italic>Cannabis sativa, one of the ancient medicinal plants, has been used to alleviate pain and seizures. However, cannabinoids are often addictive, which limits their clinical use. Cannabidiol (CBD) as a non-psychoactive component of Cannabis sativa, has much weaker adverse effects than Δ9-tetrahydrocannabinol (THC) and therefore has received widespread attention. CBD has been found to ameliorate a variety of neuropsychiatric diseases, but the precise mechanism(s) of action are still unclear. Due to its low affinity for classical cannabinoid receptors current studies are focusing on other targets outside the endocannabinoid system. In the present review we mainly summarize the effects and molecular mechanisms of CBD in neuropsychiatric disorders, including epilepsy, neuropathic pain, anxiety, and depression.

OBJECTIVE:To discuss the approach to launch pharmaceutical care in hospital preparation room.METHODS:The contents of launching pharmaceutical care in the preparation room of our hospital were analyzed and summarized retrospectively.RESULTS & CONCLUSION:A shifting toward technical service is on the way by launching pharmaceutical care in hospital preparation room.

Objective To screen the proteins binding with HBV X-promoter and to investigate their potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV X biotinylated promoter DNA as the selective molecule, the T7 select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment and cloned into pGEM-Teasy vector. Thirty positive plaques were chosen for DNA sequencing. Results Five kinds of known and nine kinds of novel cDNA sequences were obtained. The 5 kinds of known ones are human serum albumin, NADH dehydrogenase subunit 4, prokininase, ubiquitin specific protease 10, and testis enhanced gene transcript. Conclusion The binding proteins of HBV Xp DNA were identified by phage display. The results suggest that this approach provides a new search tool for the study of replication and expression mechanism of HBV DNA.

Objective To explore the correlation between intestinal mucosal immune barrier and pathogenesis of pigment gallstone and its possible mechanism.Methods Eighty guinea pigs were randomly divided into control group(group CON),pigment gallstone group(group PS),and intestinal mucosal protection group(group GLN).The guinea pigs were fed with normal diet in group CON,pigment gallstonein during diet in group PS,and glutamine-supplemented diet in group GLN for 8 weeks.The guinea pig model of pigment gallstone was established.The incidence of pigment gallstone was detected.The morphology of intestinal mucosa was observed,and the numbers of CD3~+T cell,CD40~+B cell,and IgA~+ plasma cell were counted.Results The incidence of pigment gallstone was significantly higher in group PS than in groups GLN and CON(P＜0.05).Compared with group CON,the intestinal wall was significant thinner and represented obvious signs of inflammation in group PS,and the numbers of CD3~+ T cell,CD40~+ B cell,and IgA~+ plasma cell significantly decreased(CD3~+ T cell,21.8±2.5 vs 11.1±3.4,P＜0.01;CD 40~+B cell,12.9±2.0 vs 10.7±3.6,P＜0.01;IgA~+ plasma cell,12.4±3.4 vs 10.7±3.5,P＜0.01).The signs of inflammation were less severe in group GLN than in group PS.There were significant differences in the numbers of CD3~+ T cell,CD40~+ B cell,and IgA~+plasma cell between groups GLN and PS.Conclusion Intestinal barrier dysfunction,including mechanical barrier and immune barrier,is involved in the formation of pigment gallstone.Glutamine has proved to improve the function of intestinal mucosal barrier and decrease the incidence of pigment gallstone.

Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

Objective To analyze the levels and speciation of arsenic metabolites in urine of rats treated with sodium arsenite and sodium arsenate in order to investigate the different aspects of metabolism between sodium arsenite and sodium arsenate,thus to understand further the basic data about relationship between it's metabolism and mechanism of toxicity. Methods Seventy Wistar rats,weighting 80-120 g,were divided into 7 groups of 10 each,such as normal control group,high,middle and low sodium arsenite group and high,middle and low sodium arsenate group. After the animals were fed for one month,the urine was collected by metabolic cage in 12 hours. Applying the high efficiency liquid chromatography and hydride genesis atomic fluorescence spectroscopy (HPLC-HGAFS),the levels and speciation of arsenic metabolites were determined in urine of rats. Meanwhile,the recovery rate of dimethyl arsinic acid(DMA) would be determined to estimate the degree of accuracy of results. Results The levels of iAs~(3+),iAs~(5+) and DMA in middle sodium arsenite group[(121.66±1.26),(10.26±2.68),(200.91±0.56) μg/L]were higher than the high sodium arsenite group[(113.20±0.75),(5.16±1.32),(147.70±μ0.77)μg/L,all P < 0.05]and low sodium arsenite group[(79.35±2.12),(5.13±2.25),(56.35±1.23)μg/L,all P < 0.05]. The levels of iAs~(3+) and DMA in middle sodium arsenate group[(315.81±1.69),(245.12±1.18)μg/L]were higher than the high sodium arsenate group[(85.03±0.56),(110.34±1.04)μg/L,all P< 0.05]and low sodium arsenate group[(22.97±2.67),(15.75±2.15)μg/L,all P < 0.05]. Compared with sodium arsenate group,the levels of iAs~(3+) and DMA in high and low sodium arsenite group were higher(all P < 0.05) ; and the levels of iAs~(3+) and DMA in middle sodium arsenite group were lower(all P < 0.05). Meanwhile,the average urinary recovery rate of DMA of rats in different sodium arsenite group were 94.80%-102.70%,and the average urinary recovery rate of DMA of rats in different sodium arsenate group were 95.33%-108.40%. Conclusion The speciation and levels of arsenic are influenced by the external exposure dose,and some distinction appeared in the metabolism and metabolic path between sodium arsenite and sodium arsenate in urine in vivo.

Objective To investigate the effects of the silence of teratocarcinoma-derived growth factor-1 ( TDGF-1 ) gene on invasion of human pancreatic cancer cell. Methods Three small interfering RNA (siRNA) targeting for TDGF-1 genes (S1, S2, S3 ) were designed and established, then the gene with the best silencing effects was screened. Human pancreatic cancer cell line PANC1 were transfected by siRNA with different concentrations (3. 125, 6.25, 12.5 nmoL/L), the cells without transfection, and simply treated with liposomes were controls. The expressions of mRNA and protein of TDGF-1 were determined by real time PCR and Western blot assay, respectively. The anchorage-independent growth was examined by clon formation in soft agar, and invasion ability was evaluated by boyden chamber model. PANC1 cells with transfection for 48h were injected into the nude mice to evaluate the invasion ability in vivo. Results The expressions of TDGF-1 mRNA and protein of cells transfected by siRNA were decreased in a dose-and time-dependent manner, which were significantly lower than those in liposomes group. Number of colony formation and transmembrane cell were 19.8 ± 2.2 and 49.8 + 2.6 in the control group, and 5.6 + 1.2 and 8. 1 + 1.1 in the 12.5 nmol/L transfection group. The volumes of tumor 4 weeks after transplation in the control group, liposomes group and the 12.5 nmol/L transfection group were (2.228 ± 0.016 ) cm3, ( 2.186 ± 0.028 )cm3 and ( 0.728 ± 0.023 )cm3. Conclusions TDGF-1 gene silence could inhibit invasion ability of human pancreatic cancer cell PANC1.

AIM:To investigate the effect of berberine on IL-1 or tumour necrosis factor (TNF) induced polymorphonuclear leucocyte(PMN)-endothelium adhesion and adhesion molecules.METHODS:Based on the model of human umbilical vein endothelial cell (HUVEC), this study adopted Rose Bengal Stain, cell ELISA, immunocyto-chemical techniques to investigate the effect of berberine on PMN-endothelium adhesion and the expression of cell adhesion molecules (CAMs).RESULTS:Berberine inhibited IL-1, TNF-induced HUVEC adhesion for PMN when pretreated HUVEC and antagonised IL-1, TNF-induced upregulation of ICAM-1 on HUVEC. Meanwhile, TNF-stimulated PMN adhesion for HUVEC and CD18 upexpression on PMN was diminished in the presence of berberine.CONCLUSION: Inhibite PMN-endothelium adhesion by downregulating the CAMs expression to inhibite PMN migration across endothelium is one of the mechanisms of antiinflammation of berberine.

Objective:To study the cytokine mechanism of Jiawei Xuanfei Toujie Ji(JXT)therapy to influenza infected mouse.Methods:Mice were inoculated intranasally with mouse lung-adapted influenza virus strain(FM1),and were treated with JXT. At 5th days after postinoculation were sacrificed, and IL-2,TNF-?,IL-6,IFN-? in their sera were measured with ELISA kits, their lungs sections were stained with hematoxylin and eosin.Results:Compared with model group ,the levels of IL-2 and IFN-? in JXT group were dramatically higher,and the levels of TNF-? and IL-6 were lower,and pathological effects of influenza pneumonia showed less lung injury.Conclusion:Regulative effects on cytokine may be one of the factors that JXT decrease the pathogenesis of influenza-induced acute lung injury.

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.