1.Screening of Proteins Binding to HBsAg from Human Pancreas cDNA Library by Yeast Two-hybrid System
Xiaochun WANG ; Jinqian ZHANG ; Chenyu ZHANG
Journal of Environment and Health 1992;0(05):-
Objective To screen proteins in pancreas cDNA library interacting with HBsAg protein.Methods After being amplified and identified,the human pancreas cDNA library plasmid was transformed into yeast Y187.Thereafter,the bait plasmid(PGBKT7-HBsAg) was transformed into yeast cells AH109.Then,two different transformed yeast cells were mated.The diploid yeast cells were plated on synthetic dropout nutrient medium(SD/-Trp/-Leu/-His/-Ade) and that medium contained X-?-gal for selection two times.After plasmids of true positive blue colonies were extracted and transformed to E.coli,the results were analyzed by DNA sequencing and blast searching in GenBank database.Results Six proteins binding to HBsAg were screened,including aconitase(ACO1),CDK5RAP3,carboxypeptidase,etc.Conclusion Most of these proteins play significant roles in the evolution of 2 type diabetes mellitus,which may supply some new clues for exploring the pathogenesis of abnormality of lipid metabolism and tglycometabolism in the patients with chronic hepatitis virus infection.
2.Ultrasonic osteotomy in mandibular angle reduction
Wang LIU ; Jinqian XU ; Qiaofang GAO ; Wenxian ZHANG ; Jun GUO
Chinese Journal of Medical Aesthetics and Cosmetology 2014;20(3):165-167
Objective To explore the clinical method and effect of ultrasonic osteotomy in mandibular angle reduction.Methods Under intronasal approach and general anesthesia,30 patients (5 male patients and 25 female patients) underwent intraoral prominent mandibular angle reductions with ultrasonic osteotomy.The jaw bone under periosteum was explored and the mental nerves protected.According to the clinical features of mandibular angle,curved mandibular angle osteotomy,mandible margin osteotomy or outer cotex splitting osteotomy were performed.Results All the osteotomy of prominent mandibular angle reductions could be completed with ultrasonic osteotomy.There were no complications of bleeding,nerve injury and malfracture happened.After 1 year following-up,the outlines of lower mandible were natural and concordant.Compared with traditional osteotomy devices,ultrasonic osteotomy was smoother and more precisely.Conclusions Ultrasonic osteotomy can improve the precise and safty of mandibular angle reduction.but compared with traditional osteotomy devices,it takes more time for osteotomy procedure.Surgeons need adapt to the different feeling and control methods.
3.Cloning and expression of genotype B and C hepatitis B virus in eukaryotic cells
Xiaoguang LI ; Yuan HONG ; Qi WANG ; Jinqian ZHANG ; Jun CHENG
Chinese Journal of Infectious Diseases 2010;28(1):10-13
Objective To construct recombinant full length genotype B and C hepatitis B virus (HBV)and to examine HBV DNA replication and hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)expressions in Huh7 cells. Methods The full length genotype B and C HBV DNA were extracted and amplified from two HBV infected patients. The recombinant plasmids were constructed by inserting the amplified HBV fragments into the eukaryotic expression vector,pHY106,which were then transfected into Huh7 cells. The cells transfected with blank pHY106 vector were used as control. HBV DNA replication at 72 hours of transfection was detected by Southern blot. The HBV DNA levels in Huh7 cells at 24,48,72,96 and 120 hours of transfection were determined by real-time polymerase chain reaction(PCR).Meanwhile, the HBsAg and HBeAg expression levels in the supernatants at 24,48,72,96 and 120 hours were determined by enzyme linked immunosorbent assay(ELISA).Results The recombinant plasmids expressing genotype B or C HBV DNA were successfully constructed.The HBV replicative intermediates in HBV core particles,including rcDNA dsDNA and ssDNA,were detected by Southern blot.HBV DNA level could reach 8 lg copy/mL which was by real-time PCR. HBsAg and HBeAg levels determined by ELISA peaked at 72 hours after transfection and then declined gradually. Conclusions The recombinant plasmids inserted with genotype B or C HBV DNA are constructed successfully, which can express high levels of HBsAg and HBeAg in Huh7 cells. This system provides a platform for studying the pathogenesis of B and C genotype HBV, the interaction between HBV and host, as well as exploiting new drugs against HBV.
4.Screening of proteins binding to hepatitis C virus E1 protein from human pancreas cDNA library
Xiaochun WANG ; Jinqian ZHANG ; Shuling WU ; Chenyu ZHANG ; Chaopin LI ; Guoli LI ; Qi WANG ; Jun CHENG
Chinese Journal of Infectious Diseases 2008;26(11):647-650
Objective To screen proteins from human pancreas cDNA library,which interact with hepatitis C virus(HCV)E1 protein.Methods The human pancreas cDNA library was amplified,purified and evaluated,and then the purified library plasmids were transformed into yeast strain Y187.The reconstructed plasmid pGBKT7-E1 was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp.The transformed AH109 mated with Y187 that contained the library plasmids.The diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-α-gal for selecting.The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5α.The plasmids in DH5α were extracted,sequenced and blasted.Result Sixteen proteins interacting with HCV E1 were found.Conclusion Some of the sixteen pancreatic proteins may be related with metabolisms of glucose and lipid.
5.Expression of Sirt1 in tissues of patients with gastric carcinoma
Wenxiang LIU ; Tao WANG ; Zhiqiang WU ; Jian YIN ; Jinqian ZHANG ; Peng HOU
Journal of Chinese Physician 2012;14(4):433-436
ObjectiveTo explore the changes in gene expression in tissues of patients with gastric carcinoma by cDNA microarray.MethodsA 44,000 gene cDNA microarray (Affymetfix) was used to examine gene expression in the tissues of patients with gastric carcinoma.The Sirt1 genes were identified and subjected to realtime PCR analysis.Results The expression of Sirt1 in tissues of gastric carcinoma [(0.318 ±0.032) ~ (0.169 ±0.013 ) time] was extremely lower than in tissues of para- gastric carcinoma ( 1 time).The results of Real-time PCR were consistent with those of genechip analysis.ConclusionsThe expression of Sirt1 was obviously different between tissues of gastric carcinoma and para - gastric carcinoma.Sirt1 may be associated with carcinogenesis of HCC.
6.Mutual interaction between hepatitis C virus core protein and apolipoprotein A Ⅰ
Jing SHEN ; Jun CHENG ; Chaopin LI ; Xiaoquan LI ; Qi WANG ; Guoli LI ; Jinqian ZHANG
Chinese Journal of Microbiology and Immunology 2009;29(8):697-700
group pACT-apoA Ⅰ and pBIND-core was higher than that in the negative control.Conclusion HCV core protein and apoA Ⅰ can interact in vivo.This study provides new theory for the mechanism of chronicity hepatitis C in fatty liver.
7.Role of occult hepatitis B virus infection in the pathogenesis of mul-tiple myeloma
Qixia WANG ; Qingren NI ; Jinqian DAI ; Liang GUO ; Jia XIE ; Jingjing REN
Chinese Journal of Clinical Oncology 2015;(14):700-704
Objective:To explore the relationship between hepatitis B virus (HBV) infection and the pathogenesis of multiple my-eloma (MM), in order to provide an epidemiological evidence for the prevention and treatment of MM. Methods:Clinical and epidemi-ological data of 185 MM patients and 182 non-tumorous patients were collected. Subjects were randomly selected from in-patients who were homeochronously admitted to the same five grade-III A hospitals, including Xi'an Central Hospital, Shaanxi People's Hospital, Xi'an Jiaotong University Xibei Hospital, and so on. MM patients were selected in terms of age and gender. Peripheral blood HBsAg was assayed by ELISA. If HBsAg was negative, the S and C-gene fragments of HBV DNA were tested using nested PCR . Fisher's ex-act test orχ2 test (SPSS statistical software) was used to compare the differences between the groups. Logistic regression was employed to examine the association between the pathogenesis of MM and HBV infection. Results:In MM patients, the HBsAg positive rate was 8.11% (15/185), the occult HBV infection (OBI) positive rate was 3.53% (6/170), and the total HBV infection rate was 11.35% (21/185). For the control group, the HBsAg positive rate was 4.40%(8/182), the OBI positive rate was 0.57%(1/174), and the total HBV in-fection rate was 4.95%(9/182). No statistical difference in HBsAg or OBI positive rate was found between the two groups (P>0.05). However, MM patients showed significantly higher total HBV infection rate than that of the controls [χ2=5.02, P<0.05;OR was 2.46 (95%CI:1.10-5.53, P<0.05)]. Additionally, the proportion of ISS stage III was significantly higher in MM patients with HBV infection than in uninfected MM patients (85.71%vs. 60.37%,χ2=5.15, P<0.05). Patients with HBV infection showed reduced albumin level (χ2=5.60, P<0.05) and aκ/λlight chain ratio (P<0.05) compared with uninfected patients. Conclusion:The risk of MM pathogenesis after HBV infection is significantly increased as OBI is included in the analyses. Furthermore, MM patients with HBV infection will develop more severe liver damage, indicating that OBI in MM patients with negative HBsAg should be screened before chemotherapy to pre-vent HBV reactivation.
8.Discussion on the revision of the general chapter for lotions in the Chinese Pharmacopoeia 2020 Volume Ⅳ
Jiana OU ; Shujuan LI ; Mei YANG ; Liangyong WU ; Weiling LI ; Wenxue ZENG ; Jinqian WANG ; Yuesheng WANG
Drug Standards of China 2024;25(5):443-445
Objective:To analyze the general chapter for lotions 0127 of the Chinese Pharmacopoeia 2020 Vol-ume Ⅳ,and discuss how to improve the general technical requirements of lotions 0127 in the Chinese Pharma-copoeia.Methods:By comparing the general chapter for lotions in domestic and international pharmacopoe-ias,the definition,classification,process,storage and corresponding inspection requirements were analyzed.Results and Conclusions:The general chapter for lotions 0127 of the Chinese Pharmacopoeia should be revised,including improvement of the definition,increasement of forms of preparations,and expansion of included varieties,so as to promote scientific regulation for drugs and exhibit a guiding role of the Chinese pharmacopoeia in drug control.
9.The effects of group B streptococcus colonization during late pregnancy on vaginal microbiota and neonatal outcomes
Yue HAN ; Jinqian YU ; Yajuan WANG ; Qi LIAO
Chinese Journal of Neonatology 2024;39(4):199-203
Objective:To study the effects of group B streptococcus (GBS) colonization during late pregnancy on vaginal microbiota and neonatal outcomes.Methods:From September 2020 to September 2021, pregnant women receiving prenatal care and delivered in our hospital were prospectively enrolled. They were assigned into GBS(+) group and GBS(-) group based on the results of GBS culture and/or PCR tests of vaginal secretions. The mothers were also assigned into early-onset infection(EO) group and non-early-onset infection(non-EO) group based on the presence or absence of early-onset infection of their neonates. The vaginal microbiota and neonatal outcomes were compared between these groups.Results:A total of 125 cases were enrolled, including 65(52.0%) in GBS(+) group and 60(48.0%) in GBS(-) group. 24 cases (19.2%) were in EO group and 101 cases (80.8%) in non-EO group. The incidences of premature rupture of membranes (PROM), amniotic fluid contamination, chorioamnionitis and early-onset neonatal infection in GBS(+) group were significantly higher than GBS(-) group(all P<0.05).The abundances of Streptococcus and Ureaplasma in vaginal flora of GBS(+) group were higher than GBS(-) group ( P<0.01), whereas Rhodococcus, Phyllobacterium and Bifidobacterium were lower than GBS(-) group ( P<0.05).The EO group had significantly higher abundance of enterococcus than the non-EO group ( P<0.05). Mothers with GBS colonization and neonates with early-onset infection had the highest abundance of Escherichia/Shigella ( P=0.04). Mothers with GBS colonization and neonates without early-onset infection showed the highest abundance of Gardnerella ( P=0.04). Conclusions:GBS colonization during late pregnancy increases the incidences of PROM, amniotic fluid contamination, chorioamnionitis and early-onset neonatal infection. GBS colonization causes abnormal vaginal flora in pregnant women. The increases of Ureaplasma, Streptococcus, Escherichia/Shigella and Enterococcus in vaginal microbiota maybe associated with early-onset neonatal infection.
10.Prognostic significance of the number of dissected lymph nodes in Siewert typeⅡadenocarcinoma of the esophagogastric junction without lymphatic metastasis
Jinqian WANG ; Dazhong WEI ; Mingran XIE ; Shibin XU ; Dongchun MA
Chinese Journal of Oncology 2016;38(4):300-304
Objective To investigate the correlation between the number of dissected lymph nodes ( LNs) and the prognosis of patients with node?negative Siewert type ⅡAEG. Methods 248 patients with Siewert type Ⅱ AEG treated in our hospital between January 1998 and December 2008 were retrospectively assessed. All cases underwent left transthoracic subtotal esophagogastrectomy with conventional two?field lymphadenectomy, and were histopathologically proved to be without lymph node involvement. The prognostic impact of the number of dissected LNs was analyzed. Results The overall median survival time and the 1?, 3?, and 5?year overall survival rates were 64 months, 80. 4%, 60. 8% and 51. 0%, respectively. Cox regression showed that the number of dissected LNs and the depth of tumor invasion were independent prognostic factors. Patients with a high number of negative LNs had better overall survival than patients with a low number of negative LNs ( P<0.05) . The patients had better long?term survival outcomes with more than 10 LN dissected for cases with pT1 tumor ( P<0.001) , and so did those with more than 15 LN dissected for cases with pT2?3 tumor (P=0.003, 0.018, respectively). Conclusion The number of negative lymph nodes and the depth of tumor invasion are independent prognostic factors for node?negative Siewert type ⅡAEG, and adequate lymph node dissection can improve the long?term survival.