1.Relationship between hepatitis B virus markers in umbilical cord blood and maternal serum
Youlin SHAO ; Suocai ZHANG ; Longgen LIU ; Dinggui CHEN ; Fengcai GUO
Chinese Journal of Clinical Infectious Diseases 2013;6(4):205-208
Objective To investigate the correlation of hepatitis B virus (HBV) markers between umbilical cord blood and maternal serum.Methods A total of 340 HBsAg positive mothers who delivered at the Third People' s Hospital of Changzhou during August 2009 and November 2010 were included in the study.HBV markers in the maternal serum before childbirth and umbilical cord blood after birth were quantitatively detected.The neonates received 3 doses of hepatitis B vaccine and 2 doses of hepatitis B immunoglobulin (HBIG),and followed up for 12 months.Measurement data were expressed as median (M),and Kruskal-Wallis test and Spearman correlation analysis were performed.Results There were 175HBeAg-positive and 165 HBeAg-negative mothers,and a total of 341 infants were delivered.The positive rates of HBsAg,anti-HBs,HBeAg,anti-HBe and anti-HBc in maternal serum were 100.00%,0.0%,51.47%,38.82% and 99.41%,respectively; while those in umbilical cord bloods were 14.66%,0.59%,26.69%,39.88% and 95.31%,respectively.HBsAg concentration in maternal serum of HBsAg-positive umbilical cord blood group was higher than that of HBsAg-negative umbilical cord blood group (419.40∶ 387.95,x2 =4.592,P < 0.05) ; while HBsAg concentration in umbilical cord blood of HBeAg-positive maternal serum was higher than that of the HBeAg-negative maternal serum group (0.04 ∶ 0.01,x2 =5.674,P < 0.05).Anti-HBe and anti-HBc in umbilical cord blood were positively correlated with those in maternal serum (r =0.838,0.764,P < 0.01).Seven out of 62 (11.29%) infants were infected with HBV in HBeAg-positive maternal serum group; while no infant infected in HBeAg-negative matemal serum group.Conclusion The higher maternal serum HBsAg concentration,the greater the risk of perinatal transmission,and infants born by HBeAg-positive mothers are of high risk of HBV infection.
2.Application of CRISPR/Cas9 genome editing technology for inhibition of hepatitis B virus replication
Tao WU ; Xiaojuan ZHU ; Lunbiao CUI ; Huan FAN ; Yin CHEN ; Xiling GUO ; Kangchen ZHAO ; Zhiyang SHI ; Fengcai ZHU
Chinese Journal of Microbiology and Immunology 2015;(8):600-605
Objective To evaluate the practicability of using CRISPR/Cas9 genome editing tech-nology for inhibition of hepatitis B virus ( HBV) replication. Methods Two sgRNA targeting sites were de-signed for the S region of HBV genome. The CRISPR/Cas9 expression plasmids specific for HBV were con-structed and then transfected into a cell line expressing HBV genome(HepG2-N10). The cytotoxicity of cells transfected with different expression plasmids were detected by MTT assay. The levels of hepatitis B surface antigen ( HBsAg ) were determined by using chemiluminescent immunoassay ( CLIA ) . The expression of HBV at mRNA level was analyzed by quantitative real-time PCR ( qRT-PCR) . The qPCR was performed for the detection of extracellular and intracellular HBV DNA. The next-generation sequencing ( NGS) Illumina MiSeq Platform was used to analyze HBV genome editing. Results No significant cytotoxic effects were de-tected in HepG2-N10 cells transfected with different expression plasmids. Compared with the cells carrying pCas-Guide-GFP-Scramble, the levels of HBsAg in the supernatants of transfected cell culture harboring pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were decreased by 24. 2% (P<0. 05) and 16. 9% (P>0. 05), respectively. The levels of HBsAg in cells transfected with pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were respectively decreased by 16. 4% (P>0. 05) and 32. 1% (P>0. 05) as compared with that of pCas-Guide-GFP-Scramble transfected group. The expression of HBV at mRNA level was inhibited as indica-ted by the results of qRT-PCR. Moreover, the levels of extracellular HBV DNA were respectively suppressed by 23% (P>0. 05) and 35% (P<0. 05), and the levels of intracellular HBV DNA were respectively sup-pressed by 7. 2% (P>0. 05) and 18% (P>0. 05). Different types of insertion/deletion mutation were de-tected in HBV genome by high-throughput sequencing. Conclusion HBV-specific CRISPR/Cas9 system could inhibit the expression of HBV gene and the replication of virus. Therefore, the CRISPR/Cas9 genome editing technology might be used as a potential tool for the treatment of persistent HBV infection.
3.Expressions of CD117 and CD28 in patients with newly diagnosed multiple myeloma and their clinical significances
Fangfang GE ; Wenliang TIAN ; Huimin SUN ; Fengcai GAO ; Hui SUN ; Ling SUN ; Rong GUO ; Dingming WAN ; Linxiang LIU ; Yanfang LIU ; Xinsheng XIE ; Zhongxing JIANG
Journal of Leukemia & Lymphoma 2019;28(5):263-267
Objective To investigate the expressions of CD28 and CD117 in patients with newly diagnosed multiple myeloma (MM) and their clinical significances. Methods The clinical data of 115 newly diagnosed MM patients in the First Affiliated Hospital of Zhengzhou University from May 2015 to December 2017 were retrospectively analyzed. The expressions of CD28 and CD117 were detected by using multiparameter flow cytometry. The relationship between the expressions of CD28 and CD117 and MM staging and clinical parameters was analyzed. The staging was performed according to the International Staging System (ISS). Results Among these 115 patients, there were 15 patients with CD117 positive and 30 patients with CD28 positive. Erythrocyte sedimentation rate (r = -0.481, P = 0.039), Cˉreactive protein level (r = -0.314, P=0.015), the proportion of plasma cells detected by bone marrow cytology (r=-0.027, P=0.001) were negatively correlated with CD117 positive expressions. CD28 positive expression was positively correlated with lactate dehydrogenase level (r = 0.249, P = 0.033) and ISS stage (r = 0.319, P = 0.017), while it was negatively correlated with hemoglobin level (r = -0.372, P = 0.026). CD28 positive was associated with light chain type, and nonˉsecretory type mostly occurred (P = 0.016). The incidence of osteolytic lesions in CD28 positive group and CD117 positive group was high, but there was no statistical difference between CD28 positive group, CD117 positive group and CD28 negative group, CD117 negative group (P = 0.052, P=0.479). Conclusions The positive expression of CD117 in the early stage of MM patients is higher than that in the advanced stage, and the expression of CD28 positive in the advanced stage of MM patients is higher than that in the early stage. CD28 and CD117 can be used as indicators of prognosis stratification in the patients with newly diagnosed MM.
4.Analysis of virulence genes and the chromosomal 16S rRNA gene polymorphism of Yersinia enterocolitic in Jiangsu Province,2015
Lu ZHOU ; Chen DONG ; Hui GUO ; Yinhua WU ; Yongjie ZHANG ; Jing AI ; Ling GU ; Changjun BAO ; Minghao ZHOU ; Fengcai ZHU ; Zhongming TAN
Chinese Journal of Infectious Diseases 2018;36(1):34-37
Objective To reveal the virulence genes and the polymorphisms of chromosomal 16S rRNA gene of Yersinia enterocolitic strains isolated from different districts in Jiangsu Province,2015. Methods Five virulence genes(ail,virF,yadA,ystA and ystB)of Yersinia enterocolitic strains isolated from different districts in Jiangsu Province were detected by using polymerase chain reaction(PCR),and phylogenetic analysis of chromosomal 16S rRNA gene was performed by amplification and sequencing. Results In this study,73 Yersinia enterocolitic strains were collected in Jiangsu Province in 2015.Among them,56(76.7%)strains carried virulence genes,and ail-virF-yadA -ystA -ystB+were the dominate types in diarrhea patients and other hosts.All strains can be clustering into 4 groups according to the phylogenetic analysis of chromosomal 16S rRNA gene.Conclusions The non-pathogenic Yersinia enterocolitic(ystB+)is the dominant strain in Jiangsu province,and the pathogenic strains are also found in this region.The result of phylogenetic analysis of chromosomal 16S rRNA gene and the profiles of virulence genes are highly consistent.
5. Relationship between liver controlled attenuation parameters and body fat mass and its distribution
Youlin SHAO ; Suocai ZHANG ; Jianming WU ; Fengcai GUO ; Longgen LIU ; Chunyan YE ; Tong YAN ; Qi CAO ; Fan ZHANG ; Jing WANG ; Yonghua MAO ; Jiangao FAN
Chinese Journal of Hepatology 2019;27(10):754-759
Objective:
To explore the relationship between liver controlled attenuation parameters (CAP) and body fat mass and its distribution.
Methods:
From May to December 2018, 978 adult patients visited at the fatty liver center of the Third People's Hospital of Changzhou were treated. The patient's liver controlled attenuation parameters were measured by transient elastography and the body fat mass and its distribution were measured by bioelectrical impedance technology. Pearson’s correlation coefficient was adopted to describe the correlation between liver CAP value and body mass index (BMI), body fat mass index (BFMI), trunk fat mass index (TFMI), limbs fat mass index (LFMI) and visceral fat area (VFA). Receiver operating characteristic curve (ROC) and area under the curve (AUC) were used to evaluate BMI, BFMI, TFMI, LFMI and VFA to differentiate the cut-off points and efficacy of CAP for diagnosing grading of fatty liver changes in S0-1 and S2-3.
Results:
In 653 cases of male, S0 ~ S3 accounted for 4.90%, 3.37%, 22.36% and 69.37%, respectively, and in 325 cases of females, S0 ~ S3 accounted for 7.38%, 6.46%, 13.23% and 72.92%, respectively. Female patients had more visceral, trunk and limbs fat than male (