Objective:To investigate the efficacy of hybrid fixation with tibial prosthesis in primary total knee arthroplasty (TKA) in patients with osteoporosis or osteopenia.Methods:A total of 39 patients (42 knees) with knee osteoarthritis who underwent primary TKA with hybrid fixation with tibial prosthesis in the First Affiliated Hospital of Zhejiang University from November 2022 to June 2024 were retrospectively analyzed. There were 12 males and 27 females with an average age of 68.23±7.02 years (range, 58-89 years). The mean body mass index was 26.15±4.63 kg/m 2 (range, 19.48-39.03 kg/m 2), and the mean bone mineral density T score was -2.38±0.78 (from -4.2 to -1.2). The left knee was 19, and the right knee was 23. The visual analogue scale (VAS) and Knee Society score (KSS) were used to evaluate the knee joint pain and function, and the knee flexion range of motion was measured. Radiographic evaluation included hip-knee-ankle angle, lateral distal femoral angle, and posterior slope of tibial plateau. Results:All patients successfully completed the operation and were followed up for 19.8±5.1 months (range, 4.9-23.6 months). All patients were treated with hybrid fixation with tibial prosthesis assisted by 3D printed porous titanium alloy sleeve. The operation time was 73.8±10.8 min (range, 54-98 min). Thirty-four of 39 patients received standard anti-osteoporosis therapy before operation. The VAS score at the last follow-up was 0(0, 1), which was lower than that before operation 7(6,8), and the difference was statistically significant ( U<0.001, Ρ<0.001). The KSS clinical score at the last follow-up was 76.67±10.04, which was higher than that before operation (50.95±7.35), and the difference was statistically significant ( t=12.030, Ρ<0.001). The knee flexion range of motion at last follow-up was 97.26°±12.94°, showing no significant difference when compared with preoperative 94.52°±16.52° ( t=0.846, Ρ=0.400). The hip-knee-ankle angle at last follow-up was 176.2°±2.54°, which was significantly higher than that before operation 173.5°±5.25° ( t=3.328, Ρ=0.002). The distal lateral femoral and posterior tibial inclination angles at the final follow-up were 89.58°±1.62° and 2.85°±0.91°, respectively, compared with 90.71°±4.01° and 3.05°±1.25° preoperatively, with no statistically significant differences ( t=1.556, Ρ=0.127; t=0.923, Ρ=0.359). No prosthesis loosening, dislocation, periprosthetic infection, deep vein thrombosis and other complications occurred. Conclusion:Hybrid fixation with tibial prosthesis in primary TKA for patients with osteoporosis or osteopenia has a satisfactory clinical effect, which can relieve knee pain and improve joint function.

BACKGROUND:Overexpression of the nuclear factor I-C gene in vitro promotes the differentiation of human stem cells from apical papilla,as does the activation of the Wnt/β-catenin signaling pathway.Moreover,nuclear factor I-C regulates the Wnt/β-catenin pathway in mesenchymal stem cells.However,whether nuclear factor I-C can affect cell differentiation by activating the Wnt/β-catenin pathway inhuman stem cells from apical papilla has not been reported.OBJECTIVE:To investigate the role of nuclear factor I-C in the Wnt/β-catenin signaling pathway in regulating the differentiation of human stem cells from apical papilla.METHODS:H uman stem cells from apical papilla were cultured by the slide-covered tissue block method and lentiviral transfection overexpressing the nuclear factor I-C gene.(1)A control group,an empty viral vector group,and an overexpressed nuclear factor I-C gene group were set up.The expression ofβ-Catenin,LRP5,and TCF7L2 was detected by Western blotting.(2)The control group,empty viral vector group,overexpressed nuclear factor I-C gene group,and overexpressed nuclear factor I-C gene+DKK-1(Wnt pathway inhibitor)group were set up.Alkaline phosphatase staining and activity quantification were performed after 7 days of osteogenic induction.qPCR and Western blotting were performed to detect the expression of Runt-related transcription factor 2,dentin salivary phosphoprotein,osteocalcin mRNA,and protein after 14 days of osteogenic induction.Alizarin Red staining was used to observe the formation of mineralized nodules.RESULTS AND CONCLUSION:(1)Compared with the control and empty viral vector groups,the expression of Wnt/β-Catenin pathway-related proteins β-Catenin,LRP5,and TCF7L2 inhuman apical dentin papilla stem cells was significantly increased in the overexpressed nuclear factor I-C gene group(P＜0.01).(2)Compared with the control and empty viral vector groups,the expression of alkaline phosphatase and osteocalcin in human apical dentin papilla stem cells was significantly increased(P＜0.01);the expression levels of Runt-related transcription factor 2,dentin salivary phosphoprotein,osteocalcin mRNA and protein were significantly higher(P＜0.01),and the number of mineralized nodules was significantly increased(P＜0.01)in the overexpressed nuclear factor I-C gene group.(3)Compared with the overexpressed nuclear factor I-C gene group,the alkaline phosphatase activity and the expression of Runt-related transcription factor 2,dentin salivary phosphoprotein,osteocalcin mRNA and protein expression levels were significantly down-regulated(P＜0.05),and the number of mineralized nodules was significantly reduced(P＜0.05)in human stem cells from apical papilla of the overexpressed nuclear factor I-C gene+DKK-1 group.The results show that nuclear factor I-C can activate the Wnt/β-catenin signaling pathway in human stem cells from apical papilla and mediate the osteogenic/odontogenic differentiation of human stem cells from apical papilla.

Objective To explore the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponin,and the UGT gene in Dipsacus asper Wall.ex Henry has been analyzed by the identification of whole genome,genome and prokaryotic expression.Methods The laboratory self-tested sequenced protein sequence files of the Dipsacus asper Wall.ex Henry genome were used.To validate the conserved domains of the sequence of the Dipsacus asper Wall.ex Henry UGT gene,BLASTP and hmmsearch were utilized.Prot-Param,SOMPA,MAGA7.0,Tbtools and other tools were used to investigate the protein physicochemical properties,protein structure,and covariance analysis of the Dipsacus asper Wall.ex Henry UGT gene family,and using the joint analysis of transcriptomic data and metabolomics data,two glycosyltransferases that might be related to triterpene saponin biosynthesis were screened,and expression vectors were constructed for prokaryotic expression.Results 44 Dipsacus asper Wall.ex Henry UGT genes were identified from the Dipsacus asper Wall.ex Henry genome.The length of Dipsacus asper Wall.ex Henry UGT proteins ranged from 49 to 1083 amino acids,with an average molecular weight of 24.86 kDa and an isoelectric point of 4.31-8.59.Dipsacus asper Wall.ex Henry UGT gene family was distributed on eight chromosomes.The phylogenetic tree constructed from the sequences of Dipsacus asper Wall.ex Henry,Arabidopsis thaliana and identified UGTs showed that glycosyltransferase gene families in Dipsacus asper Wall.ex Henry were mainly in the UGT73,UGT81,UGT85,and UGT80 families.Cis-acting element analysis showed that light-responsive elements were the most prevalent elements in the promoter regions of UGT gene family members.Two glycosyltransferases potentially related to triterpene saponin biosynthesis were screened using combined transcriptomics and metabolomics analysis,and were successfully expressed in prokaryotic form.Conclusion In this study,two candidate genes related to the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponins were jointly screened for prokaryotic expression using multi-omics,and were subjected to prokaryotic expression for further validation of the function of the genes.

Objective To investigate the changes of serum levels of soluble scavenger receptor 163 (sCD163),angiopoietin-like protein 3 (ANGPTL3) before and after intravenous thrombolysis in patients with acute cerebral infarction (ACI) and their correlation with prognosis. Methods A total of 60 ACI patients accepted by Cangzhou Central Hospital from June 2021 to June 2022 were collected as the ACI group,and another 60 healthy individuals were regarded as the control group. According to the National Institutes of Health Stroke Scale (NIHSS) score after admission,60 patients were divided into mild group (n=10),moderate group (n=38) and severe group (n=12).According to the scores on the modified Rankin scale 90 days after thrombolysis,patients were separated into a good prognosis group (n=42) and a poor prognosis group (n=18). The serum levels of sCD163 and ANGPTL3 were detected using enzyme linked immunosorbent assay (ELISA),and receiver operating characteristic (ROC) curve was applied to analyze the predictive value of serum sCD163 and ANGPTL3 levels for the prognosis of ACI patients after intravenous thrombolysis therapy. Results Compared with the control group,the levels of serum sCD163 (687.55±86.43 ng/ml vs 411.07±58.24 ng/ml) and ANGPTL3 (60.28±10.55 mg/L vs 25.34±5.93 mg/L) in ACI group were significantly increased,and the differences were significant (t=20.549,22.363,all P<0.05). The levels of serum sCD163 (551.65±69.66 ng/ml,668.92±81.12 ng/ml,859.79±117.24 ng/ml) and ANGPTL3 (44.52±8.12 mg/L,58.67±10.37 mg/L,75.34±13.12 mg/L) in mild,moderate and severe groups were gradually increased,and the differences were significant (F=36.011,23.007,all P<0.05). Compared with the good prognosis group,the proportion of time from onset to thrombolysis≥ 3 h,the proportion of NIHSS score>10 at admission,and the serum sCD163 and ANGPTL3 levels before and after thrombolysis were significantly increased in the poor prognosis group,and the differences were statistically significant (t/x2=5.644,4.775,8.982,10.866,10.293,9.702,all P<0.05). ROC results showed that the area under the curves(95％ confidence intervals)[AUC(95％CI)]of serum sCD163 and ANGPTL3 level alone in predicting the prognosis of ACI patients were 0.830 (0.711～0.915) and 0.783 (0.658～0.879),and their sensitivity and specificity were 72.22％ and 85.71％,77.78％ and 85.71％,respectively. The AUC(95％CI)of combined prediction of serum sCD163 and ANGPTL3 in predicting the prognosis of ACI patients[0.950(0.861～0.990)]was obviously greater than the AUC predicted by sCD163 and ANGPTL3 alone (Z=2.378,2.109,P=0.017,0.035). Conclusion sCD163 and ANGPTL3 levels are elevated in the serum of ACI patients,and are related to their severity and prognosis.

OBJECTIVES: To analyze the disease burden and inequalities of lower extremity peripheral artery disease (LEPAD) among people aged 40 and above in the Belt and Road partner countries from 1990 to 2021. METHODS: Data were retrieved from the Global Burden of Disease 2021 database. The age-standardized prevalence rates, mortality rates, and the annual rate of years lived with disability (YLDs) of LEPAD were analyzed. Trends were measured using the estimated annual percentage change (EAPC), and the slope index of inequality (SII) and concentration index were used to quantify the absolute and relative inequalities. RESULTS: In 2021, the age-standardized prevalence and mortality rates of LEPAD were 3168.26/10⁵ and 3.09/10⁵, increasing by 4.30% and 19.31% compared to 1990, while YLDs rates decreased by 4.00%. Females had higher age-standardized prevalence and YLDs rates, while males had higher mortality rates. The EAPC for prevalence rates was slightly higher in males (0.22%) than in females (0.17%); while the EAPC of age-standardized mortality rate was 2.02% for females, compared to 1.45% for males. From 1990 to 2021, the age-standardized YLDs rates decreased from 16.23/10⁵ to 15.58/10⁵, with a faster decline in females (－0.12%) than in males (－0.06%). LEPAD prevalence varied across countries, with higher burden in Europe and faster growth in Gulf states. Higher socio-demographic index countries had higher prevalence. Inequity improved, with the SII at 52.90/10⁵ and concentration index at 0.038 in 2021. Gender disparities persisted, with concentration index increased to 0.058 in females and reduced to －0.026 in males. CONCLUSIONS LEPAD prevalence and mortality among people aged 40 and above in the Belt and Road partner countries increased, while YLDs rates decreased from 1990 to 2021. Significant differences among people exist depending on gender and country, highlighting the need for enhanced screening, health education, and shared public health strategies across the Belt and Road partner countries.
Humans ; Peripheral Arterial Disease/mortality* ; Male ; Female ; Middle Aged ; Adult ; Aged ; Prevalence ; Lower Extremity/blood supply* ; Global Burden of Disease ; Cost of Illness

Objective: To summarize strategies for improving childhood hypertension, so as to provide evidencebased recommendations for poliymaking and practice childhood hypertension management in China. Methods: From March to April 2024, child health stakeholders from five districts in Hangzhou were selected using a combination of stratified and convenience sampling methods. Data were analyzed using a groundedtheory approach. During the indepth interview phase, six policymakers were interviewed. Focus group discussions were conducted with school administrators, healthcare providers, and parents, comprising a total of 62 participants. Results: Through threelevel coding, 116 initial categories were identified(e.g., "trend of younger age" "difficulty in behavior change"), 35 main categories (e.g., "higher incidence compared to the past" "caused by comprehensive influencing factors"), and 12 core categories (e.g., "epidemic status" "influencing factors"). Finally, the cognitive status, problem analysis, and management strategies of children hypertension were constructed. Conclusion Effective prevention and control of childhood hypertension requires coordinated efforts among governments, schools, families, and society to establish a comprehensive management system, with dynamic monitoring and evaluation to optimize policy implementation.

Objective:To study the characteristics of soil microbial composition in sheep sheds in agricultural and pastoral areas, and explore potential zoonotic pathogen species.Methods:Using cross-sectional survey method, soil samples from sheep sheds in the agricultural and pastoral areas of Darhan-Muminggan Joint County, Baotou City, Inner Mongolia Autonomous Region were collected from August to October 2023. Genomic DNA was extracted, and metagenomic second-generation sequencing was conducted. Microbial species annotation was carried out by Kaiju method, and the soil microbial composition and zoonotic pathogen species of sheep sheds in agricultural and pastoral areas were analyzed.Results:A total of 10 and 5 soil samples were collected from sheep sheds in agricultural and pastoral areas, respectively. The results of β diversity analysis showed that there was a significant separation trend in soil samples from sheep sheds in agricultural and pastoral areas, and the interpretation of principal co-ordinates analysis (PCoA) 1 and PCoA2 were 27.8% and 17.4%, respectively. There was no significant difference in the microbial composition of soil samples from sheep sheds in agricultural and pastoral areas ( R = 0.09, P = 0.242). At the phylum level, the dominant bacterial phyla were Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. At the genus level, the dominant bacterial genera were Corynebacterium, Luteimonas, Atopostipes, and Salinicoccus. A variety of zoonotic pathogens were detected, including Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani in 15 soil samples. Conclusion:The soil microorganisms in sheep sheds in agricultural and pastoral areas are diverse, and zoonotic pathogens such as Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani are detected.

Objective:To investigate the efficacy of hybrid fixation with tibial prosthesis in primary total knee arthroplasty (TKA) in patients with osteoporosis or osteopenia.Methods:A total of 39 patients (42 knees) with knee osteoarthritis who underwent primary TKA with hybrid fixation with tibial prosthesis in the First Affiliated Hospital of Zhejiang University from November 2022 to June 2024 were retrospectively analyzed. There were 12 males and 27 females with an average age of 68.23±7.02 years (range, 58-89 years). The mean body mass index was 26.15±4.63 kg/m 2 (range, 19.48-39.03 kg/m 2), and the mean bone mineral density T score was -2.38±0.78 (from -4.2 to -1.2). The left knee was 19, and the right knee was 23. The visual analogue scale (VAS) and Knee Society score (KSS) were used to evaluate the knee joint pain and function, and the knee flexion range of motion was measured. Radiographic evaluation included hip-knee-ankle angle, lateral distal femoral angle, and posterior slope of tibial plateau. Results:All patients successfully completed the operation and were followed up for 19.8±5.1 months (range, 4.9-23.6 months). All patients were treated with hybrid fixation with tibial prosthesis assisted by 3D printed porous titanium alloy sleeve. The operation time was 73.8±10.8 min (range, 54-98 min). Thirty-four of 39 patients received standard anti-osteoporosis therapy before operation. The VAS score at the last follow-up was 0(0, 1), which was lower than that before operation 7(6,8), and the difference was statistically significant ( U<0.001, Ρ<0.001). The KSS clinical score at the last follow-up was 76.67±10.04, which was higher than that before operation (50.95±7.35), and the difference was statistically significant ( t=12.030, Ρ<0.001). The knee flexion range of motion at last follow-up was 97.26°±12.94°, showing no significant difference when compared with preoperative 94.52°±16.52° ( t=0.846, Ρ=0.400). The hip-knee-ankle angle at last follow-up was 176.2°±2.54°, which was significantly higher than that before operation 173.5°±5.25° ( t=3.328, Ρ=0.002). The distal lateral femoral and posterior tibial inclination angles at the final follow-up were 89.58°±1.62° and 2.85°±0.91°, respectively, compared with 90.71°±4.01° and 3.05°±1.25° preoperatively, with no statistically significant differences ( t=1.556, Ρ=0.127; t=0.923, Ρ=0.359). No prosthesis loosening, dislocation, periprosthetic infection, deep vein thrombosis and other complications occurred. Conclusion:Hybrid fixation with tibial prosthesis in primary TKA for patients with osteoporosis or osteopenia has a satisfactory clinical effect, which can relieve knee pain and improve joint function.

Objective To explore the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponin,and the UGT gene in Dipsacus asper Wall.ex Henry has been analyzed by the identification of whole genome,genome and prokaryotic expression.Methods The laboratory self-tested sequenced protein sequence files of the Dipsacus asper Wall.ex Henry genome were used.To validate the conserved domains of the sequence of the Dipsacus asper Wall.ex Henry UGT gene,BLASTP and hmmsearch were utilized.Prot-Param,SOMPA,MAGA7.0,Tbtools and other tools were used to investigate the protein physicochemical properties,protein structure,and covariance analysis of the Dipsacus asper Wall.ex Henry UGT gene family,and using the joint analysis of transcriptomic data and metabolomics data,two glycosyltransferases that might be related to triterpene saponin biosynthesis were screened,and expression vectors were constructed for prokaryotic expression.Results 44 Dipsacus asper Wall.ex Henry UGT genes were identified from the Dipsacus asper Wall.ex Henry genome.The length of Dipsacus asper Wall.ex Henry UGT proteins ranged from 49 to 1083 amino acids,with an average molecular weight of 24.86 kDa and an isoelectric point of 4.31-8.59.Dipsacus asper Wall.ex Henry UGT gene family was distributed on eight chromosomes.The phylogenetic tree constructed from the sequences of Dipsacus asper Wall.ex Henry,Arabidopsis thaliana and identified UGTs showed that glycosyltransferase gene families in Dipsacus asper Wall.ex Henry were mainly in the UGT73,UGT81,UGT85,and UGT80 families.Cis-acting element analysis showed that light-responsive elements were the most prevalent elements in the promoter regions of UGT gene family members.Two glycosyltransferases potentially related to triterpene saponin biosynthesis were screened using combined transcriptomics and metabolomics analysis,and were successfully expressed in prokaryotic form.Conclusion In this study,two candidate genes related to the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponins were jointly screened for prokaryotic expression using multi-omics,and were subjected to prokaryotic expression for further validation of the function of the genes.

Objective:To study the characteristics of soil microbial composition in sheep sheds in agricultural and pastoral areas, and explore potential zoonotic pathogen species.Methods:Using cross-sectional survey method, soil samples from sheep sheds in the agricultural and pastoral areas of Darhan-Muminggan Joint County, Baotou City, Inner Mongolia Autonomous Region were collected from August to October 2023. Genomic DNA was extracted, and metagenomic second-generation sequencing was conducted. Microbial species annotation was carried out by Kaiju method, and the soil microbial composition and zoonotic pathogen species of sheep sheds in agricultural and pastoral areas were analyzed.Results:A total of 10 and 5 soil samples were collected from sheep sheds in agricultural and pastoral areas, respectively. The results of β diversity analysis showed that there was a significant separation trend in soil samples from sheep sheds in agricultural and pastoral areas, and the interpretation of principal co-ordinates analysis (PCoA) 1 and PCoA2 were 27.8% and 17.4%, respectively. There was no significant difference in the microbial composition of soil samples from sheep sheds in agricultural and pastoral areas ( R = 0.09, P = 0.242). At the phylum level, the dominant bacterial phyla were Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. At the genus level, the dominant bacterial genera were Corynebacterium, Luteimonas, Atopostipes, and Salinicoccus. A variety of zoonotic pathogens were detected, including Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani in 15 soil samples. Conclusion:The soil microorganisms in sheep sheds in agricultural and pastoral areas are diverse, and zoonotic pathogens such as Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani are detected.