By analysing the 300 cases of paternity test in our lab in the recent year, we have demonstrated that mutation rates in STR loci are very high. The experimental steps include extracting DNA by chelex 100, amplification of DNA, polyacrylamide gel electrophoresis, silver staining, DNA typing by standard samples. The STR mutations in 11 cases among 300 paternity test cases were found, induding D11S554 locus6 cases, D19S253 locus2 cases, SE33 locuslcase, D12S391 locus 1 case, D13S631 locus 1 case. The results indicated that high mutation rates in STR loci should be considered when STR genotyping are applied to paternity test.

Objective To evaluate the cause which leads to the allelic dropouts at D8S1179 locus while performing paternity testing with the AmpFlSTR Profiler Plus kit. Methods A singleplex amplification system for D8S1179 locus (GenBank Accession No. G08710) was used to verify the typing results by using the AmpFlSTR?Profiler plus kit. Dropout alleles were then sequenced. Results G to A transition was identified at the position of the 147th base of the GenBank sequence. The frequency of the G to A transition among the Chinese population was 0.50 X 10"2 (10 out of 2013 unrelated individuals). Conclusion The G to A transition may be located at the binding site of one of the primers of the AmpFlSTR?Profiler Plus kit. It is suggested that the G to A transition might be the cause of the allelic dropout at the D8S1179 locus.

Objective To explore the diagnostic value of serum creatine kinase isoenzyme MB (CM‐MB) and cardiac troponin I (cTnI) for myocardial injury in children with hand‐foot‐mouth disease (HFMD) .Methods A total of 80 children with HFMD (HFMD group) and 50 healthy children (control group) were enrolled from July 2012 to June 2013 .Serum levels of CK‐MB and cTnI were compared between the two groups .Results Serum levels of CK‐MB and cTnI were (38 .10 ± 19 .50)U/L and (0 .08 ± 0 .02)μg/L in HFMD group ,which were higher than control group (P<0 .05) .In HFMD group ,the positive rate of CK‐MB was 56 .3% ,higher than the 33 .8% of cTnI (P< 0 .05) .After therapy ,serum levels of CK‐MB and cTnI were both significantly de‐creased (P<0 .05) .Conclusion Combined detection of serum CK‐MB and cTnI might be with important significance for the early diagnosis of myocardial injury in children with HFMD .

BACKGROUND:Macropore morphology of a composite scaffold prepared by the three-dimensional printing technique is of great importance in determining the physicochemical and biological properties of tissue engineering scaffolds.OBJECTIVE:To fabricate strontium-containing mesoporous (Sr-MBG) bioactive glass (PCL) scaffolds by the three-dimensional printing technique, and to explore the effect of these scaffolds on MC3T3-E1 proliferation and osteogenic differentiation, thereby to find out the optimal macropore morphology.METHODS: Sr-MBG/PCL composite scaffolds were fabricated by the three-dimensional printing technique. The angles between fibrous latitudes and longitudes were set to 45°, 60° and 90°. Then the proliferation and alkaline phosphatase activity of MC3T3-E1 cells on the scaffolds were tested.RESULTS AND CONCLUSION: Cell counting kit-8 results showed that MC3T3-E1 cells could proliferate on all the three kinds of scaffolds. The proliferation rate of MC3T3-E1 cells on the 45° Sr-MBG/PCL scaffolds was just slightly higher than that on the 60° and 90° Sr-MBG/PCL scaffolds at days 1 and 4 (P > 0.05), but there was a significant increase at day 7 (P < 0.05). The 45° Sr-MBG/PCL scaffolds exhibited a significant increase in alkaline phosphatase activity of MC3T3-E1 cells compared to the 60° and 90° Sr-MBG/PCL scaffolds at day 14 (P < 0.05), while there was no significant difference among three groups at day 21 (P > 0.05). These results indicate that the 45° Sr-MBG/PCL scaffold is more suitable to promote the proliferation and osteogenic differentiation of the MC3T3 cells than the 60° and 90° Sr-MBG/PCL scaffolds.

Objective The aim of this study was to investigate mutations of 41 STR loci. Methods 4546 bloodstain samples were typed from 1932 father–mother–child trios by using AGCU_21+1, AGCU_EX22 and GlobalFiler_ExpressTM amplification Kit. Calculate the mutation rates of STR loci. Results 154 mutations were identified at 32 of the 41 loci. The average mutation rate was 1.0×10-3per locus(95%CI: 0.8~1.1×10-3), and the mutations of SE33 was highest. 152(98.7%) mutation events were one-step mutation, 2(1.3%) events were two-steps. The mutation events occurred in 150 father–mother–child triplets. The mutations in 146(97.3%) triplets occurred at single locus, 8 mutations were observed at two loci in 4(2.7%) triplets simultaneously. 104 paternal and 22 maternal mutations could be determined under 79212 paternal and maternal allelictransfers. The ratio of paternal versus maternal mutations was 4.7:1, and 28 unassigned mutations were observed. Conclusion STR mutation are common in paternity testing, and we should pay more attention to it.

Objective To investigate the effect of Candida albicans and its components on the expression level of human beta-defensin-2 mRNA (HBD-2) in keratinocytes in vitro. Methods Different components of Candida albicans were isolated by lyticase, repeated freezing and thawing, sonication, and centrifugation. The keratinocytes and HaCaT cell lines were co-cultured with Candida albicans and its cellular components for 24 h. The expression level of HBD-2 mRNA was detected by reverse-transcription polymerase chain reaction (RT-PCR). Results Low expression level of HBD-2 mRNA in the unstimulated keratinocytes and HaCaT cells was detected. The HBD-2 mRNA expression levels in the keratinocytes stimulated by Candida albicans, the extract of its cell wall, and pure mannan were significantly increased (P 0.05). Conclusions Candida albicans, the extract of cell wall of Candida albicans, and commercial mannan can increase the expression level of HBD-2 mRNA in keratinocytes.

[Objective] To learn about the genetic diversity,we studied the five X-chromosomal STR (X-STR) loci in Guangdong Han Nationality Groups.[Methods] The five Loci (DXS6803,DXS981,DXS6809,DXS6789,and DXS7132) were amplified in a pentaplex PCR reaction.PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer,with GeneMapper ID 3.1 Analysis Software.[Results] A total of 363 individuals (181 unrelated male and 182 unrelated female) from Guangdong Han population were tested,54 alleles were observed for these loci.Polymorphism information content is 0.6935 ～ 0.8177.Power of discrimination in females was 0.8976 ～ 0.9562.Mean exclusion chance for X-STR in standard trios with daughters was 0.7805 ～ 0.8467.[Conclusion] The five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing,particularly for difficult paternity deficiency cases.

BACKGROUND: Three-dimensional (3D) printing technique has showed unparalleled advantages in the field of tissue engineering scaffold preparation because of its outstanding merits of convenience, efficiency, controllability and ability to construct complex shapes.OBJECTIVE: To fabricate Fe-containing mesoporous calcium-silicate (MCS) /poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) composite scaffolds using the 3D printing technique and to test the characterization and cellular biocompatibility of the composite scaffolds.METHODS: Four groups of Fe-containing MCS/PHBHHx composite scaffolds were fabricated using 3D printing technique. The molar percentage of Fe in these four groups was 0%, 5%, 10%, 15%, respectively and they were marked as 0Fe-MCS/PHBHHx, 5Fe-MCS/PHBHHx, 10Fe-MCS/PHBHHx and 15Fe-MCS/PHBHHx. The scanning electron microscopy was used to observe the microstructure of the scaffolds after being soaked in the simulated body fluid.Osteoblast cell lines MC3T3-E1 were seeded on these four groups of scaffolds as well. Cell counting kit-8 method was adopted to test the cell proliferation at 1, 3, 7 days of culture. Intracellular alkaline phosphatase activity was tested at 7 and 14 days of culture.RESULTS AND CONCLUSION: (1) Compared with the scaffolds with no soaking process, spherical particles were formed on the scaffolds because of mineralization after soaking 3 days in the simulated body fluid. (2) At 1 day of culture,there was no difference in cell proliferation among the four groups. At 3 days of culture, the proliferation rate of the 15Fe-MCS/PHBHHx scaffold was remarkably higher than that of the rest three groups (P < 0.05). At 7 days of culture,the proliferation rate was significantly higher in the 10Fe-MCS/PHBHH and 15Fe-MCS/PHBHHx scaffolds than the 0Fe-MCS/PHBHH scaffold (P < 0.05), as well as significantly higher in the 15Fe-MCS/PHBHHx scaffold than the 10Fe-MCS/PHBHH scaffold (P < 0.05). (3) At 7 days of culture, no difference in alkaline phosphatase activity could be found among these four groups of scaffolds; however, at 14 days, the 5Fe-MCS/PHBHHx, 10Fe-MCS/PHBHHx and 15Fe-MCS/PHBHHx scaffolds exhibited an enhanced alkaline phosphatase activity compared with the 0Fe-MCS/PHBHHx scaffold. Meanwhile, the 15Fe-MCS/PHBHHx showed the highest alkaline phosphatase activity.These findings indicate that the MCS/PHBHH scaffolds containing Fe could promote the proliferation and osteogenic differentiation of the MC3T3-E1 cells.

Objective To investigate the duodenum absorptive character of monosialotetrahexosylganglioside sodium (GM1) in rats.Methods The contents of phenolsulfonphthalein (as indicators) and GM1 were determined with ultraviolet-visible (UV) method and high performance liquid chromatography (HPLC) method in rats with in situ cycle intestinal perfusion model.Results The ratio of duodenum absorption of GM1 was 10％ in 2 h after cycle and 22％ in 6 h after cycle,respectively.The Ka was (0.030± 0.012)h,and absorption t1/2 was (25.50 ± 8.56)h in 8 h after cycle.Conclusions GM1 is absorption in rat duodenum,and the accumulate absorption of GM1 is almost linearly related to the cycle time.The absorption dynamics of GM1 may be first-order kinetic process.

OBJECTIVETo explore the clinical features and mutation of TGFBI gene in a Chinese pedigree affected with lattice corneal dystrophy (LCD).

METHODSGenomic DNA was extracted from 35 members including 11 patients from the pedigree. The 17 exons and splicing region of introns of the TGFBI gene were amplified by PCR. The products were directly sequenced and compared with GenBank database to identify potential mutation. Bioinformatic analysis was carried out to predict the effect of mutation on proteins.

RESULTSA heterozygous mutation (p.R124C) was found in exon 4 of the TGFBI gene in all patients from the pedigree but not among unaffected members. The mode of inheritance of corneal dystrophy in this pedigree was identified as autosomal dominant. Bioinformatics analysis predicted that the p.R124C mutation may be functionally deleterious. The phenotype of corneal dystrophy in the pedigree was determined to be LCD I type.

CONCLUSIONThe p.R124C mutation of the TGFBI gene probably underlies the pathogenesis of LCD in this Chinese pedigree. Genetic testing can facilitate proper diagnosis of this type of corneal dystrophy.