Objective To study the multicomponent in vivo dynamic process in Chuanxiong Rhizoma;To elaborate in vivo metabolic profiling. Methods HPLC was used to establish the fingerprint of aqueous extract of Chuanxiong Rhizoma, and multicomponent changes were detected at the same time. Closed-loop intestine method was used to study the multicomponent changes of oral administration of Chuanxiong Rhizoma after stomach-intestine-liver process. Results Totally 17 components were detected in the fingerprint of aqueous extract of Chuanxiong Rhizoma and they were basically stable in the digestive juice. For in vivo metabolism, 4 components were metabolized by intestinal flora;3 components were metabolized by liver;2 new components were the metabolites of intestinal flora;1 component was the metabolite of liver. Conclusion Multicomponent sequential metabolism and closed-loop intestine method were used to clarify that multicomponent metabolic profiling was feasible, and it could provide experimental basis for the metabolism of traditional Chinese medicine.

This study was aimed to determine the solubility an d oil-water partition coefficient of main active com-ponents in Ge-Gen Qin-Lian (GGQL) Tablets (puerarin, baicalin and berberine hydrochloride) in phosphate buffer solution of different pH values and under the background of many components. Solubility of puerarin, baicalin and berberine hydrochloride in different medium pH, and oil-water partition coefficient of the octanol-water and oc-tanol-buffer system were determined by HPLC method. The results showed that the solubility and oil-water partition coefficient of puerarin, baicalin and berberine hydrochloride were varied with the change of pH, and varied under the background of components. At pH 7.4, the solubility was the biggest;puerarin was 7.56 mg·mL-1;baicalin was 17.07 mg·mL-1; berberine hydrochloride was 3.57 mg·mL-1. Oil-water partition coefficient P of these components at pH 1.0 was bigger;puerarin was 0.420 (lgP=-0.38);baicalin was 10.783 (lgP=1.03);berberine hydrochloride was 0.267 (lgP=-0.57). It was concluded that lipid solubility of puerarin, baicalin and berberine hydrochloride at pH 1.0 was better. It was speculated that better absorption in the stomach, and low lipid solubility under other pH. It was speculated that lipid solubility may be one of the reasons affecting the intestinal absorption.

Objective To select the components for quality control of Fructus Lycii based on the absorption of its extract. Methods To investigate metabolism of components of Fructus Lycii, everted rat gut sacs was carried out as well as the blood was taken from abdominal aorta,.and all samples were analysised by HPLC. Results There are twelve constituents absorbed between ileum and jejunum of rat , and four constituents were detected in the blood. Compound 2, 3, 4, 5, 6, 9, 10, 11 were absorbed in prototype forms in the intestine directly,and compound 1, 7, 8, 12 were new ones. On the other hand, four compositions(3, 7, 10, 13)could be absorbed into blood through analysis serum samples obtaining from aorta abdominalis of rats. Two of them (3, 10)could be absorbed directly by intestine, while(7)was absorbed into blood in new form . Conclusion Based on the intestinal absorption experiment and analysion of compsition in blood, components (3, 10, 13) can be the quality control ingredients of Fructus Lycii.

This study examined the changes of activities of vitamin K-dependent clotting factors (VKDCF) under various pathological conditions and explored the relationship between acquired deficiency of VKDCFs and hemorrhage. Clinical data of 35 patients who were diagnosed as having acquired deficiency of VKDCF were retrospectively analyzed. Coagulation factors involved in the intrinsic and extrinsic pathways were detected in these patients and 41 control subjects. The results showed that the average activities of VKDCFs were decreased in the patients in comparison to the control subjects and significantly increased after treatment of these patients with vitamin K and blood products. Multivariate regression analysis indicated that decreased activity of VKDCF was not an independent risk factor for bleeding disorders owing to deficiency or metabolic disturbance of vitamin K. It was concluded that acquired deficiency of VKDCF occurs under a variety of pathologic conditions and is closely associated with hemorrhagic events. Administration of vitamin K and transfusion of blood products containing high concentrations of VKDCFs helps alleviate the hemorrhagic diseases.

This study examined the changes of activities of vitamin K-dependent clotting factors (VKDCF)under various pathological conditions and explored the relationship between acquired deficiency of VKDCFs and hemorrhage.Clinical data of 35 patients who were diagnosed as having acquired deficiency of VKDCF were retrospectively analyzed.Coagulation factors involved in the intrinsic and extrinsic pathways were detected in these patients and 41 control subjects.The results showed that the average activities of VKDCFs were decreased in the patients in comparison to the control subjects and significantly increased after treatment of these patients with vitamin K and blood products.Multivariate regression analysis indicated that decreased activity of VKDCF was not an independent risk factor for bleeding disorders owing to deficiency or metabolic disturbance of vitamin K.It was concluded that acquired deficiency of VKDCF occurs under a variety of pathologic conditions and is closely associated with hemorrhagic events.Administration of vitamin K and transfusion of blood products containing high concentrations of VKDCFs helps alleviate the hemorrhagic diseases.

Objective To investigate the adhesion molecule expression and functional status of platelets in lung cancer patients, and their relations with disease progression. Methods Using flow cytometry to measure the expression of surface antigens and functional status of platelets in 60 healthy control group, and 164 lung caneer patients. Results Comparing with control group, the expression in early group and mid term group(A group) of CD31, CD36, CD62, CD63 increased, and there is no significant meaning in the differences of surgery group (B group) of CD31, TSP, CD36, CD62, CD63. The expression in advanced group (C group) of CD31, TSP, CD36, CD62, CD63 increased. Comparing with A Group, the expression in B Group of CD36, CD62, CD63 decreased. The expression in C group of CD31, TSP, CD36, CD62, CD63 increased. Comparing with the small cell lung cancer, the expression of adenocareinorna of TSP, CD36,, CD62, CD63, squamous cell carcinoma of CD31, CD62, CD63, and alveolus cancer of CD31, TSP, CD63, all decreased. Conclusion The high level expression of platelet activation exists in patients with lung cancer of different stage, and decreased after operation. Platelet activation expressed significantly in the advanced stage of small cell lung cancer.

Objective To evaluate the significance of TF-PCA to the diagosis of DIC by observing changes of tissue factor(TF) and procoagulant activity (PCA) in patients with disseminated inravaascular coagulation(DIC)according to the enhancenment degree of whole blood leukocyte-derived TF expression and TF-PCA stimulated by Lipopolysaccharide (LPS). Method Patients with acute leukemia (AL) were included during hospitalization from Jan. 2005 to Jan. 2007 in Union Hospital of Huazhong University of Science and Technology. Recalcification time of LPS-stimulated whole blood was applied to evaluate tissue factor clotting time (TiFaCT) : anticoagulated whole blood was incubated at 37°C for a certain time with or without LlS-stimulation, and then the recalcitication time was measured. TF-PCA were evaluated based on the decreased degree of whole blood mcalcification time(△s).LSI)- t test and bivariate correlation analysis were analyzed by using the SPSS software package (version 13.0 for Windows). A P-value less than 0.05 was considered statistically significant. Results A retrospective and contrast analysis indicated that △s in patients with DIC and patients suspected with DIC were (95.2±68.6) and (85.8±16.9), respectively. When compared with normal controls(30.4±25.1 ), the difference both had extremely statistical significe(P<0.01). The results of TF mRNA detection and TF-PCA inhibitory experiments showed that the method of TiFaCT had a high sersitivity and specificity for determination of TF-PCA. Condusiots The levels of TF-PCA were obviously elevated after stimulated by LPS in patients with DIC or suspected with DIC.TiFaCT has an important clinical reference value for the diagnostic and prognostic evaluation of DIC.

Objective To observe the changes of coagulation function in patients with acute lowerlimb deep venous thrombosis(DVT)and evaluate the risk factors for DVT. Methods Plasma APTT.PT,TT,D-dimer and fibrinogen(Fbg)were detected by an automated coagulation analyzer in 62 acute lower-limb DVT patients and 70 controls:Retrospective studies on the clinical data of all patients were done by binary logistic regression analysis.Results (1)In DVT group,plasma APTT,PT and TT,the levels of D-dimer and fibrinogen.and D-dimer/fibrinogen ratio(D/F ratio)were higher when compared with control group(au P＜0.01);(2)There were positive correlations between D-dimer and fibrinogen both in DVT and control groups(r=0.475,P＜0.01;r=0.564,P＜0.01,respectively);(3)Logistic analysis indicated that acute lower-limb DVT was associated with the presence of hypertension and increased plasma level of fibrinogen(OR=24.99,P＜0.01: OR=4.346.P＜0.01,respectively).Conclusions Hypertension and elevated plasma level of fibrinogen are independent risk factors for acute lower-limb DVT.

The effects of tissue factor (TF) on doxorubicin-induced apoptosis in human neuroblas- toma were investigated. The expression of TF was examined by Western blotting. TFsiRNA-pSUPER plasmid was constructed by inserting specific 19-nt silencing sequence targeting TF gene into pSU- PER vector. Transfection of TFsiRNA-pSUPER was performed using lipofectamine2000. The cytotox- icity of doxorubicin was determined by WST assay. The activation of Caspase-3 and PARP induced by doxorubicin was tested by Western blotting. The apoptotic cells were stained by Hochest33342 and counted under fluorescence inverted microscope. It was found that human neuroblastoma cell line SK-N-MC expressed high level of TF. Knockdown of the TF expression was achieved by trans- fection of TFsiRNA-pSUPER on SK-N-MC cells in a dose-dependent manner. Inhibition of TF sig- nificantly decreased the viability of transfected SK-N-MC cells treated with different concentrations of doxorubicin. Cleavage of Caspase-3 and PARP was enhanced in transfected SK-N-MC cells with down-regulation of TF. TFsiRNA treatment significantly increased the number of apoptotic cells in transfected SK-N-MC cells as compared with those control cells (P＜0.05) when these cells were ex-posed to 1 μg/mL doxorubicin for 8 h. These results suggested that knockdown of the TF expression by specific siRNA vector could increase the cytotoxicity of doxorubicin and enhance doxorubi- cin-induced apoptosis in human nearoblastoma cells. Over-expression of TF might contribute to chemotherapy resistance in human neuroblastoma and its progression, at lest in part, by regulating doxorubicin-induced apoptosis.

To clarify the role of TAFI in hypertensive disorders in pregnancy, 22 subjects, including 10 with pre-eclampsia (PE) and 12 with gestational hypertension were examined for the levels of TAFI and thrombin-antithrombin (TAT) complex. Thirty normal pregnant women served as controls. ELISA was employed for the detection. The results showed that the TAFI antigen levels in normal pregnancy group, gestational hypertension group and PE group were (85.35±24.69)%, (99.65±18.27)%, (110.12±23.36)%; (97.06±21.40)%, (114.08±27.76)%, (125.49±24.70)%; (106.6±19.21)%, (129.2±25.07)%, (139.1±30.12)%, in the 1st, 2nd and 3rd trimester respectively. No significant differences were found between the normal pregnancy group and gestational hypertension group but significant difference existed between normal pregnancy group and PE group in each tri- mester (P<0.05). TAT complexes were significantly higher in patients with PE than that in controls (P<0.05), but no correlation was found between TAT and TAFI. It is concluded that TAFI may con- tributed to the impairment of fibrinolysis in the patients with PE and may serves as a sensitive indi- cator for PE, but it may not help in the diagnosis of the gestational hypertension.