Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective: To study the relationship between the inhibitory effects of Tongxie Yaofang, a compound traditional Chinese herbal medicine, on the contraction of the colonic smooth muscle isolated from rats and calcium mobilization. Methods: By measuring the tension of the isolated colonic smooth muscle strips, the inhibitory effects of Tongxie Yaofang on the contraction induced by acetylcholine (ACh), KCl and exhausting Ca(2+) of internal calcium store were assessed respectively. Results: Tongxie Yaofang could concentration-dependently inhibit the contraction of isolated rat colonic smooth muscle strips induced by KCl and exhausting the Ca(2+) of internal calcium store. Tongxie Yaofang could also inhibit the tension of the second contractile phase induced by ACh (P<0.01, vs control), but had no influence on the first contractile phase. Conclusion: Tongxie Yaofang can inhibit the contraction of isolated rat colonic smooth muscle strips mainly by preventing the influx of extracellular Ca(2+), which may be associated with blocking voltage-dependent channel, store-operated channel and receptor-operated channel, but not by preventing the release of internal Ca(2+) from calcium store.

Objective To explore the clinical effects of variable angle locking plate ( VLP ) in the treatment of pediatric subtrochanteric femoral fractures. Methods Between January 2012 and November 2014, 9 pre-school children were treated at our department for subtrochanteric femoral fractures. They were 6 boys and 3 girls, with an average age of 4. 8 years ( from 4 to 6 years ) . By the Seinsheimer classification, 5 cases were of typeⅡB and 2 of typeⅡC and 2 of typeⅢA. The intervals between injury and surgery averaged 3 days ( from 2 to 5 days ) . All of them were treated with open reduction and VLP internal fixation. Results All the wounds healed well without any infection. All the stitches were removed within 12 days. They were followed up for 8 to 26 months ( average, 16 months ) . All the fractures united within 3 months after operation. Follow-ups revealed no plate or screw loosening, or refracture at the same site. According to the Beaty imaging criteria, the early outcomes were all satisfactory. At the final follow-ups, all the children gained normal gait after full-weight rehabilitation. The affected and normal hips are nearly identical in range of motion and muscle strength. All the children recovered their pre-injury status. By the Sanders scoring for function of the affected hip, 7 cases were rated as excellent and 2 as good. Conclusion VLP can be an effective option for treatment of subtrochanteric femoral fractures in preschool children patients.

Objective To investigate the effect of neuropeptide Y (NPY) on the osteoblastic differentiation of murine MC3T3-E1 cells and its mechanism related to the Wnt signaling pathway.Methods The murine MC3T3-E1 cells were divided into 4 groups according to the stimulators added:phosphate buffered saline (PBS) (control) and different concentrations of NPY (10-8 mol/L,10-10 mol/L and 10-12 mol/L).The cellular proliferation was detected with MTT assay after 1,3,5,7 and 9 days.The cells were identified with cell immunochemistry and Western Blot to find out the most effective concentration of NPY at different time points under osteoblastic condition.The cells were then divided into 4 groups:PBS,NPY,NPY + NPY receptor antagonist,and NPY + DKK1.Western blot was used to determine the expression of β3-catenin and p-GSK-3β in each group.Nuclear signaling activity of β3-catenin was observed using immunofluorescence staining.Results NPY significantly improved the proliferation of MC3T3-E1 cells at 7 and 9 days (P ＜0.05).NPY (10s mol/L and 10-10 mol/L) groups and NPY (10-10 mol/L and 10-12 mol/L) groups significantly improved the ALP activity at 4 and 14 days respectively (P ＜ 0.05).At 4 days,the expression of ALP protein was significantly decreased in the NPY + DKK1 group and the NPY + NPY receptor antagonist group compared with that in the NPY group (P ＜ 0.05).Although the expression levels of [β-catenin and p-GSK-3β protein were uninfluenced in either case,NPY significantly stimulated the nuclear signaling activity of β3-catenin.Conclusions NPY may significantly increase the expression of ALP protein in MC3T3-E1 ceils during osteoblastic differentiation.This effect might be mediated through the canonical Wnt signaling pathway.

Objective:To explore the prevalence of irritable bowel syndrome (IBS) based on a community population of Shanghai and the treatment of IBS by acupuncture therapy.Methods:A population of 1 685 subjects aged between 18-80 years were randomly selected by clustered sampling from the inhabitants in Changqiao Community of Shanghai to receive a questionnaire,for understanding the symptomatology and prevalence of IBS according to modified Manning and Rome II criteria,and the voluntary IBS patients were treated by acupuncture therapy.Results:The community- based prevalence of IBS was 11.45 % and 5.04% respectively according to modified Manning criteria and Rome II criteria.The ratio of male and female was 0.77:1 in IBS patients and the proportion was 10.62% and 12.19% respectively.But there were no significant difference in prevalence between different age groups (P＞0.05) .IBS is more common in subjects aged between 45-65 years (38.86%).Regarding the 62 IBS patients treated by electroacupuncture (EA),the total effective rate in Tianshu (ST 25) group (n=32) was 84.38%,and it was 56.67% in Daheng (SP 15) group (n=30),which showed a significant difference.(P＜0.05).Conclusion:IBS is a commonly encountered disorder in Changqiao Community of Shanghai and should be taken into consideration for human welfare,disease prevention and further epidemical investigation.Acupuncture treatment is recommended for treating IBS due to its satisfactory therapeutic effect.

OBJECTIVETo study the effects and mechanisms of sinensetin on proliferation and apoptosis of human AGS gastric cancer cells.

METHODMTT assay was used to detect the growth inhibition rates of human AGS gastric cancer cells treated with sinsesectin in different concentrations and times. The cell cycle distribution was measured by flow cytometry. The apoptosis was examined by Annexin-FITC/PI staining and DNA fragment analysis. The apoptosis morphology was observed by inverted fluorescence microscope after Hoechst 33342 staining. The protein expressions of p21 and p53 were detected by western blot.

RESULTMTT assay showed that sinensetin inhibited the growth of AGS gastric cancer cells in a dose- and time-dependent manner. Sinensetin blocked AGS cells in G2/ M and increased the apoptosis rates of AGS cells in a dose-dependent manner. DNA ladder was observed in cells treated with 60 micromol x L(-1) sinensetin for 48 h. The typical apoptotic morphological changes including cell nucleus shrinkage, chromatin condensation and apoptotic bodies were observed when treated with different dose of sinensetin. Western blot showed that sinensetin increased expressions of p53 and p21 in a dose-dependent manner.

CONCLUSIONSinensetin could inhibit human AGS gastric cancer cells proliferation and induce cell cycle block in G2/M phase and apoptosis. The up regulation of p53 and p21 protein might be one of the mechanisms.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; Dose-Response Relationship, Drug ; Flavonoids ; pharmacology ; Humans ; Stomach Neoplasms ; drug therapy ; pathology ; Tumor Suppressor Protein p53 ; analysis

Objective: To analyze the clinical features of perianal condyloma acuminatum in HIV infected men who have sex with men (MSM) and assess the effects of combined treatment of microwave and photodynamic therapy. Methods: Clinical and laboratory data of 260 male patients with perianal condyloma acuminatum were collected and microwave combined with photodynamic therapy was applied on them. Results: The majority of the patients with perianal condyloma acuminatum were young people (243/260). Warts were mainly located in the anus or from the outside to inside of the anus (245/260). Among them, the proportion of warts position of extraanal in HIV-infected group (8/220) was lower than that of non-HIV-infected group (7/40). HPV types 11 (70/118), 6 (44/118) and 16 (40/118) were the most common types of HPV infection. People with HIV infection were more likely to develop high-risk, mixed types HPV infection and multiple HPV infection than those without HIV infection. More than half of the patients (146/260) needed only 1-3 sessions of photodynamic treatments. There was no significant difference in the times of photodynamic treatments and the numbers of photosensitizers required between the HIV-infected and non-HIV-infected groups. The recurrence rate of HIV-infected group (13.6%, 30/220) was also similar to that of non-HIV-infected group (15.0%, 6/40), but far lower than those who underwent microwave therapy alone (45%, 18/40). Conclusions The combination therapy significantly reduced the recurrence rate of perianal condyloma acuminatum in HIV infected MSM.

OBJECTIVE: To determine the incidence and genotypes of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Dongguan region of Guangdong Province and assess the efficacy and feasibility of flow-through hybridization. METHODS: Peripheral blood samples were randomly selected and detected by modified G6PD/6PGD ratio method. Flow-through hybridization was used to detect 14 G6PD mutations among all samples. RESULTS: In total 1005 samples were collected, the detection rate for modified G6PD/6PGD ratio method and flow-through hybridization were 2.79% and 20.90%, respectively. The consistency of the two methods was poor(Kappa=0.187). When c.1311C>T mutation is excluded, the consistency of the two methods was good for males (Kappa=0.952) but still poor for females (Kappa=0.194). The most common mutations were c.1376G>T, c.1388G>A and c.95A>G. No G6PD deficiency was found among those only carrying the c.1311C>T mutation. CONCLUSION Flow-through hybridization can simultaneously detect 14 loci, covering over 90% of common mutations in Chinese population, and can be easily expanded. The routine method may miss many females carrying homozygous, compound heterozygous and heterozygous mutations, but the detection rate for male hemizygous mutation was much higher.
China ; DNA Mutational Analysis ; Female ; Genetic Testing ; Genotype ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; Humans ; Male ; Mutation

Objective To compare the diagnostic accuracy of magnifying chromoendoscopy (MCE) and endoscopic ultrasonography (EUS) for preoperative endoscopic assessment of the invasion depth of colorectal laterally spreading tumour(LST).Methods Data of 104 cases of colorectal LST were included.With the final pathological diagnosis as the golden standard,the accuracies of MCE and EUS for preoperative assessment of the invasion depth of colorectal LST were compared.Results The diagnostic accuracies of MCE and EUS for evaluating the invasion depth of LST were 89.4％ (93/104) and 73.1％ (76/104),respectively(P＜0.05).The lesion size and the endoscopist could affect the accuracy of the EUS evaluation (P=0.017,OR=3.561;P=0.035,OR =1.399).The accuracy of EUS seemed to show a downward trend for colorectal LST of larger diameters.Conclusion Both MCE and EUS are effective for evaluating the invasion depth of colorectal LST,but the accuracy of MCE may be higher than that of EUS.Large diameter of the lesion and the doctor's experience inadequacy may be the risk factors for the accuracy of EUS.

Sepsis is a critical illness with high morbidity and mortality. Anaerobic glycolysis plays an important role in the pathogenesis of sepsis. Pyruvate dehydrogenase complex (PDHC) serves as a key regulator during sepsis. With PDHC dephosphorylation and deacetylation, PDHC activity is upregulated, allowing pyruvate translocate to mitochondria in aerobic condition, preceding the production of acetyl-CoA to accelerate aerobic oxidation. Activation of PDHC improves the prognosis of sepsis through regulating the balance of lactate, release of inflammatory factors and energy metabolism. A variety of remedies can improve the prognosis of patients with sepsis by up-regulating the activity of PDHC, including dichloroacetate (DCA), vitamin B1, milrinone, tumor necrosis factor binding protein, and ciprofloxacin.This article reviews the role and the regulatory mechanism of PDHC and signal pathway in the sepsis metabolism, in order to innovate treatment for sepsis and multiple organ dysfunction.