dose-dependent inhibition of cell proliferation significantly. Proliferation rate of HCT116 was reduced to 47% at 72 hours after 40/μmol/L U0126 treatment. Cell cycle analysis showed that the percentage of phase G0/G1 cells significantly increased (P<0. 01) and the percentage of phase S cells decreased (P<0.01) after treatment with ERK-MAPK inhibitor. The expression of MSK1 and RSK2 reduced obviously in both of human colorectal cancer cell lines treated with U0126, which resulted in a 28% and 40% reduction of levels of MSK1 and RSK2 as compared with control HCT116 cells respectively,while no detectable change in the expression of MSK2 was found. Consistent with this, the expression level of histone H3 (ser10) phosphorylation was markedly down-regulated by ERK-MAPK inhibitor, and the related protein c-Fos expression decreased accordantly. Conclusions Decreased ERK-MAPK signaling pathway may reduce histone H3 (Ser10) phosphorylation via suppression of the activity of histone H3 kinase including MSK1 and RSK2, but not MSK2, consequently decrease the expression of c-Fos protein, which results in the inhibition of colorectal cancer cells proliferation.

Objective To determine the distribution of Helicobacter pylori (H.pylori) iceA, babA2 in patients in Shanghai and explore the association of H. pylori strain genotype with its clinical outcome after infection. Methods A total of 141 H. pylori strains was isolated from gastric biopsy samples of 43 patients with chronic gastritis, 47 patients with duodenal ulcer (DU), 30 patients with gastric ulcer(GU) and 21 patients with non cardia gastric carcinoma. The iceA, vacA, cagA, and babA2 genotypes were determined by polymerase chain reaction (PCR). Results iceA1, iceA2 and babA2 were detected in 74.5% (105/141) , 15.6% (22/141) and 63.8% (90/141) of the 141 H.pylori strains, respectively, while 2 of isolated H. pylori strain (1.4%) were positive for both iceA alleles and 16(11.3%) were negative for both iceA alleles. The prevalence of babA2 and the combined genotype of babA2 and cagA in H. pylori isolated from DU patients were significantly higher than that in GU patients (74.5% vs. 50.0% for babA2, P =0.028; 70.2% vs. 46.7% for babA2 and cagA, P =0.039). There was no significant difference in prevalence of babA2 among other disease groups. No association of different clinical diseases with iceA genotype was detected. Conclusions The most common genotype of H.pylori strains isolated from patients in Shanghai is iceA1 +/babA2 +. babA2 may play different role in the pathogenesis of duodenal ulcer and gastric ulcer. No association between iceA status and clinical outcome of H.pylori infection was confirmed in our study.

Objective To evaluate the safety and efficacy of 125I radioactive seeds implantation combined with postoperative chemotherapy as a treatment option for thoracic advanced esophageal squamous cell carcinoma(ESCC). Methods A prospective cohort study was carried out between 2000and 2005. According to preoperative CT staging criteria,298 patients in phase Ⅱ-Ⅲ of ESCC, who had were admitted to Oncology Center Surgery of Nanjing First Hospital Affiliated to Nanjing Medical University and Thoracic Surgery of YanCheng Oncology Hospital, were randomly divided into three groups: intraoperative 125I seeds implantation combined with postoperative chemotherapy (group A, 98cases), postoperative chemoradiotherapy (group B, 100 cases) and surgery alone (group C, 100cases). All patients received radical resection of esophageal cancer. According to pTNM staging criteria after operation, 233 patients in phase Ⅱb-Ⅲ of ESCC were finally enrolled in the study (78 in group A, 75 in group B, and 80 in group C). With 0. 5 m Ci of single seed, total activity of 5-11 mCi and matched peripheral dose in 60-70 Gy, 10-22 125I seeds were implanted into the target of patients in group A under direct vision in accordance with treatment planning system. The validation and quality assessment of radioactive seeds were demonstrated according to CT scan or X-ray imaging. The postoperative complications were observed. The local recurrence of the cancer was demonstrated using CT scan. The survival rate of patients was followed up for 1-,3-,5- and 10 years. Results The satisfied quality assessment of 125I seeds was observed. There was no displacement or loss of seed. The local recurrence in group A, B and C was 11. 5%, 13. 3% and 38. 8%, respectively, with statistical significance (P ＜ 0. 05). There was no significant difference among three groups with respect to complications and 1-year survival (P＞0. 05). However, the overall survival rate 3-, 5- and 10-years was 64.8%,37. 7% and 25. 1% in group A respectively; 63.3%, 36.9% and 24.9% in group Brespectively; 43. 6i%, 25.0%, and 12.6% in group C, respectively (all P＜0. 05). The 3-,5- and 10-year progression free survival rates were 63.5 %, 37.4 % and 15.1% in group A respectively; 62.5 %,36.6% and 14. 4% in group B respectively; 42.5%, 25.6% and 6.2% in group C respectively (all P＜0. 05). Conclusions It is a safe, effective and simple method for intraoperative 125I seeds implantation combined with postoperative chemotherapy in treatment of advanced ESCC, which may reduce the local recurrence and improve survival rates in patients with ESCC.

Hexanes ; adverse effects ; Humans ; Occupational Injuries ; chemically induced ; Pyrroles ; adverse effects ; Trauma, Nervous System ; chemically induced

Objective To analyze the effect of eukaryotic expression vector containing sense and antisense DNA methyltransferase (DNMT1) gene on the transcript level of tumor associated genes in human colon cancer cell line. Methods Recombinant plasmid, including sense DNMT1 (HMT) and antisense DNMT1 (THM) gene, were constructed and transfected into SW1116 cell by using the lipofectamine. Then G418 filtration was performed. The expression of DNMT1 protein was examined by Western blotting. The transcription level of hMLH1, hMSH2, c myc and p16 INK4A genes were detected by RT PCR. Results The sense and antisense eukaryotic expression vectors were successfully constructed and then the constructed recombinant plasmids were transfected into SW1116 cell. The protein levels of DNMT1 have been up regulated and down regulated in SW1116 cells transfected with HMT and THM plasmids, respectively. The mRNA level of hMLH1, hMSH2, c myc gene were down regulated in the sense DNMT1 transfected cell. The mRNA level of hMSH2 was up regulated in the antisense DNMT1 transfected cell. However, the transcription level of p16 INK4A gene could not be associated with DNMT1 in SW1116 cell.Conclusion DNMT1 can regulate the expression of the tumor associated genes in human colon cancer cell line SW1116.

Objective To investigate the effects of Helicobacter priori (H.prlori) strains from the patient with gastric cancer on the expression of DNA mismatch repair (MMR) genes in AGS cells in vitro. Methods AGS cells were co-cultured with ten strains of H.prlori from five patients with gastric cancer and five patients with gastritis respectively.The expressions of DNA MMR genes (hMSH2 and hMLH1) in mRNA and protein levels were determined by RT-PCR and Western blot.The enteropathogenic E.coli served as a bacterial control.Results E.coli and H.priori strains from gastritis have no effects on the expression of hMSH2 and hMLH1 in both protein and mRNA levels on the whole,while all the H.prlori strains from gastric cancer reduced expression levels of hMSH2 and hMLH1.Conclusions There are different effects between H. prlori strains from gastric cancer and those from gastritis on DNA MMR in AGS cell line,indicating that infec- tion of some H.prlori strains might lead to the inhibition of DNA MMR,and that in turns increases the risk of gastric carcinogenesis during chronic H.prlori infection.

Objective To explore a new method of functional reconstruction of hand digits and joints with free transfer of foot tissues so as to increase the success rate of the operation.Methods After micro-anatomic study of the plantar and dorsal metatarsal arteries,retrograde and free grafts of foot tissues pedicled with plantar metatarsal arteries were designed and applied in transplantation to treat 76 cases of hand digital or joint defects.The surgeries included 58 cases of transfer of the second toe,four cases of transfer of composite tissues of the second toe, eight cases of transfer of proximal interphalangeal joint,and six cases of nail flap transfer.Results The mi- cro-anatomic study found that the first plantar metatarsal artery was anatomically constant and the diameter of its branch to the second toe was larger than that of the first dorsal metatarsal artery.Flaps survived in 75 of the 76 patients(98.7%),with fine appearance and significantly improved function.One patient who had received free transfer of the second toe to reconstruct the thumb function had to undergo a second repair with infraclavicula skin tube because of refractory arteriospasm of anastomosed vessels.Conclusion Transfer with free retrograde grafts of foot tissues pedicled with plantar metatarsal artery to reconstruct hand functions can effectively improve the success rate of the operation,because it is free of the shortcomings of great anatomic variation of blood vessels and time-consuming and complex procedures in conventional transfer.

Objeetive To analyze the clinical efficacy of liver transplantation (OLT) for patients with hepatopulmonary syndrome (HPS).Methods From 2008 to 2013,420 adult patients underwent liver transplantation in our hospital.There were 91 patients with,and 329 patients without,HPS.The 5-year survival and mortality rates after OLT for the two groups were retrospectively analyzed.Results There were no significant differences between patients without and with HPS in age,primary disease,Child-Pugh score,MELD score,cold ischemia time and warm ischemia time.However,the differences on serum albumin [(29.6 ± 1.2) g/L vs.(26.4 ± 1.6) g/L] and blood oxygen pressure [(61.0 ±9.0) mmHg (1 mmHg =0.133 kPa) vs.(87.0 ± 6.0) mmHg] were significantly different (P ＜ 0.05).The 1-year cure rate was 65.9％ (60/91) in 91 patients with HPS after liver transplantation.The 1,3,5-year cumulative survival rates for patients without HPS were 97.3％,90.9％ and 80.3％,respectively,and the main causes of death were primary graft dysfunction,recurrent cardiovascular events and primary disease recurrence or tumors.The 1,3,5-year cumulative survival rates for patients with HPS were 65.9％,59.3％ and 56.0％,and the main causes of death were multiple-organ failure,pulmonary infection and cerebrovascular events.Kaplan-Meier survival curve analysis showed that the survival of patients with HPS was significantly lower than that of patients without HPS (P ＜ 0.05).Conclusions Liver transplantation is the most effective treatment for patients with HPS,but the short-term mortality rate is relatively high.We still need to learn more about HPS to improve the survival rate of patients with HPS after liver transplantation.