OBJECTIVE To investigate the pathogenic causes of community-acquired pneumonia(CAP) in adult patients in Tongling.METHODS A prospective study was performed on 260 consecutive adult patients with CAP in Tongling city during last three years.Bacteria culture of sputum and serological tests in paired serum samples were detected.RESULTS Of 260 patients with etiological evaluation,128(49.2%) patients had an identifiable etiology,63(24.2%) had positive outcome from sputum cultured,atypical pathogens were detected from 75(28.8%)patients.Pathogens identified in 128 patients were:Mycoplasma pneumoniae(35.4%),Chlamydia pneumoniae(17.7%) and Streptococcus pneumoniae(13.6%).6.5% All patients had mixed infection.The resistance rate of S.pneumoniae to penicillin and erythromycin was 5 and 50%,respectively.CONCLUSIONS Atypical pathogens have important role in CAP,of which M.pneumoniae is the most common pathogen.S.pneumoniae and K.pneumoniae are the commonly encountered bacteria for CAP in Tongling.

Objective To investigate the application effect of stone basket in ureteroscopic holmium laser lithotripsy for treatment of upper ureteral calculi. Methods The clinical data of 96 patients with upper ureteral calculi were retrospectively analyzed, all patients underwent ureteroscopic holmium laser lithotripsy. Forty-eight cases used the stone basket in the operation process (observation group), and 48 cases did not use the stone basket in the operation process (control group). The operation time, length of stay, success rate of lithotripsy, stone residual rate and incidence of postoperative complication were compared between 2 groups. Results The patients of 2 groups successfully completed surgery. There were no statistical differences in operation time, length of stay and incidence of postoperative complication between 2 groups (P>0.05). The success rate of lithotripsy in observation group was significantly higher than that in control group: 97.92% (47/48) vs. 75.00% (36/48), and the stone residual rate was significantly lower than that in control group:4.17%(2/48) vs. 18.75%(9/48), and there were statistical differences (P<0.05). Conclusions The stone basket in ureteroscopic holmium laser lithotripsy for treatment of upper ureteral calculi can thoroughly remove stones. It reduces the incidence of residual stones, does not affect the safety of the operation, and has good clinical value.

Objective To investigate the relationship between the hs-c-reactive protein and the severity of coronary artery disease.Methods All the 67 patients underwent coronary angiography and measured risk factors,the Gensini score was used to determine the results of the coronary angiography.The t test,One-Way ANOVA and multiple linear regression analysis were used to predict hs-CRP.Results Coronary artery disease group hs-CRP levels were significantly higher than those in non-coronary artery disease group(P

Objective To better characterize the clinical and histopathological manifestations, diagnosis and differential diagnosis of epithelioid sarcoma (ES).Methods Clinical data were collected from nine patients with ES diagnosed and treated at the Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College from 2000 to 2014.The clinical and histopathological manifestations, immunophenotype, treatment and prognosis of ES were reviewed retrospectively.Results The median age at onset of ES was 34.5 years in these patients, and the average disease duration was 8.3 years.Lesions began on the extremities in 8 patients, and manifested as nodules,ulceration and lymphedema.Histopathological examination showed that tumor cells mainly consisted of spindle cells and epithelioid cells, some of which were in a palisade arrangement with central necrosis.Cytokeratin, epithelial membrane antigen (EMA) and vimentin were coexpressed by tumor cells.Recurrence was observed at the original site in 6 patients after lesion resection, and pulmonary metastasis occurred in 4 patients.Five patients were followed up and two of them died of pulmonary metastasis.Conclusions Local recurrence is frequent in patients with ES after lesion removal.Lymph node and pulmonary metastases of ES are common, and pulmonary metastasis is usually associated with a poor prognosis.

Objective To investigate clinical and pathological features of primary cutaneous CD30 + anaplastic large cell lymphoma(PC-ALCL). Methods Clinical and pathological data were collected from 7 patients with PC-ALCL and analyzed retrospectively. Results Of the 7 patients, 6 were male and 1 was female, with an average age of 52 years. PC-ALCL was characterized by solitary (n = 3)or multiple (n = 4)erythematous nodules, lumps and/or plaques with (n = 6)or without (n = 1)ulceration. Systemic involvement was observed in none of the 7 patients. Histopathological examination showed diffuse distribution of tumor cells in the dermis, which were large with rich cytoplasm and atypical nuclei. Mitotic figures were seen. An immunohistochemical study of tumor cells showed positive staining for CD30 and cytotoxic protein, but negative staining for CD20, CD56,anaplastic lymphoma kinase(ALK). Epstein-Barr virus-encoded RNA in situ hybridization was negative. Conclusions PC-ALCL is a rare primary cutaneous low-grade malignant T-cell lymphoma, which can be confirmed by clinical manifestations as well as histopathological and immunohistochemical examinations. It usually has good prognosis with rare systemic involvement and metastasis.

Objective: To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods: Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEX-KG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results: The NBEAL1 gene fragment was successfully cloned into pGEX-KG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion: The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.

In order to reaction the quality present situation, problems on the current quality of animal sources of drugs are summed up by using test data analysis, literature search and marketing research. This paper can also help the improvement of the quality management, the revision of the relevant department policy system and the improvement of standards.
Animals ; China ; Medicine, Chinese Traditional ; standards ; Pharmaceutical Preparations ; analysis ; standards ; Quality Control

Objective To construct a eukaryotic expression plasmid vector encoding Cbl-b gene-specific short hairpin RNAs (shRNAs),and to evaluate its interference effect,so as to lay a foundation for further study on the role of Cbl-b in the immunotherapy of malignant melanoma.Methods According to the sequence of Cbl-b cDNA,4 pairs of shRNAs targeting the Cbl-b gene were designed and synthesized,and then inserted into the plasmid PGPU6/GFP/Neo to construct recombinant plasmids.After identification by DNA sequencing,the 4 shRNA expression vectors were cotransfected into 293T cells with the Cbl-b gene eukarytic expresson plasmid,respectively.The knockdown efficiency of these shRNA expression plasmids on Cbl-b expression was evaluated by real-time (RT) fluorescence-based quantitative PCR and Western blot at 48 hours aftert transfection.Results Sequencing analysis revealed that all the 4 pairs of shRNAs were successfully inserted into the eukarytic expression vector PGPU6/GFP/Neo.As RT-PCR and Western blot showed,all the 4 shRNA-expressing vectors could downregulate Cbl-b expession,and the NO.1 shRNA-expressing vector displayed the strongest interference effect(P ＜ 0.05).Conclusions A eukaryotic expression plasmid vector was successfully constructed for Cbl-b gene-specific shRNAs,and the most effective shRNA was selected in this study.

Objective To analyze the incidence of Epstein-Barr virus (EBV) infection in patients after allogeneic hematopoietic stem cell transplantation (HSCT),and investigate its risk factors and prognosis.Methods A total of 402 patients receiving HSCT were retrospectively studied from November 2011 to November 2014 in the 307th Hospital of Chinese People's Liberation Army.The cumulative incidence (CI) of EBV infection and survival rate were analyzed by Kaplan-Meier method,while risk factors were assessed by logistic regression model.Results The one-year CI of EBV viremia and post-transplant lymphoproliferative disease (PTLD) were 42.0％ and 1.5％,respectively.Using antithymocyte globulin (ATG) (P ＜ 0.001,OR =9.92) and acute graft-versus-host disease (GVHD) grade Ⅲ to Ⅳ (P ＜ 0.01,OR =2.42) were related risk factors of EBV viremia.There was a higher CI of EBV viremia in patients with 2 risk factors compared with those without (87.5％ vs 24.6％,P ＜0.001).Patients with EBV viremia had significant shorter three-year overall survival than patients without (58.5％ vs 75.4％,P ＜ 0.001).Conclusions The incidence of EBV infection in patients undergoing HSCT is high,which poses a threat on prognosis.Risk factors of EBV viremia include administration of ATG and severe acute GVHD.

Objective To evaluate in vitro effects of specific small interfering RNA (siRNA)-silencing of the casitas B-lineage lymphoma b (Cbl-b)gene on immunocompetence of primary murine lymphocytes. Methods Spleens were resected from C57BL/6 mice, and splenic lymphocytes were sterily isolated and cultured in vitro. These lymphocytes were divided into 3 groups: silence group transfected with a Cbl-b-specific siRNA using the EntransterTM-R 4000 reagent, negative control group transfected with a negative control siRNA using the EntransterTM-R4000 reagent, blank control group receiving no treatment. After additional culture for 72 hours, ELISA was performed to measure levels of interferon γ(IFN-γ)and tumor necrosis factor α (TNF-α)in culture supernatants of lymphocytes. In addition, the Cbl-b gene-silenced lymphocytes were co-cultured with B16F10 melanoma cells to evaluate their immunocytotoxic effects on melanoma cells. Results Splenic lymphocytes were successfully isolated from C57BL/6 mice and cultured in vitro, and the Cbl-b-specific siRNA was also successfully transfected into the primary murine lymphocytes and effectively down-regulated the expression of Cbl-b gene in them. Compared with the negative control group and blank control group, the silence group showed significantly increased supernatant levels of IFN-γ and TNF-α(all P < 0.05). The immunocytotoxic effect of lymphocytes on melanoma cells was significantly stronger in the silence group than in the negative control group. Conclusion Cbl-b gene silencing can promote secretion of IFN-γ and TNF-α by murine lymphocytes, and enhance their immunocytotoxic effects on B16F10 melanoma cells in vitro.