Objective To study the impact of sodium arsenite(NaAsO2) exposure on melanoma cells A375 (hereinafter referred to as the A375) and G361 (hereinafter referred to as the G361) pigment production and tyrosinase (TYR) activity and the differences of pigment metabolism capacity between the cell lines.Methods A375 and G361 cells were exposed to sodium arsenite at concentrations of 0.0(control),0.1 and 1.0 μmol/L for 72hours.Cell viability was measured by Alamar Blue assay.Melanin levels and TYR activity were measured at the same time.Results After exposure for 72 hours,the cells of 0.1 μmol/L dose groups of both of the two cell lines [A375:(103.32 + 1.26)％; G361:(104.10 + 1.76)％] showed a slightincrease of proliferation without significant differences compared with those of the control[A375:(100.00 ± 1.08)％; G361:(100.00 + 1.79)％,all P ＜ 0.05] ;while cell viability of the 1.0 μmol/L dose group of both of the two cell lines[A375:(75.32 ± 1.59)％; G361:(78.26 ± 2.10)％] were significantly lower than those of the control (all P ＜ 0.05).Melanin levels of G361 cell line [(7.19 ± 0.35),(7.34 ± 0.83),(8.19 ± 0.86)pg/cell] were significantly higher than that of A375[(4.35 ± 0.72),(4.54 ± 0.01),(4.60 + 0.59)pg/cell,all P ＜ 0.05] in all the three groups.TYR activity of G361 cell line [(54.13 ± 1.21),(54.56 ± 0.21),(56.25 ± 0.85)Bq] were also markedly higher than that of A375 cell[(42.00 ±0.21),(42.90 ± 0.54),(42.91 ± 0.01)Bq,all P ＜ 0.05] in all the three groups.The melanin levels and TYR activities of both of the two cells lines showed an increase tendency along with increased doses of arsenic exposure,but without significant differences when compared with those of the three groups (all P ＞ 0.05).Conclusions Arsenic related pigment disorder may be associated with increased melanin levels and TYR activities induced by arsenic exposure; individual difference of pigment metabolism may be associated with different basal melanin levels and TYR activity between different individuals.

The initial study of the tumour DNA fingerprints using MYO minisatellite DNA probe wascarried out,and then,by means of DNA recombinant techniques,the fragment of MYO min-isatellite DNA probe obtained from plasmid pUC19-MYO was inserted into plasmid pGEM-4Z containing RNA polymerase promotor,thus a sub-clone refferred as pGEM-4Z-MYOwas constucted.That made an offer of the conditions of preparing RNA probe in order to in-cerase the sensitivities of DNA fingerprinting and laid a foundation for raised the efficiency of de-tecting the polymorphism of the minisatellite DNA.

AIM:To observe the effects on human keratocytes by cationic liposome LipofectamineTM 2000(LF2000).to investigate the efficiency and safe range applied in human keratocytes,and establish basis for gene therapy of human keratocytes.METHODS: Human keratocytes cultured in vivo within 3 to 5 passages were used in experiment after being identified.The effects on proliferation of cultured human keratocytes by LF2000 with different concentrations and time were evaluated By MTT:the effects of LF2000 on the survival rate and its relation with 5,10,20,40.80mg/L concentration and time were detected by trypan blue staining.related with concentration and time.The cellular proliferation and survival rate declined when concentration of LF2000 was above certain level,and this effect increased as time became longer.LF2000 had no effect with concentration under 40mg/L for 24 hours. CONCLUSION:LF2000 did ont cause cytotoxicity during a concentration range"tested",and it is hoped to play an important role in gene therapy of human keratocytes.

A total of 906 subjects with no history of diabetes who had undergone coronary angiography were included in this study and categorized into four groups according to the level of fasting plasma glucose (FPG):≤5.5 mmoL/L,5.6-6.0 mmol/L,6.1-6.9 mmoL/L,and ≥ 7.0 mmol/L.Significant coronary artery disease (CAD) was defined as ≥ 50％ reduction of lumen diameter at least in one major coronary artery.The severity of coronary atherosclerosis was defined by the Gensini score.The clinical data,laboratory indexes,and coronary angiography results were compared among various groups.The risk factors for the prevalence and severity of angiographic CAD were analyzed.The results showed that the prevalence of angiographic CAD,the number of diseased vessels,and the Gensini score were increasing with increasing FPG levels among four groups (P＜0.05 or P＜0.01).The FPG level was significantly correlated with angiographic CAD (P =0.004) and the Gensini score (P =0.010),suggesting that FPG was an independent risk factor for the prevalence and severity of angiographic CAD.

Colitis, Ulcerative ; Humans ; Infant, Newborn ; Male

Objective To study the effect of low-dose FK778 in preventing chronic renal al- lograft rejection in rats.Methods The rat model of chronic renal allograft rejection was established by using micro-surgery technique.The recipients were divided into two groups.The recipients in the study group were treated with FK778 at a dose of 5mg?kg~(-1)?d~(-1)dissolved in carboxymethylcellulose by means of gavage and the controls were treated with carboxymethylcellulose.Urinary protein con- centrations were measured every 4 weeks for 24 weeks.On 24th week after operation,the rats were killed and the kidney grafts were taken out for histological and immunohistological examinations as well as quantitative real-time RT-PCR analysis.Results After 24 weeks of treatment,proteinuria, the severity of chronic rejection,glomeruIosclerosicytes and monocytes/macrophages in the study group were significantly milder than in control group.And the expression of TGF-?mRNA and PDGF-B mRNA was significantly reduced in the study group as compared with that in the control group.Conclusion Low-dose of FK778 might prevent the rats from chronic renal allograft rejection.

Humans ; Infant, Newborn ; Male ; Polycystic Kidney Diseases ; diagnosis

·AIM: To study the tear function changes in patients with pterygium before and after pterygium transposition.·METHODS: Twenty eyes of 20 patients were enrolled in this study.Schirmer I test and tear break-up time (BUT) were evaluated in patients before and after pterygium transposition.·RESULTS: There was no significant difference in the results of Schirmer I test before and after the surgery.The results of BUT were not significantly different before and up to 4 weeks after surgery. However, BUT prolonged significantly 6 weeks after surgery (P<0.05).·CONCLUSION: Pterygium transposition can improve the tear film function in patients with pterygium.

Background The main cause of X linked juvenile retinoschisis is mutation of RS1 gene.The phenotype of X linked juvenile retinoschisis is associated with the mutation types of RS1 gene.However,the relationship of genotype and phenotype of X linked juvenile retinoschisis is unclear.Objective The present study was to survey the clinical phenotype of X-linked juvenile retinoschisis in twelve Chinese families with eleven different mutations in the XLRS1 gene. Methods Complete ophthalmic examinations with slit lamp biomicroscopy,fundus examination and Dhotography were carried out in 28 affected males.Ganzfeld electroretinography (ERG),fundus fluorescein angiography,A and B-scan standardized echography and optical coherence tomography(OCT)were also performed in some patients.The coding regions of the XLRS1 gene that encodes retinoschisin were amplified by polymerase chain reaction(PCR)and analyzed by the single strand conformation polymorphism(SSCP)assay.The RS1 gene mutations were determined by direct sequencing in an automated sequencer.Written informed consent wasobtained prior to the survey. Results The 28 affected males showed a typical foveal schisis with or without peripheral retinoschisis.The typical response to white single flash ERG was seen with a reduction of the b-wave amplitude and a relative preservation of the a-wave amplitude.causing a reduced b/a ratio in the male patients.A total of eleven different XLRS1 mutations in 12 families were identified,four of these mutations,including one frameshift mutaion(22 del T)of exon 1,Asp145His,Arg156Gly and Trp163X mutations of exon 5,were first described in this survey.One non-disease-related polymorphism(NSP),or the 576C to T(Pro192Pro)change of exon 6 was also newly reported herein.In the families with a frameshift(22 del T)mutation of exon 1,a splice donor site mutation(IVS1+2T

ObjectiveTo study the protective effects of tert-butylhydroquinone(tBHQ) on sodium arsenite (NaAsO2)-induced cytotoxicity and oxidative injuries. Methods Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L]for 24 h, and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control),30, 40, 50, 60 μmol/L] for another 24 h, and Alamar blue reduction rates were used to evaluate cell viability,the results were expressed as the relative ratio of Alamar blue reduction rates between the experimental group and the control group. On the other hand, Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L] for24 h,and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control), 40, 50 μmol/L] for another 24 h,and the levels of cellular reactive oxygen species(ROS) were detected by staining cells with 2',7'-dichlorofluorescin diacetate(DCFH-DA), the results were expressed as the relative ratio of mean fluorescence intensity between the experimental group and the control group. ResultsCell viability decreased dramatically by treatment with NaAsO2(30, 40, 50, 60 μmol/L), while relieved to some extent by pretreatment with 5, 25 μmol/L tBHQ, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant(F =566.57, 55.09, 14.50,all P ＜ 0.05) ; the cell viability of NaAsO2(30, 40, 50, 60 μmol/L) pretreated with tBHQ(5, 25 mol/L) were 0.75 ±0.02, 0.70 ± 0.04, 0.59 ± 0.03, 0.43 ± 0.03 and 0.75 ± 0.02, 0.73 ± 0.03, 0.65 ± 0.02, 0.50 ± 0.02, respectively,all significantly higher than corresponding NaAsO2 alone groups(0.70 ± 0.03, 0.64 ± 0.03, 0.43 ± 0.03, 0.33 ±0.01, all P ＜ 0.05), the cell viability of NaAsO2(50, 60 μmol/L) pretreated with 25 μmol/L tBHQ was higher than corresponding 5 μmol/L tBHQ pretreatment groups(all P ＜ 0.05). On the other hand, 40, 50 μmol/L of NaAsO2 significantly induced hepatocellular ROS generation, while tBHQ(5, 25 μ mol/L) pretreatment significantly decreased NaAsO2-induced intracellular ROS levels, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant (F =181.78, 60.55, 4.93, all P ＜ 0.05) ; the ROS levels of NaAsO2(40, 50 μ mol/L) pretreated with tBHQ(5, 25 μmol/L) were 1.87 ± 0.09, 1.80 ± 0.07 and 1.36 ± 0.11, 1.44 ± 0.12,all significantly decreased than corresponding NaAsO2 alone groups(2.30 ± 0.18, 2.18 ± 0.17, all P ＜ 0.05),the ROS levels of NaAsO2(40, 50 μmol/L) pretreated with 25 μmol/L tBHQ decreased than corresponding 5 μmol/L tBHQ pretreatment groups (all P ＜ 0.05). ConclusiontBHQ has a certain antagonism on arsenic induced cytotoxicity and oxidative injuries.