Objective:To study the gene regulatory network during the osteogenic differentiation process of dental follicle stem cells (DFSCs)with bioinformatic analysis.Methods:The differentially expressed genes during the osteogenic diffrentiation process of DF-SCs were selected from the GEO Datasets.Then the relationship among the genes were analysed using DAVID and GeneMANIA data-base.Results:Numerous differentially expressed genes during the osteogenic differentiation process of DFSCs were obtained,289 genes with significance of different expression(P <0.05)were enriched into “cell differentiation”,“cell proliferation”,“skeletal sys-tem development”and “calcium signaling pathway”subgroups.ALPL,CTSK,TUFT1 ,TGFBR2,FHL2,ROR2,STC1 ,POSTN, ZBTB1 6,FRZB,PRELP and IGFBP5 were enriched into the “skeletal system development”subgroup.The 1 2 genes exhibited interac-tions,including co-expression,co-localization,genetic interaction and signal pathways.Conclusion:There is a complex molecular regulatory network among the differentially expressed genes during the osteogenic differentiation process of DFSCs.It is necessary to an-alyse the osteogenic differentiation process of DFSCs as a whole from WNT,TGFbeta,MAPK siganal pathyways,Homeobox transcrip-tion factors and osteogenic differentiation marker genes.

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Objective To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell(DC) transfected with interleukin(IL)-12 gene in vitro. Methods Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor ? were used to generate DC from cord blood in vitro. IL-12 fusion gene was cloned into a eukaryotic vector-pcDNA3.1(+). DC were transfected with IL-12-pcDNA3.1(+) using lipofectamine transfection method and electric transfection method respectively. Then polyethylene glycol mediated the fusion of transfected DC and ovarian carcinoma cells, and 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test was carried out to observe the immune response. Results DC were generated in vitro from cord blood and expressed high levels of costimulatory(50%) and MHC Ⅱ molecules(99%). RT-PCR showed that IL-12-pcDNA3.1(+) had been successfully transfected into DC. RT-PCR and immunofluorescence analysis showed that DC were fused with ovarian carcinoma cells successfully. Then the fusion cells of ovarian carcinoma cells to transfected DC, and the fusions of ovarian carcinoma cells to DC were cocultured with peripheral blood mononuclear cell respectively. MTT test showed that both fusions could induce cytolytic T cell activity and lysis of ovarian carcinoma cells,and the former effect was stronger. Conclusion The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.

Objective Detecting plasma level of circular RNA(circRNA)hsa_circ_0009024 in pulmonary tuberculosis(TB)patients,and evaluating its diagnostic value for TB.Methods From January 2016 to December 2016, a hosptial-based, case-control study was performed, which include 90 untreated active pulmonary tuberculosis patients(TB group),75 healthy people(healthy control)and 84 patient with other diseases(other disease group).Plasma level of circRNA hsa_circ_0009024 was detected by real-time quantitative polymerase chain reaction.Furthermore, the 90 patients with TB were divided into different subgroups according to cavity formation and the lung fields involvement: patients without lung cavity(55 cases)vs those with lung cavity(35 cases),patients with involvement of <2 lung fields(49 cases)vs≥2 lung fields(41 cases).Plasma levels of hsa _circ_0009024 of 41 TB patientswere monitored andcomparedbefore and after 3 months anti-TB therapy.The sensitivity and specificity of plasma hsa_circ_0009024 were analyzed by using the receiver operating characteristic(ROC)curve.The comparison between two groups was performed with Mann-Whitney U test and the comparison among multigroupswas conducted with Kruskal-Wallis H test.Results Plasma levels of hsa_circ_0009024 in TB patients[1.98(1.42, 2.71)]were significantly higher than healthy controls[1.03(0.78,1.33)]and other disease groups[1. 13(0.77,1.51)](H=76.58,P<0.0001).Plasma levels of hsa_circ_0009024 in cavity pulmonary TB patients were higher than pulmonary TB patients without cavity(U=392.50,P<0.0001).Plasma levels of hsa_circ_0009024 in TB patients with involvement of ≥2 lung fields were higher than <2 lung fields(U=590.50,P=0.0008).As compared to pre -treatment[2.01(1.41, 2.71)], the plasma hsa_circ_0009024 levels decreased significantly in 3 months[1.22(0.85,1.47)](U=292.50,P<0.0001)after anti-TB therapy.The area under the receiver operating characteristics curve(AUC)of plasma hsa_circ_0009024 in discriminating the patients with TB from normal controls, pneumonia patients and lung cancer patients were 0.841 and 0.811, respectively.Conclusion The hsa_circ_0009024 can be used as a potential biomarker in TB diagnosis and monitoring.

Objective: To investigate the pathologic features of gonadal tissues of disorders of sexual development (DSD) in children. Methods: Fifty-three cases of gonadal developmental disorders were collected from July 2015 to August 2017 at Guangzhou Women and Children′s Medical Center. Clinical manifestations, karyotypes, sex hormone levels, ultrasound imaging, histology and immunophenotype of gonadal tissues were analyzed. Results: The age of patients ranged from 7 months to 17 years with an average of (50.7 ± 47.1) months. Social genders of the patients included 32 males and 21 females. Forty-eight patients had abnormal sex hormone levels. Clinical presentations included: toward female genitalia in 25 cases, male genitalia tendency in 17 cases and ambiguous external genitalia in 11 cases. Hypospadias was seen in 31 cases and short stature was seen in 8 cases. Chromosomal karyotyping of peripheral blood revealed 23 cases of sex chromosome disorders, 22 cases of 46 XY disorders, of which 3 cases were 5α-reductase deficiency and 8 cases of 46 XX disorders. Ultrasound examination showed cryptorchidism in 30 cases, including 16 cases of unilateral, 14 cases of bilateral and 1 case presenting a huge pelvic tumor. A total of 97 gonadal tissues from 53 cases of DSD were examined, including 9 cases of unilateral and 44 cases of bilateral gonads. Microscopically, 55 gonads (56.7%) showed dysplastic testes including 17 unilateral and 19 bilateral gonads. Fourteen were streak gonads (14.4%) including 8 unilateral and 3 bilateral gonadal tissues. Nine streak gonad with epithelial cord-like structures (9.3%) were found, of which 5 were unilateral and 2 were bilateral lesions. Seven gonads were ovotestis (7.2%), unilateral in 5 cases (the other side of the gonads of ovary in 4 cases, 1 case of dysplastic testes) and bilateral in 1 case. Seven gonads showed follicular-rich ovarian tissue (7.2%). One case showed bilateral dysplastic testes with gonadoblastoma and ectopic adrenal cortex. One case of streak gonad showed epithelial cord-like structures and undifferentiated glandular tissue embedded in malignant mixed germ cell tumors (mixed gonadoblastoma, dysgerminoma, mature teratoma and yolk sac tumor). One case had testicular microlithiasis. Uterus and fallopian tube structures were found in 11 cases. Immunohistochemical stains were performed in 15 cases. D2-40, PLAP and CKIT were expressed in germ cells and Calretinin, WT1 and inhibin were positive in Setoli cells. SALL4 and OCT3/4 were positive in 3 cases. Inhibin highlighted interstitial Leydig cells in 2 cases. GPC3 was positive in yolk sac tumor component. Conclusions Gonadal dysgenesis presents a broad spectrum of gonadal phenotypes with variable degrees of differentiation. The development of bilateral gonadal tissues has certain variability. Chromosomal karyotypes have no correlation with gonadal phenotypes. Accurate histopathologic diagnosis of gonadal dysgenesis plays an important role in the treatment and prognosis of the patient.

Objective:To compare the value of NoSAS score, STOP-BANG questionnaire (SBQ) and Epworth Sleepiness Scale (ESS) in assessing the risk of obstructive sleep apnea hypopnea syndrome (OSAHS) in patients with respiratory disease (RD).Methods:The clinical data, NoSAS, SBQ and ESS scores of 190 patients who underwent overnight polysomnography (PSG) were collected. According to the receiver operating characteristic (ROC) curve, with different apnea-hypopnea index (AHI) as the judgment cutoff, the sensitivity, specificity, positive predictive value, negative predictive value, diagnostic odds ratio (DOR) and accuracy of the three scales were compared.Results:With AHI ≥5 times/h as the cutoff, the area under the ROC curve (AUC) of NoSAS and SBQ were 0.833 and 0.729, respectively, indicating that both have predictive value for mild OSAHS. Among them, NoSAS had a larger DOR value (16.150), indicating that NoSAS had the higher accuracy in assessing the risk of mild OSAHS. When AHI>15 times/h was used as the cutoff, the AUC value of NoSAS was 0.704, indicating that it has predictive value for moderate OSAHS. When AHI>30 times/h was used as the cutoff, the AUC value (0.706) and DOR value (6.527) of SBQ were high, indicating that it has predictive value and good accuracy for severe OSAHS. The SBQ is more sensitive than NoSAS and ESS when evaluating patients at high risk for OSAHS ( SBQ≥3). Conclusions:When evaluating the risk of mild and moderate OSAHS in RD patients, NoSAS is better than SBQ and ESS, and when evaluating severe OSAHS, SBQ is better than NoSAS and ESS. In clinical work, appropriate predictive tools should be selected according to the actual situation to assess the risk of OSAHS, so as to formulate and implement early intervention plans based on the assessment results.

OBJECTIVETo summarize the clinicopathologic features of neuroblastic tumors (NT), and to explore the prognostic significance of MYCN amplification in NT.

METHODSThe clinicopathologic data of 267 NT were reviewed. MYCN gene amplification was detected by fluorescence in situ hybridization (FISH) in 119 cases and the relationship with pathological characteristics and prognostic significance were analyzed.

RESULTSThe study included 267 cases of children NT from patients aged from 1 day to 13 years (median 27 months). The male to female ratio was 1.43. There were 38 cases (14.2%), 43 cases (16.1%), 71 cases (26.6%), and 115 cases (43.1%) of INSS stages I, II, III and IV respectively.Favorable histology group had 157 cases (59.9%); unfavorable histology group had 110 cases (40.1%).Of the 119 NT cases with MYCN FISH performed, 18 cases (15.1%) showed amplification and the signal ratio of MYCN to CEP2 was 4.08-43.29. One hundred and one cases of non-amplified MYCN included MYCN gain in 79 cases (66.3%) and MYCN negative in 22 cases (18.5%). MYCN expression showed significant difference (P = 0.000) between ages, gender, NT type and MKI, but not INPC and clinical stage (P > 0.05).Of the 18 cases with MYCN amplification, 3 were undifferentiated, and 15 poorly differentiated; 17 had high MKI and one moderate MKI. All 18 cases were in unfavorable histology group; the overall survival rate was 3/18, with an average survival time of (17.9 ± 2.4) months.Of the 101 MYCN non-amplification cases, the overall survival rate was 68.3% (69/101), with an average survival time of (29.8 ± 1.3) months. Survival analysis showed the cases with MYCN amplification had worse prognosis (P < 0.05).

CONCLUSIONSNT were commonly diagnosed in early ages and easily to metastasize. Most of cases with favorable histology. The cases of MYCN amplification showed unfavorable histology, and the majority cases with high MKI; The patients with MYCN gene amplification had poor prognosis.

Adolescent ; Cell Differentiation ; Child ; Child, Preschool ; Female ; Gene Amplification ; Humans ; In Situ Hybridization, Fluorescence ; Infant ; Male ; N-Myc Proto-Oncogene Protein ; Neuroblastoma ; genetics ; mortality ; pathology ; Nuclear Proteins ; genetics ; Oncogene Proteins ; genetics ; Prognosis ; Survival Analysis ; Survival Rate

Objective To investigate the health status of centenarian hospitalized patients and analyze the risk factors for in-hospital death in Nanjing district.Methods All centenarians hospitalized patients who were discharged from wards of 10 upper first-class general hospitals in Nanjing district during the past five years were retrieved from their hospital information systems.Then,a retrospective study was performed on centenarians' data of general information,laboratory test results,Charlson comorbidity index (CCI),neutrophil to lymphocyte ratio (NLR) and shock index(SI),etc.were calculated and collected.Relevant risk factors for in-hospital death were analyzed by multivariate logistic regression analysis.Results A total of 156 patients aged 100 years and over,with an average age of (101.0±2.1)years,were enrolled during the past 5 years.The top 3 admitting diagnosis for the patients were pulmonary infection(30.1％,47/156 cases),coronary heart disease(10.9％,17/156 cases)and cerebrovascular disease(7.1％,11/156 cases).Fifty patients died during hospitalization,with a mortality of 32.1％ (50/156).Pneumonia was the most common admitting diagnosis(40.0％,20/50 case).Among causes of death,the combined admitting diagnosis with dementia,chronic renal insufficiency,one or more basic disease were significantly associated with death.There were statistically significant differences between bad vs.good vs.indifferent prognosis in heart rate,shock index,leukocyte count,neutrophil count,NLR,hemoglobin,albumin,albumin/globulin,fasting blood glucose,blood urea nitrogen,serum creatinine,C-reactive protein(CRP)and CCI levels.Multivariate logistic regression analysis suggested that NLR≥13.18,fasting blood glucose ≥7.56 mmol/L,blood urea nitrogen ≥20.74 mmol/L,CRP≥65 mg/L and CCI≥3 might be predictors for in-hospital death in the cohort(OR =48.91、3.43、1.22、6.55、1.55,all P＜0.05).Conclusions Pulmonary infection is the most common reason for admission and the cause of death in centenarian inpatients.Comorbidities increase the risk of death.To lower in-hospital mortality,CCI and other assessment indicators should be used to strengthen the comprehensive assessment and chronic disease management of hospitalized centenarians.Infectious diseases should be prevented beforehand.

OBJECTIVE：To optimize the p reparation technology of citronellol submicroemulsion. METHODS ：The content of citronellol in Citronellol submicroemulsion was determined by HPLC. Citronellol submicroemulsion by high-speed shearing dispersion-high pressure homogenization method ，with centrifugation stability constant （ke） and particle size were used as evaluation indexes. Its formulation and preparation technology were optimized and validated. Drug-loading amount and encapsulation rate of the preparation were detected. RESULTS ：The linear range of citronellol were 4-64 μg/mL（R 2＝0.999 9）. RSDs of precision ，stability（24 h）and reproducibility tests were all lower than 3％. The recoveries were 97.64％-101.97％（RSD＝ 2.28％，n＝3），97.71％-99.50％（RSD＝1.29％，n＝3），96.87％-101.48％（RSD＝2.86％，n＝3）. The optimal formulation included that total weight of soybean oil and medium chain triglycerides （1 ∶ 1，g/g）was 3.75 g，1.2％ soybean phospholipid was 0.6 g， cholesterol was 0.06 g，citronellol was 1.25 g，0.6 ％ sodium oleate was 0.3 g，15-hydroxystearic acid polyethylene glycol ester was 0.75 g，poloxamer 188 was 0.75 g，water added to 50 mL. After prepared by optimal technology at 4 ℃ which contained shearing speed of 13 000 r/min，lasting for 5 min， primary emulsion was adjusted to pH 7 with dilute hydro- chloric acid ，and homogenized with 600 Bar high pressure for 1434412440@qq.com 5 min. The parameters of Citronellol submicroemulsion accor- ding to optimal formulation and technology contained mean particle size of （91.05±0.26）nm，PDI of （0.20±0.01）， Zeta-potential of （－30.86±0.39）mV，average content of 649511230@qq.com citronellol（100.21±0.01）％，the drug-loading amount was （2.481 7 ± 0.000 7） mg/mL，the encapsulation rate was （99.27 ± 0.03）％ . CONCLUSIONS ：The optimal formulation and technology is stable and feasible.