ObjectiveTo observe the effects of Yifei Tongluo prescription on the NOD-like receptor protein 3 （NLRP3）/Caspase-1/gasdermin D （GSDMD） pathway and epithelial-mesenchymal transition （EMT） in rats with pulmonary fibrosis. MethodsTracheal instillation of bleomycin was conducted to establish a rat model of pulmonary fibrosis. Thirty Sprague-Dawley （SD） rats were randomly divided into a blank group， a model group， a prednisone acetate group （1.17 mg·kg^-1）， and low- and high-dose Yifei Tongluo prescription groups （10.62 and 21.24 g·kg^-1， respectively）. Administration started on the 7th day after modeling， once a day for 28 consecutive days. The lung coefficient of each group was calculated. The pathological changes of lung tissues in each group were observed by hematoxylin-eosin （HE） staining and Masson staining. The expression of α-smooth muscle actin （α-SMA） and vimentin in rat lung tissues was detected by immunohistochemistry. The expression of NLRP3 inflammasome， E-cadherin （E-cad）， and typeⅠ collagen （ColⅠ） in lung tissues was detected by immunofluorescence. The content of hydroxyproline （HYP）， tumor necrosis factor （TNF）-α， interleukin （IL）-18， and IL-1β in rat serum was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression levels of NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， IL-1β， and transforming growth factor （TGF）-β₁ in rat lung tissues were determined by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of NLRP3， GSDMD， ASC， and Caspase-1 in rat lung tissues were determined by Western blot. ResultsCompared with the blank group， the model group exhibited a significantly increased lung coefficient （P<0.01） and significantly increased range of pulmonary interstitial inflammation and collagen deposition. In addition， the levels of α-SMA， Vimentin， E-cad， and ColⅠ in lung tissues were significantly increased （P<0.01）. The levels of fibrosis- and inflammation-related factors HYP， TNF-α， IL-18， and IL-1β in serum were significantly upregulated （P<0.01）. The levels of factors related to the activation of NLRP3 inflammasome in lung tissues， including NLRP3， GSDMD， ASC， Caspase-1， IL-1β， and TGF-β₁， were significantly upregulated （P<0.01）. Compared with the model group， the Yifei Tongluo prescription groups showed improved lung coefficients. Additionally， the extent of lung inflammation and collagen deposition was significantly reduced. The expression of α-SMA， Vimentin， E-cad， and ColⅠ in lung tissue was significantly decreased （P<0.01）. The levels of HYP， TNF-α， IL-18， and IL-1β in serum were significantly reduced （P<0.01）. The expression levels of NLRP3， GSDMD， ASC， Caspase-1， IL-1β， and TGF-β₁ in lung tissue were also significantly decreased （P<0.01）. ConclusionYifei Tongluo prescription can regulate the NLRP3/Caspase-1/GSDMD pathway， down-regulate release of pro-inflammatory and pro-fibrotic cytokines， alleviate NLRP3 inflammasome-mediated pyroptosis and EMT， and thereby improve pulmonary fibrosis in rats.

Objective: To establish a real-time quality control scheme based on the median (MD-IC) of internal control cycle threshold value in negative samples (NEG-IC-CT), so as to monitor anomalies such as progressive drift in nucleic acid testing system not covered by conventional internal quality control (IQC) in blood center nucleic acid laboratories, and to verify its feasibility. Methods: The internal control CT values of 54 426 negative samples were retrospectively collected. These samples were from four reagent batches of the two new and old equipment sets during the operation of the Wantai nucleic acid testing system in our blood center. The daily median of NEG-IC-CT values was used as the research indicator. Control limits were calculated using median absolute deviation (MAD) to construct the Median-MAD quality control chart. The monitoring performance of this scheme for the operation status of the testing system was simultaneously evaluated. Results: Statistical analysis showed significant differences in NEG-IC-CT value distribution between the new and old equipment sets, as well as between the two different reagent batches of the old equipment (P<0.000 1). The NEG-IC-CT value performance of the two different reagent batches of the new equipment was no significant difference in distribution (P>0.05). This scheme identified three typies of distinct anomalies. The out-of-control events observed with the old equipment in both the O1 and O2 reagent batches suggested potential performance decay due to equipment aging. The unreported change of reagent batch in time of Phase B with new equipment caused a stepwise drift on the quality control chart. In the later stage of Phase A with the new equipment, an alert was triggered, indicating potential quality risks associated with practices such as the mixed use of the remaining reagents and extremely long operator working hours. Conclusion: The realtime quality control scheme based on NEG-IC-CT value established in this study has been preliminarily validated for its monitoring effectiveness in nucleic acid testing in our blood center. This scheme performed well in detecting differences among testing systems and reagent batches, serving as an effective supplement to routine internal quality control. It can provide an intuitive and effective evaluation method for monitoring the performance of the nucleic acid testing process at blood center.

Background Pyrethroid pesticides (PYRs) can cross the placental barrier to cause intrauterine fetal exposure, which may lead to developmental immunotoxicity (DIT). However, the specific effect of maternal PYR exposure during pregnancy on the cellular immune function of 1-year-old children remains unclear. Objective To explore the effect of PYRs exposure throughout the entire pregnancy on peripheral blood lymphocytes in 1-year-old children and potential sensitive window period of PYRs exposure. Methods A birth cohort was established by enrolling pregnant women in their first trimester and following them and their infants until one year of age. Ultra-high performance liquid chromatography-tandem mass spectrometry was used to detect the levels of PYRs metabolites, including 3-phenoxybenzoic acid (3PBA), 4-fluoro-3-phenoxybenzoic acid (4F3PBA), and cis-3-(2,2-dichlorovinyl)-2,2- dimethylcyclopropane carboxylic acid (cis-DBCA), in the urine of pregnant women during the first trimester (gestational weeks 6-12), the second trimester (gestational weeks 21-24), and the third trimester (gestational weeks 33-36). Peripheral blood leukocyte and lymphocyte counts were measured in children at 12 months of age using the Coulter principle combined with flow cytometry. Exposure levels of PYRs metabolites in each trimester were divided into low, moderate, and high exposure groups based on the 25th (P₂₅) and 75th (P₇₅) percentiles. Meanwhile, participants were classified as having repeated high or low exposure if their metabolite levels were > P₇₅ or <P₂₅ in at least two trimesters, respectively, while all others were categorized as having repeated moderate exposure. Generalized linear models were used to analyze the associations between trimester-specific and repeated PYRs metabolite exposure levels and the peripheral blood white blood cell (WBC) and lymphocyte counts in children aged 1 year. Results A total of 336 mother-child pairs were included in this study. For the pregnant women, the total detection rates of maternal urinary 3PBA, 4F3PBA, and cis-DBCA across the three trimesters of pregnancy were 80.5%, 100.0%, and 81.3%, respectively; and median creatinine-corrected concentrations were 0.24, 0.36, and 0.42 μg·g⁻¹, respectively. In children aged 1 year, the mean WBC and lymphocyte counts in peripheral blood were (8.9±2.0)×10⁹·L⁻¹ and (5.7±1.6)×10⁹·L⁻¹, respectively. The results of the generalized linear model analysis indicated that compared to the low exposure group, the high cis-DBCA exposure group during the third trimester of pregnancy had significantly lower peripheral blood WBC count (β=−0.87, 95%CI: −1.51, −0.23) and lymphocyte count (β=−0.64, 95%CI: −1.15, −0.13); and the repeated high-exposure group of cis-DBCA had significantly lower peripheral blood WBC count (β=−1.34, 95%CI: −2.34, −0.34) and lymphocyte count (β=−0.80, 95%CI: −1.60, −0.01) than the repeated low exposure group. Similarly, the repeated moderate-exposure group of cis-DBCA had a significantly lower peripheral blood WBC count (β=−0.83, 95%CI: −1.59, −0.07) than the repeated low exposure group. Conclusion High maternal exposure to PYRs with cis-DBCA as the major metabolite exposure is associated with decreased peripheral leukocyte and lymphocyte counts in children aged 1 year, and repeated high-level exposure throughout gestation appears to exacerbate DIT in offspring. The third trimester of pregnancy maybe a sensitive window for children's DIT induced by exposure to PYRs during pregnancy.

OBJECTIVE To investigate the effects and mechanism of Yigan huayu formula in alleviating liver fibrosis in mice. METHODS Mice were randomly divided into blank group （normal saline）， model group （normal saline）， Yigan huayu formula low- and high-dose groups （28.98， 57.96 g/kg， calculated by crude drug）， with 8 mice in each group. Except for the blank group， the liver fibrosis model was induced by intraperitoneal injection of 15%CCl 4 -olive oil solution. From the third week， the mice received the medicine/normal saline intragastrically， once a day， for 4 consecutive weeks. After the last medication， liver indexes were calculated， the activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum， as well as the hydroxyproline （HYP） content in liver tissue， were measured. Liver histopathology was evaluated. Differentially expressed proteins （DEPs） in liver tissue were analyzed based on proteomics， followed by bioinfo rmatics analysis. The expressions of core DEPs were validated using Western blot （WB） and immunohistochemistry （IHC） methods. RESULTS Compared with the blank group， the model group showed significantly elevated liver indexes， serum activities of ALT and AST， and hepatic HYP content （ P ＜0.05）， along with obvious pathological damage and collagen deposition. Compared with the model group， the above indexes of mice in the Yigan huayu formula high-dose group were decreased significantly （ P ＜0.05）， with marked improvement in liver pathological damage and collagen deposition. Proteomics identified 210 DEPs between the model group and Yigan huayu formula high-dose group. DEPs were significantly enriched in extracellular matrix （ECM）-receptor interaction and lipid metabolism pathways. WB and IHC confirmed that Yigan huayu formula could significantly inhibit the abnormally elevated expressions of collagen type Ⅳ alpha1 chain （COL4A1）， secreted protein acidic and rich in cysteine （SPARC）， vitronectin （VTN） and laminin subunit alpha5 （LAMA5） in liver tissue of mice （ P ＜0.05）. CONCLUSIONS Yigan huayu formula may exert anti-hepatic fibrosis effects by inhibiting the expressions of proteins such as COL4A1， LAMA5， SPARC， and VTN， thereby blocking the ECM-receptor interaction signaling pathway， and subsequently suppressing excessive ECM deposition and basement membrane remodeling.

ObjectiveThis paper aims to clarify the material basis of Gualou Niubangtang and establish a quantitative analysis method for its main constituents， providing a reference for the overall quality control of this preparation. MethodsThe constituents in the formula were systematically characterized based on ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS/MS）. Identification was performed by matching with the UNIFI 9.6 software and utilizing database platforms such as PubChem， ChemicalBook， and ChemSpider， combined with relevant literature reports. A quantitative analysis method for the seven main constituents in Gualou Niubangtang was established by using high performance liquid chromatography （HPLC）. ResultsUPLC-Q-TOF-MS/MS analysis identified 155 constituents， including 69 flavonoids， 36 terpenoids， 23 phenylpropanoids， 8 phenylethanoid glycosides， and 19 other types of constituents. In the established quantitative analysis method， the seven main constituents showed good linearity within their respective linear ranges. The precision， repeatability， stability， and spike recovery all met the required standards. The results showed that the content ranges of geniposide， liquiritin， hesperidin， arctiin， baicalin， oroxylin A-7-O-β-D-glucuronide， and wogonoside in 15 batches of Gualou Niubangtang were 13.67-21.25， 1.20-7.64， 5.45-7.45， 22.97-33.51， 29.95-39.07， 2.58-4.80， and 6.56-9.31 mg·g^-1， respectively. ConclusionThis study successfully characterizes and attributes multi-category constituents in Gualou Niubangtang， clarifying that its material basis is primarily composed of flavonoids， terpenoids， phenylethanoid glycosides， and phenylpropanoids. Furthermore， it enables the quantification of seven constituents within the formula. This work lays a foundation for research on the quality control， action mechanism， and clinical application of this formula.

Objective To investigate the protective effect and potential mechanism of kinsenoside （KD）-the main ingredient of Anoectochilus roxburghii （AR） on alcoholic liver injury in mouse models of chronic and acute alcoholic liver injury, and provide a theoretical basis for the development of drugs for alcoholic liver injury. Methods Chronic and acute alcoholic liver injury mouse models were induced by feeding liquid diet containing 30% alcohol and gavage of high doses of alcohol （6 g/kg）, respectively. The KD （50 mg/kg） and AR （250 mg/kg） were administrated by intragastric administration. Body weight, liver index, serum alanine aminotransferase （ALT）, aspartate aminotransferase （AST） levels, serum total triglyceride （TG） and cholesterol （TC） levels were measured; hematoxylin-eosin and oil red O staining was performed on liver tissues; lipid metabolic related genes （PPARα and SREBP1） expression levels were detected by QPCR. Results Both models of alcoholic liver injury resulted in increased hepatic transaminase activity and elevated lipids, accompanied by massive vacuolar structure and lipid droplet formation in pathological liver sections. In the chronic alcoholic liver injury model, ALT and AST were significantly reduced after KD or AR treatment （P<0.05, P<0.001）; the transcriptional activity of SREBP1 was significantly reduced after KD or AR treatment （P<0.01, P<0.05）. In the acute alcoholic liver injury model, AST was significantly reduced after KD or AR treatment （P<0.01, P<0.01）, and TG level was significantly decreased （P<0.01, P<0.01）. Conclusion KD, as the main active ingredient of AR, played a major role in hepatoprotection in mice. KD treatment significantly alleviated chronic and acute alcoholic liver injury and reduced the lipid deposition in liver; KD promoted lipolysis by increasing PPARα and inhibiting the expression of SREBP1 to reduce the synthesis and accumulation of lipids, thus exerting its role in regulating lipid metabolism, which suggested that KD, as the active ingredient of AR, could be a potential drug for the treatment of ALD.

Immunoglobulin G4-related disease (IgG4-RD) is a rare systemic immune-mediated disorder characterized by infiltration of IgG4-positive plasma cells and multiorgan fibrosis. Among them, cases with multiple pancreatic cystic lesions as the prominent manifestation are relatively rare, and they are easily misdiagnosed as pancreatic tumors or pancreatitis in clinical practice. Herein, we report one patient with IgG4-RD admitted to Peking Union Medical College Hospital. The patient initially presented with multiple pancreatic cystic lesions and aneurysmal dilatation of the abdominal aorta, accompanied by markedly elevated serum IgG4 levels. Combined with pathological findings from submandibular gland biopsy, the diagnosis of IgG4-RD was ultimately established. After treatment with a regimen of glucocorticoids combined with cyclophosphamide, the patient′s condition improved. Meanwhile, we performed a literature review of cases with IgG4-RD-related pancreatic cystic lesions, aiming to improve clinicians′ understanding of this rare disease phenotype.

Objective To explore the relationship and predictive value between serum neurofilament light chain protein(NfL),endothelial cell specific molecule-1(Endocan),B cell activating factor(BAFF)and epi-lepsy secondary to acute cerebral infarction(ACI).Methods From June 2020 to June 2023,135 patients with secondary epilepsy due to ACI admitted to the hospital were included in the epilepsy group,and 90 ACI pa-tients without secondary epilepsy admitted during the same period were included in the non-epilepsy group.The general data of the two groups and the levels of serum NfL,Endocan and BAFF were determined and compared.Multivariate Logistic regression was used to analyze the related influencing factors of epilepsy sec-ondary to ACI.The receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum NfL,Endocan and BAFF for epilepsy secondary to ACI.Results Compared with non-epilepsy group,the age,the proportion of infarction lesions involving cortex,the proportion of National Institutes of Health Stroke Scale(NISSH)score≥15 points,the proportion of infarct volume＞10 cm3,the proportion of family history of epilepsy,and serum NfL,Endocan and BAFF were significantly increased in epileptic group(P＜0.05).Among patients with infarction lesions involving cortex and NISSH score ≥15 points,the expression levels of serum NfL,Endocan,and BAFF were significantly higher in those with epilepsy than in those without epilepsy(P＜0.05).Multivariate Logistic regression analysis showed that infarction involving the cortex,NISSH score ≥15 points,elevated serum NfL,Endocan and BAFF were risk factors for secondary epilepsy in ACI(P＜0.05).The results of ROC curve analysis showed that the area under the curve and specificity of the combined prediction of serum NfL,Endocan,and BAFF for secondary epilepsy in ACI were superior to those of individual detection or the combined detection of the two(P＜0.05).Conclusion The high expression of serum NfL,Endocan and BAFF are risk factors for secondary epilepsy in ACI.The combined detection of the three has a high predictive value for secondary epilepsy in ACI.

Objective:To evaluate the efficacy and safety of IBUCy(idarubicin,busulfan,and cyclophosphamide)and FABC(fludarabine,cyta-rabine,busulfan,and cyclophosphamide)conditioning regimens followed by allogeneic hematopoietic stem cell transplantation(allo-HSCT)for the treatment of medium-to-high risk acute myelocytic leukemia(AML).Methods:We retrospectively analyzed data of 49 patients with medium-to-high risk AML who received IBUCy(n=17)or FABC(n=32)conditioning regimens followed by allo-HSCT between January 2015 and December 2021 at The First Affiliated Hospital of Chongqing Medical University.Hematopoietic reconstruction time,adverse events,and survival outcomes were compared between the two groups to assess the efficacy and safety of the two regimens.Additionally,we analyzed factors that may be associated with prognosis.Results:Hematopoietic reconstruction was successful in all 49 patients.No significant differ-ences were observed between the two groups in terms of hematopoietic reconstruction time.Similarly,no significant differences were ob-served in the 5-year progression-free survival(PFS)and overall survival(OS)rates between the two groups.The incidence rates of oral mu-cositis,nausea and vomiting,diarrhea(≥grade 3 based on CTCAE v5.0),and chronic graft-versus-host disease(GVHD)were significantly high-er in the IBUCy group than that in the FABC group.However,the incidence rate of hemorrhagic cystitis in the FABC group was significantly higher than that in the IBUCy group.The time from diagnosis to allo-HSCT>6 months and being minimal residual disease(MRD)-positive be-fore transplantation were identified as the risk factors for PFS(P=0.019 and P=0.048,respectively).Patients who were MRD-negative before transplantation had significantly longer PFS when treated with the IBUCy conditioning regimen(P=0.039).Conclusions:Both IBUCy and FABC conditioning regimens prior to allo-HSCT are safe and effective for treating medium-to-high risk AML.Allo-HSCT should be performed as soon as possible when patients achieve their first complete remission.Patients with an MRD-negative status before transplantation tend to have longer PFS.Compared with the FABC regimen,the IBUCy regimen has some advantages;however,attention should be given to the prevention and management of gastrointestinal adverse events and chronic GVHD.

OBJECTIVE: To investigate the effects of bortezomib (BTZ) combined with polyphyllin Ⅶ (PP7) on proliferation, apoptosis and oxidative stress of myeloma cell line ARH-77. METHODS: MTT assay was used to detect the inhibitory effects of different concentrations of BTZ, PP7 monotherapy, and their combination on the proliferation of ARH-77 cells. In subsequent experiments, the cells were divided into 4 groups: control group (no drug added), BTZ (15 nmol/L) group, PP7 (1.5 μmol/L) group and BTZ(15 nmol/L)+PP7 (1.5 μmol/L) group. The effects of the two drugs on the morphology of ARH-77 cells were observed. Flow cytometry was used to detect the apoptosis rate of the cells in each group. Calcein-AM/PI double staining kit was used to observe the status of the cells and the cell viability were evaluated. The expression of apoptosis-related proteins were detected by Western blot. DCFH-DA fluorescent probe was used to detect the levels of reactive oxygen species (ROS). RESULTS: Both BTZ and PP7 monotherapy, as well as their combination, could inhibit the growth of ARH-77 cells in a dose-dependent manner (r_BTZ=-0.9717, r_PP7=-0.9941, r_BTZ+PP7=-0.9951), and the combination of BTZ and PP7 exhibited a synergistic effect within a certain concentration range. Compared with the BTZ group and PP7 group, the apoptosis rate of the BTZ+PP7 group was significantly increased (P < 0.01), the expressions of pro-apoptotic proteins Bax, Smac and P53 were significantly upregulated (P < 0.05), the expression of anti-apoptotic protein Bcl-2 was significantly downregulated (P < 0.01), and the ratio of Bax/Bcl-2 was significantly increased (P < 0.01). Compared with the control group, the level of ROS in the BTZ, PP7 monotherapy group and BTZ+PP7 group were significantly increased (P < 0.05). CONCLUSION BTZ combined with PP7 can inhibit the proliferation and induce apoptosis of ARH-77 cells, and increase the level of intracellular ROS.
Apoptosis/drug effects* ; Bortezomib ; Humans ; Cell Proliferation/drug effects* ; Oxidative Stress/drug effects* ; Multiple Myeloma/metabolism* ; Cell Line, Tumor ; Saponins/pharmacology*