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 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 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.

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.

Objective To improve the diagnosis and management of solid-pseudopapillary tumor (SPT) of the pancreas. MethodsTwenty-two SPT patients were retrospectively reviewed and divided into two groups, misdiagnosed group and those with preoperatively correct diagnosis. ResultsAbout one half (46%) SPT cases were misdiagnosed preoperatively. With time SPT tends to invade its capsule resulting in impossibility of radical resection, and increased medical expenses. ConclusionsAlthough SPT is of low degree malignant, and the prognosis after surgical resection is satisfactory, misdiagnosis and preoperative misdiagnosis and inappropriate management still cost the patients increased expenses and inhospital stay.

AIM:To investigate the effect of Polo-like kinase-1(Plk1) depletion on cell cycle progression and cell growth in lung cancer cells.METHODS:A recombinant plasmid containing antisense RNA targeting Plk1(pcDNA3-Plk1) was transfected into A549 cells by lipofectine.RT-PCR and Western blotting were used to examine Plk1 gene expression.Cell proliferation was evaluated by cell counting and BrdU labeling.Cell cycle distribution and apoptosis were examined by flow cytometry.Inhibition rate(IR) of vinorebline(NVB) was determined by MTT assay.RESULTS:After transfected with pcDNA3-Plk1 into A549 cells,the expression levels of Plk1 mRNA and protein were greatly decreased.Abnormal morphological changes of cells and growth inhibition were observed in pcDNA3-Plk1 transfected cells.The BrdU labeling index was significantly lower than that in control group(P

AIM: To detect the changes of serum vascular endothelial growth factor(VEGF) level and the expression of peripheral blood cytokeratin 19(CK19) mRNA in patients with non-small cell lung cancer(NSCLC).METHODS: 96 patients with NSCLC,40 patients with benign lung diseases and 25 healthy controls were investigated.The VEGF level in serum was detected by ELISA and CK19 mRNA in peripheral blood was determined by using reverse transcriptase-polymerase chain reaction(RT-PCR).RESULTS: In NSCLC group,the serum VEGF level and the positive rate of CK19 mRNA in peripheral blood were(346.3?95.6) ng/L and 63.5%,which were significantly higher than those in other two groups respectively(P0.05).VEGF serum level in the patients who showed positive CK19 mRNA in peripheral blood was(407.4?121.2) ng/L.It is significantly higher than that in the negative patients(P

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.