Objective:To investigate the diagnostic value of metagenomic next-generation sequencing (mNGS) for pyogenic spinal infections.Methods:A total of 255 patients diagnosed with pyogenic spinal infections were enrolled between September 2022 and September 2024 at Qingdao University Affiliated Hospital, Fuzhou Second General Hospital, First Affiliated Hospital of Nanchang University, Shandong University Affiliated Public Health Clinical Center, and the Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Among them, 155 were male and 100 were female, with an average age of 62.5±14.2 years (ranging from 13 to 90 years). All patients had samples of infected tissue and/or pus collected for microbial culture and mNGS testing. The number, types, and positive rates of pathogens detected by microbial culture and mNGS were compared. Using culture results as the gold standard, receiver operating characteristic (ROC) curve analysis was performed for mNGS testing and the combined method of mNGS and microbial culture, calculating the area under the curve (AUC) and 95% CI. Results:All 255 cases were clinically diagnosed as pyogenic spinal infections, with 194 cases providing microbiological evidence. The most common Gram-positive bacterium was Staphylococcus aureus, while the most common Gram-negative bacterium was Escherichia coli. A total of 33 pathogenic microorganisms were detected by mNGS, while microbial culture detected 18 pathogenic microorganisms. The positive rate of mNGS was 72.2% (184 out of 255), which was significantly higher than that of 30.2% (77 out of 255) for microbial culture, showing a significant difference (χ 2=90.150, P<0.001); the positive rate of mNGS combined with microbial culture was 76.1% (194 out of 255) with significant difference compared to mNGS alone (χ 2=8.100, P<0.001). Among 178 culture-negative samples, the detection rate of mNGS was 65.7% (117 out of 178); among 77 culture-positive samples, the detection rate of mNGS was 87.0% (67 out of 77), and 97.0% (65 out of 67) of the detected pathogens matched the culture results at the species level. The AUCs of the ROC curves for mNGS testing and the combination of mNGS with microbial culture were 0.606 [95% CI (0.534, 0.678)] and 0.671 [95% CI (0.606, 0.736)], respectively, with significant differences compared to microbial culture ( P=0.007; P=0.007). Conclusions:mNGS demonstrates superior performance over conventional culture in identifying pathogens in pyogenic spinal infections. Moreover, combining mNGS with culture further improves diagnostic yield, supporting its integration into clinical practice.

Clinical data of two patients with X-linked adrenoleukodystrophy (X-ALD) initially presenting as Addison′s disease were collected from the Department of Endocrinology, First Medical Center of Chinese PLA General Hospital. Relevant medical history, clinical features, laboratory tests, and genetic results were analyzed. The two male patients, aged 7 years (case 1) and 15 years (case 2), initially presented with generalized skin hyperpigmentation, without any family history of similar disorders. Both had normal growth and development, and adrenal CT and brain MRI revealed no significant abnormalities. Elevated very long-chain fatty acid (VLCFA) levels were detected. Genetic analyses identified a maternally inherited missense mutation (c.830G>A, p.Gly277Glu) in the ATP-binding cassette subfamily D member 1 (ABCD1) gene in case 1, and a missense mutation (c.1499G>T, p.Gly500Val) in case 2. Protein structural predictions indicated both mutations as potentially damaging or damaging, and both were classified as likely pathogenic according to American College of Medical Genetics and Genomics (ACMG) criteria (PM1/PM2/PP3_Moderate and PM2/PP3_Moderate/PM6, respectively), supporting their correlation with the clinical phenotype. Clinicians should maintain vigilance for X-ALD in male patients presenting with Addison′s disease, and combined VLCFA and genetic testing can effectively prevent misdiagnosis or delayed diagnosis.

Objective To compare the application effects of C-shaped forceps external fixation emergency technique and pelvic belt suspension traction technique in unstable pelvic posterior ring fractures.Methods A total of 80 patients with unstable pelvic posterior ring fractures were selected and randomly divided into the observation group and the control group,with 40 cases in each group.The control group was given pelvic belt suspension traction technique to fix pelvic fractures,while the observation group was given C-shaped forceps external fixation emergency technique to fix pelvic fractures.After fixation,pelvic X-ray examination was performed to evaluate the reduction quality of pelvic fractures in two groups.The emergency operation/surgery time,total blood transfusion volume and 2-hour shock index after surgery were recorded.The emergency effects(rebleeding rate,secondary injury rate,secondary surgery rate,rescue success rate and internal fixation surgery rate)were evaluated.The facial expression pain scale(FPS-R)was used to assess the degree of pain before fixation and 30 minutes after fixation.After 6 months of treatment,pelvic function was evaluated using the Pelvic Fracture Function Rating System(Majeed).Results The excellent rate of pelvic fracture reduction quality was higher in the observation group than that in the control group(75.0%vs.55.0%,P＜0.05).The secondary injury rate,internal fixation surgery rate and secondary surgery rate were lower in the observation group than those of the control group,and the success rate of rescue was higher than that of the control group(P＜0.05).The emergency operation/surgery time,total blood transfusion volume and shock index were lower in the observation group than those in the control group(P＜0.05).After 30 minutes of fixation,the FPS-R scores of both groups decreased compared to before fixation,and FPS-R scores of the observation group was lower than those of the control group(P＜0.05).After 6 months of treatment,Majeed scores in both groups were increased compared to those before fixation,and there were no significant differences in Majeed scores between the two groups.Conclusion The emergency treatment of unstable pelvic posterior ring fractures using C-shaped forceps external fixation technique has shown good results and is worthy of clinical promotion and application.

Objective To investigate the impact of chronic observational social defeat stress on multidimensional behaviors in female mice,including social interaction,emotion,and cognition,as well as the underlying molecular mechanisms.Methods Adult female C57BL6N mice were divided into a stress group and a control group.Behavioral assessment were conducted through social interaction tests,anxiety-like behavior tests,and cognitive tests(novel object recognition and spatial memory).Synaptic plasticity-related proteins in the hippocampus were detected using Western blot.Results Behavioral tests revealed that the stress group showed reduced social interaction time compared to the control group[(106.2±9.31)s vs.(156.5±14.81)s,P<0.05].In a novel environment,the stress group consumed less food than the control group[(0.10±0.01)g vs.(0.16±0.02)g,P<0.05].Cognitive tests demonstrated that the stress group exhibited impaired novel object recognition[(52.17%%±2.67%)vs.(65.91%±5.23%)]and spatial memory[(61.84%±3.14%)vs.(74.13%±1.94%)]compared to the control group(P<0.05).Western blot analysis showed that the expression of hippocampal synaptic plasticity-related proteins,including the glutamate ionotropic receptor N-methyl-D-aspartate(NMDA)receptor subunit NR2A[(0.66±0.05)vs.(0.89±0.08)]and brain-derived neurotrophic factor(BDNF)[(0.81±0.05)vs.(1.14±0.06)],was significantly decreased in the stress group(P<0.05).Conclusion Chronic observational social defeat stress induces social withdrawal,anxiety-like behaviors,and cognitive deficits in female mice.These effects may be associated with reduced expression of NR2A and BDNF in the hippocampus.

Objective:To investigate the diagnostic value of metagenomic next-generation sequencing (mNGS) for pyogenic spinal infections.Methods:A total of 255 patients diagnosed with pyogenic spinal infections were enrolled between September 2022 and September 2024 at Qingdao University Affiliated Hospital, Fuzhou Second General Hospital, First Affiliated Hospital of Nanchang University, Shandong University Affiliated Public Health Clinical Center, and the Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Among them, 155 were male and 100 were female, with an average age of 62.5±14.2 years (ranging from 13 to 90 years). All patients had samples of infected tissue and/or pus collected for microbial culture and mNGS testing. The number, types, and positive rates of pathogens detected by microbial culture and mNGS were compared. Using culture results as the gold standard, receiver operating characteristic (ROC) curve analysis was performed for mNGS testing and the combined method of mNGS and microbial culture, calculating the area under the curve (AUC) and 95% CI. Results:All 255 cases were clinically diagnosed as pyogenic spinal infections, with 194 cases providing microbiological evidence. The most common Gram-positive bacterium was Staphylococcus aureus, while the most common Gram-negative bacterium was Escherichia coli. A total of 33 pathogenic microorganisms were detected by mNGS, while microbial culture detected 18 pathogenic microorganisms. The positive rate of mNGS was 72.2% (184 out of 255), which was significantly higher than that of 30.2% (77 out of 255) for microbial culture, showing a significant difference (χ 2=90.150, P<0.001); the positive rate of mNGS combined with microbial culture was 76.1% (194 out of 255) with significant difference compared to mNGS alone (χ 2=8.100, P<0.001). Among 178 culture-negative samples, the detection rate of mNGS was 65.7% (117 out of 178); among 77 culture-positive samples, the detection rate of mNGS was 87.0% (67 out of 77), and 97.0% (65 out of 67) of the detected pathogens matched the culture results at the species level. The AUCs of the ROC curves for mNGS testing and the combination of mNGS with microbial culture were 0.606 [95% CI (0.534, 0.678)] and 0.671 [95% CI (0.606, 0.736)], respectively, with significant differences compared to microbial culture ( P=0.007; P=0.007). Conclusions:mNGS demonstrates superior performance over conventional culture in identifying pathogens in pyogenic spinal infections. Moreover, combining mNGS with culture further improves diagnostic yield, supporting its integration into clinical practice.

Objective To compare the application effects of C-shaped forceps external fixation emergency technique and pelvic belt suspension traction technique in unstable pelvic posterior ring fractures.Methods A total of 80 patients with unstable pelvic posterior ring fractures were selected and randomly divided into the observation group and the control group,with 40 cases in each group.The control group was given pelvic belt suspension traction technique to fix pelvic fractures,while the observation group was given C-shaped forceps external fixation emergency technique to fix pelvic fractures.After fixation,pelvic X-ray examination was performed to evaluate the reduction quality of pelvic fractures in two groups.The emergency operation/surgery time,total blood transfusion volume and 2-hour shock index after surgery were recorded.The emergency effects(rebleeding rate,secondary injury rate,secondary surgery rate,rescue success rate and internal fixation surgery rate)were evaluated.The facial expression pain scale(FPS-R)was used to assess the degree of pain before fixation and 30 minutes after fixation.After 6 months of treatment,pelvic function was evaluated using the Pelvic Fracture Function Rating System(Majeed).Results The excellent rate of pelvic fracture reduction quality was higher in the observation group than that in the control group(75.0%vs.55.0%,P＜0.05).The secondary injury rate,internal fixation surgery rate and secondary surgery rate were lower in the observation group than those of the control group,and the success rate of rescue was higher than that of the control group(P＜0.05).The emergency operation/surgery time,total blood transfusion volume and shock index were lower in the observation group than those in the control group(P＜0.05).After 30 minutes of fixation,the FPS-R scores of both groups decreased compared to before fixation,and FPS-R scores of the observation group was lower than those of the control group(P＜0.05).After 6 months of treatment,Majeed scores in both groups were increased compared to those before fixation,and there were no significant differences in Majeed scores between the two groups.Conclusion The emergency treatment of unstable pelvic posterior ring fractures using C-shaped forceps external fixation technique has shown good results and is worthy of clinical promotion and application.

Objective To investigate the impact of chronic observational social defeat stress on multidimensional behaviors in female mice,including social interaction,emotion,and cognition,as well as the underlying molecular mechanisms.Methods Adult female C57BL6N mice were divided into a stress group and a control group.Behavioral assessment were conducted through social interaction tests,anxiety-like behavior tests,and cognitive tests(novel object recognition and spatial memory).Synaptic plasticity-related proteins in the hippocampus were detected using Western blot.Results Behavioral tests revealed that the stress group showed reduced social interaction time compared to the control group[(106.2±9.31)s vs.(156.5±14.81)s,P<0.05].In a novel environment,the stress group consumed less food than the control group[(0.10±0.01)g vs.(0.16±0.02)g,P<0.05].Cognitive tests demonstrated that the stress group exhibited impaired novel object recognition[(52.17%%±2.67%)vs.(65.91%±5.23%)]and spatial memory[(61.84%±3.14%)vs.(74.13%±1.94%)]compared to the control group(P<0.05).Western blot analysis showed that the expression of hippocampal synaptic plasticity-related proteins,including the glutamate ionotropic receptor N-methyl-D-aspartate(NMDA)receptor subunit NR2A[(0.66±0.05)vs.(0.89±0.08)]and brain-derived neurotrophic factor(BDNF)[(0.81±0.05)vs.(1.14±0.06)],was significantly decreased in the stress group(P<0.05).Conclusion Chronic observational social defeat stress induces social withdrawal,anxiety-like behaviors,and cognitive deficits in female mice.These effects may be associated with reduced expression of NR2A and BDNF in the hippocampus.

Clinical data of two patients with X-linked adrenoleukodystrophy (X-ALD) initially presenting as Addison′s disease were collected from the Department of Endocrinology, First Medical Center of Chinese PLA General Hospital. Relevant medical history, clinical features, laboratory tests, and genetic results were analyzed. The two male patients, aged 7 years (case 1) and 15 years (case 2), initially presented with generalized skin hyperpigmentation, without any family history of similar disorders. Both had normal growth and development, and adrenal CT and brain MRI revealed no significant abnormalities. Elevated very long-chain fatty acid (VLCFA) levels were detected. Genetic analyses identified a maternally inherited missense mutation (c.830G>A, p.Gly277Glu) in the ATP-binding cassette subfamily D member 1 (ABCD1) gene in case 1, and a missense mutation (c.1499G>T, p.Gly500Val) in case 2. Protein structural predictions indicated both mutations as potentially damaging or damaging, and both were classified as likely pathogenic according to American College of Medical Genetics and Genomics (ACMG) criteria (PM1/PM2/PP3_Moderate and PM2/PP3_Moderate/PM6, respectively), supporting their correlation with the clinical phenotype. Clinicians should maintain vigilance for X-ALD in male patients presenting with Addison′s disease, and combined VLCFA and genetic testing can effectively prevent misdiagnosis or delayed diagnosis.

Objective:To explore the role of metagenomic next-generation sequencing (mNGS) in the diagnosis of pathogens in primary infectious diseases of the spine (IDS) and to reveal its pathogen spectrum.Methods:This is a retrospective multi-center case series study. Clinical data of 380 patients with primary IDS who were treated at four medical centers in China from December 2019 to April 2024 were retrospectively analyzed. Among them, 82 cases were from the Department of Spine Surgery at the Affiliated Hospital of Qingdao University, 129 cases were from the Orthopedics Section Ⅱ (Bone Infection), Public Health Clinical Center Affiliated to Shandong University, 112 cases were from the Department of Spine Surgery, Fuzhou Second General Hospital, and 57 cases were from the Department of Orthopedic Surgery, Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. There were 238 males and 242 females, with an age of (61.4±13.1) years (range: 10 to 91 years). Specimens from the site of spinal infection were obtained for pathogen culture, pathological examination, and mNGS detection preoperatively or intraoperatively in all patients. The number, types, and positive rates of pathogens detected by the two methods were analyzed and compared using the Chi-square test.Results:Among the 380 patients, 320 had confirmed pathogenic bacteria, with the highest proportion being pyogenic bacterial infections, accounting for 76.9% (246/320). The most common pathogen was Staphylococcus aureus, accounting for 22.8% (73/320). Brucella accounted for 13.8% (44/320); Mycobacterium tuberculosis accounted for 6.3% (20/320). Fungal infections accounted for 3.4% (11/320), mainly Aspergillus and Candida. In addition, Mycoplasma was detected in 3 cases (0.9%) and Benacox body in 4 cases (1.2%). The pathogen spectrum constructed by mNGS covered 46 types of pathogens, higher than the 22 types detected by traditional methods. The positive rate of mNGS was 80.8% (308/381), significantly higher than the 27.9% (106/381) of traditional methods ( χ2=182.53, P<0.01). Conclusions:mNGS improves the positive rate of pathogen diagnosis in IDS, detecting a broader spectrum of pathogens, and serves as a valuable complement to traditional diagnostic methods. Combining both methods in the diagnosis of IDS can maximize detection rates, providing robust evidence for precise anti-infective treatment.

Objective: To investigate the differences in enolase and laminin levels in vaginal epithelial tissues between mice successfully infected withGardnerellaand mice not infected with Gardnerella, providing information for further exploration of the correlation between enolase and laminin levels and the incidence of bacterial vaginosis. Methods: Gardnerella strains isolated, purified, and identified from vaginal secretions of patients with bacterial vaginosis were used to infect the vagina of mice and establish a mouse model of bacterial vaginosis. Successful and failed mice was defined as successful and failed groups, respectively. Differential expression of enolase and laminin in the vaginal epithelial tissue of two groups of mice was detected by Western blot. Modeling success rate was statistically analyzed, and the expression differences of enolase and laminin was compared between two groups. Results: One strain of Gardnerella vaginalis infected 10 SPF grade KM mice, 7 mice met the diagnostic criteria for bacterial vaginosis, and 3 mice failed to model, with a success rate of 70%. Western blot was used to detect protein expression levels, and the levels of laminin and enolase in the successfully modeled mouse vaginal epithelial tissue were significantly higher than those in the failed modeling group, with statistical differences between the two groups(P<0.05). Conclusion Enolase and laminin may be involved in the occurrence of bacterial vaginosis, however, further research is needed to determine the mechanisms through which they trigger the occurrence and development of the disease.