Objective To investigate the protective effects of angiotensin Ⅱ receptor antagonist losartan on diabetic nephropathy (DN).Methods 37 DN cases complicated by renal insufficiency were divided into two groups which were treated with losartan and enalapril,respectively.Blood pressure,plasma creatinine,blood urea nitrogen (BUN),uricemia,urinary protein and albumin were measured before and at 2,4,8,12 weeks after treatment.Results Losartan and enalapril lowered blood pressure remarkably (P0.05).Losartan decreased uricemia remarkably (P

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

BACKGROUND: The large surfaces of the skin are often initial site of contact between microorganism and human. The skin are coated with epidermis and epithelial cells can recognize microorganism and mount a fast defense through the production of various inducible antibiotic peptides. This leads to chracteristic broad spectrum of antimicrobial activity against bacteria, fungi, and viruses. Recent studies introduce us new peptides with antimicrobial activity such as P,-defensins and cathelicidins. They are expressed on the epithelia and polymorphonuclear leukocytes, which are first lines of defence from various invasive environments. Futhermore, they are considered very interesting and important endogenous antibiotics. Our previous study has shown that the expression of human defensin(hBD-2) mRNA, which is potent antibiotic peptide against Gram-negative bacteria(P. aeruginosa), was upregulated with ultraviolet(UV) irradiation, tumor necrosis factor-α(TNF-α) and lipopolysaccharide(LPS) in HaCaT cells. A novel hBD-3, 5-kDa, nonhemolytic antimicrobial peptide, was demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes in especially, multiresistant S. aureus. We have analyzed the expression patterns of hBD-3 in HaCaT cell lines. OBJECTIVE: This research have done in order to evaluate the expression and regulation of hBD-3 mRNA in human keratinocyte cell lines. METHODS: HaCaT cell lines were used to all culture experiments. Cultured human keratinocytes were stimulated with UV irradiation or TNF-α or LPS to determine whether hBD-3 mRNA production occurred. Reverse transcription-polymerase chain reaction (RT-PCR) was per-formed to amplify hBD-3 cDNA from stimulated keratinocytes in a time dependant manner, and densitometry was used to verify the specificity of RT-PCR amplication products. RESULTS: Expression of hBD-3 was upregulated with UV irradiation, TNF-α and LPS in Ha-CaT cells compared to control CONCLUSIONS: Human keratinocytes are capable to induce hBD-3 mRNA, as well as hBD-2, in response to UV irradiation, TNF-α and LPS. suggesting that these cells could play an important role against the bacterial infection and UV light damage in human skin.
Anti-Bacterial Agents ; Bacteria ; Bacterial Infections ; Cathelicidins ; Cell Line* ; Densitometry ; DNA, Complementary ; Epidermis ; Epithelial Cells ; Fungi ; Humans* ; Keratinocytes ; Necrosis ; Neutrophils ; Peptides ; RNA, Messenger ; Sensitivity and Specificity ; Skin ; Ultraviolet Rays

Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P

No abstract available.
Reye Syndrome*

AIMTo compare the TAOC of quercetin, rutin, vitamin C, vitamin E in vitro and examine the effect of quercetin on TAOC of rat plasma after intragastric administration.

METHODSFe3+ reducing ability assay, UV spectrum analysis and HPLC analysis were used to measure TAOC of plasma and the contents of quercetin and rutin after intragastric administration.

RESULTSThe TAOC of quercetin was stronger than that of rutin and roughly equal to vitamin C and vitamin E in vitro. After intragastric administration of quercetin (40 mg/kg bw), the TAOC and content of quercetin in rat plasma increased significantly. Vitamin C also increased plasma TAOC significantly, but rutin and vitamin E didn't after intragastric administration. However, there was no remarkable absorption peak of quercetin on HPLC chromatograms and on the other hand, the peak areas of two unknown peaks near quercetin peak were increased after intragastric administration of quercetin.

CONCLUSIONThe antioxidant capacity of quercetin was stronger than rutin and comparable to vitamin C both in vitro and in vivo. After absorption, quercetin is metabolized to its derivatives.

Animals ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Male ; Quercetin ; pharmacology ; Rats ; Rats, Wistar ; Rutin ; pharmacology ; Vitamin E ; pharmacology

AIM: To investigate the functional role of TGF-?_1 signal protein Smad2/3 in tubulointerstitial fibrosis associated with unilateral ureteral obstruction in rats. METHODS: The unilateral ureteral obstruction (UUO) model was induced by the ligation of left ureter. Rats were sacrificed at 1, 3, 7, 14, 21, and 28 days after UUO was initiated. TGF?_1 protein, phosphorylated Smad2/3 and interstitial ?-smooth muscle actin (?-SMA) expression were assayed by immunohistochemical staining. TGF-?_1 mRNA in the obstructed kidney was analyzed with in situ hybridration. HE and Masson staining were used for histological and morphometric studies of the pathological change in obstructed kidney. RESULTS: The results showed that upregulation of TGF-?_1 in tubulointerstitium of both cortex and medulla at day 3 (a 3.1 fold increase vs control, P

AIM:To investigate the expression and probable role of extracellular signal-regulated protein kinase(ERK1/2)in renal fibrosis associated with diabetic in mice.METHODS:Male homozygous C57BL/6 mice were divided at random into control group(intraperitoneally injected with citrate buffer)and diabetes group(received 5 consecutive daily intraperitoneal injections of streptozotocin at dose of 50 mg?kg-1?d-1).All mice were followed up for 16 weeks.Diabetes was confirmed by serum glucose levels exceeding 16.7 mmol/L.Mice were killed at 0,4,8,12 and 16 weeks respectively after streptozotocin injection.The kidney tissues were obtained from diabetic and control mice.Serum glucose,kidney weight/body weight(KW/BW),24 h albumin excretion rate(UAE)and the serum creatinine(Scr)were measured.The kidney tissue was used for histological and morphometric studies of glomerular size,glomerular matrix expansion(PAS),and the expression of TGF-?1,phosphorylated ERK1/2 and collagen Ⅲ by immunohistochemical staining.RESULTS:The serum level of glucose in streptozotocin-induced diabetic mice increased significantly.The kidney weight/body weight ratio,glomerular volume and glomerular matrix expansion in diabetic mice were obviously higher than those in control mice.Serum creatinine and 24 h albumin excretion rate in diabetic mice increased significantly compared with control mice.TGF-?1,phosphorylated ERK1/2 and collagen Ⅲ levels were obviously increased in the kidney of diabetic mice compared with those in control mice(P

Objective To prepare recombinant human epidermal growth factor (rhEGF) sustained-release microspheres and evaluate their morphology, rhEGF releasing activities and cell proliferation activity in vitro and compare difference of rhEGF sustained-release microspheres and rhEGF in facilitaring ulcer healing in diabetic rats. Methods (1) rhEGF sustained-release microspheres were prepared by the modified double emulsion method. Morphology of the microspheres was detected by transmission electron microscope and size distribution measured by laser granularity meter/Zeta electric potential meter. ELISA assays were applied to determine rhEGF releasing. (2)Proliferation of mouse fibroblasts was analyzed by MTr method. (3) Diabetic rat models were prepared and divided into four groups, ie, rhEGF sustained-release mierospheres group (Group A), rhEGF stock solution group (Group B), blank sustainedrelease mierospheres group (Group C) and PBS meustruum control group (Group D), which were given drug once a day. The wound healing rate was calculated by taking photographs at days 3,7,14 and 21. Skin specimens from the wound edge were harvested partially for observation of hydroxyproline (HYP) contents. Immunohistochemistry was employed to detect integrin 131 and keratin-19 and measure their positive staining area ratio. Results (1) The particle diameter of rhEGF sustained-release microspheres was 193.5 nm, with relative uniform particle diameter distribution. There showed no conglutination among rhEGF susrained-release microspheres, with good dispersibility. Releasing drug lasted for 24 hours and accorded with Higuchi release kinetic model. (2) Different concentrations of rhEGF sustained-release microspheres could promote the proliferation of mouse fibroblast, especially the concentration of 10 μg/L (P <0.05, compared with the control). (3) From the 7th day after treatment, Group A had the fastest wound healing rate, with statistical difference compared with other three groups (P < 0.05). Group A had higher HYP contents and positive area ratio of integrin β1 and keratin-19 than Group B. Conclusions rhEGF sustained-release microspheres prepared by the modified double emulsion method have uniform particle size and can last release for 24 hours. Compared with rhEGF stock solution, rhEGF sustained-release microspheres have faster and better ulcer healing and higher healing quality in diabetic rats.

Aged ; Coal Mining ; Electrocardiography ; Heart Rate ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; complications ; physiopathology ; Tachycardia ; etiology