Immune checkpoint inhibitors (ICIs) are widely used in the treatment of malignant tumors, and their related immune-related adverse events (irAEs) have attracted increasing attention. This study reports the diagnosis and treatment process of a case of immune cystitis in a patient with hepatobiliary tract malignant tumor after treatment with pembrolizumab. The patient was admitted to the hospital due to frequent urination, urgency of urination and dysuria for 1 month. Previous repeated anti-infection treatments were ineffective. Combined with medical history, laboratory tests, imaging findings, cystoscopy and pathological results, the patient was clinically diagnosed with ICIs-associated immune cystitis (Pembrolizumab) ultimately. The patient's symptoms significantly improved after treatment with glucocorticoids. This case reindicates that clinicians need to improve awareness of ICI-related urinary system irAEs. Early identification and timely intervention can significantly improve patient prognosis.

Background The escalating challenges of fine particulate matter (PM_2.5) pollution and global warming pose significant threats to human health. Previous studies have demonstrated that prenatal exposure to PM_2.5 and heat is associated with restricted fetal and offspring growth. However, the combined effects of co-exposure to PM_2.5 and heat remain poorly understood, and the underlying biological mechanisms have yet to be elucidated. Objective To investigate the independent and combined effects of maternal exposure to PM_2.5 and heat on offspring growth and thyroid hormone levels in rats. Methods Eight-week-old SPF Wistar rats were used. Females and males were mated at a 2:1 ratio. After confirming pregnancy, 36 pregnant rats were randomly assigned to four groups: combined PM_2.5 and heat exposure, PM_2.5 exposure, heat exposure, and control. PM_2.5 exposure was administered via intratracheal instillation (0.3 mL, 2 mg·mL⁻¹) daily. Heat exposure was conducted in a programmable constant temperature and humidity chamber maintained at (38.0±1.0)℃ for 1 h daily. Exposures were administered from gestational days (GD) 7 to 13. Offspring were raised for 42 postnatal days (PND), with body weight measured twice weekly at fixed time points. On PND42, blood was collected to separate plasma for the measurement of free triiodothyronine (FT₃) and free thyroxine (FT₄) levels using enzyme-linked immunosorbent assay. Linear mixed-effects models were used to examine the interactive effects of PM_2.5 and heat exposure on offspring body weight and weight gain. One-way analysis of variance was conducted to compare the differences in offspring body weight at each time point and thyroid hormone levels among groups. Correlations between thyroid hormone levels and weight changes were assessed using Spearman's rank correlation analysis. Results Both prenatal PM_2.5 exposure alone and its combination with heat exposure were associated with higher offspring body weight at specific postnatal time points. Offspring in the PM_2.5 exposure group had higher body weight than controls on PND7 and PND21, while those in the combined exposure group had higher body weight on PND7 and PND21-35 (P<0.05), and this effect was stronger in female offspring. The heat exposure group showed significantly lower birth weight than other groups (P<0.05), but this difference gradually disappeared during development. Plasma FT₃ levels were higher in the PM_2.5 exposure group than in the heat exposure and the control groups (P<0.05), but this difference was statistically significant only in female offspring. Plasma FT3 levels were positively correlated with body weight from PND7 to PND42, with correlation coefficients of 0.316–0.539 in all offspring (P<0.05) and 0.419–0.706 in females (P<0.05), while no significant correlation was observed in males. Conclusion Prenatal exposure to PM_2.5 alone or a combination of PM_2.5 and heat may affect postnatal body weight gain in offspring in a time-dependent manner, with more pronounced effects observed in female offspring. Heat exposure alone leads to reduced early-stage weight followed by a trend toward catch-up growth. The findings suggest a specific role of thyroid hormones in the effects of prenatal PM_2.5 and heat exposure on offspring growth, a relationship that warrants further investigation.

OBJECTIVE To investigate the effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease （NAFLD）. METHODS Thirty-two SD rats were randomly divided into normal group and modeling group. The modeling group was fed a high-fat diet to establish a NAFLD model. The successfully modeled rats were then randomly divided into model group， atorvastatin group[positive control， 2 mg/（kg·d）]， and Jingangteng capsules low- and high-dose groups [0.63 and 2.52 mg/（kg·d）]， with 6 rats in each group. The pathological changes of the liver were observed by hematoxylin-eosin staining and oil red O staining. Enzyme-linked immunosorbent assay was performed to determine the serum levels of triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine transaminase （ALT）， aspartate transaminase （AST）， tumour necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， IL-18. 16S rDNA amplicon sequencing and metabolomics techniques were applied to explore the effects of Jingangteng capsules on gut microbiota and metabolisms in NAFLD rats. Based on the E-mail：591146765@qq.com metabolomics results， Western blot analysis was performed to detect proteins related to the nuclear factor kappa-B （NF-κB）/NOD-like receptor family protein 3 （NLRP3） signaling pathway in the livers of NAFLD rats. RESULTS The experimental results showed that Jingangteng capsules could significantly reduce the serum levels of TG， TC， LDL-C， AST， ALT， TNF-α， IL-1β， IL-6， IL-18， while increased the level of HDL-C， and alleviated the hepatic cellular steatosis and inflammatory infiltration in NAFLD rats. They could regulate the gut microbiota disorders in NAFLD rats， significantly increased the relative abundance of Romboutsia and Oscillospira， and significantly decreased the relative abundance of Blautia （P＜0.05）. They also regulated metabolic disorders primarily by affecting secondary bile acid biosynthesis， fatty acid degradation， O-antigen nucleotide sugar biosynthesis， etc. Results of Western blot assay showed that they significantly reduced the phosphorylation levels of NF-κB p65 and NF-κB inhibitor α， and the protein expression levels of NLRP3， caspase-1 and ASC （P＜0.05 or P＜0.01）. CONCLUSIONS Jingangteng capsules could improve inflammation， lipid accumulation and liver injury in NAFLD rats， regulate the disorders of gut microbiota and metabolisms， and inhibit NF-κB/NLRP3 signaling pathway. Their therapeutic effects against NAFLD are mediated through the inhibition of the NF-κB/NLRP3 signaling pathway.

Objective: To compare the changes in the concentration of relevant growth factors released from platelet-rich plasma (PRP) stored at -80℃ by cryopreservation and at 4℃ by refrigerated lyophilization over 2 years, aiming to provide a theoretical basis for prolonging PRP storage duration. Methods: PRP (n=15) was separated using a blood cell separator and stored under -80℃ cryopreservation (F-PRP group) and 4℃ refrigerated freeze-drying conditions (FD-PRP group). The contents of growth factors (PDGF-AA, PDGF-BB, EGF, TGF-β1, and VEGF) in both groups were measured by ELISA at 1, 3, 6, 9, 12 and 24 months. Results: PDGF-AA and VEGF maintained good stability in both groups for up to 24 months. PDGF-BB and TGF-β1 showed high stability in the first 12 months but their stability decreased gradually from 12th to 24th months. EGF demonstrated good stability in the first 6 months, and its stability gradually decreased from the 9th to 24th months. Comparing the F-PRP and FD-PRP groups, the concentrations of the five growth factors in the FD-PRP group were either not statistically different or higher than those in the F-PRP group at all time points. Specifically, the concentrations of EGF were significantly higher in the FD-PRP group at all time points. Conclusion: Both -80℃ freezing and 4℃ freeze-drying enable long-term preservation of PRP. Freeze-drying imposes less stringent storage requirements and facilitates growth factor compared to frozen storage.

Objective: To investigate the potential protective effect of Shexiang Tongxin dropping pills (STDP) on ischemia-reperfusion injury and its underlying mechanisms in improving endothelial cell function in coronary microvascular disease (CMVD). Methods: A rat model of myocardial ischemia-reperfusion injury with CMVD was established using ligation and reperfusion of the left anterior descending artery. The effect of STDP (21.6 mg/kg) on cardiac function was evaluated using echocardiography, hematoxylin-eosin staining, and Evans blue staining. The effects of STDP on the microvascular endothelial barrier were assessed based on nitric oxide production, endothelial nitric oxide synthase expression, structural variety of tight junctions (TJs), and the expression of zonula occludens-1 (ZO-1), claudin-5, occludin, and vascular endothelial (VE)-cadherin proteins. The mechanisms of STDP (50 and 100 ng/mL) were evaluated by examining the expression of sphingosine 1-phosphate receptor 2 (S1PR2), Ras Homolog family member A (RhoA), and Rho-associated coiled-coil-containing protein kinase (ROCK) proteins and the distribution of ZO-1, VE-cadherin, and F-actin proteins in an oxygen and glucose deprivation/reoxygenation model. Results: The administration of STDP on CMVD rat model significantly improved cardiac and microvascular endothelial cell barrier functions (all P < .05). STDP enhanced the structural integrity of coronary microvascular positioning and distribution by clarifying and completing TJs and increasing the expression of ZO-1, occludin, claudin-5, and VE-cadherin in vivo (all P < .05). The S1PR2/RhoA/ROCK pathway was inhibited by STDP in vitro, leading to the regulation of endothelial cell TJs, adhesion junctions, and cytoskeletal morphology. Conclusion STDP showed protective effects on cardiac impairment and microvascular endothelial barrier injury in CMVD model rats induced by myocardial ischemia-reperfusion injury through the modulation of the S1PR2/RhoA/ROCK pathway.

This study aims to explore the mechanism of lung and gut protection by Tanreqing Capsules on the mice infected with influenza virus based on "the lung-gut axis". A total of 110 C57BL/6J mice were randomized into control group, model group, oseltamivir group, and low-and high-dose Tanreqing Capsules groups. Ten mice in each group underwent body weight protection experiments, and the remaining 12 mice underwent experiments for mechanism exploration. Mice were infected with influenza virus A/Puerto Rico/08/1934(PR8) via nasal inhalation for the modeling. The lung tissue was collected on day 3 after gavage, and the lung tissue, colon tissue, and feces were collected on day 7 after gavage for subsequent testing. The results showed that Tanreqing Capsules alleviated the body weight reduction and increased the survival rate caused by PR8 infection. Compared with model group, Tanreqing Capsules can alleviate the lung injury by reducing the lung index, alleviating inflammation and edema in the lung tissue, down-regulating viral gene expression at the late stage of infection, reducing the percentage of neutrophils, and increasing the percentage of T cells. Tanreqing Capsules relieved the gut injury by restoring the colon length, increasing intestinal lumen mucin secretion, alleviating intestinal inflammation, and reducing goblet cell destruction. The gut microbiota analysis showed that Tanreqing Capsules increased species diversity compared with model group. At the phylum level, Tanreqing Capsules significantly increased the abundance of Firmicutes and Actinobacteria, while reducing the abundance of Bacteroidota and Proteobacteria to maintain gut microbiota balance. At the genus level, Tanreqing Capsules significantly increased the abundance of unclassified＿f＿Lachnospiraceae while reducing the abundance of Bacteroides, Eubacterium, and Phocaeicola to maintain gut microbiota balance. In conclusion, Tanreqing Capsules can alleviate mouse lung and gut injury caused by influenza virus infection and restore the balance of gut microbiota. Treating influenza from the lung and gut can provide new ideas for clinical practice.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Lung/metabolism* ; Mice, Inbred C57BL ; Capsules ; Orthomyxoviridae Infections/virology* ; Gastrointestinal Microbiome/drug effects* ; Male ; Humans ; Female ; Influenza A virus/physiology* ; Influenza, Human/virology*

This study aimed to characterize and identify the non-volatile components in aqueous and ethanolic extracts of the stems and leaves of Rhododendron tomentosum by using sensitive and efficient ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UHPLC-Q-TOF-MS) combined with a self-built information database. By comparing with reference compounds, analyzing fragment ion information, searching relevant literature, and using a self-built information database, 118 compounds were identified from the aqueous and ethanolic extracts of R. tomentosum, including 35 flavonoid glycosides, 15 phenolic glycosides, 12 flavonoids, 7 phenolic acids, 7 phenylethanol glycosides, 6 tannins, 6 phospholipids, 5 coumarins, 5 monoterpene glycosides, 6 triterpenes, 3 fatty acids, and 11 other types of compounds. Among them, 102 compounds were reported in R. tomentosum for the first time, and 36 compounds were identified by comparing them with reference compounds. The chemical components in the ethanolic and aqueous extracts of R. tomentosum leaves and stems showed slight differences, with 84 common chemical components accounting for 71.2% of the total 118 compounds. This study systematically characterized and identified the non-volatile chemical components in the ethanolic and aqueous extracts of R. tomentosum for the first time. The findings provide a reference for active ingredient research, quality control, and product development of R. tomentosum.
Rhododendron/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/chemistry* ; Mass Spectrometry/methods* ; Plant Leaves/chemistry*

Objective: To investigate the expressions of GPR15 and FOXP3 in colorectal carcinoma(CRC) tissues and their clinical prognostic values. Methods : A total of 132 patients with CRC underwent radical surgery were collected. The control group selected the normal mucosal tissues more than 5 cm away from the edge of the cancer focus. Immunohistochemistry(Envision two-step method) was used to detect the expression levels of GPR15 and FOXP3 in CRC and adjacent tissues, and analyze their relationships with clinicopathological factors of colorectal cancer. Kaplan-Meier method was used to draw the survival curve to analyze the correlation between the expressions of GPR15 and FOXP3 and the survival prognosis of patients with CRC. The factors influencing prognosis of patients with colorectal cancer were analyzed by Cox regression. Results : The immunohistochemistry showed that the expression levels of GPR15 and FOXP3 in CRC were significantly higher than those in normal colorectal mucosal tissues(P<0.05). The expression of GPR15 in CRC tissues was correlated with location, nerve invasion and TNM stage; FOXP3 expression was correlated with sex(P<0.05).Both expressions were not significantly correlated with the clinicopathologic features of age, tumor size, differentiation degree, tissue type, depth of invasion, tumor budding, vascular invasion and lymph node metastasis. Correlation analysis showed that there was no significant correlation between GPR15 and FOXP3 expression(Kappa=-0.019,P>0.05). The survival prognosis of GPR15 positive group was significantly worse than that of negative group(log-rank: χ2=4.3,P=0.039);while the survival prognosis of FOXP3 positive group was significantly better than that of negative group(log-rank: χ2=7.3,P=0.007).Age ≤55 years, positive GPR15 and negative FOXP3 were independent risk factors for poor prognosis in patients with CRC(P<0.05). Conclusion The expression levels of GPR15 and FOXP3 in CRC are significantly higher than those in paracancer tissues, GPR15 and FOXP3 are expected to become new tumor markers for early screening, accurate treatment and prognosis assessment of CRC.

Objective:To establish a quality control method for Aiye standard decoction.Methods:The ultra performance liquid chromatogrphy (UPLC) column Waters ACQUITY HSS T3 C18 (2.1 mm×150 mm,1.8 μm) was used to gradient elution by acetonitrile and 0.1% formic acid in water. 16 batches of Aiye standard decoction fingerprints were established by UPLC and the common peaks were determined in the fingerprints. The contents of 12 components were determined. The 16 batches of Aiye standard decoction were analyzed by similarity calculation, hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) for analysis of differential components of Artemisiae Argyi Folium from different origins.Results:A total of 13 common peaks were marked in the fingerprints of 16 batches of Aiye standard decoction, 12 of which were identified by comparison with reference substance, including chlorogenic acid, sochlorogenic acid A, neochlorogenic acid, cryptochlorogenic acid, caffeic acid,1,3-O-Dicaffeoylquinic acid, schaftoside, isochlorogenic acid B,1,5-O-Dicaffeoylquinic acid, isochlorogenic acid C, jaceosidin and eupatilin. Similarity evaluation, PCA and HCA all classified the 16 batches of Aiye standard decoction into 2 categories. Orthogonal partial least squares discriminant analysis screened 5 differential biomarkers from 13 common peaks. The content determination results showed that the phenolic compounds and flavonoids in samples from Hubei were significantly higher than that in samples from other areas.Conclusion:This method can effectively analyze the differences in the quality of Aiye standard decoction from different origins, and provide reference for the formulation of quality standards for Aiye standard decoction and related preparations.

Objective:To establish a method for the determination of triterpenes and nucleosides in Poria based on HPLC; To accurately determine the various bioactive components in Poria.Methods:Similarity evaluation, clustering analysis and principal component analysis were used to analyze the similarities and differences of different medicinal parts of Poria, and the key chromatographic peaks that could reflect the characteristics were found.Results:The Poricoic acid A and dehydroeburiconic acid could be used as the identification basis for Poriae Cutis and White Poria; at the same time, Polyporenic acid C, dehydropachymic acid and dehydrotrametenolic acid could be used to evaluate Rubra Poria and Poriae Cutis; uridine, guanosine and adenosine may be essential ingredients for evaluating the quality of White Poria, Poriae Cutis and Rubra Poria. In different medicinal parts of Poria, the triterpenes were showed significant differences; by contrary, there were little differences among the same medicinal parts.Conclusion:This study reveals the quality differences between different medicinal parts of Poria, which can provide a scientific basis for the rational application and pharmacodynamic standardization of Poria.