Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

Objective To analyze the composition and function of residual cells in pre-transplantation human thymic slices by single-cell transcriptomics sequencing(scRNA-seq),and established a quality assessment method for thymic slices based on the expression levels of molecular markers in the culture supernatant.Methods The discarded thymus from 18 patients with congenital heart disease undergoing surgical treatment in Department of Cardiothoracic Surgery of Children's Hospital Affiliated to Chongqing Medical University from May 2023 to January 2024 were collected and prepared into thymic slices.After the slices were cultured in vitro for 14 d,scRNA-seq was employed to identify the residual cell types,and gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis was performed to analyze the biological function of the residual cells.Then based on the literature concerning thymic slice culture,the molecular markers indicating thymocyte function were screened out.ELISA was applied to detect the changes in protein levels of molecular markers in the supernatant.Receiver operating characteristic(ROC)curve was plotted and assess the value of the molecular markers in the supernatant in evaluating the quality of thymic slices with area under the curve(AUC).Then,the qualified and unqualified thymic slices determined by our obtained molecular markers were transplanted subcutaneously into male nude mice(6～8 weeks old,weighing 14～17 g),respectively,and the male nude mice without transplantation of the thymic slices served as control group.Flow cytometry and histologic analysis were utilized to observe the immune reconstitution after transplantation.Results ① scRNA-seq identified 11 cell types in thymic slices,dominated with epithelial cells,fibroblasts,and T cells.GO and KEGG enrichment analysis showed that epithelial cells were involved in enrichment entries related to chemotaxis,epithelial cell development,cell matrix adhesion and tight junction;fibroblasts were involved in enrichment entries related to extracellular matrix,epithelial cell proliferation,negative regulation of cell migration,and regulation of actin cytoskeleton;T cells were mainly related to T cell differentiation,regulation of T cell activation,T cell apoptosis,and T cell receptor signaling.② Molecular markers,CCL19,CCL21,CXCL12,CXCL16,IL16 and SELL were identified to indicate thymocyte function.Compared with the levels of the first day,the protein secretions of CCL19,CCL21,CXCL12 and CXCL16 were significantly increased during in vitro culture(P<0.05),while the protein secretions of IL16 and L-selectin(protein form of SELL)were significantly decreased(P<0.05).The combined predictor Pre1 from subset of cytokines(IL16 and L-selectin)had the highest value in the quality assessment of thymic slices after 1 d of culture(AUC=0.883),and the combined predictor Pre2 from subset of cytokines(CCL19,CCL21,CXCL12 and CXCL16)had the highest value in the quality assessment after 14 d of culture(AUC=0.948).③ Transplantation in nude mice indicated that the qualified thymic slices could develop to thymus structure in vivo,and effectively increase the proportion of T cells in peripheral blood(P<0.01),while the unqualified thymic slices could not obtain the reconstitution of T cell development.Conclusion The main residual component cells in thymic slices are epithelial cells,fibroblasts and T cells.IL16 and L-selectin can be used as potential indicators to determine the quality of donor thymic samples.CCL19,CCL21,CXCL12 and CXCL16 can effectively evaluate the quality of thymic slices before transplantation.

OBJECTIVE This study investigated the clinical implications of endothelial cell-specific molecule 1(ESM-1)in obstructive sleep apnea(OSA)patients,with particular focus on its dynamic correlation with adhesion molecules,aiming to elucidate the regulatory role of ESM-1 in OSA-associated vascular endothelial impairment.METHODS This cross-sectional study enrolled participants undergoing polysomnography(PSG)at the Sleep Medicine Center of Beijing Anzhen Hospital,Capital Medical University between March 2017 and January 2018.Based on the inclusion criteria,161 participants were ultimately included and divided into OSA group(n=118)and control group(n=43).Demographic data and polysomnography parameters were collected.We used a powerful high-throughput Multiplex Immunobead Assay technology to simultaneously test plasm cytokines levels of ESM-1,inter-cellular adhesion molecule 1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1).Circulating C-reactive protein(CRP)and homocysteine(Hcy)were detected by routine blood chemistry panel.RESULTS Circulating ESM-1 levels were significantly elevated in patients with OSA compared with healthy controls[819.73(612.36-1393.47)pg/ml]vs.[286.17(114.48-513.81)pg/ml,P<0.001].After adjusting for confounding factors,we found that circulating ESM-1 levels were an independent risk factor for OSA(odds ratio=2.162,95%CI=1.522-3.072,P<0.001)and circulating ESM-1 levels were positively associated with ICAM-1 and VCAM-1 levels(β=1.977,95%CI=1.429-2.734,P<0.001).CONCLUSION Circulating ESM-1 levels were significantly increased in patients with OSA,which is closely related with adhesion molecules levels.ESM-1 may be a surrogate endothelial dysfunction marker and an independent risk factor for OSA.

Objective:To develop rat models of oral mucosa ulcer using distinct experimental methodologies,fulfilling research requirements and adhering to the ethical standards for animal care.Methods:96 SD rats were randomly allocated into groups.The rats in control group(n=8)were regularly fed without other treatment.Those in chemical cauterization groups were treated by 20％,40％,60％of glacial acetic acid(GAA)on oral mucosa(n=8);in mechanical damage groups by 30 000 r/min high speed drill induced trauma of 10,20 and 30 mm2 respectively(n=8);in ionizing radiation groups were treated with 10,12,15,20 and 30 Gy on the mucosa respectively(n=8).After the ulcer was appeared,the morphology of the mucosa were observed,the mucosal tissue lesions were examined by HE,Masson and immunofluorescence staining,the expression of TNF-α and IL-1β were detected by qPCR and ELISA respectively,and the body conditions such as diet and body weight of the rats were observed,the pain,dis-tress and discomfor of the rats were evaluated by MORTON&Griffits Guidelines.Results:40％and 60％GAA,20 mm2 and 30 mm2 friction damage and ionizing radiation of 12 Gy or greater may induce oral mucosa ulcer with a diseas coruse of 6-7 d in SD rats.TNF-α and IL-1β mRNA expression in the damaged tissue,the related protein expression in blood serum of the rats were in-creased.MORTON&Griffits Guidelines analysis showed 40％GAA,20 mm2 friction damage and 12 Gy ionizing radiation induced the lowest scores of pain,distress and discomfort of the rats with compatible oral mocosa ulcere induced by the relevat treatment.Conclusion:40％GAA,20 mm2 of friction damage and 12 Gy of ionizing radiation can reliably establish oral mucosa ulcer models and minimize adverse effects on SD rats,and accord with ethical standards of 3R for laboratory animal.

This study reports the diagnosis and treatment of six cases of post-transplant lymphoproliferative disorder (PTLD) in liver transplant recipients, confirmed at the Affiliated Hospital of Qingdao University between August 2017 and May 2023. The report includes details on anti-rejection therapy, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infections, imaging findings, histopathological results, treatment courses, and prognoses. By summarizing the clinical experience in the diagnosis and management of PTLD following liver transplantation, this study aims to provide valuable insights and references for the clinical diagnosis and treatment of this condition.

Objective:To study and establish the UPLC fingerprint and multi-index content determination methods of Hedyotidis chrysotricahae Herba; To provide a reference for the quality control of Hedyotidis chrysotricahae Herba.Methods:The chromatographic column was ACQUITY UPLC HSS T3 (100 mm×2.1 mm,1.8 μm). The mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution; the detection wavelength was 254 nm; the flow rate was 0.30 ml/min and column temperature was 35 ℃. The method could determine content and fingerprint of rutin, Kaempferol-3-O-rutinoside, Narcissoside, Neochlorogenic aci, Chlorogenic Acid, Cryptochlorogenic acid and have quality analysis to 17 batches of Hedyotidis chrysotricahae Herba based on the variance of fingerprint, similarity evaluation, clustering analysis along with principal component analysis (PCA) at the same time.Results:The common pattern of UPLC specific chromatogram of Hedyotidis chrysotricahae Herba was established. The 11 common peaks were marked out, among which 7 peaks were identified. 17 batches Hedyotidis chrysotricahae Herba could be divided into 4 categories according to different origins. Quality content of six indicators of Hedyotidis chrysotricahae Herba was in slight difference between different origins, among which the content quality of Hedyotidis chrysotricahae Herba from Duyun in Guizhou Province was the highest.Conclusion:The established UPLC fingerprint and content determination method of 6 indicators from the study can be used for the quality control of Hedyotidis chrysotricahae Herba, which can also provide a theoretical basis for the standard improvement of Hedyotidis chrysotricahae Herba.

Objective:To develop rat models of oral mucosa ulcer using distinct experimental methodologies,fulfilling research requirements and adhering to the ethical standards for animal care.Methods:96 SD rats were randomly allocated into groups.The rats in control group(n=8)were regularly fed without other treatment.Those in chemical cauterization groups were treated by 20％,40％,60％of glacial acetic acid(GAA)on oral mucosa(n=8);in mechanical damage groups by 30 000 r/min high speed drill induced trauma of 10,20 and 30 mm2 respectively(n=8);in ionizing radiation groups were treated with 10,12,15,20 and 30 Gy on the mucosa respectively(n=8).After the ulcer was appeared,the morphology of the mucosa were observed,the mucosal tissue lesions were examined by HE,Masson and immunofluorescence staining,the expression of TNF-α and IL-1β were detected by qPCR and ELISA respectively,and the body conditions such as diet and body weight of the rats were observed,the pain,dis-tress and discomfor of the rats were evaluated by MORTON&Griffits Guidelines.Results:40％and 60％GAA,20 mm2 and 30 mm2 friction damage and ionizing radiation of 12 Gy or greater may induce oral mucosa ulcer with a diseas coruse of 6-7 d in SD rats.TNF-α and IL-1β mRNA expression in the damaged tissue,the related protein expression in blood serum of the rats were in-creased.MORTON&Griffits Guidelines analysis showed 40％GAA,20 mm2 friction damage and 12 Gy ionizing radiation induced the lowest scores of pain,distress and discomfort of the rats with compatible oral mocosa ulcere induced by the relevat treatment.Conclusion:40％GAA,20 mm2 of friction damage and 12 Gy of ionizing radiation can reliably establish oral mucosa ulcer models and minimize adverse effects on SD rats,and accord with ethical standards of 3R for laboratory animal.

This study reports the diagnosis and treatment of six cases of post-transplant lymphoproliferative disorder (PTLD) in liver transplant recipients, confirmed at the Affiliated Hospital of Qingdao University between August 2017 and May 2023. The report includes details on anti-rejection therapy, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infections, imaging findings, histopathological results, treatment courses, and prognoses. By summarizing the clinical experience in the diagnosis and management of PTLD following liver transplantation, this study aims to provide valuable insights and references for the clinical diagnosis and treatment of this condition.

This article reports a patient with hepatic coma who underwent artificial liver support therapy and liver transplantation successfully， and the patient recovered well in the later stage after active treatment. This article also discusses the timing of liver transplantation.

Objective To investigate the status of human papillomavirus(HPV)infection in women in Li-aocheng area,and to analyze the relationship between HPV genotypes and thin-prep cytologic test(TCT),so as to provide reference for the diagnosis,treatment and vaccination of cervical cancer in this area.Methods The relevant data of 4 968 women who underwent physical examination in the hospital from Januar-y 2020 to April 2023 were retrospectively analyzed.Cervical exfoliated cells were collected for TCT and HPV genotyping,and the distribution of HPV infection and genotype in women of different ages was analyzed.x2 test was used to compare the differences between groups,and multivariate Logistic regression was used to ana-lyze the influencing factors of different HPV types on cervical lesions.Results The overall HPV infection rate was 21.05％,and there was a significant difference in the overall infection rate among different age groups(x2=42.695,P＜0.001).The HPV infection rate of＞49-59 years old group was the highest(26.85％),followed by＞59 years old group.Among the different types of HPV infection,single infection accounted for the highest proportion(15.12％),followed by double infection(4.07％).The top five high-risk genotypes of HPV infection rate were HPV52,16,58,53,66,and HPV16,52,58 were the main high-risk genotypes in HPV single infection.TCT distribution of 1 046 HPV-positive cases:54.97％had no intraepithelial lesion or malig-nant lesion,29.64％atypical squamous cells of undetermined significance,2.58％atypical squamous cells could not exclude high-grade squamous intraepithelial lesion,11.47％low-grade squamous intraepithelial le-sion,and 1.34％high-grade squamous intraepithelial lesion.The infection rates of HPV16,58 and 66 were re-lated to the severity of cervical cytology(x2 trend=23.765,7.807,16.122,P＜0.05).Multivariate Logistic regression analysis showed that HPV16,18,31,33,35,39,51,52,53,56,58,59,66 genotypes were associated with cervical lesions.Conclusion The risk of HPV infection is high in women in this area,especially in mid-dle-aged and elderly women.High-risk HPV types 52,16 and 58 are the most common infection types in this area.HPV16 is closely related to the degree of cervical lesions.