Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.

Objective:To evaluate the relationship between hippocampal receptor-interacting protein kinase-1 (RIPK1) and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasomes in postoperative neurocognitive dysfunction of aged rats with chronic knee arthritis pain.Methods:Sixty-four healthy male Sprague-Dawley rats, aged 18 months, weighing 500-550 g, were divided into 4 groups ( n=16 each) using a random number table method: chronic knee arthritis pain group (group P), chronic knee arthritis pain+ operation group (group PS), RIPK1 inhibitor necrostatin-1+ chronic knee arthritis pain+ operation group (group NPS), and DMSO+ chronic knee arthritis pain+ operation group (group DPS). The knee arthritis model was prepared by intra-articular injection of monosodium iodoacetate (MIA) 1 mg into the left knee joint, and 12 weeks later exploratory laparotomy was performed under sevoflurane anesthesia. Necrostatin-1 6.25 mg/kg and the equal volume of DMSO were intraperitoneally injected at 1 h before operation in NPS group and DPS group, respectively. Thermal pain threshold was measured at 1 week before MIA injection and 6 and 12 weeks after MIA injection. Morris water maze test was used to evaluate the cognitive function at 7 days after surgery. Hippocampal tissues were obtained for microscopic examination of the pathological changes (after HE staining) and for determination of the expression of RIPK1, phosphorylated RIPK1 (p-RIPK1), NLRP3, activated cysteine-aspartic protease caspase-1 (cl-caspase-1), apoptosis-associated speck-like protein containing a CARD (ASC) (by Western blot) and contents of interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results:Thermal pain threshold was significantly decreased at 6 and 12 weeks after MIA injection as compared with that before injection ( P<0.05), and there was no significant difference in thermal pain threshold among the four groups ( P>0.05). Compared with P group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the expression of RIPK1, p-RIPK1, NLRP3, cl-caspase-1 and ASC was up-regulated, and the contents of IL-1β and IL-18 were increased ( P<0.05), and pathological changes of hippocampal neurons were marked in PS group, DPS group and NPS group. Compared with PS group and DPS group, the escape latency was significantly shortened, the time of staying at the original platform quadrant was prolonged, the number of crossing the original platform was increased, the expression of RIPK1, p-RIPK1, NLRP3, cl-caspase-1 and ASC was down-regulated, the contents of IL-1β and IL-18 were decreased ( P<0.05), and pathological changes of hippocampal neurons were significantly attenuated in NPS group. Conclusions:Postoperative hippocampal RIPK1 function is enhanced in aged rats with chronic knee arthritis pain, which then activates NLRP3 inflammasomes, triggering neuroinflammation, and this process may be involved in the mechanism of postoperative neurocognitive dysfunction.

AIM To explore the effects of Buyang Huanwu Decoction on mitochondrial oxidative damage and PKCε-Nampt pathway in rats following cerebral ischemia reperfusion(I/R).METHODS The rats were randomly divided into the sham operation group,the model group,Buyang Huanwu Decoction group(14.3 g/kg)and edaravone group(3 mg/kg).Except those of the sham operation group,SD rats of other groups were induced into models of brain I/R injury by MCAO method,followed by corresponding drug administration 24 hours after operation.After 7 days of administration,the rats had their neurological deficit evaluated by neurological function scoring;thier expression of neuron marker MAP-2 detected by immunofluorescence staining;their neuron damage observed and the oxidative damage evaluated through assessment of their ROS levels and MDA and SOD activities;their changes of mitochondrial membrane potential detected by fluorescent probe JC-1;their ratio of NAD+/NADH detected using modified enzyme circulation method;their expressions of PKCε,p-PKCε and Nampt proteins detected with Western blot;and their positive expressions of p-PKCε and Nampt proteins detected with immunohistochemistry method.RESULTS Compared with the model group,Buyang Huanwu Decoction group shared decreased cerebral infarction volume and neurological function score(P<0.05);increased cerebral fluorescence intensity of MAP-2(P<0.05);reduced neuronal damage,decreased cerebral levels of ROS and MDA(P<0.05);increased SOD activity,mitochondrial membrane potential and NAD+/NADH ratio(P<0.05);and increased protein expressions of p-PKCε and Nampt(P<0.05).CONCLUSION Buyang Huanwu Decoction can improve mitochondrial function and reduce brain I/R injury in rats by activating their PKCε-Nampt signaling pathway.

Objective To analyze the clinical manifestations of delayed serological transfusion reactions(DSTR)after platelet transfusion in tumor patients,and to explore the transfusion strategy.Methods Clinical data and laboratory test re-sults of patients with positive antibody screening were analyzed after platelet transfusion in our hospital from January 1,2015 to June 30,2023,and the incidence rate,clinical characteristics and transfusion strategy of patients with DSTR were ana-lyzed.Results A total of 2 553 patients with 6 057 platelet transfusions were reviewed.Eight patients developed DSTR and received a total of 21 therapeutic amounts of platelets,and 5 patients were subsequently transfused with red blood cells.Rh system antibodies were detected in 7 cases(4 anti-E,1 anti-c/E,1 anti-C and 1 anti-c)and Kell system antibodies in 1 case.Conclusion Tumor patients may also develop DSTR after platelet transfusion.It is necessary to pay close attention to the antibody situation and perform matched transfusion when transfusing blood again.

Objective:To investigate the association between congenital hypothyroidism (CH) and the adverse outcomes during hospitalization in very low birth weight infants (VLBWI).Methods:This prospective, multicenter observational cohort study was conducted based on the data from the Sino-northern Neonatal Network (SNN). Data of 5 818 VLBWI with birth weight <1 500 g and gestational age between 24-<37 weeks that were admitted to the 37 neonatal intensive care units from January 1 st, 2019 to December 31 st, 2022 were collected and analyzed. Thyroid function was first screened at 7 to 10 days after birth, followed by weekly tests within the first 4 weeks, and retested at 36 weeks of corrected gestational age or before discharge. The VLBWI were assigned to the CH group or non-CH group. Chi-square test, Fisher exact probability method, Wilcoxon rank sum test, univariate and multivariate Logistic regression were used to analyze the relationship between CH and poor prognosis during hospitalization in VLBWI. Results:A total of 5 818 eligible VLBWI were enrolled, with 2 982 (51.3%) males and the gestational age of 30 (29, 31) weeks. The incidence of CH was 5.5% (319 VLBWI). Among the CH group, only 121 VLBWI (37.9%) were diagnosed at the first screening. Univariate Logistic regression analysis showed that CH was associated with increased incidence of extrauterine growth retardation (EUGR) ( OR=1.31(1.04-1.64), P<0.05) and retinopathy of prematurity (ROP) of stage Ⅲ and above ( OR=1.74(1.11-2.75), P<0.05). However, multivariate Logistic regression analysis showed no significant correlation between CH and EUGR, moderate to severe bronchopulmonary dysplasia, grade Ⅲ to Ⅳ intraventricular hemorrhage, neonatal necrotizing enterocolitis in stage Ⅱ or above, and ROP in stage Ⅲ or above ( OR=1.04 (0.81-1.33), 0.79 (0.54-1.15), 1.15 (0.58-2.26), 1.43 (0.81-2.53), 1.12 (0.70-1.80), all P>0.05). Conclusion:There is no significant correlation between CH and in-hospital adverse outcomes, possibly due to timely diagnosis and active replacement therapy.

Background and objective Tumor microenvironment(TME)is one of the important factors in tu-morigenesis and progression,in which tumor-associated macrophages(TAMs)play an important role in non-small cell lung cancer(NSCLC)progression.However,the mechanism of TAMs in NSCLC progression remains unclear,so this study aimed to investigate the role of TAMs in NSCLC progression and to find potential therapeutic targets.Methods Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze the expression of prostaglandin E2 receptor 4(EP4)mRNA in NSCLC and normal lung tissues;the protein expression levels of cyclooxygenase-2(COX-2),EP4,cluster of differentiation 86(CD86),CD163 and CD31 were detected by immunohistochemistry(IHC)in 120 NSCLC tissues and 24 paracancerous tissues specimens.The nude mouse lung adenocarcinoma cell A549 and macrophage RAW264.7 co-transplanted tumor model was established.And the samples were collected by gavage with EP4 inhibitor E7046,and then stained with hematoxylin-eosin(HE),IHC,and immunofluorescence(IF),and then detected by Western blot for the epithelial mesenchymal transformation(EMT)of the tumor tissues of the nude mice in each group.Western blot was used to detect the expressions of EMT related protiens in each group of nude mice;full-length transcriptome sequencing was used to screen the key genes causing liver me-tastasis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was performed.Results EP4 mRNA expression level in NSCLC tissues was generally lower than that in normal lung tissues(P<0.05);COX-2,EP4,CD163,CD31 proteins were differentially expressed in NSCLC tissues and adjacent tissues,and differences were observed in many clinico-pathological parameters of NSCLC patients;RAW264.7 shortened the latency period of tumorigenesis of A549 and promoted the proliferation of tumors and liver metastasis of tumors,and E7046 could reduce tumor cell proliferation activity,tumor tissue vascular density and M2-type macrophage infiltration in nude mice;IF staining showed that macrophages were mainly distributed around the metastatic foci of tumors;Western blot results showed that compared with A549 alone transplantation group,the relative expression of E-cadherin protein in tumor tissues of mice in A549 and RAW264.7 co-transplantation group was significantly decreased,and the difference was statistically significant(P<0.05),while the relative expression of N-cadherin protein was up-regulated,but the difference was not statistically significant(P>0.05);the main pathways enriched in the differ-ential genes of the full-length transcriptome were the PI3K-AKT and MAPK signaling pathways.Conclusion During NSCLC development,the COX-2/PGE2/EP4 axis may promote tumor progression by inducing macrophage functional activation,and EP4 may be a potential new target for tumor immunotherapy.This study provides new perspectives and ideas for in-depth exploration of the mechanisms of NSCLC development,as well as a theoretical basis for the development of new therapeutic strategies for NSCLC.

Objective:to analyze the relationship between the changes of lymphocyte subsets and cardiac function classification in elderly patients with chronic heart failure and preserved ejection fraction.Methods:A total of 835 elderly patients with preserved ejection fraction treated in our hospital from August 2022 to June 2023 were retrospectively selected, and divided into grade Ⅰ(302 cases), grade Ⅱ(254 cases), grade Ⅲ(144 cases)and grade Ⅳ(135 cases)according to the cardiac function classification, and 211 cases in the same period were selected as the control group.The general data of patients were collected, and the expression of T lymphocyte subsets and serum levels of cardiac related factor in patients with different cardiac function classifications were compared and analyzed.Results:There were no statistically significant differences in age, gender, BMI, systolic blood pressure, diastolic blood pressure and heart rate between the two groups( P>0.05).The lower the cardiac function classification, the higher the levels of CD4 + [grade Ⅰ: (39.7±8.0)%, grade Ⅱ: (36.9±8.3)%, grade Ⅲ: (32.1±6.5)%, grade Ⅳ: (31.3±5.1)%], CD4 + /CD8 + [grade Ⅰ: (1.8±0.5), grade Ⅱ: (1.5±0.5), grade Ⅲ: (1.4±0.5), grade Ⅳ: (1.1±0.3)], and the lower the levels of CD8 + [grade Ⅰ: (23.9±5.1)%, grade Ⅱ: (25.5±8.4)%, grade Ⅲ: (26.3±10.0)%, grade Ⅳ: (30.7±9.0)%]in T lymphocyte subsets( F=56.846, 84.154, 23.965, P<0.05); the lower the levels of sST2, GAL-3 and GDF-15 in serum of patients with lower cardiac function classification( F=217.081, 141.741, 123.835, P<0.05); CD4 + and CD4 + /CD8 + were negatively correlated with cardiac function grading( r=-0.482 and r=-0.849, P<0.05), CD8 + was positively correlated with cardiac function grading( r=0.948, P<0.05); the effect of using T lymphocyte subsets to diagnose cardiac function grading was higher and the AUC was 0.984. Conclusions:The accuracy of lymphocyte subsets in the diagnosis of cardiac function grading in elderly patients with chronic heart failure and preserved ejection fraction is higher, which is worthy of promotion.

Objective:To analyze the factors influencing sarcopenia in older patients with chronic diseases, both in community settings and hospitals, and to develop a risk prediction model for sarcopenia.Methods:We recruited a total of 403 older adults with chronic diseases, consisting of 251 individuals from a community in Nanjing, Jiangsu Province, and 152 hospitalized patients from the Department of Geriatrics at Jiangsu Province Hospital.Assessments were conducted using a general information questionnaire, serum sample collection, the mini nutritional assessment-short form(MNA-SF), the mini-mental state examination(MMSE), and the geriatric depression scale(GDS).Binary Logistic regression analysis was employed to identify influencing factors and to construct a risk prediction model for sarcopenia, which was illustrated using a nomogram.The model's discrimination was evaluated using the receiver operating characteristic(ROC)curve and the area under the curve(AUC).Results:The prevalence of sarcopenia among community-dwelling older adults with chronic conditions was found to be 4.0%(10/251).In contrast, the prevalence in hospitalized older adults with chronic conditions was significantly higher at 36.2%(55/152).Binary Logistic regression analysis identified several independent risk factors for sarcopenia, including hospitalization( OR=14.391、95% CI: 6.284-32.955、 P<0.001), male gender( OR=3.321、95% CI: 1.587-6.950、 P=0.001), lower low-density lipoprotein cholesterol(LDL-C)levels( OR=2.542、95% CI: 1.160-5.572、 P=0.020), cognitive impairment( OR=2.654、95% CI: 1.269-5.550、 P=0.010), and the use of four or more types of medication( OR=2.328、95% CI: 1.952-5.689、 P=0.044).Based on these risk factors, a nomogram was developed as a predictive model for assessing sarcopenia risk.The AUC for this prediction model was 0.860(95% CI: 0.815-0.912), indicating a sensitivity of 0.831 and a specificity of 0.760. Conclusions:The incidence of sarcopenia is notably high among older patients with chronic diseases.A risk prediction model that incorporates factors such as hospitalization history, gender, LDL-C levels, cognitive function, and types of medication demonstrates significant potential for predicting sarcopenia.This model serves as a valuable foundation for the early screening and intervention of sarcopenia.

italic>Platycodon grandiflorum (Jacq.) A. DC is one of the most commonly used bulk medicinal herbs. It has important value in the fields of medicine, food and cosmetics, and its market demand is increasing year by year, and it has a good development prospect. In this study, based on 403 distribution records and 8 environmental variables, we used Maxent model to predict the potential distribution of P. grandiflorum under climate change. The results showed that the model simulation effect was the best when the Maxent parameter was FC (feature combination) = LQPH (L: linear features; Q: quadratic features; P: product features; H: hinge features) and RM (regularization multiplier) = 2.1, AUC (area under curve) = 0.901, and the model prediction showed that P. grandiflorum was widely distributed in 28 provinces of China under the current climate conditions, with a suitable area of 2 337 419.98 km². Under the influence of climate change in the future, the area of suitable habitat of P. grandiflorum showed a decreasing trend, and the distribution center as a whole showed a trend of northward and eastward shift. The distribution area of P. grandiflorum in the three main producing areas is affected by climate change, and the overall trend is that the future suitable distribution area of Chifeng in Inner Mongolia increases, while the future suitable distribution area of Zibo in Shandong and Taihe, Bozhou in Anhui decreases or even disappears under the SSP5-8.5 scenario (shared socioeconomic pathways). It is suggested to maintain and protect the ecological environment and germplasm resources of the reserved area suitable for the growth of P. grandiflorum, and increase the cultivation research in the expansion area of P. grandiflorum, so as to lay a foundation for the subsequent expansion of P. grandiflorum planting and to promote the protection and sustainable development of P. grandiflorum resources.

Pharmaceutical cocrystals is an advanced technology to improve the physicochemical and biological properties of drugs. However, there are few studies on the in vivo metabolism of pharmaceutical cocrystals. In this study, the pharmacokinetics of wogonin cocrystal in normal rats was further studied on the basis of the previous preparation of wogonin-aloperine cocrystal. Firstly, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for simultaneous determination of wogonin and its metabolite wogonoside in rat plasma, and to investigate the methodology. The method was applied to the pharmacokinetic study of wogonin-aloperine cocrystal (Wog-Alop) in rats. The results showed that wogonin and its metabolite wogonoside had a good linear relationship in the range of 1-800 ng·mL^-1, and the precision, accuracy, matrix effect and stability in this range met the requirements of biological analysis. Compared with direct administration of wogonin, the C_max of wogonin and its metabolite in rats increased to 7.44-fold and 9.15-fold, AUC_0-t increased to 1.67-fold and 3.72-fold, and oral bioavailability of wogonin increased to 187.66% after cocrystal administration. Wog-Alop cocrystal can significantly improve the C_max, AUC, and oral bioavailability of wogonin and its metabolite, which provides a new perspective for the clinical application of wogonin. This study was approved by the Experimental Animal Ethics Review Committee of Beijing University of Chinese Medicine (approval number: BUCM-2023032307-1148).