BACKGROUND: Under the pressure of mechanical action, the culture membrane will be stretched and cause the deformation of cells which adhere to the surface of culture membrane. According to this, growth regularity in different external force environments will be observed; however, how to determine the load distributions on the whole culture membrane still remains unclear. OBJECTIVE: Photomechanics method was applied for determining the out-plane displacement so as to obtain the strain distributions of the cell culture membrane. DESIGN, TIME AND SETTING: Contrast study was performed at the Photomechanics Laboratory, School of Mechanical and Automation Engineering, Shanghai Institute of Technology from March to August 2008. MATERIALS: Culture membrane which specially used for biomedicine was made by Dow Coming Corporation, USA and the type was Q7-4750. The dimension and mechanics properties were as follows: diameter = 100 ram, thickness = 150-160 μm, modulus of elasticity E = 2.14 MPa, and Poison's ratio = 0.48. lines per millimeter; phase shift set was controlled by manual and 5 marks equal 0.1 mm; CCD camera was used for capturing moire patterns. Finally, phase was transformed into displacement so as to obtain three-dimensional appearance and manner. MAIN OUTCOME MEASURES: Out-plane displacement information, and deflection of culture membrane including displacement and strain. RESULTS: The three-dimensional profiles of culture membrane after deformation were constructed by image processing method and the out-plane heights could be obtained. Corresponding to the total strains of 1%, 10%, 20%, and 25% for the culture membrane, the displacement of highest point was 2.28 mm, 8.32 mm, 12.12 ram, and 13.52 mm, respectively. The error was 3.5% through comparing the measured heights of culture membrane with the heights which was known for the culture membrane. It indicated that this experiment method was highly sensitivity. According to the out-plane displacements, the strains distributions along symmetrical axis were calculated and the distributions of strain were shown by parabolic curve. displacement distributions of culture membrane is obtained and shown displacement contour. Namely, the out-plane displacement of center for the membrane has the maximum heights, and the strains distributions are shown parabolic curve.

Objective:To identify the relationship between expression of protein kinase R(PKR), phosphating PKR, EIF2α(p-PKR, p-EIF2α) in PKR→EIF2α signal transduction passage and the grades of cervical lesions, the role in generation and progression of cervical tumor and their effects to prognosis of cervical cancer patients. Methods:The expressions of PKR, p-PKR and p-EIF2α in human cervical cancer tissue of 63 cases, cervical intraepithelial neoplasia(CINⅠ-Ⅲ) of 114 cases and normal cervical epithelium of 15 cases were detected by immunohistochemical technique. Results:With the increase in grades of cervical lesions, the positive-expression rate of PKR increased and significantly correlated with the grades of cervical lesions(P < 0.05). With the increase in grades of cervical lesions, the positive-expression rates of p-PKR and p-EIF2α increased firstly, and then decreased. In cervical cancer group, the positive-expression rate of PKR was much higher than that of p-PKR(P < 0.01). The development and progression was quicker in later clinical stages of cervical cancer than that of earlier clinical stages of cervical cancer (P < 0.01). The development and progression of cervical cancer was quicker in patients with negative-expression of p-PKR and p-EIF2α than that in patients with positive-expression of p-PKR and p-EIF2α(P < 0.05). Conclusion:The positive-expression rate of PKR was correlated with the grades of cervical lesions. There are some factors which can impede PKR and EIF2α to be phosphorylated or make p-PKR and p-EIF2α dephosphorylate in high level cervical lesions, which promotes the development and progression of cervical lesions, worsens the prognosis of cervical cancer.

Objective To explore the relationship of signal transduction among human papillomavirus 18 E6 oncoprotein (HPV18E6), signal transducers and activators of transcription 1 (STAT1), protein kinase R( PKR )/α subunit of eukaryotic initiation factor 2 ( eIF2α ), nuclear factor-kappa Bp65 ( NF-κBp65 ), mitogen-activated protein kinase( MAPK)/c-Jun N-terminal kinase(JNK) ,and possible molecular mechanism. Methods Construct two lentiviral vectors which contain shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and NC sequence( HPV18E6-RNAi-LV, NC-GFP-LV), based on the transduction with HPV18E6-RNAi-LV and NC-GFP-LV into HeLa cell to interfere the expression of HPV18E6 oncogene and NC sequence,the expressions of mRNA and protein( including phosphating patem)of HPV18E6, STATI, PKR, eIF2α, NF-κBp65, MAPK, JNK are measured with RT-PCR and Western blot, the difference of proliferation and sensitivity to carboplatin of HeLa cell are determined with Transwell cell methods and MTT among every groups. Results The expression of HPV18E6 oncogene can affect the expression level of mRNA and protein of NF-κBp65 and PKR genes, also affect phosphating levels of phosphating protein p-STAT1, p-PKR and p-eIF2α;the restraining rates of proliferation and sensitivity to carboplatin of HeLa cell are higher in HPV18E6-RNAi-LV group than the other groups( P＜0. 05 or P＜0.01 ). Conclusion HPV18E6 oncoprotein not only reduces the expression of PKR but dephosphorylates p-STAT1, pPKR and p-eIF2α to restrain activation of PKR/eIF2α signal transduction passage, maintain the proliferation and invading ability of HeLa cell and restrain apoptosis. The signal transduction among HPV18E6, MAPK/JNK are not clear.

Objective:To search for a rapid and efficient method for the isolation of 3 kinds of isoflavone glucosides from the ethanol extract of Semen Sojae Praeparatum.Methods: The crude isoflavones were extracted by 75% aqueous ethanol after removing the oil by Soxhlet extraction.Then 1300 macroporous resin,water and different concentrations of aqueous ethanol(10%,20%,30%,40%,50%,70%,and 95%) were used to separate and elute the crude isoflavones.The fraction eluted by 40% aqueous ethanol was subjected to preparative HPLC analysis.The chromatographic conditions: A YWG C18(10.0 mm?200 mm,i.d.10 ?m) column was used;the mobile phase consisted of acetonitrile-water-acetic acid at volume ratio of 25751(V/V/V) and a flow rate of 3.0 ml/min;the injection volume was 750 ?l;and the detection wave length was set at 260 nm.Results: Daidzin,glycitin and genistin were rapidly separated with the purities over 99% as determined by external standard HPLC.Conclusion: This technique is simple and suitable for the isolation of daidzin,glycitin and genistin from Semen Sojae Praeparatum.

Objective To study changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing (ERCC) 1 gene in ovarian cancer cell lines. Methods The small interference RNA (siRNA) targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to ovarian cancer cell line ES-2. The mRNA and protein of ERCC1 were evaluated by means of RT-PCR, western blot and immunocytochemistry. The changes of cisplatin sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTT) assay. Results In ES-2 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfection. The sensitivity to cisplatin of ES-2 cell line was increased by 53.88 times after disturbing the ERCC1 gene. Conclusion The sensitivity to cisplatin of ovarian cancer cell lines ES-2 could be enhanced by RNA interfering ERCC1 gene.

Objective To compare the esophageal acid and alkaline exposure characteristics of patients with cardia carcinoma after proximal gastrectomy or total gastrectomy.Method A total of 77 patients of cardia carcinoma who underwent radical resection from Sep 2007 to Sep 2011 in our hospital were retrospectively reviewed.24 hour esophageal pH monitoring were performed in all patients.Result Patients were divided into three groups:group TG had total gastrectomy (n =25),group PP had proximal gastrectomy with pyloroplasty (n =33),group NP had proximal gastrectomy (n =19).It revealed that indicators of acid reflux including the overall time length of acid episodes,＞ 5 min times of acid episodes,duration of longest acid episodes,time length of pH ＜ 4.00 and the DeMeester Scores in group NP were significantly higher than in group PP(U =32,P ＜ 0.01 ; U =35,P ＜ 0.01 ; U =23,P ＜ 0.01 ; U =39,P ＜0.01 ;U =49,P ＜0.01 respectively).Only alkline reflux was observed in group TG.The total times of alkaline episodes in PP group was significantly lower than in group TG(U =52,P ＜0.01) and group NP (U =182,P ＜0.01).＞5 min times of alkaline episodes in group TG was larger than in group PP,and that in group PP was larger than in group NP(P ＜0.01).Duration of longest alkaline episodes and total period of pH ＞7.00 in group PP was significantly higher than in group TG(U =125,P ＜ 0.01 ; U =143.5,P ＜ 0.01),and that in group TG was higher than in group NP(U =23.5,P ＜ 0.01 ; U =14,P ＜ 0.01).Conclusions Alkaline reflux deserves more attention in evaluating esophageal reflux in patients with cardia carcinoma after resection.Pyloroplasty is not helpful to relieving esophageal acid episodes while causing severe alkaline reflux.

Objective To study the changes of DNA repair genes and enhanced anti-tumor effect of cisplatin induced by mifepristone in human ovarian cancer drug resistance cells.Methods The alterations of cisplatin concentration producing 50％inhibition(IC50)in the COC1/DDP cell lines were examined by methyl thiazolyl tetrazolium(MTT)assay.RT-PCR and flow cytometry were used to analyze the changes of the mRNA of ERCC1,BRCA1,hMLH1 genes and cell cycle and apoptosis.Subcutaneous implantation of COC1/DDP was established in nude mice and the enhanced anti-tumor effect of cisplatin by mifepristone was observed in vivo.ResultsCisplatin IC50 values of COC1/DDP cell were decreased from(3.71±0.38)μg/ml to(3.18±0.46),(1.95±0.14),(0.64±0,18)μg/ml respectively when treated with 2.5,5.0,10.0 μmol/L mifepristone.Mifepristone could down-regulate the mRNA levels of ERCC1,BRCA1,hMLH1 genes and enhance G0/G1 phase block effect pf cisplatin,and 2.5,5.0,10.0 μmol/L mifepristone combined with cisplatin increased rate of cell apoptosis from 0.08％to 5.11％,9.13％and 12.24％ respectively.The percentage of inhibition of xenograft tumor volume in combined treatment group was 70.1％,which was significantly different(P＜0.05).Conclusion By down-regulating ERCC1,BRCA1,hMLH1 genes,blocking G0/G1 phase,and increasing apoptosis rate,mifepristone could enhance anti-tumor effect of cisplatin.

Objective:Human sera ACA was detected by ELISA method and some factors including the dilution rate of sera, blocking materials, temperature,ionic strength and interval time between blocking and detection,which could probablly influence the detective level of sera ACA were investigated.Methods:Human sera ACA was detected by ELISA.Results:10% NBS/PBS was more feasible than 1% bovine albumin/PBS for blocking non specific binding;the ACA titer of the same sample detected at 37℃ was lower than that of 22℃ and 25℃,the dilution rate 1∶50 for sera was suitable for the detection; time internal between blocking and detection could influence ACA level when it exceeding 2 weeks;the blinding ability of Ag with Ab was signicantly inhibited when the NaCl inonic strength was higher than 1.5 mol/L.Conclusion:The detected results of human sera ACA influenced by the dilution rate of sera,blocking materials,temperature ionic strength and internal time between blocking and detection.

OBJECTIVETo analyze the chemical constituents of supercritical carbon dioxide extraction products from Cnidium monieri.

METHODFour-factor and three-level orthogonal experimental design was used to optimize the SFE conditions as guided by the content of total coumarins in the extract. The chemical constituents were separated and identified by recrystalization.

RESULTOptimum extraction process was established: 25 MPa as extraction pressure, 50 degrees C as extraction temperature, 6.5 MPa as separation pressure and 60 degrees C as separation temperature.

CONCLUSIONChanges in extraction pressure, temperature, time, pulverized degree and separation pressure affect the extracting results remarkably. The two kinds of chemical constituents were separated by recrystallization from C. monieri and identified by the methods of UV, IR, MS, NMR.

Chromatography, Supercritical Fluid ; Cnidium ; chemistry ; Coumarins ; chemistry ; isolation & purification ; Fruit ; chemistry ; Furocoumarins ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

Studying the literature of blood stasis syndrome (BSS), we reviewed the research course and the perspective of molecular regulation network and BSS. The essence study of BSS was firstly proposed by Chen Ke-ji and Wang Jie, and developed for more than thirty years. The course for BSS study mainly included the formulation of BSS diagnostic standard, the establishment of BSS animal model, pedigree methods, twins combined clinical epidemiological survey of BSS research, the four "zu" subjects combined molecular regulation network of BSS, signal transduction system network and BSS research, and so on. Along with a new sequencing approach in basic research, clinical diagnostics, and drug development, we are promising to see the whole gene network research of human diseases, such as metabolic disease, cancer, and etc. These achievements could provide a new way of thinking for further studying the essence of BSS.
Diagnosis, Differential ; Genomics ; Humans ; Medicine, Chinese Traditional ; methods