rAdinbitor was cloned from Gloydius blomhoffi brevicaudus in the laboratory. Previous researches had proved that rAdinbitor could inhibit proliferation of C6 glioma cells as well as promote their apoptosis. The molecular mechanism of rAdinbitor’s effects on C6 cells need to be further studied. rAdinbitor was expressed in E. coli BL21/pET23b-adinbitor and purified with Ni Sepharose 6 Fast Flow. The purified protein was confirmed by Western blotting. C6 cells were induced with fibronectin (FN). The effects of rAdinbitor with different concentrations on the expression of FAK, MEK1/2 and Caspase-3 as well as on activity of FAK and ERK1/2 in FN-induced C6 cells were studied by immunoblotting and immunoprecipitation. Results showed that rAdinbitor with different concentrations could obviously reduce the expression of FAK and MEK1/2, increase the expression of Caspase-3, as well as decrease ERK1/2 phosphorylation; besides 10 mg/L rAdinbitor, other concentrations’ rAdinbitor could inhibit FAK phosphorylation obviously. All those effects were dose-dependent. Results indicate that the effects of rAdinbitor on decreasing expression and activity of FAK and inhibiting Ras-MAPK signaling pathway play an important role in suppressing the proliferation of C6. Furthermore, the increase in Caspase-3 expression implies that the increase in apoptosis of C6 cells might be due to the suppression of rAdinbitor on the activity of ILK and PI-3K/Akt pathway.

Objective To explore the clinical treatment of patients with stage Ⅲ carcinoma of colon and rectum by tegafur chemotherapeutic drugs combined with standardized adjuvant therapy. Methods The data of 60 cases of carcinoma of colon and rectum treated in Wenzhou Central hospital from January 2013 to December 2015 were collected. The patients were divided into three groups, non standardized adjuvant treatment group of 18 cases, standardized adjuvant treatment group of 15 cases and tegafur drug combined with standardized treatment group of 27 cases. Recorded and compared the survival rate of the three groups within three years, the statistics of each group of disease-free survival (DFS) and overall survival rate (OS). According to the duration of treatment group, the use of oxaliplatin combined with 5-fluorouracil and leucovorin (FOLFOX) treatment, analyzed and recorded OS and DFS. Follow-up examinations were performed in patients who did not receive adjuvant therapy. Recorded and compared the OS and DFS of each group. Results Tegafur drug combined with standardized treatment group DFS and OS were significantly higher than the other two groups, the difference was statistically significant (P<0.05). In addition, when treated with FOLFOX regimen, the DFS and OS of group Ⅲ were significantly higher than those of groupⅠand groupⅡ, the difference was statistically significant (P<0.05). In the follow-up survey of three groups, the rates of recurrence and death in patients treated with tegafur drug combined with standardized treatment group were significantly lower than those in the non standardized adjuvant treatment group and the standardized adjuvant treatment group. Conclusion Tegafur combined with adjuvant therapy can significantly improve the standardization of the rate of carcinoma of colon and rectum and the rate of disease-free survival, it is suggested that doctors use chemotherapy combined with auxiliary standardized therapy, can significantly improve the survival of patients, and reduce the risk of disease recurrence rate.

Objective To study the relationship between expression of CD44v6 in gastric carcinoma and neoplasm metastasis and prognosis. Methods Expression of CD44v6 in 52 cases of gastric carcinoma was assayed by flow immunocytometry, and its relation with clinical pathology and prognosis was analyzed. Results In 52 cases of gastric carcinoma tissue, the positive rate of CD44v6 expression was 67.31%(35/52); but the positive rate of CD44v6 expression in normal gastric tissue was 25.00% (13/52). The positive rate of expression was significantly different (P

·AIM: To study the tear function changes in patients with pterygium before and after pterygium transposition.·METHODS: Twenty eyes of 20 patients were enrolled in this study.Schirmer I test and tear break-up time (BUT) were evaluated in patients before and after pterygium transposition.·RESULTS: There was no significant difference in the results of Schirmer I test before and after the surgery.The results of BUT were not significantly different before and up to 4 weeks after surgery. However, BUT prolonged significantly 6 weeks after surgery (P<0.05).·CONCLUSION: Pterygium transposition can improve the tear film function in patients with pterygium.

Objective To investigate the molecular mechanism of Loureirin A mediated anti-hepatic fibrosis by evaluting its effects on proliferation , secretion ofα-smooth muscle actin (α-SMA) and transforming growth factor-beta1 (TGF-β1), and expression of rat hepatic stellate cells in vitro . Methods Primary hepatic stellate cells were isolated and cultured from Sprague-Dawley rats. After activating and inducing primary hepatic stellate cells from qHSC to aHSC, the activated hepatic stellate cells model in vitro was established. Then we observed the morphological changes of static hepatic stellate cells and activated hepatic stellate cells with inverted phase contrast microscope. Cultured hepatic stellate cells were treated with different concentrations of loureirin A and the inhibitory rate of HSCs proliferation was measured by MTT assay. The expression of Frizzled-4 was measured by western blot analysis. The content ofα-SMA and TGF-β1 in the cultured HSCs'supernatant were measured by enzyme-linked immunosorbent assay (ELISA) . Results Loureirin A the proliferation of inhibited activated hepatic stellate cells in a time-dose-dependent manner compared with the control group,IC50=0.30 μg/μL. After loureirinA treatment of the HSCs, western blot analysis showed that Frizzled-4 expression level was obviously lower than control group. Loureirin A also inhibitedα-SMA and TGFβ1 (P<0.05) secretion in the cultured HSCs'supernatant in different degree by the assay of ELISA. Conclusions The molecular mechanism of Loureirin A and Wnt signaling pathway mediated anti-hepatic fibrosis and anti-angiogenesis may involve down-regulation the expression of Frizzled-4, inhibiting the synthesis and secretion ofα-SMA,TGF-β1and the proliferation of HSCs.

Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl₂) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl₂ was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L⁻¹ MnCl₂). They were treated with corresponding doses of MnCl₂ for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl₂ exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl₂ dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.

Objective: To evaluate the clinical efficacy of sinew-regulating bone-setting manipulations plus exercise therapy in treating chronic non-specific low back pain (CNLBP). Methods: A total of 65 CNLBP patients were divided into two groups by the random number table method. Thirty-three cases in the treatment group were intervened by sinew-regulating bone-setting manipulations plus exercise therapy; 32 cases in the control group were intervened by medium-frequency electrotherapy plus exercise therapy. Before and after treatment, visual analog scale (VAS), dynamic and static muscle endurance of low back, median frequency (MF) of surface electromyography (sEMG) and Oswestry disability index (ODI) were used to evaluate the low back function. The therapeutic efficacy was estimated after treatment. Results: The two groups each had 2 dropouts during the study. The total effective rate was 90.3％ in the treatment group versus 66.7％ in the control group, and the between-group difference was statistically significant (P<0.05). After treatment, the VAS score, dynamic and static muscle endurance of low back, MF of sEMG and ODI score all changed significantly in both groups (all P<0.05); all the items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Sinew-regulating bone-setting manipulations plus exercise therapy can effectively release pain in CNLBP patients, increase muscle endurance of the low back and improve the quality of life, and its therapeutic efficacy is more significant than that of medium-frequency electrotherapy plus exercise therapy.

Objective To evaluate the clinical significance of re operation for postoperative recurrent gastric carcinoma. Methods From 1986 to 2001, fifty one patients with postoperative local recurrence of gastric carcinoma were admitted into our hospital. The clinical data were analyzed retrospectively. Results Of 51 cases, there were 31 cases with recurrence within the stump stomach and 20 with local and metastatic recurrence. Twenty seven cases were treated by radical resection, 3 cases by palliative residual stomach resection, 15 cases by gastrojejunostomy or gastroenterostomy, 6 cases by simple exploration. Pathological examination of 30 cases revealed perianastomosis recurrence in 10 cases, stump stomach carcinoma in 20 cases. The 1,3,5 year survival rate of 27 cases after radical resection was 88%, 58%, 19% respectively. The survival time of palliative and comprehensive treatment group was 6 to 24 months and mean survival time was 16 months, while all patients undergoing simple exploration and abdominal cavity chemotherapy died after 2 to 7 months. Conclusion Most postoperative recurrent gastric carcinoma are within the residual stomach and hence could be treated by reoperative resection.

Objective: To observe the therapeutic effect of sinew-regulating bone-setting manipulations for knee osteoarthritis (KOA) model rabbits and its impacts on the chondrocyte apoptosis rate and the levels of interleukin (IL)-1β and nitric oxide (NO). Methods: According to the random number table method, 30 New Zealand white rabbits were divided into a normal group (n=9) and a modeling group (n=21). Rabbits in the modeling group were used to establish KOA models with the modified Hulth method. At the 8th week, three rabbits were sacrificed to verify the model and the remaining 18 rabbits were randomly divided into a model group (n=9) and an intervention group (n=9). Rabbits in the normal group and model group were bred routinely without any intervention. Rabbits in the intervention group were treated with the sinew-regulating bone-setting manipulations, 10 min/time, once every other day for a total of 20 times. The Lequesne MG knee function rating was used to evaluate the behavioral differences of the rabbits in each group. The Pelletier score was used to evaluate the general changes of the rabbits. The Mankin score was used to evaluate the pathology of knee cartilages. The enzyme-linked immunosorbent assay and nitrate reductase methods were used to determine the levels of IL-1β and NO in serum and synovial fluid of each group, respectively. In situ terminal deoxynucleotidyl transferase-mediated nick and labeling method was used to determine the apoptosis of chondrocytes in each group. Results: Compared with the normal group, the scores of Lequesne MG, Pelletier and Mankin, and the levels of IL-1β and NO in the model group were increased (P<0.05), which indirectly indicated the success of the model. Compared with the model group, the scores of Lequesne MG, Pelletier and Mankin, IL-1β and NO levels, and chondrocyte apoptosis rate of the intervention group were decreased, and the differences were statistically significant (P<0.05). Conclusion: The sinew-regulating bone-setting manipulations can reduce the levels of IL-1β, NO, and chondrocyte apoptosis rate, and delay the articular cartilage degeneration, therefore, having a good therapeutic effect on KOA.

Objective To investigate the neurotoxic effects of different concentrations of tetracaine and ropivacaine on the brachial plexus nerve in rats.Methods Forty-eight male Sprague-Dawley rats,weighing 410-430 g,were randomly divided into 8 groups (n =6 each):normal saline group (group NS),0.25％,0.50％ and 1.00％ tetracaine groups (groups T1-3 ),and 0.25％,0.50％,1.00％ and 2.00％ ropivacaine groups (groups R1-4 ).The rats received injection of normal saline 1.0 ml,0.25％,0.50％ and 1.00％ tetracaine 0.5 ml,0.25％,0.50％,and 1.00％ ropivacaine 1.0 ml and 2.00％ ropivacaine 0.5 ml in groups NS,T1-3 and R1-4 respectively through one side of the axillary sheath.The other side of the axillary sheath served as control side.Five days later,compound action potential and nerve conduction velocity (NCV) of the brachial plexus nerve were measured.Tne brachial plexus nerve was obtained as the specimen for microscopic examination with light and transmission electron microscope.Results Compared with the control side and group NS,the compound action potential and NCV of the brachial plexus nerve were significantly decreased in groups T2,3 and R3,4 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were gradually decreased with the increasing concentrations of tetracaine in groups T1 3 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were significantly decreased in group R4 as compared with groups R1-3 (P ＜ 0.05).The microscopic examination showed that the pathologic changes were more severe in groups T2,3 and R3,4 than those on the control side and than in group NS.Conclusion 0.50％ and 1.00％ tetracaine,and 1.00％ and 2.00％ ropivacaine can result in pathologic damage to the brachial plexus nerve in rats and the degree of damage is related to the concentration.