OBJECTIVE: To explore the pathogenic mechanism of liver-yang hyperactivity type of hypertension and to observe the effects of Pinggan Qianyang Formula (PGQYF), a compound of traditional Chinese herbals for calming the liver and suppressing yang, so as to provide experimental evidence for new marker proteins of drug therapy. METHODS: A rat model of liver-yang hyperactivity was prepared with spontaneous hypertensive rats (SHRs) by administration of Aconiti Praeparatae Decoction. Adrenal proteins were separated by 2D gel electrophoresis (2-DE). The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database analysis. RESULTS: The rat model of liver-yang hyperactivity was successfully reproduced, and the PGQYF could decrease the grades of irritability, conjunctival congestion and systolic blood pressure of the rats (P<0.05, P<0.01). After analysis, twelve obviously differentially expressed proteins were found, eight of which were identified. The expression levels of isocitrate dehydrogenase and steroidogenic acute regulatory protein in the untreated group were up-regulated as compared with those in the normal control group, and down-regulated in the treatment group. The expression levels of ferritin light chain, elongation factor Tu, Rho GDP disassociation inhibitor 1, flavin reductase and basic transcription factor 3 in the untreated group were down-regulated as compared with those in the normal control group, and up-regulated in the treatment group. CONCLUSION: Differentially expressed adrenal proteins in SHRs with live-yang hyperactivity are successfully identified. This approach may lay a foundation for the further investigation of pathogenic mechanisms in hypertension with liver-yang hyperactivity and the mechanisms of PGQYF treatment.

Objective To observe the effect of therapy of soothing liver and subsiding yang on hypothalamic protein expression in hypertensive rats with hyperactivity of liver yang,and to explore its therapeutic molecular mechanism for hypertension.Methods Spontaneous hypertensive rats (SHR)were given gastric gavage of Aconiti Praeparatae Decoction (APD)to induce hypertensive rat model with hyperactivity of liver yang,and SD rats served as the normal control.The treatment group received modified Tianma Gouteng Decoction (TGD,0.1g/mL).Two-dimension gel electrophoresis (2-DE)was used to separate the hypothalamic proteins,and the differential expressed proteins were identified by matrix-associated laser desorption-ionization time-of-flight massspectrometry (MALDI-TOF-MS)and database analysis.Results Hypertensive rat model with hyperactivity of liver yang was established successfully.The irritation degree,in-take water volume and systolic pressure were increased in the model group(P

Objective:To detect the expression of cyclin B 1 in human ameloblastoma(AB). Methods:Expression of cyclin B1 was detected in 73 cases of AB (32 primary, 33 recurrent and 8 malignant cases), 19 of odontogenic keratocyst (OKC) and 7 of normal oral mucosa by streptovidin-biotin method. Results:The ratio of positive nuclear expression of cyclin B 1 in normal oral mucosa, OKC and AB was 28.6%(2/7), 36.8%(7/19) and 63.0%(46/73) respectively (P

Objective: To investigate the effects of bovine plasma fibronectin on the proliferation and differentiation of rat osteoblasts. Methods:Rat osteoblasts were cultured and identified. AlamarBlue were used to determine the effect of various concentration of bovine plasma fibronectin on the proliferation of the osteoblasts. ELISA method was used to determine the alkaline phosphatase activity of the cells. Results: Fibronectin stimulated the proliferation of rat osteoblasts at 40 ?g/ml (P

Objective To observe the recent effects and toxicity in advanced gastric carcinoma by two kinds of neoadjuvant chemotherapies ( PF and FOLFOX).Methods A total of 60 patients with advanced gastric cancer ( Ⅲ ～ Ⅳ ),they were randomly divided into 2 groups; PF group ( 30 cases) and FOLFOX group ( 30 cases).All patients were given 3 cycles of docetaxel or L-OHP chemotherapy preoperatively,to observe the recent effects and toxicity in advanced gastric carcinoma.Results The total effective rate in PF group was 77.67％,while it was 50.00％ in FOLFOX group,it had significant difference between in two groups ( x2 =4.59,P ＜ 0.05 ).There is no difference in the incidence of neutropenia,transaminase elevation,nausea and diarrhea two groups.The incidence of hair loss in PF group is higher than in FOLFOX group,but there is no difference between two groups; The incidence of peripheral nerve paresthesia in FOLFOX group is higher than that in PF group( x2 =5.08,P ＜0.05).Conclusion PF as neoadjuvant chemotherapy is a very effective and well-tolerated regimen in patients with advanced gastric cancer.

Evidence-based medicine is advocated by WHO and adopted by developed countries for many years. In China, however, the selection of essential medicine and various medical insurance reimbursement schemes medicine is usually based on experts' experience of prescription practice which is under heavy critics resulting from the lack of related comparative efficacy and evidence-based research. The efficacy of Jian'ganle in prevention of drug-induced liver injury (DILI) caused by antituberculotics was evaluated in this study by comparison with Hugan Pian, glucuronolactone and reduced glutathione. Evidence was provided for relevant sectors such as Ministry for Human Resources and Social Security of the People's Republic of China and National Health and Family Planning Commission of the People's Republic of China to select and renew the Essential Medicine List (EML), the new rural cooperative medical scheme in China (NRCMS) list or the reimbursement list of industrial injury insurance. A total of 189 patients with initial pulmonary tuberculosis were divided into four groups who took antituberculotics combined with Jian'ganle, Hugan Pian, glucuronolactone and reduced glutathione respectively. Their liver function profile including alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL), total protein (TP), albumin (A) and globulin (G) were detected at admission as baseline and after treatment. The Jian'ganle group was compared with the three others by chi-square tests. In an aspect of maintaining bilirubin indexes normal, Jian'ganle was more efficacious than glucuronolactone. And Jian'ganle had a little more efficacy than reduced glutathione to maintain protein indexes normal as well. And the therapeutic regimen of antituberculotics combined with Jian'ganle was the best in treating tuberculosis and preventing DILI at the same time. The study showed that among the four hepatinicas which demonstrated similar prevention of DILI caused by antituberculotics, Jian'ganle has more advantages over the three others to some extent, which provides a reliable basis for health sectors to select and renew the EML, NRCMS List or the reimbursement list of industrial injury insurance.

To study the metabolite excretion of theophylline, a rapid and specific method by liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) method for simultaneous determination of theophylline, 1, 3-dimethyluric acid (1,3-DMU), 3-methylxanthine (3-MX) and 1-methyluric acid (1-MU) in human urine was developed using theophylline-d6 and 5-fluorouracil as internal standards. Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the negative mode for mass spectrometric detection. After diluted with methanol and centrifuged, the analytes and ISs were separated on a XDB-Phenyl (150 mm x 4.6 mm, 5 microm) column with a mixture of water-methanol-formic acid (30 : 70 : 0.15) as mobile phase at a flow rate of 0.6 mL x min(-1). The linear calibration curves for theophylline, 1, 3-DMU, 3-MX and 1-MU were obtained in the concentration range of 1.0-250 microg x mL(-1), separately. The method herein described is effective and convenient, and can be used for determination of theophylline and its three metabolites. The results showed that urinary excretion ratio of theophylline, 1,3-DMU, 3-MX and 1-MU is approximately 1 : 3 : 1 : 2 in Chinese subjects, which is similar to the reported excretion pattern in Caucasian.

Objective To investigate the inhibitory effect of extracts from asparagus filicinus rhizome on prolifieration of human osteosarcoma Saos-2 cells and its molecular mechanism .Methods MTT assay was used to detect the cytotoxic activity and growth inhibition of three different extracts from asparagus filicinus rhizome against Saos-2 cells ;plate colony formation assay was per-formed to detect active fraction of asparagus filicinus rhizome on the anchorage dependent growth of Saos-2 cells ;the cell cycle alter-ation was determined by propidium iodide staining and flow cytometry analysis ;the alteration of protein expression level of COX-2 was determined by using Western blotting .Results Ethyl acetate fraction of asparagus filicinus rhizome (AF-A) exerted the potent cytotoxicity on Saos-2 cells(IC50 =26 .7 μg/mL);AF-A induced the inhibitory effect on the anchorage dependent growth of Saos-2 cells in a dose dependent manner(P<0 .05);Saos-2 cells treated by AF-A at the concentration of 30 .0 and 100 .0 μg/mL for 48 h induced the increase of percentages of S phage from (31 .8 ± 4 .8)% in the control group to (43 .7 ± 2 .5)% and(51 .9 ± 1 .9)% ,the difference showing statistical significance (P< 0 .05) .Western blotting showed that AF-A at different concentrations decreased COX-2 protein expression .Conclusion AF-A posseses the inhibitory effect on the proliferation and growth of human osteosarcoma cells in vitro ,and its mechanism might be associated with the induction of S phage arrest and the inhibition of COX-2 protein ex-pression level .

Objective To investigate the relationship between the expression of the survivin gene and CpC methylation in exon 1 of the survivin gene in CA tissue, and to study the expression of survivin protein in CA tissue and its modulation mechanism. Methods Tissue samples were obtained from the CA lesions of 30 patients, normal cervix of 10 female controls, and normal foreskin of 10 male controls. Immunohistochemistry was carried out to detect the expression of survivin protein in these specimens, RT-PCR to measure the mRNA expression of survivin, and methylation specific PCR (MSP) to analyze the methylation status of CpG island in the survivin gene exon 1. Results The positivity rate of survivin protein and mRNA was 90% (27/30) and 93.3% (28/30) in CA tissue specimens, respectively, 5% (1/20) and 5% (1/20) in control tissue specimens, respectively; there was a significant difference between the two groups of specimens in both the parameters (x2 = 35.187, 38.437, both P < 0.01). The demethylation of CpG island in the survivin gene exon 1 was observed in 86.7% (26/30) of the CA tissue specimens and in 15% (3/20) of the control tissue specimens (x2 = 10.865, P < 0.01). There was a positive correlation between the demethylation status of CpG island in exon 1 and the mRNA expression of survivin gene (x2 = 13.929, P = 0.014). Conclusions The expression of survivin protein in CA tissues might be associated with the demethylation of CpG island in exon 1 of the survivin gene, and may play a certain role in the development of CA.

This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.