Objective To analyze the genotypes and age distributions of human papillomavirus in affiliated hospital of Chifeng u‐niversity ,to provide theory basis for the screening and prevention of cervical cancer in Chifeng area .Methods The cervical exfolia‐ted cell specimens from 1 368 gynecological outpatients and female inpatients in affiliated hospital of Chifeng university were collect‐ed from January to June 2016 .Using PCR‐reverse dot blot technology ,25 HPV subtypes was performed ,a statistical analysis was conduted combing with age .Results Among 1 368 specimens ,546 specimens were found with positive HPV ,the positive rate was 39 .91% .The 24 genotypes were detected .The top three subtypes of HPV infection were HPV16(12 .08% ) ,HPV 58(8 .05% )and HPV 52(7 .61% ) ,HPV73 genotype was not detected .Masculine gender rate in the groups with various age had significance differ‐ence after the chi‐square test ,among different age groups ,the prevalence rate was the highest in the patients with higher than 56 years old(52 .5% ) ,followed by the rate in patients with lower than 25 years old(52 .38% ) .Conclusion The subtype‐specific and age‐specific distributions of HPV infection in Chifeng area have obvious heterogeneity ,which indicates that HPV genotypes and age should be considered in screening ,prevention and treatment of cervical cancer .

ObjectiveTo search the appropriate experimental conditions for using green fluores-cent protein (GFP) as a reporter gene in tumor gene therapy. Methods The plasmids carrying mu-tated GFP gene were transfected into the eukaryotic cells to observe transient gene expression. Thesestudies were conducted to 1. directly determine GFP expression and express stability in the COS- 7cells, 2. compare the transfection efficiency of two plasmids pcDNA3 - EGFP, pSVKa - S65T with dif-ferent promoters in different tumor cell lines, 3. determine two genes expression in a single cell usingLacZ cotransfected with GFP by FACS. ResultsFluorescence could be detected in intact viable cellsunder different sets of conditions. The expression of GFP might last two weeks or more and the ex-pressed fluorescence was stable. The transfection rate of pcDNA3 - EGFP expressed was different inthree tumor cell lines examined. But pSVK3 - GFP expressed similarly in four tumor cell lines exam-ined. FACS showed the probability of two genes entering a single cell is above >85% at the ratio 1:4.ConclusionThe above data indicate that the GFP can be visualized continuously and directly for geneexpression in living cells. GFP may also be used for quicklly selecting cells carrying a target gene.

Background Idiopathic macular epiretinal membrane (IMEM) shows a high incidence and affects visual function of patients.The primary management of IMEM is vitrectomy,but the study on prediction of visual acuity after operation by optical coherence tomography (OCT) is seldom.Objective This study was to evaluate the predicting ability of OCT for preoperative visual outcome for IMEM patients who received vitrectomy.Methods The clinical data of IMEM patients with vitrectomy from March 2009 to May 2013 were retrospectively analyzed in People's Hospital Peking University.Forty-nine eyes of 48 cases were divided into continuous group (19 eyes of 18 cases) and discontinuous group (31 eyes of 31 cases) based on whether the extraretinal layers were intact on OCT,including retinal pigment epithelium (RPE),inner and outer segment of photoreceptors (IS/OS) and external limiting membrane (ELM).Central foveal thickness (CFT) by high-resolution spectral-domain OCT and best corrected visual acuity (BCVA) (LogMAR) before surgery and 12 weeks after surgery were recorded and compared between the two groups,and the correlations of shift of CFT with the shift of LogMAR vision was analyzed by unitary linear regression to evaluate the value of OCT in predicting postoperative vision.Writen informed consent was obtained from each patient prior to any medical operation.Written informed consent was obtained from each patient before surgery.Results In 12 weeks after operation,reflective strips of RPE layer,IS/OS and ELM layer were intact in 18 eyes of the continuous group after vitrectomy.The number of eyes with discontinuous strip in RPE layer decreased to 1 eye after operation from 6 eyes before operation,and the number of eyes with IS/OS reflective strip discontinue decreased to 19 eyes after operation from 29 eyes before operation,and that with ELM layer discontinue decreased to 15 eyes after operation from before operation in the discontinuous group.The improvement of LogMAR BCVA was increased with the decline of CFT values with a negative correlation between them (R2 =0.298,B =0.001,P =0.000),and postoperative LogMAR visual acuity improved 1 line whenever CFT reduced by 100 μm.LogMAR BCVA was 0.4±0.19 before surgery and 0.36±0.21after surgery in the continuous group,showing a insignificant difference between them (t =0.876,P=0.393).In the discontinuous group,the postoperative IogMAR BCVA was significantly higher after operation (0.46±0.26) than 0.82 ±0.41 after operation (t =6.206,P =0.000).The increase value of LogMAR after vitrectomy was 0.3 in the discontinuous group,which was superior to 0.0 of the continuous group.Conclusions The continuity of extraretinal layers and CFT by OCT are closely associated with the improvement of vision after vitrectomy in the eyes with IMEM.Postoperative vision is better in IMEM eye with continuous retinal reflective strips before operation,and majority of IMEM eyes can improve vision after operation.Preoperative OCT seems to be an important diagnostic tool for the selection of patients who benefit or not from surgery to certain extent.

The recombinant plasmids pGenesil-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells and cisplatin was added with different concentrations in order to study the inhibitory effects of Livin gene, increase the apoptosis induced by cisplatin, and detect the expression of Bcl-2, Bax, caspase-3, and survivin genes. The pGenesil-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells, and the expression levels of Livin, Bcl-2, Bax, caspase-3, and survivin genes were detected by using fluorescence quantitative real-time PCR. Then cisplatin at different concentrations (3.0, 6.0 and 9.9 microg/mL) was added into the transfected Hela cells, and 24, and 48 h later, the apoptosis rate was measured by flow cytometry. After transfection of pGenesil-1-BIRC71 and pGenesil-1-BIRC72 into Hela cells, the expression level of Livin gene was obviously reduced, and the apoptosis rate was significantly increased in transfection group as compared with control group (P<0.05). Cisplatin could increase the apoptosis rate in a dose- and time-dependent manner. After cisplatin was added, the expression levels of Bcl-2 mRNA were reduced, and those of Bax, caspase-3, and survivin mRNA were increased in transfection group as compared with those in control group (P<0.05). It was concluded that shRNA expression vector targeting Livin gene could inhibit the expression of Livin gene in Hela cells and enhance the apoptosis induced by cisplatin, which was related to the decreased expression of Bcl-2 and activation of Bax and caspase-3. Survivin might play an important role as an antagonist in the process of apoptosis induction.

BACKGROUND:Basic fibroblast growth factor (bFGF) can enhance fibroblast proliferation and col agen deposition, and vascular endothelial growth factor (VEGF) can improve blood perfusion and metabolic level of pathological tissues. Additional y, both of them can boost the alkaline phosphatase activity under given conditions. OBJECTIVE:To evaluate the effect of bFGF combined with VEGF on the periodontal ligament fibroblast proliferation and alkaline phosphatase activity in rats. METHODS:Rat periodontal ligament fibroblasts were cultured in vitro, its embryonic origin was identified and passage 4 cel s were used for the fol owing experiments. Effects of bFGF and VEGF with different concentrations on the rat periodontal ligament fibroblast proliferation were detected to determine the minimum and maximum effective concentrations. Cel s were divided into five groups:group A (control group) with DMEM containing 2%fetal bovine serum;group B as maximum effective concentration of VEGF group;group C as maximum effective concentration of bFGF;group D as minimum effective concentration of bFGF combined with minimum effective concentration of VEGF group;group E as maximum effective concentration of bFGF combined with maximum effective concentration of VEGF group. At 3, 7 and 14 days, the alkaline phosphatase activity in each group was detected. RESULTS AND CONCLUSION:Rat periodontal ligament fibroblasts derived from the mesoderm grew wel . Rat periodontal ligament fibroblast proliferation was increased with the VEGF and bFGF concentration increasing (P<0.01). The maximum and minimum effective concentrations of VEGF were 100 and 10μg/L, and the maximum and minimum effective concentrations of bFGF were 10 and 0.1μg/L. The absorbance values in the groups D and E were higher than those in the group A. The absorbance values of the group D were significantly lower than those of the group E at 3 and 7 days (P<0.05), but did not significantly differ at 14 days (P>0.05). To conclude, the combination use of the maximum effective concentration of VEGF and bFGF can play a significant synergistic effect on the alkaline phosphatase activity at a given time, but the minimum and maximum effective concentrations show no significant differences if not in the given time, which may be related to the time-effectiveness of these two factors and the receptors of periodontal ligament cel s. Subject headings:Fibroblast Growth Factor 2;Vascular Endothelial Growth Factors;Alkaline Phosphatase;Tissue Engineering

Objective To investigate influences of paeonol on mRNA expression and function of drug transporters BCRP and SLCO1B1 in HepG2 cell.Methods Cell counting Kit-8 assay was used to detect the viability of HepG2 cells;Real-time fluorescent quantitative PCR (qPCR) was performed to measure the expressions of BCRP and SLCO1B1 mRNAs; flow cytometry was applied to determine the transport functions of BCRP and SLCO1B1. Results Paeonol (2-8μg/mL) did not decrease HepG2 cell survival rate, but 16 μg/mL paeonol significantly reduced cell survival rate (P<0.05). Paeonol(2-8μg/mL)significantly induced the mRNA expression and function of drug transporters BCRP and SLCO1B1(P <0.05).Compared with control group, transcription level of paeonol group’s BCRP and SLCO1B1 drug transporters obviously up-regulated, the of translocation efficiency of substrate specificity increased significantly (P <0.05).Conclusion Paeonol can induce drug hepatocellular transporters BCRP and SLCO1B1 gene expression, thereby promote the substrate transport the transmembrane.It is indicated that the drug combination of paeonol and BCRP and SLCO1B1 transporters, there may be a risk of drug interactions.

Objective To discuss the relationship between C-reactive protein and prognosis of acute coronary syndrome. Methods C-reactive protein of 60 acute coronary syndrome patients was evaluated. The individuals were divided into two groups:One group with higher CRP level and another group with normal CRP level. The following-up duration was 6 months. After correct therapy, the morbidity of re-angina, arrhythmia, heart failure, re-infarction and cardiac death was compared. Results The morbidity of re-angina, arrhythmia, heart failure, re-infarction, cardiac death was 46.3%(19/41), 43.9%(18/41),9.76%(4/41),22.0%(9/41),7.32%(3/41)respectively in higher CRP level group; The morbidity of re-angina, arrhythmia, heart failure, re-infarction was 15.8%(3/19),10.5%(2/19), 5.3%(1/19),5.3%(1/19)respectively in normal CRP group and there was no cardiac death accident. There was significant different between the two groups. Conclusions CRP plays an important role in the onset of acute coronary syndrome, and its level is related with the higher morbidity of re-angina, arrhythmia, heart failure, re-infarction and cardiac death.

Histopathological examination of testes is important in assessing spermatogenesis and testicular function. Modified Davidson's fluid (mDF) has been proposed as a superior substitute for Bouin's fluid (BF) for fixation of adult animal testes. Besides, 4% paraformaldehyde (PFA) has been commonly used to fix testes with convenience. We compared the morphology of the rat testis fixed in 4% PFA, mDF, or BF using hematoxylin and eosin (HE)-stained sections. Fixation in 4% PFA resulted in obvious tissue shrinkage artifacts, especially between seminiferous epithelium cells. Shrinkage artifacts were also observed in the central area of the testes fixed in BF. Use of mDF did not cause shrinkage artifacts between seminiferous tubules, though a small amount can be observed in seminiferous tubules between germ cells. Clarity of nuclear detail in testes fixed in mDF and BF is better compared to 4% PFA. Our study demonstrated that fixation in mDF provided better morphologic details in the rat testis as compared with 4% PFA and BF.

Objective To evaluate the role of spinal extracellular signal-regulated kinase (ERK) signaling pathway in reduction of remifentanil-induced hyperalgesia by electro-acupuncture (EA) at Zusanli in rats with incisional pain.Methods Fifty male adult Sprague-Dawley rats, weighing 250-280 g, in which the intrathecal catheter was successfully placed without complications, were randomly divided into 5 groups (n=10 each) using a random number table: control group (group C), remifentanil + incisional pain group (group RI), EA at acupoint group (group E) , EA at non-acupoint group (group NE), and 1/2 ERK inhibitor U0126 + EA at acupoint group (group UE).Normal saline 0.1 ml · kg-1 · min-1 was infused intravenously for 60 min in group C.In RI, E, NE and UE groups, after the model of incisional pain was established, remifentanil 1.0 μg · kg-1 · min-1 was infused for 60 min, and in addition, EA (intensity 10 mA, frequency 4 Hz) of Zusanli lasting for 60 min was performed at the same time in E and UE groups, and EA was performed at the points 5 mm lateral to the acupoints of Zusanli on the operated side simultaneously in group NE.ERK1/2 inhibitor U0126 5 μg (in 5％ dimethyl sulfoxide 10 μl) was injected intrathecally in group UE, and 5％ dimethyl sulfoxide 10 μl was injected intrathecally in the other groups.The mechanical paw withdrawal threshold (MWT) was measured before remifentanil or normal saline infusion (T1) , and at 2 h, 1, and 2 days after the end of infusion (T2-4).After MWT was measured at T4, the expression of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in spinal cord dorsal horns was measured by Western blot.Results Compared with group C, the MWT was significantly decreased at T2-4 in RI, E, NE and UE groups, and the expression of p-ERK1/2 was up-regulated in RI, E and NE groups (P＜0.05).Compared with group RI, the MWT was significantly increased at T2-4 , and the expression of p-ERK1/2 was down-regulated in E and UE groups (P＜0.05);and no significant change was found in the parameters mentioned above in group NE (P＞0.05).Compared with group E, the MWT was significantly increased at T2-4, and the expression of p-ERK 1/2 was down-regulated in group UE (P＜0.05).Conclusion The mechaism by which EA at Zusanli reduces hyperalgesia induced by remifentanil in rats with incisional pain is related to inhibited activation of ERK signaling pathway in the spinal cord.