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Objective: To investigate the mechanisms responsible for the neuroprotection by cholecystokinin octapeptide against glutamate-induced apoptosis in vitro cultured cortical neurons. Methods: Primary cultured corticaI neurons from SD rats of 0～24 hold were incubated for 8 days. The cultured cells were divided randomly into three groups: control group,glutamate group and CCK group. In the controI group,cells were not treated with glutamate orCCK;Neurons in glutamate group were incubated with 50?mol/Lglutamate for 30 min;In CCK group,CCK-8 was added to the Neurons 24 h prior to incubation with glutamate. After injuried by glutamate,cells in all the groups were incubated with normal medium for 0,6,12,24 h and 48 h. At the five time points,cells were fixed respectively for experiment. Cell viability were determined by the colorimetric MTT assay;The protein expression of Bcl-2,Bax and Caspase-3 were determined by immunocytochemistry techniques. Results:Pretreatment with CCK for 24 h significantly improved glutamate-induced suppression of cell viability. Pretreatment with CCK also completely reversed the suppression of Bcl-2 expression,and significantly inhibited Bax overexpression and Caspase-3 activition induced by glutamate. Conclusion:Theneuroprotective mechanisms of CCK against glutamate-induced apoptosis in cultured cortical neurons may be associated with up-regulation of Bcl-2/Bax ratio and down-regulation of Caspase-3.

Objective To explore renal clear cell carcinoma ( CCRCC) expression and significance of CD146 mR-NA in tumor tissue of patients with. Methods ELISA method for quantitative detection was used for CD146 protein and real-time PCR technique for the detection of CCRCC in 102 ( CCRCC) expression in tumor tissue of patients with CD146 mRNA, and 51 cases of renal patients with non tumor tissues as control. Results Found metastasis in patients with CCRCC, the CD146 protein concentration was statistically significant compared with the control group (F=52.1, P<0.01),the average expression of CD146 mRNA value (0.043 8±0.002 4) was significantly high-er than that of in situ CCRCC patients (0.038 2±0.001 1, P= 0.018) and control group (0.034 4±0.001 0, P=0.001 ) . Conclusion Pathological grading and lymph node up regulation of CD146 mRNA expression in renal cell carcinoma and metastasis, is expected to become a new index, evaluation of malignant degree of CCRCC me-tastasis and prognosis, and provide a reliable basis for the intervention of clinical treatment.

Objective To investigate the clinical significance of the joint detection of platelet parameters and coagulation function markers in hepatitis patients.Methods 310 patients with viral hepatitis were divided into 4 groups:acute hepatitis group,chronic hepatitis group,serious hepatitis group and hepatocirrhosis group.The control group was the 80 healthy examination eligible people.Each group were detected platelet parameters ( PLT), mean platelet volume( MPV),.platelet distribution width( PDW),platelet large cell ratio( P-LCR), prothrombin time( PT) ,activated partial thromboplasin time (APTT ) , thrombin time ( TT) and factorl ( Fbg).Results The levels of PLT,MPV and Fbg were obviously reduced in chronic hepatitis,serious condition chronic hepatitis patients and hepatocirrhosis had significant difference compared with those in the control group (all P < 0.01).Moreover, the levels of PDW,P-LCR,PT,APTT and TT were obviously increased in the four groups compared with the control group(all P <0.05 ).Conclusion The joint detection of platelet parameters and coagulation function markers could help to observe the degree of progression of virus hepatitis,and had important clinical significance for the treatment and prognosis.

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Objective To study the pathological spectrum of neonatal sepsis and antibiotic resistance of bacteria and fungi isolated from neonatal blood samples. Methods Clinical information of neonates with sepsis admitted to neonatal ward of our Hospital were retrospectively collected from January 2007 to December 2014 and analyzed. Results During the study period, a total of 23 224 infants were admitted to neonatal ward. Among them, 608 were diagnosed with sepsis (2. 62% , 608 / 23 224). Blood culture samples were positive in 412 cases, the rest (196 cases) were diagnosed according to clinical manifestations and laboratory tests. The composition ratio of sepsis during the first 4 years was significantly lower than that of the subsequent 4 years (2. 58% , 225 / 9 805 vs. 2. 85% , 383 / 13 419, P < 0. 05). Of 412 bacterial strains isolated, gram-positive bacteria, gram-negative bacteria and fungi accounted for 62. 38% , 30. 34% and 7. 28% , respectively. For the 155 strains isolated during first 4 years, the number of G + bacteria, G - bacteria and fungi were 106 strains (68. 39% ), 40 strains (25. 81% ), 9 strains ( 5. 81% ), respectively. The coagulase-negative staphylococcus ( CONS ) accounted for 90. 57% of G + strains. Klebsiella pneumoniae and Escherichia coli accounted for 50. 00%and 15. 00% of G - strains respectively. 257 strains were isolated during the subsequent 4 years, of which G + bacteria, G - bacteria and fungi accounted for 151 strains (58. 75% ), 85 strains (33. 07% ), 21 strains (8. 17% ), respectively. CONS accounted for 74. 83% of G + strains, Klebsiella pneumoniae and Escherichia coli accounted for 30. 58% and 35. 29% of G - strains. The composition ratio of CONS and Escherichia coli were significantly different between first and subsequent 4 years (P < 0. 05). G +bacteria were resistant to penicillin, oxacillin and macrolide, sensitive to piperacillin / tazobactam and quinolone. We did not identify any G + strain resistant to vancomycin. In general, G - bacteria were resistant to ampicillin and cephalosporins, sensitive to the beta lactamase inhibitor compound preparation and quinolones, and highly sensitive to carbapenems. Fungi generally were sensitive to conazoles. Conclusions Neonatal sepsis are mainly caused by CONS, Klebsiella pneumoniae and Escherichia coli also play important roles. G + bacteria and G - bacteria in general are resistant to penicillin and cephalosporins. All G + bacterial strains isolated from our cohort are sensitive to vancomycin. G - bacteria are generally sensitive to carbapenems. Fungi generally are sensitive to conazoles.

Objective To investigate the correlation between hepatitis B virus (HBV)drug -resistance gene mutations and hepatocellular carcinoma (HCC).Methods The clinical data of treatment -experienced patients,who underwent examination for HBV drug -resistance gene mutations in Beijing Ditan Hospital from January 1 to December 31,2013,and still had detectable HBV DNA after being treated with nucleos(t)ide analogues,were collected.All the patients were followed up,and the development of HCC was considered as the clinical out-come.The correlation between drug -resistance gene mutations and the development of HCC in patients with HBV infection was analyzed. The chi -square test was used for comparison of categorical between groups,the t -test was used for comparison of continuous data between two groups,and the log -rank test was used for comparison of the incidence of HCC between two groups.Results A total of 227 patients were enrolled in this study.According to the results of the detection of HBV drug -resistance gene mutations,103 patients (103 /227, 45.37%)had no drug -resistance gene mutations and 124 (124 /227,54.63%)had drug -resistance gene mutations.There were no sig-nificant differences between the mutation group and the non -mutation group in HBV DNA load (5.19 ±1.60 log10 IU /ml vs 5.44 ±1.75 log10 IU /ml,t =-1.134,P =0.258)and the percentage of patients with liver cirrhosis (24.19% (30 /124)vs 16.50% (17 /103),χ2 =2.026,P =0.155).The median follow -up was 28 months (range 4 -58 months),and the incidence of HCC was 7.49% (17 /227).A-mong the patients with HBV drug -resistance gene mutations,12 (12 /124,9.68%)developed HCC,and among those without HBV drug-resistance gene mutations,5 (5 /103,4.85%)developed HCC.Among the patients who developed HCC,70.59% (12 /17)had HBV drug -resistance gene mutations at baseline;among the patients who did not develop HCC,53.33% (112 /210)had HBV drug -resistance gene mutations at baseline.Conclusion The patients with poor control of HBV DNA during antiviral therapy have a comparable incidence of HCC to those not treated with antiviral therapy,with a relatively high risk of developing HCC;the treated patients with HBV drug -resist-ance gene mutations may have a higher risk of HCC than those without such mutations,which needs to be confirmed by the studies with a longer follow -up period and a larger sample size.

OBJECTIVE ToevaluatetheeffectandmechanismofPaecilomyceshepialimycelium
(PHM)onexhaustedfatigueinmice.METHODS Micewereforcedtorun30minadayforconsecutive 18 d using the wheel running fatigue apparatus, and the speed was gradually increased from 10.2 m·min -1 to 12.6 m·min -1 .On d19,these animals were forced to run 60 min to establish an exhausted fatigue model.Every day,40 min before the run training,the animals were ig administered with PHM (140,280,560 mg·kg -1 )and modafinil (13 mg·kg -1 ).The number of electric shocks was recorded on d5,d9,d1 3,d1 7 and d1 9,respectively,and the body mass of each mouse was recorded daily.The levels of muscle glycogen (MG),liver glycogen (LG),serum urea nitrogen (SUN),serum lactic acid (SLA)and serum creatine kinase (CK)were detected by commercially available kits. RESULTS Duringthelastexhaustedrunningond19,thenumberofelectricshocksofthemodelgroup was markedly larger than that of the drug administration groups.When compared to the normal control group,the model group showed a significant decrease in the level of either MG or LG,but a significant increase in the level of either SUN or CK.The nu mber of electric shocks of the PHM in the moderate and high dose groups was 133 ±55 and 1 15 ±41 times,respectively,which was significantly smaller than that of the model group 240 ±89 (P<0.01 ).The levels of MG and LG of the PHM560 mg·kg -1 group were (1 .28 ±0.24)mg·g -1 and (40.9 ±1 0.2)mg·g -1 ,respectively,which were significantly higher than those of the model group〔MG:(0.91 ±0.26)mg·g -1 ,LG:(22.2 ±2.9)mg·g -1;P<0.01〕,and the levels of SUN,SLA and CK of this PHM group were (5.8 ±1 .3)mmol·L-1 ,(5.7 ±1 .7)mmol·L-1 and (0.6 ±0.2)kU·L-1 ,respectively,which were significantly lower than those of the model group 〔SUN:(7.5 ±2.1 )mmol·L-1 ,SLA:(7.1 ±1 .8)mmol·L-1 ,CK:(0.8 ±0.3)kU·L-1;P<0.05〕.The number of electric shocks of the modafinil group was also significantly smaller than that of the model group (P<0.01 ),but there were not any significant differences between the modafinil group and the model groupinthelevelsofMG,LG,SUN,SLAorCK.CONCLUSION ThisstudyshowsthatPHMcanhelp relieve fatigue,which might be associated with its abilities to increase the storeage of energy sources,to enhance the aerobic metabolis m,and to decrease the accu mulation of metabolic products.

BACKGROUND: Hepatocyte transplantation can improve biochemical parameters and survival of animals with acute liver failure. However, cell sources, immunological rejection, distribution and histomorphological alternation of transplanted hepatocytes are important issues for its wide application in clinic. OBJECTIVE: To observe early-stage histomorphological and ultrastructural changes of SV40LT antigen gene transfected hepatocytes after transplanted into rat spleen. DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at Laboratory of National Hepatobiliary and Enteric Surgery Research Center, Ministry of Health from March to December 2001. MATERIALS: Sixty male Wistar rats were selected for hepatocyte transplantation. METHODS: Sixty rats were randomly divided into 4 groups (n=15). Primary cell group and primary cell plus cyclosporine A group were intrasplenically transplanted with primarily cultured hepatocytes; SV40LT antigen gene group and SV40LT antigen gene plus cyclosporine A group were intrasplenic transplanted with SV40LT antigen gene transfected hepatocytes. Twenty-four hours before and 14 days after transplantation, the rats in primary cell group and SV40LTor antigen gene group were injected with 0.5 mL normal saline through tail vein daily, while the other groups were injected with cyclosporine A (10 mg/kg per day). MAIN OUTCOME MEASURES: The spleen of one rat was harvested every day postoperatively for light microscopic and electron microscopic examinations to observe survival rate, histomorphological and ultrastructural features of transplanted hepatocytes for 14 days. RESULTS: Compared with primary cell group and SV40LT antigen gene group, the survival rate of transplanted hepatocytes in the other groups was significantly increased (P 0.05), while the survival rate in SV40LT antigen gene transfected hepatocytes was significantly higher than primarily cultured hepatocytes 8-14 days postoperatively (P

Objective To construct the eukaryotic expression vector of HBEBP2 gene,and screen the exonic genes of proteins in hepatocytes interacting with HBEBP2.Methods The DNA fragment of HBEBP2 was amplified by PCR.The eukaryotic expression vector pGBKT7-HBEBP2 was constructed successfully by yeast-two hybrid system 3 and then transformed into yeast cells AH109.The transformed AH109 were then mated with yeast cells Y187 containing hepatic cDNA library plasmid.The diploid yeast cells were plated on synthetic dropout nutrient medium(SD/-Trp/-Leu/-His/-Ade) containing X-?-gal for the first selection.Library plasmids pACT2-DNA were extracted and co-transformed into yeast cell AH109 together with pGBKT7-HBEBP2.Then the yeast cells were plated on synthetic dropout nutrient medium(SD/-Trp/-Leu /-His/-Ade) containing X-?-gal for the second screening to eliminate false positive clones.The real positive clones were sequenced and analyzed by bioinformatics.Results Six proteins binding to HBEBP2 were screened,including human sapiens lactate dehydrogenase D,human sapiens mitochondrion,human sapiens mannose,human sapiens aldehyde oxidase,human sapiens serpin peptidase inhibitor and other two unknown proteins.Conclusions A novel class of proteins in hepatocytes interacting with HBEBP2 has been obtained.It is presumed that HBEBP2 protein is correlated with glycosylation,lipid metabolism and cell proliferation,etc.