Study of the radiotherapy sensitizea-tion of retroperitoneal lymph node metastasis in patients by sodium glycididazole.

To observe the radiotherapy sensitization effect of retroperitoneal lymph node metastasis in patients by sodium glycididazole in recent clinical efficacy and toxicity. A total of 42 patients admitted with metastasis and recurrence of retroperitoneal lymph node from September 2006 to December 2009 were classified with the method of case-control. After three dimensional conformal radiation therapy with or without sodium glycididazole (800 mg/m²) for sensitization, the results of recent clinical efficacy, relief of pain, and Karnofsky score were obtained. Tumor remission rate of patients in sensitization group (with sodium glycididazole) at post-radiotherapy 3 months was significant higher than that in control group (without sodium glycididazole) (52% vs. 24%; P<0.05). Oral dose of morphine daily, and Karnofsky score in anterior-posterior radiotherapy of patients in the sensitization group were significant different with those in the control group (93 ± 12 and 42 ± 6 mg vs. 94 ± 12 and 20 ± 5 mg and (65 ± 4) and (90 ± 9) vs. (64 ± 5) and (80 ± 10), respectively; P<0.01). Sodium glycididazole has positive radiotherapy sensitization to the metastasis or recurrence of retroperitoneal lymph node for digestive tract cancer, which could obviously improve the life quality or release the pain for patients.

Correlations of IFN-γ-inducible protein-10 with the risk of chronic hepatitis B and the efficacy of interferon therapy in Asians.

PURPOSE: The aim of this study was to identify the correlations of IFN-γ-inducible protein-10 (IP-10) with the risk of chronic hepatitis B (CHB) and the efficacy of interferon therapy in Asians. METHOD: Serum IP-10 levels were assayed using enzyme linked immunosorbent assay (ELISA) in both CHB and control group. CHB group received interferon-α2b treatment to compare the pre-treatment and post-treatment serum IP-10 levels. Relevant studies met predefined inclusion and exclusion criteria were enrolled into further meta-analysis. Stata 12.0 software was applied for data analysis. RESULT: Our case-control study demonstrated that CHB group had evaluated serum IP-10 levels compared with control group (285.7 ± 41.6 pg/mL vs. 79.1 ± 33.8 pg/mL, t = 21.85, P < 0.001. After treatment for 12 weeks, CHB group had remarkably decreased post-treatment serum IP-10 levels than pre-treatment (78.5 ± 20.4 pg/mL vs. 285.7 ± 41.6 pg/mL, t = 33.76, P < 0.001). No significance was observed on post-treatment serum IP-10 levels between CHB and control group (78.5 ± 20.4 pg/mL vs. 78.1 ± 33.8 pg/mL, t = 0.07, P = 0.947). Meta-analysis results demonstrated that serum IP-10 levels in CHB group were obviously higher than healthy controls (SMD = 2.21, 95% CI = 1.55~2.87, P < 0.001). A subgroup based on the HBeAg states revealed that serum IP-10 levels in both HBeAg-positive and HBeAg-negative CHB patients were notably higher than healthy controls (HBeAg-positive: SMD = 2.00, 95% CI = 1.13-2.87, P < 0.001; HBeAg-negative: SMD = 1.34, 95% CI = 0.97-1.72, P < 0.001). CONCLUSION: Serum IP-10 may be correlated with the risk of CHB and the efficiency of interferon therapy, thus IP-10 may be a good biomarker for the diagnosis and treatment of CHB.

[Study on Speciation Analysis and Ecological Risk Assessment of Heavy Metals in Surface Sediments in Gansu, Ningxia and Inner Mongolia Sections of the Yellow River in Wet Season with HR-ICP-MS].

In order to continuously study the contents, pollution condition and potential ecological risk of heavy metals in surface sediments in Gansu, Ningxia and Inner Mongolia sections of the Yellow River in wet seasons in different years, the speciation analysis of 9 kinds of heavy metals including Cd, Pb, Cr, Ni, Cu, V, Co, Zn and Mn, pollution condition and potential ecological risk of heavy metals in surface sediments from 10 sampling sites like Baotoufuqiao (S2), Shizuishantaolezhen (S6) and Wujinxia (S9) in Gansu, Ningxia and Inner Mongolia sections of the Yellow River in 2012 wet season were studied with BCR sequential extraction and high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS) based on our previous works. The results implied that the order of heavy metals average contents in the 10 sediment samples were the same: Mn>V> Zn>Cr>Cu>Ni>Pb>Co>Cd. In the sediments, heavy metals mainly existed in the form of residual fraction, which indicated that the bioavailability or environmental impact was low. Results of geo-accumulation indices (Igeo) showed that Igeo(CD), was the largest among the heavy metals with the strongest pollution, while IGEO(Mn)was the smallest. Enrichment factor (EF) indicated that only Cd and Cu were enriched at some sampling sites. In S5, because EFcd reached 4. 69, Cd was affected by human activities obviously and the result was consistent with I. Potential ecological risk index (RI) implied that the RI values in S1, S2 and S5 were between 150 and 300, which belonged to moderate polluting degree, while others were less than 150, belonging to light pollution degree. The results of this paper could not only provide reliable experimental data and theoretical basis for the relevant departments, but also supply the technical support for constructing mathematics model of sediments-pollutants transport, systematically researching the migration and transformation rule of persistent toxic substances and environmental assessment in these reaches.

[Research on the Content Characteristics and Pollution Evaluation of Heavy Metals in Filtered Water and Suspended Particles from Gansu, Ningxia and Inner Mongolia Sections of the Yellow River in Wet Season Using HR-ICP-MS].

The content characteristics, pollution evaluation and source identification of 6 heavy Metals (Cd, Pb, Cr, As, Cu and Zn) in filtered water and 9 heavy Metals (Cd, Pb, Cr, Ni, Cu, V, Co, Zn and Mn) in suspended particles from 10 sampling sites such as Zhaojunfuqiao (S1) and Baotoufuqiao (S2), etc. from Gansu, Ningxia and Inner Mongolia sections of the Yellow River in 2012 Wet Season were studied to understand the condition of the heavy metal pollution in Gansu, Ningxia and Inner Mongolia Sections of the Yellow River by using high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS). Multivariate geochemical approaches and statistical analysis were also exploited for assessing the level of heavy metals in filtered water and suspended particles from studied area. The results showed that in filtering water, only the concentrations of Cr exceeded the standard value of Environmental Quality Standard for Surface Water (GB3838-2002) and were the highest (74.8-94.7 µg x L(-1)) among all elements in 10 sampling sites; Single factor pollution index (I(i)) results suggested that the water quality in all sampling sites were contaminated by both Cr and total nitrogen (TN), with the exception of TN in Baotoufuqiao (S2); Integrated Nemerow pollution index (I) indicated that the I values in all sampling sites were between 1-2 (light pollution), which implied that the water quality in Gansu, Ningxia and Inner Mongolia sections, especially downstream sections (S1-S6) of the Yellow River wasn't an ideal source for drinking and using in aquaculture any more. In suspended particles, concentrations of heavy metals were relatively higher than their soil background values in 10 sampling sites, except Ni in S10 (34.7 µg x L(-1)). Index of geo-accumulation (I(geo)) indicated that the I(geo) values of Pb, Cr, Ni, Cu, V, Co, Zn and Mn in all sampling sites were less than 1 (unpolluted or unpolluted-moderately polluted), respectively, while I(geo)Cd were the highest in 10 sampling sites among all heavy metals and with the moderately to strong contamination in Zhaojunfuqiao (S1), Baotoufuqiao (S2), Wuhai (S5) and Dongdagouruhuanghekou (S8). The results of this paper would help to supply reliable experimental data for researching of distribution, migration and effective protection of heavy metals in study area.

Heavy Metals Contents and Speciation in Surface Sediments from Gansu, Ningxia and Inner Mongolia Sections of the Yellow River of China.

So far, many investigations had been made on the concentration and species distribution of heavy metals in aquatic environments. However, there are only a few studies on heavy metals in upper reaches of the Yellow River, especially in Gansu, Ningxia and Inner Mongolia sections. We have literatures related to the Yellow River, in this work, we remarkably discussed about the contents, speciation and potential risks of Cd, Pb, Cr, V, Co, Ni, Cu, and Zn in surface sediments from 12 sampling sites in Gansu, Ningxia, and Inner Mongolia sections of the Yellow River of China in 2011 year wet season by high resolution inductively coupled plasma mass spectrometer (HR-ICP-MS) and sequential extraction procedure of BCR method. The results indicated that the metals contents were arranged as Cr > V > Zn > Cu > Ni > Pb > Co > Cd in all sites. Comparing with the background value of soil in local section, Cd showed the highest level at S5 (1.30 µg x g(-1)), which was almost 13 times higher than the background value (0.103 µg x g(-1)). Pollution assessment indicated that Cd presented a strong polluted status with the geo-accumulation index (I(geo)) value of 3.08 at S5, moderately to strong polluted status with the I(geo) ranged from 2.02 to 2.90 in Inner Mongolia section (S1-S4). Moreover, enrichment factor (EF) showed that all heavy metals in these sediments have been influenced by anthropogenic activities. According to potential ecological risk index (RI), S5 and S3 demonstrated high ecologic risk of heavy metals, while other sampling sites showed moderately ecological risk. The results of BCR exhibited that Cd was the most available metal, followed by Co and Ni, while V and Cr were unavailable in the sediments. Risk assessment code (RAC) exhibited high risk for Cd at S1-S4 and very high risk at S5, while medium risk for Ni and Co at all sites. The results and conclusions may be important information and therefore of interest to the relevant departments of the governments.

Regulation of T cell proliferation by JMJD6 and PDGF-BB during chronic hepatitis B infection.

T cell functional exhaustion during chronic hepatitis B virus (HBV) infection may contribute to the failed viral clearance; however, the underlying molecular mechanisms remain largely unknown. Here we demonstrate that jumonji domain-containing protein 6 (JMJD6) is a potential regulator of T cell proliferation during chronic HBV infection. The expression of JMJD6 was reduced in T lymphocytes in chronic hepatitis B (CHB) patients, and this reduction in JMJD6 expression was associated with impaired T cell proliferation. Moreover, silencing JMJD6 expression in primary human T cells impaired T cell proliferation. We found that JMJD6 promotes T cell proliferation by suppressing the mRNA expression of CDKN3. Furthermore, we have identified platelet derived growth factor-BB (PDGF-BB) as a regulator of JMJD6 expression. PDGF-BB downregulates JMJD6 expression and inhibits the proliferation of human primary T cells. Importantly, the expression levels of JMJD6 and PDGF-BB in lymphocytes from CHB patients were correlated with the degree of liver damage and the outcome of chronic HBV infection treatment. Our results demonstrate that PDGF-BB and JMJD6 regulate T cell function during chronic HBV infection and may provide insights for the treatment strategies for CHB patients.

A novel antibody humanization method based on epitopes scanning and molecular dynamics simulation.

1-17-2 is a rat anti-human DEC-205 monoclonal antibody that induces internalization and delivers antigen to dendritic cells (DCs). The potentially clinical application of this antibody is limited by its murine origin. Traditional humanization method such as complementarity determining regions (CDRs) graft often leads to a decreased or even lost affinity. Here we have developed a novel antibody humanization method based on computer modeling and bioinformatics analysis. First, we used homology modeling technology to build the precise model of Fab. A novel epitope scanning algorithm was designed to identify antigenic residues in the framework regions (FRs) that need to be mutated to human counterpart in the humanization process. Then virtual mutation and molecular dynamics (MD) simulation were used to assess the conformational impact imposed by all the mutations. By comparing the root-mean-square deviations (RMSDs) of CDRs, we found five key residues whose mutations would destroy the original conformation of CDRs. These residues need to be back-mutated to rescue the antibody binding affinity. Finally we constructed the antibodies in vitro and compared their binding affinity by flow cytometry and surface plasmon resonance (SPR) assay. The binding affinity of the refined humanized antibody was similar to that of the original rat antibody. Our results have established a novel method based on epitopes scanning and MD simulation for antibody humanization.

[Study on the contents and fractionation of rare earth elements in filtering water and suspensions in Gansu, Ningxia and Inner Mongolia Sections of Yellow River by HR-ICP-MS].

In the present paper, high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS) was applied to analyse the contents of rare earth elements (REE) in the filtering water and suspensions from 12 sampling sites in Gansu, Ningxia and Inner Mongolia Sections of Yellow River, and the fractionation of REE in suspensions was also studied. The results demonstrated that the contents of REE in filtering water were very low, and the element with highest content of elements was La(32.0 ng x L(-1)). The average concentration of total REE (sigmaREE) in filtering water was 47.5 ng x L(-1), and the Baotou-Dengkou (S1) and Sanhuhekou (S2) sampling sites had a higher sigmaREE of 130.0 and 100.0 ng x L(-1), which implied that there might be external REE sources in the filtering water. And in suspensions, the average content of sigmaREE was 167.8 mg x kg(-1), which was similar to the Chinese soil background value. While the heavy rare earth elements (HREE) content in Shenchuanqiao (S11) of Gansu province were significantly higher than other stations, which indicated that it might have external HREE sources. The chondrite-normalized patterns of REE in suspensions from the study area were similar to that of Chinese soil, with light rare earth elements enrichment, and moderate Eu depletion and a bit of Ce depletion. It was showed that the contents of REE in suspensions were mainly from soil weathering.

[Study on speciation and fractionation of rare earth eleinents in surface sediments in Gansu, Ningxia and Inner Mongolia sections of Yellow River].

In the present paper, BCR sequential extraction and high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS) were performed to analyze the speciation contents of 14 kinds of rare earth elements (REE) in the surface sediments from 12 sampling sites (S1-S12) in Gansu, Ningxia and Inner Mongolia Sections of Yellow River, and REE fractionation were also studied. The results indicated that the contents of REE in 12 sediment samples werethe same order. The average contents of 14 rare earth elements were in this order: Ce(66.4)>La(35.8)>Nd(28.6)>Pr(7.88)>Sm(5.87) >Gd(5.01)>Dy (4.53)>Yb(2.86)>Er(2.51)>Eu(1.31)>Ho(0.856)>Tb(0.760)>Tm(0.428)>Lu(0.404), which were similar to the Chinese soil background. The residual fractions of all elements were present at the highest percentages(71.9%-93.9%), which indicated that the bioavailability or environmental impact was low. The percentage of reducible fraction was the lowest, ranged from 0. 20% to 3. 87% with the mean value of 0.83%, while the oxidizable fraction percentage(7.61%) was close to acid-soluble fraction(7.69%). But in Maqu (S12), oxidizable percentage (16.1%) was significantly higher than the acid-soluble fraction (1.73%). Correlation analysis showed that there was a significant positive correlation between total organic carbon (TOC) content and oxidizable percentage, and the correlation coefficients were between 0.763 and 0.914. REE fractionation results showed that: the contents of REE in surface sediments of Gansu, Ningxia and Inner Mongolia Sections of Yellow River were mainly from soil weathering, with light-REE enrichment and Eu depletion. The chondrite-normalized curve implied that La and Ce in Jinshawan (S8) and Baotou-Dengkou (S1) and heavy REE at all the sampling points might have external REE sources.

A clinical trial of acupuncture about time-varying treatment and points selection in primary dysmenorrhea.

OBJECTIVE: To observe the clinical therapeutic effects of acupuncture at single point Shiqizhui (EX-B8) and multi-points in time-varying treatment for primary dysmenorrhea. METHODS: 600 patients with primary dysmenorrhea were randomly assigned to the single point group (n=200) including group A (treating before the menstruation, n=100) and group B (immediately treating as soon as pain occurrence, n=100), the multi-points group (n=200) including group C (treating before the menstruation, n=100) and group D (immediately treating as soon as pain occurrence, n=100), or the control group, group E (n=200, no treatment). The therapeutic effects were analyzed after treatment for three menstrual cycles and interviewed for three follow-up periods. RESULTS: Acupuncture could effectively relieve menstrual pain for primary dysmenorrhea compared with the control group (P<0.05, P<0.01). Immediate pain relief occurred following acupuncture within 5 min in group B (P<0.01) and group D (P<0.01), and the two groups obviously relieved menstrual pain for VAS scores. Both group A and group C obviously relieved menstrual pain (P<0.01), and group C was better than group A (P<0.05). Compared with group D, Group C was much better for CMSS scores in cycle 1. CONCLUSION: Treating before the menstruation is better than immediately treating as soon as pain occurrence at the improvement in symptoms of dysmenorrheal at multi-points. And single point is better than multi-points when immediately treating as soon as pain occurrence. The present trial suggest Shiqizhui (EX-B8) should be chosen as a convenient point.

Does Traditional Chinese Medicine pattern affect acupoint specific effect? Analysis of data from a multicenter, randomized, controlled trial for primary dysmenorrhea.

OBJECTIVES: This study assessed the importance of the Traditional Chinese Medicine (TCM) pattern on an acupoint-specific effect. DESIGN: This was a TCM pattern subdivision analysis of the first intervention data from a multicenter, randomized, controlled trial (ISRCTN24863192) (the main trial). SETTINGS: The main trial recruited participants from six hospitals in three provinces in China. SUBJECTS: Five hundred and one (501) participants diagnosed with primary dysmenorrhea (PD) were enrolled in the main trial. INTERVENTIONS: The main trial randomly and equally divided participants into three treatment groups with bilateral electroacupuncture at three sites, respectively: Sanyinjiao (SP6), Xuanzhong (GB39), and an adjacent nonacupoint. Participants were diagnosed with TCM patterns before the treatment. The intervention was carried out when the visual analogue scale (VAS) score of participant's menstrual pain was ≥ 40 mm on the first day of menstruation and lasted for 30 minutes. OUTCOME MEASURES: The immediate improvement of pain was measured with a 100-mm VAS before the intervention, at 5 minutes, 10 minutes, and 30 minutes during the intervention, and at 30 minutes after the completion of this intervention. RESULTS: Three (3) TCM patterns (n=320) were eligible for analysis, including Cold and Dampness Stagnation pattern (n=184), Qi and Blood Stagnation pattern (n=84), and Qi and Blood Deficiency pattern (n=52). In Cold and Dampness Stagnation pattern, the SP6 group had a significant reduction in VAS scores compared with the GB39 group (mean difference -7.6 mm) and the nonacupoint group (mean difference -8.2 mm), respectively. There was no difference between the latter two groups. There were no group differences in VAS scores in the other two patterns. CONCLUSIONS: It suggested that TCM pattern might affect acupoint specific effect on the immediate pain relief obtained for participants with PD.

Study on heavy metals and ecological risk assessment from Gansu, Ningxia and Inner Mongolia sections of the Yellow River, China.

The Yellow River is the most important resource of water supply in northern China. The purpose of this work are to investigate the concentrations and potential ecological risk of heavy metals in the upper reaches of the Yellow River, the concentrations of eight heavy metals including As, Hg, Cd, Pb, Cr, Ni, Cu and Zn in filtered water and suspended particles from 12 sampling sites of Gansu, Ningxia and Inner Mongolia sections of the Yellow River of China were studied by atomic fluorescence spectrometry (AFS) and high resolution inductively coupled plasma mass spectrometer (HR-ICP-MS) in this paper. The results implied that all heavy metals in filtered water were lower than the limit standards for drinking water except for Cr (56.9 approximately 71. 5 microg L-1 ). Water quality parameters such as total nitrogen (TN), total phosphorus (TP) and pH were also determined and the contents were low along the river except for TN at S1 (2.48) and S9 (2.38), which exceeded the maximum permitted concentration of Class V for the protection of surface water. In suspended particles, the concentrations of Hg, Cd, Pb and Zn were much higher than those in the background value of soil from local section. Cluster analysis (CA) indicated that same sources for Ni, Cu, Cr, Zn and Pb could be stainless steel and petrochemical industrial activities, while As, Cd and Hg derived from agrochemicals, fertilizers, mining, fuel and coal combustion, respectively. Ecological risk assessment was undertaken using risk index (RI) for sampling sites and ecological risk factor (Er) for heavy metals. Eleven suspension samples existed considerable ecological risk (300.6< RI< 508. 6), while S1 was moderate ecological risk (RI, 299.3). According to Er, Hg had considerable or high ecological risk in Inner Mongolia section, while very high ecological risk for Cd at S11 (396.0), S9 (384. 0) and S5 (373. 3), respectively, implied a high pollution in these sampling sites. The results could provide reliable experimental data and theoretical basis for the relevant departments.

Structure-based high-throughput epitope analysis of hexon proteins in B and C species human adenoviruses (HAdVs).

Human adenoviruses (HAdVs) are the etiologic agent of many human infectious diseases. The existence of at least 54 different serotypes of HAdVs has resulted in difficulties in clinical diagnosis. Acute respiratory tract disease (ARD) caused by some serotypes from B and C species is particularly serious. Hexon, the main coat protein of HAdV, contains the major serotype-specific B cell epitopes; however, few studies have addressed epitope mapping in most HAdV serotypes. In this study, we utilized a novel and rapid method for the modeling of homologous proteins based on the phylogenetic tree of protein families and built three-dimensional (3D) models of hexon proteins in B and C species HAdVs. Based on refined hexon structures, we used reverse evolutionary trace (RET) bioinformatics analysis combined with a specially designed hexon epitope screening algorithm to achieve high-throughput epitope mapping of all 13 hexon proteins in B and C species HAdVs. This study has demonstrated that all of the epitopes from the 13 hexon proteins are located in the proteins' tower regions; however, the exact number, location, and size of the epitopes differ among the HAdV serotypes.

Downregulation of the AU-rich RNA-binding protein ZFP36 in chronic HBV patients: implications for anti-inflammatory therapy.

Inflammation caused by chronic hepatitis B virus (HBV) infection is associated with the development of cirrhosis and hepatocellular carcinoma; however, the mechanisms by which HBV infection induces inflammation and inflammatory cytokine production remain largely unknown. We analyzed the gene expression patterns of lymphocytes from chronic HBV-infected patients and found that the expression of ZFP36, an AU-rich element (ARE)-binding protein, was dramatically reduced in CD4(+) and CD8(+) T lymphocytes from chronic HBV patients. ZFP36 expression was also reduced in CD14(+) monocytes and in total PBMCs from chronic HBV patients. To investigate the functional consequences of reduced ZFP36 expression, we knocked down ZFP36 in PBMCs from healthy donors using siRNA. siRNA-mediated silencing of ZFP36 resulted in dramatically increased expression of multiple inflammatory cytokines, most of which were also increased in the plasma of chronic HBV patients. Furthermore, we found that IL-8 and RANTES induced ZFP36 downregulation, and this effect was mediated through protein kinase C. Importantly, we found that HBsAg stimulated PBMCs to express IL-8 and RANTES, resulting in decreased ZFP36 expression. Our results suggest that an inflammatory feedback loop involving HBsAg, ZFP36, and inflammatory cytokines may play a critical role in the pathogenesis of chronic HBV and further indicate that ZFP36 may be an important target for anti-inflammatory therapy during chronic HBV infection.

Enhancement of humoral and cellular immunity in mice against Japanese encephalitis virus using a DNA prime-protein boost vaccine strategy.

A synthetic multi-epitope gene containing critical epitopes of the Japanese encephalitis virus (JEV) envelope gene was cloned into both prokaryotic and eukaryotic expression vectors. The recombinant plasmid and purified recombinant protein (heterologously expressed in Escherichia coli) were used as immunogens in a mouse model. The results indicate that both the recombinant protein and the DNA vaccine induce humoral and cellular immune responses. Neutralising antibody titres in mice in the pcDNA-TEP plus rEP group increased considerably relative to mice immunised using either pcDNA-TEP or rEP alone (P<0.05). Furthermore, the highest levels of interleukin (IL)-2, interferon-gamma and IL-4 were induced following priming with the DNA vaccine and boosting with the recombinant protein. Together these findings demonstrate that a DNA-recombinant protein prime-boost vaccination strategy can produce high levels of antibody and trigger significant T cell responses in mice, highlighting the potential value of such an approach in the prevention of JEV infection.

Immune responses of recombinant adenoviruses expressing immunodominant epitopes against Japanese encephalitis virus.

Japanese encephalitis virus (JEV), which belongs to the family Flaviviridae, causes infection of the central nervous system in humans and equines and stillbirths in swine. In the present report, we constructed and characterized the immune responses conferred by recombinant adenoviruses expressing JEV E epitopes (six amino acid residues 60-68, 327-333, 337-345, 373-399, 397-403 and 436-445 in E, designated TEP). Seven groups (n = 10) of female BALB/c mice received intramuscular (IM) or oral immunization with the recombinant adenoviruses twice at 2-week intervals. Intramuscular immunization of mice with rAd-TEP generated greater titers of anti-JEV antibodies and JEV neutralizing activity than in animals with oral injection. It statistically significant differences were found in anti-JEV antibody titers and JEV neutralizing activity induced by IM immunization with rAd-TEP at a dose of 1 x 10(8.0)TCID50 when compared with the doses tested (3 x 10(7.0) and 1 x 10(7.0)TCID50) IM inoculation of rAd-TEP. Splenocytes from mice immunized intramuscularly with rAd-TEP secreted the largest amounts of interferon-gamma and interleukin-2 and moderate amounts of interleukin-4 in the presence of JEV. It demonstrates that IM immunization with rAd-TEP induced the highest level of cell-mediated immune responses and the higher level of JEV-specific humoral immune responses than oral immunization. Then we further evaluated the protective efficacy of the recombinants in swine. All swine were protected from viral challenge with IM rAd-TEP at 1 x 10(10.0)TCID50, even though the neutralizing antibody titers were lower than those in the group inoculated with inactivated vaccine. Our findings indicate that rAd-TEP might be an attractive candidate vaccines for preventing JEV infection.