Objective:To evaluate the role of the sodium leak channel (NALCN) in the hippocampal dentate gyrus (DG) in the social behavior of mice.Methods:Thirty-nine male wild-type C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were used in this study. Three mice were sacrificed to verify the expression and co-expression of NALCN with neuronal nuclear antigen (NeuN) in the hippocampal DG using the immunofluorescent staining. The remaining 36 mice were divided into 2 groups ( n=18 each) by the random number table method: control group (group C) and NALCN gene knockdown group (group KO). NALCN-shRNA virus was injected in group KO, and scrambled-shRNA virus was injected in group C. The three box social test and open field test were performed at 3 weeks after the virus injection. Mice were sacrificed under anesthesia after the behavioral test, hippocampal tissues were collected, and the injection location of the virus was verified with a fluorescence microscope, and the NALCN protein and mRNA expression in the hippocampal DG was detected by Western blot and real-time polymerase chain reaction, respectively. Results:NALCN and NeuN co-expressed a lot on the same neuron in the hippocampal DG of mice, indicating that NALCN was widely expressed on the neurons in the hippocampal DG. Compared with group C, the expression of NALCN and mRNA in the hippocampal DG was significantly down-regulated, and the social novelty preference disappeared ( P<0.05), and no significant change was found in the social ability and each parameter in the open field test in group KO ( P>0.05). Conclusions:NALCN in the hippocampal DG is involved in the regulation of social memory in mice, and the down-regulated expression of NALCN can lead to the loss of social novelty preference in mice.

Objective A knowledge graph construction pipeline of traditional Chinese medicine(TCM)diagnosis and treatment was designed and applied,aiming at the automatic construction of the"disease-symptom-pathogenesis-and medicine"knowledge graph based on the medical records of famous TCM physicians,to analyze,organize and present medical records efficiently.Methods Firstly,The entity extraction method of medical records combining Deep Learning and Regular Expression was designed to extract disease,symptom,pathogenesis,and TCM entities from unstructured medical records automatically;secondly,entity relationships were defined and the correlations between entities were calculated using HAN method,and then the"entity-relation-entity"triplets were built;the graph database Neo4j and Gephi were used for knowledge storage and visual display separately;Finally,the application was verified in the Medical records of lung cancer treated by the old famous TCM physicians.Results The precision,Recall and F1 of the knowledge extraction model for medical records entities extraction are 88.49%,90.02%and 89.25%,respectively,and each index is better than the comparison methods.A total of 1077 triples are extracted through entity correlation calculation,and the knowledge graph is successfully constructed.It can reflect the relationship between 'disease-symptom-pathogenesis-medicine' in the treatment of lung cancer by the famous specialists of TCM.Conclusion The method in this paper can effectively solve the extraction,organization and expression of clinical medical records of famous TCM physicians,and realize the automatic construction process from the text of medical records to the knowledge graph.Relevant research ideas and methods proposed in this paper could provide a reference for the construction of the diagnosis and treatment knowledge graph of famous TCM physicians based on medical records.

Klebsiella pneumoniae(K.pneumoniae)is an important pathogen that can cause severe hospital-and community-acquired infections.To systematically investigate its methylation features,we determined the whole-genome sequences of 14 K.pneumoniae strains covering varying serotypes,multilocus sequence types,clonal groups,viscosity/virulence,and drug resistance.Their methy-lomes were further characterized using Pacific Biosciences single-molecule real-time and bisulfite technologies.We identified 15 methylation motifs[13 N6-methyladenine(6mA)and two 5-methylcytosine(5mC)motifs],among which eight were novel.Their corresponding DNA methyl-transferases were also validated.Additionally,we analyzed the genomic distribution of GATC and CCWGG methylation motifs shared by all strains,and identified differential distribution pat-terns of some hemi-/un-methylated GATC motifs,which tend to be located within intergenic regions(IGRs).Specifically,we characterized the in vivo methylation kinetics at single-base resolu-tion on a genome-wide scale by simulating the dynamic processes of replication-mediated passive demethylation and MTase-catalyzed re-methylation.The slow methylation of the GATC motifs in the replication origin(oriC)regions and IGRs implicates the epigenetic regulation of replication initiation and transcription.Our findings illustrate the first comprehensive dynamic methylome map of K.pneumoniae at single-base resolution,and provide a useful reference to better understand epigenetic regulation in this and other bacterial species.

Objective:To report four male COL4A5 mutation mosaicism patients with X-linked Alport syndrome, and to provide evidence for diagnosis, genetic counseling, and reproduction in the respective families and improve our knowledge of mosaicism in Alport syndrome. Methods:Suspected male mosaic patients for COL4A5 who met the following criteria: clinical diagnosis of Alport syndrome, harbored COL4A5 mutations detected using next generation sequencing or Sanger sequencing, heterozygosity for the mutant and normal COL4A5 alleles in the DNA demonstrated by Sanger sequencing, registered in the on-line registry of hereditary kidney diseases, and admitted to Peking University First Hospital during the period of April 2018 to April 2019 were enrolled. Clinical data and karyotypes were retrospectively analyzed. Genetic DNA isolated from multiple tissues was analyzed for COL4A5 gene mutations by using PCR and Sanger sequencing. Related literatures published in PubMed, CNKI and Wanfang databases were reviewed. Results:Four COL4A5 somatic and germline mosaic male patients with Alport syndrome were included in the study. Patient 1 was characterized by hematuria and proteinuria. His karyotype of peripheral blood was normal. COL4A5 c.3455-1G>A mosaicism was detected in multiple tissues (peripheral blood, saliva and urine). Patient 2 presented with hematuria and microalbuminuria. His karyotype of peripheral blood was normal. COL4A5 c.4994+1G>A mosaicism was detected in multiple tissues (peripheral blood, saliva and skin fibroblasts). Patients 3 showed hematuria without proteinuria. COL4A5 c.3535G>A mosaicism was found in genomic DNA of peripheral blood and hair. Laboratory and physical examinations of patient 4 showed hematuria and normal renal function, without proteinuria, megasoma or small testes. COL4A5 c.3106G>A mosaicism was detected in genomic DNA of skin fibroblasts. Although without karyotype analysis due to unavailable specimens, 47,XXY or 46,XY/47,XXY mosaicism was not considered according to the reproductive history and lack of clinical manifestations of megasoma and small testes in patients 3 and 4. Renal disease in 8 published male cases with mosaicism for COL4A5 was affected by mutant allelic fractions and genotype. Conclusions:Compared with hemizygous males with X-linked Alport syndrome, the renal phenotype of mosaic males was milder, and associated with mutant allelic fractions and mutation type.

Objective:To unravel the CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas from EWSR1 rearrangement-negative undifferentiated round cell sarcomas in the bone and soft tissues.Methods:Twenty-eight cases of EWSR1 rearrangement-negative undifferentiated round cell sarcomas of bone and soft tissues, tested for CIC rearrangement and BCOR rearrangement by fluorescence in situ hybridization and related immunostaining were analyzed, and some of the BCOR rearrangement cases were verified by reverse transcription-polymerase chain reaction.Results:Five of 28 (17.9%) tested cases were positive for CIC rearrangement and six (21.4%) for BCOR rearrangement. Histopathologically, CIC rearrangement sarcomas comprised nodular aggregates of round to polygonal cells, containing hyperchromatic nuclei, prominent nucleoli and moderate cytoplasm, with focal variable necrosis and myxoid stroma. BCOR-CCNB3 sarcomas mostly comprised diffusely arranged, round to oval to short spindly cells with angulated nuclei, vesicular chromatin, inconspicuous nucleoli and interspersed vessels. Immunohistochemically, five of six BCOR-CCNB3 sarcomas showed CCNB3 immunostaining, which could be helpful for diagnosis . Two patients with CIC rearrangement sarcoma died of the diseases in seven months and twenty-two months. One patient with BCOR-CCNB3 sarcoma died of the diseases in forty-six months. Conclusions:Overall, 39.3% of the EWSR1 rearrangement-negative undifferentiated round cell sarcomas are CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas. Molecular testing is helpful for diagnosis.

Objective: To investigate the clinical features, diagnosis, occurrence sequence and clonal origin of chronic lymphocytic leukemia complicated with multiple myeloma. Methods: The diagnosis and treatment of one patient with multiple myeloma and chronic lymphocytic leukemia who was admitted to the First Hospital of Jilin University in May 2018 was retrospectively analyzed, and the related literatures were reviewed. Results: This patient began with lumbosacral pain, and he was diagnosed as chronic lymphocytic leukemia complicated with multiple myeloma after bone marrow aspiration, flow cytometry, and blood and urine immunofixation electrophoresis. It is recommended that Rd (lenalidomide + dexamethasone) or MPV (melphalan + prednisone + bortezomib) regimen, but the patient did not receive chemotherapy and died of infectious diarrhea 1 month later. Conclusions The occurrence of multiple myeloma and chronic lymphoblastic leukemia may originate from the same clone or different new clone. It is very rare that multiple myeloma and chronic lymphoblastic leukemia can co-occur. Therapeutic options tend to be more aggressive multiple myeloma-based regimen.

Objective To evaluate the effect of the treatment with low-concentration hydrogen peroxide (H2O2) on the adhesive function of and autophagy in human melanocytes,and to screen long noncoding RNAs (lncRNAs) related to autophagy.Methods Melanocytes were isolated from foreskins of healthy males after circumcision,and subjected to cultivation.Melanocytes at exponential growth phase were divided into 3 groups:control group receiving no treatment,H2O2 group treated with 400 μ mol/L H2O2,and H2O2 + NAC group pretreated with 4 mmol/L NAC for 2 hours followed by the treatment with 400 μmol/L H2O2.After 5-day treatment,immunofluorescence study was performed to determine the expression of Ecadherin,microtubule-associated protein 1 light chain 3 (LC3)and p62,and Western blot analysis to determine the expression of autophagy-related protein LC3 and p62.Cell structures and autophagosomes were observed by transmission electron microscopy,and autophagy-related lncRNAs were screened using gene chip technology.Statistical analysis was done with Graphpad Prism 6 software using one-way analysis of variance for comparison among groups,and Tukey's test for multiple comparisons.Results Under the confocal microscopy,the H2O2 group showed significantly decreased fluorescence intensity of E-cadherin and LC3 in the melanocytes and decreased number of autophagosomes in melanocytes,but significantly increased fluorescence intensity of p62 compared with the control group and H2O2 + NAC group.Western blot analysis showed that the LC3-Ⅱ/LC3-Ⅰ ratio in the melanocytes was significantly lower in the H2O2 group (0.604 ± 0.012) than in the control group (1.200 ± 0.081,q =7.718,P ＜ 0.01) and H2O2 + NAC group (1.017 ± 0.062,q =5.076,P ＜ 0.05),while the p62/β-actin ratio in the melanocytes was significantly higher in the H2O2 group (0.881 ± 0.079) than in the control group (0.456 ± 0.121,q =4.847,P ＜ 0.05) and H2O2 + NAC group (0.492 ± 0.049,q =4.439,P ＜ 0.05).There were no significant differences in the LC3-Ⅱ/LC3-Ⅰ ratio or p62/β-actin ratio between the H2O2 + NAC group and control group (P ＞ 0.05).Gene chip technology showed that 18 autophagy-related lncRNAs were associated with premature senescence of melanocytes and differentially expressed in the H2O2 group compared with the control group,and the autophagy-related lncRNA NONHSAT190308.1 (＞ 10-fold increase) was screened out.Conclusion Lowconcentration H2O2 can decrease the expression of E-cadherin and the level of autophagy in melanocytes,and can up-regulate the expression of autophagy-associated lncRNA NONHSAT190308.1.

Objective? To investigate the hope level of elderly patients undergoing total knee arthroplasty (THA) and analyze its influencing factors, so as to provide evidence for improving the hope level and quality of life of patients after THA. Methods? A cluster random sampling method was used to select 308 patients with bilateral THA for the first time who were admitted to two ClassⅢ Grade A general hospitals in Harbin from January 2017 to July 2018. Questionnaire surveys were conducted using Herth Hope Scale, the Second Edition of Beck Depression Scale and Social Support Scale. Results? The total score of hope level in elderly THA patients was (29.96±8.72). Among them, 76.62% (236/308) had moderate to high level of hope and the total score of depression was (19.20±11.75) which was at a moderate level. There were significant differences in the hope level of patients with different gender, place of residence, educational level, marital status and family per capita monthly income (P＜ 0.05). The results of multiple stepwise regression analysis showed that age, sex, family income per capita, depression, negative coping style, subjective support and objective support were the influencing factors of elderly THA patients' hope level (P ＜ 0.05). Conclusions? The THA patients' hope are at a medium level. Medical staff should pay attention to the relevant factors affecting the level of hope, adopt targeted nursing intervention measures, give psychological guidance to patients, provide social support, and further improve the level of hope of the THA patients.

Objective: To study the clinicopathologic features of dedifferentiated liposarcoma of extremities. Methods: Nine cases of dedifferentiated liposarcoma of extremities diagnosed at Beijing Jishuitan Hospital from 2009 to 2017 were selected. The histological features of cases of dedifferentiated liposarcoma of extremities were evaluated by HE and immunohistochemistry, together with the clinical and radiological features. Flourescence in situ hybridization(FISH) was used to detect MDM2 amplification. Results: They were located in the thigh (6 cases), calf (2 cases) and buttock (1 case). There were six females and three males. Patients′ age ranged from 61 to 79 years (mean 68 years). Histologically, there were two components, well-differentiated liposarcoma and dedifferentiated sarcoma with or without transitional lesions between them. The histology of dedifferentiated liposarcoma included undifferentiated sarcoma and fibrosarcoma. Heterologous elements such as bone and cartilage were present in two cases. Immunohistochemical study showed the tumor cells expressed vimentin, CDK4 and p16. MDM2 were positive in 6 cases (6/9) and p53 was positive in one case(1/9). CKpan was positive in the epithelioid differentiation area. S-100 protein was positive in the well-differentiated liposarcoma component. FISH showed the amplification of MDM2(6/9). Conclusions Dedifferentiated liposarcoma of extremities is very rare. The diagnosis should be combined with the histological characteristics and immunohistochemical results and differentiated from the other tumors and tumor-like lesions.

Objective: To study the clinicopathologic features of dedifferentiated chordoma. Methods: Four cases of dedifferentiated chordoma of sacrococcygeal region were collected at Beijing Jishuitan Hospital, from 2009 to 2014. HE and immunohistochemistry (EnVision method) were used to observe the clinical, radiological and histological features of dedifferentiated chordoma and to make the diagnosis and differential diagnosis. The literature was reviewed. Results: Four cases of dedifferentiated chordoma were all located in the sacrococcygeal region. The mean age at diagnosis was 57 years (range 49-64 years). There were 1 female and 3 males. Histologically, there were two components of conventional chordoma and dedifferentiated sarcoma with or without transitional area between them. The histology of dedifferentiated components includes undifferentiated sarcoma and fibrosarcoma. Immunohistochemical study showed that cytokeratin, epithelial membrane antigen, S-100, Brachyury were positive in the portion of chordoma while p53 were positive in the portion of undifferentiated sarcoma. INI1 and vimentin were both positive. Conclusions The dedifferentiated chordoma is very rare. The diagnosis should combine the histological characters and immunohistochemical results and should be differentiated from the sarcomatoid chordoma, poorly differentiated chordoma and other high grade sarcoma. p53 may play a role in the malignant transformation mechanism of chordoma.