AIM: To investigate the effect of epidermal growth factor receptor (EGFR) inhibitor erlotinib on kidney injury in diabetic nephropathy (DN) rat and the underlying mechanism.METHODS: The rat model of DN was induced by intraperitoneal injection of streptozotocin (STZ) at dose of 55 mg/kg.One week after STZ injection, the rats with blood glucose level exceeding 16.7 mmol/L were identified as diabetic.Diabetic rats were randomly divided into 2 groups: STZ group and STZ+erlotinib group.In addition, the normal rats were used as control group.The rats in STZ+erlotinib group were treated with erlotinib at 100 mg·kg-1·d-1 for 4 weeks(5th~8th week).The fasting blood glucose (FBG), serum creatinine (SCr) and 24 h urine protein were measured.The pathological changes of the kidney were observed by HE staining and Masson staining.The protein levels of EGFR, p-EGFR, transforming growth factor β1 (TGFβ1), Smad2/3, p-Smad2/3, collagen Ⅳ (ColⅣ) and fibronectin in the kidney tissues were determined by Western blot.The reactive oxygen species (ROS) level and malondialdehyde (MDA) content in the renal tissues were futher analyzed.RESULTS: Compared with control group, the levels of FBG, 24 h urine protein and Scr were significantly increased in STZ group (P<0.01).Compared with STZ group, the levels of FBG, 24 h urine protein and SCr in STZ+erlotinib group were markedly decreased (P<0.05).In additon, the glomerular structure was restored to normal, the proliferative degree of mesangial cells markedly attenuated, and the epithelial cells were in alignment in STZ+erlotinib group.Moreover, erlotinib significantly inhibited the protein levels of p-EGFR, TGFβ1, p-Smad2/3, ColⅣ and fibronectin in the kidney tissues of STZ rats.In addition, erlotinib also significantly inhibited the levels of ROS and MDA in the kidney tissues of STZ rats.CONCLUSION: Erlotinib ameliorates STZ-induced diabetic nephropathy possibly through inhibiting the activation of EGFR/TGFβ1-Smad2/3 signaling pathway in association with suppression of fibrosis and oxidative stress.

Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma. Methods: Balb/c mice were immunized i. p . with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes ( VH and VL) of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA. , the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3. 5 ? 106 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified , which paves a way for study of prostate carcinoma.

This study is to screen the Chinese herbal compounds which could inhibit the production of Abeta and investigate the underlying mechanism. Ten types of compounds which have potential value in the treatment of AD were selected as initial screening trial. The cell models which used could overexpress Abeta and beta-secretases or Abeta and gamma-secretases. Extracellular Abeta was determined by ELISA after the cell models treated with different concentrations of compounds (0.5-100 micromol x L(-1)), separately. Then the compounds were selected which could inhibit extracellular Abeta and their best concentration ranges were decided, too. Furthermore, the cell viability and apoptosis rate, the level of intracellular Abeta, beta and gamma-secretases were determined after the cell models treated with different concentrations of selected compounds. The results showed that 4 of the 10 compounds could reduce the level of extracellular Abeta; they were cryptotanshinone, astragalosides, gastrodin and paeoniflorin, and their best concentration ranges were 0.5-5.0, 0.5-5.0, 5.0-50, 1.0-25 micromol x L(-1), respectively. Further study indicated that the 4 selected compounds were nontoxic to the cellular models and lowering intracellular Abeta were more effective compared with extracellular; of which astragalosides and gastrodin showed dose-dependent inhibition to the activities of beta and gamma-secretases, with the maximum inhibiting rates of 78.2% and 80.3%, respectively. In conclusion, cryptotanshinone, astragalosides, gastrodin and paeoniflorin could inhibit the expression and secretion of Abeta, and the underlying inhibiting mechanism of astragalosides and gastrodin were related with the reduction of the beta and gamma-secretase activities, respectively.
Amyloid Precursor Protein Secretases ; metabolism ; Amyloid beta-Peptides ; antagonists & inhibitors ; Apoptosis ; Benzyl Alcohols ; pharmacology ; Cell Line ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Humans ; Monoterpenes ; pharmacology ; Phenanthrenes ; pharmacology ; Saponins ; pharmacology

Animals ; Herbicides ; poisoning ; Humans ; Paraquat ; poisoning

Aim To investigate the antioxidant capacity of crocetin and crocin in an in vivo system.Methods Column chromatography was applied to the seperation of crocetin and crocin-1 from gardenia.Crocetin(6.25,12.5 and 25.0 mg·kg~(-1)·d~(-1)) and crocin (18.7,37.5 and 75.0 mg·kg~(-1)·d~(-1)) were orally administered to kunming mice.Then,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total antioxidant capacity(TAOC)and malondialdehyde(MDA)in mice were determined for the comparison of antioxidant activity of crocetin and crocin-1.Results Oral administration of crocetin and crocin for six weeks could enhance SOD of liver and kidney,GSH-Px of liver and TAOC of heart and kidney.In addition,it could decrease MDA of serum in mice.Conclusions The comparison of results suggests the evidence supporting the comparable antioxidant activity of crocetin and crocin.The results of the research also indicate that liver and kidney are two organs targeted for protection concerning endogenous antioxidant among various tissues.

Aim To establish a novel acute hyperuricemia mouse model and apply it to evaluate the hyporucicemia effects of Ulodesine, a purine nucleoside phosphorylase(PNP) inhibitor.Methods The mice were intraperitoneal injected inosine and subcutaneous injected Oteracil potassium to induce accumulation of uric acid, and the animal blood was collected from eyeball or vena angularis in different time points.The levels of serum uric acid were measured and determined to test whether the acute hyperuricemia mouse model were successful or not.In order to verify the hyperuricemia seen in the model was associated with the accumulation of inosine, which was converted to uric acid by action of PNP,hyporucicemia effects of Ulodesine, a PNP inhibitor, was assessed in an enzyme assay and confirmed by using the newly established model.Result Accumulation of uric acid in the blood of mouse models was observed by combined injections of intraperitoneal 200 mg·kg-1 inosine and subcutaneous 200 mg·kg-1 Oteracil potassium respectively after 1.5 h.The enzyme assay indicated that Ulodesine was a potently PNP inhibitor with IC50 of 2.293 nmol·L-1.IV injection of Ulodesine eliminated uric acid accumulations in blood of the mouse model, which was expected as the in vivo action of Ulodesine.Conclusions A novel acute hyperuricemia mouse model is established.This is a relatively easy and more effective protocol to generate the hyperuricemia in mice, which will be a useful platform to assess the anti-hyperuricemia activity of PNP-target drugs in vivo.

OBJECTIVETo evaluate clinical applicability of a novel technique that can quantify the lamivudine-resistant mutants of hepatitis B virus (HBV) in the serum of patients utilizing gene microarray technology.

METHODSThe oligonucleotide microarray was designed to detect 3 important mutational positions. Fifty-one patients who were receiving lamivudine therapy were selected as subjects. The oligonucleotide microarray and traditional sequencing were applied to detect the lamivudine resistant mutation, the monitoring lasted for 24 months. Then the clinical result was analyzed and the obtained data were compared between the two methods.

RESULTSLamivudine resistant mutation was detected in 39 percent of the patients during the 2 years period. The results of the oligonucleotide microarray technique was consistent to the results of traditional sequencing in accuracy and the miroarray was more sensitive in detection of the mixed infection.

CONCLUSIONApplication of the oligonucleotide microarray for quantitative detection of lamivudine-resistant mutation of HBV is feasible.

Antiviral Agents ; therapeutic use ; Drug Resistance, Viral ; Female ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; drug effects ; genetics ; isolation & purification ; Humans ; Lamivudine ; therapeutic use ; Male ; Mutation ; Oligonucleotide Array Sequence Analysis ; methods

Gene CaMDR1 is a member of the major facilitator superfamily (MFS),mediating multidrug resistance of Candida albicans,and can confer resistance to benomyl,fluconazole and so forth.In this review,the progress in structure and function of the protein code by gene CaMDR1 and the transcriptional regulation mechanisms of CaMDR1 are summarized.

OBJECTIVETo investigate the status of the micro-inflammation in patients undergoing maintenance hemodialysis and the effect of combined therapy with hypha cordyceps and ginkgo leaf tablet for its prevention and treatment.

METHODSSixty-five patients undergoing long-term maintenance hemodialysis were randomly assigned to the treated group (33 patients) and the control group (32 patients). Patients in both groups underwent hemodialysis regularly, meanwhile, 3 capsules of Jinshuibao (each capsule containing hypha cordyceps 0.33 g) and 1 tablet of ginkgo leaf tablet (0.23 g/tablet) were additiolally given to the patients in the treated group orally, 3 times per day, and 3 months as 1 treatment course. Their serum inflammatory factors, such as high sensitive C-reactive protein (hs-CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), which could reveal the state of microinflammation, were determined before and after treatment. And the same index was also determined in 30 healthy persons allocated in a normal control group.

RESULTSCompared with the normal control group, the serum levels of hs-CRP (4.94 +/- 2.21 mg/L), IL-6 (0.472 +/- 0.220 microg/L) and TNF-alpha (1.748 +/- 0.724 microg/L) in the treated group were higher before treatment, these indexes decreased markedly after treatment (3.28 +/- 1.28 mg/L, 0.379 +/- 0.163 microg/L, and 1.330 +/- 0.458 microg/L, respectively, all P < 0.01), but without significant change in the control group.

CONCLUSIONMicro-inflammation reaction exists popularly in patients undergoing maintenance hemodialysis, and the combined therapy with hypha cordyceps and ginkgo leaf tablet could effectively improve this kind of micro-inflammation reaction.

Adult ; Aged ; Cordyceps ; chemistry ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Ginkgo biloba ; chemistry ; Humans ; Hyphae ; chemistry ; Inflammation ; blood ; drug therapy ; etiology ; prevention & control ; Inflammation Mediators ; blood ; Male ; Middle Aged ; Plant Leaves ; chemistry ; Renal Dialysis ; adverse effects ; Tablets

OBJECTIVE: To observe the effect and the mechanism of Chrysanthemum morifolium Ramat on apoptosis of bovine aortic smooth muscle cells. METHODS: Vascular smooth muscle cells were isolated from thoracic aorta of fetal calf and cultured, then incubated with different concentration of Chrysanthemum morifolium Ramat. Apoptosis was measured by flow cytometry. SOD and MDA were measured by spectrophotometer. RESULTS: We found that: (1) the number of apoptotic cells was reduced from (4.425+/-0.624)% to (2.875+/-0.640)% in Chrysanthemum morifolium Ramat group, in a concentration dependent manner; (2) the value of SOD was increased from (1.683+/-0.149)X10(4) U/L to (2.297+/-0.230)X104 U/L and the value of MDA was reduced from(166.454+/-56.805)&mgr;mol/L to (73.068+/-27.203)&mgr;mol/L in Chrysanthemum morifolium Ramat group, also in a concentration dependent manner. CONCLUSION: Chrysanthemum morifolium Ramat can inhibit apoptosis of vascular smooth muscle cells in a concentration-dependent manner.