Objective:To investigate the clinical significance and inducing factors of abnormal intraoperative neurophysiological monitoring (IONM) signals during surgery for cervical degenerative diseases.Methods:A retrospective analysis was performed on 586 patients who underwent cervical degenerative disease surgery with IONM at the Department of Orthopedics, The First Affiliated Hospital of Soochow University, from April 2015 to April 2024. Surgical approaches included 380 anterior spinal canal decompression and fusion procedures, 154 posterior spinal canal decompression and fusion procedures (including single-door laminoplasty, total laminectomy, and hemilaminectomy), and 52 combined anterior-posterior surgeries. The multimodal IONM protocol employed transcranial electrical stimulation motor evoked potentials (TES-MEP) and cortical somatosensory evoked potentials (CSEP), combined with electromyography (EMG). Bilateral deltoid muscles, thenar/hypothenar muscles and abductor hallucis muscles were monitored in all patients. Intraoperative MEP, SEP, and EMG results were recorded to analyze the causes of abnormal signals, intraoperative response strategies, and postoperative neurological function and outcomes. Fourfold table chi-square tests were used to analyze factors possibly associated with IONM alerts.Results:Among the 586 cervical surgeries, 17 cases (2.9%) exhibited abnormal IONM signals. These included 4 cases of anterior cervical discectomy and fusion (ACDF), 4 cases of anterior cervical corpectomy and fusion (ACCF), and 2 cases of combined anterior-posterior surgeries for cervical spondylotic myelopathy; and 5 posterior surgeries and 2 anterior ACCF procedures for ossification of the posterior longitudinal ligament (OPLL). The rate of abnormal IONM signals was significantly higher in patients with maximum spinal cord compression (MSCC)>60% (5.8%, 12/208) than in those with MSCC≤60% (χ 2=9.417, P=0.002); in patients with intraoperative hypotension during posterior surgery (mean arterial pressure reduction>20% from baseline, cumulative duration>20 min), the abnormal IONM rate was 22.2% (6/27), which was significantly higher than that in patients without intraoperative hypotension (χ 2=33.542, P<0.001); in patients who underwent calcified tissue removal during anterior surgery, the abnormal IONM rate was 9.3% (5/54), which was significantly higher than that in patients without calcified tissue removal (χ 2=13.162, P=0.003). Thus, MSCC>60%, intraoperative hypotension during posterior surgery, and calcified tissue removal during anterior surgery may be inducing factors for abnormal IONM signals. Among the 17 patients with monitoring abnormalities, 8 cases showed no significant improvement after corresponding intraoperative treatments, and 7 of these 8 cases experienced varying degrees of muscle strength decline and sensory numbness immediately after surgery; 9 cases showed partial or complete recovery of signals, among which 8 cases had no new-onset neurological impairment after surgery, and 1 case developed unilateral upper limb grip strength decline. IONM demonstrated a sensitivity of 0.8750 and specificity of 0.8889. Conclusions:Multimodal IONM can detect electrophysiological abnormalities of spinal cord nerve function during cervical degenerative disease surgery, providing real-time warning of potential nerve damage during the operation. The proportion of abnormal IONM signals is relatively high in cases with MSCC>60%, intraoperative hypotension during posterior cervical surgery, or calcified tissue removal during anterior cervical surgery.

BACKGROUND:Restoring the normal level of reactive oxygen species and mitochondrial function of nucleus pulposus cells and inhibiting apoptosis of nucleus pulposus cells are key targets for delaying intervertebral disc degeneration.Prussian blue nanoparticles have peroxidase-like activity,which can effectively remove reactive oxygen species in the pathological microenvironment and protect nucleus pulposus cells from oxidative stress damage.OBJECTIVE:To investigate the biological functions and mechanisms of Prussian blue nanoparticles in delaying nucleus pulposus degeneration in rats.METHODS:Prussian blue nanoparticles were prepared by hydrothermal method,and their micromorphology and particle size were characterized.Prussian blue nanoparticles with different mass concentrations(20,40,60,80,and 100 μg/mL)were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.Cell proliferation was detected by CCK-8 assay after 24 hours.60 μg/mL Prussian blue nanoparticles were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.The viability of nucleus pulposus cells was observed by live-dead staining after 1 and 3 days.Passage 2 SD rat caudal vertebrae nucleus pulposus cells were obtained and observed for cell adhesion before being divided into three intervention groups.The control group did not receive any intervention.The lipopolysaccharide group was added with lipopolysaccharide.The lipopolysaccharide+Prussian blue nanoparticle group was added with lipopolysaccharide and 60 μg/mL Prussian blue nanoparticles.Reactive oxygen species,mitochondrial superoxide,and mitochondrial membrane potential were detected 24 hours after intervention.RT-qPCR detection and Alcian blue staining were performed 48 hours after intervention.RESULTS AND CONCLUSION:(1)Under transmission electron microscopy,Prussian blue nanoparticles were uniform nanocubes with an average particle size of 130 nm.(2)CCK-8 assay results showed that 20-60 μg/mL Prussian blue nanoparticles had no obvious cytotoxicity,and 60 μg/mL Prussian blue nanoparticles were selected for cell intervention in subsequent experiments.Live-dead staining results showed that 60 μg/mL Prussian blue nanoparticles had no effect on the viability of nucleus pulposus cells.(3)Compared with the control group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide group were increased(P＜0.01),the mitochondrial membrane potential was decreased(P＜0.01),the mRNA expressions of type Ⅱ collagen and aggrecan were decreased(P＜0.01),the mRNA expressions of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 were increased(P＜0.01),and the positive area of Alcian blue staining was reduced(P＜0.01).Compared with the lipopolysaccharide group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide+Prussian blue nanoparticle group were decreased(P＜0.01),mitochondrial membrane potential increased(P＜0.01),mRNA expression of type Ⅱ collagen and aggrecan increased(P＜0.01),mRNA expression of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 decreased(P＜0.01),and positive area of Alcian blue staining increased(P＜0.01).The results showed that Prussian blue nanoparticles delayed the degeneration of rat nucleus pulposus by reducing oxidative stress of nucleus pulposus cells,restoring mitochondrial function,and maintaining the balance of extracellular matrix synthesis and catabolism.

BACKGROUND:Apoptosis plays an important role in secondary brain injury.Therefore,to explore the pathophysiological mechanism of promoting nerve cell survival after traumatic brain injury provides a new direction and theoretical basis for the prevention and treatment of traumatic brain injury. OBJECTIVE:To explore the expression changes of Lnx1 molecule in mammalian cortical neurons after brain injury and the possible mechanism involved in secondary brain injury. METHODS:Eighty adult SD rats were divided into 20 male and 20 female mice in sham operation group and 20 male and 20 female mice in traumatic brain injury group.The traumatic brain injury rat model was established by heavy falling method.At 6,12,24,48,and 72 hours after brain injury,the expression of related molecules in damaged cortical neurons was analyzed by RT-qPCR,western blot assay,and immunofluorescence staining. RESULTS AND CONCLUSION:(1)The brain tissue of traumatic brain injury group was bleeding and obvious tissue injury could be observed.Water content of brain tissue increased after traumatic brain injury.(2)Compared with the sham operation group,the expression of Lnx1 in cortical neurons after traumatic brain injury increased significantly at 24 hours after injury.(3)After traumatic brain injury,the expression of PBK and BCR protein decreased,and the pro-survival factor ctgf increased.(4)These findings suggest that after traumatic brain injury,the expression of Lnx1 is up-regulated in neurons,which may be due to the decrease of the expression of its target molecules PBK and BCR,and further promote the expression of living factor ctgf,which has a protective effect on the damaged neurons.

BACKGROUND:Restoring the normal level of reactive oxygen species and mitochondrial function of nucleus pulposus cells and inhibiting apoptosis of nucleus pulposus cells are key targets for delaying intervertebral disc degeneration.Prussian blue nanoparticles have peroxidase-like activity,which can effectively remove reactive oxygen species in the pathological microenvironment and protect nucleus pulposus cells from oxidative stress damage.OBJECTIVE:To investigate the biological functions and mechanisms of Prussian blue nanoparticles in delaying nucleus pulposus degeneration in rats.METHODS:Prussian blue nanoparticles were prepared by hydrothermal method,and their micromorphology and particle size were characterized.Prussian blue nanoparticles with different mass concentrations(20,40,60,80,and 100 μg/mL)were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.Cell proliferation was detected by CCK-8 assay after 24 hours.60 μg/mL Prussian blue nanoparticles were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.The viability of nucleus pulposus cells was observed by live-dead staining after 1 and 3 days.Passage 2 SD rat caudal vertebrae nucleus pulposus cells were obtained and observed for cell adhesion before being divided into three intervention groups.The control group did not receive any intervention.The lipopolysaccharide group was added with lipopolysaccharide.The lipopolysaccharide+Prussian blue nanoparticle group was added with lipopolysaccharide and 60 μg/mL Prussian blue nanoparticles.Reactive oxygen species,mitochondrial superoxide,and mitochondrial membrane potential were detected 24 hours after intervention.RT-qPCR detection and Alcian blue staining were performed 48 hours after intervention.RESULTS AND CONCLUSION:(1)Under transmission electron microscopy,Prussian blue nanoparticles were uniform nanocubes with an average particle size of 130 nm.(2)CCK-8 assay results showed that 20-60 μg/mL Prussian blue nanoparticles had no obvious cytotoxicity,and 60 μg/mL Prussian blue nanoparticles were selected for cell intervention in subsequent experiments.Live-dead staining results showed that 60 μg/mL Prussian blue nanoparticles had no effect on the viability of nucleus pulposus cells.(3)Compared with the control group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide group were increased(P＜0.01),the mitochondrial membrane potential was decreased(P＜0.01),the mRNA expressions of type Ⅱ collagen and aggrecan were decreased(P＜0.01),the mRNA expressions of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 were increased(P＜0.01),and the positive area of Alcian blue staining was reduced(P＜0.01).Compared with the lipopolysaccharide group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide+Prussian blue nanoparticle group were decreased(P＜0.01),mitochondrial membrane potential increased(P＜0.01),mRNA expression of type Ⅱ collagen and aggrecan increased(P＜0.01),mRNA expression of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 decreased(P＜0.01),and positive area of Alcian blue staining increased(P＜0.01).The results showed that Prussian blue nanoparticles delayed the degeneration of rat nucleus pulposus by reducing oxidative stress of nucleus pulposus cells,restoring mitochondrial function,and maintaining the balance of extracellular matrix synthesis and catabolism.

Objective:To investigate the clinical significance and inducing factors of abnormal intraoperative neurophysiological monitoring (IONM) signals during surgery for cervical degenerative diseases.Methods:A retrospective analysis was performed on 586 patients who underwent cervical degenerative disease surgery with IONM at the Department of Orthopedics, The First Affiliated Hospital of Soochow University, from April 2015 to April 2024. Surgical approaches included 380 anterior spinal canal decompression and fusion procedures, 154 posterior spinal canal decompression and fusion procedures (including single-door laminoplasty, total laminectomy, and hemilaminectomy), and 52 combined anterior-posterior surgeries. The multimodal IONM protocol employed transcranial electrical stimulation motor evoked potentials (TES-MEP) and cortical somatosensory evoked potentials (CSEP), combined with electromyography (EMG). Bilateral deltoid muscles, thenar/hypothenar muscles and abductor hallucis muscles were monitored in all patients. Intraoperative MEP, SEP, and EMG results were recorded to analyze the causes of abnormal signals, intraoperative response strategies, and postoperative neurological function and outcomes. Fourfold table chi-square tests were used to analyze factors possibly associated with IONM alerts.Results:Among the 586 cervical surgeries, 17 cases (2.9%) exhibited abnormal IONM signals. These included 4 cases of anterior cervical discectomy and fusion (ACDF), 4 cases of anterior cervical corpectomy and fusion (ACCF), and 2 cases of combined anterior-posterior surgeries for cervical spondylotic myelopathy; and 5 posterior surgeries and 2 anterior ACCF procedures for ossification of the posterior longitudinal ligament (OPLL). The rate of abnormal IONM signals was significantly higher in patients with maximum spinal cord compression (MSCC)>60% (5.8%, 12/208) than in those with MSCC≤60% (χ 2=9.417, P=0.002); in patients with intraoperative hypotension during posterior surgery (mean arterial pressure reduction>20% from baseline, cumulative duration>20 min), the abnormal IONM rate was 22.2% (6/27), which was significantly higher than that in patients without intraoperative hypotension (χ 2=33.542, P<0.001); in patients who underwent calcified tissue removal during anterior surgery, the abnormal IONM rate was 9.3% (5/54), which was significantly higher than that in patients without calcified tissue removal (χ 2=13.162, P=0.003). Thus, MSCC>60%, intraoperative hypotension during posterior surgery, and calcified tissue removal during anterior surgery may be inducing factors for abnormal IONM signals. Among the 17 patients with monitoring abnormalities, 8 cases showed no significant improvement after corresponding intraoperative treatments, and 7 of these 8 cases experienced varying degrees of muscle strength decline and sensory numbness immediately after surgery; 9 cases showed partial or complete recovery of signals, among which 8 cases had no new-onset neurological impairment after surgery, and 1 case developed unilateral upper limb grip strength decline. IONM demonstrated a sensitivity of 0.8750 and specificity of 0.8889. Conclusions:Multimodal IONM can detect electrophysiological abnormalities of spinal cord nerve function during cervical degenerative disease surgery, providing real-time warning of potential nerve damage during the operation. The proportion of abnormal IONM signals is relatively high in cases with MSCC>60%, intraoperative hypotension during posterior cervical surgery, or calcified tissue removal during anterior cervical surgery.

Objective This study aims to construct and identify the chimeric antigen receptor NK92 (CAR-NK92) cells targeting NKG2D ligand (NKG2DL) (secreting IL-15Ra-IL-15) and verify the killing activity of NKG2D CAR-NK92 cells against multiple myeloma cells. Methods The extracellular segment of NKG2D was employed to connect 4-1BB and CD3Z, as well as IL-15Ra-IL-15 sequence to obtain a CAR expression framework. The lentivirus was packaged and transduced into NK92 cells to obtain NKG2D CAR-NK92 cells. The proliferation of NKG2D CAR-NK92 cells was detected by CCK-8 assay, IL-15Ra secretion was detected by ELISA and killing efficiency was detected by lactate dehydrogenase (LDH) assay. The molecular markers of NKp30, NKp44, NKp46, the ratio of apoptotic cell population, CD107a, and the secretion level of granzyme B and perforin were detected using flow cytometry. In addition, the cytotoxic mechanism of NKG2D CAR-NK92 cells on the tumor was verified by measuring the degranulation ability. Moreover, after NKG2D antibody inhibited effector cells and histamine inhibited tumor cells, LDH assay was utilized to detect the effect on cell-killing efficiency. Finally, the multiple myeloma tumor xenograft model was constructed to verify its anti-tumor activity in vivo. Results Lentiviral transduction significantly increased NKG2D expression in NK92 cells. Compared with NK92 cells, the proliferation ability of NKG2D CAR-NK92 cells was weaker. The early apoptotic cell population of NKG2D CAR-NK92 cells was less, and NKG2D CAR-NK92 cells had stronger cytotoxicity to multiple myeloma cells. Additionally, IL-15Ra secretion could be detected in its culture supernatant. NKp44 protein expression in NKG2D CAR-NK92 cells was clearly increased, demonstrating an enhanced activation level. Inhibition test revealed that the cytotoxicity of CAR-NK92 cells to MHC-I chain-related protein A (MICA) and MICB-positive tumor cells was more dependent on the interaction between NKG2D CAR and NKG2DL. After stimulating NKG2D CAR-NK92 cells with tumor cells, granzyme B and perforin expression increased, and NK cells obviously upregulated CD107α. Furthermore, multiple myeloma tumor xenograft model revealed that the tumors of mice treated with NKG2D CAR-NK92 cells were significantly reduced, and the cell therapy did not sensibly affect the weight of the mice. Conclusion A type of CAR-NK92 cell targeting NKG2DL (secreting IL-15Ra-IL-15) is successfully constructed, indicating the effective killing of multiple myeloid cells.
Humans ; Mice ; Animals ; Receptors, Chimeric Antigen/genetics* ; Interleukin-15 ; NK Cell Lectin-Like Receptor Subfamily K/metabolism* ; Granzymes ; Cell Line, Tumor ; Multiple Myeloma/therapy* ; Perforin

Objective To explore the clinical efficacy and safety of off-clamping in robotic-assisted partial nephrectomy (RAPN) for the treatment of renal tumors.Methods From January 2015 to March 2017,the data of 48 patients who underwent off-clamping RAPN were reviewed retrospectively.There were 31 males and 17 females,and the mean age was 57 years (range:23-84 years).The mean tumor size was 3.1 cm (range:1.2-6.4 cm),with the upper,middle,and lower polar tumors account for 35.4％,27.1％,and 37.5％,respectively.The clinical tumor stage was T1N0M0 in all 48 cases,according to the AJCC tumor staging system for renal cancer.Results RAPNs were performed successfully in all 48 cases,without conversion to open surgery.In those patients,the application of off-clamping in robotic-assisted partial nephrectomy was performed in 44 cases.The renal artery and vein was exposed,dissected,isolated and then clamped in 4 cases due to bleeding.The mean surgical time was 85 min (range:75-185 min).The mean estimated blood loss was 134 ml (range:60-270 ml),and no blood transfusion was needed.The wound surface was closed using interrupted suture with Hem-o-lok clips securing each needle point.The mean time for renorrhaphy was 22 min (rang:11-31 min).No intraoperative severe complications such as vascular injury,trauma of abdominal organ occurred.There were 5 complications,including 2 cases of hematuria,2 cases of delayed healing of incision,and 1 case of pneumohypoderma.The pathological diagnosis included 40 cases of renal clear cell carcinoma,3 cases of papillary renal cell carcinoma,and 5 cases of angiomyolipomas.No tumor recurrence or distant metastasis was observed during the average follow-up of 17 months (range 3-27 months).Conclusions Off-clamping RAPN is safe and feasible approach to excise certain kidney tumors.It carries the benefits of less complication,quick recovery,and less ischemia reperfusion renal injury.Off-clamping RAPN would be suitable for those patients with solitary kidneys,renal insufficiency,and bilateral tumors.

Objective To summarize our experience in robotic-assisted laparoscopic surgery for adrenal diseases Methods The clinical data of 243 patients with adrenal tumor treated by robotic-assisted laparoscopic surgery from March 2010 to February 2017 were retrospectively reviewed.There were 99 men and 144 women.The mean age was 51.6 years (range, 19-84).Tumors located at left adrenal in 140 cases, right in 97 cases,and both sides in 6 cases.The average diameter was 3.32 cm (range, 0.8-12 cm).However, there were 41 cases whose tumor diameter were greater than 5 cm.Results There were 2 cases of conversion during operation, 1 case converted to open surgery and the other to the traditional laparoscope surgery.The mean operative time was 35 min (range, 20-130 min).Estimated blood loss was 80 ml (range,20-1 200 ml).Blood transfusion was needed in 6 cases.The mean postoperative hospital stay was 5d (range, 3-20 d).The pathological diagnosis included 37 cases of pheochromocytoma, 149 cases of cortical adenoma, 3 cases of cortical carcinoma, 5 cases of metastatic tumor, 9 cases of adrenal myelolipoma, 3 cases of adrenal cyst, 2 cases of bronchogenic cyst, 25 cases of adrenal nodular hyperplasia,2 cases of angiomyolipomas, 1 case of mature teratoma, 1 case of diffuse large B-Cell lymphoma, 1 case of angioma, and 4 cases of neurofibromatosis.Conclusions Robotic-assisted laparoscopic adrenalectomy was safe and effective.Robotic-assisted laparoscopic surgery has the advantages over laparoscopic surgery in treatment of complicated adrenal tumor, such as large adrenal tumors, pheochromocytoma, bilateral adrenal diseases, overweight and obese patients with adrenal diseases.

Aim To investigate the roles of chloride channels in the apoptosis and apoptotic volume de-crease (AVD)induced by adriamycin in nasopharyn-geal carcinoma CNE-2Z cells.Methods Apoptotic rates were detected by flow cytometry,and the volume changes were measured by the time-lapse live cell ima-ging technique.The patch clamp technique was used to record whole-cell chloride currents.Results Adria-mycin induced apoptosis of CNE-2Z cells.An early ap-optotic volume decrease was observed in the cell trea-ted with adriamycin.The cell volume was decreased by about 10% in 2 h.Adriamycin activated a chloride current which showed outward rectification.The chlo-ride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB ) could inhibit the adriamycin-in-duced chloride currents,apoptosis and prevent cell shrinkage.Conclusions Our findings suggest that ad-riamycin causes cell apoptosis by activation of chloride channels.Chloride channels may be involved in the apoptosis and apoptotic volume decrease induced by adriamycin in CNE-2Z cells.

OBJECTIVETo probe into and improve the diagnosis and treatment of tuberculosis (TB) in the male genital system.

METHODSFifteen patients with the male genital system TB established by microbiological and pathological examinations were retrospectively reviewed.

RESULTSThe male genital system TB was diagnosed by history, physical examination, semen analysis, TB culture and biopsy. Treatment consisted of antituberculous chemotherapy for all the patients before and/or after operation, including 5 cases of epididymectomy and 6 cases of orchi-epididymectomy.

CONCLUSIONThe male genital system TB tends to be atypical and semen polymerase chain reaction of TB (TB-PCR) may provide a new specific means for diagnosis in case of clinical suspicion. Surgical approaches should be cautiously adopted, especially for young patients.

Adult ; Aged ; Antitubercular Agents ; therapeutic use ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Retrospective Studies ; Tuberculosis, Male Genital ; diagnosis ; therapy