BACKGROUND:Osteoporotic fracture is the most serious complication of osteoporosis.Previous studies have demonstrated that gut microbiota has a regulatory effect on skeletal tissue and that gut microbiota has an important relationship with osteoporotic fracture,but the causal relationship between the two is unclear. OBJECTIVE:To explore the causal relationship between gut microbiota and osteoporotic fractures using Mendelian randomization method. METHODS:The genome-wide association study(GWAS)datasets of gut microbiota and osteoporotic fracture were obtained from the IEU Open GWAS database and the Finnish database R9,respectively.Using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,Mendelian randomization analyses with random-effects inverse variance weighted,MR-Egger regression,weighted median,simple model,and weighted model methods were performed to assess whether there is a causal relationship between gut microbiota and osteoporotic fracture.Sensitivity analyses were performed to test the reliability and robustness of the results.Reverse Mendelian randomization analyses were performed to further validate the causal relationship identified in the forward Mendelian randomization analyses. RESULTS AND CONCLUSION:The results of this Mendelian randomization analysis indicated a causal relationship between gut microbiota and osteoporotic fracture.Elevated abundance of Actinomycetales[odds ratio(OR)=1.562,95%confidence interval(CI):1.027-2.375,P=0.037),Actinomycetaceae(OR=1.561,95%CI:1.027-2.374,P=0.037),Actinomyces(OR=1.544,95%CI:1.130-2.110,P=0.006),Butyricicoccus(OR=1.781,95%CI:1.194-2.657,P=0.005),Coprococcus 2(OR=1.550,95%CI:1.068-2.251,P=0.021),Family ⅩⅢ UCG-001(OR=1.473,95%CI:1.001-2.168,P=0.049),Methanobrevibacter(OR=1.274,95%CI:1.001-1.621,P=0.049),and Roseburia(OR=1.429,95%CI:1.015-2.013,P=0.041)would increase the risk of osteoporotic fractures in patients.Elevated abundance of Bacteroidia(OR=0.660,95%CI:0.455-0.959,P=0.029),Bacteroidales(OR=0.660,95%CI:0.455-0.959,P=0.029),Christensenellacea(OR=0.725,95%CI:0.529-0.995,P=0.047),Ruminococcaceae(OR=0.643,95%CI:0.443-0.933,P=0.020),Enterorhabdus(OR=0.558,95%CI:0.395-0.788,P=0.001),Eubacterium rectale group(OR=0.631,95%CI:0.435-0.916,P=0.016),Lachnospiraceae UCG008(OR=0.738,95%CI:0.546-0.998,P=0.048),and Ruminiclostridium 9(OR=0.492,95%CI:0.324-0.746,P=0.001)would reduce the risk of osteoporotic fractures in patients.We identified 16 gut microbiota associated with osteoporotic fracture by the Mendelian randomization method.That is,using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,eight gut microbiota showed positive causal associations with osteoporotic fracture and another eight gut microbiota showed negative causal associations with osteoporotic fracture.The results of this study not only identify new biomarkers for the early prediction of osteoporotic fracture and potential therapeutic targets in clinical practice,but also provide an experimental basis and theoretical basis for the study of improving the occurrence and prognosis of osteoporotic fracture through gut microbiota in bone tissue engineering.

For middle-aged and elderly patients with conditions such as spinal fractures and degenerative spinal diseases, spinal internal fixation is a core surgical procedure for reconstructing spinal stability, heavily relying on the biomechanical stability provided by pedicle screw systems. Whereas, these patients are often complicated by osteoporosis that can significantly compromise the stability of the bone-pedicle screw interface, leading to a marked increase in pedicle screw loosening and surgical failure rates. The bone cement-augmented pedicle screw technique, which involves injecting bone cement into the vertebral body or screw trajectory to optimize the mechanical properties of the bone-pedicle screw composite, has been proven to significantly enhance fixation strength and effectively prevent screw-related failures, thereby reducing the incidence of internal fixation failure in high-risk populations undergoing spinal fusion. However, the widespread clinical application of this technique has faced challenges such as inaccurate clinical decision-making (indication and contraindication selection), non-standardized operative practices, and insufficient awareness of complication prevention, resulting in considerable variability in clinical outcomes and even severe complications. To address this, Prof. Luo Fei from First Affiliated Hospital of Army Medical University initiated the project and the Chinese Association Orthopaedic Surgeons organized relevant experts to develop the Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique ( version 2025), based on current evidence. The guidelines put forward 8 recommendations regarding the clinical value, scope of application, and operational standards of the technique, aiming to provide evidence-based medical support and technical standardization for clinical decision-making.

Objective:To establish a combined model of tumor heterogeneity metabolic parameters using 18F-FDG PET/CT and explore its value in differentiating benign from malignant pulmonary lesions. Methods:A total of 251 patients (157 males, 94 females; age 15-88 years) who were diagnosed with malignant lung lesions by 18F-FDG PET/CT and with definitive pathological results at Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from February 2017 to February 2024 were retrospectively enrolled. Analysis was conducted on clinical data, traditional parameters (SUV max, metabolic tumor volume (MTV), total lesion glycolysis (TLG)) of primary lesions on 18F-FDG PET/CT, and intra-tumoral metabolic heterogeneity index (HI; such as cumulative SUV volume histogram AUC (AUC-CSH), linear regression slope, CV). AUC-CSH and CV were calculated using SUV thresholds of 2.5 and 40%SUV max. Logistic univariate and multivariate regression analyses were used to extract independent predictors in clinical features and PET/CT parameters for the differential diagnosis of pulmonary lesions. A multi-parameter combined model was established through logistic regression and validated for diagnostic efficacy using ROC curve analysis. Results:Among 251 patients, 101 were benign and 150 were malignant. In univariate analysis, gender, age, tumor markers, spiculation sign, lobulation sign, vessel convergence sign, air bronchogram, long diameter, short diameter, SUV max, AUC-CSH 2.5, AUC-CSH 40%, CV2.5, and CV40% were predictive factors for the diagnosis of benign and malignant tumors (odds ratio ( OR): 0.57-17.39, all P<0.05). In multivariate analysis, gender, age, tumor markers, lobulation sign, vessel convergence sign, SUV max, AUC-CSH 40%, and CV40% were independent predictors for the diagnosis of benign and malignant tumors ( OR: 2.30-13.18, all P<0.05). The AUC, sensitivity, specificity, and accuracy of the multi-parameter combined model established with the above independent predictors were 0.89, 77.33%(116/150), 84.16%(85/101), 80.08%(201/251), respectively. Conclusion:18F-FDG PET/CT multi-parameter combined model has high value in the differentiation of benign and malignant pulmonary lesions.

Objective:To evaluate the potential value of 18×10 3 translocator protein (TSPO) radioligand ( N, N-diethy1-2-(2-(4-(2- 18F-fluoroethoxy) phenyl)-5, 7-dimethylpyrazolo[1, 5-A]pyrimidin-3-yl)acetamide, 18F-DPA-714) PET compared with conventional MR in the detection of autoimmune encephalitis (AE), the correlation with clinical symptoms, and the monitoring of immunotherapy efficacy in patients with AE. Methods:From December 2021 to June 2024, 45 AE patients (17 males, 28 females, age (38.3±17.0) years) diagnosed at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine and 10 healthy volunteers (7 males, 3 females, age (28.7±5.1) years) were enrolled in this prospective study. All participants underwent baseline 18F-DPA-714 PET/MR scans, and 23 of these AE patients underwent further follow-up 18F-DPA-714 PET/MR scans. 18F-DPA-714 PET positivity was defined as having an uptake intensity threshold higher than the mean SUV ratio (SUVR)+ 2 s of the corresponding brain region in healthy controls. MR positivity was defined as abnormal hyperintensity in a specific brain region or multiple brain regions on the T 2 fluid attenuated inversion recovery (FLAIR). The positive detection rates of 18F-DPA-714 PET and MR was analyzed using McNemar χ2 test, and the differences in the uptake intensity (SUVR) of 18F-DPA-714 between symptomatic and non-symptomatic groups, and between remission and non-remission groups after immunotherapy were compared using independent-sample t test or Wilcoxon rank sum test. Spearman rank correlation analysis was used to analyze the correlation between the changing rate of SUVR and the changing of the modified Rankin Scale (mRS) score before and after treatment. Results:The positive detecting rate of 18F-DPA-714 PET for AE was significantly higher than that of MR (73.3%(33/45) vs 35.6%(16/45); χ2=11.56, P=0.001). The cerebellar SUVR of ataxia patients was significantly higher than that of asymptomatic patients (1.22(1.06, 1.33) vs 1.08(0.99, 1.20); Z=-2.14, P=0.034). Follow-up imaging showed that the SUVR of patients in the remission group after immunotherapy was significantly lower than that in the non-remission group ((-15.19±10.17)% vs (14.26±13.36)%; t=5.81, P<0.001). There was a significant correlation between the changing rate of SUVR and the changing of the mRS score before and after treatment ( rs=0.65, P<0.001). Conclusion:Compared with conventional MR, 18F-DPA-714 PET has a higher positive detecting rate for AE, and has the potential to reflect the clinical symptoms of AE and monitor the efficacy of immunotherapy.

Objective:To investigate the correlation between thyroid function and the total cerebral small vessel disease score in the elderly.Methods:This cross-sectional study included elderly people who underwent physical examinations at the Geriatrics Department of Beijing Hospital from April 2019 to December 2020. Participants were devided into 5 groups based on the total score of cerebral small vessel disease: 0, 1, 2, 3, and 4. General clinical data were collected through physical examination reports and outpatient medical records. All participants underwent a 3T brain magnetic resonance imaging (MRI), scan and data were collected to calculate the total cerebral small vessel disease score. Fasting venous blood samples were taken in the morning to measure thyroid hormone levels. Relationships between thyroid hormone levels and lacunar infarcts, microbleeds, white matter hyperintensities (WMH), enlarged perivascular spaces, and total cerebral small vessel disease score were analyzed using Spearman correlation. Multivariate ordinal logistic regression was also used to analyze factors associated with total cerebral small vessel disease score.Results:A total of 545 elderly individuals were included, with a mean age of (75.16±9.18) years, and 505 (92.7%) were male. The number of subjects with a total cerebral small vessel disease score of 0 were 207 (38.0%), 1 were 182 (33.4%), 2 were 99 (18.2%), 3 were 41 (7.5%), and 4 were 16 (2.9%). Statistical significant differences were found in age, fasting blood glucose, systolic blood pressure, triiodothyronine (TT3), free triiodothyronine (FT3), and the proportion of hypertensive patients among the groups (all P<0.05). Spearman correlation analysis showed that TT3 ( r=-0.138, P=0.001) and FT3 ( r=-0.213, P<0.001) were negatively correlated with total cerebral small vessel disease score. Multivariate ordinal logistic regression analysis revealed that age was independently and positively associated with total cerebral small vessel disease score ( OR=1.139, 95% CI: 1.087-1.193, P<0.001), while FT3 was independently and negatively associated with total cerebral small vessel disease score ( OR=0.331, 95% CI: 0.118-0.929, P=0.009). Conclusion:In the elderly population, FT3 levels are independently and negatively associated with total cerebral small vessel disease score.

Objective:To assess the association of single nucleotide polymorphisms of rs700519 and rs4646 loci of cytochrome P450 19A1 ( CYP19A1) gene with risk of breast cancer. Methods:Two hundred patients with breast cancer treated at Xinxiang Central Hospital between January 2019 and January 2024 and 100 healthy individuals were enrolled as the study group and control group, respectively. The genotypes of the CYP19A1 gene at the rs700519 and rs4646 loci were determined by direct sequencing. The general data, distribution of CYP19A1 genotypes and alleles were compared between the two groups. This study has been approved by the Medical Ethics Committee of Xinxiang Central Hospital (Ethics No.2021-182). Results:No significant difference was found in age, body mass index, times of conception and proportion of menopause between the two groups ( P>0.05). The frequencies of AA genotype and A allele at the rs700519 locus, and the CC genotype and C allele at the rs4646 locus in the study group were significantly higher than those of the control group ( P<0.05). The frequencies of AA genotype at the rs700519 locus and CC genotype at the rs4646 locus in patients with breast cancer at stages Ⅲ-Ⅳ were significantly higher than those at stage Ⅰ-Ⅱ ( P<0.05). Conclusion:Polymorphisms of CYP19A1 gene at the rs700519 and rs4646 loci are associated with susceptibility of breast cancer. The AA and CC genotypes at the two loci may increase the risk for breast cancer.

Objective:To establish a rapid and accurate duplex real-time fluorescent reverse transcription-recombinase polymerase amplification/recombinase-aided amplification（RT-RPA/RAA）detection method for identification and differentiation of Nipah virus（NiV），Langya virus（LayV），Mojiang virus（MoJV），and Cedar virus（CedV）.Methods:First，specific primers and probes were designed targeting the conserved L gene regions of NiV and LayV，as well as the conserved N gene regions of MoJV and CedV，respectively. The four viruses were divided into two groups for duplex detection. Subsequently，the optimal primer and probe combinations were screened by comparing the amplification efficiency of different primer pair combinations（F1/R1，F1/R2，F2/R1，F2/R2）. The reaction temperature was optimized through temperature gradient settings from 37 ℃ to 42 ℃，and the amounts of primers and probes were optimized to establish the duplex real-time fluorescent RT-RPA/RAA detection system. Finally，the detection performance was evaluated through specificity，sensitivity，and stability tests，as well as clinical sample validation.Results:The selected primer pairs（NiV primer pair F2/R1，LayV primer pair F1/R1，MoJV primer pair F2/R2，and CedV primer pair F2/R2）all demonstrated optimal amplification efficiency when combined with their corresponding probes. The optimal annealing temperature was 39 ℃，and the minimum detection limit was 101-102 copies/μl. The method could effectively distinguish target viruses from other non-target viruses，and repeated experiments showed good stability（ R2> 0.90）. Additionally，detection results for Malaysian NiV strains and various clinical samples were consistent with the Taqman multiplex qRT-PCR method. Conclusion:The duplex real-time fluorescent RT-RPA/RAA detection method successfully established in this study can rapidly and accurately identify and differentiate four important henipavirus-like viruses：NiV，LayV，MoJV，and CedV. It features simple operation，rapid reaction，high specificity，and good stability，providing an effective molecular detection tool for rapid field diagnosis，surveillance，and control of these zoonotic viruses.

Objective:To evaluate the prognostic value of 18F-fluorodeoxyglucose ( 18F-FDG) PET/CT metabolic parameters in small cell lung cancer (SCLC) . Methods:A retrospective analysis was conducted on the clinical and imaging data of 156 SCLC patients, who underwent 18F-FDG PET/CT imaging and were diagnosed by histopathological examination at Nanjing Drum Tower Hospital, Affiliated Hospital of Nanjing University Medical School from September 2013 to February 2024. The metabolic tumor volume (MTV), total lesion glycolysis (TLG), linear regression slope, area under the curve of cumulative standard uptake value (SUV) volume histogram (AUC-CSH), and coefficient of variation (CV) were calculated using LIFEx software with different SUV thresholds. Univariate and multivariate analyses were performed using Cox proportional hazards model. Patient stratification was based on the critical values determined by receiver operator characteristic (ROC) curve analysis. The survival curve was plotted using the Kaplan-Meier method and log-rank test was performed. Results:Univariate analysis showed that MTV 40% ( HR=2.91, 95% CI: 1.55-5.47, P=0.001), MTV 60% ( HR=2.31, 95% CI: 1.29-4.17, P=0.005), TLG 40% ( HR=2.07, 95% CI: 1.19-3.60, P=0.010), linear regression slope ( HR=0.45, 95% CI: 0.26-0.79, P=0.005), and CV 40% ( HR=0.27, 95% CI: 0.08-0.84, P=0.024) were factors affecting progression-free survival (PFS) in SCLC patients. MTV 40% ( HR=1.98, 95% CI: 1.22-3.22, P=0.005), MTV 60% ( HR=1.80, 95% CI: 1.12-2.88, P=0.015), MTV 80% ( HR=1.71, 95% CI: 1.08-2.74, P=0.024), TLG 40% ( HR=3.68, 95% CI: 1.59-8.49, P=0.002), linear regression slope ( HR=0.49, 95% CI: 0.30-0.80, P=0.004), and AUC-CSH 80% ( HR=0.44, 95% CI: 0.23-0.84, P=0.013) were found to be factors affecting overall survival (OS) in SCLC patients. Multivariate analysis revealed that MTV 40% ( HR=4.76, 95% CI: 1.11-20.50, P=0.036) was an independent factor influencing PFS, and TLG 40% ( HR=3.19, 95% CI: 1.02-9.92, P=0.046) was an independent factor influencing OS in SCLC patients. ROC curve analysis identified the optimal cutoff value for MTV 40% in predicting PFS as 5.5cm 3 and the optimal cutoff value for TLG 40% in predicting OS as 41.5 g in SCLC patients. Survival analysis showed that patients with MTV 40%≤5.5 cm 3 ( n=33) had a median PFS that was not reached, while patients with MTV 40%>5.5 cm 3 ( n=123) had a median PFS of 10.3 months, with a statistically significant difference ( χ2=12.09, P=0.001). For patients with TLG 40%≤41.5 g ( n=35), the median OS was not reached, whereas for TLG 40%>41.5 g ( n=121), the median OS was 31.6 months, with a statistically significant difference ( χ2=10.55, P=0.001) . Conclusions:The 18F-FDG PET/CT metabolic parameter MTV 40% is an independent factor influencing PFS, while TLG 40% is an independent factor influencing OS in SCLC patients. The above two parameters may serve as indicators for assessing the prognosis of SCLC patients.

Objective:To explore the deubiquitination modification of the deubiquitinase BRCC3 in rats with neuropathic pain (NPP) treated with meridian tuina of Zhuang medicine and its analgesic molecular mechanism.Methods:Rats were divided into normal group, sham-operation group, model group, sham-tuina group, and meridian tuina of Zhuang medicine group using a random number table method, with 9 rats in each group. Except for the normal and sham operation groups, spinal nerve ligation models were prepared in all other groups. On the first day after surgery, intervention was carried out on the meridian tuina of Zhuang medicine and sham-tuina groups for 14 days, while the other three groups were not intervened; the paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) of each group of rats were measured at 0 days before operation, 1, 7, and 14 days after operation. Western blot was used to detect the expressions of BRCC3 and NLRP3 proteins in spinal cord tissue, and ELISA was used to detect the level of IL-1β in serum.Results:On postoperative 7 and 14 d, compared with the model group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLPR3 and BRCC3 expression in spinal cord tissue, and a decrease in serum IL-1β levels ( P<0.05). Compared with the sham-tuina group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLRP3 protein expression in spinal cord tissue, and a decrease in IL-1β levels in serum ( P<0.05). Conclusion:Meridian tuina of Zhuang medicine can alleviate pain sensitivity in SNL model rats, and its mechanism is related to the inhibition of the expression of deubiquitinase BRCC3 by meridian massage of Zhuang medicine, which increases the ubiquitination level of NLRP3 and hinders its activation, thereby blocking the immune inflammatory response mediated by inflammatory factors.

Objective To investigate the protective effects of paeonol(PAE)on autophagy in human neuroblastoma cells(SH-SY5Y)induced by overexpression of α-synuclein(α-Syn),and to explore its related mechanism.Methods SH-SY5Y cells served as control group,while those induced with A53T-α-Syn mutation were used as model group.Additional groups included PAE(150 μg/ml)group,3-MA(1 mmol/L)group,and PAE(150 μg/ml)+3-MA(1 mmol/L)group.Cell viability was assessed using CCK-8 method,cell morphology was observed under an optical microscope,and protein expressions of α-Syn,LC3-Ⅱ,p62,Beclin-1,phosphorylated c-Jun N-terminal kinase(p-JNK),and p-Bcl-2 were determined by Western blotting.Results Compared with control group,model control exhibited decreased cell survival(P＜0.01),increased α-Syn expression(P＜0.001),reduced expression of autophagy-related proteins LC3-Ⅱ and Beclin-1(P＜0.01,P＜0.05),elevated autophagy substrate protein p62(P＜0.05),and decreased expression of autophagy pathway-related proteins p-JNK and Bcl-2(P＜0.05,P＜0.01).Compared with model group,PAE group showed increased cell survival(P＜0.01),decreased α-Syn and p62 protein expression(P＜0.01,P＜0.05),and increased expression of LC3-Ⅱ,Beclin-1,p-JNK and Bcl-2(P＜0.05).Compared with PAE group,3-MA+PAE group demonstrated increased α-Syn expression(P＜0.05).Conclusions PAE could attenuate the injury of SH-SY5Y cells induced by A53T-α-Syn and eliminate over-expressed α-Syn by activating autophagy pathway,which may be associated with the upregulation of JNK/Bcl-2 mediated autophagy pathway.