In order to identify whether functional L-type calcium channels are expressed in neural stem cells(NSCs) from rat embryonic hippocampus, and whether L-type calcium channels participate in the modulation of proliferation and differentiation of NSCs, the rat embryonic hippocampal tissue was dispersed into a single cell suspension, and the dissociated cells were cultured in serum-free DMEM/F12 medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), N-2 and B27 supplement. Immunofluorescent labeling showed an expression of nestin-positive cells. Following five-days culture in differentiation medium, neuron-like and astrocyte-like cells were observed, which expressed ?-tubulin Ⅲ(Tuj1) and Glial fibrillary acidic protein (GFAP), respectively. Western blot analysis showed an expression of Cav1.2?1C subunits in NSCs, but no Cav1.3?1D subunits. Moreover, L-type calcium channel currents were recorded in those cells by using whole-cell patch clamp techniques. It was found that activation of L-type calcium channels promotes proliferation and differentiation to neuronal type of NSCs. The results indicated that rat embryonic hippocampal NSCs express functional L-type calcium channels, L-type calcium channels modulate proliferation and differentiation of NSCs.

To propose the hypothesis of the requirement for optimum ratio of various active cons tituents of Chinese materia medica with desired effects, which suggests that both single drug and compound drug prescriptions have their own target effects corresponding to their main indications and their active constituents must be in optimum ratio This hypothesis was initially tested in an ischemic animal model The neuroprotection of Yushen Oral Liquor (YSOL, an oral liquor made of Chinese materia medica) on neuronal injury in CA1 region of rat hippocampus following transient forebrain ischemia was used as an index to evaluate the drug's effect By combination of HPLC and pharmacological method, the relationship of HPLC and pharmacodynamical results of each sample made of YSOL was studied to observe the chromatogram peaks representing the drug's neuro protective effects There are three peaks named as R1, R2, and R3 for YSOL, which closely correlate to the drug's neuronal protective effects and the relative weight but not the absolute quantity of those peaks plays a critical role in producing neuronal protective effects of YSOL on ischemia induced neuronal damage The results provide experimental data supporting our proposed hypothesis

The modulating action and mechanism of endogenous nitric oxide (NO) on the delayed rectifier potassium currents in cultured hippocampal neurons were examined using whole-cell patch clamp techniques. L-arginine (L-Arg, 2 mmol/L), a substrate of NO synthases, significantly suppressed the delayed rectifier K+ currents in hippocampal neurons, while its isomer D-arginine (D-Arg, 2 mmol/L) exerted no effect. Moreover, pretreatment with NO synthase inhibitor L-NAME (0.5 mmol/L) completely blocked the suppressing effect by L-Arg, indicating that L-Arg exerted its modulation by producing NO but not by itself. No effect was found on the L-Arg-induced inhibition by 10 min pretreatment of 10 |?mol/L ODQ (a specific inhibitor of guanylate cyclase). In contrast, thiol-alkylating agent N-ethylmaleimide (1 mmol/L) completely precluded L-Arg-induced inhibition on the whole K+ currents. The results indicate that endogenous NO modulates the delayed rectifier K+ currents in cultured hippocampal neurons mostly through S-nitrosylation.

OBJECTIVE To explore the clinical characteristics of nosocomial infection of lower respiratory tract by coagulase negative staphtylococci(CNS) and their drug resistance to common antibiotics.METHODS The clinical data and drug resistance of CNS of 37 nosocomial infection cases and 68 cases with colonization of lower respiratory tract by CNS were analyzed retrospectively,and the study mainly was focused on the risk factors and basic clinical features of nosocomial infection of lower respiratory tract by CNS.The resistance to common antibiotics of CNS was also summarized.RESULTS The risk factors were determined,which included long duration,age more than 65 years old,chronic obstructive pulmonary disease,the use of antibiotic suchs as carbapenems,the third generation cephalosporin,fluoroquinolones and concurrent infection of fungus.The mortality rate of infection was higher than after colonization.The clinical characteristics of nosocomial infection of lower respiratory tract by CNS included that the temperature of patients was focus on 37.5-38.5 ℃,peripheral blood routine of patients appearsed normal mostly and the lung imaging displayed the bilateral exudation changing near 50% the total.The 105 isolates of CNS were resistant to multi-antibiotics,the drug resistance rate to rifampicin and sodium fusidate was relatively low and resistant strains to vancomycin and teicoplanin were not detected.CONCLUSIONS The toxemia symptoms of nosocomial infection of lower respiratory tract by CNS are mild and the clinical manifestation is atypical.It is more likely to catch nosocomial infection of lower respiratory tract by CNS for the patients who are with low immunity and applied with broad spectrum antibiotic for long-term.

Objective: To investigate the changes of gallbadder motive function in diabetics. Method: The size of gnllbadder,recovering rate,echo and half-empty time after fat food were compared and statistically analyzed between 22 healthy individuals and 20 diabetics with ultrssonograph. Results:The gallbadder recovering rate was similar between the healthy individuals and diabetics (P>0.05), but the rate of cholecystitis (30%) and choletethiasis (35%) of diabetics were higher than those in healthy individuals, the gallbadder half-empty time after fat food was obviously slower in diabetics (46.1+12.9 min)than those healthy individuals (27+4.4 mmin). Conclusions: The diabetics had obvious gallbadder motive funciion obstruction and higher rates of chronic cholecystitis and choletethiasis with ultrasonograph observations.

Objective To investigate the relationship between nuclear factor(NF)-κB activity and lactacystin induced prostate cancer cell apoptosis.Methods Two prostate cancer cell were divided into two groups:blank control group treated with culture solution,lactacystin group treated with different concentration of lactacystin(0.5,1.0,2.0,4.0 μ mol/L),the action time were 8,16 and 24 hours.The cell survival rate was measured by MTT assay.NF-κB DNA binding activity was measured by enzyme-linked immunosorbent assay,the expression of NF-κB P65 nuclear protein was detected by Western blot assay,and caspase-3 activity was analyzed by enzyme analysis assay.Results On basal condition,the NF-κ B DNA binding activity was much higher in DU145 cell than that in LNCaP cell(t=4.728,P=0.001).Compared with blank control group,different concentration of lactacystin groups'NF-κ B DNA binding activity in both the LNCaP and DU145 cell were reduced.The expression of NF-κB p65 nuclear protein decreased along with raising of lactacystin concentration in LNCaP cell,but it did not change in DU145 cell.On basal condition,caspase-3activity in DU145 cell was higher than that in LNCaP cell(t=4.519,P=0.001).After lactacystin acting of 24 hours,caspase-3 activity increased along with raising of lactacystin concentration in both the LNCaP and DU145 cell(2.0 μmol/L lactacystin group compared with 1.0 μmol/L lactacystin group,DU145 cell P=0.000,LNCaP cell P=0.000).Conclusions Lactacystin has different killing effects on prostate cancer cell.The mechanism may be related to inducing the apoptosis by down-regulation of NF-κB activity.There may be additional cell survival/death pathway in androgen-independent prostate cancer cell.

The computer-based case simulation is a method of medical exam which can test students to solve the clinical ability,in particular the clinical policy-making thought and it is one of important methods of solving patient problem ability.This article is to introduce concept,the development as well as scoring method.It also expounds the superiority of the computer-based case simulation and shortcoming of the scoring method to provide the warrenty for the application in the nation-wide range in future.

Objective To use ELISA detecting self‐developed hepatitis B five internal quality control products .Methods Hepa‐titis B five positive sera by the detection of ELISA were diluted through optimal ratio ,and were homemade indoor quality control materials .Results Self‐control materials and commodities were simultaneously detected by ELISA ,and the test results were com‐pared ,the two were no significant difference(P>0 .05);Self‐control materials continuously detected by ELISA ,its batch variation were less than 15% ,and stability was in line with the requirements .Conclusion Self‐developed hepatitis B five indoor quality con‐trol materials are made simply ,have good stability ,are satisfied control effect ,and have promotional value .

Objective To investigate the resistance-mechanism of the carbapenems-resistant Klebsiella pneumoniae isolated from clinical.Methods The clinical isolates of carbapenems-resistant Klebsiella pneumoniae from top three comprehensive hospitals of Nanjing area were examined by 40 beta-lactamase,porin-coding genes and linkage of KPC-ISKpn6 using PCR method,the PCR positive results were picked out for sequencing and sequencing BLAST search for comparison analysis.Results Twenty-four strains of carbapenems-resistant Klebsiella pneumoniae were detected,the positive rate of A beta-lactamase TEM-1 and SHV was 100％ (24/24),KPC-2 and LAP-2 was 95.8％ (23/24),45.8％ (11/24) respectively,and C beta-lactamase DHA was 4.2％ (1/24).Meanwhile,the positive detection rates of KPC-ISKpn6 linkage was 95.8％ (23/24),and the mutation rate of porin-coding genes ompK35 and ompK36 were up to 95.8％ (23/24) and 100％ (24/24).Conclusion High incidence of beta-lactamase TEM-1,SHV,KPC-2 and LAP-2 was found in the group of Klebsiella pneumoniae isolates,and carbapenems-resistant of which was primarily due to the high carrying rate of KPC-2 and the high mutation rate of porin-coding genes ompK35 and ompK36.The Insertion sequence ISKpn6 may be involved in the KPC-2 gene mediated-expression.

Objective To estimate the safety and feasibility of fast track surgery (FTS) in operations for gastric cancer.Methods The Cochrane library,PubMed,EMBASE,Chinese biomedical literature database,China journal full text database,Chinese scientific journals full text database were searched with the key words of gastric cancer,gastric carcinoma, FTS,fast track surgery, enhanced recovery after surgery,ERAS, 胃癌,and 快速康复外科.The search was finished in April,2011.Randomized controlled trials of FTS for patients with gastric cancer were included.Two reviewers evaluated the quality of the included studies and extracted the data independently.Meta analysis was performed by RevMan 5.1.2 software.Results Fourteen randomized controlled trials involving 1270 patients were included.The results of meta analysis showed that FTS could significantly shorten the first exhaust time (MD =-6.15,95 ％ confidence interval:-8.22- -4.07,P＜ 0.05) and hospital stay ( MD =- 2.68,95％ confidence interval:- 3.26- - 2.10,P ＜ 0.05 ),and reduced the hospital fees ( MD =- 0.39,95 ％ confidence interval:- 0.55- - 0.22,P ＜ 0.05 ).FTS could lighten the postoperative stress and relatively enhance the immunity.Conclusion Compared with conventional methods,FTS could speed up the rehabilitation for patients with gastric cancer postoperatively without increasing the adverse reactions.