OBJECTIVE:To study the protective effect of Weidean tablets on rats with stress-induced gastric ulceration. METH-ODS:Water immersion restraint stress method and empty stomach were employed to establish rat models with gastric ulceration. 60 SD rats were equally randomized into normal control group(isometric sodium chloride injection),model group(isometric sodium chloride injection),ranitidine group(0.015 g/kg),and groups of high,medium and low doses(1.70,0.87 and 0.43 g/kg)of Wei-dean tablets. General situation of stomach was observed .The gastric mucosal injury index,the NO content in serum,and the activi-ties of SOD and iNOS and the contents of MDA and PGE2 in the gastric tissue were detected for rats. Pathomorphological observa-tion of rats’gastric tissues was conducted under the microscope. RESULTS:Compared with normal control group,the rats in the model group had higher gastric mucosal injury index,lower content of NO in serum,and weaker activity of SOD,stronger activity of iNOS,higher content of MDA and PGE2,in the gastric tissue. There was statistical significance (P<0.01 or P<0.05). Dark matter,severe deformation and necrosis of gastric mucosal cells in rats were noted,as well as multiple large-area superficial ulcers. Compared with the model group,the rats in groups of high,medium and low doses of Weidean tablets had lower gastric mucosal injury index,higher content of NO in serum,and stronger activity of SOD,weaker activity of iNOS,lower content of MDA and higher content of PGE2,in the gastric tissue. There was statistical significance(P<0.01 or P<0.05). General situation of stomach and the pathomorphological condition of rats’gastric tissues were improved. CONCLUSIONS:Weidean tablets has protective ef-fect to some extent on rats with stress-induced gastric ulceration,the mechanism is related to the improvement of serum levels of anti-oxidation index in rats.

BACKGROUND: Limb allograft is a sort of composite tissues allotrans plantation(CTA), some researches showed that the apoptosis of target cell is one of the main mechanism of the dysfunction of allograft.OBJECTIVE: To investigate the characteristic of cell apoptosis in acute rejection of limb allograft in rats based on limb allograft model.DESIGN: A randomized controlled trial using the experimental animals as the objects.SETTING: Experimental animal center Laboratory of a hospital of a medical university MATERIALS: The experiment was done in the Experimental Animal Central Laboratory of the First Affiliated Hospital of Harbin Medical University from November 2003 to May 2004. Totally 56 healthy and male SD rats and 28 Wister rats were involved with body mass of 200 to 250 g. The rats were provided by the Experimental Animal Center of the First Affiliated Hospital of Harbin Medical University. They were randomly divided into two groups:transplantation group with 28 Wistar rats and 28 SD rats and control group with 28 SD rats.INTERVENTIONS: The transplantation group of SD rats underwent limb allotransplantation from allogenetic Wistar. The control group of SD rats underwent limb replantation. The expression of acute rejected in limb allografts was observed. The limb grafts were harvested atday 1, 3, 5 or 7 after transplantation. Histopathological rejection grade of each tissue rejection was performed with hematoxylin-eosin staining. Apoptotic cells were detected by TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) and apoptotic index(AI) was calculated.MAIN OUTCOME MEASURES: Primary results: ① istopathological grade of acute rejection in limb allografts ② The relationship between apoptosis and acute rejection in limb allograft in rats; Secondary results:General condition of rats in each group.RESULTS: The limb grafts showed edema and erythema and the skin became red at day(3.43 ±0.79) after transplantation. The average survival time was(7.42 ± 1.72) days. The acute rejection in skin was the strongest. On the day 3, 5 and 7 after operation, the histopathological rejection grades of skins in the transplantation group were(1.14±0.38) ,(2.28 ±0.48) and(2.86 ±0.38) grades respectively. They were significantly different from that of muscle and nerve( P ＜ 0.05 ) . The apoptotic cells in allografts were mainly infiltrating lymphocytes in subcutaneous tissues and then the muscle cells. All was positively correlated with acute rejection grade in limb allograft .CONCLUSION: Apoptosis was involved in acute rejection of limb allograft in rat. The apoptotic index can be used as a quantitative index to estimate the injury of grafts.

BACKGROUNDTo study to immunogenicity of recombinant HEV ORF2 protein expressed in pichia pastoris.

METHODSBALB/c mice were immunized with the recombinant HEV ORF2 protein. The ability of antiserum to bind HEV was tested using affinity-capture reverse transcription polymerase chain reaction (RT-PCR). Moreover, the recombinant protein was used to immunize BALB/c mice by different routes with different adjuvants. Serum conversion rate of anti-HEV antibody and the ELISA titer were detected.

RESULTSThe antiserum could capture native HEV for RT-PCR. As to the immunization effect, the immune response by intramuscular route was better than that of the intraperitoneal route. The protein with alum and CpG adjuvant could elicits more significant immune responses than using the alum adjuvant alone. The best way was to immunize with the protein with alum and CpG adjuvant by intramuscular route with a boosted injection on the 4th week after the first immunization. The ED50 was 0.023 microgram. This is the first report that the antibody elicited by recombinant HEV ORF2 protein expressed in pichia pastoris recognizes native HEV. High immunogenicity of this kind of ORF2 was also demonstrated by inducing strong immune response in mice with good ED50 result.

CONCLUSIONSThe high immunogenicity of this kind of HEV ORF2 may make a foundation for the development of new type of hepatitis E vaccine.

Animals ; Antibody Formation ; Female ; Hepatitis E virus ; immunology ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; Recombinant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

Diagnosis, Differential ; Humans ; Neoplasms, Glandular and Epithelial ; diagnosis ; Thymus Neoplasms ; diagnosis

OBJECTIVE:To explore the effect of Compound pueraria tablets on the kidney tissues of diabetic nephropathy rats. METHODS:High sugar and lipid diet combined with intraperitoneal injection of streptozotocin were used to establish the model of diabetic nephropathy (DN). Normal control group was established. Model rats were randomly divided into model control group, positive control group(Irbesartan tablet),Compound pueraria tablets low-dose,medium-dose and high-dose [0.102,0.203,0.406 g/(kg·d)] groups(n were 8-10). The corresponding drugs were given,and fasting blood glucose(FBG)and 24 h urinary protein (Upro) were collected at 1st,14th,28th,42nd,56th day after treatment. SCr,BUN,TC,TG and KI were detected,and renal pathology was observed after the last dose. RESULTS:Compared with normal group,FBG of those groups were all increased after modeling,and 24 h Upro of them were all increased after 28th day (P<0.05 or P<0.01). Compared with model control group, FBG of Compound pueraria tablets medium-dose and high-dose groups were all decreased since 42nd day (P<0.05 or P<0.01), and 24 h Upro of Compound pueraria tablets groups were all decreased since 28th day(P<0.05 or P<0.01);BUN,TC,TG and KI of Compound pueraria tablets in medium-dose and high-dose groups were all decreased significantly,and SCr of 3 dose groups were all increased (P<0.05 or P<0.01). The morphological structure of renal cells was improved significantly in drug treatment groups. CONCLUSIONS:Compound pueraria tablets can correct lipid metabolism disorder,reduce Upro and improve renal func-tion,indicating certain protective effect on the kidney tissues of diabetic nephropathy rats.

OBJECTIVE:To study the effect of Compound pueraria tablets on osteoporosis model rats induced by retinoic acid. METHODS:The osteoporosis model was induced by intragastric administration of retinoic acid solution for 15 days;normal group was established. After modeling,the rats were randomly divided into model control group,Xianling gubao capsule [0.32 g/(kg·d)] positive control group,Compound pueraria tablets low-dose and high-dose [0.24,0.4 g/(kg·d)] groups (n=8). After 6 weeks of ig,the serum sample was collected to determine the levels of serum calcium(s-Ca),serum phosphorus(s-P),ALP and bone gla protein (BGP);bone density instrument was used to detect the contents of bone mineral density (BMD),bone mineral content (BMC),bone image area (BIA) and muscle content (MC);the results of compact bone substance scanning were observed. RE-SULTS:Compared with normal group,the levels of s-Ca,BMD,BMC and MC in rats were decreased in model control group, while the level of BGP was increased(P<0.05 or P<0.01). Compared with model control group,related index and compact bone substance scanning of Compound pueraria tablets groups were all improved;the levels of s-Ca,s-P,ALP,BMD and MC were in-creased in Compound pueraria tablets high-dose group,while the level of BGP was decreased;the levels of BMD and MC were in-creased significantly in Compound pueraria tablets low-dose group(P<0.05 or P<0.01). CONCLUSIONS:Compound pueraria tab-lets can improve the osteoporosis induced by retinoic acid in rats.

Objective To investigate the clinical application and manifestation of dynamic contrastenhanced MRI (DCE-MRI) in differentiating true progession from pseudoprogression in patients with gliobastomas.Methods Twenty five glioma patients were treated with postoperative concurrent chemoradiotherapy and enrolled in this study.All patients were underwent DCE-MRI using a 1.5T scanner.Fifteen patients were confimmed by secondary pathology or clinical and imaging follow-up of patients with gliomas true progession (TP),10 patients were pseudoprogress (PP).Nonparametric Mann-Whitney test was used to compare perfusion parameters between two groups (TP and PP),were used for receiver operating characteristic (ROC) curve analysis to clear if these parameters can be the indicators to differentiate true progession from pseudoprogression.Results Ktrans (volume transfer constant),Ve (fractional volume of extravascular extracellular) values between TP and PP glioma groups were statistically significant,K and Ve values were significantly higher in the TP group than in the PP group (P ＜ 0.05).The areas under the ROC curve are 0.990 and 0.847,respectively.Kep (efflux rate constant) value,Vp (fractional volume of plasma) value in the identification of glioma TP group and PP group was not statistically significant (P ＞ 0.05).Conclusions DCE-MRI can be used to identify glioma TP and PP,Ktrans value and Ve value have clinical significance.

A simple,rapid and sensitive liquid chromatography-mass spectrometry（LC-MS）method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic ether-isopropanol（2∶1）and the extraction was performed on a Kromasil C18 column（150 mm×4.6 mm,5 μm）with the mobile phase of methanol-water（41∶59,v/v）within a run time of 6.0 min.The analyte was monitored with positive electrospray ionization（ESI）by selected ion monitoring（SIM）mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard（IS）geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.

BACKGROUNDTo determine the antigenicity of recombinant hepatitis E virus ORF2 (rHEV ORF2) protein expressed in Pichia pastoris (P. pastoris).

METHODSBy using the rHEV ORF2 protein from E.coli as control, an indirect ELISA was adopted to identify the sensitivity, specificity and stability of rHEV ORF2 protein from P. pastoris in detection of HEV IgM and IgG antibody in sera from patients with hepatitis E. The reactivity of the rHEV ORF2 against 5 HEV ORF2 monoclonal antibodies (McAbs) was also tested.

RESULTSThe minimum concentration of coated antigen with which HEV IgG could be detected was 12.5 ng/ml, while the highest serum dilution to detect both IgM and IgG antibodies against HEV was 1:5 120. No cross-reaction was found with sera from patients with any other types of hepatitis. The 37 degree C acceleration test showed that the rORF2 was highly stable within 12 months at 4 degrees C. The 5 HEV ORF2 McAbs showed better reaction with the rORF2 from P. pastoris, especially that 4B2, 2E2, whose reaction against the rORF2 were 125 and 25 times respectively higher than that of rORF2 from E.Coli.

CONCLUSIONThere may be more extensive conformational epitopes in the rHEV ORF2 from P. pastoris. The excellent antigenicity, sensitivity and stability suggest that it can be served as a new candidate antigen for the development of diagnostic reagents of hepatitis E.

Gene Expression ; Hepatitis Antibodies ; blood ; Hepatitis Antigens ; genetics ; immunology ; Hepatitis E ; immunology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

OBJECTIVETo investigate the effects of the new antiallergic agent N-(pyridin-4-yl)-(indol-3-yl) acetamide-45 (acetamide-45) on electric field stimulation (EFS)-and methacholine-induced contraction of airway smooth muscle in vitro.

METHODSContractions were induced by EFS in isolated trachea and bronchus of rats or by cumulative methacholine concentrations in isolated trachea of guinea pigs. Changes in isometric force of isolated airway smooth muscle were measured by force transducers and recorded on a multi-channel polygraph recorder.

RESULTSAcetamide-45 inhibited the contraction induced by EFS in isolated rat airway. The IC50 was 10.74 (95% CI 8.87-13.00) micromol.L(-1) and 18.83 (95% CI 14.57-24.33) micromol.L(-1) in tracheae and bronchi, respectively. Acetamide-45 also inhibited methacholine-induced contractile response of isolated guinea pig trachea in a concentration-dependent manner. At concentrations of 3, 10, 30 micromol.L(-1) acetamide-45 significantly decreased maximal contractile response of methacholine by 24.6%-43.2% and increased EC50 of methacholine by 3.1-to 21.4-fold.

CONCLUSIONAcetamide-45 inhibits EFS-or methacholine-induced contraction of isolated airway smooth muscle, and these effects might be non-specific inhibition on cholinergic receptor.

Acetamides ; pharmacology ; Animals ; Depression, Chemical ; Electric Stimulation ; Guinea Pigs ; In Vitro Techniques ; Male ; Methacholine Chloride ; pharmacology ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Trachea ; drug effects