Aim This study was designed to investigate the effects of pravastatin,a potent 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor,on vascular smooth muscle cells (VSMCs) proliferation and leukocyte-endothelial cell adhesion induced by lysophosphatidylcholine (LPC).Methods Cultured VSMCs from rabbit thoracic aorta were incubated with various concentrations of LPC in the absence or presence of different concentrations of pravastatin. MTT was used to evaluate the proliferation of VSMCs. We determined the effects of LPC and pravastatin on neutrophil K562 adhesion to endothelial cells ECV304 by directly counting non-adhesive K562 cells.Results Incubation of VSMCs with LPC (1～10 ?mol?L -1) stimulated proliferation of VSMCs in a time- and dose-dependent manner,while pravastatin (0.3～1 mmol?L -1) treatment prevented the proliferation of VSMCs caused by LPC. Moreover, incubation of ECV304 with 3 ?mol ? L -1 LPC for 12 h significantly enhanced K562 cells adhesion to endothelial cells, whereas pretreatment with pravastatin reduced leukocyte-endothelial cell adhesion. Conclusion These results suggest that pravastatin can antagonize the effects of VSMCs proliferation and leukocyte-endothelial cell adhesion induced by LPC.

OBJECTIVE:To study the influential factors related to efficacy of Peach kernel-Rheum palmatum couplet medi-cines in TCM formula,and to reveal the general regularity of compatibility environment,common ratio,processing variety and dosage forms of P. kernel-R. palmatum couplet medicines. METHODS:Using Chinese Medical Prescription Selected Dictionary ed-ited by Peng Huairen as data source,142 formulas of P. kernel-R. palmatum couplet medicines were collected. By establishing data-base,compatibility types of P. kernel-R. palmatum couplet medicines,as well as common ratio,processed prodact,dosage form were classified statistically. The influential factors related to efficacy of P. kernel-R. palmatum couplet medicines with different pro-portions were summarized. RESULTS:The efficacy of P. kernel-R. palmatum couplet medicines could be divided into 6 aspects and 11 roles,including activating blood circulation to dissipate blood stasis(activating blood to relieve pain,promoting blood circula-tion to eliminate disease,activating blood to promote menstruation,breaking stagnant and eliminating blood stasis),eliminating carbuncle and detoxicating(cleaning intestine and clearing away the pathogenic heat of lung,eliminating carbuncle and expelling pus,eliminating sore and detoxicating),expelling the pathogenic heat to loosen the bowels,warming yang for dispelling cold,forti-fying the spleen and nourishing the stomach,relaxing tendon and activating blood. The compatibility environment of P. kernel-R. pal-matum couplet medicines were mainly compatible with TCM for activating qi to eliminate stasis,activating blood to promote menstru-ation,breaking stagnant and eliminating blood stasis,expelling the pathogenic heat to expel stasis. The ratio of P. kernel to R. palma-tum ranged 1 : 8-4 : 1,and the ratio ranged 1 : 8-3 : 1 when performing the role of actirating blood circalation to dissipate blood stasis. Common processed products were crude P. kernel and prepared R. palmatum. Common dosage forms were mainly decoction,pill and powder. CONCLUSIONS:Compatibility environment,ratio,processing varieties,dosage forms influence the effects of P. kernel-R. palmatum couplet medicines,especially compatibility environment.

The overall concept is the core theory of traditional Chinese medicine ( TCM ) diagnosis and treat-ment system. During the process of building TCM effect evaluation on coronary heart disease (CHD), with the defined connotation and extension of heart diseases , except the consideration on its clinical symptoms , TCM four diagnosis and various current modern cardiovascular disease diagnostic methods should be organically com-bined according to the original macro basis . The micro quantitative evaluation is given on therapeutic effect of disease treatment in order to construct the complete TCM effect evaluation system.

OBJECTIVE:To establish a method for the contents of gallic acid,catechin,sennosides B,aloe-emodin,rhein, emodin,chrysophanol,physcion,chrysophanol-1-O- glucoside and emodin-8-O- glucoside in Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma,Shu Rhei Radix et Rhizoma,Rhei Radix et Rhizoma tan,Cu Rhei Radix et Rhizoma,and analyze the differ-ences. METHODS:HPLC was performed on the column was Hypersil C18 with mobile phase of methanol- 0.2% acetic acid(gradi-ent elution)at a flow rate of 1.0 ml/min,the detection wavelength was 260 nm,column temperature was 25 ℃,injection volume was 10 μl. RESULTS:The linear range was 0.252 5-4.040 0 μg for gallic acid(r=0.999 6),0.600 0-9.600 0 μg for catechin(r=0.999 6),0.297 4-4.758 4 μg for sennosides B(r=0.999 9),0.001 8-0.028 8 μg for aloe-emodin(r=0.999 9),0.005 0-0.080 0 μg for rhein(r=0.999 9),0.019 0-0.304 0μg for emodin(r=0.999 8),0.380 2-6.083 2μg for chrysophanol(r=0.999 7),0.008 2-0.131 2μg for physcion(r=0.999 8),0.126 0-2.016 0 μg for chrysophanol-1-O-glucoside(r=0.999 6)and 0.111 3-1.780 8 μg for emo-din-8-O-glucoside (r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 96.17%-97.21%(RSD=1.67%,n=6),97.60%-100.54%(RSD=2.55%,n=6),99.45%-101.32%(RSD=1.63%,n=6), 95.31%-98.19%(RSD=2.42%,n=6),98.99%-100.35%(RSD=1.86%,n=6),98.95%-101.21%(RSD=2.17%,n=6), 99.81%-100.62%(RSD=1.66%,n=6),96.78%-98.52%(RSD=1.99%,n=6),97.80%-100.14%(RSD=3.32%,n=6) and 97.40%-101.24%(RSD=2.89%,n=6). Compared with Sheng Rhei Radix et Rhizoma,the contents of gallic acid,catechin,sen-nosides B and anthraquinones in Cu Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma and Rhei Radix et Rhizoma tan decreased. The contents of catechin,sennosides B and anthraquinones in Shu Rhei Radix et Rhizoma. Catechin,sennosides B,chrysopha-nol-1-O- glucoside,aloe-emodin and rhein were not detected in Dahuang tan. CONCLUSIONS:The method is simple with good precision,stability and reroducibility,and can be used for the simultaneous determination of 10 chemical components in processed products of Rhei Radix et Rhizoma;there were significant differences in contents of 10 chemical components in processed prod-ucts of Rhei Radix et Rhizoma.

Objective:To observe the protein and mRNA expressions of microtubule-associated protein 1 light chain 3 (LC3)B, P62 and Beclin1 in the liver of rats with chronic fluorosis, and to explore the role of autophagy in pathogenesis of liver injury induced by fluorosis.Methods:Using a group design, 54 SD rats were divided into 9 groups according to their weight (100 - 120 g) using a random number table method, each group with 6 rats, half male and half female. They were control group (NC group), low fluoride group (LF group), high fluoride group (HF group), NC + rapamycin (RAP) group, LF + RAP group, HF + RAP group, NC + chloroquine (CQ) group, LF + CQ group, and HF + CQ group. The NC group drank tap water (fluoride concentration was 0.5 mg/L), LF group drank fluoride water (fluoride concentration was 5.0 mg/L), HF group drank fluoride water (fluoride concentration was 50.0 mg/L); NC + RAP group, LF + RAP group and HF + RAP group were fed with corresponding drinking water, respectively, for 3 months, and then RAP (1.5 mg/kg) was intraperitoneally administered for 10 d; NC + CQ group, LF + CQ group and HF + CQ group were fed with corresponding drinking water, respectively, for 3 months, and then CQ (60 mg/kg) was intraperitoneally administered for 10 d. Bone and 24-hour urine samples of rats in each group were collected to detect the contents of bone fluoride and urine fluoride; liver histomorphological changes were observed through hematoxylineosin staining; protein and mRNA expressions of LC3B, P62 and Beclin1 in liver were detected by immunohistochemistry and real-time fluorescence quantitative PCR, respectively.Results:Compared with the NC group [(0.03 ± 0.00) mg/kg, (0.34 ± 0.08) mg/L], the contents of bone fluoride [(3.86 ± 0.08) mg/kg] and urine fluoride [(1.11 ± 0.16) mg/L] in HF group were higher ( P < 0.05). In the NC group, the lobule structure of liver tissue was clear, the hepatic cords were arranged in order, and the cell structure was normal. There were different degrees of hepatocyte edema in LF and HF groups. After intraperitoneal injection of RAP, compared with the corresponding fluoride group, the morphology of hepatocytes did not change significantly. After intraperitoneal injection of CQ, compared with the corresponding fluoride group, the liver cells showed obvious edema, and the degree of edema aggravated with the increase of fluoride concentration. Compared with the NC group, the protein expressions of LC3B and Beclin1 in HF group were higher ( P < 0.05), and the protein expression of P62 was lower ( P < 0.05). After intraperitoneal injection of RAP, the protein expressions of LC3B and P62 in LF + RAP group was lower than that in LF group ( P < 0.05); Compared with HF group, the protein expressions of LC3B and Beclin1 in HF + RAP group were lower ( P < 0.05). After intraperitoneal injection of CQ, protein expression of P62 in LF + CQ group was higher than that in LF group ( P < 0.05); Compared with HF group, protein expression of P62 in HF + CQ group was higher ( P < 0.05). Conclusions:Early (3 month) fluoride intake could promote autophagy and induce edema of hepatocytes in rats, and RAP had similar effects. CQ may induce liver injury by inhibiting autophagy of hepatocytes.

Objective To analyze the ultrasonic features of thyroid microcarcinoma (TMC) and the causes of misdiagnosis. Methods The ultrasonic features including shape, margin, echogenecity, microcalcification, vascularity and lymphadenopathy were analyzed retrospectively in 26 pathologically-proven TMC patients. Results In 26 cases, 11 cases were diagnosed correctly before operation (11/26, 42.31%), 12 cases were misdiagnosed (12/26, 46.15%) as adenoma or benign nodule, and 3 cases were missed diagnosed (3/26, 11.54%). Among the 23 cases detected on ultrasound, 21 cases were solid and hypoechoic (21/23, 91.30%);19 cases were ill-defined (19/23, 82.61%);12 cases were taller than wide in shape (12/23, 52.17%); 14 cases had microcalcification (14/23, 60.87%); 7 cases showed central or peripheral blood flow signals (7/23,30.43%) with arterial resistance index>0.70 in 3 lesions and<0.70 in 4 lesions. Conclusions Several ultrasonographic features are helpful in identiifcation of TMC, including hypo/iso-echogenecity, ill-deifned margin, taller-than-wide shape, microcalciifcation, arterial signals with high resistance index, and abnormal lymphadenopathy. Moreover, for cases with multiple lesions, to the potential co-existence of benign and malignant lesions should be considered.

Objective To investigate the role of Th9 cells in primary biliary cirrhosis and its clinical significance.Methods The percentages of Th9 cells in patients with PBC (n=38)and healthy controls (n=38)were measured by flow cytometry. Patients clinical data were collected and Mayo risk scores were calculated.Determined the levels of IL-9,PU-1 and TGF-βmRNA expressions and the protein levels of IL-9 in plasma.Further analyzed the correlation between Th9 cells and clinical parameters.Results Compared to healthy controls,Th9 cells percentage was higher in PBC patients and increased with dis-ease stages (t=27.29;P<0.01).IL-9,PU-1 and TGF-βmRNA expressions were increased compared with healthy controls (t=14.69,23.92,10.48;all P<0.01)and the protein level of IL-9 was also up-regulated (t=11.66;P<0.01).The clinical parameters (ALP,AST,ALT,GGT and TBIL)were higher than healthy controls (t=10.94,12.95,10.56,14.92,27.70;all P<0.01).Moreover,the percentage of Th9 cells were positively correlated to Mayo risk,ALT,AST,GGT and TBIL (P<0.05).Conclusion Th9 cells may be involved in the pathogenesis of PBC and related to the disease stage,which provides new clues for future immunotherapy.

Objective To observe the effect of Ad-bTERTp-HSV-TK/GCV system on malignant ascites of mice and probe into its mechanism of action.Methods The SX1 inbred strain mice were injected with H22 cell line of liver cancer and were divided into 4 groups at random.The mice in each group were given corresponding treatment after 48 hours.The production of ascites and survival period were evaluated. The apoptosis rates of tumor cells were detected by FCM.Morphological changes of tumor cells were studied by electromicroscope.Results Compared with other groups.Ad-hTERTp-HSV-TK/GCV Can obviously inhibit the production of ascites(P<0.01),prolong the survival period (P<0.01),and apoptosis rate in this group (27.12±2.12)% was significantly higher than that in other groups.No obvious side effect Was found during the treatment.Conclusion Ad-hTERTp-HSV-TK/GCV system Can inhibit production of ascites and prolong the survical period of mice by inducing apoptosis of hepatoma cells,which is a safe and feasible treatment for hepatoma therapy.

Objective To construct the recombinant adenovirus vector with hTERT-HSV-TK and observe the killing effect of Ad-hTERTp-HSV-TK/GCV system on hepatocellular carcinoma cells. Methods A recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK was constructed via homologous recombination which both shuttle plasmid pSU-Tp-TK and adenovirus backbone plasmid pBHGE3 transfected into the HEK293 packaging cells. Then the Ad-hTERTp-HSV-TK was amplified and purified through PCR. The activity of the HepG2 cells and the L-02 cells were tested by methyl thiazolyl terazolium (MTT) after they were transfected by the recombinant adenovirus of different multiplicities of infection (MOI) and then were added GCV of different conc.entration. Results The recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK were identified by PCR successfully. The viral titer was 1.5×1010 pfu/ml after amplification and purification. The HepG2 cells were targetedly suppressed by Ad-hTERTp-HSV-TK/GCV system. The survival rate of cells decreased gradually along with the increase of the MOI and the GCV' s concentration. Conclusion The recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK can inhibit the HepG2 cells significantly, but has not influence on the L-02 cells.

Objective To investigate the mechanisms of phagocytosis of virulent Leptospira by peritoneal macrophages of guinea pigs,andevaluatetheroleof innateimmuneinthepathogenesisof leptospirosis. Methods Peritoneal macrophages of guinea pigs were extracted. Three specific inhibitors ( microfilament inhibitor cytochalasin D,microtube inhibitor colchicine and PI3K signalling pathway inhibitor LY294002) were added respectively to the macrophages 1 h before the infection of virulent Leptospira interrogans serovar Lai type strain Lai in vitro.Meanwhile, control group without inhibitor was established.Phagocytosis was observed by laser scanning confocal microscopy and phagocytic rates were evaluated by flow cytometry 3 h after infection.ResultsThe phagocytic rates of control group, cytochalasin D group, colchicine group and LY294002 group were (38.98 ± 0.91)%,(23. 99 ± 1. 40) % ,(40.81±0.91)% and (39.64 ±3.56) %, respectively.The phagocytic rate of cytochalasin D group was significantly lower than that of control group (P < 0. 05), while those of colchicine group and LY294002 group were not significantly different from that of control group (P >0.05). ConclusionMicrofilaments play an important role in the phagocytosis of strain Lai by peritoneal macrophages,but the process is independent on PI3K signalling pathway,and microtubes play little part during the phagocytosis.