Background::Scleroderma is characterized by inflammation and fibrosis, predominantly occurring in the skin and extending to various parts of the body. The pathophysiology of scleroderma is multifaceted, with the current understanding including endothelial damage, inflammatory cell infiltration, and fibroblast activation in its progression. Nonetheless, the mechanism of cellular interactions and the precise spatial distribution of these cellular events within the fibrotic tissues remain elusive, highlighting a critical gap in our comprehensive understanding of scleroderma’s pathogenesis.Methods::In this study, we administered bleomycin intradermally to the dorsal skin of four individual murine models. Subsequently, skin tissues were harvested at predetermined intervals for comprehensive spatial transcriptomic analysis to determine the spatial dynamics influencing scleroderma pathogenesis. To validate the possible results from bioinformatic analysis, further in vitro and in vivo experiments were conducted. Results::Analysis of the spatial transcriptome revealed significant alterations in cell clusters during the progression of scleroderma. Gene Ontology analysis identified disruptions in lipid metabolism as the disease advanced. Pseudotime analysis provided evidence for a phenotypic transition from adipocytes to fibroblasts. In vitro studies demonstrated increased expression of Col1a1 and α-SMA as the disease progressed. These fibroblasts have been identified as key contributors to the increasing inflammation. Co-culturing TGF-β induced adipocytes with RAW264.7 cells resulted in overexpression of pro-inflammatory cytokines in the RAW264.7 cells. Both in vitro and in vivo experiments confirmed adipocyte loss and fibroblast formation, with transformed fibroblasts showing pronounced pro-inflammatory characteristics, highlighting their crucial role in the disease mechanism. Conclusions::Our study showed the spatial distribution and dynamic alterations of various cell types during scleroderma progression. Crucially, we identified the transformation of adipocytes into fibroblasts as a key factor promoting disease advancement. These emergent fibroblasts intensify inflammation, indicating that research on these cell clusters could reveal key scleroderma mechanisms and guide future therapies.

Objective:To discussed the regulation role of calcium/calmodulin dependent protein kinase(CaMK4)on γδT cells in the pathogenesis of lupus nephritis.Methods:The proportion of γδT cells in peripheral blood of LN patients and healthy con-trols were compared by flow cytometry.Magnetic bead was used to separation LN patients and healthy controls of gamma delta T cells in peripheral blood.RNA-SEQ analysis were performed in γδT cells of peripheral blood from LN patients and healthy controls.Real-time PCR and western blot were used to verify the expression level of differential gene CaMK4.The expression levels of CD80,CD86 and CD40L on γδT cells treated with CaMK4 inhibitor were detected by flow cytometry.ELISA was used to measure the level of IL-17 secreted by γδT cells after CaMK4 inhibitor treatment.Results:The proportion of γδT cells in the peripheral blood of LN patients was lower than that of healthy control group.A total of 28 differential genes related to LN were screened by RNA-SEQ sequencing,among which CaMK4 had significant by differences at molecular level and protein level.After γδT cells from LN patients were treated with CaMK4 inhibitor,KN-93,the expressions of surface costimulatory molecules CD80,CD86 and CD40L was decreased,and the level of IL-17 also was decreased.Conclusion:CaMK4 regulates γδT cells to participate in the pathogenesis of LN by IL-17 secretion and the expression of costimulatory molecules.Inhibition of CaMK4 may become a new target for the treatment of LN.

The aim of this study was to prepare a self-assembled nanoparticle monkeypox vaccine candidate and study its immune response characteristics,so as to provide reference test data for its vaccine design.The antigen protein A29L-SpyTag and the backbone protein Mi3-SpyCatcher were expressed and purified by prokaryotic system,and nanoparticles A29L-Mi3 were prepared by chemical assembly,then the antibody titers were determined by ELISA,the antibody neutralization was determined by plaque test,and the cytokine secretion of lymphocytes was determined by flow cytometry to describe the immune response characteristics.Data showed that A29L-Mi3 nanoparticles were successfully prepared,and the particles were uniformly distributed in hollow cages,with an average particle size of(29±0.19)nm.After the A29L-Mi3 nanoparticle vaccine candidate was combined with SP01 adjuvant,the neutralizing antibody titer was stronger than that of the A29L protein candidate,and the A29L-Mi3 nanoparticle vaccine candidate could obtain neutralizing antibodies with similar titers after two immunizations.The level of mouse T lymphocyte immune response activated by the A29L-Mi3 nanoparticle vaccine candidate was higher than that of the A29L protein vaccine candidate.In conclusion,A29L-Mi3 protein nanoparticles with uniform structure have successfully assembled in vitro,which has strong immunogenicity and improved neutralization ability after combination with SP01 adjuvant,thus provided reference data for the optimization of immune programs.In addition,the level of cellular immune response is higher than that of A29L protein alone,which provides a reference for the design and development of monkeypox vaccine.

【Objective】 To explore the timing of surgical drainage for ureteral calculi with upper urinary tract infection. 【Methods】 Clinical data of 117 cases of ureteral calculi with upper urinary tract infection treated in our hospital during Jan.2018 and Jan.2020 were retrospectively analyzed. According to different treatment methods, the patients were divided into surgical drainage group and non-surgical drainage group. The patients’ age, gender, side of calculi, peak body temperature, time of onset, white blood cell (WBC) count, C-reactive protein (CRP) and other clinical indicators were compared between the two groups. The cutoff value of surgical drainage was determined with receiver operator characteristic (ROC) curve. 【Results】 The patients’ age, peak body temperature, WBC count and CRP level were the influencing factors of surgical drainage (P<0.05). Regression analysis showed that CRP (P<0.001), age (P=0.003) and WBC count (P=0.014) were independent risk factors for surgical drainage. The area under the ROC curve (AUC) of CRP, age, and WBC count were 0.923, 0.601, and 0.796, respectively. The cutoff value of CRP was 29.87 mg/L (sensitivity 79.4%, specificity 90.0%). Logistic regression model showed CRP was a significant clinical predictor. 【Conclusion】 Ureteral calculi with upper urinary tract infection need to be diagnosed and treated in time. Positive anti-infection should be performed during emergency treatment, and surgical drainage could be selected according to the value of CRP.

Objective : To analyze the genome structure and genetic characteristics of IncpGRT1 plasmids from Pseud⁃ omonas , and elucidate its potential transmission risk . Methods : The genomic DNA of the clinical isolate 15420352 was extracted after purification and preservation of the strain , and then the whole genome was sequenced , and then the type of the plasmid was identified . Sequence annotation and comparison of the backbone region and the accessory modules were performed on all five same type sequenced plasmids , including one plasmid p420352 - strA in this study and four from GenBank . The plasmids were annotated by RAST , Plasmidfinder , Blast , ResFinder , and ISfinder. The ORFs of the plasmid were annotated and drug resistance genes were found . Results : All five plasmids were classified as new IncpGRT1 type plasmids . The IncpGRT1 backbone genes or gene loci were in all five plasmids , and they contained an auxiliary replicon besides the primary IncpGRT1 replicon . Five IncpGRT1 plasmids carried at least three different accessory modules , including the srp region , the msr region , and a Tn5053 family transposon . Three resistance genes strA , strB , and mer were obtained in these plasmids , which were involved in resistance to two categories of antibiotics and heavy metals . We also found that these plasmids carried at least one virulence gene msr and five key transporters srp , emrE , mod , phn , and lpt , which could improve the environmental adaptability of the strains . Conclusion The IncpGRT1 plasmids have become the important vector for the accumulation and spread of some drug resistance genes and virulence genes in Pseudomonas , and have improved the environmental adaptability of the strain.

Advanced hepatocellular carcinoma has a high degree of malignancy and poor prognosis. Studies showed that there is a close relationship between the progression of hepatocellular carcinoma and the immune status in tumor microenvironment. Adoptive cell therapy showed anti-tumor effects and improve immunosuppression by infusing patients with activated specific immune cells, which become a central issue in tumor therapy and shown promising effects in the treatment of various malignant tumors, indicating great application potential. Adoptive cell therapy based on neoantigen may become a new hot spot in the treatment of hepatocellular carcinoma, and their application, safety and effectiveness evaluation, efficacy prediction and assessment have become urgent issues to be solved. The purpose of this article is to introduce the progress related to adoptive cell therapy for advanced hepatocellular carcinoma and elaborate the problems that need to be solved in the future.

Patients undergoing coronary artery bypass grafting (CABG) belong to the very high-risk group of atherosclerotic cardiovascular disease. Although CABG gets advantages in relieving symptoms and improving long-term outcomes, a significant risk of cardiovascular adverse events after surgery still exists and standardized secondary prevention is needed. Lipid management plays a critical role as a secondary preventive strategy in CABG. However, lipid management of CABG patients in real clinical setting is inadequate, including lack of standardized lipid-lowering strategy, low goal attainment rate, as well as poor long-term medication adherence. In recent years, a series of clinical trials have provided a lot of groundbreaking new evidence for lipid management in patients with cardiovascular diseases which offers new strategies together with objectives of lipid-lowering and comprehensive management for patients undergoing CABG. This article reviews the strategy and research progress of lipid management after CABG, aiming to provide objective reference for clinical treatment.

Objective    To systematically review the models for predicting coronary artery disease (CAD) and demonstrate their predictive efficacy. Methods    PubMed, EMbase and China National Knowledge Internet were searched comprehensively by computer. We included studies which were designed to develop and validate predictive models of CAD. The studies published from inception to September 30, 2020 were searched. Two reviewers independently evaluated the studies according to the inclusion and exclusion criteria and extracted the baseline characteristics and metrics of model performance. Results    A total of 30 studies were identified, and 19 diagnostic predictive models were for CAD. Seventeen models had external validation group with area under curve (AUC)>0.7. The AUC for the external validation of the traditional models, including Diamond-Forrester model, updated Diamond-Forrester model, Duke Clinical Score, CAD consortium clinical score, ranged from 0.49 to 0.87. Conclusion    Most models have modest discriminative ability. The predictive efficacy of traditional models varies greatly among different populations.

Objective:To construct a prediction model for the prognosis of breast cancer patients with long non-coding RNA expression characteristics.Methods:To construct a long non-coding RNA(LncRNA) model for predicting the prognosis of breast cancer patients.Methods Analyzing LncRNA expression profiles and clinical characteristics of 1 081 breast cancer patients in the cancer genome atlas (TCGA) database.Performing differential expression analysis and univariate analysis on 112 paired breast cancer and normal breast tissues′ transcriptome sequencing data in the TCGA database, and screened for differentially expressed (DELncRNAs) that significantly correlated with the prognosis of BRCA (To reduce batch effects, sequencing data has been normalized using the DESeq function). One thousand eighty-one breast cancer patients were randomly divided into two groups: training set (541) and validation set (540). Performing Cox proportional hazard regression using DELncRNAs and establishing a multi-LncRNA prognosis model in the training set, followed by proportional hazards assumption test(PH assumption test). Patients were divided into high-risk and low-risk groups based on calculated risk score.Kaplan-Meier method was used for survival analysis, and 540 patients′ data were used for validation.To evaluate the prognostic value of the model in patients with squamous cell carcinoma of the lung and hepatocarcinoma in TCGA database.Gene Set Enrichment Analysis (GSEA) was used to analyze the specific mechanism of lncrna affecting the survival of patients.Results:There were 2815 differentially expressed genes screened by transcriptome sequencing, 91 of which were significantly related to the prognosis of breast cancer patients ( P<0.05). Based on the Cox regression analysis of 91 delncrna expression data from 541 breast cancer patients in training set, a Cox proportional risk regression model was constructed based on 5 LncRNA (training set AUC=0.746, validation set AUC=0.650): AC004551.1, MTOR-AS1, KCNAB1-AS2, FAM230G and LINC01283, and PH assumption test( P=0.388). K-M survival analysis showed that the survival time of high-risk group was significantly worse than that of low-risk group (median survival time: 7.049 and 12.21 years, HR 0.367, 95% CI0.228-0.597, P<0.001), and the survival time of high-risk group was significantly shorter than that of low-risk group (median survival time: 7.57 and 10.85 years, HR 0.412, 95% CI0.214-0.793, P<0.001). Similar prediction results were also obtained in other cancer species of TCGA: lung squamous cell carcinoma ( HR 0.604, 95% CI0.383-0.951, P=0.007) and liver cell carcinoma ( HR 0.551, 95% CI0.307-0.987, P=0.011). GSEA results suggested that the expression patterns of the above five LncRNA were related to the cell cycle regulation of tumor cells. Conclusion:The prognostic model constructed based on expression profile of AC004551.1, MTOR-AS1, KCNAB1-AS2, FAM230G and LINC01283 can be used to predict the prognosis of breast cancer patients, which is helpful to further guide clinical treatment.

Objective:To establish a predictive model that can predict the effectiveness and safety of endoscopic surgery for pediatric upper urinary tract calculi, and evaluate its diagnostic efficacy through internal validation.Methods:The data was selected from the prospective database of pediatric upper tract calculi constructed by Beijing Friendship Hospital Affiliated to Capital Medical University from June 2014 to April 2019. A total of 348 children were recruited in this investigation including 250 boys and 98 girls, with a median age of 3.0 years(Interquartile Range 1.4, 7.0 years). Totally 375 endoscopic surgeries were performed, with an overall stone free rate (SFR) of 88.0% (330/375) and complication rate (CR) of 23.2% (87/375). This research used univariate and multivariate logistic regression analysis to evaluate and screen the predictors of SFR and CR. The nomogram of SFR and CR was established by using the selected predictive factors. The differentiation degree of the model was evaluated by the area under the curve (AUC), the consistency between the prediction probability and the actual risk was evaluated by the calibration curve, and the clinical benefits of the model application were assessed by the decision curve.Results:The results of multivariate analysis demonstrated that stone burden, operation duration, intraoperative perfusion, stone location and operative options were the postoperative predictors of SFR and CR for children. Besides, BMI was also a predictor of postoperative CR. The AUC of the model (model 1: SFR; model 2: CR) based on the above predictive factors was 0.81 and 0.73, respectively. The predictive probability of the calibration curve showed that the model had a good consistency with the actual value. The decision curve showed that the application of model 1 to predict SFR had significant clinical benefits when the threshold value was greater than 20%, and the application of model 2 to predict the postoperative CR had a significant clinical benefit when the threshold was greater than 10%.Conclusions:Nomogram based on stone burden, operation time, intraoperative perfusion, stone location and operative options was validated internally with preferable predictive power on the effectiveness and safety of pediatric endoscopic surgery. This model can be used to predict the clinical effects of pediatric endoscopic lithotripsy. BMI was also an important factor for the safety of pediatric endoscopic procedures.