Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Chemotherapy, Adjuvant ; Colorectal Neoplasms ; drug therapy ; pathology ; surgery ; Disease-Free Survival ; Fluorouracil ; administration & dosage ; Humans ; Leucovorin ; administration & dosage ; Postoperative Care ; Treatment Outcome

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.

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.

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.

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.

[Objective]To investigate the genetic polymorphisms of the CYP2C19 gene in the elderly Chinese Han populations of Guangzhou,and compare the frequencies of CYP2C19 gene polymorphisms in different populations,in order to provide accurate data for the appropriate prescription.[Methods]To detect the genetic polymorphisms of the CYP2C19 gene by the DNA microarray,and compare the frequencies of CYP2C19 gene polymorphisms in Chinese Han populations from different areas and the different races.[Results]There were 2312 case samples in our study. The allele frequencies of CYP2C19*1,CYP2C19*2 and CYP2C19*3 were 64.27%,30.75%,and 4.98%,respectively. As the genotype,EM(*1/*1)was 41.44%(n=958),IM(*1/*2,*1/*3)was 45.67%(n=1056),and PM(*2/*2,*2/*3 and*3/*3)was 12.89%(n=298). The ratios of EM and IM in Chinese Han populations from different areas and all the subtypes of the CYP2C19 genotype in different minority were statistically significant. As the races,there were difference in all the subtypes of the CYP2C19 genotype when Asian populations were compared with white races(P<1304.64)and black races(P<0.01),which was also statistically significant.[Conclusions]The distributions of the CYP2C19 gene polymorphisms were significantly different in Chinese han populations and in different races,and the main subtypes of the CYP2C19 genotype in the elderly of Chinese han populations were IM and EM,which is beneficial for prescribing appropriate in the elderly populations.

Objective To study the expression of plasma membrane Ca2 + -ATPase isoforms 1 -4 and the splice variants at sites A and C in the neonatal rat vestibular organ.Methods Ten rats at postnatal 2 days (P2 ) were decapitated and their vestibular organs (macula utriculi and macula sacculi)were isolated.The total proteins of the vestibular organs were extracted.The expression of PMCA1-4 splice variants at sites A and C was detected by RT-PCR.Results The splice variants of PMCA1-4 at sites A and C in macula utriculi and macula sacculi of neo-natal rat vestibular organs were PMCA1x/b,PMCA2w/(a,b),PMCA3z/(a,b,c)and PMCA4 (x,z)/b.Conclusion The splice variants at sites A and C among PMCA1,PMCA2,PMCA3 and PMCA4 were different in the vestibu-lar organs of neonatal rats,which could be explained that macula utriculi and macula sacculi had different require-ments of Ca2 + turning for these PMCA isoforms.

Purpose To analyze the gait of patients following reconstruction of posterior cruciate ligament(PCL) and posterolateral corner(PLC).Methods Between March 2007 and April 2008,16 patients with combined PCL and PLC injuries-deficient knee underwent the reconstruction with allograft.Dial test were performed before surgery,and gait analysis and dial test were assessed 1 year after reconstruction.Sixteen healthy volunteers were used as controls.Results There are no differences in the time-distance factors and kinematics between patients and controls(P>0.05).The torque of knee extension revealed significant difference between patients and controls(P<0.01).Mean abtorsion angle of the patients increased 16.5°±6.2° before surgery;whereas the angle increased-4.4°±7.8° 1 year after surgery(P<0.01).The maximal mean abtorsion angnlation from gait analysis was 14.1°±15.7° and from dial test was 29.7°±1 5.2°,respectively.There was a strong linear correlation(r=0.9671,P<0.05)between the outcomes from dial test and gait analysis.Conclusion After reconstruction of the PCL and PLC,the gait of patients became almost normal 1 year after operation,except the torque of knee extension.

Background and purpose:The risk of recurrence for colorectal cancer after curative surgery is up to 30%-40%. We aimed to evaluate the relationship between time to relapse (TTR) of colorectal cancer with clinical pathological parameters and overall survival after recurrence. Methods:We carried out the analysis of clinical data, pathological examination and follow up information of 375 colorectal cancer patients who admitted to Liaoning Cancer Hospital. Patients were categorized into relapse at<2, 2-5 and>5 years following their initial surgery. Results:TTR was associated with the clinical stage at diagnosis and liver or lung metastasis status. Short TTR (<2 years) was positively associated with survival. However, there was no significant difference in survival between patients who relapsed at 5 years or later compared with those who relapsed between 2 and 5 years. Conclusion:TTR within 2 years is an important predictor of shorter survival for colorectal cancer patients who experienced a relapse.

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.