1.Development of duplex TaqMan PCR assay for detection of specific gene sequence from Bacillus anthracis
Shi-Kui WANG ; Ji-Hong HU ; Ming HOU ; Cheng GONG ; Zi-Yu SHEN ; Hui GUO ; Jian-Ping CAI ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To establish a rapid assay with high sensitivity and specificity based on the sequences for group specific gene (GS) and pathogenicity island pag A gene.Methods The PCR primers and probes were designed after the whole sequence was systemically analyzed with bio-informafion tools and blasted with Genebank database.The amplicons were inserted into plasmids so that they could be used as the standard templates to evaluate the sensitivity of the diagnostic system.This assay was based on TaqMan probes and portable Smartcycle PCR machine.Results The detection level was approximately 100 copies per reaction.There was no cross-reaction with other species of Bacillus.This assay could be completed in one hour in laboratory.Conclusion The duplex TaqMan PCR assay could be used to detect Bacillus anthracis rapidly with high sensitivity and specificity.
2.The value of 18F-FDG PET/CT in the diagnosis of secondary malignant peripheral nerve lesion
Lei, FANG ; Shi-cun, WANG ; Bo, PAN ; Feng-lin, ZHAN ; Qiang, XIE ; Ji-kui, XIE ; Jiu-lin, LI ; Sai-juan, SHEN
Chinese Journal of Nuclear Medicine 2010;30(4):222-225
Objective To investigate the characteristics and diagnostic value of 18F-fluorodeoxyglucose (FDG) PET/CT in patients with secondary malignant peripheral nerve lesions. Methods 18F-FDG PET/CT studies of 8 cases of secondary malignant peripheral nerve lesions confirmed by histopathology or follow-up were analyzed retrospectively. The maximum standardized uptake value ( SUVmax ) of infiltrating peripheral nerves and contralateral normal peripheral nerves was measured and compared with their morphological appearances on CT. Paired student t-test was performed by SPSS 10.0. Results Twelve secondary malignant peripheral nerve lesions with high 18F-FDG metabolism were found in 8 cases. On PET imaging,the lesions distributed along the neurovascular tissues or intervertebral foramina with appearances resembling those of fibre bundles,radices or nodes on PET but no density differences with the surrounding soft tissue or fat planes on CT. The SUVmax was 6.86 ± 3.87. The contralateral normal peripheral nerves showed no abnormal 18F-FDG uptake with a SUVmax of 1.10 ±0.46,which was significantly different from that of the secondary malignant peripheral nerve lesions (t = 9.231,P < 0.001 ). Conclusion 18 F-FDG PET/CT may be useful in locating the secondary malignant peripheral nerve lesions and in assessing its regional infiltration.
3.Construction of the stable expression system of NIH3T3 fibroblast with pcDNA3.1(-)-hTGFbeta3 and the study on the proliferation of the system fibroblasts.
Shi-Jie TANG ; Si-Tian XIE ; Su-Luan HU ; Zhi-Qiang XIAO ; Ji-Kui SHEN ; Hong YI
Chinese Journal of Plastic Surgery 2006;22(2):109-112
OBJECTIVETo construct and identify the stable expression system of NIH3T3 fibroblast with eukaryotic expression vector of human transforming growth factor beta3 (pcDNA3.1 (-)/TGFbeta3). So as to investigate the proliferation of NIH3T3 fibroblasts transfected with hTGFbeta3 gene stably.
METHODSThe stable transfection of NIH3T3 fibroblasts with recombinant plasmid expressing hTGFbeta3 was established by using LipofectamineTM2000 and G418 selection. The mRNA and protein expression of TGFbeta3 were detected by the RT-PCR and Western blot method, respectively. Microscope and MTT were adopted to examine the proliferation of the stable expression system of fibroblasts with hTGFbeta3.
RESULTSAfter G418 selection, RT-PCR and Western blot analysis, 7 out of 10 cell lines transfected with pcDNA3.1 (-)/TGFbeta3 expressed with very high level of TGFbeta3, as compared with vector control transfectants that showed no expression, and compared with the other cell lines that expressed relatively low level. The stable transfection of NIH3T3 fibroblasts growth slowed down significantly (P < 0.05).
CONCLUSIONThe stable expression system of NIH3T3 fibroblast with hTGFbeta3 were constructed successfully. The TGFbeta3 gene could inhibit the proliferation of NIH3T3 fibroblasts in vitro.
Animals ; Cell Proliferation ; Fibroblasts ; metabolism ; Humans ; Mice ; NIH 3T3 Cells ; Plasmids ; Transfection ; Transforming Growth Factor beta3 ; genetics ; metabolism
4.Epidemiological characteristic of first case of locally identified A/H1N1 secondary cases caused by imported source of infection in China
Zhi-Cong YANG ; Tie-Gang LI ; Yu-Fei LIU ; Xin-Wei WU ; Jun YUAN ; Chao-Jun XIE ; Kui-Biao LI ; Li-Yun JIANG ; Yi-Yun CHEN ; Xin-Wu MAO ; Hai-Lin LI ; Liu-Hua ZHAN ; Xiao-Ling XIAO ; Jian-Ping LIU ; Ji-Chuan SHEN ; Wen-Feng CAI ; Ming WANG
Chinese Journal of Epidemiology 2009;30(7):684-686
Objective To study the first locally identifcd A/HINI secondary cases outbreak in China. Methods Interview and field investigation were integrated to describe the whole process of transmission on each case and to illustrate the relationships between the onset of the disease and the retated factors. Results Two contact persons appearanced fever and whose throat swabs were tested positive to H1N1 viral nucleic acid. The two had a history of contact in a short distance with the initial imported case without any protective measure in the poor air ventilation. The patients clinical situation was slight. The incubation was between 37 hours and 57 hours. No other new case was found after intervention as isolation and antisepsis were taken. Conclusion This event was proved to be an outbreak of local A/H1N1 secondary cases caused by the imported case. The main mode of transmission was personal contact in a short distance without protection, through air and droplet. The locus with poor air ventilation was high risk place. Contact persons should be observed seven days and tested continuously.Infectivity and pathogenicity of the A/H1N1 virus were limited and appeared weakened by generations. Patient's condition was related with persistence and frequency of contact with the infection sources. Enhancing management of contact persons, health education, early diagnose, early treatment and early insulation were effective measures of controling and prenventing the spread A/H1N1.
5.Effects of laparoscopic hyperthermic intraperitoneal perfusion chemotherapy combined with intraperitoneal and systemic chemotherapy treatment in patients with untreated gastric cancer with peritoneal metastasis.
Shen LI ; Kan XUE ; Hong Mei DAI ; Yin Kui WANG ; Fei SHAN ; Zi Yu LI ; Jia Fu JI
Chinese Journal of Gastrointestinal Surgery 2023;26(5):442-447
Objective: To investigate the efficacy of laparoscopic hyperthermic intraperitoneal perfusion chemotherapy combined with intraperitoneal and systemic chemotherapy (HIPEC-IP-IV) in the treatment of peritoneal metastases from gastric cancer (GCPM). Methods: This was a descriptive case series study. Indications for HIPEC-IP-IV treatment include: (1) pathologically confirmed gastric or esophagogastric junction adenocarcinoma; (2) age 20-85 years; (3) peritoneal metastases as the sole form of Stage IV disease, confirmed by computed tomography, laparoscopic exploration, ascites or peritoneal lavage fluid cytology; and (4) Eastern Cooperative Oncology Group performance status 0-1. Contraindications include: (1) routine blood tests, liver and renal function, and electrocardiogram showing no contraindications to chemotherapy; (2) no serious cardiopulmonary dysfunction; and (3) no intestinal obstruction or peritoneal adhesions. According to the above criteria, data of patients with GCPM who had undergone laparoscopic exploration and HIPEC from June 2015 to March 2021 in the Peking University Cancer Hospital Gastrointestinal Center were analyzed, after excluding those who had received antitumor medical or surgical treatment. Two weeks after laparoscopic exploration and HIPEC, the patients received intraperitoneal and systemic chemotherapy. They were evaluated every two to four cycles. Surgery was considered if the treatment was effective, as shown by achieving stable disease or a partial or complete response and negative cytology. The primary outcomes were surgical conversion rate, R0 resection rate, and overall survival. Results: Sixty-nine previously untreated patients with GCPM had undergone HIPEC-IP-IV, including 43 men and 26 women; with a median age of 59 (24-83) years. The median PCI was 10 (1-39). Thirteen patients (18.8%) underwent surgery after HIPEC-IP-IV, R0 being achieved in nine of them (13.0%). The median overall survival (OS) was 16.1 months. The median OS of patients with massive or moderate ascites and little or no ascites were 6.6 and 17.9 months, respectively (P<0.001). The median OS of patients who had undergone R0 surgery, non-R0 surgery, and no surgery were 32.8, 8.0, and 14.9 months, respectively (P=0.007). Conclusions: HIPEC-IP-IV is a feasible treatment protocol for GCPM. Patients with massive or moderate ascites have a poor prognosis. Candidates for surgery should be selected carefully from those in whom treatment has been effective and R0 should be aimed for.
Male
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Humans
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Female
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Middle Aged
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Aged
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Aged, 80 and over
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Young Adult
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Adult
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Stomach Neoplasms/surgery*
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Peritoneal Neoplasms/secondary*
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Hyperthermic Intraperitoneal Chemotherapy
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Percutaneous Coronary Intervention
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Hyperthermia, Induced/methods*
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Combined Modality Therapy
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Laparoscopy/methods*
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Antineoplastic Combined Chemotherapy Protocols/therapeutic use*
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Perfusion
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Cytoreduction Surgical Procedures
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Survival Rate