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 search for the reasons of high positive rate of amotile bacteria and the diagnosis of septicemia in new-born Methods The blood was drawn from the different site of the new-born with septicemia and carricd out blood culture. The drug sensitivity test had been done by the method of paper stripdiffusion. The plasmids of bacteria were extracted rapidly by medified Birnboim method and the plasmid analyss was carried out. The plasmids's DNA of 35 epidemic strain was cut off by both restriction enzyme of Hind Ⅲ and EcoR Ⅰ. The outer membrane protein (OMP) was determined by SDS-polyacrylamide gel electrophoresis.Results There are 51 patients with positive blood culture amotile bacterium,of them, pollution; 35 cases (68．6%), septicemia: only 16 cases (31．4%),54．8% (57/104) strains bacteria have drug resistance to more of 12 drugs. 87．3% (165/189) strains bacteria have plasmids. They are cut off as 6 DNA fragments (1．9,2,4,5, 8．5 and 18Kb) by Hind Ⅲ restrietion enzyme. and as 5 DNA fragments (2,2．6,3．2, 6．3 and 22 Kb) by EcoR Ⅰrestrietion enzyme, it is showed that they come from a same clone. The epidemic strain include 10 slips OMP, but non-epidemic strain have 11 slip OMP, increase a 25Kd belt. The amotile bacteria with above-mentioned plasmid spectrum, restriction enzyme spectrum and OMP spectrum are only seen in the air, therapeutic dish and syringe needle.Conclusion The pollution is an important reason of amotile bactorium high positiye rate in new-born.Diagnosing septicemia should depend on bacteria culture, plasmid analysis restriction enzyme analysis of plasmid DNA, oMP determination and combining medical history and clinical manifestation.

Early exercise training after stroke is a therapeutic strategy that has been paid w idespread attention. A series of studies found that early exercise training after stroke can increase expression of neurotrophic factors and maintain the integrity of the blood-brain barrier by inhibiting acute neuroinflammation and apoptosis, and reduce brain damage. This artice reviews the neuroprotective roles of early exercise training after stroke and their possible molecular mechanisms.

OBJECTIVE:To provide reference for the establishment of a system of expired medicine recycling in China. METHODS:The experience of developed countries with respect to the system of expired medicine recycling was summed up,and the existing problems about the recycling of expired medicines in China were considered to make suggestions to the establishment of a system of medicine recycling in China. RESULTS & CONCLUSIONS:Generally,the developed countries such as Britain, France,Germany and America treat expired medicines as hazardous wastes in the Environmental Protection Law,Waste Disposal Law and other laws,and have basically standardized the recycling of expired medicines. The projects of recycling expired medi-cines in those countries are usually funded by their governments or enterprises of manufacturing or selling medicines,where the medicines were recycled at designated places or on a regular basis or by post,executed by pharmacists and pharmacies,and then disposed on the basis of classification or burned and buried at dumps. However,currently there are no laws and regulations and spe-cial funds for medicine recycling in China,leading to people’s lacking of enthusiasm to recycle the expired medicines. It is suggest-ed that China should take such measures as soon as possible,including establishing and improving the laws and regulations of ex-pired medicine recycling,distinguishing the reasonably determination and disposition of different types of expired medicines,accel-erating the reform of the medical insurance payment system,giving full play to the pharmacist’s role of medication guidance,and fully increasing information transparency.

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.

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.