BACKGROUND:With the continuous advancement of medical technology,stem cell therapy has been used to treat a variety of diseases,including amyotrophic lateral sclerosis. OBJECTIVE:To review the research progress of stem cell therapy for amyotrophic lateral sclerosis,and prospect the development trend of this field. METHODS:PubMed,China National Knowledge Infrastructure(CNKI),and WanFang Data were searched for articles published from 1995 to 2024 using the key words"amyotrophic lateral sclerosis,mesenchymal stem cells,neural stem/progenitor cells,pluripotent stem cells."A total of more than 1 700 articles were retrieved,and 58 articles were finally included in this review. RESULTS AND CONCLUSION:Amyotrophic lateral sclerosis is a neurodegenerative disease that affects lower motor neurons in the brainstem and spinal cord and upper motor neurons in the motor cortex.The related research of stem cells in the treatment of amyotrophic lateral sclerosis has become a research hotspot.In this review,we summarize the application of different types of stem cells in amyotrophic lateral sclerosis research,including mesenchymal stem cells,neural stem progenitor cells,and induced pluripotent stem cells,and evaluate the key points of preclinical research such as stem cell source,cell volume,stem cell modification methods,and drug delivery routes,which lays the foundation for the future application of stem cell therapy.

OBJECTIVE To explore the effects of isorhamnetin on the development of gastric cancer through up-regulation of solute carrier family 25 member 25 antisense RNA 1（SLC25A25-AS1）. METHODS Using BALB/c nude mice as the subjects， the xenograft tumor model was established by subcutaneously inoculating human gastric cancer MKN28 cells into the axillary region. The effects of low and high doses of isorhamnetin （20 and 40 mg/kg） on the tumor volume and mass in nude mice were investigated. MKN28 cells were selected and divided into control group， isorhamnetin group （70 μmol/L， similarly hereinafter）， isorhamnetin+knocking down negative control group， isorhamnetin+knocking down SLC25A25-AS1 group， isorhamnetin+ overexpression negative control group and isorhamnetin+overexpressing SLC25A25-AS1 group. Effects of knocking down/ overexpressing SLC25A25-AS1 on viability， apoptosis， migration and invasion ability of isorhamnetin-treated cells were detected. After verifying the targeting relationships between microRNA-212-3p （miR-212-3p） and SLC25A25-AS1， as well as phosphatase and tensin homologue deleted on chromosome 10 （PTEN）， the effects of knocking down/overexpressing SLC25A25-AS1 on the expression of miR-212-3p， PTEN mRNA， and PTEN protein in isorhamnetin-treated cells were investigated. RESULTS Compared with the model control group， tumor volume and mass of nude mice in the isorhamnetin low-dose and high-dose groups were reduced significantly， and the isorhamnetin high-dose group was significantly lower than the isorhamnetin low-dose group （P＜0.05）. miR-212-3p had targeting relationships with SLC25A25-AS1 and PTEN. Compared with the control group， the cell viability （intervened for 24， 48 h）， migration number， invasion number and miR-212-3p expression of cells in the isorhamnetin group， isorhamnetin+knocking down negative control group and isorhamnetin+overexpressing negative control group were significantly reduced or decreased or down-regulated， while the apoptosis rate， mRNA and protein expressions of PTEN were significantly increased or up-regulated （P＜0.05）. Compared with isorhamnetin group and isorhamnetin+knocking down negative control group， the cell viability， migration number， invasion number and miR-212-3p expression of cells in the isorhamnetin+knocking down SLC25A25-AS1 group were significantly increased or up- regulated， while the apoptosis rate， mRNA and protein expressions of PTEN were significantly reduced or down-regulated （P＜ 0.05）. Compared with isorhamnetin group and isorhamnetin+overexpressing negative control group， the cell viability， migration number， invasion number and miR-212-3p expression of cells in isorhamnetin+overexpressing SLC25A25-AS1 group were significantly reduced or decreased or down-regulated， while the apoptosis rate， PTEN mRNA and protein expressions were significantly increased or up-regulated （P＜0.05）. CONCLUSIONS Isorhamnetin may inhibit the development of gastric cancer by up-regulating the expression of SLC25A25-AS1， down-regulating miR-212-3p， and up-regulating the expression of PTEN， which is a downstream target of miR-212-3p.

[Objective] To retrospectively analyse the serological characteristics of hemolytic transfusion reactions caused by Rh and Kidd antibodies, and to provide reference for safe, timely, and effective blood transfusion. [Methods] Two cases of patients with RhCcEe and Kidd blood type who experienced allogeneic transfusion at Dazhou Central Hospital were selected. A series of immunohematological tests were performed, including ABO, RhDCcEe and Kidd blood typing, unexpected antibody screening and identification, crossmatching, direct antiglobulin test, acid elution test, and capillary centrifugation to separate the patient's own red blood cells from donated red blood cells. [Results] Unexpected antibody screening, antibody identification, and direct antiglobulin test were positive in both patients. Case 1 had anti-Jk in the plasma, but no specific antibodies were found in the eluate. Case 2 had anti-c and E in the plasma, and anti-E was detected in the eluate. High-speed capillary centrifugation revealed corresponding antigen-positive erythrocytes at the distal end of the blood samples of both patients. [Conclusion] Case 1 received Kidd allogeneic red blood cells, and case 2 received RhCcEe allogeneic red blood cells, and both patients developed the corresponding unexpected antibodies, which led to the occurrence of immune haemolytic blood transfusion reaction.

Objective To explore the development and validation of a prediction model for severe communi-ty-acquired pneumonia in adults based on peripheral blood inflammatory indicators.Methods Venous blood samples of 204 community-acquired pneumonia in adults patients admitted to 7 hospitals in Chongqing area from April 2021 to August 2022 were collected to detect C-reactive protein(CRP),peripheral white blood cell count(WBC),neutrophil to lymphocyte ratio(NLR),cytokines,lymphocyte subgroups and neutrophil CD64 index.All of patients were divided into a training group and a validation group according to the time of admis-sion.Univariate and multivariate Logistic regression were used to analyze the data of the training group,the characteristic factors of severe progression for pneumonia were selected to construct the nomogram model,and the data of the validation group was used to verify the model.The receiver operating characteristic(ROC)curve,calibration curve and decision curve analysis(DCA)were used to evaluate the prediction ability of the model for severe community-acquired pneumonia in adults.Results Logistic regression analysis showed that age,CRP,WBC,interleukin(IL)-4/interferon gamma ratio and IL-6/IL-10 ratio were independent risk factors for severe community-acquired pneumonia in adults.The area under the ROC curve of the nomogram model in the training group and the validation group was 0.893 and 0.880,respectively.The calibration curve and DCA results shown that the model had a good prediction effect for severe community-acquired pneumonia in adults.Conclusion The inflammatory indicators included in this model are simple and easy to obtain clinically.This model with good differentiation and accuracy,it can be used as a practical tool to predict severe community-ac-quired pneumonia in adults,and has certain clinical application value.

OBJECTIVE:To systematically evaluate the efficacy of rehabilitation exoskeleton robots on the lower limb motor function of stroke patients using Meta-analysis and to compare the efficacy of different lower limb exoskeleton robots,so as to provide a theoretical basis for the scientific selection of suitable exoskeleton robots for patients with post-stroke lower limb motor dysfunction. METHODS:Computer searches of the Cochrane Library,PubMed,Web of Science,Embase,CNKI,VIP,and WanFang Data were conducted to collect randomized controlled clinical studies on exploring lower extremity rehabilitation exoskeleton robots to improve lower limb motor function in stroke patients published from database inception to November 2022.Two researchers conducted the literature search and screening.The quality of the included literature was evaluated using the Cochrane 5.1.0 risk of bias assessment tool and the Jadad scale.Meta-analysis was performed using RevMan 5.4 and Stata 17.0 software. RESULTS:(1)Finally 22 publications were included,involving 865 patients(n=436 in the test group and n=429 in the control group),and the Jadad score showed that all the included articles were of high quality.(2)Meta-analysis results showed that the exoskeleton robot significantly improved the Fugl-Meyer Assessment of Lower Extremity score(mean difference[MD]=2.63,95%confidence interval[CI]:1.87-3.38,P<0.05),Berg Balance Scale score(MD=3.62,95%CI:1.21-6.03,P<0.05),Timed Up and Go score(MD=-2.77,95%CI:-4.48 to-1.05,P<0.05)and step frequency score(MD=3.15,95%CI:1.57-4.72,P<0.05)in stroke patients compared with the control group.However,there was no significant improvement in the Functional Ambulation Category Scale score(MD=0.30,95%CI:-0.01 to 0.61,P>0.05)and 6-minute walk test score(MD=3.77,95%CI:-6.60 to 14.14,P>0.05).(3)Network Meta-analysis results showed that compared with the conventional rehabilitation therapy,both the level-walking exoskeleton(MD=10.23,95%CI:3.81-27.49,P<0.05)and the body-weight support exoskeleton(MD=33.66,95%CI:11.49-98.54,P<0.05)improved the Fugl-Meyer Assessment of Lower Extremity score.Compared with the conventional rehabilitation therapy,body-weight support exoskeleton significantly improved the Berg Balance Scale scores(MD=79.86,95%CI:2.34-2 725.99,P<0.05).In terms of Fugl-Meyer Assessment of Lower Extremity and Berg Balance Scale scores,the ranking results were body-weight support exoskeleton>level-walking exoskeleton>conventional rehabilitation therapy.Compared with the conventional rehabilitation therapy,level-walking exoskeleton significantly improved the Functional Ambulation Category Scale score(MD=1.38,95%CI:1.00-1.90,P<0.05)and body-weight support exoskeleton significantly improved the Timed Up and Go score(MD=0.07,95%CI:0.01-0.51,P<0.05).In terms of Functional Ambulation Category Scale and Timed Up and Go scores,the ranking results were level-walking exoskeleton>body-weight support exoskeleton>conventional rehabilitation therapy. CONCLUSION:Rehabilitation exoskeleton robots can improve balance,walking and activities of daily living in stroke patients,with body-weight support exoskeleton being more effective in improving lower limb motor function and balance and level walking exoskeleton being more effective in improving functional walking and transfer.

OBJECTIVE:To systematically evaluate the efficacy of transcranial direct current stimulation on the motor function of patients with Parkinson's disease,and to compare the efficacy of transcranial direct current stimulation at different targets on the motor function of patients with Parkinson's disease,so as to provide a theoretical basis for the target selection of transcranial direct current stimulation in clinical practice. METHODS:Cochrane Library,PubMed,Web of Science,CNKI,VIP,WanFang Data were retrieved for randomized controlled trials on the improvement of motor function in patients with Parkinson's disease by transcranial direct current stimulation published from the database inception to January 2023.The keywords were"Parkinson,transcranial direct current stimulation"in English and Chinese.The quality of the included studies was evaluated using the Cochrane 5.1.0 risk of bias assessment tool and the PEDro scale.Meta-analysis of outcome indicators was performed using RevMan 5.4 and Stata 17.0 software. RESULTS:Fifteen randomized controlled trials were finally included,and the PEDro scale showed that all were high-quality or very high-quality studies.Meta-analysis showed that transcranial direct current stimulation significantly improved Unified-Parkinson Disease Rating Scale part III score[mean difference(MD)=-2.49,95%confidence interval(CI):-4.42 to-0.55,P<0.05),step frequency score(MD=0.07,95%CI:0.03-0.11,P<0.05)and step speed score(MD=0.02,95%CI:0.00-0.05,P<0.05),but not for Berg Balance Scale scores(MD=2.57,95%CI:-0.74 to 5.87,P>0.05).Network Meta-analysis probability ranking:In terms of Unified-Parkinson Disease Rating Scale part III scores,the probability ranking results of target stimulation efficacy were dorsal lateral prefrontal cortex(52.4%)>primary motor cortex(45.8%)>central point of the brain(1.8%)>conventional rehabilitation(0%);in terms of gait frequency scores,the probability probability ranking results of target stimulation efficacy were cerebellum(50.1%)>central point of the brain(45.8%)>dorsal lateral prefrontal cortex(3.9%)>primary motor cortex(0.2%)>conventional rehabilitation(0%);in terms of gait speed scores,the probability ranking results of target stimulation efficacy were cerebellum(64.8%)>dorsal lateral prefrontal cortex(23.8%)>central point of the brain(9.4%)>primary motor cortex(1.7%)>conventional rehabilitation(0.4%);in terms of Berg Balance Scale scores,the probability ranking results of target stimulation efficacy were cerebellum(77.4%)>dorsal lateral prefrontal cortex(20.7%)>central point of the brain(0.7%)>conventional rehabilitation(0.2%). CONCLUSION:Transcranial direct current stimulation significantly improves motor function of patients with Parkinson's disease,with better motor coordination in the dorsolateral prefrontal cortex and better walking and balance in the cerebellum.

Objective:To construct an artificial intelligence (AI)-assisted system based on deep learning for corneal in vivo confocal microscopy (IVCM) image recognition and to evaluate its value in clinical applications. Methods:A diagnostic study was conducted.A total of 18 860 corneal images were collected from 331 subjects who underwent IVCM examination at Renmin Hospital of Wuhan University and Zhongnan Hospital of Wuhan University from May 2021 to September 2022.The collected images were used for model training and testing after being reviewed and classified by corneal experts.The model design included a low-quality image filtering model, a corneal image diagnosis model, and a 4-layer identification model for corneal epithelium, Bowman membrane, stroma, and endothelium, to initially determine normal and abnormal corneal images and corresponding corneal layers.A human-machine competition was conducted with another 360 database-independent IVCM images to compare the accuracy and time spent on image recognition by three senior ophthalmologists and the AI system.In addition, 8 trainees without IVCM training and with less than three years of clinical experience were selected to recognize the same 360 images without and with model assistance to analyze the effectiveness of model assistance.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2021-K148).Results:The accuracy of this diagnostic model in screening high-quality images was 0.954.Its overall accuracy in identifying normal/abnormal corneal images was 0.916 and 0.896 in the internal and external test sets, respectively.Its accuracy reached 0.983, 0.925 in the internal test sets and 0.988, 0.929 in the external test sets in identifying corneal layers of normal and abnormal images, respectively.In the human-machine competition, the overall recognition accuracy of the model was 0.878, which was similar to the average accuracy of the three senior physicians and was approximately 300 times faster than the experts in recognition speed.Trainees assisted by the system achieved an accuracy of 0.816±0.043 in identifying corneal layers of normal and abnormal images, which was significantly higher than 0.669±0.061 without model assistance ( t=6.304, P<0.001). Conclusions:A deep learning-based assistant system for corneal IVCM image recognition is successfully constructed.This system can discriminate normal/abnormal corneal images and diagnose the corresponding corneal layer of the images, which can improve the efficiency of clinical diagnosis and assist doctors in training and learning.

Objective To compare the survival outcomes of bladder cancer with non-pure urothelium(BCa with n-pU)and bladder cancer with pure urothelium(BCa with pU)treated with robot-assisted radical cystectomy(RARC).Methods Clinical data of BCa patients treated with RARC in Nanjing Drum Tower Hospital during Oct.2014 and Mar.2022 were retrospectively analyzed.The patients were divided into n-pU group and pU group.After the baseline differences between groups were balanced with propensity score matching(PSM),the overall survival(OS)and recurrence-free survival(RFS)curve were plotted using Kaplan-Meier method and compared using Log-rank test.Univariate and multivariate analysis were performed with Cox model to identify the influencing factors of prognosis.Based on the results,a secondary grouping was performed to compare the survival differences between subgroups and further investigate the prognostic factors.Results After PSM,there were 53 pairs of BCa patients.There were no significant differences in the baseline data between the pU and n-pU groups(P＜0.05).Regardless of T stage,there were no significant differences in OS and RFS between the two groups(P=0.217,P=0.109).Univariate Cox regression analysis showed that T stage(＞T2)was a significant risk factor of OS and RFS(P＜0.05).In the early pathological stage(≤T2),there were no significant differences in OS and RFS(P=0.565,P=0.344).In the advanced pathological stage(＞T2),the OS and RFS of n-pU were significantly worse than those of pU patients(P=0.025,P=0.034).Conclusion The prognosis of BCa patients with n-pU who received RARC is significantly correlated with pathological status.At＞T2 stage,n-pU patients have worse prognosis than pU patients in the same pathological status.

Objective To explore the blood group changes of two acute myeloid leukemia patients with suspected O type,and their relationship with the therapeutic effect.Methods Serological analysis of ABO blood group of patients was carried out by microcolumn gel method,tube method and absorption-elution test,ABO blood group genotyping was per-formed by microfluidic chip method.Exons E2 to E7 of ABO gene were amplified by PCR and sequenced by Sanger method.Results The forward typing of two cases were both O type,but the reverse typing were both A type.The absorption-elution test results all showed detection of antigen A.ABO gene phenotype of the two cases were both A,with genotyping results as A102/A102 and A102/O01,respectively.Sequencing results showed that SNP sites of ABO blood group were 467T/T,261G/delG and 467C/T,respectively.In one case,the intensity of anti-A agglutination reaction changed significantly from weak to strong with the progress of treatment.Conclusion For clinical samples of acute myeloid leukemia patients with ABO for-ward and reverse typing discrepancy and suspected O type,the result of reverse typing should be valued,and absorption-e-lution test should be performed to further confirm the ABO blood type combining the genetic test results,so as to develop ap-propriate blood transfusion strategies for patients.

ObjectiveTo explore the interaction between bruceoside B and gut microbiota and the inhibitory activity of its metabolites on human lung cancer A549 cells， and to explore the value of bruceoside B in the treatment of non-small cell lung cancer（NSCLC）. MethodBruceoside B was co-incubated with the human gut microbiota under anoxic conditions in vitro， and ultra high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to analyze the metabolic transformation products. Cell counting kit-8（CCK-8） assay was performed to determine the effects of bruceoside B and its metabolites on the proliferation of human lung cancer A549 cells and the half inhibitory concentration（IC₅₀） was calculated. Five healthy male rats were gavaged with bruceoside B（2 mg·kg^-1） for 7 days after adaptive feeding. The feces of rats were collected before and after administration. 16S rRNA sequencing was used to assess gut microbiota. ResultBruceoside B was mainly metabolized to brusatol by human gut microbiota， the IC₅₀ of bruceoside B and the conversion product to A549 cells were 1 755.50， 19.57 μmol·L^-1， respectively， and the conversion product had a better activity at inhibiting A549 cells proliferation than bruceoside B. Additionally， The results of intestinal flora analysis showed no significant differences in α diversity and β diversity of gut microbiota after administration. In terms of species abundance， at the phylum level， bruceoside B decreased the relative abundance of Actinobacteriota and Proteobacteria， increased the relative abundance of Firmicutes， Patescibacteria and Cyanobacteria. At the genus level， bruceoside B decreased the relative abundance of Staphylococcus， Aerococcus and Psychrobacter， increased the relative abundance of Romboutsia， Lactobacillus， Clostridium sensu stricto 1， Norank-f-norank-o-Clostridia-UCG-014， Turicibacter， Allobaculum and Candidatus Saccharimonas. The results of functional prediction showed that the gut microbiota functional compositions were relatively stable. ConclusionBruceoside B can be deglycosylated by intestinal flora and converted into brusatol， with a significant increase in antitumor activity. The administration of bruceoside B will not cause significant changes in the structure and function of the intestinal flora， resulting in intestinal microecological balance disorders， and the administration appears to be beneficial to the intestinal flora of NSCLC patients.