0.05). Short-term memory and logic memory(LM) scores were positively correlated with entorhinal cortex volumes(r=0.484, 0.529; all P

AIM: To investigate the molecular mechanism of Epstein-Barr virus encoded latent membrane protein 1 regulated cellular proliferation in nasopharyngeal epithelial cells. METHODS: The nasopharyngeal epithelial cells NP69 were infected with RV-pLNSX (the empty vector) and RV-LMP1 retroviruses, respectively. Therefore, the NP69-pLNSX and NP69-LMP1 cell lines were established. Sequentially, cellular proliferation of NP69-pLNSX and NP69-LMP1 cells was compared to draw the cellular growth curve. The experiments of plate clone formation and forming of soft agar colony were conducted. Meanwhile, the differential expression of proteins were identified between NP69-pLNSX and NP69-LMP1 cell lines by proteomic methods, and the expression levels of partial identified proteins were verified. RESULTS: (1) LMP1 was able to accelerate cellular proliferation of nasopharyngeal epithelial cell NP69 (n=3, P<0.05). (2) Twenty two proteins (9 up-and 13 down-regulated) of LMP1 mediated regulation were identified from infected NP69 cell lines, and the differential expression of partial identified proteins was confirmed by Western blotting and fluorescent real-time quantitative RT-PCR. CONCLUSION: LMP1 probably mediates the regulation of vimentin protein and keratin 19 protein expression to promote cellular proliferation in NP69 cells.

Background Congenital cataract is a major cause for blindness of childhood.Genetic gene mutation accounts for almost 1/3 of congenital cataract patients.The most common inheritance type is autosomal dominant congenital cataract (ADCC).Over 100 mutations in 26 genes have been found to be associated with ADCC.Objective This study was to identify the disease-causing gene mutation in a family with ADCC.Methods This study was approved by Ethic Committee of Beijing Tongren Hospital and followed Declaration of Helsinki.A northern Chinese family with autosomal dominant congenital nuclear cataract was entrolled in Beijing Tongren Hospital in January 2011.Ocular examinations were performed and periphery blood specimens were collected from each family member under the informed consent.Genomic DNA was extracted.Twenty-one microsatellite markers around 17 ADCC genes were selected for linkage analysis,and two-point LOD score was calculated.CRYGC gene and CRYGD gene were amplified and screened for mutations using direct sequencing.ProtScale software was used to analyze the changes of hydrophobicity of the mutated protein.Co-segregation of the observed change with the disease phenotype was further detected by restriction fragment length polymorphism (RFLP).Results This family included 20 members of 4 generations,and 9 patients were examined in serial 4 passages,which conformed to autosomal dominant inheritance pattern.Clinical examination revealed binocular congenital nuclear cataract in the 9 patients.Maximum two-point LOD score was 4.68 at marker D2S325 (θ=0).A known T→C change at position 127 of cDNA sequence was found by mutations screening of CRYGD gene.ProtScale programs showed an obvious increase of the local hydrophobicity in the mutant protein.RFLP results indicated that this missense mutation co-segregated with affected members of the family,but was absent in unaffected members and 100 unrelated controls.Conclusions c.T127C mutation of CRYGD gene appears to be the molecular pathogenesis of this ADCC family.Aberrant structure of mutant CRYGD protein caused by hydrophobicity change may lead to opacification of lens.

Objective:To optimize the extraction process of total flavonoids from Chloranthus japonicus sieb by Box-Behnken re-sponse surface methodology. Methods:The independent variables were the ethanol concentration ( X1 ) , liquid-solid ratio ( X2 ) and extraction time ( X3 ) , the dependent variable was the extraction amount of total flavonoids ( Y) , and the extraction process was opti-mized by Box-Behnken response surface methodology. Results: The results showed that the optimal extraction conditions for total fla-vonoids from Chloranthus japonicus sieb were as follows: the ethanol concentration was 54. 8%, the liquid-solid ratio was 13. 6, and the extraction time was 2. 0 h. The verified extraction process parameters of three batches showed that the relative error between the ex-perimental values and those predicted from the regression model was -2. 34%. Conclusion:The optimal extraction process parameters are simple and more convenient with higher precision for the extraction of total flavonoids from Chloranthus japonicus sieb.

Objective distributing character and current researching status of literatures on treatment and prevention of influenza with Chinese medicine in 60 years after foundation of RPC were analyzed to provide a reference for Chinese medicine prevention and treatment of influenza. Methods CBMdisc was used as data source, ibliometrics method was adopted to analyze literatures on the treatment and prevention of influenza with Chinese medicine form 1949 to 2009. Results There was altogether 825 literatures form 1949to 2009. From 1958 to 1959, the volume of document come into a slow growth trend, reaching a high point in 1959, with the volume of document is the 45, may be related to 1957-type pandemic in Asia; document in 1970,growth accelerated beginning in 2003, faster growth, posting the largest for 2005, issued a total of 89, issued a document was an increasing trend in 2009. Conclusion Chinese medicine treatment of influenza in the past 60years, won the sustainable development, Chinese medicine treatment of influenza has become the dominantdisease, Chinese medicine treatment of influenza is promising.

Objective To explore the roles of p53 in ionizing radiation induced MCF-7 cell cycle uncoupling. Methods The p53 knock-down models was established in MCF-7 with retrovirus packaged particles from 293T cells through calcium acid phosphate co-precipitation, then Western blot was used to detect the protein expression. Flow cytometry(FCM) was used to analyze the cell cycle uncoupling and polyploid after irradiation. Results Compared with p53+/+ group, the percentages of G0/G1 cells in p53 -/- group decreased, while those of S and G2+M increased (P < 0.01). In polyploidy analysis 2N cells decreased, whereas both 4N and 8N cells =increased (P<0.01). Compared with sham-irradiation, 4 Gy X-ray led to the decrease of G0/G1, S cells, and the increase of G2+M cells. The increase of 2N cells and decrease of 4N and 8N cells were observed in both p53+/+ and p53-/- cells. Compared with p53+/+ +IR group, the decrease of G0/G1 and S cells and the increase of C2 + M cells were significant (P < 0.01) in p53-/-+ IR groups. 2N cells decreased, 4N cells increased, but no changes in 8 N cells occurred. Conclusion Radiation might induce G2 arrest and cycle uncoupling, p53 plays a role in the regulation of G2 arrest, but no role in cycle uncoupling.

Objective To evaluate the importance of plasma MicroRNAs in early diagnosis of acute myocardial infarction (AMI).Methods 24 patients with AMI as the test group and 20healthy volunteers as the control group were enrolled in this study.Plasma levels of microRNA-1,microRNA-133a,microRNA-208a and microRNA-499 were detected by quantitative real time polymerase chain reaction (qRT-PCR) at 3 h,6 h,12 h,24 h,48 h,72 h after the onset of AMI.Results Plasma microRNA-1 level was greatly increased and reached the peak at 3 h after AMI,then was decreased gradually to normal level at 72 h after AMI.Plasma microRNA-133a level was significantly elevated at 6 h after AMI,reached peak at 12 h after AMI,then was decreased to normal level at 48 h after AMI.Plasma microRNA-1 and microRNA-133a levels were correlated with cTnI expression.The peak time of microRNA-1 was earlier than that of cTnI,while the peak time of microRNA-133a was the same as that of cTnI.Conclusions Increased circulating microRNA-1 and microRNA-133a may serve as potential and novel biomarkers for early diagnosis of AMI.

OBJECTIVE To study the roles of copper transporter 1 (CTR1 )and Cu2 + transporting ATPase αpolypeptide (ATP7A)in lead exposure-induced copper accu mulation and oxidative stress in rat C6 glio ma cells.METHODS Methyl thiazolyl tetrazoliu m (MTT)assay was performed to determine the proper Pb doses (without affecting cell viability)by treated the cells with lead acetate 0 -100 μmol·L -1 for 24 h and 48 h.Superoxide dis mutase (SOD)activity or malondialdehyde(MDA)level were detected respectively by xanthine oxidase technique and thiobarbituric acid (TBA) method.Ato mic absorption spectrophoto metry was e mployed to determine the intracellular levels of Pb and Cu ions.Real-ti me quan-titative PCR and Western blot were used to detect the mRNA and protein levels of CTR1 and ATP7A, respectively.RESULTS The cell viability significantly decreased when the doses of Pb treat ment was higher than 10 μmol·L -1 ,so 10 μmol·L -1 was chosen as a working concentration of Pb exposure in this study.Co mpared with those in the normal controls,a moderately decreased T-SOD activity and an increased MDA level was determined in the cells treated with Pb 10 μmol·L -1 or Cu 5 μmol·L -1 alone, while a significant drop of T-SOD activity and a re markable increase of MDA level was found in the cells co-exposed to Pb and Cu (P<0.01 ).Pb exposure for 24 and 48 h increased the cellular Cu uptake by 1 .2 and 2.5 fold,respectively (P <0.01 ).Evidences fro m RT-PCR showed that Pb exposure for 24 and 48 h upregulated the CTR1 mRNA level by 23.2% and 58.7%,and downregulated the ATP7A mRNA level by 58.1 % and 50.0%,respectively.Results fro m Western blot confirmed that Pb exposure also resulted in an increased CTR1 expression and a decreased ATP7A expression at protein level (P<0.01 ).CONCLUSION Pb exposure lead to Cu accu mulation,by affecting the expression levels of CTR1 and ATP7A,and increased oxidative stress in C6 cells.

Objective To assess the characteristic of normal tension glaucoma (NTG) patients with mild cogni?tive impairment (MCI). Methods This study included twenty-six cases of normal tension glaucoma patients who were diagnosed at ophthalmology department of Peking University Third Hospital. All the participants were examined by us?ing the scales of Montreal Cognitive Assessment (MoCA), mini-mental state examination(MMSE), clinical dementia rat?ing (CDR), activities of daily living (ADL), Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD), Pitts?burgh Sleep Quality Index (PSQI) and polysomnography (PSG). The patients were then divided into Group with MCI (16 cases) and Group without MCI (19 cases). Results There was no difference between the two groups in gender, age, edu?cation, depression, anxiety and body mass index (BMI) (P>0.05), but significant difference in MoCA scores (P<0.05). The incidence rate of sleep disorder of PSQI in was 28.6%(10/35 patients) of total NTG patients, 43.8%(7/16 patients) in Group with MCI, and 15.8% (3/19 patients) in Group without MCI, respectively. The PSQI score was higher in Group with MCI than in Group without MCI (P<0.05). Sleep efficiency was higher in Group without MCI than in Group with MCI (P<0.05), but no difference was found between the two groups in six other indexes of PSQI (P>0.05). The in?cidence rate of sleep structure disorder of PSG in all the NTG patients was 85.7%(30/35 patients), 87.5%(14/16 pa?tients) in Group with MCI, and 84.2%(16/19 patients) in Group without MCI. Sleep time of NREM-N3 was significant?ly shorter in Group with MCI than in Group without MCI (P<0.05), but no difference was found between those groups in total sleep time, sleep efficiency, sleep latency and REM time (P>0.05). Conclusion NTG patients with mild cogni?tive impairment are more prone to sleep disorders, especially sleep structure disturbance and short NREM-N3 time may affect cognitive function.

This study was aimed to investigate the effect of Radix Bupleuri extract (RBE) on 5-HT3R channel currents of primarily cultured hippocampal neurons in depression emotion rats.Depression emotion model ratswere duplicated.RBE was used for drug intervention.And then,the rats were evaluated by the open-field test (OFT) and the sucrose preference test.Serum of rats in each group was collected and then added into the primary cultured hippocampal neurons for 24 h.The 5-HT3R channel currents were recorded by the whole-cell patch clamp.The results showed that compared with the normal group,the total score of OFT in the model group was significantly decreased (P < 0.01); the sucrose preference ratio decreased obviously (P < 0.01); and the current density value of primary cultured hippocampal neurons in serum of the model group was significantly higher (P <0.01).Compared with the model group,the total scores of OFT in the RBE group and fluoxetine group increased significantly (P < 0.05,P < 0.01); the sucrose preference ratio also increased obviously (P < 0.05,P < 0.05); and current density value of the primary cultured hippocampal neurons in serum of the RBE group and fluoxetine group decreased significantly (P < 0.01,P < 0.01).It was concluded that RBE can effectively correct the abnormal 5-HT3R channel currents of rats with depression emotion,which may be one of the central mechanisms in the treatment of depression emotion.