BACKGROUND: T cell dysfunction, which includes exhaustion, anergy, and senescence, is a distinct T cell differentiation state that occurs after antigen exposure. Although T cell dysfunction has been a cornerstone of cancer immunotherapy, its potential in transplant research, while not yet as extensively explored, is attracting growing interest. Interferon regulatory factor 4 (IRF4) has been shown to play a pivotal role in inducing T cell dysfunction. METHODS: A novel ultra-low-dose combination of Trametinib and Rapamycin, targeting IRF4 inhibition, was employed to investigate T cell proliferation, apoptosis, cytokine secretion, expression of T-cell dysfunction-associated molecules, effects of mitogen-activated protein kinase (MAPK) and mammalian target of rapamycin (mTOR) signaling pathways, and allograft survival in both in vitro and BALB/c to C57BL/6 mouse cardiac transplantation models. RESULTS: In vitro , blockade of IRF4 in T cells effectively inhibited T cell proliferation, increased apoptosis, and significantly upregulated the expression of programmed cell death protein 1 (PD-1), Helios, CD160, and cytotoxic T lymphocyte-associated antigen (CTLA-4), markers of T cell dysfunction. Furthermore, it suppressed the secretion of pro-inflammatory cytokines interferon (IFN)-γ and interleukin (IL)-17. Combining ultra-low-dose Trametinib (0.1 mg·kg -1 ·day -1 ) and Rapamycin (0.1 mg·kg -1 ·day -1 ) demonstrably extended graft survival, with 4 out of 5 mice exceeding 100 days post-transplantation. Moreover, analysis of grafts at day 7 confirmed sustained IFN regulatory factor 4 (IRF4) inhibition, enhanced PD-1 expression, and suppressed IFN-γ secretion, reinforcing the in vivo efficacy of this IRF4-targeting approach. The combination of Trametinib and Rapamycin synergistically inhibited the MAPK and mTOR signaling network, leading to a more pronounced suppression of IRF4 expression. CONCLUSIONS Targeting IRF4, a key regulator of T cell dysfunction, presents a promising avenue for inducing transplant immune tolerance. In this study, we demonstrate that a novel ultra-low-dose combination of Trametinib and Rapamycin synergistically suppresses the MAPK and mTOR signaling network, leading to profound IRF4 inhibition, promoting allograft acceptance, and offering a potential new therapeutic strategy for improved transplant outcomes. However, further research is necessary to elucidate the underlying pharmacological mechanisms and facilitate translation to clinical practice.
Animals ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Interferon Regulatory Factors/metabolism* ; Heart Transplantation/methods* ; T-Lymphocytes/immunology* ; Sirolimus/therapeutic use* ; Pyridones/therapeutic use* ; Graft Survival/drug effects* ; Pyrimidinones/therapeutic use* ; Cell Proliferation/drug effects* ; Apoptosis/drug effects* ; Male ; Signal Transduction/drug effects*

OBJECTIVES: The integrated endoplasmic reticulum stress inhibitor (ISRIB) is a selective inhibitor of the protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway within endoplasmic reticulum stress (ERS) and can improve spatial and working memory in aged mice. Although ERS and oxidative stress are tightly interconnected, it remains unclear whether ISRIB alleviates cognitive impairment by restoring the balance between ERS and oxidative stress. This study aims to investigate the effects and mechanisms of ISRIB on lipopolysaccharide (LPS)-induced cognitive impairment in mice. METHODS: Eight-week-old male ICR mice were randomly divided into 3 groups: Normal saline (NS) group, LPS group, and ISRIB+LPS group. NS and LPS groups received daily intraperitoneal injections of normal saline for 7 days; on day 7, LPS group mice received intraperitoneal LPS (0.83 mg/kg) to establish a cognitive impairment model. ISRIB+LPS group received ISRIB (0.25 mg/kg) intraperitoneally for 7 days, with LPS injected 30 minutes after ISRIB on day 7. Cognitive ability was evaluated by the novel place recognition test (NPRT). Real-time fluorogenic quantitative PCR (RT-qPCR) was used to detect changes in nitric oxide synthase (NOS), superoxide dismutase-1 (SOD-1), and catalase (CAT) gene expression in the hippocampus and prefrontal cortex. Oxidative stress markers malondialdehyde (MDA), glutathione (GSH), and oxidized glutathione (GSSG), were measured in hippocampal and prefrontal cortex tissues. RESULTS: Compared with the NS group, mice in LPS group showed a significant reduction in novel place recognition ratio, upregulation of hippocampal NOS-1 and NOS-2 mRNA, downregulation of SOD-1 and CAT mRNA, increased MDA and GSSG, decreased GSH, and reduced GSH/GSSG ratio (all P<0.05). Compared with the LPS group, mice in ISRIB+LPS group exhibited significantly improved novel place recognition, downregulated NOS-1 and NOS-2 mRNA, upregulated SOD-1 and CAT mRNA, decreased MDA and GSSG, increased GSH, and an elevated GSH/GSSG ratio in the hippocampus (all P<0.05). No significant changes were observed in the prefrontal cortex. CONCLUSIONS ISRIB improves LPS-induced cognitive impairment in mice by restoring the oxidative/antioxidant balance in the hippocampus.
Animals ; Lipopolysaccharides ; Male ; Mice, Inbred ICR ; Cognitive Dysfunction/drug therapy* ; Mice ; Oxidative Stress/drug effects* ; Endoplasmic Reticulum Stress/drug effects* ; Hippocampus/drug effects* ; Nitric Oxide Synthase Type II/genetics* ; Guanidines/pharmacology* ; eIF-2 Kinase/antagonists & inhibitors* ; Signal Transduction/drug effects* ; Superoxide Dismutase/metabolism*

Objective:To observe the effects of kidney-tonifying and mind-calming acupuncture therapy on sleep,mood,sex hormone levels,and traditional Chinese medicine(TCM)symptoms in patients with perimenopausal insomnia(PMI). Methods:A total of 90 patients with PMI were randomly divided into a treatment group and a control group,with 45 cases in each group.Patients in the treatment group were treated with acupuncture at Shenshu(BL23),Taixi(KI3),Baihui(GV20),and Anmian(Extra).The control group was treated with sham acupuncture.Both groups were treated 3 times a week for 4 weeks.Pittsburgh sleep quality index(PSQI)and insomnia severity index(ISI)were used to evaluate the sleep quality of the subjects before treatment,after treatment,and 1 month after treatment(follow-up).Beck depression inventory(BDI)and Beck anxiety inventory(BAI)were used to evaluate the depression and anxiety of the subjects before treatment,after treatment,and at 1-month follow-up.The TCM symptom scale was used to evaluate the TCM symptoms of the subjects before treatment,after treatment,and 1 month after treatment.Serum levels of estradiol(E2),follicle-stimulating hormone(FSH),and luteinizing hormone(LH)were measured before and after treatment. Results:During the study,2 cases dropped out of the treatment group,and no cases dropped out of the control group.The PSQI scores of the treatment group were significantly lower after treatment and at 1-month follow-up compared with those before treatment(P<0.05),and the difference was statistically significant compared with that of the control group(P<0.05).In the control group,the PSQI score was significantly lower after treatment compared with before treatment(P<0.05),and the difference was not statistically significant at 1-month follow-up compared with before treatment(P>0.05).Compared with the pre-treatment,the ISI,BDI,BAI,and TCM symptom scale scores of the treatment group were lower after treatment and at 1-month follow-up(P<0.05),and the differences with the control group at the same time point were statistically significant(P<0.05).The differences in ISI,BDI,BAI,and TCM symptom scale scores of the control group before treatment,after treatment,and at 1-month follow-up were not statistically significant(P>0.05).After treatment,the serum E2 level in the treatment group was significantly higher than that before treatment(P<0.05),and the difference with the control group was statistically significant(P<0.05).The difference in the serum E2 level before and after treatment in the control group was not statistically significant(P>0.05).The differences in the serum FSH and LH levels between before and after treatment were not statistically significant in either group of subjects(P>0.05). Conclusion:Kidney-tonifying and mind-calming acupuncture therapy can improve sleep quality,relieve anxiety and depression,delay the decrease of serum E2 level,and improve related TCM symptoms in patients with PMI.

Background::Homoharringtonine (HHT) is an effective anti-inflammatory, anti-viral, and anti-tumor protein synthesis inhibitor that has been applied clinically. Here, we explored the therapeutic effects of HHT in a mouse heart transplant model.Methods::Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver, kidney, and hematology. A mouse heart transplantation model was constructed, and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan–Meier analysis, immunostaining, and bulk RNA sequencing analysis. The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells (Tregs) differentiation. Results::HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo. Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days ( P <0.001) without non-immune toxicity. The allografts had long-term survival after continuous HHT treatment for 28 days. HHT significantly reduced lymphocyte infiltration in the graft, and interferon-γ-secreting CD4 + and CD8 + T cells in the spleen ( P <0.01). HHT significantly increased the number of peripheral Tregs (about 20%, P <0.001) and serum interleukin (IL)-10 levels. HHT downregulated the expression of T cell receptor (TCR) signaling pathway-related genes ( CD4, H2-Eb1, TRAT1, and CD74) and upregulated the expression of IL-10 and transforming growth factor (TGF) -β pathway-related genes and Treg signature genes ( CTLA4, Foxp3, CD74, and ICOS). HHT increased CD4 + Foxp3 + cells and Foxp3 expression ex vivo, and it enhanced the inhibitory function of inducible Tregs. Conclusions::HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels, thereby promoting mouse heart allograft acceptance. These findings may have therapeutic implications for organ transplant recipients, particularly those with viral infections and malignancies, which require a more suitable anti-rejection medication.

Objective: To evaluate the effectiveness of varicella vaccine in varicella outbreaks and to analyze the influencing factors, and to provide a reference for making the targeted prevention and controlling measures. Methods: A total of 3 888 students with no history of varicella were selected from 2 schools with varicella outbreak in Guangdong Province in 2021, a retrospective cohort study was conducted by using questionnaire survey, rate ratio ( RR ) and vaccine effectiveness ( VE ) values were calculated and Logistic regression was uses to analyze the factors influencing the protective effect of varicella. Results: There were 138 confirmed cases of varicella among the participants. There was no significant sex difference in the vaccination rate( χ 2=1.36, P =0.51), but there was significant difference in the vaccinattion rate of different age groups( χ 2=555.82, P <0.01). The overall protective effect of VarV was 66.94%(95% CI =56.17%-77.71%), and the protective effect of 2 doses of vaccine( VE = 90.02% , 95% CI =83.13%-96.90%) was higher than that of 1 dose( VE =49.40%, 95% CI =32.36%-66.44%)( χ 2=24.93, P < 0.01 ). The high fever rates in the vaccinated and unvaccinated groups were 7.69% and 25.81%, with significant difference( χ 2= 6.29 , P <0.05). The rates of moderate and severe skin lesions of vaccinated and unvaccinated groups was 20.00% and 50.00%, respectively, and the difference was statistically significant( χ 2=11.32, P <0.01). The protective effects of varicella vaccine against high fever and moderate to severe rash were 70.19%(95% CI =42.11%-98.27%) and 60.00%(95% CI =38.15%-81.85%). Stratified analysis showed that there were significant differences in different years of vaccination( χ 2=37.87, P <0.05), while there were no significant differences in age of vaccination and vaccine manufacturer ( P >0.05). Conclusion Varicella vaccination can prevent chickenpox infection and reduce the severity of the disease. However, the efficacy of varicella vaccine was affected by vaccination years. It is recommended to improve the vaccination coverage of varicella vaccine to prevent the outbreak of the epidemic.

Objective:To observe the effects of acupuncture and moxibustion on phosphatase and PTEN-induced putative kinase 1(PINK1)/Parkin signaling pathway in the midbrain substantia nigra of Thy1-α synuclein(αSyn)transgenic model mice with Parkinson disease(PD). Methods:Twenty-four Thy1-αSyn transgenic mice were randomly divided into a model group,an acupuncture group,an acupuncture + moxibustion group,and a Western medicine group.Six wild-type mice in the same litter were used as the wild-type group.In the acupuncture group,Baihui(GV20)and Yanglingquan(GB34)were selected for acupuncture.In the acupuncture + moxibustion group,Guanyuan(CV4)was added on the basis of the acupuncture group.The Western medicine group was given rapamycin intraperitoneal injection at a dose of 10 mg/(kg·bw).The wild-type group and the model group were fixed without intervention.The overall rod performance(ORP)score of mice was observed in each group.The immunohistochemical method was used to detect the tyrosine hydroxylase(TH)positive neurons in the substantia nigra of mice in each group.The αSyn was detected by the immunofluorescence chemical method.The expression levels of αSyn,microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ,autophagy protein sequestosome-1/protein 62(SQSTM-1/p62),PINK1,Parkin,and ubiquitin-specific protease 30(USP30)proteins were detected by Western blotting assay.The expression levels of LC3B,p62,PINK1,Parkin,and USP30 mRNAs were detected by fluorescence quantitative polymerase chain reaction. Results:Compared with the wild-type group,the ORP score,the p62,PINK1,and Parkin protein expression levels decreased significantly(P<0.01),the PINK1 mRNA expression level decreased(P<0.05),while the protein and mRNA expression levels of USP30 increased(P<0.05)in the model group.Compared with the model group,the ORP score in the acupuncture group and the acupuncture + moxibustion group increased(P<0.05);the expression level of LC3-Ⅱ/LC3-Ⅰ protein in the acupuncture + moxibustion group and the Western medicine group increased(P<0.05);the protein expression levels of p62,PINK1,and Parkin increased(P<0.05),while the USP30 protein expression level decreased significantly(P<0.01)in the acupuncture group,the acupuncture + moxibustion group,and the Western medicine group;the Parkin mRNA expression level in the acupuncture group and the acupuncture + moxibustion group increased(P<0.05);the USP30 mRNA expression level in the acupuncture + moxibustion group decreased(P<0.05). Conclusion:Acupuncture and moxibustion regulate the related molecule expression levels of PINK1/Parkin signaling pathway in the Thy1-αSyn transgenic PD model mice and promote the autophagy degradation of αSyn to exert the protective effect of dopaminergic neurons.

OBJECTIVES: To investigate the relationship between the fatty acid-binding protein 4 (FABP4) and colorectal cancer (CRC). METHODS: Using an enzyme-linked immunosorbent assay (ELISA), we measured the expression of FABP4 in plasma of 50 patients who underwent surgery for CRC from October 2017 to May 2018 and 50 healthy controls. The content of the visceral fat area (VFA) as seen with abdominal computed tomography (CT) scanning was measured by ImageJ software. The expression levels of FABP4, E-cadherin, and Snail proteins in CRC and adjacent tissues were determined by immunohistochemistry. RESULTS: The mean concentration of plasma FABP4 of CRC patients was higher than that of the control group (22.46 vs. 9.82 ng/mL; CONCLUSIONS High LPA and VFA were risk factors for increased plasma FABP4 in CRC patients. FABP4 protein was highly expressed in CRC tissues and associated with TNM stage, differentiation, and lymph node metastasis of CRC. The level of FABP4 in CRC tissue was correlated with E-cadherin and Snail expression, suggesting that FABP4 may promote CRC progression related to epithelial-mesenchymal transition (EMT).

BACKGROUND: Microglia are resident immune cells of the central nervous system that normally perform sensing, housekeeping, and defense functions. In the context of neurodegenerative diseases, the dysfunction of microglia leads to or aggravates neuronal injury. OBJECTIVE: To investigate the mechanism of microglia-mediated neuronal injury in neurodegenerative diseases. METHODS: The first author searched for relevant articles published from January 2001 to January 2020 in PubMed, CNKI, Wanfang database, and VIP database with the key words of “microglia; neurodegenerative diseases; neuronal injury”. RESULTS AND CONCLUSION: In neurodegenerative diseases, microglia perform excessive sensing due to toxic substances during normal function, leading to increasing activation of microglia, accompanied with hyperfunction of housekeeping and intense neuroinflammation causing neuronal impairment. The dysregulation can also be manifested as dysfunction of sensing and housekeeping due to specific gene mutations, which bring about accumulation of toxic substances, aggravating the dysregulation of defense function and inducing apoptosis or necrosis of neurons as a result. Further exploration on the mechanism of microglia-mediated neuronal injury in neurodegenerative diseases may provide several targets for the treatment of neurodegenerative disease.

Objective:To analyze the epidemiological characteristics of the cases firstly reported as "asymptomatic infection of COVID-19" in Guangdong province.Methods:The follow-up observation method was used to continuously track and observe the cases firstly reported as "asymptomatic patients with COVID-19" in Guangdong province from January 14 to March 31, 2020. The epidemiological data of the cases were collected to analyze their epidemiological characteristics, outcome and influencing factors.Results:From January 14 to March 31, 2020, a total of 325 cases were firstly reported as "asymptomatic infections of COVID-19" in Guangdong province. The epidemic curve of asymptomatic infection cases was similar to that of confirmed cases, and it had two peaks. The first peak was from January 27 to February 5, and the second peak was from March 17 to March 26. Of the 325 cases, 184 (56.6%) were subsequently converted to confirmed cases. These cases were defined as incubation period asymptomatic infection cases. The age median of the cases was 40 years, and 93.5% (172/184) of the cases showed symptoms within 3 days after the first positive nucleic acid tests were conducted, and 141 (43.4%) of the 325 cases remained asymptomatic status until they were cured and discharged. They were inapparent infection cases, accounting for 8.6% (141/1 642) of those diagnosed with COVID-19 in Guangdong province during the same period. The age median of inapparent infection cases was 27 years. The median of the interval between the first positive nucleic acid test and discharge was 14 days. Up to 90.8% (138/141) of the inapparent infection cases were discharged for centralized medical observation within 28 days. The longest interval between the first positive nucleic acid test and the last positive nucleic acid test was 73 days. The positive rate of nucleic acid test was 0.3% in close contacts of inapparent infection cases and 2.2% in close contacts of incubation period asymptomatic infection cases. There were significant differences in age distribution and source of infection between incubation period asymptomatic infection cases and inapparent infection cases ( P<0.05). Old age was the risk factor for the conversion of firstly reported asymptomatic infection cases to confirmed cases. Compared with the 0-19-year-old group, The patients aged 40-59 years and 60 years and above were more likely to become confirmed cases. The OR (95 %CI) values were 2.730 (1.380-5.402) and 5.302 (2.199-12.783), and P values were 0.004 and 0.000, respectively. People being infected in China were more likely to become confirmed cases ( OR=7.121, P=0.000). Conclusions:There were asymptomatic infection cases among patients diagnosed with COVID-19. The infectiousness of incubation period asymptomatic infection cases might be stronger than that of inapparent infection cases. The proportion of younger cases among asymptomatic infection cases was higher than that of the confirmed cases. Old age and domestic infection were the risk factors for the conversion of asymptomatic infection cases to confirmed cases, to which more attention should be paid. Further serological investigations are needed to provide a basis for the development of COVID-19 prevention and control strategies.

Objective To observe the clinical effect on sleep improvement about acupoint application in treating the chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.Methods A total of 68 patients with chronic insomnia with Liver-stagnation and Spleen-deficiency Syndrome were randomly and blindly divided into treatment group and control group by registration order. In the treatment process, 1 was eliminated and 33 were completed in the treatment group, 2 were eliminated and 32 were completed in the control group. The treatment group underwentAn Mian Tietherapy half an hour before bed and the control group underwent placebo therapy in the same way of treatment group. Two groups were treated for 40 days and followed-up visit six months. The change of Pittsburgh Sleep Quality (PSQI), Index Insomnia Severity Index (ISI) and TCM Syndromes Scale were detected.Results The clinical total effective rate of treatment group was 72.7% and the control group was 9.4%, and the difference was statistically significant (χ2=46.977,P<0.01). The PSQI scores after treatment (7.55 ± 1.52vs. 13.90 ± 2.44,t=148.165), and follow up 1 month (8.97 ± 2.51vs. 13.17 ± 2.79,t=37.926) in the treatment group were significantly lower than those in the control group (P<0.01). The ISI scores after treatment (7.03 ± 3.37vs. 20.89 ± 4.40,t=73.75), and follow up 1 month (9.81 ± 3.16vs. 19.41 ± 3.66,t=40.79) in the observation group were significantly lower than those in the control group (P<0.01). The TCM Syndromes Scale scores after treatment (2.05 ± 1.09vs. 6.98 ± 1.23,t=17.116), and follow up 1 month (4.06 ± 1.59vs. 6.83 ± 0.91,t=68.055) and follow up 6 month (5.12 ± 1.84vs. 7.19 ± 1.07,t=27.716) in the observation group were significantly lower than those in the control group (P<0.01).Conclusions Acupoint application could obviously change the sleep quality and Chinese medicine symptom in chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.