Deletion of Luzp2 Does Not Cause Hearing Loss in Mice.

Deafness is the prevailing sensory impairment among humans, impacting every aspect of one's existence. Half of congenital deafness cases are attributed to genetic factors. Studies have shown that Luzp2 is expressed in hair cells (HCs) and supporting cells of the inner ear, but its specific role in hearing remains unclear. To determine the importance of Luzp2 in auditory function, we generated mice deficient in Luzp2. Our results revealed that Luzp2 has predominant expression within the HCs and pillar cells. However, the loss of Luzp2 did not result in any changes in auditory threshold. HCs or synapse number and HC stereocilia morphology in Luzp2 knockout mice did not show any notable distinctions. This was the first study of the role of Luzp2 in hearing in mice, and our results provide important guidance for the screening of deafness genes.

Differences in gut microbiota and its metabolic function among different fasting plasma glucose groups in Mongolian population of China.

BACKGROUND: Many studies reported the association between gut microbiota and type 2 diabetes mellitus (T2D), but it is still unclear which bacterial genus plays a key role and how the metabolic function of gut microbiota changes in the occurrence and development of T2D. Besides, there is a high diabetic prevalence in Mongolian population, which may be partly affected by their high calorie diet. This study identified the main bacterial genus influencing T2D in Mongolian population, and analyzed the changes of metabolic function of gut microbiome. The association between dietary factors and the relative abundance of main bacterial genus and its metabolic function was also studied. METHODS: Dietary surveys and gut microbiota test were performed on 24 Mongolian volunteers that were divided into T2D (6 cases), PRET2D (6 cases) and Control group (12 cases) according to fasting plasma glucose (FPG) values. The relative abundance and metabolic function of gut microbiome from their fecal samples were measured by metagenomic analysis. Statistic method was used to evaluate the association between dietary factors and the relative abundance of the main bacterial genus or its metabolic function. RESULTS: This study found that the Clostridium genus may be one of the key bacterial genera affecting the process of T2D. First, the relative abundance of Clostridium genus was significantly different among the three groups. Second, there was a higher relative abundance of metabolic enzymes of gut bacteria in PRET2D and T2D group than that in Control group. Third, a strong correlation between Clostridium genus and many metabolic enzymes was uncovered, many of which may be produced by the Clostridium. Last, carotene intake daily was negatively correlated with the Clostridium but positively correlated with tagaturonate reductase catalyzing interconversions of pentose and glucuronate. CONCLUSIONS: The gut Clostridium genus may play an important role in the development of T2D and it could be a potential biomarker for T2D in Mongolian population. Meanwhile, the metabolic function of gut bacteria has changed during the early stage of T2D and the changes in carbohydrate, amino acid, lipid or energy metabolism of Clostridium genus may play a critical role. In addition, the carotene intake may affect reproduction and metabolic function of Clostridium genus.

Hypoxia-induced Circular RNA hsa_circ_0006508 Promotes the Warburg Effect in Colorectal Cancer Cells

Background: The hypoxia-induced Warburg effect promotes colorectal cancer malignancy with altered circular RNA (circRNA) expression. Aims: To investigate the association with the Warburg effect in colorectal cancer and whether has_circ_0006508 can be induced by hypoxia. Study design: In vitro cell lines and human-sample study. Methods: The biological functions of circ_0006508 and miR-1272 in the viability, colony formation, and glycolysis under hypoxic conditions were determined by loss-of-function and gain-of-function experiments. The chromatin immunoprecipitation assay was used to demonstrate the direct binding between circ_0006508 promoters and hypoxia-inducible factor 1α (HIF-1α). Transcription activity was subjected to the Luciferase reporter assay. The correlation of circ_0006508 and miR-1272 with overall survival was determined with the Kaplan-Meier analysis. Results: Upregulated circ_0006508 and downregulated miR-1272 were observed in colorectal cancer samples, which was associated with the TNM stage and overall survival. Functional assays demonstrated that the hypoxia-induced upregulated circ_0006508 and downregulated miR-1272 promoted the viability and Warburg effect of colorectal cancer in vitro. Mechanistically, HIF-1α-induced circ_0006508 could directly sponge miR-1272, which played a suppressive role in glycolysis. Conclusion: Circ_0006508-mediated miR-1272 inhibition could promote the malignant behaviors of colorectal cancer with an upregulated Warburg effect.

Determining the safety and effectiveness of Tai Chi: a critical overview of 210 systematic reviews of controlled clinical trials.

BACKGROUND: This overview summarizes the best available systematic review (SR) evidence on the health effects of Tai Chi. METHODS: Nine databases (PubMed, Cochrane Library, EMBASE, Medline, Web of Science, China National Knowledge Infrastructure (CNKI), Chinese Scientific Journal Database (VIP), Sino-Med, and Wanfang Database) were searched for SRs of controlled clinical trials of Tai Chi interventions published between Jan 2010 and Dec 2020 in any language. Effect estimates were extracted from the most recent, comprehensive, highest-quality SR for each population, condition, and outcome. SR quality was appraised with AMSTAR 2 and overall certainty of effect estimates with the GRADE method. RESULTS: Of the 210 included SRs, 193 only included randomized controlled trials, one only included non-randomized studies of interventions, and 16 included both. Common conditions were neurological (18.6%), falls/balance (14.7%), cardiovascular (14.7%), musculoskeletal (11.0%), cancer (7.1%), and diabetes mellitus (6.7%). Except for stroke, no evidence for disease prevention was found; however, multiple proxy-outcomes/risks factors were evaluated. One hundred and fourteen effect estimates were extracted from 37 SRs (2 high, 6 moderate, 18 low, and 11 critically low quality), representing 59,306 adults. Compared to active and/or inactive controls, 66 of the 114 effect estimates reported clinically important benefits from Tai Chi, 53 reported an equivalent or marginal benefit, and 6 an equivalent risk of adverse events. Eight of the 114 effect estimates (7.0%) were rated as high, 43 (37.7%) moderate, 36 (31.6%) low, and 27 (23.7%) very low certainty evidence due to concerns with risk of bias (92/114, 80.7%), imprecision (43/114, 37.7%), inconsistency (37/114, 32.5%), and publication bias (3/114, 2.6%). SR quality was often limited by the search strategies, language bias, inadequate consideration of clinical, methodological, and statistical heterogeneity, poor reporting standards, and/or no registered SR protocol. CONCLUSIONS: The findings suggest Tai Chi has multidimensional effects, including physical, psychological and quality of life benefits for a wide range of conditions, as well as multimorbidity. Clinically important benefits were most consistently reported for Parkinson's disease, falls risk, knee osteoarthritis, low back pain, cerebrovascular, and cardiovascular diseases including hypertension. For most conditions, higher-quality SRs with rigorous primary studies are required. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42021225708.

Analysis of Dietary Supplement Use and Influencing Factors in the Mongolian Population.

Objective: Dietary supplements (DS) may improve micronutrient deficiencies, but the unique eating habits and cultural customs of the Chinese Mongolian population affect their choice of DS. Therefore, this study adopted a cross-sectional method to explore the current status of DS use and to assess the influencing factors in the Mongolian population in Inner Mongolia, China. Methods: We used a multistage random cluster sampling method to select 1,434 Mongolian people aged ≥ 18 years in Hohhot and Xilinhot, Inner Mongolia. Data regarding general patient characteristics and DS use through questionnaire surveys were obtained, and the blood plasma was collected for biochemical index detection. The binary logistic regression and decision tree algorithm were used to predict the factors influencing DS use among the Mongolian population. Results: Among 1,434 participants that completed the baseline survey, the usage rate of DS was 18.83%, and more women than men used DS (P = 0.017). Higher use of DS was reported among individuals aged ≤ 34 years, but this difference is not statistically significant (P = 0.052). Usage rate was higher among those living in urban areas (P < 0.001), those with higher education (P < 0.001), those engaged in mental work (P < 0.001), and nonsmokers (P = 0.019). The biochemical test results showed that the proportion of people with abnormal total cholesterol levels using DS was lower (P = 0.003), but that of those with abnormal triglyceride levels using DS was higher (P = 0.001), compared with the proportion of those with normal levels in each case. The most commonly used supplement was calcium (58.15%). Education level was the main factor affecting DS intake. The results of the binary logistic regression model and decision tree model both showed that region, educational level, and abnormal triglyceride levels were significant factors influencing DS intake among Mongolians. Conclusion: Findings from this study indicate that DS intake is uncommon in the Mongolian population. In addition, sex, region, education level, and triglyceride levels may influence DS use.

Tai Chi for health and well-being: A bibliometric analysis of published clinical studies between 2010 and 2020.

The objective of this bibliometric review was to identify the volume, breadth, and characteristics of clinical studies evaluating Tai Chi published between January 2010 and January 2020. Five English and four Chinese language databases were searched. Following independent screening, 1018 eligible publications representing 987 studies were identified, which was a three-fold increase from the previous decade. Most common were randomized controlled trials (548/987, 55.5 %), followed by systematic reviews (157/987, 15.9 %), non-randomized controlled clinical studies (152/987, 15.4 %), case series (127/987, 12.9 %) and case reports (3/987, 0.3 %) that were conducted in China (730/987, 74.0 %), followed by the United States of America (123/987, 12.5 %) and South Korea (20/987, 2.0 %). Study participants were mostly in the adult (55.2 %) and/or older adult (72.0 %) age groups. The top ten diseases/conditions were hypertension, chronic obstructive pulmonary disease, diabetes, knee osteoarthritis, heart failure, depression, osteoporosis/osteopenia, breast cancer, coronary heart disease and insomnia. A quarter of the studies enrolled healthy participants to evaluate the effects of Tai Chi on health promotion/preservation, balance/falls, and physiological/biomechanical outcomes. Yang style Tai Chi was the most popular, followed by Chen and Sun style. Tai Chi was mostly commonly delivered face-to-face by a Tai Chi instructor in group settings for 60 min, three times a week, for 12 weeks. Most studies (93.8 %) reported at least one outcome in favor of Tai Chi. Adverse events were underreported (7.2 %). Over half fell short of expected intervention reporting standards, signalling the need for Tai Chi extensions to existing guidelines.

Eating behaviors related to brucellosis infection in Mongolian middle school students in Abaga Banner, Inner Mongolia / 中国学校卫生

Objective@#To understand current situation and associated factors of brucellosis infection among students in Mongolian middle school in Abaga Banner, and to provide basis for further development of prevention and treatment strategies.@*Methods@#Serological examinations were performed on pupils and middle school students in a Mongolian middle school in Abaga Banner. Questionnaire surveys were used to investigate students knowledge of eating behaviors related to brucellosis infection.@*Results@#A total of 500 primary and middle school students were investigated, and 6 cases of brucellosis were confirmed, and the infection rate was 1.20%. There were 4 cases of male infection and 2 cases of female infection. The infection rate in boys (1.61%) was higher than that of girls (0.79%). Higher proportion was found in 14-year-old group (n=3), accounting for 17.40%. The residence period of 10-15 years includes 6 cases of all infected persons. The number of infections in Narenbaolige Town was 4, accounting for 23.40%. Univariate logistic regression analysis shows that frequent consumption of unsterilized dairy products, milking, processing lambs or slugs and infected cloth The disease was positively correlated (OR=11.80, 139.14, 8.02, P<0.05). Multivariate Logistic regression analysis showed that milking was positively correlated with brucellosis infection(OR=20.19, 95%CI=2.01-202.74, P=0.01).@*Conclusion@#The brucellosis infection of Mongolian primary and middle school students in Abaga Banner is related to a variety of related dietary factors, and its prevention and treatment should be strengthened.

Activating molecular oxygen with Au/CeO2 for the conversion of lignin model compounds and organosolv lignin.

Au/CeO2 was demonstrated to be a high efficiency catalyst for the conversion of 2-phenoxyacetophenol (PP-ol) employing O2 as an oxidant and methyl alcohol as the solvent without using an erosive strong base or acid. Mechanistic investigations, including emission quenching experiments, electron spin-resonance (ESR) and intermediate verification experiments, were carried out. The results verified that the superoxide anion activated by Au/CeO2 from molecular oxygen plays a vital role in the oxidation of lignin model compounds, and the cleavage of both the ß-O-4 and Cα-Cß linkages was involved. Au/CeO2 also performed well in the oxidative conversion of organosolv lignin under mild conditions (453 K), producing vanillin (10.5 wt%), methyl vanillate (6.8 wt%), methylene syringate (3.4 wt%) and a ring-opened product. Based on the detailed characterization data and mechanistic results, Au/CeO2 was confirmed to be a promising catalytic system.

Mir-22-3p Enhances the Chemosensitivity of Gastrointestinal Stromal Tumor Cell Lines to Cisplatin through PTEN/PI3K/Akt Pathway.

Mir-22-3p is associated with many important biological processes, including neuroprotection, tumorigenesis, and various other tumor progressions. Our study aimed to investigate the roles of Mir-22-3p in chemosensitivity of gastrointestinal stromal tumor (GIST-T1) cells to cisplatin and explore its underlying mechanisms. Mir-22-3p high-expressing cell line was established by transfecting GIST-T1 cell line cells with Mir-22-3p mimic. After treatment with cisplatin (10 µM), Cell counting kits-8 (CCK-8) method was used to detect the cell viability. Flow cytometry was applied to measure the degree of cell apoptosis. Scratch wound healing test was used to detect the migration ability of cells. The protein and mRNA levels of the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) pathway-related factors were analyzed by Western blot and qRT-PCR. The mRNA level of Mir-22-3p was increased in transfected GIST-T1 cells compared with that in control cells. The survival rate and Bcl-2/Bax ratio of GIST-T1 cells treated with both Mir-22-3p analogue and cisplatin were significantly decreased, while the apoptosis rate and protein level of caspase-3 were significantly increased (p<0.05). In addition, the mRNA and protein levels of PTENwere significantly increased in cells treated with both Mir-22-3p analogue and cisplatin (p<0.05), while the expression levels of PI3K and Akt were significantly decreased (p<0.05). Mir-22-3p overexpression can increase the chemosensitivity of cisplatin in human gastrointestinal stromal tumor cells by PTEN/PI3K/Akt pathway.

Determination of phthalate esters in airborne particulates by heterogeneous photo-Fenton catalyzed aromatic hydroxylation fluorimetry.

The environmental contaminants phthalic acid esters (PAEs) were determined by aromatic hydroxylation fluorimetry combined with heterogeneous photo-Fenton process in the presence of vermiculite supported BiFeO3 (VMT-BiFeO3). In strong alkaline solution, PAEs were hydrolyzed into phthalates with no fluorescence, which then reacted with hydroxyl free radicals produced in photo-Fenton process catalyzed by VMT-BiFeO3 to form the fluorescent hydroxyl phthalates. The fluorescence intensity was proportional to the concentration of PAEs with the maximum excitation and emission wavelength of 300nm and 417nm, respectively. A good linear relationship can be obtained in the range of 3.8×10-7 to 4.8×10-5molL-1 for DEP with correlation coefficient of 0.9997, and the sensitivity of the method was high with detection limit of 5.43×10-8molL-1. The method has been successfully applied to determine total PAEs in airborne particulates with satisfactory results.

Causes of Death in Children Aged < 15 Years in the Inner Mongolia Region of China, 2008-2012.

The objective of our study was to identify the causes of death in children <15 years of age in Inner Mongolia and to examine the age-specific causes of death. Study data from 2008-2012 were obtained from the Death Registry System that is maintained by the Inner Mongolia Centers for Disease Control and Prevention. The mortality rate (per 100,000) for children <15 years of age was calculated and stratified by age in different years. We computed the proportion of age-specific causes of death for children <15 years that occurred between 2008 and 2012 across eight monitoring points in Inner Mongolia. We used a log-linear model to analyze the year and age effects on childhood mortality. From 2008-2012, the standardized mortality of children <15 years of age was 42.78/100,000. The mortality rate was not significantly different from 2008 to 2012 (p>0.05); the mortality rate was the highest in the <1-year age group (p<0.05); and the mortality rate of the <1-year age group was higher in 2012 compared to that in 2009 (p<0.05), 2010 (p<0.05), and 2011 (p<0.05). In children aged 1-14 years, the leading cause of death was injuries, among which transport accident injuries were the most prevalent. To reduce the childhood mortality rate in Inner Mongolia, China, we should focus on the prevention of perinatal deaths in infants <1 year of age and on the prevention of transport accident injuries among older children (1-14 years).

Gender differences in irritable bowel syndrome among medical students at Inner Mongolia Medical University, China: a cross-sectional study.

There is little epidemiological research on Irritable bowel syndrome (IBS) in Inner Mongolia, China. Here we investigated the prevalence of IBS and factors associated with IBS in both males and females in Inner Mongolia Medical University by a cross-sectional study. We recruited Inner Mongolia Medical University students residing in campus and asked them to complete a self-administered questionnaire. The prevalence of IBS in each factor we chose in all, male, and female students was determined. We assessed IBS subtypes in male and female students. Multivariate logistic regression models were used to assess the factors associated with IBS in male and female students. The overall prevalence of IBS was 29.5%. The prevalence of IBS in female students was significantly higher than that in male students (31.3% vs. 24.8%, p < .001). Logistic regression results showed that attempting to lose weight and anxiety were both associated with increasing odds of IBS, while exercise was not associated with IBS in either male or female students. In female students, snack consumption and depression were also both associated with increasing odds of IBS. The predominant IBS subtype was the diarrhea-predominant type in both male and female students. Considering the high prevalence of IBS in students and the fact that the factors associated with IBS can be improved by individuals, students should be given adequate education and counseling to improve their mental health and lifestyle, especially female students in higher grades.

Tubeimoside-1 (TBMS1) inhibits lung cancer cell growth and induces cells apoptosis through activation of MAPK-JNK pathway.

Tubeimoside-1 (TBMS1) is a natural compound isolated from tubeimoside, which has anti-tumor properties in some cancer cells, but its mechanisms are unclear. In the present study, we determined if TBMS1 would inhibit cell growth of human lung cancer cell lines. We found that TBMS1 inhibited growth in A549 and PC9 human lung cancer cell. Flow cytometry revealed TBMS1 arrested the cells in the G2/M phase and induced cell apoptosis. Furthermore, results from Western blotting and real-time PCR indicated that decreased the cell proliferation and cell growth-associated protein levels, such as p21, p15 and cyclin B1, TBMS1 up-regulated proapoptotic bax and cleavage of procaspase-3, down-regulated antiapoptotic Mcl-1 and cIAP-1, but did not change expression of PARP and procaspase-8. And TBMS1 also found to increase the phosphorylation, of JNK and p38, suggesting TBMS1 could activate the MAPK-JNK signaling pathway. Luciferase reporter assay revealed that TBMS1 reduced the promoter activity of AP-1, NF-κB and TNFα. In addition, TBMS1 caused the production of reactive oxygen species (ROS). These results provide evidence that TBMS1 may have potential as a novel anti-cancer agent for the treatment of lung cancer. TBMS1 inhibited cell proliferation may through MAPK-JNK signaling pathway.

Improvement effect of Lycium barbarum polysaccharide on sub-health mice.

OBJECTIVES: Sub-health has been described as a chronic condition of unexplained deteriorated physiological function, which falls between health and illness. In the present study, we evaluated the effects of Lycium barbarum polysaccharide (LBP), a polysaccharide fraction purified from Lycium barbarum (L. barbarum) on the sub-health mice. MATERIALS AND METHODS: The sub-health model mice were built through compound factors. The mice were given intragastric administration of LBP at low dose (50 mg•kg-1) and high dose (100 mg•kg-1), respectively. After LBP treatment for 4 weeks, the antioxidant ability, enhancing immune function and anti-fatigue activity were detected. RESULTS: The results showed that LBP could enhance antioxidant ability in sub-health mice. LBP could effectively improve immunity of sub-health mice and protect the immune organs, such as thymus. In addition, LBP showed anti-fatigue ability in sub-health mice. CONCLUSION: LBP could improve sub-health state caused from composite factor through three aspects, such as increasing antioxidant ability, promoting T lymphocyte proliferation, inhibiting thymus lymphocyte apoptosis, and alleviating fatigue.

Co-delivery of LIGHT expression plasmid enhances humoral and cellular immune responses to HIV-1 Nef in mice.

The immunogenicity and efficacy of a DNA vaccine can be greatly enhanced when a gene adjuvant is used. LIGHT, a member of TNF superfamily, can function as a costimulatory molecule for human naïve T cells to proliferate and can be a potential gene adjuvant. In the current study, the eukaryotic expression plasmid pcDNA-nef was constructed by inserting a full-length nef gene into pcDNA3.1(+), and an in vitro transfection experiment suggested that the nef gene could be expressed successfully in mammalian cells. BALB/c mice were immunized with HIV-1 nef DNA vaccine plasmids alone or in combination with LIGHT expression plasmids, and the specific humoral and cellular immune responses were measured. The data showed that HIV-1 nef DNA vaccine plasmids could induce anti-Nef antibodies, Nef-specific lymphocyte proliferation and CTL activity, whereas stronger specific immune responses were induced in mice when co-immunizing with HIV-1 nef DNA vaccine plasmids and LIGHT expression plasmids, suggesting that the eukaryotic expression vector encoding HIV-1 nef is capable of inducing specific immune responses towards HIV-1 Nef and that LIGHT could be considered as a gene adjuvant for HIV-1 DNA vaccination.

Gender comparisons of physical fitness indexes in Inner Mongolia medical students in China.

INTRODUCTION: The aim of present study was to investigate gender differences in physical fitness indexes in regard to BMI (body mass index) levels among Inner Mongolia medical students in China. METHODS: Data on participant characteristics came from basic information contained in the school database. Physical fitness indexes including BMI, vital capacity index, sidestep test, and standing long jump, were conducted. RESULTS: Female students had a higher rate of normal weight than those of males. The obesity rate of males was 5 times higher compared to females. Compared with male students, female students had a higher pass rate in vital capacity index, sidestep and standing long jump. Females were higher 17% than males in the pass rate of the sidestep test. Males performed better than females in the standing long jump. In both the malnutrition and normal weight group, the pass rate of the 3 physical fitness indexes for both male and female students was higher than obese group. The not pass rate was higher than pass rate both male and female students in the vital capacity index in the obese group. DISCUSSION: Males had a poor physical fitness level compared with females. Male students may be more likely to spend more time using computers and it will cut down the time of participating in physical activities. So, in our university, more attention should pay on physical education, especially for males.

Improvement in efficacy of DNA vaccine encoding HIV-1 Vif by LIGHT gene adjuvant.

DNA vaccine can induce the prolonged immune responses against the encoded antigen with the appropriate adjuvant. To study the immunogenicity of the HIV-1 vif DNA vaccine in inducing the humoral and cellular immune responses and the immunoadjuvant effect of LIGHT, which is a member of TNF superfamily and can stimulate the proliferation of naïve T cells as a co-stimulatory molecule, DNA vaccine plasmid pcDNA-Vif was constructed by inserting HIV-1 vif gene into the downstream of CMV promoter in eukaryotic expression vector pcDNA3.1(+). In vitro expression of HIV-1 Vif in pcDNA-Vif-transfected HeLa cells was confirmed in transcriptional and protein level by RT-PCR and Western blot, respectively. After BALB/c mice were injected muscularly with DNA vaccines for three times, the specific immune responses were analyzed. The data showed that anti-Vif antibody response, Vif-specific T cell proliferation, and CTL activities were induced in the mice that were inoculated with HIV-1 vif DNA vaccine plasmid. Interestingly, stronger humoral and cellular immune responses were detected in mice that were immunized with plasmid pcDNA-Vif and pcDNA-LIGHT together compared to the single immunization with plasmid pcDNA-Vif alone. Together, the results of the study suggest that candidate HIV-1 DNA vaccine can elicit HIV-1 Vif-specific immune responses in mice and that LIGHT plays the role of immunoadjuvant in co-immunization with DNA vaccine.

The role of small molecular weight compounds to increase vacuolation induced by VacA toxin in vitro.

VacA is a vacuolation protein toxin secreted by Helicobacter pylori. Many compounds have been implicated in the regulation of VacA toxin activity. In this study, regulation of cell vacuolation induced by VacA was observed with the addition of glycine, glycine hydrochloride, xylitol, and taurine by neutral red dye uptake assay using gastric human epithelial cell cultures. Glycine, xylitol, and taurine increased cell vacuolation significantly after 48h (p<0.05), with their effect apparent in a wide concentration range (0.2mM to about 100mM). Changes were sharp in respect of concentration and showed little dose-response characteristics. In contrast, upregulation of glycine hydrochloride on cell vacuolation in weak acidic extracellular pH was much retarded with VacA activity not initiated until 72h. In addition, our results showed that cell vacuolation was highest when the pH was 6.8. The increase in vacuolation was gentle in weak acidic extracellular pH and the increase dose-dependent with a Pearson correlation coefficient (r) of 0.986 from 0.2 to 6.25mM. In this concentration range and at the same time point, the pH decrease was negatively correlated with vacuolating activity (r=0.922, p<0.01). In conclusion, our study showed that three small molecular compounds can increase vacuolation induced by VacA toxin in vitro.

[Cloning and expression of the extracellular region of human LIGHT gene in E.coli.].

AIM: To construct a recombinant prokaryotic expression vector containing the extracellular region of human LIGHT gene and perform the express in E.coli. METHODS: Total RNA was extracted from human immature bone marrow-derived dendritic cells. The extracellular region of human LIGHT gene was amplified by RT-PCR and cloned into pET32a(+) vector, the recombinant plasmid was identified by restriction endonuclease digestion analysis and DNA sequencing. After the recombinant plasmid was transformed into E.coli BL21 and induced with IPTG, the expressed protein was analyzed by SDS-PAGE and Western blot. RESULTS: A 543 bp of the extracellular region of human LIGHT gene was obtained and the sequence was confirmed correct by DNA sequencing. SDS-PAGE and Western blot analysis showed that a protein with molecular weight of 41 000 was expressed in E.coli BL21. CONCLUSION: The extracellular region of LIGHT gene is cloned successfully and expressed in E.coli BL21 and the elementary expression conditions were obtained, which lays a basis on the further functional research of LIGHT.

B7-DC-silenced dendritic cells induce stronger anti-HBV immunity in transgenic mice.

Hepatitis B virus (HBV) is a noncytopathic DNA virus and is the pathogen of acute and chronic hepatitis. Interferon and nucleotide analogues such as lamivudine and adefovir are the current treatment strategies of HBV infection; however, it is still a serious disease. Therefore, the development of new therapeutic options against HBV is needed. In the present study, we have investigated whether the vectors carrying short hairpin RNA (shRNA) targeting the murine B7-DC gene could silence the expression of B7-DC and analyzed the function of gene-modified dendritic cells (DCs) by mixed lymphocyte reaction. The results demonstrated that two shRNA vectors efficiently suppressed the expression of B7-DC. The MLR assay showed that shRNA-B7-DC-transfected DCs induced markedly higher allogeneic lymphocyte proliferation than transfected DCs with the vector plasmid pAS and untreated DCs at all dilutions. The most efficient shRNA plasmid vector against B7-DC was then used to silence the expression of B7-DC on DCs, the gene-modified DCs were pulsed with HBV-specific peptides, and HBV transgenic mice were immunized. After three rounds of immunization, the splenocytes were stimulated in vitro and tested for cytotoxicitic T lymphocyte activity, while the sera were used to detect the level of HBsAg and HBV DNA. The data demonstrated that blockade of B7-DC on DCs augmented the cytolytic activity induced by immunization with peptide-pulsed DCs and significantly reduced the concentration of serum HBsAg and HBV DNA, suggesting that silencing of B7-DC is of potential value in DC-based therapy of HBV infection.