BACKGROUND: Human pulp tissue has been known to be less, and exhibit poor tolerance to enzymatic digestion and less adherent cells after step-by-step digestion of trypsin and collagenase, thereby often leading to a failure of passage. Only several kinds of dental pulp cells with poor activity can be obtained by the tissue explant-collagenase digestion. OBJECTIVE: To investigate human dental pulp cells cultured in vitro by tissue explant method. DESIGN, TIME AND SETTING: A cytological observation was performed at Heping Campus and School of Stomatology, Jilin University from 2005 to 2007. MATERIALS: Healthy young human teeth extracted for orthodontic correction or impaction. METHODS: Pulp tissue from the third molar teeth was collected, cut into small blocks with a size of 1.0 mm×1.0 mm×0.5 mm under the infiltration of small amount of Dulbecco's modified eagle's medium, and then transferred into a 6-well plate containing culture medium for incubation in a 5% CO2 and saturated humidity atmosphere at 37 ℃. During the process of incubation, pulp tissue was adjusted at a density of 3-6 blocks/well, with an equal spacing of 0.5 cm and the 6-well plate was kept inverted. Three hours later, the 6-well plate was turned over to make tissue blocks adhering to the plate wall. Culture was continued after addition of 2 mL of culture medium. Culture medium was renewed every 4-6 days. After 6-15 days, cells emigrated from the edge of tissue blocks and call outgrowth appeared around each tissue block. When cells closed to confluency, a digestion procedure of 2.0-3.0 minutes (0.25% trypsin and 0.02% ethylenadiamine tetraacetic acid) was followed by passage culture at a proportion of 1: (2-3) in 25 mL of culture flasks. Purified fibroblast-like cells were gradually obtained from primarily cultured cells by repeated digestion and passage. MAIN OUTCOME MEASURES: Cellular morphology was identified by immunohistochemistry; secreted dental pulp cells were determined using alkaline phosphatase activity; the growth curves of human pulp tissue cells were depicted by MTT assay. RESULTS: Under an inverted phase contrast microscope, the obtained dental pulp cells were primarily typical fibroblasts with a long-shuttled appearance, well-rounded call body, uniform cytoplasm, round or oval nucleus, and clear nucleolus. Immunohistochemistry results showed call surface vimentin-positive, pan cytokeratin-negative, and alkaline phosphatase-posltive These cells were decreased after culturing 1 day, were slightly increased after 2 days, entered the logarithmic growth period and were markedly increased after 4 days, entered a platform period after 8 days, and began to decrease again after 9 days. The whole growth curve of cells appeared in "S" shape.CONCLUSION: The dental pulp cells isolated from human pulp tissue by tissue explant method can effectively proliferate end retain a poody differentiated state in vitro.

Objective To develop a clinical laboratory information system to execute information sharing.Methods The system realized information sharing between the third-party clinical laboratory facility and HIS with Oracle 10g database and Powerbuilder 9.0.Results The system implemented information sharing and informatized storing of the delivery specimen clinical laboratory results in the hospital.Conclusion The system fulfills seamless interface between the third-party clinical laboratory information system and HIS so as to provide the doctor and patient access to information and enhance the accuracy and timeliness of diagnosis.

Objective To elucidate the function way of micro RNA(miR)-155 in the differentiation of Th17 cells.Methods CD4+T cells were separated from mice spleens using MACS CD4+T cells separatinge kit and cultured with interleukins [interleukin (IL)-2, IL-23 and IL-6] which could induce CD4+ T cells differentiate into Th17 cells.IL-17 was detected by flow cytometry and enzyme linked immunosorbent assay (ELISA) after transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression levels of miR-155, IL-17A mRNA and Ets-1 mRNA were detected using fluorescent quantitation real-time quantitative polymerase chain reaction (RT-PCR).The si-Ets and miR-155 co-function for Th17 differentiation was analyzed.Data analysis was perfoemed using one-way analysis of variance (ANOVA) test and Dunnett test for pair-wise comparison and t test.P＜0.05 was considered to be statistically significant.Results The CD4+T cells were divided into four groups (the untreated control untreat group, the treatment control treat group, the miR-155 mimnics group and miR-155 inhibitor group).IL-17 was scarcely expressed and secreted in the untreated control untreat group.The cells expression of IL-17 were significantly different among the four groups (F=160.549, P＜0.01).The cells expressing of IL-17 were higher in the miR-155 mimics group (39.86±4.62)％ than those at the miR-155 inhibitor group (22.02±2.81)％, P＜0.01) and in the treated control treat group [(19.44±1.49)％, P＜0.01].The level of IL-17 was also significantly different among the four groups (F=260.813, P＜0.01).The level of IL-17 was higher in the miR-155 mimics group [(1 509±136) pg/ml] than that in the miR-155 inhibitor group [(923± 42) pg/ml, P＜0.01);and in the treated control group [(767±94) pg/ml, P＜0.01).The expression of miR-155 (12.53±0.80 vs 1.78±0.14, 7.16±0.62, 6.47±0.92, P＜0.01) and IL-17A mRNA (46.55±6.71 vs 1.01±0.19,15.62±1.26, 14.20±2.73, P＜0.01) was significantly higher than that in the other three groups, while the expression of Ets-1 mRNA was significantly lower (0.66±0.10 vs 1.19±0.04, 1.01±0.16, 1.37±0.27, P＜0.01).si-Ets-2 was screened because it markedly inhibited the expression of Ets-1 mRNA among the three designed siRNAs.The expression of IL-17A mRNA was higher (17.19±3.58 vs 10.08±0.76, t=-3.361, P=0.028) and the expression of Ets-1 mRNA was lower (0.27±0.01 vs 0.74±0.03, t=-30.275, P＜0.01) in si-Ets-2 group than that in si-Con group when si-Ets-2 or si-Con was co-transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression of Ets-1 protein was lower in si-Ets-2 group than that in si-Con group by Western blotting and the decrease was markedly obvious in the miR-155 mimics group.Conclusion miR-155 can induce CD4+T cells to differentiate into Th17 cells by inhibiting the gene expression of Ets-1.

BACKGROUND:The application of anterior cervical plate for anterior cervical discectomy and fusion wil induce some complications such as dysphagia after treatment.
OBJECTIVE:To observe the clinical efficacy of the new implant Zero-P system for cervical spondylosis patients and its effect on reducing complications after treatment.
METHODS:A total of 51 cervical spondylosis cases underwent anterior cervical discectomy and fusion using Zero-P system. Neurological function was evaluated by neck/arm visual analogue scale, neck disability index and Japanese Orthopaedic Association scores pretreatment, at 3 days, 3, 6 months, 1, 2 years post-treatment. The incidence of postoperative dysphagia was evaluated using dysphagia score. Anteroposterior&lateral and dynamics X-ray films were used to evaluate the efficacy of graft fusion and internal fixation-related complications.
RESULTS AND CONCLUSION:The patients were fol owed up for 6-30 months, averagely 15.4 months. After treatment, their incision achieved stage I healing. Pain was relieved and muscle force was restored in al patients after treatment. Visual analogue scale and neck disability index scores were apparently improved. Improvement rate of Japanese Orthopaedic Association score was 85.7%. At 2 days after treatment, seven patients experienced mild and moderate dysphagia. During fol ow up, implant subsidence was not observed. No complications, such as screw loose, breakage or fixator displacement, were found. Results suggested that the new cervical stand-alone anterior fusion device in anterior cervical discectomy and fusion procedure for the treatment of cervical spondylosis has offered an excellent clinical outcome. Its design has the advantages of cervical interbody fusion and anterior cervical plate. There were less relevant complications.

Objective : To examine the association between acute exposure to traffic-related air pollutants (TRAP) NOX and NO2 and outpatient visits of pediatric respiratory diseases. Methods : Data regarding outpatient visits to Department of Respiratory Diseases of Beijing Children's Hospital from 2015 to 2020 were collected, and the concentrations of nitrogen oxides (NOX), nitrogen dioxide (NO2) and other TRAP were collected from the surveillance sites assigned by the Peking University Health Science Center. A time-stratified case-crossover design was employed, and a conditional logistic regression model was created to examine the association between NOX and NO2 acute exposure and outpatient visits of pediatric respiratory diseases. Results : The daily mean outpatient visits of pediatric respiratory diseases were 571 (interquartile range, 554) person-times among children at ages of 0 to 14 years in Beijing Children's Hospital from 2015 to 2020, and the daily mean outpatient visits for upper respiratory tract infections (URI), bronchitis, and pneumonia were 265 (interquartile range, 282), 143 (interquartile range, 178) and 128 (interquartile range, 120) person-times, respectively. The daily mean concentrations of atmospheric NOX and NO2 were 67.8 (interquartile range, 50.7) and 49.3 (interquartile range, 30.7) μg/m3, respectively. Conditional logistic regression analysis showed the largest lagged effect of NOX and NO2 on pediatric respiratory diseases at cumulative lags of 0 to 7 days. An increase in NOX concentrations by an interquartile range resulted in the excess risks of URI, bronchitis and pneumonia by 6.87% (95%CI: 6.37%-7.38%), 7.25% (95%CI: 6.51%-7.99%), and 5.51% (95%CI: 4.69%-6.33%), and an increase in NO2 concentrations by an interquartile range resulted in excess risks of URI, bronchitis and pneumonia by 5.71% (95%CI: 5.12%-6.31%), 5.32% (95%CI: 4.51%-6.14%), and 4.83% (95%CI: 3.91%-5.75%), respectively. NOX and NO2 presented a more remarkable effect on outpatient visits of pediatric respiratory diseases among children at ages of over 5 years. Conclusion NOx and NO2 acute exposure may increase the outpatient visits of pediatric respiratory diseases.

Objective: To explore age-based clinical and immune parameters in Henoch-Schönlein purpura (HSP) to determine clinically useful markers reflecting disease characteristic. Methods: A cohort of 502 patients with HSP were enrolled into this retrospective study to evaluate their clinical and immune data. Results: Majority HSP cases occurred at age ≤14 years and showed significant immune imbalances of ESR, CD3⁺ cells, CD4⁺ cells, CD3^-CD16⁺CD56⁺ cells, CD4⁺/CD8⁺ cells, IgG, IgA, IgM, IgE, complements C3/C4 and ASO in the acute phase. Compared to patients aged >14 years, symptoms of joint were more frequent at disease onset in patients aged ≤14 years (20.8% vs 7.6%, χ²=13.547, P<0.001) , and involvement of digestive tract and joint were also more frequent (57.4% vs 33.8%, χ²=24.106, P<0.001; 55.9% vs 32.5%, χ²=23.768, P<0.001, respectively) , but not for involvement of kidney (21.4% vs 51.3%, χ²=42.440, P<0.001) . The patients aged ≤14 years had distinct immune state, reductions of CD3⁺ cells, CD4⁺ cells and IgG were more frequent than patients aged >14 years, also increase of ASO (33.1% vs 20.0%, χ²=6.656, P=0.010) , but not increase of IgA (2.6% vs 39.4%, χ²=15.582, P<0.001) . In addition, reduction of IgG and increase of IgE were positively associated with digestive tract involvement (P<0.001, P=0.001, respectively) , reduction of CD3⁺CD4⁺ cells and normal IgM were positively associated with joint involvement (P=0.004, P=0.003, respectively) , increase of CD3⁺CD8⁺ cells and normal CD3⁺ cells were positively associated with kidney involvement (P=0.032, P=0.014, respectively) . Conclusion HSP showed significant immune imbalance in the acute phase, patients between aged ≤14 and >14 years had distinct clinical and immune characteristic, and abnormal immune parameters were significantly associated with organ involvements.

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*

The constitutive androstane receptor (CAR, NR3I1) belongs to nuclear receptor superfamily. It was reported that CAR agonist TCPOBOP induces hepatomegaly but the underlying mechanism remains largely unknown. Yes-associated protein (YAP) is a potent regulator of organ size. The aim of this study is to explore the role of YAP in CAR activation-induced hepatomegaly and liver regeneration. TCPOBOP-induced CAR activation on hepatomegaly and liver regeneration was evaluated in wild-type (WT) mice, liver-specific YAP-deficient mice, and partial hepatectomy (PHx) mice. The results demonstrate that TCPOBOP can increase the liver-to-body weight ratio in wild-type mice and PHx mice. Hepatocytes enlargement around central vein (CV) area was observed, meanwhile hepatocytes proliferation was promoted as evidenced by the increased number of KI67