AIM:To investigate molecular regulatory mechanism of K-ras to girdin protein in COS7 cells and expression of K-ras and girdin in colorectal carcinoma tissues .METHODS:The lentiviral vector carrying K-ras gene was constructed and transfected in the COS 7 cells.The expression of K-ras, girdin proteins and other related proteins in COS 7 cells and colorectal carcinoma tissues was observed by Western blot .RESULTS:The COS7 cells with K-ras over-expres-sion showed an irregular cell morphology .The results of Western blot indicated that the downstream signal protein levels of p-ERK1/2, p-Stat3 and girdin were significantly increased in the COS 7 cells with K-ras over-expression.Transfection with the K-ras siRNA into the COS7 cells significantly reduced the protein levels of p-ERK1/2, p-Stat3 and girdin.In the color-ectal carcinoma tissues (7 cases), 5 cases had higher expression of K-ras and girdin compared with pericarcinous tissues . CONCLUSION:K-ras regulates girdin expression through the signal pathway of K-ras-ERK1/2-Stat3-girdin.

Objective To investigate the inhibition mechanism of sodium selenite on HCT116 cells.Methods In the present study,we explored the cytotoxicity induced by sodium selenite and the underlying mechanism by MTS assay,WesternBlot,and small RNA interference technique.Results It was found that the sodium selenite at 5uM concentration could indeed reduce the viability of colon cancer cell line HCT116 by a large margin through increasing the generation of reactive oxygen species (ROS),and that the increased levels of ROS could activate c-Jun Nh2-terninal kinase 1 (JNK1).Additionally,knockdown expression of JNK1 or p53 by using RNAi attenuated the cytotoxicity induced by sodium selenite,indicating that both of JNK1 and p53 are required in the process of cell death induced by sodium selenite.Conclusion The sodium selenite could induces cell death in HCT116 through oxidative stress by involvement of JNK1 and p53,both of which play a critical role in toxicity of sodium selenite.

BACKGROUND:Insulin-like growth factor-1(IGF-1) is an important regulator in osteoblasts proliferation,differentiation and matrix synthesis,which participates in the process of fracture healing and repairing. OBJECTIVE:To investigate the potential bone regeneration effect of human bone marrow mesenchymal stem cells(MSCs) transfected into IGF-1 retroviruses vector on bone defects repairing. DESIGN,TIME AND SETTING:A controlled observation experiment was performed between March 2004 and May 2005 at the Laboratory Animal Center of Soochow University. MATERIALS:MSCs were transfected into IGF-1 retroviruses vector in vitro and combined with demineralized bone matrix(DBM) . METHODS:Fifteen BalB/C nude mice were prepared for 8 mm calvarial defects models and divided into 3 groups by number table method,with 5 animals in each group. DBM transfected with IGF-1 and DBM transfected with vector were implanted into the defect in MSCs cells transduced with IGF-1 and vector groups,respectively. There was no other intervention in the blank control group. The implants were harvested and evaluated by histological examination at 4 weeks after model preparation. MAIN OUTCOME MEASURES:①The mRNA expression of IGF-1 was analyzed by RT-PCR and Western Blot. ②The complexes of cells and DBM were analyzed by scanning electron microscope(SEM) at days 3 and 6 after co-culture. ③Gross observation of the calvarial defects models. ④Osteogenic activity of the implants was evaluated by hematoxylin-eosin staining at weeks 4 after model preparation. RESULTS:①There were mRNA and protein expression of IGF-1 in MSCs cells transduced with IGF-1 vector. ②SEM showed that there were plenty of cells adhered to surface of DBM,and grew into inner part at the 3 days after co-culture with IGF-1,which secreted much collagen fibers at days 6. The count of adherent cells and collagen fiber was smaller in the vector group at each time points. ③No obvious inflammatory reaction could be seen in each group at 4 weeks after model preparation. Compared with vector control,abundant new bone and blood vessel formation occurred in the calvarial defects treated with DBM/IGF-1,and no new bone formation in the blank control groups. ④Results of hematoxylin-eosin staining showed that grafts in the MSCs cells transduced with IGF-1 group was combined closely with calvarial defects,new bone and vessel could be found. There were few bone-like materials formation in vector group,and no defects in control group were repaired. CONCLUSION:The composites of IGF-1 transfected MSCs and DBM has good effect in repairing calvarial defects.

Objective To observe the biological role and the underlying mechanisms of miR-132 in liver cancer on invasion and metastasis.Methods Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was used to examine the expression of miR-132 in four liver cancer cell lines (MHCC97H,HCCLYH,MHCC97L and SMMC-7721),a normal liver cell line HL-7702,and in liver tumor tissues with or without metastases.The biological effects of miR-132 transfection on human liver can-cer cells were assessed by wound assay,matrigel counting and in vivo experiments in nude mice.Western blotting was used to detect the expression of E-cadherin,α-cadherin,vimentin,fibronectin and ZEB2 in li-ver cancer cells.Immunohistochemistry was used to detect positive expression of ZEB2 in xenograft tumors.Results The expressions of miR-132 were downregulated in the four liver cancer cell lines when compared with the normal liver cell line (P ＜ 0.05),and in the liver cancer tissues with distant metastases when compared with the tissues without metastases (P ＜ 0.05).After transfection,ectopic expressions of miR-132 markedly inhibited cell migration and invasion in liver cancer cells.When compared with the control group,the expressions of E-cadherin and α-cadherin in the miR-132 transfection group were significantly increased,but the expressions of vimentin,fibronectin and ZEB2 were decreased.In addition,the numbers of metastatic lung lesions in nude mice in the miR-132 transfection group was markedly decreased when compared with the control group.The expressions of ZEB2 in the miR-132 transfection group was also significantly decreased when compared with the control group.Conclusions Transfection of miR-132 effectively inhibited invasion and metastasis of liver cancer cells in vitro and in vivo.miR-132 may become a new target for regulation of gene expression in liver cancer.

BACKGROUND:The important reason for failed back surgery syndrome is the postoperative epidural scar adhesions, therefore, exploring the methods of preventing postoperative lumbar epidural scar adhesions has always been a hot research in spine surgery field. OBJECTIVE:To investigate the effect of Shenshu point magnetic stimulation therapy on epidural scar adhesions in rat models of failed back surgery syndrome. METHODS:Sixty Sprague-Dawley rat models of failed back surgery syndrome were successfuly established using the method of laminectomy, and then divided into Shenshu magnetic therapy group and blank control group. Beginning from 1 week after modeling, rats in the Shenshu magnetic therapy group were subjected to bilateral Shenshu magnetic stimulation for 6 weeks, 5 days of treatment per week. Rats in the blank control group were not given any intervention. At 13 weeks after modeling, rats were harvested and the area ratio of epidural scars, range of adhesions and formation of colagen fibers were observed under light microscope. Fibroblasts were counted. Hydroxyproline content and transforming growth factor β1 expression in scar tissue were compared between these two groups.

Objective To study the effects of diazepam and sodium valproate infusion with CRRT in treatment of patients with tetramethylene disulfotetramine poisoning. Methods Diazepam and sodium valproate were continuously infused to patients by infusion pump. Meanwhile, CRRT was used to eliminate tetramethylene disulfotetramine. Results Convulsion was effectively controlled within one hour in 7 patients. After treatment with CRRT for 1 to 4 times, all patients recovered. Conclusion CRRT with diazepam and sodium valproate infusion were effective in patients with tetramethylene disulfotetramine poisoning.

Objective To explore the role of body fat distribution in the pathogenesis of obesity-related asthma.Methods Totally 125 patients with stable asthma were recruited and were divided into non-obese group (n =51),peripheral obesity group (n =34) and central obesity group (n =40) according to body mass index and waist circumference.The FEV1％,FVC,FEV1/FVC ratio,IL-6,and hs-CRP levels in peripheral blood,eosinophil and neutrophil percentage in induced sputum,as well as exhaled NO levels were determined,and asthma control test (ACT) scores were calculated.Both one-way analysis of variance and analysis of covariance were used for statistical analysis.Results The values of FVC in the central obesity group and the non-obese group were [3.98 (3.99) ±0.99] L and [4.51 (4.51) ±1.00] L,while the levels of IL-6 and hs-CRP in peripheral circulation and the percentage of neutrophils in induced sputum were [33.63 (33.28) ± 14.04] ng/L and [21.22 (21.33)±11.23] ng/L,[2.12 (2.15) ±0.73] mg/L and [0.92 (0.91) ±0.61] mg/L,52.58 (52.81) ± 14.14 and 45.41 (45.34) ± 12.84,respectively (all P ＜ 0.05).After adjusting for inhaled corticosteroids (ICS) doses,the ACT scores were also significantly higher in central obesity group (22.10 ± 1.68 vs.23.01 ± 1.62) (P ＜ 0.05).Only the hs-CRP level was found significantly higher in peripheral obesity group than in non-obese group [(1.54±0.68) mg/Lvs.(0.91 ±0.61) mg/L] (P＜0.05).Conclusion Central obesity may play the leading role in the pathogenesis of obesity-related asthma.

BACKGROUND:Cholecystokinin as an endogenous neuroprotective factor in the nervous system has garnered increasing attention. Findings from previous animal studies show that cholecystokinin can effectively promote the regeneration of the injured peripheral nerve. On this basis, further clinical trials wil be performed to observe whether local application of cholecystokinin at nerve anastomosis can promote peripheral nerve regeneration.
METHODS/DESIGN:As a prospective randomized controled trial, this study wil enrol 100 patients with complete rupture of the peroneal nerve, who wil be randomly divided into two groups: after nerve suture and partial gelatin sponge infiltration at nerve anastomosis, the patients wil be treated with 8 nmol/kg cholecystokinin (treatment group) or saline (control group). At 6, 12, 24 weeks after treatment, common peroneal nerve conduction velocity and electromyography and nerve fiber morphology wil be detected; the clinical efficacy at the last folow-up wil be assessed; and al adverse events during the folow-up wil be recorded to assess the therapeutic efficacy and safety.
DISCUSSION:In this study, cholecystokinin as an inducing agent for nerve growth factor synthesis wil be observed and studied, with a view to providing a new idea for seeking peripheral nerve therapy.
ETHICAL APPROVAL: The study protocol was approved by the Medical Ethics Committee of the Affiliated Hospital of Putian University (approval No. 2014116). Written informed consent wil be obtained from patients before treatment.

The emergence of atypical El Tor strains from V .cholerae in South Asia and Africa has been attributed to several outbreaks in recent decades ,however ,backgrounds of such strains in China remain exclusive .In this study ,PCR am-plification of both El Tor and classical alleles for ctxB ,tcpA ,rstR and hlyA genes was attempted in sixty-nine El Tor isolates from Fujian between 1962 to 2005 ,in addition ,some amplicons were sequence-analyzed .Thus ,the time point of atypical EVC strains in Fujian was determined ,genetic diversities of such strains were investigated .It was revealed that ctxB-Cl ,tcpA-Cl and hlyA genes were detected in O1 serogroup EVC isolates from Fujian since 1962 .Although rstR-Cl gene was solely detected in isolates between 1994 to 2000 .It was indicated by sequence analysis that atypical EVC strains from Fujian possessed a novel T→G mutation at residual 204 of the ctxB gene .Remarkably ,two novel ctxB genotypes (ctxB-10 and ctxB-11) were identified in one strain .The residual 115-C of ctxB in ctxB-11 showed characteristics of ctxB-Cl ,however ,its residual 203-T demonstra-ted characteristics of ctxB-El .This observation implied that it was common in O1 serogroup EVC strains from Fujian hybrid-ized with classical alleles since 1962 ,which would be the earliest time-point for the emergence of atypical El Tor strains hitherto in literature .Emergence of atypical El Tor strains harboring rstR-Cl in Fujian occurred since 1994 .Meanwhile ,novel mutation sites and ctxB genotypes were observed in Fujian isolates ,including diverse combination of ctxB genotypes in one strain and combination of biotype-specific sites in ctxB sequences .In summary ,molecular characterization of O 1 serogroup EVC strains from Fujian was unique and geography-associated .

Objective To investigate the risks of self-rated health in the ≥55-year elderly in Beijing and the occurrence of stroke.Methods The subjects (n=2 101;aged ≥55) from Beijing longitudinal study of aging (BLSA) were collected by Xuanwu Hospital,Capital Medical University from January 1992 to December 2016.One hundred and twenty-one subjects with stroke at baseline and 92 with incomplete information were excluded,and finally,1 888 elderly patients without cerebrovascular disease at baseline were included in the analysis.Based on the actual situation,the self-rated health was to identify an item that matched their current state from good,general to poor.The deadline for the survey was December 31,2012.The competitive risk model was used to assess the health self-rated status and the risk of stroke.Non-stroke deaths,including cancer and car accidents were treated as competitive events.Results Of the 1 888 subjects enrolled,946 (50.1%) self-rated health were good,616 (32.6%) were general,and 326 (17.3%) were poor;438 (23.2%) had stroke,751 (37.8%) had non-stroke death,and 699 (37.0%) were right censored data.Using the competing risk model and adjusting the age,sex,living area,marital status,education level,smoking,alcohol consumption,physical exercise,hypertension,diabetes mellitus,coronary heart disease,and body mass index,the occurrence of stroke in patients with poor self-rated health was 1.44 times (95%CI 1.11-1.87,P<0.01) as good as those who were good.Conclusion In the self-rated health of the elderly ≥55 years old in Beijing,the people with poor self-rated health increased the occurrence of stroke after considering the competitive risks.