OBJECTIVETo observe the renal protection effects of Compound Shenhua Tablet (CST) on diabetic nephropathy (DN) rats.

METHODSDN rats were given a normal diet for 9 months after they were induced by intraperitoneal injection of STZ at the dose of 65 mg/kg after uninephrectomized. They were randomly divided into 4 groups, i. e., the normal control group, the model control group, the CST group, and the Irbesartan group. The intervention was given by gastrogavage for 6 weeks. The general state, 24 h urine protein, urine micro-albumin (mAlb), serum creatinine (SCr), blood urea nitrogen (BUN), glucose (GLU), triglyceride (TG), total cholesterol (TC), total protein (TP), and albumin (ALB) levels were observed before and after intervention. Renal pathological changes were observed by PAS staining and transmission electron microscope.

RESULTSAfter 6 weeks of drug intervention, when compared with the model control group, the general state was improved in the CST group and the Irbesartan group. The levels of 24 h urine protein, urine mAlb, SCr, BUN, GLU, TG, and TC were obviously lower in the CST group and the Irbesartan group than in the model group as well as in the same group before treatment (P<0.05, P<0.01). There was no statistical difference between the two treatment groups (P>0.05). The renal pathological changes and the renal ultrastructure were improved to some degree in the two groups when compared with those in the model control group.

CONCLUSIONSCST could attenuate the renal damage of diabetes and delay renal deterioration process. Its effectiveness was equivalent to that of Irbesartan.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the activation and proliferation of specific T cells induced by artificial antigen-presenting cells (aAPCs) simulated dendritic cells (DCs) and to observed the effect of these T cells on leukemic cell killing.

METHODSaAPCs were developed by coating a human leukocyte antigen-immunoglobulin fusion protein ( HLA-lg), which was connected each one of the four CML28 antigen epitopes (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV), and CD28-specific antibody, to magnet-beads CML cell specific peptides (CML28) served as target peptides. Bone marrow (BM) or peripheral blood (PB) mononuclear cells (MNCs) were isolated from HLA-A2 healthy volunteers, and co-cultured with aAPCs. Specific T lymphocyte were detected by flow cytometry. The fresh acute leukemic cells were used as target cells. The specific T cells incubated with leukemic cells for 4 h at ratios of 5:1, 10:1, 20:1, 40:1, 80: 1, respectively. The effect of leukemic cells killing was detected by lactate dehydrogenase release test.

RESULTSThe average ratio of CML-28 specific T lymphocyte in control group was (2.2 +/- 0.4)% and in experimental groups (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV) were (13.5 +/- 1.6)%, (15.2 +/- 1.5)%, (14.7 +/- 1.8)% and (34.3 +/- 3.5)%, respectively, being significantly higher than that in control group (P < 0.01). Induction efficiencies of acute leukemic cells killing were significantly enhanced by increase of effector cells. The cytotoxic activity of specific T lymphocyte in one experimental group (VLTFALDSV) was much higher than that in other three experimental group (P < 0.05).

CONCLUSIONThis "prime and expand" regimen should be an alternative method for large scale amplification of rare tumor-specific CTLs and aAPCs might be a useful tool for leukemia immunotherapy.

Adolescent ; Adult ; Antigen-Presenting Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Female ; Humans ; Leukemia ; immunology ; pathology ; Lymphocyte Activation ; Male ; Middle Aged ; T-Lymphocytes ; cytology ; immunology ; Young Adult

This study was aimed to investigate the effect of artificial antigen-presenting cells (aAPCs) on inducing activation and proliferation of specific T-lymphocytes through stimulating biological function of dendritic cells with aAPCs in vitro. The specific antigen of chronic myeloid leukemia CML-28 was screened as objective antigen peptide by using magnetic microbeads as vector; the CML-28 epitope sequence (Vltfaldsv) was obtained by antigen epitope prediction software; this epitope was coupled with MHC molecule and used as first signal molecule, the B7-1 molecule was used as second signal molecule; these 2 molecules simultaneously were loaded onto surface of magnetic microbeads so as to contract aAPC complex. The bone marrow mononuclear cells were derived from HLA-A2(+) healthy bone marrow donors, CD8(+) T lymphocytes were screened and co-cultured with aAPC complex. During culture the 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) was added and proliferation of T-lymphocytes was detected by CPSE and proliferation level of specific T lymphocytes was detected by flow cytometry. The results showed that the proliferation level of CML-28 specific T lymphocytes obviously increased in experimental group, average level was 17.34 +/- 0.65%, while average level in control was 2.25 +/- 0.43%, there was significant difference between them (p < 0.01). It is concluded that the aAPC complex can mimic human APCs in vitro, and stimulate activation and proliferation of CD8(+) specific T lymphocytes.
Adolescent ; Adult ; Antigen Presentation ; immunology ; Antigen-Presenting Cells ; immunology ; metabolism ; Antigens, Neoplasm ; immunology ; Biomimetics ; methods ; CD8-Positive T-Lymphocytes ; immunology ; Female ; HLA-A2 Antigen ; immunology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; Lymphocyte Activation ; Male ; Middle Aged ; Tissue Donors ; Young Adult

OBJECTIVETo investigate the efficacy and safety of diisopropylamine dichloroacetate in the treatment of nonalcoholic fatty liver diseases (NAFLD).

METHODSA randomized, double-blind, dose-paralleled control trial was carried out with NAFLD patients. The patients were randomly assigned to 2 groups treated with either a high dosage (120 mg/d) or a low dosage (60 mg/d) of diisopropylamine dichloroacetate for 8 weeks and the efficacy and safety of the drug were examined.

RESULTS127 cases were recruited for the trial, 63 in the high dosage group, and 64 in the low dosage group. No case dropped out in the trial but four cases were eliminated (4/127, 3.1%). The final number in this trial was 123, with 61 in the high dosage group and 62 in the low dosage group. After 8 weeks of treatment, the overall improvement of clinical symptoms in the high dosage and in the low dosage group was 87.8% and 79.6%, respectively. ALT normalization was found in 55.7% and 69.4% of the cases in the two groups, serum lipids were lowered in 67.2% and 67.7% and ultrasound grading of the liver alteration severity was lowered in 51.7% and 43.5% in the two groups. The differences found between the two groups were of no statistical significance. One case from each group was found having an adverse drug reaction of dryness of the mouth (1.6%). No severe adverse drug reactions were found.

CONCLUSIONDiisopropylamine dichloroacetate could be used as a safe and effective drug in the treatment of nonalcoholic fatty liver diseases.

Adult ; Double-Blind Method ; Fatty Liver ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Quaternary Ammonium Compounds ; administration & dosage ; adverse effects ; therapeutic use

OBJECTIVETo determine differences in adherence to secondary prevention guidelines (pharmacological interventions) among coronary heart disease (CHD) patients between a Chinese medicine (CM) hospital and a general hospital in a Chinese city.

METHODSMedical records of 200 patients consecutively discharged from the CM hospital and the general hospital for CHD were reviewed to determine the proportions of eligible patients who received antiplatelet agents, β-blockers, angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs) and statins at discharge. The effects of patient characteristics and hospital type on the use of these medicines were estimated using logistic regression models.

RESULTSPatients discharged from the CM hospitals were older; more likely females; had greater history of hyperlipidemia, cerebrovascular diseases and less smoker (P<0.01 or P<0.05). They were less likely to receive coronary angiography and percutaneous coronary intervention, and had a longer length of stay than those discharged from the general hospital (P<0.01 or P<0.05). There were no significant differences in antiplatelet agents (96% vs. 100%, P=0.121) or statins (97.9% vs. 100%, P=0.149) use between the CM hospital and the general hospital. In multivariable analyses that adjusted for patient characteristics and hospital type, there was no significant difference in use of β-blockers between the CM hospital and the general hospital. In contrast, patients discharged from the CM hospital were less likely to receive ACE inhibitors/ARBs compared with those discharged from the general hospital (odds ratio: 0.3, 95% confidence interval: 0.105-0.854).

CONCLUSIONIn this study, the CM hospital provides the same quality of care in CHD for prescribing evidence-based medications at discharge compared with another general hospital except for ACE inhibitors/ARBs use.

Aged ; Coronary Disease ; drug therapy ; prevention & control ; Evidence-Based Medicine ; Female ; Hospitals, General ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Secondary Prevention

Objective To detect the hepatotoxicity biomarkers using normal human hepatocyte (HepaRG) and high-content screening,and to combine the micronucleus test and single cell gel electrophoresis to estalish a rapid screening platform for in vitro cytotoxitity and genotoxicity.Methods The effects of rhubarb anthraquinones (AQs) on the reactive oxygen species (ROS),intracellular Ca2+ concentration and mitochondrial membrane potential (MMP) in HepaRG cells were studied using appropriate fluorescent probes Hoechst33342、DCFH-DA、Fluo4-AM、Mito Tracker Red CMX Ros and high-content screening methods,and the potential genotoxiciy triggered by AQs were analyzed using the high-content based cytokinesis block micronucleus test and high throughput comet assay.Results The intracellular ROS level of HepaRG cells was significantly elevated by a 24 h treatment with Emodin (25.0 μg/mL),aloe-emodin (25.0 μg/mL) or chrysophanol (50.0 μg/mL),which are dose-concentration dependent (P ＜ 0.05 and 0.01);the intracellular Ca2+ increased and mitochondrial damage were observed with the treatment of aloe-emodin (25.0 μg/mL) and rhein (50.0 μg/mL,P ＜ 0.05 and 0.01).Comparing to control group,Emodin (25.0 μg/mL) induced an increased micronucleus rate (1.59％ ± 0.68 ％,P ＜ 0.01) and significantly higher percentage tail DNA and Olive tail moment (respectively 10.155％ ± 2.17％ and 0.510 ± 0.06,P ＜ 0.05 and 0.01) after 24 h;while the chrysophanol increased the micronucleus rate to 1.29％ ± 0.54％ (P ＜ 0.01) after 72 h.Conelnsion The results on the cytotoxicities and genotoxicities of AQs are consistent with the literatures.In this study,a rapid screening model for both hepatotoxicity and genotoxicity was successfully established,which will help with the early screening during the drug development stage.

Objective To establish the systemic lupus erythematosus (SLE) mouse model through pristaneip injection and validate the model comprehensively.Methods Female BALB/c mice of 6-8 weeks were randomly divided into two groups.Animals in model group were injected with 0.5 mL pristane by ip injection while in control group with 0.5 mL normal saline.Anti-systemic lupus erythematosus antibodies (anti-SLE) and anti-double strand DNA antibodies (anti-dsDNA) were checked before injection and monthly thereafter.Proteinuria was detected before injection and every month thereafter.All mice were bled to death 6 months after injection.Kidneys were excised to observe the histopathologic evidence of glomerulonephritis.Results The concentration of anti-dsDNA and anti-SLE antibody in sera was higher of model group than that of control group two months after pristane injection,and the concentration of antibody gradually increased within 6 months.At the sixth months,the protein concentration of urine in most model group mice was ++++.The histopathology and imunoflorescence of kidney sections indicated typical evidence of glomerulonephritis in model group.Conclusion The murine lupus model can be successfully established in female BALB/e mouse with a single ip injection of 0.5mL ofpristane.

OBJECTIVEGlucocorticoid (GC) is the first therapeutic choice of primary nephrotic syndrome (PNS). The response to GC treatment is an important indicator for the outcome of PNS children. Children with GC-resistant PNS present with incomplete or no response to GC, and may herald the progression to end-stage renal failure. However, the detailed mechanism of GC-resistance or GC-sensitive effect in these PNS children has not been clearly elucidated. The previous study by the authors indicated that there was increased expression of GR beta in PBMCs in GC-resistant children with PNS, and the over expression of GR beta resulted in GC resistance via influencing the ability of GR alpha nuclear translocation. To elucidate the relationship between GR beta expression in renal and in PBMCs and the effect of glucocorticoid on glucocorticoid-resistance children with PNS, the expression of GR alpha and GR beta in renal tissue and in PBMCs were detected by immunohistochemistry.

METHODSForty children with PNS were divided into two groups, GC-resistant group(20) and GC-sensitive group(20), the expression of GR alpha and GR beta in renal intrinsic cells and in PBMCs were measured with the immunohistochemistry technique. A semiquantitative score was used to evaluate the injury degree of the glomeruli and tubulointerstitium.

RESULTSCompared with GC-sensitive group, the glomerular pathologic scores (6.91 +/- 1.98) and renal tubular pathologic scores (7.12 +/- 1.62) in GC- resistant group were significantly different (P < 0.01, respectively). GR alpha expressions of renal tissue and PBMCs were higher in the control group (58.3 +/- 2.6, 59.1 +/- 7.2) than those in the GC-sensitive group (40.2 +/- 7.2 and 36.6 +/- 5.1, P < 0.01, respectively) and GC-resistant group (35.0 +/- 8.2 and 36.4 +/- 6.6, P < 0.01, respectively). GR beta expressions of renal tissue and PBMCs were higher in the GC-resistant group (13.8 +/- 3.0 and 12.1 +/- 4.1) and in the GC-sensitive group (6.5 +/- 1.9 and 5.9 +/- 1.0) than that in control group (2.3 +/- 0.4 and 3.2 +/- 1.1, P < 0.01, respectively). GR beta expressions in renal tissue and PBMCs were higher in the GC-resistant group than that in the GC-sensitive group (P < 0.01). Compared with control group, GR beta expressions in PBMCs and in renal tissue were lower than those in mild renal lesion group (5.4 +/- 2.8, 6.46 +/- 2.50), midmedium renal lesion group (8.7 +/- 2.4 and 11.4 +/- 3.7) and (17.1 +/- 0.4 and 18.7 +/- 0.7) in severe renal lesion group (F = 5.8, 15.6, P < 0.01, respectively). GR beta expression of PBMCs had a positive correlation with GR beta expression of renal intrinsic cells (r = 0.651, P < 0.01). GR beta expressions by PBMCs and renal intrinsic cells were positively correlated with renal pathologic scores (r = 0.579 and 0.623, P < 0.01, respectively).

CONCLUSIONGC-resistant children with PNS were related to the increased GR beta expression in PBMCs and renal intrinsic cells. There was no correlation between the GR alpha expressions in PBMCs and in renal intrinsic cells. Increased GR beta expression might decrease the effect of GC via inhibiting the activity of GR alpha.

Adolescent ; Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Kidney Glomerulus ; pathology ; Kidney Tubules ; pathology ; Male ; Nephrotic Syndrome ; drug therapy ; pathology ; Receptors, Glucocorticoid ; analysis

BACKGROUNDBivalirudin was widely used as an anticoagulant during coronary interventional procedure in western countries. However, it was not available in China before this clinical trial was designed. This randomized, single-blind and multicenter clinical trial aimed to evaluate the efficacy and the safety of domestic bivalirudin during percutaneous coronary intervention (PCI).

METHODSA randomized, single-blind, multicenter trial was designed. Elective PCI candidates in five centers were randomized into a bivalirudin group and a heparin group, which were treated with domestic bivalirudin and non-fractional heparin during the PCI procedure. The efficacy was evaluated by comparing the activated coagulation time (ACT), the procedural success rate (residual stenosis < 20% in target lesions without any coronary artery related adverse events within 24 hours after PCI), and the survival rate without major adverse cardiac events at 30 days after PCI between the two groups. Safety was evaluated by the major/minor bleeding rate.

RESULTSA total of 218 elective PCI patients were randomized into a bivalirudin group (n = 110) and heparin group (n = 108). Except for two patients needing additional dosing in the heparin group, the ACT values of all other patients in both groups were longer than 225 seconds at 5 minutes after the first intravenous bolus. Procedural success rates were respectively 100.0% and 98.2% in the bivalirudin group and heparin group (P > 0.05). Survival rates without major adverse cardiac events at 30 days after PCI were 100.0% in the bivalirudin group and 98.2% in the heparin group (P > 0.05). Mild bleeding rates were 0.9% and 6.9% (P < 0.05) at 24 hours, and 1.9% and 8.8% (P < 0.05) at 30 days after PCI in the bivalirudin group and heparin group respectively. There was one severe gastrointestinal bleeding case in the heparin group.

CONCLUSIONSDomestic bivalirudin is an effective and safe anticoagulant during elective PCI procedures. The efficacy is not inferior to heparin, but the safety is superior to heparin.

Aged ; Antithrombins ; adverse effects ; therapeutic use ; Female ; Heparin ; therapeutic use ; Hirudins ; adverse effects ; Humans ; Male ; Middle Aged ; Peptide Fragments ; adverse effects ; therapeutic use ; Percutaneous Coronary Intervention ; Recombinant Proteins ; adverse effects ; therapeutic use ; Single-Blind Method ; Survival Rate ; Whole Blood Coagulation Time

Protein polypeptides are a class of bioactive substances that play a crucial role in maintaining the stability of various functions of the organism.Rapid and accurate detection of their levels could help in the diagnosis of diseases,the monitoring of drug therapy and the research and development of medicines,which is of great significance in the fields of clinical medicine,biology and pharmacy.Conventional protein polypeptides detection methods,such as Western blotting and enzyme-linked immunosorbent assay,still have problems like low sensitivity or difficulty in determining more than 1 analyte simultaneously.Liquid chromatography-mass spectrometry(LC-MS)has the advantages of specificity,sensitivity and high throughput.However,low ionization efficiency of macromolecules and strong biological matrix effects limit the feasibility of direct detection of protein polypeptides by LC-MS,which has led to the development of signal conversion and amplification strategies based on LC-MS technology.In this review,we summarized the research progress of sample pretreatment methods and signal conversion and amplification strategies for quantification of protein polypeptides in vivo based on LC-MS in recent years,providing a reference for developing specific high-sensitivity detection methods for low-abundance protein polypeptides in complex samples based on LC-MS technology.