OBJECTIVE: To establishes an HPLC method for determination of the contents of Osthol and Berberine in Kangfuyan effervescent suppository. METHODS: HPLC was carried out on a column of Diamonsil (250 mm?4.6 mm,5 ?m) with mobile phase consisted of acetonitrile-0.2% phosphate acid (52∶48,Lauryl sulfonic acid sodium 0.1 g was contained in every 100 mL) with flow rate at 1 mL?min-1.The detection wavelength was set at 322 nm and the column temperature was set at room temperature. RESULTS: The linear range was 0.019 8～0.396 0 ?g(r=0.999 97) for Osthole versus 0.056 65～1.133 00 ?g(r=0.999 99) for Berberine. The average recoveries of Osthol and Berberine were 101.06% (RSD=1.10%) and 101.88%(RSD=0.98%), respectively. CONCLUSION: This method is simple, accurate and reproducible, and applicable for the quality control of Kangfuyan effervescent suppository.

Objective To observe the efficacy of extracorporeal liver support by using less fresh frozen plasma in the treat‐ment of acute‐on‐chronic liver failure.Methods A total of 45 patients with acute‐on‐chronic liver failure were divided into ob‐servation group[plasma perfusion(PP) with a small amount of plasma+ plasma exchange(PE)] ,control group 1(PE) ,control group 2(PP+PE)in terms of the amount of plasma used on the day of treatment. All the patients received artificial liver treatnts 62 times totally.Results The clinical symptoms were improved in the three groups after treatments.There were significant differences in the decrease of alanine transaminase (ALT) ,aspartate transaminase(AST) and direct bilirubin(DBil)rather than the decrease of total bilirubin(TBil)and blood ammonia among the groups.No significant difference was noted in the liver and kidney function among the three groups. The improvement of the coagulation function was poor in the observation group when compared with the control group 1 and control group 2 and there were significant differences.Conclusion During the short sup‐ply of the plasma ,plasma perfusion combined with small amount of plasma can be considered to be used in artificial liver treat‐ments ,which can effectively decrease the level of TBil ,relieve symptoms and decrease the occurrence of complications.

Objective To explore the percentage changes of peripheral T , B cell subsets and NK cells in chronic HBV infectors under different immune states and hepatitis B cirrhosis . Methods Seventy-five chronic HBV infectors, including 20 cases with immune clearance, 20 cases with immunodeficiency (inactive) and 35 cases with cirrhosis, and 20 healthy control were enrolled. The percentages of peripheral T and B lymphocyte subsets and NK cells were detected by Flow Cytometry. The differences of the groups were analyzed. Results Comparing with the control group, CD4+T cells were decreased in the other four groups (P<0.05). The sequence of CD4+T cells, from high to low, was the control group, the immunodeficiency group, the immune clearance group, the compensated cirrhosis group and the de-compensated cirrhosis group. CD4+/CD8+T cell and NK cell were lower , but CD8+T cell and B cell were higher in immune clearance group than that of the control group (P < 0.05). Patients in immunodeficiency group had lower ratio of CD4+/CD8+T cell and higher CD8+T cell than those in the control group (P < 0.05). In all the groups, patients with de-compensated cirrhosis showed highest ratio of CD4+ to CD8+ T cells and B cells, but lowest CD3+T, CD8+ T and NK cells (P < 0.05). Conclusions Results suggests immune dysfunction exists in patients with chronic HBV infection. It has potential clinical value in understanding patients′ immune states and progression of disease by detecting peripheral blood lymphocyte subsets and NK cells.

Objective To investigate the different immune status of TCM classification and the levels of T and B cells and the re-lationship between the NK cells .Methods Three different immune state of patients with chronic HBV were divided into asthenia syndrome(AS) group and sthenia syndrome(SS) group ,and then we detected the peripheral blood lymphocyte subsets and NK cells ,and compared the above each index analysis .Results In three different immune status ,all patients with AS and SS groups CD4+ T cell percentages are lower than normal control group (P<0 .05) ,and in a state of immune tolerance ,CD4+ T cells of SS group were lower than the AS group (P<0 .05) ,in a state of immune clearance ,CD8+ T cell percentage were SS group> control group> the AS group (P<0 .05) .Three different immune status ,the ratio of CD4+ /CD8+ SS group were significantly lower than the AS group or control group ,in a state of immune clearance the SS and the AS group B lymphocyte percentage were higher than the control group ,the percentage of NK cells was lower than the control group (P<0 .05) .Conclusion Chronic HBV infection have immune dysfunction ;HBV infected different immune status of TCM classification have relationship with T and B cell subsets and NK cells ,and the immune function test has certain clinical application value for the judgment of TCM syndrome type .

Objective To investigate the correlation and clinical diagnosis significance between ultrasound elastography and the distribution of myofibroblast in breast tumor.To assess the value of ultrasonic elastography and myofibroblast in the diagnosis of breast cancers.MethodsThree-hundred and fifteen patients recruited from May 2009 to November 2010 were divided into benign group and malignant group according to postoperative pathological diagnosis results considered as gold standard.The clinical value of the score of ultrasonic elastography was evaluated.The expression levels of CD34 and α-SMA protein in breast tissues were examined by immunohistochemistry.ResultsThe difference was statistically significant for the expression level of CD34 and α-SMA between malignant breast tumor patients compared with benign ( P ＜0.05).The expression level of CD34 was lower,but α-SMA was higher in the patients of breast cancer compared with benign tumor.However,the expression level of CD34 and α-SMA was just the opposite in the patients of benign tumor.The expression level of CD34 was the negative correlation with the elastography score ( P ＜0.05) in the breast tumor,but α SMA just the opposite situation.Conclusions The score of ultrasound elastography can represent MFS distribution characteristics in breast tumors in the result that α-SMA +/CD34- can determine the existence of myofibroblast.

Objective:To observe the changes of TLR4 and IL-6 expression in rats hippocampus CA1 region in remote ischemic postconditioning(RIP) and explore its significance.Methods: All the 72 male SD rats were divided into Sham group,Contrast group and RIP group randomly.Each group was divided into 4 time points:12h,24 h,48 h and 72 h group.There were 6 rats in each group.Use the cerebral ischemia-reperfusion model which was established with modified Longa method as the contrast group.The method of RIP was to Fasten rats Posterior limbs by a tourniquet for 30 min immediately,then relax them for 30 min, repeat 3 times.To observe the pathological variation of hippocampus CA1 region by HE dyeing;to test the expression of TLR4 and IL-6 by immunohistochemical staining.Results: Compared to contrast group, neuronal loss and swelling reduced significantly in RIP group.Compared to sham group, the TLR4 and IL-6 expression in contrast group and RIP group increased significantly ( P<0.05 ) .Compared to contrast group,the TLR4 and IL-6 expression in RIP group reduced significantly(P<0.05).Conclusion:RIP dose have protective effect on cerebral ischemia.The effect may be associated with the inhibition of TLR4 and IL-6 expression.

Objective:To elucidate the changes in apparent diffusion coefficients(ADC)by quantify diffusion weighted (DW)magnetic resonance imaging(MRI)in patients with Alzbeimer's disease(AD),and the relationship between micro-structure changes of white matter(WM)and the cognitive impairment thereof.Methods:The DW-MRI was performed in 30 probable AD patients and 30 normal controls with normal-appearing white matter(NAWM).The ADC was measured in different WM areas.The neurologic and neuropsychological assessments were examined with mini-mental state examination(MMSE)in patients.The ADC were determined in standard regions of the frontal,temporal,occipital and parietal white matter,genu,splenium of the corpus callosum.Results:The value of ADC was higher in frontal,splenium corpus callosum,temporal,and parietal white matter of AD group than that of control(P ＜ 0.01).There was no significant difference in the ADC value of genu and occipital white matter between AD and control groups(P＞ 0.05).The score of MMSE was 24.1±0.8 in AD group.The ADC values of parietal,splenium of the corpus callosum and frontal white matter were significantly negatively correlated with MMSE scores in AD group(P ＜ 0.05 or P ＜ 0.01).There was no correlation between the ADC values of genu of the corpus eallosum,temporal and occipital white matter with the MMSE score(P ＞ 0.05).Conclusion:The quantitative DWI analysis of MRI DWI may be helpful in assessing WM abnormalities in AD.The parietal WM abnormalities may play an important role in the development of dementia.It was showed that Alzheimer's cognitive decline with ADC value and micro-structure of white matter was closely related.

Aim To obtain two-dimensional gel electrophoresis maps of the prefrontal cortex(PFC)proteins of normal and morphine-addicted rats for identifying the diferentially expressed proteins in the addicted rats.Methods Rat models of morphine addiction were established.The proteins in the PFC underwent two-dimensiona1 gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertica1 SDS-PAGE as the second dimension.Analysis of 2-DE maps was used to determine differential expressions of proteins between the two groups by ImageMaster 2D Platinum v5.0,and four protein spots expressed differently were picked up for further identification by MALDI-TOF-MS.Results Matched and compared with those of control group,87 protein spots had been determined with differently expressive levels in morphine addiction group.By MALDI-TOF-MS,two protein spots of them had been identified as Snap25 Isoform ?-Snap 25 of Synaptosomal-associated protein 25 and Actb Actin,cytoplasmic 1.Conclusions There is obvious difference in expressive proteomes in PFC between normal and morphine-addicted rats. The functions of those identified proteins are involved in cell growth,apoptosis,differentiation,signal transduction,axon growth and cellular energy metabolism,so proteomics can serve as a new approach in the study of morphine dependence to discover new therapeutic targets.

The number average molecular weights of five fractionated samples of pomelo pectin were determined osmometrically in aqueous solution. The values of Mn ranged from 4.74?104 to 1.83?144 for different fractions.From the data of osmotic pressure and intrinsic viscosity, the [?]-M relation obtained is[?]3.23?10-7M1.75in 0.9% NaCl solution of pH 4.83 at 37℃.

Objective To construct the vector for ?-galactosidase (?-gal) gene expression regulation with tetracycline-regulated gene expression system,and to control ?-galactosidase gene expression in hepG2 cells with the vector.So that offer a experimental base for regulating gene expression for liver cancer gene therpy with this system.Methods The primers for explanding two tetracycline operator (TetO2) gene according to the gene nucleotide sequences of TetO2 gene and pcDNA3.1 vector were designed.TetO2 gene was amplified by PCR with pcDNA3.1 plasmid as template.The TetO2 PCR products were cloned into pcDNA3.1 and the vector was named as pcDNA3.1-TetO2.The pcDNA3.1-Teto2 with TetO2 PCR products sequence was analyzed by ABI3130 sequencing analysis.Then ?-gal gene was cloned into pcDNA3.1-TetO2,the vector was named as pcDNA3.1-TetO2-?-gal.The pcDNA6/TR plasmid vector with tetracycline repressor(TR) was transfected into HepG2 cells by Lipotap,and the stable transfection cells were screened by blasticidin.TR gene expression was detected by RT-PCR in HepG-2 cells with and without pcDNA6/TR plasmid transfection.The pcDNA3.1-TetO2-?-gal plasmid vector was transfected into HepG2 cells with and without pcDNA6/TR plasmid transfection,and 3 to 4 d later, these transfected gene cells were treated with doxycline(4 mg?L-1).After 48 h,?-gal gene expression was detected with ?-gal cell staining.Results The pcDNA3.1-TetO2 sequencing analysis results showed that a cassette was made for a cytomegalovirus-type 2 tetracycline operator (TetO2)-TetO2 promoter in pcDNA3.1-Teto2.The double digestion reslut of pcDNA3.1-TetO2-?-gal plasmid vector demonstrated that ?-gal gene was successfully cloned into pcDNA3.1-TetO2 vector .The RT-PCR result of TR gene showed TR gene could be expressed in HepG2 cells with pcDNA6/TR plasmid transfection and TR gene didn’t express in HepG2 cells without pcDNA6/TR plasmid transfection.?-gal gene expressed in HepG2 cells without pcDNA6/TR plasmid transfection,but didn’t express in HepG2 cells with pcDNA6/TR plasmid transfection.However,?-gal gene expression could be induced with doxycline in HepG2 cells with pcDNA6/TR plasmid transfection.Conclusion The tetracycline-regulated gene expression system vector for regulating ?-gal gene expression is successfully constructed and the vector system can control ?-gal gene expression in HepG2 cells.