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Objective:To investigate the association between hepatic echinococcus granulosus infection and necrosis with gene polymorphism of interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α), and to identify the related factors at the gene level.Methods:A total of 106 patients with hepatic echinococcosis who underwent surgical treatment in the department of hepatobiliary surgery, People′s Hospital of Xinjiang Uygur Autonomous Region from January 2018 to December 2020 were selected. Patients with necrosis caused by hepatic echinococcus granulosus infection were selected as the observation group, and patients without necrosis caused by hepatic echinococcus granulosus infection were selected as the control group, with 53 cases in each group. The serum levels of IL-10 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA). The polymorphisms of IL-10 (-592, -1082) and TNF-α (rsl800630) were detected by polymerase chain reaction (PCR). The levels of IL-10 and TNF-α and their gene polymorphisms were analyzed.Results:The levels of serum IL-10 and TNF-α in the observation group were significantly higher than those in the control group (all P<0.05); There was significant difference in genotype and allele frequency of IL-10 (-592, -1082) and TNF-α (rsl800630) (all P<0.05). The serum IL-10 level of CC genotype patients with IL-10 gene -592C/A locus in the observation group was higher than that of CA+ AA genotype patients, with statistically significant difference ( P<0.05). The serum IL-10 level in patients with TT genotype at -1082T>A of IL-10 gene in the observation group was higher than that in patients with TA+ AA genotype, with statistically significant difference ( P<0.05). The serum TNF-α level in patients with CC genotype at rsl800630C/A locus of TNF-α gene in the observation group was higher than that in patients with CA+ AA genotype, with statistically significant difference ( P<0.05). Conclusions:The changes of IL-10 (-592, -1082) and TNF-α (rsl800630) gene polymorphisms may be associated with hepatic echinococcus granulosus infection and necrosis.
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Objective To establish the re-examination criteria of the Sysmex XN-9000 automatic blood cell analyzer pipeline suitable for this laboratory for ensuring the accuracy of detection results .Methods A total of 2250 whole blood samples were se-lected from inpatients ,outpatients and subjects undergoing physical examination .The Sysmex XN-9000 blood cell analyzer was a-dopted to conduct the detection .With the manual microscopic examination as the golden standard ,26 items of re-examination criteria were performed the verification and evaluation .The positive predicting value ,negative predicting value ,false positive rate ,false neg-ative rate and re-examination rate were performed by the statistics .Results In compariing the instrument alarm information with the microscopic examination results ,the positive predicting value was 91 .59% ,the negative predicting value was 98 .64% ,the false positive rate was 2 .09% ,the false negative rate was 1 .02% and the re-examination rate was 24 .84% .Conclusion The formulated re-examination criteria of the Sysmex XN-9000 blood cell analyzer is in accord with the characteristics of our laboratory ,which in-creases the detection efficiency ,prevents the missing detection and false detection and has clinical guidance significance .
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Objective To investigate the present situation of nursing professional environments in our hospital and provide theoretical basis for mapping out relevant countermeasures. Method A questionnaire survey was conducted among 338 nurses in our hospital by using the general information questionnaire and practice environment scale (PES). Result The dimensions of PES by their average scores in descending sequence included leadership and ability of nurse administers, doctor-nurse cooperation, foundation for high quality care nurses′participation in hospital affairs, and sufficient manpower and resources. Conclusion In order to provide good and healthy work environment for nurses, nursing administrators should provide enough nursing manpower resoures and apply excitation mechanism in their hospital.
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Objective To evaluate intraoperative tracing of sentinel lymph node(SLN) by fluorescence staining combined with dye dyeing.Methods A total of 174 patients with breast cancer were randomly divided into three groups : the group A with 57 patients receiving methylene blue (MB), the group B with 58 patients receiving indocyanine green(ICG) as the lymphatic mapping tracers,the group C with 59 patients receiving MB and ICG.The sentinel and axillary lymph node of level Ⅱ, Ⅲ was excised, followed by conventional histopathology.Results There was no significant difference among the three groups in the term of visualized detection rate (x2=2.96, P =0.241).There was statistical significant difference among three groups in the term of detected lymph nodes(F=15.34, P＜0.05).Comparing with the three groups, the number of detected lymph nodes of A and B group had no significant differences(P=0.07) ,the number of detected lymph nodes of C was higher than that of A and B group, and the difference was significant(P＜0.05).There was statistical significant difference among three groups in the term of SLN positive rate (x2 =6.75, P =0.039), and there was no significant difference among A and B group(P=0.915) ,SLN positive rate of C group was higher than than A and B group, and the difference was statistical significant (P＜0.05).Conclusion Intraoperative tracing of SLN by fluorescence staining combined with dye dyeing has the skin and subcutaneous reveal advantage.The use of ICG fluorescence and MB increases lymph node detection rate.
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Objective To study antimicrobial resistance of clinically isolated Escherichia coli (E.coli),the preva-lence of integrons in E.coli,and relation of integron with antimicrobial resistance of E.coli.Methods E.coli isola-ted from three hospitals of Guangdong Province from 2010 to 2012 were collected,and antimicrobial susceptibility testing was performed by Kirby-Bauer method;integrons were detected by polymerase chain reaction (PCR),and gene cassette was analyzed by sequencing.Results A total of 156 E.coli isolates were collected,antimicrobial sus-ceptibility testing showed that resistance rate of E.coli to most penicillins,cephalosporins,fluoroquinolones,amin-oglycosides and sulfonamides were over 50%;the resistance rate to antimicrobials 0.05),but compared with sensitive E.coli (9.09%,2/22),the difference was statisti-cally different (P<0.01 ).There were nine types of integron-drug resistant gene cassettes in the variable regions, most of which contained aadA and dfrA.Conclusion Antimicrobial resistance of E.coli is serious;high incidence of class I integrons are widely found in E.coli,and is closely related with drug resistance of E.coli,class I integrons mainly mediated aminoglycosides,sulfonamides and beta-lactams resistance.
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The multiplex polymerase chain reaction (PCR) technique was applied to detect the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) specific target cDNA fragments in the present study. The target cDNA fragments of SARS-CoV were synthesized artificially according to the genome sequence of SARS-CoV in GenBank submitted by The Chinese University of Hong Kong, and were used as simulated positive samples. Five primers recommended by World Health Organization (WHO) were used to amplify the fragments by single PCR and multiplex PCR. Three target cDNA fragments (121, 182 and 302 bp), as well as the three different combinations of any two of these fragments, were amplified by single PCR. The combination of these three fragments was amplified by multiplex PCR. The results indicated that the multiplex PCR technique could be applied to detect the SARS-CoV specific target cDNA fragments successfully.
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Base Sequence , Cloning, Molecular , DNA Primers , DNA, Complementary , Genetics , Molecular Sequence Data , Polymerase Chain Reaction , Methods , Severe acute respiratory syndrome-related coronavirus , GeneticsABSTRACT
Objective:The aim of this study was to explore the epidemiological characteristics of large intestine carcinoma in North Zhejiang.Methods:A retrospective review of clinical records and endoscopic findings of 339 large intestine carcinoma patients (detected from 5320 endoscopic examinees) throught past 8 years was executed.Results:General detection rate was 6.37%.Annual detection rate fluctuated between 4.60% and 8.79%.The detection rate for male was slightly higher than that for female.The ratio of male to female was 1.15∶1.The 41～60 years interval was the susceptible age interval (48.37%);rectum and sigmoid colon were the most frequently sites involved (71.38%).Most (89.38%) of the carcinomas were adenomatous histopathologically.Conclusions:There were susceptible age interval and topological sites for large intestine carcinoma.Most of them were adenomatous.Annual detection rate in this region through the past 8 years showed neither rise nor decline year by year.
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AIM:To study the blocking effect of shRNA on the expression of PSMA gene in LNCaP cell line by using shRNA eukaryotic expression vector.METHODS:Three pairs of DNA templates coding shRNA,synthesized against PSMA and cloned into the vector pSilencer 2.1-U6-neo,which was named pSilencer 2.1-U6-neo-shRNA,were identified by restriction endonuclease digestion analysis and DNA sequencing.LNCaP cells were then transfected with these three pSilencer 2.1-U6-neo-shRNAs and the negative control pSilencer 2.1-U6-neo-NC.After G418 selection,the cells were selected and the interfering effect was detected by RT-PCR and Western blotting.The biological behaviours of the transfected LNCaP cells were also tested.RESULTS:Restriction endonuclease digestion analysis and DNA sequencing results all showed that the 3 target segments were cloned into pSilencer 2.1-U6-neo vector respectively.After transfected into LNCaP cells,the inhibitory ratio of PSMA mRNA was 33.15%,9.26% and 41.97% respectively,and that of PSMA protein was 26.26%,6.47%,40.69% respectively.The p-shRNA3 was chosen to test the cell growth and its invasive power in vitro.The results showed that after interfering,the invasiveness of LNCaP cells were enhanced.CONCLUSION:The vector-based shRNA on PSMA gene effectively knocks down the PSMA gene expression.The successful construction of PSMA shRNA makes it possible for further study of the interaction between PSMA and prostate cancer.
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AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.
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AIM: To discuss the relationship between prostate specific membrane antigen(PSMA) and prostate cancer and to seek a target for diagnosis and therapy of prostate cancer.METHODS: A pair of primers was designed according to the published PSMA mRNA sequence.Total RNA was extracted from prostate cancer tissues and was reversely transcribed into cDNA,which was used as a template for PCR to amplify the PSMA gene.The recombinant was sequenced and the result was analyzed by BLAST.The PSMA5 gene specific primers were designed to identify its expression in different cells and prostate tissues.RESULTS: A new alternatively spliced variant of PSMA named PSMA5 was discovered when sequencing the recombinant.PSMA5 showed well pathological tissue-specificity,and its expression rate in prostate cancer,benign prostatic hyperplasia of prostate,and normal prostate tissue were 92.6%,78.8% and 10.0%,respectively.It expressed specifically in Pca cell line LNCaP,not in cell lines of PC3,bladder carcinoma,renal carcinoma,or hepatoma.CONCLUSION: A new alternative spliced variant of PSMA named PSMA5 was discovered,which was well correlated with prostate cancer and benign prostatic hyperplasia.This finding may give a new clue to the evolution of prostate cancer and may provide a target for the diagnosis and therapy of prostate cancer.
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AIM: To obtain eukaryotic expression vector of Chinese prostate-specific membrane antigen. METHODS: Chinese prostate-specific membrane antigen (PSMA) cDNA was amplified by RT-PCR from prostate cancer tissues, then cloned into eukaryotic expression vector pcDNA3 0 and sequenced. RESULTS: Seven bases in Chinese PSMA cDNA sequence were found different from those reported by Israeli, which lead to two different amino acids. CONCLUSION: We have obtained the PSMA cDNA, and the recombinant eukaryotic expression vector was successfully constructed. The study lays foundation for DCs vaccine modified by PSMA gene for the treatment of prostate neoplasms.