Objective To rational design HCV DNA vaccine candidates and evaluate their specific immunity to HCV in mice. Methods We design to construct two DNA vaccine candidates, one consists of E_2 (the envelope glycoprotein 2 of HCV) gene only, the second consists of E_2-gAD (Globular Domain of Human Adiponectin) fusion gene via overlapping PCR. Confirm the expression of the DNA vaccines by Western blotting, and then vaccinated by injection of DNA vaccines with gene electrotransfer (GET) in BALB/c mice. The immune response was measured by IFN-gamma ELISPOT. Results The DNA vaccine candidate consists of E_2-gAD could effectively express in vitro , and it could induced a higher anti-HCV T cell response in mice than the one consists of E_2 only. Conclusion The HCV DNA vaccine consists of E_2-gAD fusion can increase the immunity of the E_2 to some extend, and the research paved a way to develop and optimize the novel HCV DNA vaccine.

Objective To investigate the expression of neural-nitric oxide synthase (nNOS) in renal clear cell carcinoma and its clinical significance.Methods The expression of nNOS mRNA in 533 samples of TCGA database was analyzed with Student t test,and statistical analysis was performed to assess the relationship between nNOS expression and clinical prognosis with Kapla-Meier test.Western blot analysis of nNOS protein expression in 10 cases of clear cell renal cell carcinoma(ccRCC) from department of urology of Wuhan union hospital with student t test.Results The mRNA levels of nNOS in 72 cases of ccRCC in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.57 ± 0.17,it is significantly lower than those in adjacent tissues (P ＜ 0.01).The mRNA levels of nNOS in 533 cases of ccRCC,in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.76 ± 0.05,it is significantly lower than those in adjacent tissues (P ＜ 0.01).A total of 533 sample studies showed a low correlation between nNOS expression and clinical T stage,T1-1.59 ±0.08,T2-1.96 ±0.13,T3-1.90 ±0.09,T4-2.38 ±0.28 (P =0.0029) and -1.63 ±0.06 and-2.16 ± 0.13 between non-metastasis and no-metastasis (P =0.0009),and-1.57 ± 0.08 and-2.03 ± 0.11 between non-recurrence and recurrence (P =0.008).Survival analysis showed that the overall survival time were (40.3 ± 5.6) months and (48.3 ± 5.7) months in lower and higher nNOS expression,and disease free survival time were (37.1 ± 2.1) months and (40.3 ± 5.6) months in lower and higher nNOS expression,both with shorter time in low expression of nNOS (P ＜ 0.01).nNOS proteins were 1.02 ± 0.16 and 0.61 ± 0.1 1 in tumor tissues and adjacent tissues with significantly lower expression(P＜0.05).Conclusions The mRNA and protein of nNOS are lower in ccRCC with a poor prognosis of ccRCC.

Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene(from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell re-spoase than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fu-sion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.

Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.

Objective To investigate the clinical effect of preoperative neoadjuvant chemotherapy by thalidomide combined with gefitinib for non-small cell lung cancer (NSCLC) patients and provide clinical basis for NSCLC therapy.Methods From 2011/1 to 2013/1,we collected 160 NSCLC cases in our hospital and divided them into 2 groups randomly,80 cases in control group and 80 cases in observation group.The patients in the control group were treated with conventional surgical treatment and patients in the observation group were treated with preoperative neoadjuvant chemotherapy by thalidomide combined with gefitinib before surgery.The clinical effects,the 6 months and 1 year survival rates,and the toxicity effects were observed,and the clinical effects and survival rates between the 2 groups were compared.Results The observation group had a therapeutic efficiency ratio (TER) of 80.0％ which was significantly higher than the control group (50.0％),and the difference was statistically significant (P＜0.05).The 6 months survival rate and 1 year survival rate were 97.5％ and 92.5％,which were both significantly higher than those of control group (87.5％ and 77.5％),and all differences were statistically significant (P＜0.05).The toxicity effects of observation group were lower.Conclusions Preoperative neoadjuvant chemotherapy by thalidomide combined with gefitinib for NSCLC therapy is effective and safe,which is worthy of study and further application in clinical treatment..

OBJECTIVETo probe factors of influencing therapeutic effects of acupuncture in the patient of insomnia.

METHODSAccording to scores of degrees of anxiety and depression, 52 cases of insomnia were divided into 3 groups, group I (mild or less degree) and group II (moderate degree) and group II (serious degree). The Pittsburgh sleep quality index (PSQI) were compared before and after treatment in the 3 groups, and between two groups after treatment. Results There were significant differences in the therapeutic effect as the groups I, II compared with the group III (P < 0.01). The total sleep quality in the group I was better than that in the group II (P < 0.05).

CONCLUSIONThe degree of anxiety and depression in the patient of insomnia is one of important factors influencing therapeutic effect of acupuncture on insomnia.

Acupuncture Therapy ; Anxiety ; Depression ; Depressive Disorder ; Humans ; Sleep Initiation and Maintenance Disorders

OBJECTIVETo evaluate the relationship between children's temperament and dental fear.

METHODS254 children(aged 4-6 years) during first dental treatment took part in the investigation. Their parents answered the Chinese preschool children's temperament scales (CPTS). The Frankl method was used to classify the degree of the children's dental fear. The K independent samples test and One-way ANOVA test were performed to find the differences of the type of temperament and the scores of temperament dimension among three groups.

RESULTSAmong the 254 children(aged 4-6 years), 104 had no fear, 80 had fear and 70 had extreme fear. The incidence of dental fear in children was 59.06%. There were no statistical differences (P > 0.05) of dental fear between boys and girls. There were statistically significant differences for the type of temperament among no fear group, fear group and extreme fear group. The scores of adaptability and quality of mood were higher in the extreme fear group and fear group than that in the no fear group. The differences in scores of adaptability and quality of mood was statistically significant between the extreme fear group and no fear group. But the scores of other seven temperament dimensions had no statistical significant differences among three groups.

CONCLUSIONChildren's dental fear is correlated to their temperaments. The tendencies of negative mood and slow adaptability should be considered that the patients were at risk of developing dental fear problem.

Child ; Child Behavior ; Child, Preschool ; Dental Anxiety ; Female ; Humans ; Male ; Temperament

Hemorrhage ; etiology ; Humans ; Infant ; Male ; Mouth Diseases ; etiology ; Vitamin K Deficiency ; complications ; diagnosis

OBJECTIVETo analyze the characteristics of emergency facial trauma in children and mother's cognition on it.

METHODSDescriptive analysis was undertaken for 216 children with maxillofacial trauma. The analysis included age distributions of the children, reasons and places for the trauma, and positions and severity degrees of the trauma. The questionnaires were carried out for the patients' mothers, to understand the cognition degrees and prognosis factors.

RESULTSMost of the patients with facial trauma were aged 1-6, with the male and female ratio of 1.4:1. Falls were the leading cause for the maxillofacial injuries(83.33%). The most common types of injury were the soft tissue injuries (63.43%) and the teeth injuries (31.94%), the jaw fractures were least. 50.00% accidents occurred at home. 97.22% mothers had no knowledge of the facial injuries in children. 69.44% mothers thought it avoidable. It were relevant between mothers' educations and injuries times (chi2 = 18.16, P < 0.05).

CONCLUSIONMothers lack the knowledge is one of the most important reasons for the maxillofacial injuries in children. Propaganda should be increased among them to reduce risks of maxillofacial injuries.

Child ; Child, Preschool ; Cognition ; Facial Injuries ; Female ; Humans ; Male ; Maxillofacial Injuries ; Parents ; Skull Fractures ; Soft Tissue Injuries ; Tooth Injuries

AIM:To explore the changes and significance of Kupffer cells in the process of tree shrew chroni -cally infected with hepatitis B virus (HBV).METHODS:The animals were divided into 3 groups.Group A consists of 6 tree shrews that were identified as persistently infected with HBV;group B consists of 3 tree shrews that were suspected as persistently infected with HBV;group C consists of 4 tree shrews that were not inoculated with HBV and were applied as normal controls.Liver biopsies were collected regularly from all animals , and the Kupffer cells were isolated , purified and primarily cultured.The techniques of flow cytometry , immunohistochemistry, lysosomal fluorescent probe staining and real-time RT-PCR were applied to determine the number and function of these Kupffer cells .RESULTS: The result showed that the count and proportion of CD 163+cells in group A were significantly higher than those in group B and group C ( P<0.05).Meanwhile, the fluorescence intensity levels of lysosomal , the number of lysozyme-positive cells and the mRNA ex-pression level of TNF-αin the Kupffer cells in group A were significantly lower than those in group B and group C ( P<0.05).CONCLUSION:Kupffer cells may play a regulatory role during host’s chronic HBV infection.