Objective: To isolate the key domain of a novel polypeptide fragment from NGN-β that functions like intact NGF molecule.Methods: NGF-β had been treated with cyanogen bromide and trypsin skillfully. The peptide fragment with the activity inducing PC12 pheochromocytoma cells differentiation was isolated and purified by Sephadex G-50 gel filtration chromatography , DE-52 celluloseion exchange chromatography and C-18 reversed-phase column HPLC after NGF-β cleavaged by CNBr at 9th met then by trypsin at Arg or Lyscleavaged by CNBr at 9th met then by trypsin at Arg or Lys. Amino acid sequencing of this novel peptide fragment was performed by Automatic Amino Acid Analyser and Amino Acid Sequencer. Results: The functional fragment from cleavaged NGF might induce differentiation of PC12 cells. The fragment was consisting of two linear polypeptides . One of them was 16 peptide, GEFSVCDSVSVWVGDK , and other was 14 peptide, HWNSYCTTTHTFVK, linked by a disulphide bridge corresponding to residues 10～25 and 75～88, respectively, of the amino acid sequence of nerve growth factor, the result of biological activity assay in PC12 cells showed that the optimum concentration of this peptide were 0.001～0.1 μg*L-1. Conclusion: A novel peptide inducing differentiation of PC12 cell line of pheochromocytoma cells was obtained in the study. It′s isolation and purification successfully will underlie synthesis or expression of hyperactive neurotrophic small molecular substance although the relationship between the configuration and functions is not clearly.

Objective To observe the changes of the peripheral blood T helper (Th)17 cells/regulatory T cells (Treg) ratio in patients with anti‐tuberculosis drug‐induced liver injury ,and to explore the relationship between Th17/Treg ratio and liver function . Methods Ninety inpatients with the diagnosis of secondary pulmonary tuberculosis (TB) from Affiliated Hospital of Zunyi Medical College from Feb .2013 to Mar .2015 were included in this study .All these patients had normal base line liver function .After 2 weeks of anti‐TB chemotherapy ,21 patients with alanine transaminase (ALT) and/or aspartate aminotransferase (AST) ≥2 × upper limit of normal (ULN) were defined as TB with liver injure group ,the other 69 patients with ALT and/or AST<2 × ULN were defined as TB with non‐liver injured group .Before treatment and two weeks after anti‐TB therapy ,the peripheral blood Th17/Treg ratios and liver functions of all patients in both groups were detected in order to compare the changes of Th17/Treg ratios pre and post anti‐TB chemotherapy and to explore the relationship between Th17/Treg ratio and ALT/AST levels .Student′s t test was used for statistical analysis ,and Pearson correlation analysis was used for correlation test .Results Before treatment ,there was no statistical significance of Th17 ,Treg and Th17/Treg ratio between groups (P> 0 .05) .The Th17 levels in TB with liver injured group ([2 .522 ± 0 .388]/mL vs [2 .075 ± 0 .369]/mL ,t=3 .633 ,P<0 .01) and TB with non‐liver injured group ([2 .326 ± 0 .348]/mL vs [1 .929 ± 0 .402]/mL ,t= 6 .468 , P< 0 .01) ,and Th17/Treg ratios in both groups(0.618±0.104vs0.489±0.107,t=3.553,P<0.01 ;0.554±0.108vs0.450±0.098,t=6 .353 ,P<0 .01) were all increased after anti‐TB chemotherapy ,with statistically significant differences . However ,there was no statistical significance of Treg levels pre and post anti‐treatment in both groups (both P>0 .05) .The Th17 level and Th17/Treg ratio in TB with liver injured group were significantly higher than those in TB with non‐liver injured group (t= 2 .203 , P< 0 .05 and t= 2 .345 , P< 0 .05 , respectively) .In liver injured group ,the Th17/Treg ratio was positively correlated with ALT level (r=0 .849 ,P=0 .044) ,but not with AST level (P>0 .05) .Conclusion The changes of Th17/Treg ratio are related with anti‐TB drug‐induced liver injury in patients with secondary pulmonary tuberculosis .

Objective The aim of the present study is to evaluate the reliability and validity of the Mandarin version of the PP (MPP) .Methods The first step in the establishment was to translate the original English version into mandarin version with the method of cross -culture translation .The reliability was performed with the internal consistency analysis and test -retest reliability .The validity was performed for the content validity and structure va-lidity .The samples were from 80 Chinese CI children ,and 43 parents answered this questionnaire again 1 month lat-er to evaluate the test -retest reliability .The average age at cochlear implantation were 26 ± 14 months ,ranging from 7 months to 68 months ,the average duration of CI use were 10 ± 7 months ,ranging from 0 month to 24 months .Results The reliability analysis indicates that the Cronbach'sαcoefficient was 0 .797 ,except for the well-being and happiness ,education ,whose coefficients are respectively 0 .303 ,and 0 .341 ,all of the other sundomainscoefficient were greater than 0 .5 ,indicating the internal consistency was good .Test -retest reliability of the scale Cronbach'sαwas satisfactory .All subdomains and total score of the scale coefficients were greater than 0 .70(P<0 .01) .The validity analysis indicated that the pearson correlation coefficients among the total scale and the 8 subdo-mains were 0 .395~0 .992 ,the correlation coefficients among each subdomains were 0 .09~0 .654 ,which confirmed with the psychological characteristics ,proving its good structure validity .Conclusion The Chinese version of the PP show s good reliability and validity and can be used to evaluation the quality of life in mandarin CI children.

Objective To analyze membrane attack complex (MAC) expression in different types of idiopathic inflammtory myopathy (IIM).Methods We enrolled 57 cases of dermatomyositis (DM) , 37 cases of polymyositis (PM) ,9 cases of sporadic inclusion body myositis (sIBM) and 15 cases of autoimmune necrotizing myopathy with anti-signal recognition particle antibodies (SRP-ANM) in Department of Neurology at Peking University First Hospital from 2011 to 2014, and used x2 test or Fisher exact test to analyze MAC expression in muscle fibers and endomysial capillaries respectively.Results The total MAC expression in DM, PM, sIBM and SRP-ANM was 75.4％ (43/57), 86.5％ (32/37), 4/9 and 13/15 respectively.The MAC expression in muscle fibers was 50.9％ (29/57), 81.1％ (30/37) , 3/9 and 13/15 respectively.The MAC expression in endomysial capillaries was 49.1％ (28/57) , 24.3％ (9/37) , 1/9 and 6/15 respectively.The MAC expression in muscle fibers and endomysial capillaries of sIBM was less than other types of IIM.The MAC expression in muscle fibers of PM and SRP-ANM was higher than DM and sIBM, but there was no statistically significant difference between PM and SRP-ANM.The MAC expression in endomysial capillaries of DM and SRP-ANM was higher than PM and sIBM, while there was no statistically significant difference between DM and SRP-ANM (x2 =0.397, P =0.574).The MAC expression in four types of IIM had regional distribution, of which 11.6％ (5/43) of DM showed bundle distribution.Conclusion There are differences in the damage of MAC in various types of IIM, the damage of MAC in SRP-ANM indicated the pattern of both DM and PM.

Objective To investigate the effect of α-galactosidase A (GLA) gene mutation on cell autophagy and to elucidate its mechanism preliminarily.Methods Two families were diagnosed by ultrastructural pathological examination,GLA gene activity test and GLA gene mutation screening.Mutant type recombinant expression plasmid of two pedigrees (pcDNA3.1-GFP-ex1 (EX1 group),pcDNA3.1-GFP-ex3 (EX3 group)) and wild type recombinant expression plasmid of GLA (pcDNA3.1-GFP-GLA,GLA group) were constructed.Hela cell line (control group) was transiently transfected with recombinant expression plasmid according to lipofectin transfection.The relative gene expression of Beclin-1 was measured with real-time PCR,and protein expression level of LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and P62/SQSTM1 was examined by Western blotting.Results The LC3 protein values of groups EX1,EX3,GLA and control were 1.495 ± 0.064,1.490 ± 0.020,1.285 ± 0.021,1.260 ± 0.042,respectively;P62/ SQSTM1 values were 0.555 ± 0.086,0.480 ± 0.084,0.785 ± 0.439,0.980 ± 0.278,respectively;Beclin-1 mRNA 2-△Ct values were 0.011 ±0.003,0.008 ±0.002,0.005 ±0.001,0.003 ±0.001,respectively;Beclin-1 protein values were 1.178 ±0.098,1.209 ±0.092,0.931 ±0.100,0.796 ±0.184,respectively.Compared with the wide type group,the level of LC3-Ⅱ/LC3-Ⅰ protein was significantly higher in the mutant type groups(t =5.118,4.984;P =0.007,0.008),though no statistically significant difference was found in the expression levels of P62/SQSTM1 (t =1.052,1.400;P =0.323,0.199).Besides,the expression levels of Beclin-1 mRNA (t =3.800,2.445;P =0.005,0.040) and protein (t =2.424,2.729;P =0.042,0.026) were significantly higher in the mutant type groups.Conclusions GLA gene mutation can induce cell autophagic dysfunction,and signaling pathway of autophagic activation may be Beclin-1 dependent.

To clarify the effect of the sympathetic nerves on the body's physiopathological changes and acupuncture effect by the alphal-AR mediation in the past 30 years.The paper has referred to the database of CNKI and Pubmed,and systematically reviewed the publications in the past 30 years about the research of the alphal-AR mediation of the sympathetic nerves to the body's physiological andpathological changes and acupuncture effect.Alphal-AR not only mediated the sympathetic nerves on the inotropic change of the heart,and the contraction of the vascular smooth muscles,bladder sphincter,and uterine smooth muscles and other physiological effect,but also mediated cardiac arrhythmia,myocardial hypertrophy and other pathological process.In addition,Alphal-AR also mediated the acupuncture signal transmission,and acupuncture was able to adjust the sympathetic nervous tension.The body's physiopathological changes are closely related to alpha1-AR and subtype alteration.The in-depth study of alphal-AR helps to explain physiopathological mechanism of the body,as well as provide theoretical basis and the corresponding pharmacological models for better selective drugs.Studying Alpha1-AR on meridians contributes to the discovery of meridian essence and the substantial basis of acupuncture effects.

Objective To summarize clinical phenotypes and pathological characteristics in myopathies with tubular aggregates (TAs).Methods We reviewed 5 697 patients who performed muscle biopsies in our department between January 2001 and July 2015.We collected the cases with TAs and made classification based on their clinical diagnoses and pathological changes.Results Fifty-seven patients (1.00％) showed TAs in muscle specimens,including 50 (87.72％) males and 7 (12.28％) females.According to clinical,neurophysiological,pathological and genetic analysis,the diagnoses included 23 (40.35％) cases of periodic paralysis,7 (12.28％) cases of chronic alcohol intoxication,6 (10.53％) cases of congenital myasthenic syndrome,5 (8.77％) cases of exercise-induced cramps,3 (5.26％) cases of necrotizing myopathy,1 (1.75％) case of stromal interaction molecule 1-associated myopathy,limbgirdle muscular dystrophy 2E,myotonic dystrophy,myotonia congenita,paramyotonia congenitia,hypothyroid myopathy respectively.Other cases of unknown cause included unclassified distal myopathy,external ophthalmoplegia,white matter lesions,mental retardation,stroke,early onset weakness,pulmonary arterial hypertension.Besides TAs,pathological changes also included necrosis of muscle fibers (3 cases,5.26％),neurogenic changes (3 cases,5.26％) and muscular dystrophic changes (1 case,1.75％).Conclusions Our results indicated that TAs are usually found in males and could present in many types of hereditary or acquired neuromuscular disease as independent or accompanying changes.Periodic paralysis,chronic alcohol intoxication and congenital myasthenic syndrome are 3 major diseases causing myopathies with TAs.

Objective To investigate and analyze oral health status of inpatients with schizophrenia.Method We selected 105 patients from Beijing Anding hospital included hospitalization length 1-5 years group (39 cases) and hospitalization length over 5 years (55 cases) were investigated with Self-made questionnaire and assessment guide for psychiatric care (OAG-PC).Results Tooth loss and oral health status of schizophrenia inpatients were both poor.There was significant difference in the number of people with tooth loss (x2 =7.68,P ＜ 0.05) and mean number of tooth loss (F =9.20,P ＜ 0.05) between two groups.The OAG-PC total score (F=11.1,P＜ 0.05),odour from the mouth (F=4.34,P＜ 0.05),saliva (F=3.93,P＜ 0.05),mucous membranes (F=5.64,P＜ 0.05),teeth or dentures (F=5.34,P＜ 0.05),calculus on teeth (F=15.34,P ＜ 0.05),looseness of teeth (F =14.14,P ＜ 0.05),and No.of teeth in the lower jaw (F =7.70,P ＜ 0.05) in the schizophrenia inpatients with hospitalization length 1-5 years group was better than those with hospitalization length over 5 years,there were significant difference between two groups.There were no significant difference in lips (F =0.69,P ＞ 0.05),tongue (F =0.65,P ＞ 0.05),gums (F =1.87,P ＞ 0.05),appearance of teeth (F=1.52,P＞ 0.05),and No.of teeth in the upper jaw (F=3.58,P ＞ 0.05) between two groups.Conclusions Patients with schizophrenia are special vulnerable people.The more course of the disease was longer,the more the oral health status was worse.Tooth loss,calculus,and looseness of teeth are much more serious and will influence chew function seriously.Society,medical institutions,and families should pay more attention to the oral health status of patients with schizophrenia.And taking the preventative measures against poor oral health can improve quality of life of patients with schizophrenia.

Objective To clone and express Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10 ( EC) and to evaluate the biological activity of the fusion antigen EC in inducing specific cytokines secretion from THP -1 cells. Methods The fusion antigen EC gene was cloned into pET-30 a prokaryotic expression vector and expressed highly in E.coli BL21.Then, the THP-1 cells were stimulated with purified fusion antigen EC of different concentrations (10 and 20 μg/ml).Culture supernatants were collected after 12 h and 24 h, respectively.The secretion levels of IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-αand IFN-γin THP-1 cell culture supernatants were detected using Bio-PlexProTM Assays kit.Results The M.tuberculosis fusion antigen EC was cloned and expressed successfully .The secretion levels of IL-6, IL-8 and TNF-αin EC infected THP-1 cells were significantly higher than those in THP-1 cells (P<0.05).The secretion levels of other cytokines did not change significantly .Conclusion The obtained M.tuberculosis fusion antigen EC has biological activity in inducing the THP-1 cells to secrete a higher level of IL-6,IL-8 and TNF-α.

OBJECTIVETo investigate the effect of RNA interference of IgG gene on the radiosensitivity of the human prostate cancer PC3 cell line.

METHODSPC3 cells were trasnfected via lipofectamine by the shRNA vector FCGR1AshRNA targeting the Fc segment of IgG, using NCshRNA as the negative control. Q-PCR and Western blotting were used to analyze the expression of IgG in the trasnfected cells. The cells were then exposed to ⁶⁰Co γ ray at 0, 2, 4, 6, 8, 10 Gy, and the cell proliferation was evaluated by MTS and the cells apoptosis estimated by flow cytometry at 12, 24 and 48 h.

RESULTSMTS assay showed that ⁶⁰Co γ ray significantly inhibited the proliferation of PC3 cells transfected with FCGR1AshRNA as compared with NCshRNA-transfected and blank control cells (P<0.05). Flow cytometry showed that the cell apoptosis rate was significantly higher in FCGR1AshRNA group than in NCshRNA and blank control groups at 48 h after γ ray exposure (P<0.05). At 12, 24 and 48 h after 6 Gy radiation, the cells in FCGR1AshRNA group showed a significantly lowered proliferation rate and an increased apoptosis rate (P<0.05).

CONCLUSIONThe shRNA targeting IgG gene can significantly enhance the sensitivity of PC3 cells to radiation. The combination of RNA interference targeting IgG gene with radiotherapy may be more effective in the treatment of prostate cancer.

Apoptosis ; Cell Line, Tumor ; radiation effects ; Cell Proliferation ; Humans ; Immunoglobulin G ; genetics ; Male ; Prostatic Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; Radiation Tolerance ; Transfection