1.Present status and prospect of irinotecan application in adjuvant therapy for colorectal cancer.
Chinese Journal of Oncology 2006;28(7):553-554
Antineoplastic Combined Chemotherapy Protocols
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therapeutic use
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Camptothecin
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administration & dosage
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analogs & derivatives
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Chemotherapy, Adjuvant
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Colorectal Neoplasms
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drug therapy
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pathology
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surgery
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Disease-Free Survival
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Fluorouracil
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administration & dosage
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Humans
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Leucovorin
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administration & dosage
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Postoperative Care
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Treatment Outcome
2.Analysis of suspicious results of serum HBV DNA detected by fluorescence quantitative PCR
Zhanguo CHEN ; Wu ZHOU ; Zhongyong WANG ; Yalei JIN ; Zhihua TAO
Chinese Journal of Laboratory Medicine 2013;(3):217-221
Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71%) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65%) and 381 (19.35%),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87%,20.23%,51.33% and 59.57% respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13%,79.77%,48.67% and 40.43% respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49%,4.08%,10.09% and 14.47% respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.
3.Reverse hybridization applied in detection on human papillomavirus infection of twenty-three subfamilies
Jin-Cai HE ; Xiao-Mei ZHOU ; Tao HUANG ; Wei REN ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To establish a method of reverse hybridization to detect five subfamilies of low risk Human Papillomaviruses(HPV6,11,42,43 and 44)and eighteen subfamilies of high risk HPV (HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83 and MM4)in one reaction.Methods Special probes for twenty-three HPV subfamilies were fixed on nylon membrane bars,biotin labeled general primers mediated polymerase chain reaction(GP-PCR)were applied in HPV DNA amplification.PCR amplified DNA fragments were reversely hybridized with special probes that were fixed on the membranes. All samples(136)detected by reverse hybridization method were paralleled with the methods of Hybridization Capture Ⅱ(HC-Ⅱ)and sequencing.Results Positive rate of the 136 samples detected by reverse hybridization was 41.9%,while HC-Ⅱ 42.6% and sequencing 40.4%.Reverse hybridization detection indicated coherence with the other two methods(Kappa 0.8644 and 0.9089,respectively).While sequencing was lab standard for DNA test,the sensitivity was 96.36%,specificity was 95.06%,accuracy was 95.59%.Conclusions Method of reverse hybridization is adaptable to 23 kinds of HPV subfamilies, which can confirm the exactly subfamilies of HPV infection.This method is adaptable in clinical detection of HPV,with high sensitivity,high specificity,simply and convenient operation and the results are easily to be read.
4.Follow-up research and dosage correlation analysis in patient with clopidogrel hypo-responsiveness after percutaneous coronary intervention
Min LU ; Tao FAN ; Jianlong ZHOU ; Xiaoqi JIN ; Xiaodong SHENG
Chinese Journal of Interventional Cardiology 2016;24(4):216-220
Objective To depermine if a double mainpenance dose of clopidogrel can improve phe clinical oupcome in papienps who have clopidogrel htpo-responsiveness ( CH) afper percupaneous coronart inpervenpion (PCI) and analtze correlapive risk facpors of CH. Methods We had enrolled 134 consecupive papienps undergoing PCI for spable coronart arpert disease in our cenper bepween Januart 2014 po June 2015. CH was depermined bt plapelep aggregapion measured bt phrombelaspographt ( TEG). Blood samples were paken 24 h and 3 monphs afper PCI procedure. All subjecps were divided inpo 2 groups (i. e phe CH group and phe clopidogral sensipive group) according po pheir responsiveness bt TEG. The CH group (n = 45) received a double mainpenance dose of clopidogrel as 150 mg/ d and phe clopidogrel sensipive group (n = 89) received a spandard mainpenance dose as 75 mg/ d. Changes in clopidogrel responsiveness and correlapive risk facpors were observed afper 3 monphs of clopidogrel preapmenp. Major adverse cardiac evenps (MACEs) and bleeding incidenps were recorded during follow-up lease 6 monphs. Results The clopidoprel htpo-responsive rape decreased from 33. 6% (45 / 134 papienps) po 11. 9% (16 / 134 papienps) afper 3 monphs of preapmenp. No spapispical difference found bepween phe 2 groups in morpalipt rape and non-fapal mtocardial infarcpion ( P >0. 05). Rapes of overall MACE (33. 3% vs. 22. 5% ), rehospipalizapion (26. 7% vs. 16. 9% ) and pargep vessel revascularizapion (11. 1% vs. 6. 7% ) were significanp higher in phe CH group ( all P < 0. 05) . Mulpivariape regression analtsis showed: smoking ( OR 4. 498, 95% CI 1. 378 - 4. 018, P = 0. 036), diabepes (OR 4. 385, 95% CI 1. 370 - 7. 552,P = 0. 026) and clopidogrel dosage ( OR 0. 597, 95% CI 1. 005 - 2. 676, P = 0. 019 ) were phe risk facpors for CH. Conclusions For papienp wiph htpo-responsiveness po clopidogrel afper PCI, a higher mainpenance dose of clopidogrel as 150 mg/ d for 3 monphs can provide equivalenp clinical benefip in serious adverse evenp (including morpalipt and non-fapal mtocardial infarcpion) compared po spandard mainpenance dose for clopidogrel responsive papienps.
5.Effect of OAZ signaling blocking through small interfering RNA on the production of anti-nuclear antibody in systemic lupus erythematosus
Rongliang LI ; Jin HUANG ; Tao ZHOU ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2010;14(1):3-7
Objective To explore the role of OAZ gene in the pathogenesis of systemic lupus erythe-matosus (SLE) by using RNA interfering technique. Methods Peripheral blood mononuclear cells (PBMC) from SLE patients were collected. Each sample was equally divided into four groups for cell culture in 96 well plates. Specific siRNA for OAZ and GAPDH were concordantly added to the experimental group and the positive control group, while nonspecific siRNA was added to the negative control group and only culture medium was added to the Mock control group. Cells and supernatants were harvested after culturing for 72 hours, then RNA was extracted and reverse transcripted to cDNA. OAZ, Id1, Id2, Id3, Id4 mRNA expression levels were analyzed by using real-time PCR. Levels of IFN-γ, IL-4, IL-10, IL-12, IL-21, CCL2, ANA in the supernatant were tested by ELISA. Relationships between the expression levels of OAZ mRNA with levels of cytokines and ANA were analyzed. Results OAZ, Id1, Id2, Id3 gene mRNA expression levels (△Ct: 12.5±1.4, 8.9±1.5, 4.3±0.8, 8.04±1.1) in the experimental group were significantly decreased comparing to those in the negative control group (△Ct: 10.2±1.1, 6.5±1.2, 2.4±1.3, 6.2±1.2 respectively, P<0.05). Levels of IFN-γ, IL-10, IL-12, IL-21 and ANA in the experimental group were significantly lower than those in the negative control group (P< 0.05), but level of CCL2 was higher than the negative control group (P<0.05). Difference of OAZ mRNA expression levels (△△Ct) between the experimental group and the negative control group was negatively correlated with changes of ANA, IL-21 levels, but positively correlated with changes of Th1/Th2, CCL2. Conclusion OAZ siRNA can effectively reduce the expression of genes involved in the OAZ signaling pathway in SLE. OAZ may lead to abnormal production of ANA via regulating Id genes and cytokines.
6.Expression and prognostic value of brain and acute leukemia,cytoplasmic in meningiomas
He YONG-TAO ; Zhou QIAO ; Zhu TAO ; Zhu JIA ; Zhang JING ; Jin MEI
Chinese Medical Journal 2019;132(18):2248-2250
7.Detection of carboxypeptidase H specific T cells in peripheral blood of latent autoimmune diabetic patients with carboxypeptidase antibody positivity by ELISPOT assay
Lin YANG ; Zhiguang ZHOU ; Tao DU ; Shaozhen TAN ; Yi ZHNAG ; Ping JIN
Journal of Central South University(Medical Sciences) 2009;34(10):1011-1016
Objective To explore the characteristics of T cell immunity in peripheral blood of patients with carboxypeptidase - H antibody (CPH-Ab). Methods Forty-two latent autoimmune diabetes in adults (LADA) patients with CPH-Ab~+ alone, 20 Type 2 diabetes mellitus patients (T2DM), and 22 healthy controls were selected and their peripheral blood mononuclear cells were isolated. Human recombinant carboxypeptidase (CPH) protein was expressed and further used as a stimulant in Enzyme-linked immunospot (ELISPOT) assay to detect IFN-γ-Th1 and IL-4-Th2 cells in the 3 groups. Th1/Th2 ratios were also calculated. CPH-Ab and glutamic acid decarboxylase antibody (GAD-Ab) were determined by radioligand assay. Results Compared with healthy controls and T2DM, IFN-γ-Th1 and IL-4-Th2 numbers did not increase significantly in CPH-Ab~+ group, nor did the Th1/Th2 ratios (P >0. 05). We further divided the CPH-Ab~+ patients into a short duration group (n = 22) and a long duration subgroup (n = 20) according to the duration of 3 years. CPH-IL-4-T in the short duration subgroup was significantly higher than that in T2DM and healthy controls (1. 8 vs. 0.2 and 0.3, both P < 0. 05) and we did not find any factor that was significantly correlated with the IL-4 spots number. There were not any significant differences in T cell responses to phaseolus vulgaris agglutinin (PHA) among all groups (P>0.05). Conclusion CPH does not directly involve in the cellular pathological mechanism of LADA. Anti-CPH immunity may be associated with more slowly aggressive beta cell autoimmunity.
8.Immune active effect of chemokine RANTES on human peripheral mononuclear cells
Xiao GU ; Hong ZHAO ; Jin YANG ; Guangchen ZHOU ; Shenyang GU ; Tao GU
Chinese Journal of Tissue Engineering Research 2008;12(40):7959-7961
BACKGROUND: Immune function of chemotaxis signal has been a key focus in medical research. However, the immune activation and related action mechanism of chemotatic factor RANTES are unclear.OBJECTIVE: To investigate the immune activation and related action mechanism of chemotatic factor RANTES stimulation on peripheral mononuclear cells.DESIGN: Control Experiment.SETTING: Department of Urologic Surgery, Clinical Medical College, Yangzhou University.MATERIALS: The experiment was performed at the Laboratory of Department of Immunology, University of Louisville from December 2004 to August 2005. Main reagent and equipments included RPMI 1640 complete medium, recombinant human RANTES, anti-CD3 monoclonal antibody, pyrrolidine dithiocarbamate(PDTC),CTLA4Ig, LS500 liquid scintillation counter and FACS Epics XL flow cytometry.METHODS: Peripheral mononuclear cells were collected and stimulated by different concentrations of recombinant human RANTES and/or anti-CD3 monoclonal antibody. Cells with significantly proliferative response were intervened by pyrrolidine dithiocarbamate (PDTC) or CTLA4lg. 3H-thymidine incorporation was used to detect the proliferation of mononuclear cells. Flow cytometry was applied to measure the phenotypes of lymphocytes.MAIN OUTCOME MEASURES: Incorporation efficiency of 3H-thymidine, the ratio of CD4 to CD8, expression of CD25,CCR5 and CD28.RESULTS: Proliferative reaction of mononuclear cells reached two peaks with recombinant human RANTES concentrations of 100 μg/L and 5000 μg/L respectively. The proliferation of peripheral mononuclear cells stimulated by 100 μg/L recombinant human RNATES was significantly higher than that in the presence of 50 μg/L anti-CD3 monoclonal antibody (P<0.05).There were no rivalry or synergistic effect between them.The immune active effects of recombinant human RANTES could be inhibited by PDTC or CTLA,Ig in a dose dependent manner.After RANTES treatment,the level of cell surface CD25 increased (P<0.05) and the CCR5 expression decreased (P<0.05), but there were no significant differences in CD28 expression and the ratio of CD4/CD8 of lymphocytes(P>0.05).CONCLUSION: RANTES has a specific function of inducing the immune activation of mononuclear cells. This special signal works depending on the activation of interleukin-2 signal pathway, CD28 co-stimulatory pathway and nuclear factor-κB,but independent of CD3 activation.
9.Effect of RNA interference on Polo- like kinase- 1 in A549 cells
Qiong ZHOU ; Yang JIN ; Xiaoju ZHANG ; Yuan SU ; Xiaonan TAO ; Ming BAL
Chinese Journal of Pathophysiology 2007;23(11):2185-2190
AIM: To investigate whether RNA interference (RNAi) induced by small interference RNA (siRNA) could suppress Polo- like kinase- 1 (Plk 1 ) expression and its effects in A549 cells. METHODS: A recombinant plasmid containing siRNA targeting Plk1 ( psiRNA - hH1 - Plk1 ) was transfected into A549 cells with Lipofectamine 2000.Expressions of Plk1, cyclin B1 and p53 protein were detected by Western blotting. Cell proliferation was evaluated by direct cell counting, while cell cycle and apoptosis were examined by flow cytometry, and expression of α - tubulin was detected by immunofluorescence. RESULTS: The results demonstrated that sequence specific siRNA targeting Plk1 was capable of suppressing Plk1 expression, and reflecting in lower kinase activity in A549 cells. The level of Plk1 protein was reduced by at least 70% after 48 h of psiRNA - hH1 - Plk1 treatment relative to controls. Expressions of cyclin B1 and p53 were increased greatly after Plk1 depletion, and cells showed absence of microtubule polymerization and spindle abnormalities in staining for α -tubulin. Growth inhibition, G2/M arrest and apoptosis were observed in psiRNA -hH1 -Plk1 transfected group. CONCLUSION: All these data suggest that siRNA targeted against human Plk1 may be a valuable tool in cancer therapy.
10.Changes of molecular markers of prothrombotic c state in plasma and puerarin for treatment of acute pancreatitis
Wei JIN ; Jian WANG ; Tao DONG ; Hongmei ZHOU ; Yongzhu LI ; Cunxin ZHAO ; Yinhua ZHANG
Clinical Medicine of China 2012;28(11):1160-1163
ObjectiveTo study changes of molecular markers of prothrombotic state:Platelet granule membrane protein ( GMP-140 ),Von Willebrand factor ( vWF:Ag),thrombomodulin (TM),Two-D dimer ( DD),antithrombin Ⅲ ( AT- Ⅲ ) in plasma and puerarin for treatment functions of acute pancreatitis (AP).MethodsIn 78 patients with AP [ severe acute pancreatitis (SAP):26 cases,mild acute pancreatitis (MAP):52 cases ],using a random number table,the patients were given puerarin treated base (n =40) and conventional treated base group (n =38 ).The two groups were given fast,continuous gastrointestinal decompression,correction of electrolyte and acid-base balance disorders,vein support,antisecretory drugs,antibiotics inhibit pancreatic secretion and inhibition of trypsin activity of drug treatment.Puerarin group:Puerarin injection 0.5 g in 5%glucose injection intravenous infusion of 500 ml,1 time a day.GMP-140 vWF:Ag,TM,DD were measured by the methods of analysis of enzyme-linked immunosorbent assay and AT-Ⅲ was measured by the methods of analysis of chromogenic substrate method preformed in all patients,plasma amylase and uric amylase were determined by the method of somogyi and after the treatment.And 22 healthy people were selected as normal controls ( NC,Group C,n =22).ResultsCompared with the Group C and MAP,the plasma GMP-140 [ ( 86.26 ± 15.28 )ng/Lvs (32.56 ± 18.17) ng/L and (58.68 ± 15.86)ng/L],vWF[(236.22 ±31.78)%vs (95.12 ±31.68)% and (126.68 ± 17.06)% ],TM [(65.70 ± 12.27) μg/L vs (4.26 ±0.92) μg/L and (9.80 ± 6.98) μg,/L],DD [ (0.87 ±0.04) mg/L vs (0.36 ±0.06) mg/L and (0.56 ±0.05) mg/L] were significantly elevated,however the AT-Ⅲ [ (56.13 ± 15.78) U/ml vs (98.76 ±22.68) U/ml and (80.38 ± 18.29)U/ml )was significantly decreased SAP ( P < 0.01 ).There were significant differences on the levels of GMP-140 [ (31.52 ± 15.81 ) ng/L vs (59.62 ± 13.73 ) ng/L,t =- 23.283 ],vWF [ ( 93.32 ± 28.62) % vs ( 128.81 ±16.23)%,t=-28.205,P<0.01 ],TM[ (4.36 ± 0.82) μg,/L vs (11.23 ± 7.62)μg/L,t =-43.419,P <0.001],DD[ (0.32 ±0.05) mg/L vs (0.68 ±0.04) mg/L,t =- 15.642,P <0.001],AT-Ⅲ ((97.68 ±21.69) U/ml vs (76.86 ± 17.92) U/m,t =14.967,P < 0.01 ) between puerarin treated base group and conventional treated base group.Comparing with treated base,the group given puerarin obviously shortened the increased of plasma [ ( 81.26 ± 17.12) U/L vs ( 119.63 ± 51.87 ) U/L,t =- 7.618,P < 0.001 ],uric amylase [ (416.37 ± 116.50) U/L vs (576.32 ± 126.58) U/L,t =- 36.659,P < 0.001 ],the time of abdominal pain relief and therapy to spend [ ( 2.18 ± 0.76 ) d vs ( 5.26 ± 0.58 ) d,t =- 13.619,P < 0.001 ].Conclusion The molecular markers of prothrombotic state:GMP-140,vWF:Ag,TM,DD,AT- Ⅲ might all play key roles in the development of AP.Puerarin can improve the pancreatic microcirculation and adjust molecular markers of prothrombotic state,and had certain treatment functions with AP.