Objective The aim of this research is to clone cDNA of human prostate-specific membrane antigen(PSMA),construct pcDNA3-PSMA eukaryotic expression vector, and then identify the expression of PSMA protein. We hope this experiment will give a good basis on the research of the prostate therapy based on DC vaccine. Methods Total RNA was extracted from prostate cancer tissues. A pair of primers were designed according to M99487 in Genebank and obtain the PSMA cDNA fragment by reverse transcription-polymerase chain reaction. The PCR products were then cloned into empty plasmid pcDNA3.Further sequence analysis was carried out and the recombinant plasmids PcDNA3.0-PSMA were transfected into PC-3 cells. In G418 selective medium(400μg/ml),two positive clones expressing PSMA mRNA, named D4 and B3 respectively, were identified by RT-PCR. Using rabbit anti-human PSMA monoclonal antibody, we identified the location of expressed PSMA protein by indirect immunofluorescence analysis. To verified the molecular weight of expressed PSMA protein, western blotting analysis of total cell extracts was carried out using chemiluminescence western blotting kit. Results Sequence analysis confirmed the homogeneity between cloned PSMA cDNA and M99487 was 99.7%.After transfection. two cell clones expressing PSMA mRNA were identified by RT-PCR.The indirect immunofluorescence assay confirmed that expressed PSMA protein was located on membrane and had good immunoreactivity. Western blotting analysis showed the molecular weight of expressed PSMA was 100kD.as was PSMA protein from LNCaP cells. Conclusion We have obtained the PSMA cDNA from prostate cancer tissues. The PcDNA3.0-PSMA eukaryotic expression vector was constructed and the target PSMA protein which had immunoreactivity expressed on cell membrane. The molecular weight of expressed PSMA was 100kD,the right molecular weight as we expected. Our research layed foundation for treatment of prostate neoplasms by DC vaccines transfect with PSMA.

Objective To evaluate the pathogenic bacteria and drug resistance variation of bone and joint pyogenic infection, and to provide reference for clinical antibiotic therapy. Methods A total of 172 cases with bone and joint pyogenic infection from Jan 2005 to Oct 2007 in the Department of Or- thopedics were retrospectively analyzed with the results of bacterial cultures and antibiocties sensitivity tests of blood, purulence or hydrothorax samples. Results The detection rate of Staphylococcus epi- dermidis, Staphylococcus aureus, Escherhia coil, Acinetobacter baumannii and Enterococcus faecalis accounted for 24.4%, 19.3%, 18. 5 %, 8. 1% and 7.4% respecterly. Conclusion Staphylococci are the major pathogenic bacteria leading to bone and joint pyogenic infection, followed by Escherhia coil, Pseudomonas aeruginosa and fungi. It is suggested that sensitive antibiotics be selected according to drug susceptibility test results. To staphylococcal infection cases, combined application,of antibiotics may improve the therapeutic effect, prevent occurrence of drug resistance, and reduce drug adverse re- action. Early and standard antibiotic therapy and timely local drainage of infection focus contribute to reduce antibiotic resistance and incidence of chronic diseases.

Objective To discuss the CT findings of the ovarian endometriosis cyst.Methods CT findings of 15 cases ovarian endometriosis cyst confirmed by patology were retrospectively analyzed.Results There were 20 cysts in 15 cases.Their CT features were as follow:(1)CT values varied from watery to hemoid.(2)They had a irruglar outer fringe and a regular inner fringe without noduls.(3)Septations were seen in the cysts.(4)The characteristic feature was that daughter cysts lied outside of mother cysts.(5)The wall of cysts could be enhanced regularly.Conclusion CT plays an important role in the diagnosis of this disease.

Comparative genomic hybridization is a techniqu e combining fluorescence in situ hybridization with subtractive hybridization in molecular-cytogenetics. The technical basic priciple, methods, quality control , advantages,disadvantages, applications and advances in the study of colorectal carcinoma are reviewed.

Objective To investigate the effects of myopia on first-order kernels six ring responses of multifocal electroretinogram (mfERG). Method We continuously selected 80 subjects aged from 13 to 20 year-old and divided four groups into normal, mild myopia, moderate myopia and high myopia. Each group had 20 subjects that were examined with mfERG of 103 stimuli locations. The amplitude and latency of N1, P1, N2 waveforms of six-ring first-kernel responses in mfERG were analyzed. Result With increasing of myopia degree, the response density of N1, P1, N2 waveforms of six-ring first-kernel responses in mfERG was decreased gradually (all P0.05). Conclusion Myopia can decrease the response density of first-order kernels six ring responses of mfERG, which is very important to determine the abnormality of mfERG in the clinic.

In order to service clinical departments, reduce the cost and improve the efficiency of the storage, on the basis of information system, the overall planning of the requirement of frameworks and internal infrastructures of the hospital logistics center are proposed. The requirements of supplier admittance, the quality and the activities of the storage, the storage ratio of materials and the storage expense are discussed. It is pointed out that the integration management of logistics center, access control items, the reasonable management of storage materials, analyzing, evaluating and adjusting of various management indicators are effective methods of product quality assurance, reducing storage losses and monitoring costs.

Objective To assess the effect of arterial embolization in treating hyperthroidism by colour ultrasonography.Methods Forty two cases of hyperthyroidism were treated with thyroid arteries embolization. A few days before and 1, 3, 6 months after embolization, the echograms of thyroids were observed including the volumes of thyroids and the internal diameters of thyroid arteries were measured with colour ultrasonography respectively. The Vs, Vd, Vm, PI, RI were measured with the Doppler and the quantities of blood flow were calculated. The relationships of changes for all these parameters and T 3, T 4 and TSH were analysed. Results Before artery embolization all thyroids were enlarged with diffusely homogenous or heterogenous low echos and nodules in some patients accompanied by widenings of the thyroid arteries and their branches full of blood supply. The volume of thyroids decreased after artery embolization. 1 3 months later the echo of thyroids enhanced and got coarse with decrease of the thyroid vasculature and narrowing of vessel calibers except a few star or spot like blood streams were sometimes seen within the thyroids and no blood flow signals found in some cases. 3 6 months after embolization, the echo of thyroids decreased gradually or unevenly distribnted. All the parameters of blood flow before and after the treatment showed statistically significant differences( P

OBJECTIVE: To investigate the influence of signal transducer and activator of transcription 3 (Stat3) and hypoxia-inducible factor-lα (HIF-1α) on the resistance effect of laryngeal squamous cell carcinoma to radiation therapy and chemotherapy under the hypoxia circumstances. METHOD: Western blot was used to test the expression of p-Stat3 and HIF-1α in the Hep-2 cells under the hypoxia conditions. MTT assay was used to test the proliferation of Hep-2 cells after radiation therapy and chemotherapy; the Hep-2 cells were suppressed expression of Stat3 and/or HIF-1α. RESULT: (1) AG490 induced significant proliferation inhibition on Hep-2 cells and Hep-2HIF-1α-/- cells in vitro underthe hypoxia environments (P < 0.05); (2) Suppressing expression of Stat3 reduced the expression of HIF-1α protein (P < 0.05); (3) Combined inhibition of Stat3 and HIF-1α enhanced radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypoxia. CONCLUSION Combined inhibition of Stat3 and HIF-1α can further enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypixia compare than inhibiting Stat3 or HIF-1α alone. Effectively blocking of HIF-1α pathway and suppressing the expression of Stat3, would be an effective method to enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells, which provides a new thought to reduce the resistance to treatment.
Blotting, Western ; Carcinoma, Squamous Cell ; Cell Hypoxia ; Cell Line, Tumor ; Humans ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit ; Laryngeal Neoplasms ; STAT3 Transcription Factor

Objective: To evaluate 2D CDUS ultrasound in estimating lesions of carotid and femoral arteries in diabetes mellitus. Methods: Forty patients were divided into 3 groups by age. We detected their carotid and femoral arteries using two dimensional and PW Doppler ultrasonography. The lesions degree were assessed according to U B 6 classification. Results: Higher incidence and more serious degree of arteries lesions occurred in diabetes mellitus than those in control group ( P 40 years old than in group ≤40 years old in diabetes mellitus ( P

OBJECTIVE:To discuss the drug resistance situation and influential factors of sputum culture positive tuberculosis patients in a specialized subject hospital. METHODS:The information of sputum culture positive tuberculosis inpatients were col-lected from Heze infectious disease hospital during Jan. 1st 2014 to Apr. 30th 2015;the results of drug sensitive test were analyzed retrospectively by using absolute concentration method. RESULTS:Among 259 patients,137 cases were drug-resistant(52.90%), among including 69(26.64%) mono-drug resistant cases,52(20.08%) poly-drug resistant cases,15(5.79%)multi-drug resistant cases and 1(0.39%)extensively drug resistant case;streptomycin (31.66%) and amikacin (15.44%) were drugs with the highest drug-resistant rate;there was no statistical significance in resistant rate between different genders and ages(P>0.05),the rate of drug resistance in first-treatment patients was 47.21%,and that of the re-treatment patients was 70.97%,the difference was statisti-cal significant(P<0.05). In addition to ethambutol and kanamycin,other all had mono-drug resistance,there were 27 combinations of poly-drug resistance and 9 combinations of multi-drug resistance. Logistic regression analysis showed that treatment history was a risk factor for drug-resistance,the risk of re-treatment patient was 2.734 times of first-treatment patients. CONCLUSIONS:Sputum culture positive tuberculosis inpatients patients have high drug resistance and complex spectrum in the hospital;we should pay great attention to the risk of drug resistance in senile patients and re-treatment patients,and rational use of streptomycin,amikacin,isoni-azid and capreomycin.