OBJECTIVE: To elucidate the specific mechanisms by which electroacupuncture (EA) alleviates anxiety and fear behaviors associated with posttraumatic stress disorder (PTSD), focusing on the role of lipocalin-2 (Lcn2). METHODS: The PTSD mouse model was subjected to single prolonged stress and shock (SPS&S), and the animals received 15 min sessions of EA at Shenmen acupoint (HT7). Behavioral tests were used to investigate the effects of EA at HT7 on anxiety and fear. Western blotting and enzyme-linked immunosorbent assay were used to quantify Lcn2 and inflammatory cytokine levels in the prefrontal cortex (PFC). Additionally, the activity of PFC neurons was evaluated by immunofluorescence and in vivo electrophysiology. RESULTS: Mice subjected to SPS&S presented increased anxiety- and fear-like behaviors. Lcn2 expression in the PFC was significantly upregulated following SPS&S, leading to increased expression of the proinflammatory cytokines tumor necrosis factor-α and interleukin-6 and suppression of PFC neuronal activity. However, EA at HT7 inhibited Lcn2 release, reducing neuroinflammation and hypoexcitability in the PFC. Lcn2 overexpression mitigated the effects of EA at HT7, resulting in anxiety- and fear-like behaviors. CONCLUSION EA at HT7 can ameliorate PTSD-associated anxiety and fear, and its mechanism of action appears to involve the inhibition of Lcn2-mediated neural activity and inflammation in the PFC. Please cite this article as: Yang YD, Zhong W, Chen M, Tang QC, Li Y, Yao LL, et al. Electroacupuncture alleviates behaviors associated with posttraumatic stress disorder by modulating lipocalin-2-mediated neuroinflammation and neuronal activity in the prefrontal cortex. J Integr Med. 2025; 23(5):537-547.
Electroacupuncture ; Stress Disorders, Post-Traumatic/metabolism* ; Animals ; Lipocalin-2/metabolism* ; Prefrontal Cortex/physiopathology* ; Male ; Mice ; Neurons/physiology* ; Disease Models, Animal ; Fear ; Behavior, Animal ; Mice, Inbred C57BL ; Neuroinflammatory Diseases/metabolism* ; Anxiety/therapy* ; Acupuncture Points

Background and Aims:Papillary thyroid carcinoma(PTC),the most common type of thyroid cancer,has been rapidly increasing in incidence worldwide,posing a serious threat to individual health and public healthcare systems.Exposure to low-to-moderate doses of ionizing radiation is more relevant to the daily lives of the general population and,therefore,raises greater public health concerns.It has also been widely recognized as a potential factor in immune system remodeling.This study was conducted to investigate the impact of low-to-moderate dose ionizing radiation on the tumor immune microenvironment of PTC,aiming to reveal the potential hazards of such radiation exposure in PTC patients.Methods:Two datasets(GSE29265 and GSE35570)containing RNA-seq data and corresponding clinical information were retrieved and downloaded from the GEO database.These datasets included thyroid cancer samples from patients exposed to ionizing radiation due to the Chernobyl disaster,as well as sporadic thyroid cancer cases.After data cleaning,merging,batch effect correction,differential gene expression analysis,functional enrichment analysis,immune cell infiltration analysis,and tumor microenvironment analysis were performed using R language.Results:In tumor samples,the radiation-exposed group exhibited significant differential gene expression compared to the sporadic group,with three genes upregulated and 27 genes downregulated.These differentially expressed genes were primarily enriched in biological functions closely related to immune responses,including chemokine activity,immune cell chemotaxis,and tumor immunity.Immune cell infiltration analysis indicated that radiation exposure had a limited impact on immune cell infiltration in normal samples.However,in tumor samples,the immune and ESTIMATE scores were significantly lower in the radiation-exposed group than in the sporadic group.Further analysis revealed that total T cells,CD4+T cells,CD8+T cells,B cells,and cytotoxic lymphocytes exhibited significantly lower infiltration levels in the tumor microenvironment of the radiation-exposed group than the sporadic group.Conclusion:Although low-to-moderate dose ionizing radiation has a relatively minor impact on normal thyroid tissue,it significantly reduces the infiltration of various immune cell subtypes in the PTC tumor microenvironment.This reduction in immune infiltration may have important implications for disease progression.

The purpose of this study is to develop a shark single domain antibody(SdAb)targeting a unique B cell epitope in the SARS-CoV-2 spike protein,and explore its role in the immunological detection targeting SARS-CoV-2 spike protein S.A u-nique peptide S9 was artificially synthesized based on the sequence of a unique B cell epitope of SARS-CoV-2 spike protein,then it was conjugated to the carrier protein KLH.It was used as an immunogen for subcutaneous injection into shark back and boosted according to the standard immunization protocol.Blood collected from shark tail vein and peripheral blood lymphocytes(PBL)were isolated.Total RNA was purified from PBL and transcribed to cDNA by reverse transcription.Shark vNAR frag-ments were amplified from cDNA templates and cloned into pComb3XSS vector to obtain phage library.A positive clone named T01 was obtained through screening the phage library by indirect ELISA.Then its gene was cloned into the expression vector pET-28a.The SdAb T01 was then prokaryotically expressed and purified,and its specific recognition of the SARS-CoV-2 spike protein S was indentified by Western-blot(WB),indirect ELISA and IF A.T01 binds well with peptide S9 at EC50 value of 2.050±0.064 nmol/L.The purified SdAb T01 was proven by WB to be able to selectively detect recombinant spike protein sub-unit 1(S1)of SARS-CoV-2,with no cross-reactive to recombinant spike protein subunit 1 of other six human coronavirus.It was showed by ELISA that SdAb T01 can sensitively detect the recombinant N terminal domain(NTD)of SARS-CoV-2 pro-tein.Moreover,it also specifically recognizes the spike protein of SARS-CoV-2 that was transiently expressed in transfected HEK293 cells by IFA.Therefore,a shark single domain antibody targeting a unique B cell epitope in the spike protein of SARS-CoV-2 was successfully developed,and has shown potential immunodiagnostic value by WB,ELISA and IFA.Thus,it provides an effective tool for unique antigen detection of SARS-CoV-2.

Background and Aims:Papillary thyroid carcinoma(PTC)is the most common endocrine malignancy in China,with cervical lymph node metastasis being a frequent and critical clinical feature that directly affects patient prognosis and recurrence risk.In recent years,with the rapid increase in the prevalence of overweight and obesity in China,the role of body mass index(BMI)in various tumors has attracted growing attention.This study aimed to investigate the association between overweight and cervical LNM in PTC,analyze sex-specific differences and influencing factors,and provide evidence for precise clinical management.Methods:A retrospective analysis was conducted on the clinicopathologic data of 1 445 patients with classical PTC treated at Xiangya Hospital of Central South University between August 2021 and June 2022.Patients were divided into groups based on the presence or absence of lymph node metastasis.Restricted cubic spline analysis explored the nonlinear relationship between BMI and lymph node metastasis risk.Univariate and multivariate Logistic regression analyses were applied to identify independent risk factors.Furthermore,sex-stratified analysis was performed among overweight patients(BMI≥24 kg/m2)to determine sex-specific risk factors for lymph node metastasis.Results:Among all patients,716(49.6%)had lymph node metastasis.Univariate analysis showed that BMI,sex,age,tumor diameter,multifocality,and extrathyroidal extension were significantly associated with cervical lymph node metastasis in PTC patients(all P<0.05).A nonlinear positive correlation was observed between BMI and lymph node metastasis risk,which was more pronounced in male patients.Additionally,BMI was positively correlated with triglyceride levels and negatively correlated with high-density lipoprotein cholesterol.Sex-stratified analysis revealed that in overweight male patients,younger age(OR=0.954),larger tumor diameter(OR=1.085),and multifocality(OR=2.776)were independent risk factors for LNM;in overweight female patients,younger age(OR=0.943)and larger tumor diameter(OR=1.074)were the main influencing factors.Conclusion:Overweight is closely associated with cervical lymph node metastasis in PTC,and the high-risk factors for LNM differ between male and female overweight patients.Young age,larger tumor size,and multifocality in overweight males,and young age and larger tumors in overweight females indicate a higher risk of metastasis.It is recommended that high-risk populations receive enhanced preoperative evaluation and individualized lymph node dissection strategies to achieve precise treatment and improved risk control.

Objective To investigate the effects of angelica polysaccharides on diabetic retinopathy(DR)and possi-ble mechanisms of action.Methods DR mouse models were established by injecting streptozotocin into 18 C57BL/6J mice.The mice were then divided randomly into an Angelica polysaccharide group(given 289 mg·kg-1 Angelica polysac-charides),a positive control group(given 217 mg·kg-1 calcium hydroxybenzenesulfonate),and a model group(given an equal volume of saline),6 mice in each group.6 normal mice were selected as a blank group(given an equal volume of sa-line).The drug was administered by gavage once a day for 28 consecutive days.ARPE-19 cells were exposed to a high-glu-cose(25 mmol·L-1)culture medium to establish DR cell models,which were divided into a negative control group(con-trol vector),a lncRNA MEG3 group(lncRNA MEG3 overexpression vector),and a model group(DR cells without vector transfection).Cells not treated with 25 mmol·L-1glucose were taken as a blank group.All cells were cultured for 24 h.Fasting blood glucose levels were detected in mice.Hematoxylin-Eosin(HE)staining was performed to study the his-topathological changes in the retinal tissue of mice.Quantitative polymerase chain reaction(qPCR)was performed to de-tect the expression level of lncMEG3 mRNAs in the retinal tissue of mice.CCK-8 assay was used to measure the activity of ARPE-19 cells.Enzyme linked immunosorbent assay(ELISA)was used to detect IL-1β and IL-6 levels in mouse retinal tis-sues and ARPE-19 cells.The Western blot analysis was made to detect the levels of cellular pyroptosis-related proteins(in-cluding Caspase-1,Cleaved-Caspase-1,GSDMD,GSDMD-N,NLRP3,IL-1 β,and IL-18)in mouse retinal tissues and ARPE-19 cells.Results Blood glucose levels in the mice of the model group were higher than those in the mice of the blank group and lower than those in the mice of Angelica polysaccharide and positive control groups(all P＜0.05).HE staining results showed that compared with those of the blank group,the mice in the mode group had significantly damaged retinal tissues and disordered and loose cell arrangement.The retinal tissue of the mice in Angelica polysaccharide and positive control groups was arranged more orderly and densely and suffered less damage,compared with that of the mice in the model group.The relative expression levels of lncMEG3 mRNAs in the retinal tissue of mice in the blank,model group,An-gelica polysaccharide,and positive control groups were 1.005±0 114,0.423±0.054,0.701±0.101 and 0.593±0.084,re-spectively.The retinal expression levels of lncMEG3 mRNAs in the mice of Angelica polysaccharide and positive control groups were significantly higher than those in the mice of the model group(both P＜0.01).CCK-8 results showed that the survival rates of ARPE-19 cells in the lncRNA MEG3 group at 48 and 72 h were higher than those in the negative control group(both P＜0.05).ELISA and Western blot results showed that the retinal expression levels of IL-1 β,IL-6 and cellular pyroptosis-related proteins in the mice of the model group were significantly higher than those in the mice of the blank group(all P＜0.01)and higher than those in the mice of Angelica polysaccharide group and positive control group(all P＜0.05).The expression levels of IL-1 β,IL-6,and cellular pyroptosis-related proteins in ARPE-19 cells of the model group were significantly higher than those in the blank group(all P＜0.01).The expression levels of IL-1 β,IL-6,and cellular py-roptosis-related proteins in ARPE-19 cells of the lncRNA MEG3 group were lower than those in the negative control group(all P＜0.05).Conclusion Angelica polysaccharides can improve DR by promoting lncRNA MEG3 expression and down-regulating IL-1 β,IL-6,Caspase-1,Cleaved-Caspase-1,GSDMD,GSDMD-N,NLRP3,IL-1β,and IL-18 protein ex-pression.

Objective To observe the application effect of bedside contrast-enhanced ultrasound combined with gas-water alternating injection method in severe patients with nasointestinal catheterization.Methods A total of 150 severe patients who were admitted to intensive care unit(ICU)and emergency intensive care unit(EICU)and required nasointestinal catheterization were collected.Patients were separated into the blind insertion method group(catheterization without other auxiliary equipment,n=50),the ultrasound method group(ultrasound-guided catheterization,n=53),and the combined method group(bedside contrast-enhanced ultrasound combined with gas-water alternating injection method,n=47)according to the wishes of patients or their families.The catheterization success rate,feeding standard-reaching rate within one week,catheterization time and incidence of adverse events were compared between three groups.Kappa test was used to compare the consistency of three methods with X-ray examination results after catheterization.Receiver operating characteristic(ROC)curve was used to evaluate the catheterization effect.Results The catheterization success rate and the feeding standard-reaching rate within one week were increased successively in the blind insertion method group,the ultrasound method group and the combined method group,and the catheterization time shortened in turn(P＜0.05).The consistency of blind insertion method,ultrasound method,combined method with X-ray examination was strong(Kappa=0.730),very strong(Kappa=0.835)and very strong(Kappa=0.911),respectively.Results of ROC curve showed that the areas under the ROC curve of the blind insertion method group,the ultrasound method group and the combined method group increased in turn,which were 0.838(95%CI:0.661-1.000),0.918(95%CI:0.763-1.000)and 0.988(95%CI:0.959-1.000),and the combined method group had the best catheterization effect.There were no significant differences in the incidence of adverse events such as hiccup,abdominal distension,diarrhea,aspiration and gastrointestinal bleeding between the three groups(P>0.05).Conclusion Bedside contrast-enhanced ultrasound combined with gas-water alternating injection method can improve the success rate of nasointestinal catheterization in severe patients,shorten the catheterization time and without increasing the incidence of adverse events.

Objective To explore the brain damage of SD rats under different time points of hypobaric hypoxia exposure.Methods A rat high-altitube cerebral edema(HACE)model was constructed by simulating an altitude of 6 000 m in a hypobaric hypoxia animal experimental chamber.Thirty-six SD male rats were randomly divided into the control group and the hypobaric hypoxia exposure 3,7 and 14 d groups,with 9 rats in each group.Except for the control group,the rats in each group were continuously exposed to hypobaric hypoxia for 3,7,and 14 d.At the end of the modeling period,serum was collected by blood sampling via the abdominal aorta,and brain tissue samples were taken.The wet-to-dry ratio(W/D)of brain tissue was calculated,and the levels of relevant oxidative enzymes in serum and brain tissue were measured.The expression levels of hypoxia-inducible factor-1α(HIF-1α)and aquaporin 4(AQP4)mRNAs in brain tissue were detected by real-time fluorescence quantitative polymerase chain reaction.Results The W/D of brain tissues in the control group and the group exposed to hypobaric hypoxia for 3,7 and 14 d were 4.46±0.12,4.98±0.16,5.07±0.18 and 4.95±0.07;the superoxide dismutase contents were(111.86±2.45),(90.73±1.48),(79.64±2.56)and(55.33±1.45)U·g-1;the glutathione contents were(126.91±5.18),(125.26±1.53),(56.20±2.17)and(122.73±1.78)μg·mL-1;the malondialdehyde contents were(230.94±2.00),(362.65±3.28),(407.34±3.47)and(237.50±1.59)nmol·g-1;the relative expression levels of HIF-1 α mRNA were 1.00±0,2.99±0.49,4.72±0.49 and 1.91±0.28;the relative expression levels of AQP4 mRNA were 1.00±0,2.62±0.34,8.38±0.84 and 5.27±0.42,respectively.Statistically significant differences were found between the above indexes in the 3,7 and 14 d of hypobaric hypoxia exposure group compared with the control group(P＜0.05,P＜0.01).Conclusion Different time of hypobaric hypoxia exposure can up-regulate the expression of AQPs proteins in HACE rats and cause the disruption of the blood-brain barrier,and the HACE model constructed in the hypobaric hypoxia chamber with 6 000 m intervention for 7 d was more stable.

Objective:To evaluate the efficacy of cyclophosphamide in patients with T-cell large granular lymphocytic leukemia (T-LGLL) and the maintenance of treatment-free remission (TFR) following drug discontinuation.Methods:Clinical data were collected from 37 patients with T-LGLL who received oral cyclophosphamide at the Regenerative Medicine Clinic of the Institute of Hematology and Blood Diseases Hospital between June 2019 and March 2024. Patient clinical characteristics, treatment efficacy, and long-term TFR were analyzed.Results:The median age of the 37 patients was 60 years (range: 37-86), and 22 (59.5%) were male. Anemia was observed in 30 patients (81.1%), and 28 (75.7%) met the diagnostic criteria for secondary pure red cell aplasia. Neutropenia occurred in 15 patients (40.5%), lymphocytosis in 11 (29.7%), and thrombocytopenia in three (8.1%). Sixteen patients (43.2%) had not received prior immunosuppressive therapy (treatment-naive group), while 21 patients (56.8%) were refractory to or had relapsed after immunosuppressive treatment (refractory/relapsed group). All patients met the treatment criteria and received oral cyclophosphamide at doses of 50-100 mg/day. Among the 36 evaluable patients, hematologic remission was achieved in 25 (69.4%), with a median time of 2.0 months (range: 0.7-7.0). There was no statistically significant difference in remission rates between the treatment-naive and refractory/relapsed groups (68.5% vs. 66.7%, P=0.589). Among the 25 patients who achieved hematologic remission, 24 discontinued cyclophosphamide. With a median follow-up of 39.0 months (range: 8.0-56.0), the median TFR duration was not reached. The estimated TFR rates were (90.87± 6.16) % at 12 months and (75.72±11.04) % at 36 months. No significant difference in TFR was observed between the treatment-naive and refractory/relapsed groups ( P=0.451) . Conclusion:Oral cyclophosphamide is effective in the treatment of T-LGLL, and patients may maintain long-term TFR following drug discontinuation.

Objective:To investigate the mechanisms of baicalin in treating septic acute liver injury through a combination of network pharmacology and animal experiments.Methods:Thirty male C57BL/6 mice (6 weeks old) were divided into five groups ( n=6): control group (normal saline), model group [lipopolysaccharide (LPS) 10 mg/kg, intraperitoneal injection], low-dose baicalin group (10 mg/kg), high-dose baicalin group (20 mg/kg), and baicalin-only group (20 mg/kg, without LPS). Baicalin was administered orally for 14 consecutive days prior to modeling. Mice were sacrificed 24 h after LPS injection. Alanine transaminase, aspartate transaminase liver tissue histopathology were measured; neutrophil infiltration was visualized using immunofluorescence; mRNA expression levels of interleukin (IL)-1β, IL-17, IL-6, and tumor necrosis factor (TNF)-α were detected by RT-qPCR; and the expression of Toll-like receptor 4 (TLR4) and phosphorylated nuclear factor (NF)-κB proteins were analyzed by Western blotting. Results:In the LPS model group, the ALT, AST, and histopathological injury score were (148.60±22.02) U/L, (81.58±11.59) U/L, and 8.50(7.75, 9.25), respectively. These indicators were significantly reduced in the high-dose baicalin group with (77.90±16.79) U/L, (49.92±14.89) U/L, and 1.00(1.00, 2.25) (all P<0.05). Compared with the LPS group, neutrophil infiltration in the liver of high-dose baicalin group was also significantly reduced [1.18%(0.98%, 1.22%) vs. 6.13%(5.41%, 8.69%), P<0.05]. RT-qPCR results showed that the relative mRNA expression levels of inflammatory cytokines IL-1β [(1.03±0.06) vs. (2.60±0.34)], IL-17 [(1.21±0.12) vs. (2.94 ± 0.39)], IL-6 [(1.37±0.26) vs. (2.73±0.18)], and TNF-α [(1.18±0.10) vs. (3.30±0.92)] were significantly decreased in the high-dose baicalin group compared with the LPS group (all P<0.05). Western blot analysis revealed that the relative protein expression levels of TLR4 [(1.25±0.13) vs. (1.73±0.06)] and phosphorylated NF-κB [(1.25±0.25) vs. (1.79±0.12)] were also significantly lower in the high-dose baicalin group (both P<0.05). Conclusion:Baicalin reduces liver injury in septic mice by downregula-ting the expression of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, and IL-17, potentially through the inhibition of the TLR4/NF-κB signaling pathway.

Objective To develop an enteral nutrition delivery device for nasal feeding patients to relieve gastrointestinal discomfort during enteral nutrition therapy and solve the problems in heating,quality preservation and constant temperature maintenance of the solution.Methods The enteral nutrition delivery device was mainly composed of a nasal feeding container,a fixing frame,a limiting trough,a heating wire,a temperature control unit and a speed control unit.The heating wire and temperature and speed control units were combined to facilitate the nasogastric solution in the container to be delivered to the patient's stomach and intestines at a constant temperature and a uniform speed.The device developed had its shell made of 304 waterproof stainless steel and the nasal feeding container made of 05-polypropylene.Results The device developed realized constant-temperature and uniform-speed delivery and auto cleaning,and could reduced effectively gastrointestinal adverse reactions during nasal feeding.Conclusion The device developed gains advantages in easy operation and autonomous feeding,and thus is worth promoting for nasal feeding patients clinically.[Chinese Medical Equipment Journal,2025,46(2):108-111]