Objective:To establish an HPLC method for the determination of related substances in enalapril maleate dispersible tablets.Methods:AKromasil100-SC8(250mm×4.6mm,5μm)columnwasused. ThemobilephaseAwas0.01mol·L-1potassi-um dihydrogen phosphate (adjusting pH to 2. 2 with phosphoric acid) and mobile phase B was acetonitrile with gradient elution,and the flow rate was 1. 0 ml· min-1 . The detection wavelength was 215nm,the column temperature was 40℃,and the injection volume was 20 μl. Results:The linear range of enalaprilat,enalapril maleate and eanlapril diketopiperazine was 14.64-43.92 μg·ml-1(r =0. 999 8), 9. 29-27. 87 μg·ml-1(r=0. 999 1)and 10. 80-32. 40 μg·ml-1(r=0. 999 5), respectively. Enalaprilat was not detec-ted out and enalapril diketone was 0. 2% using the method described in Chinese pharmacopeia, and the content of enalaprilat was 0. 1% and that of enalapril diketone was 0. 2% using the gradient elution. The related substances and enalapril maleate could be sepa-rated well. Conclusion:The method is sensitive, accurate and highly specific in the quality evaluation and control.

Objective:To explore the establishment and evaluation method of recurrent infection model of herpes simplex keratitis (HSK).Methods:To determine the optimal duration of the ultraviolet light irradiation, 12 healthy BALB/c mice were randomized into 3 minutes group, 2 minutes and 45 seconds group, and 2 minutes and 30 seconds group based on the duration of the ultraviolet light irradiation according to random number table method, with 4 mice in each group.Another 72 healthy BALB/c mice were randomized into blank control group, model group and recurrence group according to random number table method, with 24 mice in each group.The 72 mice were scratched a # symbol on their right corneas with scalpel.Then the eyes in the blank control group were treated with 5 μl of normal saline solution, while the eyes in the model group and recurrence group were treated with 5 μl of herpes simplex virus I (HSV-1) suspension.All the 72 mice were not intraperitoneally injected with HSV-1 xenogeneic serum antibody.Five weeks after the initial infection, the mice in the recurrence group were irradiated at ultraviolet B-302 nm for the optimal duration.The feasibility of establishing HSK recurrence model induced by this method was evaluated by the score of ocular surface symptoms, and the cytopathic effect (CPE) lesion analysis of Vero cells cultured in corneal wiping fluid from the mice in the model group and recurrence group.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO). The study protocol was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (No.PZSHUTCM190222039).Results:The optimal duration of the ultraviolet light irradiation was 2 minutes and 45 seconds.Within one week, HSK syndromes appeared in the mice injected with HSV-1 virus in the model group and recurrence group, and then gradually disappeared after one week.Before inducing recurrence, slit lamp examination was performed, and there was no spontaneous recurrence of HSK in the model group and recurrence group.The mice in the recurrence group relapsed within one week after ultraviolet light irradiation.The symptom scores of ocular surface lesions in the blank control group, model group and recurrence group were 0.333±0.471, 1.500±0.764 and 2.667±0.943 at one day after ultraviolet light irradiation, 0.000±0.000, 0.833±0.373 and 5.167±2.267 at three days after ultraviolet light irradiation, and 0.167±0.373, 1.000±0.577 and 3.000±1.155 at seven days after ultraviolet light irradiation, respectively.The symptom score of ocular surface lesions was higher in the recurrence group than the blank group and the model group at three days after ultraviolet light irradiation, and the differences were statistically significant (both at P<0.05). Morphological changes such as floating and gathering were found in the Vero cells after being cultured in corneal wiping fluid.The positive rate of CPE lesion was 71% (17/24) in the recurrence group, which was significantly higher than 8% (2/24) in the model group, and the difference was statistically significant ( P<0.01). Combining the two indicators, the success rate of recurrence in established models could reach 71%. Conclusions:Ultraviolet light irradiation can successfully induce the recurrence of viral keratitis in HSK mice without injection of neutralizing serum antibody.

OBJECTIVE:To study the protective effect of Compound pueraria tablets on the aging model rats. METHODS:The rats were divided into normal group,model control group,positive control group (vitamin E) and Compond pueraria tablets high-dose,medium-dose and low-dose (800,400,200 mg/kg) groups. The aging model rats were made by injection of D-galac-tose solution for 30 d in latter 5 groups,and they were given relevant medicine at same time every 7 days for consecutive 56 days since 31st day. SOD activity and MDA content in the serum and myocardium and LPF contents in the myocardium were determined after the last dosing,and apoptotic cells were counted and the pathology of cerebral tissues were observed. RESULTS:Compared with normal group,the activity of SOD in the serum and myocardium were decreased significantly in model control group,MDA and LPF contents in the myocardium and the number of apoptotic cells were increased significantly(P<0.01);significant myocar-dial cell injury was found. Compared with model control group,the activity of SOD in the serum and myocardium were increased significantly,while MDA and LPF content in the myocardium and the number of apoptotic cells in Compound pueraria tablets high-dose,medium-dose and low-dose groups were decreased significantly(P<0.05 or P<0.01);myocardial cell morphology was improved significantly. CONCLUSIONS:Compound pueraria tablets has obvious protective effect on myocardial cells in aging rats induced by D-galactose,the mechanism maybe related to the increase of SOD activity and the decrease of the content of MDA and LPF.

The blocking effect of selenium-enriched malt (Se-malt) on AFB1-in-duced UDS was reported. When the rats were fed with food containing lppm Se in form of Se-malt for two weeks, the UDS values of leukocytes treated with 6?10-7M AFB1 in vitro was decreased from 54.8 ?14.0 opm/106 WBC to 35.1?10.0 cpm/106 WBC (p

OBJECTIVE:To study the effects Weide'an tablet on gastric function and the expressions of epidermal growth fac-tor(EGF)and its receptor(EGFR)in gastric tissue of model rats with stress-induced gastric injury,and investigate its mechanism in the treatment of gastric ulcer. METHODS:Rats were randomly divided into normal group (normal saline),model group (nor-mal saline),positive group (ranitidine hydrochloride,0.015 mg/kg) and Weide'an tablet high-dose,medium-dose,low-dose groups (1.7,0.87,0.43 g/kg),10 in each group. Except for normal group,rats in other groups were given fasting swimming in cold water to induce gastric ulcer model. After modeling,all rats were intragastrically administrated once a day,for 2 weeks. The body mass of rats in 0,7,14 d of administration was recorded. After 12 h of last administration,gastric juice secretion,gastric juice pH,gastric ulcer area and gastric mucosal damage index of rats were detected,and the expressions of EGF and EGFR in gas-tric tissue were detected. RESULTS:Compared with normal group,body mass and gastric juice pH in 7,14 d in model group were declined;gastric juice secretion and gastric ulcer area were increased,gastric mucosal damage index was increased;and the expressions of EGF and EGFR in gastric tissue were enhanced,with statistical significances(P<0.05 or P<0.01). Compared with model group,except that the body mass,gastric juice secretion,gastric juice pH and gastric ulcer area in Weide'an medium-dose group in 7 d,and body mass,gastric juice secretion,gastric juice pH,gastric ulcer area and the expression of EGF in gastric tis-sue in Weide'an low-dose group in 7,14 d were not significantly improved,above indexes were significantly improved in other ad-ministration groups(P<0.05 or P<0.01);and the effect of Weide'an tablet showed a certain dose-effect relationship. CONCLU-SIONS:Weide'an tablet can significantly improve the gastric function of model rats with stress-induced gastric injury;and the mechanism may be related with enhancing the expressions of EGF and EGFR and promoting ulcer healing.

Objective To quantitatively assess the cardiopulmonary exercise function of spinal cord injury (SCI) patients and observe the effect of aerobic exercise on cardiopulmonary function, motor function and activities of daily living. Methods From December, 2014 to June, 2016, 34 incomplete SCI patients (ASIA C and D) and 23 healthy controls received cardiopulmonary exercise test (CPET). SCI pa-tients were randomly divided into conventional rehabilitation group (n=17) and aerobic exercise group (n=17). The aerobic exercise group received aerobic exercise for four weeks. They were assessed with CPET, motor and sensory function, walking index for spinal cord injury II (WISCI II) and spinal cord independence measure (SCIM) before and four weeks after training. Results Oxygen uptake (VO2)peak, anaerobic threshold (AT), metabolic equivalent of energy (METpeak), VO2/heart rate (HR)peak, respiratory exchange rate (RER)peak, minute ventilation (VE)peak, work rate (WR)peak and systolic blood pressure (SBP)peak were lower in the patients than in the controls (t>2.714, P<0.05). VO2peak、AT、METpeak、VO2/HRpeak、WRpeak increased in the aerobic exercise group after training (t>2.431, P<0.05). METpeak and WRpeak improved in the conventional rehabilitation group after training (t>3.282, P<0.01). The scores of motor in ASIA and SCIM improved in both groups after training (t>2.985, P<0.05). Conclusion The cardiopulmonary function decreased in incomplete SCI patients, which could be improved by moderate intensity aerobic exercise.

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.

AIM: G protein-coupled inwardly rectifier potassium (GIRK) channel are distributed widely in mammalian brain. In CNS, GIRK 1/2 seems to be the predominant heterotetramers which play a pivot role in the regulation of the excitability of neurons and may contribute to the resting potential by leading to a hyperpolarization of membrane potential and reduction of the action potential frequency. In the context, the Weaver mouse is the first neurological abnormality directly linked to a genetic point mutation in the GIRK2 protein which includes spontaneous seizure. GIRK2 knock out mice showed normal development but more susceptible than normal mice to seizure induced by GABA antagonist. Here, we report that the mRNA and protein expression of GIRK subunit 2 is altered in kainic acid(KA)-induced epileptic rat hippocampus. METHODS: Rats were injected with kainate 14 mg/kg intraperitoneally to establish an acute and chronic temporal lobe epilepsy model. At chronic spontaneous seizure stage, by using of in situ hybridization, immunocytochemistry and Western blotting, the GIRK 1,2 mRNA and protein were analyzed quantitatively in the dentate gyreus, CA1, CA3 regions of hippocampus. RESULTS: GIRK1,2 mRNA and proteins were expressed abundantly in all regions of hippocampus. KA induced seizures and caused a significant increase in GIRK2 mRNA abundance and immunoreacitivity; only GIRK1 mRNA was increased significantly, but no difference was found by Western blotting protocol. CONCLUSION: GIRK1,2 mRNA and protein expression in the hippocampus of epileptic rat brain is up-regulated, which may be an adaptive response to over-excitability of neuron networks and prevent the over-excitability spread in hippocampus (DG-CA3-CA1). [