Objective To study the effect of Mycobacterium tuberculosis (Mtb) Pro-Pro-Glu-59 (PPE59) protein on the biological function of Mycobacterium smegmatis (Ms) and the regulation of host cell immune response. Methods PPE59 gene fragment was obtained by PCR amplification, cloned into pALACE, constructed into recombinant pALACE-PPE59 vector, and electro-transformed into Ms. Western blot was applied to analyse PPE59 expression and subcellular localization. The survival of Ms_Vec and Ms_PPE59 under low acid (pH=3 and pH=5) conditions and active surface pressure sodium dodecyl sulfate (SDS) conditions and their intracellular survival in macrophages were analyzed. ELISA was used to detect the cytokine (IL-1β, IL-6, IL-12, TNF-α and IL-10) expression levels of Ms_Vec and Ms_PPE59 infected macrophages. Results PPE59 protein localized to the cell wall of Ms can enhance the acid-resistance and anti-SDS effect of Ms, which is conducive to the survival of Ms in macrophages. PPE59 significantly decreased the secretion levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-12 and TNF-α), and promoted the secretion levels of anti-inflammatory cytokine (IL-10). Conclusion PPE59 enhances the survival ability of Ms under low acid and SDS pressure and promotes its intracellular survival by regulating the cytokine secretion levels.
Mycobacterium smegmatis/metabolism* ; Macrophages/metabolism* ; Cytokines/metabolism* ; Mycobacterium tuberculosis/metabolism* ; Bacterial Proteins/metabolism* ; Animals ; Mice ; Antigens, Bacterial/metabolism*

Humans ; Intra-Abdominal Fat/diagnostic imaging* ; Coronary Artery Disease ; Body Mass Index

Objective:To summarize the best evidence for follow-up patients after heart transplantation so as to provide a reference for clinical practice.Methods:According to the "6S" evidence model, we systematically searched the National Guideline Clearinghouse, National Institute for Health and Clinical Excellence, Scottish Intercollegiate Guidelines Network, Clinical Practice Guideline of Canadian Medical Association, Australian Government National Health and Medical Research Council, Guidelines International Network and Medlive and other guide networks, and PubMed, CINAHL, Embase, UpToDate, British Medical Journal (BMJ) best practice, Joanna Briggs Institute (JBI) Evidence-based Practice Database, Cochrane Library, China National Knowledge Infrastructure (CNKI) , Wanfang Database, Chinese Biology Medicine Literature, VIP and other databases for the evidence on the follow-up of patients after heart transplantation. The evidence included systematic reviews, guidelines, and expert consensus. The search time limit was from the establishment of the database to October 2020. Two researchers independently evaluated the quality of literature, and extracted and summarized the evidence that met the quality standards.Results:A total of 10 articles were included, including 2 guidelines, 7 systematic reviews, and 1 expert consensus. A total of 26 pieces of evidence were collected from 8 topics including follow-up team composition and requirements, follow-up frequency, follow-up content, condition monitoring, evaluation and improvement of treatment compliance, evaluation and improvement of psychological conditions, exercise and rehabilitation, and prevention and management of complications.Conclusions:This study summarizes the best evidence for follow-up patients after heart transplantation. Medical and nursing staff can form effective and feasible nursing strategies according to the patient's wishes and specific clinical conditions, combined with professional judgment, so as to implement scientific follow-up management of patients.

Objective:To establish osimertinib-resistant non-small cell lung cancer (NSCLC) cell line and explore its drug resistance mechanism.Methods:The human NSCLC cell line H1975 was used as the research object, and low-concentration osimertinib was used to continuously select secondary drug-resistant cell lines. Osimertinib drug sensitivity of cells was detected by MTS method. Cell proliferation was detected by live cell workstations. Flow cytometry was used to detect cell cycle and apoptosis. Protein mass spectrometry was used to construct differentially expressed protein profiles between parental and drug-resistant cells and some resistance-related proteins were validated by real time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot.Results:Secondary drug-resistant H1975/OSI cell line were successfully established. Compared with the parental cells, the resistance index of H1975/OSI cells increased by 27.25 times ( P<0.01), the cell proliferation ability decreased but the apoptosis resistance increased ( P=0.01), and no new drug-resistance related gene mutation in H1975/OSI cells. Meanwhile, the differential protein expression profiles of H1975 and H1975/OSI cells were built, and 307 upregulated proteins and 295 down-regulated proteins were found in resistant cells. When fibroblast specific protein-1 (FSP1) gene with expression up-regulation was diturbed in H1975/OSI cells, the cell IC50 value of osimertinib decreased 3.51 times ( P=0.02) , and when FSP1 was overexpressed in the H1975 cells, the IC50 value of osimertinib increased by 3.75 times ( P<0.01). Conclusions:We successfully established human NSCLC osimitinib-resistant cell line H1975/OSI. Protein differential expression profiles between H1975 and H1975/OSI was constructed successfully. It was found that FSP1 was involved in mediating the resistance of H1975/OSI to osimertinib.

Objective To investigate the effect and mechanism of miR-23b on the proliferation and migration of triple negative breast cancer (TNBC) cells.Methods Real time polymerase chain reaction (PCR) was used to detect the expression of miR-23b in triple negative breast cancer tissues.MDA-MB-231/miR-23b,BT549/miR-23b cell lines are constructed.Proliferation assay,scaling healing experiment and Transwell migration assay were used to detect the effect of miR-23b on the proliferation and migration of triple negative breast cancer cells.Dual-luciferase reporter gene assay was employed to examine the interactions between miR-23b and forkhead box C2 (FOXC2).Real time PCR and Western blot were performed to detect the effect of miR-23b on the expression of FOXC2.Results The expression level of miR-23b in triple negative breast cancer tissues was significantly less than that in adjacent normal tissues.miR-23b could reduced the proliferation and migration of triple negative breast cancer cells.Dual-luciferase assay confirmed that miR-23b could regulate the expression and activity of FOXC2.The expression of FOXC2 in mRNA and protein level was inhibited by miR-23b.Conclusions miR-23b can inhibit the expression of FOXC2 and affect the proliferation and migration of triple negative breast cancer cells.

Objective To investigate the related factors of serum carnitine deficiency in critical ill patients, and the influence of its deficiency on the length of hospital stay. Methods A prospective study was conducted. Critical ill patients with acute physiology and chronic health evaluationⅡ(APACHEⅡ)score>12 admitted to Department of Critical Care Medicine of the First Affiliated Hospital of Sun Yat-sen University from March 2013 to September 2013 were enrolled. Serum carnitine concentration and indexes of organ function were determined,and the tolerance of enteral nutrition within 5 days,the length of hospital stay,the length of intensive care unit(ICU)stay,and the hospital mortality were recorded. The relationship between serum carnitine and indexes mentioned above was analyzed. Results Thirty critically ill patients were enrolled. Serum carnitine concentration was very low in all critically ill patients,i.e. (8.92±5.05)μmol/L(normal reference value at 43.5 μmol/L)at hospital admission. Serum carnitine concentration in patients with APACHEⅡscore>23(7 cases)was significantly lower than that in those with APACHEⅡscore 12-23(23 cases,μmol/L:5.33±1.72 vs. 10.02±5.24,t=2.300,P=0.001). Serum carnitine concentration in patients with serum total bilirubin(TBil)>19μmol/L(9 cases)was significantly lower than that in those with TBil≤19μmol/L(21 cases,μmol/L:5.54±2.70 vs. 9.84±5.08,t=2.750,P=0.014). Serum carnitine concentration was negatively correlated with the APACHEⅡscore and the TBil(r=-0.387,P=0.035;r=-0.346,P=0.048). During the 5-day observation period,enteral feeding amount〔(5 134±1 173)mL〕was positively correlated with serum carnitine concentration(r=0.430,P=0.022). In 30 critical patients,the incidence of abdominal distension was 40.0%(12/30),and the serum carnitine concentration of patients with abdominal distension was lower compared with that of patients without abdominal distension(μmol/L:7.83±4.98 vs. 9.12±5.35,t=0.707,P=0.383). The incidence of diarrhea was 26.7%(8/30),and the serum carnitine concentration of diarrhea patients was lower compared with that of patients without diarrhea(μmol/L:8.27±5.78 vs. 9.73±4.78,t=0.607,P=0.576). The mean length of hospital stay was(34.72±16.66)days. The serum carnitine concentrations in patients with hospital stay≥45 days (8 cases)were lower compared with those in those<45 days(22 cases,μmol/L:5.71±3.23 vs. 9.95±5.26,t=1.627,P=0.020). No correlation was found between serum carnitine concentrations and the hospital stay(r=-0.165, P=0.385). The length of ICU stay was(18.60±10.72)days. Serum carnitine concentration in patients with the length of ICU stay>7 days(27 cases)was slightly lower than that in those with the length of ICU stay≤7 days (3 cases,μmol/L:8.44±5.00 vs. 13.24±3.65,t=1.610,P=0.119). No correlation was found between serum carnitine concentrations and the length of ICU stay(r=-0.019,P= 0.293). In-hospital mortality was 26.67%(8/30). No significant difference in serum carnitine concentrations was found between the death group and the survival group(μmol/L:12.24±6.52 vs. 7.72±3.91,t=-1.846,P=0.098). No correlation was found between serum carnitine concentrations and in-hospital mortality(r=0.340,P=0.066). Conclusions Carnitine deficiency is significant in critically ill patients,and it is correlated with disease severity and serum TBil. The total amount of lenteral feeding was lower,and hospital stay was prolonged in critically ill patients with low serum carnitine level.

Objective To compare Luminex vs.ELISA methods in detecting HLA antibodies in kidney transplant recipients and their relation to acute rejection.Method Blood samples from 34 kidney transplant recipients were collected and the HLA antibodies were detected by both Luminex and ELISA methods.The sensitivity and specificity of both methods for predicting the development of acute rejection were analyzed.Results Fourteen recipients (14/34,41.17％) positive for HLA class Ⅰ antibodies were detected by using Luminex method,whereas only 1 case (1/34,2.9％) was detected with positive HLA class Ⅰ antibodies by ELISA method (P＜0.05).Similarly,13 recipients (13/34,38.24％) positive for HLA class Ⅱ antibodies were detected by using Luminex method,whereas the positive rate of HLA class Ⅱ antibodies by using ELISA method was 8.8％ (3/34,P＜0.05).The sensitivity and specificity of Luminex method for predicting the acute rejection were 80％ and 92.3％ respectively,in comparison to 30％ and 77.4％ respectively by ELISA method.Conclusion Compared to the traditional ELISA-based method,Luminex method has a better sensitivity and specificity for predicting the development of acute rejection.

Objective To analyze the clinical application of donor specific antibodies (DSAs) detected by a single antigen Luminex virtual crossmatch,and to discuss the treatment of DSA and the impact of DSA on renal function.Methods Serum from living-relative renal recipients before and after transplantation was investigated using a Luminex single antigen assay.The relation between DSA and renal acute rejection as well as renal function was analyzed.Results A total of 30 patients and 173 serum samples were tested,including 47 serum samples before transplantation,and 126 after transplantation.DSA was positive in one patient before transplantation,and 8 patients after transplantation.Three of the patients positive for DSA were treated by Bortezomib,3 by addition of MMF,2 by addition of CNI,1 by addition of Sirolimus.The MFI of DSA in one of the patients treated by Bortezomib was decreased to below 1000,while that in the other two decreased by more than 50 ％.The renal eGFR at the time with and without DSA was (1.50 ± 0.59) and (1.23 ± 0.38)ml/s respectively (P＜0.05).Conclusion Dynamic monitoring of single bead antigen antibody DSA conduces to direct the adjustment of immunosuppressant.The appearance of DSA contributes to the declination of renal function.Application of Bortezomib decreased the MFI of DSA.

Objective To detect de novo development of anti-HLA antibodies after renal transplantation, and to investigate their influence on graft function. Methods 384 kidney recipients,who were negative for anti-HLA antibody before transplantation, were monitored for anti-HLA antibodies over a period of 3-96 months, and a sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HLA antibodies. HLA antibody ＞10 % was defined as positive levels. Results Among 384 recipients tested, 318 recipients (82. 8 %) were negative for anti-HLA antibody after transplantation; 66 recipients (17. 2 %) developed de novo HLA antibodies, 3 recipients with HLA class Ⅰ, 61 with HLA class Ⅱ, 2 with both HLA class Ⅰ and Ⅱ. According to amino acid residue matching, 7 cases developed de novo antibodies among 92 recipients with 0 HLA-DR mismatches,compared with 59 cases among 292 recipients with 1-2 mismatches, which showed significant difference between two groups (P＜0. 01 ). 87. 4 % (278/318) recipients negative for HLA antibodies after transplantation achieved good graft function, in comparison with 65. 2 % (43/66) recipients positive for HLA antibodies (P＜0. 05). Conclusion De novo production of HLA antibodies posttransplantation may be closely associated with HLA-DR mismatch. De novo HLA antibodies posttransplantation might damage graft function and reduce graft survival rate. The detection of de novo development of anti-HLA antibodies after renal transplantation has clinical significance for assessing renal allograft function.