1.Detection of human papilloma virus 16/18 by the quantitative fluorescent PCR
Shiping WANG ; Zhengjun JIA ; Zhuo HE ; Gan DAI ; Fei ZHONG
Chinese Journal of Microbiology and Immunology 2008;28(6):560-564
Objective To study the relation between the HPV6/18 virus infection and the development of pathological changes of cervix. Methods The number of HPV16/18 DNA copies and the expression rate of HPV16/18 E7 mRNA in the pathological cervix were examined by the quantitative fluorescent PCR combined with pathological diagnosis and immunohistochemistry staining. Results The HPV16 infection rates in chronic cervicitis group were much lower (7.4%) than that in the cervical intraepithelial neoplasia (CIN) groups and the cervical cancer group (69.6% and 72.7%), respectively. Statistical analysis showed that the difference of HPV16 DNA copies was not significant between the chronic cervicitis group and CIN groups. In contrast to the above mentioned result, the number of HPV DNA copies between the CIN groups and the cervical cancer group was significantly different. The HPV16 E7 gene expression rates in CIN Ⅰ, Ⅱ, Ⅲ and cervical cancer groups were 0,37.5%,42.9%,63.6%, respectively. Conclusion Ins more common than that with HPV18. The number of HPV16 DNA copies in cervical cancer tissues is markedly higher than that in CIN Ⅱ, Ⅲ groups. The HPV16 E7 mRNA expression is significantly increased in the cervical cancer, and it is more closely correlated to this pathological changes. The quantitative fluorescent PCR can be used to reflect the activity of HPV, and it is a useful method for the screening examination of HPV and for the early diagnosis and treatment of cervical caner.
2.Characteristics of peripheral blood CD45RA+and CD45RO+T lymphocyte subsets in patients with cGVHD and its clinical significance
Ying LOU ; Chunmiao WANG ; Ting LIN ; Yanfei LUO ; Huiquan GAN ; Maohua ZHUO
Chinese Journal of Immunology 2014;(8):1117-1120
To characterize the CD45RA+and CD45RO+T lymphocyte subsets in the peripheral blood of patients with cGVHD induced by allogeneic haematopoietic stem cell transplantation ( allo-HSCT ) and to explore their relations with the disease.Methods:The peripheral blood was collected from 64 patients after allo-HSCT,including 21 non-cGVHD patients,15 light grade cGVHD patients,18 mild grade cGVHD patients and 10 severe grade cGVHD patients,then CD4+CD45RA+,CD4+CD45RO+, CD8+CD45RA+and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry ( FCM).Results: Compared with the control,the percent of CD4+CD45RA+T lymphocyte in patients with light,mild and severe grade cGVHD decreased markedly (P<0.05),the percent of CD4+CD45RO+T lymphocyte increased markedly (P<0.05).But there were not obviously change in the patient with different grade cGVHD.The percent of CD8+CD45RA+,CD8+CD45RO+T lymphocyte did not change obviously.Conclusion:CD4+CD45RA+and CD4+CD45RO+may play an important role in the pathogenesis of cGVHD.
4.Cloning, Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain
Junlong YU ; Shiping WANG ; Zhuo HE ; Gan DAI ; Wenkai LI ; Xiaoxin JIANG ; Shaohua ZENG ; Xiaoqin XIAO ; Shaorui XU ; Zhiyue Lü ; Xianchu PENG ; Songhua ZHOU ; Xueqin LIU
Progress in Biochemistry and Biophysics 2006;33(7):665-672
A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the '3' and 5' ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt1 1 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase ofSchistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E. coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partia1 protection vaccine candidate.
5.Molecular diagnosis of thalassemia compound heterozygotes in PUMCH from 2012 to 2015
Zhuo YANG ; Bing HAN ; Yong GAN ; Yuqing PEI ; Yu CHEN ; Ali YE ; Qian CHEN ; Wei WU ; Xianyong JIANG ; Wei SU ; Jianhua HAN ; Wei CUI
Chinese Journal of Laboratory Medicine 2016;39(7):491-495
Objective To investigate the genotype distribution of thalassemia intermedia , major and compound thalassemia in Peking Union Medical College Hospital from 2012 to 2015. Methods Retrospectively 1 084 suspected thalassemia cases were analyzed in recent four years .Three common deletions of αglobin chain were detected by GAP-PCR.Three common point mutations of αglobin chain and seventeen common mutations of βglobin chain were identified by PCR reverse dot blot hybridization . Hemoglobin electrophoresis was carried out by Capillary Electrophoresis System .RBC associated parameters and morphology were analyzed by hematology analyzer and blood smear .Results 702 cases were confirmed to be thalassemia, and the positive rate was 64.76% (702 /1084).19 types of gene defects were detected. There were 4 types of gene defects in 23 case with α-thalassemia intermeida, including -α3.7 /--SEA , -α4.2 /--SEA , αCSα/--SEA and αQSα/--SEA , -α3.7 /--SEA to be the most common genotype (18 cases) .3 cases with β-thalassemia intermeida were confirmed and the genotypes were βCD 17(A→T) /β-29(A→G) , β-28(A→G) /β-28(A→G) andβIVS-Ⅱ-654(C→T) /βCD17(A→T) , respectively.There were also 1 βCD 41 -42(-TTCT) /βCD17(A→T) thalassemia major case. The genotypes of 2 HbE/β-thalassemia cases were βCD41 -42(-TTCT) /βE and βCD17(A→T) /βE.5 αβ-thalassemia including 2 βCD 41 -42(-TTCT) /βA compounded with αα/-α3.7 , 1βIVS-Ⅱ-654(C→T) /βA compounded with --SEA /αCSα, 1βCD17(A→T) /βA compounded with -α4.2 /ααand 1βCD 41 -42(-TTCT) /βA compounded with αCS α/αα.Rare and untypical haematological results were found , such as normal level HbA 2 and undetectable HbH, in compound heterozygosity with --SEA /αCS α and βIVS-Ⅱ-654(C→T) /βA. Conclusions The genotypes of thalassemia intermedia, major and compound thalassemia in Peking Union Medical College were highly variable .
6.Cloning,Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain
Junlong YU ; Shiping WANG ; Zhuo HE ; Gan DAI ; Wenkai LI ; Xiaoxin JIANG ; Shaohua ZENG ; Xiaoqin XIAO ; Shaorui XU ; Zhiyue L ; Xianchu PENG ; Songhua ZHOU ; Xueqin LIU
Progress in Biochemistry and Biophysics 2006;0(07):-
A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library ?gt11 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase of Schistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E.coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partial protection vaccine candidate.
7.Clinical analysis of 15 pediatric patients with tuberous sclerosis complex complicated by cardiac rhabdomyomas.
Guo-Qiang HUANG ; Qiong-Xiang ZHAI ; Jun-Hao YU ; Chun WANG ; Mu-Qing ZHUO ; Lin-Gan WANG
Chinese Journal of Contemporary Pediatrics 2015;17(5):477-481
OBJECTIVETo investigate the clinical features in children with tuberous sclerosis complex (TSC)-associated cardiac rhabdomyomas (CRM).
METHODSThe clinical data of 15 children with TSC complicated by CRM were collected. The clinical features of the patients were analyzed, and TSC gene mutations were detected.
RESULTSEleven cases (73%) developed multiple CRM. The majority of the tumors were located in the left and right ventricles. Most tumors presented as a round-like hyperechogenic mass with a clear margin on echocardiography. Arrhythmias occurred in 3 patients and 2 patients experienced heart failure. Gene mutation tests were performed in 2 patients, and pathogenic mutations were detected in both patients, which were TSC1 mutation and TSC2 mutation, respectively. Three patients were followed up for 6 to 38 months, and their CRM shrank or regressed spontaneously.
CONCLUSIONSTSC-associated CRM is generally multiple. Heart failure and arrhythmias may occur in some patients. Echocardiography is important for diagnosis of CRM. TSC-associated CRM has an inclination to spontaneous regression. TSC can be diagnosed at a molecular genetic level by TSC gene mutation detection.
Child, Preschool ; Female ; Heart Neoplasms ; complications ; genetics ; Hemodynamics ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Rhabdomyoma ; complications ; genetics ; Tuberous Sclerosis ; etiology ; Tumor Suppressor Proteins ; genetics
8.Clinical application of intraosseous internal fixation in ankle-foot arthrodesis
Zhuo CHEN ; Yaokai GAN ; Dingwei SHI ; Chen JIN ; Jie ZHAO ; Kerong DAI
Chinese Journal of Orthopaedic Trauma 2019;21(4):328-332
Objective To investigate the short- to medium-term therapeutic effects of a new internal fixation device,intraosseous internal fixation(IO-FIX),in the foot-ankle arthrodesis.Methods From August 2016 to December 2018,32 patients(40 feet) underwent foot-ankle arthrodesis with IO-FIX at Department of Orthopaedic Surgery,The Ninth Peopled Hospital of Shanghai.They were 6 males and 26 females,aged from 23 to 83 years(61.7±15.1 years).There were 28 cases(36 feet) of hallux valgus,2 cases(2 feet) of ankle arthritis,one case(1 foot) of ankle rheumatoid arthritis and one case(1 foot) of naviculocuneiform arthrosis.The therapeutic effects were assessed in terms of imaging evaluation,American Orthopaedic Foot and Ankle Society(AOFAS) score,fusion rate and full weight-bearing time.Results The follow-up ranged from 3.0 to 31.3 months(mean,12.6 months).The total fusion rate was 95.0%(38/40);the total AOFAS score increased significantly from preoperative 41.9±9.5 to postoperative 86.9±4.7(P<0.05).The 28 patients(36 feet) with hallux valgus achieved a successful fusion rate of 97.2%(35/36) and full weight-bearing at 8 weeks after operation.Their fusion time ranged from 45 to 64 days(57.1 days).Their metatarsophalangeal angle was decreased significantly from 52.10±13.50 preoperatively to 13.40±4.90 postoperatively and their 1-2 intermetatarsal angle significantly from 14.5°±2.4°to 8.90±2.4°(P<0.05).Their AOFAS ankle-hindfoot score was improved significantly from 41.8±9.9 to 87.0±4.8(P<0.05).All the 3 cases of ankle arthrodesis achieved good bone union and full weight-bearing within 12 weeks and their AOFAS ankle-hindfoot score was improved significantly from preoperative 43.4±3.5 to 86.3±3.5 at the final follow-up.Conclusion In foot-ankle joint arthrodesis,due to advantages of stable fixation,zero profile,limited soft tissue irritation,reinforced bone bridge and easy reproducibility,10-FIX can lead to satisfactory short- to mid-term therapeutic effects.
9.Pharmacokinetics of two recombinant humanized monoclonal antibodies against ricin in rhesus monkeys
Ya GAO ; Xiao-xia ZHU ; Zhi-yun MENG ; Hui GAN ; Ruo-lan GU ; Zhuo-na WU ; Wen-zhong SUN ; Gui-fang DOU
Acta Pharmaceutica Sinica 2022;57(2):480-483
Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between
10.Analysis of risk factors for contrast agent extravasation in coronary CT angiography
Haihong LONG ; Fei HAN ; Liangying GAN ; Zhuo LIU ; Jin CHENG ; Chao SUN ; Ling FU
China Medical Equipment 2024;21(9):97-101
Objective:To analyze the influencing factors of contrast agent extravasation in coronary CT angiography(CTA)examination,and to formulate intervention measures.Methods:A retrospective selection of data from 583 patients who underwent coronary CTA at Peking University People's Hospital from January to December 2023 was conducted.Logistic regression was used to analyze the patients'general information and injection protocols,and the risk factors of contrast agent extravasation were determined.Results:Among the 583 patients included,11 patients had contrast agent extravasation during CTA examination,with an extravasation rate of 1.887%.The contrast agent was all extravasated into the subcutaneous tissue,and the CT value did not reach the trigger criteria.Gender,education level,diabetes mellitus,history of intravenous chemotherapy,age,weight,body mass index(BMI),injection rate and injection dose were all associated with the occurrence of contrast agent extravasation,the difference was statistically significant(x2=18.911,7.563,16.567,4.279,t=3.576,3.244,1.865,4.297,6.532,P<0.05).Age,education level,history of intravenous chemotherapy,diabetes mellitus,injection rate and injection dose were risk factors for contrast agent extravasation in coronary CTA(OR=1.008,1.372,1.029,5.092,0.975,1.421,P<0.05).Conclusion:Factors such as low education level,advanced age,history of intravenous chemotherapy,high injection rate and large injection dose can increase the risk of contrast agent extravasation in coronary CTA examination.Radiology staff should closely monitor high-risk patients,strengthen monitoring of intravenous injection of contrast agents for coronary CTA examination,and reduce the occurrence of contrast agent extravasation.