Proteomic Difference Analysis of Whole Blood and Bloodstains / 法医学杂志

OBJECTIVES@#To study the changes of protein levels in peripheral blood after it dried.@*METHODS@#The proteins from whole blood and bloodstains were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and normalized by the label-free quantification (LFQ) method. The differential proteins were analyzed by using R 4.2.1 software, limma and edgeR package. The analysis of biological function, signaling pathway and subcellular localization for the differential proteins was then performed.@*RESULTS@#A total of 623 and 596 proteins were detected in whole blood and bloodstains, respectively, of which 31 were statistically significant in the quantitative results, including 10 up-regulated and 21 down-regulated proteins in bloodstains.@*CONCLUSIONS@#The protein abundances in whole blood and bloodstains are highly correlated, and the variation of protein abundances may be related to the changes of endogenous and structural proteins in cells. The application of proteomics technology can assist the screening and identification of protein biomarkers, thereby introducing new biomarkers for forensic research.

Comparison of MPure-12 Automatic Nucleic Acid Purification and Chelex-100 Method / 法医学杂志

<p><b>OBJECTIVES</b>To explore the forensic application value of MPure-12 automatic nucleic acid purification （MPure-12 Method） for DNA extraction by extracting and typing DNA from bloodstains and various kinds of biological samples with different DNA contents.</p><p><b>METHODS</b>Nine types of biological samples, such as bloodstains, semen stains, and saliva were collected. DNA were extracted using MPure-12 method and Chelex-100 method, followed by PCR amplification and electrophoresis for obtaining STR-profiles.</p><p><b>RESULTS</b>The samples such as hair root, chutty, butt, muscular tissue, saliva stain, bloodstain and semen stain were typed successfully by MPure-12 method. Partial alleles were lacked in the samples of saliva, and the genotyping of contact swabs was unsatisfactory. Additional, all of the bloodstains （20 μL, 15 μL, 10 μL, 5 μL, 1 μL） showed good typing results using Chelex-100 method. But the loss of alleles occurred in 1 μL blood volume by MPure-12 method.</p><p><b>CONCLUSIONS</b>MPure-12 method is suitable for DNA extraction of a certain concentration blood samples．Chelex-100 method may be better for the extraction of trace blood samples．This instrument used in nucleic acid extraction has the advantages of simplicity of operator, rapidity, high extraction efficiency, high rate of reportable STR-profiles and lower man-made pollution.</p>

Forensic Investigation of Goldeneye™ DNA ID 22NC Kit / 法医学杂志

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.

Establishment of miniSTR fluorescent detection system and its forensic application / 法医学杂志

OBJECTIVE@#To establish miniSTR fluorescent detection system with all detected fragments below 150 bp and to enhance the efficiency of detecting the degraded DNA samples.@*METHODS@#All candidate primers were designed by Primer Premier 5 and screened by FastPCR 6.0. The miniSTR multiplex system was established by these selected loci labeling by four fluorescent dye. The parameters of PCR and primer concentrations were subsequently optimized. The electrophoresis was fulfilled under POP4 on 3100-Avant and the typing data was validated by standard DNA 9947A and 007. Fresh blood samples and difficult degraded DNA samples were tested to evaluate the usefulness of the system.@*RESULTS@#All amplicons in the established miniSTR fluorescent detection system (D12ATA63, D2S1776, D1GATA113, D4S2408, D17S974, D20S482, D3S3053, Amelogenin, D6S474, D9S1122) were less than 150bp. The profile showed a balanced peak height without extra stutter by optimal protocol. Allele frequencies showed no deviations from Hardy-Weinberg equilibrium. The system showed accumulated probability of discrimination 0.999 999 983 and accumulated triplet excluding probability of paternity 0.996 8. It could detect corrupt muscle tissue, low copy number DNA samples and human tissues fixed by 40% formaldehyde solution for 12 days.@*CONCLUSION@#The miniSTR fluorescent detection system could be solely used for personal identification of degraded DNA samples or complementally used for paternity tests. And the system could enhance the ability of detecting the trace and degraded DNA.

Forensic validation of goldeneye? DNA ID 26Y system / 法医学杂志

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.

Algorithms of likelihood ratio for discriminating full sibling from half sibling / 法医学杂志

OBJECTIVE@#To derive the formulae for likelihood ratio calculation in discriminating full sibling from half sibling with single-parent participation or without parent participation.@*METHODS@#Null hypothesis and alternative hypothesis were established for discriminating full sibling from half sibling in two circumstances: two children with single-parent and without parent participation. Conditional probabilities of the genetic evidentiary under null and alternative hypotheses were calculated according to the Bayesian theory. The likelihood ratios were established with the conditional probability under alternative hypothesis division that under null hypothesis, followed with simplification. All the formulae were validated in a real case.@*RESULTS@#While mother or fathers' genetic information available in differentiating full sibling from half sibling, 14 different genotype combinations could be shared by the two detected children at a given locus and the likelihood ratio could be calculated with 5 different formulae respectively. While both parents' genetic information unavailable, 11 different genotype combinations could be shared and the likelihood ratio could be calculated with 7 different formulae respectively. It was validated in a real case that the power of the likelihood ratio method developed for discriminating full sibling from half sibling with single-parent participation was higher than that of the ratio of full sibling index over half sibling index.@*CONCLUSION@#The formulae of likelihood ratio developed are useful for discriminating full sibling from half sibling with single-parent participation or without parent participation.

Forensic application of investigator HDplex kit in Han nationality of Eastern China / 法医学杂志

OBJECTIVE@#To investigate the genetic data of 12 autosomal STR loci included in Investigator HDplex kit and to evaluate its forensic application in Han nationality of Eastern China.@*METHODS@#A total of 484 unrelated healthy individuals in Han nationality of Eastern China were investigated with Investigator HDplex kit. Allele frequencies, population genetics parameters and linkage disequilibrium information of the 12 autosomal STR loci were statistically analyzed.@*RESULTS@#No deviations from Hardy-Weinberg equilibrium were detected and all loci were independent form each other within the studied 484 unrelated healthy individuals. DP values of the 12 autosomal STR loci were all above 0.8, and CDP was 0.999 999 999 92. The cumulative probability of paternity exclusion in duo and in trio were 0.999 82 and 0.999 998 6, respectively.@*CONCLUSION@#Investigator HDplex kit with 12 highly polymorphic STR loci in Han nationality of Eastern China could be used effectively for forensic DNA genotyping.

A new modified forked flap and a reverse V shaped flap for secondary correction of bilateral cleft lip nasal deformities / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The columella, nasal tip, lip relationship in the bilateral cleft lip nasal deformity remains a great challenge for plastic surgeon. An esthetically satisfying result is difficult to obtain. A subset of patients with bilateral cleft lip nasal deformity still require columellar lengthening and nasal correction and philtrial construction. This study aimed to provide a new method based on the forked flap to improve the final appearance of these patients.</p><p><b>METHODS</b>A technique to correct this deformity is described. This consists of (1) a newly modified forked flap including the orbicularis oris muscle and nasalis muscle along the whole flap for columellar lengthening, (2) a reverse V shaped flap from the lower portion of the columella and the prolabium for normal size phitrum construction, (3) inserting the vermilion portion of the forked flap and advancing the nasal floor medially and anteriorly to lengthen and maintain the nasal septum side of the columella for proper tip positioning, (4) open rhinoplasty, allowing definitive repositioning of the lower lateral cartilages, (5) reconstruction of the orbicularis orismuscle as required, and (6) the flaring nostril floor advancing medially and constructing the sill.</p><p><b>RESULTS</b>This technique was applied to 15 cases of secondary bilateral cleft lip nasal deformity. All the flaps took without signs of partial necrosis. In all cases, the nasal tip was projected forward with adequate columella elongation, and the height of the prolabium was added with normal size philtrial dimensions.</p><p><b>CONCLUSIONS</b>This method makes maximum use of the tissue containing the scar in the lip and limits tissues in the lower portion of the columella and the prolabium for adequate columella elongation and reconstruction with normal size philtrial dimensions. It is a very reasonable and useful method in correction of secondary bilateral cleft lip nasal deformities.</p>

Establishment of universal algorithms for commonly used kinship indices between two individuals / 法医学杂志

OBJECTIVE@#To establish universal algorithms for commonly used kinship indices between two individuals.@*METHODS@#Based on the formulas of paternity index in duos(PID), full sibling index(FSI), half sibling index (HSI), avuncular index (AI), grandparental index (GI) and first cousin index (CI1st) deduced from ITO method, the common factors, 1 plus reciprocal of the frequency of the allele with identity by state between the two individuals, shared in these formulas were abstracted with induction method, following with reconstruction of these formulas with the common factor and the coefficient of relationship (r).@*RESULTS@#A universal algorithm for PI(D), HSI, AI, GI and CI1st, was developed with the common factor and r value according to the heterozygosity of the two individuals. Meanwhile, a group of two formulas for FSI calculation was also established according to the individuals' heterozygosity.@*CONCLUSION@#The universal algorithms for the 6 types of kinship indices are practical in corresponding kinship determination and the batch arithmetic operation with the universal algorithms can be easily programmed.

SNP typing of formalin-fixed tissue in individual identification / 法医学杂志

OBJECTIVE@#To explore the method for DNA typing in formalin-fixed tissue by detecting SNP markers on X chromosome.@*METHODS@#Genomic DNA was prepared from formalin-fixed tissue. In the event that typing using Sinofiler and MiniFiler kits had failed, 51 SNPs were amplified with multiplex PCR and typed with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). RESULTS Full profiles of X-SNP loci were obtained from the formalin-fixed tissue, while the STR and miniSTR genotyping failed.@*CONCLUSION@#SNP genotyping technique can be used to obtain more information for formalin fixed tissues.

STR profiling of genomic DNA from HE stained tissue sections / 法医学杂志

OBJECTIVE@#To develop a STR analysis method for analyzing DNA from stained tissue sections and to evaluate the capability of this protocol in forensic application.@*METHODS@#Eight kinds of HE stained human tissue, for example heart, liver, lung and intestine, were collected from two autopsy cases. The genomic DNA from those tissues was extracted using a QIAgen kit. DNA quantitation was performed using the TaqMan PCR method. The concentration of DNA isolated was determined based on Ct values. Internal positive controls (IPC) were used to monitor inhibitors. DNA amplifications were performed using Identifiler PCR Amplification kit. PCR products were analyzed on 3100-Avant Genetic Analyzer.@*RESULTS@#The concentrations of DNA obtained from all samples were greater than 1 ng/microL. PCR inhibition was not observed. However, DNA degradation, potentially due to the effect of residual formalin fixative, was observed among tissue samples stored for long periods of time.@*CONCLUSION@#Sufficient amounts of DNA were extracted from HE stained tissue sections. STR profiles were successfully generated. The number of genotype alleles detected decreased as sample storage time increased.

Correction of secondary deformity of unilateral cleft lip with refined anatomic repair in orbicularis oris muscle / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the functional repair of secondary deformity of unilateral cleft lip.</p><p><b>METHODS</b>The nasal branch, nasolabial branch, and labial branch of orbicularis oris muscle were dissected and repositioned precisely to correct the secondary deformity of unilateral cleft lip.</p><p><b>RESULTS</b>96 patients were treated successfully with this method during Jan. 2005 to Oct. 2008. Good cosmetic and functional results were achieved. 85 cases were followed up for 3 months to 5 years with a satisfactory rate of 94.1% (80/85 cases).</p><p><b>CONCLUSIONS</b>The application of refined anatomy and precise reposition in orbicularis oris muscle is important to ensure therapeutic effect in patients with secondary deformity of unilateral cleft lip.</p>

Genotype validation of 12 STR loci by sequencing method / 法医学杂志

OBJECTIVE@#To validate the genotype of 12 STR loci by sequencing method.@*METHODS@#According to the special sequence of 12 STR loci (CSF1PO, FGA, TH01, TPOX, VWA, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 and D21S11), the relative PCR primers were designed. PCR products of control DNA (9947A) and STR mutation samples were sequenced.@*RESULTS@#The sequencing results of both the control DNA (9947A) and STR mutation samples were consistent with their genotyping results.@*CONCLUSION@#The sequencing method developed are accurate and sensitive, can be used for the validation of STR genotyping results.

STR genotyping in unbuffered formalin fixed paraffin embedded tissue / 法医学杂志

OBJECTIVE@#To assess the influential factors of STR genotyping in 10% unbuffered formalin fixed paraffin embedded samples.@*METHODS@#Eight kinds of autopsy samples including heart, brain, liver, spleen, kidney, lung, stomach and intestine tissue from 2 corpse were fixed with 10% unbuffered formalin and embedded with paraffin according to the routine procedure from which the DNA were extracted with three different methods (QIAGEN, IQ and Chelex). STR profile were analyzed with AmpFlSTR Identifiler Kit and capillary electrophoresis on genetic analyzer 3100-Avant. STR profiles of 56 archival paraffin embedded samples from 15 cases were also analyzed with methods as mentioned. These archival samples, including heart, liver, lung and intestine tissue, had been preserved for 1 to 5 years in ambient temperature. Effectiveness of STR genotyping was assessed with the recalling ration of the 15 STR loci composing of the Identifiler Kit.@*RESULTS@#Significant difference of the recalling ration was statistically revealed among the different types of paraffin embedded sample with same preserving period. Moreover, the STR recalling ration was continuously lowering with the prolongation of preserving period in all of the samples. The linear relationship between the STR recalling ratio and the preserving period was showed in lung and heart sample. The STR recalling ration in lung sample was higher than that in the other types of paraffin embedded sample.@*CONCLUSION@#Preserving period, tissue type, extracting method of DNA and the PCR template concentration were the most important influential factors for successfully STR genotyping paraffin embedded samples, which fixed with unbuffered formalin for the same time.

Trilobate technique, a new principal to repair cleft lip / 中华整形外科杂志

<p><b>OBJECTIVE</b>To develop a new method for reparation of cleft lip, and to evoke more colleagues for advance practices and study, in order to determine her indication and contraindication as soon as possible.</p><p><b>METHODS</b>48 cases were included into this study. Trilobate flap were designed in floor of nose and lip area in cleft side, rotate two of the three flaps upwards, respectively to elevate the tip of nose, and to reconstruct the floor of nose. As for the left flap, it was derived transversally to opposing side, sutured with the flap of non-cleft-side.</p><p><b>RESULTS</b>With this technique, less tissue was lost, better vertical lengthening and good formed cuspids-bow was achieved, and the scar was a parallel line being symmetry to the philtrum column opposite. Meanwhile, because the tension was mainly located in the area where there was no mini flaps, the blood supply was good enough, rarely occur any necrosis in the tip of flaps. All cases in this study obtained perfect healing, with good appearance at nostrils and floor of nose.</p><p><b>CONCLUSIONS</b>In use of the method of trilobate flap, we can draw down the peak of the cuspids bow effectually, hence avoid the addition cut in the lower part of the lip, decrease the scar on skin, as well as nice reconstruction of floor of nose, philtrum column and nostril. Because lack of long term study, we evoke more colleagues for cooperation in advance study.</p>

Analysis of genetics and genomics of short tandem repeat loci commonly used in Kinship Testing / 法医学杂志

Since the foundation of FBI laboratory's Combined DNA Index System (CODIS) a decade ago, the 13 CODIS STR loci of the system as well as the recently developed Penta D, Penta E, D2S1338 and D19S433 loci have been widely used by kinship testing laboratories worldwide and have played an important role in the field of Kinship Testing and in construction of criminal database. This article systemically analyzed the characteristics of STR loci information and its genomic information analyzed through search of a variety of database including Webof Knowledge, Elsevier and Internet resources. The up-to-date application of the commonly used STR loci in recent years is also reviewed.

Mutations of short tandem repeat loci in Identifiler system / 法医学杂志

OBJECTIVE@#To explore and analyze the mutations of 15 Short Tandem Repeat (STR) loci using Identifiler system in paternity identification.@*METHODS@#2712 cases of paternity testing were carried out using Identifiler PCR Amplification Kit.@*RESULTS@#Of the 2362 paternity testing cases, mutations of single locus were observed in 51 cases. The mutation loci included D8S1179, D21S11, D7S820, CSF1PO, D3S1358, D13S317, D16S539, D2S1338, D19S433, vWA, D18S51, D5S818 and FGA, with the D21S11 locus having a highest mutation rate (0.369%). Thirty-six of the STR mutations were from paternal source, 7 from maternal source, and the rest (9) were undeterminable. The mutation rates at D21S11 were highest (0.369%).@*CONCLUSION@#Mutations of STR loci are relatively common in human genome. Therefore, retesting of additional relatively stable STR loci with lower mutation rates is necessary when one or two loci exclusions are encountered in paternity testing.

Effect of liposuction on insulin sensitivity and lipid metabolism / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the effect of liposuction on insulin resistance and lipid metabolism.</p><p><b>METHODS</b>The levels of serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and insulin sensitivity were measured pre-and 2-4 months postoperatively in 20 consecutive patients undergoing liposuction.</p><p><b>RESULTS</b>Compared with preoperative, the insulin sensitivity increased significantly, the levels of TC and LDL-C decreased after the liposuction procedure.</p><p><b>CONCLUSIONS</b>Liposuction may improve the insulin resistance and lipid metabolism.</p>

Role of MR imaging on the diagnosis of velopharyngeal insufficiency preoperatively / 中华整形外科杂志

<p><b>OBJECTIVE</b>To assess the feasibility and actuality of MR imaging on diagnosis and evaluation of velopharyngeal insufficiency preoperatively.</p><p><b>METHODS</b>Since 2002, six patients with velopharyngeal insufficiency were examined with MRI using midsagittal, coronal, and axial images, contrast with radiography, to affirm configuration and movements of soft palate, posterior and lateral pharyngeal walls, and velopharyngeal opening for the choices of following surgeries.</p><p><b>RESULTS</b>MRI visualizes directly and measures objectively the shorter soft palate, confined movements of soft palate, lateral or/and posterior pharyngeal wall, and deeper and enlarged velopharyngeal opening, which were coincided with clinical symptoms and affirmed by observations in the following operations, surpassing radiography.</p><p><b>CONCLUSIONS</b>MRI is effective, actual, and uninvasive in imaging and measuring the velopharyngeal insufficiency, thus has a potential role in investigation and planning surgical repairs.</p>

Buccal musculomucosal flap for reconstruction of wide vermilion and orbicularis oils muscle defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the reconstruction of wide vermilion and orbicularis oris muscle defect with satisfactory outcome of aesthetics, sensation, and function.</p><p><b>METHODS</b>The buccal musculomucosal flap based on the anterior buccal branches of the facial artery was used to reconstruct wide defect of vermilion and orbicularis oris muscle on upper or lower lip.</p><p><b>RESULTS</b>7 patients were treated. 5 cases had no postoperative complication. Partial mucosal necrosis on the tip of the flaps happened in 2 cases, but the underlying muscle survived and was re-mucosalized spontaneously. No other complication was observed. The sensation of cold, heat and touch could be detected on the first postoperative day. Electromyographic and electron microscopic studies confirmed innervation of the muscle in the flap.</p><p><b>CONCLUSIONS</b>The buccal musculomucosal flap is a reliable reconstruction option for wide defect of vermilion and orbicularis oris muscle which can' t be reconstructed with conventional method. Satisfactory aesthetic and functional results can be achieved with the buccal musculomucosal flap.</p>