Objective To investigate whether KAP-1 could induce CSC-self renewal in pancreatic cancer cell lines.Methods KAP-1 expression was examined in 14 cases of pancreatic cancer with immunohistochemistry.KAP-1 shRNA amplified by PCR was inserted into pGC-LV in vector,and then identified by restriction endonuclease digestion and nucleotide sequencing.The lentiviral vector pGC-shRNA-KAP-1 was co-transfected with pHelper 1.0 and pHelper 2.0 packaging plasmids into HEK 293T cells,and the lentivirus was collected and virus titer was measured.The expressions of KAP-1 and vimentin were detected by Western blot and RT-qPCR when human pancreatic cancer cell line Panc-1 was infected by the lentivirus.Sphere forming assay was conducted to assess the capacity of CSC or CSC-like cell self-renewal in this study.Results The KAP-1 expression level in cancerous tissues on immunohistochemistry was significantly higher than in the corresponding normal tissues (P=0.002).After infected by lentivirus,the expressions of KAP-1 and vimentin were knocked down,which could be detected by Wesren blot and RT-qPCR.Compared to Panc-1-GFP (NC) cells,the outcomes suggested that knocking down the expression of KAP-1 could decrease the formation of pancreatospheres,which further suggested the capacity of CSC-self-renewal in primary and secondary pancreatospheres of Panc-1-shRNA-KAP-1 and NC cells.Conclusions KAP-1 expression in pancreatic cancer tissues has been identified.The lentiviral vector for shRNAs targeting KAP-1 was constructed successfully.The formation of pancreatospheres decreased by knocking down the KAP 1 gene.KAP-1 is involved in the regulation of CSC phenotype.The mechanism is probably related to the upregulated expression of the EMT marker vimentin.

Objective To investigate the effects of KAP-1 in promoting the epithelial-mesenchymal transition (EMT) of pancreatic cancer cell line Capan-2.Methods The lentiviral vector of LV-plenti-GFP-KAP-1 was constructed.Capan-2 cells were divided into the experimental group (cells transfected by lentiviral vector of LVplenti-GFP-KAP-1),negative control group (cells transfected by empty vector) and blank control group 1 (cells cultured in 1640 medium plus 10％ fetal calf serum).Capan-2 cells in the experimental group were subdivided into the miR-100-5p inhibitor transfection group (cells transfected with miR-100-5p inhibitor),empty vector control group (cells transfected with microRNAs inhibitor),blank control group 2 (cells cultured in 1640 medium plus 10％ fetal calf serum).The lentivirus was identified by double endonuclease restriction and sequencing,and the virus titer was detected.The morphological changes of the cells were observed after transfecting lentiviral vector of LV-plenti-GFP-KAP-1 to the Capan-2 cells.The expressions of KAP-1,genes of EMT proteins and mRNA of miR-100-5p were detected by real-time quantitative polymerase chain reaction.The protein expressions of KAP-1,EMT proteins in all the groups were detected by Western blot.The measurement data were presented by mean ± standard deviation,and were analyzed using the analysis of variance.Results The lentiviral vector of LV-plentiGFP-KAP-1 was successfully constructed,and the virus titer was 2 × 108 TU/ml.Compared with the control group,the mesenchymal transition of the Capan-2 cells was detected in the experimental group after transfecting the Capan-2 cells with lentiviral vector of LV-plenti-GFP-KAP-1 for 48 hours.The relative mRNA expressions of KAP-1,N-cadherin,vimentin,E-cadherin,miR-100-5p were 1.77 ± 0.83,2.62 ± 0.71,2.50 ± 0.21,7.20 ± 1.17 and 1.81 ±0.40 in the experimental group,5.03 ±0.29,5.07 ±1.53,3.83 ±0.57,7.83 ±0.78,7.01 ± 0.96 in the negative control group,5.13 ± 1.14,5.81 ± 1.49,4.92 ± 0.90,3.07 ± 0.36,6.87 ± 0.35 in the blank control group 1,with significant difference among the 3 groups (F =5.99,7.62,7.88,6.62,4.64,P ＜0.05).The relative mRNA expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 1.56 ± 0.42,4.89 ± 0.61,5.20 ± 0.38,with significant difference among the 3 groups (F =5.14,P ＜ 0.05).The relative mRNA expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.10 ± 1.37,3.44 ± 0.94,3.08 ±1.16,with no significant difference among the 3 groups (F =0.49,P ＞ 0.05).The results of western blot showed that the relative protein expressions of KAP-1,N-cadherin,vimentin,E-cadherin were 2.77 ± 1.99,1.31 ±0.38,4.25 ± 0.63,0.62 ± 0.06 in the experimental group,0.83 ± 0.46,0.41 ± 0.37,1.03 ± 0.33,1.17 ± 0.45 in the negative control group,0.71 ± 0.26,0.08 ± 0.04,1.37 ± 0.92,3.04 ± 0.65 in the blank control group 1,with significant difference among the 3 groups (F =5.54,4.68,3.19,8.18,P ＜ 0.05).The relative protein expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 2.27 ±0.71,0.56 ±0.43,0.61 ±0.39,with significant difference among the 3 groups (F =4.81,P ＜0.05).The relative protein expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.19 ± 0.55,3.93 ± 0.06,3.61 ± 0.73,with no significant difference among the 3 groups (F =0.04,P ＞ 0.05).Conclusion KAP-1 promotes the EMT of Capan-2 cells by specifically down-regulating the miR-100-5p expression.

Objective To study the clinical value of HRCT in staging of lung interstitial disease(LID) in connective tissue disease(CTD).Methods 222 patients with CTD confirmed clinically underwent HRCT scan.The staging of LID according to the HRCT features of LID were done and the therapeutic effect was compared between each stage.Results In 222 cases ,64 cases were negative on HRCT as stage 0,158 cases had disseminated LID in different degree,including stage Ⅰ in 107,stage Ⅱ in 36 and stage Ⅲ in 15.There were significant statistically between the therapeutic effect and staging of LID.Conclusion The staging of CTD with LID by HRCT is helpful for judging the extent of LID and clinical treatment.

Object To observe the effect of Guizhi Fuling Capsule (GFC) on prostatic hyperplasia in rats induced by testosterone propionate. Methods Seven days after the male rats were castrated, testosterone propionate (3 mg/kg) was sc injected in rats. Meanwhile, the rats were administered GFC (2.16, 1.08, 0.54 g/kg) once a day, and the treatment continued for 30 d. An hour after the final administration, the rats were put to death, and the weight and volume of prostates were measured. The construction of prostate cells was also observed under light microscope. Results In the GFC group, the weight and volume of prostate was significantly decreased compared with that in the model group (P

Objective This study was to analyze the early and long-term effect of completion pneumonectomy.Methods Retrospective analysis was made on the patients who underwent completion pneummonectomy in Shanghai Chest Hospital.Results There were totally 56 cases patients underwent completion pneumonectomy during January 2003 to July 2013.Among them,45 patients received CCP,and other 11 patients received RCP.CCP refers to the complete removal of lung tissue remaining after an initial ipsilateral partial pulmonary resection.RCP refers to the complete removal of residual lung due to the severe complications after pneumonectomy.The mortality and morbidity rate of CCP were 4.4％ and 33.3％ respectively.In the case of CCP,the incidence of benign lesions is significantly higher than the incidence of malignant tumor(80.0％ vs 27.5％,P =0.04).The mortality and morbidity rate of RCP were 27.3％ and 90.9％ respectively.In the case of RCP,higher postoperative mortality often occurs in aged patients (P =0.046) and patients with preoperatie mechanical ventilation (P =0.03).Overall five-year survival rate for patients with benign lesions was 80％,and for malignant lung cancer patients,the number was 30％.Survival time differs according to the TNM staging(a median of 60.0 months,35.0 months,10.0 months,stage Ⅰ,stage Ⅱ,stage Ⅲ,P ＜0.01),and survival rate was higher when the time interval(between the initial pulmonary resection and the completion pneumonectomy) ＞ 2 years(a median of 60.0 months,18.0 months,P ＜ 0.01).Conclusion Completion pneumonectomy is a high-risk surgery,especially RCP.Advanced age and preoperative mechanical ventilation are associated with higher postoperative mortality rate for RCP.As for CCP,higher postoperative risk exists in patients with benign lesions,but the survival rate is also higher.In patients with malignant lung tumor,survival rate is higher when the time interval (between the initial pulmonary resection and the completion pneumonectomy) ＞2 year.

Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.

Acute leukemia marks the main type of non-solid cancer in children.Studies have proved that long non-coding RNA plays important roles in the pathogenesis of hematological malignancies and can be applied as a biomarker for diagnosis, prognosis and efficacy monitoring of leukemia patients.This article briefly reviews the roles of long non-coding RNA in the tumorigenesis and progression of childhood acute leukemia.

Objective To investigate the role of the group IB secretory phospholipase A2 (sPLA2-IB) and M-type phospholipase A2 receptor (PLA2R) in human podocyte injury and its possible signal transduction pathway.Methods Differentiated human podocytes were exposed to PBS or different sPLA2-IB concentration conditions (10-9,10-7,10-5 mol/L) for 2 hours.The wound healing assay was used to measure cell migration rate;Apoptosis in cultured human podocytes was assessed by Hoechst 33342 staining and flow cytometry;Western blot was used to analyze the protein expression of p-cPLA2α,p-p38,and p53.Then control siRNA or PLA2R siRNA were transfected to podocytes.Podocytes were divided into normal control group,negative control siRNA group and PLA2R siRNA group.Twenty four hous later,the cells were stimulated by 105 mol/L sPLA2-IB for 2 hours.The protein expression of p-cPLA2α,p-p38,and p53 were detected by Western blot.Results Compared to PBS control group,the migration ability of podocytes decreased when stimulated with sPLA2-IB (10-7mol/L-10-5 mol/L),and the apoptosis of podocytes increased in a concentration-dependent manner,the protein level of p-cPLA2α,p-p38 and p53 protein increased too.After the knockdown of PLA2R by PLA2R siRNA transfection,stimulated the podocytes with the same dosage of sPLA2-IB,the protein expression of p-cPLA2α,p-p38 and p53 all decreased.Conclusion sPLA2-IB stimulation can increase human podocyte apoptosis and decrease its migration ability.The possible mechanism might be through p38-cPLA2α-p53 pathway.

Objective:To survey the working status of outpatient nurses in general practice departments of general hospitals.Methods:From March to April 2021, personal in-depth interviews were conducted with outpatient nurses from general practice residency training bases in Beijing. The thematic framework analysis method was used to analyze the interview data and refine the themes.Results:Fourteen nurses working in general practice outpatient clinics were interviewed in this study. Four themes were extracted from the interview data: (1) inadequate staffing of full-time outpatient nurses in general practice departments of comprehensive hospitals; (2) unclear job functions for outpatient nurses in general practice departments; (3) no standardized patient health education in the department; (4) no relevant training received systematically.Conclusion:General practice departments in general hospitals should setup full-time general practice nurse positions, clarify the job responsibilities, strengthen the relevant training and enhance core competency for nurses working in general practice department.

Since the performance appraisal of national tertiary public hospitals was carried out, higher requirements have been put forward for the operation and management of hospitals. Under the premise of ensuring the quality of medical service and medical safety, how to save hospital operating costs and improve the efficiency is an urgent problem for hospital managers. Supported by information upgrading, a tertiary hospital in Guangzhou reformed the treatment process and carried out pre-hospitalization in surgical departments. Data showed that pre-hospitalization can significantly shorten the length of stay, reduce hospitalization costs, and improve the operation efficiency of the hospital.