Objective: To investigate the effect and mechanism of phosphoprotein phosphatase 5 catalytic(PPP5C) on the migration , invasion and tumor stemness of human lung adenocarcinoma H1299 cells. Methods: The PPP5C⁃pcDNA3. 1 overexpression vector was constructed. PPP5C⁃pcDNA3. 1 and pcDNA3. 1 were transfected into H1299 cells , and H1299 stable cell lines were screened with G418. The mRNA and protein expression levels of PPP5C were identified by qRT⁃PCR and Western blot. The proliferation activity of H1299 cells was detected by drawing cell growth curve and cell clonal formation assay. The wound⁃healing assay and transwell assay were used to test the migration and invasion abilities of H1299 cells , respectively. The stemness of H1299 cells was evaluated by sphere formation assay. Results: The PPP5C⁃pcDNA3. 1 eukaryotic expression vector was successfully constructed and the expression levels of PPP5C significantly increased after transfection into H1299 cells. After overexpression of PPP5C in H1299 cells , the cell growth curve and clonal formation assay displayed that the proliferation ability was not affected , the migration and invasion of cells were significantly enhanced through wound⁃healing assay and transwell assay , accompanied by an increase in the expression of MMP9 , stem cell spheroid assay showed a significant increase in stemness of cells , accompanied by increased expression of SOX2. Conclusion The proliferation ability of cells is not affected , the migration and invasion and the stemness of cells are enhanced by regulating MMP9 and SOX2 respectively , after overexpression of PPP5C in human lung adenocarcinoma H1299 cells.

Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

Objective:Based on the investigation of students after the "periodic recall" examination, to discuss the role and existing problems of this kind of examination in the joint training of standardized training for residents.Methods:In March and October, 2017, questionnaire survey was conducted on the students who participated in the "periodic recall" clinical examination, and the arrangement and implementation of the assessment were evaluated by the students in terms of satisfaction, and the ability status before and after the clinical examination was assessed in person on a scale of 1-10. At the same time, the results of the two times of examinations were analyzed, and the average scores of all skills and the overall average scores were calculated to compare the differences. SPSS 16.0 was used for t test and variance analysis. Results:A total of 112 questionnaires were distributed and 99 were returned. Students' overall satisfaction with the examination arrangement was high, with the satisfaction of all items above 70%. After the examination, the students' self-assessment scores of all abilities were improved, and there were significant differences before and after the examination. There was a significant difference in the average improvement of self-assessment ability among the seven skills ( F=4.19, P=0.006). The results analysis showed that the average score of simple operation was higher than the overall average score, with significant differences. Conclusion:"Periodic recall" examination can meet students' psychological and learning needs in the tumor hospital training, with high satisfaction, which has obvious effect on improving students' simple operation skills, but weak improvement of complex comprehensive skills.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.

The BCL6 (B-Cell Lymphoma 6) gene is a proto-oncogene that is often expressed in diffuse large B-cell lymphomas (DLBCLs). BCL6 loss of function can kill DLBCL cells, demonstrating that BCL6 is necessary for the survival of DLBCL cells and could be a therapeutic target. In this study, we found that BCL6 protein levels were consistently upregulated in DLBCL tissues, whereas its mRNA levels varied randomly in tissues, suggesting that a post-transcriptional mechanism was involved in BCL6 regulation. We used bioinformatics analysis to search for miRNAs, which potentially target BCL6, and identified specific targeting sites for miR-10a in the 3'-untranslated region (3'-UTR) of BCL6. We further identified an inverse correlation between miR-10a levels and BCL6 protein levels, but not mRNA levels, in DLBCL tumor tissue samples. By overexpressing or knocking down miR-10a in DLBCL cells, we experimentally validated that miR-10a directly recognizes the 3'-UTR of the BCL6 transcript and regulated BCL6 expression. Furthermore, we demonstrated that negatively regulating BCL6 by miR-10a suppressed the proliferation and promoted apoptosis of DLBCL cells.
3' Untranslated Regions ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; therapy ; MicroRNAs ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; biosynthesis ; genetics

Neutrophils play an essential role in the innate immune response to infection. Neutrophils migrate from the vasculature into the tissue in response to infection. Recently, a neutrophil cell surface receptor, CD177, was shown to help mediate neutrophil migration across the endothelium through interactions with PECAM1. We examined a publicly available gene array dataset of CD177 expression from human neutrophils following pulmonary endotoxin instillation. Among all 22,214 genes examined, CD177 mRNA was the most upregulated following endotoxin exposure. The high level of CD177 expression is also maintained in airspace neutrophils, suggesting a potential involvement of CD177 in neutrophil infiltration under infectious diseases. To determine the role of CD177 in neutrophils in vivo, we constructed a CD177-genetic knockout mouse model. The mice with homozygous deletion of CD177 have no discernible phenotype and no significant change in immune cells, other than decreased neutrophil counts in peripheral blood. We examined the role of CD177 in neutrophil accumulation using a skin infection model with Staphylococcus aureus. CD177 deletion reduced neutrophil counts in inflammatory skin caused by S. aureus. Mechanistically we found that CD177 deletion in mouse neutrophils has no significant impact in CXCL1/KC- or fMLP-induced migration, but led to significant cell death. Herein we established a novel genetic mouse model to study the role of CD177 and found that CD177 plays an important role in neutrophils.
Animals ; Disease Models, Animal ; GPI-Linked Proteins ; genetics ; Gene Expression Regulation ; Genetic Therapy ; Humans ; Immunity, Innate ; genetics ; Inflammation ; genetics ; microbiology ; pathology ; Isoantigens ; genetics ; Mice ; Mice, Knockout ; Neutrophils ; metabolism ; pathology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Cell Surface ; genetics ; Staphylococcus aureus ; pathogenicity ; Transcriptional Activation

Objective To evaluate the efficacy and safety of oxycodone hydrochloride injection for postoperative analgesia in patients undergoing the operation under general anesthesia in a prospective,randomized,blind,multicenter,positive-controlled,clinical trial.Methods Two hundred and forty ASA Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,weighing 40-95 kg,scheduled for elective abdominal operation or orthopedic surgeries under general anesthesia,were randomly divided into 2 groups (n =120 each):morphine sulfate injection group (group M) and oxycodone hydrochloride injection group (group O).Morphine or oxycodone 1 mg was injected intravenously when the patients complained of pain after tracheal extubation or removal of the laryngeal mask,and administration was repeated if necessary until VAS≤40 mm.Then patient-controlled intravenous analgesia (PCIA) (100 ml,0.5 mg/ml) with morphine or oxycodone was used for postoperative analgesia (lasting for 48 h).The PCIA pump was set up with a 1 ml bolus dose,a 5 min lockout interval and background infusion at a rate of 0.5 mg/h.Pain at rest and during movement was assessed using VAS score at 3,24 and 48 h after administration,and non-inferiority test was performed.Total morphine or oxycodone consumption,requirement for rescue analgesic,the number of unsuccessfully delivered dose,the number of attempts,and the level of patient' s satisfaction were recorded within 48 h after operation.The adverse events were recorded and laboratory examinations (blood and urine routine test,blood biochemical examination) were performed within 72 h after administration.Results There was no significant difference in the VAS scores at rest and during movement at different time points,requirement for rescue analgesic,the number of unsuccessfully delivered doses and attempts,level of patient' s satisfaction,total morphine or oxycodone consumption,and adverse events between the two groups (P ＞ 0.05).No serious adverse event occurred in the two groups.The most common adverse event was nausea,followed by vomiting.There was no significant difference in the incidences and degree of nausea and vomiting between the two groups (P ＞ 0.05).The incidences of nausea and vomiting in patients underwent orthopedic surgeries were significantly lower in group O than in group M (P ＜ 0.05).The other adverse events were fewer and abnormal laboratory examinations were rare in the two groups.95％ confidence interval of the difference between the mean VAS scores at rest and during movement at each time point was within 15 mm (boundary values of non-inferiority testing) in the two groups.Conclusion PCIA with oxycodone hydrochloride injection is safe and effective in reducing pain after moderate or major operation,and the analgesic efficacy is similar to that of morphine sulfate injection,however,the development of nausea and vomiting is reduced when PCIA with oxycodone hydrochloride injection is used for orthopedic surgeries as compared with that when morphine sulfate injection is used and the ratio between the analgesic efficacy of the two drugs is close to 1∶1.

Aim To investigate the protective effects of high concentration fentanyl on the brain slice injury induced by oxygen glucose deprivation(OGD).Methods Rat brain slices were made and randomly assigned to four groups:control(n=10),OGD(n=10),fentanyl 50 ?g?L~(-1)(F_(50),n=10) and fentanyl 500 ?g?L~(-1)(F_(500),n=10).Changes of the neuron injury and apoptosis were observed with TTC staining,LDH releases,TUNEL staining,immunohistochemistry and electromicroscope.In addition,changes of intracellular calcium were measured with confocal laser-scanning microscopy.Results F_(50) and F_(500) attenuated the decrease of TTC staining and the increase of LDH release induced by OGD in brain slices.Neuronal apoptosis and changes of neuronal ultrastructures were attenuated by F_(50) and F_(500).Bcl-2 and Bax protein expressions were increased after OGD.Bax protein expression was decreased by F_(50) and F_(500),while Bcl-2 protein expression was increased by F_(50)and F_(500).Intracellular calcium concentration was increased by OGD and then it was lowered by F_(50) and F_(500).The protective effects of F_(50) were more obvious than that of F_(500).Conclusions High concentrations of fentanyl have neuron protective effects against OGD injury in rat brain slices,and fentanyl 50 ?g?L~(-1) has more obvious protective effects than fentanyl 500 ?g?L~(-1).

Objective To evaluate the accuracy of a new generation bioimpedance cardiac output monitor as compared with thermodilution and end expiratory CO 2 techniques Methods Sixteen patients (14 males, 2 females) aged (58?12) years undergoing elective coronary artery bypass (CAB) surgery were studied Cardiac output was measured after CAB surgery in ICU using a new generation bioimpedance cardiac output monitor(Rheo Bioimpedance Hemodynamic monitor) and conventional thermodilution technique in 16 patients and end expiratory CO 2 technique in 10 patients Cardiac output was also measured with Rheo Bioimpedance Hemodynamic monitor in 200 healthy volunteers consisting of equal numbers of male and female The volunteers of each sex were further divided into 5 equal groups of 20 each: (1)20 29 years, (2)30 39 years, (3)40 49 years, (4)50 59 years and (5)60 70 years Results Cardiac outputs measured by bioimpedance and thermodilution techniques were well correlated (r=0 83) and there was also a good correlation between cardiac outputs measured by bioimpedance and end expiratory CO 2 techniques(r=0 88) Cardiac out measured with Rheo Bioimpedance Hemodynamic monitor in 200 healthy volunteers showed that there was a significant difference in cardiac output between 60 70 years age group and the other age groups in both male and female Conclusions Cardiac output and other hemodynamic parameters measured with the new generation bioimpedance hemodynamic monitor are accurate and can be used for preoperative cardiovascular function evaluation and postoperative monitoring of dynamic changes in circulatory function