Objective To perfect the purification method of recombinant fusion protein of Hespintor (rHespintor) for increasing the protein extraction efficiency,and to investigate its effects on the proliferation,migration and invasion of hepatoblastoma cell line HepG2.Methods In the recombinant protein extraction,the inclusion body washing process was added and the protein purification buffer system was changed.BAPNA was used as the substrate.The inhibitory effect tof purified rHespintor on trypsin hydrolysis was detected.The blank group served as the control group.The MTT test,cell scratch wound healing test and tumor cell invasion test were performed to detect the effect of rHespintor on growth of hepatoblastoma HepG2 cells and its effect.Results The urea gradient washing on the inclusion body protein could effectively remove the vast majority of impure proteins from the targeted protein.After one-step purification,the target protein rHespintor exhibited a high inhibition effect of trypsin hydrolysis,and the inhibitory effect was exhibited a dose-dependent manner.After acting on hepatoblastoma HepG2 cells with rHespintor,the cell proliferation ability was inhibited,the migration ability was reduced and the number of invaded cells were significantly decreased.Conclusion rHespintor can significantly inhibit the proliferation,migration and invasion of hepatoblastoma cell line HepG2 cells in vitro.

Objective:To survey the working status of outpatient nurses in general practice departments of general hospitals.Methods:From March to April 2021, personal in-depth interviews were conducted with outpatient nurses from general practice residency training bases in Beijing. The thematic framework analysis method was used to analyze the interview data and refine the themes.Results:Fourteen nurses working in general practice outpatient clinics were interviewed in this study. Four themes were extracted from the interview data: (1) inadequate staffing of full-time outpatient nurses in general practice departments of comprehensive hospitals; (2) unclear job functions for outpatient nurses in general practice departments; (3) no standardized patient health education in the department; (4) no relevant training received systematically.Conclusion:General practice departments in general hospitals should setup full-time general practice nurse positions, clarify the job responsibilities, strengthen the relevant training and enhance core competency for nurses working in general practice department.

Adoptive cellular immunotherapy has been widely recognized in recent years due to its remarkable results, especially the success of CD19-specific chimeric antigen receptor (CAR) autologous T cell therapy for malignant hematoma. Previous studies have found the existence of tumor immune microenvironment, heterogeneous targets, and immunosuppressive receptors in solid tumors, which has led to the shortcomings of CAR-T treatment of solid tumors. This article proposes the methods to improve CAR-T cells to increase T cell infiltration, co-expression of cytokines and enzymes and modification of related receptors in order to enhance the anti-solid tumor activity of CAR-T, laying a theoretical foundation for the follow-up CAR-T cell treatment of solid tumors.

Objective:To investigate the past continuing medical education (CME) in pediatrics for general practitioners in community health centers in Longhua District, Shenzhen, so as to explore the willingness, needs and suggestions of general practitioners for CME in pediatrics.Methods:A mixed methods research were used in this study. Firstly, the purpose sampling method was used to select general practitioners in community health centers in Longhua District, Shenzhen for personal in-depth interviews. The interview contents were recorded and transcribed. The interview contents were coded by NVivo software. Theme frame analysis method was used for data sorting and theme extraction. Based on the results of qualitative interviews, the questionnaire was developed and distributed through the electronic questionnaire platform, and relevant questionnaire data were analyzed by Excel.Results:The results of qualitative research showed that after personal in-depth interviews with 10 general practitioners, the information reached saturation. Through repeated reading, induction and analysis of the interview data, four themes were extracted: ① previous pediatric training opportunities and satisfaction; ② willingness to participate in pediatric training; ③ the demand of training content, form and duration; ④ measures to encourage participation in training. In terms of quantitative research results, a total of 223 electronic questionnaires were collected, among which there were 219 valid questionnaires. Besides, 150 (68.5%) general practitioners said that they had no or only a few training opportunities in pediatrics CME in the past. There were some problems in the past CME training, such as unsystematic training (39 practitioners, 66.1%), narrow course coverage (30 practitioners, 50.8%), the content divorced from clinical practice (29 practitioners, 49.2%), monotonous teaching method (27 practitioners, 45.8%), etc. And 210 (95.9%) general practitioners were willing to participate in pediatric training, and 161 (73.5%) hoped that the form of training would be a combination of online and offline. In terms of pediatrics specialty content training needs, the general practitioners had higher demand for respiratory (188 practitioners, 85.8%), digestive (160 practitioners, 73.1%), infectious (145 practitioners, 66.2%) and dermatology (136 practitioners, 62.1%) specialty.Conclusion:There are still some problems in pediatrics CME, such as few training opportunities, unsystematic training, etc. In the future, we need to formulate systematic training plans and incentive measures according to the needs of general practitioners, to strengthen the pediatric professional training for general practitioners and improve their ability to receive children, in order to promote the implementation of hierarchical medical system for children.

Purpose To explore the predictive value of nomogram model for invasive breast cancer with axillary lymph node metastasis.Materials and Methods Retrospective analysis was made on 122 patients suspected to be breast cancer in the General Hospital of Ningxia Medical University from September 2020 to March 2022.According to whether there was axillary lymph node metastasis,all subjects were divided into 57 patients in the metastasis group and 65 patients in the non-metastasis group.All lesions were pathologically confirmed by surgery.The two groups received synthesis of magnetic resonance imaging(syMRI),dynamic contrast enhancement magnetic resonance imaging(DCE-MRI)and diffusion weighted imaging(DWI)scans.The syMRI parameters[including T1,T2,proton density(PD)],DCE-MRI time signal intensity curve,apparent diffusion coefficient(ADC)value of breast lesions were measured.Compared the difference of parameters between the two groups,and screened the independent risk factors of invasive breast cancer with axillary lymph node metastasis.Results Logistic regression results showed that Ki-67(OR=2.971,95%CI 1.306-6.762,P=0.009),lesion size(OR=1.652,95%CI 1.067-2.556,P=0.024),ADCratio(OR=1.685,95%CI 1.014-2.801,P=0.044),T2ratio(OR=3.015,95%CI 1.433-6.340,P=0.003),PDratio(OR=2.782,95%CI 1.471-5.262,P=0.002)were independent risk factors for invasive breast cancer with axillary lymph node metastasis.The comparison of the five models showed that the Logistic regression model had the best performance,with the area under curve of 0.729(95%CI 0.621-0.789),the accuracy,specificity and sensitivity were 70.65%,62.79%and 77.55%,respectively.The accuracy of the nomogram model was tested,and C-index=0.844,the accuracy of the nomogram model established was good,cut-off risk was 0.468,and the cut-off score was 143.50,which means that when the total score exceeds 143.50,the risk of axillary lymph node metastasis would be higher than 46.8%.Conclusion Nomogram model has a good predictive ability for invasive breast cancer patients with axillary lymph node metastasis.

Objective To investigate the differences of gut microbiota in patients with schizophrenia between onset and remission. Methods Twelve patients with schizophrenia in the stage of onset were selected as the episode group. Thirteen gender-, age-and BMI-matched patients with schizophrenia during the remission period were selected as the control group. The fecal specimens of the two groups were collected for high-throughput sequencing of the 16S rRNA gene, and the gut microbiota differences between the two groups were analyzed. Results The chao index and ace index of gut microbiota was lower in the episode group than in the control group (t=2.385,P=0.026; t=3.068,P=0.005). The relative abundance of Bacteroides was higher and the relative abundance of Prevotella was lower in the episodes group than in the control group (Z=-2.013, P =0.044; Z=-3.427, P=0.001). The relative abundance of the Prevotella was negatively correlated with the PANSS score (r=-0.577, P=0.003). Conclusion Schizophrenia in the stage of acute episode have altered gut microbiota compared with those in the stage of remission and the gut microbiota is significantly corrected with mental symptoms.

ObjectiveTo explore the inhibitory effect of water extract of Broussonetiae Fructus on hepatocellular carcinoma （HCC） induced by diethyl nitrosamine （DEN） in mice based on homologous phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B （PTEN/PI3K/Akt） signaling pathway. MethodThe primary HCC mouse model was constructed by intraperitoneal injection of DEN solution， and the HCC mice were randomly divided into model group， sorafenib group （0.01 g·kg^-1·d^-1）， low-dose Broussonetiae Fructus water extract group （0.9 g·kg^-1·d^-1）， medium-dose Broussonetiae Fructus water extract group （1.8 g·kg^-1·d^-1）， and high-dose Broussonetiae Fructus water extract group （3.6 g·kg^-1·d^-1）， with 10 mice in each group. Another 10 C57BL/6 mice were selected as a control group and intraperitoneally injected with an equal volume of normal saline. Mice were treated with different concentrations of Broussonetiae Fructus water extract when liver cancer-like white nodules appeared. sorafenib group was treated with sorafenib. The control group and model group were intraperitoneally injected with normal saline. The activities of alkaline phosphatase （ALP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， and γ-glutamyl transferase （γ-GT） in the serum of mice were detected by the biochemical analyzer. The expression levels of alpha-fetoprotein （AFP） and carcinoembryonic antigen （CEA） were detected by enzyme-linked immunosorbent assay （ELISA）. The degree of hepatocyte canceration and hepatocyte injury were observed by Hematoxylin-eosin （HE） and Masson staining. The proliferation of HCC cells was observed by immunohistochemical staining. The apoptosis of HCC cells in mice was observed by erminal-deoxynucleotidyl transferase mediated nick end labelling （TUNEL） staining. The expression levels of PTEN， PI3K， Akt， and p-Akt proteins related to the PTEN/PI3K/Akt signaling pathway were detected by Western blot. ResultCompared with the control group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the model group were significantly increased （P<0.01）. Carcinogenesis and inflammatory cell infiltration were obvious in liver tissue of mice， and a large number of blue collagen fiber hyperplasia was found. The number of Ki67 positive cells was significantly increased （P<0.01）， and the expression level of PTEN protein was significantly decreased， while PI3K and p-Akt protein expression was increased （P<0.01）. Compared with the model group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the medium-dose and high-dose Broussonetiae Fructus water extract groups were significantly decreased （P<0.05， P<0.01）. The degree of carcinogenesis and inflammatory cell infiltration in liver tissue were reduced， and the collagen fiber hyperplasia was significantly reduced. The number of Ki67 positive cells was significantly decreased， and the number of TUNEL positive apoptotic cells was significantly increased （P<0.05， P<0.01）. PTEN protein expression was increased， while p-Akt protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionThe water extract of Broussonetiae Fructus has a significant inhibitory effect on DEN-induced primary HCC in mice， and its mechanism may be related to the regulation of key protein expressions in the PTEN/PI3K/Akt signaling pathway.