Objective: To analyze the physical and chemical properties of human gastric factor by bioinformatics method , and to express and purify the protein. Methods: Protein online analysis software was used to predict the physical and chemical properties of gastric intrinsic factor and analyze their hydrophilicity and hydrophobicity. The online tool was used to predict and analyze the human gastric factor signal peptide and its subcellular location . The eukaryotic expression recombinant plasmid pcDNA3 . 1 ( + ) _human GIF_His tag was constructed , and the gastric intrinsic factor was purified by nickel column after expression in HEK293F cells . The purity and activity of purified gastric intrinsic factor were verified by SDS_PAGE , Western blot and indirect ELISA . Results: Gastric factor contained 417 amino acids and was a hydrophilic acid stable protein . It was a secreted protein with Sec original signal peptide . pcDNA3 . 1 ( + ) _human GIF_His tag recombinant plasmid was successfully constructed and soluble expression was obtained in HEK293F cell expression system. Conclusion The eukaryotic source of human gastric intrinsic factor is successfully prepared , and the bioinformatics results show that the protein is a hydrophilic acid stable secreted protein , laying a foundation for the subsequent use of this protein as an immunogen and protein calibrator to construct an immunoassay for gastric factor.

As an assistive device to restore lower limb athletic ability,human lower limb rehabilitation robots are becoming increasingly important in the field of rehabilitation and clinical applications.With the advancement of science and technology,both domestic and foreign research in this field has been developed.This study provides a detailed overview of the development of lower limb rehabilitation robots and reviews the current status of clinical applications,with a focus on mechanism research,from the perspectives of degrees of freedom,workspace,singularity,gait simulation,kinematic simulation and human-robot interaction,etc.The results show that domestic research on lower limb rehabilitation robots focus on the design and optimization of the mechanism configuration,while foreign research focus on the improvement and innovation of the control system and training mode based on human-computer interaction.Finally,the current state of research is combined with an outlook on future trends.

Objective:To explore the application value of histogram and texture feature analysis based on CT images in distinguishing long bone osteosarcoma (OS) from Ewing sarcoma (ES).Methods:A retrospective collection of 25 patients with long bone osteosarcoma and 25 patients with Ewing sarcoma confirmed by surgery and pathology in National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Qilu Hospital of Shandong University and Nanjing Drum Tower Hospital, Nanjing University Medical School, from March 2018 to May 2023 was conducted. All patients were randomly divided into a training set (21 cases of OS and 19 cases of ES) and a validation set (4 cases of OS and 6 cases of ES) in an 8∶2 ratio. The region of interest (ROI) on CT images to extract texture feature parameters was manually sketched. Random forest and least absolute shrinkage and selection operator (LASSO) algorithm were used for feature screening. Logistic regression (LR), random forest (RF), support vector machine (SVM) and K-nearest neighbor (KNN) classifiers were used to establish models respectively. Receiver operating characteristic (ROC)curve was drawn and area under the curve (AUC) was calculated to evaluate the diagnostic efficiency of the four models.Results:A total of 100 texture parameters were extracted from CT images, and 8 feature parameters (maximum 3D diameter, 10th percentile, kurtosis, maximum pixel intensity value, inverse normalization, grayscale level variance, long range high grayscale emphasis, and low grayscale area emphasis) were obtained through screening. Four classifiers were used to establish models, and the AUC values of the four models (LR, RF, SVM, KNN) in the validation group were 0.92, 0.79, 0.83, and 0.73, respectively. LR and SVM classifier algorithm trains models had high diagnostic efficiency, with an accuracy of 90%, sensitivity of 83%, specificity of 100%, and AUC of 92% for the LR classifier validation set; the accuracy of SVM classifier validation set was 80%, sensitivity was 67%, specificity was 100%, and AUC was 83%.Conclusions:LR and SVM models have high value in distinguishing OS and ES.

Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the metal organic framework (MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS). Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked phosphate buffer (PB) model, faster modification trends were monitored in the spiked serum and patients' sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients' sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.

Dynamic tracking analysis of monoclonal antibodies(mAbs)biotransformation in vivo is crucial,as certain modifications could inactivate the protein and reduce drug efficacy.However,a particular chal-lenge(i.e.immune recognition deficiencies)in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen.To address this limitation,a multi-epitope affinity technology utilizing the metal organic framework(MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region(CDR)mimotope peptide(HH24)and non-CDR mimotope aptamer(CH1S-6T)onto the surface of MOF@Au nanocomposite.Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited signifi-cantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope af-finity technology.Moreover,the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry(LC-QTOF-MS).Therefore,multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb.Particularly,the novel bioanalytical platform can be suc-cessfully used for the tracking analysis of trastuzumab modifications in different biological fluids.Compared to the spiked phosphate buffer(PB)model,faster modification trends were monitored in the spiked serum and patients'sera due to the catalytic effect of plasma proteins and relevant proteases.Differences in peptide modification levels of trastuzumab in patients'sera were also monitored.In summary,the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb,contributing to improved understanding and paving the way for future research and clinical applications.

Objective:To investigate the effects of TRIB3 activation of Wnt/β-catenin signaling pathway on the growth and pro-liferation of laryngeal carcinoma TU686 cells in vitro and expression of peripheral immunosuppressive molecules in transplanted mice.Methods:Protein and RNA expressions of TRIB3 were detected in vitro cells(human immortalized epidermal cell line HaCat and laryngeal carcinoma cell line TU686)and tissues(laryngeal carcinoma and adjacent tissues),respectively.Laryngeal carcinoma TU686 cells were cultured in vitro and divided into negative control group(NC group)and TRIB3 knockdown group(sh-TRIB3 group),total protein and RNA of cells were extracted to verify the expression level of TRIB3 in two groups.After successful verifica-tion,proliferation ability of TU686 cells was detected by CCK-8,colony cloning and flow cytometry.Protein expression levels of Wnt,Cyclin-D1,C-myc,β-catenin and p-β-catenin in two groups were detected by Western blot.Correlation analysis verified the correla-tion between TRIB3 and Wnt,Cyclin-D1,C-myc,β-catenin,p-β-catenin protein expressions.TRIB3-low expressing nude mouse transplanted tumor model(TRIB3 sgRNA group)was constructed by knockdown the TRIB3 core plasmid,and a parallel control group(Control sgRNA group)was set up,tumor growth volume and weight were observed,and serum immunosuppressive molecules expres-sions was determined by ELISA.Results:Compared with HaCat cells and normal paracarcinoma tissues,TRIB3 was highly expressed in TU686 cells and laryngeal carcinoma tissues.Compared with negative control group,proliferation ability of TU686 cells was signifi-cantly inhibited after TRIB3 knockdown,and cell growth was blocked in G1/S phase.Expressions of Wnt,Cyclin-D1,C-myc andβ-catenin protein in Wnt/β-catenin signaling pathway were decreased significantly,while expression of p-β-catenin was increased significantly.TRIB3 was significantly correlated with protein expression levels of Wnt,Cyclin-D1,β-catenin and p-β-catenin.The in vivo results showed that compared with Control sgRNA group,tumor growth volume and weight of mice in TRIB3 sgRNA group were significantly decreased,and expressions of serum immunosuppressive molecules IL-4,IL-6,IL-10,TGF-β and PGE2 were signifi-cantly decreased.Conclusion:TRIB3 is highly expressed in TU686 cells,and TRIB3 can inhibit growth and proliferation of TU686 cells and transplanted tumors by activating Wnt/β-catenin-related signaling pathways,and reverse tumor immunosuppressive microen-vironment,suggesting that TRIB3 may be an effective target for laryngeal cancer.

Objective:To investigate the current management of nosocomial infection at medical institutions of all levels in Changzhou, so as to provide basis for standardizing nosocomial infections control of hospitals within a medical alliance.Methods:An electronic questionnaire was customized for online survey of 91 hospitals affiliated to eight regional medical alliances in Changzhou city in March 2019. The survey covered such aspects as general conditions of the hospital, profile of nosocomial infection control administrators and other staffing, supervision of hospital nosocomial infection programs, and training needs, as well as outstanding problems and suggestions.Frequency number and percentage represent enumeration data, and χ2 test was used to analyze the in-group differences of medical institutions of three levels. Results:Tertiary public hospitals were superior to the secondary and primary hospitals in organizational structure, professional staffing and target monitoring, with the differences of statistical significance( P<0.05). The most urgent training needs of medical institutions at all levels were knowledge in determination and reporting of infectious diseases/nosocomial infection/infection outbreaks; top imperatives and recommendations were development of operation rules for primary medical institutions and standardization of workflows. Conclusions:Staff of primary medical institutions need capacity building in nosocomial infection control; primary hospitals are equipped with incomplete nosocomial infection control information platform; key departments in general lack homogenous management. Tertiary hospitals are encouraged to play leadership in medical alliances in achieving standardized, homogenous and informationized nosocomial infection control within the medical alliances.

Burkitt lymphoma (BL) is an aggressive B-cell non-Hodgkin lymphoma which often occurs in children. The cure rate of BL is significantly increased with the wide application of short-duration and high-intensity immunochemotherapy, however, chemotherapy-related adverse reactions are still a big problem in adult patients and human immunodeficiency virus (HIV) positive patients. Meanwhile, the absence of effective immune targeting new drugs in patients with relapsed and refractory BL needs to be solved clinically. How to optimize the therapeutic regimen to reduce the chemotherapy-related adverse reactions and develop effective immune targeting new drugs is the hot spot in current research. This paper reviews the treatment progress of BL according to the 59th American Society of Hematology (ASH) Annual Meeting.

Objective:To explore the expression and biological functions of miR-346 in nasopharyngeal carcinoma.Methods:63 cases nasopharyngeal carcinoma tissue and 34 cases nasopharyngeal non-cancer tissues were collected,the miR-346 expression were detected by Real-time PCR between nasopharyngeal carcinoma tissue and nasopharyngeal non-cancer tissues,6 strains of human nasopharyngeal carcinoma cell lines and 1 strain of normal nasopharyngeal epithelial cell immortalized NP69.Two cell lines with middle expression levels in human nasopharyngeal carcinoma cells lines were selected,and transfected into miR-346 inhibitor,the control group (NC group) were with negative control plasmid transfection,miR-346 expression in two groups were detected by Real-time PCR,the proliferation,apoptosis were detected by Brdu-ELISA and flow cytometry,the migration and invasion were detected by Transwell Chambers experiments.Results:Compared with the nasopharyngeal non-cancer tissues,the miR-346 expression in nasopharyngeal carcinoma tissues was significantly increased (P =0.000);compared with the normal nasopharyngeal epithelial cells NP69.the miR-346 expression in human nasopharyngeal carcinoma cell lines was significantly increased (P＜0.05).The CNE-1 and CNE-2 were chose,after the miR-346 inhibitor transfection,the miR-346 expression were significantly lower compared with NC group,the difference was statistically significant (P＜0.05).The proliferation of two kinds of nasopharyngeal carcinoma cell CNE-1 and CNE-2 were restrained after transfection,the difference showed statistically significant 3 days after transfection (P ＜ 0.05).The apoptosis increased significantly,and the migration cell numbers and invasion cell numbers decreased significantly compared with NC group,the differences were statistically significant (P＜0.05).Conclusion:miR-346 is overexpression in nasopharyngeal carcinoma,down-regu-lation of miR-346 inhibits the proliferation,migration,invasion,promotes the apoptosis,miR-346 may act as a oncogene and play an important role in the pathogenesis of nasopharyngeal carcinoma.

Objective To investigate the effects of miR-100 on the proliferation of MIA PaCa-2 and CFPAC-1 cells through targeting fibroblast growth factor receptor 3 (FGFR3).Methods miR-100 expression levels in 17 cancer tissues and 17 nonmalignant tissues were examined by Real-time PCR.The effect of miR-100 overexpression on cell proliferation was examined by CCK-8 assay in vitro.Luciferase assay was used to confirm that miR-100 could directly target FGFR3.Real-time PCR and Western blot were used to examine the expression of FGFR3 in miR-100 overexpressing pancreatic cancer cells.The predicted target gene of miR-100,FGFR3,was downregulated by siRNA,and its effect on cell proliferation was also examined.Cell proliferation was analyzed using CCK-8 and Edu assay.Results miR-100 was lowly expressed in pancreatic cancer tissues (P ＜ 0.05).In pancreatic cancer cells,the transfection of lv-miR-100 was able to upregulate the endogenous expression of miR-100 and inhibit the cell proliferation (P ＜0.05).Luciferase assay showed FGFR3 was the direct target of miR-1O0.FGFR3 was significantly downregulated by overexpressing miR-100 in pancreatic cancer cells (P ＜0.05),and FGFR3 knockdown by specific siRNA exerted the similar effect as miR-100 overexpression (P ＜ 0.05).Conclusions Our study identified a new miRNA regulator,miR-100,and clarified a novel mechanism of how miR-100 regulates cell proliferation in pancreatic cancer.The strategy of overexpressing the tumor suppressor miR-100 may provide a new therapeutic approach for treating patients with pancreatic cancer.