A novel multi-component protein vaccine ECP001 containing a protein polypeptide antigen nPstS1 riching in T-cell epitopes showed good immunogenicity and protection in mice.

Tuberculosis (TB) is an infectious disease that seriously affects human health. Until now, the only anti-TB vaccine approved for use is the live attenuated Mycobacterium bovis (M. bovis) vaccine - BCG vaccine, but its protective efficacy is relatively low and does not provide satisfactory protection against TB in adults. Therefore, there is an urgent need for more effective vaccines to reduce the global TB epidemic. In this study, ESAT-6, CFP-10, two antigens full-length and the T-cell epitope polypeptide antigen of PstS1, named nPstS1, were selected to form one multi-component protein antigens, named ECP001, which include two types, one is a mixed protein antigen named ECP001m, the other is a fusion expression protein antigen named ECP001f, as candidates for protein subunit vaccines. were prepared by constructing one novel subunit vaccine by mixing or fusing the three proteins and combining them with aluminum hydroxide adjuvant, and the immunogenicity and protective properties of the vaccine was evaluated in mice. The results showed that ECP001 stimulated mice to produce high titre levels of IgG, IgG1 and IgG2a antibodies; meanwhile, high levels of IFN-γ and a broad range of specific cytokines were secreted by mouse splenocytes; in addition, ECP001 inhibited the proliferation of Mycobacterium tuberculosis in vitro with a capacity comparable to that of BCG. It can be concluded that ECP001 is a novel effective multicomponent subunit vaccine candidate with potential as BCG Initial Immunisation-ECP001 Booster Immunisation or therapeutic vaccine for M. tuberculosis infection.

Genetic Characterization Conferred Co-Resistance to Isoniazid and Ethionamide in Mycobacterium tuberculosis Isolates from Southern Xinjiang, China.

Background: Ethionamide (ETH), a structural analogue of isoniazid (INH), is used for treating multidrug-resistant tuberculosis (MDR-TB). Due to the common target InhA, INH and ETH showed cross-resistance in M. tuberculosis. This study aimed to explore the INH and ETH resistant profiles and genetic mutations conferring independent INH- or ETH-resistance and INH-ETH cross-resistance in M. tuberculosis circulating in south of Xinjiang, China. Methods: From Sep 2017 to Dec 2018, 312 isolates were included using drug susceptibility testing (DST), spoligotyping, and whole genome sequencing (WGS) to analyze the resistance characteristics for INH and/or ETH. Results: Among the 312 isolates, 185 (58.3%) and 127 (40.7%) belonged to the Beijing family and non-Beijing family, respectively; 90 (28.9%) were INH-resistant (INHR) with mutation rates of 74.4% in katG, 13.3% in inhA and its promoter, 11.1% in ahpC and its upstream region, 2.2% in ndh, 0.0% in mshA, whilst 34 (10.9%) were ETH-resistant (ETHR) with mutation rates of 38.2% in ethA, 26.2% in inhA and its promoter, and 5.9% in ndh, 0.0% in ethR or mshA; and 25 (8.0%) were INH-ETH co-resistant (INHRETHR) with mutation rates of 40.0% in inhA and its promoter, and 8% in ndh. katG mutants tended to display high-level resistant to INH; and more inhA and its promoter mutants showed low-level of INH and ETH resistance. The optimal gene combinations by WGS for the prediction of INHR, ETHR, and INHRETHR were, respectively, katG+inhA and its promoter (sensitivity: 81.11%, specificity: 90.54%), ethA+inhA and its promoter+ndh (sensitivity: 61.76%, specificity: 76.62%), and inhA and its promoter+ndh (sensitivity: 48.00%, specificity: 97.65%). Conclusion: This study revealed the high diversity of genetic mutations conferring INH and/or ETH resistance among M. tuberculosis isolates, which would facilitate the study on INHR and/or ETHR mechanisms and provide clues for choosing ETH for MDR treatment and molecular DST methods in south of Xinjiang, China.

Association of Single-Nucleotide Polymorphisms in the VDR Gene with Tuberculosis and Infection of Beijing Genotype Mycobacterium tuberculosis.

Background: The aim of the present study was to investigate the association between vitamin D receptor (VDR) gene polymorphism and tuberculosis susceptibility, as well as the potential interaction of host genetic factors with the heterogeneity of Mycobacterium tuberculosis in the population from Xinjiang, China. Methods: From January 2019 to January 2020, we enrolled 221 tuberculosis patients as the case group and 363 staff with no clinical symptoms as the control group from four designated tuberculosis hospitals in southern Xinjiang, China. The polymorphisms of Fok I, Taq I, Apa I, Bsm I, rs3847987 and rs739837 in the VDR were detected by sequencing. M. tuberculosis isolates were collected from the case group and identified as Beijing or non-Beijing lineage by multiplex PCR. Propensity score (PS), univariate analysis and multivariable logistic regression models were used to perform the analysis. Results: Our results showed that the allele and genotype frequencies of Fok I, Taq I, Apa I, Bsm I, rs3847987 and rs739837 in VDR were not correlated with tuberculosis susceptibility or lineages of M. tuberculosis. Two out of six loci of the VDR gene formed one haplotype block, and none of the haplotypes was found to correlate with tuberculosis susceptibility or lineages of M. tuberculosis infected. Conclusion: Polymorphisms in the VDR gene may not indicate susceptibility to tuberculosis. There was also no evidence on the interaction between the VDR gene of host and the lineages of M. tuberculosis in the population from Xinjiang, China. Further studies are nonetheless required to prove our conclusions.

Cloning, Expression, Invasion, and Immunological Reactivity of a Mammalian Cell Entry Protein Encoded by the mce1 Operon of Nocardia farcinica.

Bacterial mammalian cell entry (Mce) proteins have been implicated in pathogen invasion of mammalian host cells. The aim of this study was to examine the invasion-conferring ability of mce1E operon-encoded proteins, in vivo expression of Mce1E in cells from infected mice and rabbits, and Mce1E immunogenicity. Nocardia farcinica mce1E was cloned into pet30a(+) vectors, expressed in Escherichia coli, and purified. Invasion assays, transmission electron microscopy (TEM), immunoblots, and enzyme-linked immunosorbent assay (ELISA) detection of cytokines were conducted. TEM confirmed the invasion of HeLa cells by Mce1E-coated beads. The antigenicity of E. coli-expressed recombinant Mce1E was confirmed in immunoblots with sera from N. farcinica-infected mouse and rabbit sera. Co-incubation of Mce1E with splenocytes of N. farcinica-infected mice demonstrated upregulation of interferon (IFN-γ), but not interleukin (IL)-4 or IL-10, in the cultural supernatant. These findings demonstrate that Mce1E may facilitate N. farcinica interactions with and invasion of mammalian cells. Notably, Mce1E are expressed and elicited antibody responses in mice and rabbits during infection. Besides, it may play a role in cell-mediated immune reactions and cause host inflammation responses to N. farcinica infection.

[Under-5-mortality rate and causes of death in China, 2000 to 2010].

OBJECTIVE: To assess the changes and the leading cause of deaths for children under 5 years old, in China, during 2000 - 2010, with the aim of evaluation on the progress in achieving the relative goal set by "National Program of Action for Child Development in China (2001 - 2010)", and understanding the related challenges. METHODS: Data used in this study were collected from the population-based National Maternal and Child's Health Surveillance Network of China. Infant Mortality Rate (IMR), Under-5-mortality rate (U5MR) and the leading cause of deaths for under-5 children were analyzed. RESULTS: Nationwide IMR and U5MR in 2010 dropped by 59.3% and 58.7% respectively, compared to that in 2000. Decreases by 50.8% and 47.1% in IMR and U5MR were observed in urban areas, and 56.5% and 56.0% in rural areas during this period. Compared with data from 2000, the leading causes-specific U5MR in 2010 had significantly declined. The top 5 leading causes of death in 2010 were premature birth/low birth weight, pneumonia, birth asphyxia, congenital heart disease and accidental suffocation, but were different in urban and rural areas. In 2010, both IMR and U5MR from the rural areas were 2.8-folds than that of the urban areas. In addition, IMRs in the Middle and Western parts of China were 1.5 and 2.3-folds respectively of that in the East, and U5MR in Middle and West was 1.5 and 2.2-folds respectively of that in East. CONCLUSION: IMR, U5MR and the leading causes specific mortality rate in China declined remarkably from 2000 to 2010, and the goal set by "National Program of Action for Child Development in China (2001 - 2010)" had been successfully achieved. However, the disparity on child's health in regions and in urban or rural areas, still remained a challenge.

[Detecting overexpression level of HER2 gene in NSCLC by real-time quantitative PCR and the 2[-Delta Delta C(T)] method].

BACKGROUND AND OBJECTIVE: The evaluation of the expression level of the HER2 gene for the diagnosis and treatment of tumors is usually conducted using immunohistochemical techniques. The aim of the current study is to explore the feasibility of real-time quantitative PCR and the 2[-Delta Delta C(T)] method in detecting the level of HER2 gene overexpression in non-small cell lung cancer (NSCLC). METHODS: Real-time quantitative PCR and the 2[-Delta Delta C(T)] method were used to detect the level of HER2 gene overexpression in 212 lung cancer and matched non-tumor tissue specimens. RESULTS: The expression level of HER2 gene in lung cancer tissue was higher than that in the matched non-tumor tissue, with an overexpression rate of 34%. CONCLUSIONS: Real-time quantitative PCR and the 2[-Delta Delta C(T)] method can be used to detect the level of HER2 gene overexpression in NSCLC.

[Incidence and its trends on gastroschisis in some parts of China, 1996 - 2007].

OBJECTIVE: To investigate the incidence and its secular trends of gastroschisis in Chinese perinatal infants. METHODS: Data on perinatal infants was collected at hospitals under surveillance program in Chinese Birth Defects Monitoring Network from 1996 to 2007. Data on incidence, trend and related factors of gastroschisis in perinatal infants were carried out. Both χ(2) test and Poisson regression model were used to test the differences between residential areas, sex and maternal age. Both χ(2) trends test and Poisson regression model were applied to analyze the trends. RESULTS: A total of 6 308 594 perinatal infants were monitored during 1996 - 2007, including 1601 infants with gastroschisis to show the incidence as 2.54 per 10 000 births. The overall prevalence of gastroschisis in China did not change remarkably during the period of our research. The incidence rates of gastroschisis were significantly different between urban and rural areas, between different sex and different maternal age groups. The incidence of gastroschisis was lower in urban area than in rural area (RR = 0.58) and lower in female fetuses than in male fetuses (RR = 0.76), highest in the group younger than 20 years of age, which was 11.43 times than incidence of the 30 - 34 age group (RR = 11.432). CONCLUSION: The overall prevalence of gastroschisis in China did not show remarkable change during 1996 - 2007 but the incidence of gastroschisis a bit increased in the area of study and significant differences were seen in different sex, regions and maternal age groups. Mothers aged younger than 20 years old appeared to be a significant risk factor for the occurrence of gastroschisis.

Assessing the trend of gastroschisis prevalence in China from 1996 to 2007 using two analytical methods.

BACKGROUND: In recent years, the prevalence of gastroschisis has increased remarkably in some areas and remained unchanged in other areas; however, in general, there is a recent increasing trend compared to the 1970s and 1980s. In this study, we explored the time trend of gastroschisis prevalence in China during 1996 to 2007. METHODS: Data were retrieved from the hospital-based national monitoring database maintained by the Chinese Birth Defects Monitoring Network (CBDMN). The monitored subjects were infants born on the 28th gestational week or later, including live births and stillbirths. The maximal time for the diagnosis of a congenital malformation was the seventh day after birth. The secular trends on the overall prevalence and the different feature-specific prevalence of gastroschisis in China were analyzed using the linear chi-square test and the Poisson regression model. RESULTS: The overall prevalence of gastroschisis in China was 2.54 per 10,000 births during 1996 to 2007. Except for the prevalence of gastroschisis that significantly increased among infants whose mothers were 20 to 24 years old (p=0.0498 for the linear chi-square test, p=0.0032 for the Poisson regression model analysis) and significantly decreased among infants whose mothers were 30 to 34 years old (p=0.0177 for the Poisson regression model analysis), no significant changes were found in the overall and remaining feature-specific prevalences. CONCLUSION: The overall prevalence of gastroschisis in China did not change remarkably during 1996 to 2007; but the prevalence of gastroschisis significantly increased among infants whose mothers were 20 to 24 years old and decreased among infants whose mothers were 30 to 34 years old.