Objective To investigate the value of genotype detection for diagnosis of hepatitis B virus (HBV ) infection . Methods 433 HBV pattients from January 2011 to August 2013 were detected by PCR-reverse dot blot hybridization ;the DNA were assaied by PCR ;the HBeAg were tested by ELISA .Results Of the 432 HBV patients ,the rato of genotypes B (68 .13% ) was significantly higher than that of genotypes BC (5 .77% ) and of genotypes C(26 .10% )(P<0 .05);there were no significiant differ-ence in the copy of the HBV DNA among the various genotype (P>0 .05);HBeAg negative rate of genotypes B (23 .82% ) ,geno-types BC(14 .78% ) ,genotypes C(1 .42% )had statistically significant(P<0 .05);Genes associated with disease severity :the ratio of genotype B for patients with mild-to-moderate hepatitis B was 87 .20% ,the rato of genotype C was 9 .34% and genotype BC was 3 .46% ,while the ratio of genotype C was 77 .08% ,genotype BC was 14 .58% ,genotype B was 8 .33% in severe hepatitis B .Con-clusion The genotype of HBV is related to disease severity and the negative rate of HBeAg ,it is not associated with HBV DNA of HBV .

Objective To study the effect of VacA on the secretion of THP-1 macrophages as an individual virulence determinant, and the effect of NF-kB on the secretion of THP-1 macrophages. Methods The recombinant plasmid pDsRed-Monomer-Cl/vacA was transfected into macrophages. The cytokine con-tent of TNF-α or IL-1β in the culture medium was tested quantitatively with ELISA kit, respectively. The content of NO or ROS in the culture medium was tested with Griess reagent or DCFH-DA fluorescent probe. The apoptosis rate of macrophages was tested by flow cytometry. The effect of PDTC, an inhibitor of NF-kB, on the secretion and apoptosis of macrophages transfected with the recombinant plasmids, was also studied. The activity of NF-kB was examined in THP-1 cells by electrophoretic mobility gel shift assay(EMSA). Re-suits At 6 h after transfection, the level of TNF-α and IL-1 β in macrophages transfected with the recombi-nant plasmids was significantly higher than that of the control group (P <0.05). At 6 h or 12 h after trans-fection, the level of NO and ROS in macrophages transfected with the recombinant plasmids was significantly higher than that of the control group (P <0.05). At 16 h after transfection, the apoptosis rate of macropha-ges transfected with the recombinant plasmids was significantly higher than that of the control group (P < 0.05). PDTC decreased the production of TNF-α, IL-1 β, NO, ROS and apoptosis rate induced by VacA. VacA was found to trigger NF-kB activation. Conclusion The over-expression of VacA fusion protein can up-regulate secretion and apoptosis of macrophages. Activation of NF-kB is probably involved in the produc-tion of TNF-α, IL-1β, NO, ROS and apoptosis induced by VacA.

Objective To construct an eukaryotic expression plasmid PeDNA3.1 (+)/Lpp20 and to detect its expression in HeLa cells, and to observe the humoral and cellular immune responses in C57BL/6 mice induced by the Helicobacter pylori Lpp20 DNA vaccine injected intramuscularly. Methods The Lpp20 gene was amplified by PCR. PCR product was subcloned into the eukaryotic expression vector pcDNA3.1 (+)/ Lpp20, and the recombinant plasmid was transfected into HeLa cells using Liposome. After verifying that the Lpp20 antigen gene could be expressed in HeLa cells. Six weeks old C57BL/6 mice were immunized with pcDNA3.1 (+)/Lpp20 or pcDNA3.1 (+) or PBS buffer intramuscularly at 2-week interval for four times. ELISA was used for the quantitative detection of the specific IgG antibody in the sera of C57BL/6 mice and the cytokine IFN-γ in mice spleen lymphocyte culture medium after stimulating by Lpp20. The proliferation response of spleen cells was detected by MTT assay. The Lpp20 gene in muscle was identified by PCR. Results The significant specific antibody titers were detected by ELISA in DNA vaccine groups and the highest titer was 1:1024 after 6 weeks. The cytnkine IFN-γ in mice inoculated with pcDNA3.1 (+)/Lpp20 was increased and reached (410.36±56.23) pg/ml. A significant difference was tested between the experiment group and the control group[(25.26±10.85)pg/ml] ,P <0.01. The proliferation response of spleen cells of DNA vaccine group(SI: 2.37±0.22) was significantly higher than those of mice injected with pcDNA3.1 (+) (SI:1.53+0.47) ,P<0.01. Lpp20 gene could exist constantly in musculature cells of mice. Conclusion The eukaryotic expression recombinant pcDNA3.1 (+)/Lpp20 was successfully constructed. Strong humoral and cellular im-munity can be induced by DNA vaccine of pcDNA3.1(+)/Lpp20 in C57BL/6 mice, which might be helpful for further investigation concerning the immunoprotection of DNA vaccine.

Objective To compare eye expression recognition in stable outpatients with schizophrenia with that in normal controls and to explore the relationships between eye expression recognition and social functioning.Methods 107 schizophrenic outpatients and 66 normal controls matched in age,sex and years of education were assessed with Eye Basic Emotion Discrimination Task(EBEDT) and Eye Complex Emotion Discrimination Task(ECEDT).The patients were also assessed with Social Disability Screening Schedule (SDSS).Results The correct numbers were significantly lower for patients to identify basic emotions of eye expressions(13.2±3.8 vs16.0±2.6,P<0.01) and complex emotions of eye expressions(17.9±4.3 vs 20.6±3.5,P<0.01)than those for controls respectively;the correct numbers to identify anger(3.1±1.0 vs.2.1±1.2,P<0.01),fear(1.8±1.0 vs 1.3±1.0,P<0.01) and disgust(1.8±1.1 vs 1.4±1.2,P<0.05)for controls were higher than those for patients significantly.The correct numbers to identify total basic emotions(r=-0.335,P<0.05)and total complex emotions (r=-0.374,P<0.05)in eye expressions showed negatively correlated with the total scores of SDSS in the patients after controlling age and total score of PANSS.Conclusions The ability to recognize basic and complicated emotions in eye expressions in the outpatients with schizophrenia is lower than that in the controls. It shows positively correlated with social functioning moderately in the patients.

Objective To investigate the effects of ginkgo biloba extract on the early postoperative cognitive function and the serum neuron specific enolase (NSE),S100β,tumor necrosis factor-α (TNF-α),interleukin (IL)-6,superoxide dismutase (SOD) and malondialdehyde (MDA) in patients with meningioma resection.Methods 60 patients with meningioma scheduled for elective intracranial tumor resection were enrolled and randomly divided into two groups (n =30 each):ginkgo biloba extract group (group G) and control group (group C).Ginkgo biloba extract was infused intravenously 30 min before anesthesia induction in group G,while group C received the equal volume of normal saline.Venous blood sample were taken at three time points:30 min before induction (T1),extubation (T2) and 24 h after operation (T3)for determination of serum concentration of NSE,S100β,TNF-α,IL-6,SOD and MDA.Cognitive function was evaluated at 1 d before and 3 d after surgery using Mini-mental State Examination (MMSE),Wechsler Adult Intelligence Scale (WAIS) and Wechsler Memory Scale (WMS).Results Compared with T1,the concentration of NSE,S100β,TNF-α,IL-6 and MDA in serum were significantly increased at T2 and T3 in the two groups,while SOD was decreased (P ＜ 0.05).Compared with group C,the concentration of NSE,S100β,TNF-α,IL-6 and MDA were significantly decreased at T2 and T3 in group G,while SOD was increased (P ＜ 0.05).Compared with 1 d before surgery,the scores of MMSE,digit accumulation,digit breadth (forward),vocabulary association and digit symbol replacement were significantly decreased and tracing connection time was significantly increased on the 3rd day after surgery in the two groups.The scores of digital breadth (reverse) and visual regeneration were significantly decreased in group C (P ＜ 0.05).Compared with group C,the scores of digit accumulation,digit breadth (forward and reverse) and visual regeneration were significantly increased on the 3rd day after surgery in group G (P ＜ 0.05).The incidence of postoperative cognitive dysfunction (POCD) at 3 d after surgery in group G was significantly lower than group C (P ＜ 0.05).Conclusions The pretreatment of ginkgo biloba extract can decrease the incidence of early POCD in patients with meningioma resection,and its mechanism may be related to the reduction of inflammatory reaction and antioxidation.

Objective To study the value of new 16S rRNA gene chip in pathogen detection of neonatal sepsis.Method Newborns with suspected sepsis hospitalized in Peking University Shenzhen Hospital from January 2015 to December 2017 were chosen as the subjects.Blood culture and gene chip were both used to detect the pathogens of these infants.The positive rate,the detection time,and the blood volume needed for detection were compared between the two methods.Result A total of 306 cases of suspected neonatal sepsis were included in the study.Among them,34 (11.1％) were positive for blood culture and 54 (17.6％) were positive for gene chip.98 cases were diagnosed as neonatal sepsis,34 (34.7％) were positive for blood culture,and 52 (53.1％) were positive for gene chip.The positive rate of gene chip was higher than that of blood culture (P ＜ 0.05).For the 5 common pathogens of neonatal sepsis,the positive rate of gene chip was higher than that of blood culture.Time to positivity (TTP) and pathogen identification time of blood culture were (14.6 ± 5.5) h and (72.9 ± 19.0) h,respectively.TTP and pathogen identification time of gene chip were both 3 h.The detection time of gene chip was significantly less than that of blood culture (P ＜ 0.001).The blood volume needed for detection of blood culture and gene chip was 1 ～ 2 ml and 0.5 ml.Gene chip needs less blood volume than blood culture.Conclusion Compared with the traditional blood culture,gene chip can quickly detect the pathogens in the blood with higher positive rate and less blood volume.Gene chip is of great value in the diagnosis of neonatal sepsis.

Objective To investigate the value of serum IgG4 level for the diagnosis of IgG4-related hepatobiliary diseases and the differentiation from other hepatobiliary diseases.Methods A total of 270 patients with hepatobiliary diseases in the People's Hospital of Hunan Province from August 2015 to April 2017 were enrolled in this study,and 20 healthy subjects were selected as controls.The 270patients were divided into eight groups:liver cirrhosis group (n =17),acute pancreatitis group (n =52),chronic pancreatitis group (n =33),cholecystitis and gallstone group (n =27),bile duct carcinoma group (n =30),cholangitis and biliary calculi group (n =41),pancreatic cancer group (n =47),IgG4-related hepatobiliary disease group (n =23).The levels of serum IgG4 were measured by rate nephelometery assay.The sensitivity and specificity of IgG4 levels for distinguishing IgG4-associated hepatobiliary diseases were evaluated by receiver operating characteristic curve.Results The levels of IgG4 of the cirrhosis group and the IgG4 related hepatobiliary disease group were significantly higher than those of the control group (P ＜ 0.05).The IgG4 level in the hepatobiliary disease group was significantly higher than those of the other seven groups (Z =-5.267,-6.802,-5.921,-6.005,-6.173,-6.513,-6.014,P all ＜ 0.01).The area under curve (AUC) for IgG4 level in distinguishing IgG4 associated hepatobiliary diseases and other hepatobiliary diseases was 0.982.When 4.13 g/L was used as the cut off value of diagnosis,the sensitivity and specificity of IgG4for diagnosis were 95.7％ and 96.0％ respectively.The IgG4 levels in twelve patients with IgG-associated hepatobiliary diseases after 2 months of glucocorticoid therapy were significantly lower than those before glucocorticoid therapy (Z =-2.021,P =0.043).Conclusion The elevated serum IgG4 level may not be specific just for IgG4-related hepatobiliary diseases.The cut off value of 4.13 g/L should be very useful for diagnosing IgG4-related hepatobiliary diseases,differentiating from other hepatobiliary diseases and evaluating the therapeutic effect of glucocorticoid therapy.The further detailed verification for these findings should be necessary in clinical practice by increasing the sample size.

Objective To observe the levels of interferon gamma-induced protein 10(IP-10)in children with acute viral and bacterial infection.Methods There were three groups:acute viral infection group(51 ca-ses),acute bacterial infection group(52 cases),and healthy control group(51 cases).Serum IP-10 levels were detected by enzyme linked immunosorbent assay(ELISA),and serum C reactive protein(CRP)were deter-mined with BNⅡ automatic protein analyzer.Results The level of serum IP-10 and CRP were significantly different in different groups(P<0.05).Compared with control group,the level of IP-10 and CRP were higher in acute viral infection group or in acute bacterial infection group(P<0.05).The level of IP-10 was higher in acute viral infection group than that in acute bacterial infection group(P>0.05).The level of CRP was higher in bacterial infection group than that in acute viral infection group(P<0.05).The IP-10 levels in viral Infec-tion patients were not correlated with the CRP levels.The area under curve(AUC),sensitivity and specificity of IP-10 and CRP were 0.688 and 0.873,35.0% and 79.6%,96.1% and 98.0%.When cut-off value of predic-tive probability was 0.713,sensitivity and specificity were increased to 82.5% and 100.0%.Conclusion Ser-um IP-10,CRP and predictive probability are valuable in the diagnosis of acute pathogen infection in children.

Objective This study aimed to assess the diagnostic value of anti-mutated citrullinated vimentin (MCV) antibodies in rheumatoid arthritis (RA) and its correlation with disease progression, extra-articular manifestations and overlap syndrome. Methods Retrospective Studies. Clinical data of 837 patients in PekingUnionMedicalCollegeHospitalfrom June to August 2017 were collected, including the result of anti-MCV, anti-cyclic citrullinated peptide (anti-CCP) antibodies, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR) and High-sensitivity-C-reactive protein (CRP). According to the 1987 American College of Rheumatology classification criteria for rheumatoid arthritis, there were 323 patients diagnosed with RA, including 59 males and 264 females with the average age of 51 years. According to whether the RA patients have overlap syndrome with other autoimmune disease (AID) or have extra-articular manifestations, 258 cases were categorized into RA group, including 47 males and 211 females with the average age of 50 years; 14 cases were categorized into the group of overlap syndrome, including 1 male and 13 females with the average age of 36 years;51 cases were categorized into the group of extra-articular manifestations, including 11 males and 40 females with the average age of 59 years.According to 2010 rheumatoid arthritis classification criteria for destruction in joints, the radiographic changes were divided into 4 stages. There were 203 casesenrolled in our study, 88 caseswere fitted into early stage group (stage I)including 21 males and 67 females with the average age of 48 years; 115 caseswere fitted into progressive stage group, which compromisedstageⅡ (interim stage), stage Ⅲ (severe stage) and stage Ⅳ(final stage) cases, including 19 males and 96 females with the average age of 53 years. Mann-Whitney U test, x2 test, Receiver operating characteristic (ROC) curves and Spearmancorrelation coefficientwere used in Statistical analysis. Results Ⅰ Amongdiagnosed RA patients, 199 (61.6％) cases were positive for anti-MCV, anti-CCP and RFsimultaneously, 42 (13％) cases were positive for anti-MCV, which was higher than anti-CCP positive (1 cases, 0.3％) or RF positive (7cases, 2.2％). The difference was statistically significant(P<0.001, P<0.001). ⅡROC was calculated and MCV=35.95 U/ml was used as best-fit cut-off value. The AUC for anti-MCV was 0.867, while the sensitivity was 80.5％and specificity was 80.9％.ⅢThe detection levels of anti-MCV (682.8 (106.4-1000.0)), anti-CCP (407 (4.0-1536.0)) and RF (82.8 (21.1-244.9)) in the group of progressive stage were higher than those in the group of early stage (114.5 (28.5-1000.0), 62.5 (5.0-1020.7), 50.1 (6.7-127.1)), which showed a significant difference(P<0.05, P<0.05, P<0.05). The anti-MCV, anti-CCP and RF were positively related to the degree of joint destruction (r=0.229, P<0.05;r=0.187, P<0.05;r=0.167, P<0.05);anti-MCV and anti-CCP were positively related to extra-articular manifestation (r=0.152, P<0.05;r=0.136, P<0.05). Conclusion Anti-MCV antibodies are more sensitive in patients with RA, and have complementary diagnostic value for anti-CCP and RF-negative patients; high levels of anti-MCV and anti-CCP in RA patients are associated with RA progression and extra-articular involvement.

OBJECTIVE：To compare the c ontents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products，and to provide reference for the optimization of salt-processing technology. METHODS ：Processed with bittern ， magnesium chloride ，sodium chloride ，calcium chloride and potassium chloride as excipients ，and HPLC method were adopted to determine the contents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products. The effect/toxicity ratio and toxicity component index were used to evaluate the effect and toxicity of raw aconite and 5 kinds of salt-processed products. RESULTS： The linear range of benzoylneoaconitine ， benzoylhypoconitine， benzoylaconitine， neoaconitine， aconitine and hypoconitine were 2.440-24.40，2.240-22.40，2.020-20.20，2.780-27.80，2.240-22.40，2.240-22.40 μg/mL（r≥0.999 0）. RSDs of precision，stability and repeatability tests were all less than 2％. The recovery rates were 102.13％-104.87％（RSD＝0.98％，n＝ 6），100.00％-105.00％（RSD＝2.02％，n＝6），95.00％-99.29％（RSD＝1.77％，n＝6），100.41％-104.58％（RSD＝1.78％，n＝ 6），98.87％-99.90％（RSD＝0.41％，n＝6），100.20％-104.00％（RSD＝1.55，n＝6），respectively. The contents of total alkaloids in order from the largest to the smallest was as follows ：raw aconite ＞processed products of sodium chloride ＞processed products of bittern ＞processed products of potassium chloride ＞processed products of calcium chloride ＞processed products of magnesium chloride. The order of effect/toxicity ratio was processed products of potassium chloride ＞processed products of magnesium chloride＞processed products of sodium chloride ＞processed products of calcium chloride ＞processed products of bittern ＞raw aconite. The order of toxicity component index was raw aconite ＞processed products of sodium chloride ＞processed products of bittern＞processed products of calcium chloride ＞processed products of potassium chloride ＞processed products of magnesium chloride. CONCLUSIONS ：Using 5 kinds of salt as excipients ，the proce ssing technology has different degrees of “efficacy enhancing and toxicity reducing ”effect. Among them ，magne- sium chloride ，potassium chloride and calcium chloride are better for processing and can be used for processing aconite.