BACKGROUND:At present,there are two main methods of isolating and purifying bone marrow mesenchymal stem cells(BMSCs):density gradient centrifugation and the whole bone marrow adherent method.The former has complicated procedures,and the latter has simple operation,but the purified outcomes are not ideal.OBJECTIVE:To establish a rat BMSCs isolated and cultured in vitro purification methods on the basis of the whole bone marrowadherence and isolation of BMSCs in combination of differential passage digestion method.METHODS:By whole bone marrow adherent culturing to isolate and differential digestion and passage of rat BMSCs,the speed ofMSCs in the process of digestion and passage was quicker than other bone marrow cells,as well as the characteristics of adherentspeed,instead of density gradient centrifugation to separate and purify MSCs,and their morphological characteristics wereobserved.Cell growth and proliferation of two kinds of culture method were compared with the density gradient centdfugationseparation.Alkaline phosphatase and oil red staining results were observed to verify BMSC differentiation capacity,to detect thecall surface markers,to validate immune properties and to test its purity.RESULTS AND CONCLUSION:The whole bone marrow adherent culture method isolated and differentially subcultured ratBMSCs.Flow cytometry and osteogenic and adipogenic culture results displayed cytoirnmunity property,purity and differentialcapacity,which have no significant difference compared with density gradient centrifugation.However,cell viability andreproductive activity are obviously elevated.

Objective To observe the changes of the heart rate turbulence(HRT)in patients with isolated systolic hypertension(ISH).Methods 85 patients with ISH,80 patients with common hypertension and 58 control subjects were investigated.Turbulence onset(TO)and turbulence slope(TS)were recorded of 24 hours dynamic electrocardiogram.Results The difference of HRT in the three groups was significant(P＜0.01).The values of TO and TS was significantly higher in common hypertension group and ISH group than in control group(P＜0.05);TS was significantly lower in ISH group than in common hypertension group(P＜0.05),but the difference of TO between ISH group with common hypertension group was not obvious.Conclusion The HRT in patients with primary hypertension is significantly lower,which is more significant in patients with ISH than in patients with common hypertension.It suggests that the cardiac autonomic nerves dysfunction perhaps has an important influence on development of hypertension and its prognosis.

Objective To investigate the efficacy and safety of levosimendan on patients with acutely decompensated heart failure (ADHF).Methods A prospective randomized and controlled study was carried out from June 2013 to June 2014.Patients were randomly divided into levosimendan group and dobutamine group,with 60 patients in each group.All patients received an intravenous infusion of levosimendan or dobutamine for 24 hours.Brain natriuretic peptide (BNP),stroke volume (SV) and left ventricular ejection fraction (LVEF) were measured.The cardiovascular mortality,rehospitalization rate,the composite endpoint differences and the incidence of adverse events were compared between two groups in 1,3,6 months after treatment.Comparisons between two groups were made using Student t-test or one-way ANOVA.Statistical analysis was performed using SPSS 17.0 software and a P value of ＜ 0.05 was considered statistically significant.Results There was no significant difference in the basic characteristics between two groups.Compared with baseline level,the plasma BNP levels,SV and LVEF were improved at 24 h in both groups (P ＜ 0.05).The cardiac function indexes were better in levosimendan group than in dobutamine group at 24 h [BNP (1147±407) pg/mL vs.(1 502±501) pg/mL,SV (60.9±9.6) mL vs.(57.3±10.3) mL,LVEF (31.6±6.0)％ vs.(28.8±5.1)％,P＜0.05].One month later,the cardiac function indexes were still better in levosimendan group than baseline [BNP (796 ± 296) pg/mL vs.(1 951 ±692) pg/mL,SV (64.6±9.5) mL vs.(52.2±9.1) mL,LVEF (33.4 ±5.8)％ vs.(25.7 ± 6.1) ％,P ＜ 0.05].After l months of treatment,the composite endpoint in levosimendan group was significantly lower than dobutamine group (5％ vs.16.3％,P =0.043).There was a downward trend of mortality and rehospitalization rate in levosimendan group in six months follow-up (P ＞ 0.05).The incidence of side effects was no statistically significant between groups (both were 13.3％).Conclusions Levosimendan is superior to that of dobutamine in improving the hemodynamic status and prognosis in ADHF patients,and the adverse reaction of levosimendan is less.

Objective To explore the effect of adipose-derived stem cells (ADSCs) on inflammatory factors in rats with lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the possible mechanism of anti-inflammatory. Methods Seventy male Sprague-Dawley (SD) rats were randomly divided into normal control group (n = 10), LPS model group (n = 30), and ADSCs intervention group (n = 30) by random number table. ALI model was reproduced by intraperitoneal injection of 8 mg/kg LPS, and the rats in ADSCs intervention group received tail vein injection of 300 μL ADSCs 30 minutes after the model reproduction, the samples of normal control group were harvested immediately without any intervention, and the specimens in remained two groups were taken at 6, 24, 72 hours respectively. Arterial partial pressure of oxygen (PaO2) and lactate level in femoral artery were determined. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum myeloperoxidase (MPO) and interleukin-10 (IL-10) in the blood of left ventricle. Lung wet/dry weight (W/D) ratio was detected by thoracotomy, and the pathological changes of lung tissue were observed under an optical microscope. Western Blot was used to detect the protein expression of nuclear factor-κB (NF-κB) in lung tissue of rats. Results Compared with the normal control group, the damage degree of lung tissue of LPS model group was significantly heavier from 6 hours, and lung W/D ratio, blood lactate, MPO, IL-10 and expression level of NF-κB in lung tissue were significantly increased respectively, while PaO2 was decreased significantly. Compared with LPS model group, the damage degree of lung tissue of ADSCs intervention group was significantly reduced from 6 hours, and lung W/D ratio, blood lactate, MPO, and NF-κB expression in lung tissue were significantly decreased, while PaO2 was increased significantly, and it became normal at 72 hours [lung W/D ratio: 5.33±0.29 vs. 5.77±0.42 at 6 hours, 5.14±0.46 vs. 5.43±0.38 at 72 hours; blood lactate (mmol/L): 3.6±1.0 vs. 5.7±1.1 at 6 hours, 3.1±1.0 vs. 3.8±1.2 at 72 hours; blood MPO (μg/L): 1.50±0.90 vs. 2.70±1.85 at 6 hours, 0.46±0.30 vs. 0.71±0.22 at 72 hours; NF-κB (gray value): 0.40±0.11 vs. 0.50±0.09 at 6 hours, 0.24±0.03 vs. 0.33±0.06; PaO2 (mmHg, 1 mmHg = 0.133 kPa): 78.0±4.1 vs. 74.5±3.2 at 6 hours, 89.3±9.4 vs. 81.9±3.4 at 72 hours; all P < 0.05]. The IL-10 level was significantly higher than that of LPS model group only at 24 hours (ng/L: 27.75±15.49 vs. 17.52±6.56, P < 0.05). Conclusion ADSCs can effectively relieve the inflammatory response of ALI induced by LPS, probably by inhibiting the expressions of NF-κB and blocking the release of inflammatory cytokines.

Objective To investigate the relationship between serum indoxyl sulfate(IS),Clusterin and coronary artery lesions in patients with acute coronary syndrome(ACS)and its predictive value for short-term prognosis.Methods A total of 120 ACS patients who were admitted to the First Affiliated Hospital of Zhengzhou University from June 2021 to October 2022 were selected as the research subjects(ACS group),and another 80 healthy volunteers who underwent physical examinations in our hospital during the same period were selected as the control group.Serum IS and Clusterin levels of patients in the ACS group and control group were detected by enzyme-linked immunosorbent assay.ACS patients were divided into a single-vessel disease group(n=43),a double-vessel disease group(n=49)and a multi-vessel disease group(n=28)according to the number of coronary artery lesions,divided into a mild lesion group(n=37),moderate lesion group(n=48)and severe lesion group(n=35)according to the Gensini score,and divided into a good prognosis group(n=85)and a poor prognosis group(n=35)according to the short-term prognosis.The influencing factors for poor short-term prognosis in ACS patients were analyzed by univariate and multivariate logistic regression,and the predictive value of serum IS and Clusterin levels for poor short-term prognosis in ACS patients was evaluated by receiver operating characteristic(ROC)curve.Results The serum IS and Clusterin levels of patients in the ACS group were significantly higher than those in the control group(P＜0.05).The serum IS and Clusterin levels of patients in the single-vessel disease group,double-vessel disease group and multi-vessel disease group increased in turn(P＜0.05).The serum IS and Clusterin levels of patients in the mild lesion group,moderate lesion group and severe lesion group increased in turn(P＜0.05).The incidence of poor short-term prognosis in 120 ACS patients was 29.17％(35/120).Univariate analysis showed that,ACS type,Gensini score,KILLIP grade,number of coronary artery lesions,IS,and Clusterin were related to the short-term prognosis in ACS patients(P＜0.05).Multivariate logistic regression analysis showed that,increased Gensini score,increased IS,increased Clusterin,KILLIP grade≥Ⅲ,and multi-vessel coronary artery disease were independent risk factors for poor short-term prognosis in ACS patients(P＜0.05).The area under the ROC curve(AUC)for predicting poor short-term prognosis in ACS patients based on serum IS level was 0.747(95％confidence interval:0.529-0.946),with sensitivity of 71.43％,specificity of 74.12％,accuracy of 73.33％,and Youden index of 0.455.The AUC for predicting poor short-term prognosis in ACS patients based on serum Clusterin level was 0.693(95％confidence interval:0.439-0.951),with sensitivity of 68.57％,specificity of 70.59％,accuracy of 7.00％,and Youden index of 0.392.The AUC for predicting poor short-term prognosis in ACS patients based on both serum IS and Clusterin levels was 0.825(95％confidence interval:0.687-0.968),with sensitivity of 82.86％,specificity of 83.53％,accuracy of 83.33％,and Youden index of 0.664.Conclusion Increased serum IS and Clusterin levels are associated with aggravation of coronary artery disease and poor short-term prognosis in ACS patients,and a combination of the two has a high predictive value for poor short-term prognosis in ACS patients.

Objective: To explore the treatment technique, occurrence and development patterns of such radiation injuries as in a major radiological accident in which a victim suffered mild bone marrow radiation sickness combined grade degree Ⅲ acute radiation induced skin injury, based on his dose estimation, clinical manifestations and disease treatments. Methods: History inquiry in detail, earlier physical dose estimation and biological dose estimation were conducted in conjunction with analyzing the chromosome aberration of peripheral blood lymphocytes. The physical dose was estimated by Monte Carlo method.The systematic laboratory and imaging examination was performed to evaluate the condition. The comprehensive analysis was conducted to determine the diagnosis and treatment plan. Results: At 3d after the exposure, "Ren" felt mild pain and discomfortable on the skin of the right index finger. The body of the right hand index finger was covered with blister at 21 d after exposure.The estimation of biological dose was 0.43 Gy (95%CI: 0.31-0.58 Gy), and the physical dose was estimated to be 36-164 Gy for each part of the right hand finger. The hematopoietic system, immune system and endocrine system were normal. The liver function index value was transiently increased. The liver damage resulted from the use of antibiotic-induced combined with the patient′s past medical history and admission examination result, and the relevant antibiotics were discontinued. The liver function returned to normal after liver protection treatment. At 22 d after irradiation, a right finger incision and decompression surgery were performed. The stem cells were extracted and implanted into the right index finger. After 59 days of hospitalization, there was no obvious discomfort in the body, and the right index finger recovered well, as well as the pain significantly relieved, and the knuckle activity was basically normal. Conclusions The patient with excessive radiation and grade Ⅲ acute radiation skin injury was successfully treated, and local application of autologous adipose-derived stem cell transplantation achieved good result .

Objectives:To report a novel mutation site in the pathogenic gene TTR of transthyretin cardiac amyloidosis(ATTR-CA),and to identify family members at risk,and provide suitable clinical diagnosis and treatment. Methods:A retrospective analysis was conducted on the clinical data of the proband with ATTR-CA who visited the Department of Cardiology,the First Affiliated Hospital of Zhengzhou University in March 2021.The proband underwent whole exome sequencing using high-throughput methods to detect mutation genes.Sanger sequencing was used to test candidate pathogenic loci in suspected family members,and relevant literature was reviewed. Results:Among 51 individuals spanning 5 generations in the pedigree,10 family members(including the proband)carried the heterozygous TTR gene c.163A>G mutation,resulting in the amino acid residue at position 55 changing from lysine(Lys)to glutamic acid(Glu).This mutation follows an autosomal dominant inheritance pattern,with early onset in adulthood,rapid progression,and presenting as a mixed-type ATTR-CA.Five mutation carriers had different clinical manifestations,while the remaining 5 mutation carriers,who are at younger age,have not yet shown symptoms.Within the pedigree,7 individuals died(the proband's uncle[Ⅱ-1]who died from stroke at 65 years old,the rest 6 family members died from heart disease before the age of 50). Conclusions:According to the American College of Medical Genetics and Genomics guidelines,TTR gene Lys55Glu mutation is classified as likely pathogenic,this mutation site has not been reported in the literature before.Present study adds clinical evidence that might broaden the spectrum of TTR mutations.

Objective To investigate the relationship between serum indexes and the degree of liver fibrosis in chronic hepatitis B(CHB)patients with HBeAg-negative and normal ALT,and to establish a new non-invasive model for predicting liver fibrosis in CHB patients.Methods The clinical data of 679 HBeAg-negative chronic HBV infected patients with normal ALT who underwent liver biopsy from October 2012 to December 2021 were retrospectively analyzed.Among these patients,they were categorized into the control group(S1,observation group)the and significant fibrosis group(S2/S3/S4,control group)based on liver biopsy results.The LASSO regression model was used for covariates selection and the restricted cubic splines model was used to examine nonlinear associations between covariates and outcomes.We used Logistic regression models to establish predictive models.Results Liver biopsy showed that 48.7%of the patients had obvious fibrosis(S≥2).GGT shows a nonlinear relationship with the degree of liver fibrosis.AST and PT show a positive relationship with the liver fibrosis degree,respectively.The area under the ROC curve(AUC)of GGT + PT + AST is 0.68(95%CI:0.64～0.72),and this model performed better than models established using GPR,APRI,and FIB-4.Conclusion The prediction model of GGT + PT+AST has high predictive value on the severity of liver fibrosis among CHB patients whose HBeAg is negative.