PURPOSE: To investigate the incidence and influencing factors of bone loss in patients with spinal cord injury (SCI). METHODS: A retrospective case-control study was conducted. Patients with SCI in our hospital from January 2019 to March 2023 were collected. According to the correlation between bone mineral density (BMD) at different sites, the patients were divided into the lumbar spine group and the hip joint group. According to the BMD value, the patients were divided into the normal bone mass group (t > -1.0 standard deviation) and the osteopenia group (t ≤ -1.0 standard deviation). The influencing factors accumulated as follows: gender, age, height, weight, cause of injury, injury segment, injury degree, time after injury, start time of rehabilitation, motor score, sensory score, spasticity, serum value of alkaline phosphatase, calcium, and phosphorus. The trend chart was drawn and the influencing factors were analyzed. SPSS 26.0 was used for statistical analysis. Correlation analysis was used to test the correlation between the BMD values of the lumbar spine and bilateral hips. Binary logistic regression analysis was used to explore the influencing factors of osteoporosis after SCI. p < 0.05 was considered statistically significant. RESULTS: The incidence of bone loss in patients with SCI was 66.3%. There was a low concordance between bone loss in the lumbar spine and the hip, and the hip was particularly susceptible to bone loss after SCI, with an upward trend in incidence (36% - 82%). In this study, patients with SCI were divided into the lumbar spine group (n = 100) and the hip group (n = 185) according to the BMD values of different sites. Then, the lumbar spine group was divided into the normal bone mass group (n = 53) and the osteopenia group (n = 47); the hip joint group was divided into the normal bone mass group (n = 83) and the osteopenia group (n = 102). Of these, lumbar bone loss after SCI is correlated with gender and weight (p = 0.032 and < 0.001, respectively), and hip bone loss is correlated with gender, height, weight, and time since injury (p < 0.001, p = 0.015, 0.009, and 0.012, respectively). CONCLUSIONS The incidence of bone loss after SCI was high, especially in the hip. The incidence and influencing factors of bone loss in the lumbar spine and hip were different. Patients with SCI who are male, low height, lightweight, and long time after injury were more likely to have bone loss.
Humans ; Spinal Cord Injuries/complications* ; Male ; Female ; Retrospective Studies ; Incidence ; Adult ; Bone Density ; Middle Aged ; Case-Control Studies ; Osteoporosis/etiology* ; Lumbar Vertebrae ; Bone Diseases, Metabolic/etiology* ; Aged ; Risk Factors

OBJECTIVE: To prepare clinical-grade human umbilical cord-derived mesenchymal stem cells (hUC-MSC) with high expression of hematopoietic supporting factors and evaluate their stem cell characteristics. METHODS: Fetal umbilical cord tissues were collected from healthy postpartum women during full-term cesarean section. Wharton's jelly was mechanically separated and hUC-MSCs were obtained by explant culture method and enzyme digestion method in an animal serum-free culture system with addition of human platelet lysate. The phenotypic characteristics of hUC-MSCs obtained by two methods were detected by flow cytometry. The differences in proliferation ability between the two groups of hUC-MSCs were identified through CCK-8 assay and colony forming unit-fibroblast (CFU-F) assay. The differences in multilineage differentiation potential between the two groups of hUC-MSCs were identified through induction of adipogenic, osteogenic, and chondrogenic differentiation. The mRNA expression levels of hematopoietic supporting factors such as SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in the two groups of hUC-MSCs were identified by real-time fluorescence quantiative PCR(RT-qPCR). RESULTS: The results of flow cytometry showed that hUC-MSCs obtained by the two methods both expressed high levels of CD73, CD90 and CD105, while lowly expressed CD31, CD45 and HLA-DR. The results of CCK-8 and CFU-F assay showed that the proliferation ability of hUC-MSCs obtained by explant culture method was better than those obtained by enzyme digestion method. The results of the triple lineage differentiation experiment showed that there was no significant difference in multilineage differentiation potential between the two grous of hUC-MSCs. The results of RT-qPCR showed that the mRNA expression levels of hematopoietic supporting factors SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in hUC-MSCs obtained by explant cultrue method were higher than those obtained by enzyme digestion method. CONCLUSION Clinical-grade hUC-MSCs with high expression levels of hematopoietic supporting factors were successfully cultured in an animal serum-free culture system.
Humans ; Mesenchymal Stem Cells/metabolism* ; Umbilical Cord/cytology* ; Cell Differentiation ; Female ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12/metabolism* ; Angiopoietin-1/metabolism* ; Vascular Cell Adhesion Molecule-1/metabolism* ; Stem Cell Factor/metabolism* ; Flow Cytometry ; Pregnancy

This study was aimed at developing an in vitro fluorescent substrate assay for the activity of leucyl aminopeptid-ase(LAP)from Echinococcus multilocularis and comparing it with the chemical chromogenic substrate enzyme activity assay.Through the establishment of reaction conditions for the fluorescent substrate-based in vitro enzyme activity assay,we com-pared the differences between the fluorescent substrate L-Leucine-7-amido-4-methylocoumarin(Leu-AMC)and the chemical chromogenic substrate L-Leucine-4-nitroanilide(Leu-pNA)through molecular docking,inhibition rates,and precision measures.Molecular docking revealed that the fluorescent substrate Leu-AMC had higher affinity for the protein than the chemical chromogenic substrate Leu-pNA.Through analysis of the effects of varying reaction conditions on fluorescence intensi-ty,we optimized the fluorescent substrate enzyme activity assay to demonstrate favorable performance at a reaction temperature of 37℃,a pH of 9.0,a protein concentration of 800 nmol/L,and a reaction duration of 60 minutes.Leu-AMC exhibited significant and distinct responses at a 5 μmol/L substrate concentration,under varying substrate conditions.The fluo-rescent substrate assay demonstrated more significant intergroup differences than the chemical chromogenic substrate assay when various inhibitors were added.This study established a fluorescence-based enzyme activity assay for leucyl aminopeptidase from Echinococcus multilocularis by using Leu-AMC as the substrate;this method demonstrated a more significant intergroup difference and sensitivity than the chemical chromogenic substrate assay.

Objective To analyze risk factors for poor postoperative wound healing in spinal tuberculo-sis patients and construct prediction models along with a risk scoring table using machine learning algorithms,providing references for early prevention and management.Methods Clinical data from 420 spinal tuberculo-sis patients treated at four tertiary hospitals in Guizhou Province between January 2017 and February 2024 were retrospectively analyzed.Risk factors were identified through univariate and multivariate analyses.Logis-tic regression,random forest,and support vector machine prediction models were constructed.Model perform-ance was evaluated using receiver operating characteristic(ROC)curves,precision,recall,accuracy,and F1-score.A risk prediction scoring table for poor postoperative wound healing was subsequently developed.Re-sults Among the 420 patients with spinal tuberculosis,132 experienced poor postoperative wound healing,with an incidence rate of 31.43%.Logistic regression analysis showed that BMI≤18.5 kg/m2,preoperative albumin≤30 g/L,combined surgical procedure,intraoperative blood loss>1 000 mL,direct bilirubin>8 μmol/L within 3 days after surgery,neutrophil count≤75×109/L within 3 days after surgery,and postopera-tive drainage volume>500 mL were risk factors for poor postoperative wound healing(P<0.05).In the comparison among the three risk prediction models constructed based on machine learning algorithms,the ran-dom forest model demonstrated the best predictive performance.A risk prediction scoring table was construc-ted based on the partial regression coefficients from the multivariate analysis,with a total score range from 0 to 11 points.A score>6 points indicated an increased risk of poor postoperative wound healing.The area un-der the ROC curve(AUC)for this risk prediction scoring table was 0.846(95%CI:0.769 to 0.923),indica-ting good predictive performance.Conclusion The random forest model based on machine learning algorithms and the risk prediction scoring table have certain predictive value for assessing the risk of poor postoperative wound healing in patients with spinal tuberculosis;they can assist healthcare professionals in early identifica-tion of high-risk patients.

This study was aimed atanalyzing the epidemic characteristics of dengue fever in Ruili City atthe China-Myanmar bor-der in late 2023,to provide evidence for local dengue fever prevention and control measures.Adult Aedes mosquitoes were collected from Ruili City with a backpack type mosquito sucking machine in October of 2023.Serum samples frompatients with suspected den-gue were collected in acutephase,in November of 2023.Detection ofdengue virus(DENV)nucleic acids in Aedesmosquitoes and acute phase serum samples from suspected dengue fever patients using reverse transcription-polymerase chain reaction,and nucleic acid positive samples were inoculated into Vero cells for viral culture.After three consecutive blind passage,samples with cytopathic effect(CPE)were collected for be sequencingand analysisof genetic and evolutionary information.Dengue case characteristics were analyzed through descriptive epidemiological methods.Among the 109 cases of dengue fever,the ratio of males to females was 1.27∶1.The youngest patient was 1 year old,the oldest patient was 84 years old,the age group of 20～59 years accounted for 73.39%,and the major-ity of occupations were freelancer(40.37%).A total of 827 female Aedes albopictus and 312 Aedes aegypti were collected,all of which tested negative for DENV nucleic acid.109 serum samples tested positive for DENV nucleic acid,including 49 DENV-1 and 60 DENV-2.Moreover,five DENV-1 and nine DENV-2 samples were obtainedthrough third-generation blind passaging with CPE.The E gene sequences of these five DENV-1 strains were detected,all were found to belong to DENV-1 genotype I,and had same evolu-tionary branch as the 2023 Guangzhou,China(PP563911),the 2019 Myanmar(MW793710),and 2019 Attapeu,Laos(MW559046).The nucleotide and amino acid sequence similarityamong the five DENV-1 genotype was 99.4%-99.9%and 99.8%-100.0%.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.5%-100.0%,99.4%-99.6%and 99.3%-99.5%,respec-tively,and amino acid similarity was 99.8%-100.0%.Nine DENV-2 E gene sequences were of Asian genotype I and belonged to the same evolutionary branch as the 2018 Myanmar(MW788982),2019 Hangzhou(OP684212)and 2019 Ruili(OQ928150).The nucleotide and amino acid similarities of the nine samples were 99.5%-100.0%and 99.8%-100.0%,respectively.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.7%-100.0%,99.3%-99.7%and 99.3%-99.7%,respectively,and the amino acid similarity was 99.8%-100%,99.8%-100.0%and 99.4%-99.6%,respectively.Two dengue vectors,Aedes albopictus and Aedes aegypti,were present in Ruili city,and the dengue outbreak was caused primarily by DENV-1 genotype I and DENV-2 Asian genotype I in later 2023.The sources of DENV-1 were probably the same as those of DENV-1 with Guangzhou(2023),and the sources of DENV-2 were probably from Myanmar.Dengue cases were found primarilyin the 20-59 year age group and freelancers,thus suggesting that relevant local departments should strengthen surveillance of dengue imported case and vector.

Objective:To establish a chain management model for blood glucose in critically ill patients after cardiac surgery and analyze its clinical effectiveness, and to provide a reference for related clinical nursing practices.Methods:A quasi-experimental study design was adopted. Using convenience sampling, 120 critically ill patients after cardiac surgery admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were selected as study subjects. They were divided into groups based on the time of admission to the intensive care unit after surgery. Sixty patients admitted from March to May 2024 were assigned to the control group and received conventional blood glucose management. Another 60 patients admitted from June to August 2024 were assigned to the observation group and received the chain management model for blood glucose control in addition to the conventional approach. Blood glucose levels and quality indicators of blood glucose management (including maximum blood glucose fluctuation, time in target glucose range, duration of insulin use, incidence of hypoglycemia, etc.) were compared between the two groups.Results:The control group included 27 males and 33 females, with an age of 63.00(59.00, 69.25) years; the observation group included 28 males and 32 females, with an age of 66.00(60.00, 70.00) years. The blood glucose levels of the observation group on postoperative days 2, 3, 4 were 9.10(8.68, 9.90), 8.90(8.40, 10.00), 8.75(7.38, 9.03) mmol/L, respectively, which were lower than those of the control group [10.30(9.80, 11.00), 9.95(9.40, 11.05), 9.30(8.10, 10.02) mmol/L], with a statistically significant difference ( Z=-5.85, -4.95, -3.50, all P<0.05). The maximum blood glucose fluctuation in the observation group was (4.09 ± 2.45) mmol/L, lower than that of the control group [(5.19 ± 2.47) mmol/L], with a statistically significant difference ( t=2.46, P<0.05). The time in the target glucose range was 67.00(60.00, 75.00)% in the observation group, higher than that of the control group 52.00(45.00, 60.00)%, with a statistically significant difference ( Z=-6.57, P<0.05). The duration of insulin use was 6.00(5.00, 7.00) h in the observation group, shorter than that of the control group [13.00(9.75, 15.32) h], with a statistically significant difference ( Z=-8.68, P<0.05). The incidence of hypoglycemia was 3.33%(2/60) in the observation group and 15.00%(9/60) in the control group, with a statistically significant difference ( χ2=4.90, P<0.05). The mechanical ventilation time, ICU stay, and total hospital stay in the observation group were 42.00(37.00, 89.25) h, 6.00(5.00, 7.00) d, and 12.00(11.75, 13.00) d, respectively, which were shorter than those of the control group [96.00(86.25, 98.00) h, 7.00(7.00, 10.00) d, and 13.00(11.75, 15.00) d], with a statistically significant difference ( Z=8.67, 17.57, 4.73, all P<0.05). Conclusions:The implemented chain management model for blood glucose control meets the comprehensive requirements of blood glucose management. It not only reduces blood glucose fluctuations and decreases the incidence of hypoglycemia but also effectively improves the quality of blood glucose management in critically ill patients after cardiac surgery, enhances the safety of blood glucose control, and promotes patient recovery.

This study was aimed at developing an in vitro fluorescent substrate assay for the activity of leucyl aminopeptid-ase(LAP)from Echinococcus multilocularis and comparing it with the chemical chromogenic substrate enzyme activity assay.Through the establishment of reaction conditions for the fluorescent substrate-based in vitro enzyme activity assay,we com-pared the differences between the fluorescent substrate L-Leucine-7-amido-4-methylocoumarin(Leu-AMC)and the chemical chromogenic substrate L-Leucine-4-nitroanilide(Leu-pNA)through molecular docking,inhibition rates,and precision measures.Molecular docking revealed that the fluorescent substrate Leu-AMC had higher affinity for the protein than the chemical chromogenic substrate Leu-pNA.Through analysis of the effects of varying reaction conditions on fluorescence intensi-ty,we optimized the fluorescent substrate enzyme activity assay to demonstrate favorable performance at a reaction temperature of 37℃,a pH of 9.0,a protein concentration of 800 nmol/L,and a reaction duration of 60 minutes.Leu-AMC exhibited significant and distinct responses at a 5 μmol/L substrate concentration,under varying substrate conditions.The fluo-rescent substrate assay demonstrated more significant intergroup differences than the chemical chromogenic substrate assay when various inhibitors were added.This study established a fluorescence-based enzyme activity assay for leucyl aminopeptidase from Echinococcus multilocularis by using Leu-AMC as the substrate;this method demonstrated a more significant intergroup difference and sensitivity than the chemical chromogenic substrate assay.

This study was aimed atanalyzing the epidemic characteristics of dengue fever in Ruili City atthe China-Myanmar bor-der in late 2023,to provide evidence for local dengue fever prevention and control measures.Adult Aedes mosquitoes were collected from Ruili City with a backpack type mosquito sucking machine in October of 2023.Serum samples frompatients with suspected den-gue were collected in acutephase,in November of 2023.Detection ofdengue virus(DENV)nucleic acids in Aedesmosquitoes and acute phase serum samples from suspected dengue fever patients using reverse transcription-polymerase chain reaction,and nucleic acid positive samples were inoculated into Vero cells for viral culture.After three consecutive blind passage,samples with cytopathic effect(CPE)were collected for be sequencingand analysisof genetic and evolutionary information.Dengue case characteristics were analyzed through descriptive epidemiological methods.Among the 109 cases of dengue fever,the ratio of males to females was 1.27∶1.The youngest patient was 1 year old,the oldest patient was 84 years old,the age group of 20～59 years accounted for 73.39%,and the major-ity of occupations were freelancer(40.37%).A total of 827 female Aedes albopictus and 312 Aedes aegypti were collected,all of which tested negative for DENV nucleic acid.109 serum samples tested positive for DENV nucleic acid,including 49 DENV-1 and 60 DENV-2.Moreover,five DENV-1 and nine DENV-2 samples were obtainedthrough third-generation blind passaging with CPE.The E gene sequences of these five DENV-1 strains were detected,all were found to belong to DENV-1 genotype I,and had same evolu-tionary branch as the 2023 Guangzhou,China(PP563911),the 2019 Myanmar(MW793710),and 2019 Attapeu,Laos(MW559046).The nucleotide and amino acid sequence similarityamong the five DENV-1 genotype was 99.4%-99.9%and 99.8%-100.0%.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.5%-100.0%,99.4%-99.6%and 99.3%-99.5%,respec-tively,and amino acid similarity was 99.8%-100.0%.Nine DENV-2 E gene sequences were of Asian genotype I and belonged to the same evolutionary branch as the 2018 Myanmar(MW788982),2019 Hangzhou(OP684212)and 2019 Ruili(OQ928150).The nucleotide and amino acid similarities of the nine samples were 99.5%-100.0%and 99.8%-100.0%,respectively.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.7%-100.0%,99.3%-99.7%and 99.3%-99.7%,respectively,and the amino acid similarity was 99.8%-100%,99.8%-100.0%and 99.4%-99.6%,respectively.Two dengue vectors,Aedes albopictus and Aedes aegypti,were present in Ruili city,and the dengue outbreak was caused primarily by DENV-1 genotype I and DENV-2 Asian genotype I in later 2023.The sources of DENV-1 were probably the same as those of DENV-1 with Guangzhou(2023),and the sources of DENV-2 were probably from Myanmar.Dengue cases were found primarilyin the 20-59 year age group and freelancers,thus suggesting that relevant local departments should strengthen surveillance of dengue imported case and vector.

Objective:To establish a chain management model for blood glucose in critically ill patients after cardiac surgery and analyze its clinical effectiveness, and to provide a reference for related clinical nursing practices.Methods:A quasi-experimental study design was adopted. Using convenience sampling, 120 critically ill patients after cardiac surgery admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were selected as study subjects. They were divided into groups based on the time of admission to the intensive care unit after surgery. Sixty patients admitted from March to May 2024 were assigned to the control group and received conventional blood glucose management. Another 60 patients admitted from June to August 2024 were assigned to the observation group and received the chain management model for blood glucose control in addition to the conventional approach. Blood glucose levels and quality indicators of blood glucose management (including maximum blood glucose fluctuation, time in target glucose range, duration of insulin use, incidence of hypoglycemia, etc.) were compared between the two groups.Results:The control group included 27 males and 33 females, with an age of 63.00(59.00, 69.25) years; the observation group included 28 males and 32 females, with an age of 66.00(60.00, 70.00) years. The blood glucose levels of the observation group on postoperative days 2, 3, 4 were 9.10(8.68, 9.90), 8.90(8.40, 10.00), 8.75(7.38, 9.03) mmol/L, respectively, which were lower than those of the control group [10.30(9.80, 11.00), 9.95(9.40, 11.05), 9.30(8.10, 10.02) mmol/L], with a statistically significant difference ( Z=-5.85, -4.95, -3.50, all P<0.05). The maximum blood glucose fluctuation in the observation group was (4.09 ± 2.45) mmol/L, lower than that of the control group [(5.19 ± 2.47) mmol/L], with a statistically significant difference ( t=2.46, P<0.05). The time in the target glucose range was 67.00(60.00, 75.00)% in the observation group, higher than that of the control group 52.00(45.00, 60.00)%, with a statistically significant difference ( Z=-6.57, P<0.05). The duration of insulin use was 6.00(5.00, 7.00) h in the observation group, shorter than that of the control group [13.00(9.75, 15.32) h], with a statistically significant difference ( Z=-8.68, P<0.05). The incidence of hypoglycemia was 3.33%(2/60) in the observation group and 15.00%(9/60) in the control group, with a statistically significant difference ( χ2=4.90, P<0.05). The mechanical ventilation time, ICU stay, and total hospital stay in the observation group were 42.00(37.00, 89.25) h, 6.00(5.00, 7.00) d, and 12.00(11.75, 13.00) d, respectively, which were shorter than those of the control group [96.00(86.25, 98.00) h, 7.00(7.00, 10.00) d, and 13.00(11.75, 15.00) d], with a statistically significant difference ( Z=8.67, 17.57, 4.73, all P<0.05). Conclusions:The implemented chain management model for blood glucose control meets the comprehensive requirements of blood glucose management. It not only reduces blood glucose fluctuations and decreases the incidence of hypoglycemia but also effectively improves the quality of blood glucose management in critically ill patients after cardiac surgery, enhances the safety of blood glucose control, and promotes patient recovery.

Objective:To investigate the effects and underlying mechanism of ionizing radiation on the adipogenic of mesenchymal stem cells(MSCs).Methods:Mouse MSCs were cultured in vitro and treated with 2 Gy and 6 Gy radiation with 60Co,and the radiation dose rate was 0.98 Gy/min.Bulk RNA-seq was performed on control and irradiated MSCs.The changes of adipogenic differentiation and oxidative stress pathways of MSC were revealed by bioinformatics analysis.Oil Red O staining was used to detect the adipogenic differentiation ability of MSCs in vitro,and real-time fluorescence quantitative PCR(qPCR)was used to detect the expression differences of key regulatory factors Cebpa,Lpl and Pparg after radiation treatment.At the same time,qPCR and Western blot were used to detect the effect of inhibition of Nrf2,a key factor of antioxidant stress pathway,on the expression of key regulatory factors of adipogenesis.Moreover,the species conservation of the irradiation response of human bone marrow MSCs and mouse MSC was determined by qPCR.Results:Bulk RNA-seq suggested that ionizing radiation promotes adipogenic differentiation of MSCs and up-regulation of oxidative stress-related genes and pathways.The results of Oil Red O staining and qPCR showed that ionizing radiation promoted the adipogenesis of MSCs,with high expression of Cebpa,Lpl and Pparg,as well as oxidative stress-related gene Nrf2.Nrf2 pathway inhibitors could further enhance the adipogenesis of MSCs in bone marrow after radiation.Notably,the similar regulation of oxidative pathways and enhanced adipogenesis post irradiation were observed in human bone marrow MSCs.In addition,irradiation exposure led to up-regulated mRNA expression of interleukin-6 and down-regulated mRNA expression of colony stimulating factor 2 in human bone marrow MSCs.Conclusion:Ionizing radiation promotes adipogenesis of MSCs in mice,and oxidative stress pathway participates in this effect,blocking Nrf2 further promotes the adipogenesis of MSCs.Additionally,irradiation activates oxidative pathways and promotes adipogenic differentiation of human bone marrow MSCs.