In order to study the pathogenesis of HPVl1 and seek for a therapeutic approach of the disease caused by HPV1l. The HPVl1/E2 644bp was amplified by PCR with HPVll plasmid DNA. pGEM-T Easy was used as vector and a clone pTV-644 was obtained.The inserted DNA sequencing was carried out after selecting and identification. According to the hammerhead structure described by Symon's, the possible secondery cleavage sites on HPVll/E2 mRNA were analyzed and the secondery structure of substrate was predicted by computer and ribozyme, excluding analogous sequence of substrate combined with ribozyme were found in the mRNA.The hammerhead ribozyme of RZ2777 against HPV1l/E2 mRNA was seleted to carry out cleavage reaction in vitro. Results of the experiment showed that 644bp substrate derived from HPVll/E2 can be cleavaed site-specifically by ribozyme in vitro. The cleavage activity showed over 85% by choosing the optimun reaction condition, which was not affected by two cis-ribozymes on both 3′-and 5′-ends released by self cleavage,but two flan-king sequences of target RNA influenced the cleavage activity. Results demonstrated that the ribozyme will become a highly effective and specific therapy against HPVll infection.

Objective The aim of this study was to determine if isoflavone genistien has protective effects against high glucose-induced cell apoptosis in human aortic endlthelial cells,and investigate the possible mechanism for this protection.Methods Human aortic endothelial cells subjected to normal (5mmol/L) or high glucose (25mmol/L) were treated with genistein at 0,50,100nmol/L.Parallel experiments were performed with 100nM 17b-estradiol,and also in the presence and absence of the pure anti-estrogen ICI-182,780 (100nmol/L).The effects on cell apoptotic DNA fragmentation were determined using cell death ELISA,and the effects on cellular proliferation were determined using tritiated thymidine incorporation assay.Estrogen receptor expression was detected by Taqman quantitative PCR.Results Genistein at 100nmol/L significantly reduced high glucose-induced DNA fragmentation,and reversed cell DNA synthesis inhibition (P＜0.001) after 24 hours' incubation.The effect of genistein was completely blocked by ICI-182,780administration.Estrogen receptor beta,but not alpha was found to be expressed in these cells.Conclusion Isoflavone genistein shows protection against high glucose-induced cell damage through estrogen receptor beta,reducing apoptotic DNA damage and protecting from the inhibition of cell proliferation.

OBJECTIVETo observe the clinical effect and safety of circumcision stapler in the treatment of phimosis and redundant prepuce.

METHODSWe treated 120 patients with redundant prepuce or phimosis using circumcision stapler and another 60 by conventional dorsal-incision circumcision. We observed intraoperative blood loss, operation time, postoperative pain, wound healing time, cosmetic appearance of the penis, and postoperative complications and compared them between the two groups of patients.

RESULTSStapler circumcision showed obvious advantages over the conventional method in intraoperative blood loss ([2. 3 ± 1. 3] vs [15.6 ± 2.9] ml), operation time ([7.1 ± 1.4] vs [22.6 ± 4.6] min), wound healing time ([12.0 ± 2.9] as [16.3 ± 3. 1] d), postoperative pain score (1. 9 ± 1. 3 vs 5. 2 ± 1. 7), incision edema, and cosmetic appearance of the penis (all P <0. 05). Besides, stapler circumcision exempted the patients from stitch-removal pain. However, the incidence rate of postoperative local ecchymosis was significantly higher in the circumcision stapler group than in the conventional circumcision group (20. 8% vs 8. 3% , P <0. 05).

CONCLUSIONCircumcision stapler, with its advantages of easier manipulation, shorter operation time, better cosmetic penile appearance, less pain, and fewer complications, is superior to conventional circumcision in the treatment of phimosis and redundant prepuce.

Blood Loss, Surgical ; Circumcision, Male ; instrumentation ; methods ; Ecchymosis ; etiology ; Humans ; Male ; Pain, Postoperative ; Penis ; abnormalities ; Phimosis ; therapy ; Postoperative Complications ; Postoperative Period ; Surgical Staplers ; adverse effects ; Wound Healing

Background Researches determined that the alteration of A69S locus of age-related maculopathy susceptibility 2 ( ARMS2 ) gene is closely associated with the pathogenesis and progression of age-related maculopathy ( AM D ).However,the location of ARMS2 protein in normal eye tissue is still in controversy,therefore,its function is below understanding up to now.Objective The goal of this laboratory work was to investigate the distribution,expression and location of ARMS2 protein in normal adult retina and choroid as well as in retinal pigment epithelial (RPE) cells and lay a basis for exploring further its function in the protein level.Methods Ten donor eyeballs of normal adult male with the age from 28-42 years were collected in eye bank of Qingdao Eye Hospital.The frozen sections of the retina and choroid were prepared for the detection and location of ARMS2 in 3 eyes by immunofluorescence under the confocal laser microscope.The retina was isolated for the primary culture of RPE cells using explant culture method.The cells were then identified by CK32 antibody by immunofluorescence.The distribution and expression of the ARMS2 protein in retina,ehoroid and RPE cells were determined by immunofluorescence technique.Results ARMS2 protein was strongly expressed in retinal vessel,RPE cell layer,Bruch membrane and choroidal vessel,but weak expression was in retinal ganglion cell layer,inner nuclear layer,outer plexiform layer,outer nuclear layer and inner plexiform layer in the normal eyes.The primarily cultured cells appeared the polygon shape with the abundant pigment in cytoplasm.The immunofluorescence of the cells showed the positive response for CK32,exhibiting the green fluorescence granules in the cytoplasm.The positive expression of ARMS2 protein also was seen in the cytoplasm of RPE cells,appearing the red fluorescence.Conclusions ARMS2 protein mainly distribute and locate retinal and choroidal vessels,RPE cells and Bruch membrane in normal eye.

Objective To explore a more reasonable and effective therapeutic regimen and evaluate prognostic factors in hepatoblastoma patients after combined therapy.Methods Sixteen patients diagnosed on hepatoblastoma between Jan.2000 and Nov.2007 were reviewed and followed-up.Resection with chemotherapy was taken among 16 cases.Chemotherapy included pre-operation and post-operation.Five cases were cured by transcatheter arterial chemombolization(TACE).Six cases were cured by single chemotherapy(both TACE and single chemotherapy were taken in 2 cases).Five cases weren't cured by pre-operation chemotherapy.Nine cases were subjected to curative resection,3 cases to alleviative resection,2 cases with single chemotherapy,1 case with single TACE,and 1 case refused operation and left hospital.Their mean survival duration was 13.5 months(3-98 months).SPSS 13.0 software was used to analyze the data.Results The total survival rate of cases as stage Ⅳ as lower than cases as stage Ⅰ,Ⅱand Ⅲ.Both alpha-fetoprotein(AFP)100 000 ?g/L cases had worse survival rate.The prognosis of mixed type was better than fetal type,embryonal type and anaplasia type.The survival rate of cases with standard chemotherapy was higher than cases with unstandard chemotherapy.And the surgical resection cases had better survival chance than non-surgical resection cases.The survival rate of surgical residual cases was worse than non-surgical residual cases.Conclusions Chemotherapy can improve the total survival rate and long-term survival rate of hepatoblastoma patients.TACE is a safe and effective choice for hepatoblastoma patients.We need to be alert and make the diagnosis as early as possibe,and treat it early and properly.

Objective To investigate the clinical features,treatment response and prognosis in children with Takayasu′s arteritis(TA) in order to improve the understanding of TA.Methods A retrospective study of 10 children with TA was performed.All of them were admitted and diagnosed in Peking University First Hospital from Jan.1998 to Oct.2008.The clinical features,laboratory tests,imaging modalities,treatment response and prognosis were all collected and evaluated.Results There were 3 boys and 7 girls in the 10 patients with TA,and the ratio of male to female was 12.3.The onset was from 4 months to 9 years old,with average age at 5.5 years old.The average duration of diagnosis was 7.6 months.The incidences of hypertension,vascular bruits,albuminuria,convulsion were present in 100%,100%,70% and 40%,respectively.The clinical types included typeⅡ(60%),type Ⅲ(10%) and type Ⅳ(30%).The acute phase inflammatory indices of activity such as erythrocyte sedimentation rate(ESR),C-reactive protein(CRP) and white blood cell(WBC) were not evidently increased.Tuberculosis infection was found in 6 out of 10 patients and anti-tuberculosis treatment was performed.Six patients were treated with steroids and 3 cases of them were also given immunosuppressives cyclophosphamide or methotrexate.Three of the 10 patients received anti-hypertensive and vasodilator.Two patients received percutaneous translurminal angioplasty and 1 patient received nephrectomy.One patient died of renal failure,heart failure and shock.Conclusions The patients with TA had high prevalence of tuberculosis infection,diagnosis as often late because of lack of specific clinical features at the acute inflammatory period.When organic ischaemia occurred,treatment response was usually unsatisfactory.Patients with multi-systemic and multi-viscera lesions should have comprehensive examination,especially for those with hypertension,pulseless and vascular bruits,in order to rule out TA.Early ultrasonography,computed tomography and magnetic resonnance image methods are valued in eariler diagnosis and they are the key factors to improve prognosis.

Objective To evaluate the myocardial blood supply in patients with metabolic syndrome (MS) using 99Tcm-MIBI SPECT MPI. Methods A total of 342 patients were divided into four groups according to the number of abnormal metabolic indices: no abnormal metabolic index (Group 1), one abnormal index (Group 2), two abnormal indices (Group 3), three or more abnormal indices (Group 4). Each patient underwent two-day protocol of gated stress and rest 99Tcm-MIBI MPI. One hundred and three of the 342 patients were clinically diagnosed as MS and underwent CAG within 1 month after MPI. χ2test was used to evaluate the difference among the four groups and Kappa test to analyze the correlation between MPI and CAG. Results Compared with CAG, the diagnostic sensitivity, specificity, positive and negative predictive values by 99Tcm-MIBI SPECT MPI for coronary artery diseases (CAD) in 103 MS patients were 80.5% (33/41), 85.5% (53/62), 78.6% (33/42) and 86.9% (53/61), respectively. The correlation coefficient between MPI and CAG was 0.657 (P<0.001). The abnormal MPI rates in group 1, 2, 3 and 4 were 23.3% (10/43), 32.9% (26/79), 54.4% (56/103), and 57.3% (67/117), respectively (χ2=23.22, P<0.001). Conclusions In MS patients,99Tcm-MIBI SPECT MPI can be useful for evaluating myocardial blood supply and the myocardial ischemia rates may correlate positively with the number of abnormal metabolic indices.

The effects of surfactants on the production of cellulase by Trichoderma viride in liquid substrate fermentation process were investigated. Straw was used as the sole carbon source and the surfactants were biosurfactant rhamnolipid from Pseudomonas aeruginosa and Tween 80. The changes of FPA,CMCase,Avicelase and surface tension with time were analyzed under different concentrations of the two surfactants. The results showed that the surfactants can enhance the enzyme activity of Trichoderma viride. The FPA,CMCase,Avicelase were promoted 1.08,1.6 and 1.03 times higher than the controls by rhamnolipid. The enhancement of the enzyme activity by rhamnolipid was much higher than that of Tween 80. At the same time,rhamnolipid was not degraded prior to other substrate.

Objective To improve the diagnosis and treatment of ureteral cancer. Methods A retrospective analysis of 24 cases of primary ureteral cancer treated from January 1990 to March 2005 was performed.The diagnostic value of ultrasound,IVU,CT,MRU and the patients' outcomes were reviewed. There were 19 males and 5 females aged 38-72 years(mean,59 years).The tumors were on the left side in 16 cases and on the right in 8.Of the 24 cases,17(71%)had gross hematuria and 7(29%)had micro- scopic hematuria.Urine cytology was performed in 16 cases with a positive rate of 6.3%.B-ultrasonic exami- nation showed hydronephrosis in 19 cases(79%)and low-echo space-occupying disease of middle-inferior ureter in 3(12%).IVU demonstrated hydronephrosis in 20 cases(83%)and filling defect of the diseased ureter in 3(12%).Retrograde pyelography showed filling defect of the diseased ureter in 16(76%)of 21 cases(5 cases had failure of intubation).CT scan was performed in 20 cases,indicating thickening of the ureteral wall and infiltration of the cancer in 14(70%).In 3 cases who had undergone spiral CT thin layer scan and 1 of 3 cases who had undergone MRU,the definite diagnosis was made.Results All the 24 pa- tients underwent surgical treatment.Among them,nephroureterectomy and bladder cuff or partial resection were performed in 18 cases,and nephrectomy and partial ureterectomy in 6 cases.Postoperative pathology showed transitional cell carcinoma in 23 cases,and adenoma in 1.Of the 14 cases during 1990-1999 peri- od,1,5,3,2,2 and 1 cases had survival time of 1,2,3,4,5 and 6 years,respectively.Of the 10 cases during 2000-2005 period,3 were lost to follow-up;2 survived for 3 years and 2,for 1 year;the other 3 who have survived near 5 years have been followed till now.Conclusions IVU and retrograde urography are the most common diagnostic measures for primary ureteral cancer.They can be used in combination with other imaging study to reduce missed diagnosis rate.The 5-year survival rate was lower because of late pathologic stage of the tumors in the patients of this series.

OBJECTIVETo study the local infiltration length of spinal bone giant cell tumor and chordoma in various para-tumorous tissues.

METHODSSurgical specimens from 7 patients with bone giant cell tumor and 10 patients with chordoma were divided into 6 groups: cortical bone group (CBG, para-tumorous tissue is bone cortex with periosteum), cartilage group (CG, para-tumorous tissues contain intervertebral disc, cartilage endplate and facet joint cartilage), trabecular bone group (TBG), paraspinal muscle tissue group (MTG), adipose tissue group (ATG) and scar tissue group (STG). Macroscopically margin (MSM) in the fresh specimens, histological margin (HLM) in the H&E sections and molecular margin (MCM) in the immunohistochemical staining sections were measured respectively. Three types of the margins of each group and MSM-HLMs, and MSM-MCMs among all groups were compared.

RESULTSAnalysis of Variance comparing MSM, HLM and MCM within each group showed that there were significant differences in all groups (P < 0.05) except for CBG and CG. MSMs were significantly longer than HLMs and MCMs in TBG and MTG (P < 0.05), and MSMs were significantly longer than MCMs in ATG and STG (P < 0.05). MSM-HLMs and MSM-MCMs of TBG and ATG were significantly larger than those of CG and CBG. MSM-HLMs and MSM-MCMs of MTG and STG were significantly larger than those of CG; in addition, MSM-MCM of STG was significantly larger than CBG's. The maximum of tumor infiltration length in CBG, MTG, ATG and STG were 11.68 mm, 13.08 mm, 8.64 mm and 9.98 mm respectively.

CONCLUSIONSEach vertebra can be seen as a compartment, so total spondylectomy is an optimal procedure when bone giant cell tumor and chordoma localized in the compartment. To achieve a wide resection, a 1.3 cm wide margin in para-tumorous cancellous bone, a 1.5 cm wide margin in para-tumorous muscle and 1.0 cm wide margin in adipose tissues are necessary. First-time resection should be more aggressive and avoid tumor cell contamination. The surgical extent of salvage revision should contain para-tumorous scar tissue with the margin no less than 1.0 cm.

Adolescent ; Adult ; Aged ; Chordoma ; pathology ; surgery ; Female ; Giant Cell Tumor of Bone ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Spinal Neoplasms ; pathology ; surgery ; Spine ; pathology