OBJECTIVETo investigate the effect of Jiangtang Yishen Recipe (JTYSR) on high insulin induced cell proliferation of human glomerular mesangial cells (HMCs) and the expression of insulin receptor substrate 1 (IRS-1) and phosphatidylinositol-3-kinase (PI-3K).

METHODSHMCs were divided into 4 groups, i.e., the negative control group, the high insulin model group, the JTYSR group, and the LY294002 group. The concentration of insulin, JTYSR, and LY294002 was respectively confirmed by pre-experiment. Different culture solution was respectively added for different groups. RPMI1640 culture solution was added for HMCs in the negative control group, while HMCs in the rest 3 groups were cultured by 100 nmol/L insulin for 24 h. Meanwhile, HMCs from the JTYSR group and the LY294002 group were exposed to 125 mg/L JTYSR and 80 micromol/L LY294002 respectively for further 48 h. The proliferation of HMCs was detected by MTT and flow cytometry. The protein expression of IRS-1 and PI-3K in HMC was detected by immunohistochemical assay and Western blot. Results The proliferation of HMCs induced by high insulin could be significantly lowered, and the protein expression of IRS-1 and PI-3K could be down-regulated in the JTYSR group and the LY294002 group (P <0.01). Compared with the LY294002 group, the protein expression of IRS-1 and PI-3K could be slightly down-regulated in the JTYSR group (P <0.05).

CONCLUSIONJTYSR could lower high insulin induced proliferation of HMCs, and its mechanism might be related to insulin signaling pathway.

Cell Proliferation ; drug effects ; Chromones ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Insulin Receptor Substrate Proteins ; metabolism ; Mesangial Cells ; physiology ; Morpholines ; Phosphatidylinositol 3-Kinase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction

Objective To study the effect of different concentration of 1,25-dihydroxyvitamin D3[1,25(OH)2D3] on cell proliferation,differentiation and the expression of vitamin D receptor (VDR) in mouse MC3T3E1 osteoblast.Methods Osteoblast were cultured in medium with different concentrations of 1,25(OH)2D3.Incubated for 48 h,cell proliferation of osteoblast were examined by MTT reduction assay (mono-nuclear cell direc cytotoxicity assay),the osteocalcin (OC) levels in cell medium were detected by ELISA,and the expression of VDR mRNA and protein were examined by using SYBR Green real-time PCR and Western blot,respectively.Results 1.After incubation with 1,25(OH)2D3 for 48 h,the number of MC3T3E1 osteoblast was significantly less than that in control group(P0.05).3.SYBR Green real-time PCR and Western blot results showed that the expression of VDR mRNA as well as VDR protein of osteoblast in 10-8,10-9 mol/L experimental groups were significantly higher than those in control group (Pa0.05).Conclusions Cell proliferation of mouse osteoblast can be inhibited,while the cell differentiation was promoted by 1,25(OH)2D3.1,25(OH)2D3 up-regulated the expression of VDR in mouse osteoblast,which suggested that the VDR signal pathway may play some role in proliferation and differentiation of osteoblast.

Objective To investigate the apoptosis of T lymphocytes,the expression of Bcl-2, Fas on the peripheral blood T lymphocytes in end stage renal disease patients;and to explore the characteristics of Th1 /Th2 profile and the influence of dialysis membranes with different permeability on the apoptosis of T lymphocytes of maintenance hemodialysis patients.Methods The study included 10 non-dialyszed (ND)patients,45 maintenance hemodialysis patients with cellulose acetate (CA) membranes(13),low-flux polusulfone(PS-LF) membranes(16),high-flux polusulfone (PS-HF) membranes (16) and 8 healthy volunteers (C).The apoptosis of T lymphocytes,expression of Bcl-2,Fas on peripheral blood T lymphocytes cultured with phytohemagglutinin (PHA) stimulation for 24 hours were measured by flow cytometry and immunohistochemical.ELISA was performed for detecting the levels of IFN-?and IL-4 in culture supematants.Results In ESRD patients,the apoptosis of T lymphocytes was greater than that of group C.Group CA was greater than group PS-HF and group PS-LF (P＜0.05).The expression of Bcl-2 on T lymphocytes in ESRD patients was lower than that of group C (P＜0.05).There was negative correlation between the T lymphocytes apoptosis and Bcl-2. The expression of Fas on T lymphocytes in ESRD patients was greater than that of group C (P＜0.05), and it was positive correlated with T lymphocytes apoptosis.The level of IFN-?of ESRD patients was decreased significantly compared with that in group C (P＜0.05),and there was negative correlation between T lymphocytes apoptosis and IFN-?.IL-4 was increased in ESRD patients (P＜0.05) and it was positive correlated with T lymphocytes apoptosis.Conclusions The accelerated apoptosis of T lymphocytes in ESRD patients may be related to the expression of Bcl-2 and Fas of T lymphocytes.ESRD patients show a suppressed secretion of IFN-?and an increased secretion of IL-4. T lymphocytes apoptosis of maintenance hemodialysis patients is influenced not only by the biocompatibility but also by the permeability of the dialysis membrane.

OBJECTIVETo study the distribution of genotypes about alcohol dehydrogenase 2 (ADH2) and aldehyde dehydrogenase 2 (ALDH2) and its relationship with drinking-behaviors in Chinese Han healthy population as to providing a theoretic direction for filtering out high-risk and sensitive individuals and taking preventive measures to decrease the alcohol-related diseases.

METHODSUsing questionnaires to select subjects (201 persons, including men 104, women 97) for collecting blood samples and data about drinking-behaviors. Techniques of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the genotypes of ADH2 and ALDH2.

RESULTSHeterozygous ADH2 and homozygous ALDH2 were the two dominant ones (respectively 53.23%, 68.16%). There were no statistical differences among the distributions of nine combinations of ADH2 genotypes and ALDH2 ones. The difference of distribution of homozygous ALDH2 between males having high and middle drinking-frequencies seemed to be statistically meaningful.

CONCLUSIONThe proportion of individuals carrying about "susceptible genotypes of alcohol-related diseases" in Chinese Han healthy population should be more than one half (68.16%), which calls on reinforcing the surveillance and preventing the alcohol-related diseases. Correlation between genotypes of ADH2 and ALDH2 and alcohol-related diseases should be more important.

Adolescent ; Adult ; Aged ; Alcohol Drinking ; ethnology ; genetics ; physiopathology ; Aldehyde Dehydrogenase ; genetics ; metabolism ; Aldehyde Dehydrogenase, Mitochondrial ; Asian Continental Ancestry Group ; genetics ; China ; Drinking Behavior ; Ethanol ; metabolism ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Surveys and Questionnaires ; Young Adult

With human chromosomes,as antigen,anti—human—chromosome antibodies (AhChrA)were detected specifically from SLE patients sera by the methods of immunogold—silver staining(IGSS)and immnuofluorescenee tese (IFT)。To SLE,the sensitivity and specificity of serumAhChrA was 58.1%and98.5%respeetively in IGSS,34.9%and99.5%respectively in IFT。Boththe incidence and titer of AhChrA were found to be colsely related to the pathoactivity and thedamages of some important organs or tissues,such as kidney damage,abnormal immunity andhematocytopenia.A preliminary analysis of the antigen components reacting to AhChrA was alsoperformed。

Objective To observe apelin and its receptor (APJ) expressions in human osteoblasts and evaluate the effect of apelin on osteoblasts.Methods The expressions of apelin and APJ in human osteoblasts were tested by RT-PCR and Western blot.After human osteoblasts were treated with apelin,cell proliferation was measured by [~3H] thymidine incorporation and cell counting.Cell function was measured by alkaline phosphatase (ALP) activity,the secreted osteocalcin level and typeⅠcollagen production .The activation of signaling cascades was tested by Western blot.Small-interfering RNA (siRNA) to blockade APJ was applied to observe effects of apelin on cell proliferation and the activation of signaling cascades.Results Both apelin and APJ were expressed in human osteoblasts.Apelin increased the proliferation and did not show the influences on ALP activity, osteocalcin secretion and type I collagen production in human osteoblasts.Apelin induced activation of phosphatidylinositol-3 kinase (PI3K) downstream effector (Akt),but not mitogen-activated protein kinase (MAPK) such as c-jun N-terminal kinase (JNK),p38 and ERK1/2 in human osteoblasts.Suppression of APJ with siRNA or LY294002 (PI3K inhibitor) abolished the apelin-induced cell proliferation and the activation of Akt.Conclusion Human osteoblasts express apelin and APJ.Apelin stimulates the proliferation of human osteoblast via APJ/PI3K/Akt pathway,but has no effect on osteoblast differentiation.

In order to study the value of sequential and quantitative analysis of chimerism in determination of optional time of donor lymphocyte infusion (DLI) and prediction of efficacy of DLI, six patients with leukemias who relapsed or failed of engraftment were treated with DLI. Serial and quantitative analyses of donor chimerism (DC) both prior to and following DLI were performed by multiplex PCR amplification of STR markers (STR-PCR) and capillary electrophoresis with fluorescence detection. The results showed that at the time of relapse or graft rejection, STR-PCR indicated the decreasing donor chimerism in all six patients, at levels ranging from 27.3% to 85.7%. The declining value of DC (<90%) was detected in four patients at 26 days before relapse or graft rejection diagnosed clinically. Therefore the decrease of value of DC can be identified the high risk of relapse or graft failure and can be used to guide DLI implementation at early stage. In this study the clinical response were seen in two patients, the value of DC in these patients increased with convertion to a predominant donor profile (>90%) or converted to stable FDC shortly after DLI, while in the patients without clinical response, the level of DC decreased persistently or declined after transient increase. Three patients without response received second DLI. It is concluded that the monitoring of chimerism is proved to be a valuable to determine the optional time point of DLI and to early evaluate the efficacy of DLI. Furthermore, it can present a rational basis for treatment of intensification in the patients who did not respond to first-line DLI treatment.
Adolescent ; Adult ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphocyte Transfusion ; Recurrence ; Tissue Donors ; Transplantation Chimera

OBJECTIVETo investigate the level and significance of interleukin-8 (IL-8), soluble intercellular adhesion molecule (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) in patients peripheral blood (PB) during mobilization for peripheral blood stem cells harvesting.

METHODSThe levels of IL-8, sICAM-1 and sVCAM-1 in patients were dynamically assayed by ELISA during the mobilization procedure and the number of CD(34)(+) cell, white blood cell (WBC) and platelet (BPC) by flow cytometric analysis and hematometry respectively. Colony formation was assayed by using semisolid methycellulose culture.

RESULTSThere was a significant increase in plasma levels of IL-8 and both adhesion molecules [IL-8 (247.4 +/- 84.2) microg/L (P < 0.01); sICAM-1 (530.3 +/- 286.1) microg/L (P = 0.002 7); sVCAM-1 (575.3 +/- 350.4) microg/L (P = 0.001 3)] during the mobilization process; furthermore, IL-8 and sVCAM-1 concentration in the patient's plasma was paralleled to the numbers of CD(34)(+) cell, CFU-GM, WBC and BPC (P < 0.001).

CONCLUSIONThe levels of IL-8, sICAM-1 and sVCAM-1 in the patient's plasma were correlated to the PB number of CD(34)(+) cells, CFU-GM, WBC and BPC during the mobilization process. It suggested that analysis of IL-8, and sVCAM-1 dynamic changes may serve as markers for CD(34)(+) cells.

Adolescent ; Adult ; Enzyme-Linked Immunosorbent Assay ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged ; Treatment Outcome ; Vascular Cell Adhesion Molecule-1 ; blood

OBJECTIVETo investigate the effect of heme oxygenase/carbon monoxide (HO-1/CO) system on lipid deposition at aortic intima and the mechanism involved in hyperlipidemic rabbits.

METHODSTotally 32 rabbits, were divided into four groups. One group as control. Three groups for the following treatments: 1.5% cholesterol ration (Ch group, n = 8); 1.5% cholesterol ration plus HO-1 inducer hemin (Hm group, n = 8); and instead of hemin, the HO-1 inhibitor, zinc protoporphyrin IX (Zn group, n = 8) was given by injection into the abdominal cavity. Experiments were lasted for 12 weeks. Rabbit aortas were then isolated as the samples for histopathologic and ultrastructural examination. The protein expressions of HO-1 and endothelin-1 (ET-1) were investigated by immunohistochemical staining and Western blot analysis.

RESULTSComparing with the Ch group, rabbits of the Hm group showed a remarkably less extent of lipid deposition at the aortic intima [(17.9 ± 3.0)% vs (54.0 ± 4.2)%], and rabbits of the Zn group had a marked extent of lesion development [(61.1 ± 3.5)%]. Lipid deposition, endothelial damage and neo-intimal formation were less severe in rabbits of the Hm group than those in the Zn or Ch group, respectively. Comparing with the control group, rabbits of the Ch group showed a significant decrease of aortic NO production and cNOS activity. However, there were an enhancement of CO production and HO-1 activity (P < 0.01). Compared with Ch group, rabbits of the Hm group showed a remarkable elevation of aortic HO activity and CO production, whereas rabbits of the Zn group showed a marked decrease of both parameters. Compared with the Ch group, rabbits of the Hm group demonstrated a marked reduction of aorta ET-1 expression, whereas Zn group had a significantly higher ET-1 expression.

CONCLUSIONSModulation of HO-1/CO system may improve vascular endothelial function and inhibit smooth muscle cell proliferation in hypercholesterolemic rabbits, likely through a compensatory mechanism and a reduction of ET-1 expression, eventually leading to an inhibition of atherosclerotic plaque development.

Animals ; Aorta ; metabolism ; pathology ; Carbon Monoxide ; metabolism ; Cholesterol ; pharmacology ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; pharmacology ; Heme Oxygenase-1 ; antagonists & inhibitors ; metabolism ; Hemin ; pharmacology ; Hyperlipidemias ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Plaque, Atherosclerotic ; metabolism ; pathology ; prevention & control ; Protoporphyrins ; pharmacology ; Rabbits ; Tunica Intima ; metabolism ; pathology

BACKGROUNDHyperparathyroidism (HPT) occurs at an early age and has a high disability rate. Unfortunately, confirmed diagnosis in most patients is done at a very late stage, when the patients have shown typical symptoms and signs, and when treatment does not produce any desirable effect. It has become urgent to find a method that would detect early bone diseases in HPT to obtain time for the ideal treatment. This study evaluated the accuracy of high field magnetic resonance imaging (MRI) combined with spiral computed tomography (SCT) scan in detecting early bone diseases in HPT, through imaging techniques and histopathological examinations on an animal model of HPT.

METHODSEighty adult rabbits were randomly divided into two groups with forty in each. The control group was fed normal diet (Ca:P = 1:0.7); the experimental group was fed high phosphate diet (Ca:P = 1:7) for 3, 4, 5, or 6-month intervals to establish the animal model of HPT. The staging and imaging findings of the early bone diseases in HPT were determined by high field MRI and SCT scan at the 3rd, 4th, 5th and 6th month. Each rabbit was sacrificed after high field MRI and SCT scan, and the parathyroid and bones were removed for pathological examination to evaluate the accuracy of imaging diagnosis.

RESULTSParathyroid histopathological studies revealed hyperplasia, osteoporosis and early cortical bone resorption. The bone diseases in HPT displayed different levels of low signal intensity on T(1)WI and low to intermediate signal intensity on T(2)WI in bone of stage 0, I, II or III, but showed correspondingly absent, probable, osteoporotic and subperiosteal cortical resorption on SCT scan.

CONCLUSIONHigh field MRI combined with SCT scan not only detects early bone diseases in HPT, but also indicates staging, and might be a reliable method of studying early bone diseases in HPT.

Animals ; Bone Diseases ; diagnosis ; pathology ; Calcium ; blood ; Female ; Hyperparathyroidism ; complications ; Magnetic Resonance Imaging ; methods ; Male ; Osteoporosis ; diagnosis ; Phosphorus ; blood ; Rabbits ; Tomography, Spiral Computed ; methods