[Novel frameshift mutations in SALL4 in two Chinese families with Okihiro syndrome].

In the present study, clinical manifestations of two Chinese Okihiro syndrome families were analyzed, and genetic detections were performed on the two probands by exome sequencing and verified by Sanger sequencing for family members to determine the biological pathogenesis. Prenatal diagnoses were provided for three high-risk fetuses. The affected members exhibited a wildly spectrum of phenotypes, including ultrasound abnormalities of skeletal system (radius deformity and abnormal posture), and cardiac system (persistent common arterial trunk and ventricular septal defect) in the prenatal period of family 1, the severe phenotypes (grossly shortened and deformed forearm, Duane's anomaly and hearing loss), and the mild ones (usually only thenar dysplasia, or short radius styloid process). Two SALL4 variants, c.844delC p.(Q282Kfs*8) and c.2210delG p.(G737Vfs*23), have been identified respectively in two probands, and c.2210delG of SALL4 gene was unreported previously. The two variants were verified in all affected individuals, not in normal family members. Genotyping results of three fetuses indicated that one fetus was normal, and the two fetuses with heterozygous variation were affected. The two variants of SALL4 gene, c.844delC p.(Q282Kfs*8) and c.2210delG p.(G737Vfs*23), were the molecular pathological cause of Okihiro syndrome in the present study and enriched the spectrum of SALL4 variants. Our study provides accurate prenatal genetic diagnosis for the two families to avoid the birth of affected children.

[Analysis of the factors influencing positive predictive value of noninvasive prenatal testing for chromosome aneuploidies].

Objective: To investigate the influence of Z-score and different risk factors on positive predictive value (PPV) of noninvasive prenatal testing (NIPT) for chromosome aneuploidies. Methods: A total of 81 838 NIPT samples from January 1, 2016 to May 31, 2021 in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. Invasive prenatal diagnosis was applied to verify the diagnosis of NIPT-positive results and the corresponding PPV was calculated. The PPV of the samples with different Z-score were compared. The women were divided into high-risk group and non-high-risk group: high-risk group (n=39 114) included those with ultrasound soft index abnormalities, advanced maternal age or high risk for maternal serum screening, while non-high-risk group (n=42 724) included those with intermediate risk for maternal serum screening or no indications. The differences of the PPV between these two groups were compared. Finally, the comprehensive influence of Z-score and different risk factors on PPV were analyzed. Results: A total of 471 high-risk cases were detected by NIPT results, including 362 cases of trisomy 21, 77 cases of trisomy 18 and 32 cases of trisomy 13. For trisomy 21, trisomy 18 and trisomy 13, there were 226 cases, 46 cases and 6 cases which were confirmed via invasive prenatal diagnosis respectively. The corresponding PPV were 79.3% (226/285), 82.1% (46/56) and 27.3% (6/22), respectively. PPV of trisomy 21 and trisomy 18 were positively correlated with the corresponding Z-score (r=0.92, 0.62, all P<0.05), while trisomy 13 could not be analyzed due to the small sample size. The PPV of high-risk group was 85.2% (207/243), which was higher than that of the non-high-risk group with PPV of 59.2%(71/120, χ2=30.30, P<0.01). When the Z-score was between 3-<4 and 4-<5, the PPV of the high-risk group were 46.2%(12/26)and 62.5%(15/24) respectively, which were higher than those of the non-high-risk group [16.0%(4/25) and 14.3%(3/21), χ2=4.10, 8.90, all P<0.05]. With the increase of Z-score, there was no significant difference in PPV between the two groups (all P>0.05). Conclusions: The PPV of trisomy 21 and trisomy 18 are positively correlated with Z-score. The PPV of high-risk group is higher than that of non-high-risk group. The combination of Z-score and other risk factors may provide more accurate genetic counseling for those with NIPT positive results.

[Variation analysis of genes associated with Usher syndrome type 1 in 136 Chinese deafness families].

Objective: To investigate the variation of genes associated with Usher syndrome type 1(USH1)in 136 Chinese deafness families from Henan province. Methods: The data of 136 deafness families tested by next-generation sequencing(NGS) which identified in the center of genetics and prenatal diagnosis of the First Affiliated Hospital of Zhengzhou University from November 2016 to December 2019 were analysized and the variation frequency of six genes related to Usher syndrome type 1(MYO7A, USH1C, CDH23, PCDH15, USH1G, CIB2) were summarized. Results: Five deafness families were detected nine pathogenic or likely pathogenic variations in two genes, accounting for 3.7% of all families. Among them, four families were caused by MYO7A variations and one family was caused by CDH23 variation. Meanwhile, seven variations of two genes were reported for the first time. They were c.313delG, c.5257dupA, c.5435A>T, c.5636G>C, c.5722T>G of MYO7A, and c.155_166del, c.4802delA of CDH23. The patients' vision of family 2 and family 3 had no obvious abnormality at present, but according to genetic diagnosis and walking dealy, they were considered to be USH1. Conclusions: MYO7A is the most common caustive gene associated with USH1 in Henan deafness patients, the application of next-generation sequencing technology can make USH1 patients diagnosed earlier before the visual symptoms appear.

[Analysis of MYO15A variation in children with DFNB3].

Objective: To analyze the genetic and clinical characteristics of MYO15A variants associated non-syndromic autosomal recessive deafness3 (DFNB3). Methods: The hearing test and high-throughput sequencing data of 108 families with non-syndromic hearing loss, who visited the Center of Genetics and Prenatal Diagnosis in the First Affiliated Hospital of Zhengzhou University from November 2016 to February 2019, were retrospectively analyzed to investigate the characteristics of MYO15A variation. Results: Compound heterozygous MYO15A variations were detected in nine patients from eight families, accounting for 7.4% of all 108 families. The variants were c.5910+1G>A/c.9417_9418insTA, c.4234T>G/c.8324G>T, c.3926A>T/c.5002delC, c.9690+1G>A/c.10257_10259delCTT, c.8324G>T/c.10419_10423delCAGCT, c.4519C>T/c.6454G>C, c.6177+1G>T/c.10257_10259delCTT and c.5692C>T/c.7396-1G>A. All patients had severe to profound hearing loss. Among the 14 variations, 12 variations were located in the main structural domains, including 5 in motor domain, 3 in FERM domain, 3 in MyTH4 domain and 1 in IQ motif. The c.3926A>T, c.4234T>G, c.4519C>T, c.5002delC, c.6454G>C, c.8324G>T, c.9417_9418insTA and c.10419_10423delCAGCT had not been reported in the Human Gene Mutation Database up to February 2020. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG), 6 reported variants and the first reported c.4519C>T, c.5002delC, c.9417_9418insTA and c.10419_10423delCAGCT were identified as pathogenic variants, while c.8324G>T was likely pathogenic variant, and c.3926A>T, c.4234T>G and c.6454G>C were variants of uncertain significance. Conclusions: The variations of MYO15A in patients with DFNB3 are mainly complex heterozygous. The clinical phenotypes are mostly severe to profound hearing loss, and the mutation loci are mainly in the motor, FERM and MyTH4 domains.

[Analysis of genetic variation in patients with Waardenburg syndrome type â ¡ by next generation sequencing].

Objective: To detect gene mutation sassociated with deafness in four Waardenburg syndrome (WS) type Ⅱ patients, and to explore the possible mechanism of molecular genetics. Methods: All patients with WS were identified at the genetic and prenatal diagnosis center of the First Affiliated Hospital of Zhengzhou University from August 2015 to December 2018.Clinical materials and peripheral blood were collected from patients and family members. The genes associated with deafness of the patients were tested by next generation sequencing(NGS). And suspected mutations were verified by Sanger sequencing. Results: All patients carried heterozygous mutations in SOX10, they were c.355_356insTCAGGCAGCGC, c.1106_1107insTGGGGCCCCCCACACTA, c.511T>C (p.Y171H), c.91_100del. According to the guidelines for genetic variation of the Amercian College of Medical Genetics and Genomics (ACMG), three frameshift mutations were pathogenic mutations, one missense mutation was likely pathogenic mutation. Conclusion: Application of next generation sequencing technologies make gene diagnosis of Waardenburg syndrome efficiently and accurately.

MicroRNA-16 suppressed the invasion and migration of osteosarcoma by directly inhibiting RAB23.

OBJECTIVE: Osteosarcoma is the most frequent primary bone malignancy that affects young adults and adolescents around the world. Increasing evidence suggests that dysfunctions of microRNAs (miRNAs) used to play an important role in human cancers. We aimed at evaluating the potential function of miR-16 and verify its influence on the function of RAB23 in osteosarcoma. PATIENTS AND METHODS: miR-16 expressions in osteosarcoma tissues and cell lines were examined using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Transwell chambers were conducted to detect the miR-16 effects on osteosarcoma cells migration and invasion. Meanwhile, Western blot and luciferase assays were performed to validate RAB23 as miR-16 targets. RESULTS: miR-16 was down-regulated in osteosarcoma cell lines (MG63, SAOS-2, U2OS, and SOSP-9607) and osteosarcoma specimens, while RAB23 expression was higher in tumor tissues. Ectopic over-expression of miR-216 in osteosarcoma cells could inhibit cells migration and invasion. RAB23 was confirmed as a direct target of miR-16 and the inverse relationship between them was also observed. Over-expression of RAB23 ablates the inhibitory effects of miR-16. CONCLUSIONS: miR-16 inhibited cancer migration and invasion, and promoted the RAB23 expression in osteosarcoma. This newly identified miR-16/RAB23 axis may provide new insight into the pathogenesis and represents a potential therapeutic target for osteosarcoma.

[Clinical study on post-operational anti-metastasis effect of xiaoliu pingyi mixture].

OBJECTIVE: To observe the effect of combined use of chemotherapy (CT) and Xiaoliu Pingyi Mixture (XLPYM) in anti-metastasis, anti-relapse and tumor inhibition in patients after surgical operation. METHODS: Nienty-six patients of common tumor after operation were divided into 2 groups, the 58 patients in the observed group treated with CT plus XLPYM and the 38 patients in the control group treated with CT alone. After 2 courses of treatment, the status of 1 year remote metastasis and in local relapse of tumor were observed. For patients of relapse or metastasis, another 2 courses of treatment was given, then the changes of tumor size, quality of life, body weight, peripheral blood cells, immunity and hemorrheology were estimated and compared between the two groups. RESULTS: The remote metastatic rate and local relapse rate in the observed group were lower than those in the control group significantly (8.62% vs 44.74% and 15.52% vs 60.53%, P < 0.01). For patients of metastasis or relapse, the effective rate in the observed group was 46.24% and that in the control group was 24.00%, showing significant difference between the two groups (P < 0.05). Clinical observation also displayed that XLPYM could elevate the quality of life, regulate the disordered T-lymphocyte subsets, improve the hemorrheologic character and reduce the toxic-adverse reaction of CT in patients. CONCLUSION: XLPYM in combining with CT has effects of tumor inhibition, anti-metastasis and anti-relapse in treating patients of common tumor in post-operational period.