Objective To establish a method to detect viral integrity of human papillomavirus in women cervical HPV infection. Methods We amplified E6/E7 gene and E2 gene of HPV16,then inserted them into a plasmid containing single copy HBB gene. HPV16 infected cervical epithelium samples were screened out by genotyping with RDB of flow-through hybridization assay.Fluo-rescence quantitive PCR data of HBB,viral E2 gene and viral E6 gene of all samples were standardized by compared with respective parameters of the plasmid.The ratio IHPV and CHPV were calculated to find out E2 gene disruption and viral copies per cell in the cer-vical samples,respectively.Results The plasmid constructed for standardization was proved effective to make the FQ-PCR data of E2 gene,E6 gene and HBB gene comparable.Thirty-seven HPV16 positive cervical epithelium samples included 22 cases from women whose TCT were normal,and 15 cases from women who confirmed HIL/CIN 2-3 or above through colposcopic examina-tion plus biopsy.Fifteen samples were detected E2 gene disruption,including 10 HIL/CIN 2-3 or above samples and 5 TCT normal samples.E2 gene integrity in different groups were statistically significant different(P <0.05).The average viral copies per cell dis-played a significant decline along with E2 gene disruption(P <0.05).Conclusion The tandem single copy gene plasmid standard-ized methord for the detection of E2 gene disruption caused by viral integration in HPV16 infected cervical cells is feasible and effec-tive.

Objective To establish a Goto-Kakizaki ( GK) rat model of duodenal-jejunal bypass( DJB) and ob-serve the changes in insulin-resistance after surgery, and to explore the mechanism of DJB surgery in treatment of type 2 di-abetes mellitus.Methods Male Goto-kakizaki diabetic rats(GK,n=36)were used as experiment group and 18 healthy male Wistar rats as blank group.GK rats were randomly divided into two groups: diabetic control group and DJB surgery group ( n=18) .Euglycemic-hyperinsulinemic clamp technique was performed in 6 rats randomly taken from each group at third week, sixth week and ninth week after surgery, respectively.The expression levels of Gck, G6P, PEPCK mRNA in the liver and GLUT4 content on skeletal muscle cell plasma membrance were detected one week after the clamp test. Results In the DJB surgery group at the end of third week and sixth week after surgery, the levels of glucose infusion rates and the expression levels of Gck, G6P, PEPCK mRNA in the liver showed no statistically significant difference as com-pared with the diabetic control group (P>0.05).In the DJB surgery group at the end of 9th week after surgery, the glu-cose infusion rate and expression level of Gck mRNA in the liver were significantly higher, and the expression of G6P and PEPCK mRNA was significantly lower than those in the diabetic control group ( P<0.05 for all) .At the three time points (3rd week, 6th week and 9th week after surgery), there was no significant difference in the GLUT4 content on skeletal muscle cell plasma membrane between the diabetic control group and DJB surgery group ( P>0.05 for all) .Conclusions Our results indicate that the mechanism of DJB surgery improving blood glucose level may be closely related to the amel-ioration of insulin-resistance in the liver, thereby augmenting glucose uptake and inhibiting gluconeogenesis in liver through the regulation of glucose metabolism-related enzymes.There is no significant improvement in insulin-resistance in the skele-tal muscles during the experiment period.This result implies that the effect of DJB surgery on type 2 diabetes is related to the duration of therapy.

OBJECTIVE:To study in vitro anticancer activity of conjugated linoleic acid-gemcitabine conjugate (CLA-GEM). METHODS:IC50 of different tumor cells (breast cancer MCF-7 cell,breast cancer MDA-MB-231 cell,lung cancer A549 cell, small cell lung cancer NCI-H446 cell,glioma C6 cell)were investigated after treated with different concentrations(0.001-100μmol/L)of CLA-GEM and gemcitabine(GEM)for 72 h;survival rates of MCF-7 cell were investigated after treated with above so-lution for 24,48 and 72 h. The dependence of 0.001-100 μmol/L CLA-GEM and GEM to nucleoside transporter was investigated (by IC50)through MCF-7 cells and MDA-MB-231 cells treated with nucleoside transporter inhibitors(NBMPR,100 μmol/L)and di-pyridamole(4 μg/ml). The change of MCF-7 cell cycle was investigated after treated with 1 μmol/L CLA-GEM and GEM for 24 h. RESULTS:Compared with GEM,IC50 of MCF-7,MDA-MB-231 and NCI-H446 cells became lower after treated with CLA- GEM (P<0.01),there were no statistical significances in IC50 between A549 and C6 cells(P>0.05). Survival rate of MCF-7 cells de-creased significantly after treated with GEM for 48 h and CLA-GEM for 24 h. Survival rate of MCF-7 cells was the lowest,being 21% after treated with GEM for 72 h,while tumor cells were sacrificed by CLA-GEM completely. Compared with GEM or CLA-GEM,IC50 of MCF-7 and MDA-MB-231 cells increased significantly after treated with NBMPR,dipyridamole combined with GEM (P<0.01);there were no statistical significance in IC50 after treated with NBMPR,dipyridamole combined with CLA-GEM (P>0.05). Compared with GEM,CLA-GEM could prolong 6% of S stage of MCF-7 cells (P<0.01). CONCLU-SIONS:Compared with GEM,CLA-GEM exhibits significant antitumor activity and rapid action,and it isn’t influenced by nucle-ic acid transportation.

Objective To investigate the distribution of human papillomavirus (HPV) type 16 in women cervical infection in eastern Guangzhou, polymorphism of E6/E7 gene and association of gene dosage with disease progression. Methods Flow-through hybridization and gene chips were applied in HPV sub-type identification to screen out HPV-16 positive samples from cervical epithelium samples. HPV-16 E6/E7 gene was amplified through PCR with specific primers. The PCR products were cloned into pMD18-T vectors and fragments were determined through sequencing. Polymorphism analysis were performed through align-ment tools. Fluorescence quantitive PCR were used for the detection of viral E6 gene and L1 gene. Results Thirty-six (4.5%) HPV-16 positive samples were screened out through flow-through hybridization from 806 cervical epithelium samples. HSIL and above happened in 18 (50.0%) of the 36 HPV-16 positive patients. Within E6/E7 gene sequences from 7 selected samples, we found 15 sites with variances and 8 of them would cause coding amino acid change. HIL group (A, 11 cases) and LSIL group (B, 14 cases) possess significantly different gene dosage of both viral E6 gene and LI gene (P <0.05). The ratios of L1/E6 be-tween the 2 groups was not significantly different(P=0.19). Conclusion HPV-16 cervical infection oc-curs in 4.5% women (17-62 years old) in eastern Guangzhou. HIL or above accompany with half of the HPV 16 infected women. Viral load is probably associated with cervical HSIL, though L1/E6 ratios do not suggest viral integration.

Objective:To observe the expression of nuclear factor-kappa B( NF-κB) in rats with acute lung injury ( ALI) caused by oleic acid ( OA) and investigate the protective effect of puerarin ( Pue) in ALI. Methods:Twenty-four rats were randomly divided into three groups:the control group, OA group and Pue group. Morphological changes in lung tissues were observed under a light mi-croscope and the expression of NF-κB in lung tissues were examined. Results:Morphological changes in lung tissues showed that pul-monary inflammation was relieved in Pue group compared with that in OA group. Compared with the control group, the expression of NF-κB was increased obviously in OA group (0. 39 ± 0. 07 vs 0. 12 ± 0. 04, P<0. 05), and that was decreased obviously in Pue group (0. 24 ± 0. 05), which was still higher than that in the control group (P<0. 05). Conclusion:Puerarin can alleviate the inflammation reaction in ALI through inhibiting the expression of NF-κB.

Objective:To study the diagnosis value of Transcranial Doppler ultrasound for intracranial artery stenosis in patients with cerebral infarction.Methods:A total of 140 patients with acute cerebral infarction,transient ischemic attack and posterior circulation ischemic attack in Department of Neurology,Xinjiang cardiovascular and cerebrovascular disease hospital from October 2014 to October 2016 were selected as research object,CT angiography (CTA) and TCD detection were performed in all patients.Used CTA examination results as the gold standard,the detection results of intracranial artery stenosis in two groups were compared,the diagnostic value of TCD and the diagnostic results of TCD to the degree of bilateral middle cerebral artery (MCA) stenosis were analyzed.Results:CTA diagnosis showed that 140 patients had a total of 105 patients with intracranial artery stenosis,in the anterior and posterior circulation vessel of 1155 intracranial segments,CTA detection showed that 249 vessels were narrow,TCD detection showed that 236 vessels were narrow.Com-pared with CTA,TCD was better in the diagnosis of patients (Kappa value＞0.75).The diagnostic sensitivity and positive predictive value of TCD for MCA were the highest,which were 91.26％ and 93.07％,the consistency was the best (Kappa value =0.917).CTA detection showed that 210 MCA vessels had 103 stenoses,mild stenosis 17,moderate stenosis 41,severe stenosis 45,TCD detection showed that the stenosis was 101,mild stenosis 16,moderate stenosis 40,severe stenosis 45.The Kappa test showed that the diagnostic results of TCD to the degree ofMCA stenosis was better consistency compared with CTA (Kappa value=0.884.Conclusion:TCD has a high diagnostic value for cerebral artery stenosis in patients with cerebral infarction,and it is consistent with the diagnosis of CTA.

AIM: To develop and perfect the similarity algorithm on chromatographic fingerprints of traditional Chinese medicine(TCM). METHODS: The experimental and simulated data were used to investigate and verify the characteristics on the new and former methods on similarity evaluation for chromatographic fingerprints method. RESULTS: There are many strong points for new similarity algorithm in practice. CONCLUSION: The new method provides more rationality and practicability in TCM quality control and analysis compare to existing methods.

Objective To discuss current detection of food intolerance in health check-up populations and the influence of food intolerance on immune system and inflammation parameters.Methods Serum levels of 14 kinds of food-specific IgG antibodies were analyzed by using enzyme linked immunosorbent assay (ELISA) among 257 healthy check-up adults.White blood cell count,eosinophils,globulin,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),rheumatoid factor (RF),antinuclear antibodies (ANA) and T cell function were compared between the positive group and the negative group.Student's t test and Chi-square test were used for data analysis.Results Food-specific IgG antibodies were found in 134 participants (52.14％).The most common intolerant foods were egg,crab and milk.Moderate to severe food intolerance was caused by egg or milk.There was significant difference in eosinophils and serum globulin between the positive and the negative group (t =-0.07,x2 =8.91,both P ＜0.05).However,no difference was observed in white blood cell count,ESR,CRP,RF,ANA and T cell function between two groups (all P ＞ 0.05).Conclusion Eosinophils and serum globulin may significantly increased in people with food intolerance,although white blood cell count,ESR,CRP,RF,ANA and T cell function could be normal.

Objective To observe the protective effect of gradual ischemic postconditioning (IP) capable of improving reperfusion on reperfusion injury in patients with ST-segment elevation acute myocardial infarction (STEAMl) undergoing primary percutaneous coronary intervention (PPCI).Methods 102 in-patients with STEAMI undergoing PPCI in the Department of Cardiology in the First People's Hospital of Kunshan City Affiliated to Jiangsu University from February 2011 to August 2014 were enrolled in this study. They were divided into three groups by a random number table: IP group (32 cases), gradual IP group (30 cases) and routine reperfusion group (40 cases). In IP group, after the opening of the infarction related blood vessel, ischemic postconditioning within the first minute of arterial reperfusion was made through three episodes of 1 minute inflation and 1 minute pressure withdrawn of an angioplasty balloon, and then persistent reperfusion was carried out. In the gradual IP group, the patients received three times of gradual angioplasty balloon inflation and denation, 1 minute/1minute, 30 seconds/30 seconds and 15 seconds/15 seconds respectively, presenting the gradual change of IP time. In the routine reperfusion group, after the opening of blocked blood vessel, the patients underwent routine PCI to persistently recover the coronary artery blood supply. The changes of related lead ST segment regression (Sum-STR), incidence of reperfusion arrhythmia, corrected thrombolysis in myocardial infarction (TIMI) franle count (CTFC), peaks of MB isoenzyme of creatine kinase (CK-MB), left ventricular ejection fraction (LVEF) and frequency of adverse events in follow-up period were compared among the three groups.Results The baseline characteristics were comparable in three groups. The incidence of ventricular premature beats was significantly lower in gradual IP group than that in routine reperfusion group [30.0% (9/30) vs. 55.0% (22/40),P < 0.05], and although theincidence of ventricular premature beats was lower in IP group than that in routine reperfusion group [34.4% (11/32) vs. 55.0% (22/40)], no statistically significant difference was found (P > 0.05). The incidence of ventricular tachycardia was significantly lower in IP and gradual IP groups than that in routine reperfusion group [15.6% (5/32), 13.3% (4/30) vs. 40.0% (16/40), bothP < 0.05]. The incidences of ventricular fibrillation, bradyarrhythmia and sinus arrest were lower in IP group and gradual IP group than those in routine reperfusion group, but no statistically significant differences were found (allP > 0.05). In IP group and gradual IP group, the Sum-STR incidence, CTFC, CK-MB peaks were lower than those of routine reperfusion group [Sum-STR: (56.7±18.3)%, (57.3±21.5)% vs. (44.6±21.6)%; CTFC: 25.47±5.37, 24.46±6.41 vs. 31.62±7.56; CK-MB peaks (U/L): 126.3±78.5, 121.6±82.5 vs. 147.4±72.5; allP < 0.05], the effect of gradual IP group being the best among the three groups. The levels of LVEF were slightly higher in IP and gradual IP groups than the level in routine reperfusion group (0.507±0.042, 0.511±0.062 vs. 0.497±0.062), but no statistically significant difference was found (bothP > 0.05). In routine reperfusion group, one patient died because the ventricular fibrillation could not be corrected and another one died of no-reflow during operation. Each group had 1 patient died during the 4 weeks of follow-up after operation, in the routine reperfusion group, one died of refractory heart failure, and the cause of death of other two patients, one in IP group and another in gradual IP group, was considered due to subacute thrombosis in stent. Major bleeding events were not found in each group.Conclusion Gradual IP can ameliorate myocardial reperfusion injury more significantly in patients with STEAMI undergoing PPCI.

Objective According to the requirements of the College of American Pathologists (CAP),to validate the laboratory developed test-JAK2 V617F mutation.Methods This is a clinical laboratory diagnosis study.Fifty-seven peripheral blood and bone marrow samples were collected from the BCR/ABL-negative myeloproliferative neoplasms (MPN) patients in Huashan Hospital from Apr 2010 to Dec 2010.JAK2 V617F mutation was detected by fluorescence PCR combined with melting curve analysis.The accuracy,precision,analytical sensitivity,and anti-interference ability of the assay were validated.The Kappa test was used to evaluate accuracy.Coincidence rate was used to evaluate the repeatability.Paired t test was used to evaluate analytical sensitivity.Results There was a close agreement between the reference method(sequencing) and the test method (fluorescence melting curve analysis) (Kappa =0.89).The coincidence rate was 100％.The results of the assay was not affected by lipoprotein (＜ 24 mmol/L) or bilirubin (＜450 μmol/L).The detection limit of the assay was 5％.Conclusion JAK2 V617F mutation assay by fluorescence PCR combined with melting curve analysis could be applied to the clinical laboratory.