Background Ultraviolet irradiation promotes cellular apoptosis by affecting the mitochondrial transmembrane potential,including human lens epithelial cells (LECs).Gene associated with retinoid-interferoninduced mortality-19 (GRIM-19),a cell death regulatory protein,is essential for the assembly and function of mitochondrial complex Ⅰ.However,whether LECs apoptosis induced by ultraviolet irradiation is related to GRIM-19 is still unclear.Objective The purpose of this study was to investigate the relationship between the apoptosis of human LECs caused by ultraviolet with GRIM-19 expression in vitro.Methods Human LEC line(SRA01/04)was cultured in α-MEM containing 10％ fetal bovine serum.The cells were exposed to ultraviolet ray at doses of 0,30,60,90,120 or 150 mJ/cm2 when cell growth reached the logarithmic phase and 80％ confluency.The rate of apoptosis of the cells was assayed using flow cytometry,and the level of expression and relative amount of GRIM-19 protein (GRIM-19/β-actin) were detected by Western blot.The relationship between apoptosis and the GRIM-19/β-actin value among the different treatment groups was compared using One-way ANOVA,and the correlation of LECs apoptosis rate and GRIM-19 expression level was assessed by Pearson linear analysis.Results A significant difference was found in the apoptosis rate among the different treatment groups(F=149.32,P＜0.01).Compared with the 0 mJ/cm2 ultraviolet irradiation group,the apoptosis rate of LECs was significantly increased in the 60,90,120 and 150 mJ/cm2 ultraviolet irradiation groups (q =17.02,-25.20,-29.41,-8.61,P ＜ 0.01).The expression of the GRIM-19 protein in the LECs suspension was enhanced by ultraviolet irradiation at 60,90,120 and 150 mJ/cm2.The relative expression of the GRIM-19 protein (GRIM-19/β-actin) was significantly different among the various groups (F=6.87,P＜0.05),and the GRIM-19/β-actin values in the 60,90,120,150 mJ/cm2 ultraviolet irradiation groups were elevated in comparison with the un-irradiated group(2.01±0.76,2.98± 1.80,3.97± 1.61,2.42± 1.28 vs.0.56±0.23),which showed statistically significant differences (q =4.12,-5.04,-7.09,-3.85,P ＜ 0.01).In addition,a positive correlation was seen between the rate of apoptosis and the expression of the GRIM-19 protein(r=0.71,P＜0.01).Conclusions GRIM-19 is expressed in normal human LECs.The apoptosis of human LECs accompanies the up-regulation of GRIM-19.The expression of GRIM-19 in LECs increases with ultraviolet irradiation in a doseindependent manner.

Background Epithelial-myofibroblast transition (EMT) of human lens epithelial cells (LECs) induced by transforming growth factor-β2 (TGF-β2) is the main mechanism in the pathogenesis of posterior capsular opacification(PCO).Seeking an effective drug capable of inhibiting this process is important for the prevention and treatment of PCO.Objective The purpose of this study was to investigate the inhibitory effect of rapamycin (RAPA)on the proliferation of human LECs and TGF-β2-induced EMT.Methods Human LEC strain(SRA01/04)was cultured in DMEM with high glucose and 10％ fetal bovine serum.The cells were consequently cultured in serumfree DMEM with 5 mg/L TGF-β2,TGF-β2+10 mg/L RAPA,TGF-β2 + 100 mg/L RAPA,TGF-β2 + 1000 mg/L RAPA or TGF-β2 +10 000 mg/L RAPA for 72 hours,and SRA01/04 cultured in serum-free DMEM were used as control.The proliferation rate(A490)of SRA01/04 in the different groups was detected using the MTT assay and the rate of inhibition of RAPA was calculated.The expressions of the α-smooth muscle actin(α-SMA) and E-cadherin(E-cad)mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.The changes in the expression of α-SMA and E-cad in SRA01/04 were evaluated by Western blot 24,48 and 72 hours after TGF-β2 +400 mg/L RAPA treatment.Results The A490 value of SRA01/04 was 0.680±0.020,0.550±0.013,0.480±0.014,0.400±0.011 and 0.200±0.019 in the control group,TGF-β2 group,TGF-β2 + 10 mg/LRAPA group,TGF-β2 + 100 mg/L RAPA group,TGF-β2 + 1000 mg/L RAPA and TGF-β2 + 10 000 mg/L RAPA group,respectively,showing a gradually declining trend in SRA01/04 rate of proliferation with increasing RAPA concentrations (F =101.920,P =0.000).RT-PCR and Western blot assay showed that the relative expression levels of α-SMA mRNA (α-SMA mRNA/β-actin mRNA)and protein (α-SMA/β-actin)in the cells were significantly increased in the TGF-β2 treatment group.However,with exposure to RAPA,the relative expression levels of α-SMA mRNA and protein were significantly lowered with increasing RAPA concentrations,but the expression levels of E-cad mRNA and protein were raised (α-SMA mRNA:F =294.660,P =0.000 ; α-SMA protein:F =346.950,P =0.000 ; E-cad mRNA:F =264.250,P =0.000 ; E-cad protein:F =317.327,P =0.000).In addition,after exposure to 400 mg/L RAPA,the expression levels of α-SMA protein gradually reduced and those of E-cad protein gradually increased with increasing treatment durations,showing significant differences among the different time points (α-SMA:F =693.864,P =0.000 ;E-cad:F=369.286,P =0.000).Conclusions RAPA can inhibit the proliferation of SRA01/04 in vitro and arrest EMT of SRA01/04 induced by TGF-β2 in a dose-and time-dependent manner.

Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

OBJECTIVE: To perform the validation and analysis of forensic parameters of Goldeneye DNA ID 26Y system. METHODS: Based on the validation rules of Scientific Working Group on DNA Analysis Methods (SWGDAM), the kit was assessed from several parts, as test of PCR system, reproducibility, accuracy, and sensitivity, etc. And Y-STR loci of 517 unrelated healthy individuals from Eastern China were genotypes by this kit. The distribution and frequency of haplotype were calculated and forensic parameters of the kit were assessed. RESULTS: The complete profiles can be obtained even when the PCR reaction volume with 6.25 microL. And correct profile was obtained with DNA down to 125 pg. No reproducible peaks were detected with the DNA of common animals and microorganism with the kit. For the male-male mixture testing, average 70% of the minor alleles were obtained when the ratios of 1:19 and 19:1. For the male-female mixture testing, results showed that the sensitivity of the kit was no compromised with the addition of female samples. CONCLUSION The validation studies demonstrated that Goldeneye DNA ID 26Y system has good sensitivity and specificity, and suitable for mixture testing. The polymorphism of 26 Y-STR loci included in this kit are good for forensic application.
Alleles ; Animals ; Asian People/genetics* ; China ; Chromosomes, Human, Y ; DNA ; DNA Fingerprinting/standards* ; Female ; Forensic Genetics/methods* ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVE: To investigate the genetic data of 21 autosomal STR included in Goldeneye™ DNA ID 22NC Kit in Chinese Han nationality and to evaluate the forensic application. METHODS: By detected 500 unrelated healthy individuals in Chinese Han nationality of East China with Goldeneye™ DNA ID 22NC Kit, allele frequencies, population genetics parameters and linkage disequilibrium information of the 21 autosomal STR were statistically analyzed. RESULTS: In the 21 autosomal STR, no deviations from Hardy-Weinberg equilibrium were detected and all loci were independent form each other. DP values of 21 autosomal STR were all above 0.85, and the combined discrimination power was 1-3.616 5 x 10(-26). Combined mean exclusion chance of this system in duo cases was 1-2.786 81 x10(-6), in trio cases was 1-8.545 82 x 10(-1). CONCLUSION Twenty-one autosomal STR included in Goldeneye™ DNA ID 22NC Kit are highly polymorphic in the Han nationality. Combined with Goldeneye™ DNA ID 20A Kit, the kit can satisfy the needs for full-sibling testing and facilitate the solution of this kind of case tools.
Alleles ; Asian People/genetics* ; China ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genetic Markers/genetics* ; Genetics, Population ; Genotype ; Humans ; Polymorphism, Genetic ; Reagent Kits, Diagnostic

To investigate the anticancer effects of ring C in 18β-glycyrrhetinic acid (GA), a series of GA derivatives featured with 9(11)-ene moiety in ring C were designed and synthesized. The structures were confirmed by IR, LC-MS and 1H NMR. Their inhibitory effects towards human prostate cancer PC-3 and leukemia HL-60 cell lines were determined. Most of the derivatives displayed stronger antiproliferative activities than GA. Particularly, compound 14 showed promising anticancer activity with the GI50 values of 4.48 µmol · L(-1) and 1.2 µmol · L(-1) against PC-3 and HL-60 cells respectively, which is worth further study.
Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Drug Design ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; HL-60 Cells ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; pathology

To study the link between BRAFV600E status and the expression of BIM gene in papillary thyroid carcinoma( PTC) tissues and to analyze the association of these factors with clinicopathological characteristics. BRAFV600E status was determined by MASA-PCR, and qPCR was applied to detect the expression of BIM gene. Finally, the associations of these factors with clinicopathological characteristics were analysed. The rate of mutant BRAFV600E in PTC was 54. 1% , and the expression of BIM gene was lowered in BRAFV600E positive PTC tissues. Additionally, there was significant association( P < 0. 05) between BRAFV600E positiveness and raised TNM Staging (Ⅲ/ Ⅳ), and lowered BIM expression was significantly associated (P<0. 05) with the tumor size and raised TNM Staging(Ⅲ/ Ⅳ). These findings may help us to know more about the mechanism of PTC and to develop new diagnostic biomarkers or prognostic indicators of PTC.

Objective To compare the clinical and pathological features of Chinese young breast cancer(age ≤ 35)with elder patients(> 35)using Meta analysis .Methods Published studies concerning clinical and pathological features of young breast cancer in China were searched systemically and assessed .Stata12 .0 software was used for data analyzing and calculating OR and its 95%CI .Results Totally 31 studies were selected for Meta analysis ,and most of them were classified as 6 - 7 scores ,which showed the quality of articles was high .The risk factors of breast cancer and its pooled odds ratio values with statistical significance were as fol‐lows 6 .42(95% CI :4 .22 - 9 .79) ,0 .61(95% CI :0 .50 - 0 .74)when clinical staging of 0 - Ⅱ phase or Ⅰ - Ⅱ phase ,2 .25(95% CI :1 .69 - 2 .99)when histological type of Invasive carcinoma ,1 .73(95% CI :1 .23 - 2 .43)when histological grade of III grade ,1 .80 (95% CI :1 .23 - 2 .43)when positive of lymph node metastasis .Conclusion Compared with elder breast cancer ,the clinical and pathological characteristics of young breast cancer were mainly for the high misdiagnosis rate ,the late clinical stage ,the high pro‐portion invasive carcinoma ,the poor histological differentiation and the lymph node metastasising easily ,the hint of young breast cancer screening and treatment may be different principles and measures should be adopted .

Objective To construct a lentiviral vector for RNA interference(RNAi)of MACC1 gene and to detect the best trans-fection condition by transfected into MB-231 cells .Methods The siRNA was designed and converted into cDNA of shRNA (small hair pin RNA) of siRNA for MACC1 gene .The cDNA was synthesized and inserted into pMAGic lentiviral plasmid vector which was linearized by enzyme Age Ⅰ and EcoRⅠ .The recombinant plasmid was transformed into competent E .coli DH5α cells .The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing .The recombinant lentivirus was packaged into mature lentivirus by 293FT cells and used to infect MB-231 cells .To detected the transfection condition of high efficiency of infection and low multiplicity of infection .Results PCR and sequencing verified that the recombinant lentivirus plasmid MACC1-shRNA was successfully constructed .The best transfection condition was MOI=40 by transfected into MB-231 .Conclu-sion The lentiviral RNAi expression vector targeting MACC 1 gene is successfully constructed and it can infect MB-231 cells effi-ciently ,which lays the experimental foundation for the research on the changes of malignant biological activity of tumor cell lines and gene therapy .

Objective To optimize the short tandem repeats(STR)which link closely to survival motor neuron(SMN)and have redundant polymorphism information contents,and to use these STR in the prenatal diagnosis of spinal muscular atrophy(SMA).Methods Eleven STR loci(D5S435,D5F153, DSF151,D5S637,D5S1413,D5S125,D5S464,D5S1556,DSF149,D5S351,MAP1B-5')were amplified by PCR.Then the PCR products were detected by polyacrylamide gel electrophoresis(PAGE)and analyzed by silver staining.STR loci were evaluated and optimized by their PIC values.PCR-PAGE and gene scan were combined to make genetic link analysis for SMA families based on the optimized STR.Results Three STR loci(D5S435,DSF149 and D5S351)were selected with 8,19 and 18 polymorphic fragments detected respectively in 100 normal individuals.Their PIC values were 0.84,0.91 and 0.92 respectively.Four carriers and 2 normal individuals were detected from 6 SMA families with linkage analysis by using the 3 STR.Conclusion This genetic diagnosis system based on the 3 STR loci can provide rapid prenatal diagnosis for SMA families,can eliminate maternal blood contamination,and also can discriminate carriers from normal individuals in the fetuses,which makes the prenatal diagnosis system of SMA perfect.