Objective To investigate the role of Akt signal pathway on the expression of monocyte chemoattractant protein-1 ( MCP-1 ) and nuclear transcription factor-κB (NF-κB) in renal tubular epithelial cells (HK-2) stimulated by albumin and to explore the mechanisms of action. Method The HK-2 cells were incubated in the presence of albumin (5,15,30 mg/mL) with or without Ly294002 (an inhibitor of Akt). Expression of mRNA was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).Expression of Akt and protein MCP-1 were assessed by Western blot. Electrophoretic mobility shift assay (EMSA) was used to detect the activation of NF-κB. q-test was used to evaluate the differences in means between groups. Results Compared with control group, the expression of MCP-1 mRNA remarkly increased. [Control group: 0.233 ±0.01; BSA(5 mg/mL) group: 0.285 ±0.04; BSA( 15 mg/mL) group:0.387 ± 0.02; BSA ( 30 mg/mL) group: 0.473 ± 0.05; BSA ( 30 mg/mL) + Ly294002 group: 0. 325 ±0.05, P ＜ 0.05 ]. The expression of MCP-1 protein in renal interstitum of operation group were remarkly increased too. [ Control group: 100 ± 15.1; BSA ( 5 mg/mL) group: 148 ± 19.3; BSA ( 15 mg/mL) group: 176±20.7; BSA(30 mg/mL) group: 263 ± 18.1; BSA(30 mg/mL) + Ly294002 group: 175 ± 18.0, P ＜0.05 ]. Albumin stimulated the expression of MCP-1mRNA and protein in a dose-dependent manner. Albumin remarkably increased the activity of NF-κB. Albumin enhanced the expression of Akt. Ly294002 inhibited albumin-induced the expression of NF-κB and partially decreased the level of MCP-1. Apositive correlation was noted between NF-κB activation and MCP-1 expression( r = 0.68 ,P ＜ 0.01 ). Conclusions Albumin-induces MCP-1 and NF-κB production via Akt signal pathway in renal tubular epithelial cells.

To establish the method of HPLC-fingerprint analysis for the quality control of Cnidium monnieri L. Cuss., and identify its active constituents by HPLC-MS, 35 batches of samples were analyzed on a Shimadzu C18 column with a gradient of acetonitrile and 0.1% aqueous aceticacid at a flow rate of 1.0 mL x min(-1) and detected at 245 nm and 322 nm. Furthermore, the typical samples were detected by HPLC-DAD-MS under negative ion mode. 35 batches of Cnidium monnieri L. Cuss. samples were classified into four types based on the results of similarity analysis. According to the comparison of the t(R), MS data and UV maximum absorbance (lambda(max)) values with the standards, 8, 7, 4 and 2 coumarins components were identified in four types of Cnidium monnieri L. Cuss. extracts, separately. The method is repeatable and reliable, and it is capable of effectively controlling the quality of Cnidium monnieri L.
Chromatography, High Pressure Liquid ; methods ; Cnidium ; chemistry ; Coumarins ; analysis ; Ecosystem ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Seeds ; chemistry ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Spectrophotometry, Ultraviolet

This study aimed to determine the impact of dentinal tubule orientation on dentin bond strength to provide a reference for clinical cavity preparation in resin-bonded restoration.Patients aged 13-16 years were selected,including 18 males and 21 females.Forty-eight human maxillary first premolars from orthodontic extractions were chosen to prepare the test models with the dentinal tubule orientations perpendicular and parallel to the bonding substrate.The test models in the vertical and parallel groups were divided into three groups:total-etching with 20％ phosphoric acid,total-etching with 35％ phosphoric acid and self-etching,with the dentinal tubule surfaces bonded with composite resin blocks in each group.After the standard test models of dentinal tubule-composite resin blocks were placed in distilled water and stored at 37℃ for 24 h,shearing tests were performed using a universal material testing machine at a crosshead speed of 0.5 mm/min.The bond strength values in the vertical group were 19.33±1.59 MPa for the 20％ phosphoric acid group,21.39±2.34 MPa for the 35％ phosphoric acid group,and 16.88±1.54 MPa for the self-etching group.The bond strength values in the parallel group were 24.53±1.99MPa for the 20％ phosphoric acid group,25.16±2.88 MPa for the 35％ phosphoric acid group,and 20.83±1.99 for the self-etching group.After using same total-etching adhesive,the shear bond strength of the parallel group was higher than that of the vertical group,and the difference was statistically significant (P＜0.05).Regardless of vertical group or parallel group,the difference in the bond strength value between the total-etching groups and the self-etching group was statistically significant (P＜0.05).It was concluded that the dentin bonding substrate which was parallel to the direction of the dentin tubule achieved an improved bond strength;the total-etching adhesives achieved higher bond strengths in dentin bond than the self-etching adhesives.

Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

ObjectiveTo determine the correlation between qualities of life(QOL) and depression,anxiety in cancer patients.Methods91 cancer patients was measured with self anxiety scale(SAS),self depression scale(SDS) and SF-36.ResultsDepression occurred in about 51.5% and anxiety occurred in about 24.1% of the general patients.The patients without depression showed a higher QOL than those with depression(P<0.05),and the patients without anxiety showed a higher QOL than those with anxiety(P<0.01).ConclusionDepression and anxiety are associated with poor quality of life,which should be put into plans of nursing.

Using the absorbent resin, silica gel and ODS column chromatography as well as semi-preparative HPLC, ten compounds were isolated from 70% ethanol extract of tubers of Dioscorea zingiberensis C. H. Wright, and their structures were elucidated as trigoneoside XIIIa (1), parvifloside (2), trigoneoside IVa (3), deltoside (4), protobioside (5), lilioglycoside k (6), zingiberensis newsaponin I (7), deltonin (8), prosapogenin A of dioscin (9), and trillin (10) on the basis of NMR and MS spectral data analysis. Among these compounds, 1, 3, 5 and 6 were isolated from this plant for the first time. In the screening test on platelet aggregation, compounds 7 and 8 exhibited induction effect on platelet aggregation, while compound 9 exhibited significant inhibitory effect on platelet aggregation in vitro.
Animals ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rats ; Rats, Wistar ; Saponins ; chemistry ; pharmacology

OBJECTIVETo study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection.

METHODSHEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P < 0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P < 0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group.

CONCLUSIONSHCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.

Cell Cycle ; Cell Cycle Proteins ; genetics ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; Embryo, Mammalian ; cytology ; Fibroblasts ; cytology ; metabolism ; Humans ; Lung ; cytology ; metabolism ; RNA, Messenger ; analysis

OBJECTIVETo estimate the size of female sex workers and clients in Taizhou city.

METHODSA household survey using network scale-up method (NSUM) was conducted among the 3000 community residents in Taizhou city from August to October in 2011, which aimed to estimate the social network size (c value) of Taizhou residents, and the c value was adjusted by demographic characteristics, back estimation and outlier elimination. Using the adjusted c value, the number of acquaintance of female sex workers or clients and the respect level toward female sex workers or clients were used to estimate the size of female sex workers and clients.

RESULTSA total of 2783 valid questionnaires were collected, among which 1380 (49.6%) were collected from Taixing city, 1403 (50.4%) were collected from Jingjiang city. 1334 respondents were male (47.9%) and 1449 (47.9%) respondents were female. The mean age was (39.4 ± 10.7) years. The average personal social network size using original data for Taizhou residents was 525, which differed from place, sex, age, educational level and marriage status. Using the remaining known populations through back estimation, the social network size was 419 and became 424 after the elimination of outliers. The estimated population size for female sex worker was 6370 (95%CI: 5886 - 6853), which accounted for 0.52% (6370/1 229 980) of the total number of female aged from 15 to 49. The estimated population size for clients was 15 202 (95%CI: 14 560 - 15 847), which accounted for 1.28% (15 202/1 190 340) of the total number of males aged from 15 to 49 and the ration of clients to female sex worker was 2.39:1.

CONCLUSIONNSUM is an easy and quick way to estimate the size of female sex workers or clients, but the estimated sizes are subject to bias and error due to estimate effect and sample representativeness.

Adolescent ; Adult ; China ; Female ; Humans ; Male ; Middle Aged ; Sex Work ; statistics & numerical data ; Sex Workers ; statistics & numerical data ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo establish a cell model of secreted alkaline phosphatase (SEAP) co-controlled by HCV 5'NCR and NS3 serine protease in an effort to develop new antiviral agents.

METHODSThe fragments of HCV 5'NCR and NS3/4A-SEAP were amplified by PCR. They were fused into pBluescript SK+ to generate 5'NCR-NS3/4A-SEAP chimeric plasmid. The resulting chimeric gene was subcloned into HindIII/Bsu36 I site of pSEAP2-Control (a SEAP eukaryotic expression plasmid), to generate pNCR-NS3/4A-SEAP, in which the SEAP was fused in-frame to the downstream of NS4A/4B cleavage site. The SEAP activity in the culture media of transiently transfected cells was monitored quantitatively. The regulatory effect of HCV 5'NCR and NS3 serine protease on SEAP expression was measured by treatment of transfected cells with antisense oligodeoxynucleotide (ASODN) against HCV 5'NCR and TPCK, a irreversible serine protease inhibitor.

RESULTSThe SEAP activity in the culture media reached 80801+/-4794 RLU, and was significantly inhibited by 5 micromol/L, 10 micromol/L of ASODN (t=4.315, p<0.01; t=6.985, p<0.001) and 100 micromol/L of TPCK (t=6.949, P<0.001).

CONCLUSIONA cell model of SEAP co-controlled by HCV 5'NCR and NS3 serine protease has been successfully established. This might promote the screening of anti-viral drugs

Alkaline Phosphatase ; secretion ; Antiviral Agents ; Drug Evaluation, Preclinical ; Hepacivirus ; genetics ; Hepatocytes ; enzymology ; virology ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Serine Endopeptidases ; biosynthesis ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics

OBJECTIVETo examine the role of domain II of hepatitis C virus (HCV) 5' noncoding region (5' NCR) in its translation initiation activity.

METHODSThe fragment of HCV 5' NCR with deletions of 5'-proximal 118 nucleotides were amplified with PCR, then was used to substitute for the full length HCV 5' NCR of plasmid pCMVNCRluc, a firefly luciferase (Fluc) eukaryotic expression plasmid regulated by HCV 5' NCR, to generate recombinant plasmid pCNl-d3. pCMVNCRluc, pCNl-d2 (modified pCMVNCRluc with deletions of HCV 5' NCR ntl-43) and pCNl-d3 were transfected into HepG2 cells with liposome transfection protocol, respectively, and the relative luciferase activity was measured to analyze the regulatory effect of the truncated 5'NCR on Flue gene expression. Meanwhile the Flue mRNA levels were detected by RT-PCR.

RESULTSThe recombinant plasmid was successfully constructed. The Flue mRNA levels of the 3 plasmids were not significantly different (P > 0.05), and there were also no significant difference between the relative luciferase activity of plasmids pCNl-d2 and pCMVNCRluc (P > 0.05). However, that of pCNl-d3 was decreased significantly (P < 0.01, compared with pCNl-d2 or pCMVNCRluc).

CONCLUSIONStructural domain II of HCV 5' NCR plays an important role in its translation initiation activity.

5' Untranslated Regions ; Hep G2 Cells ; Hepacivirus ; chemistry ; genetics ; metabolism ; Hepatitis C ; Humans ; Nucleic Acid Conformation ; Peptide Chain Initiation, Translational ; RNA, Viral ; chemistry ; genetics