Objective To investigate the allele frequencies of eight short tandem repeats(STR) loci:TH01,FES, D19S400,D7S820,D16S539,D20S161,D3S1545 and D5S818 in Han population in Henan province.Methods DNA was extracted with phenol-chloroform from EDTA-blood samples of the unrelated individuals in Henan province and amplified with PCR technique. The PCR product was analyzed with the undenatured PAGE vertical electrophoresis and silver-stain.Results The authors got the frequencies of the eight loci. The heterozygosities of the eight loci are 0.66, 0.67, 0.80, 0.76, 0.79, 0.79, 0.78 and 0.78; the discrimination powers are 0.83, 0.83, 0.94, 0.91, 0.93, 0.93, 0.92 and 0.92.Conclusion The heterozygosities of the eight loci are high and the frequencies are in good agreement with Hardy-Weinberg equilibrium, so the eight loci can be used in idividual identification testing.

Objective To investigate the distribution differences of Porphyromonas gingivalis(Pg) FimA genotypes between periodontitis patients with and without type 2 diabetes for a better understanding of the relationship between diabetes and periodontitis.Methods Questionnaires and detailed periodontal examinations(6 sites per tooth) were conducted in 80 subjects with moderate-severe chronic periodontitis in the Department of Periodontology, Peking University School and Hospital of Stomatology.There were 40 type 2 diabetic patients and 40 systemicly healthy patients enrolled respectively.The periodontal parameters including plaque index(PLI), probing depth(PD), bleeding index(BI), attachment loss(AL) by 6 sites per tooth and numbers of missing teeth were also recorded.Pooled subgingival plaque samples using pocket method with Whatman No3 filterpaper were collectedat each of the 6 sites from one incisor and one molar.Pg and its FimA genotype distributions were investigated using DNA extrctedfrom plaque samples by PCR.Results Diabetic patients had a significantly higher score of PLI[2.35(0.58) vs 1.64(0.76), P＜0.05] , while rest of periodontal indexes observed(PD, BI and AL) did not differ significantly between diabetic patients and systemicly healthy controls(P＞0.05).The detection rate of Pg did not show statistically significant difference between the two groups(50％ vs 60％, P＞0.05).However, the proportion of FimA Ⅱ was significantly higher in diabetic group than systemicly healthy group(80％ vs 42％, P＜0.05).Conclusions Type 2 diabetic patients were prone to be infected by highly virulent strains of Pg: FimA Ⅱ Pg.

BACKGROUND:In the initial stage of multiple sclerosis,central immune cells activate and release a large number of inflammatory factors,causing white matter demyelination and even involving gray matter neurons.The equilibrium of differentiation between different subsets of CD4+ T cells plays an important role in the progression of experimental autoimmune encephalomyelitis.The previous results of the research group showed that the active ingredient astragalus glycoprotein in astragalus can regulate the immune response in experimental autoimmune encephalomyelitis mice,and whether it has a regulatory effect on the differentiation of T cell subsets has not been determined. OBJECTIVE:To explore the therapeutic effects and immune regulatory mechanisms of astragaloside IV on experimental autoimmune encephalomyelitis mice. METHODS:Female C57BL/6 mice were divided into the normal control group,experimental autoimmune encephalomyelitis disease model group,and astragaloside IV treatment group(n=8 per group).Myelin oligodendrocyte glycoprotein peptides 35-55 were used for experimental autoimmune encephalomyelitis model induction in the last two groups.On day 10 to 28 after immunization,the astragaloside IV treatment group was treated with 40 mg/kg per day astragaloside IV intragastrically.Body weight and clinical scores of mice in each group were recorded from the immunization day to the 28th day.On the 28th day after immunization,the mouse spinal cord was taken and made into frozen sections for hematoxylin-eosin staining and Lux fast blue staining to observe pathological changes in the spinal cord.Percentage of splenic T cell subsets was detected using flow cytometry.Western blot assay was used to determine the protein expression of interferon-γ,interleukin-17 and interleukin-6 in the spinal cord.Levels of interferon-γ,interleukin-17,interleukin-6 and interleukin-4 in supernatants of cultured splenocytes were determined by ELISA. RESULTS AND CONCLUSION:(1)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could reduce the degree of weight loss in experimental autoimmune encephalomyelitis mice(P<0.05),ameliorate clinical symptoms(P<0.05),inhibit the infiltration of inflammatory cells and alleviate myelin loss(P<0.01,P<0.05).(2)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could inhibit the proportion of CD4+T cell subsets expressing interferon-γ(P<0.001)and interleukin-17(P<0.001),but increase percentages of CD4+ interleukin-10+(P<0.001)and CD4+ transforming growth factor-β+(P<0.01)T cell subsets.(3)Astragaloside IV could inhibit the expression of interferon-γ(P<0.05,P<0.01),interleukin-17(P<0.05,P<0.05),and interleukin-6(P<0.05,P<0.05)in the spinal cord and spleen,and up-regulate the expression of interleukin-4(P<0.01)in spleen.(4)These findings confirm that astragaloside IV alleviates clinical symptoms in experimental autoimmune encephalomyelitis mice,which may be related to regulating the splenic T cell subsets,therefore,inhibiting the infiltration of inflammatory cells into the center and reducing the demyelination.

Objective:To provide experimental evidence for genetic counseling and prenatal diagnosis by analyzing the clinical characteristics, screening and identification of the function of suspicious variants in a X-1inked spondyloepiphyseal dysplasia tarda (SEDT) family.Methods:The family members' medical history, general physical examination, femur, spine X-ray examination were collected. Peripheral blood samples of the family members were collected and DNA was extracted from these samples. Sequencing clinical whole exons of proband DNA by targeted gene high-throughput sequencing method, then analysis sequencing data. The suspicious mutation was confirmed in pedigree members by PCR and Sanger sequencing. Reverse transcription polymerase chain reaction (RT-PCR) experiments of total RNA from blood lymphocytes were performed. The amplification of exons 3 and 4 of the pathogenic gene were amplified and identified by agarose gel. The expression of the pathogenic gene was also detected.Results:Three affected males of the family were diagnosed with SEDT according to their clinical and radiological features. A nonsense mutation in the transport protein particle complex subunit 2 ( TRAPPC2) gene NM_001011658: c.91A>T (p.K31*) was found in the proband using whole exome sequencing. This variation was also detected in his cousin, but not in non-phenotypic members of the family. The RT-PCR result for amplification of exon 3 and 4 of peripheral blood lymphocytes was the same as those of normal controls, indicating that the mutation did not affect the splicing of transcripts. qPCR results showed that the transcriptional expression of TRAPPC2 in patients was significantly lower than that in family normal controls and normal people controls. Conclusion:Identification of the novel nonsense mutation (c.91A>T) in the SEDT family enables early patients screening, carrier detection, genetic counseling, prenatal diagnosis, and clinical prevention and treatment. The detailed genotype/phenotype descriptions contribute to the SEDT mutation spectrum. The study of the function of TRAPPC2 mutation will help to further elucidate the role of sedlin in cartilage.

Objective:To analyze clinical characteristics of and causative genes in two families with dystrophic epidermolysis bullosa, and to reveal the pathogenesis of the disease and mechanisms underlying phenotypic differences between patients.Methods:DNA was extracted from peripheral blood samples of members from two families with dystrophic epidermolysis bullosa, and subjected to high-throughput sequencing and Sanger sequencing.Results:The clinical manifestations of the 2 probands in the 2 families were consistent with the diagnosis of dystrophic epidermolysis bullosa, and the symptoms of the proband in family 1 were more serious than those of other patients in the family. Genetic testing showed that all patients in family 1 carried a mutation c.6082G>C (p.G2028R) in the COL7A1 gene, and the proband and her phenotypically normal mother and uncle also carried a splice-site mutation c.7068+2 (IVS91) T>G in the COL7A1 gene, both of which were first reported. The proband in family 2 carried the mutations c.6081_6082 ins C (p.G2028Rfs*71) and c.1892G>A (p.W631X, first reported) in the COL7A1 gene, which were inherited from her father and mother, respectively.Conclusion:The two pathogenic mutations may be the molecular mechanism underlying the severe clinical phenotype in the proband in family 1; the first reported mutations enriched the mutation spectrum of the COL7A1 gene.

The causes of mental disorders are complex, and early recognition and early intervention are recognized as effective way to avoid irreversible brain damage over time. The existing computer-aided recognition methods mostly focus on multimodal data fusion, ignoring the asynchronous acquisition problem of multimodal data. For this reason, this paper proposes a framework of mental disorder recognition based on visibility graph (VG) to solve the problem of asynchronous data acquisition. First, time series electroencephalograms (EEG) data are mapped to spatial visibility graph. Then, an improved auto regressive model is used to accurately calculate the temporal EEG data features, and reasonably select the spatial metric features by analyzing the spatiotemporal mapping relationship. Finally, on the basis of spatiotemporal information complementarity, different contribution coefficients are assigned to each spatiotemporal feature and to explore the maximum potential of feature so as to make decisions. The results of controlled experiments show that the method in this paper can effectively improve the recognition accuracy of mental disorders. Taking Alzheimer's disease and depression as examples, the highest recognition rates are 93.73% and 90.35%, respectively. In summary, the results of this paper provide an effective computer-aided tool for rapid clinical diagnosis of mental disorders.
Humans ; Mental Disorders/diagnosis* ; Alzheimer Disease/diagnosis* ; Brain Injuries ; Electroencephalography ; Recognition, Psychology

Identification of genetic variants via high-throughput sequencing (HTS) technologies has been essential for both fundamental and clinical studies. However, to what extent the genome sequence composition affects variant calling remains unclear. In this study, we identified 63,897 multi-copy sequences (MCSs) with a minimum length of 300 bp, each of which occurs at least twice in the human genome. The 151,749 genomic loci (multi-copy regions, or MCRs) harboring these MCSs account for 1.98％of the genome and are distributed unevenly across chromosomes. MCRs containing the same MCS tend to be located on the same chromosome. Gene Ontology (GO) anal-yses revealed that 3800 genes whose UTRs or exons overlap with MCRs are enriched for Golgi-related cellular component terms and various enzymatic activities in the GO biological function cat-egory. MCRs are also enriched for loci that are sensitive to neocarzinostatin-induced double-strand breaks. Moreover, genetic variants discovered by genome-wide association studies and recorded indbSNP are significantly underrepresented in MCRs. Using simulated HTS datasets, we show that false variant discovery rates are significantly higher in MCRs than in other genomic regions. These results suggest that extra caution must be taken when identifying genetic variants in the MCRs via HTS technologies.

Objective:To analyze the influences of leukocytes on improving blood glucose control in patients with type 2 diabetes mellitus (T2DM) and periodontitis after periodontal mechanical therapy.Methods:Thirty-five patients visiting Peking University Third Hospital from March 2011 to August 2012, as well as thirty-four patients visiting Peking University School and Hospital of Stomatology from March 2011 to August 2012 and December 2016 to December 2018 were selected in this research. These subjects were non-smokers, and with moderate to severe chronic periodontitis and T2DM. The full set of periodontal examinations including probing depth (PD), attachment loss (AL), bleeding index (BI) and plaque index (PLI) were conducted. Besides, counts of white blood cells (WBC), parameters of glucose and lipids metabolites such as fasting blood glucose (FBG), glycosylated hemoglobin (HbA 1c), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) in serum were examined before treatment. Then, oral hygiene instruction, scaling and root planing (SRP) were carried out. Three months after SRP, the baseline examinations were repeated in all patients. According to the baseline leukocyte counts, the patients were divided into subgroups: low WBC group (WBC<6.19×10 9/L) and high WBC group (WBC≥6.19×10 9/L). Paired t-test for comparison of changes after treatment, analysis of co-variance for comparing the intervention effects between subgroups, and multifactor Logistic regression analysis were performed. Results:Three months after SRP, all periodontal indexes were significantly improved in both groups. Leukocyte counts decreased significantly in high WBC group (6.89±1.53 vs. 7.64±1.51, P=0.008). In high WBC group, HbA 1c (7.18±1.09 vs. 7.67±1.35, P=0.001) and LDL (2.67±0.85 vs. 3.28±0.76, P=0.042) decreased significantly, while there were no such differences in low WBC group. Influence of leukocyte level on HbA 1c ( OR=0.12, P=0.038) and LDL ( OR=0.15, P=0.001) improvement was statistically significant. Hierarchical analysis showed such improvement notably perform in female [HbA 1c ( OR=0.30, P=0.021), LDL ( OR=0.34, P=0.001)] and severe periodontitis group [HbA 1c ( OR=0.15, P=0.025), LDL ( OR=0.24, P=0.017)]. Through interaction test, female and leukocyte counts at baseline had relative excess risk affecting the effect of periodontal intervention on HbA 1c ( P=0.036) and LDL ( P=0.005). Conclusions:SRP could significantly improve the blood glucose and lipid control in patients who had T2DM and chronic periodontitis with relative higher leukocytes level. Female patients with severe periodontitis showed more obviously effects.

The E3 ligase HERC4 is overexpressed in human breast cancer and its expression levels correlated with the prognosis of breast cancer patients. However, the roles of HERC4 in mammary tumorigenesis remain unclear. Here we demonstrate that the knockdown of HERC4 in human breast cancer cells dramatically suppressed their proliferation, survival, migration, and tumor growth in vivo, while the overexpression of HERC4 promoted their aggressive tumorigenic activities. HERC4 is a new E3 ligase for the tumor suppressor LATS1 and destabilizes LATS1 by promoting the ubiquitination of LATS1. miRNA-136-5p and miRNA-1285-5p, expression of which is decreased in human breast cancers and is inversely correlated with the prognosis of breast cancer patients, are directly involved in suppressing the expression of HERC4. In summary, we discover a miRNA-HERC4-LATS1 pathway that plays important roles in the pathogenesis of breast cancer and represents new therapeutic targets for human breast cancer.

OBJECTIVES: To determine the potential molecular mechanisms underlying the therapeutic effect of curcumin on hepatocellular carcinoma (HCC) by network pharmacology and experimental in vitro validation. METHODS: The predictive targets of curcumin or HCC were collected from several databases. the identified overlapping targets were crossed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) platform. Two of the candidate pathways were selected to conduct an experimental verification. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium (MTT) assay was used to determine the effect of curcumin on the viability of HepG2 and LO2 cells. The apoptosis and autophagy of HepG2 cells were respectively detected by flow cytometry and transmission electron microscopy. Besides, western blot and real-time polymerase chain reaction (PCR) were employed to verify the p53 apoptotic pathway and adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK) autophagy pathway. HepG2 cells were pretreated with pifithrin-‍α (PFT-‍α) and GSK690693 for further investigation. RESULTS: The 167 pathways analyzed by KEGG included apoptosis, autophagy, p53, and AMPK pathways. The GO enrichment analysis demonstrated that curcumin was involved in cellular response to drug, regulation of apoptotic pathway, and so on. The in vitro experiments also confirmed that curcumin can inhibit the growth of HepG2 cells by promoting the apoptosis of p53 pathway and autophagy through the AMPK pathway. Furthermore, the protein and messenger RNA (mRNA) of the two pathways were downregulated in the inhibitor-pretreated group compared with the experimental group. The damage-regulated autophagy modulator (DRAM) in the PFT-‍α-pretreated group was downregulated, and p62 in the GSK690693-pretreated group was upregulated. CONCLUSIONS Curcumin can treat HCC through the p53 apoptotic pathway and the AMPK/Unc-51-like kinase 1 (ULK1) autophagy pathway, in which the mutual transformation of autophagy and apoptosis may occur through DRAM and p62.
AMP-Activated Protein Kinases/pharmacology* ; Apoptosis ; Carcinoma, Hepatocellular/pathology* ; Curcumin/pharmacology* ; Humans ; Liver Neoplasms/pathology* ; Network Pharmacology ; Tumor Suppressor Protein p53/metabolism*