To explore the kinetics of LBP/CD14 system in patients with hemodialysis, and further to analyse its role in (he development of systemic inflammation response based on the changes of plasma endotaxin, TNF and IL-6 levels in patients with uremia treated with hemodilysis. Methods Sixteen patients with end-stage renal failure (8 cases with hemodialysis, 8 cses without hemodialysis) were selected for this study. limulus amebocyte lysate chromogenic assay, ELISA and cell in situ hybridization were used to determine the changes in endotoxin, LBP, TNF and IL-6 levels in plasma, and expression of CD14 mRNA in the monocytes. Results (l)Plasma LBP levels in patients with hemodialysis were significantly higher than those in patients without hemodialysis. The expression of CD14 mRNA in the monocytes in patients with hemodialysis was also more obvious. Both of them incresed much more in hemodialysis. (2)Plasma endotoxin levels in patients with hemodialysis, though being significantly higher than in patients without hemodialysis, were in low-level (61.7 ? 10.6 pg/ml). (3) Plasma TNF and IL-6 levels were markedly increased in patients with hemodialysis, which were significantly correlated with plasma LBP levels. Conclusion LBP/CD14 system in patients with hemodialysis is markedly up-regulated, which might be the important mechanism for low-level endotoxemia to exert its effects in hemodialysis.

BACKGROUND: The abnormality of the function of arcuate nucleus may be an important factor of obesity. It has been known that the mechanisms of acupuncture in treating obesity are related to nervous and neurohumoral regulation. What is the regulating effect of acupuncture on the function of arcuate nucleus?OBJECTIVE: To study the effects of acupuncture on the function of arcuate nucleus of obese rats, and further investigate central nervous functional mechanism of reducing weight by acupuncture.DESIGN: Randomized controlled study based on the experimental animals.SETTING: Acupuncture institute in second clinical medical college of a university of traditional Chinese medicine, and a population management college.MATERIALS: This experiment was carried out in the Acupuncture Institute of Second Clinical Medical College, Nanjing University of Traditional Chinese Medicine between April and October 2002. One-month old male SD rats just in ablactation were selected.METHODS: Rats fed with ordinary wholesome rat-feed were in the normal group. The successfully established experimental obese rats models were randomly divided as control group and acupuncture group with 12 rats in each group. Rats in the acupuncture group were given acupuncture treatment for 14 days, and rats in the normal and control groups were put into rat fixation-machine for 15 minutes every day, lasting for 14 days. Body mass, Lee' s index, body lipid, level of central and peripheral leptin and insulin(INS) as well as the frequency of spontaneous discharge of nerve cell in the arcuate nucleus(ARC) of hypothalamus in obese rats were observed with nervous electrophysiological and nervous biochemical technology before and after acupuncture.MAIN OUTCOME MEASURES: ① Effect of acupuncture on obesity index, fat contents of pericardium, kidney and epididymis of experimental obese rats. ② Effect of acupuncture on the frequency of spontaneous discharge of ARC nerve cell of hypothalamus in obese rats. ③ Effect of acupuncture on the contents of hypothalamic and serum leptin and INS of rats in each groups. ④ Correlation of the frequency of spontaneous discharge of ARC nerve cell of hypothalamus with the level of hypothalamic leptin and INS.RESULTS: The body mass[(517.74±29.35) g], Lee' s index (319.85 ± 3.96) and body fat content of obesity rats [ (13.88 ± 1.32) g] were all significantly higher than those in the normal rats, the differences were significant(P ＜ 0.01); the frequency of spontaneous discharge of nerve cell in ARC of hypothalamus in obese rats [ (3.12 ± 1.92) Hz] were all markedly lower than those of the normal rats[ (8.99 ± 2.71 ) Hz], the differences were significant (P ＜ 0.01 ); the frequency of spontaneous discharge of nerve cell in ARC was negatively correlated with body mass, Lee' s index and body lipid( r = -0. 592, -0. 672, -0. 521 ) . Acupuncture significantly increased the frequency of spontaneous discharge of nerve cell in ARC of hypothalamus in obese rats [(9.75 ±2.02) Hz] as compared with the control group, the differences were significant( P ＜ 0. 01 ).CONCLUSION: The good regulating effect of acupuncture on the function of arcuate nucleus of hypothalamus might be one of mechanisms in the antiobesity of acupuncture.

The duck circovirus (DuCV) infection in sick ducks from Fujian Province was investigated. The liver samples of 43 sick Muscovy ducks with infectious serositis were collected from 12 duck farms in Fujian Province.Based on the published sequences of DuCV, two primers were designed for the detection of DuCV and four pairs of primers were designed to amplify four overlapping fragments that cover the complete genome of DuCV. The specific PCR products were amplified from positive samples. The fragments were then cloned into pMD18-T vector and sequenced, and the full length genomic sequence of the FJ0601 isolate of DuCV was obtained. PCR analysis showed that the proportion of ducks which were positive for circovirus was 79% and 10 out of the 12 farms were positive. Sequence analysis showed that the complete genome of DuCV-FJ0601 was 1988 bp and possessed features common to the family Circoviridae which included a stem-loop structure and the Rep protein motifs. Homology analysis showed that FJ0601 isolate of DuCV had 97.3%～97.5% nucleotide sequence identity to all the four Taiwan isolates (TC1/2002, TC2/2002, TC3/2002, TC4/2002), 82.9% identity to the America (33753-52) isolate and 82.3% identity to the Germany isolate. Phylogenetic analysis with Clustal W, however,showed that FJ0601 isolate of DuCV was on a common branch with Taiwan isolates, and Germany and America isolates belonged to the other branch.

Objective:To investigate if acupuncture has an inhibitory effect on fatty toxicity and its possible mechanism. Methods: Thirty-four patients with simple obesity were clinically treated with acupuncture for 3 courses. Before and after treatment, body mass index(BMI) and fat percentage(F%) were determined, and serum insulin and adiponectin were measured by enzyme linked immunoassay, blood lipid and sugar by biochemical colorimetry and eight kinds of free fatty acids(FFAs) by high performance liquid chromatography. Twenty normal persons were selected as a control. Results:The total efficacy rate was 88.2%. Acupuncture could increase the decreased insulin sensitivity index(ISI) in the patients(t=-5.103, P=0.000). The correlation of a decrease in F% with an increase in unsaturated fatty acid after acupuncture was of significance(r=0.402, P=0.019) and its correlation with an increase in the ratio between unsaturated fatty acid and saturated fatty acid was also of significance(r=0.348, P=0.044). The correlation between increases in high density lipoprotein and in eicosapentaenoic acid was of significance(r=0.352, P＝0.041). The correlation between a decrease in BMI and an increase in high density lipoprotein was of significance(r=0.357, P=0.038). Adiponectin level significantly rose after acupuncture (compared with pre-treatment, P=0.000). The correlation between a decrease in FFAs and an increase in adiponectin was of significance(r=-0.349, P=0.043). Conclusion: Acupuncture can lower the levels of free fatty acids in the patients and increase their sensitivity to insulin to inhibit the fatty toxicity. The inhibitory effect of acupuncture on fatty toxicity is somewhat related to a rise in adiponectin level.

AIM:To investigate the protective effect of bactericidal/permeability-increasing protein (BPI) on sepsis induced by intra-abdominal infection in rats and its mechanism.METHODS:Intra-abdominal infection induced sepsis was reproduced by cecal ligation and puncture (CLP). BPI or equal volume of physiological saline was intra-abdominally given immediately after CLP and 12 hours after CLP respectively (2.5 mg/kg of BPI each time). Plasma endotoxin levels were determined with limulus amebocyte chromogenic assay.RESULTS:(1)The survival time in BPI group was significantly higher than in physiological saline (PS) group. (2)The values of MAP, LVSP, IP, d p /d t max and -d p /d t max in BPI group, although decreasing ,were markedly higher than those in PS group. (3) Plasma glutamate-pyruvate transaminase and urea nitrogen levels in BPI group, though increasing, were significantly lower than those in PS group.(4) There was no significant change of plasma endotoxin levels in BPI group, while plasma endotoxin levels were markedly increased in PS group. There was significantly different between two groups. CONCLUSIONS:BPI has an obvious protective effect on intra-abdominal infection induced sepsis, which might be related to its antagonism against endotoxin.

BACKGROUND:Both acupuncture and moxibustion and drug ale effective methods to treat simple obesity.Whether electroacupuncture is different from sibutramine to affect adipocyte products in obese organisms?OBJECTIVE:To observe the different effects of electroacupuncture and sibutramine on adipocyte produces in obesity rats and to investigate the mechanism of weight reduction by acupuncture and moxibustion.DESIGN:Layering-randomized controlled study.SETTING:Laboratory of Acupuncture and Pharmacology.Nanjing University of Chinese Medicine.MATERIALS:12045-day-old male SD rats were provided by Shanghai Experimental Animal Center.The experiment was in accordance with animal ethics standards.Sibutramine hyrdrochloride capsule was made in Fuling Pharmaceutical Factory of Taiji Group(batch number:X20010279).Electro-acupuncture apparatus(LH402A acupoint neutoelectrical stimulation equipment)was made in Beijing Huawei Chanye Developing Company.METHODS:This study was performed at the Laboratory of Acupuncture and Pharmacology,and Expefimental Animal Center,Nanjing University of Chinese Medicine from October 2004 to November 2006.①100 male SD rats underwent high-fat diet.Fifty-four alimentary obesity rats were randomly divided into electroacupuncture group,sibutramine group and obesity group.wim 18 rats in each group.Another 18 rats were regarded as a normal group.②Rats in the electroacupuncture group were given electrical stimulation(2/15 Hz,4 mA)at lateral Housanli and Neiting acupoints,contralateral alternation every other day.Rats were fixed with self-made binding instrument,and rats maintained a consciousness but not an intense struggle.The electricai stimulation lasted for 15 minutes every day,one day was an interval every 6 days.and the electroacupuncture was given for 49 days and 42 times in total.Rats in the sibutramine group were ingragastrically administrated with 0.2 g/L sibutramine hydrochloride(2.0 mg/kg/d).Rats in the control group and obesity groups were fixed the same as electroacupuncture group but not given electrical stimulation.MAIN OUTCoME MEASURES:①Obese marker:To detect body mass,body length,weight reduction rate and body mass/body fat mass,and to measure total cholesterol(TC),high density lipoprotein cholesterol(HDL-C)and triacylglycerol (TG)with biochemical chromatometry;②insulin sensitivity index:To measure fasting blood glucose,insulin and insulin sensitivity index with enzyme-linked immunosorbent assay;③adipocyte secreting regulatory substance level:To measure serum resistin,leptin,adiponectin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and its soluble receptor level with enzyme-linked immunosorbent assay.RESULTS:Among 120 rats,54 alimentary obesity rat models and 18 normal rats were included in the final analysis.①Obesemarker:Bodymass,bodyfatmass,cholesterol and TG levels in the obesity group were significantly higher than in the normal group(P＜0.01).Body mass,body fat mass,body mass/body fat mass,cholesterol and TG levels in the electroacupuncture group were significantly lower than in the obesity group(P＜0.01).TG level in the sibutramine group was significantly lower than in the obesity group(P＜0.01).Weight reduction rate in the electroacupuncture group was significantly higher than in the sibutramine group(P＜0.01).Body mass/body fat mass in the electroacupuncture group was significantly lower than in the sibutramine group(P＜0.01).②Insulin sensitivity:Blood glucose and serum insulin levels in the obesity group were significantly higher than in the normal group,and insulin sensitivity index was significantly lower than in the normal group(P＜0.01).Blood glucose and insulin levels in the electroacupuncture and sibutramine groups were significantly lower than in the obesity group,but insulin sensitivity index was significantly higher than in the obesity group(P＜0.01).③Adipocyte secreting regulatory substance level:Adiponectin in the electroacupuncture group was significantly higher than in the obesity group (P＜0.01).Resistin and TNF-αin the electroacupuncture group was significantly lower than in the obesity grollp(P＜0.01).Resistin level in the sibutramine group was significantly lower than in the obesity group(P＜0.01).CONCLUSION:The effect of electroacupuncture on abnormally secretary products of adipose tissues in obesity rats is superior to the effect of sibutramine.This may be one of important mechanisms of weight reduction by acupuncture and moxibustion,and prevention and cure of obesity and its related diseases.

Objective: To investigate the effect of Guangdong Shenqu (GSQ) on intestinal flora structure in mice with food stagnation through 16S rDNA sequencing. Methods: Mice were randomly assigned to control, model, GSQ low-dose (GSQL), GSQ medium-dose (GSQM), GSQ high-dose (GSQH), and lacidophilin tablets (LAB) groups, with each group containing 10 mice. A food stagnation and internal heat mouse model was established through intragastric administration of a mixture of beeswax and olive oil (1:15). The control group was administered normal saline, and the model group was administered beeswax and olive oil to maintain a state. The GSQL (2 g/kg), GSQM (4 g/kg), GSQH (8 g/kg), and LAB groups (0.625 g/kg) were administered corresponding drugs for 5 d. After administration, 16S rDNA sequencing was performed to assess gut microbiota in mouse fecal samples. Results: The model group exhibited significant intestinal flora changes. Following GSQ administration, the abundance and diversity index of the intestinal flora increased significantly, the number of bacterial species was regulated, and α and β diversity were improved. GSQ administration increased the abundance of probiotics, including Clostridia, Lachnospirales, and Lactobacillus, whereas the abundance of conditional pathogenic bacteria, such as Allobaculum, Erysipelotrichaceae, and Bacteroides decreased. Functional prediction analysis indicated that the pathogenesis of food stagnation and GSQ intervention were primarily associated with carbohydrate, lipid, and amino acid metabolism, among other metabolic pathways. Conclusion The digestive mechanism of GSQ may be attributed to its role in restoring diversity and abundance within the intestinal flora, thereby improving the composition and structure of the intestinal flora in mice and subsequently influencing the regulation of metabolic pathways.

OBJECTIVE: To investigate the effect of bactericidal/permeability-increasing protein(BPI) on the outcome of sepsis in mice and its possible mechanism. METHODS: Sepsis was induced by injection of 2x10(6) colony-formed unit E. coli J5 via the tail vein. BPI of 5 mg/kg or equal volume of normal saline(NS) were injected intravenously at the same time. Endotoxin and TNFalpha levels in serum were assayed using a chromogenic limulus amebocyte lysate test and ELISA respectively. RESULTS: Seventy-two hour survival rate of septic mice was significantly higher in the BPI group (15/18) than in the NS group(8/18, P<0.01). Serum endotoxin levels in the BPI group (1.3+/-0.3 and 0.7+/-0.4 &mgr;g/L) were significantly lower than those in the NS group (3.9+/-0.8 and 2.5+/-0.9 &mgr; g/L, P<0.01) 0.5 and 1 hour following injection of bacteria respectively. The peak levels of serum tumor necrosis factor-alpha(TNFalpha)in the BPI group (1.9+/-0.6 &mgr;g/L) were also markedly lower than those in the NS group (3.8+/-0.8 &mgr;g/L, P<0.01) 1.5 hours following bacterial injection. But there was no significant difference in blood bacterial count between the BPI and NS groups 0.5, 1.5 and 3.0 hours after injection of bacteria. CONCLUSIONS: BPI has a marked protective effect on E. coli sepsis, which might be related to its action against bacterial endotoxin and its inhibition of TNFalpha production in sepsis.

OBJECTIVE: To investigate the effect and mechanism of bactericidal/permeability-increasing protein (BPI) on sepsis induced by intra-abdominal infection in rats. METHODS: Cecal ligation and puncture (CLP) was made on 20 rats with sepsis induced by intra-abdominal infection. BPI or equal volume of physiological saline (PS) was intra-abdominally given immediately and 12 h after CLP, respectively (2.5 mg/kg of BPI each time). Plasma endotoxin levels were determined with limulus amebocyte chromogenic assay. RESULTS: (1) The survival time in BPI group was significantly higher than that in PS group. (2) The values of the mean arterial pressure (MAP), the left ventricular systolic pressure (LVSP), the isovolumic ventricular pressure (IP), and the maximal change of left intraventricular pressure (+/-dp/dtmax) in BPI group, although decreasing, were markedly higher than those in PS group. (3) Plasma glutamic-pyruvic transaminase (GPT) and urea nitrogen levels in BPI group, though increasing, were obviously lower than those in PS group. (4) There was no significant change of plasma endotoxin levels in BPI group, while plasma endotoxin levels markedly increased in PS group. CONCLUSIONS: BPI has obvious protective effect on sepsis induced by intra-abdominal infection, which might be related to its neutralization of endotoxin.

Objective:To investigate whether hepatitis B virus X protein (HBx) mediates the podocyte injury through reactive oxygen species (ROS) /nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) signaling pathway.Methods:HBx-overexpressing lentivirus was transfected into renal podocytes of mouse to mimic the pathogenesis of hepatitis B virus-associated glomerulonephritis. Podocytes were divided into the following five groups: blank control group (no special treatment), negative control group (transfected with control lentivirus), HBx overexpression group (transfected with HBx overexpression lentivirus), HBx overexpression+NLRP3 siRNA group (transfected with HBx overexpression lentivirus and NLRP3 siRNA), and HBx overexpression+ROS inhibitor group (transfected with HBx overexpression lentivirus and adding ROS inhibitor). The morphological changes of podocytes were observed with electron microscope. The generation of ROS was detected by dichlorodihydrofluorescein diacetate assay (DCFH-DA). Hoechst 33342 staining was used to observe the morphological and quantitative changes of podocyte nuclei. Enzyme-linked immunosorbent assay was used to detect caspase-1 activity, and the levels of lactate dehydrogenase, interleukin (IL)-1β and IL-18. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression levels of mRNA and protein of pyroptosis-related protein, such as NLRP3, apoptosis-associated speck-like protein containing card (ASC), caspase-1, IL-1β and IL-18. TUNEL staining and flow cytometer were used to detect the number of pyroptosis cells. Immunofluorescence staining was used to detect the expression levels of desmin and nephrin.Results:After successful infection of podocytes with HBx-overexpressing lentivirus, pyroptosis-related morphological changes in the cells were observed under electron microscope. The level of ROS in the HBx overexpression group was significantly higher compared to the negative control group ( P<0.05). Hoechst 33342 staining revealed condensed nuclei in the HBx overexpression group. TUNEL staining and flow cytometer demonstrated that podocytes underwent increased pyroptosis in the HBx overexpression group. The mRNA and protein expression levels of pyroptosis-related proteins such as NLRP3, ASC, caspase-1, IL-1β and IL-18 were up-regulated upon HBx overexpression (all P<0.05). Caspase-1 enzyme activity, lactate dehydrogenase and desmin expression levels were enhanced after HBx overexpression (all P<0.05). However, NLRP3 knockdown or addition of ROS inhibitors attenuated the pyroptosis and increased expression levels of pyroptosis-related proteins caused by HBx overexpression (all P<0.05). Conclusion:ROS/NLRP3 pathway plays an important role in HBx-induced podocyte pyroptosis.