The stems and branches of Hypericum petiolulatum were extracted by alcohol and liquid-liquid extraction. Seven furofuran lignans were isolated from the ethyl acetate fraction of ethanol extract of H. petiolulatum by using silica gelchromatography, Sephadex LH-20 chromatography, medium-pressure liquid chromatography and preparative HPLC. Their structures were identified by the spectroscopic methods as pinoresinol (1), medioresinol (2), 8-acetoxypinoresinol (3), epipinoresinol (4), (+)-syringaresinol (5), (+)-1-hydroxysyringaresinol (6) and erythro-buddlenolE (7). All the isolates were firstly found in H. petiolulatum. In the bioassay, compound 7 showed remarkable antioxidative activity inhibiting Fe(+2)-cystine induced rat liver microsomal lipid peroxidation with inhibitory rate 38% at a concentration of 1 x 10(-6) mol · L(-1) (positive control Vit E with the inhibitory rate of 35% at the same concentration).
Animals ; Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hypericum ; chemistry ; Lipid Peroxidation ; drug effects ; Microsomes, Liver ; drug effects ; metabolism ; Molecular Structure ; Oxidative Stress ; drug effects ; Plant Stems ; chemistry ; Rats

Background Serum cystatin C levels can be used to predict morbidity and mortality in patients with cardiovascular disease. However, the clinical relevance of serum cystatin C levels in patients with hypertensive left ventricular hypertrophy (LVH) has rarely been investigated. We designed the present study to investigate whether serum cystatin C levels are associated with cardiac structural and functional alterations in hypertensive patients. Methods We enrolled 823 hypertensive patients and classified them into two groups:those with LVH (n=287) and those without LVH (n=536). All patients underwent echocardiography and serum cystatin C testing. We analyzed the relationship be-tween serum cystatin C levels and LVH. Results Serum cystatin C levels were higher in hypertensive patients with LVH than in those without LVH (P＜0.05). Using linear correlation analysis, we found a positive correlation between serum cystatin C levels and interven-tricular septal thickness (r=0.247, P＜0.01), posterior wall thickness (r=0.216, P＜0.01), and left ventricular weight index (r=0.347, P＜0.01). When analyzed by multiple linear regression, the positive correlations remained between serum cystatin C and interventricular septal thickness (β=0.167, P＜0.05), posterior wall thickness (β=0.187, P＜0.05), and left ventricular weight index (β=0.245, P＜0.01). Con-clusion Serum cystatin C concentration is an independent marker for hypertensive LVH.

OBJECTIVE To study the effect of Qing Yi Tang on acute pancreatitis(AP),especially AP complicated with endotoxemia and its possible mechanism.METHODS Fourty Wistar rats were divided into 5 groups including group A(the control group),group B(the AP group),group C(the AP group treated with Qing Yi Tang),group D(the AP group treated with LPS) and group E(the AP group treated with LPS + Qing Yi Tang).Pathological damage of pancreatic tissue was scored with HE staining.The mRNA expression of TNF-? was measured with semi-quantitative RT-PCR,and activation of NF-?B was detected with flow cytometry(FCM) assay.RESULTS It was shown in results that the expression of TNF-? mRNA,activation of NF-?B and pathological damage of the group B were all obviously higher than those of the group A.After treated with LPS which might promote the activation of NF-?B,there was seen the further rise of the activation of NF-?B,expression of TNF-? mRNA and pathological damage.When Qing Yi Tang intervention was applied,the activation of NF-?B and the expression of TNF-? mRNA could be remarkably relieved,so did the pathological damage of pancreas.CONCLUSIONS Qing Yi Tang may be applied to decrease activation of NF-?B and the expression of TNF-? so as to treat AP or AP with endotoxemia.

Objective To determine whether intermittence irradiation of single blue or white light have an adverse effect on the DNA of newborn infants with hyperbilirubinemia by examining the sister chromatid exchange(SCE)frequency of peripheral blood lymphocytes.Methods The frequency of SCE in lymphocytes of 40 icteric infants treated by different phototherapy(PT) methods was a nalyzed by sister chromatid differetance staining technique (SCD).The patients receiving PT were divided into three groups according to two methods of PT,group A:single blue light,20 cases; group B:single white light,20 cases.Results 1.In group A, there was no difference between the levels of SCE before and after therapy within 3 days;but after 4 days, the levels of SCE increased.2.Obvious changes were observed in group B,and the frequency of SCE increased after 1 day and increased significantly in a dose-dependant manner.3.After treatment, the SCE frequency of group B was higher than that of group A.Conclusions PT has mutagenic effect on newborns with hyperbilirubinemia. The effect of single white light on peripheral blood lymphocytes of neonates is more significant.

Objective To investigate the expression of miR-21 and PDCD4mRNA in colorectal cancer and their correlation with clinicopathological characteristics,to elucidate relationship between PDCD4and miR-21 in vivo.Methods Expression of RNA (including miR-21and PDCD4mRNA) and PDCD4 protein were detected respectively by quantitative real-time reverse transcriptase-PCR and SP immunohistochemical staining in 43 cases of colorectal carcinoma and corresponding normal mucous membrane tissue.Results In colorectal carcinoma,expression of miR-21 was higher than that of the control ( P ＜ 0.05 ),and expression of PDCD4mRNA lower than the control ( P ＜ 0.05 ).Expression of miR21 was associated with lymph node metastasis and clinical stages ( Ⅰ + Ⅱ,Ⅲ + Ⅳ ) ( P ＜ 0.05 ).On the other hand,no significant differences were observed regarding sex,tumor site,size,local invasion,distant metastases,clinical stages( Ⅰ,Ⅱ ) (P ＞ 0.05 ).Expression of PDCD4mRNA was associated with local invasion,clinical stages ( Ⅰ,Ⅱ ) ( P ＜ 0.05 ).While no significant differences were found concerning sex,site,size,lymph node metastasis,distant metastases,clinical stages( Ⅰ + Ⅱ,Ⅲ + Ⅳ ) ( P ＞ 0.05 ).miR21 levels was negatively correlated with PDCD4 expression including nuclear and nuclear and cytoplasmic put together (P ＜ 0.05 ),in contrast to PDCD4mRNA and PDCD4 expression in cytoplasmic ( P ＞ 0.05 ).Conclusions ( 1 ) Abnormal expression of PDCD4 mRNA and miR-21 correlate with prognosis in colorectal cancer.(2) miR-21 suppress translation of PDCD4mRNA by binding to the 3'-untranslated region (3'-UTR)of target PDCD4mRNA.

Objective To understand the distribution of Oncomelania hupensis snails in source areas of the east route of Southto-North Warter Diversion Project and evaluate the effects of the snails on the safety of water transfer.Methods The investigation of snail distribution was carried out in the source areas of the east route of South-to-North Warter Diversion Project every spring.The method of the random sample combined with environmental sample was used for the field investigation.The beach land in the stilling pool of Jiangdu Pumping Station was selected as a surveillance site to observe the snail spread.Results The areas of the snail habitats and infected snails were 256.11,184.55,164.92,121.16 hm~2 and 8.27,1.0,1.0,0 hm~2 respectively in the source areas of the east route of South-to-North Wafter Diversion Project from 2006 to 2009.The densities of living snails had a downtrend,too.Google Earth showed that the areas of snail habitats distributed mainly in the Jiajiang River and Mangdao River in the source areas.The snail habitats were detected in the beach land in the stilling pool of Jiangdu Pumping Station.The research results showed that the snail spread related to the wastes from the river of drawing water.Conclusion There are the risks of snail spread in the source areas of the east route of South-to-North Wafter Diversion Project,so that the long-term surveillance and control on the snails is very necessary.

AIMTo develop a sensitive and specific HPLC method for determination of trigonelline in rabbit plasma, and study the pharmacokinetics in rabbit.

METHODSAfter ig of fenugreek extract and i.v. of trigonelline in rabbit, the biological samples could be well purified after precipitation of protein with methanol and acetonitrile. Asahipak NH2P-50 column was used, the mobile phase consisted of acetonitrile-water (90:10) at a flow-rate of 1.2 mL.min-1, and detection wavelength was set at UV 265 nm. The column temperature is 30 degrees C.

RESULTSThe calibration curve was linear in the range from 0.98 mg.L-1 to 31.28 mg.L-1, with r = 0.9986, the detection limit of this method was 50 micrograms.L-1. The concentration-time curves of trigonelline in rabbits after ig and i.v. administration were shown to fit one-compartment and two-compartment open model, respectively. The main parameters after ig of fenugreek extract were as follow: T1/2(Ka) was 0.9 h, T1/2(Ke) was 2.2 h, V was 0.64 L.kg-1, AUC was 1.93 mg.min.L-1. The main parameters after i.v. of trigonelline were as follows: T1/2 alpha was 10.8 min, T1/2 beta was 44.0 min, K21 was 0.044 min-1, K10 was 0.026 min-1, K12 was 0.017 min-1, AUC was 931.0 mg.min.L-1.

CONCLUSIONTrigonelline showed a middle rate of absorption and fast rate of elimination in rabbit. Meanwhile, the method is simple, accurate, with a good reproducibility, and it provide a basic method for the investigation of trigonelline and fenugreek pharmacokinetics.

Alkaloids ; blood ; isolation & purification ; pharmacokinetics ; Animals ; Antineoplastic Agents, Phytogenic ; blood ; isolation & purification ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Female ; Male ; Plants, Medicinal ; chemistry ; Rabbits ; Seeds ; chemistry ; Trigonella ; chemistry

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

OBJECTIVETo investigate hypermethylation of promoter region of RASSF1A and its relationship with ovarian malignant epithelial tumors.

METHODSMethylation-specific PCR (MSP) was used to determine the hypermethylation of promoter region of ras association domain family 1 (RASSF1A) gene in 80 cases of ovarian malignant epithelial tumors.

RESULTSNo methylation of promoter region of RASSF1A gene was found in all 80 normal control tissues (0). Of 80 ovarian malignant epithelial tumors 42 were hypermethylated in promoter region of RASSF1A gene (52.5%). There was no statistically significant difference in the frequency of hypermethylation of RASSF1A gene among serious adenocarcinomas, mucinous adenocarcinomas and endometrioid adenocarcinomas (54.2%, 52.4% and 45.5%, respectively; P > 0.05). Hypermethylation of RASSF1A gene happened more often in tumors in stage III and IV (66.7% and 77.8%) than that in stage I and II (21.4% and 16.7%; P < 0.05). It was less frequently observed in well and moderately differentiated tumors (34.5% and 35.0%) than in poorly differentiated tumors (80.6%; P < 0.05).

CONCLUSIONHigh frequency of methylation of RASSF1A promoter exists in ovarian malignant epithelial tumors as a tumor suppressor gene, its suppressor activity may be abrogated by an epigenetic mechanism. Hypermethylation of RASSF1A promoter in patients with epithelial malignant ovarian tumors is related to clinical stage and histopathological grade. It indicates poor prognosis.

Carcinoma, Endometrioid ; genetics ; Cystadenocarcinoma, Mucinous ; genetics ; Cystadenocarcinoma, Serous ; genetics ; DNA Methylation ; Female ; Humans ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo investigate the expression of two major alternative transcripts of RASSF1 gene (RASSF1A and RASSF1C) in human primary ovarian cancers and their biological implication as a new tumor suppressor gene.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and laser capture microdissection (LCM) were used to determine mRNA expression of the two major alternative transcripts of RASSF1 gene (RASSF1A and RASSF1C) in 3 ovarian cancer cell lines and 80 cases of primary ovarian cancers.

RESULTSRASSF1A mRNA was undetectable in SK-OV-3 cell line. Expression of RASSF1A and RASSF1C in 80 primary ovarian cancers were 40.0% (32/80) and 91.3% (73/80) respectively. RASSF1A mRNA expression was detectable more frequently in stage I and II (71.4%, 10/14; 75.0%, 9/12) than in stage III and IV ovarian cancers (26.7%, 12/45; 14.1%, 1/9) (P < 0.05). The expression level was also higher in well and moderately differentiated tumor groups (58.6%, 17/29; 50.0%, 10/20) than in poorly differentiated tumor group (16.1%, 5/31) (P < 0.05).

CONCLUSIONThere is a preferential loss of RASSF1A expression in human ovarian cancers and its expression is correlated with the tumor stage and the degree of histological differentiation which, as a tumor suppressor gene, might play an important role in the tumorigenesis of human primary ovarian cancer.

Carcinoma, Endometrioid ; metabolism ; pathology ; Cell Line, Tumor ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; biosynthesis ; genetics