Background Oxidative stress is thought to be responsible to diabetes-complicated cataract.Our previous study demonstrated that as an iron chelator,deferiprone can protect lens from oxidative damage.Objective This further study aimed to investigate the role of deferiprone on the formation of diabetic-complicated cataract.Methods Forty 6-week-old Wistar rats were included in the study and randomized into 4 groups.Eight of them were used as the normal control group.Diabetes mellitus animal models were established in 22 rats by the carbonhydratediet and fat diet and the intraperitoneal injection of 40 mg/kg streptozocin (STZ).The deferiprone of 50 mg and 100 mg were intragastrically given in 8 model rats respectively after 3 days once a day for 8 weeks.The opacification of lenses was examined under the slit lamp weekly after treatment.The animals were sacrificed and the lenses were obtained at the eighth week of deferiprone injection.The concentrations of water-soluble protein ( WSP),urine-soluble protein (USP) and alkali-soluble protein (ASP) in rat lens suspension were detected by Bradford method.The super oxide dimutese (SOD),malondialdehyde (MDA) and glutathione (GSH) were determined spectrometically using xanthine oxidase,thiobarbituric acid,dithio bis-nitrobenzoic acid.Results No evidently differences were found in the content of the WSP,USP and ASP among the these groups( F=1.73,0.18,0.09,P＞0.05).The contents of MDA in 50 mg deferiprone group and 100 mg deferiprone group were ( 1.05 ± 0.10 ) mmol/g and ( 1.05 ± 0.22 ) mmol/g respectively,showing a significant decline in comparison with diabetic model group (P＜0.05).The SOD and GSH contents in lens were (321.29±16.57) U/mg,(322.07±22.16) U/mg and (7.83±0.65 ) mg/g,(7.70±0.77 ) mg/g respectively in 50 mg deferiprone group and 100 mg deferiprone group and were considerably elevated in comparison with ( 298.70± 14.69 ) U/mg and ( 5.47 ± 1.01 ) mg/g of diabetic model groups ( P＜0.05 ).No significant differences were found in the indexes mentioned above between 50 mg and 100 mg deferiprone groups(P＞0.05).Conclusions Deferiprone can reduce oxidative stress and improve the energy metabolism of the lens in diabetic rats.

OBJECTIVETo investigate the effect of inhaled nitric oxide (NO) on surfactant protein A (SP-A) and mannose binding ability (MBA) in neonatal rats with hyperoxia-induced lung injury.

METHODSSixty-four neonatal rats were randomly exposed to room air (Control group), >95% oxygen for 6 days (Hyperoxia group), 10 ppm NO for 24 hrs (NO group), and >95% oxygen for 6 days along with 10 ppm NO for 24 hrs (Hyperoxia + NO group). After 2 and 6 days of exposure, the lung pathologic changes, gene and protein expressions of SP-A and MBA were measured.

RESULTSThe rats from the Hyperoxia group presented with obvious lung injuries. The SP-A expressions of mRNA (0.81 +/- 0.04 vs 1.53 +/- 0.25) and protein (59.45 +/- 18.37 vs 89.77 +/- 16.41) in the Hyperoxia group decreased significantly 2 days after exposure but increased significantly 6 days after exposure (SP-A mRNA 0.81 +/- 0.02 vs 0.63 +/- 0.03; SP-A protein 93.57 +/- 13.71 vs 47.73 +/- 21.69) compared with those of the Control group (P < 0.05). NO treatment alleviated the hyperoxia-induced pathologic injuries 2 days after exposure. The SP-A mRNA expression (0.55 +/- 0.91) in the Hyperoxia + NO group was significantly reduced as compared to both the Control and Hyperoxia groups (P < 0.05), and the SP-A protein expression (55.12 +/- 17.53) in the Hyperoxia + NO group was noticeably lower than that of the Control group (P < 0.01) 2 days after exposure. The SP-A protein expression in the Hyperoxia + NO group (67.33 +/- 18.59) was significantly lower than that of the Hyperoxia group 6 days after exposure (P < 0.05). Two days after exposure, the NO group had significantly higher MBA than the Control group (0.821 +/- 0.133 vs 0.58 +/- 0.158); the Hyperoxia + NO group had significantly higher MBA than the Hyperoxia group (0.43 +/- 0.175 vs 0.738 +/- 0.141) (P < 0.05).

CONCLUSIONSInhaled low dose NO may decrease SP-A protein expression and increase MBA of the lung tissue. This lessens the pathologic lung injury in neonatal rats with hyperoxia.

Administration, Inhalation ; Animals ; Animals, Newborn ; Hyperoxia ; pathology ; Lung ; drug effects ; metabolism ; pathology ; Mannose ; metabolism ; Nitric Oxide ; administration & dosage ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To discover the effects of the Prescription Administrative Policy in force on the quality of the prescriptions in a tertiary hospital in 2007. Methods The prescriptions of 400 cases were sampled systematically for evaluation, and 60 patients were interviewed. Results The average eligibility rate of the prescriptions was but 37. 2% in this hospital, which was mainly plagued by incompleteness, nonstandard and irrationality found in prescriptions. Implementation of the Policy contributed to a significant improvement of some indicators. For example, the eligibility rate increased by 12. 2% (P=0. 004) ,the percentage of the use of antimicrobial agents dropped significantly (P=0. 001),and the percentage of generic names used rose significantly (P = 0. 000). Conclusions The implementation of the Policy is highly positive for prescription quality.

Benzene ; analysis ; Chemical Industry ; Chromatography, Gas ; methods ; Humans ; Occupational Exposure ; analysis ; Toluene ; analysis ; Trichloroethylene ; analysis ; Xylenes ; analysis

AIM:To construct a hAR and GFP fusion gene vector and to observe the AR-GFP gene expression in Hek293 cells. METHODS: A recombined vector pcDNA3.1/myc-HisA-AR-GFP (pH-AG) was constructed by gene engineering technique. The recombined vector was transfected into Hek293 cells using calcium phosphate. RESULTS: AR-GFP fusion protein was successfully expressed in Hek293 cells without biologic activity, which was confirmed by fluorescence microscopy and Western blotting. CONCLUSION: The Hek293 cells transfected by AR-GFP fusion gene can express its protein successfully. However, it is not a cellular model for ARI screening.

Objective:Aldose reductase,involved in the pathogenesis of diabetic complications,was recombinated with an adeno associated virus vector pSNAV2.0,and it was transfected into human embryonic kidney 293(HEK 293)cells.The gene engineering produced AR would be used as a target protein to screen aldose reductase inhibitors.Restriction endonuclease digestion and ligation procedures were performed to construct the AR expression plasmid vector pSNAV-hAR.Methods:After confirmation the recombinant plasmid by PCR,restriction endonuclease digestion,and DNA sequencing,pSNAV-hAR was transfected into HEK293 cells.Western blot and immunofluorescence analysis were performed to detect the expression of AR and its enzyme activity.Results:The results of a series of analysis including AR activity assay,Western blot and immunofluorescence analysis shown the expressed protein mediated by the adeno associated virus vector transfecting HEK 293 cells,was functional AR.The traditional aldose reductase inhibitors,Sobinil and Zopolrestat,were used to test and verify the constructed cell model.Conclusion:The established AR expression model can be used in mechanismresearch of activation of polyol pathway on diabetic complications and screening potential aldose reductase inhibitors.

Objective:To systematically evaluate the risks of anti-TNF-αtreatment-associated infection, severe infection and tuberculosis in rheumatoid arthritis (RA) patients, and to reduce the infection incidences associated with anti-TNF-αtherapy. Methods:We used Meta analysis to systematically review randomized controlled trials on anti-TNF-αtreatment associated risks of infecion, severe infection and tuberculosis in AR patients.Results:Although no statistically significant differences were detected in TB risk between anit-TNF-αtreatment and the control group (0.5%vs 0.07%;P=0.27, OR=1.85, 95%CI:0.62-5.52), there still existed a clinically obvious elevation of TB risk in monoclonal anti-TNF-αtreatment, which was illustrated by the results that no TB case was reported in the etanercept group, but 11 TBs in 2050 infliximab-treated cases, and 3 TBs in 722 adalimumab-treated cases. The total infection and severe infection risks were also signiifcantly higher in patients receiving anti-TNF-αtreatment (P<0.05). Subanalysis revealed that etanercept showed no signiifcantly higher infection or severe infection risk than control group (P>0.05), while both kinds of monoclonal antibodies of TNF-αblockers showed a signiifcantly elevated infection or severe infection risks (P<0.05). High doses of anti-TNF-αtreatment were associated with statistically increased risks of severe infection (6.0%vs 2.8%, P=0.04, OR=1.68, 95%CI:1.02-2.78). Conclusion:The TB risk of anti-TNF-αtreatment deserves close attention, especially in places with high rate of BCG vaccination and MTb infection. Monoclonal anti-TNF-αtreatment brings higher risks of infection and severe infection than soluble TNF-αreceptor.

Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines (PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990) was detected by the methylationspecific PCR (MSP) method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine (5-Aza-dC) treatment, a kind of DNA methyltransferase (DNMT) inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression (RQ) were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.

To investigate the effect of Kozak sequence (+4A or +4G) on expression of green fluorescent protein (GFP) gene in HEK293 cells. The eukaryotic expression vectors containing GFP gene with different Kozak sequence (+4A or +4G) were constructed by classic DNA recombination methods, including PCR, enzyme digestion, ligation, transformation, identification, et al. Two different Kozak sequences (+4A or +4G) were obtained through PCR with different mutagenic primers. The right recombinant plasmids pHGFP-A and pHGFP-G were transfected into HEK293 cells by liposome-mediated gene transfer method. The expression level of GFP was observed by fluorescent microscope, flow cytometry and Western blot. The flow cytometry revealed that the expression levels of GFP fluorescence in pHGFP-A and pHGFP-G transfected cells were about 15% and 45%, respectively. Western blot showed the specific bands of about 27 kD (GFP) both in pHGFP-G and pHGFP-A sample lanes; and the GFP expression density of pHGFP-G was about 3.87-fold as that of pHGFP-A by ImageJ software analysis. These results indicated that the +4G in Kozak sequence (when -3 site is purine base pair) plays an important role in GFP protein translation, which enhances the GFP expression up to 4-fold in HEK293 cells.
Actins ; biosynthesis ; genetics ; Cell Line ; Cloning, Molecular ; Gene Expression Regulation ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Protein Processing, Post-Translational ; RNA, Messenger ; metabolism ; Recombination, Genetic ; Transfection

OBJECTIVETo study the relation between body mass index (BMI) and percentage body fat (PBF) in Chinese adolescent, and to compare the prevalence of overweight and obesity using BMI or PBF standards.

METHODSBMI from 757 girls with an average aged of 10.1 years in the rural areas, 165 girls average aged 13.5 years in suburbs and 172 boys average aged 13.7 years in suburb of Beijing, were measured. Their body compositions were also measured by dual-energy X-ray absorptiometry (DEXA).

RESULTSBMI was found closely correlated with PBF in each age group of rural and suburb girls and suburb boys with the correlation coefficient(r) of 0.59 - 0.83. When age, height and pubertal development were controlled, r became 0.54 - 0.88. The prevalence rates of obesity in rural girls, suburb girls and suburb boys were 33.1%, 21.8% and 21.5%, when PBF cutoff values (girls: PBF >or= 35%, boys: PBF >or= 25%) were used. However, the rates became 2.4%, 3.0% and 4.0% when BMI cutoff values of International Obesity Task Force (IOTF) were used. Compared with PBF cutoff values for obesity, the IOTF recommended BMI cutoff values had relatively lower sensitivity (7.3% - 18.9%) and higher specificity (100%).

CONCLUSIONBMI correlated well with PBF in Beijing adolescent. IOTF-BMI cutoff values showed low sensitivity and high specificity to Chinese adolescent which might be suitable for identifying obesity but not for the purpose of screening.

Adolescent ; Adolescent Development ; physiology ; Age Factors ; Body Composition ; physiology ; Body Mass Index ; Child ; China ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Multivariate Analysis ; Obesity ; diagnosis ; epidemiology ; Rural Population ; statistics & numerical data ; Sex Factors ; Suburban Population ; statistics & numerical data ; Urban Population ; statistics & numerical data