This study aims to investigate the effect of γ-Secretase Inhibitor DAPT, (N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester), on the differentiation of neural precursor cells and the production of neurons in the neural precursor cell line GE6. GE6 was cultured in medium with 4 μmol/L DAPT added as the experimental group and the untreated medium separately as the control group. After 4 days of differentiation, we carried out the following experiments. We used immuno-fluorescent staining to observe the ratio of Tuj1, GFAP and O4 positive cells. We also used qRT-PCR to detect the effect of the DAPT on Tuj1 and GFAP mRNA transcription in the GE6. The results of immuno-fluorescent staining indicated that the Tuj1 ratio of experimental group was higher compared to that of the control group, but the GFAP and O4 ratio of experimental group was lower than that of the control group. The differences were statistically significant (P < 0.05). The result of qRT-PCR was in accordance with immunofluorescent staining results. It was well concluded that DAPT could promote the neurogenic differentiation of neural precursor cell line rather than leading to gliogenic differentiation. More neurons could be obtained for transplantation with the addition of DAPT.
Amyloid Precursor Protein Secretases ; antagonists & inhibitors ; Animals ; Cell Differentiation ; drug effects ; Cell Line ; Dipeptides ; pharmacology ; Neural Stem Cells ; drug effects ; Neurons ; Rats

The purpose of this study was to identify specific microRNAs (miRNAs) during differentiation and maturation of interneurons and to predict their possible functions by analyzing the expression of miRNAs during in vitro differentiation of the rat interneuron precursor cell line GE6. In the experiment, the interneuron precursor cell line GE6 was cultured under three different conditions, i. e. the first was that had not added growth factors and the normal differentiation cultured for 4 days (Ge6_4d); the second was that cultured with bone morphogenetic protein-2 (BMP2) for 4 days (Ge6_bmp2); and the third was that cultured with sonic hedgehog (SHH) for 4 days (Ge6_ shh). In addition, another group of undifferentiated GE6 (Ge6_u) was applied as a control. We found in this study that the expression levels of a large number of miRNAs changed significantly during GE6 differentiation. The expression levels of miR-710, miR-290-5p and miR-3473 increased in the GE6 cells with secreted factor BMP2. These miRNAs may play important regulatory roles during interneuron differentiation.
Animals ; Bone Morphogenetic Protein 2 ; chemistry ; Cell Differentiation ; Cell Line ; Hedgehog Proteins ; chemistry ; Interneurons ; cytology ; metabolism ; MicroRNAs ; metabolism ; Rats

Background/Aims: To evaluate temporal trends of atrial fibrillation (AF) prevalence in critically ill patients who received prolonged mechanical ventilation (MV) in the United States. Methods: We used the 2008 to 2014 National Inpatient Sample to compute the weighted prevalence of AF among hospitalized adult patients on prolonged MV. We used multivariable-adjusted models to evaluate the association of AF with clinical factors, in-hospital mortality, hospitalization cost, and length of stay (LOS). Results: We identified 2,578,165 patients who received prolonged MV (21.27% of AF patients). The prevalence of AF increased from 14.63% in 2008 to 24.43% in 2014 (p for trend < 0.0001). Amongst different phenotypes of critically ill patients, the prevalence of AF increased in patients with severe sepsis, asthma exacerbation, congestive heart failure exacerbation, acute stroke, and cardiac arrest. Older age, male sex, white race, medicare access, higher income, urban teaching hospital setting, and Western region were associated with a higher prevalence of AF. AF in critical illness was a risk factor for in-hospital death (odds ratio, 1.13; 95% confidence interval, 1.11 to 1.15), but in-hospital mortality in critically ill patients with AF decreased from 11.6% to 8.3%. AF was linked to prolonged LOS (2%, p < 0.0001) and high hospitalization cost (4%, p < 0.0001). LOS (–1%, p < 0.0001) and hospitalization cost (–4%, p < 0.0001) decreased yearly. Conclusions The prevalence of comorbid AF is increasing, particularly in older patients. AF may lead to poorer prognosis, and high-quality intensive care is imperative for this population.

OBJECTIVETo optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis.

METHODFour methods, i. e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification.

RESULTThe total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test.

CONCLUSIONThe bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.

Bentonite ; chemistry ; DNA, Complementary ; analysis ; Electrophoresis ; Fritillaria ; genetics ; Guanidines ; chemistry ; Isothiocyanates ; chemistry ; Phenol ; chemistry ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; RNA, Plant ; analysis ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium Dodecyl Sulfate ; chemistry ; Time Factors

Objective To investigate the serum levels of vitamins and antioxidative indexes as well as theircorrelations with nonalcoholic fatty liver disease (NAFLD),and to analyze the influence of different detection methods on same index detection result.Methods Eighty NAFLD patients diagnosed by ultrasound were chosen for detecting the indexes,including the levels of serum RBP,vitamin A (VA),vitamin D (VD,ELISA and enzyme donor competition method),vitamin E (VE),vitamin B12 (VB12),and antioxidative indexes,including malondialdehyde (MDA),superoxide dismutase (SOD,pyrogallol substrate method and WST-1 method) and reduced glutathione (GSH).Results Compared with the normal reference values,the deficiency rate of serum RBP,VA,VD,VE and SOD in NAFLD patients were 11.6％,38.4％,7.9％,86.0％ and 27.9％ respectively.The serum GSH level in the NAFLD group was lower than that in the healthy control group (P＜0.01).Detecting serum VD and SOD by different detection method found that the difference among different detection methods had statistical significance (P＜0.01).Conclusion The levels of serum VA,VE,GSH and SOD in NAFLD patients are significantly reduced.These detection indicators have the important significance for clinical diagnosis of NAFLD.

Objective:To investigate the correlation between the prognosis of patients infected with BK virus after renal transplantation and their peripheral blood related indexes.Methods:131 patients from the Renal Transplantation Department of the Second Xiangya Hospital of Central South University who underwent renal transplantation and firstly infected with BK virus after the surgery during the period from August 2018 to August 2021 were retrospectively analyzed. 93 males (71.0%) and 38 females (29.0%). The average age was (37.5±11.3) years old. 109 cases underwent cadaveric kidney transplant (83.2%) and 22 cases underwent relatives kidney transplant (16.8%). The onset time of the first infection with BK virus after renal transplantation was (188.7±16.6) days, and the serum creatinine was (127.5±39.5) μmol/L. 25 patients (19.1%)infected with BK virus were positive in blood and urine at the same time, and 106 patients (80.9%)infected with BK virus were positive only in urine. Among 131 patients infected with BK virus, 70 patients were treated by lowering the blood concentration of tacrolimus to enhance immunity, 12 patients were treated by switching tacrolimus to cyclosporine, and 49 patients had incomplete follow-up data. The DNA load of BK virus in 25 patients [5.6(2.4, 12.3)×10 3copies/ml] positive in blood, white blood cell count(WBC)(5.8±2.0)×10 9/L, hemoglobin(Hb)(122.0±22.4)g/L, platelet count(PLT)(187.1±63.1)×10 9/L, neutrophil count(NEUT)(3.9±1.7)×10 9/L, lymphocyte count(LYM)(1.5±0.8)×10 9/L, monocyte count(MONO)(0.4±0.2)×10 9/L, neutrophil to lymphocyte ratio(NLR)2.2(1.7, 3.5), derived neutrophil to lymphocyte ratio(dNLR)1.7(1.3, 2.6), platelet to lymphocyte ratio(PLR)121.3(86.3, 227.3), monocyte to lymphocyte ratio(MLR)0.2(0.1, 0.4) and lymphocyte to monocyte ratio(LMR)4.7±2.6. The DNA load of BK virus in 106 patients [20.4(0.4, 2 570.0)×10 5copies/ml] positive in urine, WBC 6.6(4.8, 9.1)×10 9/L, Hb(129.0±24.5)g/L, PLT 188.0(147.3, 226.5)×10 9/L, NEUT 4.6(3.0, 6.6)×10 9/L, LYM(1.7±0.8)×10 9/L, MONO 0.4(0.3, 0.5)×10 9/L, NLR 2.8(1.9, 3.9), dNLR 2.1(1.5, 3.0), PLR 120.5(87.0, 163.2), MLR 0.2(0.1, 0.4), LMR 4.5(2.8, 6.7). 70 patients infected with BK virus treated by lowering the blood concentration of tacrolimus were divided into BK virus rise group and BK virus decline group according to the change of BK virus DNA load in blood and urine before and after treatment (the grouping principle of this study gives priority to the change of BK virus DNA load in blood, followed by the change of BK virus DNA load in urine). The WBC, Hb, PLT, NEUT, LYM, MONO, NLR, dNLR, PLR, MLR, LMR, tacrolimus blood concentration and change difference, blood creatinine and change difference were analysed between two groups. Results:The BK virus DNA load in 25 patients positive in blood was correlated with NLR and dNLR ( r=0.5062, P=0.0098; r=0.5738, P=0.0027), and there was no correlation between the BK virus DNA load in blood with the WBC ( r=-0.0185, P=0.9302), Hb ( r=0.0912, P=0.6646), PLT ( r=-0.3931, P=0.0519), NEUT ( r=0.2438, P=0.2401), LYM ( r=-0.3035, P=0.1402), MONO ( r=-0.3279, P=0.1096), PLR( r=0.1054, P=0.6161), MLR( r=0.0738, P=0.7257), LMR( r=-0.0738, P=0.7257). There was no correlation between the BK virus DNA load in 106 patients positive in urine and WBC( r=0.0222, P=0.8209), Hb( r=-0.0323, P=0.7423), PLT( r=0.0847, P=0.3881), NEUT( r=0.0417, P=0.6713), LYM( r=0.0010, P=0.9916), MONO( r=0.0224, P=0.8196, NLR( r=0.0170, P=0.8623), dNLR ( r=-0.0013, P=0.9892), PLR( r=0.0387, P=0.6934), MLR( r=-0.0070, P=0.9433)and LMR( r=0.0070, P=0.9433). As for 70 patients infected with BK virus, there were 37 patients in the BK virus rise group and 33 patients in the BK virus decline group. In the two groups, age [(38.4±12.0)years old and(39.0±9.0)years old], gender [male /female: (23/14) cases and(27/6)cases], blood type [A+ /B+ /AB+ : (22/13/20)cases and (26/6/1)cases], donation type [relatives donnation/cadaveric donation: (29/8)cases and (27/60)cases], blood creatinine(after treatment)[123.0(98.4, 140.5)μmol/L and 132.0(107.1, 162.4)μmol/L] and change difference before and after treatment [0(-15.7, 10.5)μmol/L and -2.0(-9.1, 15.0)μmol/L], tacrolimus blood concentration (after treatment)[(6.7±2.0)ng/ml and(6.5±1.5)ng /ml] and tacrolimus concentration change difference [-1.4(-3.8, 0.6)ng/ml and -1.2(-2.2, 1.3)ng/ml] had no significant difference( P<0.05). The MONO of the two groups was statistically different [0.3(0.2, 0.5)×10 9/L and 0.4(0.3, 0.6)×10 9/L, P=0.033], and there was no difference between the two groups in WBC[6.6(4.1, 8.8)×10 9/L and 6.8(5.4, 8.9)×10 9/L], Hb[(133.2±25.3)g/L and(131.6±20.6)g/L], PLT[185.0(151.0, 231.5)×10 9/L and 196.0(149.0, 234.0)×10 9/L], NEUT[4.3(2.4, 6.4)× 10 9/L and 4.2(3.1, 5.5)×10 9/L], LYM[1.7(1.1, 2.2)×10 9/L and 1.8(1.1, 2.3)×10 9/L], NLR[2.5(1.9, 3.8)and 2.4(1.9, 3.7)], dNLR [2.0(1.5, 2.8)and 1.9(1.4, 2.5)], PLR [114.9(85.1, 159.4)and 111.3(77.1, 159.6)], LMR(4.6±2.6 and 5.2±2.4), MLR[0.2(0.2, 0.4)and 0.2(0.2, 0.4)]( P<0.05). Conclusions:There is a positive correlation between the blood BK virus DNA load and NLR, dNLR in renal transplant recipients infected with BK virus. The rise of MONO correlates with good prognosis of BK virus.

The effect of living kidney transplantation between identical twins is satisfied, but it is rarely reported. From October 2019 to February 2021, two recipients received kidney transplantation from their twin sisters in the Second Xiangya Hospital of Central South University. The primary disease of the two recipients was acute glomerulonephritis in 1 case and diabetic nephropathy in 1 case. Two recipients received tacrolimus/cyclosporine+ mortemycophenol ester+ methylprednisolone after surgery. The patients were followed up for 3.0 and 1.5 years, respectively, with renal function recovering well.

Objective:To explore the clinical efficacy of adult donor dual kidney transplantation.Methods:Retrospective analysis of case data of 13 adult donor kidney dual kidney transplantation (DKT) performed in the The Second Xiangya Hospital of Central South University from September 2016 to December 2020. For 13 donors, the average age and BMI were (53.5±12.4)years and (24.3±2.8) kg/m 2, respectively. Their mean Serum creatinine (SCr) at admission and before procurement was (132.9±54.1)and (228.7±112.4)μmol/L, respectively. 3 of them had diabetes mellitus history, and 8 had hypertension history. 11 met the United Network for Organ Sharing (UNOS) DKT criteria and 6 met Remuzzi score DKT criteria. For 13 recipients, the average age and BMI were (39.3±8.9)years and (20.2±2.4)kg/m 2, respectively. All of them received ABO blood type-matched kidney transplants. 2 of them had their grafts transplanted in the bilateral iliac. In 12 cases, the grafts filled rapidly and urinated immediately when opening blood flow. In 1 case, the grafts were dark in color and vascular showed weak pulsation after opening blood flow. The time to recovery of perioperative graft function (from the day of surgery to the natural reduction of SCr to the normal range 44-133μmol/L), the occurrence of delayed graft function (DGF), acute rejection (AR), ureteral and surgical incision complications, as well as the recipients’ final follow-up SCr, eGFR, urinary protein, and grafts outcome were observed. Risk factors affecting outcomes were assessed by univariate logistic regression analysis. Results:The SCr dropped to the normal range at discharge in 10 recipients, and the average recovery time was (13.8±13.0) days. In other 3 cases SCr at discharge were 300.0, 149.0, 152.5μmol/L. 4 cases had DGF, 4 had AR, 1 experienced urinary fistula, and 1 experienced incisional dehiscence, which were treated with anti-rejection, J-tube implantation, continuous catheterization to maintain bladder void, secondary suturing, respectively. The follow-up time ranged from 4 to 54 months, with a median of 28(15.5, 31.0) months. At the final follow-up time, 10 cases had good graft function, 2 suffered impaired kidney function, and 1 experienced graft failure. The average SCr and eGFR except for graft failure patient were (144.2±101.3)μmol/L and (52.9±21.2)ml/min, respectively. 4 had positive urine protein. Univariate logistic regression analysis showed that donor age, BMI, history of diabetes mellitus and hypertension, and SCr were not significantly correlated with recipients’ DGF and graft impairment ( P>0.05), and due to the small sample size, multifactorial logistic regression analysis was not performed. Conclusion:The short to medium-term effects of adult donor DKT coule be safe and feasible.

Objective To investigate the antigen presentation characteristics of dendritic cells(DC)and B cells in cardiac grafts.Methods The heart of BALB/c mice was transplanted into the abdominal cavity of C57BL/6J mice.CD45+cells in the heart graft were extracted and sorted by flow cytometry at postoperative 5 d,and single cell RNA sequencing was performed.Taking DC and B cell subsets in cardiac grafts as the main study cells,the changing trend,antigen presenting ability and intercellular communication with T cells after heart transplantation were analyzed by bioinformatics analysis and flow cytometry.Gene ontology(GO)function enrichment difference analysis was adopted to prove the specific function and the reliability annotation of cell subsets.Results Germinal center-like B cell(GC-L B)was the B cell subset with the largest increase in quantity during the acute rejection phase,accounting for 87％.Classical DC(cDC)2 was the only DC subset with a significant increase in quantity during acute rejection of heart transplantation,accounting for 44％of DC subset,and it occupied the highest communication intensity with T cells after heart transplantation.Mononucleated DC(moDC)and memory B cell(MBC)were the main transmitters of T cell input signals in non-transplanted hearts,whereas transformed into cDC2 and GC-L B during the acute rejection phase.Among them,MBC and GC-L B were the main sources of T cell input signals in non-transplanted hearts and heart grafts.Conclusions Compared with DC,B cells occupy a higher number and weight in the intercellular communication with T cells in non-transplanted hearts and heart grafts,prompting that the antigen presenting activity of B cells is more active and stronger than DC in the early stage of acute rejection of heart transplantation.

Objective: To evaluate and describe the changes of core muscle groups based on DAVID spine biomechanics training system in ankylosing spondylitis (AS) patients. Methods: The clinical data of 100 patients of AS and 31 healthy controls were collected. Clinical symptoms, Bath ankylosing spondylitis disease activity index (BASDAI), Bath ankylosing spondylitis function index (BASFI), Bath ankylosing spondylitis measurement index (BASMI), ankylosing spondylitis disease activity (ASDAS), and simultaneous detection of DAVID spine biomechanics training system, simple core muscle fitness test: Eight-grade abdominal bridge, PLANK exercise (flat support), Abdominal static muscle endurance test, Back static muscle endurance test were compared using t-test analysis and spearman correlation analysis. Results: ① Between AS and healthy male control o group, there were significant differences of spinal mobility in forward flexion, right rotation, left rotation (42±13 vs 48±1, 52±14 vs 69±12, 52±13 vs 58±11; all P values <0.05); and significant differences of spinal muscle strength in forward bending force, right rotation force, left rotation force, right bending force (103±42 vs 146±17, 87±34 vs 104±13, 80±35 vs 101±13, 161±55 vs 186±19; all P values <0.05), and significant differences in the left/right rotational force (1.17±0.21 vs 1.02±0.111, P<0.05) of spine balance strength comparison.② Between AS and healthy controls of female group, there were differences in forward bending force (49±23 vs 77±10, P<0.05) of spinal muscle strength; and significant differences in forward bending/backward extension strength, left and right rotation strength (0.32±0.11 vs 0.58±0.21, 1.29±0.21 vs1.03±0.11, all P values <0.05) of spine balance strength; ③ In AS group, the spinal mobility was correlated with age (Rear extension r=-0.28, right flexion r=-0.268, left flexion r=-0.404, right rotation r=-0.367, left rotation r=-0.235; all P values <0.05), course of disease (Rear extension r=-0.354, forward flexion r=-0.283, right flexion r=-0.204, left flexion r=-0.284, right rotation r=-0.339, left rotation r=-0.23; all P values <0.05), body mass index (BMI) (Rear extension r=-0.23, forward flexion r=-0.288, right flexion r=-0.22, left flexion r=-0.201, right rotation r=-0.26, left rotation r=-0.29; all P values <0.05), sacroiliac joint stage(Rear extension r=-0.375, forward flexion r=-0.446, right flexion r=-0.331, left flexion r=-0.367, right rotation r=-0.368, left rotation r=-0.314; all P values <0.05) and BASDAI (Rear extension r=-0.381, forward flexion r=-0.374; all P values <0.05). Spinal muscle strength was correlated with gender (Posterior extensor force r=0.344, flexor force r=0.507, right rotation force r=0.376, left rotation force r=0.399, right flexion force r=0.433, left flexion force r=0.445; all P values <0.05); the left/right spine rotation strength was correlated with gender (r=0.271, P<0.05). ④ In the simple core muscle fitness test, eight-grade abdominal bridge was correlated with spinal muscle strength (Rear extension force r=0.234, right rotation r=0.290, left rotation r=0.219, right flexion r=0.35, left flexion r=0.327; all P values <0.05); PLANK exercise was correlated with spinal muscle strength (Rear extension force r=0.234, right rotation r=0.290, left rotation r=0.219, right flexion r=0.35, left flexion r=0.327; all P values <0.05); abdominal static muscle endurance test was correlated with forward flexion strength (r=0.341, P<0.05); back static muscle endurance test was correlated with spinal mobility (Rear extension r=0.262, forward flexion r=0.23, right rotation r=0.455, left rotation r=0.426, right flexion r=0.387, left flexion r=0.46; all P values <0.05); correlated with spine strength (right flexion r=0.256, left flexion r=0.272; all P values <0.05). Conclusion Compared with healthy people, AS patients have decreased activity, strength and balance of spinal core muscle. There are significant decline in spinal mobility and muscle strength of male AS patients and muscle imbalance of female AS patients. Simple core muscle fitness test could be used in clinic to measure the changes of AS patients'core muscle group.