China ; Health Surveys ; Humans ; Physical Fitness ; Students

Objective To study the expression and clinical significance of Notch3 and Notch intracellular domain (NICD) in ovarian carcinoma and the effects of N-[N-(3 ,5-difluorophenyl) acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis in OVCAR3, A2780 ovarian carcinoma cell lines. Methods Western blot was used to detect the expression of NICD in the tissues from 58 ovarian carcinomas patients and 21 normal ovarie, who were admitted in Peking University First Hospital from July 2006 to June 2009. Immunohistochemistry was also used to detect the expression of Notch3 in these tissues. The relationship with clinical features of ovarian carcinoma was also analyzed. Proliferation of OVCAR3 and A2780 ovarian cancer cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in OVCAR3 and A2780 cells incubated with DAPT was detected by western blot. Results (1)The expression level of NICD in ovarian carcinomas was significantly higher than that in normal ovarian tissues (1.64 ±0. 19 vs. 0.98 ±0.20;P <0.05). The NICD expression was higher in ovarian cancers with low grade or advanced stage than those in high-middle grade or early stage,respectively (1.90 ± 0. 22 vs. 1.25 ± 0. 21,1.80 ± 0. 21 vs. 1.21 ± 0. 15; all P < 0. 05). The Notch3 protein was stained positively in cytoplasm, nuclear and cell membrane. The expression of Notch3 was higher in ovarian carcinomas than that in normal ovaries [78% (45/58) vs. 24% (5/21); P < 0. 01]. While,there were no stasistical difference in different pathological types, stages, differentiation of ovarian carcinoma. There was no difference between the patients with adjuvant chemotherapy or not. (2)After OVCAR3 and A2780 cells incubated with DAPT 24, 48, 72 hours, NICD expression was significantly lower than that in control group (P < 0. 05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells were depended on the concentrations and times. Conclusions Notch3 and NICD may play a key role in the occurrence and progress of ovarian carcinoma. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells may be due to decreased the formation of NICD.

Objective To study the effect of allicin on the growth and radiosensitivity of human pancreatic carcinoma BXPC3 cells.Methods BXPC3 cells were exposed to X-rays in the presence or absence of allicin.Cell proliferation was measured by MTT assay.Cell cycle distribution and apoptosis were detected by flow cytometry assay.Cell radiosensitivity and the influence of allicin on it was evaluated by colony formation assay.The expressions of Bax and Bcl-2 proteins were assayed by RT-PCR and Western blot.Results IC50 values of allicin on cell growth were 76.24,58.34 and 43.58 μmol/L under 12,24 and 48 h treatment,respectively.Treatment of cells with allicin obviously inhibited cell growth after irradiation and hence increased radiosensitivity (t =2.74,P ＜ 0.05).This treament also enhanced radiation-induced cell cycle arrest at G2/M phase (t =11.41,P ＜0.05),apoptosis induction (t =12.36,P ＜ 0.05),and Bax expression (t =4.83,P ＜ 0.05),but it decreased Bcl-2 expression (t =3.69,P ＜ 0.05).Conclusions Allicin could inhibit cell growth,induce cell cycle arrest and apoptosis via Bax/Bcl-2 pathway and hence increases radiosensitivity of BXPC3 cells.

Objective To investigate the diagnostic value of B-mode ultrasonography in detecting middle hepatic vein(MHV)in chronic liver disease patients.Methods 80 chronic liver disease patients were divided into 2 groups(chronic hepatitis and liver cirrhosis).Liver biopsies and the inner diameter(ID)of MHV was detected with B-mode ultrasonography.The ID of MHV was compared in the chronic hepatitis and liver cirrhosis groups,different liver fibrosis stages and compensation/non-compensation liver cirrhosis.The ability of ID of MHV in auxiliary diagnosis liver cirrhosis was analyzed with the receiver operating characteristic curve(ROC).Results The size of ID of MHV in liver cirrhosis(3.82±1.84)mm was smaller than that of chronic hepatitis(6.15±1.67)mm(P＜0.01).The size of ID of MHV in non-compensation liver cirrhosis(2.98±1.15)mm was smaller than that of compensation liver cirrhosis(4.42±2 20)mm(P＜0.05).There was midrange negative correlation with liver fibrosis stages and the ID of MHV(rs=-0.465,P＜0.01).The cutoff point of ID of MHV diagnosis liver cirrhosis was 4.7mm.The area under ROC(AUC)achieved 0.813(P＜0.01).The sensitivity(Se),specificity,(Sp),positive predictive value (PPV),negative predictive value(NPV)and Youden index were 67.5%,90.0%,88.0%,73.5% and 57.5%,respectively.Conclusion There is well clinical value with B-mode ultrasonography detecting ID of MHV for the auxiliary diagnosis of chronic liver disease.

Objective To investigate the effects of postoperative adjuvant chemotherapy (CT) and chemoradiotherapy (CRT) or radiotherapy(RT) for Ⅰ b-Ⅱ a cervical cancer with risk factors.Methods From March 1995 to June 2010,there were 137 patients underwent radical hysterectomy and systematic pelvic lymphadenectomy for stage Ⅰ b-Ⅱ a cervical cancer admitted at Peking University First Hospital.These patients had risk factors,intermediate risk factors including bulky tumor (＞4 cm),lymph vascular space invasion,deep stromal invasion; high risk factors including positive surgical margin,parametrial invasion,lymph node involvement.Of the all patients,79 cases of them were treated with CT,58 of them were treated with RT or CRT.The 5-year survival and prognosis factors were analyzed retrospectively,the prognosis was compared between two adjuvant therapy groups.Results The univariate analysis shown that types of pathology,different grade of risk factors,stroma invasion and lymph node involvement were prognostic factors of 5-year overall survival Patients with squamous cell carcinoma,intermediate risk factors,no parametrial invasion,and no lymph node involvement had better prognosis (P ＜ 0.05).Whether patients with high-risk factors or intermediate-risk factors,the 5-year overall survival and 3-year disease-free survival had no difference between CT and RCT or RT groups respectively.Cox regression multivariate analysis of survival indicated that clinical stages,types of histology,different grade of risk factors were independent prognostic indicator.Patients with early stage,squamous cell carcinoma,intermediate risk factors had better prognosis.Univariate and multivariate analysis indicated that different postoperative adjuvant therapies had no effects on the prognosis.The 5-year overall survival was 88.6％ in patients treated with CT,and 89.7％ in patients treated with RT or CRT (P =0.455).Conclusion There are equivalent therapeutic results between CT and RT or CRT for patients with risk factors after radical surgery,CT may be as one choice of postoperative adjuvant therapy for stage Ⅰ b-Ⅱ a cervical carcinoma with risk factors.

By summarizing the recent literatures on brain mechanisms with acupuncture intervention based on blood oxygenation level-dependent (BOLD)-functional magnetic resonance imaging (fMRI), the BOLD-fMRI examination and analysis methods, the points to be acupunctured, the corresponding meridian activation regions, the specific intensity range, functions and indications of the acupoints, the manifestation of 'bi-directional regulation' characteristics, fMRI performance of chronergy, laterality and needling qi of acupuncture were reviewed to provide the ideas for future research in this area.

OBJECTIVETo establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field.

METHODSThree 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining.

RESULTSCompared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01).

CONCLUSION4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Chloral Hydrate ; pharmacology ; Cilia ; drug effects ; enzymology ; physiology ; Male ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley

Essential hypertension (EH) is affected by both genetic and environmental factors.The polymorphism of connexin (Cx) genes is found associated with the development of hypertension.However,the association of the polymorphism of Cxs with EH has not been investigated.This study aimed to investigate the association of the polymorphism of connexin (Cx) genes Cx37,Cx40,and Cx43 with EH in Kazak and Han Chinese in Xinjiang,China.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were used to analyze the polymorphism of Cx genes in Kazak and Han EH patients as well as their normotensive controls.The results showed that there were no significant differences in the frequencies of different three genotypes (A/A,A/G,and G/G) and A and G alleles of Cx40 rs35594137 and rs11552588 between EH patients and normotensive controis.However,in Kazak EH patients,the frequencies of three genotypes (A/A,A/G;and G/G) of Cx37 rs1630310 were 24.8％,47.2％ and 28.0％,respectively,which were significantly different from those in Han EH patients.In Han EH patients,the frequencies of the three genotypes (C/C,C/G and G/G) of Cx43 rs1925223 were 6.4％,35.6％ and 58.0％,respectively.Frequencies of the other four genotypes had no statistical differences among Kazak and Hart EH patients and their normotensive controls.These results suggest polymorphisms of Cx37 rs 1630310 and Cx43 rs 1925223 genes may be associated with the pathogenesis of EH.Carrying Cx37 rs1630310-A or Cx43 rs1925223-G genotypes may protect against the development of EH.

OBJECTIVETo investigate osthole effect on femoral tissue resorption activity of rat in vitro.

METHODSSix SD rats weighted (80 ± 5) g were used to isolate and culture femoral tissue (diaphyses and metaphysis) in vitro. The cultured tissue were devided into control group, estradiol group and osthole group. The femoral tissue was treated with final concentration of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol culture in vitro at 48 hours after cultured. Tartrate-resistant acid phosphatase (StrACP) activity, glucose and Lactic acid content, StrACP, MCSF (Macrophage colony stimulating factor) and CTSK (Cathepsin K) mRNA was detected by Real-Time RT-PCR were detected.

RESULTSConcetration of Alkaline phosphatase activity were 2226 and 2498 in 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol respectively. As compared with control group, the activity of StrACP of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol were inhibited at 6, 9, 12 days (P < 0.05); under treatment of in l x 10(-5) mol/L osthole, the content of Lactic acid were increased and the content of glucose were decreased at 3, 6, 9 days (P < 0.05); StrACP, MCSF and CTSK mRNA expression level were inhibited at 6, 9 days (P < 0.05).

CONCLUSIONOsthole can inhibit bone resorption and raise the level of nutrition metabolism of femurs tissue.

Acid Phosphatase ; metabolism ; Animals ; Bone Resorption ; prevention & control ; Coumarins ; pharmacology ; Estradiol ; pharmacology ; Femur ; drug effects ; Glucose ; analysis ; Lactic Acid ; analysis ; Male ; Rats ; Rats, Sprague-Dawley

A strain KX6,producing thermotolerant endoglucanase,was isolated from compost. The morpholo-gical identification and 16S rRNA sequence analysis showed it belongs to Streptomyces xylophagus. The production and characterization of endoglucanase from Streptomyces xylophagus KX6 was studied. Maximum endoglucanase yield of 0.538 IU/ml was achieved with medium pH8.0,containing CMC2Na 1.0% as carbon resource,soybean meal 1% as nitrogen resource,2% inoculating volume,30% 250 ml triangle flask bulk for medium volume at 40℃ 200r/min shaker for 48h. The endoglucanase exhibited optimum catalytic activity at pH7.0 and 50℃. The enzyme was stable at 50℃,and able to retain 60% of the full activity,when it was incubated at 60℃ for 1h.The enzyme was stable at pH6.0~7.0. All these findings suggest that the enzyme is a thermotolerant neutral endoglucanase.