2.THE DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS IN ANIMAL TISSUE BY RT-PCR TECHNIQUE
Gao-Ming LOU ; Wei-Xian DU ; Ao-Bin YANG ; Xiu-Rong ZHOU ; Ming-Qian XIE ;
Microbiology 1992;0(04):-
A set of primers amplified the VP1 gene of foot-and-mouth disease vims (FMDV) was designed and synthesized. A reverse transcription-polymerase chain reaction (RT-PCR) technique detected the RNA of FMDV was established after selecting the best purification method, reagents and reaction conditions. Samples of fresh milk, lymph node, spinal cord, vesicular skin, milk powder, cotton swab, mouse and meat in daughter-house were detected by RT-PCR, positive rates were41.4% (24/58), 13.33% (2/15), 20% (1/5), 100% (1/1), 100% (1/1), 37.5% (12/32), 100% (2/2) and 10% - 70%, respectively. However, positive rate of cockroach detected by RT-PCR was 0. The results showed that the established FMDV RT-PCR technique provided a more sensitive, specific and reliable method for diagnosis and epizootic study of the foot-and-mouth disease.
3.Relationship between CCR5 gene polymorphism and condyloma acuminata.
Ya-Gang ZUO ; Bao-Xi WANG ; Xiu-Rong LIU ; Peng ZHAO ; Qian-Shu YAN
Acta Academiae Medicinae Sinicae 2008;30(1):95-97
OBJECTIVETo investigate the relationship between CCR5 delta32 gene polymorphism and condyloma acuminata.
METHODSWe used polymerase chain reaction to amplify the CCR5 gene fragments in 60 patients with condyloma acuminata and 50 age- and sampling date-matched controls, and compared the difference of genotypes between these two groups.
RESULTSNo genotype difference was found between these two groups.
CONCLUSIONCondyloma acuminata are not associated with genetic polymorphism of CCR5 delta32 gene.
Condylomata Acuminata ; genetics ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Receptors, CCR5 ; genetics
4.Uptake of 2-NBDG by human breast cancer cells in vitro.
Hui HU ; Xiu-hong SHAN ; Wei ZHU ; Hui QIAN ; Wen-rong XU ; Ya-fei WANG
Chinese Journal of Oncology 2010;32(7):507-510
OBJECTIVEThe purpose of this study was to assess the feasibility of fluorescent 2-deoxyglucose analog, 2-[N-(7-nitrobenz-2-oxa-1, 3-diaxol-4-yl)amino]-2-deoxyglucose (2-NBDG), that could be taken up by breast cancer cells highly expressing glucose transporter 1 (GLUT-1). The purpose of this study was to clarify if a fluorescent 2-deoxyglucose analog, 2-[N-(7-nitrobenz-2-oxa-1, 3-diaxol-4-yl)amino]-2-deoxyglucose (2-NBDG), can be taken up by breast cancer cells highly expressing glucose transporter 1 (GLUT-1), and to assess whether it can be used as a targeting imaging agent.
METHODSThe expressions of GLUT-1 mRNA and protein in breast cancer MDA-MB-231 cells were detected by RT-PCR and immunohistochemistry, respectively. The difference of GLUT-1 protein expression between breast cancer MDA-MB-231 cells and MCF-7 cells was compared by Western blot. Secondly, MDA-MB-231 cells which were grown in 6-well plates were incubated with 2-NBDG, and the result of 2-NBDG uptake was analyzed by fluorescence microscopy and flow cytometry. The difference of 2-NBDG absorption in MDA-MB-231 and MCF-7 cells was compared by flow cytometry.
RESULTSThe results of RT-PCR and immunohistochemistry confirmed that MDA-MB-231 cells highly expressed GLUT-1. Furthermore, Western blot revealed that GLUT-1 expression of MDA-MB-231 cells (0.946 ± 0.007) was higher than that in the MCF-7 cells (0.833 ± 0.010). Fluorescence microscopic and flow cytometric analysis showed that 2-NBDG was uptaken rapidly by MDA-MB-231 cells. Addition of 50 mmol/L D-glucose to the media with 2-NBDG reduced its uptake by 46.0%. Moreover, flow cytometry indicated that the fluorescence intensity of MDA-MB-231 cells (25.10 ± 0.57) was higher than that of MCF-7 cells (10.12 ± 0.62) when incubated with 2-NBDG for 20 minutes.
CONCLUSIONThe preliminary data clearly demonstrate that 2-NBDG is taken up and accumulated in breast cancer cells that highly express GLUT-1, and may be used as an optical probe for glucose uptake in hypermetabolic malignant cells.
4-Chloro-7-nitrobenzofurazan ; analogs & derivatives ; pharmacokinetics ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Deoxyglucose ; analogs & derivatives ; pharmacokinetics ; Female ; Flow Cytometry ; Glucose Transporter Type 1 ; genetics ; metabolism ; Humans ; Immunohistochemistry ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
5.Construction and characterization of the stk and stp1 knockout mutant in streptococcus suis type 2
Chao-Long LI ; Jian ZHANG ; Qian-Qian WU ; Hong-Fen HOU ; Hui-Fang ZHANG ; Chang-Jun WANG ; Xiang-Rong CAO ; Xiu-Zhen PAN
Journal of Medical Postgraduates 2018;31(5):453-458
Objective Serine /threonine kinases (STK) and phosphatases (STP) regulate various physiological activities of prokaryotes by reversible phosphorylation of proteins .This paper aimed to study the effects of simultaneous deletion of the stk and stp1 genes on the biological characteristics and pathogenicity of streptococcus suis type 2, the Chinese virulent strain 05ZYH33. Methods The double mutant of the stk and stp1 genes of 05ZYH33 was constructed by homologous recombination .The biological characteristics of the wild strain 05ZYH33 and the mutant strain Δstk/stp1 were compared.The effects of the stk and stp1 deletion on bacterial virulence was analyzed using cell adhesion assay , anti-phagocytosis assay and the mouse model of infection . Results RT-PCR showed that the stk and stp1 genes were replaced by the spectinomycin resistance gene Spc r and the mutant strain was successfully constructed .Experi-ments of biological characterization revealed gradually increased value of 05ZYH33 and Δstk/stp1 at 2 hours after inoculation and a plateau period at 7 hours.The logarithmic phase of the mutant strain (A600≈0.4) was 1 hour later than that of the wild one , and the bacterial den-sity of the former was lower than that of the latter after the plateau pe -riod (0.8 vs 1.0).On the blood plates of 05ZYH33 and Δstk/stp1 were observed greyish, round, semitransparent, wet and smooth-sur-faced tiny bacterial colonies , around which there were hemolysis rings with no significant differences in colony morphology and hemolytic ac -tivity.In the experiment on pathogenicity , the mice of the 05ZYH33 group all died within 12 hours while 9 of the 30 mice in the Δstk/stp1 group died within 12 hours and all died within 24 hours. Conclusion The simultaneous deletion of the stk and stp1 genes may mainly affect the regulation of the proteins associated with bacte -rial proliferation and division.
6.Alteration of methylation status of fragile histidine triad gene promoter in patients with myelodysplastic syndrome.
Dong-ming YAO ; Jun QIAN ; Wen-rong XU ; Jiang LIN ; Yun-wei JIANG ; Xia FEI ; Lan-xiu HAN ; Yali WANG ; Jian-nong CEN ; Zi-xing CHEN
Chinese Journal of Medical Genetics 2008;25(1):36-39
OBJECTIVETo study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.
METHODSMethylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.
RESULTSHypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).
CONCLUSIONFHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.
Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics
7.Prevalence and molecular analysis of β-thalassemia in children of Han ethnicity in Chongqing city.
Xiu-yun YAO ; Yu-mei ZHANG ; Zhen-zi QIN ; Rong FAN ; Lin ZOU ; Shi-ping CHEN ; Lei ZHANG ; Qian XIE ; Xiao-yun ZHONG ; Ying XIAN ; Jie YU
Chinese Journal of Pediatrics 2013;51(7):518-522
OBJECTIVETo investigate the incidence and the gene mutation frequencies and patterns of β-thalassemia (β-Thal) in ethnic Han children in Chongqing city.
METHODA total of 1726 children were screened by using automatic hemocytic analyzer, cellulose acetate electrophoresis and fetal hemoglobin alkali denaturation test. Samples with mean corpuscular volume (MCV) < 80 fl, cell hemoglobin content (MCH) < 27 pg and hemoglobin A2 (HbA2) levels >3.3%, fetal hemoglobin (HbF) >2% for β-Thal screening indicators. The positive samples of screening indicators were detected and identified by PCR-reverse dot blot method for 18 common β-Thal mutations in Chinese populations, unknown mutations samples were subjected to DNA sequencing analysis of the β-globin gene.
RESULTTwenty-five cases of β-Thal carriers were observed from the 1726 samples, with 24 cases of β-Thal heterozygote and one case of double heterozygote. Therefore, the β-Thal carrier rate was 1.51%. After 1726 peripheral venous blood samples analyzed by hematological parameters, 164 positive cases of β-Thal screening indicators were found, with the positive rate being 9.50% (164/1726). A total of 6 different gene mutations were detected, the four most common mutations were as the following: CD41-42, IVS-II-654, CD17 and beta E. These four mutations as the major types in this area accounted for 88.00% of all the mutations. In addition, one rare mutation of 5 'UTR; + (43 -40) was found, and one case of the hemoglobin variant of Hb Zurich was reported in Chinese people for the first time.
CONCLUSIONChongqing is a high risk region of the β-Thal. Epidemiological Data from the research was useul for the genetic counseling and the prevention of β-Thal major.
Asian Continental Ancestry Group ; genetics ; Blood Chemical Analysis ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Counseling ; Hemoglobins ; analysis ; genetics ; Hemoglobins, Abnormal ; analysis ; genetics ; Heterozygote ; Humans ; Infant ; Male ; Mutation ; genetics ; Prevalence ; beta-Globins ; genetics ; beta-Thalassemia ; epidemiology ; ethnology ; genetics
8. Ethical issues in the development of medical artificial intelligence and solutions
Xiong HOU ; Yu-xiu LIU ; Lei QIAN ; Le XU ; Zhao-rong SHI
Journal of Medical Postgraduates 2019;32(10):1080-1083
The introduction of artificial intelligence into the medical field is also facing social ethical challenges while promoting changes in the field of health services. To realize the healthy development of medical artificial intelligence, we must consider the ethical issues inherent in the application of artificial intelligence technology in medical services. The article analyzes the ethical issues brought about by medical artificial intelligence from the aspects of security, doctor’s principal status, doctor-patient relationship, and fair benefit. By implementing special supervision, clarifying the principal status of doctors, and strengthening ethical protocols, the application of artificial intelligence in medical services will be guided to promote artificial intelligence to better improve the service capabilities and quality of healthcare system.
9.A Preliminary Study on the Construction and Visualization of Knowledge Graph for the Ancient Chinese Medical Book Ling Shu
Ying-Xuan CHEN ; Wei-Hao XIE ; Fan CHEN ; Qian XU ; Rong-Yao LI ; Zhen-Hu CHEN ; Xiu-Feng LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):782-790
Objective To explore the construction and visualization for knowledge graph of Ling Shu(Spiritual Pivot),with a view to providing ideas for the structured storage and display of the theoretical knowledge of the ancient Chinese medical books.Methods Using the professional idea of constructing knowledge graphs for reference,text mining technology was applied to construct the thesaurus,and then word division,entity recognition,and relationship extraction for the original text of Ling Shu were performed to get the elements of knowledge graph construction.The graph database Neo4j was used for the storage and query of the knowledge graph,and then the visual display of the knowledge graph was achieved.Results The 1 216 high-quality words consisting of the thesaurus of Ling Shu were obtained,and the construction of the knowledge graph of the theory of Ling Shu was realized.The constructed knowledge graph basically displayed the traditional Chinese medicine theories such as the correlation of visceral manifestations with essence qi,and the relationship between emotions and the five-zang organs described in Ling Shu,which made the retrieval and utilization of the related entities and relationships possible,and provided ideas for the structured storage and display of the theoretical knowledge of the ancient books of Chinese medicine.Conclusion The knowledge graph construction technology can be used to obtain the Chinese medicine theoretical knowledge graph of Ling Shu,and to display the knowledge connections of yin-yang and the five elements,and the internal organs and meridians expressed in the Ling Shu.The construction of the knowledge graph and its storage in the graph database enable the knowledge graph involved in the text of Ling Shu to be displayed in the form of visualized semantic network graph,and also make the embedding of other search systems such as the semantic search and semantic wiki possible,which will be helpful for the development of Chinese medicine intelligent medical services.
10.Prognosis and related factors of acute lymphoblastic leukemia.
Wei ZHANG ; Rong FU ; Wen-Hui LIU ; Yu-Qian CHENG ; Wen-Xiu SONG ; Li-Juan DU ; Er-Bao RUAN ; Li-Tong ZHANG ; Xiao-Ming WANG ; Yong LIANG ; Guo-Jin WANG ; Wen QU ; Jia SONG ; Rong-Li ZHANG ; Jing GUAN ; Li-Juan LI ; Peng ZOU ; Zong-Hong SHAO
Journal of Experimental Hematology 2007;15(5):1102-1106
In order to analyze the prognosis and related factors of acute lymphoblastic leukemia (ALL), 53 newly diagnosed ALL patients were enrolled in this study. The therapeutic efficacy and prognosis of 53 cases of ALL were analyzed, the remission, relapse, overall survival and event-free survival were studied, and relation between different factors and prognosis of ALL were investigated by comparison of cases in same stage. The results showed that the complete remission was achieved in 36 out of 53 patients, the total remission rate was 67.9%, the total relapse rate was 37.7%, the median relapse duration was 6 months after remission. Median overall survival (OS) and median event-free survival (EFS) time were 4 and 1 months after remission respectively, OS and EFS rate of 18 month was 35.1% and 14.2%. The patients with different gender had significantly different EFS. Age was an independent risk factor of CR rate. White blood cell count and hemoglobin level of newly diagnosed patients were significantly correlated with OS and EFS. Absolute neutrophil count (ANC) at the end of the induction chemotherapy was an independent related factor of OS, the higher ANC, the lower risk of death. The patients with or without chemotherapy related infection had different relapse rate. The patients with bleeding after chemotherapy had lower OS when compared with those without bleeding. Serum glucose level was a significant negative prognostic factor. It is concluded that there is higher relapse rate, poor prognosis in adult ALL in comparison with children. In order to decrease the relapse rate and prolong the EFS, individual therapeutical regimens and prophylaxis of complicating diseases should be applied to ALL patients.
Adolescent
;
Adult
;
Aged
;
Antineoplastic Combined Chemotherapy Protocols
;
therapeutic use
;
Child
;
Female
;
Humans
;
Male
;
Middle Aged
;
Neoplasm Recurrence, Local
;
prevention & control
;
Precursor Cell Lymphoblastic Leukemia-Lymphoma
;
diagnosis
;
therapy
;
Prognosis
;
Young Adult