Objective To search for the methods preventing nephrotoxic injury of amikacin,Methods Case-control research was used in this study. There were 50 normal children in control group The urine routine, the ?2-microglobulin (?2 -M), mosmol and THP in urine and blood, the AIb, rGT and NAG in urine, the renal function and serum concentration of amikacin were determined respectively.The 43 patients with serious illness childten in study group, were divided into 2 groups (Group 1 and group 2 ). Group 1 (23 cases) was treated only with amikacin for 7 days, and group 2 (20 cases) was treated with vitaminC, vitamin E and amikacin for 7 days. Before treatment, the 3rd and 7th day during the treatment, all the items mentioncd above were examined in gtoup 1 and 2.Results The incidences of nephrotoxic injury of amikacin are 87 per cent (20/23)and 55 per cent (11/20) respectively in group 1 and 2. There is significant difference (P

Aim To prepare the monoclonal antibodies (mAbs) against human CD28 and to study its biological feature. Methods The hybridoma cell lines were obtained by fusing spleen cells of Blab/c mice that had been immunized with murine lymphoma cells transfected with full-length huaman CD28 cDNA to myeloma cells Sp2/0. Ascites were induced to produce the mAbs. The specificity and affinity of the mAb 18G8 was verified by CD28 competitive inhibitory test and FACS. Reactivities of mAb 18G8 to PBTC, U266, 8226, Jurkat and Daudi cell were studied by indirect immunofluorescence staining. mAb 18G8-inducing proliferation of peripheral blood T cells (PBTCs) was determined by [3H]thymidine incorporation test. Results Five hybridoma cell lines were obtained. mAb 18G8 secreted by one of the them, belong to mouse IgG2a. It recognized a epitope different from which recognized by the standard mAb(clone CD28.2). The Reactivitrates of the mAb 18G8 to PBTC, U266, 8266, Jurkat and Daudi cells were 70.2％ , 99.3％ , 98.6％ , 76.4％ and 1.9％ , respectively, similar with CD28.2. It was indicated that different antigen epitopes expressed on all above cells. mAb 18G8 could promote the PBTC proliferation in vitro(SI=7). It was indicated that The substitution of mAb 18G8 for B7-1 molecule could also mediate the costimulatory signals. Conclusion 18G8 is a specific and functional anti-CD28 mAb it may be of significant value in basic studies and clinical application.

The desired DNA product of KGPcd and KGP-hag was obtained from the total DNA of Porphyromonas gingivalis by PCR with two pairs of gene specific primers. The segment of KGPcd and KGP-hag (about 1.5kb and 1.6kb) was inserted into pGEM-T easy Vector. The double-stranded DNA of the postitive clone was analyzed by restriction endonuclease mapping and DNA sequenceing. The sequences of KGPcd and KGP-hag were consistent with those of the references appeared. The proteins of KGPcd and KGP-hag will be obtained for further study.

ObjectiveSLC2A9 is a novel identified urate transporter that affects serum uric acid levels. The present study is aimed to investigate rs7442295 polymorphism in intron 6 of SLC2A9 in a population of Chinese male gout or hypemricaemia subjects. MethodsA total of 268 gout patients and 288 healthy male volunteers were included. Blood pressure, body mass index(BMI), serum uric acid, glucose, lipid,urea and creatine were detected. DNA was purified from peripheral blood and the rs7442295 polymorphism was evaluated using high resolution melting ( HRM ) analysis and direct sequencing. Data were analyzed with t test or chi-square test. Results A/A and A/G genotypes were unambiguously distinguished with HRM technology. The occurrence of the homozygous type (G/G) was completely absent among the study population.The prevalence of the A/A and A/G genotype was 96.2％ and 3.8％ respectively. However, no significant differences of genotype frequencies were found in gout patients and normal subjects(x2=0.003, P=0.82; x2=0.003, P=1.00). But the serum uric acid levels in individuals with the A/G genotype[(293±100) μmol/L]were significantly lower than those with the A/A genotype[(392±133) μmol/L](t=2.426, P＜0.01 ). The A/G genotype frequency was significantly higher in the low-uric acid group than in the high uric-acid group (x2=6.279, P=0.01 ). Genotyping based on HRM was fully concordant with sequencing. Conclusion The polymorphism rs7442295 in SLC2A9 may be a genetic marker to assess risk of hyperuricemia among Chinese male Hart population. HRM is a simple, fast, reliable and close-tube technology for genotyping.

OBJECTIVETo study the association between endoplasmic reticulum stress (ERS) pathway mediated by inositol-requiring kinase 1 (IRE1) and the apoptosis of type II alveolar epithelial cells (AECIIs) exposed to hyperoxia.

METHODSThe primarily cultured AECIIs from preterm rats were devided into an air group and a hyperoxia group. The model of hyperoxia-induced cell injury was established. The cells were harvested at 24, 48, and 72 hours after hyperoxia exposure. An inverted phase-contrast microscope was used to observe morphological changes of the cells. Annexin V/PI double staining flow cytometry was performed to measure cell apoptosis. RT-PCR and Western blot were used to measure the mRNA and protein expression of glucose-regulated protein 78 (GRP78), IRE1, X-box binding protein-1 (XBP-1), and C/EBP homologous protein (CHOP). An immunofluorescence assay was performed to measure the expression of CHOP.

RESULTSOver the time of hyperoxia exposure, the hyperoxia group showed irregular spreading and vacuolization of AECIIs. Compared with the air group, the hyperoxia group showed a significantly increased apoptosis rate of AECIIs and significantly increased mRNA and protein expression of GRP78, IRE1, XBP1, and CHOP compared at all time points (P<0.05). The hyperoxia group had significantly greater fluorescence intensity of CHOP than the air group at all time points. In the hyperoxia group, the protein expression of CHOP was positively correlated with the apoptosis rate of AECIIs and the protein expression of IRE1 and XBP1 (r=0.97, 0.85, and 0.88 respectively; P<0.05).

CONCLUSIONSHyperoxia induces apoptosis of AECIIs possibly through activating the IRE1-XBP1-CHOP pathway.

Animals ; Apoptosis ; Cells, Cultured ; Endoplasmic Reticulum Stress ; physiology ; Endoribonucleases ; physiology ; Epithelial Cells ; physiology ; Female ; Hyperoxia ; metabolism ; pathology ; Multienzyme Complexes ; physiology ; Protein-Serine-Threonine Kinases ; physiology ; Pulmonary Alveoli ; pathology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor CHOP ; physiology ; X-Box Binding Protein 1 ; physiology

Scutellarin is the main effective constituent of breviscapine, a flavonoid mixture isolated from the dried whole plant of Erigeron breviscapus (Vant.) Hand-Mazz, and valsartan is used as an antihypertensive drug. These two drugs have already been clinically used together to treat diabetic nephropathy (DN) in China, and the combined medications showed some enhanced protection against DN. The aim of this study is to investigate the potential pharmacokinetic interaction between scutellarin and valsartan in rats. Breviscapine injection (20 mg x kg(-1), i.v.) and valsartan (15 mg x kg-, i.g.), either alone or together were given to 18 male Sprague-Dawley rats. Concentrations of scutellarin and valsartan were quantified by HPLC, and pharmacokinetic parameters were calculated by non-compartmental methods. We found that the pharmacokinetic parameters of scutellarin altered significantly after co-administration of oral valsartan. The plasma clearance (CL(p)) and the bile clearance (CL(b)) of scutellarin were reduced significantly in the presence of valsartan. After oral administration of valsartan with or without intravenous scutellarin, however, the pharmacokinetic parameters of valsartan were comparable. In conclusion, our data suggests that the concurrent use of valsartan reduces the biliary excretion of scutellarin, and this may be due to the inhibitory effect of valsartan on the biliary excretion of scutellarin mediated by Mrp2 (Multidrug resistance-associated protein 2).
Administration, Intravenous ; Administration, Oral ; Animals ; Antihypertensive Agents ; administration & dosage ; blood ; pharmacokinetics ; Apigenin ; administration & dosage ; blood ; isolation & purification ; pharmacokinetics ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Drug Interactions ; Erigeron ; chemistry ; Glucuronates ; administration & dosage ; blood ; isolation & purification ; pharmacokinetics ; Male ; Metabolic Clearance Rate ; Multidrug Resistance-Associated Proteins ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Valsartan ; administration & dosage ; blood ; pharmacokinetics

AIMTo study on synthesis and antibacterial activity evaluation of polyheterocycles.

METHODSThe condensation of 4-amino-3-pyridin-3-yl-4H-[1,2,4] triazole-5-thiol with 2-chloromethyl-5-substituted phenyl-[1,3,4] oxadiazoles gave the corresponding title heterocycle amines, and the in vitro antibacterial activity was primarily evaluated by the method of cup-plate diffusion solution.

RESULTSTwelve novel compounds were synthesized, and their structures were confirmed by IR, 1H NMR, MS and element analysis. Biological screening results demonstrated that most of the compounds prepared showed good antibacterial activity.

CONCLUSIONOxadiazoles incorporting pyridyl triazole ring may be a pharmacophor structure in the molecule for developing antibacterial candidate drugs.

Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Escherichia coli ; drug effects ; Oxadiazoles ; chemical synthesis ; chemistry ; pharmacology ; Proteus vulgaris ; drug effects ; Staphylococcus aureus ; drug effects ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

Objective To explore the clinical value of the level of granzyme B and perforin mRNA in urine for the diagnosis of renal transplantation patients with delayed graft function (DGF). Methods Twenty-four cases of renal transplantation patients with DGF were included in this study. Seventy-three u-rine specimens were obtained from these patients who received graft biopsies. Among the 24 cases, ureteral obstruction occurred in 2 cases, vascular thrombosis in 1 case, acute CsA intoxication in 3 cases, acute tubu-lar necrosis (ATN) in 7 cases, ATN complicating borderline change in 2 cases, ATN complicating acute re-jection (AR) in 3 cases, AR in 6 cases. Total RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B gene were measured with the quantitative polymerase-chain-reaction assay-(RT-PCR). SPSS13.0 software was used for data analysis. Levels of mRNA were log-transformed before analysis. Results The levels of perforin and granzyme B mRNA in u-rine among the ureteral obstruction group, vascular thrombosis group, acute CsA intoxication group and ATN group were very low. There was no significant difference among these groups (P>0.05). However,among the ATN complicating borderline change group 1.22, 0. 97 fg/μg, ATN complicating AR group (1.20±0.39), (1.07±0.30)fg/μg, and AR group(11.13±0. 33), (1.01±0.19)fg/μg, the levels were increased significantly(P<0.001). Conclusion Measurement of mRNA encoding the cytotoxic proteins perforin and granzyme B gene in urinary cells in renal transplantation patients with DGF could be helpful to etiological diagnosis of DGF, and might be used as an index for the appropriate management of the borderline change.

Objective To explore the association between polymorphisms of interferon-gamma gene intron 1 at position+874 (IFN-γ+874) gene and the susceptibility of HBV and/or HCV infection with different clinical outcomes. Methods IFN-γ+874 gene SNP were detected in 277 subjects including 79 chronic HBV/HCV coinfections,69 individuals only with HBV infection,55 individuals only with HCV infection and 74 controls,by sequence specific primers-PCR (SSP-PCR). Hepatocellular injury as suggested by alanine aminotransferase (ALT) was detected by Beckman LX-20. The status of viral particles in serum was determined by RT-nPCR. The possible association of the polymorphism of IFN-γ+874 with the susceptibility of HBV and/or HCV infection and the outcome of these infections were analyzed. Results (1) IFN-γ+874 AA frequency in individuals with chronic HBV,HCV,HBV/HCV coinfections were significant higher than that in controls (X~2=16.15,P=0.01); OR (95% CI) of IFNγ+874 AA in chronic infection with HBV,HCV,HBV/HCV coinfections appeared to be 2.70 (1.24-5.92),3.22 (1.43-7.25) and 4.02 (1.88-8.55) compared with + 874 TA. No significant differences were found among HBV,HCV,HBV/HCV coinfections (X~2=1.97,P=0.73). There were no significant association of IFN-γ +874 A/T allele frequency with HBV and/or with HCV infection (X~2=4.87,P=0.18). (2)The clinical outcomes of mild chronic hepatitis (CH),moderate/severe CH and cirrhosis with HBV and/or HCV infection were associated with IFN-γ+874 AA [X~2＝14.17,P＝0.03;OR＝3.09(1.51-6.33),3.85 (1.70-8.70),3.14 (1.08-9.17)]. No significant relationships were found between IFN-γ+874 A/T allele frequency and the clinical outcome of HBV/HCV infection (X~2＝6.07,P＝0.11). (3)There were no significant associations of IFN-γ+874 genotype/allele frequency with HCV duplication (X~2=2.36,P=0.31). (4) There were no significant associations of IFN-γ+874 genotype/allele frequency with abnormal ALT (X~2=0.15,P=0.93). Conclusion These results suggested that polymorphisms in the IFN-γ +874 had some influence on chronic HCV and/or HBV infection,and on the outcome of HCV and/or HBV infections. IFN-γ+874 AA genotype and T allele were possible risk to chronic HBV and/or HCV infections and to the outcomes of HBV and/or HCV infection. However,IFN-γ+874 TA genotype might serve as possible protective factors to them.

Objective To study the relationship between polymorphisms in interleukin-2gene at position-330 (IL-2-330) and the clinical outcome of hepatitis B virus (HBV) and/or hepatitis C virus (HCV) infection. Methods 277 subjects were recruited including 79 chronic HCV co-HBV infection, 55 chronic HCV infection, 69 chronic HBV infection and 74 controls. Single nucleotide polymorphisms of IL-2-330 was investigated by restricted fragment long polymorphism-PCR (RFLP-PCR). Hepatocellular injury, as revealed by alanine aminotransferase (ALT) was detected by Beckman LX-20 analyzer. The presence of hepatitis C viral particles in serum was determined by RT-nPCR. Results ( 1 ) IL-2-330 polymorphisms showed close association with persistent HBV and/or HCV infection. IL-2-330 TT was associated with an increased risk, but IL-2-330 GG with a reduced risk of persistent HBV and/or HCV infection (χ2=14.24, P=0.03 ) with ORs (95%CI) as 7.14(2.13-23.81 ), 3.46 (1.17-10.02) and 2.93 (1.15-7.46) respectively. However,IL-2-330 TT/GG did not significantly differ between patients with HBV and/or HCV infection (χ2=2.09, P=0.72). IL-2-330 T allele was associated with an increased risk, but the -330G allele was associated with a reduced risk of chronic HBV/HCV infection (χ2=12.33,P=0.01),with ORs (95% CI) as 2.26 (1.39-3.69) , 1.82 ( 1.09-3.03 ) and 1.73 ( 1.10-2.73 ) respectively. (2) IL-2-330polymorphisms showed significant association with the outcome of HBV and HCV infection ( χ2=13.52, P=0.04). IL-2-330 TT was associated with an increased risk, but-330 GG with a reduced risk of mild CH, moderate/severe CH, and cirrhosis. The ORs (95%CI) appeared to be 3.33(1.75-6.32), 3.31 (1.75-6.26), 11.23 (3.09-40.76) respectively. IL-2-330 T allele was associated with an increased risk, but the -330 G allele was associated with a reduced risk of mild CH, moderate/severe CH and cirrhosis (χ2= 12.32, P=0.01 ), with ORs as 1.86(1.32-2.63), 1.71 (1.27-2.31) and 2.77(1.57-4.89) respectively. (3) The polymorphisms of IL-2-330 showed no association with HCV RNA replication (χ2=0.83, P=0.66; χ2=0.20, P=0.66). The polymorphisms of IL-2-330 were not significantly associated with abnormal ALT ( χ2= 1.10, P=0.58; χ2=0.08, P=0.78). Conclusion These results suggested that IL-2-330 TT/T was associated with an increased risk, but IL-2-330GG/G was associated with reduced risk of persistent HBV and/or HCV infection, and with the development of mild CH,moderated/severe CH,and cirrhosis.