[Whole exome sequencing analysis and prenatal diagnosis in children with neurodevelopmental disorders].

To explore the application value of whole exome sequencing (WES) in the diagnosis of prenatal and postnatal neurodevelopmental disorders (NDDs). A total of 70 patients diagnosed with NDDs who underwent WES at the Medical Genetics Center of the Maternal and Child Health Hospital of Hubei Province between June 2020 and July 2021 were retrospectively analyzed. Genomic DNA was extracted from peripheral blood samples and amniotic fluid. WES-based copy number variant (CNV) analysis was integrated into the routine WES data analysis pipeline. The results showed that a molecular diagnosis rate could be made in 21/70 (30%) cases. Of 21 positive cases, 14 (23%) cases were detected by single-nucleotide variant/small insertion/deletion (SNV/Indel) analysis, of which 12 variants were novel, 6 (9.8%) cases were detected by WES-based CNV analysis, and 1 (1.6%) case was detected by a combination of both. The diagnostic yield of WES combined with CNV analysis was higher than that of SNV/Indel analysis alone (30%, 21/70 vs. 20%, 14/70). Of the 28 prenatally diagnosed cases, 6 cases were found to have inherited parental variation for NDDs, 10 cases were found not to have the same pathogenic variation as the proband, and the remaining 12 cases were found to have no pathogenic or likely pathogenic variation that could explain the NDDs phenotype. Clinical follow-up showed that 5 families opted for abortion and the remaining had no current abnormalities. In conclusion, WES may be an effective method to clarify the genetic etiology and prenatal diagnosis of NDDs, which is helpful in assessing the prognosis to aid clinical management and reproductive guidance.

[Correlation analysis of smell and taste loss with COVID-19 outbreak trend based on big data of internet].

Objective: To analyze the correlation between loss of smell/taste and the number of real confirmed cases of coronavirus disease 2019 (COVID-19) worldwide based on Google Trends data, and to explore the guiding role of smell/taste loss for the COVID-19 prevention and control. Methods: "Loss of smell" and "loss of taste" related keywords were searched in the Google Trends platform, the data were obtained from Jan. 1 2019 to Jul. 11 2021. The daily and newly confirmed COVID-19 case number were collected from World Health Organization (WHO) since Dec. 30 2019. All data were statistically analyzed by SPSS 23.0 software. The correlation was finally tested by Spearman correlation analysis. Results: A total of data from 80 weeks were collected. The retrospective analysis was performed on the new trend of COVID-19 confirmed cases in a total of 186 292 441 cases worldwide. Since the epidemic of COVID-19 was recorded on the WHO website, the relative searches related to loss of smell/taste in the Google Trends platform had been increasing globally. The global relative search volumes of "loss of smell" and "loss of taste" on Google Trends was 10.23±2.58 and 16.33±2.47 before the record of epidemic while 80.25±39.81 and 80.45±40.04 after (t value was 8.67, 14.43, respectively, both P<0.001). In the United States and India, the relative searches for "loss of smell" and "loss of taste" after the record of epidemic were also much higher than before (all P<0.001). The correlation coefficients between the trend of weekly new COVID-19 cases and the Google Trends of "loss of smell" in the global, United States, and India was 0.53, 0.76, and 0.82 respectively (all P<0.001), the correlation coefficients with Google Trends of "loss of taste" was 0.54, 0.78, and 0.82 respectively (all P<0.001). The lowest and highest point of loss of smell/taste search curves of Google Trends in different periods appeared 7 to 14 days earlier than that of the weekly newly COVID-19 confirmed cases curves, respectively. Conclusions: There is a significant positive correlation between the number of newly confirmed cases of COVID-19 worldwide and the amount of keywords, such as "loss of smell" and "loss of taste", retrieved in Google Trends. The trend of big data based on Google Trends might predict the outbreak trend of COVID-19 in advance.

Development and characterization of microsatellite markers of the eastern keelback mullet (Liza affinis).

Twenty-four polymorphic microsatellite loci were isolated and characterized for Liza affinis using a (GT)13-enriched genomic library. The number of alleles per locus ranged from 3 to 9, with a mean number of 6.250. The observed and expected heterozygosities ranged from 0.417 to 1.000 and from 0.550 to 0.861, with an average of 0.859 and 0.779, respectively. Deviation from Hardy-Weinberg proportions was detected at three loci. Evidence of null alleles was found at two loci. These markers will be useful in further studies investigating the genetic variation and population structure of this species, and may provide insights into the maintenance and efficient management of eastern keelback mullet resources.

Development of polymorphic microsatellite loci for a new fish species, Chinese sillago (Sillago sinica).

Sillago sinica is a newly identified species belonging to Sillaginidae, Perciforms, and was found along the coast of China in 2011. In the present study, 81 microsatellite loci were isolated from an enriched genomic library, and 24 positive clones containing microsatellite repeats had adequate flanking sequences for the development of PCR primers. Sixteen of these primers were monomorphic or would not amplify. Eight were polymorphic in an examined population with the number of alleles per locus ranging from 2 to 14. The number of observed and expected heterozygosities per locus varied from 0.125 to 0.958 and from 0.120 to 0.904, respectively. The polymorphism information content ranged from 0.110 to 0.721. All loci conformed to Hardy-Weinberg equilibrium (P > 0.05) after Bonferroni correction. There was no significant linkage disequilibrium between the eight polymorphic loci. These results suggest that these markers may be very useful for the characterization of natural populations of this species.

[Study on IGF-1 and IGFBP-3 gene expression in liver of young rats with intermittent hypoxia].

Objective:Using the model of young rats with intermittent hypoxia, to study the expression of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) in liver, and to provide objective evidence for the possible existence of growth retardation in children with OSAHS. Method:Divided 24 rats randomly into three groups, as control group and the mild and severe hypoxia group. Normally fed control group, without treatment, the other two groups were kept in the cabin has been simulated mild and severe intermittent hypoxia conditions of OSAHS in children. 8-hour day cycle of intermittent hypoxia, a total of 35 days. To the last day, animals were sacrificed and fresh liver tissues were fixed in liquid nitrogen, stored in the -80°C refrigerator, then took the molecular biology experiment of IGF-1 and IGFBP-3 in the liver. Result:The expression of IGF-1 mRNA in liver, semi-quantitative analysis showed that the difference of mRNA expression of IGF-1 among the three groups was statistically significant (P<0.05), pairwise comparison, mild hypoxia group was higher than control and severe group, there were significant differences (P<0.05), the control group and severe hypoxia had no significant difference (P>0.05). The expression of IGFBP-3 mRNA in liver showed that, differences between the three groups was not significant (P>0.05). IGF-1 expression in liver tissue analysis showed that, differences between the three groups was statistically significant (P<0.05), pairwise comparisons, its expression in the control group was lower than mild hypoxia group and severe group, there were significant differences (P<0.05), mild hypoxia and severe hypoxia group had no significant difference (P>0.05). IGFBP-3 expression in liver tissue analysis showed that, differences between the three groups was statistically significant (P<0.05), pairwise comparisons, the control group was higher than mild and severe hypoxic group, there were significant differences (P<0.05), mild hypoxia and severe hypoxia group had no significant difference (P>0.05). Conclusion:IGF-1 and IGFBP-3 in the liver of mild hypoxia group, severe hypoxic group and control group showed that their transcription levels and the severity of hypoxia had no significant correlation.

In vitro proliferation and differentiation of hepatic oval cells and their potential capacity for intrahepatic transplantation.

Hepatic oval cells (HOCs) are recognized as facultative liver progenitor cells that play a role in liver regeneration after acute liver injury. Here, we investigated the in vitro proliferation and differentiation characteristics of HOCs in order to explore their potential capacity for intrahepatic transplantation. Clusters or scattered HOCs were detected in the portal area and interlobular bile duct in the liver of rats subjected to the modified 2-acetylaminofluorene and partial hepatectomy method. Isolated HOCs were positive for c-kit and CD90 staining (99.8% and 88.8%, respectively), and negative for CD34 staining (3.6%) as shown by immunostaining and flow cytometric analysis. In addition, HOCs could be differentiated into hepatocytes and bile duct epithelial cells after leukemia inhibitory factor deprivation. A two-cuff technique was used for orthotopic liver transplantation, and HOCs were subsequently transplanted into recipients. Biochemical indicators of liver function were assessed 4 weeks after transplantation. HOC transplantation significantly prolonged the median survival time and improved the liver function of rats receiving HOCs compared to controls (P = 0.003, Student t-test). Administration of HOCs to rats also receiving liver transplantation significantly reduced acute allograft rejection compared to control liver transplant rats 3 weeks following transplantation (rejection activity index score: control = 6.3 ± 0.9; HOC = 3.5 ± 1.5; P = 0.005). These results indicate that HOCs may be useful in therapeutic liver regeneration after orthotopic liver transplantation.

In vitro proliferation and differentiation of hepatic oval cells and their potential capacity for intrahepatic transplantation

Hepatic oval cells (HOCs) are recognized as facultative liver progenitor cells that play a role in liver regeneration after acute liver injury. Here, we investigated the in vitro proliferation and differentiation characteristics of HOCs in order to explore their potential capacity for intrahepatic transplantation. Clusters or scattered HOCs were detected in the portal area and interlobular bile duct in the liver of rats subjected to the modified 2-acetylaminofluorene and partial hepatectomy method. Isolated HOCs were positive for c-kit and CD90 staining (99.8% and 88.8%, respectively), and negative for CD34 staining (3.6%) as shown by immunostaining and flow cytometric analysis. In addition, HOCs could be differentiated into hepatocytes and bile duct epithelial cells after leukemia inhibitory factor deprivation. A two-cuff technique was used for orthotopic liver transplantation, and HOCs were subsequently transplanted into recipients. Biochemical indicators of liver function were assessed 4 weeks after transplantation. HOC transplantation significantly prolonged the median survival time and improved the liver function of rats receiving HOCs compared to controls (P=0.003, Student t-test). Administration of HOCs to rats also receiving liver transplantation significantly reduced acute allograft rejection compared to control liver transplant rats 3 weeks following transplantation (rejection activity index score: control=6.3±0.9; HOC=3.5±1.5; P=0.005). These results indicate that HOCs may be useful in therapeutic liver regeneration after orthotopic liver transplantation.

Monolayer polystyrene micro-spheres array master derived by spin-coating method for UV nanoimprint.

Polystyrene micro-spheres aqueous suspensions were obtained by an emulsifier-free polymerization method. Fourier transform infrared spectroscopy and laser particle size distribution were used to characterize the colloidal solutions. Results indicate that the polystyrene micro-spheres colloidal solution has a good dispersion and the micro-spheres have a uniform size and smooth surface. The as-prepared polystyrene micro-spheres colloidal solution was spun onto glass substrates and then the coated sample was heated at 70 degrees C for 1 h to form close-packed monolayer polystyrene microspheres arrays, which are used as a master. Scanning electron microscopy was used to observe the morphological properties of the polystyrene micro-spheres array. A relative good result can be easily obtained by a spinning speed of 2000 rpm for 30 second, when the solution concentration is around 10 wt.%. Furthermore, the polydimethylsiloxane soft mold was replicated from the-fabricated polystyrene micro-spheres array master and was then imprinted onto the photosensitive hybrid film under UV-irradiation and thus the micro-lens arrays built in the photosensitive hybrid sol-gel film were obtained.

Remarkably low mtDNA control-region diversity and shallow population structure in Pacific cod Gadus macrocephalus.

To investigate the genetic diversity and describe the population structure in Gadus macrocephalus, a 452 base pair (bp) fragment of the mitochondrial DNA control region was analysed in 259 individuals. The results showed remarkably low nucleotide diversity and a lack of genealogical structure. Small but significant genetic differentiations, however, were detected among north-western Pacific populations, but no large-scale regional differences were detected. These results indicate that populations of G. macrocephalus in the north-western Pacific are genetically subdivided and represent evolutionary lineages that should be managed individually.

Population genetic structure and larval dispersal potential of spottedtail goby Synechogobius ommaturus in the north-west Pacific.

Larval dispersal may have an important effect on genetic structure of benthic fishes. To examine the population genetic structure of spottedtail goby Synechogobius ommaturus, a 478 base pair (bp) fragment of the hypervariable portion of the mtDNA control region was sequenced and used to interpret life-history characteristics and larval dispersal strategy. Individuals (n = 186) from 10 locations on the coasts of China and Korea were analysed and 44 haplotypes were obtained. The levels of haplotype and nucleotide diversity were higher in East China Sea populations than in other populations. Both the phylogenetic tree and the minimum spanning tree showed that no significant genealogical structures corresponding to sampling locations existed. AMOVA and pair-wise F(ST) revealed significant genetic differentiation between populations from Korea and China. A significant isolation by distance pattern was observed in this species (r = 0.53, P < 0.001). Both mismatch distribution analysis and neutrality tests showed S. ommaturus to have experienced a recent population expansion. These results suggest that the Pleistocene ice ages had a major effect on the phylogeographic pattern of S. ommaturus, that larvae might avoid offshore dispersal and that dispersal of larvae may maintain a migration-drift equilibrium.

In vivo induction of malignant transformation of nude mouse B lymphocytes by a human multiple myeloma.

The bone marrow puncture fluid of human multiple myeloma was inoculated subcutaneously in BALB/C/nu nude mice. After passage of the xenograft from nude mice in vitro, a murine malignant lymphosarcoma cell line was unexpectedly established, which was named NLB-SK. The murine NLB-SK cell line has been cultivated in vitro over 72 passages for 10 months. Repeated cryopreservation showed that the murine NLB-SK cell line revived satisfactorily. Based on cell biological characteristics the malignant transformation might be attributed to horizontal oncogenesis between human malignant tumor cells and nude mouse normal somatic cells.