Objective To explore the relationship between the index of SPECT myocardial viability teat of the left ventricle and long-term cardiovascular adverse events following the coronary artery bypass graft.Methods Clinical data were selected from the coronary-heart-disease patients with CABG and the isotopic SPECT test of myocardial viability prior to the operation in the Fuwai Cardiovascular Hospital,the Chinese Academy of Medical Science from January 1,1999 to December 31,2005.Total 709 patients were included in this study.Regular follow-up in patients was performed following operation.Investigate the nuclear medicine documents of the patients above;make the semi-quantitative scores of myocardial viability prior to operation by the 17-segment method of ventricle.Through the COX multi-factor analysis of the statistical methods to explore the relationship between the index of the myocardial viability and the long-term cardiac adverse events selected in advance.Results The mean duration of follow-up was(3.43±2.42)years.The COX multi-analysis revealed that the total score of left ventricular myocardial viability is the independent impact factors for long term cardiac death,long terrm re-hospitalization rate and long term composite end points events.The cut off values of total score of left ventricular myocardial viability for long term cardiac death,long term re-hospitalization rate and long term composite end points events is 15,9,13 respectively.Conclusion The total score of left ventricular myocardial viability of SPECT is independently associated with long-term events,and the cut values of myocardial viability total score for long-term cardiac death,re-hospitalization and composite MACE events are 15,9,and 13 respectively.For the groups with total scores above and below the cut off values,there is significant difference of long term cardiac events risk between groups.

Objective:To explore the mechanism of herb-partitioned moxibustion in relieving rat intestinal inflammation by focusing on the neutrophil extracellular traps(NETs)in Crohn disease(CD)development. Methods:Rats were randomly divided into a normal group,a model group,a herb-partitioned moxibustion group,and a mesalazine group.The CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid except for rats in the normal group.Rats in the normal group and model group did not receive any treatment but had the same fixation as the other groups.Rats in the herb-partitioned moxibustion group received herb-partitioned moxibustion at Qihai(CV6)and bilateral Tianshu(ST25).Rats in the mesalazine group received intragastric administration of mesalazine enteric-coated tablets.The general situation of rats in each group was recorded,and the histopathological changes in the colon were observed and scored by hematoxylin-eosin staining.The serum concentrations of NETs DNA(NETs-DNA),neutrophil elastase(NE)-DNA,and myeloperoxidase(MPO)-DNA were detected by ABC enzyme-linked immunosorbent assay,and the citrullinated histone 3(citH3),MPO,and NE protein and mRNA expression levels in rat colon tissue were observed by immunofluorescence and real-time quantitative polymerase chain reaction. Results:Compared with the normal group,the mucosal ulcer reached the muscularis,the epithelium was incomplete,the goblet cells decreased obviously with significant inflammatory cell infiltration in the colon;the colonic mucosa damage index(CMDI)score increased significantly(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations increased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue increased significantly in the model group(P<0.01 or P<0.05).Compared with the model group,the mucosal epithelium in the herb-partitioned moxibustion group and the mesalazine group was repaired and the goblet cells increased with a few infiltrating inflammatory cells in the colon;the CMDI score decreased(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations decreased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue was down-regulated(P<0.01 or P<0.05). Conclusion:Herb-partitioned moxibustion reduced the serum NETs complex and inhibited the protein and mRNA expression of NETs complex in the colon tissue,which may be one mechanism of herb-partitioned moxibustion in relieving colon mucosal inflammation in CD.

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo construct human renal cell carcinoma patient-specific full-length antibody library using mammalian cell surface display technique.

METHODSPeripheral blood mononuclear cells (PBMC) were isolated from patients with renal cell carcinoma. The repertoires of kappa light chain (LCkappa) and heavy chain variable region (VH) of antibody were amplified by RT-PCR. The LCkappa and VH libraries were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO10 to construct the renal cell carcinoma patient-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length human antibodies expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of renal cell carcinoma-specific antibody kappa light chain (LCkappa) and heavy chain (IgG1) were constructed. The expression of the full-length human antibodies on the surface of 293T cell was confirmed by flow cytometry. The libraries showed an expressible combinatory diversity of 7.5x10(10).

CONCLUSIONThe expressible antibody library provides a useful platform for screening of renal cell carcinoma-specific antibodies.

Amino Acid Sequence ; Antibodies, Neoplasm ; immunology ; Antibody Specificity ; Carcinoma, Renal Cell ; immunology ; Humans ; Kidney Neoplasms ; immunology ; Molecular Sequence Data ; Peptide Library

Objective:To establish and evaluate a new real-time quality control method that can identify the random errors by using the backpropagation neural network (BPNN) algorithm and taking blood glucose test as an example.Methods:A total of 219 000 blood glucose results measured by Siemens advia 2 400 analytical system from January 2019 to July 2020 and derived from Laboratory Information System of Beijing Chaoyang Hospital Laboratory Department was regarded as the unbiased data of our study. Six deviations with different sizes were introduced to generate the corresponding biased data. With each biased data, BPNN and MovSD algorithms were used and tested, and then evaluated by traceability method and clinical method.Results:For BPNN algorithm, the block size was pre-set to 10 and the false-positive rate in all biases was within 0.1%. For MovSD, however, the optimal block size and exclusive limit were 150 and 10% separately and its false-positive rate in all biases was 0.38%, which was 0.28% higher than BPNN. Especially, for the least two error factors of 0.5 and 1, all the random errors were not detected by MovSD; for the error factor larger than 1, random errors could be detected by MovSD but the MNPed was higher than that of BPNN under all deviations. The difference was up to 91.67 times. 460 000 reference data were produced by traceability procedure. The uncertainty of BPNN algorithm evaluated by these reference data was only 0.078%.Conclusion:A real-time quality control method based on BPNN algorithm was successfully established to identify random errors in analytical phase, which was more efficient than MovSD method and provided a new idea and method for the identification of random errors in clinical practice.

【Objective】Diabetes mellitus is a risk equivalent for coronary heart disease. This retrospective study was designed to investigate the risk factors of the progression of coronary lesions in patients with type 2 diabetes（T2DM）and Non- diabetes Mellitus（NDM）.【Methods】 526 patients with T2DM and 425 patients with NDM at the Third Affiliated Hospital of Sun Yat-sen University between March 2001 and January 2017 who underwent coronary imaging studies（coronary angiography or coronary CTA）twice during the same period were enrolled. The effects of cardiovascular risk factors on the progression of coronary lesions were analyzed in parallel in these two types of patients.【Results】Risk factors of the progression of coronary lesions in T2DM patients included smoking（OR = 1.836，95% CI：1.030~3.371，P = 0.04），Lp（a） ［OR = 1.001，95% CI：1.000~1.002，P = 0.004（baseline）；OR = 1.001，95% CI：1.000~1.002，P = 0.009（re-examined）］，HbA1c leve［l OR = 1.471，95% CI：1.030~2.100，P = 0.034（re-examined）］，uncontrolled LDL-C（OR = 1.882，95% CI：1.091~3.245，P = 0.023），TC［OR = 2.029，95% CI：1.028~4.008，P = 0.041（re-examined）］and low HDL-C ［OR = 0.017，95% CI：0.040~0.729，P = 0.017（re-examined）］. Comparative risk factors in NDM included BMI［OR =1.746，95%CI：2.462~2.712，P = 0.026（baseline）；OR = 0.001，95%CI：0~0.394，P = 0.025（re-examined）］，uncontrolled LDL-C（OR = 2.875，95%CI：1.669~4.952，P < 0.001）and low ApoA［OR = 0.282，95%CI：0.082~0.971，P = 0.045 （baseline）；OR = 0.117，95%CI：0.038~0.835，P = 0.029（re-examined）］. Lowest level of progression was found in the group with HbA1c<6.5%［0（0~3.4）points/year vs 0.3（0~3.0）points/year vs 1.0（0~5.1）points/year，P = 0.049. 0（-0.4~2.7）points/year vs 0.6（0~4.0）points/year vs 0.9（0~4.2）points/year，P=0.029］in T2DM patients.【Conclusion】Except for achievement of LDL- C goals，there might be some differences in risk factors for progression of coronary lesions between T2DM and NDM patients. Smoking，Lp（a），TC，HDL- C and control levels of HbA1c are independent predictors in T2DM as well as BMI and ApoA in NDM. Lowering HbA1c to less than 6.5% may delay progression of lesion.

Humans ; Hydrogen-Ion Concentration ; Malassezia ; growth & development ; physiology ; Staphylococcus epidermidis ; growth & development ; physiology

Objective To determine the influences of Mannose binding protein (MBP) gene polymorphisms on HBV DNA loads and on the progression of liver disease in patients with chronic HBV infeclion.Method The Codons on 54 MBP gene polymorphisms and HBV DNA loads in a cohort of 395 patients with chronic HBV infection,including 244 with chronic hepatitis B (CHB),151 with liver cirrhosis(LC) and 88 normal controls were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and fluorescent quantitative PCR (FQ-PCR).Result The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC in CHB group showed no significant differences comparing to the normal control group ( P ＞ 0.05 ).The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC on CHB group (severe),compensation phase of LC group and decompensation phase of LC group were higher than those in the normal control group (P ＜ 0.05 ),the genetic polymorphism of decompensation of LC was 36.5 ％,highest of all.The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC of patients with chronic HBV infection were not changed with the differences of HBV-DNA loads.Conclusion The codes on 54 MBP gene polymorphisms is not closely related to HBV DNA loads,but was associated with the progression of hepatitis B infection.

Objective To establish a reverse remodeling heart model in rats and observe collagen and TGF-β expression and relevant rmicroRNAs changes during reverse remodeling. Methods Lewis rats were divided into four groups including sham(NL, n = 10), abdominal aortic constriction(AAC, n = 10),heterotopic transplantation of abdominal aortic constriction(AAC-HT, n = 9)and heterotopic transplantation of normal heart(HT, n = 8). Left ventricular wall thickness and LV cavity were measured by echocardiography. The cardiomyocyte cross-sectional area(CSA)was determined on HE stained sections.Immunohistochemical and qRT-PCR were used to detect collagen and TGF-β expressions. miRNAs were detected by MicroRNA microarray. Results Heart weight, left ventricular wall thickness and CSA were significantly increased in AAC hearts compared to those in the NL and AAC-HT hearts. The collagen and TGF-β were increased in AAC hearts and further increased in AAC-HT hearts. miRNA microarray evidenced more than two folds changes on 82 miRNAs compared to NL(10 in AAC, 32 in AAC-HT and 40 in HT).Conclusion Rat abdominal aortic constriction and heterotopic transplantation could be used as a reverse remodeling heart model and significant collagen and TGF-β as well microRNA expression changes were evidenced in this model.

Major advances have been made over the past few decades in identifying and managing disorders of consciousness (DOC) in patients with acquired brain injury (ABI), bringing the transformation from a conceptualized definition to a complex clinical scenario worthy of scientific exploration. Given the continuously-evolving framework of precision medicine that integrates valuable behavioral assessment tools, sophisticated neuroimaging, and electrophysiological techniques, a considerably higher diagnostic accuracy rate of DOC may now be reached. During the treatment of patients with DOC, a variety of intervention methods are available, including amantadine and transcranial direct current stimulation, which have both provided class II evidence, zolpidem, which is also of high quality, and non-invasive stimulation, which appears to be more encouraging than pharmacological therapy. However, heterogeneity is profoundly ingrained in study designs, and only rare schemes have been recommended by authoritative institutions. There is still a lack of an effective clinical protocol for managing patients with DOC following ABI. To advance future clinical studies on DOC, we present a comprehensive review of the progress in clinical identification and management as well as some challenges in the pathophysiology of DOC. We propose a preliminary clinical decision protocol, which could serve as an ideal reference tool for many medical institutions.
Humans ; Transcranial Direct Current Stimulation/methods* ; Consciousness Disorders/etiology* ; Brain Injuries/complications* ; Consciousness ; Neuroimaging