This study aims to investigate the effect of Chaijin Jieyu Anshen Formula on the neuroinflammation of rats with insomnia complicated with depression through the regulation of triggering receptor expressed on myeloid cells 2(TREM2)/complement protein C1q signaling pathway. Rats were randomly divided into a normal group, a model group, a positive drug group, as well as a high, medium, and low-dose groups of Chaijin Jieyu Anshen Formula, with 10 rats in each group. Except for the normal group, the other groups were injected with p-chlorophenylalanine and exposed to chronic unpredictable mild stress to establish the rat model of insomnia complicated with depression. The sucrose preference experiment, open field experiment, and water maze test were performed to evaluate the depression in rats. Enzyme-linked immunosorbent assay was employed to detect serum 5-hydroxytryptamine(5-HT), dopamine(DA), and norepinephrine(NE) levels. Hematoxylin and eosin staining and Nissl staining were used to observe the damage in nucleus accumbens neurons. Western blot and immunofluorescence were performed to detect TREM2, C1q, postsynaptic density 95(PSD-95), and synaptophysin 1(SYN1) expressions in rat nucleus accumbens, respectively. Golgi-Cox staining was utilized to observe the synaptic spine density of nucleus accumbens neurons. The results show that, compared with the model group, Chaijin Jieyu Anshen Formula can significantly increase the sucrose preference as well as the distance and number of voluntary activities, shorten the immobility time in forced swimming test and the successful incubation period of positioning navigation, and prolong the stay time of space exploration in the target quadrant test. The serum 5-HT, DA, and NE contents in the model group are significantly lower than those in the normal group, with the above contents significantly increased after the intervention of Chaijin Jieyu Anshen Formula. In addition, Chaijin Jieyu Anshen Formula can alleviate pathological damages such as swelling and loose arrangement of tissue cells in the nucleus accumbens, while increasing the Nissl body numbers. Chaijin Jieyu Anshen Formula can improve synaptic damage in the nucleus accumbens and increase the synaptic spine density. Compared to the normal group, the expression of C1q protein was significantly higher in the model group, while the expression of TREM2 protein was significantly lower. Compared to the model group, the intervention with Chaijin Jieyu Anshen Formula significantly downregulated the expression of C1q protein and significantly upregulated the expression of TREM2. Compared with the model group, the PSD-95 and SYN1 fluorescence intensity is significantly increased in the groups receiving different doses of Chaijin Jieyu Anshen Formula. In summary, Chaijin Jieyu Anshen Formula can reduce the C1q protein expression, relieve the TREM2 inhibition, and promote the synapse-related proteins PSD-95 and SNY1 expression. Chaijin Jieyu Anshen Formula improves synaptic injury of the nucleus accumbens neurons, thereby treating insomnia complicated with depression.
Animals ; Male ; Rats ; Nucleus Accumbens/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Depression/complications* ; Membrane Glycoproteins/genetics* ; Rats, Sprague-Dawley ; Sleep Initiation and Maintenance Disorders/complications* ; Neurons/metabolism* ; Receptors, Immunologic/genetics* ; Signal Transduction/drug effects* ; Synapses/metabolism*

ObjectiveTo analyze the reported cases of infectious diseases across different tiers of public medical and healthcare institutions in Minhang District, Shanghai from 2013 to 2023, to investigate the status and changes in reported infectious diseases in this district from a temporal, etiological, and demographic perspectives, so as to provide a scientific basis for the construction of a hierarchica early-warning surveillance system under the grading diagnosis and treatment model in medical institutions, as well as for optimizing sentinel surveillance at facilities of different levels. MethodsA retrospective analysis was performed using surveillance data from the China Disease Prevention and Control Information System in Minhang District from 2013 to 2023. Reported infectious diseases were categorized into three categories based on transmission routes: respiratory infectious diseases, intestinal infectious diseases, and sexually transmitted and blood borne infectious diseases. According to the implementation phase of the grading diagnosis and treatment policy, the research time was divided into four time periods: 2013‒2016, 2017‒2019, 2020‒2022, and 2023. The distribution and temporal changes of reported cases of infectious diseases were compared across community health service centers (CHCs), secondary hospitals, tertiary grade-A hospitals and tertiary grade-B hospitals. Chi-square test was used for univariate analysis of differences in the number of reported cases. Quantitative data with normal distribution were analyzed using parametric tests, otherwise, Kruskal⁃Wallis H tests were used. ResultsThe proportions of total reported cases of infectious diseases in medical institutions at all levels in Minhang District, Shanghai from 2013 to 2023 were 10.66% in CHCs, 9.10% in secondary hospitals, 64.95% in tertiary grade-B hospitals, and 15.29% in tertiary grade-A hospitals, with an overall decline and then rebound trend in the reported cases. After the implementation of grading diagnosis and treatment policy, the number of reported cases in CHCs and secondary hospitals showed a trend of first decreasing and then increasing, while that in tertiary grade-B hospitals showed a steady decreasing trend and that in tertiary grade-A hospitals showed an increasing trend. In terms of the research periods divided above, a total of 10 392 cases were reported in 2013‒2016 (70.34% from tertiary grade-B hospitals and 12.59% from CHCs), including 2 922 cases of respiratory infectious diseases, 1 241 cases of intestinal infectious diseases, and 6 229 cases of sexually transmitted and blood-borne infectious diseases. Between 2017 and 2019, a total of 6 967 cases were reported (73.49% from tertiary grade-B hospitals and 11.84% from tertiary grade-A hospitals), including 2 983 cases of respiratory infectious diseases, 279 cases of intestinal infectious diseases, and 3 705 cases of sexually transmitted and blood-borne infectious diseases. Between 2020 and 2022, a total of 4 599 cases were reported (69.92% from tertiary grade-B hospitals and 24.57% from tertiary grade-A hospitals), including 1 627 cases of respiratory infectious diseases, 123 cases of intestinal infectious diseases, and 2 849 cases of sexually transmitted and blood-borne infectious diseases. In 2023, a total of 4 648 cases were reported (35.20% from tertiary grade-B hospitals and 27.50% from tertiary grade-A hospitals), including 3 165 cases of respiratory infectious diseases, 69 cases of intestinal infectious diseases, and 1 414 cases of sexually transmitted and blood-borne infectious diseases. The proportion of reported cases from tertiary grade-B hospitals was the highest in all the four research periods, but exhibited an obvious decrease in 2023. The differences in the reported cases of infectious diseases with different transmission routes among medical institutions at all levels were statistically significant (χ²=3 225.628, P<0.05). The differences in the mean age of patients among medical institutions at all levels were statistically significant (H=1 325.927, P<0.05). ConclusionThere are significant differences in the number of reported cases of infectious disease in the medical institutions at different levels. Tertiary grade-B hospitals have historically dominated the number of reported cases, but its share has declined recently. Whereas, CHCs and tertiary grade-A hospitals have played an increasingly important role in the surveillance and early warning of respiratory and intestinal infectious diseases. Therefore, it is recommended to leverage the strengths of grading diagnosis and treatment to establish targeted sentinel sites and deploy specialized teams tailored to the epidemiological characteristics of specific disease categories.

The innate immune sensor NLRP3 inflammasome overactivation is involved in the pathogenesis of ulcerative colitis. PGAM5 is a mitochondrial phosphatase involved in NLRP3 inflammasome activation in macrophages. However, the role of PGAM5 in ulcerative colitis and the mechanisms underlying PGAM5 regulating NLRP3 activity remain unknown. Here, we show that PGAM5 deficiency ameliorates dextran sodium sulfate (DSS)-induced colitis in mice via suppressing NLRP3 inflammasome activation. By combining APEX2-based proximity labeling focused on PGAM5 with quantitative proteomics, we identify NEK7 as the new binding partner of PGAM5 to promote NLRP3 inflammasome assembly and activation in a PGAM5 phosphatase activity-independent manner upon inflammasome induction. Interfering with PGAM5-NEK7 interaction by punicalagin inhibits the activation of the NLRP3 inflammasome in macrophages and ameliorates DSS-induced colitis in mice. Altogether, our data demonstrate the PGAM5-NEK7 interaction in macrophages for NLRP3 inflammasome activation and further provide a promising therapeutic strategy for ulcerative colitis by blocking the PGAM5-NEK7 interaction.

Objective:To explore the impact of the stimulator of interferon genes(STING)-interferon regulatory factor 3(IRF3)pathway on the senescence of rapid pacing HL-1 myocytes.Methods:HL-1 cells were divided into five groups: the control group(HL-1 cells without any treatment), pacing group(HL-1 cells paced for 48 hours), STING siRNA group(HL-1 cells paced for 48 hours and transfected with STING siRNA), NC siRNA group(HL-1 cells paced for 48 hours and transfected with NC siRNA), and H151 inhibitor group(HL-1 cells paced for 48 hours with the addition of 1 μmol/L STING inhibitor H151). Mitochondrial membrane potential was assessed in control and pacing group cells, and mitochondrial MitoTracker and TFAM co-localization staining was performed on these cells.Cellular senescence was evaluated using β-galactosidase staining in each group, and the positive rate of cellular senescence was observed and calculated.Western blotting was employed to detect the expression levels of STING, IRF3, P-IRF3, P16, P21, and P53 proteins in all groups.Immunofluorescence was utilized to examine the expression distribution of STING and P21 across the various groups.ELISA was performed to measure the concentrations of interleukin(IL)-1β, IL-6, and IL-8 in the cell supernatants from each group as part of the senescence-associated secretory phenotype(SASP).Results:Compared with the control group, the ratio of mitochondrial JC-1 multimer to monomer was significantly decreased in the pacing group( t=16.42, P<0.05), the co-localization of mitochondrial MitoTracker and TFAM in the cells was significantly weakened, the proportion of cells with positive cellular senescence-associated β-galactosidase staining significantly increased in the pacing group, the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 proteins were significantly elevated in the pacing group, and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants were markedly increased.Compared with the pacing group, the proportion of cells with positive cellular senescence-associated β-galactosidase staining decreased in the STING siRNA group and H151 inhibitor group ( F= 18.13, P<0.05), the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 were reduced in the STING siRNA group and H151 inhibitor group ( F=16.84, 26.56, 74.70, 31.80, 31.23, all P<0.05), and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants decreased( F=197.80、1 339.00、1 308.00, all P<0.001). Conclusions:Rapid pacing of HL-1 cells can promote mtDNA release into the cytoplasm, activate the STING-IRF3 pathway, accelerate cellular senescence, and enhance the secretion of SASP.Inhibiting the expression of STING can delay the senescence induced by the rapid pacing of HL-1 cells and reduce SASP secretion.

The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C₁₈ column and a conventional C₁₈ column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H₂O₂ measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H₂O₂ clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field.

Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Objective:To investigate the effects of warming needle combined with pelvic floor muscle massage on clinical symptoms,quality of life(QOL),and sex hormone levels in patients with benign prostatic hyperplasia(BPH).Methods:A total of 140 patients diagnosed with BPH were randomly divided into a control group,a warming needle group,a massage group,and a combined treatment group,with 35 patients in each group.The control group was given oral finasteride tablets for treatment.In addition to oral finasteride tablets,the warming needle group was given warming needle treatment,the massage group was given pelvic floor muscle massage,and the combined treatment group was given both warming needle and pelvic floor muscle massage.After two treatment courses,the total effective rate was compared among the four groups,and changes in such clinical indicators as international prostate symptom score(I-PSS),QOL score,total prostate volume(TPV),testosterone(T),estradiol(E2),luteinizing hormone(LH),and prostate-specific antigen(PSA)were observed.Results:The differences in the total effective rate among the four groups indicated statistical significance(P<0.05),and the combined treatment group showed a significantly higher effective rate than the other three groups(P<0.05).After treatment,the I-PSS,QOL score,TPV,and serum levels of T,E2,LH,and PSA in all four groups improved compared to those before treatment(P<0.05),and the extent of improvement in the combined treatment group was significantly greater than that in the other three groups(P<0.05).Conclusion:Oral finasteride tablets,warming needle,and pelvic floor muscle massage can all improve the I-PSS,QOL score,TPV,and serum levels of T,E2,LH,and PSA in patients with BPH,and the combination of the three therapies demonstrates the most significant effect.

Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Pulmonary electrical impedance tomography (EIT), a noninvasive, continuous, dynamic, and radiation-free bedside imaging technique for monitoring pulmonary ventilation, is now widely utilized in the diagnosis and management of critically ill patients. Beyond ventilation monitoring, hypertonic saline contrast-enhanced EIT for bedside pulmonary perfusion assessment has recently garnered significant attention. This article describes the application of hypertonic saline contrast-enhanced EIT to evaluate pulmonary perfusion in two patients following pulmonary endarterectomy, providing a reference for its perioperative application in such patients.