Neuroprotective effect of surround needling combined with acupoint injection on acute herpetic neuralgia / 中国针灸

OBJECTIVE@#To explore the effect and mechanism of surround needling combined with acupoint injection on acute herpetic neuralgia (AHN).@*METHODS@#Ninety-nine patients with T-T segment AHN were randomly divided into 3 groups, 33 cases in each group, including 2 cases dropped off in the surround needling group, 4 cases dropped off in the acupoint injection group, and 3 cases dropped off in the combined group. Oral valacyclovir was given in each group, 0.3 g each time, 2 times a day for 10 days. Oblique insertion of needle used at points around the herpes in the surround needling group, and continuous wave was stimulated to tolerance for 20 min; the same acupoints were selected as the surround needling group, stimulated with the mixture injection of mecobalamin and lidocaine in the acupoint injection group; After the surround needling, acupoint injection was performed in the combined group. The treatment was given once a day, 14 times for a course, and one course was needed in all groups. The skin healing conditions (blistering, crusting, and dislocation time) of each group were compared after treatment. The pain scores, pain area and quality of life scores in each group were observed before and after treatment. The levels of neuron specific enolase (NSE), substance P (SP) and calcitonin gene-related peptide (CGRP) in the local blister fluid were measured before and after treatment in all groups.@*RESULTS@#The blistering, crusting and dislocation time in the combined group were earlier than the other two groups (all <0.05). The pain score and pain area in the each group were significantly lower than those before treatment, and the quality of life score was significantly higher than that before treatment (all <0.05). The improvements of pain score and quality of life score in the combined group were more obvious than the other two groups (all <0.05). After treatment, the levels of NSE, SP and CGRP in the local blister fluid in each groups were significantly lower than those before treatment (all <0.05). The indexes in the combined group were significantly lower than those in the other two groups (all <0.05).@*CONCLUSION@#Both surround needling and acupoint injection have an adjuvant effect on AHN. The combination of the two is better, the skin is healed quickly, the analgesia is significant, and the contents of local NSE, SP and CGRP are significantly decreased. The mechanism of action is to exert neuroprotective effects.

The value of joint detection of sTREM-1 and PCT in the early diagnosis of children with sepsis / 中国医学装备

Objective: To investigate the value of joint detection of soluble triggering receptor expresses on myeloid cells-1(sTREM-1) and procalcitonin (PCT) in the early diagnosis of children with sepsis. Methods: 78 children with sepsis were selected into the sepsis group, 23 children with common infection were selected into the normal infection group. In addition, 25 healthy children selected into the health control group. The levels of sTREM-1, PCT, and C reactive protein (CRP) among the three groups were compared, respectively. And then, the sepsis group were further divided into general sepsis subgroup (32 cases), severe sepsis subgroup (26 cases) and septic shock subgroup (20 cases) according to the degree of sepsis. The levels of sTREM-1, PCT and CRP among the three sepsis subgroups were compared. And the receiver operating characteristic (ROC) curve was adopted to analyze the value that diagnosed children with sepsis by using the three indicators. Results: The levels of sTREM-1, PCT and CRP of sepsis group were significantly higher than those of common infection group and health control group (t=22.071, t=21.508, t=17.870, t=55.167, t=52.070, t=30.359, P<0.05). The differences of sTREM-1 and PCT among various sepsis subgroups were significant (H=22.082, H=39.449, P<0.05), but the difference of CRP level between septic shock subgroup and severe sepsis subgroup was no significant. As the compared result of AUC of ROC of diagnosing sepsis, the AUC of sTREM-1 was maximum (0.88), and its 95% confidence interval (CI) was 0.78-0.98. At the optimum cutoff value of sTREM-1, the sensitivity and specificity were 83.33% and 68%, respectively, and they were higher than those of PCT and CRP, respectively. Besides, the cutoff values of sTREM-1 and PCT were used as standard to carry out joint diagnosis for children with sepsis, and the sensitivity and specificity were 91.03% and 64%, respectively, at this joint diagnosis. Conclusion: The joint detection of sTREM-1 and PCT has higher sensitivity in the early diagnosis of children with sepsis and it has a certain clinical application value.

The study of the protective effect of panoxadiol on Alzheimer’s disease cells based on network pharmacology / 中国药理学通报

Aim To explore the therapeutic effects of main active compounds of panaxadiol ( PD ) in on Alzheimer’s disease ( AD) via network pharmacologi-cal analysis and Mmolecular docking. Methods A to-tal of 107 prescriptions for AD treatment were screened by using network pharmacology, screening for the high-est frequency of ginseng and its target for AD. Use mo-lecular docking technology was used to find components with the highest score for non-receptor tyrosine kinase ( FYN) docking. Then we successfully estimatedestab-lished AD cell model with overexpressinged APP pro-teins in vitro. Next,the cell viability was detected by MTT assay,the cell damage was detected by LDH as-say,the apoptosis and intracellular Ca2+concentration were detected by flow cytometry, and phosphorylated FYN protein expression was detected by Western blot detection of . phosphorylated FYN protein expression. Results Eighteen active components of Gginseng and 29 AD-related targets were screened by the method of network pharmacology. The results of molecular doc-king showed that PD had strong binding effects with FYN. The results showed that PD could increase the survival rate of cells,reduce the release of LDH,reduce apoptosis,and improve AD cells’ intracellular Ca2+o-verload and reduce the expression of FYN-Y416 pro-tein. Conclusion The experimental results of network pharmacology were are verified and the protective effect of PD on AD may be related to inhibition of FYN signa-ling pathway.

Research progress in treatment of cryptogenic stroke associated with patent foramen ovale / 解放军医学杂志

Patent foramen ovale (PFO) is a common congenital heart disease that can cause cryptogenic stroke through paradoxical embolization.For patients with PFO combined with cryptogenic stroke,whether anticoagulant therapy is superior to antiplatelet therapy in the prevention of recurrent stroke? And whether PFO closure can significantly reduce the risk of stroke recurrence compared with medical therapy alone? All those raised the clinical problems to be solved urgently.The advances in treatment of cryptogenic stroke associated with PFO are herewith summarized in present paper by reviewing randomized trials,meta-analyses and the guidelines or expert consensus about PFO and cryptogenic stroke.

Role of human liver and intestine esterases in drug design and development / 药学学报

Human carboxylesterase (CES) and arylacetamide deacetylase (AADAC) are important numbers of the serine esterase superfamily. They are involved in hydrolytic procedure of human endogenous cholesteryl esters, as well as drug metabolism, activation and detoxication. They are closely related to the personalized medication of drugs, especially for prodrugs. This review summarizes their structure and distribution, metabolic characteristics and research progress in recent years, which will provide a reference for new drug development and rational drug design.

Improving Asymptomatic Left Ventricular Diastolic Dysfunction in Postmenopausal Hypertensive Women with Metabolic Syndrome: A Prospective, Open-Labeled, Randomized Controlled Trial / 中华医学杂志(英文版)

<p><b>Background</b>Postmenopausal women with metabolic syndrome (MetS) have increased cardiovascular morbidity and left ventricular diastolic dysfunction (LVDD). The various protective effects of astragalus membranaceus (AM) have been described in previous studies. Therefore, this study aimed to evaluate the effects of different doses of AM on diastolic function in postmenopausal hypertensive women with MetS.</p><p><b>Methods</b>This was a prospective, randomized controlled study. The postmenopausal hypertensive patients with MetS were enrolled from Lanzhou University Second Hospital from March 2014 to April 2015. Patients were divided into three groups: control group (received conventional medical treatment), AM Group 1 (received AM capsules at 5 g/d additionally), and AM Group 2 (received AM capsules at 10 g/d additionally). Echocardiographic and clinical characteristics were evaluated before and 12 months after treatment. Quantitative data were analyzed using unpaired t-test, analysis of variance, and multiple linear regression analysis.</p><p><b>Results</b>A total of 154 patients were subjected to final analysis. In the AM Group 2, significant improvements were noted in diastolic function 12 months after treatment than those of the control group, including the early diastolic mitral annular velocity (E'; 0.065 ± 0.007 m/s vs. 0.061 ± 0.008 m/s, P = 0.014), the ratio of the early diastolic mitral peak flow velocity to the late diastolic mitral peak flow velocity (E/A; 0.81 ± 0.05 vs. 0.80 ± 0.06, P = 0.012), the ratio of E' to the late diastolic mitral annular velocity (E'/A'; 0.56 ± 0.12 vs. 0.51 ± 0.13, P = 0.048), and the ratio of the early diastolic mitral peak flow velocity (E) to E' (E/E'; 10.70 ± 1.30 vs. 11.37 ± 1.73, P = 0.031). After treatment, E/E' (10.70 ± 1.30 vs. 11.24 ± 1.56, P = 0.021), deceleration time (DT; 261.49 ± 44.41 ms vs. 268.74 ± 53.87 ms, P = 0.046), and E'/A' (0.56 ± 0.12 vs. 0.52 ± 0.13, P = 0.019) values improved more significantly than those of AM Group 2 before treatment. Besides, waist circumference was positively correlated with E' (r = 0.472; P = 0.003) and E'/A' (r = 0.321; P = 0.047). In addition, the waist-to-hip ratio was a significant predictor of DT (r = 0.276; P = 0.041), E' (r = -0.590; P < 0.001), E/E' (r = 0.454; P = 0.004), and E'/A' (r = -0.377; P = 0.018).</p><p><b>Conclusions</b>Conventional medical plus AM therapy improved diastolic function. Moreover, WC and WHR might be risk factors for LVDD.</p><p><b>Chinese Clinical Trial Register</b>ChiCTR-TRC-11001747. http://www.chictr.org.cn/showprojen.aspx?proj=7798.</p>

Improving Asymptomatic Left Ventricular Diastolic Dysfunction in Postmenopausal Hypertensive Women with Metabolic Syndrome: A Prospective, Open-Labeled, Randomized Controlled Trial / 中华医学杂志(英文版)

<p><b>Background</b>Postmenopausal women with metabolic syndrome (MetS) have increased cardiovascular morbidity and left ventricular diastolic dysfunction (LVDD). The various protective effects of astragalus membranaceus (AM) have been described in previous studies. Therefore, this study aimed to evaluate the effects of different doses of AM on diastolic function in postmenopausal hypertensive women with MetS.</p><p><b>Methods</b>This was a prospective, randomized controlled study. The postmenopausal hypertensive patients with MetS were enrolled from Lanzhou University Second Hospital from March 2014 to April 2015. Patients were divided into three groups: control group (received conventional medical treatment), AM Group 1 (received AM capsules at 5 g/d additionally), and AM Group 2 (received AM capsules at 10 g/d additionally). Echocardiographic and clinical characteristics were evaluated before and 12 months after treatment. Quantitative data were analyzed using unpaired t-test, analysis of variance, and multiple linear regression analysis.</p><p><b>Results</b>A total of 154 patients were subjected to final analysis. In the AM Group 2, significant improvements were noted in diastolic function 12 months after treatment than those of the control group, including the early diastolic mitral annular velocity (E'; 0.065 ± 0.007 m/s vs. 0.061 ± 0.008 m/s, P = 0.014), the ratio of the early diastolic mitral peak flow velocity to the late diastolic mitral peak flow velocity (E/A; 0.81 ± 0.05 vs. 0.80 ± 0.06, P = 0.012), the ratio of E' to the late diastolic mitral annular velocity (E'/A'; 0.56 ± 0.12 vs. 0.51 ± 0.13, P = 0.048), and the ratio of the early diastolic mitral peak flow velocity (E) to E' (E/E'; 10.70 ± 1.30 vs. 11.37 ± 1.73, P = 0.031). After treatment, E/E' (10.70 ± 1.30 vs. 11.24 ± 1.56, P = 0.021), deceleration time (DT; 261.49 ± 44.41 ms vs. 268.74 ± 53.87 ms, P = 0.046), and E'/A' (0.56 ± 0.12 vs. 0.52 ± 0.13, P = 0.019) values improved more significantly than those of AM Group 2 before treatment. Besides, waist circumference was positively correlated with E' (r = 0.472; P = 0.003) and E'/A' (r = 0.321; P = 0.047). In addition, the waist-to-hip ratio was a significant predictor of DT (r = 0.276; P = 0.041), E' (r = -0.590; P < 0.001), E/E' (r = 0.454; P = 0.004), and E'/A' (r = -0.377; P = 0.018).</p><p><b>Conclusions</b>Conventional medical plus AM therapy improved diastolic function. Moreover, WC and WHR might be risk factors for LVDD.</p><p><b>Chinese Clinical Trial Register</b>ChiCTR-TRC-11001747. http://www.chictr.org.cn/showprojen.aspx?proj=7798.</p>

Investigating the effects of compound WS090152 on non-alcoholic fatty liver in mice / 药学学报

To investigate the effects and the mechanism of compound WS090152 on non-alcoholic fatty liver (NAFL), the compound was administrated in C57BL/6J mice fed a high fat diet at 50 mg·kg-1 by lavage. The lipid accumulation in liver was determined by the content of hepatic triglyceride (TG) and the histological pathological analysis. The levels of body weight gain, serum total cholesterol (TC) and TG were measured to evaluate lipid metabolism. Insulin sensitivity was determined by glucose infusion rate (GIR) value in hyperinsulinemic-euglycemic clamp test. The expression of related proteins in liver was measured by Western blot. The effect on the target protein tyrosine phosphatase 1B (PTP1B) was assessed by the activity of recombinate human PTP1B in vitro, and by the expressions of PTP1B in vivo, respectively. The content of hepatic TG (PPPPP50 value of 0.34 μmol·L-1; the expression of PTP1B was significantly downregulated, and the phosphorylation of its downstream insulin receptor (IR) and AKT was upregulated by WS090152 administration in the livers of NAFL mice. The expression of hepatic lipogenesis-related proteins-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC) was attenuated. These results suggest that compound WS090152 can ameliorate NAFL by increasing insulin sensitivity and decreasing hepatic lipogenesis probably through inhibition of PTP1B.

Bioanalysis in the development of antibody-drug conjugates / 药学学报

Antibody-drug conjugates (ADCs) are complex molecules with cytotoxic small molecular drugs covalently bound to monoclonal antibodies via a linker and can improve the targeted drug delivery with minimizing the systemic toxicity. ADCs are heterogeneous mixtures with different drug-to-antibody ratios (DARs) and the DAR distribution is dynamically changing in vivo, therefore the bioanalysis of the ADCs is challenging. Enzyme-linked immunosorbent assay (ELISA) and LC-MS have been widely used in the ADCs bioanalytical assays. Just like other biotherapeutics, ADCs may elicit the host immune response and produce the anti-therapeutic antibody (ATA), which could affect its efficacy, pharmacokinetics, and safety. It is thereby important to investigate its immunogenicity in the ADC development. In this review, we summarized the ELISA- and LC-MS-based bioanalysis strategies for the development of ADCs, including DAR distribution, the determination of total antibody, conjugated antibody, conjugated drug, free drug, and ATA, with the expectation of providing insights and reference for the ADC development in China.

Effects of Mongolia Astragali Radix in protecting early cardiac and nephritic functions of patients of hypertension with metabolic syndrome / 中国中药杂志

To explore the effect of Mongolia Astragali Radix produced in Longxi of Gansu province in protecting cardiac and nephritic functions of patients of essential hypertension(EH) with metabolic syndrome(MetS). A total of two hundred and twenty-six EH patients with MetS aged above 18 were selected. Patients were randomly divided to control group(adopted conventional medical treatment), Astragali Radix group 1(added Astragali Radix capsules 10 g•d⁻¹ besides conventional medical treatment) and Astragali Radix group 2(added Astragali Radix capsules 5 g•d⁻¹ besides conventional medical treatment). Cardiac anatomy structure, cardiac systolic function and diastolic function were measured by M-mode echocardiography, two-dimensional echocardiography, Doppler echocardiographic determination and tissue Doppler imaging. The level of microalbuminuria(MAU) was evaluated by radioimmunoassay. In addition, the estimated glomerular filtration rate(eGFR) was calculated by modification of diet in renal disease (MDRD) formulas. The changes of relevant indicators for cardiac and nephritic functions before and after treatment were compared during the 12-month follow-up. The study protocol was registered at the website of Chinese clinical trial register and approved by the ethics committee of second hospital of Lanzhou university. Each patient was required to sign an informed consent. SPSS software was used for statistical analysis. According to the result, compare with before treatment, the three groups show no difference in efficacy of metablic indicators. Left ventricular end-systolic volume (ESV) and left ventricular end-systolic dimension (LVESd) of all patients were improved after treatment. However, there was no significant difference among the three groups. After the addition of Astragali Radix, the mitral flow velocity(Vp) of patients was improved to some extent(P<0.05). However, there was no significant difference among the three groups. Astragali Radix had a significant effect in reducing the MAU(P<0.05). Moreover, the MAU level of patients in Astragali Radix group 1 decreased more significantly than the other groups(P<0.05). Compared with conventional therapy, Astragali Radix combined with conventional therapy could improve cardiac structure, left ventricular systolic function, left ventricular diastolic function, and reduce the MAU to a certain extent in EH patients with MetS. Moreover, the effects of high-dose Astragali Radix are better than that of the low-dose Astragali Radix. However, the effect of Astragali Radix on EH patients with MetS shall be further observed to confirm its efficacy.

Prescription of Jingdan Yimin for treatment of metabolic syndrome / 中国中药杂志

Based on the theory of traditional Chinese medicine, modern methods for drug investigation such as molecular targets in vitro and effects in vivo were used to study the prescription of Jingdan Yimin(JD), including selection of raw materials, composition, proportion, and effective dose of the compounds for treatment of metabolic syndrome. The IRF mice models, characterized by insulin resistance and hypercholesterolemia, were induced by high fat diet. The insulin sensitivity was estimated with insulin tolerance test(ITT) and glucose tolerance test(GTT); the levels of blood glucose and total cholesterol(TC), and the activities of α-glucosidase, protein tyrosine phosphatase 1B(PTP1B), and fructose phosphate amide transferase(GFAT)were measured with biochemical methods, respectively. The sample H13(h) extracted from Rhodiola crenulata, Y12(y) from Cordyceps militaris, and D(d) from Rheum palmatum were selected according to the inhibition activity on both PTP1B and α-glucosidase in vitro, regulation on hypercholesterolemia in IRF mice, and effects on GFAT activity, respectively; their synergistic effects on the treatment of metabolic syndrome were determined in IRF mice; composition proportion of h∶y∶d was measured in accordance with the results of L8(27) orthogonal experiments targeting on the inhibition of both PTP1B and α-glucosidase; finally, the effective dose was assessed based on the effects on IGT and hypercholesterolemia, respectively, in IRF mice. In conclusion, the prescription JD is composed by R. crenulata, C. militaris, and R. palmatum with the rate of 20∶1∶1, and its effective oral dose is 200 mg•kg⁻¹ for treatment of metabolic syndrome; its main mechanism is to inhibit the targets PTP1B and α-glucosidase. Monarch drug, R. crenulata, can clear away the lung-heat, tonify Qi, resolve stasis and nourish the heart. Adjuvant drug, C. militaris, can tonify the lung Qi and the kidney essence, strengthen waist and knee, accompanied with R. crenulata to enhance the function of invigorating lung and kidney. Assistant drug, rhubarb, can clear heat, detoxify, and remove blood stasis. These three herbs are compatible to show the effects of tonifying Qi, nourishing essence, clearing heat, reducing phlegm and resolving masses for the treatment of metabolic syndrome.

Effect of different combined antihypertensive regimen on the erectile function in male hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To compare the effects of felodipine combined irbesartan regimen with that of felodipine combined metoprolol regimen on the sexual function in male hypertensive patients.</p><p><b>METHOD</b>One hundred and twenty-three male hypertensive patients (age 25 to 60) were randomly assigned to felodipine (5 mg/d) plus irbesartan (150 mg/d, n = 64) group and felodipine (5 mg/d) plus metoprolol (47.5 mg/d, n = 59) group. Dosage of felodipine were doubled after 4 weeks if the blood pressure were > or = 140/ 90 mm Hg (1 mm Hg = 0.133 kPa). At the baseline and post 24th week treatment, sexual function of patients was assessed by the International Index of Erectile Function (IIEF) Questionaire. Serum testosterone (T), sex hormone binding globulin (SHBG), 4-hydroxynonenal (HNE), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and Malonaldehyde (MDA) were measured by Radioimmunoassay (RIA), ELISA and TBA respectively.</p><p><b>RESULTS</b>Total prevalence of erectile dysfunction (ED), T, SHBG and HNE were similar between pre- and post-treatment in two groups (P > 0.05). On the other hand, the scores of the mild ED and sexual desire (SD) were improved and both serum 8-OHdG and MDA in patients with ED decreased [(146.02 +/- 60.54) ng/L vs. (139.89 +/- 62.03) ng/L, P = 0.048 and (6.59 +/- 1.75) micromol/L vs. (5.51 +/- 1.65) micromol/L, P = 0.039] in Felodipine plus Irbesartan group.</p><p><b>CONCLUSION</b>The results suggested that Felodipine + Irbesartan regimen may be superior to Felodipine + metoprolol regimen for male hypertensive patients with mild ED.</p>

Effect between felodipine plus irbesartan and felodipine plus metoprolol regimen on the sexual function in young and middle-aged women with hypertension / 中华心血管病杂志

<p><b>OBJECTIVE</b>To compare the effects between felodipine plus irbesartan and felodipine plus metoprolol regimen on blood pressure and the sexual function in young and middle-aged hypertensive women.</p><p><b>METHODS</b>In this prospective, randomized, parallelized, controlled and fixed combined therapy trial, 99 female patients (aged 18 to 60) with grade 1 and grade 2 hypertension (BP ≥ 140/90 mm Hg and < 179/109 mm Hg, 1 mm Hg = 0.133 kPa) were assigned to felodipine 5 mg q.d + irbesartan 150 mg q.d (F + I group, n = 49) and felodipine 5 mg q.d + metoprolol 47.5 mg q.d (F + M group, n = 50) group. Target blood pressure was < 140/90 mm Hg. The female sexual function index (FSFI) questionnaire, levels of serum estradiol and testosterone were assessed. Female sexual dysfunction was defined as a FSFI score of less than 25.5. Patients were followed up for 24 weeks.</p><p><b>RESULTS</b>The rate of achieving blood pressure goal between 2 groups was similar at the 4th, 8th, 12th and 24th weeks respectively (42.9% vs. 62.0% at 4th week, 89.8% vs. 90.0% at 8th week, 93.9% vs. 94.0% at 12th week, 98.0% vs. 96.0% at 24th week, P > 0.05). Compared to baseline, scores for the items related to "desire" and "arousal" were significantly improved (P < 0.05), the level of the serum estradiol was significantly elevated [(50.3 ± 37.4) pg/L vs. (54.4 ± 10.8) pg/L before menopause, (18.4 ± 2.9) pg/L vs. (20.2 ± 3.1)pg/L after menopause, P < 0.05] and the level of the serum testosterone was significantly decreased [(722.8 ± 277.1) ng/L vs. (650.0 ± 156.0) ng/L before menopause, (841.2 ± 279.3) ng/L vs. (761.9 ± 197.8) ng/L after menopause, P < 0.05] in the F + I group, while scores for the items related to "sexual desire" and "lubrication" were statistically reduced (P < 0.01), the concentration of the serum estradiol was significantly reduced [(57.4 ± 9.7) pg/L vs. (51.1 ± 12.1) pg/L before menopause, (19.8 ± 2.3) pg/L vs. (17.8 ± 3.3) pg/L after menopause, P < 0.01] and the level of the serum testosterone was significantly increased [(775.6 ± 217.8) ng/L vs. (886.0 ± 186.4) ng/L before menopause, (812.5 ± 311.3) ng/L vs. (914.4 ± 300.2) ng/L after menopause, P < 0.01] in the F + M group. FSFI score was negatively correlated with age and systolic blood pressure levels.</p><p><b>CONCLUSION</b>felodipine plus irbesartan or metoprolol for 24 weeks equally reduced blood pressure and the former regimen is superior to the latter on sexual function improvement in this patient cohort.</p>

Change of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases before and after percutaneous coronary intervention / 中南大学学报(医学版)

OBJECTIVE@#To investigate the culture of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases (CHD) before and after percutaneous coronary intervention (PCI), and to observe the cells shape and determine the cell number and proliferation activity.@*METHODS@#Ninety-five patients were divided into a CHD group(n=65) and a control group (n=30). The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF.EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colony-forming units(CFU) was counted by phase-contrast microscope.@*RESULTS@#The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. How-ever, the number of CFUs did not change in patients before and after the PCI.@*CONCLUSION@#PCI can increase endothelial progenitor cells in patients after the PCI; but 4 days after the PCI, this increase will not exist.

Effect of melatonin on the proliferation, apoptosis, and expression of bcl-2 in oxidized low-density lipoprotein-induced endothelial progenitor cells / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of melatonin(Mel) on the proliferation, apoptosis and expression of bcl-2 in oxidized low-density lipoprotein(ox-LDL)-induced endothelial progenitor cells (EPC) from human umbilical cord blood in vitro.@*METHODS@#Total mononuclear cells were isolated from human umbilical cord blood in vitro by Ficoll density gradient centrifugation, and the cells were plated on fibronectin-coated culture dishes. After 7 days, the attached cells were divided into 7 groups: a control group (normal cells), 3 ox-LDL groups[the attached cells were incubated with different concentrations of ox-LDL(5,10,and 20mg/L) for 24 hours], and 3 Mel groups[the attached cells were incubated with different concentrations of Mel (0.5,1.0, and 2.0 mmol/L) respectively for 24 hours before incubation with 10 mg/L ox-LDL]. EPC was identified by examining the expression of CD34, vascular endothelial growth factor receptor-2(VEGFR-2) and CD133 under a laser scanning confocal microscope. We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the effect of Mel and ox-LDL on the multiplication ability of EPC. Flow cytometry was used to detect the apoptosis. The expressions of Bcl-2 mRNA and protein were detected respectively by RT-PCR and immunohistochemistry technology.@*RESULTS@#After being exposed to the ox-LDL, the proliferation of EPC in the 3 ox-LDL groups was lower, and the apoptosis rate was higher than that in the control group in a dose-dependent manner (P<0.01); Mel was added at different concentrations before the ox-LDL incubation, and the cells in the 3 Mel groups showed higher proliferation and lower apoptosis rate than those of the 3 ox-LDL groups (P<0.01). Expression of Bcl-2 mRNA and protein of EPC in the 3 Mel groups was higher than that in the 3 ox-LDL groups (P<0.01).@*CONCLUSION@#Ox-LDL can inhibit the proliferation of EPC and promote the apoptosis of the cells by down-regulating the bcl-2 expression. Mel can inhibit these effects of ox-LDL.