Neuropathic pain is a chronic condition caused by damage or dysfunction in the nervous system. While the spleen may influence neuropathic pain, its role has been poorly understood. This study demonstrates that the spleen plays a crucial role in regulating neuropathic pain through the bed nucleus of the stria terminalis (BNST) - paraventricular nucleus of the hypothalamus (PVN) neural circuit in a chronic constriction injury (CCI) mouse model. Splenectomy, splenic denervation, or splenic sympathectomy significantly increased the mechanical withdrawal threshold (MWT) and reduced macrophage infiltration in the dorsal root ganglia (DRG) of CCI mice. Pseudorabies virus injections into the spleen revealed connections to the BNST and PVN in the brain. Chemogenetic inhibition of the BNST-PVN circuit increased macrophage infiltration in the DRG and decreased the MWT; these effects were reversed by splenectomy, splenic denervation, or sympathectomy. These findings underscore the critical role of the spleen, regulated by the BNST-PVN circuit, in neuropathic pain.
Animals ; Neuralgia/pathology* ; Septal Nuclei/physiopathology* ; Male ; Spleen/physiopathology* ; Paraventricular Hypothalamic Nucleus/physiopathology* ; Mice, Inbred C57BL ; Splenectomy ; Mice ; Neural Pathways/physiopathology* ; Disease Models, Animal ; Ganglia, Spinal/physiopathology* ; Sympathectomy ; Macrophages

OBJECTIVE: To explore the association between blood glucose trajectories within 7 days of intensive care unit (ICU) admission and mortality in patients with sepsis-associated acute respiratory distress syndrome (ARDS). METHODS: Based on the MIMIC-IV database, sepsis-associated ARDS patients with daily blood glucose monitoring data within 7 days of ICU admission were selected. Blood glucose trajectories were analyzed using group-based trajectory modeling (GBTM), and the optimal number of groups was determined based on the minimum Akaike information criterion (AIC), Bayesian information criterion (BIC), average posterior probability (AvePP), odds of correct classification (OCC), and proportion of group membership (Prop). Baseline characteristics including demographics, comorbidities, severity scores, vital signs, laboratory indicators within the first 24 hours of ICU admission, and treatments were collected. Kaplan-Meier survival curves were used to compare 28-day and 1-year survival across trajectory groups. Multivariate Logistic regression was performed to evaluate the associations between glucose trajectory groups and in-hospital mortality, ICU mortality. The incidence of hypoglycemia within 7 days in the ICU was analyzed among different groups. RESULTS: A total of 3 869 patients with sepsis-associated ARDS were included, with a median age of 63.52 (52.13, 73.54) years; 59.6% (2 304/3 869) were male. Based on glucose levels within 7 days, patients were categorized into three groups: persistent hyperglycemia group (glucose maintained at 10.6-13.1 mmol/L, n = 894), moderate glucose group (7.8-8.9 mmol/L, n = 1 452), and low-normal glucose group (6.1-7.0 mmol/L, n = 1 523). There were statistically significant differences in 28-day mortality and 1-year mortality among low-normal glucose group, moderate glucose group, and persistent hyperglycemia group [28-day mortality: 11.42% (174/1 523), 19.83% (288/1 452), 25.50% (228/894), χ ² = 82.545, P < 0.001; 1-year mortality: 23.31% (355/1 523), 33.75% (490/1 452), 39.49% (353/894), χ ² = 77.376, P < 0.001]. Kaplan-Meier analysis showed that higher glucose trajectories were associated with significantly lower 28-day and 1-year cumulative survival rates (Log-rank test: χ ² were 83.221 and 85.022, both P < 0.001). There were statistically significant differences in in-hospital mortality and ICU mortality among the low-normal glucose group, moderate glucose group, and persistent hyperglycemia group [in-hospital mortality: 9.65% (147/1 523), 19.70% (286/1 452), 24.50% (219/894), χ ² = 102.020, P < 0.001; ICU mortality: 7.22% (110/1 523), 16.05% (233/1 452), 20.13% (180/894), χ ² = 93.050, P < 0.001]. Logistic regression confirmed that, using the persistent hyperglycemia group as the reference, the low-normal glucose group had significantly lower risks of in-hospital mortality and ICU mortality after multiple factor adjustment. Although the moderate glucose group showed a trend toward lower mortality, the differences were not statistically significant. Using the moderate glucose group as a reference, the low-normal glucose group had 43.1% lower in-hospital mortality [odds ratio (OR) = 0.569, 95% confidence interval (95%CI) was 0.445-0.726, P < 0.001] and 42.0% lower ICU mortality (OR = 0.580, 95%CI was 0.439-0.762, P < 0.001). There was no statistically significant difference in the incidence of hypoglycemia within 7 days of ICU admission among low-normal glucose group, moderate glucose group, and persistent hyperglycemia group [2.82% (43/1 523), 2.69% (39/1 452), 3.02% (27/894), χ ² = 0.226, P = 0.893]. CONCLUSIONS Blood glucose trajectories during ICU stay are closely associated with prognosis in patients with sepsis-associated ARDS. Persistent hyperglycemia (10.6-13.1 mmol/L) is linked to significantly higher short- and long-term mortality.
Humans ; Respiratory Distress Syndrome/etiology* ; Sepsis/blood* ; Intensive Care Units ; Male ; Female ; Middle Aged ; Blood Glucose/metabolism* ; Hospital Mortality ; Aged

OBJECTIVE: To explore the genetic basis of a patient with suspected Oculocutaneous albinism (OCA). METHODS: An OCA patient presented at the West China Second Hospital of Sichuan University and his mother were selected as the study subjects. Peripheral blood samples were collected for the extraction of, genomic DNA, and whole exome sequencing (WES) was carried out. Candidate variants were verified through specific primer amplification, Sanger sequencing, and agarose gel electrophoresis. Bioinformatic analysis and pathogenicity rating were conducted on the candidate variants. This study has been approved by the Medical Ethics Committee of West China Second Hospital (No. 2024-228). RESULTS: Genetic testing revealed that the patient had harbored variants in exon 1 of the TYR gene, including a c.157G>T (p.G53C) missense variant and a c.609dup (p.A204fs) frameshifting variant. Specific primer amplification and Sanger sequencing, combined with agarose gel electrophoresis, confirmed that these are compound heterozygous variants. Based on the guidelines from the ACMG, the c.157G>T was rated as likely pathogenic, and c.609dup was rated as pathogenic. Alphafold3 predicted that the variant proteins had significant structural changes. CONCLUSION The patient was diagnosed with OCA due to compound heterozygous variants of the TYR gene. Discovery of the c.609dup variant has enriched the mutational spectrum of OCA and provided a basis for genetic counseling and prenatal diagnosis for this patient.
Humans ; Albinism, Oculocutaneous/enzymology* ; Male ; Female ; Monophenol Monooxygenase/chemistry* ; Base Sequence ; Mutation, Missense

Objective:To explore the genetic basis of a patient with suspected Oculocutaneous albinism (OCA).Methods:An OCA patient presented at the West China Second Hospital of Sichuan University and his mother were selected as the study subjects. Peripheral blood samples were collected for the extraction of genomic DNA, and whole exome sequencing (WES) was carried out. Candidate variants were verified through specific primer amplification, Sanger sequencing, and agarose gel electrophoresis. Bioinformatic analysis and pathogenicity rating were conducted on the candidate variants. This study has been approved by the Medical Ethics Committee of West China Second Hospital (No. 2024-228).Results:Genetic testing revealed that the patient had harbored variants in exon 1 of the TYR gene, including a c. 157G>T (p.G53C) missense variant and a c. 609dup (p.A204fs) frameshifting variant. Specific primer amplification and Sanger sequencing, combined with agarose gel electrophoresis, confirmed that these are compound heterozygous variants. Based on the guidelines from the ACMG, the c. 157G>T was rated as likely pathogenic, and c. 609dup was rated as pathogenic. Alphafold3 predicted that the variant proteins had significant structural changes. Conclusion:The patient was diagnosed with OCA due to compound heterozygous variants of the TYR gene. Discovery of the c. 609dup variant has enriched the mutational spectrum of OCA and provided a basis for genetic counseling and prenatal diagnosis for this patient.

Objective:To evaluate the role of splenic sympathetic nerve-regulated infiltration and polarization of dorsal root ganglion (DRG) macrophages in diabetic neuropathic pain (DNP) in mice.Methods:Forty-eight specific pathogen-free male C57BL/6 mice, aged 6 weeks, weighing 20-22 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (Con group), DNP group, DNP plus sham operation group (DNP+ Sham group), and diabetes mellitus induced by DNP plus splenic sympathetic denervation group (DNP+ SS group). In DNP+ SS group, the splenic sympathetic denervation procedures were performed using 6-hydroxydopamine solution, while a solvent of 0.2% ascorbic acid saline solution was used as a substitute for 6-hydroxydopamine solution in DNP+ Sham group. After a two-week recovery, the diabetes mellitus was induced by intraperitoneal injection of streptozotocin 120 mg/kg in mice at 8 weeks of age. The mechanical paw withdrawal threshold (MWT) were measured on day 1 before developing the model and on days 7, 14, 21 and 28 after developing the model. After the last behavioral testing, the DRG was taken after anesthesia for determination of the expression of the macrophage marker ionized calcium-binding adaptor molecule 1(Iba1), calcitonin gene-related peptide (CGRP), and tumor necrosis factor-α (TNF-α) (by immunofluorescence) and expression of M1 phenotype markers (CD16, TNF-α, inducible nitric oxide synthase [iNOS]) and M2 phenotype markers (interleukin-10 [IL-10], transforming growth factor-β1 [TGF-β1], and CD206) mRNA (using quantitative real-time polymerase chain reaction). Results:Compared with Con group, the MWT was significantly decreased on days 14, 21 and 28 after developing the model, the expression of CGRP and TNF-α in the DRG was up-regulated, the count of Iba1-positive cells was increased, the expression of CD16, TNF-α and iNOS mRNA was up-regulated ( P<0.05), and no significant change was found in the expression of IL-10, TGF-β1 and CD206 in DNP group ( P>0.05). Compared with DNP group and DNP+ Sham group, the MWT was significantly increased on days 14, 21 and 28 after developing the model, the expression of CGRP and TNF-α in the DRG was down-regulated, the count of Iba1-positive cells was decreased, the expression of CD16, TNF-α and iNOS mRNA was down-regulated, and the expression of IL-10, TGF-β1 and CD206 mRNA was up-regulated in DNP+ SS group ( P<0.05), and no significant change was found in the aforementioned parameters at each time point in DNP and DNP+ Sham groups ( P>0.05). Conclusions:Activation of splenic sympathetic nerve can promote the infiltration and polarization of DRG macrophages, thus participating in the process of diabetic neuropathic pain in mice.

Objective:To evaluate the effect of dexmedetomidine on the viability of dopaminergic neurons in the ventral tegmental area (VTA) of morphine-addicted mice.Methods:Experiment Ⅰ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using the random number table method: normal saline group (NS group), dexmedetomidine 50 μg/kg group (DEX50 group), and dexmedetomidine 100 μg/kg group (DEX100 group). A morphine addiction model was established by intraperitoneal injection of increasing doses of morphine (10, 20, 30, 40, 50 and 50 mg/kg) for 6 consecutive days in mice. After the successful establishment of the model, dexmedetomidine 50 and 100 μg/kg were intraperitoneally injected for 14 consecutive days in group DEX50 and group DEX100 respectively, while normal saline was given instead in group C. The conditioned place preference (CPP) experiment was conducted every other day. Experiment Ⅱ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) by the random number table method: control group (C group), morphine group (Mor group) and dexmedetomidine 50 μg/kg group (DEX50 group). Normal saline was intraperitoneally injected for 10 consecutive days in group C. Morphine with increasing doses was intraperitoneally injected for 6 days, and then normal saline was intraperitoneally injected for 4 consecutive days in group Mor. Morphine with increasing doses was intraperitoneally injected for 6 days, and then dexmedetomidine 50 μg/kg was intraperitoneally injected for 4 consecutive days in group DEX50. The mice were anesthetized at 90 min after the last intraperitoneal injection, brain tissues were harvested, and the corresponding brain slices of the VTA were selected for c-Fos immunofluorescence staining. Experiment Ⅲ Ten dopamine transporter-Cre recombinase mice were divided into 2 groups ( n=5 each) by the random number table method: morphine group (Mor group) and morphine+ dexmedetomidine 50 μg/kg group (Mor+ DEX group). Stereotaxic viral injection was performed in the brain. rAAV-EF1α-DIO-GCaMP6s was injected into the VTA and an optical fiber was implanted. Three weeks later, a morphine addiction model was established based on Experiment Ⅰ for the CPP experiment, morphine was intraperitoneally injected in group Mor, and morphine and dexmedetomidine were intraperitoneally injected in group Mor+ DEX. The viral fluorescence signals were recorded at 5 min before and 20 min after the drug administration in the three groups. Results:Experiment Ⅰ There was no statistically significant difference in the CPP scores after developing the morphine addiction model among the three groups ( P>0.05). Compared with group NS, the CPP scores were significantly decreased at 4-14 days of the continuous administration in group DEX50 and group DEX100 ( P<0.05). Experiment Ⅱ Compared with group C, the number of c-Fos positive cells in the VTA was significantly increased in group Mor ( P<0.05). Compared with group Mor, the number of c-Fos positive cells in the VTA was significantly decreased in group DEX ( P<0.05). Experiment Ⅲ Compared with that before administration, the calcium signals of dopaminergic neurons in the VTA were significantly enhanced in group Mor ( P<0.05), and no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA in group Mor+ DEX ( P>0.05). Compared with group Mor, no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA before drug administration ( P>0.05), and the calcium signals of dopaminergic neurons in the VTA were significantly weakened after administration in group Mor+ DEX ( P<0.05). Conclusions:The mechanism by which dexmedetomidine promotes the extinction of morphine addiction is related to the inhibition of the viability of dopaminergic neurons in the VTA of mice.

Objective:To evaluate the role of splenic sympathetic nerve-regulated infiltration and polarization of dorsal root ganglion (DRG) macrophages in diabetic neuropathic pain (DNP) in mice.Methods:Forty-eight specific pathogen-free male C57BL/6 mice, aged 6 weeks, weighing 20-22 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (Con group), DNP group, DNP plus sham operation group (DNP+ Sham group), and diabetes mellitus induced by DNP plus splenic sympathetic denervation group (DNP+ SS group). In DNP+ SS group, the splenic sympathetic denervation procedures were performed using 6-hydroxydopamine solution, while a solvent of 0.2% ascorbic acid saline solution was used as a substitute for 6-hydroxydopamine solution in DNP+ Sham group. After a two-week recovery, the diabetes mellitus was induced by intraperitoneal injection of streptozotocin 120 mg/kg in mice at 8 weeks of age. The mechanical paw withdrawal threshold (MWT) were measured on day 1 before developing the model and on days 7, 14, 21 and 28 after developing the model. After the last behavioral testing, the DRG was taken after anesthesia for determination of the expression of the macrophage marker ionized calcium-binding adaptor molecule 1(Iba1), calcitonin gene-related peptide (CGRP), and tumor necrosis factor-α (TNF-α) (by immunofluorescence) and expression of M1 phenotype markers (CD16, TNF-α, inducible nitric oxide synthase [iNOS]) and M2 phenotype markers (interleukin-10 [IL-10], transforming growth factor-β1 [TGF-β1], and CD206) mRNA (using quantitative real-time polymerase chain reaction). Results:Compared with Con group, the MWT was significantly decreased on days 14, 21 and 28 after developing the model, the expression of CGRP and TNF-α in the DRG was up-regulated, the count of Iba1-positive cells was increased, the expression of CD16, TNF-α and iNOS mRNA was up-regulated ( P<0.05), and no significant change was found in the expression of IL-10, TGF-β1 and CD206 in DNP group ( P>0.05). Compared with DNP group and DNP+ Sham group, the MWT was significantly increased on days 14, 21 and 28 after developing the model, the expression of CGRP and TNF-α in the DRG was down-regulated, the count of Iba1-positive cells was decreased, the expression of CD16, TNF-α and iNOS mRNA was down-regulated, and the expression of IL-10, TGF-β1 and CD206 mRNA was up-regulated in DNP+ SS group ( P<0.05), and no significant change was found in the aforementioned parameters at each time point in DNP and DNP+ Sham groups ( P>0.05). Conclusions:Activation of splenic sympathetic nerve can promote the infiltration and polarization of DRG macrophages, thus participating in the process of diabetic neuropathic pain in mice.

Objective:To evaluate the effect of dexmedetomidine on the viability of dopaminergic neurons in the ventral tegmental area (VTA) of morphine-addicted mice.Methods:Experiment Ⅰ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using the random number table method: normal saline group (NS group), dexmedetomidine 50 μg/kg group (DEX50 group), and dexmedetomidine 100 μg/kg group (DEX100 group). A morphine addiction model was established by intraperitoneal injection of increasing doses of morphine (10, 20, 30, 40, 50 and 50 mg/kg) for 6 consecutive days in mice. After the successful establishment of the model, dexmedetomidine 50 and 100 μg/kg were intraperitoneally injected for 14 consecutive days in group DEX50 and group DEX100 respectively, while normal saline was given instead in group C. The conditioned place preference (CPP) experiment was conducted every other day. Experiment Ⅱ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) by the random number table method: control group (C group), morphine group (Mor group) and dexmedetomidine 50 μg/kg group (DEX50 group). Normal saline was intraperitoneally injected for 10 consecutive days in group C. Morphine with increasing doses was intraperitoneally injected for 6 days, and then normal saline was intraperitoneally injected for 4 consecutive days in group Mor. Morphine with increasing doses was intraperitoneally injected for 6 days, and then dexmedetomidine 50 μg/kg was intraperitoneally injected for 4 consecutive days in group DEX50. The mice were anesthetized at 90 min after the last intraperitoneal injection, brain tissues were harvested, and the corresponding brain slices of the VTA were selected for c-Fos immunofluorescence staining. Experiment Ⅲ Ten dopamine transporter-Cre recombinase mice were divided into 2 groups ( n=5 each) by the random number table method: morphine group (Mor group) and morphine+ dexmedetomidine 50 μg/kg group (Mor+ DEX group). Stereotaxic viral injection was performed in the brain. rAAV-EF1α-DIO-GCaMP6s was injected into the VTA and an optical fiber was implanted. Three weeks later, a morphine addiction model was established based on Experiment Ⅰ for the CPP experiment, morphine was intraperitoneally injected in group Mor, and morphine and dexmedetomidine were intraperitoneally injected in group Mor+ DEX. The viral fluorescence signals were recorded at 5 min before and 20 min after the drug administration in the three groups. Results:Experiment Ⅰ There was no statistically significant difference in the CPP scores after developing the morphine addiction model among the three groups ( P>0.05). Compared with group NS, the CPP scores were significantly decreased at 4-14 days of the continuous administration in group DEX50 and group DEX100 ( P<0.05). Experiment Ⅱ Compared with group C, the number of c-Fos positive cells in the VTA was significantly increased in group Mor ( P<0.05). Compared with group Mor, the number of c-Fos positive cells in the VTA was significantly decreased in group DEX ( P<0.05). Experiment Ⅲ Compared with that before administration, the calcium signals of dopaminergic neurons in the VTA were significantly enhanced in group Mor ( P<0.05), and no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA in group Mor+ DEX ( P>0.05). Compared with group Mor, no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA before drug administration ( P>0.05), and the calcium signals of dopaminergic neurons in the VTA were significantly weakened after administration in group Mor+ DEX ( P<0.05). Conclusions:The mechanism by which dexmedetomidine promotes the extinction of morphine addiction is related to the inhibition of the viability of dopaminergic neurons in the VTA of mice.

Objective:To explore the prognosis of patients with gastric cancer peritoneal metastasis (PM) receiving programmed cell death-1 (PD-1) antibody therapy, and investigate the biomarkers that affect the prognosis of anti-PD-1 therapy.Methods:This restrospecific study collected the clinic-pathological data of 56 patients with peritoneal metastasis of gastric cancer who received first-line treatment in the Nanjing Drum Town Hospital from March 2020 to September 2023, among which 41 had received anti-PD-1 immunotherapy and 15 hadn't. The relationship between overall survival (OS) and anti-PD-1 immunotherapy was evaluated by Kaplan-Meier analysis. The relationship between baseline peripheral blood indicators and treatment response of patients with anti-PD-1 treatment was analyzed using unpaired t-test. Subsequently, the Cox proportional risk regression model was used to explore the clinical prognostic factors that may affect anti-PD-1 immunotherapy by univariate and multivariate analysis. The clinical prognostic factors included baseline data and baseline peripheral blood indexes such as anti-PD-1 treatment lines, Eastern Cooperative Oncology Group performance status (ECOG PS), combined positive score (CPS), expression of human epidermal growth factor receptor 2 (Her-2), EBER status, pathological types, other metastatic lesions, ascites content before immunotherapy, with or without abdominal drainage during anti-PD-1 treatment, blood lipid indicators, inflammatory indicators, and tumor indicators. Results:Kaplan-Meier survival statistics showed similar OS (15.9 vs. 15.2 months, P=0.600) in patients with anti-PD-1 therapy compared to those without anti-PD-1 therapy. Patients with baseline high-density lipoprotein (HDL) ≥0.97 mmol/L ( n=22) demonstrated a significantly longer median OS compared to those with HDL＜0.97 mmol/L (15.2 vs. 13.5 months; P=0.018). Similarly, the cohort with apolipoprotein A1 (ApoA1) levels ≥0.86 g/L ( n=21) showed superior survival outcomes, with a median OS of 17.7 months versus 12.3 months in the ApoA1＜0.86 g/L group ( n=20; P=0.006). In contrast, elevated baseline alpha-fetoprotein (AFP) levels ( n=2) were associated with markedly reduced survival (median OS: 5.7 vs. 15.2 months in normal AFP group, n=37; P=0.005). Notably, elevated pretreatment ApoA1 levels correlated with enhanced immunotherapy response ( P=0.017). Multivariate Cox regression analysis revealed that ApoA1 deficiency (≥0.86 g/L) independently predicted better OS following PD-1 antibody therapy ( HR=0.35, 95% CI: 0.12-0.98, P=0.046) in gastric cancer patients with PM. Conclusions:In our study, it is first proposed that ApoA1 could be a significant predictor of the survival advantages of immunotherapy in gastric cancer patients with PM.

Objective:To explore the genetic basis of a patient with suspected Oculocutaneous albinism (OCA).Methods:An OCA patient presented at the West China Second Hospital of Sichuan University and his mother were selected as the study subjects. Peripheral blood samples were collected for the extraction of genomic DNA, and whole exome sequencing (WES) was carried out. Candidate variants were verified through specific primer amplification, Sanger sequencing, and agarose gel electrophoresis. Bioinformatic analysis and pathogenicity rating were conducted on the candidate variants. This study has been approved by the Medical Ethics Committee of West China Second Hospital (No. 2024-228).Results:Genetic testing revealed that the patient had harbored variants in exon 1 of the TYR gene, including a c. 157G>T (p.G53C) missense variant and a c. 609dup (p.A204fs) frameshifting variant. Specific primer amplification and Sanger sequencing, combined with agarose gel electrophoresis, confirmed that these are compound heterozygous variants. Based on the guidelines from the ACMG, the c. 157G>T was rated as likely pathogenic, and c. 609dup was rated as pathogenic. Alphafold3 predicted that the variant proteins had significant structural changes. Conclusion:The patient was diagnosed with OCA due to compound heterozygous variants of the TYR gene. Discovery of the c. 609dup variant has enriched the mutational spectrum of OCA and provided a basis for genetic counseling and prenatal diagnosis for this patient.