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 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 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 identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.

Objective To observe the therapeutic effect of electronic moxibustion apparatus and stone needle for primary dysmenorrhea disease in 35 cases.Methods 70 patients were randomly divided into the treatment group of 35 cases by random number table ,using electronic moxibustion apparatus and stone needle for treatment;and con-trol group of 35 cases,only using electronic moxibustion apparatus for treatment from 5 days before menstruation ,the treatment was continuous for 5 days,a course including 3 menstruation cycles .The clinical therapeutic effect after one treatment course was observed .Results The curative effects of the two groups were respectively 91.42% and 88.57%(χ2 =0.16,P>0.05);recovery rates of the two groups were respectively 77.14%and 42.85%(χ2 =8.57, P<0.05).The scores of the VAS,CMSS of the treatment group were much lower than those of the control group after treatment (t=2.532,2.137,2.256,P<0.05).Conclusion Electronic moxibustion apparatus and stone needle therapy is an effective method to treat primary dysmenorrhea ,which is a safe and convenient method .

Objective To retrospectively review the PET/CT imaging features of sarcoidosis and improve the diagnostic accuracy of this benign disease.Methods The PET/CT imaging characteristics and clinical data, including lesion size, distribution, standardized uptake value (SUV) and the ratio of misdiagnosis, of 11 sarcoidosis patients (5 confirmed pathologically and 6 clinically) were retrospectively analyzed.Results (1) Eleven patients had lymph node involvement:mediastinum and hilar lymphadenopathy in 11/11, supraclavicular fossa lymphadenopathy in 8/11, retroperitoneal lymphadenopathy in 8/11, pelvic cavity lymphadenopathy in 3/11.(2) Extrathoracic lesions were found in 7/11 with 4 lung involvement, 2 liver involvement, 1 parotid gland and temporalis involvement and 1 bilateral iliac and sacral bone involvement.(3) The size of the lesions ranged from 1.0 to 4.6 cm and the CT density ranged from 30 to 40 HU.The lesions in the lung are hypodense and in the liver are slightly hypo-or iso-dense.18F-fluorodeoxyglucose (FDG) uptake of all lesions was definitely increased in 6 cases; 18F-FDG uptake of some lesions was moderately or definitely increased in 2 cases, and slightly increased uptake in 3 cases.(4) The PET/CT diagnosis was consistent with the final diagnosis in 6/11.The 5 cases of misdiagnosis were malignant lymphoma (4/11) and lung cancer ( 1/11 ).Conclusions Differentiation between sarcoidosis and lymphoma in patients presenting with hilar lynphadenopathy can be difficult.Whole-body PET/CT may be helpful in the differentiation of the two diseases.