Effects of different carbon sources on growth and active component contents in Salvia miltiorrhiza and S. castanea f. tomentosa hairy roots / 中国中药杂志

Salvia miltiorrhiza(Sm) and Salvia castanea f. tomentosa(Sc) hairy roots were used as experimental materials to study the effects of six different carbon sources, galactose, fructose, lactose, glucose, arabinose and sucrose(control), on fresh weight, dry weight, contents and yields of salvianolic acids and tanshinones. The results showed that galactose was most beneficial to the growth of two kinds of hairy roots, while lactose and arabinose were not conducive to their growth. As for Sm hairy roots, fructose significantly promoted the accumulation of salvianolic acid B, and the content increased by 5.801 times and 10.151 times compared with the control group, respectively. Glucose significantly promoted the accumulation of salvianolic acids. The content and yield of rosmarinic acid were 7.674 times and 9.260 times of that of the control group, and the content and yield of salvianolic acid B were 5.532 times and 6.675 times of the control group. For the hairy roots of Sc, galactose significantly increased the content and yield of rosmarinic acid, reaching 7.820 times and 9.944 times of the control group, respectively. Fructose promoted the increase of the content and yield of cryptotanshinone, reaching 9.242 times and 6.609 times of the control group, respectively. The study confirmed the optimal carbon source for the hairy root culture of Sm and Sc, and provided theoretical guidance for large-scale production of Sm drug-derived components and the utilization of Sc.

Analysis of HFE and Non-HFE Mutations in a Tibet Cohort with Iron Overload / 中国实验血液学杂志

OBJECTIVE@#The explore the molecular basis of iron-overload in Tibet nationality population of Tibet.@*METHODS@#The inpatients with iron-overload in our department from Dec. 1st 2014 to Jul.31st 2016 were enrolled in this study. Abdominal MRI and the mutation sites C282Y and H63D in HFE exon were examined. For HFE mutation-negative patients, the non-HFE mutation was detected, including 5 HJV mutations of G320V, p.Q312X, p.D249H, p.I281T, p.C321X and 2 TFR2 mutations: (Y250X, I238M), and 2 SLC40A1 mutations: (V162del, N144H).@*RESULTS@#Among 113 iron overload patients, only one showed homozygous p.H63D mutation, and one showed heterozygosis p.H63D mutation. In 73 patients accepted non-HFE gene detection, only one was heterozygosis p.D249N mutation in HJV, and one was heterozygosis p.I238M mutation in TFR2.@*CONCLUSION@#Currently, the pathogenic gene for Tibetan iron-overload has not yet been found.

Exogenous Nkx2-5 gene expression induces the expression of cardiac markers during P19 cell differentiation in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of homeobox gene Nkx2-5 in cardiac myogenesis.</p><p><b>METHODS</b>Two P19 cell lines, namely cells transfected with exogenous expression of Nkx2-5 gene and non-transfected cells, were cultured in suspension for 4 days to induce cell aggregation, and the cell aggregates were transferred to the Petri dish for further adherent culture. On days 4, 8, 12 and 16 of adherent culture, the expressions of α-sarcomeric actin (α-SA) and cardiac troponin T (cTnT) protein were detected by immunocytochemistry, and the mRNA expressions of GATA-4, α-myosin heavy chain (α-MHC) and atrial natriuretic factor (ANF) genes by RT-PCR.</p><p><b>RESULTS</b>In the transfected cells, α-SA and cTnT protein expressions were detected on days 8, 12 and 16 of adhere culture, and their expressions increased gradually with time. α-SA and cTnT expression was significantly higher on day 16 than on day 8 of culture (P<0.01). RT-PCR analysis of the transfected cell showed the presence of GATA-4 expression on day 4 of adherent culture, and the expression increased on days 8 and 12 but decreased on day 16. ANF and α-MHC expressions were found on days 8, 12, and 16, increasing gradually over time and showing significant differences from those on day 4 (P<0.05 or P<0.01). The expression of α-MHC was significantly higher on days 12 and 16 than on day 8 (P<0.05 or P<0.01), and ANF expression was significantly higher on day 16 than on days 8 and 12 (P<0.01). The non-transfected cells were negative for the expressions of all these genes.</p><p><b>CONCLUSION</b>Exogenous expression of Nkx2-5 gene can induce P19 cells to express cardiac markers in vitro.</p>

Diet-induced hyperhomocysteinemia exacerbates vascular reverse remodeling of balloon-injured arteries in rat / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>While hyperhomocysteinemia is associated with an increased risk of cardiovascular diseases, the effect of hyperhomocysteinemia on the vascular adventitia and vessel remodeling has not been clearly demonstrated. We investigated the effect of the hyperhomocysteinemia on adventitial hyperplasia and vascular remodeling following balloon injury in rats and the underlying mechanisms.</p><p><b>METHODS</b>Rats were fed with diet containing methionine for 4 weeks to increase plasma homocysteine before balloon injury. Vascular geometrical changes were assessed at different time points following balloon injury. The collagen deposition was determined by picrosirius red staining and immunohistochemical staining.</p><p><b>RESULTS</b>When compared with normal diet group, moderate hyperhomocysteinemia in methionine diet group significantly exacerbated adventitial hyperplasia at day 7 and collagen deposition mainly in the adventitia at day 28 following balloon injury. The increased plasma homocysteine level significantly increased collagen deposition in the adventitia. There was a negative correlation (r = -0.698; P < 0.01) between the luminal area and the collagen content in the adventitia on day 28 following balloon injury. In cultured adventitial fibroblasts isolated from rat aorta, 100 micromol/L L-homocysteine (L-Hcy) significantly down-regulated matrix metalloproteinase-2 activity by 43% as determined by in vitro gelatin zymography (P < 0.05) and up-regulated the expression of collagen type I by 187% (P < 0.05) assessed by Western blotting.</p><p><b>CONCLUSIONS</b>Hyperhomocysteinemia exacerbated vascular constrictive remodeling by accelerated neointima formation and collagen accumulation in the adventitia. Increased collagen deposition may be the underlying mechanism.</p>

The in vivo and in vitro effects on neovascularization of VEGF gene transfer with a new pseudotyped rAAV1 vector / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of vascular endothelial growth factor (VEGF) gene transfer with a new pseudotyped recombinant adeno-associated virus (rAAV) vector with AAV serotype 1 capsid protein (rAAV1) vector on neovascularization.</p><p><b>METHODS</b>PBS, rAAV1-GFP and rAAV1-VEGF vectors were added in C2C12 derived myotubes [10(5) vg (vector genomes) per cell]. Transfer efficiency was determined by fluorescent microscope and VEGF protein concentration in the culture media measured by ELISA. Ten days following ischemia in a hindlimb ischemic mouse model, PBS, 3 x 10(11)vg rAAV1-LacZ vectors and rAAV1-VEGF165 vectors were injected in ischemic thigh muscles. VEGF protein at ischemic thigh muscle was measured by ELISA at 1 month after vector infection. Capillaries and arterioles were observed by immunohistochemical analysis at 6 weeks after vector infection.</p><p><b>RESULTS</b>GFP expression was found in 60% - 80% myotubes at 120 hours after rAAV1-GFP infection. VEGF protein peaked at the 3rd day post rAAV1-VEGF infection with an average concentration of (567.7 +/- 16.8) pg/ml. Transfer efficacy in ischemic thigh muscle was 100% one month post rAAV1-LacZ infection. The average concentration of VEGF protein in ischemic skeletal muscles is (205.4 +/- 36.1) pg/mg total protein in rAAV1-VEGF165 treated mice. Extensive angiogenesis [(147.0 +/- 13.3)/mm(2)] and arteriogenesis [(17.0 +/- 1.2)/mm(2)] were observed in ischemic skeletal muscles at 6 weeks post rAAV1-VEGF165 injection.</p><p><b>CONCLUSION</b>Gene transfer with the new pseudotyped rAAV1-VEGF165 vector might be an effective therapeutic approach for ischemic cardiovascular diseases.</p>

Hyperplasia suppressor gene expression in vascular smooth muscle cells derived from normotensive and hypertensive patients underwent bypass surgery / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the novel hyperplasia suppressor gene (HSG) expression in vascular smooth muscle cells derived from normotensive and hypertensive patients underwent bypass surgery.</p><p><b>METHODS</b>Coronary heart disease patients underwent coronary artery bypass graft (CABG) operation in BEIJING ANZHEN hospital from 4 - 9, 2006 were enrolled in this study and divided into hypertensive group (n = 28) and normotensive group (n = 26). The preoperative venous blood samples were taken for serum biochemical and vasoactive peptides measurements. Total RNA was extracted from WBC, explanted-vessels and cultured VSMCs using TRIZOL and HSG expression was determined by Semi-Quantitative RT-PCR.</p><p><b>RESULTS</b>Body mass index (BMI) was significantly higher in hypertensive group compared to normotensive group (P < 0.01) while other biochemic parameters and vasoactive peptides were similar between the groups. BMI and GLU, BMI and SBP, BMI and DBP, GLU and TG, SBP and DBP were positively correlated (all P < 0.05). HSG expression in WBC, VSMCs and vessel tissue were significantly lower in hypertensive group than those in normotensive group (all P < 0.05). HSG expression in tissue was negatively correlated to BMI, SBP and DBP (all P < 0.05).</p><p><b>CONCLUSIONS</b>Reduced HSG expression and the negative correlation on vascular tissue HSG expression to BMI, SBP and DBP suggested a possible inhibitory role of HSG on VSMC proliferation and blood pressure.</p>

Recombinant adeno-associated viral vector 2-mediated gene transfer of VEGF165 improved porcine chronic myocardial ischemia / 中华心血管病杂志

<p><b>OBJECTIVE</b>Vectors commonly used for therapeutic angiogenesis such as adenovirus and plasmid had their own limitations. Adenovirus-associated virus (AAV) is a relatively new but probably more ideal vector as it is safe and efficient. We will study the efficiency of recombinant AAV-2 mediated vascular endothelial growth factor165 gene transfer in inducing angiogenesis and arteriogenesis, in improving blood flow and myocardium function in a porcine chronic myocardial ischemic model.</p><p><b>METHODS</b>Chinese experimental minipigs underwent placement of a left circumflex artery aneroid constrictor. Five weeks later, electrocardiogram, coronary angiography and magnetic resonance imaging were performed to confirm occlusion of LCX or ischemia of myocardium in LCX territory. Coronary blood flow, myocardium perfusion and left ventricular wall function were also evaluated. Then the animals were randomized to treatment with rAAV2-VEGF(165) (1 x 10(12) virus genome) or administration of PBS, both by direct myocardial injection. Three and six months after therapy, the animals were evaluated with regard to expression of VEGF(165) Capillary density and arteriole density of the ischemic myocardium, coronary angiography, myocardial perfusion and left ventricular function were also assessed six months after therapy.</p><p><b>RESULTS</b>Five weeks after aneroid occluder implantation, all the animals demonstrated complete or nearly complete occlusion of LCX and perfusion deficiency in LCX territory. Three months after therapy, expression of VEGF(165) mRNA and protein were higher in the VEGF than control group. The difference between the two groups diminished after six months. There was significant increase in capillary density (1404.06 +/- 250.48/mm(2) vs 976.88 +/- 344.79/mm(2), P < 0.05) and arteriole density (167.81 +/- 36.29/mm(2) vs 116.56 +/- 34.48/mm(2), P < 0.05) in VEGF group compared with control. Comparison of myocardial perfusion demonstrated marked differences between the two groups with significant improvement in animals treated with rAAV2-VEGF(165). No significant improvement in left ventricular function was seen in either the VEGF or control group.</p><p><b>CONCLUSIONS</b>Transmyocardial delivery of rAAV2-VEGF(165) resulted in VEGF gene expression for at least three months and stimulated angiogenesis and arteriogenesis in porcine model of chronic myocardial ischemia. Myocardial perfusion was also improved after VEGF gene delivery.</p>

Study on the frequency of antibiotics use per day among inpatients in 151 hospitals in 2003 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the antibiotics use in inpatients with hospitals under different scales.</p><p><b>METHODS</b>On the day of August 7, 2003, antibiotics use among inpatients were investigated, medical order to each inpatient was checked and forms were filled.</p><p><b>RESULTS</b>Data showed that 54.86% of the 89,539 inpatients from 151 hospitals were using antibiotics, out of which 48.98% were used for therapeutic purposes while 35.44% for prophylaxis, and 15.58% for both therapy and prophylaxis. Among those who had received antibiotics, 61.43% received one, 33.65% received two and 4.92% received 3 or more agents. In patients who were on antibiotics for therapy, only 23.92% of the samples were sent for pathogens detection. The prevalence of antibiotics use was different among different hospitals, with teaching hospitals having the lowest and hospitals with less than 300 beds were higher than those with more than 300 beds. Departments of respiratory diseases, intensive care unit, and pediatrics had the higher use of antibiotics.</p><p><b>CONCLUSION</b>Inpatients in 151 hospitals had high prevalence of antibiotics use, especially for prophylaxis use but pathogen detection was seldomly done. It is necessary to strengthen the rational use and management of antibiotics use in the hospitals.</p>

Effect of extract from Tripterygium hypoglaucum on proliferation and apoptosis of vascular smooth muscle cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of THW-4, an extract from Trypterygium hypoglaucum on proliferation and apoptosis of vascular smooth muscle cells (VSMC).</p><p><b>METHODS</b>VSMC derived from rabbit aorta were cultured in vitro and different concentrations of THW-4 were added in experimental groups. Cell proliferation was evaluated by MTT and apoptosis by transmission electron microscopy (TEM), TUNEL assay and Annexin V-FITC/PI double labelled assay.</p><p><b>RESULTS</b>The inhibitory effects of THW-4 on proliferation of VSMC displayed dose-time dependently, with the IC50 value of 15.6 microg/L at 48 hrs. Incubated with THW-4 (10-100 microg/L) for 56 hrs, VSMC mainly appeared early stage apoptosis and the percentage of apoptosis was found to raise along with the increase of the THW-4 concentration. Typical images of apoptosis could be observed under TEM and TUNEL assay showed increase of DNA segments with karyorrhexis and pyknosis after THW-4 treatment for 72 hrs. Analysis of cell cycle indicated the THW-4 mainly lead to the blockage of VSMC in G2/M stage.</p><p><b>CONCLUSION</b>THW-4 could inhibit the proliferation and induce the apoptosis of VSMC in vitro, suggesting that THW-4 is a potential agent for prevention of restenosis following angioplasty.</p>