Objective To investigate the expression and cellular localization of cyclin-dependent kinase 5 (Cdk5) in cerebral cortex after subarachnoid hemorrhage (SAH) in rats.Methods Fifty-two male Sprague Dawley (SD) rats were randomly divided into either a sham operation group (n =12) or a SAH group (n =40).The latter was randomly redivided into 6,12,24 h,and day 2 and 3 subgroups (n =8 in each group).A rat SAH model was induced by injecting fresh blood into the prechiasmatic cistern.Western blot and immunohistochemistry were used to detect the expression of Cdk5 in rat brain cortex.Double labeling immunofluorescence staining was used to detect the cellular localization of Cdk5 protein in cerebral cortex.Neuronal nuclear antigen labeled neurons,and glial fibrillary acidic protein labeled astrocytes.Results Western blot showed that the expression of Cdk5 protein was up-regulated at 12 hours after SAH (t =3.709,P =0.001),and it reached the peak on day 1 (t =3.475,P=0.002).Immunohistochemistry showed that the proportion of Cdk5 positive cell was also increased gradually after SAH,and the changes of time course were consistent with the results of Western blot,and it reached the peak on day 1 (t =4.320,P =0.000).Double labeling immunofluorescence showed that Cdk5 was mainly expressed in the neuronal cytoplasm in the sham operation group,and Cdk5 shifted to the neuronal nuclei in the SAH group.Cdk5 was mainly colocalized between astrocytes and neurons.Conclusions SAH up-regulates the expression of Cdk5 protein in cerebral cortex.Cdk5 may be involved in early brain injury after SAH.

Objective: To explore the risk factors in patients with chronic heart failure (CHF) combining atrial ifbrillation (AF) and to identify the predictive factors for AF recurrence after cardio version. Methods: A total of 246 CHF patients treated in our hospital were divided into 2 groups: CHF+AF (AF) group,n=71 and CHF without AF (Non-AF) group,n=175. The baseline information was recorded. The risk factors for AF occurrence were screened by Logistic regression analysis. There were 66 AF patients with successful cardio verse and they were followed-up for 18 months, the predictors for AF recurrence were studied by Cox regression analysis. Results: The baseline information showed that age, CHF duration, left atria diameter (LAD), the levels of BNP, uric acid and serum creatinine in AF group were all higher than those in Non-AF group, allP<0.05. Logistic regression analysis presented that age (RR=1.89, 95% CI 1.08-3.31,P<0.05), CHF duration (RR=1.35, 95% CI 1.05-1.74,P<0.05), BNP level (RR=1.82, 95% CI 1.30-2.53,P<0.01), LAD (RR=1.51, 95% CI 1.09-2.08,P<0.05) were the risk factors for AF occurrence. Cox regression analysis indicated that BNP level (RR=1.30, 95% CI 1.09-1.54,P<0.01) and LAD (RR=1.74, 95% CI 1.30-2.34, P<0.01) were the predictors for AF recurrence. Conclusion: The elder age, longer CHF duration, increased BNP level and LAD were the risk factors for AF occurrence in CHF patients. The increased BNP level and LAD were the objective predictors for AF recurrence after cardio version.

Objective To study the anatomical basis and clinical causes of splenic infarction with an attempt to improve on the diagnosis and treatment of splenic infarction.Methods This study was conducted on 11 patients with splenic infarction seen in our hospital from December 2003 to September 2012,131 patients with a clinical diagnosis and treatment reported in the literature since 1999,and 25 adult cadavers showing the anatomy of the splenic arteries with an aim to find out the causes of splenic infarction.Results The clinical data showed that splenic infarction occurred more commonly in patients 60 years of age or older (73％ vs 27％,P＜0.05),and in males more than in females (62％ vs 38％,P＜0.05).The diagnosis was first made significantly more often by physicians than by surgeons 88/32 (79％ vs 21％,P＜0.05).The anatomical data showed that the majority of the splenic arteries in the 25 adult cadavers was curved.Conclusions The clinical manifestations of splenic infarction easily led to a misdiagnosis.Improvement in the diagnosis and treatment of splenic infarction would depend on the clinical awareness of this condition,the prothrombotic state detection and the implementation of timely and standardized treatment.

BACKGROUND:Pueraria is a member of isoflavone characterizing by decreasing the toxic effects of alcohol,relieving the effect of alcohol withdrawal,anti-oxidative effect,removing oxygen free radicals,and preventing cell injury induced by lipid peroxidation.OBJECTIVE:To investigate the inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells (hBMSCs).DESIGN,TIME AND SETTING:An in vitro observation based on cytology was performed at Laboratory of Biochemistry and Molecular Biology,Basic Medical College of Zhengzhou University in October 2008.MATERIALS:The bone marrow was taken from the adult healthy volunteers from the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University.Puerarin standard was provided by the National Institute for the Control of Pharmaceutical and Biological Products.METHODS:The primary hBMSCs were purified from the adult marrow by gradient centrifugation.The second passaged cells were divided into 3 groups.Blank control group:Cells were cultured with DMEM culture media containing 10% fetal bovine serum;Model group:Cells were cultured with 0.09 mol/L alcohol;Experimental group:Cells were cultured with 0.09 mol/L alcohol and 10-6 mol/L puerarin;while,the culture liquid was changed every two or three days.Cells were cultured for 7 days in each group.MAIN OUTCOME MEASURES:PPARγ 2 gene expression with RT-PCR;amount of adipocyte with oil red "O" staining;alkaline phosphatase activity.RESULTS:PPAR-γ2 mRNA expression in the model group was significantly greater than experimental group and blank control group (P ＜0.05),while the expression in the experimental group was slightly greater than blank control group,but there was no significant difference (P ＞0.05).The number of adipocytes was the most in the model group,and the lipid droplet was large and abundant.The number of adipocytes in the experimental group was significantly less than model group (P ＜ 0.05),and there were few adipocytes in the blank control group.Compared with blank control group,alkaline phosphatase activity was decreased in the model group (P＜0.05);but the activity in the experimental group was increased compared to model group (P ＜ 0.05).CONCLUSION:Puerarin inhibits differentiation of hBMSCs into adipocytes induced by alcohol.

Objective To explore the common pathogen of infantile convulsions associated with mild gastroenteritis, and to study the differences between the seizures caused by the two kinds of virus.Methods RT-PCR was used to detect Rotavirus (RV) and Norwalkvirus (NoV) in stool and cerebrospinal fluid of 30 cases with infantile convulsions associated with mild gastroenteritis. The differences between the frequency of seizures caused by two kinds of virus were analyzed by statistical methods (two-sample t-test).Results 17/30 (56. 7%) were RV-positive in stool and 3/17 (17. 7%) in cerebrospinal fluid; 6/30 (25.0%) were NoV-positive in stool and 1/6 (16. 7%) in cerebrospinal fluid. The seizure frequency with NoV infection was (4. 33 ± 1.75) times, and RV infection patients was (2. 53 ± 1.12) times (P ＜ 0. 01).The seizure frequency of CSF virus-positive children was (4. 75 ± 1.71) times compared to (2. 63 ± 1.21)times in virus-negative children (P ＜ 0.01). Conclusion The common pathogens causing infantile convulsions associated with mild gastroenteritis were RV and NoV. The degree of NoV infection affecting the central nervous system may be greater than RV. The presence of the virus in cerebrospinal fluid may lead to higher incidence of seizures,but their exact roles related to the occurrence of seizures remain to be further studied.

Objective The present study was designed to investigate changes in serum or plasma concentrations of nitric oxide and its derivatives in diabetic patients.Methods Serum nitrate concentration of 84 diabetic patients was measured by using an enzyme kinetic method,and the plasma S-nitrosothiols concentration of 10 cases was measured by using HPLC technique.Results Serum nitrate concentration and plasma S-nitrosothiols concentration in the diabetics were significantly higher than in control group (P＜0.01 andP＜0.05,respectively).The serum nitrate concentration in diabetics also had a significant positive correlation with the serum glucose concentration (R=0.7256,P＜0.05),but this correlation was not found in control group.Conclusion These data showed that NO and its derivatives are overproduced in the diabetic patients.(J Geriatr Cardiol 2008;5:25-27)

Objective To observe the effect of sterimar physiological sea water spray nasal spray on children with mild intermittent allergic rhinitis (AR).Methods 81 cases with mild intermittent AR from March 2014 to March 2015.All the cases were divided into observation group (n=40) and control group (n=41) according to the double blind and random number table method.The control group were given conventional loratadine treatment, observation group on the basis of the control group given sterimar physiological sea water spray nasal spray treatment.The symptoms of allergic rhinitis, signs score, mucociliary transport rate and clinical curative effect between two groups pre-and post-treatment were analysed and compared.Results After treatment, the symptoms of allergic rhinitis, signs score in observation group were significantly lower than those in control group (t=12.908,t=11.340;P<0.05).The mucociliary transport rate in observation group was significantly higher than that in control group (t =4.119;P<0.05).The total effective rate of observation group was significantly higher than that of control group (97.50%vs.80.49%;χ2 =5.933,P<0.05).Conclusion The clinical treatment effect of sterimar physiological sea water spray nasal spray could effectively alleviate the symptoms in the treatment of children with mild intermittent AR, which has an accurate efficacy.

Objective To investigate the feasibility of gene modification of peripheral blood derived endothelial progenitor cells(EPCs).Methods The rabbit mononuclear cells were collected from rabbit peripheral blood and cultured.The uptaking test of DiI-Ac-LDL was directly visualized with fluorescent microscopy,the immunofluorescence analysis was performed to detect the expression of surface marker.And the human tissue kallikrein(HTK)gene was amplified with PCR,and inserted into pEGFP-N3 vector.EPCs were transfected with the constructed plasmid by means of lipidosome,and the HTK-EGFP fused protein was visualized directly with fluorescent microscopy,and the expression of HTK was detected by RT-PCR and ELISA.Results After 5～6 days of culture,spindle-shaped attached cells clustered together,and cobblestone-like cell layer appeared after 15 days.Cells were positive of uptaking DiI-Ac-LDL and expressing vWF and CD133 related antigen.Plasmids were correctly formed and expressed in the rabbit EPCs.The fused protein had activity of both HTK and EGFP.Conclusion The HTK gene modified EPCs is potential in the treatment of artery injury and endothelial dysfunction.

ObjectiveThis clinical study is aimed to investigate whether levetiracetam (LEV) can improve electrocorticogram (EEG) in epileptic children epilepsy patients with better clinical manifestation but abnormal EEG findings.MethodsTotally 39 children from our neurological clinic with partial or complex partial epilepsy seizure were included in present study and assigned equally into three groups receiving different treatment:control group,sodium valproate (VPA) group,and LEV group.Their clinical symptoms had been controlled for over one year by carbamazepine ( CBZ),but EEG results showed clearly abnormal.Epileptiform discharges were observed in routine EEG exams half a month before recruiting.After recruiting,they continued to receive CBZ alone (control group) or co-treated with VPA ( VPA group) or LEV (LEVgroup),respectively.Six months later,EEG was taken again and results were analyzed.ResultsImprovement rate were 9.1％ ( control group),23.1％ ( VPA group),and 66.7％ (LEV group),respectively;Overall statistical difference was reached among three groups ( P＜0.01 ) and between control group and LEV group( P＜0.012 5 ),but no statistical difference between control group and VPA group was reached ( P＞0.0125).ConclusionCo-treatment of LEV in child epilepsy patients receiving CBZ can significantly decrease abnormal EEG discharge frequency during interictal period.

BACKGROUND:Several reports have demonstrated that metabolic disorders and physiopathologic changes accompany with urinary diversion.But these metabolic disorders caused by bladder reconstruction using intestinal tract are related to type and length of intestinal canal.OBJECTIVE:To investigate the histological change of reservoir mucosa and to assess effects on metabolic state in patients with a Roux-y sigmoid neobladder.DESIGN,TIME AND SETTING:A retrospective case analysis was performed at the Department of Urinary Surgery,the 184 Hospital of Chinese PLA between June 2000 and November 2008.PARTICIPANTS:The experimental group comprised 33 bladder carcinoma patients,21 males and 12 females,averaging 64 years of age.The control group consisted of 25 subjects who had no sigmoid colon diseases confirmed by gastroenterological endoscopy.METHODS:Patients with bladder carcinoma received radical cystectomy and bladder reconstruction using Roux-y sigmoid neobladder which controlled urination with anal sphincter.Prior to and after neobladder drainage tube removal,serum levels of electrolyte,creatinine,and urea nitrogen were detected.Before and 36 months after surgery,reservoir mucosa from 13 patients with bladder carcinoma was pathologically examined.For the control group,the thickness of sigmoid colon mucosa and the numbers of intestinal glands were determined.MAIN OUTCOME MEASURES:Electrolyte,renal function,acid-base balance,mucosal layer thickness,numbers of intestinal glands prior to and after surgery,as well as prior to and after drainage tube extraction.RESULTS:After surgery,electrolyte,creatinine,and urea nitrogen were all normal in 30 patients.There was no significant difference in serum electrolyte,creatinine,and urea nitrogen between prior to and after surgery.Mild acid poisoning was found in 3 patients.Microscopic observation results revealed that sigmoid colon mucosa in the control group did not change significantly after surgery,and it basically kept the normal tissue structure;in the experimental group,sigmoid colon mucosa that was(577.6±169.4)μm prior to surgery was thinned(412.5±114.7)μm(P＜0.05),intestinal glands were loosely arranged,interstitial substance became less,and the number of intestinal glands per high-fold visual field that was(26.4±3.5)/high-fold visual field prior to surgery was decreased(15.2±2.7)/high-fold visual field(P＜0.05),after surgery.In addition,intestinal villus in the neobladder was gradually atrophied,and no enterocyte proliferation and malignant changes were found after surgery.CONCLUSION:After Roux-y sigmoid neobladder application,colon mucosa was gradually thinned,intestinal glands were loosely arranged,interstitial substance became less,the number of glands per high-fold visual field was decreased,and body metabolism produced no changes.