1.Correlations between the Expressions of SRSF1 and Survivn and the Pathological Features of Prostate Cancer
Bangming XIAO ; Xingwang ZHU ; Liming DONG ; Yili LIU ; Ping WANG
Journal of China Medical University 2016;45(5):452-455
Objective To test the expression of serine/arginine rich splicing factor 1(SRSF1)and apoptosis inhibiting factor(Survivin) in prostate cancer,and study their correlation with the pathological features of prostate cancer,so as to put forward the new targets in the treatment of prostate cancer. Methods SRSF1 and Survivin protein was determined in 20 prostate tissue samples including prostate cancer(n=12)and benign prostat?ic hyperplasia(n=8)by immunohistochemical SP method. SRSF1 and Survivin was correlated to pathological features,and both the relevance was analyzed(no related reports at home and abroad). Results The positive expression rate of SRSF1 protein in prostate cancer tissue cells was 76.37± 5.06%,which was significantly higher than that of benign prostatic hyperplasia(11.30%±1.09%,P<0.05);the positive expression rate of Survivin protein in prostate cancer tissue cells was 86.93%±3.21%,which was significantly higher than that of benign prostatic hyperplasia(17.67%±1.99%, P<0.05);SRSF1 and Survivin protein expressed in prostate cancer organizations and were positively correlated to pathological Gleason grading, and there was significant correlation(P<0.05). Conclusion SRSF1 and Survivin protein were highly expressed in adenocarcinoma tissue,which were significantly increased than that of benign hyperplasia of prostate tissue. The positive expression SRSF1 and Survivin protein were positively cor?related to pathological Gleason grading.The expression of Survivin protein was elevated with the expression of SRSF1 protein in prostate cancer. These preliminary evidence indicated that SRSF1 may up?regulate the expression of Survivin,and thus promote the occurrence and development of prostate cancer.
2.Analysis on the gene mutations of MYOC in primary open angle glaucoma pedigree
Xiaobing XIE ; Xin ZHOU ; Yanli TIAN ; Xiying QU ; Duoxiu KUANG ; Huibin ZHU ; Jingcheng YU ; Xingwang NING
Chinese Journal of Laboratory Medicine 2009;32(2):157-161
Objective To screen the mutations of MYOC gene in a Chinese primary open angle glaucoma (POAG) family from Cbengqing and investigate the relationship between the mutations in MYOC/TIGR gene and POAG.Methods In a large 4-generation glaucoma family, myocilin gene (MYOC) was screened in 39 family members, 8 of which were confirmed patients. Normal controls included 100 normal Chinese subjects.The known mutations of MYOC gene ( including G34C, C136T, G144T, G227A, C624G,G736A, C1009G, A1036G, C1081T, G1099A, G1138A, A1139C, T1430A, C1441A and C1442T) were detected by single strand conformation polymorphism(SSCP) , po]ymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and DNA sequencing.Results G227A mutation was detected in 2 POAG patients and 1 asymptomatic patient, but not in the controls.Cl009del mutation was identified in all patients of the pedigree and an offspring member but not in the controls. No other mutations were detected.Since the C1009del mutation was revealed for the first time, a new GenBank number FJ237047 correponding to ACI62293 was applied.Conclusions The G227A mutation is a known site and there is no relationship between G227A mutation and glaucoma. But C1009del may be related to glaucoma which suggests that morbidity could be higher in the relatives of POAG than the controls.
3.Comparison of two HBV DNA detection kits
Haiying ZHANG ; Ying JI ; Ling ZHU ; Huiying RAO ; Jianghua WANG ; Henghui ZHANG ; Xingwang XIE ; Lai WEI
Chinese Journal of Laboratory Medicine 2011;34(5):459-464
Objective To evaluate clinical significance of two real-time fluorescence quantitative PCR kits for quantitative detection of HBV DNA and detection performance at different viral load levels.Methods A series of calibrators with different concentrations(1×106,5×105,1×105,5×104,1×104,5×103,1×103,5×102,1×102,1×101 kIU/L) were prepared with AB-type sera using the second generation WHO international standard (NIBSC code:97/750). HBV viral load in the sera of 78 patients,30 healthy blood donors and 10 calibrators were detected by real-time fluorescence quantitative PCR HBV DNA test kit from PIJI Bio-Technical Development Company Ltd (PG kit) and Cobas AmpliPrep/Cobas TaqMan HBV test kit. The correlation of the two methods was evaluated, and the performance of the two kits different viral load levels was evaluated. The false negative rate was analyzed. Negative control, low positive control and high positive control were included in every batch. Results Both two kits showed the correct results for the 10 specimens from the WHO international standards. The lowest detection limit of HBV DNA for Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit were 2.00 (kIU/L, lg) and 3.00 (kIU/L,lg) ,respectively. There was linear correlation between the results from Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit ( R2=0.938 7, P < 0.01 ), the upper limit of Roche kit had coincided with theoretical value. The samples with HBV DNA level above the upper limit of detection were diluted and retested to obtain the precise result. The result form Roche Cobas AmpliPrep/Cabas TaqMan HBV test [(8.35±0.20) kIU/L, lg] was higher than that from PG kit [(7.73±0.42 ) kIU/L, lg] (t=3. 776, P <0.05) . The detection of 108 serum samples showed that the level of HBV DNA detected by Roche Cobas AmpliPrep/Cobas TaqMan HBV test [(5.88±1.64) kIU/L, lg] was higher than that by PG kit [(5.25±1.55 kIU/L,lg] (t=12. 297 ,P <0.01 ). The correlation coefficients were high in samples with high HBV viral load[HBV DNA(>5.00 and≤7.00) kIU/L,Ig,R2=0. 779 7, P <0.01 ;HBV DNA( >7.00 ands≤9.00) kIU/L,lg,R2=0.603 7, P <0.01]. The correlation coefficient was low in samples with low HBV viral load[HBV DNA ( > 3.00 and≤5.00) kIU/L, lg, R2=0. 417 3, P <0.01 )]. When HBV DNA ( >3.00 and≤4.00) kIU/L,lg,the false negative rate was 33.3% (5/15). When HBV DNA ( > 1.08and≤3.00) kIU/L,lg,none of positive samples was detected with PG kit. Conclusions PG kit is not as good as Cobas AmpliPrep/Cobas TaqMan HBV test . The linear correlation between the results from the two kits is good. The correlation between the results detected with PG kit and Cobas AmpliPrep/Cobas TaqMan HBV test is higher in the high viral load groups than in the low viral load group. It is suggested that PG kit had a narrower linear range.
4.Simultaneous Extraction and Determination of Eighteen Fluoroquinolone, Tetracycline and Sulfonamide Antibiotics from Soils Using Solid-phase Extraction and Liquid Chromatography Tandem Mass Spectrometry
Lili MA ; Changsheng GUO ; Wei HU ; Jian SHA ; Xingwang ZHU ; Yuefei RUAN ; Yuqiu WANG
Chinese Journal of Analytical Chemistry 2010;38(1):21-26
An analytical method was developed for the simultaneous extraction and determination of eighteen fluoroquinolones (FQs), tetracyclines (TCs) and sulfonamides (SAs) antibiotics from soils using solid phaseextraction and liquid chromatography tandem mass spectrometry.Soil sample was firstly extracted by phosphate buffer at pH 3 in combination with 50% of organic modifier acetonitrile, then purified and concentrated by SAX and HLB column.Qualitative and quantitative analysis were carried out for the analyte under the MRM mode after the chromatography separation on Kromasil C_(18)(250 mm x4.6 mm, 5 μm) column.The range of recoveries (in percent) for FQs, TCs, SAs, in the soil matrix was 67.20%-88.98%, 62.23%-85.36%, 55.76%-97.37% with 1.1%-17.2% of relative standard deviation respectively in two different concentra tions.The limits of quantification (LOQ, S/N = 3) were 3.36-8.88 jig/kg, 0.56-0.91 μg/kg and 0.07-1.85 μg/kg for FQs, TCs and SAs, respectively.This method was successfully used to detect 18 anti biotics in 6 soil samples with different land types in Tianjin.Results showed some of the antibiotics in the arable soil were detected, with concentrations of 1.72-119.57 μg/kg.
5.Clinical features and emergency management of severe encephalitis and neurogenic pulmonary edema caused by enterovirus type 71 in children
Yuncai ZHANG ; Xingwang LI ; Xiaodong ZHU ; Suyun QIAN ; Jiansheng ZENG ; Yunxiao SHANG ; Biru LIU ; Xiaolin LIU ; Xiangui RAN
Chinese Journal of Emergency Medicine 2008;17(12):1250-1254
Objective To investigate the clinical characteristics and emergency management of severe hand-foot-mouth disease(HFMD)associated with encephalitis and neurogenic pulmonary edema(NPE)caused by en-terovirus 71(EV71)in children.Method Data of critical patients with severe HFMD associated with encephalitis and NPE admitted to pediatric intensive care unit(PICU)Fuyan city Hospitals Anhni Province from May to June 2008 were reviewed.Results Of 30 patients,the mean age was 15.8 months ranged from 4 months to 48 months.The overall morality was 19.4%.Tha average duration of critical symptoms persisted Was 2.1 days ranged from 12 hours to 5 days.There were no rash found in 12 patients(33.3%).The chinical features of nervous system mani-fested the symptoms of brainstem encephalitis in 27 patients(75%),brainstem encephalitis with myelitis in 6 pa-tients(16.7%),and encephalitis in 3 patients(8.3%).The frothy expectoration tinged with pink or bloody,asyrmmetrical pulmonary edema or hemoptysis were the main features of NPE.The main approaches to the treatment were mechanical ventilation,mannitol,methylpredifiselone,intravenous immunoglobulin(IVIG),and vasoactive a-gents.And nine patients(25%)needed fluid volume resuscitation in addition.Conclusions Young children are particularly vulnerable to the Severe EV71 encephalitis with NPE.The majority of involved fatal patients are aged under 3 years.Patients may die of acute onset of NPE and/or hemoptysis with rapid progress towards cardiopul-monary failure.Early diagnosis and evaluation,respiratory support,lowering intracranial pressure and maintaining hemodynamics ale the essential therapeutic approaches.
6.Effect of Low Rabll Expression on the Proliferation and Invasion of Human Bladder Cancer Cells
Xue GONG ; Liu YU ; Xingwang ZHU ; Jia LIU ; Yili LIU ; Ping WANG
Journal of China Medical University 2018;47(3):247-250,259
Objective To investigate the effect of inhibition of Rabll expression on the proliferation and invasion of human bladder cancer cell line T24. Methods T24 cells were transfected with Rabll siRNA, and RNA interference efficiency was determined by performing Western blotting. The effect of inhibition of Rabll expression on cell proliferation, cell cycle progression, and cell invasion was analyzed by performing CCK8, cell cycle detection, and Matrigel invasion assays, respectively. Expression of cell cycle-related proteins cyclin D1 and cyclin E and invasion-related protein matrix metalloproteinase 9 (MMP9) was determined by performing Western blotting and RT-PCR. Results Inhibition of Rabl 1 expression inhibited the proliferation and invasion of bladder cancer cells and downregulated the expression of cell cycle-related proteins cyclin D1 and cyclin E and invasion-related protein MMP9. Conclusion Our results suggest that Rabl 1 functions as a tumor protein and is involved in the progression of bladder cancer.
7.MRI changes of component and morphology in the internal carotid vulnerable plaque
Bao CUI ; Xingwang SUN ; Yufen ZHU ; Yuping DU ; Jinguo CUI ; Jianming CAI
Chinese Journal of Radiology 2018;52(4):295-299
Objective To investigate the changes of component and morphology in internal carotid vulnerable plaque,for helping to make clinical intervention strategy individually. Methods A total of 47 patients with internal carotid vulnerable plaques and primary hypertension underwent 2 high-resolution and multi-contrast MRI scans, from March 2008 to April 2014 were retrospectively reviewed. At baseline, the plaque was mainly located at the proximal internal carotid artery,and maximum plaque thickness ≥1.5 mm with intraplaque hemorrhage(IPH)and(or)thin or ruptured fibrous cap.Interscan interval was 0.5 years and above. Patients with carotid occlusion or surgery were excluded. Morphological measurements included maximum plaque thickness, maximum plaque area and cross-sectional vessel area (CSVA) on the level of plaque with maximum thickness. The paired-samples t test was performed to compare the difference of plaque morphology between baseline and follow-up carotid MRI.Results The interscan interval was 1.83 (1.59,1.99)years for 47 internal carotid vulnerable plaques.One case(interscan interval 2.16 years)showed IPH within those 11 plaques without IPH at baseline,and one case(interscan interval 1.42 years)had new incident IPH within those 36 plaques with IPH at baseline. Maximum plaque thickness increased significantly from(3.94±1.44)mm to(4.24±1.68)mm(t=2.30,P<0.05)by 5.14%(-3.83,11.34)% per year. Maximum plaque area increased significantly from(49.19±21.15)mm2to(56.03±24.91)mm2(t=3.87,P<0.01)by 6.67%(-2.26,19.60)% per year.CSVA increased significantly from(66.22±27.51)mm2to(73.68±31.47)mm2(t=4.08,P<0.01)by 5.18%(-1.63,12.34)% per year.Conclusion The progression of component,burden and outer remodeling in the internal carotid vulnerable plaque may be faster in hypertension, therefore reasonable intervention strategy and regular follow-up carotid MRI should be performed.
8.The significance of PCT and SAA detection in early childhood bacterial infection in preschool children
Xingwang NING ; Min SU ; Chun QIAN ; Xiaobing XIE ; Huibin ZHU
International Journal of Laboratory Medicine 2018;39(8):925-928
Objective To investigate the clinical significance of procalcitonin(PCT)and serum amyloid A (SAA)in early bacterial infection in preschool children.Methods 67 children with bacterial infection(bacte-rial infection group),62 children with viral infection(viral infection group)and 60 healthy children(healthy control group)were enrolled in this study,latex enhanced turbidimetric immunoassay was used to detect SAA and turbidimetric immunoassay was used to detect the level of PCT,the levels of SAA and PCT and the posi-tive rate were compared among all groups,and the sensitivity,specificity and positive predictive value,negative predictive value and Youden index of SAA and PCT levels.Results The levels of SAA and PCT in the bacte-rial infection group before treatment were significantly higher than those in the viral infection group and the healthy control group(P<0.05),7 days after treatment,the levels of SAA and PCT decreased significantly (P<0.05);the level of PCT in viral infection group was not significantly different from that in healthy control group(P>0.05),but the positive rate of SAA was significantly different from that of the healthy control group(P<0.05).The sensitivity,specificity,positive predictive value and negative predictive value of PCT for early bacterial infection in preschool children were 92.5%,93.5%,93.9%,92.1%,respectively,of the SAA values were 97.0%,59.7%,72.2% and 94.9%,respectively,there were significant differences between the two groups in specificity and positive predictive value(P<0.05).Conclusion Detection of SAA and PCT lev-els was helpful for early diagnosis,differential diagnosis and prognosis of bacterial infection in preschool chil-dren.
9.Analysis of the molecular epidemiological characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Hunan Province
Xingwang NING ; Yongxue TANG ; Siyu WANG ; Xiaomei WANG ; Huibin ZHU ; Xiaobing XIE ; Qingyu LIU
Chinese Journal of Preventive Medicine 2024;58(7):1041-1047
To examine the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) and investigate the horizontal transmission of blaKPC and blaNDM genes for the prevention and treatment of CRKP. A total of 49 clinically isolated CRKP strains were retrospectively analyzed from January to December 2022 at The First Hospital of Hunan University of Chinese Medicine. Phenotypic screening was performed using modified carbapenem inactivation assay (mCIM) and EDTA-carbapenem inactivation assay (eCIM). Polymerase chain reaction (PCR) was utilized to identify carbapenem resistance genes, β-lactamase resistance genes, and virulence genes, while multi-locus sequence analysis (MLST) was employed to assess the homology of CRKP strains. Conjugation experiments were conducted to infer the horizontal transmission mechanism of blaKPC and blaNDM genes. The results showed that the study included 49 CRKP strains, with 44 carrying blaKPC and 8 carrying blaNDM, Three strains were identified as blaKPC+ blaNDM-CRKP. In this study, 28 out of 49 CRKP strains (57.2%) were found to carry virulence genes. Additionally, one CRKP strain tested positive in the string test and was found to carry both Aerobactin and rmpA virulence genes. MLST results revealed a total of 5 ST types, with ST11 being predominant (41/49, 83.7%). Successful conjugation was observed in all 3 blaKPC-CRKP strains, while only 1 out of 3 blaNDM-CRKP strains showed successful conjugation. The transconjugant exhibited significantly reduced susceptibility to imipenem and cephalosporin antibiotics. In conclusion, the resistance mechanism of CRKP in this study is primarily attributed to the production of KPC enzymes, along with the presence of multiple β-lactamase resistance genes. Additionally, there is a local prevalence of hv-CRKP and blaKPC+ blaNDM-CRKP. blaKPC and blaNDM can be horizontally transmitted through plasmids, with varying efficiency among different strains.
10.Analysis of the molecular epidemiological characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Hunan Province
Xingwang NING ; Yongxue TANG ; Siyu WANG ; Xiaomei WANG ; Huibin ZHU ; Xiaobing XIE ; Qingyu LIU
Chinese Journal of Preventive Medicine 2024;58(7):1041-1047
To examine the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) and investigate the horizontal transmission of blaKPC and blaNDM genes for the prevention and treatment of CRKP. A total of 49 clinically isolated CRKP strains were retrospectively analyzed from January to December 2022 at The First Hospital of Hunan University of Chinese Medicine. Phenotypic screening was performed using modified carbapenem inactivation assay (mCIM) and EDTA-carbapenem inactivation assay (eCIM). Polymerase chain reaction (PCR) was utilized to identify carbapenem resistance genes, β-lactamase resistance genes, and virulence genes, while multi-locus sequence analysis (MLST) was employed to assess the homology of CRKP strains. Conjugation experiments were conducted to infer the horizontal transmission mechanism of blaKPC and blaNDM genes. The results showed that the study included 49 CRKP strains, with 44 carrying blaKPC and 8 carrying blaNDM, Three strains were identified as blaKPC+ blaNDM-CRKP. In this study, 28 out of 49 CRKP strains (57.2%) were found to carry virulence genes. Additionally, one CRKP strain tested positive in the string test and was found to carry both Aerobactin and rmpA virulence genes. MLST results revealed a total of 5 ST types, with ST11 being predominant (41/49, 83.7%). Successful conjugation was observed in all 3 blaKPC-CRKP strains, while only 1 out of 3 blaNDM-CRKP strains showed successful conjugation. The transconjugant exhibited significantly reduced susceptibility to imipenem and cephalosporin antibiotics. In conclusion, the resistance mechanism of CRKP in this study is primarily attributed to the production of KPC enzymes, along with the presence of multiple β-lactamase resistance genes. Additionally, there is a local prevalence of hv-CRKP and blaKPC+ blaNDM-CRKP. blaKPC and blaNDM can be horizontally transmitted through plasmids, with varying efficiency among different strains.