AIM: To investigate the effect of LY980503(a benflumetol derivative)on multidrug resistance of tumor cell line using DNA microarray. METHODS: Total RNA was extracted from multidrug resistant MCF/DOX cell line. cDNA microarray containing 320 cDNAs was used to detect the gene expression profile. RESULTS: 9 down-regulated genes and 1 up-regulated gene were identified after multidrug resistant MCF/DOX cells were treated with LY980503. CONCLUSION: LY980503 can effectively reverse the resistance of MCF/DOX to DOX in vitro by adjusting the expression of multi-genes.

This study examined the effects of over-expression of leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) on the cell cycle and survival of human glioma cell line U87 and U251 and explored the possible mechanisms. The LRIG3 gene was transduced into U87 and U251 cells respectively by using lentivirus and the transduced cells were selected by puromycin. The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blotting. The apoptosis rate was detected by Annexin V-FITC/PI double labeling and the cell cycle was flow cytometrically analyzed. Compared with control cells, LRIG3 mRNA expression in U251 and U87 cells transduced with pLVX-DsRed-LRIG3-Monomer-N1 were increased by 77.6% and 129.7%, and LRIG3 protein expression was raised by 141.3% and 322.7%, respectively. Cell cycle analysis showed that LRIG3 over-expression increased the percentage of cells at G(0)/G(1) phase (P<0.01). Over-expressed LRIG3 could significantly promote the apoptosis of U87 and U251 cells (P<0.05). These findings suggest that the over-expression of LRIG3 could arrest the cell cycle in G(0)/G(1) phase, and promote apoptosis of U87 and U251 cells.

Background Curcumin can eliminate free radical arising in the system and superoxide anion and further inhibit lipid peroxidation.Researches proved that curcumin has resisting effect on oxidative damage of lens,but its effect on the formation of age-related cataract is under clear.Objective This study was to investigate the effect of curcumin on the cataract induced by sodium selenite.Methods Thirty healthy clean SD rats of 12-day-old were selected.All the rats were randomly divided into the normal control group,model group and curcumin group and ten rats for every group.The selenite cataract models were established in the rats of model group and curcumin group by the injection of sodium selenite subcutaneously.Meanwhile,0.005 ％ curcumin lavaged the rats in the curcumin group daily for two weeks.Lens of the rats were examined under the slit lamp at 4,7,10,14 days after injection in all groups,and the opacification of lens was graded based on the He' criteria.The lenses of rats were obtained at the end of experiment and then the level of superoxide dismutase (SOD),the activities of malondialdehyde (MDA) and glutathione peroxidase(GSH-Px) in rat lens were detected using biochemical assay method.The use of experimental animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The lenses were transparent throughout the experimental duration in the blank control group.Compared with model group,the development of the Ⅲ,Ⅳ,Ⅴ grade of cataract was slow down in the curcumin group with the significant difference( P＜0.05 ).The content of MDA,SOD and GSH-Px activity in lens appeared considerable differences among 3 groups ( MDA:F =215.42,P＜0.01 ; SOD:F =46.83,P＜ 0.01 ; GSH-Px:F=44.29,P＜0.01 ).The level of SOD was significantly reduced in the model group and the curcumin group compered with blank control group ( P ＜ 0.05 ) and that in the curcumin group was higher than the model group ( P＜0.05 ).The MDA level in the model group and the curcumin group was obviously rised in comparison with blank control group( P＜0.05 ),but that in the curcumin group lowed in comparison to model group( P＜0.05 ).GSH-Px activity in the curcumin group was significantly lower than that in blank control group and higher than that in the model group( P ＜ 0.05 ).Conclusions The curcumin can effectively delay rather than the cataract formation induced by sodium selenite in SD rat by improved the resistance of oxidation in the lens.

Objective To approach the efficiency and feasibility of preserving the fertility for patients with placental site trophoblastic tumor ( PSTT ).Methods Totally 2 086 cases of gestational trophoblastic neoplasm ( GTN) patients registered in Peking Union Medical College Hospital between 1998 and 2013.Fifty-seven of them were PSTT patients , 40 cases of which suffered hysterectomy , the rest 17 PSTT patients who preserved their fertility were analyzed retrospectively.The computerized database of clinical and pathological reports was reviewed in this cohort.Results The clinical manifestation of PSTT was not specific compared to other types of GTN.The average age of the 17 patients was 29.5 years old (range 22-39 years).The most common antecedent pregnancy was term birth (8 cases), the others were spontaneous abortion in 4 case, artificial abortion in 3 cases and molar pregnancy in 2 cases.The baseline serum β-hCG was slightly elevated and 12 patients (12/15) were less than 1 000 U/L.In this cohort, nine of the patients were in stage Ⅰ, while the other eight cases were in stage Ⅲ .The patients suffered conservative surgery, including dilation and curettage of uterus in 7 cases, open abdomen uterine lesion excision in 4 cases, laparoscopic uterine lesion excision in 3 cases, hysteroscopic uterine lesion excision in 1 case, and pulmonary lobectomy in 2 cases.Two of the patients didn′t received chemotherapy , while the other 15 cases suffered combination chemotherapy.Compared with 40 patients who suffered hysterectomy during the same interval , fertility preservation group did not result in poor outcomes or high risk of relapse rate.Six subsequent pregnancies happened after the therapy , two of them were during their second-trimester, while four patients had healthy babies by vaginal delivery in two and cesarean section in two .The scar of the uterus was fairly well during the cesarean sections.Conclusions Reservation of fertility therapy could be considered in highly-selected patients for young women who strongly desired to preserve their fertility and with localized lesion.Exactitude follow-up after therapy should be recommended.Contraception should also be recommended for at least one year after the chemotherapy.Vaginal delivery could be an option for the future pregnancies.

Objective:To analyze the characteristics of health financing at the provincial level according to the total health expenditure since China health system reform began in 2009 and provide evidence for improving health fi-nancing policy. Methods:20 provinces were chosen and vertical and horizontal Comparative approach was used to an-alyze the data. Results:Total health expenditure increased for all regions, of which the biggest rate was Anhui prov-ince, about 82. 97%, while the largest increasing for government health care expenditure was Ningxia province, a-bout 108 . 71%. In 2012 , the provinces with social health expenditure share of total above 40% were allocated in the east region, and the number of provinces with out-of pocket payment share of total above 40% reduces to 5. Conclu-sion:Total health expenditure grew in all regions, but there were differences in the degree that this spending matched the economic level;The financing structure was optimized, but the characteristic of regional financing was different. Some provinces were under huge pressure to reduce out-of pocket payments. Suggestions: Under the premise of im-proving the funding level, financing structure adjustment must be focused, and public funding should play a bigger role and out-of pocket payments should be reduced.

The effectiveness and safety of acupuncture for the treatment of supraventricular tachycardia were systematically reviewed. The randomized controlled trials (RCTs) regarding acupuncture for supraventricular tachycardia were searched in domestic and overseas databases, and the evaluation tool of bias risk in Cochrane Handbook 5.1.0 software was used to perform the evaluation of bias risk in literature, and RevMan 5.2 software was applied for statistics and Meta-analysis. Five RCTs involving 323 patients were included. The results showed that compared with the blank control group, the acupuncture reduced the heart rate by 18.8 times/min [95% CI (12.68, 24.92)]; the clinical effective rate in the acupuncture group was superior to that in the diltiazem group [OR= 3.11, 95% CI (1.50, 6.46)]; the difference of immediate effect between propafenone and acupuncture was not significant. No reports regarding adverse events was described in 5 RCTs. As was shown in the present evidence, acupuncture is safe and effective for the treatment of supraventricular tachycardia, but the level of evidence was low and the intensity of conclusion needed to be improved.
Acupuncture Points ; Acupuncture Therapy ; adverse effects ; Humans ; Randomized Controlled Trials as Topic ; Tachycardia, Supraventricular ; physiopathology ; therapy ; Treatment Outcome

OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged

Bacterial Infections ; diagnosis ; urine ; Colorimetry ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; diagnosis ; urine ; Urinalysis ; methods

Background The domestic and international researches discovered that many proteins and enzymes of the extracellular matrix (ECM) participate in the sclera remodeling by affecting the collagen typeⅠand fibronectin.Objective This study was to investigate the effect of matrixmetalloproteinase-2 (TIMP-2) on expression of collagen typeⅠand fibronectin of ECM in the posterior sclera by injecting liposomes containing tissue inhibitor of TIMP-2 gene into suprachoroidal space of the form-deprivation myopia in guinea pig.Methods Form-deprivation myopia was induced by translucent goggles in 36 clean guinea pig for 2 weeks.Then the animals were randomly assigned to TIMP-2 group,empty plasmid group,saline group and 12 for each group.Liposomes of 5μl containing TIMP-2 gene,empty plasmid and saline were suprachoroidally injected in the right eye respectively,and the left eyes without any treatment were used as self-control group.Other 12 matched guinea pigs only covered the right eyes through out the experimental duration as model control group.The guinea pigs were sacrificed and the posterior sclera tissue of the eyeballs were collected at 2,7 and 14 days after injection of drug.The expressions of collagen typeⅠmRNA and fibronectin mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).This study followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The expression level of collagen type Ⅰ mRNA in the posterior sclera of guinea pig was lower but that of fibronectin mRNA was higher in TIMP-2 group than self-control group,showing significant differences between them (P＜0.05).The expression level of collagen type Ⅰ mRNA in the posterior scleral tissue began to increase from the 2nd day after drug injection and was obviously elevated at the 7th day and then gradually decreased at the 14th day.However,the expression level of fibronectin mRNA in the posterior scleral tissue showed the opposite pattern.The expression levels of collagen typeⅠmRNA and fibronectin mRNA at the 7th after drug injection were significantly lower than that at the 2nd day or 14th day (P＜0.01).Conclusion Suprachoroidal injection of TIMP-2 in form-deprivation myopia could up-regulate the expression of collagen typeⅠmRNA and down-regulate the expression of fibronectin mRNA in the posterior scleral tissue.It may slow down the sclera remodeling of form-deprivation myopia in guinea pig in the early stage.

Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.