Cell Differentiation ; drug effects ; Cell Line ; Connective Tissue Growth Factor ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; Lead ; toxicity ; Transforming Growth Factor beta ; metabolism

In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.

Tanreqing injection is suitable for early pneumonia, acute bronchitis, acute exacerbations of chronic, and upper respiratory tract infection which are classified with phlegm-heat obstructing lung syndrome of traditional Chinese medicine. To understand the clinical adaptation syndromes and medication characteristics of the post-market Tanreqing injection, the research team of the paper monitored the patients who are used with Tanreqing injection from September 2012 to October 2013 in four leader hospitals based on the method--prospective, multi-center, large sample, registration-type hospital centralized monitoring,and analyzes the general information, diagnostic information and medication characteristics of patients, in order to produce evidence for clinical practice and medication decisions and to establish the foundation of rational drug use.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; therapeutic use ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Prospective Studies ; Respiratory Tract Infections ; Young Adult

To determine the relationship between the urinary endothelin (ET-1), nitric oxide (NO) levels and the clinical, pathologic types of primary glomerulonephritis (GN) patients, urinary levels of ET-1 and NO were detected in 27 patients with biopsy-proven primary GN and 12 normal controls by radioimmunoassay and by copper-plated and cadmium column reduction assay, respectively. The results showed that urinary ET-1 levels in the patients with primary GN were significantly higher than in normal controls (p < 0.01), while the urinary ET-1 levels in patients with moderate mesangial proliferation GN were significantly higher than those in patients with mild mesangial proliferation GN (p < 0.05). Urinary ET-1 levels in patients whose clinical feature was nephrotic syndrome were found to be higher than in patients whose clinical feature was nephritic syndrome. However, urinary NO levels were to the contrary (p < 0.05). The ratio of ET-1/NO in primary GN patients was significantly higher than that in normal controls, and it positively correlated with the 24-hour urinary excretion of protein. These results suggest that urinary ET-1 levels are related to the proliferation of mesangial cells. The imbalance between ET-1 and NO may be related to the pathogenesis of primary GN and the occurrence of proteinuria.
Adolescence ; Adult ; Endothelin-1/urine* ; Endothelin-1/physiology ; Female ; Glomerulonephritis/urine* ; Glomerulonephritis/etiology ; Human ; Male ; Middle Age ; Nitric Oxide/urine* ; Nitric Oxide/physiology ; Nitric-Oxide Synthase/metabolism

ObjectiveToinvestigatetheinflammatoryresponseandapoptosisof peripheral blood mononuclear cells(PBMCs) and their regulation by triptolide(TP) in IgA nephropathy(IgAN) patients.MethodsBlood samples were collected from 29 IgAN patients and 16 healthy individuals.TNF-α and IL-6 concentrations were measured by ELISA and NO concentration by Griess reagent in the plasma of samples.PBMCs were isolated from IgAN patients and cultured in vitro,and subsequently activated by PHA(10 mg/L).The cytotoxicity of different TP concentrations was assayed by MTT and two non-toxic concentrations (12.5 μg/L or 25.0 μg/L)were selected for treatment.TNF-α,IL-6 and NO concentrations were measured in the culture media collected from PBMCs cultures activated by PHA (10 mg/L) and treated with TP (12.5 μg/L or 25.0 μg/L).The PHA-activated,TP-treated cells apoptotic rate was analyzed by FACS using Annexin V-FITC staining.The expression of Bcl-2,Bax,caspase-9 and caspase-3 were detected by RT-PCR and Western blotting from lyses of PHA-activated with or without TP-treated cells.ResultsThe serum concentrations of TNF-α[(131.57±50.61) ng/L vs(30.24±18.93) ng/L,P＜0.01],IL-6[(76.36±25.21) ng/L vs(35.08±16.59) ng/L,P＜0.01] and NO[(46.36±12.93) μmol/Lvs (26.61 ±10.87) μmol/L,P＜0.01] were significantly increased in IgAN patients compared to healthy individuals.PBMCs viability in culture decreased after TP treatment in a dose-dependent manner.TP also inhibited TNF-α,IL-6 and NO levels in the media of PHA-activated PBMCs in culture and induced PBMCs apoptosis.The expression of Bcl-2 decreased markedly and Bax,caspase-9 and caspase-3 increased significantly after TP treatment (all P＜0.05).Conclusions The PBMCs from IgAN patients are in a highly activated state,and have a high apoptotic rate.TP treatment induces benificial effects in IgAN patients by inhibiting the activation of PBMCs by activating pro-apoptotic pathway.

OBJECTIVE: To determine the effect of indomethacin on high concentration glucose and lipopolysaccharide (LPS) induced fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) secretion in cultured human peritoneal mesothelial cells. METHODS: Mesothelial cells were isolated from human omental specimens by trypsin disaggregation and incubated by 2.5% glucose or LPS together with different concentrations of indomethacin. Enzyme-linked immunosorbent assay determined the quantity of FN and PAI-1 in the cultured supernatants. RESULTS: Compared with the control group, the levels of FN and PAI-1 in the cultured supernatants were increased significantly exposuring to high concentration glucose and LPS (P <0.01). The different concentrations of indomethacin decreased FN and PAI-1 secretion in the 2.5% glucose or the LPS group within 24 h (P < 0.05). CONCLUSION Indomethacin can inhibit the synthesis and secretion of extracellular matrix in human peritoneal mesothelial cells, which may be effective in the gene therapy for peritoneal fibrosis.
Cells, Cultured ; Cyclooxygenase Inhibitors ; pharmacology ; Epithelial Cells ; metabolism ; Fibronectins ; metabolism ; Humans ; Indomethacin ; pharmacology ; Peritoneum ; cytology ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism

OBJECTIVE: To analyze the etiology and the relevant factors such as age, sex, blood pressure, outcomes and causes of death in end stage renal disease (ESRD) with hemodialysis (HD) in some hospitals in Hunan province. METHODS: The retrospective analysis included 1,622 ESRD with HD patients. Data on the etiology, demographic and epidemiologic aspects of these patients were examined, and life expectancy and mortality rate were calculated. RESULTS: In 1,622 ESRD with HD patients, the average age at the start of HD was 46.91 +/- 15.41, and the male/female ratio was 1.45/1. As the leading cause, chronic glomerulonephritis accounted for 56.43%, followed by hypertensive nephropathy (12.58%), obstructive nephropathy (9.13%) and diabetic nephropathy (8.85%). In recent years, the constituent ratio of diabetic nephropathy rose. The number of ESRD maintenance HD (MHD) patients was 581. Among them, 43.7% remained MHD, 13.0% received renal transplantation, 19.9% were transferred to other hospitals for HD, 7.2% became peritoneal dialysis, 14.8% died, and 1.4% ceased treament for economic reasons. The longest MHD was 13 years. The 1st-year, 3rd-year and 5th-year survial rate of MHD patients was 93.53%, 68.92% and 62.51%, respectively. The leading cause of death was cardiovascular incidence. In this group of ESRD with (53.6%), and then cerebrovascular disorder (21.0%). CONCLUSION HD patients, the age of starting dialysis was 30 approximately 70. The first cause was chronic glomerulonephritis. As the age increased, the constituent ratio of diabetic nephropathy rose. In MHD patients, the 1st-year, 3rd-year and 5th-year survial rate of maintenance hemodialysis patients was 93.53%, 68.92% and 62.51%, respectively. The first cause of death was cardiovascular accidence, and then cerebrovascular disorder.
Adult ; Aged ; China ; Diabetic Nephropathies ; complications ; therapy ; Female ; Glomerulonephritis ; complications ; therapy ; Humans ; Hypertension, Renal ; complications ; Kidney Failure, Chronic ; etiology ; therapy ; Male ; Middle Aged ; Renal Dialysis ; Retrospective Studies

OBJECTIVE: To investigate the effect of the peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonist troglitazone on TGF-beta(1) and fibronectin (Fn) expression in human peritoneal mesothelial cells (HPMCs). METHODS: HPMCs were cultured from human omentum by an enzyme digestion method, growing in medium containing 30 mmol/L D-glucose. TGF-beta(1) and Fn expression were measured in HPMCs in the presence and absence of 15 micromol/L troglitazone. The mRNA expressions of PPAR-gamma,TGF-beta(1) and Fn were determined by semi-quantification reverse transcriptive PCR (RT-PCR). The protein of TGF-beta(1) was determined by enzyme-linked immunosorbent assay (ELISA) and proteins of PPAR-gamma and Fn were determined by Western blot. RESULTS: The mRNA and protein expression of TGF-beta(1) and Fn were significantly increased in HPMCs stimulated with 30 mmol/L D-glucose compared with the control group with F12 media (P<0.01). Obvious decrease of TGF-beta(1) was found in troglitazone(15 micromol/L) treated group compared with group stimulated with 30 mmol/L D-glucose (P<0.05). Exposure of HPMCs to troglitazone reduced the Fn secretion (P<0.05). CONCLUSION Troglitazone reduced the expression of TGF-beta(1) in HPMCs stimulated by 30mmol/L D-glucose, and reduced Fn production. PPAR-gamma agonists may have a specific role in ameliorating the course of progressive peritoneal fibrosis under long-term peritoneal dialysis states.
Blotting, Western ; Cells, Cultured ; Chromans ; pharmacology ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Fibronectins ; biosynthesis ; genetics ; Glucose ; pharmacology ; Humans ; PPAR gamma ; biosynthesis ; genetics ; Peritoneum ; cytology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta1 ; biosynthesis ; genetics ; Troglitazone

OBJECTIVE: To observe the changes of vasoactive substances in rabbits administered with mannitol at different dosages and to investigate the mechanism of acute renal failure (ARF) induced by massive mannitol administration. METHODS: Eighteen healthy male New Zealand rabbits were randomly divided into 3 groups: a minor mannitol group (n=6, mannitol 8 g/kg within 2 hours), a control group (n=6, saline of the same volume), and a massive mannitol group with free water taking (n=6, mannitol 40~60 g/kg within 3 days). The changes of renin, angiotensin-I (ang-I), angiotensin-II (ang-II), endothelin (ET), and atrial natriuretic factor(ANF) in the serum were observed. RESULTS: No significant changes in the renin, ang-I, ang-II, ET, and ANF in the serum were found between the minor mannitol group and the saline control group (P> 0.05). In the massive mannitol group with free water taking, renin, ang-I, and ang-II in the serum increased significantly compared with the other 2 groups; ET in the serum decreased significantly compared with the saline control group (P< 0.05); no significant changes in the ANF in the serum were found compared with the other 2 groups(P> 0.05). CONCLUSION ARF induced by massive mannitol administration is associated with a significant change of vasoactive substances.
Acute Kidney Injury ; blood ; chemically induced ; physiopathology ; Angiotensins ; blood ; Animals ; Atrial Natriuretic Factor ; blood ; Dose-Response Relationship, Drug ; Endothelins ; blood ; Male ; Mannitol ; administration & dosage ; toxicity ; Rabbits ; Random Allocation ; Renal Circulation ; drug effects ; Renin-Angiotensin System ; drug effects

OBJECTIVE: To construct the expressing vector of siRNA which can inhibit the Smad3 activity. METHODS: Sixty-four bases of 2 pair oligos for hairp in RNA expression which targeted Smad3 gene were chemically synthesized and annealed. pSUPER vector was linearized with BgL II and Hin d III treated with alkaline phosphatase (CIP). Anneled oligos were inserted into the downstream of the treated pSUPER's pol III H1 promoter to construct RNAi plasmid (pSUPER Smad3). Oligos with a scrambled sequence were used as a negative control. pSUPER Smad3 was transfected into human renal tubular epithelial cells (HKC). RESULTS: Recombinant pSUPER Smad3 vector was identified by the digestion with Eco R I and Hin d III, and confirmed by the sequencing analysis with T3 primer. Sixty-four bases had been inserted into the expected site. Furthermore, the insertion sequence was exactly corrected. The activity evaluation indicated that mRNA and protein of Smad3 but not Smad2 were inhibited by pSUPER Smad3 in HKC. CONCLUSION The pSUPER Smad3 system has been constructed successfully, and has high inhibition and specificity in vitro.
Epithelial Cells ; metabolism ; Humans ; Kidney Tubules ; cytology ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Smad3 Protein ; antagonists & inhibitors ; genetics ; Transfection