OBJECTIVES: To investigate the role of a neural pathway from oxytocin (OXT) neurons in the paraventricular hypothalamic nucleus (PVN) to γ-aminobutyric acid (GABA) neurons (GABAergic neurons) in the trigeminal nucleus caudalis (TNC) in regulating pain sensitization in a mouse model of chronic migraine and to explore the underlying mechanisms. METHODS: A chronic migraine mouse model was established by intraperitoneal injection of nitroglycerin (NTG, 1 mg/mL, 10 mg/kg) on days 1, 3, 5, 7, and 9. The study consisted of four parts: PartⅠ: 24 male wild-type C57BL/6J mice were divided into four groups (n=6 in each), receiving single or repeated injection of NTG or saline, respectively. Immunofluorescence was used to detect c-Fos and OXT expression in the PVN. Part Ⅱ: 6 male OXT-Cre transgenic C57BL/6J mice were used for anterograde monosynaptic tracing combined with RNAscope and immunofluorescence to identify neural projections from PVN OXT neurons to TNC GABAergic neurons. Part Ⅲ: 30 male OXT-Cre transgenic C57BL/6J mice were bilaterally injected Cre-dependent chemogenetic activation virus into the PVN. These mice were randomly divided into five groups, with six mice in each group. Mice in the clozapine N-oxide (CNO) group and the control group were intra-peritoneally injected with 0.1 mg/mL of CNO solution (1 mg/kg) and the same volume of isotonic normal saline, respectively. 3 hours after the injection, the brain tissues were harvest and c-Fos immunofluorescence staining was performed to verify the efficiency of chemogenetic activation virus. Mice in the model control group and the CNO activated model group were subjected to chronic migraine modeling, with bilateral TNC injection of isotonic normal saline and CNO, respectively, on day 10. The mice in the negative control group were bilaterally intra-TNC injected with isotonic normal saline. After 30 minutes, the Von-Frey filament and acetone tests were used to assess the mechanical pain threshold and cold pain response time in the periorbital region of the mice in these three groups. Part Ⅳ: 24 male OXT-Cre transgenic C57BL/6J mice were bilaterally injected with the Cre-dependent chemogenetic activation virus into the PVN. These mice were randomly divided into four groups, with six mice in each group. Mice in the model control group, the CNO activated model group and the atosiban group were subjected to chronic migraine modeling. On day 10, mice in the negative control group and the model control group were intraperitoneally injected with isotonic normal saline, while mice in the CNO activated model group and the atosiban group were intraperitoneally injected with CNO. After 15 minutes, mice in the atosiban group were bilaterally intra-TNC injected with atosiban, while mice in other three groups were bilaterally intra-TNC injected with isotonic normal saline containing 1% dimethyl sulfoxide. After 15 minutes, the Von-Frey filament and acetone tests were used to assess the mechanical pain threshold and cold pain response time in the periorbital region of the mice. The GABA content in the bilateral TNC was detected by high-performance liquid chromatography (HPLC). RESULTS: Mice with chronic migraine models exhibited reduced periorbital mechanical pain thresholds and increased periorbital cold pain reaction time, accompanied by an increase in both the number of c-Fos⁺ neurons and the percentage of c-Fos⁺ OXT neurons in the PVN (all P<0.05). The anterograde tracing virus and RNAscope combined with immunofluorescence staining showed that PVN OXT neurons projected to TNC GABAergic neurons. Immuno-fluorescence staining demonstrated that compared with the control group, the percentage of c-Fos⁺ OXT neurons in the PVN of CNO group increased (P<0.05). In bilateral intra-TNC drug administration experiments, compared with the model control group, the periorbital mechanical pain threshold increased, and the periorbital cold pain reaction time decreased in the CNO activated model group (both P<0.05). In intraperitoneal drug administration experiments, compared with the CNO activate model group, the periorbital mechanical pain threshold decreased, and the periorbital cold pain reaction time increased in the atosiban group (both P<0.05). HPLC analysis showed that, compared with the negative control group, the model control group and the atosiban group, GABA level of TNC in the CNO activated model group increased (all P<0.05). CONCLUSIONS PVN OXT neurons exert a descending facilitatory effect on GABAergic neurons in the TNC via OXT release, thereby ameliorating pain sensitization in chronic migraine.
Animals ; Paraventricular Hypothalamic Nucleus/physiopathology* ; Male ; Mice, Inbred C57BL ; Migraine Disorders/physiopathology* ; Mice ; GABAergic Neurons/physiology* ; Oxytocin/physiology* ; Disease Models, Animal ; Neurons/physiology* ; Mice, Transgenic ; Neural Pathways ; Chronic Disease

Objective:To investigate the perioperative safety and survival benefits of two-stage operation in the treatment of esophageal malignancies.Methods:This is a retrospective case series study. The clinical data of 70 patients who underwent two-stage operation for esophageal malignancies from January 2015 to January 2023 in the Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine were retrospectively analyzed. There were 58 males and 12 females. The age ( M(IQR)) was 61 (22) years (range: 9 to 79 years), and 51.4% (36/70) of the patients were older than 65 years. All patients underwent the first operation to remove esophageal tumor and the second operation to complete digestive tract reconstruction. The reasons for two-stage operation were summarized, complications and treatment results were recorded, and the prognosis was calculated using Kaplan-Meier curve. Results:The interval between the two stages was 49 (35) days (range: 32 to 82 days). The follow-up perrod was 28(50) months (range: 1 to 69 months). The 30-day fatality rate after the first operation was 4.3% (3/70) and the 30-day fatality rate after the second operation was 0(0/63). The total complication rate of first operation was 55.7% (39/70), the respiratory complication rate was 10.0% (7/70) and the cardiovascular complication rate was 5.7% (4/70) The incidence of anastomotic leakage was 11.1% (9/63). The overall survival rate at 2, 3 and 5 years was 35.7%, 30.0% and 15.7%, respectively.Conclusion:Two-stage operation in the treatment of complex esophageal malignancies is feasible and yields satisfaetory results.

[Objective] To explore the clinical application of cutaneous ureterostomy-flap embedding in radical cystectomy plus urinary diversion. [Methods] The clinical data of 10 patients with bladder cancer treated with this method in our hospital during Feb.and May 2023 were involved.Cutaneous ureterostomy-flap embedding was used in urinary diversion.The stoma-free rate and stenosis rate of stomas within 1 year postoperatively, differences in renal function indicators 1 day before operation and 1 year after operation, urinary diversion-related complications within 6 months postoperatively, including hydronephrosis, urinary tract infections, renal stones were analyzed. [Results] All surgeries were successfully completed.At 1 year postoperatively, renal function indicators showed no significant difference compared to preoperative levels (P>0.05). At 6 months postoperatively, 1 patient developed renal stones, successfully treated with surgery; 2 had urinary tract infection, recovered after antibiotic treatment; 2 had mild unilateral hydronephrosis, alleviated with conservative management.At 1 year postoperatively, the catheter-free rate was 80%(8/10), with no worsening of hydronephrosis or occurrence of ureteral obstruction, and the stent placement duration ranged from 97 to 211 days, average (151.63±42.47) days.The ureteral stent was not removed in 2 patients within 1 year, so the stoma stenosis rate was 20%(2/10). [Conclusion] The application of flap embedding in urinary diversion following radical cystectomy is a simple and safe procedure, with few postoperative complications, high success rate of stent removal, and overall favorable outcomes.

Objective:To investigate the perioperative safety and survival benefits of two-stage operation in the treatment of esophageal malignancies.Methods:This is a retrospective case series study. The clinical data of 70 patients who underwent two-stage operation for esophageal malignancies from January 2015 to January 2023 in the Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine were retrospectively analyzed. There were 58 males and 12 females. The age ( M(IQR)) was 61 (22) years (range: 9 to 79 years), and 51.4% (36/70) of the patients were older than 65 years. All patients underwent the first operation to remove esophageal tumor and the second operation to complete digestive tract reconstruction. The reasons for two-stage operation were summarized, complications and treatment results were recorded, and the prognosis was calculated using Kaplan-Meier curve. Results:The interval between the two stages was 49 (35) days (range: 32 to 82 days). The follow-up perrod was 28(50) months (range: 1 to 69 months). The 30-day fatality rate after the first operation was 4.3% (3/70) and the 30-day fatality rate after the second operation was 0(0/63). The total complication rate of first operation was 55.7% (39/70), the respiratory complication rate was 10.0% (7/70) and the cardiovascular complication rate was 5.7% (4/70) The incidence of anastomotic leakage was 11.1% (9/63). The overall survival rate at 2, 3 and 5 years was 35.7%, 30.0% and 15.7%, respectively.Conclusion:Two-stage operation in the treatment of complex esophageal malignancies is feasible and yields satisfaetory results.

Leptospirosis is a global natural zoonotic infectious disease.Research shows that short-chain fatty acids,metabolites of gut microbiota,are involved in host immune regulation and affect disease progression.This experiment was conducted to investigate the role of short-chain fatty acids in leptospirosis.The results showed that supplementation of short-chain fatty acid butyric acid could significantly improve the survival rate of leptospirosis in hamsters.In vitro experiments showed that butyric acid treatment inhibited the expression of Cat and Gsr genes in macrophages after infection with Leptospira,but enhanced the expression of NOX1 and NOX4 genes.At the same time,butyric acid treatment enhanced macrophage ROS levels after leptospirosis infection,and high levels of ROS enhanced the bactericidal function of macrophages.In vivo experiments al-so confirmed that butyric acid protects hamsters against acute leptospirosis by regulating ROS production.Collectively,the short-chain fatty acid butyric acid,a metabolite of gut microbiota,en-hances the bactericidal function of macrophages by regulating ROS expression,thereby protecting the host against leptospirosis.

Objective:To investigate the time characteristics from extracorporeal cardiopulmonary resuscitation (ECPR) initiation to termination.Methods:The data of ECPR patients in extracorporeal life support center of the First Affiliated Hospital of Nanjing Medical University from April 2015 to October 2023 were retrospectively analyzed. The patients were grouped by survival/death at 90 days, in-hospital/out-of-hospital cardiac arrest (IHCA/OHCA), daytime/evening initiation, and procedural/non-procedural termination. Data on age, sex, Charlson comorbidity index, interhospital transport, initial rhythm, ECPR initial blood gas pH and lactate value, no-flow time, time from cardiac arrest to extracorporeal membrane oxygenation (ECMO) initiation (CA-Pump On time), ECPR initiation/termination and ECMO treatment time, 90-day survival rate and so on were analyzed.Results:200 ECPR patients were enrolled, the cardiogenic etiologies were accounted for 70.5%, more men than women, 68 (34.0%) patients survived at 90 days, of whom 61 (89.7%) patients had good neurological outcomes. The 90-day survival group had a significantly lower of no-flow time, a higher proportion of IHCA and initial shockable heart rhythm, with a higher ECPR initial blood gas pH and a lower lactic acid value than those in the death group. 3. The no flow time in OHCA group was significantly longer than that in IHCA group, with a lower ECPR initial blood gas pH and a higher lactic acid value, 77.4% were non-procedural termination and the 90-day survival rate was 16.1%. ECPR were initiated in all time periods, IHCA-ECPR initiated at random, OHCA-ECPR were rare in the early morning, and the initiation time had no significant effect on ECPR outcomes. There were 75.5% of ECPR terminated at the daytime, 56.8% death cases were occurred within 3 days after ECPR, and 19.0% of patients in the procedural termination group died due to a combination of factors.Conclusions:ECPR had a potentially high benefit for patients with IHCA, initial shockable rhythm, and a short no-flow time. The ECPR initiation time were irregular and had no effect on ECPR outcomes. Death events tend to occur in the early days after ECPR, and ECPR terminated is mostly during the daytime working hours. The construction of full-time ECPR team should be strengthened.

Objective:To evaluate the prognostic value of coagulofibrinolytic changes in survivors of adult extracorporeal cardiopulmonary resuscitation (ECPR) in Chinese.Methods:133 cardiac arrest (CA) patients whose blood samples were collected after establishment of extracorporeal membrane oxygenation between January 2018 and April 2023 in Emergency Department of the First Affiliated Hospital of Nangjing Medical University were enrolled in this single-center study. The following were examined: platelet counts (PLT), prothrombin time (PT), activated partial thrombin time (APTT), thrombin time (TT), fibrinogen (Fib), and D-dimer (D-D). The prognostic values of these coagulation-related indicators in predicting poor outcomes (Cerebral Performance Category 3-5) were analyzed.Results:Among the 133 patients involved, 96 (72.2%) had poor outcomes after ECPR treatment. In the poor-outcome group, PLT and Fib were significantly decreased, while PT and APTT were significantly prolonged (all P < 0.05). ECPR survivors had a significantly higher risk of poor neurological outcomes in the high-level group than those in the low-level group for PT ( HR=1.87, 95% CI:1.17-2.99, P=0.009), APTT ( HR=1.95, 95% CI:1.26-3.00, P=0.003), and D-D levels ( HR=5.18, 95% CI:2.06-13.03, P<0.001). The risk of poor neurological outcomes was significantly lower in the high-level group for PLT ( HR=0.55, 95%CI:0.35-0.84, P=0.007). Conclusion:Coagulofibrinolytic changes can be promising tools to assess poor neurological outcomes in adult ECPR survivors.

Objective:To observe the effects of early energy intake and early enteral nutrition on prognosis of patients during extracorporeal membrane oxygenation (ECMO).Methods:Patients who received ECMO treatment in the emergency intensive care unit (EICU) of the Jiangsu Provincial Hospital (First Affiliated Hospital of Nanjing Medical University) from January 2021 to June 2022 were selected as subjects to summarize the early energy intake of ECMO patients. Logistic regression analysis and restricted cubic spline (RCS) analysis were used to determine the relationship between early energy intake and prognosis of ECMO patients. According to the results of RCS analysis, the patients were divided into energy-deficient group and energy-sufficient group. And according to whether early enteral nutrition (EEN) was initiated, the patients were divided into EEN group and non-EEN group. The differences of clinical outcomes between energy-deficient group and energy-sufficient group, EEN group and non-EEN group were compared.Results:There was no significant difference in age, sex, BMI, primary disease and ECMO pattern between energy-deficient group and energy-sufficient group, EEN group and non-EEN group. The ECMO conversion time (days) and hospitalization time (days) in the energy-deficient group were significantly lower than those in the energy-sufficient group, and the survival rate in the energy-deficient group was significantly lower than that in the energy-sufficient group [43.2% (19/44) vs. 66.0% (31/47), P=0.029]. Kaplan-Meier survival analysis showed that the 28-day survival rate in the energy-deficient group was significantly lower than that in the energy-sufficient group, and the risk of death was 2.595 times higher than that in the energy-sufficient group. The conversion time (days), hospital stay (days) and average daily energy intake [kcal/(kg·d)] in the EEN group were higher than those in the non-EEN group ( P<0.05), and the survival rate in the non-EEN group was significantly higher than that in the non-EEN group [66.1% (41/62) vs. 31.0% (9/29), P<0.002]. Kaplan-Meier survival analysis showed that the 28-day survival rate in the non-EEN group was significantly lower than that in the EEN group, and the risk of death was 2.981 times higher than that in the EEN group ( P<0.001). Conclusions:The energy intake of patients with ECMO above 16.94 kcal/ (kg·d) is a protective factor for prognosis. EEN helps to increase early energy intake and improve prognosis in patients during ECMO.

Objective:To investigate the effect of Early infarct growth rate(EIGR) on the prognosis of patients with acute large vessel occlusive ischemic stroke.Methods:A total of 164 patients with acute large vessel occlusive ischemic stroke were enrolled in the emergency department of the First Affiliated Hospital of Nanjing Medical University from January 1, 2020 to December 31, 2022.According to the change of the National Institutes of Health Stroke Scale (NIHSS) score at admission and 72 h after treatment, the patients were divided into good prognosis group and poor prognosis group. The basic clinical data of the two groups were observed and compared. The risk factors of poor prognosis were analyzed by univariate regression. The effect of EIGR on prognosis after age stratification was further analyzed.Results:Comparing the clinical data of the two groups, there was no difference in EIGR (mL/h) (7.67 vs. 8.24, P=0.211) between the two groups. The product between EIGR and age was included as the interaction term, and the result of the interaction term in the model was statistically significant ( OR=1.002, 95% CI: 1.000-1.003, P=0.032) .Moreover, the result was still statistically significant after adjusting for relevant variables (gender, history of hypertension, history of atrial fibrillation, history of diabetes, history of coronary heart disease, and history of stroke) ( OR=1.002, 95% CI:1.000-1.003, P=0.027). Subgroup analysis was performed according to the median age (71 years). In the elderly group, the proportion of poor prognosis was higher with fast core infarction growth rate defined by 25 mL/h and 15 mL/h ( P < 0.05).In the younger age group, there was no significant difference in the proportion of poor prognosis in the fast core infarction growth rate compared with the slow type ( P > 0.05). Conclusions:EIGR can predict the early clinical outcome early in elderly patients with large vessel occlusive ischemic stroke.