Immune thrombocytopenia (ITP) is a hemorrhagic autoimmune disease characterized by antibody-mediated platelet injury. ITP has complicated immunopathological mechanisms that need further elucidation. It is well known that the costimulatory molecules OX40 ligand (OX40L) and OX40 play essential roles in the immunological mechanisms of autoimmune diseases. Previously, we discovered that the expression of OX40L and OX40 is significantly increased in the peripheral blood mononuclear cells (PBMCs) of ITP patients. In our present study, OX40L-induced follicular helper T (Tfh) cells exhibited an activated phenotype with elevated expression of inducible T-cell costimulator (ICOS), programmed cell death protein-1 (PD-1), and cluster of differentiation 40 ligand (CD40L) in vitro. Moreover, aberrant OX40L‒OX40 expression might promote the Tfh1-to-Tfh2 shift in vivo, inducing the generation of autoantibodies by enhancing the helper function of Tfh cells for B lymphocytes in a mouse model, which might accelerate the progression of ITP. Additionally, signal transduction through the OX40L‒OX40 axis might be related to the activation of tumor necrosis factor receptor-associated factor (TRAF)‒nuclear factor-κB (NF-κB) and Janus kinase (JAK)‒signal transducer and activator of transcription (STAT) signaling pathways. Overall, OX40L‒OX40 signaling is proposed as a potential novel therapeutic target for ITP.
Animals ; OX40 Ligand/physiology* ; Purpura, Thrombocytopenic, Idiopathic/immunology* ; Cell Differentiation ; Mice ; T-Lymphocytes, Helper-Inducer/cytology* ; T Follicular Helper Cells/cytology* ; Signal Transduction ; Receptors, OX40 ; Mice, Inbred C57BL ; Humans ; Female

Extensive evidence has demonstrated that glucagon-like peptide-1 receptor agonists (GLP-1RAs) can ameliorate obesity. Our previous studies revealed that (Ex-4)₂-Fc, a long-acting GLP-1RA we developed, depends on the leptin pathway to treat obesity. However, the mechanisms linking (Ex-4)₂-Fc and leptin resistance remain largely unclear. To address this question, we explored the mechanism of GLP-1RAs from the perspective of the gut microbiota, as increasing evidence indicates an important link between the gut microbiota and obesity. This study aimed to explore the potential role of the gut microbiota in the treatment of GLP-1RAs. We found that (Ex-4)₂-Fc treatment reshaped obesity-induced gut microbiota disturbances and substantially increased the abundance of Akkermansia muciniphila (Am). In addition, (Ex-4)₂-Fc did not respond well in antibiotic-treated (ATB) Obese mice. Subsequent studies have shown that this defect can be overcome by gavage with Am. In addition, we found that Am enhanced (Ex-4)₂-Fc therapy by producing the metabolite inosine. Inosine regulates the macrophage adenosine A2A receptor (A2A) pathway to indirectly reduce leptin levels in adipocytes Thus, elucidating the role of metabolites in regulating the leptin pathway will provide new insights into GLP-1RAs therapy and may lead to more effective strategies for guiding the clinical use of antidiabetic agents.

Objective:To explore the short-term clinical effects of deep cervical lymph node-venous anastomosis in the treatment of Alzheimer’s disease (AD).Methods:A prospective exploratory study was conducted on the treatment of AD patients using the cervical deep lymph node-venous anastomosis technique in Scar and Wound Treatment Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, from September to October 2024. The patients underwent high-frequency ultrasound to locate deep cervical lymph nodes and the external jugular vein. Under general anesthesia, bilateral deep cervical lymph node-venous anastomoses were performed. Indocyanine green (ICG) lymphography was conducted via subcutaneous injection behind the ear to visualize lymph nodes in levels Ⅱ and Ⅲ. After making a skin incision along the posterior margin of the sternocleidomastoid muscle, the external jugular vein, internal jugular veins, and associated lymph nodes were exposed. Adjacent veins were selected for anastomosis of lymph node. Using microsurgical techniques, end-to-side or end-to-end anastomosis was completed for lymph nodes in levels Ⅱ and Ⅲ. Preoperative assessments included the mini-mental state examination (MMSE, a higher score indicates better cognitive function), Alzheimer’s disease assessment scale-cognitive subscale (ADAS-Cog, a higher score indicates greater impairment of cognitive function), Alzheimer’s disease cooperative study scale for activities of daily living (ADCS-ADL, a higher score indicates better ability to perform daily activity), and neuropsychiatric inventory (NPI, a higher score indicates more severe behavioral and emotional symptom). Postoperative follow-up included the same scales to observe changes in cognitive function, activities of daily living, and emotional communication.Results:Four patients (1 male, 3 females, aged 58-79 years) with AD were included. All were diagnosed based on cerebrospinal fluid biomarkers. All patients successfully underwent bilateral deep cervical lymph node-venous anastomoses. On average, 4.3 (2-7 per person) anastomoses were performed per patient. Surgical procedures lasted an average of 6.5 h (5.5-8.5 h) with minimal blood loss (less than 50 ml). Patients resumed normal activity within 6 hours postoperatively and were discharged after an average of 4.1 d (3.5-5.0 d). Postoperative complications included one case each of aspiration pneumonia, lower limb venous thrombosis, and transient delirium, all of whom resolved without long-term effects. Clinical symptoms, including memory decline, mood swings, and anxiety, showed varying degrees of improvement. Patients reported enhanced quality of life, emotional stability, and social engagement, confirming the procedure’s safety and potential cognitive benefits. At one month postoperatively, the MMSE scores of the four patients increased by an average of 0.8 points compared to preoperative levels. Additionally, the two patients who completed the ADAS-Cog assessments showed a decrease in their scores (reduced by 1.0 points and 11.3 points, respectively, compared to preoperative scores), indicating a certain degree of improvement in cognitive function during this period. The ADCS-ADL and NPI scores of four patients varied significantly, without showing any clear pattern.Conclusion:Lymphovenous anastomosis of the deep cervical lymph node-venous anastomosis may provide a new surgical intervention approach for AD, but further large-scale studies and long-term follow-up are needed to validate its safety and effectiveness.

Objective:To explore the diagnostic value of transvaginal color Doppler sonography (TVCDS) combined with serum allograft inflammatory factor-1 (AIF-1) and pyruvate kinase M2 type (PKM2) in cervical cancer.Methods:From Jan. 2022 to Jun. 2024, 168 patients with early cervical cancer who visited Taizhou Central Hospital were regarded as the cervical cancer group, and 96 patients with cervical precancerous lesions were regarded as the precancerous lesion group. All patients underwent TVCDS examination to obtain parameters such as peak systolic velocity (PSV), end diastolic velocity (EDV), and resistance index (RI). ELISA method was used to detect serum AIF-1 and PKM2 levels. Kappa test was used to analyze the consistency between different diagnostic methods and pathological results. ROC curve was used to analyze the diagnostic value of serum AIF-1 and PKM2 levels for cervical cancer.Results:The cervical cancer group had manifestly higher proportions of irregular morphology, unclear boundaries, low echo ratio, and levels of PSV, EDV, AIF-1, and PKM2 than precancerous lesion group, and manifestly lower RI than precancerous lesion group ( P<0.05). The consistency analysis between TVCDS diagnosis of cervical cancer and pathological results showed a Kappa value of 0.674 ( P<0.05). The serum levels of AIF-1 and PKM2 in cervical cancer patients with a maximum tumor diameter of ≥ 4 cm, low differentiation, and vaginal infiltration were manifestly higher than those in patients with a maximum tumor diameter of < 4 cm, medium/high differentiation, and non vaginal infiltration ( P<0.05). The AUC of AIF-1 and PKM2 in the diagnosis of cervical cancer was 0.824 and 0.816, respectively, with diagnostic thresholds of 81.21 ng/L and 29.26 ng/mL. The consistency analysis of TVCDS combined with serum AIF-1 and PKM2 in the diagnosis of cervical cancer and pathological results showed a Kappa value of 0.918 ( P<0.05). The negative predictive value, accuracy, and sensitivity of TVCDS combined with serum AIF-1 and PKM2 in the diagnosis of cervical cancer were higher than those of each indicator alone, and the positive predictive value and specificity were higher than those of AIF-1 alone ( P<0.05) . Conclusion:TVCDS combined with serum AIF-1 and PKM2 has high accuracy and sensitivity in the diagnosis of cervical cancer, and their combination can serve as a potential method for clinical diagnosis of cervical cancer.

Objective To investigate the effect of Lactobacillus plantarum LB12 on kidney calcium oxalate stones in rats.Methods A total of 24 male SD rats aged 6～8 weeks were randomly divided into four groups:control group,model group,LB12 group,and LB12 intervention group,with 6 rats in each group.The control group and LB12 group had free access to water,and were gavaged with 2 mL of normal saline in the morning.In the evening,the control group received 2 mL of normal saline,while the LB12 group received 2 mL of Lactobacillus plantarum LB12 suspension.The model group and LB12 intervention group were given 1%ethylene glycol in drink-ing water and 2 mL of 2%ammonium chloride solution via gavage in the morning.In the evening,both groups received 2 mL of normal saline and 2 mL of Lactobacillus plantarum LB12 suspension,respectively.After the experiment,the body weight change trend of the rats was recorded,and the levels of urinary calcium,urinary oxalate,serum creatinine,blood urea nitrogen,renal SOD,and MDA were measured.Additionally,H&E staining,VK staining,immunohistochemistry,ROS fluorescence analysis,and Western blot(WB)were performed on kidney tissues.Results Lactobacillus plantarum LB12 demonstrated a 12.34%oxalate degradation rate in vitro.Further animal experiments indicated that Lactobacillus plantarum LB12 alleviated the weight loss trend in the model rats and reduced the levels of urinary calcium,urinary oxalate,serum creatinine,urea nitrogen,and MDA,while increasing SOD levels(P<0.05).Histopathological analysis revealed that the LB12 intervention group had signifi-cantly fewer renal crystals compared to the model group,with lower expressions of IL-6 and ROS(P<0.01).Western blot analysis demonstrated that Lactobacillus plantarum LB12 inhibited the expression of TLR4,NF-κB,and OPN proteins(P<0.01).Conclusions Lactobacillus plantarum LB12 can effectively alleviate the oxidative stress damage caused by calcium oxalate stones to the kidneys and reduce kidney crystal formation in rats.Further-more,it inhibits the expression of TLR4/NF-κB and OPN,making it a potential therapeutic approach for the prevention and treatment of kidney calcium oxalate stones.

Objective:To explore the diagnostic value of transvaginal color Doppler sonography (TVCDS) combined with serum allograft inflammatory factor-1 (AIF-1) and pyruvate kinase M2 type (PKM2) in cervical cancer.Methods:From Jan. 2022 to Jun. 2024, 168 patients with early cervical cancer who visited Taizhou Central Hospital were regarded as the cervical cancer group, and 96 patients with cervical precancerous lesions were regarded as the precancerous lesion group. All patients underwent TVCDS examination to obtain parameters such as peak systolic velocity (PSV), end diastolic velocity (EDV), and resistance index (RI). ELISA method was used to detect serum AIF-1 and PKM2 levels. Kappa test was used to analyze the consistency between different diagnostic methods and pathological results. ROC curve was used to analyze the diagnostic value of serum AIF-1 and PKM2 levels for cervical cancer.Results:The cervical cancer group had manifestly higher proportions of irregular morphology, unclear boundaries, low echo ratio, and levels of PSV, EDV, AIF-1, and PKM2 than precancerous lesion group, and manifestly lower RI than precancerous lesion group ( P<0.05). The consistency analysis between TVCDS diagnosis of cervical cancer and pathological results showed a Kappa value of 0.674 ( P<0.05). The serum levels of AIF-1 and PKM2 in cervical cancer patients with a maximum tumor diameter of ≥ 4 cm, low differentiation, and vaginal infiltration were manifestly higher than those in patients with a maximum tumor diameter of < 4 cm, medium/high differentiation, and non vaginal infiltration ( P<0.05). The AUC of AIF-1 and PKM2 in the diagnosis of cervical cancer was 0.824 and 0.816, respectively, with diagnostic thresholds of 81.21 ng/L and 29.26 ng/mL. The consistency analysis of TVCDS combined with serum AIF-1 and PKM2 in the diagnosis of cervical cancer and pathological results showed a Kappa value of 0.918 ( P<0.05). The negative predictive value, accuracy, and sensitivity of TVCDS combined with serum AIF-1 and PKM2 in the diagnosis of cervical cancer were higher than those of each indicator alone, and the positive predictive value and specificity were higher than those of AIF-1 alone ( P<0.05) . Conclusion:TVCDS combined with serum AIF-1 and PKM2 has high accuracy and sensitivity in the diagnosis of cervical cancer, and their combination can serve as a potential method for clinical diagnosis of cervical cancer.

Objective To investigate the effect of Lactobacillus plantarum LB12 on kidney calcium oxalate stones in rats.Methods A total of 24 male SD rats aged 6～8 weeks were randomly divided into four groups:control group,model group,LB12 group,and LB12 intervention group,with 6 rats in each group.The control group and LB12 group had free access to water,and were gavaged with 2 mL of normal saline in the morning.In the evening,the control group received 2 mL of normal saline,while the LB12 group received 2 mL of Lactobacillus plantarum LB12 suspension.The model group and LB12 intervention group were given 1%ethylene glycol in drink-ing water and 2 mL of 2%ammonium chloride solution via gavage in the morning.In the evening,both groups received 2 mL of normal saline and 2 mL of Lactobacillus plantarum LB12 suspension,respectively.After the experiment,the body weight change trend of the rats was recorded,and the levels of urinary calcium,urinary oxalate,serum creatinine,blood urea nitrogen,renal SOD,and MDA were measured.Additionally,H&E staining,VK staining,immunohistochemistry,ROS fluorescence analysis,and Western blot(WB)were performed on kidney tissues.Results Lactobacillus plantarum LB12 demonstrated a 12.34%oxalate degradation rate in vitro.Further animal experiments indicated that Lactobacillus plantarum LB12 alleviated the weight loss trend in the model rats and reduced the levels of urinary calcium,urinary oxalate,serum creatinine,urea nitrogen,and MDA,while increasing SOD levels(P<0.05).Histopathological analysis revealed that the LB12 intervention group had signifi-cantly fewer renal crystals compared to the model group,with lower expressions of IL-6 and ROS(P<0.01).Western blot analysis demonstrated that Lactobacillus plantarum LB12 inhibited the expression of TLR4,NF-κB,and OPN proteins(P<0.01).Conclusions Lactobacillus plantarum LB12 can effectively alleviate the oxidative stress damage caused by calcium oxalate stones to the kidneys and reduce kidney crystal formation in rats.Further-more,it inhibits the expression of TLR4/NF-κB and OPN,making it a potential therapeutic approach for the prevention and treatment of kidney calcium oxalate stones.

Objective:To explore the short-term clinical effects of deep cervical lymph node-venous anastomosis in the treatment of Alzheimer’s disease (AD).Methods:A prospective exploratory study was conducted on the treatment of AD patients using the cervical deep lymph node-venous anastomosis technique in Scar and Wound Treatment Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, from September to October 2024. The patients underwent high-frequency ultrasound to locate deep cervical lymph nodes and the external jugular vein. Under general anesthesia, bilateral deep cervical lymph node-venous anastomoses were performed. Indocyanine green (ICG) lymphography was conducted via subcutaneous injection behind the ear to visualize lymph nodes in levels Ⅱ and Ⅲ. After making a skin incision along the posterior margin of the sternocleidomastoid muscle, the external jugular vein, internal jugular veins, and associated lymph nodes were exposed. Adjacent veins were selected for anastomosis of lymph node. Using microsurgical techniques, end-to-side or end-to-end anastomosis was completed for lymph nodes in levels Ⅱ and Ⅲ. Preoperative assessments included the mini-mental state examination (MMSE, a higher score indicates better cognitive function), Alzheimer’s disease assessment scale-cognitive subscale (ADAS-Cog, a higher score indicates greater impairment of cognitive function), Alzheimer’s disease cooperative study scale for activities of daily living (ADCS-ADL, a higher score indicates better ability to perform daily activity), and neuropsychiatric inventory (NPI, a higher score indicates more severe behavioral and emotional symptom). Postoperative follow-up included the same scales to observe changes in cognitive function, activities of daily living, and emotional communication.Results:Four patients (1 male, 3 females, aged 58-79 years) with AD were included. All were diagnosed based on cerebrospinal fluid biomarkers. All patients successfully underwent bilateral deep cervical lymph node-venous anastomoses. On average, 4.3 (2-7 per person) anastomoses were performed per patient. Surgical procedures lasted an average of 6.5 h (5.5-8.5 h) with minimal blood loss (less than 50 ml). Patients resumed normal activity within 6 hours postoperatively and were discharged after an average of 4.1 d (3.5-5.0 d). Postoperative complications included one case each of aspiration pneumonia, lower limb venous thrombosis, and transient delirium, all of whom resolved without long-term effects. Clinical symptoms, including memory decline, mood swings, and anxiety, showed varying degrees of improvement. Patients reported enhanced quality of life, emotional stability, and social engagement, confirming the procedure’s safety and potential cognitive benefits. At one month postoperatively, the MMSE scores of the four patients increased by an average of 0.8 points compared to preoperative levels. Additionally, the two patients who completed the ADAS-Cog assessments showed a decrease in their scores (reduced by 1.0 points and 11.3 points, respectively, compared to preoperative scores), indicating a certain degree of improvement in cognitive function during this period. The ADCS-ADL and NPI scores of four patients varied significantly, without showing any clear pattern.Conclusion:Lymphovenous anastomosis of the deep cervical lymph node-venous anastomosis may provide a new surgical intervention approach for AD, but further large-scale studies and long-term follow-up are needed to validate its safety and effectiveness.

Objective:To evaluate the relationship between the mechanism of preoperative sleep deprivation aggravating postoperative cognitive dysfunction and theta oscillations in the hippocampal CA1 region in aged mice.Methods:Twenty-four SPF male C57BL/6J mice, aged 18 months, weighing 20-30 g, were divided into 3 groups ( n=8 each) by a random number table method: normal control group (group C), operation group (group S) and preoperative sleep deprivation + operation group (group SD+ S). The sleep deprivation model was established using the sleep deprivation rod method, and the sleep deprivation time was 24 h before operation. The sleep deprivation rod was set to rotate continuously at 6 rpm/min, and the direction was randomly reversed to force the mice to exercise to achieve the purpose of sleep deprivation. Group C received no treatment. Group S underwent exploratory laparotomy under isoflurane anesthesia. After sleep deprivation for 24 h, SD+ S group underwent exploratory laparotomy under isoflurane anesthesia. Y-maze and Morris water maze tests were used to evaluate postoperative cognitive function. The theta oscillations in the hippocampal CA1 region were observed and recorded by EEG electrodes in vivo. The animals were then sacrificed and brain tissues were obtained for determination of the density of dendritic spines in the hippocampal CA1 region. Results:Compared with group C, the number of crossing the original platform was significantly decreased, and the time of staying at the original platform quadrant and time of exploring the novel arm were shortened, and the number of entries into the novel arm was reduced, the density of dendritic spines in the hippocampal CA1 region was reduced, and the percentage of theta oscillation power was decreased during the Y maze test in group S ( P<0.05). Compared with S group, the number of crossing the original platform was significantly decreased, and the time of staying at the original platform quadrant and time of exploring the novel arm were shortened, and the number of entries into the novel arm was reduced, the density of dendritic spines in the hippocampal CA1 region was reduced, and the percentage of theta oscillation power was decreased during the Y maze test in SD+ S group ( P<0.01). Conclusions:The mechanisms by which preoperative sleep deprivation exacerbates postoperative cognitive dysfunction may be related to reduction in theta oscillations in the hippocampal CA1 region of aged mice.

Objective:To explore the effects of different feeding patterns on weight gain and feeding intolerance in hospitalized premature infants.Methods:The retrospective cohort study design was adopted. From April 2019 to October 2020, 155 premature infants admitted to the Neonatal Intensive Care Unit of China-Japan Friendship Hospital were selected as the study subject by convenience sampling. Premature infants were divided into a mixed feeding group ( n=47) and a formula milk feeding group ( n=108) based on their feeding patterns. The observation indicators included weight gain, feeding intolerance, infectious diseases, nutrition and other related indicators. Results:The weight change value and the weight change rate during hospitalization in the mixed feeding group were higher than those in the formula milk feeding group, with statistically significant differences ( P<0.05) . In terms of the total milk volume on the day of discharge, the mixed feeding group were higher than the formula milk feeding group, and the differences were statistically significant ( P<0.05) . During hospitalization, the number of new infectious diseases in the mixed feeding group was less than that in the formula milk feeding group, with a statistically significant difference ( P<0.05) . There was no statistically significant difference in the incidence of feeding intolerance between the two groups of premature infants ( P>0.05) , The incidence of vomiting in the formula milk feeding group was lower than that in the mixed feeding group, with a statistically significant difference ( P<0.05) . Conclusions:Mixed feeding is more conducive to promoting the weight gain of premature infants during hospitalization and reducing the occurrence of infectious diseases than formula milk feeding. The impact of two feeding patterns on feeding intolerance in hospitalized premature infants needs further comparative analysis.