Objective:To explore the effect of siRNA targeting myeloid cell leukemia-1(Mcl-1)on the biological behavior of salivary adenoid cystic carcinoma cells.Methods:The chemically synthesized Mcl-1-siRNA was transfected into salivary adenoid cystic carci-noma SACC-2 cells.The expression levels of Mcl-1-mRNA and Mcl-1protein were examined by Real-time PCR and western blotting respectively.MTT assay,transwell chamber and flow cytometry were used to determine the effect of Mcl-1-siRNA on SACC-2 cell pro-liferation,migration and apoptosis.Results:Compared with the control group,liposome group and NC-siRNA group,SACC-2 cell proliferation rate of Mcl-1-siRNA group was obviously slowed down.48 h after transfection,the migration of SACC-2 cells in Mcl-1-siRNA group(39 ±9.0)were lower than that in control group(69 ±6.0).The apoptosis rate of Mcl-1-siRNA group(8.6%)was sig-nificantly higher than that in control group(1.9%).Conclusion:Silence Mcl-1 can inhibit cell proliferation and migration and pro-mote apoptosis of salivary adenoid cystic carcinoma cells.

Objective:To explore the effects of cysteine-rich 6 1 (Cyr6 1 )on biological behavior of human adenoid cystic carcinoma ACC-LM and ACC2 cells.Methods:The chemically synthesized Cyr6 1-siRNA was transfected into ACC-LM and ACC2 cells.Cell proliferation was measured by the MTT method,the invasive ability was evaluated by Transwell chamber assay,and cell apoptosis was analyzed using flow cytometry by double staining with Annexin V and propidium iodide.Results:Cyr61-siRNA significantly down-regu-lated Cyr61 protein expression in ACC-LMand ACC2 cells.Cyr61-siRNA markedly inhibited the proliferation and invasion of the cells, however,there was no significant difference in cell apoptosis between Cyr6 1-siRNA and control groups.Conclusion:Cyr6 1 promote the proliferation and invasion of adenoid cystic cancer cells.

Objective:To investigate the effect of c-fos on the proliferation and migration of oral squamous cell carcinoma and potential mechansism.Methods:The expression of c-fos,CyclinD1 and p16 in 60 oral squamous cell carcinoma samples and 60 oral mucosa tissue samples was examined by immunohistochemistry.HN6 and SCC9 cells were respectively transfected with siRNA-c-fos and siRNA-scramble,then were respectively divided into control group,siRNA-scramble group and siRNA-c-fos group.The mRNA and protein expressions of CyclinD1 and p16 were decteted,meanwhile cell proliferation and migration were tested.Results:Compared with the oral mucosa tissue samples,the expressions of CyclinD1 and c-fos were increased in the carcinoma samples,while the expression of p16 was reduced.Compared with control group,the expressions of CyclinD1 in siRNA-c-fos group were significantly reduced,while p16 enpression was increased,with the inhibition of cell proliferation and migration.Conclusion:c-fos may regulate pl6/CyclinD1 signaling pathways and promote the proliferation and migration of oral squamous cell carcinoma.

Objective:To investigate the effects of IGF-1 and IL-1β on the proliferation and apoptosis of cultured human condylar chondrocytes( CCs) of temporomandibular joint( TMJ) . Methods:Cultured CCs were derived from human TMJ condylar cartilage tis-sue, and identified by immunocytochemistry staining. The cultured cells were divided into 6 groups:control group, IL-1β(10 μg/L) group and IL-1 group (10 μg/L) + IGF-1 group (0, 1, 10, 50 and 100 μg/L, respectively). The cell proliferation ability was de-tected by MTT assay. The cell cycle and apoptosis were detected by flow cytometry. The expression of apoptosis-associated factors Bcl-2, Bax and p38 MAPK/NF-κB proteins were detected by Western blot. Results:Type II collagen was positively expressed in cultured CCs. IL-1β treatment decreased cell proliferation, increased cell apoptosis with concomitant increase of the percentage of early apopto-sis and late apoptotic cells, increased Caspase-3 expression, decreased Bcl-2/Bax ratio, and increased the expression of p38 MAPK/NF-κB proteins. Whereas, with 1-100 μg/L IGF-1 pretreatment, the proliferation ability and Bcl-2/Bax ratio of the cells were in-creased(P<0. 05), the apoptotic cells were decreased, the expression of Caspase-3 and p38 MAPK/NF-κB proteins was decreased ( P<0. 05) in a dose-dependent manner. Conclusion:IGF-1 may inhibit IL-1β-induced cell apoptosis and attenuate the activation of p38 MAPK/NF-κB of human condylar chondrocytes.

OBJECTIVETo detect potential mutations of fibrillin-1 (FBN1) gene in a child with Marfan syndrome (MFS) and explore its molecular pathogenesis.

METHODSThe 66 exons of the FBN1 gene were analyzed by direct sequencing. SIFT and PolyPhen-2 were used to predict the structural and functional changes at the protein level.

RESULTSA novel heterozygous mutation c.3998 G>A (p.Cys1333Tyr) was found in exon 32 in the child. The same mutation was not found among his unaffected family members and 683 healthy controls. Multiple sequence alignment showed that this novel mutation was located in a highly conserved region of the FBN1 protein across various species and may induce structural change to a functional domain.

CONCLUSIONThe novel c.3998G>A (p.Cys1333Tyr) mutation of the FBN1 gene probably predisposed the MFS in the child. Above finding has enriched the spectrum of FBN1 mutations.

OBJECTIVE: To screen for MYOC gene variants among sporadic patients with primary open angle glaucoma (POAG). METHODS: For 398 patients with POAG, Sanger sequencing was applied to detect potential variants of the MYOC gene. RESULTS: Eight patients (2.0%) were found to harbor variations of the MYOC gene. These included five types of variants, among which c.667C>T (p.Pro223Ser) and c.1138G>T (p.Asp380Tyr) were novel. c.382C>T (p.Arg128Trp), c.1109C>T(p.Pro370Leu) and c.1130C>A (p.Thr377Lys) were previously associated with POAG. Alignment of amino acid sequences of MYOC proteins of various species revealed that the two novel variants have occurred at highly conserved positions. c.1138G>T was predicted to be possible pathogenic by Bioinformatic analysis. CONCLUSION Two novel variants of the MYOC gene were detected among sporadic POAG patients, which enriched its variant spectrum.
Cytoskeletal Proteins ; genetics ; Eye Proteins ; genetics ; Glaucoma, Open-Angle ; genetics ; Glycoproteins ; genetics ; Humans ; Mutation

Objective To explore the rare nonsynonymous variants of ABCA1 gene in primary open angle glaucoma (POAG).Methods A prospective cohort study was carried out.Three hundred and ninety-eight POAG patients and 198 healthy controls matched in age and gender were recruited from March 2017 to March 2018 in Eye and Ear Nose Throat (ENT) Hospital of Fudan University.The periphery blood of 2-5 ml from all the subjects was collected for extraction of DNA,and rare variant analysis of the ABCA1 gene was conducted by whole exome sequencing (WES) data of these subjects.The study protocol was approved by Ethic Committee of Eye and Ear Nose Throat Hospital of Fudan University and Sichuan Provincial People's Hospital (No.2016-32-1,and written informed consent was obtained from each subject prior to entering the study cohort.Results A total of 21 rare nonsynonymous variants (minor allele frequency MAF＜0.O1) were detected in the coding regions of ABCA1 gene in 27 subjects of the 398 POAG,with the detection rate of 6.8％.Among them,c.4310C＞A (p.Thr1437Asn),c.3772G＞T(p.Asp1258Tyr),c.775A＞G (p.Lys259Glu) and c.1507_1508insGAGGT (p.Glu503GlyfsX7) were four novel variants.In the 198 healthy controls,five rare nonsynonymous variants were detected in the ABCA1 gene from five subjects respectively,with the detection rate of 2.5％,the detection rate of nonsynonymous in POAG group was higher than that in healthy control group,showing a significant difference (x2=4.72,P =0.03,OR =2.81).Conclusions Rare nonsynonymous variants in ABCA1 is associated with the pathogenesis of POAG.These variants can enrich the variation spectrum of ABCA1.

Objective:To detect whether Toll-like receptor 4 ( TLR4) polymorphisms contributed to primary open angle glaucoma (POAG) in a Chinese population. Methods:A Chinese cohort, including 799 unrelated POAG patients and 799 unrelated controls, was enrolled in our case-control association study. The data was collected at Sichuan Provincial People's Hospital from May 2014 to March 2018. TLR4 functional single nucleotide polymorphisms (SNPs), including rs4986790 and rs4986791, were genotyped by SNaPshot method. Genotype and allele frequencies of the two SNPs were evaluated. This study was approved by the Institutional Review Boards of the Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital (No.2016-58), and complied with the guidelines of the Declaration of Helsinki. Written informed consents were obtained from all subjects prior to the study. Results:Allelic association analysis revealed that there were no significant association detected in the allelic distributions between the POAG cases and controls for SNPs rs4986790 ( P=0.317) and rs4986791 ( OR=1.000, 95% CI =0.062 5-16.002 2, P=1.000) in the TLR4 gene. Conditional analysis of the two SNPs did not show any significant difference in genotype and allele frequency between the case and the control groups. No association of the two SNPs with POAG was detected under four different genetic models, including homozygote, heterozygote, dominant and recessive models. Conclusions:Polymorphisms rs4986790 and rs4986791 in the TLR4 gene are not related to POAG in the Chinese cohort.

Objective:To investigate the association between rs284489 in the 8q22 region and primary open angle glaucoma (POAG) in Sichuan, and the association between rs284489 and gender difference.Methods:A case control study was adopted.A total of 894 Han Nationality POAG patients in Sichuan People's Hospital from September 2015 to March 2017 were included, and 994 control patients who participated in physical examination in the same period were included.All subjects had no blood relationship and all were Han Chinese.Each sample of 4 ml-peripheral blood was collected for extracting DNA and rs284489 information was obtained from NCBI website.Primers 5.0 software was used to design primers.Genotyping was performed by using a tailored "Chinese-Chip" for association analysis of the rs284489 in the 8q22 region.Genotype allele frequencies and Hardy-Weinberg equilibrium (HWE) were assessed by using χ2 test.Logistic regression was applied to adjust for gender differences between the cases and controls.The PS; Power and Sample Size Calculation (version 3.1.2) software was used to calculate statistical power.This study followed the Declaration of Helsinki.This study followed the guidelines for the collection of human genetic disease specimens issued by the Ministry of Health of China.The study protocol was approved by the Ethics Committee of Sichuan Provincial People's Hospital (No.2016-58). Results:The allele distribution of rs284489 was within the HWE for both case and control groups (both at P>0.05). The difference of the minor allele-G distribution between the case group and the control group was not significant (allelic P*=0.94, OR [95% CI] **=1.01[0.83-1.23]); To further investigate the association between rs284489 and POAG, four genetic models, including model 1 (AG vs. AA), additive model 2 (GG vs.AA), dominant model (GG+ AG vs.AA), and recessive model (GG vs.AG+ AA) were applied.There was no significant difference in the four genetic models between the case and control groups (adjusted P# additive model 1 =0.26, P# additive model 2 =0.54, P# dominant model =0.50, P# recessive model =0.25); the gender difference in this study was not associated with the polymorphism of rs284489 (adjusted P#=1.00, crude OR [95% CI]=1.00[0.88-1.14], adjusted OR [95% CI]=1.00 [0.87-1.14]). Conclusions:rs284489 is not statistically associated with POAG in a Sichuan Han Chinese population.

OBJECTIVE: To assess the association of 7 single nucleotide polymorphisms (SNPs) including rs13278062 (TNFRSF10A), rs3750846 (ARMS2-HTRA1), rs429358 (APOE), rs5817082 (CEPT), rs2043085 (LIPC), rs1626340 (TGFBR1), and rs8135665 (SLC16A8) identified through genome-wide association study (GWAS) with age-related macular degeneration (AMD) among ethnic Han Chinese from Sichuan, China. METHODS: A cohort of 576 AMD patients and 572 healthy controls were enrolled in a case-control study. The SNPs were genotyped by a Mass array MALDI-TOF System. On the premise that the genotype distribution of each SNP locus in both groups satisfied Hardy-Weinberg equilibrium, the genetic pattern was analyzed and the scores of allele and genotype frequencies ware compared. RESULTS: There was a significant association between TNFRSF10A rs13278062 and AMD under the heterozygous model (P = 0.000, OR = 1.529, 95%CI = 1.196-1.954) and the dominant model (P = 0.002, OR = 1.459, 95%CI = 1.154-1.865), suggesting that subjects carrying rs13278062GT and rs13278062TT + GT are more likely to develop the AMD, whereas no significant difference was observed for rs13278062 under other models. No association was detected with the other six SNPs and AMD under various genetic models. CONCLUSION This case-control association study has indicated that TNFRSF10A rs13278062 is associated with AMD under the heterozygous and dominant models, suggesting that the TNFRSF10A variant may be involved in the development of AMD among ethnic Han Chinese population.
Case-Control Studies ; Gene Frequency ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Genotype ; High-Temperature Requirement A Serine Peptidase 1/genetics* ; Humans ; Macular Degeneration/genetics* ; Polymorphism, Single Nucleotide