1.Investigation on Clonorchis sinensis infection and its risk factors in Futian District,Shenzhen City
Suyun FAN ; Xianghui SHI ; Jifei NIU ; Zhiping LIN ; Lilian LI
Chinese Journal of Schistosomiasis Control 2014;(6):699-700
Objective To understand the status of Clonorchis sinensis infection and its risk factors in Futian District Shen?zhen City so as to provide the evidence for formulating the strategy of prevention and control. Methods Two monitoring points were randomly selected and the permanent population aged≥3 years were investigated and their stool samples were detected for the eggs of C. sinensis by using Kato?Katz technique. Meanwhile the risk factors of infection were also investigated. Results A total of 743 subjects were examined and 7 persons were infected with C. sinensis and the infection rate was 0.94%. Most of the infected cases were focused on 20?40 and 40?60 age groups. The risk factors were eating raw or semi?cooked fish and the protec?tive factor was the use of separated cutting board for cooked or uncooked food. Conclusion The infection rate of C. sinensis is low in the population of Futian District. However the health education especially for diet health education still should be strengthened.
2.Association between CACNB2 gene polymorphisms and essential hypertension.
Qian SUN ; Xin WANG ; Ying HUANG ; Yunliang HU ; Jifei TANG ; Yan LIN ; Yuxin NIU ; Xiaoou WANG ; Bing DU
Chinese Journal of Medical Genetics 2013;30(3):340-344
OBJECTIVETo assess the association between single nucleotide polymorphisms (SNPs) of calcium channel β 2 subunit (CACNB2) gene and essential hypertension (EH) in ethnic Han Chinese in Wenzhou area, and to study the influence of rs7069292 alleles on gene expression with luciferase reporter technique.
METHODSSixty hundred and thirty seven Han Chinese with EH and 600 normal controls were enrolled. Genotypes of 6 SNP within CACNB2 gene including rs2228645, rs2357928, rs7069292, rs7099380, rs10764319 and rs11014166 were determined with matrix assisted laser desorption ionization/time of flight mass spectrometer (MALDI-TOF MS). A luciferase reporter gene plasmid containing the fragment flanking rs7069292 (-2831 bp to -2460 bp) in the 5' regulatory region of CACNB2 was constructed.
RESULTSCompared with the control, CT and TT genotypes for the rs7069292 locus were significantly more common in EH group (5.20% vs. 2.17%, 2.59% vs. 1.08%, P< 0.05). CC genotype was not found. Promoter activity for allele C of the rs7069292 locus was significantly increased compared with allele T (P< 0.05). No significant difference was detected for other 5 SNPs in terms of genotypes and allele frequency.
CONCLUSIONThe rs7069292 CT polymorphism of the CACNB2 gene is associated with EH in ethnic Han Chinese from Wenzhou area. A T>C mutation may affect the expression of CACNB2.
Aged ; Alleles ; Base Sequence ; Calcium Channels, L-Type ; genetics ; Case-Control Studies ; Cell Line ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide
3.False-positive HIV-1 nucleic acid testing results in patients with severe thalassemia after receiving cell and gene therapy
Yifan ZHONG ; Jifei NIU ; Yue LI ; Jing LIU ; Xiaohui WANG ; Hao LI ; Yongxia GAN ; Guilian LI ; Chenli ZHENG ; Chenglong LI ; Yifan CAI ; Zijie YANG ; Wei TAN ; Xiaozhen CHEN ; Tiejian FENG ; Cong JIN ; Jin ZHAO
Chinese Journal of Laboratory Medicine 2024;47(4):451-454
A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.
4.Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies.
Shenli ZHANG ; Qianru XU ; Jifei YANG ; Qingmei LI ; Yaning SUN ; Xueyang LI ; Yanan WANG ; Xiangxiang NIU ; Xiaotian QU ; Jinxuan CHEN ; Erqin ZHANG ; Gaiping ZHANG
Chinese Journal of Biotechnology 2022;38(5):1981-1993
The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
Animals
;
Antibodies, Viral
;
Chickens
;
HN Protein/metabolism*
;
Newcastle Disease/prevention & control*
;
Newcastle disease virus/metabolism*
;
Oryza/genetics*
5.The E248R protein of African swine fever virus inhibits the cGAS-STING-mediated innate immunity.
Yinguang LIU ; Wenping YANG ; Yuan WEN ; Qingli NIU ; Jifei YANG ; Guiquan GUAN ; Hong YIN ; Haixue ZHENG ; Dan LI ; Zhijie LIU
Chinese Journal of Biotechnology 2022;38(5):1837-1846
We researched the mechanism of African swine fever virus (ASFV) protein E248R in regulating the cGAS-STING pathway. First, we verified via the dual-luciferase reporter assay system that E248R protein inhibited the secretion of IFN-β induced by cGAS-STING or HT-DNA in a dose-dependent manner. The relative quantitative PCR analysis indicated that the overexpression of E248R inhibited HT-DNA-induced transcription of IFN-b1, RANTES, IL-6, and TNF-α in PK-15 cells. Next, we found that E248R interacted with STING by co-immunoprecipitation assay and laser confocal microscopy. Finally, we demonstrated that E248R inhibited the expression of STING protein by using Western blotting. We demonstrated for the first time that the E248R protein of ASFV suppressed the host innate immune response via inhibiting STING expression. The results are pivotal in extending the understanding of the ASFV immune escape and can guide the design of vaccines against ASFV.
African Swine Fever Virus/genetics*
;
Animals
;
DNA
;
Immunity, Innate
;
Nucleotidyltransferases/metabolism*
;
Signal Transduction
;
Swine