OBJECTIVE:To establish a method for determining the contents of chlorogenic acid and luteoloside in Lonicerae ja-ponicae,and to explore the shelf life of L. japonicae under ordinary temperature and sealed environment. METHODS:HPLC meth-od was adopted. The determination was performed on Agilent Zorbax SB-C18(chlorogenic acid)column and Agilent Zorbax SB-Phe-nyl(galuteolin)with mobile phase consisted of acetonitrile-0.4% phosphoric acid(13:87,V/V),acetonitrile-0.5% glacial acetic ac-id (gradient elution,galuteolin) at the flow rate of 1.0 mL/min. The detection wavelength was set at 327 nm (chlorogenic acid) and 350 nm(galuteolin). The column temperature was 30 ℃,and sample size was 10 μL. RESULTS:The linear range of chloro-genic acid and galuteolin were 10-100 μg/mL(r=0.9986),5-50 μg/mL(r=0.9993),respectively. RSDs of precision,stability and reproducibility tests were all lower than 4.0%. Recoveries were 95.78%-99.70%(RSD=1.46%,n=6)、96.30%-104.31%(RSD=2.93%,n=6). The contents of chlorogenic acid and galuteolin in roller method,baking method and natural drying method of processed L. japonicae were all decreased by 30%-40% after stored for 12 months. CONCLUSIONS:The method is simple, precise,stable and repeatable,and can be used for simultanoue determination of chlorogenic acid and luteoloside in L. japonicae. The contents of active components in L. japonicae decrease significantly after stored for 12 months. It is necessary to establish shelf life standard of L. japonicae,so as to guarantee the effectiveness of drug use in the clinic.

Objective To investigate 25S rDNA genotype distribution of vaginal Candida albicans and its relationship with antifungal susceptibility. Methods A total of 954 isolates of Candida albicans were subjected to genotype analysis according to the presence and the size of transposable groupⅠintron in 25S rDNA with PCR; 156 isolates were enrolled to detect their antifungal susceptibility of 5 antifungal agents with the M27-A3 broth microdilution method. Results Totally 876 isolates were determined to be intronless genotype A (91.8%,876/954); 58 (6.1%, 58/954) and 20 isolates (2.1%, 20/954) were intron-containing genotype B and genotype C, respectively. The minimal inhibitory concentrations (MIC) of itraconazole and fluconazole against the intron-containing group were significantly higher than those of the intronless group (0.25 versus 0.125μg/ml, P<0.05;0.25 versus 0.125μg/ml, P<0.01), whereas the MIC of nystatin against the intron-containing group was lower than that of the intronless group (4 versus 8μg/ml, P<0.01). The resistance rate of Candida albicans to itraconazole of the intron-containing group was significantly higher than that of the intronless group [24% (19/78) versus 3% (2/78), P<0.01]. Cross-resistance analysis revealed 5 of the 21 (24%) itraconazole resistant isolates were cross-resistant to fluconazole ,and 5 of the 6 (5/6) fluconazole-resistant strains were cross-resistant to itraconazole. Besides, all the five strains simultaneously resistant to itraconazole and fluconazole belonged to the intron-containing group. Conclusion The presence of the transposable group Ⅰ intron in 25S rDNA of vaginal Candida albicans might be important in affecting itraconazole, fluconazole and nystatin susceptibility.

Objective To investigate apoptosis in acinar cells during posttransplant reperfusion, and the expression of Bcl 2, Bax, Fas Methods Pancreas transplantation was performed between male SD rats after 2 hours of cold ischemia Rats ( n =65) were divided at random into 7 groups: sham group, cold ischemia group (as controls), reperfused 1 h, 3 h, 6 h, 9 h, 12 h group The pathologic pancreatic tissue was observed by electron microscope; Apoptotic cells were stained by TUNEL technique; Expression of Bax、Bcl 2 and Fas protein in pancreatic tissue were detected by SABC immunohistochemical method Results Apoptotic cells were observed in early phase after pancreas transplantation The peak of apoptotic pancreas cells occurred 3 hours after reperfusion [AI ( 9 5? 2 9)%, P

Objective:To establish the model of pancreaticoduodenal transplantation in rats. Methods:The enteric drained whole pancreas duodenal transplantation model was investisated. The donor aortic segment including the celiac, superior mesenteric divergence were anastomosed with recipient abdominal aorta with end to side. The donor portal was cuff-anastomosed with left renal vein. The graft duodenum were anastomosed with the host jejunum with end to side. Results:Eighty-nine operations were performed and 51 rats were successful, of which all graft of successful case had endocrinium function. Conclusion:This model for pancreas transplantation is reliable.

Lysophosphatidic acid is a intermediary metabolite in lipid metabolism, mainly produced from activated platelets. It may cause the apoptosis of hippocampal neurons, abnormal phosphorylation of tau protein and increased permeability of the tight junction of endothelial cells, which is closely correlated with pathological changes of ischemic cerebrovascular diseases, Alzheimer's disease and vascular dementia. It is shown that lysophosphatidic acid may induce the opening of the blood-brain barrier under pathological conditions, and enters central nervous system through the blood-brain barrier, and causes different effects of pathological changes by acting on various cells.

Objective To explore the roles of glycogen synthase kinase-3β (GSK-3β) in cognitive dysfunction and emotional alterations after rat's chronic cerebral ischemia.Methods 51 SD rats were randomly divided into sham group (sham group,n=17),chronic cerebral ischemia group (2VO group,n=17),chronic cerebral ischemia group + LiCl group (2VO + LiCl group,n=17),according to the table of random number.All groups were intraperitoneally injected with LiCl or saline on 3rd,7th,14th,21 st and 28th day,and then produced the chronic cerebral ischemia models.On the 28th day after model,the spatial learning and memory,fear memory,and anxiety emotion were detected.Results The Morris water maze test showed that 2VO group spent longer latent time searching and finding the platform than sham group (4th day P＜0.01,3rd,5th,6th,7th day P＜0.05).2VO+LiCl group spent shorter latent time than 2VO group (4th day P＜0.01,5th,6th,7th day P＜0.05).After removing the platform,2VO group spent longer time arriving the former location than sham group (P＜0.05).And 2VO+LiCl group spent dramatically different time compared to 2VO group (P＜0.01).Step-down test showed 2VO group spent shorter latent time than sham group (2VO group:(41.00±1.87)s,sham group:(44.55±2.77)s) (P＜0.05).2VO+ LiCl group spent dramatically longer latent time compared to 2VO group (2VO +LiCl group:(43.40± 1.35)s) (P＜ 0.05).2VO group made much more mistakes times than sham group (P＜0.05).2VO+LiCl group made dramatically less mistakes times compared to 2VO group (P＜0.05).The elevated plus maze test showed 2VO group had much less ratio of retention time in open arms (among total arms retention time) than sham group (2VO group:(0.23± 0.01),sham group:(0.25± 0.01)) (P＜ 0.01).2VO + LiCl group had much larger ratio than 2VO group (2VO + LiCl group:(0.24±0.01),P＜0.05).2VO group had much less ratio of entry times in open arms (among total arms entry times) than sham group (P＜0.01).2VO+LiC1 group had much larger ratio than 2VO group(P＜0.05).Conclusion Chronic cerebral ischemia can lead to deterioration of learning and memory,and anxiety emotion for rats.However,inhibition of GSK-3β can ameliorate these alterations.

Objective The aim of this study was to investigate the relationship between juxtapapillary duodenal diverticula (JDDs)and stenosing papillo odditis,and the diagnosis and treatment for this clinical entity.Methods Patients underwent upper gastrointestinal radiography,ERCP, and radionuclide scan. 18 cases were treated with operation,8 cases underwent sphincteroplasty,3 with combined diverticulo sphincteroplasty, 3 with choledochoduodenostomy or choledochojejunostomy,1 with gastrojejunostomy,1 case with diverticulectomy.Results Sphincteroplasty effects a good result for coexistant JDDs and stenosing papilloodditis. There were no recurrence and complication after the combined diverticulo sphincteroplasty. Conclusion There was a causal relation between JDDs and stenosing papillo odditis. Effective treatment was achieved by combining sphincteroplasty with diverticulo sphincteroplasty.

AimTo investigate the influence of tetramethylpyrazine(ligustrazine, Lig) on oxygen consumption and superoxide during the respiratory burst of human neutrophils. MethodsIt was observed by using ESR spin trapping, spin probe oxymetry and luminol-dependent chemiluminesence(CL) . Results Lig had no influence on oxygen consumption during the respiratory burst of neutrophils(P＞0.05), but had remarkable inhibition on CL response generated by neutrophils(P＜0.01), and had scavenging effect on O2 and OH ·generated by neutrophils, which were demonstrated in xanthine/ xanthine oxidase system and Fentons reaction(P＜0.01) . ConclusionLig has no inhibiting effect on oxygen metabolic function of neutrophils,but protects tissue from injury caused by activated neutrophils through scavenging oxygen radicals.

OBJECTIVE: To investigate the effect of high glucose on the expression of c-met in human kidney fibroblasts in vitro, and to explore the regulation of Radix Astragali. METHODS: A cell culture system of human kidney fibroblasts was developed in vitro. The human kidney fibroblasts were divided into normal control group, high glucose group and mannitol group. Expressions of c-met and transforming growth factor-beta1 (TGF-beta1) mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR) and the expressions of c-met protein were analyzed by Western blot method after 6-, 12-, 24-, 48- and 96-hour culture. The human kidney fibroblasts were also cultured with 10% Radix Astragali containing serum; the expressions of c-met mRNA and protein were detected after 24- and 48-hour culture. RESULTS: Compared with the normal control group, expression of c-met mRNA in the high glucose group was significantly increased after 12-hour culture (P<0.05), arriving at the peak after 24-hour culture (P<0.01). The level of TGF-beta1 mRNA was higher in the high glucose group than that in the normal control group after 24-hour culture (P<0.05), arriving at the peak after 96-hour culture (P<0.01). Forty-eight hours after treating with 10% Radix Astragali containing serum, the levels of c-met mRNA and protein in fibroblasts were increased, and were higher than those in the high glucose group (P<0.01, P<0.05). CONCLUSION: High glucose can induce the expressions of c-met mRNA and protein in earlier period, and then inhibit the expressions. Radix Astragali can up-regulate the expressions of c-met mRNA and protein of human kidney fibroblasts, which may be one of its action mechanisms in delaying the progression of diabetic nephropathy.

Objective To investigate the effect of repetitive transcranial stimulation (rTMS) on learning and memory and on the neuron and synapse ultrastructures of the CA3 region of the hippocampus.Methods Forty Sprague-Dawley (SD) rats were randomly divided into 4 groups (n =l0 in each group):a normal control group,a depression group,an rTMS group and a sham group.Unpredictable mild stress was used to establish depression models in the rats of the latter3 groups.The sucrose water consumption test and open-field test were used to evaluate any depressive behavior of each group.The rTMS group rats were given 15 Hz rTMS for 21 days while the sham group received sham stimulation.The orientational navigation and spatial probe tests were performed on each group using a Morris water maze to evaluate their learning and memory abilities.In addition,changes in the ultrastructure of the CA3 region of the hippocampus were detected using transmission electron microscopy.Results The modelling induced significant differences in the sucrose water consumption test results and in horizontal and vertical behavior in the open-field tests.Escape latency and spatial probe time were significantly different between the rTMS group and the sham and depression groups.There was no significant difference in the behavioral indexes between the depression group and the sham group.Electron microscopy showed pathological changes in the ultrastructures of the neurons and synapses in the CA3 region of the hippocampus among the depression group,while in the rTMS group those ultrastructures tended to be basically normal.Conclusion rTMS can improve learning and memory during depression,at least in rats.A possible mechanism is that rTMS can induce changes in the ultrastructures of neurons and synapses in the CA3 region of the hippocampus.