OBJECTIVE: To explore the association between de novo mutations (DNM) and non-syndromic cleft lip with or without palate (NSCL/P) using case-parent trio design. METHODS: Whole-exome sequencing was conducted for twenty-two NSCL/P trios and Genome Analysis ToolKit (GATK) was used to identify DNM by comparing the alleles of the cases and their parents. Information of predictable functions was annotated to the locus with SnpEff. Enrichment analysis for DNM was conducted to test the difference between the actual number and the expected number of DNM, and to explore whether there were genes with more DNM than expected. NSCL/P-related genes indicated by previous studies with solid evidence were selected by literature reviewing. Protein-protein interactions analysis was conducted among the genes with protein-altering DNM and NSCL/P-related genes. R package "denovolyzeR" was used for the enrichment analysis (Bonferroni correction: P=0.05/n, n is the number of genes in the whole genome range). Protein-protein interactions among genes with DNM and genes with solid evidence on the risk factors of NSCL/P were predicted depending on the information provided by STRING database. RESULTS: A total of 339 908 SNPs were qualified for the subsequent analysis after quality control. The number of high confident DNM identified by GATK was 345. Among those DNM, forty-four DNM were missense mutations, one DNM was nonsense mutation, two DNM were splicing site mutations, twenty DNM were synonymous mutations and others were located in intron or intergenic regions. The results of enrichment analysis showed that the number of protein-altering DNM on the exome regions was larger than expected (P < 0.05), and five genes (KRTCAP2, HMCN2, ANKRD36C, ADGRL2 and DIPK2A) had more DNM than expected (P < 0.05/(2×19 618)). Protein-protein interaction analysis was conducted among forty-six genes with protein-altering DNM and thirteen genes associated with NSCL/P selected by literature reviewing. Six pairs of interactions occurred between the genes with DNM and known NSCL/P-related genes. The score measuring the confidence level of the predicted interaction between RGPD4 and SUMO1 was 0.868, which was higher than the scores for other pairs of genes. CONCLUSION Our study provided novel insights into the development of NSCL/P and demonstrated that functional analyses of genes carrying DNM were warranted to understand the genetic architecture of complex diseases.
Asians ; Case-Control Studies ; Cleft Lip/genetics* ; Cleft Palate/genetics* ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Genotype ; Humans ; Mutation ; Parents ; Polymorphism, Single Nucleotide ; Whole Exome Sequencing

Next-generation sequencing has revolutionized family-based association tests for rare variants. As the lower power of genome wide association study for detecting casual rare variants, methods aggregating effects of multiple variants have been proposed, such as burden tests and variance component tests. This paper summarizes the methods of rare variants association test that can be applied for family data, introduces their principles, characteristics and applicable conditions and discusses the shortcomings and the improvement of the present methods.
Computer Simulation ; Family Relations ; Genetic Association Studies ; Genetic Variation ; Genome-Wide Association Study/methods* ; Humans

Objective To analyze the sequences of mitochondrial cytochrome C oxidase subunit I gene (COI) and 18S ribosomal RNA gene (18S rRNA), so as to identify the feasible DNA barcodes for 4 species of cheyletid mites and improve the DNA barcoding database for cheyletid mites. Methods Cheyletid mite samples were collected from small-scale flour mills in Fuyang, Wuhu and Tongling cities of Anhui Province from May 2018 to July 2019, extracted and morphologically identified. Then, genomic DNA was extracted from a single cheyletid mite, and the COI and 18S rRNA gene sequences were obtained by PCR amplification, cloning and sequencing. The obtained sequences were aligned using the BLAST software. Multiple sequence alignment was done using the software ClustalX version 1.83 using the known gene sequences from cheyletid mites. The genetic distance was calculated using the software MEGA X, and the phylogenetic tree was created using the maximum likelihood method. Results The DNA barcoding results of Cheyletus malaccensis, C. carnifex and Cheletomorpha lepidopterorum were consistent with the morphological identification, while no sequences pertaining to Eucheyletia reticulate were retrieved in the GenBank database. The proportions of A + T were 69.6% and 55.1% in the COI and 18S rRNA sequences of 4 cheyletid mites species, respectively, and the numbers of base substitutions were 137 and 46, respectively. There were 154 to 321 and 58 to 99 inter-species variation loci in the COI and 18S rRNA gene sequences of 4 cheyletid mites species, respectively, and the intra-species genetic distance was all 0.020 or less in the COI and 18S rRNA gene sequences of 4 cheyletid mites species, with inter-species genetic distance of 0.235 to 0.583 and 0.078 to 0.114, respectively. Phylogenetic analysis based on COI and 18S rRNA genes showed that all four species of cheyletid mites were clustered into a branch with a 100% supportive rate, which was consistent with the morphological identification. Conclusion Mitochondrial COI gene is superior to 18S rRNA gene as DNA barcodes for 4 species of cheyletid mites, which is more suitable to be used to investigate the phylogenetic relationship of at genus and species levels.

Our study reports a case of acarodermatitis caused by Haemolaelaps casalis. By morphological observations, the mites seized were identified as Haemolaelaps casalis (deutonymph) which could attack humans resulting in acarodermatitis characterized with the symptoms of papules and blisters in different degrees. The patient was treated with 15% calamine lotion and anti-inflammatory and antipruritic drugs. Meanwhile, the mites were eliminated in the bedroom. After the treatment for one week, the patient was cured. Haemolaelaps casalis, which had been found in the indoor mattress, could attack humans and cause acarodermatitis. We should strengthen the work of anti-mite in domestic environment.

Objective To explore the fire prevention management to ensure the operating of the data room in the hospital. Methods The hidden risks in the fire prevention of the data room were pointed out in auto fire fighting system,precision air-conditioning equipment, UPS and battery pack, compatibilization and expansion, high-voltage power supply circuit, lightningproof grounding and etc.It's indicated that the maintenance staff had deficiency in mastering related knowledge on gas fire-extinguishing system.Results The maintenance had to be strengthened for the auto fire fighting system,precision air-conditioning equipment,UPS and battery pack,and the detection should be reinforced for the high-voltage power supply circuit and lightningproof grounding system. It's suggested that standardized construction be executed during data room compatibilization and expansion,corresponding management system be established,and the knowledge be mastered on the composition,operation and announcements.Conclusion The fire prevention management of the data room is enhanced in the hospital,and the hidden risks are eliminated for fire fighting.

OBJECTIVETo research the correlation between surgical treatment for atlantoaxial dislocation and dredging Governor vessel.

METHODSFrom June 2004 to June 2010,113 patients were reviewed, including 65 males and 48 females, with the mean age of 42.6 years (ranged, 8 to 64 years). All the patients were classified and treated by TOI clinical classification which included traction and decompression and reduction, and inter fixation fusion by surgery. The SAC (space available for the cord), Governor vessel Yuzu score, JOA score, NDI score (cervical spine dysfunction index) before treatment were compared with those of after treatment.

RESULTSThe SAC in each group before treatment were (13.34 +/- 3.21), (10.43 +/- 2.42), (7.89 +/- 3.14), (10.50 +/- 0.71) mm respectively, the last follow-up of (16.02 +/- 1.42), (15.34 +/- 1.87), (14.49 +/- 1.58), (12.50 +/- 0.71) mm; Governor vessel Yuzu score before treatment were (8.37 +/- 1.87), (6.87 +/- 1.35), (5.17 +/- 1.13), (7.50 +/- 0.71) respectively, the last follow-up of (10.59 +/- 0.94), (10.25 +/- 1.01), (8.41 +/- 1.31), (9.0 +/- 0); JOA, NDI score significantly improved compared with that of before treatment.

CONCLUSIONAtlantoaxial dislocation confirmed and treated by TOI clinical classification can effectively relieve the nerve function and Governor vessel Yuzu syndrome, and show that surgical treatment is closely related with dredging the Governor vessel.

Adolescent ; Adult ; Atlanto-Axial Joint ; surgery ; Child ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Retrospective Studies

ObjectiveTo investigate the current status of Kaschin-Beck disease in Shandong province, and to provide a scientific basis for decision-making in controlling the disease. Methods According to the National Monitoring Program of Kaschin-Beck disease requirements, historical serious villages of Kaschin-Beck disease in Qingzhou of Shandong province were selected annually; children aged 7 to 16 were chosen to receive clinical examination and children aged 7 to 12 were taken X-ray examination. Clinical and X-ray diagnosis was carried out according to the Diagnostic Criteria of Kashin Beck Disease(GB 16003-1995). Results From 1996 to 2010, in 53 diseased villages, three thousand three hundred and eighteen school children aged 7 to 16 were clinically diagnosed, and child Kaschin-Beck disease of degree Ⅰ and above were not detected; three thousand and ninety-one school children aged 7 to 12 were examined by X-ray, forty cases were found positive, and the total positive rate was 1.29％(40/3091 ). The year with the highest positive rate was 2002, and the rate was 3.49％(13/372) ; the positive rate was 0 in 1996 and 2008. The difference of the X-ray positive rate between each year was statistically significant(x2 =31.54, P ＜ 0.01 ). ConclusionsChild Kashin-Beck disease in Qingzhou is basically under control.Since etiology of Kashin-Beck disease is still unclear, surveillance of the disease still needs to be strengthened.

Objective To explore the effect of polyethylene glycol-polyethyleneimine (PEG-PEI) serving as a non-viral vector in delivering small interfering RNA (siRNA) into C17.2 neural stem cells (NSCs) in vitro. Methods Complexes of PEG-PEI and siRNA targeting Nogo receptor were prepared, and their characterizations were estimated by measurements of particle size and zeta potential,and the complex abilities of PEG-PEI/siRNA complexes were observed by gel retardation assay. In addition, with liposome complex system (Lipofectamine 2000/siRNA) as positive control, the transfection efficiency of PEI-PEG/siRNA complexes at different N/P ratios (cationic nitrogen/siRNA phosphate molar ratio) was detected by flow cytometry. Results The siRNA molecules were condensed by PEG-PEI to form nanoseale complexes. As the proportion of N/P ratio enhancing, the surface potential of nanoparticles gradually increased and the particle sizes of PEI-PEG/siRNA complexes showed a decreasing trend. Gel retardation electrophoresis suggested that siRNA could be fully composited with PEG-PEI as a result of the coulombic foree between them. Meanwhile, flow cytometry experiments revealed that the transfection efficiency of PEG-PEI mainly depended on N/P ratios of the nanoparticles,and the highest one was obtained at N/P=15 ([78.72±8.18)]％). Conclusion PEG-PEI might be a prospective candidate for siRNA delivery system, which enjoys its value in NSC gene therapy.

OBJECTIVETo study the genetic aberrations of ocular extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type occurring in patients from southern China.

METHODSFifty seven paraffin-embedded ocular MALT lymphoma specimens from patients in southern China were studied by interphase fluorescence-in-situ hybridization (FISH) for genetic aberrations including t(11;18)(q21;q21)/API2-MALT1, t(1;14)(p22;q32)/IgH-bcl-10, t(14;18) (q32;q21)/IgH-MALT1 and bcl-6/FOXP1 gene translocations.

RESULTSAmongst the 57 cases studied, 9 cases (15.8%) showed chromosome translocations, including 4 cases (7.0%) of t(11;18)(q21;q21)/API2-MALT1, 1 case (1.8%) of t(14;18) (q32;q21)/IgH-MALT1, 1 case (1.8%) of bcl-6 gene-related chromosome translocation and 3 cases (5.3%) of IgH-unknown translocation partner. FISH revealed 17 cases (29.8%) with 3 copies of bcl-6 gene, 21 cases (36.8%) with 3 copies of MALT1 gene and 12 cases (21.1%) with 3 copies of both genes.

CONCLUSIONSThe MALT lymphoma-associated chromosome translocations t(11;18)(q21;q21)/API2-MALT1 and t(14;18) (q32;q21)/IgH-MALT1 are demonstrated in ocular MALT lymphomas of southern Chinese patients. The prevalence is significantly different from that reported in northern Chinese and northern American patients, indicating a geographic heterogeneity in the MALT lymphoma-associated genetic aberrations. The presence of 3 copies of bcl-6 and MALT1 genes is the commonest genetic abnormalities observed in ocular MALT lymphomas, suggesting a possible role in MALT lymphomagenesis.

Caspases ; genetics ; metabolism ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 11 ; genetics ; Chromosomes, Human, Pair 14 ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; Chromosomes, Human, Pair 3 ; genetics ; DNA-Binding Proteins ; genetics ; metabolism ; Eye Neoplasms ; genetics ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, B-Cell, Marginal Zone ; genetics ; metabolism ; Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein ; Neoplasm Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; Translocation, Genetic ; Trisomy

Objective To investigate the present status of Kaschin-Beck disease(KBD) in Qingzhou at present, and provide a scientific basis for decision-making in controlling the disease. Methods According to The National Technical Scheme for Endemic Disease Control in 2007, children aged 7 to 12 were chosen to receive clinical and X-ray examination in 3 villages of 2 towns in Qingzhou. Local adults aged 16 years and older were clinically diagnosed in 10 villages of 4 towns and divided into different groups according to the clinical degrees and their ages. The Diagnostic Criteria of Kashin Beck Disease(GB 16003-1995) was carded out by clinical and X-ray diagnosis. Results Two hundred and sixty school children aged 7 to 12 were clinically diagnosed, and the patients of I degree upwards of KBD were not detected. One hundred and ninety-eight school children aged 7 to 12 were examined by X-ray, and the detectable rate was 0(0/198). In 7099 adults aged 16 years old upwards clinically tested, 502 patients of degree I and higher of KBD were detected and the detectable rate was 7.07%(502/7099). The patients distributed mainly in the population aged 36 years old and older, accounting for 99.00% (497/502). Conclusions Although KBD in children have been controlled in Qingzhou, it is still serious in adults. So the monitoring of KBD should be carried on and prevention and control for KBD in adults should be strengthened.