Comparative of Forensic DNA Identification Using Cell Lysis Method and Magnetic Beads Method / 法医学杂志

OBJECTIVES@#To compare the effects of cell lysis method and magnetic beads method in forensic DNA identification and to explore these two methods in forensic DNA identification.@*METHODS@#The genome DNA of THP-1 cells in different quantities was extracted by the cell lysis method and magnetic beads method, and the DNA content was quantified by real-time quantitative PCR. The cell lysis method and magnetic beads method were used to type the STR of human blood with different dilution ratios.@*RESULTS@#When the numbers of THP-1 cell were 100, 400 and 800, the DNA content extracted by cell lysis method were (1.219±0.334), (5.081±0.335), (9.332±0.318) ng, respectively; and the DNA content extracted by magnetic beads method were (1.020±0.281), (3.634±0.482), (7.896±0.759) ng, respectively. When the numbers of THP-1 cells were 400 and 800, the DNA content extracted by the cell lysis method was higher than that by the magnetic beads method. The sensitivity of cell lysis method and magnetic beads method was similar in STR typing of human blood at different dilution ratios. Complete STR typing could be obtained at 100, 300 and 500-fold dilutions of blood samples, but could not be detected at 700-fold dilution. STR typing of undiluted human blood could not be detected by cell lysis method.@*CONCLUSIONS@#The cell lysis method is easy to operate and can retain template DNA to the maximum extend. It is expected to be suitable for trace blood evidence tests.

(+)-corynoline Regulates the Proliferation,Stemness and Apoptosis of Triple Negative Breast Cancer Cells / 中国医学科学院学报

Objective To explore the performance and mechanism of(+)-corynoline in treating triple negative breast cancer MDA-MB-436 cells and thus provide an option for the development of drugs against this cancer. Methods The viability,proliferation,apoptosis and migration/invasion of MDA-MB-436 cells treated with(+)-corynoline were detected by CCK-8 assay,colony formation assay,flow cytometry and Transwell assay,respectively.Furthermore,Western blotting was employed to determine the expression of related proteins,and RNA-Seq was performed for the MDA-MB-436 cells treated with(+)-corynoline. Results (+)-corynoline inhibited the proliferation and stemness and promoted the apoptosis of MDA-MB-436 cells.Further,(+)-corynoline may activate the oxidative phosphorylation pathway to play a role in inhibiting triple negative breast cancer. Conclusion (+)-corynoline can inhibit triple negative breast cancer cells,which helps to address the poor efficacy of existing chemotherapeutics and facilitate the development of drugs against this cancer.

Genetic Structure Analysis of Y Chromosome STR and SNP in Population of Changshu / 法医学杂志

Objective To provide a theoretical basis for building a Y chromosome database in specific regions by analyzing the pedigree specific core haplogroup and region specific genetic structure in Changshu. Methods One thousand seven hundred and two samples from unrelated Han male individuals in Changshu were collected. Then 27 Y-STR were genotyped through YfilerTM Plus PCR Amplification Kit, Y-SNP haplogroup of each sample was speculated using Y-Predictor software and some samples were verified by amplification refractory mutation system-polymerase chain reaction （ARMS-PCR）. Results A total of 1 556 haplotypes were found on the 27 Y-STR genetic markers of the 1 702 samples. The haplotype diversity （HD） value was 0.999 827. DYS385 （0.933） had the highest gene diversity （GD） value while DYS438 （0.409） had the lowest. By the Y-Predictor software, all samples were confirmed to be from 162 sub-haplogroups of C, D, N, O, Q and R. Samples were randomly selected to verify the prediction results by the software and the prediction accuracy of Y-Predictor software was as high as 95.74%. Conclusion This study found that 27 Y-STR genetic markers have relatively high polymorphisms in the Changshu population, and have good forensic individual identification and paternity testing ability.

Application prospect of oncolytic virus detection of circulating tumor cell in precision medicine of bladder cancer / 医学研究生学报

Morbidity and mortality rates of bladder cancer (BC) are still rising with a poor prognosis. Therefore, better biomarkers are urgently needed to advance the accurate diagnosis and treatment of BC. The limitations of the different detection techniques of circulating tumor cell (CTC) cause that the CTC as biomarkers of point of view of diagnosis and treatment of BC is not yet clear. This review first compares and analyzes the current CTC detection technology methods, and then reviews the five aspects of screening, diagnosis, staging, curative effect monitoring, prognostic evaluation, and personalized treatment of patients with malignant tumors with oncolytic virus and CTC. The application of oncolytic virus detection CTC in BC was evaluated. The results suggest that oncolytic virus with fluorescent protein combined with fluorescence microscopy and flow cytometer are used to detect CTC. This method can be only used to detect live CTCs, instead of dead CTCs. The CTC count is more accurate, efficient with high sensitivity and specificity. It can also perform phenotypic analysis of CTC and single-cell sequencing. It can be used for screening, diagnosis, and guidance of targeted therapy for bladder cancer, assessing efficacy and judging prognosis which has a very broad clinical application prospect for advancing the accurate diagnosis and treatment of BC.

Application prospect of oncolytic virus detection of circulating tumor cell in precision medicine of bladder cancer / 医学研究生学报

Morbidity and mortality rates of bladder cancer (BC) are still rising with a poor prognosis. Therefore, better biomarkers are urgently needed to advance the accurate diagnosis and treatment of BC. The limitations of the different detection techniques of circulating tumor cell (CTC) cause that the CTC as biomarkers of point of view of diagnosis and treatment of BC is not yet clear. This review first compares and analyzes the current CTC detection technology methods, and then reviews the five aspects of screening, diagnosis, staging, curative effect monitoring, prognostic evaluation, and personalized treatment of patients with malignant tumors with oncolytic virus and CTC. The application of oncolytic virus detection CTC in BC was evaluated. The results suggest that oncolytic virus with fluorescent protein combined with fluorescence microscopy and flow cytometer are used to detect CTC. This method can be only used to detect live CTCs, instead of dead CTCs. The CTC count is more accurate, efficient with high sensitivity and specificity. It can also perform phenotypic analysis of CTC and single-cell sequencing. It can be used for screening, diagnosis, and guidance of targeted therapy for bladder cancer, assessing efficacy and judging prognosis which has a very broad clinical application prospect for advancing the accurate diagnosis and treatment of BC.

Genetic Structure Analysis of Y-Chromosome STR and SNP in Population of Wujiang Area, Suzhou City / 法医学杂志

Objective To analyze the genetic phenotypes of Y-chromosome STR and SNP in Han male population of Wujiang area, Suzhou City and explore the genetic structure of population of Wujiang area for further examination of regional-specific Y-SNP genetic markers ancestor haplogroups. Methods Blood samples of 472 Wujiang area Han males were randomly collected and genotyped by YfilerTM Plus PCR Amplification Kit. The allele frequencies and haplotype frequencies of each locus were obtained using the direct calculation method. Y-SNP haplogroups of each sample were estimated using Y-Predictor software and verified through experiments by amplification refractory mutation system-polymerase chain reaction （ARMS-PCR）. Results A total of 453 haplotypes were found in the 27 Y-STR genetic markers in 472 Han males of Wujiang area. The haplotype diversity （HD） was 0.997 696 93, among which, the highest gene diversity （GD） value was DYF387S1a/b （GD=0.953 1） and the lowest was DYS438 （GD=0.321 8）. Based on genotyping data of 27 Y-STRs and 472 samples, 132 haplogroups from C, D, N, O and Q, etc downstream Y-SNP haplogroups were estimated and then verified through experiments. Conclusion This study is based on Y-chromosome STR haplotypes, and predicts Y-SNP haplogroups by Y-Predictor software, then uses ARMS-PCR to verify. Y-SNP genetic markers were introduced to achieve precise analysis of the genetic structure of male families in population of three towns in Wujiang area.

Advance in Mendelsohn Maneuver for Dysphagia (review) / 中国康复理论与实践

Dysphagia can lead to multiple complications, increasing mortality and recurrence, and affecting the quality of life. Mendelsohn maneuver is a kind of physical therapy to improve the safety and effectiveness of swallowing. This paper introduced the origin of Mendelsohn maneuver, its physiological mechanism and clinical application. The potential risks and shortcomings of Mendelsohn maneuver were discussed, and a new solution was put forward.

Design of electrode fixation and positioning cap for transcranial electrical stimulation / 医疗卫生装备

Objective To design a new type of rehabilitation device to execute electrode fixation and precision positioning during transcranial electrical stimulation.Methods The device was composed of a positioning cap,a soft rubber keel scaffold, an inlaid nut,a unionbolt,an electrical stimulation shim and etc.The electrode was put into the shim at the target site,and fixation and positioning were realized by regulating the depth of the bolt into the slot to change the attachment between the electrode fixing cushion and cranial skin,and then electrical stimulation therapy could be carried out reliably and accurately. Results The device gained advantages over the traditional means in fixation time in one time, dropping-off rate, patient satisfaction and easy operation.Conclusion This device behaves well in simple structure,easy manufacture,low cost,fixation and large applicability to kinds of population and electrical stimulators,and enhances the reliability of transcranial electrical stimulation therapy to some extent.

Genetic polymorphisms of 15 X-STR loci in Shandong Han population / 法医学杂志

OBJECTIVE@#To investigate the genetic polymorphisms of 15 X-STR loci in Shandong Han population in order to establish the forensic application database.@*METHODS@#The multi-PCR primers of these loci were designed by Primer Premier 5.0 software and labeled by 4 fluoresceins (FAM, VIC, NED and TET). The developed multi-PCR was used to investigate 15 X-STR loci (DXS10011, DXS101, GATA 165B12, DXS6795, DXS6800, DXS6801, DXS6803, DXS7132, DXS7133, DXS7423, DXS7424, DXS8377, DXS8378, DXS9898 and HPRTB) selected from the X chromosome of 481 unrelated individuals (295 females and 186 males) in Shandong Han population.@*RESULTS@#Among the 15 X-STR loci, GATA 165B12, DXS6800, DXS6803, DXS7133 and DXS7423 showed moderate polymorphisms, while the rest 10 X-STR loci showed high polymorphisms (PIC > 0.5 and H > 0.5). No shared haplotype was detected among the males in Shandong Han population.@*CONCLUSION@#The developed multi-PCR system with fluorescence detection provides an effective way to establish X-STR loci database of population genetics in Shandong Han population and shows its forensic application.

Differences of DNA methylation profiles in monozygotic twins' blood samples / 法医学杂志

OBJECTIVE@#To evaluate the potential usefulness of DNA methylation in individual discrimination of monozygotic twins by investigating the differences of DNA methylation profiles in monozygotic twins' blood samples.@*METHODS@#Blood samples from 22 pairs of monozygotic twins were obtained with informed consent. Genomic DNA extracts were bisulfite treated followed by detection with Infinium HumanMethylation27 BeadChip Assays(Illumina, USA). Epigenetic distances between each pair of monozygotic twins and each pair of unrelated individuals of same gender were calculated with Euclidean distance algorithms. Distribution of epigenetic distance in monozygotic twin group was statistically compared with that in unrelated individuals.@*RESULTS@#Difference of epigenetic distance between male and female pairs was not statistically significant in unrelated individual group or in monozygotic twin group (P = 0.0695 and 0.4825, respectively). Epigenetic distance of monozygotic twins was significantly lower than that of unrelated individual pair of same gender (Median: 6.02 vs 7.20, P = 0.0002). However, all the epigenetic distance in monozygotic twin group or in unrelated individuals were significantly higher than 4.00 (P < 0.000 1).@*CONCLUSION@#DNA methylation profiles of monozygotic twin's blood samples were significantly different with each other, which was similar to that in unrelated individuals of same gender. These results indicated that DNA methylation was a useful biomarker in individual discrimination of monozygotic twins.

Genetic screening of mutation hot-spots for nonsyndromic hearing loss in southern Jiangsu province with SNaPshot / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the mutation frequency in 7 mutation hot-spots of deafness gene in southern Jiangsu province and verify the performance of the SNaPshot technology platform, designed for genetic screening of non-syndromic hearing loss (NSHL) in Chinese.</p><p><b>METHODS</b>One hundred and twenty-five NSHL patients were enrolled. Amplification of 235delC, 299-300delAT in GJB2 gene, IVS7-2A>G, 2168 A>G in SLC26A4 gene, and 1555A>G, 7445 A>G and 3243 A>G in mitochondrial DNA (mtDNA) was performed using multiplex polymerase chain reaction (PCR) technology. Afterwards, the sequence-specific probe interrogated each locus and labeled it at the 3' end using fluorescent dideoxynucleotide chemistry by the SNaPshot Multiplex Kit, the resulting products were then separated electrophoretically in ABI PRISM R 3130 Genetic Analyzer and analyzed in the presence of a fifth-dye-labeled size standard. Finally, the genotyping results were verified by direct sequencing or PCR-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>(1) The total mutation frequency for the 7 mutation hot-spots was 53.6%. The mutation frequency of 235delC was 24.0%, 299-300delAT was 5.6% in the GJB2 gene, IVS7-2A>G was 15.2%, 2168A>G was 3.2% in the SLC26A4 gene. The mutation frequency of 1555A>G and 7445 A>G in mtDNA was 4.8% and 0.8% respectively. The mutation 3243 A>G was not detected. (2) The SNaPshot results were consistent with that from direct sequencing or PCR-RFLP, and the specificity and sensitivity of detection were 100%.</p><p><b>CONCLUSION</b>(1) More than half of the patients with deafness in southern Jiangsu province carry the mutations of the seven hot-spots. (2) The genetic screening technology platform based on SNaPshot can detect 7 mutations in one reaction, and is efficient and suitable for clinical practice.</p>

Expression of TRPM and TRPV channel family mRNA in rat spermatogenic cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of transient receptor potential melastatin (TRPM) and transient receptor potential vanilloid (TRPV) channel family genes in rat spermatogenic cells.</p><p><b>METHODS</b>Rat spermatogenic cells were isolated by a mechanical procedure and the total RNA was extracted using TRIzol reagent. TRPM and TRPV channel family genes were amplified by RT-PCR and the presence of the target genes was detected by agarose gel electrophoresis. The relative gene expression levels were measured by real-time quantitative RT-PCR.</p><p><b>RESULTS</b>TRPV5, TRPM3, TRPM4 and TRPM7 mRNAs were expressed in rat spermatogenic cells, but TRPV1, TRPV2, TRPV3, TRPV4, TRPV6, TRPM1, TRPM2, TRPM5, TRPM6, TRPM7 and TRPM8 mRNAs were not detected. The relative expressions of TRPM and TRPV mRNA were determined by quantitative real-time RT-PCR. TRPM7 expression was the highest among all the TRPM subtypes in rat spermatogenic cells, at a level equivalent to (0.0430-/+0.0034)% of beta-actin expression. TRPM3 and TRPM4 were also highly expressed, but their expression levels were only approximately 56% and 63% of that of TRPM7, respectively. For the TRPV subfamily, only TRPV5 mRNA was abundantly expressed at the level of (0.0157-/+0.0029)% relative to that of beta-actin.</p><p><b>CONCLUSION</b>TRPV5, TRPM3, TRPM4 and TRPM7 mRNAs were coexpressed in spermatogenic cells in rats, among which TRPM4 and TRPM7 mRNA were expressed at high levels. TRPM4 and TRPM7 channels may be involved in the regulation of growth, differentiation and maturation of rat spermatogenic cells and are associated with the generation of the sperms.</p>

Dose and long-term effect of hIGF-1 injection for erectile dysfunction in aged rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the best dose and the long-term effect of the human insulin-like growth factor-1 (hIGF-1) gene injection into the penis of aged rats.</p><p><b>METHODS</b>Included in this study were 10 young (4 months old) and 40 aged (24 months old) Sprague-Dawley male rats, the latter equally divided into a PBS control and a 10 microg, a 100 microg and a 1 000 microg hIGF-1 injection group. Electrical stimulation was conducted 4 and 8 weeks after hIGF-1 injection into the penile corpus cavernous of the rats to detect the intracavernous pressure (ICP) and mean arterial pressure (MAP). Dose - and time -associated therapeutic results were analyzed and the mRNA expression of hIGF-1 determined by RT - PCR.</p><p><b>RESULTS</b>ICP, MAP and total ICP were significant decreased by electrical stimulation in the aged rats as compared with the young ones (P < 0.05), statistically increased in the three hIGF-1 dose groups in comparison with the PBS controls (P < 0.05), and showed no obvious difference between the young rats and the latter two dose groups at 4 and 8 weeks. Although less obvious effect was achieved in the 10 microg group than in the young rats, the therapeutic result was still of significance. The mRNA expression of the hIGF-1 gene was confirmed in all the hIGF-1 treated rats.</p><p><b>CONCLUSION</b>The hIGF-1 therapy can improve erectile function in aged rats, 100 microg suffices for effective erection and the effect may last at least 8 weeks for a single dose.</p>

Sonographic patterns of intussusception with relationship to pathologic change,reducibility and ischemia / 中华超声影像学杂志

Objective To investigate sonographic patterns of intussusception with relationship to pathologic change,reducibility and ischemia. Methods Twenty-one intussusceptions were surgically induced in rabbits,and in vitro ultrasonography was compared with the corresponding pathologic changes. Ultrasonography findings in 25 cases of pediatric intussusception confirmed by means of air enema examination or surgery were analyzed. Results The results of animal experiment showed that:①when no ischemia of intestine loop occurred,axial images of intussusception showed a "target-like" mass,if ischmia of intestine loop appeared,it demonstrated as a "doughnut-like" mass; ②bowel unviability and reducibility seemed clearly related to the thickness of hypoechoic external rim of the "doughnut" mass and blood flow on color Doppler flow images (CDFI) in intussusceptum. When the flow on CDFI in intussusception was absent,the possibility of bowel necrosis was increased( P