Objective SIV30 protein of simian immunodeficiency virus(SIV)was prepared by genetic engineering technique as an antigen diagnostic reagent, to establish an immune comb method for the specific detection of anti SIV IgG in monkey serum. Methods Recombinant expression plasmid of SIV SIV30 gene was constructed by prokaryotic expression vector pGEX-4T-1, and expressed in the competent BL21 cells. The recombinant protein was purified as a diagnostic antigen, and a standardized procedure for the detection of immune comb was established and applied for clinical detection. Results The optimum coating amount of antigen was 0.02 mg/mL. The prepared IC was able to specifically detect the positive serum of SIV. There was no cross reaction between the sera of other viruses. It showed a high specificity of the detection method. Sensitivity analysis showed that the SIV30 protein was able to detect 1:400 times diluted SIV positive sera. The result of stability and repeatability test(the same sample was repeated 3 times) showed that the coefficient of variation(CV)was less than 10%. The serum samples of 10 suspicious monkeys were detected by this method, showing a consistent rate of comb method and ELISA test result of 100%, Kappa =1.000. Conclusions SIV30 protein is expressed in prokaryotic cells. The immune comb is prepared,and is successfullyl applied in clinical examination. It shows that the method has a high sensitivity, strong specificity, good reproducibility and practicability.

Pre-eclampsia (PE) is a pregnancy disorder that is characterised by severe hypertension and increased risks of foetal and maternal mortality. The aetiology of PE not completely understood; however, maternal nutrition and oxidative stress play important roles in the development of hypertension. The treatment options for PE are currently limited to anti-hypertensive drugs. Punicalagin, a polyphenol present in pomegranate juice, has a range of bioactive properties. The effects of supplementation with punicalagin on angiogenesis and oxidative stress in pregnant rats with induced hypertension were investigated. The pregnant rats were randomly divided into five experimental groups (n=12 per group). Hypertension was induced using an oral dose of NG-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg/day) on days 14–19 of pregnancy. Punicalagin (25, 50 or 100 mg/kg) was given orally on days 14–21 of pregnancy. Punicalagin treatment at the tested doses significantly reduced diastolic, systolic, and mean arterial blood pressure in L-NAME treated rats from day 14. Punicalagin also restored angiogenic balance by increasing the expression of vascular endothelial growth factor and downregulating vascular endothelial growth factor receptor-1/fms-like tyrosine kinase-1. Punicalagin, significantly increased the placental nitric oxide levels as compared to PE group. The increased levels of oxidative stress in rats with PE were markedly decreased by treatment with punicalagin. Punicalagin at the tested doses markedly (p < 0.05) enhanced the placental antioxidant capacity in L-NAME-treated rats. The raised catalase activity observed following L-NAME induction was significantly (p < 0.05) and restored to normal activity levels in punicalagin treatment. Further, 100 mg dose of punicalagin exhibited higher protective effects as compared to lower doses of 25 and 50 mg. This study shows that supplementation with punicalagin decreased blood pressure and oxidative stress and restored angiogenic balance in pregnant rats with induced PE.
Animals ; Antihypertensive Agents ; Arterial Pressure ; Blood Pressure ; Catalase ; Female ; Hypertension ; Hypertension, Pregnancy-Induced* ; Maternal Mortality ; Models, Animal* ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Oxidative Stress* ; Pre-Eclampsia ; Pregnancy ; Punicaceae ; Rats* ; Tyrosine ; Vascular Endothelial Growth Factor A

OBJECTIVE:To establish the method for the determination of oleanolic acid and ursolic acid in different medicinal parts of Tibetan medicine Pteocephalus hookeri,and compare the differences among the different parts. METHODS:The contents of oleanolic acid and ursolic acid from different medicinal parts(whole plant,aerial part,underground part)of P. hookeri were de-termined by UPLC-PDA. The separation was performed on Acquity UPLC HSS T3 column(150 mm×2.1 mm,1.8 μm)with mobile phase consisted of methanol-0.1 mol/L ammonium acetate(88:12,V/V)at the flow rate of 0.2 mL/min. The detection wavelength was set at 210 nm,and column temperature was 30 ℃. The sample size was 5 μL. RESULTS:The linear ranges of oleanolic acid and ursolic acid were 10.65-1065 μg/mL (r=0.9996) and 18.8-1880 μg/mL (r=0.9994),separately. The recoveries were 96.95%(RSD=1.24%,n=9) and 98.12%(RSD=2.13%,n=9),separately. RSDs of precision,stability and reproducibility tests were all less than 3%. The contents of oleanolic acid and ursolic acid from different medicinal parts in P. hookeri were in de-scending order of aerial part>whole plant>underground part;the average total content of oleanolic acid and ursolic acid from whole plants was 0.35%,the aerial part reached 0.56% and underground part was 0.09%. CONCLUSIONS:The method is rapid, accurate and reproducible,and it is suitable for the content determination of oleanolic acid and ursolic acid in different medicinal parts of Tibetan medicine P. hookeri. The contents of oleanolic acid and ursolic acid from aerial part of P. hookeri are higher than whole plant and underground part. It is suggested to use aerial parts of medicine.

OBJECTIVETo investigate the biocompatibility and degradation rate of crosslinking sodium hyaluronate gel with different ratio of molecular weight, so as to choose the effective, safe and totally degraded hyaluronate gel for aesthetic injection.

METHODS(1) Compound colloid was formed by cross-linking the divinyl sulphone and sodium hyaluronate with different molecular weight (4 x 10(5), 8 x 10(5), 10 x 10(5), 12 x 10(5)). (2) Healthy level KM mice was randomly divided into two groups to receive hyaluronic acid gel or liquid injection. Each group was subdivided into three subgroup to receive hyaluronic acid with different molecular weight. The biocompatibility and degradation rate, of hyaluronate were observed at 7, 90, 180 days after injection. At the same time, different molecular weight of sodium hyaluronate gel is sealed or exposed respectively under the low temperature preservation to observe its natural degradation rate. (3) The most stable colloid was selected as aesthetic injector for volunteers to observe the aesthetic effect.

RESULTSThe sodium hyaluronate gel with molecular of 4 x 10(5) was completely degraded 90 days later. The sodium hyaluronate gel with molecular of 8 x 10(5) was completely degraded 180 days later. The sodium hyaluronate gel with molecular of 10 x 10(5) was degraded to 90.0% after 180 days. The sodium hyaluronate liquid can be degraded completely within 7 days. The colloid could be kept for at least 12 months when sealed under low temperature, but was totally degraded when exposed for I d. Sodium hyaluronate gel with molecular 10 x 10(5) was confirmed to be kept for at least 6 months in animal experiment and clinical trials.

CONCLUSIONSUnder the same condition of material ratio, the higher the molecular weight is, the lower the degradation rate is. But the liquidity of gel is not good for injection when molecular weight is too large. It suggests that Sodium hyaluronate gel with molecular 10 x 10(5) maybe the best choice in cosmetic injections.

Animals ; Cross-Linking Reagents ; administration & dosage ; chemistry ; Hyaluronic Acid ; administration & dosage ; chemistry ; Injections, Subcutaneous ; Mice ; Molecular Weight ; Random Allocation

Objective With various doses of lipopolysaccharide (LPS) intra-amniotic injection in pregnant rats to establish histologic chorioamnionitis (HCA) model,and to determine the effects of HCA on fetal lung maturation and development in preterm fetal rats.Methods Thirty pregnant Sprague-Dawley rats were randomly assigned to four groups.On 19 days of gestation,rats in three model groups (n=8) were given intra-amniotic injection of LPS 2,8,10 μg,respectively,and those in the only one control group (n=6)was given normal saline (NS) 40 μl.Fetal rats were taken out 48 hours after LPS injection.Placenta,fetal membrane,fetal lung and umbilical cord were collected for pathological examination.mRNAs of surfactant protein (SP)-A,B and C and keratinocyte growth factor (KGF) in lung were determined by quantitative real-time polymerase chain preaction.KGF protein expression in lung was measured by immunohistochemistry.Morphologic observation of lung was performed on postnatal day 3.R× C table Chi-square test,KruskalWallis test and analysis of variance were used as statistical methods.Results (1) Fetal rat mortality in control,LPS2,LPS8 and LPS10 group was 0.0％ (0/55),22.5％ (18/80),51.4％ (36/70) and 87.5％(35/40),respectively,which increased with increasing dose of LPS (x2=46.183,P＜0.005).(2) HCA,fetal lung inflammation and umbilical vasculitis were induced in LPS groups,and the severity was dosedependent (fetal lung inflammation:Hc=39.84,HCA:Hc=41.13,umbilical vasculitis:Hc=41.52,all P＜0.01).(3) The expression of SP-A,SP-B,SP-C and KGF mRNAs in the four groups (control,LPS2,8,10) had statistical differences (SP-A mRNA:0.99±0.28,2.48±0.34,1.09±0.31 and 0.09±0.09,F=78.051; SP-B mRNA:0.99±0.34,2.23±0.53,1.49±0.51 and 0.14±0.06,F=28.327; SP-C mRNA:1.20±0.39,2.00±0.20,1.04±0.37 and 0.12±0.16,F=39.546; KGF mRNA:0.97±0.19,2.18±0.61,0.93±0.09 and 0.21±0.11,F=37.544; all P＜0.01).All mRNA expressions in LPS2 group were higher than those in the control group and those in LPS10 group was lower (all P＜0.05).(4) The expression of KGF protein in LPS2 group was significantly higher than that in other groups (0.60±0.20 vs 0.28±0.12,0.37±0.22,0.24±0.12,F=17.280,all P＜0.01).(5) Alveolarization was significantly inhibited in LPS8 group on postnatal day 3,and less maturity of pulmonary tissue was observed.Conclusions Various doses of LPS intra-amniotic injection in rats could induce HCA,fetal lung inflammation and umbilical vasculitis with different degrees.Histological inflammation would be worse with increasing LPS dosage.Moderate inflammation could promote lung maturation without inducing bronchopulmonary dysplasia,and KGF may play a role in this process.

Objective To investigate and analyze the influencing factors of community nursing cognition and employment intention of higher vocational nursing students,and provide reference for higher vocational colleges to carry out community nursing education and employment guidance.Methods Using the stratified cluster sampling method,nursing students from different grades and different education levels were selected as the object of study and participate in the questionnaire survey.Results The nursing students from different grades,education levels and areas showed significant difference in community nursing and employment intention.Conclusions Higher vocational nursing colleges should strengthen the community nursing practice education and community employment guidance; improve the community nursing cognition and community employment intention of nursing students.

Objective To evaluate the expression of Krüppel-like factor 8 (KLF-8) and matrix metalloproteinase 9 (MMP-9) in placentas and their relationship with the pathogenesis of preeclampsia (PE).Methods Twenty-two women with PE (mild PE:4 cases; severe PE:18 cases) who received cesarean sections in the First Affiliated Hospital of Chongqing Medical University from September 2011 to March 2012 were recruited as the PE group (n =22).And twenty women who received elective term cesarean section without perinatal complications were chosen as the control group (n =20).Placentas were collected and immunohistochemical SP method were employed to detect the localization of KLF-8 protein.KLF-8 mRNA level was determined by quantitative real-time PCR technique and western blot analysis was used to quantify KLF-8 and MMP-9 protein levels.Results (1) There was no difference of KLF-8 protein distribution in placentas of the PE group and the control group.It was mainly located in the nuclear and cytoplasm of syncytiotrophoblasts.KLF-8 immunostaining was apparently decreased in the placentas of preeclamptic women when compared with the control group.(2)The KLF-8 mRNA levels were significantly decreased in placentas of the PE group (0.69 ±0.08) compared to those of the control group (1.14 ±0.09,P ＜0.01).(3) KLF-8 and MMP-9 protein levels significantly decreased in the PE placentas (0.68 ±0.05 and 0.21 ± 0.03) when compared to the control group (0.94 ± 0.06 and 0.34 ± 0.03,respectively,P ＜ 0.01).(4) There was a positive correlation between the expression of KLF-8 and MMP-9 protein in the placentas from PE and normal pregnancies (r =0.64,P ＜ 0.01).Conclusions KLF-8 mRNA and protein levels were decreased in placentas of PE patients compared to those of normotensive women.KLF-8 protein was primarily located in the invasion-related trophoblast cells and its expression had a positive correlation with MMP-9 levels.KLF-8 might have an important role in the pathogenesis of PE by regulation of trophoblast invasion.

ObjectiveTo evaluate the effectiveness and safety of high frequency electronic associating stents insertion and subsequent radiotherapy guided by fiberoptic bronchoscope in treating tracheobronchial stenosis caused by lung and esophagus carcinoma.MethodsFifty-two patients which were ascertained with serious stenosis in trachea or bronchi by fiberoptic bronchoscope were subjected to the study.Fiberoptic bronchoscope was used to investigate the location,extent,blood supply and the degree of tracheobronchial stenosis before the study.After the focus in the airways was cleared up with high frequency electronic,electrocoagulation and snare under the guide of the euthyphoria of fiberoptic bronchoscope,Ni-Ti memory alloy stents were placed into the stenosis airways.The 52 patients were divided into two groups with 26 cases each:the stent group and the stent associating radiotherapy group(radiotherapy group).The later group accepted subsequent radiotherapy after the therapy.The following indexes of the two groups were investigated:short-time clinical effect,dyspnea index class,rate and average time of airway restenosis,life span and survival rate.ResultsAfter therapy,the 52 patients had obvious improvements in dyspnea and the diameter of the stenosis airways.The short-time total effective rate was 100.0％ (52/52),but there was not significant difference between the two groups(P＞ 0.05 ).After therapy,there was significant difference in two groups in dyspnea index class(P ＜ 0.01 ).Although there was not significant difference in the rate of airway restenosis between the two groups(P＞0.05),the average time of airway restenosis in the radiotherapy group was obviously longer than that in the stent group(P＜ 0.01 ).The survival rate of the radiotherapy group in the 6th and 9th month after therapy was higher than that in the stent group (P ＜ 0.05),but there was no significantdifferenee in the 3rd and 12th month (P ＞0.05).ConclusionHigh frequency electronic associating stents insertion and subsequent radiotherapy guided by fiberoptic bronchoscope is an effective and safe treatment for the patients suffering tracheobronchial stenosis caused by lung and esophagus carcinoma.

OBJECTIVETo develop an identification method with significant specificity for patchouli oil.

METHODThe fingerprint was performed by gas chromatography with patchouli alcohol and pogostone as chemical markers.

RESULTThe similarity of 12 samples were higher than 0.9 and it can be used to identify the characteristics of patchouli oil.

CONCLUSIONThe GC fingerprint can be used for identification of patchouli oil.

Chromatography, Gas ; methods ; Lamiaceae ; chemistry ; Oils, Volatile ; analysis ; Plant Oils ; analysis ; Sesquiterpenes ; analysis

OBJECTIVETo isolate and identify chemical constituents of Cassia alata.

METHODCompounds were separated and purified by column chromatography with silica gel and Sephadex LH-20, elucidated by spectroscopic methods including 1D and 2D NMR, IR and MS techniques.

RESULTTwelve compounds were isolated from C. alata, which were identified as chrysoeriol (1), kaempferol (2), quercetin (3), 5,7,4'-trihydroflavanone (4), kaempferol-3-O-beta-D-glucopyranoside (5), kaempferol-3-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside (6), 17-hydrotetratriacontane (7), n-dotriacontanol (8), n-triacontanol (9), palmitic acid ceryl ester (10), stearic acid (11), palmitic acid (12).

CONCLUSIONCompounds 3-12 were isolated from C. alata for the first time.

Cassia ; chemistry ; Organic Chemicals ; analysis ; chemistry ; isolation & purification ; Plant Leaves ; chemistry