ObjectiveTo systematically characterize the in vitro chemical constituents of Entada phaseoloides and its absorbed prototype compounds and metabolites in a neuropathic pain （NP） model， thereby elucidating the potential pharmacological material basis underlying its analgesic activity. MethodsUltra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry （UHPLC-Q-Orbitrap HRMS） was used to analyze the constituents of the 80% methanol extract of E. phaseoloides. Thirty-six mice were divided into a blank group， low-， medium-， and high-dose E. phaseoloides extract groups （25， 50， and 100 mg·kg^-1， respectively）， an aspirin group （400 mg·kg^-1）， and a morphine group （10 mg·kg^-1）， with six mice in each group. The tail-flick threshold was measured. Twenty-four mice were randomly divided into a blank group and low-， medium-， and high-dose E. phaseoloides extract groups （25， 50， and 100 mg·kg^-1， respectively）， with six mice in each group， and motor coordination was evaluated. A spinal nerve ligation （SNL） model was established in mice. Twenty-four mice were randomly divided into a sham-operated （Sham） group， an SNL group， an SNL + E. phaseoloides extract group （750 mg·kg^-1）， and an SNL + pregabalin group （20 mg·kg^-1）， with 6 mice in each group. Mechanical allodynia and thermal hyperalgesia were evaluated. Behavioral assessments were combined to determine the optimal onset time of the analgesic effect， and plasma samples were collected at this time point. Absorbed components and metabolites were systematically identified based on accurate mass， MS/MS fragmentation patterns， retention time， and comparisons with literature data and reference standards. Meanwhile， network pharmacology， target similarity analysis， and molecular docking validation were integrated to evaluate the potential effects of absorbed components on disease intervention. ResultsCompared with the blank group， the high-dose E. phaseoloides extract group and morphine group showed significantly increased pain response thresholds （P<0.01）， without affecting motor coordination in normal mice. Compared with the Sham group， the SNL group exhibited significantly decreased thresholds for mechanical allodynia and thermal hyperalgesia （P<0.01）. Compared with the SNL group， the SNL + E. phaseoloides extract group and SNL + pregabalin group showed significantly increased thresholds for mechanical allodynia and thermal hyperalgesia （P<0.01）. The analgesic effect of E. phaseoloides was most significant at 2 h after administration. A total of 70 constituents were identified in vitro， and 22 prototype compounds and 29 metabolites were detected in plasma. The targets of the absorbed components were mainly enriched in biological processes associated with pain signal perception and transmission and showed a high degree of overlap with disease genes related to neuropathic pain. In addition， multiple components exhibited strong binding energies with core pain-related targets. Based on integrated analysis of in vitro content， plasma exposure levels， and target associations， ketanthamide A-β-D-pyranoside and ketengzide were identified as the most representative pharmacologically active substances responsible for the analgesic effect. ConclusionThis study systematically elucidates the material basis of E. phaseoloides intervention in neuropathic pain from three aspects， including chemical constituent characteristics， in vivo behavioral characteristics， and potential targets， providing important theoretical foundations and experimental support for further studies on its pharmacological mechanism and analgesic drug development.

Purpose To explore the characteristics of gray matter(GM)volume changes in migraine patients using 7T MRI and voxel-based morphometry(VBM).Materials and Methods This prospective study enrolled 30 migraine patients and 41 age-and gender-matched healthy controls from Beijing Tiantan Hospital,Capital Medical University between November 2023 and November 2024.All participants underwent 7T MRI with 3D T1-weighted magnetization-prepared two rapid gradient-echo(MP2RAGE)sequences for structural brain imaging.VBM analysis was performed to quantify GM,white matter,cerebrospinal fluid and total brain volumes,followed by calculations of their relative percentages.The difference in GM volume between the two groups was compared to identify brain regions with characteristic GM volume changes in migraine patients.And the correlation between these characteristic GM volume alterations and clinical scales was analyzed.Results Migraine patients exhibited significantly lower total GM volume compared to healthy controls(t=2.096,P=0.040),while no group differences were observed in white matter or cerebrospinal fluid volumes(t=0.980,0.151;P=0.330,0.880).VBM analysis revealed reduced GM volume in the left orbitofrontal cortex(t=4.301,P=0.022),left hippocampus(t=5.226,P=0.006)and left parahippocampal gyrus(t=3.960,P=0.040)in the migraine group.Negative correlations were identified between:left orbitofrontal cortex GM volume and headache duration(r=-0.506,P=0.008),left hippocampal GM volume and patient health questionnaire-9 scores(r=-0.620,P=0.003),and left parahippocampal GM volume and visual analogue scale scores(r=-0.449,P=0.019).Conclusion VBM analysis based on 7T MP2RAGE data demonstrates characteristic GM volume reductions in the left orbitofrontal cortex,left hippocampus and left parahippocampal gyrus in migraine patients,with these structural alterations significantly correlate with depressive symptoms and headache burden.The observed microstructural abnormalities may reflect underlying pathophysiological mechanisms related to pain processing,emotional regulation and long-term disease burden in migraine.

BACKGROUND:There is a central sensitization mechanism for lumbar disc herniation-specific pain sensitization,and it is unclear whether cyclic RNAs in the paraventricular nucleus brain region are involved in lumbar disc herniation-specific pain sensitization.OBJECTIVE:To investigate the mechanism by which cyclic RNA circ05188 in the paraventricular nucleus of rats with lumbar disc herniation regulates the participation of microRNAs in pain sensitization.METHODS:Fifty-two Sprague-Dawley rats were randomly divided into a sham operation group,a lumbar disc herniation group,a knockdown control group and a circ05188 knockdown group,with 13 rats in each group.Except for the sham operation group,the lumbar disc herniation model was established in the other three groups.On the 7th day after modeling,the hypothalamic paraventricular nucleus of the knockdown control group and circ05188 knockdown group were injected with siRNA-NC and siRNA-circ05188,respectively,at 1 day intervals,for a total of three injections.On the 7th day after modeling,high-throughput sequencing technology was used to detect the expression of differential circRNAs in the hypothalamic paraventricular nucleus of rats in the sham operation group and the lumbar disc herniation group,and RT-qPCR was used to detect the expression of circ05188 and miR-199a-5p in the hypothalamic paraventricular nucleus.On the 3rd,7th,14th,21st,and 28th days after modeling,we detected the mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot in the modeled side of rats in the sham operation group and the lumbar disc herniation group.On the 3rd day after siRNA injection,immunofluorescence staining was used to detect the expression of c-Fos in the hypothalamic paraventricular nucleus of rats in the knockdown control group and circ05188 knockdown group;RT-qPCR was used to detect the expression of circ05188 and miR-199a-5p in the hypothalamic paraventricular nucleus of rats in the knockdown control group and circ05188 knockdown group;on the 5th day after siRNA injection,mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot were detected in the knockdown control group and circ05188 knockdown group.Bioinformatics analysis and dual luciferase reporter elucidated the underlying molecular mechanism of circ05188 in pain sensitization after lumbar disc herniation.RESULTS AND CONCLUSION:High-throughput sequencing results showed that circRNAs were differentially expressed in the paraventricular nucleus brain region of rats with lumbar disc herniation.The expression of circ05188 was higher in the lumbar disc herniation group than in the sham operation group(P<0.05),the expression of miR-199a-5p was lower than in the sham operation group(P<0.05),and the mechanical and thermal mechanical paw withdrawal thresholds of the hindfoot were lower than those of the sham operation group at 3,7,14,and 21 days after modeling.RT-qPCR results showed that compared with the knockdown control group,the expression of c-Fos and circ05188 was lower in the circ05188 knockdown group(P<0.05),while the expression of miR-199a-5p was higher(P<0.05).The mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot in the circ05188 knockdown group were higher than those in the knockdown control group at 1,2,and 3 days after injection.Bioinformatics analysis and dual luciferase reporter assay results showed that miR-199a-5p had a binding site with circ05188 and circ05188/miR-199a-5p competitive endogenous RNA axis.To conclude,lumbar disc herniation induces an increase in circ05188 expression in the paraventricular nucleus of the hypothalamus,which produces central sensitization through the inhibition of miR-199a-5p and ultimately triggers neuropathic pain.

Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.

Purpose To explore the characteristics of gray matter(GM)volume changes in migraine patients using 7T MRI and voxel-based morphometry(VBM).Materials and Methods This prospective study enrolled 30 migraine patients and 41 age-and gender-matched healthy controls from Beijing Tiantan Hospital,Capital Medical University between November 2023 and November 2024.All participants underwent 7T MRI with 3D T1-weighted magnetization-prepared two rapid gradient-echo(MP2RAGE)sequences for structural brain imaging.VBM analysis was performed to quantify GM,white matter,cerebrospinal fluid and total brain volumes,followed by calculations of their relative percentages.The difference in GM volume between the two groups was compared to identify brain regions with characteristic GM volume changes in migraine patients.And the correlation between these characteristic GM volume alterations and clinical scales was analyzed.Results Migraine patients exhibited significantly lower total GM volume compared to healthy controls(t=2.096,P=0.040),while no group differences were observed in white matter or cerebrospinal fluid volumes(t=0.980,0.151;P=0.330,0.880).VBM analysis revealed reduced GM volume in the left orbitofrontal cortex(t=4.301,P=0.022),left hippocampus(t=5.226,P=0.006)and left parahippocampal gyrus(t=3.960,P=0.040)in the migraine group.Negative correlations were identified between:left orbitofrontal cortex GM volume and headache duration(r=-0.506,P=0.008),left hippocampal GM volume and patient health questionnaire-9 scores(r=-0.620,P=0.003),and left parahippocampal GM volume and visual analogue scale scores(r=-0.449,P=0.019).Conclusion VBM analysis based on 7T MP2RAGE data demonstrates characteristic GM volume reductions in the left orbitofrontal cortex,left hippocampus and left parahippocampal gyrus in migraine patients,with these structural alterations significantly correlate with depressive symptoms and headache burden.The observed microstructural abnormalities may reflect underlying pathophysiological mechanisms related to pain processing,emotional regulation and long-term disease burden in migraine.

BACKGROUND:There is a central sensitization mechanism for lumbar disc herniation-specific pain sensitization,and it is unclear whether cyclic RNAs in the paraventricular nucleus brain region are involved in lumbar disc herniation-specific pain sensitization.OBJECTIVE:To investigate the mechanism by which cyclic RNA circ05188 in the paraventricular nucleus of rats with lumbar disc herniation regulates the participation of microRNAs in pain sensitization.METHODS:Fifty-two Sprague-Dawley rats were randomly divided into a sham operation group,a lumbar disc herniation group,a knockdown control group and a circ05188 knockdown group,with 13 rats in each group.Except for the sham operation group,the lumbar disc herniation model was established in the other three groups.On the 7th day after modeling,the hypothalamic paraventricular nucleus of the knockdown control group and circ05188 knockdown group were injected with siRNA-NC and siRNA-circ05188,respectively,at 1 day intervals,for a total of three injections.On the 7th day after modeling,high-throughput sequencing technology was used to detect the expression of differential circRNAs in the hypothalamic paraventricular nucleus of rats in the sham operation group and the lumbar disc herniation group,and RT-qPCR was used to detect the expression of circ05188 and miR-199a-5p in the hypothalamic paraventricular nucleus.On the 3rd,7th,14th,21st,and 28th days after modeling,we detected the mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot in the modeled side of rats in the sham operation group and the lumbar disc herniation group.On the 3rd day after siRNA injection,immunofluorescence staining was used to detect the expression of c-Fos in the hypothalamic paraventricular nucleus of rats in the knockdown control group and circ05188 knockdown group;RT-qPCR was used to detect the expression of circ05188 and miR-199a-5p in the hypothalamic paraventricular nucleus of rats in the knockdown control group and circ05188 knockdown group;on the 5th day after siRNA injection,mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot were detected in the knockdown control group and circ05188 knockdown group.Bioinformatics analysis and dual luciferase reporter elucidated the underlying molecular mechanism of circ05188 in pain sensitization after lumbar disc herniation.RESULTS AND CONCLUSION:High-throughput sequencing results showed that circRNAs were differentially expressed in the paraventricular nucleus brain region of rats with lumbar disc herniation.The expression of circ05188 was higher in the lumbar disc herniation group than in the sham operation group(P<0.05),the expression of miR-199a-5p was lower than in the sham operation group(P<0.05),and the mechanical and thermal mechanical paw withdrawal thresholds of the hindfoot were lower than those of the sham operation group at 3,7,14,and 21 days after modeling.RT-qPCR results showed that compared with the knockdown control group,the expression of c-Fos and circ05188 was lower in the circ05188 knockdown group(P<0.05),while the expression of miR-199a-5p was higher(P<0.05).The mechanical and thermal mechanical paw withdrawal thresholds of the rat's hindfoot in the circ05188 knockdown group were higher than those in the knockdown control group at 1,2,and 3 days after injection.Bioinformatics analysis and dual luciferase reporter assay results showed that miR-199a-5p had a binding site with circ05188 and circ05188/miR-199a-5p competitive endogenous RNA axis.To conclude,lumbar disc herniation induces an increase in circ05188 expression in the paraventricular nucleus of the hypothalamus,which produces central sensitization through the inhibition of miR-199a-5p and ultimately triggers neuropathic pain.

Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.

BACKGROUND:With the rapid development of minimally invasive spinal surgery and enhanced recovery after surgery,endoscopic intervertebral fusion techniques have gradually emerged and been widely used in clinical practice in recent years. OBJECTIVE:To analyze the early clinical efficacy of uniaxial spinal endoscopic intervertebral fusion combined with posterior percutaneous pedicle screw fixation in the treatment of lumbar degenerative diseases. METHODS:135 patients with lumbar degenerative diseases treated by uniaxial spinal endoscopic intervertebral fusion combined with posterior percutaneous pedicle screw fixation in the Suining Central Hospital from October 2020 to December 2021 were enrolled in this study.There were 59 males and 76 females,aged 47-79 years.The lower limb and lumbar pain was evaluated by visual analog scale and lumbar function was assessed by Oswestry disability index before the operation,1 week,1,and 6 months after the operation,and at the end of follow-up.The overall pain recovery of patients was evaluated by the scoring criteria for low back pain surgery of Spine Group of Chinese Orthopedic Association and the lumbar physiological curvature and intervertebral fusion were evaluated on lumbar lateral X-ray preoperatively and at the end of follow-up. RESULTS AND CONCLUSION:(1)The 135 patients were followed up for(17.8±3.0)months after surgery.There was 1 case of endplate injury,1 case of cerebrospinal fluid leakage,1 case of nerve root injury,1 case of intervertebral cage subsidence and displacement,1 case of chronic infection,and 1 case of pedicle screw rupture.The complication rate was 5.2%.(2)The lumbar visual analog scale score and Oswestry disability index significantly decreased in the waist and lower limbs at various time points postoperatively compared with those preoperatively in 135 patients(P<0.05).The scoring criteria for low back pain surgery of the Spine Group of the Chinese Orthopedic Association were significantly better at the last follow-up than that preoperatively in 135 patients(P<0.05).(3)At the last follow-up,there was no significant difference in physiological curvature of lumbar vertebra as compared with that preoperatively in 135 patients(P>0.05),with a fusion rate of 95.8%.(4)It is concluded that uniaxial spinal endoscopic intervertebral fusion combined with posterior percutaneous pedicle screw fixation in the treatment of lumbar degenerative diseases has shown satisfactory early clinical results and is a highly safe minimally invasive spinal surgery mode.

Objective:To investigate the clinical effects of chidamide combined with linperlisib in treatment of recurrent angioimmunoblastic T-cell lymphoma (AITL) after autologous stem cell transplantation (ASCT).Methods:The clinical data of 1 patient with recurrent AITL receiving treatment of chidamide combined with linperlisib after ASCT in Sun Yat-Sen University Cancer Center in March, 2021 were retrospectively analyzed, and the related literatures were reviewed.Results:The 55- year-old male patient was presented with fatigue, night sweats and emaciation. According to the results of pathology, immunohistochemistry and imaging after admission, this patient was diagnosed as AITL. After treatment with venbutuximab + CDP (cyclophosphamide + doxorubicin + prednisone) regimen, the patient achieved complete remission, and then ASCT was performed. After the transplatation, programmed death receptor 1 inhibitor was maintained for 4 courses of treatment. It recurred 16 months after ASCT, and the patient achieved partial remission after 1 course of chidamide combined with linperlisib, and achieved complete remission after 2 courses of treatment. Later, recheck of hepatitis B virus showed a quantitative increase after the self withdrawal of anti hepatitis B drugs, and then the primary treatment was suspended. The last follow-up time was March, 2024, and recheck results of whole body enhanced magnetic resonance imaging indicated recurrence.Conclusions:The combination of chidamide and linperlisib is effective and safe in treatment of recurrent ATIL after ASCT.

Objective:To explore the independent risk factors of portal vein thrombosis (PVT) in liver cirrhosis, and to establish and evaluate a risk prediction model for PVT in patients with cirrhosis.Methods:A total of 295 cases of cirrhosis hospitalized in Renmin Hospital of Wuhan University from December 2019 to October 2021 were divided into a modeling set ( n=207) and an internal validation set ( n=88) by the random number table. In addition, patients with cirrhosis hospitalized in Yichang Central People's Hospital, Wuhan Puren Hospital, No.2 People's Hospital of Fuyang City and People's Hospital of China Three Gorges University during the same period were collected as an external validation set ( n=92). The modeling set was divided into PVT group ( n=56) and non-PVT group ( n=151). Univariate analysis was used to preliminarily screen the related indicators of PVT, and then multivariate logistic regression analysis with forward stepwise regression was used to determine independent risk factors for PVT. A nomogram prediction model was constructed based on the independent risk factors obtained. The internal and external validation set were used to verify the predictive ability of the model. Distinction degree was used to evaluate the ability of the model to distinguish patients with or without PVT. Hosmer-Lemeshow goodness-of-fit test was used to evaluate the consistency between predicted risk and the actual risk of the model. Results:Univariate analysis showed that smoking, history of splenectomy, trans-jugular intrahepatic portosystemic shunt (TIPS), gastrointestinal bleeding and endoscopic variceal treatment, and levels of hemoglobin, alanine aminotransferase, aspartate aminotransferase and D-dimer were significantly different between the PVT group and the non-PVT group ( P<0.05). Multivariate logistic regression analysis found that smoking ( P=0.020, OR=31.21, 95% CI: 1.71-569.40), levels of D-dimer ( P=0.003, OR=1.12, 95% CI: 1.04-1.20) and hemoglobin ( P=0.039, OR=0.99, 95% CI: 0.97-1.00), history of TIPS ( P=0.011, OR=18.04, 95% CI: 1.92-169.90) and endoscopic variceal treatment ( P=0.001, OR=3.21, 95% CI: 1.59-6.50) were independent risk factors for PVT in patients with liver cirrhosis. Receiver operator characteristic (ROC) curve analysis showed that the area under the ROC curve (AUC) for the internal validation set was 0.802 (95% CI: 0.709-0.895) ( P<0.001), and the AUC for the external validation set was 0.811 (95% CI: 0.722-0.900) ( P<0.001). Both AUC were larger than 0.75. The calibration curve of Hosmer-Lemeshow goodness-of-fit test showed that the P values of both internal validation set ( χ2=3.602, P=0.891) and the external validation set ( χ2=11.025, P=0.200) were larger than 0.05. Conclusion:Smoking, history of TIPS or endoscopic variceal treatment, levels of D-dimer and hemoglobin are independent risk factors for PVT in patients with liver cirrhosis. The prediction nomogram model based on the above factors has strong predictive ability.