Objective To explore if keratinocytes that stably maintain HPV11 genome can be obtained by transfection and selection methods. Methods Escherichia coil containing pBR322.HPV11 plasmid was cultured and amplified. Then the plasmid was extracted, purified and digested with BamH Ⅰ enzyme to release viral genome from the bacterial vector. After recovering from the low-melting point agarose gel by electrophoresis, the genome was self-circulated with T4 DNA ligase. The religated DNA was cotransfected with pTK-neo DNA into HaCaT keratinocytes using Lipofectamine reagent. After selection with G418 for 2 to 3 weeks, clonal and pooled cultures were expanded and analyzed. Fluorescent quantitative PCR (FQ-PCR) and nested reverse transcriptase PCR (nRT-PCR) were applied to detect HPV11 DNA and spliced HPV11 E1^E4 mRNA expression in the transfected cells. Results After the cotransfection of HPV11 genome into HaCaT keratinocytes and two-week selection,G418-resistant cell colonies were obtained with morphological features indistinguishable from normal HaCaT keratinocytes. As shown by FQ-PCR, HPV11 DNA was present in G418-selected HaCaT keratinocytes. The average viral DNA load capacity was 15.9±16.8 copies/cell in the primary culture of G418-selected HaCaT cells and 23.9±1.1 copies/cell in the third passage of the cells; there was no statistical difference between the two passages of cells (t=-0.822, P>0.05). nRT-PCR targeting HPV11 E1^E4 mRNA transcript produced a specific 628-bp fragment, which was shown by agarose gel electrophoresis. Conclusions Our data indicate that HPV11 genome can be successfully introduced into HaCaT keratinocytes by transfection and HPV11 DNA-positive cells can be obtained by G418 selection. Moreover, HPV11 DNA is still present in the third passage of transfected cells.

Objective:To investigate the clinical efficacy and safety of compound carraghenates suppositories in the treatment of wounds after anal fissure operation. Methods:Eighty-eight cases of patients with anal fistula post-operation wounds were diagnosed from June 2011 to June 2014 and randomly divided into the observation group (n=44) and the control group (n=44). The control group was treated with the conventional treatment and the observation group was treated with external compound carraghenates suppositories additionally. The efficacy, average healing time, growing time of granulation tissue and removal time of necrotic tissue, pain disappear-ance time and wound edema disappearance time after the operation and the area of wound surface in the two groups were observed and compared. Results:After the 14-day treatment, the time of wound healing, granulation growth and cure in the observation group was shorter than that in the control group (P<0. 05). In the observation group, the total effective rate (93. 18%) was higher than that in the control group (77. 27%, P<0. 05). After the 7-day and 14-day treatment, the pain score and wound edema score in the observa-tion group were significantly lower than those in the control group (P<0. 05), and the wound area was significantly smaller than that in the control group (P<0. 05). Conclusion:The clinical efficacy of external compound carraghenates suppositories in the treatment of wounds after anal fissure operation is significant, safe and reliable.

Objective To investigate the effect of repeated hyperbaric oxygen preconditioning (HOP) on the expression of hypoxia-inducible factor-1 alpha (HIF-1a) and erythropoietin (EPO) following spinal ischemiareperfusion (I/R) in rats. Methods Forty-eight male SD rats weighing 250-300 g were randomly divided into 3 groups (n = 16 each): sham operation group (S group); I/R group and repeated HOP group (HOP group). The animals in HOP group were pretreated with O2 ( ＞ 98 % ) at 2.5 ATA I h/d for 5 consecutive days at 24 h before spinal cord I/R. The animals were anesthetized with intraperitoneal pentobarbital sodium 35 mg/kg. Spinal cord ischemia was produced by cross-clamping of abdominal aorta distal to renal artery for 20 min in I/R and HOP groups. Hind-limb motor function was assessed and scored st 48 h of reperfusion. The animals were then sacrificed and the L5-7 segment of the spinal cord was removed for determination of the expression of HIF-1 α and EPO mRNA and protein and microscopic examination. Results Compared with group S, the hind-limb motor function scores and the number of normal motor neurons in the anterior horn of the spinal cord were significantly decreased, and the expression of HIF-1α and EPO mRNA and protein was up-regulated in I/R and HOP groups ( P ＜ 0.05). Compared with group I/R, the hind-limb motor function scores and the number of normal motor neurons in the anterior horn of the spinal cord were significantly increased, and the expression of HIF-1α and EPO mRNA and protein was up-regulated in group HOP ( P ＜ 0.05). The spinal cord injury was attenuated in group HOP compared with group I/R. Conclusion Repeated HOP can reduce the spinal cord I/R injury though up-regulating the expression of HIF1α and EPO in rats.

Objective To investigate the therapeutic effects of repeated transcranial magnetic stimulation (rTMS) at high and low frequency for the rigidity and tremors of Parkinson's disease (PD).Methods Sixty-nine patients with PD received rTMS at 5 Hz or 0.5 Hz.The efficacy was assessed using the unified Parkinson's disease rating scale (UPDRS),motor function tests and motor evoked potentials (MEPs).Results Thirty-two patients with rigidity from PD received 5 Hz rTMS therapy,and their UPDRS total scores decreased significantly.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Thirtyseven patients with tremors from PD received 0.5 Hz rTMS therapy,and their UPDRS total scores were significantly decreased.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Conclusion High frequency rTMS may improve the symptoms of patients with PD rigidity and low frequency rTMS may improve the symptoms of patients with PD tremors.

Objective To discuss the diagnostic performance of MSCT in cystic nephroma(CN).Methods The MSCT findings of CN in 7 patients proved by operation and pathology were retrospectively reviewed.Results All of 7 cases were isolated cystic lesion, including 5 males and 2 females.MSCT scanning showed cystic tumor in the renal parenchyma with well-defined boundary.All cases showed regular cystic wall and septa with no attached cystic wall nodule.The cystic wall and septa appeared homogeneous enhancement on contrast-enhanced CT,while the cystic space showed inhomogeneous density and lack of enhancement.Conclusion The MSCT findings of CN may benefit the definite diagnosis.

Objective To investigate the teaching effect of the PCMC-type (problem-originated clinical medical curriculum) pathology experiment course and clinical pathological conference (CPC) by using microscope digital system.Methods 384 undergraduates of second grade in clinical medicine major and imaging diagnosis major were chosen for the teaching reform project,all of whom were divided into experimental group (192) and control group (192),randomly,and they were taught by PCMC pathological experiment teaching and the traditional pathological experiment teaching separately.After the curriculum,statistic analysis of test score was used to analyze the teaching effect.SPSS 13.0 software diagram method was used to perform Levene variance analysis and t test to the achievement of two groups of students and the students' ability of self evaluation was investigated through questionnaire.Results The average score of the students in the experimental group (86.16 ± 3.28) in the theory exam was significantly higher than that of control group (75.63 ± 2.24) (P=0.000).And the average score of the students in the experimental group in experiment (27.10 ± 0.61) was significantly higher than that in control group (19.87 ± 0.25) (P=0.000).The questionnaire showed that compared with control group,the comprehensive ability of the students in the experimental group was obviously enhanced.Conclusion The PCMC pathology teaching is beneficial to improving the students' ability of linking theory with practice and enhancing their ability to analyze and solve problems,and obviously stimulate their interest in learning.It is worth promoting.

Objective To assess the effects of radiofrequency ablation (RFA) and bipolar cord coagulation (BCC) on the prognosis and complication rate of complicated monochorionic twins.Methods A retrospective review was undertaken in 58 cases of complicated monochorionic twins treated with RFA or BCC at the First Affiliated Hospital of Sun Yat Sen University,from January 2008 to August 2013.Non-parametric Wilcoxon test,Chi-square test,Fisher exact test or multi-variant Logistic regression analysis were used for statistical analysis.Results Indications for selective termination in the 58 cases were:twin reversed arterial perfusion sequence in 12,severe twin to twin transfusion syndrome in 28,discordance of fetal anomalies in 10,selective intrauterine growth restriction in 7 and twin anemia-polycythemia sequence in 1.Forty-three cases were managed with BCC and 15 with RFA.Preterm labor was more common in the BCC group than in the RFA group [86.0％(37/43) vs 9/15,respectively; x2=4.598,P=0.032).Premature rupture of the membranes occurred in 48.8％(27/43) of the BCC group vs.4/15 of the RFA group (x2=2.229,P=0.135).The median procedure-todelivery time was 48 (1-150) days for the BCC group vs.101(14-138) days for the RFA group (Z=-2.245,P=0.025).Overall survival rate was 62.8％(27/43) in the BCC group vs 11/15 in the RFA group (x2=0.547,P=0.460),which was not significantly different.Neurodevelopmental delay was detected in two neonates in BCC group and in one neonate in RFA group.Logistic regression analysis showed that delivery before 28 gestational weeks was an independent risk factor for the poor prognosis of the co-twins (OR=192.720,95％CI:18.610-994.000,P ＜ 0.01).Conclusion Compared with BCC,RFA does not improve the prognosis of complicated monochorionic twins significantly.

Objective To explore the relationship of plasma salusin-? level with the stability,severity,and other risks of coronary atherosclerosis.Methods The patient group included 122 hospitalized patients with coronary artery disease(CAD),whose diagnoses were confirmed by coronary angiography(CAG).The CAD group was further divided into subgroups according to the clinical types,the number of diseased coronary branches,and Gensini's scores.Control group inlcuded 60 healthy subjects who underwent physical examination in our hospital.Salusin-? level and other general biochemical indicators were determined,and the general clinical data were obtained before CAG in all subjects.Results The peripheral blood salusin-? level in CAD patients was significantly lower than that in the controls([0.50?0.18]ng/ml vs [0.69?0.23 ng/ml],P

Objective To investigate the effects of intravitreous injection of conbercept for macular edema secondary to retinalvein occlusion(RVO) during 6 months period.Methods A retrospective clinical study.34 patients (34 eyes) were included in this study,who were diagnosed with macular edema due to retinal vein occlusion by ophthalmologic examination,fundus photography,optical coherence tomography (OCT),fundus fluorescein angiography and other methods.The best corrected visual acuity (BCVA) was examined using the international standard visual acuity chart,and the results were converted to the logMAR visual acuity.The average logMAR BCVA was 0.90 ± 0.68,and the mean macular central retinal thickness (CMT) was (672.27±227.51) μm before treatment.All subjects received intravitreal injection of 0.5 mg conbercept (0.05 ml) at the first visit.Injections were repeated based on the visual acuity changes and the OCT findings.34 eyes received 69 times of injection,the average number of injections was 2.03 ± 1.03.BCVA,OCT were examined before and after treatment using the same method.BCVA and CMT changes,drugs and treatments associated cardiac and cerebral vascular accident,intraocular pressure elevation,retinal tears,retinal detachment,endophthalmitis and other complications after treatment were observed.Linear correlation analysis was used to analyze the correlation between prognosis BCVA and baseline BCVA,correlation between prognosis BCVA and baseline CMT,and also correlation between BCVA and CMT at different time points before and after treatment.Results At 1 week and 1,2,3,6 months after treatment,the average logMAR BCVA was 0.65±0.61,0.56±0.61,0.46±0.55,0.56±0.71,0.44±0.48 respectively.During 1,2,3,6 months after treatment,the mean logMAR BCVA were improved with statistically significant difference (Z=34.029,47.294,41.338,43.603;P＜0.05),while 1 week after treatment showed no obvious improvement (Z=21.941,P＞0.05).At 1 week and 1,2,3,6 months after treatment,the average CMT was (285.89 ± 96.69),(256.65 ± 143.39),(278.68 ± 156.92),(290.11 ± 188.17),(217.15 ± 48.04) μm respectively.At 1 week and 1,2,3,6 months after treatment,the mean CMT were all decreased with statistically significant difference (Z=68.500,98.735,93.235,91.132,109.162;P＜0.05).There was a positive correlation between the prognosis visual acuity and preoperative visual acuity (r=0.682,P＜0.05).However,there was no correlation between the prognosis vision and the degree of macular edema before treatment (r=0.078,P＞0.05).Before and 3,6 months after treatment,BCVA was negatively correlated with CMT (r=0.491,0.416,0.386;P＜0.05),while there was no correlation in other time points (r =0.145,0.217,0.177;P＞ 0.05).Systemic adverse reactions and persistent intraocular pressure elevation,iatrogenic cataract,retinal detachment,retinal tear,endophthalmitis and ocular complications were never found in the follow-up period.Conclusion Intravitreal conbercept is a safe and effective approach for RVO,which can significantly improve visual acuity and reduce CMT.

Objectives To compare total laparoscopic gastrectomy with intracorporeal hand-sewn Gl reconstruction and laparoscopy-assisted gastrectomy for gastric cancer. Methods Between July 2009 and July 2010, 21 patients of gastric cancer underwent total laparoscopic D2 radical gastrectomy with intracorporeal hand-sewn reconstruction and 28 did laparoscopy-assisted D2 radical gastrectomy in Xijing Hospital of Digestive Diseases. All patients were operated on by an experienced surgeon. Patient demographics, TNM stage, location of tumor, the intraoperative and postoperative details of the two groups were compared. Results In the 21 patients undergoing total laparoscopic gastrectomy, there were 15 of distal gastrectomy and 6 of total gastrectomy, compared with 21 and 7 in laparoscopy-assisted group. In total laparoscopic group, intracorporeal hand-sewn technique was used for gastro-jejunal and jejuno-jejunal (J-J)anastomosis, and 25 mm circular stapler was used for esophago-jejunal anastomosis. The operation time was significant longer in total laparoscopic group than in laparoscopy-assisted group of (279 ± 65 ) min vs.(232 ±40) min (P ＜ 0.05 ). No significant difference was observed between the two groups in proximal margin [(5.7 ± 1.5 )cm vs. (5.1 ± 1.4) cm, P ＞ 0.05] and distal margin [( 3.1 ± 0.9 )cm vs. ( 2.9 ±0.9) cm,P ＞0.05]. The iv narcotic use in laparoscopy-assisted group was 1.8 d but it was not used in total laparoscopic group. The first passing flatus was on day 3 in total laparoscopic group compared with 4.8 d in laparoscopy-assisted group. Both groups had 2 postoperative early complications, one intra-abdominal infection and one lung infection in total laparoscopic group compared with one wound infection and one lung infection in laparoscopy-assisted group. There was no anastomosis-related complications after 4 months of follow-up. Conclusions The operation time and postoperative early complication was acceptable for selected patients treated by total laparoscopic D2 radical gastrectomy with intracorporeal hand-sewn GI tract reconstruction in hands of experienced laparoscopic surgeon.