Visual Analysis of the External Characteristics and Research Hotspots of Literatures in International Forensic Science.

ABSTRACT: Objective To analyze the forensic science-related literature included in the Web of Science database in the recent decade through bibliometric methods, to provide reference for relevant research. Methods Literatures were searched in 3 ways： Subject search, Journal search and Institution search. The annual distribution, national （regional） distribution, institution distribution, journal distribution and the research hotspots of the related literatures were analyzed through Thomson Data Analyzer （TDA）, Ucinet, VOSviewer, and so on. Results A total of 49 469 related literatures were included in the recent decade. The number of literatures continued to climb year by year. The top 15 countries （regions） accounted for 78.52% of the total number of published literatures, and China ranked 5th, but ranked 12th in terms of the proportion of high-cited papers; Netherlands, Switzerland, Australia, etc. had high comprehensive influence. The number of countries （regions） that cooperated with China were 129, including the United States, the United Kingdom and Germany. The Institute of Forensic Science of Saint Mary's University, University of Sydney and Netherlands Forensic Institute had high comprehensive influence, and the related literatures were published on 6 357 journals. According to high-frequency co-occurrence network and high-cited papers, brain injury, health policy, assessment scales and models and medical imaging were selected as research hotspots. Conclusion The total number of literatures in forensic science included in international SCI increased significantly, and the influence of China's achievements needs to be greatly enhanced; the research institutions were scattered, and China's research power needs to be continuously condensed; the research hotspots in international fields are extensive, and the international participation of China in top level research needs to be strengthened.

[Association study between candidate genes involved in cell-cell adhesion and non-syndromic cleft lip with or without cleft palate in Chinese population].

OBJECTIVE: To explore the association and gene-environment interaction between single nucleotide polymorphisms (SNPs) involved in cell-cell adhesion and non-syndromic cleft lip with or without cleft palate (NSCL/P) among Chinese population. METHODS: A total of 806 NSCL/P trios were drawn by an international consortium, which conducted a genome-wide association study (GWAS) using a case-parent trio design to investigate genes affecting risks to NSCL/P. The transmission disequilibrium test (TDT) was used to explore the association between cell-cell adhesion genes, including CDH1, CTNNB1, PVRL1, PVRL2, PVRL3, ACTN1, VCL, LEF1, and NSCL/P. Conditional Logistic regression models were used to estimate effects on risk of exposed and unexposed children. Four common maternal exposures including maternal smoking, environmental tobacco smoke, alcohol consumption and multivitamin supplementation during pregnancy were included in this study. RESULTS: A total of 226 SNP markers were tested after quality control in this study. Although 23 SNPs in three genes (CTNNB1, CDH1, ACTN1) showed nominal significant association with NSCL/P in the TDT (P<0.05).There were no significant evidence of linkage and association that remained in the transmission disequilibrium test after Bonferroni correction(P>0.000 2). Tests for gene-environment interaction yielded significant results between rs743127 in ACTN1 and environmental tobacco smoke (P=0.000 1) with an estimated OR (case|G and E)=2.00(95%CI: 1.23-3.26) and OR (case|G no E)=0.59 (95%CI: 0.38-0.90). Among the lower P value results in gene-environment tests, there were no significant results between rs1475034, rs370535, rs2273419 in ACTN1, rs106871 in CTNNB1 and environmental tobacco smoke interaction. There were also no significant results between rs7634000, rs2971366, rs2634553, rs1489032, rs7624812 in PVRL3 and multivitamin supplementation during pregnancy in gene-environment tests(P>0.000 2). CONCLUSION: There is no association between cell-cell adhesion genes, including CDH1, CTNNB1, PVRL1, PVRL2, PVRL3, ACTN1, VCL, LEF1, and NSCL/P when the genes are considered alone. But our results suggest that SNPs in ACTN1 may influence the risk to NSCL/P through gene-environment interaction.

[Analysis of large deletion of phenylalanine hydroxylase gene in Chinese patients with phenylketonuria].

OBJECTIVE: To analyze the types and distribution of large deletion of phenylalanine hydroxylase (PAH) gene in Chinese patients with phenylketonuria (PKU). METHODS: On the basis of 953 PKU patients from Peking Union Medical College and Gansu Province Medical Genetics Center, which were detected by directed sequencing of PAH gene between 2006 and 2014. Multiplex ligation-dependent probe amplification (MLPA) of PAH gene was performed in 43 patients with one or two unknown genotypes. And the deletion breakpoints were characterized by Gap PCR-sequencing. RESULTS: Twenty-four large deletion/duplication alleles were found in 22 patients, accounting for 51.1%(24/47)of the 47 unknown mutations of the 43 patients.There were 6 different large deletions, including Ex1del3758 (n=10), Ex4_5del (n=4), Ex4_7del (n=3), Ex1del5329ins56 (n=3), Ex3del6599ins8 (n=2), and Ex4del (n=1); and 1 duplication was found (Ex12dup, n=1). The most common large deletions in Chinese patients were Ex1del3758 (21.3%), Ex4_5del (8.5%), and Ex4_7del (6.4%). CONCLUSIONS: Large deletion mutations of PAH gene are present in Chinese PKU patients. It's important to detect the large del/dup mutation, and there are different hotspot mutation genotypes in Chinese patients.

Effect and mechanism of dihydroartemisinin on proliferation, metastasis and apoptosis of human osteosarcoma cells.

Osteosarcoma represents an aggressive type of bone malignancy that poses a significant health threat. The objective of the current study was to analyze the effect and mechanism of dihydroartemisinin (DHA) on the proliferation, metastasis and apoptosis of human osteosarcoma cells. A gradient concentration of DHA (15, 25 and 35 µmol.L-1) was used to stimulate the cells, along with control and Dimethyl sulfoxide (DMSO). The phenotypic outcomes were characterized using MTT assay, clone formation assay, Hoechst 33258 staining assay, luciferase reporter plasmid assay, Western blot and wound healing assay. In addition, IBM SPSS Statistics 18.0 software was applied for statistical analysis and all experimental data were expressed as mean ± s.d. Analysis of variance (ANOVA) was applied to compare the differences among multiple groups. Our results demonstrated that DHA inhibited the proliferation and metastasis of osteosarcoma cells and promoted the apoptosis in the cytomorphosis.

[The roles of two HIV self-testing models in promoting HIV-testing among men who have sex with men].

Objective: To evaluate the roles between two different HIV self-testing models in promoting HIV-testing among men who have sex with men (MSM). Methods: This paper focuses on two HIV self-testing service models. The first; is the online self-testing model (HIV self-testing conventional model) with the sexual health promotion network platform. The other one is an innovative HIV self-testing model (secondary distribution model), based on the previous program. The two different self-testing models, including the number of indexes and alters, the positive rate, and the demographics of indexes and alters, are compared. The influence of volunteers with or without leadership on the number of HIV self-test kits distributed or self-use is analyzed through the leadership survey scale. Results: The return rates of HIV self-testing results in the two models are 94.7%(323/341) and 99.2%(1 141/1 150), respectively, within 30 days. The proportion of alters in the secondary distribution is significantly higher (45.9%,281/612) than the conventional HIV self-testing (6.3%,20/318). In the secondary distribution model, the difference between the number of indexes and alters indicators including age, marital status, residence, sex orientation, anal sex with men in the past six months, and HIV test are statistically significant (χ2 test, all P<0.05). The opinion leader of MSM has significantly impacted the promotion of HIV self-testing (P<0.05). Conclusions: Both models can promote HIV self-testing, result return, and HIV positive detection among MSM. In terms of expanding the testing and detection of HIV positive, the secondary distribution mode shows more obvious advantages, which significantly promotes a large number of MSM who have never been tested for HIV to undergo HIV testing. Influential indexes have a significant effect on increasing the HIV testing rate and promoting HIV testing among MSM.

[Cost-effectiveness of HIV self-testing strategy in men who have sex with men].

Objective: To analyze the cost-effectiveness and willingness-to-pay of HIV self-testing (HIVST) strategy and facility-based HIV rapid testing (HIV-RDT) strategy in men who have sex with men (MSM) in Zhuhai, and provide scientific evidence for making health policy. Methods: From the perspective of health service providers, the data of the costs and effectiveness of two HIV testing strategies in MSM in Zhuhai during January-September 2019 were collected, and a decision-tree model of cohort of 10 000 MSM was constructed by using software TreeAge Pro 2019 to measure the cost-effectiveness ratio (CER) and the incremental cost-effectiveness ratio (ICER). One-way and probability sensitivity analysis was performed for the uncertainty of the parameters in the model, and the cost-effectiveness and affordability curve was introduced to estimate the affordability of two strategies. Results: After the mobilization of MSM community-based organization through Internet and social media, 2 303 MSM had HIVST, in whom 33 were HIV positive (1.7%), and 816 MSM received HIV-RDT, in whom 35 were HIV positive (4.3%). The cost for per screening was 60.45 yuan and 240.43 yuan (RMB) respectively, and the cost for per positive screening was 4 218 yuan and 5 606 yuan (RMB) rerspectively. The results of the decision-tree model showed that the mean cost for a MSM using HIVST and using HIV-RDT was 44.67 yuan and 148.42 yuan (RMB) respectively, and the ICER was negative. HIVST strategy was a more cost-effective option when the willing-to-pay was below 6 528 yuan (RMB) for per positive screening, and HIV-RDT strategy was a more cost-effective option when the investment was higher than 6 528 yuan (RMB). Conclusion: HIVST strategy in Zhuhai is a public health project with economic value, and policy makers should strengthen the support to MSM community-based organization to promote the application of HIVST among MSM.

Clinical observations and methods for identifying the existence of cultured epidermal allografts.

Thirty-two burned or plastic surgery patients were grafted with allogeneic cultured epidermis on autograft donor sites. Two techniques, the indirect enzyme conjugated Staphylococcus Protein A assay with monoclonal antibodies against A or B blood group antigens and the polymerase chain reaction to detect a Y chromosome-specific DNA sequence, were employed to identify the presence of cultured epidermal allograft based on different ABO blood grouping or sex between donor and recipient. The methods have the advantage of high sensitivity and specificity in identifying the existence of allogeneic skin cells in grafts. The results indicated that the survival time of cultured epidermal allograft was prolonged up to 35 days. In addition, the intact coverage on some grafting sites may be composed of both host and donor origin cells, after about 3 weeks postgrafting.

Selection of in vitro produced, transgenic embryos by nested PCR for efficient production of transgenic goats.

The production of valuable pharmaceutical proteins using transgenic animals as bioreactors has become one of the goals of biotechnology. However, the efficiency of producing transgenic animals by means of pronuclear microinjection is low. This may be attributed in part to the low integration rate of foreign DNA. Therefore, a large number of recipients are required to produce transgenic animals. We recently developed a transgenic procedure that combined the techniques of goat oocyte in vitro maturation (IVM), in vitro fertilization (IVF), microinjection, preimplantation selection of the transgenic embryos with nested PCR and transferring the transgenic embryos into the recipient goat uterus to produce transgenic goats. Thirty-seven transgenic embryos determined by nested PCR were transferred to thirty-two recipient goats. In the end, four live-born kids were produced. As predicted, all the live kids were transgenic as identified by PCR as well as Southern blot hybridization, The integration rate was 100% (4/4) which was completely in accordance with the results of embryo preimplantation detection. The results showed a significant decrease in the number of recipients required as only 8 recipients (32/4) were needed to obtain one live transgenic goat. We suggest that the transgenic system described herein may provide an improved way to efficiently produce transgenic goats on a large scale.

Amplification of the C-erbB-2(HER-2/neu) proto-oncogene in ovarian carcinomas.

C-erbB-2(HER-2/neu) proto-oncogene is mainly expressed in epithelial tissue and activated due to its amplification. Amplification of the C-erbB-2 proto-oncogene has been associated with poor prognosis in human ovarian cancer. Our study was to examine whether amplification is more frequently observed in ovarian cancer, or it is associated with poor prognosis of human ovarian cancer in China. The DNA of ovarian cancers was extracted and consequently digested with restriction endonuclease EcoRI, electrophoresed in 0.8% agarose gels and blotted onto nitrocellulose filter with Southern transferring method. It was then hybridized with a 32P-labelled C-erbB-2 probe and subsequently underwent autoradiography. The result has shown that the C-erbB-2(HER-2/neu) gene was amplified in 8 of 26 human ovarian cancers (30.8%). The clinical data showed that all of the 8 cases with the amplified C-erbB-2 were in their advanced stage (III-IV). Five of the patients died from 2 to 4 months after operation. These data suggest that amplification of the C-erbB-2 gene may play a role in the pathogenesis of ovarian carcinoma; it is frequently observed in advanced ovarian cancer and is associated with poor prognosis for these patients.

[Endodermal sinus tumor (yolk sac tumor)--report of 52 cases and review of the literature of 1224 cases].

52 cases of endodermal sinus tumors (EST) are reported and 1224 cases from the Chinese and foreign literature are reviewed. This tumor, occurring usually in the gonads and rarely at extragonadal sites, comprised 10-17.1% of the former. It is located on or along the mid-line of the body including the orbit and other organs. In this paper, there are two cases with the tumor located at the orifice of urethra and one case between the pubic bone and the urinary bladder which has never been reported. EST often occurs in people under the age of 35. EST in the gonadal sites is divided into infant-childhood and adult groups. In the former group the prognosis is better than that of the adult group when the tumor is located in the testis as well as those in the vagina. There is sex and age difference for some EST in extragonadal sites. Those in the mediastinum almost always occur in young males. Diagnosis is easily made by pathology and by serum alpha-fetal protein (SAFP). Pathologically it is divided into two types: simple EST and EST element mixed with other germ cell tumors which give the same prognosis. The authors suggest that SAFP be done for any patients with teratomata in order to avoid misdiagnosis. SAFP is not only diagnostic but also prognostic by monitering postoperative course. EST is a highly malignant tumor, rapidly growing and poor in prognosis. Recent reports show that operated patients supplemented by chemotherapy give much better results than by radiotherapy.

[A study of transgenic IFV cattle integrated with human serum albumin gene].

The mammary gland expression vector (pcDNA 3.1-GCALBm) containing the full-length sequence of human serum albumin (hALB) cDNA and intron 1 as well as the goat beta-casien gene promoter and 5' up-stream regulatory sequence was constructed. The vector was micro-injected into bovine IVF eggs. The embryos were in vitro cultured to the late stage of morulae, and then few embryo cells were aspirated for the implantation detection of target gene integration and SRY DNA sequence using nested-PCR. Afterwards, ten integrated embryos were selected to transfer into eight recipients and three were pregnant. The pregnant rate was 37.5%(3/8). However, two were miscarried in mid-trimester but one was pregnant at term to deliver a male transgenic cattle integrated with hALB mini-gene. The transgenic efficiency was 12.5% (1/8).

[Detection of sickle cell gene by analysis of amplified DNA sequences].

This paper describes a technique of DNA amplification in vitro and its application on detection of sickle cell (Hb S) gene. Genomic DNA was microextracted from dried blood specimen of the first patient with sickle cell trait in China. Target DNA sequence was amplified by the polymerase chain reaction (PCR) with the primers beta 1 (5'-ACACAACTGTGTTCACTAGC-3') and beta 2 (5'-CAACTTCATCCACGTTCACC-3') that primed amplification of an 110-base-pair (bp) segment of beta globin gene. The amplified DNA was digested with a restriction endonuclease Mst II, which has a recognition site at codon 6 in the normal beta globin gene, and cleaved the normal amplified beta globin DNA into two fragments of 54bp and 56 bp which was as an overlap band in agarose gel electrophoresis, while the 110bp fragment amplified from DNA of sickle cell mutation remained uncleaved owing to a single base substitution (A----T) at codon 6 in the mutation. DNA amplification method is rapid, sensitive and simple, and does not require radioactive probes. Besides, the PCR amplification can be carried out on the DNA extracted from dried blood samples. So the technique is very useful for gene diagnosis and carrier screening of genetic disease.

[Determination of fetal sex by amplification of Y-chromosome-specific DNA].

This paper describes a rapid and highly sensitive method for determination of fetal sex. Y-chromosome-specific sequences as well as Alu-specific sequences were amplified with polymerase chain reaction (PCR). Then fetal sex was determined by comparison of the two amplified DNA sequences. Polymerase chain reaction can be performed on lysed amniotic fluid cells or chorionic villus samples or dried blood spots on filter paper blot without prior DNA extraction. The analysis of the amplified DNA was performed immediately by agarose gel electrophoresis without DNA hybridization with radioactive probe. By using this method, determination of sex was performed on 3 fetuses at risk for DMD, and on 2 fetuses at risk for hemophilia, prenatal detection was confirmed by examination of the neonates.

[The relation between prognosis and amplification of the c-erB-2 (HER-2/neu) proto-oncogene in ovarian carcinomas].

C-erbB-2 (HER-2/neu) proto-oncogene is mainly expressed in epithelial tissue and activated due to its amplification. Amplification of the C-erbB-2 proto-oncogene is associated with poor prognosis in human ovarian cancer. We examined whether amplification of C-erbB-2 is common in ovarian carcinoma or is associated with poor prognosis. The DNA of ovarian carcinoma was extracted and consequently digested with restriction endonuclease EcoRI, electrophoresed in 0.8% agarose gels and blotted onto nitrocellulose filter with Southern transfering method. It was hybridized with a 32p-labelled C-erbB-2 probe and subsequently underwent autoradiography. It was shown that the C-erbB-2 (HER-2/neu) gene was amplified in 8 of 26 human ovarian carcinomas (30.8%). Clinically the 8 patients with the amplified C-erbB-2 were in their advanced stage (III-IV). Five of the patients died from 2 to 4 months after operation. These findings suggest that amplification of the C-erbB-2 gene may play a role in the pathogenesis of ovarian carcinoma, it is frequently observed in advanced ovarian carcinoma and associated with poor prognosis for these patients.

A new silent mutation found in the Chinese PAH locus and its role in the prenatal diagnosis of phenylketonuria.

A silent mutation or sequence polymorphism. A to T substitution at codon 399 in exon 11 of the PAH gene from a Chinese PKU patient, was found by sequence analysis. The frequencies of this new mutation in normal and abnormal (PKU) genes were 0.005 and 0.09, respectively, based on the analyses of 100 normal individuals and 39 PKU patients using DNA amplification with polymerase chain reaction (PCR) and oligonucleotide hybridization methods. This silent mutation can be used as a "genetic marker" for PKU prenatal diagnosis. Recently, a fetus at risk for PKU, who could not be completely predicted by RFLPs linkage analysis, was prenatally diagnosed with this genetic marker.

Study of the RNA splicing defect in the common Chinese beta-thalassemia gene, IVS-II nt. 654 C-->T by using mRNA/PCR.

With direct sequencing of the amplified cDNA, we analysed the transcript and mRNA splicing defect in a common Chinese beta-thalassemia mutant (IVS-II nt. 654 C-->T). The result shows that this mutant gene would not only produce abnormally processed beta-globin mRNA, but also transcribes a small amount of normally spliced mRNA, hence leading to beta+ thalassemia. The method described herein provides a simple and sensitive approach to the studies of gene expression and molecular defects in genetic diseases at transcriptional level.

Treatment of beta-thalassemia with hydroxyurea (HU)--effects of HU on globin gene expression.

A newly developed method of RT-PCR/competitive PCR for measuring the relative and absolute content of globin mRNAs as well as micro-globin chain biosynthetic assay have been used to study the alterations of globin gene expressions in the patients with beta-thalassemia pre- and post-hydroxyurea (HU) treatment. It was found for the first time that HU had the effect of enhancing beta-globin gene expression in some patients. Two cases with beta-thalassemia who were subjected to HU treatment for over two years showed a marked increase in beta-globin mRNA level and beta-globin chain synthesis, resulting in more effective erythropoiesis and the alleviation of clinical symptoms.

Sexing bovine embryos using PCR amplification of bovine SRY sequence.

This study analyses the bovine SRY DNA sequence by direct sequencing procedure, followed by the designation of the PCR primers specific for bovine SRY. Using PCR amplification of bovine SRY gene, the embryo sex was determined. The results of the embryo sex identification were confirmed after the embryo transfer and pregnancies.

[Research on immune complex in situ type glomerulonephritis treated with mai-luo-tong in rabbits].

Applying c-BSA to duplicate immune complex in situ type glomerulonephritis in rabbits and treating it with Blood Circulation Promoting and Stasis-Removing Drugs Mai-Luo-Tong, the results showed that proteinuria in the treated group was decreased significantly, as compared to the control group (P < 0.01). Under light and electron microscope, although glomerular basement membrane was irregularly thickened and subepithelial dense electron deposits were found in both groups, but histopathologic damage in the treated group less than that of control one. In the treated group micro-thrombus, erythrocytes and platelets aggregation, leukocytes impaction were not seen within glomerular capillary. Also in the treated group mesangial cell proliferation and granulocyte infiltration were decreased significantly (P < 0.01) and there was no apparent glomerular fibrosis in former group.

Aberrant expression of imprinted genes and their regulatory network in cloned cattle.

Domesticated animals cloned by somatic cell nuclear transfer (SCNT) generally have poor developmental competency, with many developmental abnormalities attributed to incomplete reprogramming of the nuclear genome and abnormal expression of genes important for regulation of growth and development. To investigate the molecular mechanism leading to the abnormalities of cloned animals, pathologic and histologic analyses were conducted on seven cloned cattle that were oversized at birth and had cardiac and pulmonary abnormalities. Quantitative reverse transcription (RT)-polymerase chain reaction (PCR) analysis of four imprinted genes IGF2, IGF2R, H19, and GRB10, as well as genes from related regulatory networks, were performed in liver, lung, kidney, and muscle to investigate disruption of expression. Expression of IGFBP2 was not detected in morphologically normal cloned cattle, but was detected in the liver, lung, kidney, and thymus of abnormal calves. Expression levels of IGF1 and imprinted genes IGF2 and H19 were substantially higher in these organs of abnormal cattle. In contrast, expression levels of GRB10, CTSD, and TRPV2 were substantially lower in abnormal cattle. Transcript abundance of IGFBP6 was higher in kidney, but lower in liver and lung. In conclusion, we inferred that altered expression of imprinted and related genes may be closely related to increased birth weight and pathologic changes in abnormal cloned cattle.