ObjectiveTo investigate the characteristics of mitochondrial translational initiation factor 2 （MTIF2） gene methylation and its association with the development and progression of hepatocellular carcinoma （HCC）. MethodsMethSurv and EWAS Data Hub were used to perform the standardized analysis and the cluster analysis of MTIF2 methylation samples， including survival curve analysis， methylation signature analysis， the association of tumor signaling pathways， and a comparative analysis based on pan-cancer database. The independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Cox proportional hazards model was used to perform the univariate and multivariate survival analyses of methylation level at the CpG site. The Kaplan-Meier method was used to investigate the survival differences between the patients with low methylation level and those with high methylation level， and the Log-likelihood ratio method was used for survival difference analysis. ResultsGlobal clustering of MTIF2 methylation showed that there was no significant difference in MTIF2 gene methylation level between different races， ethnicities， BMI levels， and ages. The Kaplan-Meier survival curve analysis showed that the patients with N-Shore hypermethylation of the MTIF2 gene had a significantly better prognosis than those with hypomethylation （hazard ratio ［HR］=0.492， P<0.001）， while there was no significant difference in survival rate between the patients with different CpG island and S-Shore methylation levels （P>0.05）. The methylation profile of the MTIF2 gene based on different ages， sexes， BMI levels， races， ethnicities， and clinical stages showed that the N-Shore and CpG island methylation levels of the MTIF2 gene decreased with the increase in age， and the Caucasian population had significantly lower N-Shore methylation levels of the MTIF2 gene than the Asian population （P<0.05）； the patients with clinical stage Ⅳ had significantly lower N-Shore and CpG island methylation levels of the MTIF2 gene than those with stage Ⅰ/Ⅱ （P<0.05）. Clinical validation showed that the patients with stage Ⅲ/Ⅳ HCC had a significantly lower methylation level of the MTIF2 gene than those with stage Ⅰ/Ⅱ HCC and the normal population （P<0.05）. ConclusionN-Shore hypomethylation of the MTIF2 gene is a risk factor for the development and progression of HCC.

Dipsaci Radix is a commonly used Chinese herbal medicine in China, with triterpenoid saponins as the main active components. β-Amyrin synthase, a member of the oxidosqualene cyclase superfamily, plays a crucial role in the biosynthesis of oleanane-type triterpenoid saponins. Asperosaponin Ⅵ is an oleanane-type triterpenoid saponin. To explore the β-amyrin synthase genes involved in the biosynthesis of asperosaponin Ⅵ in Dipsacus asper, this study screened the candidate genes from the transcriptome data of D. asper. Two β-amyrin synthase genes, Da OSC1 and Da OSC2, were identified by phylogenetic analysis and correlation analysis. The coding sequences of Da OSC1 and Da OSC2 were 2 286 bp and 2 295 bp in length, encoding 761 and 764 amino acids,respectively. Multiple sequence alignments showed that Da OSC1 and Da OSC2 had three conserved motifs( DCTAE, QW, and MWCYCR) unique to the oxidosqualene cyclase family. Real-time quantitative PCR results showed that Da OSC1 and Da OSC2 had the highest expression levels in the roots. Compared with normal growth conditions, the low-temperature treatment significantly upregulated the expression of Da OSC1 and Da OSC2. Agrobacterium-mediated transient expression of Da OSC1 and Da OSC2 in Nicotiana benthamiana resulted in the production of β-amyrin, which suggested that Da OSC1 and Da OSC2 were able to catalyze the synthesis of β-amyrin. This study clarified the catalytic functions of two β-amyrin synthases in D. asper, analyzed their expression patterns in different tissue and at low temperatures. The findings provide a foundation for further studying the biosynthetic pathway and regulatory mechanism of asperosaponin Ⅵ in D. asper.
Intramolecular Transferases/chemistry* ; Phylogeny ; Plant Proteins/chemistry* ; Gene Expression Regulation, Plant ; Dipsacaceae/classification* ; Saponins/metabolism* ; Oleanolic Acid/metabolism*

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

ObjectiveTo observe the effects of Hedysarum polysaccharides（HPS）on the signaling pathways of B-cell lymphoma 2 （Bcl-2）， cysteinyl aspartate-specific protease 3 （Caspase-3）， and Bcl-2-associated X protein （Bax） in Schwann cells（SCs）cultured in high glucose，and explore the possible mechanism of HPS against diabetic peripheral neuropathy（DPN）. MethodFour SD suckling mice aged 5-7 days were randomly divided into a normal group，a high-glucose group，an HPS + high-glucose group，and an α-lipoic acid（α-LA）+ high-glucose group. SCs were extracted from the sciatic nerve and cultured in a 37 ℃，5% CO₂ incubator. After the cells reached 80% confluence，Cell Counting Kit-8（CCK-8）was used to screen the experimental concentrations suitable for high glucose，HPS， and α-LA interventions. Western blot and Real-time polymerase chain reaction （Real-time PCR）were used to detect the protein and mRNA expression of Bcl-2，Bax，and Caspase-3. The apoptosis rate of SCs was detected by flow cytometry using Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI）. ResultAs revealed by Western blot and real-time PCR，compared with the normal group，the high-glucose group showed reduced protein and mRNA expression of Bcl-2 and increased protein and mRNA expression of Bax and Caspase-3（P<0.01）. Compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed increased protein and mRNA expression of Bcl-2 and decreased protein and mRNA expression of Bax and Caspase-3（P<0.01）. As displayed by the results of flow cytometry using Annexin V/PI， compared with the normal group，the high-glucose group showed increased apoptosis rate；compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed reduced apoptosis rate（P<0.01）. ConclusionHPS can alleviate the apoptotic response of SCs，and its mechanism may be related to the inhibition of the activation of the Bcl-2/Caspase-3 signaling pathway.

Objective:To establish the fingerprint of Baoyuantang substance benchmark， and to analyze and identify the common peaks. Method:A total of 15 batches of Baoyuantang substance benchmark were prepared， ultra performance liquid chromatography-diode array detector method （UPLC-PDA） was used to establish the fingerprint of the substance benchmark， and the methodology was developed. The chromatographic conditions were as follows：ACQUITY UPLC BEH Shield C₁₈ column （2.1 mm×100 mm， 1.7 μm）， mobile phase of 0.05% formic acid solution （A） and 0.05% formic acid acetonitrile solution （ B） for gradient elution （0-0.5 min， 5%-19%B； 0.5-6 min， 19%B； 6-10 min， 19%-27%B； 10-20 min， 27%-45%B； 20-20.1 min， 45%-95%B； 20.1-23 min， 95%B）， the flow rate of 0.4 mL·min^-1， the column temperature of 30 ℃， the detection wavelength at 203 nm and 260 nm， and the injection volume of 2 μL. Similarity evaluation system of traditional Chinese medicine fingerprint （2012 edition） was used to establish the fingerprint and generate the control fingerprint. The chemical constituents of Baoyuantang substance benchmark were identified by comparison of standard substances and UPLC-electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） with full information tandem mass spectrometry （MS^E） and scanning range of m/z 50-1 200. Result:The similarities of 15 batches of Baoyuantang substance benchmark were above 0.90 by comparing with the control fingerprint. There were 37 common peaks， 22 of which were identified through UPLC-ESI-MS/MS， including liquiritin， violanthin， ginsenoside Rg₁， ginsenoside Rb₁， ginsenoside Re and so on. These components were all from Astragali Radix， Ginseng Radix et Rhizoma， Zingiberis Rhizoma Recens and Glycyrrhizae Radix et Rhizoma. Conclusion:This method is accurate， stable and reliable， which will basically reflect the overall chemical composition characteristics of Baoyuantang， and it provides experimental basis for development of the granules of this famous classical formulas.

Although DNA 5-hydroxymethylcytosine (5hmC) is recognized as an important epige-netic mark in cancer, its precise role in lymph node metastasis remains elusive. In this study, we investigated how 5hmC associates with lymph node metastasis in breast cancer. Accompanying with high expression of TET1 and TET2 proteins, large numbers of genes in the metastasis-positive pri-mary tumors exhibit higher 5hmC levels than those in the metastasis-negative primary tumors. In contrast, the TET protein expression and DNA 5hmC decrease significantly within the metastatic lesions in the lymph nodes compared to those in their matched primary tumors. Through genome-wide analysis of 8 sets of primary tumors, we identified 100 high-confidence metastasis-associated 5hmC signatures, and it is found that increased levels of DNA 5hmC and gene expression of MAP7D1 associate with high risk of lymph node metastasis. Furthermore, we demonstrate that MAP7D1, regulated by TET1, promotes tumor growth and metastasis. In conclusion, the dynamic 5hmC profiles during lymph node metastasis suggest a link between DNA 5hmC and lymph node metastasis. Meanwhile, the role of MAP7D1 in breast cancer progression suggests that the metastasis-associated 5hmC signatures are potential biomarkers to predict the risk for lymph node metastasis, which may serve as diagnostic and therapeutic targets for metastatic breast cancer.

Major depressive disorder (MDD) has a high disability rate and a high risk of suicide. Due to the limited efficacy of traditional antidepressants, researchers have actively developed animal models with different mechanisms to meet the clinical needs. The principle and preparation methods, phenotypic characteristics, model validity (simulating clinical MDD) and their advantages and disadvantages of different models have been reviewed in this article. It is helpful for researchers to explore the pathogenesis of MDD and to select appropriate animal models in the development of new anti-MDD drugs.

Objective: To investigate the awareness of pubertal knowledge and its influencing factors among primary school students in Chinese rural areas, and to provide evidences for improving awareness level and popularizing scientific, reasonable, and suitable sex education. Methods: Sixteen primary schools consisting of rural registered students in Guizhou and Anhui Province were selected by multi-stage cluster sampling method, and a questionnaire survey for 2 506 senior students (grade five and six) was conducted to explore the awareness of pubertal knowledge and its influencing factors among primary school students in Chinese rural areas. Results: The total awareness rate of pubertal knowledge was 62.7% among primary school students in Guizhou and Anhui rural areas, and the awareness rates were relatively low for the questions about the difference between male and female pubertal development and understanding of sexual attitude; female students, only-child students and grade six students got a bit better awareness of pubertal knowledge than male students, non-only-child students and grade five students respectively; grade[OR=0.72(0.61-0.86)], peer acceptance [OR=0.70(0.53-0.96)], family intimacy [OR=0.75(0.63-0.88)] had effect on awareness of pubertal knowledge. Conclusion The awareness of pubertal knowledge among primary school students in Guizhou and Anhui rural areas need further improvement, especially for male students, non-only-child students and grade five students in the questions about the difference between male and female pubertal development and understanding of sexual attitude. It is necessary to improve the awareness of pubertal knowledge by enhancing peer acceptance, promoting family intimacy, and popularizing scientific, reasonable, and suitable sex education.

Objective: To understand the incidence of school violence and its related factors among senior students in rural primary schools in Guizhou and Anhui provinces in 2016. Methods: From September to November 2016, one county was randomly selected from 47 impoverished counties of Guizhou and one county was randomly selected from 18 impoverished counties of Anhui Province by using multi-stage cluster sampling method. A total of 4 schools were randomly selected from primary schools which had more than 70% students with rural resident registration in each selected county, and 3 classes were randomly selected from fifth and sixth grades of each selected school. All students in each selected class were included in this study. A total of 2 521 students were included, and 2 506 valid questionnaires were collected with an effective response rate about 99.4%. Self-designed questionnaires were used to collect data on individual information, family information, and the incidence of school violence with related factors. Multivariable logistic regression was conducted to analyze related factors of school violence. Results: The incidence of violence in rural primary schools was 50.4% (1 264/2 506) in 2016. The number of students who had physical assault, fought or been bullied was 682 (27.2%), 689 (27.5%) and 764 (30.5%), respectively. The results of multivariable logistic regression showed that the OR (95%CI) values of incidence of school violence for male students was 2.13(1.78-2.54) compared with female students; and the OR(95%CI) values of incidence of school violence for students with high peer acceptance, high family intimacy, more physical activity and mother who had education background of junior high school, senior high school, technical secondary school and junior college, bachelor′s degree or above were 0.39 (0.26-0.57), 0.47 (0.40-0.57), 0.64 (0.52-0.78), 0.79 (0.66-0.96), and 0.65 (0.46-0.92) compared with those with low peer acceptance, low family intimacy, less physical activity and mother who had primary education and below. Conclusion The incidence of violence in rural primary schools is not optimistic. Gender, peer acceptance, family intimacy, physical activity and mother education are related to school violence.

To understand the incidence of school violence and its related factors among senior students in rural primary schools in Guizhou and Anhui provinces in 2016. Methods From September to November 2016, one county was randomly selected from 47 impoverished counties of Guizhou and one county was randomly selected from 18 impoverished counties of Anhui Province by using multi?stage cluster sampling method. A total of 4 schools were randomly selected from primary schools which had more than 70% students with rural resident registration in each selected county, and 3 classes were randomly selected from fifth and sixth grades of each selected school. All students in each selected class were included in this study. A total of 2 521 students were included, and 2 506 valid questionnaires were collected with an effective response rate about 99.4%. Self?designed questionnaires were used to collect data on individual information, family information, and the incidence of school violence with related factors. Multivariable logistic regression was conducted to analyze related factors of school violence. Results The incidence of violence in rural primary schools was 50.4% (1 264/2 506) in 2016. The number of students who had physical assault, fought or been bullied was 682 (27.2%), 689 (27.5%) and 764 (30.5%), respectively. The results of multivariable logistic regression showed that the OR (95%CI) values of incidence of school violence for male students was 2.13(1.78-2.54) compared with female students; and the OR (95%CI) values of incidence of school violence for students with high peer acceptance, high family intimacy, more physical activity and mother who had education background of junior high school, senior high school, technical secondary school and junior college, bachelor′s degree or above were 0.39 (0.26-0.57), 0.47 (0.40-0.57), 0.64 (0.52-0.78), 0.79 (0.66-0.96), and 0.65 (0.46-0.92) compared with those with low peer acceptance, low family intimacy, less physical activity and mother who had primary education and below. Conclusion The incidence of violence in rural primary schools is not optimistic. Gender, peer acceptance, family intimacy, physical activity and mother education are related to school violence.