Objective To use the matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) technique for detecting the mutation gene of neonatal non-syndromic hereditary hearing impairment gene in Guangxi and to investigate its effectiveness and feasibility in clinical application.Methods A total of 7 100 newborns were performed the hearing preliminary screening and secondary screening by adopting AABR.The genomic DNA was extracted by the heel blood spot.Twenty mutation characteristics of 4 deaf predisposing genes were detected by MALDI-TOF-MS.Results The pass rate of hearing screening in 7 100 newborns was 97.11％ (6 895/7 100),the positive rate of neonatal gene mutation was 3.54％ (251/7 100),in which the GJB2 gene mutation was in 131 cases,the carrying rate was 1.84％,235delC heterozygous mutation was in 108 cases.SLC26A4 gene mutation was in 93 cases,which dominated by 1229C＞T heterozygous mutation and IVS7-2A＞G heterozygous mutation,mtDNA12SRNA gene mutation was in 16 cases and GJB3 gene mutation was in 11 cases.Conclusion Adopting the MALDI-TOF-MS screening technique can increase the detection rate of hot point mutation in common deaf related genes and discover neonatal genetic NSHI from molecular level and provides the corresponding geneticconsulting guidance for early finding and predicting deaf occurrence,and formulating the interventional measures.

BACKGROUND:Previous studies discovered that pRST98, original y isolated from Salmonel a enterica serovar typhimurium (S.typhimurium) could promote bacterial biofilm formation. In addition, bacterial harboring pRST98 can promote the secretion and expression of interleukin-10 after infection in cells and animals.
OBJECTIVE:In vitro studies have discovered the effects of interleukin-10 at varying concentrations and conjugative plasmid pRST98 on the biofilm formation of S.typhimurium.
METHODS:S.typhimurium wild-type strainχ3306, virulence plasmid-deletion S.typhimurium strainχ3337 and pRST98-transconjugant S.typhimuriumχ3337/pRST98 were established in vitro and cultured for biofilm formation. 1, 10, 100 μg/L interleukin-10 were added during the biofilm formation. 0 μg/L interleukin-10 was set as a control. Crystal violet staining method, semi-quantitative method, confocal laser scanning microscopy and scanning electron microscopy were used to determine the effects of interleukin-10 on the biofilm formation and compare the effects of S.typhimurium with or without pRST98.
RESULTS AND CONCLUSION:Intra-group comparison showed that, compared with the control group, S.typhimurium gathered together and formed thicker biofilm in concentration of 1 and 10 μg/L of interleukin-10. The promotion effects of S.typhimurium on biofilm formation were greatly improved in 10 μg/L. Interleukin-10 in 100 μg/L inhibited S.typhimurium biofilm formation. Inter-group comparison showed that, A570 inχ3337/pRST98 was greatly higher than that inχ3306 andχ3337 under the same concentration of interleukin-10. The results indicate that both 1 and 10 μg/L of interleukin-10 promote biofilm formation, especial y bacteria harboring pRST98.

OBJECTIVE: To perform the validation and analysis of forensic parameters of Goldeneye DNA ID 26Y system. METHODS: Based on the validation rules of Scientific Working Group on DNA Analysis Methods (SWGDAM), the kit was assessed from several parts, as test of PCR system, reproducibility, accuracy, and sensitivity, etc. And Y-STR loci of 517 unrelated healthy individuals from Eastern China were genotypes by this kit. The distribution and frequency of haplotype were calculated and forensic parameters of the kit were assessed. RESULTS: The complete profiles can be obtained even when the PCR reaction volume with 6.25 microL. And correct profile was obtained with DNA down to 125 pg. No reproducible peaks were detected with the DNA of common animals and microorganism with the kit. For the male-male mixture testing, average 70% of the minor alleles were obtained when the ratios of 1:19 and 19:1. For the male-female mixture testing, results showed that the sensitivity of the kit was no compromised with the addition of female samples. CONCLUSION The validation studies demonstrated that Goldeneye DNA ID 26Y system has good sensitivity and specificity, and suitable for mixture testing. The polymorphism of 26 Y-STR loci included in this kit are good for forensic application.
Alleles ; Animals ; Asian People/genetics* ; China ; Chromosomes, Human, Y ; DNA ; DNA Fingerprinting/standards* ; Female ; Forensic Genetics/methods* ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVE: To investigate the genetic data of 21 autosomal STR included in Goldeneye™ DNA ID 22NC Kit in Chinese Han nationality and to evaluate the forensic application. METHODS: By detected 500 unrelated healthy individuals in Chinese Han nationality of East China with Goldeneye™ DNA ID 22NC Kit, allele frequencies, population genetics parameters and linkage disequilibrium information of the 21 autosomal STR were statistically analyzed. RESULTS: In the 21 autosomal STR, no deviations from Hardy-Weinberg equilibrium were detected and all loci were independent form each other. DP values of 21 autosomal STR were all above 0.85, and the combined discrimination power was 1-3.616 5 x 10(-26). Combined mean exclusion chance of this system in duo cases was 1-2.786 81 x10(-6), in trio cases was 1-8.545 82 x 10(-1). CONCLUSION Twenty-one autosomal STR included in Goldeneye™ DNA ID 22NC Kit are highly polymorphic in the Han nationality. Combined with Goldeneye™ DNA ID 20A Kit, the kit can satisfy the needs for full-sibling testing and facilitate the solution of this kind of case tools.
Alleles ; Asian People/genetics* ; China ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genetic Markers/genetics* ; Genetics, Population ; Genotype ; Humans ; Polymorphism, Genetic ; Reagent Kits, Diagnostic

Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

OBJECTIVE: To evaluate the power of Identifiler System for paternity testing. METHODS: A total of 3 277 paternity testing cases were studied using Identifiler System. The exclusion power and mutation rates of the Identifiler System were analysed in the paternity testing. RESULTS: The cumulated power of exclusion was 0.999 998 827, and the cumulated discriminating power was 0.999 999 999 999 999 98, respectively. Of the 3 277 cases, paternity was confirmed in 2 863, but excluded in 347. Among this paternity testing, mutations involving a single STR locus were observed in 65 cases, while mutations involving 2 STR loci were observed in 2 cases. CONCLUSION The Identifiler System is powerful and reliable for paternity testing.
Alleles ; China ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Genetic Testing/methods* ; Genetics, Population ; Humans ; Microsatellite Repeats ; Mutation ; Paternity ; Polymerase Chain Reaction/methods* ; Probability ; Tandem Repeat Sequences/genetics*

OBJECTIVE: Determination strategies for half sibling sharing a same mother were investigated through the detection of autosomal and X-chromosomal STR (X-STR) loci and polymorphisms on hypervariable (HV) region of mitochondrial DNA (mtDNA). METHODS: Genomic DNA were extracted from blood stain samples of the 3 full siblings and one dubious half sibling sharing the same mother with them. Fifteen autosomal STR loci were genotyped by Sinofiler kit, and 19 X-STR loci were genotyped by Mentype Argus X-8 kit and 16 plex in-house system. Polymorphisms of mtDNA HV-I and HV-II were also detected with sequencing technology. RESULTS: Full sibling relationship between the dubious half sibling and each of the 3 full siblings were excluded based on the results of autosomal STR genotyping and calculation of full sibling index (FSI) and half sibling index (HIS). Results of sequencing for mtDNA HV-I and HV-II showed that all of the 4 samples came from a same maternal line. X-STR genotyping results determined that the dubious half sibling shared a same mother with the 3 full siblings. CONCLUSION It is reliable to combine three different genotyping technologies including autosomal STR, X-STR and sequencing of mtDNA HV-I and HV-II for determination of half sibling sharing a same mother.
Chromosomes, Human, X/genetics* ; DNA, Mitochondrial/genetics* ; Female ; Forensic Genetics/methods* ; Genetic Markers ; Genotype ; Humans ; Male ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Siblings ; Tandem Repeat Sequences/genetics*

OBJECTIVE: To introduce an universal algorithm for kinship index between a baby and a random person with biologic mother reference. METHODS: Based on the formulas of paternity index in trios (PIT), common factors shared in these formulas were deduced following reconstructions of these formulas with the common factors. Universal algorithms for other common kinship indices, such as grandparental index (GI), half sibling index (HSI), avuncular index (AI) and first cousin index (CI1st), were investigated according to avuncular index rule and the coefficient of relationship (r). RESULTS: The common factor shared in the formulas for PI(T) calculation was 1 plus reciprocal of the frequency of the allele with identity by state between the alleged father and the detected baby. Two general formulas for PI(T), GI, AI, HSI and CI1st with biologic mother reference were successfully established with the common factor and r value. CONCLUSION The calculation was simplified with the universal algorithms for common kinship indices between random person and the baby with biologic mother reference and the batch arithmetic operation with the universal algorithms can be easily realized with programming.
Algorithms ; Alleles ; Family ; Female ; Forensic Medicine ; Gene Frequency ; Genotype ; Humans ; Male ; Models, Genetic ; Paternity ; Probability

OBJECTIVE: Research of the hair damage due to perming, combing and stretching can be of important value for forensic hair individual identification. METHODS: The normal human hairs were treated with perming combing and stretching, and the keratins of the damage hair were analysed by using SDS-PAGGE and laser densimeter. RESULTS: Perming, combing and stretching brought about hair damage; The keratins of the damage hair were obviously reduced at the rang of molecular weight of 67,000-43,000 dalton. CONCLUSION The loss of the damage hair keratins were increased with the degree of the hair damage.
Adult ; Female ; Forensic Medicine ; Hair/physiopathology* ; Hot Temperature/adverse effects* ; Humans ; Keratins/metabolism*

OBJECTIVE: The aim was to investigate the polymorphisms of D6S1043 and D12S391 loci among Han population and evaluate their values in paternity testing. MERTHODS: By using fluorescence dye-labeled primers and capillary electrophoresis, the allele frequencies of the two STR loci among 192 unrelated individuals were investigated. RESULTS: Twelve alleles were observed in both D6S1043 and D12S391 loci. The ranges of allele frequencies were from 0.0026 to 0.1719 and from 0.0026 to 0.2292, respectively. The discrimination power of D6S1043 and D12S391 were 0.9656 and 0.9510. The Average exclusion probability in paternity testing for duos were 0.573 and 0.510. The Average exclusion probability in paternity testing for trios were 0.731 and 0.679, respectively. The genotypes frequencies met Hardy-Weinberg equilibrium expectation. CONCLUSION The results show that D6S1043 and D12S391 have high values in forensic paternity testing.
Alleles ; China/ethnology* ; DNA Fingerprinting/methods* ; Electrophoresis, Capillary ; Forensic Medicine/methods* ; Gene Frequency ; Genetics, Population ; Humans ; Paternity ; Probability ; Tandem Repeat Sequences/genetics*