Objective To construct an eukaryotic expression vetor of ring finger protein 6 (RNF6)and to investigate its ubiqu-itylation on insulin receptor substrate-1(IRS-1).Methods Human RNF6 coding sequence was amplified by polymerase chain reac-tion (PCR)method with human cDNA as template.The pcDNA3.1-CHA-RNF6 was constructed with routine molecular methods and transfected into hepatocarcinoma cell HepG2.Relative quantification of IRS-1 mRNA was preformed by real-time reverse tran-scription PCR.Western blot was applied to detect the expression levels of IRS-1.Results 48 h after the plasmid carring RNF6 gene transfected HepG2 cells,the mRNA level of ISR-1 gene reduced to 69% of control group.The expression of IRS-1 was markedly lower than control group (P <0.01).Conclusion The expression of IRS-1 is markedly down-regulated in HepG2 and enhancement of the ubiquitylation level of IRS-1 contribute to the disorder in insulin signal transferring.

Objective To observe the effects of Bushen Huoxue Decoction on clinical symptom, vertigo degree and cognitive function for elderly patients with chronic cerebral circulation insufficiency (CCCI); To explore its possible mechanism. Methods Totally 70 cases were randomly divided into observation group and control group according to the random digits table method, with 35 cases in each group. The two groups were given Western medicine consisting of blood pressure control, glucose control and blood lipid control. Based on this, the control group was given Flunarizine Hydrochloride capsules, 5 mg once a day orally before sleeping. The observation group was given Bushen Huoxue Decoction, one dose a day and taken twice a day. The treatment lasted for 30 days. The TCM symptom score, Chinese version of dizziness handicap inventory (DHI) and Montreal cognitive assessment (MoCA) score were observed. The mean blood flow velocity (Vm) of basilar artery (BA), bilateral vertebral artery (VA), bilateral middle cerebral artery (MCA) and the content of plasma acid phospholipid (AP) were detected, and the clinical efficacy was evaluated. Results The total effective rate of the observation group was 94.3％ (33/35), which was significantly higher than that of 82.9％ (29/35) in the control group, with statistical significance (P<0.05); after treatment, the TCM symptom score, DHI score, and the content of plasma AP were significantly lower than those of pre-treatment in the same group (P<0.05); the MoCA score and the Vm of VA, LBA, RBA, LMCA, RMCV were significantly higher than those of pre-treatment in the same group (P<0.05); the TCM symptom score, DHI score, MoCA score, the Vm of LVA, LMCA, RMCA and the content of plasma AP in the observation group were improved better than those in the control group, with statistical significance (P<0.05). Conclusion Bushen Huoxue Decoction has an obvious therapeutic effects for elderly patients with CCCI, which can improve clinical symptom, vertigo degree and cognitive level, which possible mechanism is related to increasing the cerebral blood flow velocity and reducing the content of plasma AP.

Objective To investigate the significanceof both the adenomatous polyposis coli (APC)and deleted in colorectal carcinoma (DCC)gene methylation in the early diagnosis of lung cancer.Methods 245 patients with lung cancer and 150 patients with non-malignant lung disease patients and 40 healthy volunteers were drawn for the experiment.A methylation specific PCR (MSP)was used to detect the methylation status of APC and DCC in the peripheral blood.Results The posi-tive rates of APC and DCC genes promoter methylation Peripheral blood of patients with lung cancer were 26.53% (65/245)and 36.33% (89/245),respectively.The positive rates of APC and DCC genes promoter methylation Peripheral blood of patients with benign lung diseases were 2.67% (4/150),8.00%(12/150),respectively.The positive rates of APC and DCC genes promoter methylation Peripheral blood of healthy volunteers were 0.There was a significant difference between patients with the lung cancer,those with the benign lung diseases and Healthy volunteers (P <0.01).The sensitivity and specificity of lung cancer was respectively 52.65% and 89.33% by the combination of APC and DCC genes methylation. Comparing with the separate APC detection,sensitivity and specificity statistical differences were significant in joint meas-urement (P <0.01).Comparing with the separate DCC detection,sensitivity statistical differences were significant,but spe-cificity statistical differences was not obvious (P >0.05).There was no correlation between methylation test results and pa-tient gender,age,pathological type,pathological grade,TNM stage and so on (P >0.05).Conclusion APC and DCC gene methylation were closely related to the development of non-small cell lung cancer.They can be used as an early diagnostic marker of lung cancer.

Objective To evaluate the efficacy, safety and complications of minimally invasive percutaneous radiofrequency ablation(PRFA) therapy for hepatic cavernous hemangioma(HCH). Methods Twelve patients with total 15 HCHs (2.5cm to 9.5cm) were treated by using RF-2000 generator and 10-needle LeVeen electrode percutaneously guided by B-ultrasound. Tumors larger than 3cm were treated by multiple overlapping ablations that encompassed the entire lesions as well as an approximately 0.5cm rim of normal liver tissue. Results The procedures were successful and the patients recovered well without the complications of bleeding or bile leakage after mean 48-month follow up period. Spiral CT scanning showed that the contrast enhancement was disappeared within the lesions or at their periphery in all cases. The mean diameter of the ablated HCHs reduced 68%. Conclusion The results suggested that PRFA therapy was a minimally invasive, simple, safe, and effective approach for treating HCHs in selected patients.

Objective To investigate the urinary iodine level and the thyroid function of early pregnant women in Qingdao,to explore the relationship between the iodine nutritional status and hypothyroidism.Methods 326 early pregnant women (<12 gestational weeks)were collected.The urinary iodine concentration and serum thyroid stimulating hormone (TSH),free thyroxine (FT4 ),thyroid peroxidase antibody (TPOAb)were detected. Results The median of urine iodine of pregnant women was 176.57μg/L,and the ratios of iodine deficient,iodine adequate,iodine more than adequate and iodine excessive groups were 3 8.65%,41.10%,15.95% and 4.30%.The total ratio of innormal urine iodine was 58.9%(192 /326).The differences between the ratios of the iodine deficient group,iodine excessive group and iodine adequate group both had statistically significant(χ2 =4.308,P <0.05;χ2 =4.826,P <0.05 ).There was no effect of iodine intake levels on serum thyroid peroxidase antibody.Conclusion There is no iodine deficiency among early pregnant women in Qingdao,but still individuals expose iodine deficiency.Iodine deficiency and iodine excessive are the risk factors of hypothyroidism during early pregnancy women,so we commend to detect the urinary iodine level.

3.0 cm in diameter were given multiple overlapping ablations that encompassed the entire lesions as well as an approximately 0.5 cm rim of normal liver tissue. The operation was performed under local anesthesia in association with systemic analgesia. Results The procedure was successfully accomplished in all the 26 patients. Adverse reactions happened at 2~5 postoperative days, including local pain and discomfort, pyrexia, and elevations of ALT levels. No bile leakage or hemorrhage occurred. Follow-up observations in 23 patients for 1~5 years (mean, 2 years and 8 months) showed a complete response (CR) in 22 patients and a partial response (PR) in 1 patient by one mission of PRFA, the effective rate (CR+PR) being 100%. The hepatic cavernous hemangiomas decreased by a mean of 68.8% (range, 41.2%~81.5%) in diameter. Conclusions Minimally invasive PRFA is a safe and effective therapy for HCHs in properly selected patients by experienced hands.

Objective To construct an objective,comprehensive core competence evaluation index system for nursing postgraduates,and to provide evidence for postgraduates nursing education.Methods According to literature review,expert interviews,focus group discussions,Delphi and other research methods,we conducted two rounds of 32 experts consultation in 18 cities from 10 provinces to establish the final core competence evaluation index system.Results The two experts consultation derived core competence evaluation index system including 7 level-1 indexes,20 level-2 indexes and 53 entries.Level-1 indexes:theoretical knowledge and skills,critical thinking ability,research ability,clinical care ability,teaching skills,interpersonal communication skills,professional development skills.Conclusions This program is rigorous and reliable,with some theoretical and practical significance of its results,which provide a reference for the training program,evaluation of the quality,assessment of postgraduate nursing education.

Objective To investigate the risk factors of severe intrahepatic cholestasis during early period after liver transplantation.Methods The clinical data of 225 patients who received orthotopic liver transplantation at the Nanfang Hospital of Southern Medical University from August 2004 to February 2011 were retrospectively analyzed.All patients were divided into positive group (60 patients with intrahepatic cholestasis) and negative group (165 patients without intrahepatic cholestasis).Preoperative,intraoperative and postoperative factors of the 2 groups were compared via t test,chi-square test,Wilcoxon test or Logistic regression analysis.Results The proportion of patients with hepatic cirrhosis,hepatic encephalopathy integral,ascites integral,international normalized ratio,and the levels of prothrombin time (PT),total bilirubin (TBil),aspartate aminotransferase of the positive group before operation were significantly higher than those in the negative group (x2 =6.09,Z =2.22,2.60,2.46,2.84,4.81,3.42,P ＜ 0.05),while the levels of albumin,Na +,K +,hemoglobin,platelet (PLT) of the positive group in the operation were significantly higher than those in the negative group (t =2.10,4.97,Z =2.49,t =3.51,Z =3.66,P ＜ 0.05).The ratio of compatible blood type of the donors and recipients,ratio of fatty liver graft,cold ischemia time,relative warm ischemia time,intraoperative blood loss,intraoperative transfusion of red blood cells,PLT,and cryoprecipitate of the positive group after the operation were significantly higher than those in the negative group (x2 =4.29,13.11,Z =2.45,2.61,3.75,3.20,2.89,3.95,P ＜0.05).The incidences of acute rejection,hepatic artery embolism,pulmonary infection,bacteraemia,fungal infection and cytomegalovirus (CMV) infection were significantly higher than those in the negative group (x2 =9.87,4.91,8.21,6.29,3.92,9.26,P ＜0.05).The results of multivariate analysis revealed that preoperative level of TBil ＞ 51.3 μmol/L,fatty of the liver graft,intraoperative transfusion of cryoprecipitate,postoperative acute rejection,hepatic artery embolism,postoperative pulmonary infection,bacteraemia,CMV infection were independent risk factors of severe inrahepatic cholestasis (OR =15.82,7.99,2.88,3.03,53.20,3.34,4.11,3.22,P ＜ 0.05).The incidence of severe intrahepatic cholestasis was significantly lower in patients with higher level of PLT and longer PT (OR =0.33,0.25,P ＜ 0.05).The mortality rates of the positive group and negative group at 6 months after the operation were 41.7％ (25/60) and 19.4％ (32/165),and the mortality rate of the positive group was significantly higher (x2 =11.54,P ＜ 0.05).Conclusion Correction of poor clinical status before liver transplantation,reinforcement of infection control and anti-rejection may reduce the incidences of complications and decrease the associated early mortality.

Objective To report a case of chromomycosis caused by Phialophora verrucosa and explore the laboratory features of the pathogen. Methods Skin lesion was examined by histopathology and fungus culture. The morphology of the isolate was observed by microscopy and scanning electron microscopy. The coenzyme Q system of this isolate was analyzed by HPLC assay. The DNA sequences of LSU rDNA D1/D2 region of this isolate and a standard fungus strain were compared. Results The initial lesion was an erythematous papule that subsequently developed into one or multiple coalescing warty papules or plaques slowly. The bronze-colored spores could were observed in the dermis or macrophages. The isolate grew very slowly, requiring 4 weeks of incubation. Microscopically, no characteristic structures were found on Sabourand′s dextrose agar, while there were vase-like structures, which were referred to as phialides on potato dextrose agar (PDA) and corn meal agar I (CMA-I). The phialides on PDA mostly grew at the top of hypha, but on CMA-I they mostly grew on the side of hypha. The isolate contained coenzyme Q-10, and its DNA sequence of LSU rDNA D1/D2 region completely consistent with those of the standard strain. Conclusion Chromomycosis caused by Phialophora verrucosa is rare in China. It can be diagnosed by fungus culture and histopathological examination. Coenzyme Q system analysis and DNA sequencing can exclude the interference from different phenotypes.

Objective To observe the significance of tumor abnormal protein (TAP)in the therapeutic monitoring of non-small cell lung cancer (NSCLC).Methods Peripheral blood from 30 NSCLC patients were collected to make smears at the moment of pre-treatment,half a month,one month,3 months and 6 months after therapy.TAP was detected by coacervation method.The maximum area of condense was applied to estimate the level of TAP.Thirty healthy subj ects were chose as con-trol group.Results The positive rate of TAP in NSCLC patients was 86.67%,and 3.33% of healthy subjects were positive in TAP.The difference was statistically significant (χ2=140.3,P<0.01).The condense area of TAP in patients withⅢ~Ⅳ stage of NSCLC [411(89,562)mm2]was significantly higher than those withⅠ~Ⅱ stage NSCLC [267(31,407)mm2]. The condense areas in both of two groups went down after treatment.A significant difference of condense area appeared inⅠ~Ⅱ stage of NSCLC patients a month after therapy as well as Ⅲ~Ⅳ stage of NSCLC patients half a month after treat-ment.The condense areas went to their lowest level 3 months after therapy.ForⅠ~Ⅱ stage patients,the condense area fell to 21% compared to that before treatment,but for patients withⅢ~Ⅳ stage of NSCLC,37% of pre-treatment condense ar-ea were detected.TheⅠ~Ⅱ stage of NSCLC patients had a greater decrease in condense area of TAP than theⅢ~Ⅳ stage patients by 3 months after treatment (χ2=6.22,P<0.05).Conclusion Detection of TAP in peripheral blood had a high sensitivity for NSCLC,and is able to monitoring the treatment effect.