Mechanism of ELL-associated factor 2 and vasohibin 1 regulating invasion, migration, and angiogenesis in colorectal cancer.

BACKGROUND: As a novel endogenous anti-angiogenic molecule, vasohibin 1 (VASH1) is not only expressed in tumor stroma, but also in tumor tissue. Moreover, studies have shown that VASH1 may be a prognostic marker in colorectal cancer (CRC). Knockdown of VASH1 enhanced transforming growth factor-ß1 (TGF-ß1)/Smad3 pathway activity and type I/III collagen production. Our previous findings suggest that ELL-associated factor 2 (EAF2) may play a tumor suppressor and protective role in the development and progression of CRC by regulating signal transducer and activator of transcription 3 (STAT3)/TGF-ß1 signaling pathway. However, the functional role and mechanism of VASH1-mediated TGF-ß1 related pathway in CRC has not been elucidated. AIM: To investigate the expression of VASH1 in CRC and its correlation with the expression of EAF2. Furthermore, we studied the functional role and mechanism of VASH1 involved in the regulation and protection of EAF2 in CRC cells in vitro. METHODS: We collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein and VASH1 protein in patients with advanced CRC. Following, we investigated the effect and mechanism of EAF2 and VASH1 on the invasion, migration and angiogenesis of CRC cells in vitro using plasmid transfection. RESULTS: Our findings indicated that EAF2 was down-regulated and VASH1 was up-regulated in advanced CRC tissue compared to normal colorectal tissue. Kaplan-Meier survival analysis showed that the higher EAF2 Level group and the lower VASH1 Level group had a higher survival rate. Overexpression of EAF2 might inhibit the activity of STAT3/TGF-ß1 pathway by up-regulating the expression of VASH1, and then weaken the invasion, migration and angiogenesis of CRC cells. CONCLUSION: This study suggests that EAF2 and VASH1 may serve as new diagnostic and prognostic markers for CRC, and provide a clinical basis for exploring new biomarkers for CRC. This study complements the mechanism of EAF2 in CRC cells, enriches the role and mechanism of CRC cell-derived VASH1, and provides a new possible subtype of CRC as a therapeutic target of STAT3/TGF-ß1 pathway.

Overexpression of ELL-associated factor 2 suppresses invasion, migration, and angiogenesis in colorectal cancer.

BACKGROUND: The androgen responsive gene, ELL-associated factor 2 (EAF2), expressed in benign prostate tissues, has been shown to play an important role in tumor suppression in a variety of malignant tumors. In addition, some scholars found that EAF2 frameshift mutations are associated with intratumor heterogeneity in colorectal cancer (CRC) and inactivation of EAF2 in microsatellite instability-high CRC. However, the molecular mechanism by which EAF2 is involved in CRC invasion and metastasis remains unclear. AIM: To determine the clinical value of expression of EAF2 protein in CRC, and to study the effects of EAF2 on the invasion, migration, and angiogenesis of CRC cells in vitro. METHODS: In this study, we collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein in patients with advanced CRC. Subsequently, we investigated the effect of EAF2 on the invasion, migration, and angiogenesis of CRC cells in vitro using plasmid transfection. RESULTS: EAF2 protein was lowly expressed in cancer tissues of patients with advanced CRC. Kaplan-Meier survival analysis showed that the survival rate of the high EAF2 level group was higher than that of the low EAF2 level group. CONCLUSION: Our results demonstrated that EAF2, as a tumor suppressor, may inhibit the invasion, metastasis, and angiogenesis of CRC cells by regulating the signal transducer and activator of transcription 3/transforming growth factor-ß1 crosstalk pathway, and play a cancer suppressive and protective role in the occurrence and development of CRC. Our findings are of great significance to provide a new idea and theoretical basis for the targeted diagnosis and treatment of CRC.

Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer.

BACKGROUND: Drugs targeting mitochondria can induce mitophagy and restrain proliferation in colorectal cancer (CRC) cells. Phosphoglycerate mutase family member 5 (PGAM5) activates serine/threonine PTEN-induced putative kinase 1/Parkin pathway-mediated mitophagy. However, there are few studies on the clinical and prognostic significance of expression of PGAM5 protein and mitophagy-related protein Parkin in patients. AIM: To assess the clinical significance of PGAM5 and Parkin proteins, as biomarkers for diagnosis and prognosis of CRC, by studying their expression in advanced CRC tissues and their association with clinicopathological parameters. METHODS: The expression of PGAM5 and Parkin in CRC tissues from 100 patients was determined by immunohistochemistry. Each case was evaluated by using a combined scoring method based on signal intensity staining (scored 0-3) and the proportion of positively stained cancer cells (scored 0-4). The final staining score was calculated as the intensity score multiplied by the proportion score. Specimens were categorized as either high or low expression according to the Youden index, and the association between the expression of PGAM5 or Parkin and clinicopathological factors was ascertained. Additionally, we employed western blot to measure PGAM5 and Parkin protein expression in six matched pairs of CRC and adjacent non-tumor tissues. RESULTS: Immunohistochemical and western blot findings showed that both PGAM5 and Parkin protein expression in tumor tissues was significantly higher than that in the adjacent tissues: PGAM5 and Parkin were mainly expressed in the cytoplasm of colonic epithelial cells. PGAM5 and Parkin protein levels were significantly positively correlated in advanced CRC tissues. Moreover, reduced Parkin protein expression was an independent prognostic factor for overall survival and progression-free survival in CRC patients as evinced by multivariate analysis. CONCLUSION: The expression of PGAM5 protein and mitophagy-related protein Parkin has diagnostic significance for CRC and may become new biomarkers. Parkin may be a potential marker for the survival of CRC patients.

Review of research progresses in genetics related to myopia / 国际眼科杂志(Guoji Yanke Zazhi)

@# Myopia is an extremely common state of refractive error, and the incidence of myopia is increasing year by year and the age of onset is usually earlier. It has grown up to be a major public health problem that endangers people's visual health. Studies have demonstrated that myopia is a multi-factor complex disease which affected by genetic factors, environmental factors and gene-environment interaction. Genetics includes classical genetics and epigenetics. The emergence of epigenetic research has opened a new perspective of basic research on myopia. In recent years, the researchers have proposed that the occurrence of myopia may be related to epigenetics, and more and more experimental studies have also proved this view. The genetic study of myopia has identified several myopia genes and candidate genes for high myopia and myopia by using linkage and genome-wide association methods, which greatly deepened the understanding of the genetic basis of myopia. This article reviews the research on classical genetics and epigenetics of myopia in recent years.

[Distribution of tap1 and tap2 loci allelic and genotypic frequencies in Chinese Hans].

This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on.

[Estimating genetic distance and phylogenetic tree of HPA-1-3, 5, and 15 in different populations].

According to the human platelet alloantigens (HPA) polymorphisms in five systems, the distributions of HPA-1 -3, 5, and 15 systems in 1 000 Chinese donors were carried out by using a polymerase chain reaction with sequence-specific primers (PCR-SSP) method. The genetic distance and phylogenetic tree between Chinese Hans and other populations were estimated by using DISPAN and PHYLIP software. As presented by the phylogenetic tree, Asian had a convergence with European first, and grouped together with African. Beninese which came from Africa was on the top of dendrogram. Indian was located between Asian and European. Brazilian was converged with other Europe populations. Oceanian Polynexiya had been shown specifically to cluster with Asia populations. These results proved the "out of Africa theory" from one side, and it also confirmed that early migration of Asian is from south to southeast, and east Asia., thus it is probable that Europeans are migrated from south to north, and west Europe. As genetic distance was estimated effectively by HPA systems, HPA systems could serve as the genetic marker in human migration and evolution research.

[Polymorphism of the human platelet alloantigens HPA-3 and HPA-9w in the Chinese Han population].

OBJECTIVE: To study the polymorphism of human platelet alloantigens HPA-3 and HPA-9w in the Chinese Han population. METHODS: A total of 1000 unrelated Chinese Han blood donors from different provinces of China were genotyped for HPA-3 and HPA-9w using PCR-sequence specific primer assay. RESULTS: Gene frequencies of 1000 Chinese Hans for HPA-3a and HPA-3b were 0.5935 and 0.4065 respectively, and all of them were HPA-9a positive. The distributions of HPA-3, HPA-9w of Chinese Hans which detected by chi-square criterion fit Hardy-Weinberg equilibrium. There were significant differences of the HPA-3 alleles gene frequency between Guangdong province and other five investigated provinces which included Shanxi, Heilongjiang, Zhejiang, Yunnan and Jiangsu. In comparison to other ethnic groups, no significant differences were observed in the distributions of HPA-3 except the Vietnamese and Australian. CONCLUSION: The results show that the chance of HPA-3 incompatibility were 0.3661 in random transfusion, and also provide a basis for researching on alloimmune thrombocytopenia and HPA-matched transfusion.

[Case report of a rare platelet-specific antigen HPA-10bw allele found in Chinese mainland].

A total of 1,000 Chinese blood donors were typed for human platelet antigens (HPA) using a sequence specific primers -polymerase chain reaction (PCR-SSP) based HPA genotyping method. An individual with a rare HPA-10w(a+b+) genotype was found. In order to confirm the typing results, a fragment of HPA-10 gene was amplified by PCR and then sequenced. Sequencing data showed that a single G to A substitution at nucleotide 263 occurred, resulting in amino acid change from Arg(CGA) to Gln(CAA) at position 62 of GPa protein. The substitution generated antigenic specificity HPA-10bw. The detection of an HPA-10bw allele in the Chinese population suggests that this rare allele should be considered in platelet alloimmunization, such as neonatal alloimmune thrombocytopenia (NAIT), post-transfusion thrombocytopenic purpura (PTP) and post-transfusion refractoriness to platelets (PTR).

[Advances in the studies on human platelet alloantigen--review].

Human platelet alloantigens (HPA) are specific antigens carried by platelet glycoproteins, which genes showing single nucleotide polymorphism. HPA can induce alloantibodies bringing about alloimmune response. They play important roles in post-transfusion refractoriness to platelets, post-transfusion thrombocytopenic purpura, fetomaternal alloimmune thrombocytopenia, and graft-versus-host disease. Because of their side effects in clinical blood-transfusion, there were a great deal of studies on HPA during last few decades. This review focuses on the nomenclature of HPA, the polymorphisms of platelet glycoproteins, HPA typing of the serological and molecular technology, as well as the mechanism of alloimmunization to HPA and correlated diseases.

Curative effects of interferon-alpha and HLA-DRB1 -DQA1 and -DQB1 alleles in chronic viral hepatitis B.

AIM: To investigate the association between curative effects of interferon-alpha and partial human leucocyte antigen (HLA) II alleles in chronic viral hepatitis B. METHODS: Sixty patients with chronic viral hepatitis B in Shanghai were treated with a standard course of treatment with interferon-alpha for 6 mo. HLA-DRB1, -DQA1, and -DQB1 alleles were detected by polymerase chain reaction-sequence specific primer (PCR-SSP) method. RESULTS: Frequencies of HLA-DRB1*04 (P<0.025) and HLA-DQA1*0303 (P<0.01) in non-responders were significantly higher than those in partial and complete responders. Frequencies of HLA-DQA1*0505 (P<0.025) and HLA-DQB1*0301 (P<0.005) in partial and complete responders were significantly higher than those in non-responders. CONCLUSION: Non-response to interferon-alpha therapy is positively correlated with HLA-DRB1*04 and HLA-DQA1*0303, and negatively correlated with HLA-DQA1*0505 and -DQB1*0301 in patient with chronic viral hepatitis B. HLA II genes of the identification alleles provide a method for evaluating outcome of interferon-alpha treatment.

[The polymorphism distributions of MICA and MICB microsatellite in Guangdong Han population].

OBJECTIVE: To investigate the genetic polymorphism of microsatellite in the exon 5 of MICA gene and the intron 1 of MICB gene in Guangdong Han population. METHODS: One hundred and six samples of Guangdong Han population were genotyped by polymerase chain reaction and fluorescent technique (6-FAM). Gene frequency, power of discrimination, expected heterozygosity, polymorphism information content and probability of paternity exclusion were calculated. RESULTS: The genotype distributions of MICA and MICB microsatellite met Hardy-Weinberg equilibrium. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). The genotype distribution frequencies of A5-5.1 (14.15%) and A5-5 (10.38%) are high. MICB CA14 was the most common allele (0.3255), and CA19,28 was the least popular one (0.0047). CA27 was not observed. The genotype distribution frequency of CA14-CA14(14.15%) is high. CONCLUSION: The microsatellite of the exon 5 of MICA gene and the intron 1 of MICB gene could be used as the genetic markers of Chinese population in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

[The study on the haplotype of MICA and MICB microsatellite locus in Guangzhou Han population].

This study is to investigate genetic polymorphisms and haplotypes of microsatellite locus in the exon 5 of the MICA gene and intron 1 of the MICB gene based on 106 samples of Guangzhou Han Population by polymerase chain reaction and fluorescent technique (6-FAM). The corresponding haplotype frequencies, linkage disequilibria values and relative linkage disequilibria values were estimated based on population data. The results show that the genotype distributions of MICA and MICB microsatellite meet Hardy-Weinberg equilibrium in Guangdong Han population. In total, 5 alleles of MICA microsatellite locus and 14 alleles of MICB microsatellite locus were observed. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). MICB CA14 was the most common allele (0.3255), and CA19 and CA28 were the least popular ones (0.0047). CA27 was not observed. Twenty-one kinds of MICA-MICB haplotypes occurred at frequencies of more than 1% (linkage disequilibria value>0). The common MICA-MICB haplotypes were A5-CA14 (16.73%) , A5.1- CA18 (8.75%), A4- CA26(3.76%),A9-CA15(3.66%) and A6-CA21(2.61%) (chi(2)>3.84, P<0.05) , and they were strong linkage disequilibria. The polymorphisms and haplotypes distributions of MICA and MICB microsatellite locus in Guangzhou Han population have their own genetic characteristics. The microsatellite locus of the exon5 of the MICA gene and intron 1 of the MICB gene could be used as the genetic markers in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

[Study on HLA haplotypes in Jiangsu-Zhejiang-Shanghai Han population].

This study is to investigate HLA haplotypes in Jiangsu-Zhejiang-Shanghai Han population based on 166 families by serological and molecular biological HLA typing methods and to analyze the distribution characteristic of HLA haplotypes. The results showed that allele frequencies of more than 10% for HLA antigens were A2, A11, A24, B13, B46, B60, DRB1 *04, DRB1 *08, DRB1* 09, DRB1 * 12 and DRB1 * 15. In the analysis of HLA haplotypes, 128 kinds of A-B haplotypes and 182 kinds of B-DRB1 haplotypes were found, comprising 19.28% (128/664) and 27.41%(182/664) of total theoretical haplotypes, respectively. 18 kinds of A-B haplotypes and 23 kinds of B-DRB1 haplotypes occurred at frequencies of more than 0.5% (linkage disequilibrium value, delta > 0) .351 kinds of A-B-DRB1 haplotypes were found, comprising 52.86% (351/664) of total theoretical haplotypes, and 8 kinds of A-B-DRB1 haplotypes occurred at frequencies of more than 0.5% (delta > 0). The common A-B-DRB1 haplotypes were A30-B13-DRB1 * 07(4.22%), A2-B46-DRB1 * 09(3.77%), A33-B58-DRB1 * 17(3.01%), A33-B58-DRB1 * 13.1 (1.81%) and A11-B75-DRB1 * 12(1.51%). The HLA haplotype distribution of Jiangsu-Zhejiang-Shanghai Han population has its own genetic characteristics, so it suggests this population is between southern and northern Han population. The HLA polymorphism of Chinese Han population is more abundant in East Asian populations.

[The genetic characteristic of HLA-DRB1 locus in the Jiangsu-Zhejiang-Shanghai Han population and a comparison of its frequency distribution with that of other populations].

OBJECTIVE: To investigate the genetic polymorphism of HLA-DRB1 locus in Jiangsu-Zhejiang-Shanghai Han population and analyze the characteristic of the allele frequency distribution in comparison with that of other populations. METHODS: The technique of polymerase chain reaction-sequence specific primers (PCR-SSP) and reverse polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) was adopted in genotyping a sample of 626 unrelated healthy individuals collected from a Chinese Han population in Jiangsu-Zhejiang-Shanghai area. HLA-DRB1*0101-1001, DRB3, DRB4 and DRB5 were detected. The allele frequency of HLA-DRB1 was calculated, and the allele frequency distribution of HLA-DRB1 in this population was compared with the results from other populations. RESULTS: HLA-DRB1*0101, 0301, 0701, 09012, 1001, 1201, 1202, 1301/02, 1303/04, 1401/04/05, 1402/03/1305, 1501/02, 16021 and 04xx, 08xx were detected in Jiangsu-Zhejiang-Shanghai Han population. The common HLA-DRB1*allele included 09012(17.97%), 04xx(12.53%), 1202(11.42%) and 1501/02(11.02%). The polymorphism information content is 0.9024, and expected heterozygosity is 0.9634 in Jiangsu-Zhejiang-Shanghai Han population. CONCLUSION: The HLA-DRB1 distribution of Jiangsu-Zhejiang-Shanghai Han population shares some genetic characteristic with other Han populations, but it exhibits its own characteristic, suggesting the intermediate state of this population between the southern and northern Han populations. The polymorphism of HLA-DRB1 of Jiangsu-Zhejiang-Shanghai Han population is the most abundant one in this study.

[Activation of random-donor platelet concentrates and platelet prepared from random-donor apheresis collections during storage].

The activation of random-donor platelet concentrates and platelets prepared from random-donor apheresis collections (plateletpheresis) during storage were studied. Percentage of CD62p staining and mean channel fluorescence (MCF) of CD41 of two kinds of platelets during storage on day 0, day 1, day 3 and day 5 were determined by flow cytometry. The results showed that percentages of CD62p staining and MCF of CD41 in plateletpheresis were (18.91 +/- 6.25)%, (19.48 +/- 8.27)%, (22.82 +/- 6.06)%, (56.71 +/- 11.79)% and (8.09 +/- 2.38)%, (8.13 +/- 2.45)%, (8.44 +/- 2.51)%, (19.87 +/- 6.13)%, while the results of platelet concentrates were (30.65 +/- 12.33)%, (31.46 +/- 11.86)%, (32.51 +/- 13.05)%, (63.55 +/- 13.27)% and (10.33 +/- 4.37)%, (11.09 +/- 6.61)%, (13.46 +/- 9.69)%, (24.41 +/- 10.15)%, respectively. The platelet count and pH value were also determined. The platelet number, pH value, percentage of CD62p staining and MCF of CD41 had no significant difference within 3 days of platelet storage. The platelet number and pH value decreased significantly (P < 0.001), while percentages of CD62p staining and MCF of CD41 increased significantly (P < 0.001) on day 5 of storage. It is concluded that the quality of plateletpheresis is better than platelet concentrate.

[The polymorphism distributions of nine STR loci in Shanghai Han population].

By multiplex amplification and four fluorescent technique,the polymorphism distributions of nine STR loci, D3S1358, vWA,FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820 were investigated in Shanghai Han population.Gene frequency (Pi),power of discrimination (DP),polymorphism information content (PIC) expected heterozygosity (H) and probability of paternity exclusion (PE) were calculated. All loci meet Hardy-Weinberg equilibrium. DP of FGA locus,H of D8S1179 locus,PIC of D18S51 locus and PE of D18S51 locus are the biggest among nine STR loci. Cumulate DP (CDP) of nine STR loci is 0.9999996, Cumulate PE (CPE) of nine STR loci is 0.99991. Nine STR loci could be used as the genetic markers of Chinese population in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

[Typing of HLA-AB by Reverse PCR-SSOP and Clinical Application]

A rapid and accurate method of DNA typing for HLA was established to compensate the unsatisfactory serological typing for HLA before transplantation. DNA typing for HLA using by reverse polymerase chain reaction with sequence-spcific oligo probe (reverse PCR-SSOP) could detect HLA-A(*0101 - 8001) and B(*07021 - 8201). The results showed that HLA-AB alleles were successfully analysed in 60 matching subjects and 16 control DNAs from UCLA by reverse PCR-SSOP without false negtive and false positive results. The results were concordance with those of UCLA. The error rate of serological typing was 6.4% for HLA-A and 7.4% for HLA-B. The serological typing missed HLA-A24 and HLA-B46 for two patients with leukemia respectively. Our results suggest that DNA typing for HLA by reverse PCR-SSOP has proved to be a high-resolution, high-specificity, rapid and accurate technique, suitable for clinical application with a greater precision than serological typing.