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

Identification of disease-specific cell subtypes (DSCSs) has profound implications for understanding disease mechanisms, preoperative diagnosis, and precision therapy. However, achieving unified annotation of DSCSs in heterogeneous single-cell datasets remains a challenge. In this study, we developed the gPRINT algorithm (generalized approach for cell subtype identification with single cell's voicePRINT). Inspired by the principles of speech recognition in noisy environments, gPRINT transforms gene position and gene expression information into voiceprints based on ordered and clustered gene expression phenomena, obtaining unique "gene print" patterns for each cell. Then, we integrated neural networks to mitigate the impact of background noise on cell identity label mapping. We demonstrated the reproducibility of gPRINT across different donors, single-cell sequencing platforms, and disease subtypes, and its utility for automatic cell subtype annotation across datasets. Moreover, gPRINT achieved higher annotation accuracy of 98.37% when externally validated based on the same tissue, surpassing other algorithms. Furthermore, this approach has been applied to fibrosis-associated diseases in multiple tissues throughout the body, as well as to the annotation of fibroblast subtypes in a single tissue, tendon, where fibrosis is prevalent. We successfully achieved automatic prediction of tendinopathy-specific cell subtypes, key targets, and related drugs. In summary, gPRINT provides an automated and unified approach for identifying DSCSs across datasets, facilitating the elucidation of specific cell subtypes under different disease states and providing a powerful tool for exploring therapeutic targets in diseases.
Humans ; Algorithms ; Single-Cell Analysis ; Databases, Genetic ; Molecular Sequence Annotation

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*

Objective To study in vitro anti-HIV activity of an extract of herb medicines,SanJiangDan,and the possible mechanism.Methods The main active ingredient of SanJiangDan was extracted by distillation.Three subtypes of HIV pseudovirus (B subtype,C subtype,CRF01_AE subtype) were used to evaluate the anti-HIV activity of SanJiangDan extract in vitro.The possible mechanism was evaluated through analyzing the effects of SanJiangDan extract on the expression of surface receptors and cytokines by T cells.The cytotoxicity of SanJiangDan extract was detected by using four different sources of cell lines including epithelial cells Caco-2 cells,TZM cells,Huh7 cells derived from liver cells and lymphocyte Jurkat-T cells.Results SanJiangDan extract effectively inhibited the infection of HIV pseudoviruses at concentrations of 1.6 mg/ml and 0.16 mg/ml.The inhibition rates were 30.9％,36.6％ and 65.0％ for B subtype,C subtype and CRF01_AE subtype respectively at the concentration of 0.16 mg/ml.As the concentration increased to 1.6 mg/ml,the inhibition rates increased to 96.4％ (B subtype),97.4％ (C subtype) and 99.5％ (CRF01_AE subtypes),but no toxicity to host cells was detected.Moreover,SanJiangDan extract inhibited the expression of HIV surface receptors including CD4,CXCR4 and CCR5 on TZM-bl cells,but enhanced IL-2 production.Conclusion SanJiangDan extract could inhibit HIV pseudovirus infection without causing cytotoxicity to host cells in vitro.The possible mechanism might be associated with the reduced expression of CD4,CXCR4 and CCR5 and enhanced secretion of IL-2 as well.

OBJECTIVETo investigate the transcriptional regulation of pacemaker channel I(f) mediated by vasoactive peptide endothelin-1 (ET-1) in neonatal rat ventricular myocytes and its mechanism.

METHODSNeonatal rat ventricular myocytes were enzymatically isolated. I(f) current was recorded using the whole-cell patch-clamp technique. The expression of hyperpolarization-activated cyclic nucleotide-gated channel (HCN) isoforms HCN2 and HCN4 were measured by quantitative RT-PCR.

RESULTSET-1 increased the expression of HCN2 and HCN4 mRNA in a dose- and time-dependent manner. These effects were blocked by specific ETA receptor antagonist BQ-123 but not the ETB receptor antagonist BQ-788. The effects of ET-1 on HCN2 and HCN4 mRNA expression were not affected by the p38 mitogen-activated protein kinase (MAPK) inhibitor (SB-203580).

CONCLUSIONThese findings indicate that ET-1 stimulates the expression of pacemaker channel I(f) in cardiomyocytes via ETA receptor through a p38 MAPK-independent signaling pathway, which might be linked to the intrinsic arrhythmogenic potential of ET-1.

Animals ; Animals, Newborn ; Cyclic Nucleotide-Gated Cation Channels ; drug effects ; Endothelin-1 ; metabolism ; Imidazoles ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; Oligopeptides ; pharmacology ; Patch-Clamp Techniques ; Piperidines ; pharmacology ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

Sexual transmission of HIV-1 has recently become the predominant route over the world.As an effective HIV vaccine has been elusive so far,microbicides,which are usually applied at topical mucosal sites to prevent vaginal or rectal mucosa from HIV-1 infection,have been paid ever increasing attentions.The failure of previous clinical trials have demonstrated that it is critical to ensure the safety and efficacy of microbicides before they move forward into clinical trials.In this review,we have summarized recent progress in preclinical models evaluating the safety and efficacy of microbicides,including cell culture in vitro,mucosal explant and animal models in vivo which are used at different phases during the preclinical development of microbicides,and have elaborated the characteristics of these models and their advantages.Finally,we emphasize the urgent need to establish effective evaluation systems at molecular levels for different models,which may serve as good surrogates to predict the inflammation and HIV infection in future clinical testing.

Objective To explore the relationship between invasive growth and expression of ER、PTTG、bFGF in human prolactinomas.Methods Fourty-six prolactinomas specimens were collected and divided into invasive group and non-invasive group.Expressions of ER、PTTG、bFGF were examined by immunocytochemical method.Total RNA was extracted from specimens and ER-mRNA and PTTG-mRNA were detected by RT-PCR.Results IA of protein expression of ER、bFGF、PTTG in invasive prolactinomas was seperately 5385.1?1348.6、9874.2?2143.7、7938.5?1436.5;The optical density ratio of ER-mRNA and PTTG-mRNA was 0.71 and 0.83,difference between invasive group and non-vasive group was significant.Conclusions ER、PTTG、bFGF in human prolactinomas closely correlated with invasiveness of prolactinomas,and may play important function in prolactinomas.

Objective To investigate the relationships between the expressions of estrogen receptor (ER), proliferating cell nuclear antigen (PCNA) in prolactinomas and tumor invasiveness. Methods The expressions of ER and PCNA proteins were examined by immunocytochemical method, including 24 invasive prolactinomas (invasive group) and 32 non-invasive prolactinomas (non-invasive group). The expression of ER-mRNA was detected by reverse transcription-polymerase chain raction (RT-PCR) technique. Results There were 23 cases of positive protein expression of ER in invasive group, the integral optical density (IOD) was 4935.12?1246.56, significantly lower than non-invasive group ( P