血清内脂素水平与糖尿病肾病的关系研究分析

目的:研究空腹血清内脂素(Visfatin)水平,及与空腹血糖(FPG)、空腹胰岛素水平(FINS)等生化指标间的相关性,发现其在糖尿病肾病发生发展中的作用。方法:采用酶联免疫吸附法测定对照组(30例)、初发糖尿病组(45例)、糖尿病肾病组(68例)患者血清Visfatin水平及相关生化指标,应用稳态模型计算胰岛素抵抗指数(HOMA-IR)。结果:1初发糖尿病组、糖尿病肾病组的空腹血清内脂素水平显著高于对照组水平(P<0.01);糖尿病肾病组空腹血清内脂素水平高于初发糖尿病组水平(P<0.01);2血清内脂素与FPG、FINS、HOMA-IR、HbA1c水平呈正相关(r=0.353、0.503、0.540、0.403,P<0.05);3以血清内脂素为应变量,进行多元线性回归分析,糖化血红蛋白(β=0.197,P=0.002)、胰岛素抵抗指数(β=0.239,P=0.001)进入方程。结论:糖尿病肾病组空腹血清内脂素水平明显高于正常对照组和初发糖尿病组;糖化血红蛋白、胰岛素抵抗指数均是影响血清内脂素含量的关键因素;血清内脂素可能预测2型糖尿病的血糖控制程度,血清内脂素与糖尿病肾病的发生发展可能存在有密切关系。

冠心病患者血浆HO-1、IL-6及内脂素水平与冠状动脉病变严重程度相关性研究

目的:探讨冠心病患者血浆血红素加氧酶-1(HO-1)、白细胞介素-6(IL-6)和内脂素水平与冠状动脉病变严重程度的相关性。方法:选择胸痛患者217例,依据冠脉造影结果分为单支病变组、双支病变组、多支病变组和冠脉正常组;采用Gensini积分评价冠状动脉病变严重程度,应用酶联免疫吸附法测定血浆HO-1、IL-6及内脂素。结果:血浆HO-1水平冠脉病变组较正常组显著降低,并且三支血管病变组较双支血管病变及单支血管病变组显著降低(P<0.05);而血浆IL-6及内脂素水平冠脉病变组较正常组显著升高,并且三支血管病变组较双支血管病变及单支血管病变组显著升高(P<0.05)。Gensini积分≥30分组血浆HO-1水平明显低于<30分组(P<0.05),而IL-6及内脂素水平则明显升高(P<0.05)。相关分析显示:Gensini积分分别与IL-6及内脂素正相关(r=0.583,P<0.01;r=0.724,P<0.01),与血浆HO-1水平负相关(r=-0.517,P<0.01);HO-1分别与IL-6及内脂素负相关(r=-0.614,P<0.01;r=-0.517,P<0.01),而IL-6与内脂素正相关(r=0.648,P<0.01)。结论:血浆HO-1、IL-6及内脂素参与冠心病的发病过程,并与冠状动脉病变严重程度密切相关。

Responses of Membrane Lipid Peroxidation and Endogenous Hormones of Soybean Seedlings to UV-B Radiation and Rare Earth

[ Objective] The aim was to provide strategies for development of rare earth and control of environmental pollution. [ Method] Responses of membrane lipid peroxidation and endogenous hormones of soybean seedlings to UV-B radiation and rare earth were studied through hydroponics in laboratory. [ Result] The results showed that under irradiation of UV-B（ T1-0.15 W/m^2 and T2-0.45 W/m^2）, chlorophyll and indole-3-acetic acid（IAA） contents firstly decreased during the stress phase （1 -5 d） and then increased during the restoration phase （6 -9 d） while contents of malonadialdehyde（MDA） and abscisic acid（ABA） gradually increased during the imposition of UV-B radiation （1 -5 d） and subsequently decreased during recovery from UV-B stress （6 -9 d） . With adding of La （III） with the concentration of 20 mg · L^-1 , the decline/dse trend of chlorophyll, IAA, MDA and ABA contents was slowed down during the stress period while the rise/decline speed was accelerated during the recovery period. [ Conclusion] It suggests that the regulation of La （ III ） on membrane lipid peroxidation and endogenous hormones could increase chlorophyll and IAA contents, improve the metabolism of reactive oxygen species （ ROS）, inhibit membrane lipid peroxidation, decrease the accumulation amount of ABA and alleviate injury of UV-B radiation to soybean seedlings. Further, the protective potential of La （ III ） was better under low UV-B radiation than under high one.

Developmental Changes of the FAS and HSL mRNA Expression and Their Effects on the Content of Intramuscular Fat in Kazak and Xinjiang Sheep

Twenty-four male Kazak sheep and 30 Xinjiang fine wool sheep at different ages were selected to investigate the development-dependent expression levels of fatty acid synthase （FAS） gene and hormone-sensitive lipase （HSL） gene in muscle and their effects on the contents of intramuscular fat （IMF）. Longissimus dorsal muscle was sampled to measure IMF and total RNA was extracted to determine FAS and HSL mRNA expression levels by real-time PCR. The results showed that： l） The IMF content increased continuously with growth and showed significant differences （P 〈 0.05） between different age groups in male Kazak sheep, but in Xinjiang fine wool sheep there was no such difference observed. Furthermore, the IMF contents in Kazak were much higher （P 〈 0.01） than that of the other breed from day 30 to 90. 2） FAS mRNA expression level was the highest （P 〈 0.05） on day 0 in Kazak sheep and then declined with growth, in the other breed the gene showed a d‘ecline-rise-decline-rise＇ expression manner as the animals grew. HSL mRNA expression level had a similar model in two breeds, in Kazak sheep it was the highest on day 0 （P 〈 0.05） and in Xinjiang fine wool sheep on day 30 （P 〈 0.01）, then both decreased after this term. 3） In male Kazak sheep, FAS and HSL mRNA expression level were both negatively related to IMF content （r= -0.485 （P = 0.02）, r= -0.423 （P = 0.05））, and the ratio of FAS/HSL expression exhibited significantly negatively related IMF contents. In male Xinjiang sheep, there were no obvious relationship between FAS and HSL expression and IMF content （P 〉 0.05）.

Multiple pathogenic factor-induced complications of cirrhosis in rats: A new model of hepatopulmonary syndrome with intestinal endotoxemia

AIM： To develop and characterize a practical model of Hepatopulmonary syndrome. （HPS） in rats.METHODS： The experimental animals were randomized into five feeding groups： （1） control （fed standard diet）, （2） control plus intraperitoneal injection with lipopolysaccharide （LPS）, （3） cirrhosis （fed a diet of maize flour, lard, cholesterol, and alcohol plus subcutaneously injection with carbon tetrachloride （CCI4） oil solution）, （4） cirrhosis plus LPS, and （5） cirrhosis plus glycine and LPS. The blood, liver and lung tissues of rats were sampled for analysis and characterization. Technetium 99m-labeled macroaggregated albumin （Tc99m-MAA） was used to test the dilatation of pulmonary microvasculature.RESULTS： Typical cirrhosis and subsequent hepato- pulmonary syndrome was observed in the cirrhosis groups after an 8 wk feeding period. In rats with cirrhosis, there were a decreased PaO2 and PaCO2 in arterial blood, markedly decreased arterial 02 content, a significantly increased alveolar to arterial oxygen gradient, an increased number of bacterial translocated within mesenteric lymph node, a significant higher level of LPS and tumor necrosis factor-α （TNF-α） in plasma, and a significant greater ratio of Tc99m-MAA brain-overlung radioactivity. After LPS administration in rats with cirrhosis, various pathological parameters got worse and pulmonary edema formed. The predisposition of glycine antagonized the effects of LPS and significantly alleviated various pathological alterations.

Innate immune reactivity of the liver in rats fed a choline-deficient L-amino-acid-defined diet

AIM： To investigate the innate immune reactivity of tumor necrosis factor-alpha （TNF-α）, Toll-like receptor 4 （TLR4）, and CD14 in the liver of non-alcoholic steatohepatitis （NASH） model rats. METHODS： Male F344 rats were fed a cholinedeficient L-amino-acid-defined （CDAA） diet. The rats were killed after 4 or 8 wk of the diet, and their livers were removed for immunohistochemical investigation and RNA extraction. The liver specimens were immunostained for TNF-α, TLR4, and CD14. The gene expressions of TNF-α, TLR4, and CD14 were determined by reverse-transcriptase polymerase chain reaction （RT-PCR）. Kupffer cells were isolated from the liver by Percoll gradient centrifugation, and were then cultured to measure TNF-α production. RESULTS： The serum and liver levels of TNF-~ in the CDAA-fed rats increased significantly as compared with the control group, as did the immunohistochemical values and gene expressions of TNF-α, TLR4, and CD14 with the progression of steatohepatitis. TNF-α production from the isolated Kupffer cells of the CDAAfed rats was elevated by lipopolysaccharide stimulation. CONCLUSION： The expressions of TNF-α, TLR4, and CD14 increased in the NASH model, suggesting that

Visceral fat and insulin resistance as predictors of non-alcoholic steatohepatitis

AIM： To examine whether visceral fat is associated with non-alcoholic steatohepatitis （NASH）, to assess for parameters associated with visceral adiposity and to investigate for factors associated with fibrotic severity in NASH. METHODS： Thirty NASH and 30 control subjects underwent biochemical tests, anthropometric assessment, bioelectrical impedance, dual energy X-ray absorptiometry and abdominal fat study by CT scan. Liver biopsies were graded according to the Brunt criteria. RESULTS： NASH subjects had elevated blood pressure, body mass index, waist circumference and waist-to-hip ratio. A greater number of diabetes rnellitus, impaired glucose tolerance test and HOMA-IR 〉 3.5 were found in NASH patients. HOMA-IR 〉 2.8 （OR 20.98, 95% CI 3.22-136.62; P 〈 0.001） and visceral fat area 〉 158 cm^2 （OR 18.55, 95% CI 1.60-214.67; P = 0.019） were independent predictors for NASH. Advanced stage of NASH was found in 15 （50%） patients. HOMA-IR 〉 3.5 （OR 23.12, 95% CI 2.00-266.23; P = 0.012） and grading of portal inflammation （OR 7.15, 95% CI 1.63-31.20; P = 0.009） were determined as independent risk factors for advanced stage of NASH. CONCLUSION： Obesity （especially central obesity） and metabolic syndrome are common in Thai NASH. Insulin resistance and elevated visceral fat are risk factors for the presence of NASH. The advanced stage of thedisease is related to insulin resistance.

Endocytosis of adiponectin receptor I through a clathrin- and Rab5-dependent pathway

In eukaryotic cells, receptor endocytosis is a key event regulating signaling transduction. Adiponectin receptors belong to a new receptor family that is distinct from G-protein-coupled receptors and has critical roles in the pathogenesis of diabetes and metabolic syndrome. Here, we analyzed the endocytosis of adiponectin and adiponectin receptor 1 （AdipoR1） and found that they are both internalized into transferrin-positive compartments that follow similar traffic routes. Blocking clathrin-mediated endocytosis by expressing Eps15 mutants or depleting K^＋ trapped AdipoR1 at the plasma membrane, and K^＋ depletion abolished adiponectin internalization, indicating that the endocytosis of AdipoR1 and adiponectin is clathrin-dependent. Depletion of K^＋ and overexpression of Eps15 mutants enhance adiponectin- stimulated AMP-activated protein kinase phosphorylation, suggesting that the endocytosis of AdipoR1 might down-regulate adiponectin signaling. In addition, AdipoR1 colocalizes with the small GTPase Rab5, and a dominant negative Rab5 abrogates AdipoR1 endocytosis. These data indicate that AdipoR1 is internalized through a clathrin- and Rab5- dependent pathway and that endocytosis may play a role in the regulation of adiponectin signaling.

Effect of JIANPI HUOXUE decoction on inflammatory cytokine secretion pathway in rat liver with lipopolysaccharide challenge

AIM： To evaluate the effect of Chinese traditional medicinal prescription, JIANPI HUOXUE decoction （JHD） on cytokine secretion pathway in rat liver induced by lipopolysaccharide （LPS）. METHODS： Twenty-four male SD rats were divided into normal group （n = 4）, model group （n = 10） and JHD group （n = 10） randomly. Rats in model group and JHD group were administrated with normal saline or JHD via gastrogavage respectively twice a day for 3 d. One hour after the last administration, rats were injected with LPS via tail vein, 50 μg/kg. Simultaneously, rats in normal group were injected with equivalent normal saline. After LPS stimulation for 1.5 h, serum and liver tissue were collected. Pathological change of liver tissues was observed through hematoxylineosin （H.E.） staining. Tumor necrosis factor alpha （TNF-α） in serum were assayed by enzyme linked immunosorbent assay （ELISA）. The protein expression of TNF-α, phosphorylated inhibit-κB （p-κB） and CD68 in liver were assayed by Western blot. The distribution of CD68 protein in liver was observed through immunohistochemical staining. The mRNA expression of TNF-α, interleukin-6 （IL-6）, CD14, toll-like receptor 2 （TLR2） and TLR4 in liver were assayed by real-time RT-PCR.RESULTS： Predominant microvesicular change, hepatocyte tumefaction and cytoplasm dilution were observed in liver tissues after LPS administration as well as obvious CD68 positive staining in hepatic sinusoidal. After LPS stimulation, serum TNF-α （31.35 ± 6.06 vs 12225.40 ± 9007.03, P 〈 0.05）, protein expression of CD68 （1.13 ± 0.49 vs 3.36 ±1.69, P 〈 0.05）, p-IκB （0.01 ±0.01 vs 2.07 ＋ 0.83, P 〈 0.01） and TNF-α （0.27 ± 0.13 vs 1.29 ± 0.37, P 〈 0.01） in liver and mRNA expression of TNF-α （1.96 ± 2.23 vs 21.45 ±6.00, P 〈 0.01）, IL-6 （4.80 ± 6.42 vs 193.50 ± 36.36, P 〈 0.01） and TLR2 （1.44 ± 0.62 vs 4.16 ± 0.08, P 〈 0.01） in liver were also increased significantly. These pathological changes were all improved in .1HD group. On the other hand, TLR4 mRNA （1.22 ± 0.30 vs 0.50 ± 0.15, P 〈 0.05） was down-regulated and CD14 mRNA increased but not significantly after LPS stimulation. CONCLUSION： JHD can inhibit cytokine secretion pathway induced by LPS in rat liver, which is probably associated with its regulation on CD68, p-IκB and endotoxin receptor TLR2.

The preparation of low electroendosmosis agarose and its physico-chemical property

Studies on Gelidium amansii agar fractionations were carried out in this paper. Gelidium amansii agar was fractionated on DEAE-Cellulose, and four fractions were obtained sequentially. The fractions were analyzed on physical and chemical properties, and IR and 13C-NMR spectroscopy applied for elucidating the chemical structure. Among the four fractions obtained, water fraction measured up to the standard of low EEO agarose. The sulfate content, ash content, electroendosmosis and gel strength (1%) of water fraction were 0.16%, 0.34%, 0.12 and 1 130g/cm2 respectively, similar to those of the Sigma products.

Effects of the heparin-coated Wiktor-I stent on recovering the balance of ET-1 and CGRP after intracoronary stent implantation

Objective: To study the effects of heparin-coated stent (Wiktor-I) on recovering the balance between endothelin- 1(ET- 1) and calcitonin gene-related peptide (CGRP) both in the plasma of coronary sinus and peripheral vein after stent implantation. Methods: The patients with coronary artery disease (n=30) were divided into non-coated stent (NCS) group (n=16) and heparin-coated stent (HCS) group (n=14). Circulating ET-1 and CGRP levels were measured by radioimmunoassay (RIA) in the 2 groups at the paired sampling sites, namely the coronary sinus (Cs) and peripheral vein (Pv). The ratio of ET/CGRP was also calculated to reflect the balance of ET and CORP. Results: In NCS group plasma ET-1 concentrations exhibited 2 peaks at 0 mm and 12 h after stent implantation, CORP levels were highly consistent with that of ET-1, and the ratio of ET/CGRP increased markedly at post-PTCA 5 mm, persisting until post-stent 12 h. While in HCS group ET-1 levels and the ratio of ET/CGRP were decreased dramatically compared with that of NCS group at 5 min,10 mm and 12 h post-stent (P＜0.05, P＜0.01 and P＜0.05, respectively＝. There was no remarkable difference of CGRP levels between the 2 groups. Conclusions: Heparin-coated stent can decrease the post-stent circulating ET-1 levels and recover the balance between ET-1 and CGRP.

MyD88/PI3-K/NF-κB pathway mediates the antagonism of lipoxin A_4 on LPS-induced synthesis of interleukins in endothelial cells

In order to investigate whether lipoxin A4 （LXA4） has an antagonistic effect on lipopolysaccharide （LPS）-induced synthesis of interleukin （IL）-β3, IL-6 and IL-8 in rat pulmonary microvascular endothelial cells （PMVEC）, and to explore the molecular mechanisms of signal pathway in LXA4 actions, cultured PMVEC were treated with LPS, with or without preincubation with LXA4. Proteins of IL-β3, IL-6 and IL-8 in supernatant were analyzed by enzyme-linked immunosorbent assay （ELISA）. Expressions of mRNA of IL-β3, IL-6 and IL-8 were determined by RT-PCR. Expressions of phosphorylation of phosphoinositide 3-kinase （PI3-K） and myeloid differentiation factor 88 （MyD88） were analyzed by Western blot. Activities of DNA-binding of nuclear factor-kappa B （NF-κB） and activator protein-1 （AP-1） were measured by electrophoretic mobility shift assay （EMSA）. The results showed that LPS induced production of IL-β3, IL-6 and IL-8 in rat PMVEC via MyD88/PI3-K/NF-κB and AP-1 pathway-dependent signal transduction. LPS-stimulated expression of PI3-K, activities of NF-κB and AP-1, secretion of protein and expression of mRNA of IL-β3, IL-6 and IL-8 but not MyD88 expression in PMVEC were inhibited by LXA4 in a dose-dependent manner. In conclusion, LXA4 inhibits synthesis of IL-β3, IL-6 and IL-8 by down-regulation of PI3-K/NF-κB and AP-1 signal pathway in PMVEC.

EFFECT OF HEMORRHAGIC SHOCK ON ENDOTOXININDUCED TNF PRODUCTION AND ITS MOLECULAR MECHANISM IN RATS

The present study was designed to investigate the production of tumor necrosis factor a (TNFα) in-duced by low-dose (1μg/kg) lipopolysaccharide (LPS) and its cellular source after hemorrhagic shock(HS) in rats, and to further analyze the mechanism for increased sensitivity to LPS through looking at ex-pression of lipopolysaccharide -binding protein (LBP ) mRN A in t he liver, lungs and kidneys. lt wa s foundin uiuo that plasma TNFa levels in the HS+LPS group were 20-fold higher than that in the HS group (P<0. 01 ), and 2. 7-fold higher than that in the LPS group (P<0. 05). lt was shown in ndro that the ca-pacity of peripheral white blood cells to produce TNFa in response to LPS stimulation was significantly de-creased by 126 % (P<0. 01 ) and 57% (P<0. 05) compared with pre-shock levels and the sham group re-spectively at the end of resuscitation following shock, and was still markedly decreased 3 hours after resus-citation, while the capacity of Kupffer Cells was significantly increased by 110% compared with the shamgroup (P<0. 01) after shock and resuscitation. Results from RT-PCR showed that expression of LBPmRNA in the liver, lungs and kidneys was increased after shock and resuscitation. It is suggested thathemorrhagic shock could significantly enhance endotoxin-induced TNFa production, which might be due toup-regulation of LBP expression in tissues after shock, and tissue macrophages might be the main source ofcytokine production.

Influence of moxibustion temperatures on blood lipids, endothelin-1, and nitric oxide in hyperlipidemia patients

OBJECTIVE： To observe the influence of moxibustion temperature on blood lipids, endothelin-1（ET-1）, nitric oxide（NO）, and ET-1/NO in hyperlipidemia patients. METHODS： Forty-two primary hyperlipidemia patients were randomly divided into two groups of 21 and treated with moxibustion at different temperatures. Moxibustion was performed with the moxa roll 2.5-3.0 cm from the skin in the treatment group and 4 cm in the control group, 10 min per point, once every other day. Skin temperature was precisely measured with a thermometer during moxibustion. After a 12-week treatment, seven measurements of blood lipids, ET-1, and NO were recorded. RESULTS： Total cholesterol and triglyceride, were lower in the treatment group than in the control group（P0.05）. Serum ET-1 and ET-1/NO was obvi-ously lowered in the treatment group（P0.001）. Moxibustion regulated NO and ET-1/NO in the treatment group much better than in the control group. CONCLUSION： Moxibustion can regulate blood lipids and clear blood vessels. Moxibustion at 45℃has a better effect than moxibustion at 38℃ on regulating blood lipids and protecting vascular endothelial function, indicating that suitable temperature influences the curative effect of moxibustion.

Follicle-stimulating hormone increases the intramuscular fat content and expression of lipid biosynthesis genes in chicken breast muscle

Intramuscular fat （IMF） is a crucial factor in the quality of chicken meat. The genetic basis underlying it is complex. Follicle-stimulating hormone （FSH）, well-known as an effector in reproductive tissues, was recently discovered to stimulate abdominal fat accumulation in chicken. The effect of FSH on IMF accumulation and the underlying molecular regulatory mechanisms controlling both IMF and abdominal fat deposition in vivo are largely unknown. In this study, two groups of chickens were treated with chicken FSH or a placebo. The lipid content of breast muscle, abdominal fat volume, and serum concentrations of FSH were examined. Related genes implicated in breast muscle and abdominal fat accumulation were also investigated. Compared to the control group, the triglyceride （TG） content of breast muscle and the percentage of abdominal fat in FSH-treated chickens were significantly increased by 64.9% and 56.5% （P〈0.01）, respectively. The FSH content in the serum of FSH-treated chickens was 2.1 times than that of control chickens （P〈0.01）. Results from quantitative real-time polymerase chain reaction （qRT-PCR） assays showed that relative expression levels of fatty acid synthase （FAS）, lipoprotein lipase （LPL）, diacylglycerol acyltransferase 2 （DGAT2）, adipocyte fatty acid binding protein （A-FABP）, and peroxisome proliferator-activated receptor ~ （PPARy） were significantly upregulated in breast muscle following FSH treatment （P〈0.01）. Treatment with FSH also signifi- cantly increased relative expression levels of FAS, LPL, DGA T2, A-FABP, and PPARy in abdominal fat tissue （P〈0.05） The results of principal component analysis （PCA） for gene expression （breast muscle and abdominal fat） showed that the control and FSH treatment groups were well separated, which indicated the reliability of the data. This study demonstrates that FSH plays an important role in IMF accumulation in female chickens, which likely involves the regulation of biosynthesis genes related to lipid metabolism.

Spleen-Yang-deficiency patients with polycystic ovary syndrome have higher levels of visfatin

OBJECTIVE:To study serum visfatin levels in women with polycystic ovary syndrome(PCOS)grouped by Traditional Chinese Medicine(TCM)patterns.To study the correlations of serum visfatin levels with homeostatic model assessment insulin resistance(HOMA-IR),fasting plasma glucose(FPG),fasting insulin(FINS),body mass index(BMI),testosterone(T),total cholesterol(TC),and triglycerides(TG).METHODS:Two hundred and twelve PCOS patients were placed into the following TCM pattern subgroups:Kidney-Yang deficiency(KYD)group,Spleen-Yang deficiency(SYD)group,stagnant Liver-Qi transforming into heat(SLQTH)group,and Kidney-Yin deficiency(KYIND)group.The correlations between serum visfatin levels and HOMA-IR,FPG,FINS,BMI,T,TC,andTG were analyzed.RESULTS:Of all patients with PCOS,there were 82in the KYD group(38.6%),67 in the SYD group(31.6%),37 in the SLQTH group(17.5%),and 26 in the KYIND group(12.3%).Visfatin levels in all PCOS subgroups were higher than those in the control group(P<0.01 or P<0.05).Among these subgroups,the visfatin levels in the SYD group were significantly higher than those in the other three TCM pattern groups(P<0.05).There were no statistical differences among the remaining three pattern groups.The levels of BMI,FINS,HOMA-IR,T,and TG were significantly higher in all subgroups than those in the control group(P<0.05).There were no significant differences in FPG and TC between all PCOS subgroups and the control group(P>0.05).The SYD group had higher levels of FINS and HOMA-IR compared with the KYD,SLQTH,and KYIND groups(P<0.05).In all subgroups,after controlling for BMI,TG,TC,and age,visfatin was positively correlated with FINS(r=0.197,P=0.015)and HOMA-IR(r=0.173,P=0.033),and was not correlated with T.CONCLUSION:KYD and SYD patterns are most common in PCOS patients.Increased visfatin is a common pathophysiologic manifestation in PCOS patients.The SYD group had the highest levels of visfatin,and visfatin was positively correlated with FINS and HOMA-IR.

Detection of bacterial endotoxin in paclitaxel liposome

Based on current research, there are three technologies during the test of bacterial endotoxin of liposomes：（1） extraction of bacterial endotoxin from liposomes;（2） addition of bacterial endotoxin in the process of recovery test; and（3） elimination of the interference factors from drugs and excipients. In the present study, we pointed out that the key technologies to test bacterial endotoxin from paclitaxel liposome included following steps： extraction of bacterial endotoxins from ethanol-dissolved liposomes; preparation of positive control of recovery solution by adding 0.01 m L standard endotoxins in 1 m L liposome ethanol solution; and the use of 0.5% human albumin to eliminate the interference from detection, and accurate detection of the bacterial endotoxin of liposomes.

Effect of polydatin on phospholipase A_2 in lung tissues in rats with endotoxic shock

Objective: To study the effect of polydatin on p hospholipase A2 in lung tissues in rats with endotoxic shock. Methods: Thirty-two healthy male Wistar rats were employed in this study. A total of 8 rats received normal saline intravenously （control grou p）,8 rats received 10 mg/kg of endotoxin （endotoxic shock group）,8 rats re ceived 1 mg/kg of polydatin after endotoxin injection （polydatin treatment g roup）,and 8 rats received 1 mg/kg of polydatin （polydatin prevention group） 30 minutes before endotoxin injection. Mean arterial pressure was measured once hal f an hour. Lung tissues were collected 6 hours later. Phospholipase A2 activit y was measured with acid titration. The gene expression of secretory phospholipa se A2 type IIA was detected with reverse transcription polymerase chain reacti on. Meanwhile,the histological changes of the lungs among four groups were comp ared through microscopic examination.Results: Phospholipase A2 activity and the gene expression of secretory phospholipase A2 type IIA increased after endotoxin injection,but polydatin could inhibit these effects of endotoxin. Obvious morphological eviden ce could be found in the lung pathological sections and the protective effect of polydatin was most significant in the polydatin prevention group.Conclusions: Polydatin has prophylactic and therapeutic effects （the former is more distinct than the latter） on acutely injured lungs in rats with endotoxic shock and which suggests that polydatin may be a phospholipase A 2 inhibitor.

Adiponectin levels are associated with the number and activity of circulating endothelial progenitor cells in patients with coronary artery disease

Objective:To study the relationship between plasma adiponectin concentration and the functional activities of circulating endothelial progenitor cells(EPCs) in patients with coronary artery disease(CAD).Methods:Circulating EPCs were enumerated as AC133+/KDR+ cells via flow cytometry and identified by co-staining with DiI-acLDL and fluorescein isothiocy-anate(FITC)-conjugated lectin under a fluorescent microscope.The migratory capacity of EPCs was measured by modified Boyden chamber assay.Adhesion capacity was performed to count adherent cells after replating EPCs on six-well culture dishes coated with fibronectin.Results:The number of circulating EPCs(AC133+/KDR+ cells) decreased significantly in CAD patients,compared with control subjects [(74.2±12.3) vs(83.5±12.9) cells/ml blood,P<0.01].In addition,the number of EPCs also decreased in CAD patients after ex vivo cultivation [(54.4±8.6) vs(71.9±11.6) EPCs/field,P<0.01].Both circulating EPCs and differentiated EPCs were positively correlated with plasma adiponectin concentration.The functional activities of EPCs from CAD patients,such as migratory and adherent capacities,were also impaired,compared with control subjects,and positively correlated with plasma adiponectin concentration.Conclusion:The study demonstrates that the impairment of the number and functional activities of EPCs in CAD patients is correlated with their lower plasma adiponectin concentrations.

In vivo expression of lipopolysaccharide binding protein and its gene induced by endotoxin

Objective: To investigate the expression of lipopolysaccharide binding protein (LBP) and its gene in rats with endotoxemia and explore the role of LBP in the response of host to endotoxin. Methods: Thirty Wistar rats were divided randomly into five groups: the normal group and the endotoxemia groups (1, 3, 6, 12 hours after LPS injection, respectively). The level of plasma endotoxin was determined by the Limulus Amebocyte Lysate assay. The expression of LBP mRNA in hepatic tissue was examined by reverse transcription polymerase chain reaction (RT PCR). Plasma levels of LBP, tumor necrosis factor (TNF) α and interleukin (IL) 6 were measured by enzyme linked immunosorbent assay (ELISA). Morphologic changes of hepatic tissue were observed under transmission electron microscope. Results: The level of plasma endotoxin peaked at 1 h after LPS injection, then declined, but was still higher than that of the normal group at 12 h; intrahepatic expression of LBP mRNA and plasma LBP increased with time after LPS stimulation; TNF α and IL 6 in plasma increased with upregulation of LBP expression; there were significant differences between the normal group and endotoxemia groups (P< 0.05 ). Activation of Kupffer cells and injury of hepatocytes could be seen in rats with endotoxemia. Conclusions: LPS can upregulate the intrahepatic expression of LBP mRNA and increase the plasma LBP level. Under certain conditions, LBP may enhance the sensitivity of host to the toxic effects of LPS.