Objective : To investigate the expression and clinical significance of kynurenine-3-monooxygenase (KMO) in peripheral blood mononuclear cells ( PBMC ) ，synovial tissue ，and fibroblastic-like synovial cells (FLS) of rheumatoid arthritis (RA) patients． Methods : Peripheral blood samples from 25 healthy control ( HC) individuals and 25 patients diagnosed with RA were collected ，and real-time fluorescence quantitative PCR and Western blot were used to detect KMO gene and protein expression in PBMC of RA and HC groups，and to analyze the correlation between the expression level of the KMO gene in the PBMC of the RA patients and the indexes of the laboratory tests．Meanwhile，immunohistochemistry and immunofluorescence were used to detect KMO expression in synovial tissue and FLS in RA and HC groups． Results : ① KMO gene and protein expression in PBMC of RA group were higher than that of HC group，and the difference was statistically significant (P＜0. 001) ．② The level of KMO gene expression in PBMC of RA group was positively correlated with disease activity index 28 score，blood sedimentation，and rheumatoid factor (rs = 0. 417，P = 0. 038 ; r = 0. 545，P = 0. 005 ; rs = 0. 433，P = 0. 031) ， and had no correlation with C-reactive protein and anti-cyclic citrullinated polypeptide antibody．③ KMO expres- sion in synovial tissue of RA group was higher than that of HC group，and the difference was statistically significant (P＜0. 01) ; KMO expression in FLS of synovial tissue of RA group was higher than that of HC group，and the difference was statistically significant (P ＜0. 001) ． Conclusion KMO expression increases in PBMC ，synovial tissue and FLS of RA patients，and the level of KMO gene expression is correlated with the disease activity of RA patients，suggesting that KMO may promote the course of RA．

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective To provide an animal model for studying the effect of TDO2 on the function of macrophages on the occurrence and development of diseases by constructing macrophage-specific tryptophan,2,3-dioxygenase(TDO2)gene knockout mice.Methods TDO2flox/flox Lyz2-iCre+mice were constructed based on Cre/LoxP sys-tem.The genotypes of mice were identified by PCR amplification and agarose gel electrophoresis.Western blot and immunofluorescence were used to verify the effect of TDO2 knockdown in mouse macrophages.The spontaneous le-sions in major tissues and organ were observed by HE stainings.Results The results of genotype identification showed that the mice with only one band at 407 bp or 408 bp for the flox amplification product and one band at 543 bp for the Cre amplification product were TDO2flox/flox Lyz2-iCre+mice.Western blot results showed that TDO2 ex-pression in bone marrow-derived macrophages(BMDMs)of TDO2flox/flox Lyz2-iCre+mice decreased compared with TD02flox/flox mice(P＜0.01).Immunofluorescence results showed that TDO2 expression in peritoneal macrophages and BMDMs of TDO2flox/flox Lyz2-iCre+mice decreased compared with TDO2flox/flox mice.HE staining showed no sig-nificant differences in cell morphology in the liver,brain,kidney and spleen tissues of TDO2flox/flox Lyz2-iCre+mice compared to TDO2flox/f1ox mice.Conclusion TDO2flox/flox Lyz2-iCre+mice is successfully constructed,providing a more precise experimental animal model for subsequent in-depth study of the role and mechanism of TDO2-regulated macrophage activation in disease.

Nearly a quarter of the world's population is infected with Mycobacterium tuberculosis and remains long-term asymptomatic infection. Rv2626c is a latent infection-related protein regulated by DosR of M. tuberculosis. In this study, the Rv2626c protein was prokaryotically expressed and purified, and its immunobiological characteristics were analyzed using RAW264.7 cells and mice as infection models. SDS-PAGE and Western blotting analysis showed that the Rv2626c-His fusion protein was mainly expressed in soluble form and specifically reacted with the rabbit anti-H37RV polyclonal serum. In addition, we found that the Rv2626c protein bound to the surface of RAW264.7 macrophages and up-regulated the production of NO. Moreover, the Rv2626c protein significantly induced the production of pro-inflammatory cytokines IFN-γ, TNF-α, IL-6 and MCP-1, and induced strong Th1-tendency immune response. These results may help to reveal the pathogenic mechanism of M. tuberculosis and facilitate the development of new tuberculosis vaccine.
Animals ; Mice ; Rabbits ; Mycobacterium tuberculosis/genetics* ; Tuberculosis ; Antigens, Bacterial ; Cytokines ; Immunity, Cellular

Objective:To explore the application of laparoscopic training in the standardized residency training of pediatric general surgery.Methods:A total of 36 rotating residents who received the standardized residency training in the department of pediatric general surgery of Shenzhen Children's Hospital from January 2017 to December 2019 were selected for laparoscopic training. The training content includes the study and training of laparoscopic theory knowledge, laparoscopic surgery video, skills operation of simulated operating system. Assessment was conducted before and after the training, and statistical analysis was performed to compare the difference of scores before and after the training. The self-evaluation of resident's learning efficiency and the satisfaction with teachers were investigated by questionnaires. SPSS 20.0 was used for paired t-test. Results:After receiving the standardized residency training of pediatric general surgery, the theoretical knowledge and cognition of the application on pediatric general laparoscopic surgery were significantly improved among the 36 residents. The time that the skills spent in vitro simulation box during the simulated operation training was significantly reduced after training, with a statistically significant difference ( P < 0.05). According to the questionnaire survey, resident's satisfaction with self-evaluation of learning efficiency was 97.22% (35/36), their satisfaction with teachers was 94.44% (34/36), and their satisfaction with teaching curriculum was 100.00% (36/36). Conclusion:Laparoscopic training can effectively improve the clinical practice ability of pediatric general surgery residents, which is worthy of promotion.

Objective:To understand the current situation of the demand for Internet plus nursing service among nurses at different levels, and analyze the influencing factors, so as to provide reference for the improvement of relevant national policies.Methods:A total of 19 762 in-service nurses from 32 provinces in China were investigated by cross sectional survey in April to May 2019. The demand for Internet plus nursing services was investigated, including 5 dimensions, including institutional demand, safety demand, demand for fees, quality demand and training needs.Results:Among the 19 762 nurses surveyed, there were 135, 4 736, 8 847, 4 827 and 1 217 nurses at N0-N4 levels respectively. The safety needs of nurses ranked first(4.68±0.52). The institutional needs of N3 and N4 nurses were the most significant, and the safety needs of N0-N2 nurses were the most significant; Training demand was the third demand of nursing staff at all levels. Multiple linear regression analysis showed that education and professional title were the factors that affected nurses′ demand for Internet plus nursing service( P<0.01). Conclusions:Nurses at different levels have a high and diversified demand for Internet+ nursing services.

Objective:To investigate the demand of patients for home care services under the background of "Internet+" and analyze its influencing factors of their participation status.Methods:A total of 494 patients from 7 ClassⅢ hospitals in Beijing, Guangzhou and Changsha from April to June 2019 were investigated by questionnaires. The research tools included general data questionnaire and questionnaire of patients' demands or "Internet+" home care service.Results:A total of 461 valid questionnaires were collected. The degree of patients' demand for "Internet+" home care services in descending score order was diagnosis and treatment information security [ (4.64±1.69) points], service capacity [ (4.62±1.77) points], health guidance [ (4.34±3.06) points], specialty nursing [ (4.24±3.79) points]. The results of binary Logistic regression analysis showed that monthly income, medical insurance type, main caregivers, and the number of diseases suffered by family members were the influencing factors of patients' participation in "Internet+" home care services ( P<0.05) . Conclusions:Patients have a realistic demand for "Internet+" home care services. It is recommended to establish a nursing service system based on patient needs, strengthen patients' information security, expand the radius of nurses' door-to-door service, enrich the list of service items and provide patients with professional and scientific services.

Objective: To construct a three-dimensional (3D) finite element model and analyze the biomechanical characteristics during mandibular second molar mesialization using customized lingual appliances and mini-implant. Methods: One adult student volunteer from The First Affiliated Hospital of Zhengzhou University with lower left first molar extraction was selected. After CT scanning, Mimics, Geomagic, ANSYS were employed to develop a 3D finite element model including customized lingual brackets, stainless steel lingual arch wire, buccal buckles, lower dentition, periodontal ligaments and alveolar bone. Four different loading methods (1. the force of 1.470 N loaded at the lingual side only; 2, 3, 4. the forces of 0.490, 0.735 and 0.980 N loaded at both buccal and lingual sides, respectively.) were included. The initial displacements of the lower second molar and stress distribution in the periodontal ligaments were analyzed. Results: More uniform stress distributions in the periodontal membrane of mandibular left second molar were found when the mesial force were loaded at both buccal and lingual sides than the force loaded at lingual side only. More inclination of the second molar was observed when force loaded at both sides than at lingual side only. With the values of the force increased, the mesial inclination potential of the second molar, the initial movement of the second molar and the stress distribution in the periodontal membrane increased. Under the force of 0.735 N at both buccal and lingual sides, mesially inclined initial displacements of the mandibuar second molar were observed and the stress values of von Mises was in the suitable area. Conclusions Less second molar rotation was found when the force loaded at both buccal and lingual sides than loaded at lingual side only. However, force loaded at both sides would increase the measial inclination potential of the second molar.

OBJECTIVE:To provide reference for the supply of emergency drugs. METHODS:The data of emergency drugs in the inpatients of our hospital were collected from hospital information system during 2012-2016,including the proportion and age of emergency patients,drug,clinical departments and disease types. The trend of emergency drugs was analyzed by using SPSS 16.0 software. RESULTS:The proportion of emergency patients and discharged patients were increasing year by year. The number of patients in <10 year-old and 10-19 year-old showed a declining trend year by year. The number of patients in 30-39 year-old,50-59 year-old and >60 year-old showed an increasing trend year by year. Among emergency drugs,the use frequency of Heparin sodium injection,Dexamethasone sodium phosphate injection and Flumethasone injection showed a declining trend year by year;the use fre-quency of Hemagglutination for injection,Penehyclidine ether injection,Atracurium besylate injection and Propofol injection showed an increasing trend year by year. Among clinical department,the number of pediatric patients showed a declining trend year by year;the number of patients in obstetrics department,emergency department,cardiology department,micro hand surgery department and pediatric surgery department showed an increasing trend year by year. Among emergency disease types,the number of patients with re-spiratory system disease,infectious and parasitic diseases showed a declining trend year by year;the number of patients with circula-tion system disease,injury and poisoning,certain other consequences of external factors,digestive system diseases,pregnancy,child-birth and puerperium disease as well as some cases originated in the perinatal period showed an increasing trend year by year. CON-CLUSIONS:With the change of the proportion,age and disease type of emergency patients,the type and utilization rate of emergen-cy drugs have changed. It is necessary to adopt effective measures to guarantee the supply of emergency drugs.