Objective To investigate the effects of rhG-CSF on the improvement of cognitive impairment and anti-apoptosis of Alzheimer disease (AD) induced by Aβ1-42 in rat. Methods Healthy male Wistarirats were randomly assigned to the Aβ group, treatment group and sham operation group. Aβ1-42 (10μg) was injected into bilateral hippocampus to create the rat model of AD. Rats in rhG-CSF group were subcutaneously subjected to 50μg/(kg · d) rhG-CSF for 5 days, while rats in Aβ group were subjected to normal saline. Morris water maze tests were done and expressions of caspase-3 protein were determined by immunohistochemical method on the 7th, 14th, 21st, and 28th day after administration. Results (1) The avoiding latent periods of rhG-CSF group ( ( 34. 33 ±6. 47 ) s, (42. 08 ± 6. 36 ) s, (46. 88 ± 7. 66 ) s, respectively ) were shorter than that of Ap group ((49.79 ±4.87)s, (50.25 ±6.81 )s, (51. 33 ±6.90)s, respectively). The percentages of swimming distances in the target quadrant in rhG-CSF group ( (41.00 ±7.62)% ,(43.33 ±8. 16)% ,(44. 67 ±8.07)% ,respectively) were increased comparing with Ap group((25.33 ±6.89)% , (23. 83 ±4.67)% ,(21.50 ±4.64)% ,respectively). The differences were all statistically significant (P<0.05). (2) Compared with sham operation group,the positive rate of caspase-3 protein in rat's hippocampus of Ap group significantly increased after injecting Aβ1-42 The positive rates of caspase-3 protein in rhG-CSF group on the 7th, 14th day ( (7. 93 ±6. 33) and (8. 83 ±5. 94) were lower than Aβ group ( ( 10.43 ±7. 16) and ( 11. 34 ± 5. 17 ) . The differences were statistically significant (P< 0.05). Conclusion RhG-CSF can improve the cognitive impairment of Alzheimer Disease (AD) induced by Aβ1-42 in rat, decrease the apoptosis of hippocampal neurons and delay the decline of its learning and memory ability to some extent.

Objective To investigate the spatiotemporal expression patterns and the relationship of α-sarcomeric actin(α-SCA) ,α-smooth muscle actin(α-SMA) and intermediate filament protein desmin with the maturation of the prenatal and the neonatal mouse hearts. Methods Serial sections of the embryo mouse and the neonatal mouse hearts were immunostained with antibodies against α-SCA, α-SMA and desmin. Results Ventricle and outflow tract of embryonic day(ED) 9 heart showed stronger expression of α-SCA and α-SMA, but desmin expression was lower. In the atrium, the expressions of α-SCA and α-SMA were restricted to the dorsal and ventral walls. In the sinus venosus, only a few weakly stained α-SCA positive cells were detected. No desmin expression was found in the atrium and sinus venosus. The expressions of α-SCA, α-SMA and desmin were increased to their highest level at ED 12. The higher expression of α-SCA remained to the postnatal stages. After ED 12, the expressions of α-SMA and desmin gradually decreased in different parts of the heart, but their expressions in the right ventricle persisted longer. After birth,desmin expression was mainly concentrated in the Z lines of I bands and intercalated disks. Conclusion The presence of spatiotemporal differences in the expression of α-SMA and desmin reveals regional differences in cardiomyocyte maturation in various parts of the embryonic mouse heart. The right ventricle shows a relatively slow pace of maturation. The α-SMA may contribute to a peristaltoid contraction pattern of the embryonic myocardium with a slow shortening speed, and a relatively higher level of desmin is required for the maturation of the sarcomere.

Objective To investigate the early development of the sinus venosus and the cardiac conduction system (CCS) of human embryonic hearts. Methods Serial transverse sections of 29 human embryonic hearts from Carnegie stage 10 to Carnegie stage 16 (C10-C16) were stained immunohistochemically with antibodies against α-smooth muscle actin(α-SMA),α-sarcomeric actin(α-SCA) and desmin ( DES ). Results During C12 and C13, the sinus venosus formed by confluence of systematic veins at the caudal end of the pericardial cavity could be recognized in the mesenchyme of primitive transverse septum. The mesenchymal cells of the sinus venosus gradually differentiated into α-SCA positive cardiocyocytes. At C14, the sinus venosus was within the pericardial cavity due to expansion of the pericardial cavity and incorporated into the right atrium. Differentiation of DES positive conductive cardiomyocyte was initiated in the right wall of atrio-ventricular canal of C10 embryonic heart and with the development, extended towards the myocardium of the interventricular sulcus to form His bundle, left and right bundle branches as well as the ventricular trabecular myocardium. In the atium, the strong expression of DES was first detected in the dorsal wall of C11 atrium. At C13, unique myocardial band showing α-SCA, α-SMA and DES expression in the left dorsal wall of the sinus venosus were found to be continuous with the basal wall of left atium and the dorsal wall of the atrio-ventricular canal, this band might be related to the development of conduction system from sinoatrial node to atrio-ventricular canal. During C14 to C16, primary conduction pathway of atria with strong DES expression was formed that extended from sinoatrial node along venous valves, DES positive myocardium in the dorsal and ventral walls of the atria to the right atrio-ventricular canal, respectively. Conclusions The mesenchyme of the primitive transverse septum is the heart forming field of human embryos responsible for formation of sinus venosus myocardium, cardiomyocytes are differentiated from mesenchymal cells in the primitive transverse septum and progressively added to the venous pole of the heart tube to form myocardial sinus venosus. The differentiation of CCS of the early human embryo initiates in the atrio-ventricular canal and develops gradually towards the arterial and venous poles of the heart tube. By C16, DES positive embryonic CCS can be clearly recognized morphologically.

Objective To investigate the origin of α-SMA positive cells in the outflow tract ridge of the embyonic mouse heart and to explore the ultrastructure change of the mesenchymal cells during the ridges fusion. Methods Sections of embryonic day 10(E10d) to E14d mouse embryonic hearts were stained by immunohistochemistry assay with monoclonal antibodies against α-smooth muscle actin (α-SMA), α-sarcomeric actin(α-SCA) and in situ hybridization method with PlexinA2 probe. The outflow tract ridges fusion was observed by transmission electron microscopy at E12.5d. Results From E10d to E11d, PlexinA2 positive cells were seen in the neural tube with mesenchymes around it, the aortic sac and aortic arch. These cells also migrated into the outflow tract ridge along the aortic sac wall and part of them expressed α-SMA. At E12d, PlexinA2 was expressed in the spinal ganglia, the pharyngeal mesenchyme, the aorto-pulmonary septum and the ascending aorta and pulmonary trunk. The septum showed α-SMA strongly positive. But only a few of α-SMA positive cells were observed in the ascending aorta and pulmonary trunk. At E12.5d, two clusters of condensed mesenchymal cells in the outflow tract ridges formed and began to express PlexinA2 weakly and α-SMA strongly. When the ridges began to fuse, the endothelial cells formed a cellular seam, which rapidly broke into pieces and disappeared and were replaced by the sparsed mesenchymal cells containing a few of microfilaments. Two clusters of condensed mesenchymal cells gradully moved to merge. It was noted that some mesenchymal cells contained plenty of microfilament bundles, mitochondria and focal contacts between them. Some mesenchymal cells only had a few of microfilaments and plasma membrane fusion was seen between them. Conclusionα-SMA positive cells in the outflow tract cushion may be derived from cardiac neural crest. The endothelial cells might participate in the fusion of the outflow tract ridges by endothelial-mesenchymal transformation. Mesenchymal cells of the condensed cell mass contain plenty of microfilament bundles and focal contacts or membrane fusion, which contribute to the ridges fusion.

Objective The purpose of this study was to investigate the expression of mitochondrial motility-related gene mirol and mitochondrial energy metabolism during an acute bout of prolonged exercise. Methods CS7 BL/6 mice underwent a moderate intensity treadmill running with the slope of 0° and at the speed of 13m/min, and they were randomly divided into 5 groups:resting group(R),exercise groups running for 30min(E30),60min(E60),90min (E90) and 120min(E120),respectively. Respiratory control index and ATP synthesis activity in isolated mitochondria were detected. Skeletal muscle H_2O_2 concentration,mirol mRNA expressions were also measured. Results (1)mirol mRNA contents were significantly increased in groups E30 to E120,as compared with the group R;and mirol mRNA expression increased 32.8%, 107.6%,63.8% and 44.8% respectively in groups E30 to E120. (2)H_2O_2 contents of skeletal muscle were clearly increased in group s E30 to E120. (3)ATP synthesis activity elevated at 30 min of exercise and returned to the baseline thereafter,whereas respiratory control index without remarkable change in groups E30 to E120. Conclusion Muscular mirol mRNA expression significantly increased during 120min of exercise as compared with the resting status and thus facilitated the mitochondrial respiratory and ATP synthesis for matching the energy demand during exercise.

OBJECTIVETo explore the impact of plasma homocysteinemia(Hcy) on contrast-induced nephropathy (CIN) after percutaneous coronary intervention (PCI) in acute coronary syndrome (ACS) patients.

METHODSConsecutive 684 ACS patients undergoing first PCI in our department between January 2013 and December 2014 were prospectively enrolled.Patients were divided into 2 groups according to the pre-procedural plasma Hcy level: high-Hcy group (Hcy≥10 μmol/L, n=404) and control group (Hcy<10 μmol/L, n=280). The CIN was defined as serum creatinine ≥ 44.2 μmol/L or 25% increase compared to baseline within 48-72 h after PCI.The baseline clinical data and the ratio of CIN were compared between the 2 groups.Multivariate logistic regression analysis was used to define the independent risk factors for CIN.

RESULTSCIN occurred in 133(19.4%) out of 684 enrolled patients, and the incidence of CIN was significantly higher in high Hcy group than in the control group (22.0%(89/404)vs. 15.7%(44/280), P=0.040). After adjusting the confounding factors, including age, acute myocardial infarction, co-morbidities(hypertension, diabetes mellitus, and old myocardial infarction), laboratory examination (level of cystatin C and uric acid), glomerular filtration rate, left ventricular ejection fraction, angiographic and procedural characteristics (3 diseased vessels, multiple stent implantation), treatment at admission (spironolactone, digoxin), multivariate logistic regression analysis showed that high Hcy was independently associated with the development of CIN (OR=1.70, 95%CI 1.60-2.64, P=0.021).

CONCLUSIONElevated Hcy prior PCI is an independent risk factor of CIN in ACS patients undergoing first PCI.

Acute Coronary Syndrome ; Diabetes Mellitus ; Glomerular Filtration Rate ; Humans ; Hyperhomocysteinemia ; Incidence ; Kidney Diseases ; Myocardial Infarction ; Percutaneous Coronary Intervention ; Risk Factors ; Ventricular Function, Left

OBJECTIVETo investigate the characteristics of the abnormalities of chromosome 17 in myeloid malignancies with complex chromosomal abnormalities (CCAs).

METHODSAbnormalities of chromosome 17 were analyzed in 73 patients with myeloid malignancies with CCAs showed by R banding and conventional karyotyping, including 21 acute myeloid leukemia (AML), 36 chronic myeloid leukemia (CML) and 16 myelodysplastic syndrome (MDS). All CCAs were further analyzed by multiplex fluorescence in situ hybridization (M-FISH).

RESULTSAmong the 73 myeloid malignancies with CCAs, chromosome 17 was the most frequently involved chromosome. It was found in 46.5% (34/73) of all cases, including 12 AML, 13 CML in blast crisis (BC) and 9 MDS. However, it was not found in the 9 CML cases in chronic phase (CP). The majority of changes were structural rearrangements which were identified in 43.8%(32/73)of all cases, among them the frequency was 52.4% (11/21), 33.3% (12/36) and 56.3% (9/16) in AML, CML and MDS, respectively. Numerical abnormalities were detected in 15.1% (11/73) cases, all were monosomy 17, and the frequency was 25.0% (3/12), 38.5% (5/13) and 33.3% (3/9) in AML, CML and MDS, respectively. Both numerical and structural abnormalities of chromosome 17 were found in 9 cases. Unbalanced translocations involving chromosome 17 were much more frequent than balanced ones. In the 3 groups, 16, 15 and 8 unbalanced translocations were found respectively. Only two kind of balanced translocations including t(15;17) in AML and t(15;17;22) in CML were found. All chromosomes were involved except chromosomes 5, 6 and 22 as partner chromosomes, the most common one was chromosome 15 (8.2%), followed by chromosome 2 (5.4%). Five of the 6 cases with translocation of chromosomes 15 and 17 were acute promyelocytic leukemia, the other case was CML-BC.

CONCLUSIONAbnormalities of chromosome 17 were the most frequently involved chromosomal aberrations in myeloid malignancies, and structural rearrangements were more common. All the numerical abnormalities were monosomy 17, unbalanced translocations were much more frequent than balanced ones.

Adolescent ; Adult ; Aged ; Child ; Chromosome Aberrations ; Chromosomes, Human, Pair 17 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics

Objective:To investigate the clinical significance of cc-chemokine receptor 7 (CCR7) as a potential diagnostic or differential marker for chronic lymphocytic leukemia (CLL).Methods:A total number of 643 patients with B-cell chronic lymphoproliferative diseases (B-CLPD) admitted to the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2018 were enrolled. The patients included 327 cases of CLL, 58 cases of mantle cell lymphoma (MCL), 34 cases of follicular lymphoma (FL), 36 cases of marginal zone lymphoma (MZL), 10 cases of hair-cell leukemia or its variants (HCL/HCLV-v), 40 cases of Waldorf′s macroglobulinemia (WM), 48 cases of CD5 +B-cell chronic lymphoproliferative disease unclassified (B-CLPD-U) and 90 cases of CD5 -B-CLPD-U. At the same time, 20 samples from healthy people from the medical examination center of our hospital were used as normal controls. Flow cytometry was used to detect the immune-phenotype and CCR7 expression level in B-CLPD patients, and Fluorescence in situ hybridization (FISH) was used to analyze the genomic alterations: the ataxia telangiectasia mutant gene (ATM) deletion, the 13q14 deletion, the P53 deletion and trisomy 12. Sanger sequencing was used to analyze gene mutations of splicing factor 3B subunit 1 (SF3B1), NOTCH1, tumor protein 53 (TP53) and immunoglobulin heavy chain variable region (IGHV). Measurement data were compared by Mann-Whitney test, and the positive rates were compared by chi-square test. The diagnostic value and optimal positive cutoff value of CCR7 were calculated using receiver operating characteristic (ROC) curve. Results:The positive rates of CCR7 expression in typical CLL and atypical CLL were 90.8% (257/283) and 84.1% (37/44), respectively, and there was no significant difference of the positive rates (χ 2=1.228, P=0.268) between groups. The positive expression rates of CCR7 in CLL, MCL, CD5 +B-CLPD-U, CD5 -B-CLPD-U, FL, WM, HCL/HCL-v and MZL were 89.9% (294/327), 10.3% (6/58), 6.3% (3/48), 8.9% (8/90), 0, 0, 0 and 13.9% (5/36) respectively, and the median mean fluorescence intensity (MFI) was 278 (246, 307), 114 (106, 128), 112 (106, 117), 110 (104, 121), 108 (105, 119), 111 (105, 124), 112 (108, 115) and 109 (105, 120) respectively. Compared with CLL, the positive expression rates of CCR7 in other types of B-CLPDs were lower significantly (χ 2=181.3, 177.8, 232, 164.7, 180.8, 62.6, 129, P<0.01). In addition, the sensitivity, specificity and accuracy of CCR7 for distinguishing CLL from other types of B-CLPD were 89.9%, 93.0% and 92.3%, respectively. The positive expression rate of CD49d in CCR7 +CLL patients was 13.9%, which was significantly lower than that in CCR7 -CLL patients (42.1%) (χ 2=7.6, P=0.01). The coincidence rate of 13q14 deletion was 50.3% in CCR7 +CLL patients, which was significantly higher than that in CCR7 -CLL patients (20%) (χ 2=6.56, P=0.01). Conclusions:The CC-chemokine receptor 7 (CCR7) antigen is an effective marker for the diagnosis and identification of chronic lymphocytic leukemia (CLL). The expression level of CCR7 in clinical specimens can distinguish CLL from other pathological subtypes of B-CLPDs.

Objective:To analyze the distribution of different SF3B1 genotypes in patients with myelodysplastic syndromes (MDS) and its prognostic value.Methods:Totally, 377MDS patients who were initially diagnosed in the First Affiliated Hospital of Nanjing Medical University from January 2014 to January 2022 were included in the retrospective analysis.The patients were divided into three different groups according to mutation stcote of SF3B1, including 317 patients with SF3B1 wild type (SF3B1 WT) (214 males and 103 females, 63(49, 71) years old),39 patients with SF3B1 K700E mutation(SF3B1 K700E(17 males and 22 females, 65(52, 73)years old)) and 21 patients with SF3B1 non-K700E mutation(SF3B1 non-K700E)(13 males and 8 females, 67(63, 73) years old). MDS-related 20 gene mutations were detected using targeted sequencing technology; Survival curves were constructed by the Kaplan-Meier method; Cox proportional hazards model was established to evaluate different factors at diagnosis on survival by univariate and multivariate analyses.. Results:Compared with SF3B1 non-K700E patients, SF3B1 K700E patients had a higher median absolute neutrophil count ( P=0.002) and were likely to be in the low/int-1 International Prognostic Scoring System (IPSS) categories ( P=0.023). A 20-gene targeted sequencing analysis showed that, compared with SF3B1 WT patients, SF3B1 K700E patients were associated with lower frequency of ASXL1 and U2AF1 mutations ( P=0.018 and P=0.003); while compared with SF3B1 non-K700E patients, the frequency of ASXL1 mutation was significantly lower in SF3B1 K700E cases ( P=0.029). Patients with SF3B1 K700E had better overall survival (OS) in comparison with SF3B1 WT and SF3B1 non-K700E in MDS patients ( P<0.001 and P=0.045, respectively). In comparison with SF3B1 WT patients, SF3B1 MUT patients had more favorable OS and progression-free survival (PFS) in MDS without excess blasts ( P<0.001 and P<0.001, respectively), but no significant difference was found in MDS with excess blasts ( P>0.05). Compared with SF3B1 WT patients, SF3B1 K700E patients had superior OS and PFS in the int-1 IPSS category ( P=0.010 and P=0.013, respectively). By multivariable analysis, the presence of SF3B1 K700Ewas an independent predictor of superior OS ( HR=0.461,95% CI 0.262-0.811, P=0.007). Conclusion:SF3B1 K700E and SF3B1 non-K700E patients had significantly improved OS in comparison with SF3B1 WT MDS patients. Furthermore, SF3B1 K700E patients were associated with a better OS compared with SF3B1 non-K700E MDS patients. SF3B1 mutation could not overcome the poor prognostic effect of excess blasts, which highlights the importance of the SF3B1 mutation subtype in risk assessment of MDS without excess blasts.

OBJECTIVESince the incidence rates and risk factor for type 2 diabetes in Chinese populations had not been well known, the aim of this study was to evaluate the impact of weight change and other risk factors on incident type 2 diabetes in Qingdao, China.

METHODSA prospective population-based cohort study was carried out, based on subjects aged 35-74 years who participated in the 'Qingdao Diabetes Survey' in 2006. Subjects were free of diabetes at baseline. A total of 1 294 subjects attended the follow up survey between 2009 and 2011. The diagnostic criteria for Diabetes was classified according to both the World Health Organization and the International Diabetes Federation 2006. A logistic regression was built using the backward stepwise selection to assess the effects of risk factors on the incident type 2 diabetes.

RESULTSDuring a 4-year follow up period, 120 cases with incident type 2 diabetes were identified, with cumulative incidence of diabetes as 11.8% . Participants who developed type 2 diabetes were significantly older, having significantly higher age-adjusted BMI/waist circumference/systolic blood pressure and total cholesterol, than those subjects who remained non-diabetic both in urban and rural areas. Among individuals with no diabetes at the baseline, factors as age, living in the rural areas, baseline BMI and weight change had all independently contributed to the development of diabetes. The multivariate adjusted relative risks (95%CIs) related to the incidence of diabetes were 1.45 (1.13-1.87), 1.93 (1.12-3.34), 1.46 (1.05-2.03) and 1.49 (1.18-1.88), respectively, for a one standard deviation increase in continuous variables. Compared with the reference group of non-obese and with stable weight, factor as weight loss >5% and BMI <28 kg/m² were independently associated with a 67% (RR = 0.33, 95% CI: 0.11-0.97)reduction in the risk of type 2 diabetes, while BMI >28 kg/m² could increase the risk across the levels of weight change. Similar trends were observed in higher waist and weight gain at baseline.

CONCLUSIONThis study confirmed the critical importance of obesity in the development of type 2 diabetes. Baseline BMI and weight gain appeared independent predictors on type 2 diabetes.

Adult ; Aged ; Body Mass Index ; Body Weight ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; Female ; Follow-Up Studies ; Humans ; Incidence ; Logistic Models ; Male ; Middle Aged ; Prospective Studies ; Risk Factors ; Weight Gain