Objective To construct miR-223 knockdown lentivirus vector and provide a tool for further study of the function of miR-223. Methods According to the Invitrogen miR-RNAi online design tool, a pair of complementary oligo?nucleotides encoding miR-223 mature sequence was designed, annealed and ligated with pcDNA6.2-GW/EmGFP-miR vec?tor. Then miR-RNAi expression cassette was cut and subcloned into lentiviral pCDH-CMV-MCS-EF1-copGFP vector. The lentiviruses were packaged and titered, and then ST2 cells were infected with viruses. The efficiency of infection was calcu?lated, and the knockdown of endogenous miR-223 was detected by using real-time RT-PCR. Results Restriction enzyme digestion and sequencing results showed that miR-223 lentivirus construct was successfully made. Lentivirus that knock?down miR-223 expression packaged and infected of target cells. The expression of GFP green fluorescent protein accounted for 80%-90%and the virus titer was 1×109 PFU/mL. The infection efficiency reached 90%. Compared with negative control virus, miR-223 knockdown lentivirus significantly down-regulated the expression of miR-223 in ST2, and was 31%(n=3, t=15.091, P<0.05). Conclusion miR-223 knockdown lentivirus is successfully made. It provides a tool for further studying the function of miR-223.

Objective: To analyze the diagnostic methods and risk factors of female lung cancer, and to investigate the role of flberoptic bronchoscopy in the early diagnosis of lung cancer. Methods: The bronchoscopy data of 238 female lung cancer patients seen in our hospital between January 2005 and December 2008 were enrolled. The fiberoptic bronchoscop-ic features with clinical imaging, main risk factors of lung cancer and the pathological types were analyzed. χ~2 test was used for statistical analysis. Results: The major pathological type of lung cancer was adenocarcinoma in females younger than 40 years and squamous cell carcinoma in patients older than 60 years. The proliferative type was correlated with squa-mous cell carcinoma. Its imaging manifestion was a mass and the main risk factor was smoking. The infiltrating type was correlated with adenocarcinoma. Its imaging manifestation was mainly pleural effusion and the main risk factor was indoor soot inhalation. Conclusion: Females exposed to indoor soot inhalation for a long time especially those with a number of risk factors should be offered a screening with low dose CT (LDCT) regularly. Fiberoptic bronchoscopy and cytohistology are of great value for early diagnosis of lung cancer in those with abnormal findings in CT.

Objective To analyze the pathological diagnosis of patients with clinically diagnosed eczema,to describe the spectrum of skin diseases tending to be misdiagnosed as eczema,and to investigate factors associated with their misdiagnosis.Methods A retrospective study was performed on 400 patients who were clinically diagnosed as eczema and received pathological examination at the dermatology clinic of Peking University Third Hospital from August 2006 to April 2013.Skin biopsy specimens were re-reviewed for these patients,and pathological diagnosis was made in combination with clinical presentations.Results Of the 400 outpatients with clinically diagnosed eczema,110 (27.5％) were finally diagnosed as non-eczema skin diseases pathologically,including 16 cases of psoriasis,13 bullous pemphigoid (BP),11 lichen planus (LP),9 cutaneous amyloidosis,8 mycosis fungoides (MF),14 skin malignancies,and 39 other skin diseases.The highest misdiagnosis rate was observed in people aged 60 to 79 years (33.9％) and lesions at the genital sites (46.2％).Conclusions Many conditions tend to be misdiagnosed as eczema in clinic,including psoriasis,BP,LP,cutaneous amyloidosis,MF and skin malignancies.Misdiagnosis is rather frequent in elderly people and eczematous lesions in genital areas,and pathological examination should be taken actively for uncertain cases.

Objective: To investigate the inhibitory effects of RNA silencing via adenovirus-mediated vascular endothelial growth factor receptor (VEGFR) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo. Methods: Ad-VEGFRshRNA adenovirus containing enhanced green fluorescent protein (EGFP) gene and VEGFRshRNA was constructed and was used to infect A549 cells; fluorescent microscopy was used to observe the infection efficiency. Western blotting assay was used to examine the expression of VEGFR protein in A549 cells. MTT method was used to examine the cell viability and the cell growth curve was drawn. The inhibition of cell growth was examined by cell cycle and colony-forming test. Meanwhile, nude mice were transplanted with A549 cells to establish tumor-bearing model, and the long term growth of tumor was observed. Results: Western blotting revealed that the expression of VEGFR was obviously decreased in the RNA interference group. The cell growth curve indicated that the cell growth was obviously inhibited after RNA interference. Cell cycle and colony-forming test indicated that the tumor growth was obviously inhibited after RNA interference. In vivo study with nude mice also indicated that RNA interference obviously inhibited tumor growth. Conclusion: The constructed VEGFR-targeted shRNA can effectively inhibit VEGFR expression in A549 cells and can suppress the growth of A549 cells.

Objective:To investigate the role of platelet-derived growth factor(PDGF)-AB on angiogenensis in aged mice.Methods:Cardiac microvascular endothelial cells(CMEC)from the hearts of young adult and old mice were cultured.Expression of PDGF-BB and PDGF-AA was detected by RT-PCR.The migration of CMEC was determined with ChemoTX Chamber.Ear angiogenesis model was made in mice.Blood flow in neo-microvessels and collateral vessels was measured with Laser Doppler.Then biotin-labeled Dextran-lysine was injected into the mice through cardiac puncture to label vessels.Ears were cut and immunohistochemistry was carried out by ABC method.Results:Both PDGF-BB and PDGF-AA were highly ex-pressed in CMEC of young adult mice;expression of PDGF-BB was not detected in aged mice.PDGF-AB reverted the levels of PDGF-BB in aged CMEC to the levels of young adult mice.Migration rate of CMEC in aged mice was significantly increased after stimulation by PDGF-AB(P

ObjectiveTo assess malignant trophoblastic neoplasia with the standards of the clinical stage and prognostic factor scoring system. MethodsThrough assessing the high-risk factors except clinical stages for 223 patients before treatment according to International Federation of Gynecology and Obstetrics(FIGO) scoring system published in 2000, appropriate treatments were selected for the different patients. ResultsForty-three of 78 cases of choriocarcinomas were with high-risk factors, the other 35 cases were with low-risk factors; 7 of 145 cases of invasive moles were with high-risk factors and the others were with low-risk factors. The primary chemotherapy principle was that one agent was used for those patients with low-risk factors and two or multiple-agents were used for those patients with high-risk factors. Among all patients, the one-year, three-year and five-year survival rates were 98.6%,98.1% and 97.1% respectively. No patient died of drug toxicity or complication. ConclusionSelection of treatment approaches according to the prognostic assessment of malignant trophoblastic neoplasia could lead to promising survival rate with no uncurable complication and toxic effects.

Percutaneous endoscopic lumbar discectomy belongs to minimally invasive spine operation Its superiority includes smalleroperation wound,less bleeding,shorter hospital day,and earlier return to function,conpared with the traditional operation.At the same time,percutaneous endoscopic lumbar discectomy has complications,as the open operation.This paper reviews its common complications,diagnosis,prevention and control.

OBJECTIVE: To investigate the stability of 0.5% glutaraldehyde solution. METHODS: Classic constant temperature accelerated test method and sample observation method were applied with the content of glutaraldehyde as index. The content of glutaraldehyde was determined by titration which was determined by titration. RESULTS: The content change of 0.5% glutaraldehyde solution was in line with the first order kinetics, and the results of both methods were similar. The validity duration of 0.5% glutaraldehyde solution was 85.7 days at room temperature (25 ℃). CONCLUSION: 0.5% glutaraldehyde solution should be used up as soon as possible and the perfect time is less than 2 week.

Objective To study roles of GRP78 and GRP94 in tumorigenesis and progression of colorectal carcinomas (CRC).Methods Colorectal carcinoma tissues of 124 cases were studied for GRP78 and GRP94 proteins by immunohistochemistry and Western blotting.The correlations among the expression of proteins and clinicopathologic parameters and overall survival of CRC were analyzed using univariate and Cox regression analyses.Results The overexpression of GRP78 and GRP94 was detected in the cytoplasms or plasma membrane of the tumor cells.They were positively correlated with tumor differentiation,staging,lymph node metastasis and remote metastasis.Survival analysis showed that overexpression of both proteins was associated with poor outcome of CRC ( P =0.025,P =0.010,respectively).Cox regression analysis demonstrated that the expression of GRP94 was an independent poor prognostic factor for CRC.Conclusions Up-regulated expression of GRP78 and GRP94 was possibly involved in the pathogenesis,growth,invasion and metastasis of colorectal carcinomas.GRP78 and GRP94could be seen as a marker for the aggressive behavior and poor prognosis in colorectal carcinomas.

0.05 ). But there was significant difference in islet function when warm ischemia time was prolonged to 30 min compared with that in site perfusion group ( P