BACKGROUND:Endothelial progenitor cells are recruited into local vascular injury under the injury-induced stimulation, and then differentiate into mature endothelial cells that are thereby involved in angiogenesis and endothelial repair. OBJECTIVE:To investigate whether endothelial progenitor cells can al eviate renal injury and improve renal function of ischemia/reperfusion injury (I/R) rats. METHODS:Peripheral blood samples extracted from Sprague-Dawley rats were used to isolate and culture endothelial progenitor cells using density gradient centrifugation. Twenty-two male Sprague-Dawley rats were randomized to three groups:I/R group, normal control group, and endothelial progenitor cells group. In the I/R and endothelial progenitor cells groups, the right kidney was removed and the renal artery and vein of the left kidney were occluded for 40 minutes to establish I/R models in the rats, and then endothelial progenitor cells (5×109/L, total y 1 mL) or solvent was transplanted via the artery of the left kidney into the left kidney. In the normal control group, the experimental procedure was same as that in the I/R group except for occlusion of the artery and vein of the left kidney. Renal and blood samples from three groups were col ected at day 1 after operation. Peripheral blood CD34 and vascular endoethelial growth factor receptor 2 expressions were determined using flow cytometry and immunofluorescence methods, serum creatinine and urea nitrogen were tested, and immunohistochemistry observation was used for CD34 observation. RESULTS AND CONCLUSION:Compared with the normal control group, serum creatinine and urea nitrogen levels were significantly increased, and tubulointerstitial CD34 expression was decreased in the I/R group (P<0.05). Endothelial progenitor cells treatment largely decreased the levels of serum creatinine and urea nitrogen, and increased CD34 expression (P<0.05). These findings indicate that transplantation of endothelial progenitor cells contributes to renal protection in I/R rats.

Objective To investigate the clinical values of Gemstone Spectral Imaging coronary angiography with analysis of the relationship between plaque character and angina pectoris types.Methods Two hundred cases of hospitalized patients were collected from January 2015 to December 2016 in the Department of Cardiovascular Disease,Chenzhou City First People's Hospital,who were diagnosed as coronary heart disease.According to the clinical data,stable angina pectoris (SA) and unstable angina pectoris (UA),coronary CT scan was performed for each patient to evaluate the relationship between the nature of atherosclerotic plaques and the classification of angina pectoris.SPSS 19.0 software was used for statistical analysis.Results By analyzing the correlation between SA and UA,SA calcification plaque was accounted for 63.2％,soft plaque 17.4％,mixed plaque 19.4％;UA calcification plaque was accounted for 37.5％;Soft plaque was accounted for 42.3％;and mixed plate was accounted for 20.2％.The difference was statistically significant (P ＜ 0.05).In the SA group,all of the stenotic lesions were mainly calcified plaques.There was no significant difference in the degree of severe stenosis between the different plaques in the SA group (P ＞ 0.05).In UA group,soft plaques were the main lesion of mild and moderate stenosis of coronary artery,and calcified plaques were the main stenosis in severe stenosis.There were significant differences in the degree of coronary artery stenosis between different plaques in UA group (P ＜ O.05).Conclusions The 128-slice spiral CT can accurately evaluated the relationship between the nature of coronary plaque and the type of angina pectoris,which is of great value in the early warning and angina pectoris classification of high-risk patients with coronary heart disease.

The contribution of particles to cardiovascular mortality and morbidity has been enlightened by epidemiologic and experimental studies. However, adverse biological effects of the particles with different sizes on cardiovascular cells have not been well recognized. In this study, sub-cultured human umbilical vein endothelial cells (HUVECs) were exposed to increasing concentrations of pure quartz particles (DQ) of three sizes (DQPM1, <1 μm; DQPM3-5, 3-5 μm; DQPM5, 5 μm) and carbon black particles of two sizes (CB0.1, <0.1 μm; CB1, <1 μm) for 24 h. Cytotoxicity was estimated by measuring the activity of lactate dehydrogenase (LDH) and cell viability. Nitric oxide (NO) generation and cytokines (TNF-α and IL-1β) releases were analyzed by using NO assay and enzyme-linked immunoabsorbent assay (ELISA), respectively. It was found that both particles induced adverse biological effects on HUVECs in a dose-dependent manner. The size of particle directly influenced the biological activity. For quartz, the smaller particles induced stronger cytotoxicity and higher levels of cytokine responses than those particles of big size. For carbon black particles, CB0.1 was more capable of inducing adverse responses on HUVECs than CB1 only at lower particle concentrations, in contrast to those at higher concentrations. Meanwhile, our data also revealed that quartz particles performed stronger cell damage and produced higher levels of TNF-α than carbon black particles, even if particles size was similar. In conclusion, particle size as well as particle composition should be both considered in assessing vascular endothelial cells injury and inflammation responses induced by particles.

OBJECTIVETo investigate the anti-tumor activity of CCL21-exCD40L eukaryotic expression vector.

METHODSCCL21-exCD40L fusion gene were constructed by overlap PCR connecting CCL21 and exCD40L through a flexible linker (Gly3Ser)4, and then was cloned into expression vector pcDNA3.1(+). pcDNA3.1(+)/CCL21 and pcDNA3.1(+)/exCD were constructed as negative control. Wsestern blot was used to identify the fusion protein. CHO cells was transfected with pcDNA3.1(+)/CCL21-exCD, pcDNA3.1(+)/CCL21 and pcDNA3.1(+), respectively. The chemotatic function of the expressed product was detected by Transwell method and its anti-tumor activity was tested with vivo transfection.

RESULTSGene sequencing and restrictive digestion proved the successful construction of pcDNA3.1(+)/CCL21-exCD40L,and its expression was conformed by western blot. The transfectant supernantes of pcDNA3.1(+)/CCL21-exCD40 group had a significant chmotactic function to DCs, of which the cell numbers passing through the film was 14.95 times of blank control every high power microscope visual field. After tumor orthotoic injection of plasmid carrying fusion gene in Balb/c mouse, the tumor mass reduced remarkablely, and all the mouse in fusion gene group survived after 4 weeks.

CONCLUSIONCCL21-exCD40L fusion protein had a remarkable function to DCs and it can inhibit tumor growth and prolong the mouse survival time, which is more effective than all control group.

Animals ; CD40 Ligand ; genetics ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Chemokine CCL21 ; genetics ; pharmacology ; Colonic Neoplasms ; therapy ; Cricetulus ; Dendritic Cells ; drug effects ; physiology ; Genetic Therapy ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; pharmacology

In order to identify the chemical constituents of Yushu tablets comprehensively, we studied the chemical constituents of CHCl3 extract from Yushu tablets by the ultra performance liquid chromatography-electrospray ionization-ion trap-time of flight mass spectrometry (UPLC-ESI-IT-TOF/MS). It showed that there were more than 100 compounds separated, and forty-nine peaks among these were identified on the basis of high resolution mass spectrometry data and literature data reported. Determination of twelve peaks was further confirmed by standard substances. These components assigned to the different plant sources mainly included phenylpropanoids, triterpenoids, quinones and m-trihydroxybenzene compounds. By analyzing the chemical components of CHCl3 extract from compound Chinese medicine Yushu tablets comprehensively, this study provided the foundation for studying chemical components, pharmacodynamic substance and quality control of Yushu tablets.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drugs, Chinese Herbal ; analysis ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Tandem Mass Spectrometry

Objective: To analyze the association of thrombocytosis with the prognosis of patients with gastric cancer.Methods: The clinical materials of 782 patients with gastric cancer who underwent initial surgery in our hospital between January 1995 and December 1999 were retrospectively analyzed.Kaplan- Meier and Log-Rank test were used to analyze the data.Prognostic factors were analyzed by multivariate Cox proportional hazards model.Results: Thrombocytosis oc-curred in 11.4% (87/782) patients.The platelet level was not significantly different among patients of different gender, tumor stage, and histological differentiation (P>0.05).However, a significant difference was observed in the platelet level among patients with different age and surgical approach (P<0.05).The 1-, 3-, and 5-year survival rates were 75.0%, 40.1% and 28.9% in patients without thrombocytosis and 52.8%, 16.9%, and 13.5% in patients with thrombocytosis (P=0.002).Univari-ate analysis showed that histological differentiation, pathological stage, surgical approach and thrombocytosis significantly affected the survival of patients.While age and gender had no significant impact on patient survival.Multivariate analysis showed that pathological stage, surgical approach, and thrombocytosis were independent prognostic factors for gastric can-cer.The relative risk of death of patients with thrombocytosis was elevated by 1.454 times (RR=1.454, 95% CI: 1.135～1.861, P=0.005).Conclusion: Thrombocytosis is an independent prognostic indicator for the survival of initially treated pa-tients with gastric cancer.

Objective To explore the correlation between plasma Klotho and FGF23 levels with senile hypertension .Methods Totally 180 cases of hypertension aged more than 60 years old were selected as the research group and divided into the grade 1 hypertension group(60 cases) ,grade 2 hypertension group(60 cases) and grade 3 hypertension group(60 cases) .The age and pro‐portion of male to female were basically consistent .60 elderly people without hypertension were selected as the control group .The level of plasma Klotho and FGF23 protein were tested by ELISA .The lipid level was detected by the automatic biochemical analy‐zer .The detection results were compared with those in the control group for analyzing their correlation .Results Plasma Klotho protein level in each hypertension group was lower than that in the control group ,while plasma FGF23 protein level and BMI were significantly higher than those in the control group(P<0 .05) .With the blood pressure level increase ,plasma Klotho protein level was gradually decreased ,while FGF23 protein level was gradually increased .The negative correlation was found between plasma Klotho protein and FGF23 protein (r= -0 .282 ,P=0 .001) .Plasma Klotho protein and FGF23 protein had no obvious correlation with BMI (r= -0 .063 ,-0 .098 ,P>0 .05) .Conclusion Plasma Klotho protein and FGF23 protein levels have obvious correlation with the blood pressure level ,which may jointly participate in the occurrence and development of hypertension .

Objective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.

Objective To investigate the role of heme oxygenase-1 ( HO-1 ) and carbon monoxide ( CO) in children with mycoplasma pneumoniae infection. Methods A total of 198 cases with mycoplasma pneumoniae infection under 3 years were enrolled in this study. According to whether with wheezing,all cases were divided into wheezing group( n=58 ) and no wheezing group ( n=140 ) . According to having hypox-emia,wheezing group was divided into mild wheezing group(n=34) and severe wheezing group(n=24). And no wheezing group was set as control group. The serum COHb levels were measured by dual-wavelength spectrophotometry. The serum HO-1 were measured by ELISA. Results The levels of serum HO-1 and COHb% in severe wheezing group were ( 2 734. 10 ± 707. 59 ) ng/L, ( 3. 88 ± 0. 83 )%, and ( 1 130. 03 ± 671. 02) ng/L,(1. 16 ± 0. 63)% in mild wheezing group,and(931. 32 ± 451. 67) ng/L,(1. 04 ± 0. 37)% in no wheezing group,respectively. There were significant differences in serum HO-1 and COHb% between se-vere wheezing group and mild wheezing group(P<0. 01,respectively),and there were no significan differ-ences in serum HO-1 and COHb% between mild wheezing group and no wheezing group( P>0. 05,respec-tively). There was significant positive correlation between serum HO-1 and COHb%(r=0. 875,P<0. 01). Conclusion With mycoplasma pneumoniae infection,the expression of HO-1 and COHb% increase by the wheezing disease progression. HO-1 and CO may participate in the development process of infantile wheezing diseases.