Objective To observe the prophylactic effects of a HSV-2gD2DNA vaccine in guinea pigs challenged with HSV-2strains.Methods Female guinea pigs were divided into3groups with10each,which was immunized intramuscularly with100?g of pc-gD plasmids(recombinant HSV-2DNA vac-cine),or with pcDNA3blank plasmids,with normal saline as control,respectively.Two booster injections were given on day7and day21.Sera were collected for virus neutralization test on day0,day28,and day56.The animals were challenged with HSV-2strain sav intravaginally,and lesions induced on the external genital skin were scored between day1and day21after challenge.Results The titer of neutralizing anti-body to HSV-2was much higher in the sera from animals immunized by pc-gD plasmids than that from ani-mals immunized by pcDNA3blank plasmids or normal saline.Furthermore,the lesion scores on external genital skin were significantly decreased in pc-gD group than those in other two groups with either primary or recurrent infections.Conclusion The constructed gD2vaccine can efficiently protect guinea pigs from genital infection and reduce recurrent infection induced by latent herpes simplex virus.

Objective:To assess the periodontal diseases as a possible risk factor for preterm and low birth weight. Methods: A population-based, cross-sectional study was carried out in Hubei. The samples were chosen randomly by the rule of admittance. Socio-demographic information relating to health and maternal habits was collected through a questionnaire. The clinical parameters were debris index-simplified (DI-S), sulcus bleeding index (SBI), probing pocket depth (PD) and clinical attachment loss (CAJ). The birth time and weight were recorded. The result was analyzed with chi-square test and logistic regression analysis.Results: The incidence of preterm low birth weight was 12.3%. The gingivitis and periodontitis were diagnosed in 86.6% and 64.2% of preterm low birth weight cases. The odds ratio of having gingivitis and preterm low birth weight was 1.30(95%confidence interval: 0.53-3.22). The odds ratio of having periodontitis and preterm low birth weight was 2.69(95%confidence interval: 1.37-5.27). Conclusion: The periodontal diseases of pregnancy can be regarded as an important risk factor for preterm low birth weight.

Objective To compare the therapeutic effect of conventional treatment and conventional treatment plus external application of Shuangbai Powder for patients with wounded limb injured by venomous snake. Methods One hundred patients bitten by venomous snake were randomized into treatment group and control group, 50 cases in each group. The control group was given conventional treatment including repeatedly washing the wound with hydrogen peroxide, debriding the wound, letting blood and draining toxicity, local blocking with chymotrypsin, wet packing the wounded limb with magnesium sulfate, and injection with anti-venomous serum, tetanus antitoxin, antibiotics, furosemide and energy mixture. The treatment group was given external application of Shuangbai Powder on the basis of the treatment for the control group. Both groups were given the comprehensive nursing of psychological nursing, wound nursing, dietary nursing, defecation nursing and functional nursing. The swelling-subsiding time for the wounded limb and pain scores of visual analogue scale (VAS) in the two groups were compared. Results(1) After treatment, swelling-subsiding time for the wounded limb of the treatment group was shorter than that of the control group, the difference being statistically significant (P < 0.05). (2) After treatment for 4 days and at the end of the treatment, VAS scores of the two groups were decreased compared with those before treatment(P < 0.05), and the decrease of VAS scores in the treatment group after treatment for 4 days was superior to that in the control group (P < 0.05). Conclusion The conventional treatment plus external application of Shuangbai Powder is an effective therapy for patients with wounded limb injured by venomous snake by shortening swelling-subsiding time and relieving pain for the wounded limb .

BACKGROUND: CpG oligodeoxynucleotide (ODN) is a type of highly effective immune adjuvant with low toxicity, which has an extensive application in gene therapy for many diseases. However, the specificity for species and cells leading to low uptake by cells and degradation by nuclease blocks its clinical application. OBJECTIVE: To explore the specific delivery and its immunologic efficacy of CpG ODN targeting B lymphocytes of umbilical cord blood by CD40 ligand-receptor-mediated carrier system. DESIGN, TIME AND SETTING: An observation and control experiment was performed at the Department of Hematology, and Department of Pediatric, the First Affiliated Hospital of Jinan University from April 2004 to October 2007. MATERIALS: Fresh umbilical cord blood with heparin was obtained from healthy, natal infant. Informed consent was obtained from his parents, and the experiment was approved by the hospital Ethics Committee. METHODS: CD40 ligand (CD40L)-EDC-PLL-CpG ODN conjugated complex was prepared. Mononuclear cells (MNCs) from umbilical cord blood were co-cultured with conjugated complexes. Uptake rate, mean fluorescence intensity of FAM marked CpG ODN, expressions of MNCs, proliferations of lymphocytes and the IgG levels of culture supematants were detected by flow cytometry, fluorescence techniques, MTT assay and ELISA, respectively. MAIN OUTCOME MEASURES: The uptake rate, the mean fluorescence intensity of CpG ODN by MNCs, subgroups and proliferations of lymphocytes, and IgG levels of culture supematants. RESULTS: Compared to the pure CpG ODN group, the uptake rate of the conjugated complexes group was higher (98%), the peak level of up-taking occurred earlier, and intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, and CD20+ was increased, A value and IgG levels in supematants were all higher than that of the control group. CONCLUSION: CD40 tigand-receptor-mediated carrier system is helpful for CpG ODN delivery targeting to B lymphocyte, enhancing its immunological efficiency.

AIM: To design,prepare and screen out functional small interfering RNA(siRNA) for specifically silencing proliferating cell nuclear antigen(PCNA) gene expression and effectively inhibiting cell proliferation on human hepatoma cell line SMMC-7721,human gastric carcinoma cell line SGC-7901 and human colorectal carcinoma cell line Caco2.METHODS: PCNA siRNA was designed based on previous studies about design guidelines and synthesized in vitro by T7 Mega short script reaction kit according to the manufacturer's instructions.After purification,the integrities of siRNA were identified through 19% denaturing polyacrylamide gel electrophoresis,and the concentrations of the generated siRNA were measured by testing the absorbance at 260 nm using a spectrophotometer.Four synthesized double-strand siRNA were transfected into three types of carcinoma cell lines by LipofectamineTM2000 reagent,respectively.WST-8 assay was employed to examine the proliferative inhibitions of these three cell lines.The subsequent alterations on PCNA mRNA and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochemistry,respectively.RESULTS: 3 sequences,No.2,No.3 and No.4 PCNA siRNA showed effective inhibitions on tumor cells among the 4 candidate siRNA,and a single dose of 50 nmol/L of No.2,No.4 PCNA siRNA showed the most effective inhibitory rates as more than 62% at 48 h after transfection.50 nmol/L of No.2 and No.4 PCNA siRNA transfection caused 72% decrease of PCNA mRNA level and almost completely loss of the protein in human Caco2 cells.CONCLUSION: No.2 and No.4 PCNA siRNA have been screened out in this study,which show the capability to effectively down-regulated PCNA expression and significantly inhibit the proliferation of carcinoma cells.The optimal concentration is 50 nmol/L and satisfactory effects are achieved 48 h after transfection in vitro.

98%), the peak level of uptake occurred earlier, intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, CD20+ increased significantly. A_~570 values of MNCs proliferation and IgG levels in supernatant were all higher. CONCLUSION: CD40 ligand-PLL carrier system may delivery CpG ODN targeting to B lymphocytes, enhancing its immunological efficiency.

AIM: To evaluate the effect and safety of Rhodamine 123 (Rh123)-mediated photodynamic treatment (PDT) on acute graft versus host disease. METHODS: An acute graft versus host disease (aGVHD) mice model was established using C57B/6 mice as donors and BALB/c mice as recipients. Mixed lymphocytic cells were cultured with Rh123 (50 nmol/L) and irradiated by argon laser 30 mW/cm2 for 3 min, then transplanted to BALB/c recipient mice mixed with donor bone marrow. Hepatopoietic recovery, aGVHD occurrence, survival time after transplantation and pathological changes were observed. In addition, CD3+CD69+ positive rates of MLC were examined by flowcytometry. RESULTS: Occurrence of aGVHD decreased, degree of pathological manifestation became milder, survival rates were higher than non PDT groups. CD3+CD69+ rates of MLC cells treated with photodynamic therapy (PDT) and cultured for 24 h significantly decreased. CONCLUSION: Rh123-mediated PDT can effectively prevent aGVHD of allogeneic bone marrow transplantation in mice.

Objective To study the effects of community-based rehabilitation on cerebral apoplexy patients with cognitive disorders.Methods 30 cases of cerebral apoplexy patients were enrolled according to relevant standards,who were given standard rehabihtation care in Community Health Service Center(training group) ,while another 30 similar cases from subordinate health service stations (control group) given only rehabilitation guidance once a week.Two groups were not do professional cognitive training.All patients were assessed with Mini Mental State Examination( MMSE), Fugl-Meyer Assessment(FMA) and modified Bathel index rating scale(MBI) to evaluate their cognition ,movement function and activity of daily living(ADL) before training as well as after 4-week and 8-week training.Results There was no signficant difference in cognition,movement functions and ADL between training group and control group before treatment(P>0.05).The differences in the assessment of Brounnstrom scale and MBI between two groups after 4-week training were not significant( P > 0.05 ).The scores of Fugl-Meyer scale, MMSE and Barthel index of the training group after 8-week training were significantly higher than those of control group( all P < 0.05 ).Conclusion Community-based rehabilitation training could improve the movement functions and ADL,and cognitive functions of cerebral apoplexy patients.

Objective:To study the antitumor mechanism of W40,a monomer purified from Wedelia prostrate (Hook.et Arn.) Hemsl.Methods:The effects of W40 on the cell proliferative of GLC-82 cells were detected by MTT assay and colony formation assay.The migratory abilities of GLC-82 cells were observed by wound healing assay.Cell apoptosis was evaluated by Annexin V-FITC/PI staining analysis.The levels of apoptosis-relative proteins and cell proliferation-related proteins,such as Caspase-3,PARP,Stat3 and ERK,were detected by Western blotting.Results:MTF assay showed that W40 had a significant cytotoxic effect on non-small cell lung cancer GLC-82 cells.Colony formation assays showed that W40 significantly inhibited GLC-82 cells proliferation.The migration of GLC-82 cells was inhibited by W40 in a dose-dependent manner.Flow cytometry showed that the apoptotic rate increased gradually in a concentration-dependent manner.W40 down-regulated Stat3 as decreasing p-Stat3 and downstream proteins of Bcl-2 and Mcl-1.At the same time,W40 up-regulated the expression of pro-apoptotic protein Bax,and increased the cleavaged Caspase-9,Caspase-3 and PARP.W40 also down-regulated BRAF / MAPK / ERK signal pathway as decreasing p-BRAF,p-MEK and p-ERK.Conclusions:W40 induced apoptosis by inhibiting BRAF / MAPK / ERK and Stat3 signaling pathways.

Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein expression of Nrf2/Bach1 in human skin fibroblasts (HSFs).Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0,2.5,5,10,20 and 40 mg/L for 24 hours.Methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols.Then,some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours.Real-time quantitative PCR (RT-qPCR) was performed to determine mRNA expression of Nrf2 and Bach1,Western blot analysis to measure nuclear expression of Nrf2 and Bach1 proteins,and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus.Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs,so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments.HSFs cultured without tea polyphenols served as control group.After the treatment,the 5-mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein expression of Bach 1 (mRNA:0.629 ± 0.077 vs.0.940 ± 0.033,t =6.397,P ＜ 0.05;protein:1.424 ± 0.171 vs.16.966 ± 1.702,t =15.730,P ＜ 0.05),but significantly increased mRNA and nuclear protein expression of Nrf2 (mRNA:1.467 ± 0.076 vs.0.977 ± 0.091,t =7.133,P ＜ 0.05;protein:6.929 ± 0.121 vs.3.537 ± 0.126,t =33.636,P ＜ 0.05) compared with the control group.Immunofluorescence assay showed increased accumulation of Nrf2 protein,but decreased accumulation of Bach1 protein in the nucleus.Conclusion Tea polyphenols can promote the mRNA and nuclear protein expression as well as nuclear distribution of Nrf2,but suppress the mRNA and nuclear protein expression as well as nuclear distribution of Bach 1,finally exerting antioxidative effects.