[Objective]To observe whether or not caffeine can improve the cyto-toxicity effect of methotrexate(MTX) on osteosarcoma cell line.[Method]Osteosarcoma cell(OS-732)were incubated with no drug,caffeine,MTX or MTX adding caffeine separately,and were named as control group,caffeine group,MTX group and mixing group respectively.The ratios of cells on G2M phase of each group were measured with flow cytometry after 48 h incubation and the cell inhibition ratios were measured with the MTT colorimetric analysis after 72 h incubation.[Result]The ratios(%)of cells on G2M phase were control group(23.210?0.416),caffeine group(23.120?0.440),MTX group(28.770?0.531)and mixing group(0.105?0.002)(P

[Objective]To study the difference in expression of RFC, GST-?, DHFRmRNA between human osteosarcoma U2-OS cell line and the MTX-resisitant variants U2-OS/R1-R3, and to investigate the significance in MTX resistance for human osteosarcoma. [Methods]Three resistant MTX human osteosarcoma cell lines were established by pulse exposure parental cell line(U2-OS) in gradually increased dose of MTX . The expression of RFC,GST-?,DHFRmRNA were assayed by real-time fluorescence quantitative polymerase chain reaction(FQ-PCR).[Results]Three MTX-resistant variants(U2-OS/R1-R3) were successfully established , the results of the FQ-PCR revealed that the MTX resistance was associated with the decreased expression of the RFC mRNA and increased expression of DHFR mRNA and GST-? mRNA.[Conclusion]The author investigated the MTX resistant mechanism of human osteosarcoma cell line at a gene level. The decreased expression of RFC mRNA and the increased expression of DHFR mRNA and GST-? mRNA participate in the MTX resistance in human osteosarcoma cell lines U-2 OS. This provides the evidence for exploring the MTX resistance mechanism in clinical osteosarcoma patients ,and helps to screen the patients who are insensitive to MTX chemotherapy.

[Objective]To investigate the expression of Survivin and its relationship with expression of PTEN and cyclinD1.[Method]Immonohistochemical method(SP method)was used to detect the expression of Survivin,PTEN and cyclinDl in 40 osteosarcoma tissues and 20 ostanchondroma tissues.[Result]The expression rate of Survivin was significantly higher in osteosarcoma(62.5%)than that in osteochondroma(10.0%)(P

Objective To know the current status of the household lavatories in the countryside of Guangxi Province, China and to present the scientific data for the government to make decision in the lavatory improvement in the countryside. Methods A stratified cluster sampling was used in the investigation and 2 414 peasant households in 241 villages were selected in 2006. Results The popularization rate of sanitary lavatory was 43.79% and 50.95% of the peasant households used the non-sanitary lavatory, 5.26% had no household lavatory. The main types of the sanitary lavatories used in the investigated area were firedamp tank-like and three-case-cesspool, accounted for 50.14% and 33.96% respectively. The rate of dejecta treatment for the non-sanitary lavatories was only 13.74%. As the money spent on the sanitary lavatory in each household was no less than 810 Yuan (RMB), maybe the popularization rate of the sanitary household lavatory could reach 50%. Conclusion Although the popularization rate of sanitary household lavatory in the countryside of Guangxi province significantly increased in recent years, it still does not fulfill the requirement for controlling the intestinal infectious diseases and parasitism in this area.

Objective To investigate the clinical value of the YSZ-1 uterine manipulator in total laparoscopic hysterectomy.Methods Total laparoscopic hysterectomy was employed in a total of 78 women by using the LigaSure,ultrasonic scalpel,and the YSZ-1 uterine manipulator.The anterior fornix was opened and the manipulator was placed on the lip of the cervix.Then the uterus was completely removed.Results The operation was completed smoothly in all the 78 patients.The operating time was 45~90 min(mean,63 min),and the intraoperative blood loss was 40~100 ml(mean,65 ml).No urinary tract injuries,vaginal bleeding,or other postoperative complications occurred.The length of postoperative hospital stay was 4~7 d(mean,5.3 d).Follow-up examinations in the 78 patients for 3~6 months(mean,4 months) showed that the vaginal wound normally healed up. Conclusions Use of the YSZ-1 uterine manipulator in total laparoscopic hysterectomy makes the procedure easier and safer.The YSZ-1 uterine manipulator is a useful laparoscopic instrument.

Objective To investigate the effect of cervical knife conization (CKC) or loop electrical excision procedure (LEEP)on the outcome of subsequent pregnancies and mode of deliveries. Methods A retrospective case-control study including 228 women after treatment with LEEP or CKC for cervical intraepithelial neoplasia (CIN) Ⅱ -Ⅲ who gave birth in the First Affiliated Hospital of Sun Yat-sen University and He-xian Memorial Hospital of Pangyu from January 2004 to January 2010 was performed.Patients (n = 228) without cervical surgical history were randomly extracted from the respective hospitals birth registries as controls and were matched by age, gestation,parity and income.The information including gestational age, premature rupture of membranes (PROM), type of deliveries and birth weight of the two groups were collected.Results The gestational age of women treated with conization was (268.3±26.2) d, longer than that of the women without surgery (279.4±25.3) d (t=4.60, P＜0.01). The incidence of preterm birth was 18.0％(41/228) and 4.4％ (10/228) (x2 = 21.22, P＜ 0. 05). The incidence of PROM was higher in conizationgroup (10.1％, 23/228) than that (1.3％, 3/228) in control group (x2=16.32, P＜0. 05). Risk for PROM was almost eight fold (OR=8. 42, 95％CI: 2.49-28.44) higher in conization group. Cesarean section rate was higher in conization group (69.3 ％ ) than in control group (39.0 ％ )(x2=42.06, P＜0. 01). The gestational age of women treated with LEEP was longer than those treated with CKC[(269.8±24.6) d vs (260.2± 26.5) d, t= 4. 01, P＜0.01]. The incidence of preterm birth was 13. 1％ (22/168) and 31.6％ (19/60) (x2 = 10. 34, P＜0. 05). The mean birth weight of women with LEEP was heavier than that with CKC[(3358.5 ±812.2) g vs (3295.9 ±832.6) g, t=3.08, P＜0. 01]. The incidence of PROM (7.1％, 12/168) of woman with CKC was higher than that (1.3％, 11/60) of women with LEEP (x2 =6.10, P＜0.05). Conclusions Conization might increase the incidence of preterm delivery and preterm PROM. LEEP showed less adverse effect onthe outcome of subsequentpregnanciesthan CKC,and waspreferredfor primigravida, and the risk of treatment should be informed in advance.

Objective To investigate the anti-arrhythmias mechanism of nardostachyos chinensis batal by intracardiac electrophysiologic study. Methods Intracardiac electrophysiologic study were performed in 40 supraventricular tachycardia(SVT)patients before and after oral nardostachys chinensis batal administraton,and the results were compared with 35 SVT patients or normal persons without nardostachys chineusis batal intake. Results The atrial effective,relative,functional refractory periods(AERP,ARRP,AFRP)and ventricular ERP(VERP)were significantly longer in nardostachyos chinensis batal administration group than control(AFRP(358 ± 76)ms vs.(256 ±69)ms;ARRP(365 ±74)ms vs.(287 ±55)ms;VERP(264 ± 36)ms vs.(213 ± 27)ms respectively);but the sinus and atrioventricular node function had no changes. Conclusions The anti-arrhythmia mechanism of nardostachyos chinensis batal were related to the prolonged cardiac refractory periods and the reduced cardiac excitability.

Objective To examine how ischemic time under common temperature affects acute rejection by using a rat vascularized skin transplantation model.Methods Vascularized groin flaps were transplanted from BN to Lewis rats with 1,2,3 and 4 h of ischemic time (Isc-1 h,2 h,3 h,4 h groups) under common temperature,and the allografts in each group were evaluated daily.Groin flaps were transplanted from Lewis to Lewis rats as control group.Biopsy samples taken from the each group on the postoperative day 2-8 were graded for signs of acute rejection.Biopsy samples taken from each group on the postoperative day 6 were stained for chemokine receptor CXCR3.Results When the ischemia time was 1,2,3 and 4 h,the survival time of the grafts was (9.0 ± 1.2),(8.6 ±1.1),(8.8 ± 1.3),and (7.0 ± 0.8) days respectively.The survival time in Isc-4 h group was significantly shorter than in other groups (P＜0.05).Histological evaluation showed acceleration of activated lymphocyte infiitration in the Isc-4 h group as compared with other g.roups.Furthermore,the proportion of CXCR3 positive cells in the Isc-4 h group was (50.1 ± 8.4) ％,significantly higher than that in the other groups on the day 6 after transplantation.Conclusion When ischemic time was over 3 h,the immune response is accelerated.The acceleration is associated with the higher expression of CXCR3 in the infiltrated cells.

OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors

Objective To investigate the current status, performance,challenges and demands of the health management service-related institutions in China so as to provide evidence for effective policy-making. Methods A cross-sectional study design was used in this report. The health management service-related institutions were identified based on web site searching and government business registration records. Web or mail-based questionnaire was used for data collection. Telephone interview was conducted to confirm the validity and reliability of the data. Results Of 5744 health management service-related institutions in China,585 were investigated for type of organization, personnel profile, business strategy and investment. Since 2000, the number of health management service-related institutions in China had kept increasing in an average rate of 52%. The services included physical check, health education, and disease intervention,etc. Conclusions In China,health management is still at its early stage although an increasing trend could be found and a large number of related- institutions have emerged. However, health management system in China still faces the challenge of poor professional services and insufficient healthcare providers.