Study of coxsackievirus A6 infection in Mongolian gerbils / 中华实验和临床病毒学杂志

Objective@#To study infection of coxsackievirus A6 (CV-A6) in Mongolian gerbils.@*Methods@#To screen the optimal ages of Mongolian gerbils, five groups with different ages were infected with 1×105 TCID50 dose of CV-A6 XS45 strain by intraperitoneal, and symptom scores of Mongolian gerbils were collected. Then to estimate the dose-effect, three doses of virus were injected to the Mongolian gerbils. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry(IHC) were used to determine virus load and tissues infection in muscle, brain and intestinal tract.@*Results@#Mongolian gerbils infected with 1×105 TCID50 dose CV-A6 consistently exhibited clinical signs, and the morbidity (death) rates of five age groups were up to 100%. There was a positive correlation between the trend of symptom scores changes and ages. The morbidity (death) rates of three doses (1×103 TCID50, 1×104 TCID50, 1×105 TCID50) also were up to 100% in 28 days Mongolian gerbils. The correlation between the trend of symptom scores changes and doses were negative. Virus loads were detected in muscle, brain and intestinal tract of pathogenesis animal. The virus loads of muscle were higher than others. IHC results showed virus infection and cytopathic effects in three tissues.@*Conclusions@#Mongolian gerbils had high susceptibility to CV-A6, and were best for animal model of CV-A6 infection.

Effect of Andrographis paniculata Nees extract on coxsackievirus A16 in vitro / 中华临床感染病杂志

Objective To investigate the effect of Andrographis paniculata Nees(APN)extract on Coxsackievirus A16(CVA16)in vitro.Methods African green monkey kidney-derived Vero cells(Vero cells)were treated with APN extract at the concentration of 500.0,250.0,125.0,60.0,30.0,15.0,7.5 and 3.8 μg/mL,the cytotoxicity was determined with cell counting Kit-8 and the IC50was calculated by Probit unit regression method.Direct inactivating activity on CVA16,blocking of CVA16 adsorbing Vero cells and inhibition of CVA16 replication in Vero cells were determined and compared between Ribavirin(RBV) and APN extract with CVA16-infected Vero cells.SPSS 24.0 software was used to analyze the data.Results The selected concentrations of APN extract and RBV for experiment were 15.0, 7.5 and 3.8 μg/mL according to cytotoxicity test.Both of APN extract and RBV had neither direct inactivation on CVA 16 nor blocking of CVA16 adsorbing at the concentration of 15.0,7.5 and 3.8 μg/mL(F=1.54,1.52 and 0.67, 1.68,all P>0.05).However,both drugs had the capability of inhibiting CVA 16 replication in Vero cells at the concentration of 15.0 and 7.5 μg/mL(t=6.87,11.76 and 7.71,12.84,all P<0.05).Conclusion Experimental result shows that APN extract can effectively inhibit CVA 16 replication in Vero cells in vitro.

Effects of CHOICE management mode on adherence of adolescents with type 1 diabetes mellitus / 中国实用护理杂志

Objective To evaluate the effects of CHOICE management mode on adherence of adolescents with type 1 diabetes mellitus (T1DM). Methods A total of 49 adolescents with T1DM were elected using convenience sampling method, and divided into the experimental group (26 cases) and the control group (23 cases) by random number table. The experimental group received the CHOICE management for 1 year, while the control group received traditional education management. The patients′self-management behaviors and HbA1c were evaluated before the intervention and 1, 3, 6 and 12 months after the intervention. Results The scores of Control Problems, Attitude Toward Diabetes and Understanding Management Knowledge after 12-months intervention were (2.71 ± 0.43), (3.89 ± 0.60), (3.05 ± 0.71)points in the experimental group, (2.49 ± 0.33),(4.20 ± 0.40),(3.51 ± 0.70)points in the control group, there were significant differences (t=-2.101, 2.129, 2.289, P<0.05). The scores of Diet Adherence after 1 month and 3 months intervention were (2.85 ± 0.0.67), (2.79 ± 0.66)points in the experimental group, (2.39 ± 0.70), (2.38 ± 0.67)points in the control group, there were significant differences (t=2.170, 2.338, P<0.05). HbA1c was(8.29 ± 0.76)％ in the experimental group, (8.85 ± 1.14)％ in the control group, there was significant difference (t=-2.041, P<0.05). There were no significant differences at the others times between two groups (t=- 1.556-- 0.527, P>0.05). Conclusions CHOICE management mode can effectively improve the self-management behavior of adolescents with T1DM.

Establishment of an animal model for coxsackievirus A16 infection related immunological evaluation / 中国比较医学杂志

Objective To establish a simple and reliable experimental rodent model sensitive to coxsackievirus A16 ( CVA16) .Methods Mongolian gerbils with different age were selected and inoculated intraperitoneally with live CVA16, and the gerbils were observed daily until 14 days postinoculation to screen for the most optimal ages sensitive to the virus.The dose-dependent symptoms were evaluated and the 50% lethal dose (LD50) was determined.The virus titers were measured in blood and various tissues of CVA 16-infected Mongolian gerbils 3 days post-infecton.Finally, the gerbils were immunized twice with inactivated CVA 16 vaccine at day 1 and day 11, respectively, followed by challenge with the virus with a dose of LD50 at day 14.The gerbils were then observed for another 2 weeks to record their body weight , symptom and mortality rate .Their blood samples were collected from the eyes , and CVA16-specific neutralizing antibodytiters and total antibody titers was checked by microneutralization test and ELISA , respectively .Results Various clinical symptoms, such as inactivity, hind limb weakness, paralysis and even death occurred in gerbils following CAV 16 infection. 7-day-old and 14-day-old gerbils are susceptible to CVA 16 infection whereas 28-day-old gerbils are resistant .The most sensitive and appropriate age is 14-day-old.The 50%lethal dose was determined to be 1×104.5 CCID50.High titers of the virus were confirmed in blood and various tissues of Mongolian gerbils contracted CAV 163 days post-infecton.The survival rate is 87.5%for 14-day-old gerbils preimmunized with two doses of inactivated CVA 16 vaccine and challenged with the virus.The geometric mean titers ( GMTs) of neutralizing antibody was 28.14, and the seroprevalence was 87.5%.Conclusions Mongolian gerbils is sensitive to CVA16 and the virus reproduces actively in Vivo.Thus, it can be used as a reliable small animal model for studies of CVA 16 pathogenesis , vaccine development and drug evaluation .

Preparation and identification of a novel anti-Enterovirus 71 vaccine of recombinant fusion protein virus-like particle / 中华临床感染病杂志

Objective To develop a novel anti-Enterovirus 71 (EV71) vaccine as recombinant virus-like particles.Methods By utilizing the foreign antigen presentation and virus-like particles forming features of Norovirus casipid VP1 P domain (NoVP), two pET-28a (+)-based recombinant expression plasmids containing either NoVP alone or NoVP with three specific epitopes SP55, SP70 and VP2-28 of EV71 capsid proteins tandemly inserted at the surface loop site were constructed and transferred to Escherichia coli.The recombinant fusion proteins of NoVP + EV71-SP55-SP70-VP2-28 and NoVP were induced expression and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and transmission electron microscopy (TEM) observation.BALB/C mice were randomly divided into three groups:group A immunized with the recombinant fusion protein, group B immunized with NoVP and group C injected with 10 mmol/L Tris plus 20 mmol/L NaCl (pH 9.0).Enzymelinked immunosorbent assay (ELISA) was used to test specific antibodies in the serum of the mice, besides, the serums were mixed with the EV71 H3-TY strain and Vero cells, then specific antibody titer was examined by microneutralization test.One way ANOVA and Bonferroni test were used to analyze data.Results Both recombinant fusion protein and NoVP were expressed in Escherichia coli in inclusion bodies form.SDS-PAGE demonstrated that the relative molecular weights of recombinant fusion protein and NoVP protein were approximately 43 × 103 and 36 × 103, respectively;positive protein band of about 43 × 103 (relative molecular mass) was detected in recombinant fusion protein by Western Blot.Virus-like particles derived from the recombinant fusion proteins were observed under TEM.ELISA showed that absorbance 490 (A490) of mice serum added in SP70 peptide was significantly higher than those of group B and C (F =13.860,P ＜0.05).And microneutralization test demonstrated that the serum from group A was able to neutralize EV71 at a geometric mean titer above 1:38.Conclusion A novel virus-like particles vaccine against EV71 with good antigenicity and specificity has been prepared, which is able to induce high titer of neutralizing antibody against EV71 in mice.

Prediction and identification of immunodominant linear B cell epitopes in N9 of avian influenza virus H7N9 / 中华实验和临床病毒学杂志

Objective@#To identify immunodominant linear B cell epitopes in neuraminidase of avian influenza virus H7N9.@*Methods@#By using protean algorithms of bioinformatic software DNAStar, antigenicity, hydrophilicity and surface probability of the H7N9 neuraminidase sequence was analyzed and their corresponding average indexes were calculated. Multiple regions containing potential linear B cell epitopes were predicted. Corresponding peptides were synthesized artificially and used in peptide-ELISA individually to check their reactivity to confirmed H7N9 positive human sera, and H7N9 negative human sera was used as control.@*Results@#Seven potential linear B cell epitopes, namely A to G, were predicted according to their relatively strong antigenicity, hydrophilicity and surface probability. All corresponding synthetic peptides reacted strongly with H7N9 positive sera.@*Conclusions@#Immunodominant linear B cell epitopes in neuraminidase of H7N9 were successfully predicted and confirmed. It will facilitate to clarify molecular basis of the antigen specificity and to make respective antibodies.

Prediction and identification of linear B-cell epitopes in the hemagglutinin of H7N9 avian influenza virus in human / 中华临床感染病杂志

Objective To predict and identify liner B-cell epitopes in the hemagglutinin ( HA) of human-infected avian-origin H7N9 influenza virus and analyze the specificity of H7 subtype.Methods Three serum samples collected at different times from the same patient who was confirmed to be infected with H7N9 influenza virus were provided by Shaoxing People’s Hospital, and one serum sample from healthy person was collected as the control.The extracellular region of HA protein was predicted by TMHMM Sever v.2.0.The potential B-cell epitopes were predicted by DNAStar Lasergene’ s Protean, BcePred and ABCpred tools, and the immunogenicity of the predicted B cell antigen epitopes was assessed by indirect enzyme-linked immunosordent assay ( ELISA ) .H7 subtype specificity was analyzed by comparing HA protein amino acid sequence with H7N9 and H1-H16 subtype influenza virus from Genbank using Clustal X 2.1 software, and Cn3D 4.3.1 software was used to detect the distribution and 3D structure of predicted epitopes on the HA protein of H7N9.Results The potential B-cell epitopes may be located in 172-183, 363-380, 452-472 and 491-506 of extracellular N-terminus of HA protein.ELISA showed that four predicted eptiopes specifically reacted with positive serums from patient.Multi-sequence alignment demonstrated that peptide 172-183 and 363-380 had higher H7 subtype specificity compared with amino acid sequences of other subtypes.Moreover, the predicted linear B-cell epitopes all located on the surface of HA protein according to the 3D structure analysis.Conclusion Four potential B-cell epitopes were identified, in which peptide 172-183 and 363-380 have higher H7 subtype specificity, and may be used in the design of epitope-based vaccines and diagnostics tests.

Prediction and identification of immunodominant B cell linear epitopes in capsid proteins VP1 to VP3 of Coxsackievirus A16 / 中华临床感染病杂志

Objective To identify immunodominant B linear cell epitopes in capsid proteins VP1 to VP3 of Coxsackievirus A16 (CVA16) strain YY157.Methods The protean algorithms of bioinformatic software Lasergene were used to analyze antigenicity, hydrophilicity and surface probability of amino acid sequence of CVA16 capsid proteins VP1 to VP3.Multiple regions containing potential lineal B cell epitopes were predicted and their corresponding average indexes were calculated by BepiPred 1.0 Server.Corresponding peptides were synthesized and examined in peptide-enzyme-linked immunosorbent assay (ELISA) individually to check whether it reacted positively or negatively toward sera from children with confirmed CVA16 infection.Results Totally 21 possible B cell linear epitopes were predicted according to their relatively strong antigenicity, hydrophilicity and surface probability.The corresponding synthetic peptides reacted positively with sera of CVA16-infected children in a varying extent.Conclusion Immunodominant B cell linear epitopes of capsid proteins VP1 to VP3 of CVA16 strain YY157 are successfully predicted and confirmed.

Synthesis and biological evaluation of sophocarpinic acid derivatives as anti-HCV agents

Chronic hepatitis C virus (HCV) infection has become a major public health burden worldwide. Twenty-two sophocarpinic acid or matrine derivatives were synthesized and their anti-HCV activities were evaluated in vitro. The structure-activity analysis revealed that (i) sophocarpinic acids with a D-seco 3-ring structure scaffold were more favorable than matrines with a 4-ring scaffold; (ii) the introduction of an electron-withdrawing group on the phenyl ring in 12-N-benzenesulfonyl Δ (βγ) sophocarpinic acids was beneficial for the antiviral activity against HCV. Among them, compounds 9h and 9j exhibited the most potent inhibitory activities on HCV replication with selectivity indies of 70.3 and 30.9, respectively. Therefore, both were selected as antiviral candidates for further investigation.

Effects of health education on diet and daily behaviors in stegmonth / 现代临床护理

Objective To investigate the effects of health education on diet and behaviors in stegmonth. Methods Three hundred and forty three primiparas were divided into the experiment and control group.The 163 primiparas in the experiment group took the courses in the pregnant women’s school and 180 primiparas in the control group did not.The differences of diet and daily behaviors in stegmonth were compared between the two groups.Results There were statistically significant differences in the rate of diet,health behaviors,breast feeding between the two groups(P<0.01).Conclusion The health education for the pregnant woman can enhance their sense of self care,change their wrong behaviors,and improve the rate of breast feeding.

Construction and application of pharmacophore model of benzoylurea derivatives as beta-tubulin inhibitors / 药学学报

Ten pharmacophore models of beta-tubulin inhibitors were established from the training set of seventeen beta-tubulin inhibitors (two categories) with comformer analysis by using the Catalyst software. The optimal pharmacophore model with two hydrophobic units and two hydrogen bond acceptor units were confirmed (RMS = 0.43, Correl = 0.98, Weight = 2.06, Config = 15.97). This pharmacophore model is able to predict the activity of known beta-tubulin inhibitors and can be further used to identify structurally diverse compounds with higher activity.

Correlated Factors of Compliance to Alimentary Control in Hyperuricemia Patients / 中国康复理论与实践

@#Objective To investigate the correlated factors of compliance to dietary therapy in hyperuricemia patients.Methods 388 hyperuricemia patients were investigated with the questionnaire developed by ourselves.Results 119 cases(31.6%)complied to the dietary control program,and 257 cases(68.4%)didn't controlled food and drink or controlled disqualification.The compliance to alimentary control of hyperuricemia patients was correlated intimately with family history,occupation,the degree of nosography,heathy education,propaganda of the masses media,family supporting and satisfaction.Conclusion The alimentary control compliance and related knowledge about dietary therapy of hyperuricemia of hyperuricemia patients were poor.The alimentary control compliance was better in patients with more serious illness,more heathy knowledge,more family supporting,more propaganda of the masses media and more satisfaction.