Objective To investigate the influence of an anti-integrin α6 monoclonal antibody (GoH3) on the in vitro infection of a human keratinocyte cell line HaCaT with HPV6/11 virus particles (VP). Methods HaCaT cells were infected in vitro with 4 different concentrations of HPV6/11 VP alone, HPV6/11 VP of 106 copies/ml after incubation with 6 different dilutions of GoH3, or 8 clinical isolates of HPV6/11 VP of 106 copies/ml before or after the incubation with 1∶ 100 dilution of GoH3. After additional culture, the infected HaCaT cells were collected and fluorescence quantitative (FQ)-PCR was performed to detect the HPV DNA load in cells. The inhibition rate of CoH3 on the infection was calculated. Results The viral load was different among the HaCaT cells infected with different concentrations of HPV6/11 VP (P < 0.01). The inhibition rate on the infection positively correlated with the concentration of CoH3 when the dilution was more than 1∶ 100; however, when the dilution was less than 1∶ 100, the increase in CoH3 concentration had no influence on the inhibition rate. The average viral load in HaCaT cells infected with clinical isolates of HPV6/11 VP was (5.81 ± 2.51) × 104 copies/ml in the absence of GoH3, (3.02 ± 1.21) × 104 copies/ml with the presence of CoH3, and the average inhibition rate of GoH3 was (46.9 ± 4.7)%. Conclusions GoH3 could partially suppress the adhesion of HPV6/11 VP to HaCaT cells, hinting that integrin a6 is an important HPV6/11 VP receptor on host cells.

Objective To investigate human plasma glycoproteomie changes related to human immunodeficiency virus (HIV) infection,and to identify glycoproteins with potential anti-HIV activity or anti-HIV drug targets. Methods Plasma proteins with lower abundance were enriched through affinity purification to remove albumin and IgG in clinical samples (HIV-positive patient, n= 10, and healthy controls, n= 20). Proteins were separated by two-dimensional electrophoresis (2-DE) and stained by Pro-Q emerald glycoprotein stain kits. The 2-DE image was analyzed by ImageMaster software to find differential glycoproteins. Furthermore, the depleted HIV-positive and healthy control plasma proteins were digested by PNGase F. Glycoproteins were deglycoliszed, separated by 2-DE and analyzed by ImageMaster software. Differential glycoproteins were identified by liquid chromatography combined with high capacity ion trap mass spectrometry (HCT). Results The pretreatment of HIV-positive plasma prior to 2-DE could efficiently remove the high aboundant albumin and IgG in plasma and improve the detection of proteins with low-abundance. High revolution 2-DE gel images of glycoproteins from HIV positive and healthy control plasma samples were obtained. Glycoproteins were successfully deglycolized through PNGase F treatment. Thirteen differential glycoproteins were identified by liquid chromatography combined with mass spectrometry. These proteins included alphalantitrypsin precursor and serine/threonine-protein kinase N1. Conclusions Potential HIV infection related proteins,such as alphal-antitrypsin precursor are successfully identified. Our study may offer some help to understand the molecular mechanism of HIV infection and select new drug targets for preventing HIV infection.

Objective To investigate the expression of integrin α6 mRNA in skin lesions of patients with condyloma acuminata. Methods Sixty outpatients with condyloma acuminate, including 30 primary cases and 30 recurrent cases were enrolled in the study;30 non-condyloma acuminate patients undergoing circumcision were used as normal controls. The expression of integrin α6 mRNA in condyloma acuminata lesions of primary and recurrent cases, as well as in skin samples of normal controls were determined by RTPCR. ANOVA was used to compare the differences among the groups, and Student-Newman-Keuls test(q test)was used for pairwise comparisons. Results Relative contents of integrin α6 mRNA(integrin α6/GAPDH)in normal controls, primary and recurrent condyloma acuminata lesions were 0.25±0.10, 0.79±0.16 and 1.07±0.29, respectively, and the difference was of statistical significance(F=127.687, P=0.001). Conclusion Integrin α6 may be associated with the pathogenesis and recurrence of condyloma acuminate, which may provide a new target for prevention and treatment of the disease.

A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of lopinavir and ritonavir in human plasma. Analytes were separated from plasma by a combination of alkalinized protein precipitation and liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a Agilent ZORBAX Eclipse XDB-C18 column with the mobile phase consisted of methanol-0.1% formic acid in water (80:20). A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the positive ion mode. Quantification was performed using multiple reaction monitoring (MRM) of the transitions m/z 629.6 --> 155.2, m/z 721.4 --> 268.2, and m/z 515.2 --> 276.2 for lopinavir, ritonavir and telmisartan (internal standard), respectively. The method showed a good linearity in a concentration range of 62.5 - 10000 ng mL(-1) for lopinavir, and 12.5 - 2000 ng mL(-1) for ritonavir. The lower limits of quantification were 15 pg mL(-1) and 8 pg mL(-1) for lopinavir and ritonavir, respectively. The intra- and inter-day precision was less than 15% and the absolute recovery was above 75%. This method was selective and rapid, sensitive for investigating blood drug concentrations in clinics.

Objective To study the effect of corrected TIMI frame count (CTFC) of infarction related artery on systolic function of infarct area of myocardium after primary percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI). Methods One hundred and six patients with AMI having undergone successful PCI in Cangzhou Central Hospital were selected, and they were divided into two groups (each, 53 cases). The standard of fast or slow flow was in accord to the CTFC of infarction related artery (IRA) measured soon after successful PCI. The patients with greater value of CTFC were enrolled in the slow flow group, while the patients with smaller such value were assigned in the fast flow group. At 6, 12, 24 and 48 hours after PCI, the venous plasma MB isoenzyme of creatine kinase (CK-MB) level was measured. And at 1 week, 1 month and 3 months after PCI, the left ventricular ejection fraction (LVEF) was measured by cardiac ultrasound, and the levels of radial strain (RS) and longitudinal strain (LS) of the infarct area were measured via speckle tracking imaging (STI). The differences in CTFC, CK-MB, RS and LS between the two groups were analyzed, and the correlations between the strains and CTFC, CK-MB were analyzed by Pearson linear correlation method. Results After successful PCI, the CK-MB of fast flow group was higher than that of the slow flow group at 6 hours. However, the CK-MB of slow flow group was higher than that of the fast flow group after 12 hours, appearing separate phenomenon, and the statistical significance occurred beginning from 24 hours after PCI (U/L, 24 hours:98.43±11.65 vs. 86.43±18.97, 48 hours:51.09±8.94 vs. 49.80±6.92, both P<0.05). CTFC in fast flow group was significantly lower than that of slow flow group (frame: 22.69±4.83 vs. 26.14±5.67, P < 0.01). After 3 months of follow-up, LVEF in fast flow group was higher than that of the slow flow group, but the difference had no significance (P > 0.05). RS and LS in fast flow group were higher than those in slow flow group, and the statistically significant difference appeared from 1 month after PCI (1 month RS:29.74±6.66 vs. 26.86±5.61, LS:-16.37±3.91 vs. -15.27±3.22, 3 months RS: 30.03±6.31 vs. 27.63±5.67, LS: -17.74±3.96 vs. -15.75±4.17, all P < 0.05). Pearson linear correlation showed:the strains (both RS and LS) and CK-MB had no significant relation (both P>0.05). Both RS and LS at 1 week, 1 month and 3 months were of significantly positive correlation with CTFC of each group (fast flow group:r value of CTFC and RS was respectively-0.526,-0.515,-0.532, r value of CTFC and LS was respectively-0.532,-0.541,-0.572;slow flow group:r value of CTFC and RS was respectively-0.691,-0.685,-0.702, r value of CTFC and LS was respectively-0.621,-0.584,-0.605, all P<0.01). Conclusion CTFC has some relationship with the recovery of the systolic function in area of infarct myocardium after PCI, and can be regarded as an important index to predict the long-term prognosis in patients with AMI.

ObjectiveTo find specific biomarkers related to HAART treatment in plasma samples of AIDS patients for clinical therapeautic efficacy evaluation and guidance for the prognosis of HIV treatment.MethodPlasma samples of AIDS patients were collected from Infectious Disease Department 1 of Shanghai Public Health Clinical Center in June of 2008 to February of 2009,including 11 successfully HAART treated cases (HIV load ＞ 50 copies/ml) and 11 unsuccessfully HAART treated cases (HIV load ＜50 copies/ml).Patients' age ranged from 22 to 63.Plasma samples were treated by Bio-rad AurumTM Serum Protein Mini Kit to remove high abundant proteins:albumin and immunoglobulin were removed.The treatedplasmaproteinswereseparatedbytwo-dimensionalelectrophoresisandanalyzedby electrophoretogram using Imagemaster software to find differentially-expressed proteins related to therapeutic efficacy.After digestion by trypsin,the differentially-expressed proteins were identified by online reversed-phasenano-flow liquid chromatography coupled with electrospray ionization ion trap mass spectrometry.ResultsLow abundant proteins were efficiently enriched after the AurumTM Serum Protein Mini Kit treatment.Six differentially-expressed proteins were detected while comparing successfully and unsuccessfully HAART treated group.These proteins were accurately identified by tandem Mass spectrometry (MS), including serum transferrin, serum β-fibrinogen, etc.ConclusionsOur proteomic research revealed that the differentially-expressed proteins such as transferrin,which is related to plasma virus loading in AIDS patients in the process of treatment,might be potential biomarkers evaluating HAART therapeutic efficacy.

Objective To observe the therapeutic effect of Qili Qiangxin capsule combined with glucocorticoid for treatment of patients with dilated cardiomyopathy accompanied by refractory heart failure. Methods A prospective study was conducted. Forty-eight patients with dilated cardiomyopathy and refractory heart failure in Hebei Cangzhou Central Hospital were enrolled,and they were randomly divided into three groups:control group,treatmentⅠand treatmentⅡgroups(each,16 cases). All groups were treated with conventional anti-heart failure western treatment, meanwhile additionally prednisone was given to treatment groupⅠand groupⅡ,firstly 40 mg/d,then the dosage of 5 mg was decreased in every 5 days until reaching 5 mg per day;in treatment groupⅡ,besides the same treatment of group I,the traditional Chinese medicine therapy Qili Qiangxin capsule 4 granules(one capsule 0.3 g)each time and 3 times a day was added,2 months being the therapeutic course in all the patients. The clinical efficacy and cardiac functional indexes,such as the left ventricular end-diastolic volume(LVEDV),the left ventricular end-systolic volume(LVESV),the left ventricular ejection fraction(LVEF)and the plasma B type brain natriuretic peptide (BNP),etc. were observed in 1 week and 2 months after treatment. Meanwhile the electrocardiogram(ECG),aspartate transaminase(AST),alanine aminotransferase(ALT),urea nitrogen(BUN),serum creatinine(SCr),blood routine, urine routine examination and the adverse effects were investigated. Results The total effective rates in treatment groupⅠand treatment groupⅡwere significantly higher than those in the control group〔after treatment for 1 week:81.2%(13/16),81.2%(13/16)vs. 43.8%(7/16);after 2 months:87.5%(14/16),93.7%(15/16)vs. 50.0%(8/16), all P<0.05〕. After treatment,the LVEDV,LVESV and BNP were lowered and the LVEF was increased in the three groups,and the above indexes in treatment groupⅡwere improved more significantly than those in groupⅠ〔LVEDV (mL):142.4±33.0 vs. 174.8±52.5,LVESV(mL):111.6±23.7 vs. 132.4±29.0,LVEF:0.421±0.037 vs. 0.390±0.045,BNP(μg/L):1.944±0.751 vs. 3.038±1.905,all P<0.05〕. Conclusion Qili Qiangxin capsule combined with glucocorticoid may effectively improve the cardiac function and clinical symptoms in near and forward future in patients with dilated cardiomyopathy accompanied by refractory heart failure,thus it may elevate the patients' life quality.

To design teaching course of problem-based learning (PBL) combined with case-based learning (CBL) for standardized training of clinicians based on clinical decision thinking mode (CDTM) of diagnosis and treatment process for primary liver cancer. The CDTM of treatment choice for primary liver cancer is multi-scheme selection model. The general decision rule includes decision condition, action plan and decision tree. For the theoretical study of clinical decision rules, it is suitable to design PBL teaching, but it is suitable to design CBL teaching course for clinical decision-making practice. The teaching course of PBL combined with CBL is conducive to improving the ability of clinicians' clinical decision thinking step by step.

Objective To assess polymerase chain reaction(PCR)combined with restriction fragment length polymorphism(RFLP)and gene sequencing technologies in the detection and typing of HPV DNA.Methods Tissue specimens were collected from skin diseases and venereal disease in perianal or genitals.PCR was performed with HPV DNA general primers(MY09/11)in tissue samples. Positive fragments of HPV DNA were purified and digested by restriction enzymes.The digested fragments were typed by po]yacrylamide gel electrophoresis(PAGE).The Resultswere verified by direct sequencing.Results In 50 clinical samples there were 35 HPV DNA positive,including 26 from patients with condyloma acuminatum,8 from patients with bowenoid papulosis,and 1 from patients with squamous cell carcinoma.In HPV DNA positive samples,19 were HPV6,3 were HPV11,8 were HPV16,4 were HPV6 and HPV 11,and I was HPV62.Sequencing Resultswere in accordance with the PCR-RFLP Results .Conclusion PCRRFLP method is effective in the detection and typing of HPV DNA.

Objective To observe the effects of recombinant human B-type natriuretic peptide (rhBNP) on cardiac function and heart rate variability (HRV) in patients with heart failure after acute myocardial infarction (AMI). Methods One hundred and twenty patients with heart failure after AMI admitted to the Department of Cardiology of Cangzhou Central Hospital of Hebei Province from January 2015 to January 2018 were enrolled. The patients were divided into a conventional treatment group and an rhBNP treatment group according to random number table method, with 60 cases in each group. The two groups were treated according to the AMI guidelines, the conventional treatment group received west medicine anti-myocardial ischemia and anti-heart failure treatment; the rhBNP treatment group received rhBNP on the basis of routine treatment; the first load dose was 2 μg/kg intravenous injection impact treatment, followed by maintaining dose 8.5 ng·kg-1·min-1 intravenous drip for 7 days. The changes of hypersensitivity C-reactive protein (hs-CRP), N-terminal B-type natriuretic peptide precursor (NT-proBNP), left ventricular ejection fraction (LVEF) and HRV index were observed before and after treatment in the two groups [HRV indexes including the changes of average normal RR interval standard deviation (SDNN), the average value of the normal RR interval standard deviation (SDANN), the root mean square (RMSSD) of the adjacent RR interval difference, and the percentage of adjacent RR interval difference > 50 ms (PNN50)]; the incidences of adverse reactions in the two groups were observed. Results After treatment, the levels of hs-CRP and NT-proBNP in the two groups were significantly lower than those before treatment (all P < 0.05). LVEF, SDNN, SDANN, RMSSD and PNN50 were higher than those before treatment, and the changes of the above indicators in the rhBNP treatment group were more significant than those in the conventional treatment group [hs-CRP (mg/L): 6.2±3.3 vs. 11.8±5.5, NT-proBNP (ng/L): 2.5±2.0 vs. 6.4±4.3, LVEF: 0.49±0.02 vs. 0.44±0.04, SDNN (ms): 93.3±18.1 vs. 79.1±16.0, SDANN (ms): 87.3±17.8 vs. 70.9±14.9, RMSSD: 30.3±11.0 vs. 23.8±10.4, PNN50: (15.9±7.3)% vs. (9.6±5.5)%, all P < 0.05]; No significant adverse reactions occurred during the treatment of the two groups. Conclusion rhBNP can significantly improve the heart function of patients with heart failure after AMI, reduce the levels of inflammatory response indicators and improve HRV;since its clinical efficacy is good, and its application safe, it is worthy to promote its clinical use.