Objective To establish a HPLC method for simultaneous determination of uracil,hypoxanthine,guanosine, thymine,and( R,S)-goitrin in Isatidis Radix. Methods The determination was performed on a Hanbon Hedera C18 column (4. 6 mm×250 mm,5 μm). The mobile phase consisted of acetonitrile( A)and water( B)with linear gradient elution at the flow rate of 1.0 mL·min-1. The column temperature was 30 ℃. Detection wavelength was 254 nm. Results The linear range of uracil,hypoxanthine,guanosine,thymine,and(R,S)-goitrin was 1.97-39.40 mg·L-1(r=0.999 9),1.25-50.00 mg·L-1(r=0.999 9),0.25-10.40 mg·L-1(r=0. 999 6),2. 84-56. 70 mg·L-1(r=0. 999 9),and 0. 72-28.80 mg·L-1(r=0.999 8), respectively. The average recovery was 99. 36%(RSD=0. 91%),99. 79%(RSD=1. 12%),100. 90%(RSD=1. 71%),98. 67%(RSD=1. 50%),and 99. 33%(RSD=0. 94%),respectively. Conclusion The method is simple,accurate,reliable,reproducible and sensitive,which can be used as an effective method for quality control of Isatidis Radix.

Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20％ of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.

Objective To investigate the effect of propofol anesthesia on the expression of β-secretase 1 (BACE1) and content of anyloid beta protein 1-42 (Aβ1-42) in the neonatal rat hippocampus.Methods Ninety Sprague-Dawley rats,aged 7 days,weighing 12-16 g,were randomly divided into 3 groups ( n =30 each):control group (group C),single dose of propofol anesthesia group (group SP),and repeated doses of propofol anesthesia group (group RP).Group C received intraperitoneal normal saline 7.5 ml/kg once a day for 7 consecutive days.Group SP received normal saline 7.5 ml/kg once a day for 6 consecutive days and propofol 75 mg/kg on 7th day.Group RP received propofol 75 mg/kg once a day for 7 consecutive days.Six rats in each group were chosen at 15 min after the end of injection on 7th day and blood samples were taken from the left ventricle for determination of the blood glucose level and for blood gas analysis.Eight animals in each group were sacrificed on 1st,3rd and 7th day after the end of injection on 7th day to determine the expression of BACE1 (using Western blot) and content of Aβ1-42 in the hippocampus (by ELISA).Results Compared with groups C and SP,the expression of BACE1 was up-regulated and the content of Aβ1-42 was significantly increased at each time point in group RP ( P ＜ 0.01 ).There was no significant difference in the expression of BACE1 and content of Aβ1-42 at each time point between groups C and SP ( P ＞ 0.05).Conclusion Repeated doses of propofol up-regulate the expression of BACE1 and increase the content of Aβ1-42 in neonatal rat hippocampus,which may be one of the mechanisms by which propofol leads to long-term cognitive dysfunction.Single dose of propofol does not have the effect.

ObjectiveTo investigate the effect of propofol repeated anesthesia on the expression of CaMK Ⅱ α in the hippocampus in neonatal rats.MethodsThirty-two SD rats aged 7 days weighing 12-16 g were randomly divided into 2 groups (n =16 each): group C received intraperitoneal 0.9％ normal saline 7.5 ml/kg once a day for 7 days and group P received propofol 75 mg/kg once a day for 7 days.Learning and memory function were assessed using Morris warier maze at 28 days old of rats.The animals were sacrificed at 24 h after the tests and brain tissues were removed.The expression of CaMK Ⅱ α and phosphorylated CaMK Ⅱ α (pCaMK Ⅱ α) in hippocampal CAI region were determined by immunochemistry and Western bolt.ResultsCompared with group C,the escape latency was significantly prolonged,space exploration time shortened and expression of CaMK Ⅱ α and pCaMK Ⅱ α down-rugulated in group P than in group C( P ＜ 0.01 ).ConclusionPropofol repeated anesthesia decreases congnitive function through down-regulating the expression and inhibiting the activity of CaMK Ⅱ α in hippocampus in neonatal rats.

Objective HupA is one of the potential drugs which can be used to treat Alzheimer's disease(AD).The aim of this study was to explore the pharmacokinetics and biodistribution of HupA in vivo by using 11C-HupA.Methods A total of 25 SD rats were studied.They were divided into 5 groups (5 rats in each group).All had intravenous injection of 22 MBq(in0.2 ml)11C-HupA through tail vein.Dynamic im-aging Was acquired from 5 to 90 minutes after injection.Venous blood and organ activities were collected at 5,15,30,60.and 90 minutes after injection.Percentage activity of injected dose per gram of tissue(%ID/g)was calculated to characterize the biodistribution of tracer in different brain regions: frontal,apical, temporal,occipital,cerebellum,hippocampus,striatum,thalamencephalon, and brain stem, Variance analysis using SPSS 11.5 software was performed and compared among the study groups.Results 11C-HupA was character-istic for its quick clearance from blood,with half time T1/2 of (14.61±1.77) min,and clearance rate (CL)macokinetics of 11C-HupA in rats corresponded to a one-compartment model.with an activity curve(area 11C-HupA distribution in different brain regions,being greater in cerebral cortex,hippocampus,hypothala-mus and brain stem. Conclusions Pharmacokinetic study of 11C-HupA in brain was fast.convenient and showed high specificity and sensitivity.Its ability to quantitatively evaluate brain function and its character-istic distribution in mice provided some evidence for monitoring therapy in AD patients.

Objective To study the efficacy and mechanism of compound Tripterygium hypoglaucum Hutch (THH) on photoallergic contact dermatitis in mice. Methods The photoallergic animal model of BALB/c mice was established by using photosensitizer chlorpromazine and UVA irradiation. The therepeutic efficacy was determined by measuring the thickness and the weight of the swelling ear and the number of infiltrated mononuclear cells in the ear tissue. Immunohistochemical technique was used to detect the ICAM-1 expression on keratinocytes, fibroblasts and vascular endothelial cells. The serum level of INF-? was measured by ELISA. The tested animals were divided into 3 groups: compound THH, THH alone and normal saline. Results The difference of the thickness of left ear before and after challenge, the differences of the thickness and the weight of ear tissue, the difference of the number of infiltrated mononuclear cells of left and right ear after challenge were significantly less in the compound THH group than those in the THH alone group (P

OBJECTIVETo observe the effects of Xiaobai Decoction (XBD) in reducing albuminuria and shortening the duration of albuminuria in patients with pregnancy-induced hypertension syndrome (PIH) in puerperium.

METHODSEighty-five patients were given the conventional treatment with magnesium sulfate for relieving convulsion and lowering hypertension, at the same time, the treated group was given XBD additionally with the modification according to the symptoms. The treatment course for both groups was 14 days. Routine test of midstream urine was performed every three days, and 24 h-urinary protein was measured every week.

RESULTSThe therapeutic effect on the 43 patients of the treated group was markedly effective in 11 (25.6 % ), effective in 26 (60.4%) and ineffective in 6 cases (14.0%), the total effective rate being 86.0%; while in the 42 patients of the control group, the corresponding numbers were 5 (11.9%), 21 (50.0%), 16 (38.1%) and 61.9%, respectively, the efficacy of the former was significantly better (P < 0.05).

CONCLUSIONXBD is a simple, safe and effective drug for reducing albuminuria and shortening the duration of albuminuria in puerperium of PIH patients.

Adult ; Albuminuria ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension, Pregnancy-Induced ; drug therapy ; Phytotherapy ; Postpartum Period ; Pregnancy ; Treatment Outcome ; Young Adult

Objective To explore the characteristics of proinsulin secretion in latent autoimmune diabetes in adults (LADA). Methods Fasting and 2 h sera in oral glucose tolerance test from 36 LADA patients, 37 type 2 diabetic patients and 43 healthy controls were collected to test glucose, proinsulin (PI) and C-peptide (CP) by radioimmune assay. Glutamie acid decarboxylase antibodies (GAD-Ab) were determined by radioligand assay.Results (1) Fasting proinsulin (FPI) and 2 h proinsulin (PPI) level in LADA patients were lower than those in type 2 diabetic patients (P<0.05), being both significantly inereasad compared with healthy controls (P<0.05 or P<0.01); The ratios of FPI/FCP and PPI/PCP (%) in LADA were beth significantly higer than those of type 2 diabetes mellitus and controls (P<0.05 or P<0.01); (2) LADA type-1 (GAD-Abe>0.3) patients showed lower PI levels(P<0.05 or P<0.01) and higher PI/CP ratio (all P<0.05) than LADA type-2 (0.05≤GAD-Ab<0.3); Meanwhile, there were no significant differences in above parameters between LADA type-2 and type 2 diabetes meUitus (P>0.05). (3) GAD-Ab index was negatively correlated with FPI and PPI in LADA group (r=-0.236 and-0.268, both P<0.05), and positively correlated with PPI/PCP (r=0.254, P=0.030).Meanwhile BMI was positively correlated with FPI, PPI and PI/CP in type 2 diabetes mellitus (all P<0.01). No factor entered the multiple regression analysis for predieting the hyperproinsulinemia and dispropriately elevated proinsulin levels in LADA patients. (4) According to the 99.5 th percentile of proinsulinemia in the healthy controls, which is defined as the cutoff point dispropriately elevated proinsulin levels, the proportion of subjects with fasting dispropriately elevated proinsulin levels (FPI/FCP) were 77.8%, 62.2% and 2.3% in LADA, type 2 diabetes meUitus and controls respectively, and PPI/PCP 83.3%, 51.4% and 2.3% respectively. Conclusion LADA patients, as well as type 2 diabetic patients, all showed hyperproinsulinemia and disproportionately elevated proiasulin levels that were one of characteristics of defective β-cell function. Moreover, disproportionately elevated nproinsulin level is more evident in LADA patients than that in type 2 diabetics and this may be related to humoral immunity.

Objective To investigate the effects of different modes of one-lung ventilation (OLV) on hemodynamies in the patients undergoing thoracic operation.Methods Forty-five adult patients undergoing thoracic surgery,were randomly allocated into 3 groups based on the modes of OLV used ( n =15 each):intermittent positive pressure ventilation ( IPPV,VT 6-8 ml/kg,RR 10-14 bpm,I:E 1:2) group,IPPV + positive end-expiratory pressure (PEEP) group and IPPV + continuous positive airway pressure (CPAP) group.Double-lumen tube was inserted.Conrrect positioning was verified by fiberoptic bronchoscopy.The patients were mechanically ventilated.PETCO2 was maintained at 35-45 mm Hg.In group IPPV + PEEP,OLV was performed for 30 min with PEEP of 5 cm H2O and then for another 30 min with PEEP of 10 cm H2O.In group IPPV + CPAP,OLV was performed with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side (for 1 h).MAP,HR,cardiac output (CO),cardiac index ( CI),stroke volume (SV),and stnoke volume index (SVI) were recorded before induction of anesthesia,at 10 min after intubation,at 30 min of two-lung ventilation,at 30 min and 1 h of OLV,and at the end of operation ( T1-6 ).Arterial blood samples were taken at T1,2,4-6 for blood gas analysis.The levels of blood glucose and lactate were measured.Oxygen delivery ( DO2 ) and DO2 index ( DO2I) were calculated.Results Compared with IPPV group,SV,SVI,CO,CI,DO2 and DO2I were significantly decreased at T4,5 ( P ＜ 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + PEEP,and no significant change was found in the parameters mentioned above in group IPPV + CPAP ( P ＞ 0.05).Compared with IPPV + PEEP group,SV,SVI,CO,CI,DO2 and DO2I were significantly increased at T4,5 ( P ＜ 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + CPAP ( P ＞ 0.05).Conclusion It exerts no influence on hemodynamics using OLV with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side,however,OLV with IPPV + PEEP can result in hemodynamic fluctuation,but the degree of fluctuation is lesser and DO2 can be maintained in the patients undergoing thoracic operation.

The morphology,cultural characteristics,physiological and biochemical properties of phosphate solubilizing strain BL-11 were studied.Combine with its 16S rDNA sequence analysis result strain BL-11 was identified as Brevibacillus laterosporus.Results showed that the phosphate solubilizing efficiency of strain BL-11was up to 10.91% when with Ca_ 3 (PO_ 3 )_ 2 as the sole phosphorus source in the medium,while the efficiency of that was up to 1.56% when with sands as the sole phosphorus source.An optimal media of BL-11 strain for solubilizing phosphate was obtained by orthogonal test.It was composed of sugar 20g/L,(NH_ 4 )_ 2 HCO_ 3 0.3g/L,MgSO_ 4 .7H_ 2 O 0.5g/L,NaCl 0.3{g/L},KCl 0.5g/L,FeSO_ 4 0.03g/L,MnSO_ 4 .H_ 2 O 0.03{g/L.}_ While other conditions were consisted of initial pH7.0-8.0,180 r/min,and 30℃.