Difference analysis of chemical components in roots and rhizomes of 3 medicinal plants of Gentiana from the same origin / 中国药房

OBJECTIVE To provide reference for exploring alternative resources of Gentiana rigescens from the plants of Gentiana. METHODS The contents of four components （gentiopicroside， swertiamarin， swertioside and amarogentin） in the roots and rhizomes from 3 plants of Gentiana （G. rigescens， G. cephalantha， G. delavayi） were determined by high-performance liquid chromatography （HPLC）. The chemical compositions in the above roots and rhizomes were identified by ultra-performance liquid chromatography electrospray ionization quarter-time of flight mass spectrometry （UPLC-ESI-Q-TOF-MS）， and the differences were analyzed by principal component analysis （PCA）. RESULTS Four active components such as gentiopicroside， swertiamarin， swertioside and amarogentin were detected in the roots and rhizomes of G. rigescens and G. cephalantha， and the contents of the four components were similar in both. The contents of gentiopicroside in the root and rhizome of G. cephalantha and G.rigescens were more than four times of the limit standard of the Chinese Pharmacopoeia （Part Ⅰ） in 2020； However， only swertiamarin， swertioside and amarogentin were detected in the roots and rhizomes of G.delavayi， and the contents of swertioside and amarogentin were 34.12 and 8.81 times of those of G. rigescens， respectively. In addition， a total of 33 compounds 术。E-mail：515227235@qq.com were identified from the roots and rhizomes of 3 plants of Gentiana by UPLC-ESI-Q-TOF-MS， mainly iridoids. Additionally， G. rigescens and G. cephalantha contained xantones， G. delavayi contained flavonoids. PCA showed that there was a small difference between G. rigescens and G. cephalantha； however， there was a big difference between G. delavayi and G. rigescens. CONCLUSIONS The difference between the roots and rhizomes of G. cephalantha and G. rigescens from the same origin is small and there is substitutability； while the difference in the chemical components from roots and rhizomes between G. delavayi and G. rigescens is great and G. delavayi cannot be used as medicine instead of G. rigescens.

Liriodendrininhibits cardiomyocyte apoptosis in acute myocardial infarction rats / 中华老年医学杂志

Objective:To investigate the protective effect of liriodendrin on acute myocardial infarction in rats and to explore the related mechanisms.Methods:From January to December 2019, 30 SPF male Wistar rats with a body weight of(200±10)g were randomly divided into a sham operation group, a control group, and a liriodendringroupwith 10 rats in each group using the numerical sampling method.The liriodendron group was intragastrically administered with a liriodendrinsolution(10 ml/kg)once a day from 5 days before myocardial infarction model construction to 3 days after surgery.The control group and the sham surgery group were intragastrically administered with 10 ml/kg normal saline.After surgery, high-sensitivity troponin T levels were measured in the three groups.Cardiac function of the rats was assessed using echocardiography on the 3rd day post-surgery.Then, the rats were sacrificed, followed by hematoxylin-eosin(HE)staining and TdT-mediated dUTP nick-end labeling(TUNEL)staining of cardiac tissues and measurement of interleukin(IL)-1β and tumor necrosis factor(TNF-α)levels.Western blot and real-time polymerase chain reaction(PCR)were used to detect the expression of apoptosis-related proteins and transcriptional activity.Results:High-sensitivity troponin T levels in the liriodendrin group[(1.74±0.63)μg/L]were lower than in the myocardial infarction group[(3.54±1.60)μg/L]at 2 hours after surgery( t=2.69, P<0.05). Echocardiography showed that, compared with the myocardial infarction group, the ejection fraction was higher in the liriodendrin group, and the left ventricular end-diastolic diameter, left ventricular end-systolic diameter, left ventricular end-diastolic volume and left ventricular end-systolic volume were lower in the liriodendrin group( P<0.05). Histological staining showed that the myocardial tissue of the control group was severely damaged, with infiltration of a large number of in flammatory cells.The number of TUNEL-positive cells in the liriodendrin group(56.66±2.414)was statistically significantly reduced, compared with in the myocardial infarction group(76.55±1.843)( t=6.55, P<0.05). The levels of IL-1β and TNF-α in the myocardial infarction group were higher than those in the liriodendrin group( P<0.05). The expression of apoptosis-related proteins in the liriodendrin group was lower( P<0.05)and the transcriptional activity of mRNA was also lower( P<0.05)than in the myocardial infarction group. Conclusions:Liriodendrin may protect cardiomyocytes after myocardial infarction in rats by inhibiting local inflammation and cell apoptosis.

Effects of overexpression of human vascular endothelial growth factor 165 and human tissue inhibitor of metalloproteinase 1 on rats with myocardial infarction / 中华急诊医学杂志

Objective:To investigate the effects of recombinant adenovirus with human vascular endothelial growth factor 165 (Ad-hVEGF 165) and recombinant adenovirus with human tissue inhibitor of metalloproteinase 1 (Ad-hTIMP-1) on rats with myocardial infarction (MI) and its mechanism. Methods:A total of 30 healthy 8-week-old male Wistar rats were randomly divided into 5 groups: sham-operated group (sham), virus control group (Ad-Track), Ad-hVEGF 165 group, Ad-hTIMP-1 group and Ad-hVEGF 165+Ad-hTIMP-1 group (hVEGF 165+hTIMP-1) ( n=6 per group). Except the sham group, all rats were ligated the left anterior descending coronary artery to induce MI model with ST-segment elevation and Q waves or T-wave inversion on electrocardiogram and local myocardial whitening. The corresponding recombinant adenovirus comprising 100 μL (1×10 10 VP/100 μL) combined with NaCl solution was injected into the myocardial infarction area at four points respectively. The sham group received no treatment. After 4 weeks, all rats were sacrificed after echocardiography was completed and heart tissues were collected. The expression of hVEGF 165 and hTIMP-1 were detected by immunohistochemistry. The mRNA expression of apoptosis-related factors were detected by real-time PCR. The protein expression of apoptosis-related factors were detected by immunohistochemistry. Differences between groups were determined by One-way analysis of variance. Multiple comparisons between groups were performed using the least significant difference t-test. Results:(1) Both heart rate (HR) (480.83±24.09) beats/min, left ventricular end-diastolic dimension (LVEDD) (6.88±0.44) mm and left ventricular end-systolic dimension (LVESD) (4.85±0.42) mm were increased in the Ad-Track group than those in the sham group (433.16±17.86) beats/min, (6.20±0.45) mm, (4.06±0.70) mm (all P<0.05), and left ventricular ejection fraction (LVEF) (62.70±3.17) % and left ventricular fractional shortening (LVFS) (29.52±1.88) % were significantly decreased in the Ad-Track group than those in the sham group (72.78±5.44)%, (29.52±1.88) % (both P<0.01). Compared with the Ad-Track group, LVEF (71.50±6.23) % and LVFS (36.17±5.27) % in the hVEGF 165-hTIMP-1 group were significantly increased (both P<0.01), and LVEDD (6.22±0.39) mm and LVESD (4.13±0.23) mm were decreased (both P<0.05). LVEF and LVFS in the hVEGF 165-hTIMP-1 group were increased significantly than those in the Ad-hVEGF 165 group (64.65±4.00) %, (30.95±2.57) % (both P<0.05). The mRNA expression of BCL2-associated X protein (Bax), cysteine aspartate specific proteinase 3 (Caspase-3) and BCL-xL/BCL-2-associated death promoter (Bad) in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-Track group ( P<0.01 or P<0.05), and B-cell lymphoma/leukemia-2 (Bcl-2) in the hVEGF 165-hTIMP-1 group were increased than those in the Ad-Track group ( P<0.01). The mRNA expression levels of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-hVEGF 165 group (both P<0.05). There was no statistically difference in the mRNA expression of Bax, Caspase-3, Bad, and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). The protein expression of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were significantly decreased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.01), and the protein expression of Bcl-2 in the hVEGF 165-hTIMP-1 group was increased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.05). There were no statistically differences in the protein expression of Bax, Caspase-3 and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). Conclusions:Ad-hVEGF 165 and Ad-hTIMP-1 can improve cardiac contractile function of MI rats and the beneficial effects are largely attributable to inhibiting myocyte apoptosis. The combination of hVEGF 165 and hTIMP-1 may have a synergistic effect on MI.

Application of Biomarkers for the Prediction and Diagnosis of Bone Metastasis in Breast Cancer / 한국유방암학회지

The most common metastatic site of breast cancer is the bone. Metastatic bone disease can alter the integrity of the bone and cause serious complications, thereby greatly reducing health-related quality of life and leading to high medical costs. Although diagnostic methods and treatments for bone metastases (BM) are improving, some patients with early breast cancer who are at high risk of BM are not diagnosed early enough, leading to delayed intervention. Moreover, whole-body scintigraphy cannot easily distinguish BM from nonmalignant bone diseases. To circumvent these issues, specific gene and protein biomarkers are being investigated for their potential to predict, diagnose, and evaluate breast cancer prognosis. In this review, we summarized the current biomarkers associated with BM in breast cancer and their role in clinical applications to assist in the diagnosis and treatment of BM in the future.

Application of Biomarkers for the Prediction and Diagnosis of Bone Metastasis in Breast Cancer / 한국유방암학회지

The most common metastatic site of breast cancer is the bone. Metastatic bone disease can alter the integrity of the bone and cause serious complications, thereby greatly reducing health-related quality of life and leading to high medical costs. Although diagnostic methods and treatments for bone metastases (BM) are improving, some patients with early breast cancer who are at high risk of BM are not diagnosed early enough, leading to delayed intervention. Moreover, whole-body scintigraphy cannot easily distinguish BM from nonmalignant bone diseases. To circumvent these issues, specific gene and protein biomarkers are being investigated for their potential to predict, diagnose, and evaluate breast cancer prognosis. In this review, we summarized the current biomarkers associated with BM in breast cancer and their role in clinical applications to assist in the diagnosis and treatment of BM in the future.

Effect of noninvasive ventilation on heart failure after acute myocardial infarction in rats and the underlying mechanisms / 中华老年医学杂志

Objective To study the effect of noninvasive ventilation on heart failure(HF)in rats after acute myocardial infarction(AMI)and the mechanisms involved.Methods A total of 30 Wistar rats were randomly divided into three groups:the noninvasive ventilation group,the non-treatment group and the sham operation group(n=10 in each group).The model for HF after AMI was established by left anterior descending coronary artery ligation.Serum levels of B-type natriuretic peptide-45 (BNP-45),70 000 heat-shock protein(HSP70),matrix metalloproteinases(MMP-2 and-9),tumor necrosis factor-α(TNF-α)and echocardiography parameters,such as left atrium(LA)diameter,left ventricular(LV)end-diastolic diameter,interventricular septal(IVS)thickness,and left ventricular ejection fraction(LVEF),were measured at 3,5,7 and 14 days after operation in each group.Results Compared with the non-treatment group,LVEDD was reduced and LVEF was increased in the noninvasive ventilation group at 14 days after operation(155.92±14.74)mm/m2 vs.(149.35±11.29)mm/m2,(92.13±3.72)％ vs.(76.39±9.24)％,(P＜0.05),while LA diameter and IVS thickness showed no significant difference(P＞0.05).Compared with the non-treatment group,serum levels of BNP45,MMP-2,MMP-9,and TNF-α were decreased and HSP70 levels were increased in the noninvasive ventilation group at 3,5,7,and 14 days after operation (P ＜ 0.05).In the noninvasive ventilation group,serum levels of BNP45,MMP-2,MMP-9 and TNF-α at 5,7,and 14 days after operation were lower than those at 3 days after operation,and serum HSP70 levels at 7 and 14 days after operation were higher than those at 3 days after operation (P ＜ 0.01).Conclusions Noninvasive ventilation can effectively reduce the inflammatory response in rats with HF after AMI and reduce myocardial ischemia-reperfusion injury.It is an effective treatment for HF after AMI.

Investigation of relationship between mean platelet volume and saphenous vein graft restenosis after coronary artery bypass grafting / 中华老年医学杂志

Objective To investigate the relationship between mean platelet volume(MPV)and saphenous vein graft restenosis in patients receiving coronary artery bypass grafting(CABG),and to analyze the clinical significance of MPV in the prediction of restenosis after CABG.Methods A total of 354 patients admitted into Tianjin chest hospital from September 2009 to September 2014 with suspected myocardial ischemic events 3 to 5 years after CABG treatment was enrolled for a retrospective analysis.According to the coronary angiography(CAG)results,patients were divided into the vein bridge vascular lesion group(saphenous vein graft diseases,SVGD)(n=233)and the venous bridge vascular patency group(saphenous vein graft,SVG)(n=121).Paired t test was used to analyze the relationship between different factors and the bridge vascular patency.The binary logistic regression was used to analyze the effects of MPV and other factors on bridge vascular patency.Venous bridge stenosis ＞ 50％ was considered to be clinically significant and to damage myocardial blood supply.Results The MPV was higher in the SVGD group than the SVG group [(10.2±1.5)fl vs.(9.6±1.5)fl,P＜0.01].The logistic regression analysis showed that MPV(OR =1.268,95％CI:1.053-1.570,P=0.014),age(OR =1.007,95％CI:1.038-1.117,P=0.000),gender (OR=0.452,95％CI:0.250-0.816,P=0.008),diabetes mellitus(OR=2.319,95％CI:1.221-4.405,P =0.010)were the independent risk factors for venous bridge stenosis in the two groups,gender(OR=0.495,95％CI:0.251-0.976,P=0.042),diabetes mellitus(OR =2.237,95％CI:1.105-4.527,P =0.025),MPV(OR=1.334,95％CI;1.050 1.694,P=0.018),fibrinogen(OR=1.654,95％CI:1.020-2.682,P =0.041)were the independent risk factors for venous bridge stenosis in non-elderly patients,and age(OR =1.178,95％CI:1.116-1.244,P =1.178)was an independent risk factor for vein graft stenosis in elderly patients.The restenosis rate was higher in patients with MPV ≥ 12 fl(92.6％ or 25/27) than in the patients with MPV ＜ 12 fl(63.6％ or 208/327).The receiver operating characteristics(ROC) curve showed that the areas under the curve of MPV,age,gender,diabetes,fibrinogen were 0.610,0.657,0.394,0.626,0.654,respectively,and the area under the curve of joint diagnosis was 0.796,showing that joint prediction value was higher than any single prediction value(P＜0.01).Conclusions MPV level is an independent risk factor for vein graft stenosis,and has higher predictive value in combination with age,gender,diabetes and fibrinogen.

Relationship between serum levels of osteoproteins, inflammatory cytokines and coronary heart disease and disease severity / 中华危重病急救医学

Objective To explore the relationship between serum levels of osteoprotein (OPG), soluble nuclear factor-κB receptor activator ligand (sRANKL), inflammatory factors and coronary heart disease (CHD) and its severity. Methods The patients who underwent coronary angiography (CAG) due to chest pain admitted to department of cardiology of Tianjin Chest Hospital from April 2017 to December 2018 were enrolled, and they were divided into CHD group and non-CHD group according to the CAG results. The gender, age, history of hypertension, smoking history, diabetes, the levels of cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein AI (apoAI), apolipoprotein B (apoB), lipoprotein (a) [Lp (a)], MB isoenzyme of creatine kinase (CK-MB) and other clinical data of patients were collected. The serum levels of OPG, sRANKL, matrix metalloproteinase-9 (MMP-9), monocyte chemotactic protein-1 (MCP-1), insulin-like growth factor-1 (IGF-1) and interleukin-6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). According to the results of CAG, the patients with CHD were divided into single-, double-, triple-branch coronary artery lesion groups, and the relationship between the levels of serum OPG, sRANKL, inflammatory factors and the degree of coronary artery lesions was observed. Multivariate Logistic regression was used to analyze the risk factors of CHD, and receiver operating characteristic (ROC) curve was plotted to analyze the predictive value of main risk factors for CHD. Results A total of 472 patients were enrolled in the final analysis during the study period, including 264 patients in the CHD group, 208 patients in the non-CHD group, 79 patients in the CHD group with single-branch disease, 75 patients with double-branch disease, and 110 patients with three-branch disease. ① Compared with the non-CHD group, the CHD group had more older male patients, as well as higher proportion of hypertension and diabetes, the levels of serum Lp (a) and CK-MB were significantly increased, and the levels of serum HDL-C and apoAI were significantly lowered. There was no statistically significant difference in serum TC, LDL-C, or apoB between the two groups. The levels of serum OPG, MMP-9, MCP-1, IGF-1 and IL-6 in the CHD group were significantly higher than those in the non-CHD group [OPG (μg/L): 1.79±0.50 vs. 1.50±0.30, MMP-9 (μg/L): 57.91 (33.50, 130.46) vs. 38.33 (29.43, 109.78), MCP-1 (μg/L):298.30 (207.96, 537.16) vs. 252.73 (165.22, 476.01), IGF-1 (μg/L): 734.03±486.11 vs. 217.75±126.45, IL-6 (ng/L):64.76±40.25 vs. 48.60±15.80, all P < 0.05], and the levels of serum sRANKL was significantly lower than that in the non-CHD group (ng/L: 344.31±122.14 vs. 378.74±109.27, P < 0.05). ② The serum OPG level showed a slight upward tendency with the increase in the number of coronary artery lesions, and the sRANKL level showed a slight downward tendency [OPG (μg/L) in the single-, double-, triple-branch coronary artery lesion groups was 1.74±0.49, 1.76±0.50, 1.85±0.52, and sRANKL (ng/L) was 354.96±116.64, 340.05±124.24, 339.57±125.03, respectively) without statistically significant differences (all P > 0.05). The levels of IGF-1 and IL-6 were increased with the number of coronary artery lesions [IGF-1 (μg/L) in the single-, double- and triple-branch coronary artery lesions groups was 372.13±258.42, 676.06±350.29, 1 033.47±468.06, and IL-6 (ng/L) was 48.87±16.72, 65.36±18.84, 75.76±22.72, respectively], and the differences among different lesion groups were statistically significant (all P < 0.01). Correlation analysis showed that IGF-1 level was significantly positively correlated with the number of coronary artery lesions (r = 0.612, P < 0.01), while IL-6 was not correlated with the number of coronary artery lesions (r = 0.185, P > 0.05).③ Multivariate Logistic regression analysis showed that elevated serum OPG and IGF-1 levels were risk factors for CHD [OPG: odds ratio (OR) = 1.995, 95% confidence interval (95%CI) = 1.936-2.067, P = 0.012; IGF-1: OR = 1.009, 95%CI = 1.004-1.015, P = 0.001]. ④ ROC curve analysis showed that the area under ROC curve (AUC) of OPG and IGF-1 was 0.716 and 0.867, respectively. When the cut-off value of OPG was 1.13 μg/L, the sensitivity was 81.7%, the specificity was 58.1%; when the cut-off value of sRANKL was 401.20 μg/L, the sensitivity was 69.7%, the specificity was 95.7%. Conclusions CHD was associated with increased in OPG, related inflammatory cytokines including MMP-9, MCP-1, IGF-1 and IL-6, and decreased in sRANKL. The level of IGF-1 was positively correlated with the severity of CHD. The serum levels of OPG and IGF-1 were risk factors for CHD, which had good predictive value for CHD.

Polymorphisms of T149C and T950C genes in osteoprotegerin promoter sites and the relationship between serum osteoprotegerin levels and coronary heart disease / 中华危重病急救医学

Objective To explore the polymorphisms of T149C and T950C gene in osteoprotectin (OPG) promoter sites and the levels of serum OPG and soluble nuclear factor-κB receptor activator ligand (sRANKL) and the incidence of coronary heart disease (CHD). Methods 528 patients in Tianjin suspected of CHD and underwent coronary angiography (CAG) who admitted to the department of cardiology of Tianjin Chest Hospital from April 2017 to December 2018 were enrolled. According to the CAG results, they were divided into two groups: CHD group (n = 302) and non-CHD group (n = 226). The gender, age, history of hypertension, family history of CHD, diabetes, levels of blood lipid parameters in serum and other clinical data of patients were recorded. The levels of serum OPG and sRANKL were measured by enzyme-linked immunosorbent assay (ELISA). T149C and T950C gene polymorphisms were analyzed by polymerase chain reaction-restriction endonuclease fragment length polymorphism (PCR-RFLP) methods. Hardy-Weinberg genetic balance test was performed for alleles. Binomial classification multivariate non-conditional Logistic regression method was used to analyze the relationship between T149C and T950C gene polymorphisms, serum levels of OPG and sRANKL and CHD. Results All patients were enrolled in the final analysis. The serum level of OPG in CHD group was significantly higher than that in non-CHD group (μg/L: 1.76±0.49 vs. 1.47±0.29, P < 0.01), the serum level of sRANKL was significantly lower than that in non-CHD group (ng/L: 342.14±121.38 vs. 376.63±108.66, P < 0.05). Logistic regression analysis showed that after adjusting for age, gender, blood lipid parameters, diabetes and other factors, the increase in serum OPG level was an independent risk factor for CHD [odds ratio (OR) = 1.995, 95% confidence interval (95%CI) = 1.935-2.066, P = 0.012]. PCR-RFLP results showed that TT, TC and CC genotypes were found in T149C and T950C of OPG promoter. According to Hardy-Weinberg equilibrium test, the polymorphisms of OPG T149C and T950C accorded with Hardy-Weinberg law, achieving genetic balance with representative of the population. The frequencies of TT, TC, CC and alleles T and C in T149C genotypes of non-CHD group were 53.5%, 42.9%, 3.6%, 75.0% and 25.0%, respectively, and they were 43.1%, 50.3%, 6.6%, 68.2% and 31.8%, respectively in CHD group. There were statistically significant differences in genotype and allele frequencies between the two groups (all P < 0.05). It was shown by Logistic regression analysis that the risk of CHD in TC+CC genotype of T149C was 1.86 of TT genotype (OR = 1.86, 95%CI = 1.24-2.78, P = 0.003). It was suggested that C allele might be a susceptible gene for CHD. In non-CHD group, the frequencies of TT, TC, CC, and alleles T and C in T950C genotypes were 39.8%, 46.5%, 13.7%, 63.1% and 36.9%, respectively. They were 39.4%, 43.4%, 17.2%, 61.1% and 38.9%, respectively in CHD group. There were no significant differences in genotype and allele frequencies between the two groups (all P > 0.05). Logistic regression analysis showed that TC+CC genotype of T950C was not related with CHD. Conclusions The increased level of serum OPG was closely related with CHD and could be used as a risk factor for CHD. The cases carried OPG T149C TC+CC genotype might have the risk suffering CHD. C allele is might be a susceptible gene.

Establishment and evaluation of mouse models of septic myocardial injury / 中华危重病急救医学

Objective Mouse models of sepsis-induced myocardial injury by intraperitoneal injection of lipopolysaccharide (LPS) was established in order to provide a reliable method for the research of pathogenesis of sepsis-induced myocardial injury. Methods According to the method of random number table, a total of 150 male C57BL/6 mice were divided into five groups: NC group, sham group, and LPS 10, 12, 15 mg/kg groups, with 30 in each group. Septic myocardial injury was induced by intraperitoneal injection LPS in mice; sham group was injected with equal 0.9% saline; while there was no treatment in mice of NC group. Fifteen of the 30 mice in each group were used to observe the general status of mice before and after LPS or saline injection. Twenty-four hours after LPS or saline injection, the left ventricular function was assessed by echocardiography, serum level of cardiac troponin (cTnI) was determined by enzyme linked immunosorbent assays (ELISA), and the cardiac histomorphology and ultrastructure were observed; the other 15 mice were used to monitor the 7-day mortality after LPS or saline injection. Results The mice challenged to LPS displayed symptoms of sepsis, such as depression, ruffled fur, and diarrhea. Compared with NC group, left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS) were significantly decreased at 24 hours after LPS administration in LPS 10, 12, 15 mg/kg groups [LVEF: 0.459±0.044, 0.432±0.034, 0.348±0.064 vs. 0.588±0.019, LVFS: (22.36±2.60)%, (20.78±1.91)%, (16.27±3.31)% vs. (30.55±1.30)%, all P < 0.01], and cTnI levels were significantly increased (ng/L: 270.40±43.50, 281.14±41.79, 298.39±42.05 vs. 192.59±16.90, all P <0.01). Myocardium injury was observed in three LPS groups, myocardial fibrosis, interstitial edema, erythrocyte leakage and infiltrating inflammatory cells were observed under light-microscope; ultrastructural changes disorderly arranged in cardiac muscle fibers, mitochondrial swelling and even partly missing mitochondria cristae were found under transmission electron microscope (TEM), and the higher of the dose, the more sever of the damage. There was no significant difference between sham group and NC group. The 7-day mortality in LPS 10, 12, 15 mg/kg groups were 33.3%, 53.3% and 86.7%, respectively, while no death in the NC group and sham group. Conclusion For establishing the mouse model of sepsis-induced myocardial injury, intraperitoneal injection with 12 mg/kg LPS is a preferable choice in our research.

Effects of overexpression of human tissue inhibitor of metalloproteinase-1 on the inflammatory response in rats with myocardial infarction and related mechanisms / 中华心血管病杂志

Objective@#To observe the effects of recombinant adenovirus with human tissue inhibitor of metalloproteinase-1(Ad-hTIMP-1) on the inflammatory response in rats with myocardial infarction (MI) and explore the related mechanisms.@*Methods@#The male Wistar rats were randomly divided into sham-operated group, saline group, Ad-Track group and Ad-hTIMP-1 group according to the random number table (n=8 each group). MI was induced by ligation of the left anterior descending coronary artery and MI rats were injected with saline, Ad-Track and Ad-hTIMP-1, respectively. Sham-operated rats received similar surgical procedure without ligation of the left anterior descending coronary artery. After 4 weeks, the cardiac function was measured by echocardiography, then rats were sacrificed and hearts were removed for morphological and biological analysis. The morphology of myocardial tissue in each group was detected by HE staining and Masson staining. The mRNA expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and C-reactive protein(CRP) were detected by real-time PCR. Immune histochemical staining was performed to observe the protein expression levels of IL-6 and CRP.@*Results@#(1) Left ventricular end systolic dimension derived from echocardiography was increased in saline group ((5.10±0.72) mm) and Ad-Track group ((4.88±0.64) mm) compared to sham-operated group ((4.25±0.46) mm), which was reduced in Ad-hTIMP-1 group ((4.13±0.35) mm, all P<0.05). The left ventricular ejection fraction was (72.46±5.74)%, (64.27±8.52)%, (64.65±3.90)%, and (71.55±6.95)%, the fractional shortening was (36.90±4.97)%, (29.03±3.40)%, (30.95±2.51)%, and (36.31±5.68)% in sham-operated group, saline group, Ad-Track group and Ad-hTIMP-1 group, respectively. The left ventricular ejection fraction and fractional shortening in saline group and Ad-Track group were lower than those in sham-operated group and Ad-hTIMP-1 group (all P<0.05). (2) Necrosis of myocardial cells was not found and a small amount of immune cell infiltration and interstitial fibrosis were observed on HE and Masson stained myocardial sections of Ad-hTIMP-1 group. (3) Real-time PCR showed that mRNA expressions of TNF-α, IL-6, IL-10 and CRP were lower in Ad-hTIMP-1 group than in saline group. mRNA expressions of TNF-α, IL-10 and CRP were lower in Ad-hTIMP-1 group than in Ad-Track group (all P<0.05). (4) Immune histochemical staining showed that protein expressions of IL-6 and CRP were higher in saline group and Ad-Track group than those in Ad-hTIMP-1 group (all P<0.05).@*Conclusion@#Recombinant adenovirus Ad-hTIMP-1 can improve cardiac function in rats with myocardial infarction via inhibiting the inflammatory response and downregulating the expression of TNF-α, IL-6 and CRP.

Protective effects of MicroRNA-214 on myocardial injury induced by myocardial ischemia and reperfusion and the regulation mechanism of its downstream protein kinase / 中华老年医学杂志

Objective To investigate the protective effects of MicroRNA-214 on myocardial injury induced by myocardial ischemia and reperfusion,as well as the regulation mechanism of PI3K and its downstream protein kinase B(AKT)and FoxO1(PI3K / AKT / FoxO1).Methods Wistar rats were randomly divided into 4 groups:sham operation group(Sham group),myocardial ischemia reperfusion injury(IRI)group(IRI group),microRNA-214+sham operation group(MS group),microRNA-214+IRI group(MI group),(n=10,each).The cardiac function was detected at 6 h after ischemia-reperfusion operation.And blood lactate dehydrogenase(LDH),creatine kinase(CK),creatine phosphate kinase isoforms MB(CK-MB),cardiac troponin T(cTnT),serum B natriuretic peptide(pro-BNP)in plasma were detected by enzyme-linked immunosorbent assay(ELISA).Interleukin 10(IL-10),Interleukin 6(IL-6)and tumor necrosis factor α(TNF-α)were assayed.Pathological changes of myocardial tissue were detected by HE and Masson.The expression of microRNA-214 was detected by RT-PCR.The expression of Bax,Caspase-3,BCl2,PI3K,Akt,FoxO1 was detected by Western Blot.Results Compared with Sham group,IRI group showed a significantly increases in myocardial injury parameters of LDH,CK,IL-6 and TNF concentration in plasma,and a significantly reduced concentration of IL-10(P<0.05).And compared with Sham group,MI group showed a significantly increased expression of microRNA-214(P<0.05)and showed a significantly increased myocardial parameters of Bax,Caspase-3,PI3K,Akt protein,and a decreased level of BCl2,FoxO1(P<0.05).Compared with IRI group,microRNA-21 group showed a reduced myocardial ischemia-reperfusion-induced myocardial injury in rats and a reduced plasma concentration of LDH,CK,IL-6 and TNF-alpha,a inhibited expression of caspase-3,Bax,myocardial PI3K and Akt,and a promoted expression of BCl2 and FoxO1 protein(P<0.05).Conclusions MicroRNA-214 reduces the myocardial injury induced by myocardial ischemia-reperfusion through PI3K/Akt signaling pathway.

Killer effect of natural killer cells combined with tamoxifen on breast cancer cells in vitro and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the synergistic killer effect of natural killer cells(NK cells) combined with tamoxifen(TAM) on breast cancer cells(BCC)through the experiment in vitro,and to explore its mechanism.Methods:Three kinds of BCC with different receptor expression levels were selected for the experiment.Blank control group, different concentrations of TAM groups and different time groups were set up.MTT assay was used to detect the inhibitory rates of proliferation of cells,and the final experiment concentration of 5 μmol·L-1 was determined.The cells were divided into natural-release group,largest-release group,TAM group,NK cells group, and combined-experimental group(BCC+NK cells+TAM),and the synergistic killer effect of NK cells combined with TAM in different effector-target ratios were detected with Calcein-AM release assay.In ELISA assay the cells were divided into blank control group (NK cells),NK cells+TAM group, NK cells+BCC group and combined-experimental group,and the levels of IFN-γ and TNF-α in the NK cells in various groups were measured.In flow cytometry detection the cells were divided into blank control group (NK cells),NK cells+TAM group,NK cells+ BCC group, and combined-experimental group,and the expression levels of NKp46,CD158a,CD158b,CD158b2,and CD158e were determined;while the cells were divided into blank control group (BCC),BCC+TAM group,BCC + NK cells group, and combined-experimental group,and the expression levels of the MICA,ULBP1 and ULBP2 were detected.Results: The MTT assay results showed that the inhibitory rates of proliferation of 3 kinds of BCC had obvious time-and concentration-dependence (P<0.05).The Calcein-AM release assay results showed that the killing-rates of BCC in TAM groups were increased with the increase effector-target ratios of compared with NK cells group;and the killing-rate in combined experimental group was obviously higher than those in NK cells and TAM groups(P<0.05).The ELISA assay results showed that the levels of TNF-α and IFN-γ of NK cells in various experimental groups with BCC or not were increased compared with blank control group(P<0.05 or P<0.01);the levels were significantly increased when combined with TAM (P<0.05).The flow cytometry results showed that the NKp46 expression levels in various experimental groups were elevated compared with blank control group(P<0.05);the expression levels of CD158a, CD158b,CD158b2, and CD158e were significantly decreased(P<0.05);the expression levels MICA,ULBP1, and ULBP2 in BCC were significantly increased (P<0.05).Conclusion:The NK cells combined with TAM has the synergistic killer effect on the BCC in vitro.The synergetic mechanism may be as follows: TAM could increase the secretion of TNF-α and IFN-γ of NK cells to enhance their cytotoxicity;TAM also could up-regulate the expression levels of activating receptors and activating ligands,and down-regulate the expression levels of inhibitory receptors to increase the killing ability of NK cells.

A multicenter, randomized, controlled, phase Ⅲ clinical study of PEG-rhG-CSF for preventing chemotherapy-induced neutropenia in patients with breast cancer and non-small cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the safety and efficacy of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing chemotherapy-induced neutropenia in patients with breast cancer and non-small cell lung cancer (NSCLC), and to provide the basis for clinical application.</p><p><b>METHODS</b>According to the principle of open-label, randomized, parallel-group controlled clinical trial, all patients were randomized by 1∶1∶1 into three groups to receive PEG-rhG-CSF 100 μg/kg, PEG-rhG-CSF 6 mg, or rhG-CSF 5 μg/kg, respectively. The patients with breast cancer received two chemotherapy cycles, and the NSCLC patients received 1-2 cycles of chemotherapy according to their condition. All patients were treated with the combination chemotherapy of TAC (docetaxel+ epirubicin+ cyclophosphamide) or TA (docetaxel+ epirubicin), or the chemotherapy of docetaxel combined with carboplatin, with a 21 day cycle.</p><p><b>RESULTS</b>The duration of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg and PEG-rhG-CSF 6 mg groups were similar with that in the rhG-CSF 5 μg/kg group (P>0.05 for all). The incidence rate of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group, and G-CSF 5 μg/kg group were 69.7%, 68.4%, and 69.5%, respectively, with a non-significant difference among the three groups (P=0.963). The incidence rate of febrile neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg/kg group were 6.1%, 6.4%, and 5.5%, respectively, showing no significant difference among them (P=0.935). The incidence rate of adverse events in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg / kg group were 6.7%, 4.1%, and 5.5%, respectively, showing a non-significant difference among them (P=0.581).</p><p><b>CONCLUSIONS</b>In patients with breast cancer and non-small cell lung cancer (NSCLC) undergoing TAC/TA chemotherapy, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF at 48 hours after chemotherapy show definite therapeutic effect with a low incidence of adverse events and mild adverse reactions. Compared with the continuous daily injection of rhG-CSF 5 μg/kg/d, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF has similar effect and is more advantageous in preventing chemotherapy-induced neutropenia.</p>

Ad-hVEGF165 reverses homocysteine-induced endothelial dysfunction by regulating nitric oxide system / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of Ad-hVEGF165 on the endothelial cells dysfunction induced by homocysteine (Hcy) and related molecular mechanisms.</p><p><b>METHODS</b>Human umbilical vein endothelial cells CRL-1730 were treated with Hcy at different concentrations (0, 0.05, 1.00 mmol/L) for 24 h. The same concentration of Hcy, Ad-Track and Ad-hVEGF165 were added to the cells in the following groups: blank group, Hcy0.05 group, Hcy1.00 group, Ad-Track group, Hcy0.05+Ad-Track group, Hcy1.00+Ad-Track group, Ad-hVEGF165 group, Hcy0.05+Ad-hVEGF165 group, Hcy1.00+ Ad-hVEGF165 group for 48 h. The mRNA and protein expressions of eNOS and DDAH2 were detected by real-time PCR and Western blot. The correlations of mRNA and protein expressions between endothelial nitric oxide synthase (eNOS) and dimethylarginine dimthylaminohydrolase (DDAH)2 were evaluated by Pearson correlation analysis.</p><p><b>RESULTS</b>Compared with blank group and Ad-hVEGF165 group, the mRNA and protein expressions of eNOS were decreased in Hcy0.05 group and Hcy0.05+Ad-hVEGF165 group (both P < 0.05), and the mRNA and protein expressions of DDAH2 in cells treated with 0.05 mmol/L and 1.00 mmol/L Hcy were reduced as well (all P < 0.05). DDAH2 mRNA and protein expression are increased (all P < 0.05) in Ad-hVEGF165 group compared with the blank group and Ad-Track, Hcy0.05 + Ad-hVEGF165 and Hcy0.05 group compared with Hcy0.05+Ad-Track group, Hcy1.00+Ad-hVEGF165 and Hcy1.00 group compared with Hcy1.00+Ad-Track group. The mRNA and protein expressions of eNOS and DDAH2 were uncorrelated under the effect of Hcy (r = 0.057 and 0.449, both P > 0.05) and VEGF (r = 0.284 and 0.432, both P > 0.05).</p><p><b>CONCLUSION</b>Recombinant adenovirus Ad-hVEGF165 could reverse Hcy-induced endothelial cells dysfunction via upregulating the expressions of eNOS and DDAH2.</p>

Lymph node metastasis and prognostic analysis of 354 cases of T1 breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the characteristics of lymph node metastasis and prognosis in patients with T1 breast cancer.</p><p><b>METHODS</b>The clinicopathological data of 354 patients with T1 breast cancer after standard treatment from March 2007 to September 2011 were collected to analyze the relationship between the clinical characteristics of T1 breast cancer, lymph node metastasis and prognostic features.</p><p><b>RESULTS</b>In the 354 patients with T1 breast cancer, 105 patients (29.7%) had lymph node metastasis, among them 73 cases (69.5%) had 1-3 lymph node metastasis, and 32 cases (30.5%) had more than 4 lymph node metastasis. The lymph node metastasis rate was 8.3% in T1a patients, 39.7% in T1b patients, and 30.4% in T1c cases (P = 0.005). Pairwise comparison showed that the difference of lymph node metastasis rate between T1a, T1b and T1c patients was statistically significant (P = 0.001 and P = 0.006, respectively). The difference of lymph node metastasis rates in T1b and T1c patients was statistically insignificant (P = 0.171). In the 354 patients of T1 breast cancer, 92 patients had vascular tumor thrombi and their lymph node metastasis rate was 71.7%, while the lymph node metastasis rate in 262 patients without vascular tumor thrombus was 14.9% (P < 0.001). The median follow-up was 49 months (range 27-81 months). 12 patients developed recurrence, and 3 patients died, one of them died of cerebrovascular accident. The 4-year disease-free survival for all patients was 96.6%, and the 4-year overall survival rate was 99.2%.</p><p><b>CONCLUSIONS</b>There is a correlation between vascular tumor thrombus, tumor size and lymph node metastasis rate. The lymph node metastasis rate is lower in T1a patients and relatively higher in T1b/c patients. Compared with patients without vascular tumor thrombus, the T1 breast cancer patients with vascular tumor thrombi have a higher lymph node metastasis rate. Generally speaking, there is a still good prognosis in patients with T1 breast cancer.</p>

Effect of 20 (S)-ginsenoside Rg3 on the proliferation inhibition and secretion of vascular endothelial growth factor of multiple myeloma cell line U266 / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of 20 (S)-ginsenoside Rg3 [20 (S)-Rg3] on the proliferation inhibition and secretion of vascular endothelial growth factor (VEGF) of multiple myeloma (MM) cell line U266.</p><p><b>METHODS</b>The proliferation inhibition rate of U266 cells after treatment with different doses of 20 (S)-Rg3 was detected by MTT method, the cell cycle and apoptosis by flow cytometry, the expression of apoptosis related proteins of caspase-3, 8 and 9 by Western blot, VEGF concentration in the culture supernatant by ELISA.</p><p><b>RESULTS</b>It showed that 20 (S)-Rg3 could inhibit the proliferation of U266 in a dose-dependent manner (P<0.05) with IC50 of (71.07 ± 2.63)μmol/L and (44.06 ± 3.98) μmol/L at 24 h and 48 h, respectively. VEGF concentration in the culture supernatant showed a dosedependent reduction (P<0.05), decreased from (419.93 ± 36.76) pg/106 cells in the control group to (314.82 ± 27.05) pg/106 cells in 80 μmol/L 20 (S)-Rg3 treated group by ELISA assay. Flow cytometry with Annexin-V/PI double staining revealed that 20(S)-Rg3 may induce U266 cells apoptosis in a concentration-dependent manner from (0.51 ± 0.05)% at control group to (8.32 ± 0.83)%, (10.72 ± 1.29)% and (15.27 ± 2.26)% at 20, 40 and 80 μmol/L treatment groups, respectively (P<0.05). Flow cytometry with PI staining showed that the ratio of cells in G0/G1 phase increased from (49.11 ± 1.71)% to (52.72 ± 7.75)%, (60.29 ± 5.76)% and (61.81 ± 3.46)%, respectively (P<0.05). Western blot analysis indicated that the expression of caspase-3, 8 and 9 declined, and that of cleaved-caspase-3, 8 and 9 significantly increased (P<0.05) with 20 (S)-Rg3 concentration increased.</p><p><b>CONCLUSION</b>20(S)-Rg3 can inhibit the proliferation of U266 cells by cell cycle arrest in G1 phase and induce cell apoptosis by increasing the expressions of cleaved-caspase-3, -8 and -9. It can also inhibit VEGF secretion of U266 cells, which makes it a potential agent for multiple myeloma therapy.</p>

Efficacy of 27 patients with multiple myeloma / 白血病·淋巴瘤

Objective To evaluate the efficacy and safety of bortezomib-based chemotherapy and MPT regimen in the MM patients who were newly diagnosed or relapsed/refractory. Methods Twenty-seven MM patients were treated with bortezomib-based chemotherapy, median cycles:3 (range 1-5 cycles). Other 30patients received MPT chemotherapy. EBMT and WHO criteria were used to evaluate the therapeutic effects and the adverse effects, respectively. Results Bortezomib group: 21 patients (77.8 %) showed effects after the first cycle chemotherapy and 24 patients (88.8 %) showed effects after the whole therapy. In wich, 15 patients(94.0 %) and 9 patients (82.0 %) were newly diagnosed and relapsed/refractory, respectively. MPT group: 15patients (50.0 %) showed effects after the whole therapy. In wich, 12 patients (44.0 %) were newly diagnosed.And the other 3 were relapsed/refractory patients. The ORR in Bortezomib group was better than MPT group (P ＜0.05). The incidence of peripheral neuropathy, herpes and Ⅲ - Ⅳ grade thrombocytopenia in the bortezomib group was 10 patients (37.0 %), 7patients (26.0 %), 10 patients (37.0 %) respectively,and they were more common than MPT group, but the incidence of Ⅲ-Ⅳgrade anemia was 21 patients (70.0 %) and more comumom in the MPT group. The theraputic efficacy of bortezomib for renal insufficiency and normal renal function patients was similar, and no significant increase in all kinds of adverse effects. In MPT group,there were 4 patients with renal insufficiency, the serum level of creatinine in the 3 patients returned to normal after 5 cycles therapy. Conclusion Bortezomib-based chemotherapy is more effective than MPT regimen in the treatment of MM. The newly diagnosed, relapsed/ refractory and with renal insufficiency patients all can benefit from it. The adverse effects are mild and with better tolerance.

Effect of bone marrow stromal cells transfeted by human sFlt-1 gene on growth of K562 cells / 吉林大学学报(医学版)

Objective To get the protein expression of sFlt-1 by transfection and to investigate the effects of sFlt-1 gene transfection on the growth of K562 cells.Methods The recombinant plasmid pcDNA3-sFlt-1D4 was constructed.The recombinant plasmid pcDNA3-sFlt-1D4 was transfected into the bone marrow stromal cells by Lipofectamine 2000,which was identified by RT-PCR,ELISA and MTT.Results The transfection efficiency identified by flow cytometry was 9.27%.The protein expression of sFlt-1D4 was found in the culture supernatant 24 h,48 h and 4 weeks after transfetion by ELISA and the expression concentrations were(0.104?0.078),(0.158?0.022) and(0.171?0.069) ?g?L-1,respectively.The content of VEGF secreted by K562 culturing with transfectant cells culture supernatant was reduced compared with control.The inhibitoy rates on the proliferation of K562 cells via MTT assay were 9.41%?4.71%,23.63%?7.50%,and 33.13%?6.93%,respectively.Conclusion The bone marrow stromal cells transfected with recombinant plasmid pcDNA3-sFlt-1D4 could secrete sFlt-1D4 and inhibit the proliferation of K562 cells.