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With the gradual liberalization of the three-child policy and the development of assisted reproductive technology in China, the number of women with high-risk pregnancies is gradually increasing. In this study, 4211 fetuses who underwent chromosomal microarray analysis (CMA) with high-risk prenatal indications were analysed. The results showed that the overall prenatal detection rate of CMA was 11.4% (480/4211), with detection rates of 5.82% (245/4211) for abnormal chromosome numbers and 5.58% (235/4211) for copy number variants. Additionally, the detection rates of clinically significant copy number variants were 3.78% (159/4211) and 1.8% (76/4211) for variants of uncertain significance. The detection rates of fetal chromosomal abnormalities were 6.42% (30/467) for pregnant women with advanced maternal age (AMA), 6.01% (50/832) for high-risk maternal serum screening (MSS) results, 39.09% (224/573) with abnormal non-invasive prenatal testing (NIPT) results, 9.21% (127/1379) with abnormal ultrasound results, and 5.1% (49/960) for other indications. Follow-up results were available for 4211 patients, including 3677 (3677/4211, 87.32%) whose infants were normal after birth, 462 (462/4211, 10.97%) who terminated their pregnancy, 51 (51/4211, 1.21%) whose infants were abnormal after birth, and 21 (21/4211, 0.50%) who refused follow-up. The results of this study demonstrate significant variation in the diagnostic rate of chromosomal microarray analysis across different indications, providing valuable guidance for clinicians to assess the applicability of CMA technology in prenatal diagnosis.

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Objective To investigate the mechanism underlying the immunosuppressive effect and its reverse of γδ1 T cells derived from breast cancer tissues by inducing immunosenescence. Methods After γδ1 T cells isolated from breast cancer tissues were co-cultured with peripheral blood-derived naive CD4+ T cells, the proliferation of treated CD4+ T cells was detected by CCK-8 assay, and the activity of senescence-associated ß-galactosidase (SA-ß-Gal) in treated CD4+ T cells was detected by SA-ß-Gal staining. After the induced senescent CD4+ T cells were co-cultured with the naive CD4+ T cells, the proliferation, apoptosis, activity of the treated CD4+ T cells were examined by CCK-8 assay, flow cytometry and SA-ß-Gal staining, respectively; the expression of cell cycle-associated proteins P53, P21 and P16 in the treated CD4+ T cells was detected by Western blot analysis in order to verify the immunosuppressive effect of the senescent CD4+ T cells. The expression levels of inhibitory cytokines interleukin 17D (IL-17D), IL-10, transforming growth factor-α (TGF-α) and so on, in the supernatant were analyzed by cytokine microarray after co-culture of γδ1 T cells with naive CD4+ T cells. After the co-culture system of γδ1 T cells and naive CD4+ T cells was treated with anti-IL-17D monoclonal antibody, the proliferation of the naive CD4+ T cells was determined by CCK-8 assay. The level of IL-17D secreted by γδ1 T cells treated by single-stranded RNA40 (ssRNA40), the ligand of Toll like receptor 8 (TLR8) and TLR8 short hairpin RNA (shRNA) lentiviral vector was detected by ELISA. The effects of ssRNA40 on the immunosenescence induced by γδ1T cells were confirmed by SA-ß-Gal staining and Western blot analysis. Results γδ1 T cells derived from breast cancer tissues induced the immunosenescence of the naive CD4+T cells, and the senescent CD4+ T cells further exerted the immunosuppressive effect. The results of cytokine microarray showed that IL-17D concentration was at the highest level among inhibitory cytokines in the supernatant of γδ1 T cells from breast cancer tissues. The inhibitory effect of γδ1 T cells on the proliferation of CD4+ T cells could be reduced by anti-IL-17D monoclonal antibody. TLR8 ligand ssRNA40 inhibited the secretion of IL-17D, and then partially reversed the proliferating inhibition on the naive CD4+ T cells and immunosenescent induction by γδ1 T cells. Conclusion The γδ1 T cells derived from breast cancer tissues exert immunosuppressive effect by producing IL-17D to induce the immunosenescence of the naive CD4+ T cells. TLR8 ligand ssRNA40 can partially reduce the level of IL-17D secreted by γδ1 T cells, which can partially reverse the senescence and immunosuppression effect of γδ1 T cells on naive CD4+ T cells.

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Through gene mutation analysis of patients with non-syndromic hearing loss (NSHL) correct genetic counseling for patients with NSHL and their family members were provided. A total of 116 patients suffering from NSHL were selected, and Sanger sequencing was applied to analyze 31 mutation sites in four deafness genes [gap junction ß-2 (GJB2), solute carrier family 26, member 4 (SLC26A4), GJB3 and mitochondria 12S ribosomal ribonucleic acid (12SrRNA)]. Based on detection results, for the families with reproductive needs, amniotic fluid was extracted from pregnant women during proper gestational weeks to identify fetal genotypes and predict hearing state. Among 116 patients with NSHL, 51 patients carrying definite pathogenic mutation were found, including 35 patients with GJB2 mutations, 14 patients with SLC26A4 gene mutations and 2 patients with mitochondrial deoxyribonucleic acid 12SrRNA (mtDNA 12SrRNA) mutations. No GJB3 gene mutation site was detected. In addition, prenatal diagnosis to 17 pregnant women who had given birth to babies with deafness was performed, and results suggested that genotypes of 6 fetuses were consistent with those of probands, genotypes of 8 fetuses were consistent with those of their parents, and no mutation was found in the other 3 fetuses. Gene mutation analysis of patients with NSHL can identify the etiology and provide appropriate genetic counseling and birth guiding for patients with NSHL and their family members. In addition, prenatal diagnosis to the families who plan to give birth again can avoid the natality of fetuses with hearing loss.

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PURPOSE: Common single-nucleotide polymorphisms (SNPs) in microRNAs (miRs) have been shown to be associated with susceptibility to several types of human cancer. However, the association of miR-933 rs79402775 with gastric cancer (GC) has not been explored. METHODS: The association between rs79402775 in miR- 933 and the risk of GC was explored in Chinese population based on MassARRAY technology. A total 374 GC patients and 999 cancer-free controls were enrolled in this study. RESULTS: Compared with the wild-type GG, GA genotype was associated with a significantly decreased risk of GC (OR=0.542, 95% CI=0.299-0.983, p=0.044) in female subjects. Moreover, the variant was also associated with the expression of alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA). CONCLUSIONS: miR-933(rs79402775) may contribute to decreased susceptibility to GC and this SNP could be developed as a biomarker for GC prognosis.

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Although emerging evidence has indicated that single nucleotide polymorphisms (SNPs) in microRNAs (miRNAs) are associated with susceptibility to gastric cancer, a limited number of studies have revealed the underlying molecular mechanisms. In the present study, the results suggested that miR-1269a rs73239138 has a role in decreasing the risk of gastric cancer. The level of miR-1269a variant expression was significantly downregulated compared with the wild-type miR-1269a in the gastric cells (Fig. 1). Furthermore, overexpression of miR-1269a inhibited apoptosis of gastric cancer cells. Expression of the miR-1269a variant inhibited the function of miR-1269a by increasing the apoptotic rate and the expression of Bik, Bim and Bak was upregulated consistently. In addition, zinc-finger protein 70 (ZNF70) was identified to be a target gene of miR-1269a, which was downregulated by miR-1269a and upregulated by miR-1269a variant. ZNF70 was indicated to exert a role as a tumor suppressor in gastric cancer. To the best our knowledge, the present study for the first time highlights a critical role of miR-1269a variant rs73239138 in decreasing the susceptibility to gastric cancer by downregulating its expression and targeting ZNF70, which promotes apoptosis of gastric cancer cells. This SNP is indicated to serve as a potential biomarker and therapeutic target for gastric cancer.

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Colorectal cancer (CRC) is one of the most frequently occurring malignancies worldwide. The outcomes of patients with similar clinical symptoms or at similar pathological stages remain unpredictable. This inherent clinical diversity is most likely due to the genetic heterogeneity. The present study aimed to create a predicting tool to evaluate patient survival based on genetic profile. Firstly, three Gene Expression Omnibus (GEO) datasets (GSE9348, GSE44076 and GSE44861) were utilized to identify and validate differentially expressed genes (DEGs) in CRC. The GSE14333 dataset containing survival information was then introduced in order to screen and verify prognosis-associated genes. Of the 66 DEGs, the present study screened out 46 biomarkers closely associated to patient overall survival. By Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, it was demonstrated that these genes participated in multiple biological processes which were highly associated with cancer proliferation, drug-resistance and metastasis, thus further affecting patient survival. The five most important genes, MET proto-oncogene, receptor tyrosine kinase, carboxypeptidase M, serine hydroxymethyltransferase 2, guanylate cyclase activator 2B and sodium voltage-gated channel a subunit 9 were selected by a random survival forests algorithm, and were further made up to a linear risk score formula by multivariable cox regression. Finally, the present study tested and verified this risk score within three independent GEO datasets (GSE14333, GSE17536 and GSE29621), and observed that patients with a high risk score had a lower overall survival (P<0.05). Furthermore, this risk score was the most significant compared with other predicting factors including age and American Joint Committee on Cancer stage, in the model, and was able to predict patient survival independently and directly. The findings suggest that this survival associated DEGs-based risk score is a powerful and accurate prognostic tool and is promisingly implemented in a clinical setting.

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Single-nucleotide polymorphisms in microRNAs are related to the occurrence, development and prognosis of cancer. The aim of the present study is to investigate the possible influence of the miR-499(rs3746444) polymorphism on the risk of gastric cancer in Chinese population. A total of 363 GC patients and 969 cancer-free controls were enrolled in this study. The genotypes were obtained using MassARRAY method. The results showed that, compared with T allele, C allele was associated with a significantly increased risk of GC (OR=1.491, 95% CI=1.155-1.923, P=0.002). Moreover, a significantly increased risk of GC in subjects with the TC genotype was observed (adjusted OR of 1.559, 95% CI=1.148-2.117, P=0.004), compared with the wide type TT. We also found that basically dominant model (TT vs. TC+CC) was suitable for the association between rs6513497 and the risk of GC (OR=1.568, 95% CI=1.173-2.097, P=0.002). However, the same association was also shown in males and females. Meanwhile, rs3746444 was associated with the tumor size of GC patients. The present study indicated that miR-499 rs3746444 might contribute to GC risk and this SNP could be developed as a biomarker for GC prediction.

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SUMOylation is a post-translational modification exerted various effects on the target proteins. SUMOylation is a highly dynamic and reversible process, which has been shown to play an important role in tumorigenesis. However, the roles of sentrin/SUMO-specific proteases (SENPs), which mediate the reverse process of SUMOylation, in tumorigenesis remains largely unexplored. Here, we uncover a critical role of SENP6 in promoting gastric cancer cells growth via regulating the deSUMOylation of a transcription factor forkhead box protein M1 (FoxM1). We demonstrated that the mRNA and protein levels were elevated in gastric cancer tissues. Overexpression of SENP6 promoted, while RNA interference depletion of endogenous SENP6 inhibited gastric cancer cells growth and the ability of colony formation. By using biochemical assays, we identified FoxM1 as a novel substrate of SENP6 in gastric cancer cells. Thus, our data suggest that SENP6, which is highly expressed in gastric cancer cells, regulates the transcriptional activity and stability of FoxM1 through deSUMOylation.

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OBJECTIVE: To explore the clinical values of detecting drug related molecules excision repair cross complementing 1 (ERCC1) and top-isomerase I (TOPO I) in individualized therapies of metastatic colorectal cancer. METHODS: From June 2009 to December 2011, 90 patients at Huadong Hospital with metastatic colorectal cancer were randomly separated into 2 groups after operation. Each group had 45 patients without difference in gender, age or TNM stage. The expressions of ERCC1 and TOPO Iin cancer tissues were detected by immunohistochemical staining. The testing group received individualized chemotherapies following the expression results while the control group had random chemotherapies. The survival difference between two groups was analyzed by log-rank test and Kaplan-Meier analysis. And curative effect was analyzed by χ(2) or Fisher's analysis. RESULTS: The expressions of ERCC1 and TOPO I had no statistical significance between two groups (both P > 0.05). In the testing group, the median survival time was 281 days and the beneficial ratio 51.1% (23/45) versus 246 days and 44.4% (20/45) respectively in the control group. The inter-group comparisons of survival (P = 0.235) and curative effect (χ(2) = 0.04, P > 0.05) showed no statistical significance. In the estimated drug tolerated group (ERCC1 high expression or TOPO I low expression), the median survival time was 196 days and the beneficial ratio 4/14 versus 304 days and 51.3% (39/76) in the estimated drug sensitive group. The inter-group comparisons of survival and curative effect (both P < 0.05) had statistical significance. The median survival time and beneficial ratio significantly increased in estimated drug sensitive group than those in estimated drug tolerated group. CONCLUSIONS: The expression of drug related molecule in colorectal cancer tissue is significantly associated with curative effect in patients. Patients with down-regulated ERCC1 on Oxaliplatin or up-regulated TOPO Ion Irinotecan have longer survival and better curative effect. And chemotherapies guided by drug related molecule detection may boost curative effects in metastatic colorectal cancer.

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OBJECTIVE: To explore the relationships between the expression of transgelin in dendritic cells (DCs) pulsed with hepatocellular carcinoma lysates and the functions of the DCs. METHODS: DCs derived from healthy human white blood cells were divided into 3 groups: one was pulsed with high metastatic potential hepatocellular carcinoma cell line (MHCC97H) lysates, one with lysates of a low metastatic potential cell line (MHCC97L), and one un-pulsed DCs served as the control. The morphology of the DCs was observed by confocal microscopy and scanning electron microscopy. The phenotypes of the DCs were detected by flowcytometric analysis. The mixed leucocyte reaction (MLR) test and IL-12 secretion of DCs in the supernatants of MLR were employed to determine the functions of the DCs; the expression of transgelin was detected by Western blot. RESULTS: There were no morphological changes in the different DCs, but the levels of HLA-DR, CD80, CD83, CD86, MLR and IL-12 and transgelin were significantly higher in the two pulsed groups than those in the control group (P less than 0.01). In MHCC97H pulsed DCs, their CD80, CD83, CD86, and the expression of transgelin were also higher than those in the control group (P less than 0.05). The expression of transgelin was significantly higher in the MHCC97H pulsed group than in the MHCC97L loaded group, but CD80, CD83, CD86 and the level of IL-12 were all lower in the MHCC97H loaded DC group in comparison with those in the MHCC97 pulsed group (P less than 0.05). CONCLUSION: The expression of transgelin in DCs pulsed with HCC lysates is related to the functions of the DCs.

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PURPOSE: Serum low molecular weight protein biomarkers might be important in relation to portal vein tumor thrombi (PVTT) in hepatocellular carcinoma (HCC). This study aimed to screen and to detect these biomarkers. METHODS: We selected sera of 3 groups from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT, respectively. By using two-dimensional gel electrophoresis (2-DE) in which the first dimension was 16% SDS-PAGE, serum protein images of 3 groups were analyzed by Image Master Software. The differential protein spots were further identified by MALDI-TOF MS/MS. RESULTS: Compared with 12.5% SDS-PAGE gel, there were more protein bands between 3 and 20 kDa in 16% SDS-PAGE gel and low molecular weight (MW) protein spots (< 20 kDa) were clearly shown. Fifteen differential protein spots representing five proteins were found in the three groups by inter-class comparison and were then identified. Compared with the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were down regulated in HCC groups while haptoglobin-2 was over expressed. All the five proteins were less in PVTT group than in non-PVTT group. CONCLUSION: The expression of low MW serum protein changes obviously in the beginning and progressive stage of HCC, and differentially expressed low MW proteins might be the potential biomarkers in early prognostication and surveillance of treatment for HCC and PVTT.

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OBJECTIVE: To screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients. METHODS: Serum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software. The differential protein spots were further identified by MALDI-TOF MS/MS. RESULTS: Comparing the results using 12.5% SDS-PAGE gel, there were more protein bands (between 3 x 10(3) and 20 x 10(3)) and low molecular weight (MW) protein spots (less than 20 x 10(3)) were clearly shown in the 16% SDS-PAGE gel. Fifteen differential protein spots representing 5 proteins were found in the 3 groups by inter-class comparison and they were then identified. Compared with those in the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were all down regulated in HCC groups and haptoglobin-2 was over expressed. All 5 proteins decreased more in the PVTT group than in the non-PVTT group. CONCLUSION: The expression of low MW serum protein obviously changes in the beginning and in the progressive stage of HCC, and differentially expressed low MW proteins might be potential biomarkers in an early prognostic prediction and surveillance in the treatment for HCC and PVTT.

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