OBJECTIVE To explore the mechanism of joint injection of Caulophyllum robustum Maxim in the treatment of rheumatoid arthritis （RA）. METHODS The targets of main saponins in C. robustum Maxim were obtained from Swiss Target Prediction， and the RA treatment targets collected from the GeneCards and OMIM database were intercrossed to establish an interaction network based on network pharmacology. Gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis were performed. RA model was established by injecting complete Freund’s adjuvant into the back of rabbits for verification. The arthritis index score， knee diameter and pain threshold of rabbits were compared. Pathological examination of rabbit synovial tissue was carried out. The levels of tumor necrosis factor α （TNF-α）， interleukin-1β （IL-1β） and IL-6 in rabbit serum and synovial fluid were detected. The phosphorylation levels of tyrosine protein Janus kinase 2 （JAK2） and signal transducer and activator of transcription 3 （STAT3） proteins in rabbit synovium were detected. RESULTS Network pharmacology identified 143 intersection targets between the drug and RA. After the construction of the “drug-component-target” network， the core components of the network were echinocystic acid， oleanolic acid， hederagenin， cauloside A and cauloside C， etc. Additionally， the top 10 core targets of PPI network were SRC， STAT3， MAPK1， EGFR， PIK3CA， MAPK3， GRB2， JUN， PTPN11 and JAK2. The results of KEGG pathway analysis showed that the JAK/STAT signaling pathway was mainly involved in the treatment of RA by joint injection of C. robustum Maxim. Results of validation test showed that compared with model group， joint injection of C. robustum Maxim could reduce the swelling of rabbit knee joint， relieve the hyperplasia of synovial layer， reduce the hyperplasia of lower connective tissue， and reduce the number of inflammatory cells and capillaries. The arthritis index score （excluding low-dose group of C. robustum Maxim）， knee diameter， the levels of TNF-α， IL-1β and IL-6 in serum and synovial fluid， and the protein phosphorylation levels of JAK2 and STAT3 were decreased significantly （P＜0.05 of P＜0.01）， while the pain threshold were reduced significantly （P＜0.01）. CONCLUSIONS The core components that may alleviate the inflammatory response of RA in joint injection of C. robustum Maxim could include echinocystic acid， oleanolic acid， hederagenin， cauloside A， and cauloside C. Its mechanism may be related to the inhibition of JAK/STAT signaling pathway and the reduction of inflammatory responses.

Background Manganese is an essential trace element for the human body and maintains normal development of many organs including the brain. However, long-term exposure to a high manganese environment or excessive manganese intake will lead to manganese poisoning and result in neurological diseases, and currently no effective treatment plan is available. Objective To develop an animal model for subchronic manganese exposure and assess the impact of Dendrobium nobile Lindl. alkaloids (DNLA) on manganese associated behavioral and hippocampal effects in rats. Methods Fifty male SPF SD rats were randomly allocated into a control group (0.9% normal saline by intraperitoneal injection), two experimental groups [7.5 mg·kg⁻¹ (low) or 15 mg·kg⁻¹ (high) of MnCl₂·4H₂O by intraperitoneal injection], and two DNLA antagonistic groups [15 mg·kg⁻¹ MnCl₂·4H₂O by intraperitoneal injection then either 20 mg·kg⁻¹ (low) or 40 mg·kg⁻¹ (high) DNLA by oral administration]. All groups of rats were adminaistered 5 d per wek, once a day, for consecutive 13 weeks. Following modeling, neurobehavioral assessments were conducted using open field, Morris water maze, and Y maze. Inductively coupled plasma mass spectrometry (ICP-MS) was utilized to measure manganese levels in the blood and brain tissues of the rats, and hematoxylin-eosin (HE) staining was employed to examine neuronal morphological changes in the hippocampal tissues of the rats. Results The neurobehavioral tests revealed that the manganese-exposed rats exhibited decreased total movement distance, prolonged central zone dwelling time, and reduced motor activity in the open field test, indicating tendencies toward depression and anxiety (P<0.05). In the Y-maze test, the mean exploration distance in the novel arm, the number of entries into the novel arm, and the time spent in the novel arm of the managanses-exposed rats were all reduced, while the latency period increased, suggesting impaired spatial exploration and learning-memory functions (P<0.05). In the Morris water maze navigation test, the escape latency was significantly longer in the manganese-exposed rats compared to the control group, and the number of platform crossings decreased in the spatial probe test, indicating a significant decline in spatial learning and memory (P<0.05). The ICP-MS analysis showed elevated manganese concentrations in the blood and hippocampus of the exposed rats (P<0.05), and the histopathological observation revealed hippocampal damage. Following the DNLA intervention, the manganese-exposed rats showed increased total movement distance and reduced central zone dwelling time in the open field test (P<0.05). In the Y-maze test, the mean exploration distance in the novel arm, the number of entries into the novel arm, and the time spent in the novel arm increased, while the latency period decreased, suggesting alleviation of anxiety and improved exploratory behavior (P<0.05). In the Morris water maze test, the escape latency gradually shortened, and both the number of platform crossings and the percentage of time spent in the target quadrant increased, indicating improved spatial learning and memory (P<0.05). Additionally, the manganese levels in the blood and hippocampus decreased (P<0.05), and the hippocampal pathological changes were partially restored. Conclusion DNLA demonstrates the ability to counteract multiple neurotoxic effects following the elevation of manganese levels in the blood and hippocampal tissues of rats induced by subchronic manganese exposure. Specifically, DNLA is shown to ameliorate the behavioral alterations observed in rats after manganese exposure, and mitigate the hippocampal damage in manganese-exposed rats.

Successful cloning through somatic cell nuclear transfer (SCNT) faces significant challenges due to epigenetic obstacles. Recent studies have highlighted the roles of H3K4me3 and H3K27me3 as potential contributors to these obstacles. However, the underlying mechanisms remain largely unclear. In this study, we generated genome-wide maps of H3K4me3 and H3K27me3 in mouse pre-implantation NT embryos. Our analysis revealed that aberrantly over-represented broad H3K4me3 domain and H3K27me3 signal lead to increased bivalent marks at gene promoters in NT embryos compared with naturally fertilized (NF) embryos at the 2-cell stage, which may link to relatively low levels of H3K36me3 in NT 2-cell embryos. Notably, the overexpression of Setd2, a H3K36me3 methyltransferase, successfully restored multiple epigenetic marks, including H3K36me3, H3K4me3, and H3K27me3. In addition, it reinstated the expression levels of ZGA-related genes by reestablishing H3K36me3 at gene body regions, which excluded H3K27me3 from bivalent promoters, ultimately improving cloning efficiency. These findings highlight the excessive bivalent state at gene promoters as a potent barrier and emphasize the removal of these barriers as a promising approach for achieving higher cloning efficiency.
Animals ; Mice ; Histone-Lysine N-Methyltransferase/biosynthesis* ; Histones/genetics* ; Nuclear Transfer Techniques ; Female ; Gene Expression Regulation, Developmental ; Promoter Regions, Genetic ; Epigenesis, Genetic ; Embryo, Mammalian/metabolism*

OBJECTIVE: To evaluate the impact of critical care warning platform (CWP) on clinical outcomes of patients transferred from internal medical ward to intensive care unit (ICU) based on real-world data. METHODS: A retrospective cohort study was conducted. The patients transferred from internal medical ward to ICU of Zhongda Hospital, Southeast University, between January 2022 and October 2024, were enrolled. They were divided into critical care warning group and conventional treatment group based on whether they were connected to the CWP. The patients in the critical care warning group were connected to the CWP, which collected real-time vital signs and treatment data. The platform automatically calculated severity scores, generated individualized risk assessments, and triggered warning alerts, allowing clinicians to adjust treatment plans accordingly. The patients in the conventional treatment group were not connected to the CWP and relied on conventional clinical judgment and nursing measures for treatment management. Baseline characteristics [gender, age, body mass index (BMI), admission type, severity score of illness, underlying diseases, and disease type at ICU admission], primary clinical outcome (in-hospital mortality), and secondary clinical outcomes [ICU mortality, length of ICU stay, total length of hospital stay, and mechanical ventilation and continuous renal replacement therapy (CRRT) status] were collected. Multivariate Logistic regression was used to analyze the impact of CWP on in-hospital death, and subgroup analyses were performed based on different patient characteristics. RESULTS: A total of 1 281 patients were enrolled, with 768 in the critical care warning group and 513 in the conventional treatment group. Compared with the conventional treatment group, the proportion of patients in the critical care warning group with underlying diseases of diabetes and malignancy and transferred to ICU due to sepsis was lowered, however, there were no statistically significant differences in other baseline characteristics between the two groups. Regarding the primary clinical outcome, the in-hospital mortality in the critical care warning group was significantly lower than that in the conventional treatment group [17.6% (135/768) vs. 25.7% (132/513), P < 0.01]. For secondary clinical outcomes, compared with the conventional treatment group, the patients in the critical care warning group had significantly fewer days of mechanical ventilation within 28 days [days: 2 (1, 6) vs. 2 (1, 8), P < 0.05], significantly shorter length of ICU stay [days: 3 (2, 8) vs. 4 (2, 10), P < 0.01], and significantly lower ICU mortality [15.1% (116/768) vs. 21.4% (110/513), P < 0.01]. Multivariate Logistic regression analysis showed that, after adjusting for age and underlying diseases, the use of CWP was significantly associated with a reduction of in-hospital mortality among patients transferred from internal medical ward to ICU [odds ratio (OR) = 0.670, 95% confidence interval (95%CI) was 0.502-0.894, P = 0.006]. Further subgroup analysis revealed that, among patients transferred to ICU due to sepsis, the use of CWP significantly reduced in-hospital mortality (OR = 0.514, 95%CI was 0.367-0.722, P < 0.001). In patients aged ≥ 70 years old (OR = 0.587, 95%CI was 0.415-0.831, P = 0.003) and those with underlying diseases of malignancy (OR = 0.124, 95%CI was 0.046-0.330, P < 0.001), CWP also showed significant protective effects on in-hospital prognosis. CONCLUSION The use of CWP is significantly associated with a reduction in in-hospital mortality among patients transferred from internal medical ward to ICU, demonstrating its potential in assessing the deterioration of hospitalized patients.
Humans ; Intensive Care Units ; Retrospective Studies ; Hospital Mortality ; Prognosis ; Critical Care ; Male ; Female ; Patient Transfer ; Middle Aged ; Aged ; Cohort Studies

Objective Pertussis cases have increased markedly since 2018 in Guangxi.The aim of this study was to evaluate antibody levels and the infection status of pertussis in the resident population. Method A total of 10,215 serum samples from residents were collected from August-November 2018 and tested for anti-pertussis IgG and toxin IgG using the enzyme-linked immunosorbent assay(ELISA). Results Of the collected samples,1,833(17.94%)tested positive for anti-pertussis IgG,with the median concentration of 16.06 IU/mL.Antibody level<10 IU/mL accounted for more than 60%in children under 4 years of age,but declined with age,whereas the percentages of the other three levels(10-40,40-50,and≥50 IU/mL)increased almost with age(P<0.001).Moreover,7,924 samples were selected for anti-pertussis toxin IgG,of which 653(8.24%)tested positive(≥40 IU/mL)with the median concentration of 5.89 IU/mL,and 204 participants(2.56%)had recent pertussis infection(≥100 IU/mL).Among the different age groups,the highest rates of positivity and recent infection were observed at 11-20 years of age,the lowest positivity rate at 5 years of age,and the lowest recent infection rate at 4 years of age(P<0.001,P=0.005,respectively). Conclusion The survey results showed that all age groups in Guangxi lacked immunity against pertussis,which was one of the main factors contributing to the resurgence of pertussis in 2018.In addition,the prevalence of pertussis is relatively high in Guangxi,and its incidence is seriously underestimated,especially in adolescents and adults.

Objective To investigate the effect of the protein kinase RNA-like endoplasmic reticulum kinase(PERK)/eukaryotic initiation factor 2α(eIF2α)pathway in endoplasmic reticulum stress on the activation of hepatic stellate cells(HSC).Methods Pathological sections of normal liver tissue after surgery were collected from 11 patients with hepatic fibrosis(S1-S4)and 9 patients with hepatic hemangioma and hepatic adenoma confirmed by liver biopsy,and immunohistochemistry was used to measure the protein expression levels of PERK,eIF2α,and C/EBP homologous protein(CHOP).Human HSC-LX2 cells were treated with different concentrations of the endoplasmic reticulum stress inducer thapsigargin(0,125,250,500,and 1 000 nmol/L),and qRT-PCR was used to measure the mRNA expression level of PERK,while Western blot was used to measure the protein expression levels of PERK,inositol requiring protein 1(IRE1),activating transcription factor 6(ATF6),CHOP,and α-smooth muscle actin(α-SMA).The method of lentivirus transfection was used to construct a PERK stable overexpression LX-2 group and a control group;qRT-PCR was used to measure the mRNA expression levels of PERK,eIF2α,and α-SMA,Western blot was used to measure the protein expression levels of PERK,phosphorylated eIF2α(p-eIF2α),and α-SMA,and immunofluorescence assay was used to measure the expression of collagen type Ⅰ alpha 1(COL1A1).The independent samples t-test was used for comparison of normally distributed continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups.Results Compared with normal liver tissue,the liver tissue of patients with hepatic fibrosis had significantly higher expression levels of PERK,eIF2α,and CHOP(Z=-3.56,t=-5.75,Z=-3.52,all P<0.001).Compared with the solvent group,the groups treated with different concentrations of thapsigargin had significant increases in the expression levels of the endoplasmic reticulum-associated proteins PERK,CHOP,IRE1,ATF6,and α-SMA(all P<0.05).Compared with the control group,the PERK stable overexpression group had significant increases in the mRNA expression levels of PERK,eIF2α,and α-SMA and the protein expression levels of PERK,p-eIF2α,and α-SMA(all P<0.05),and immunofluorescence assay showed a significant increase in the expression level of COL1A1 in the PERK stable overexpression group(P<0.05).Conclusion PERK overexpression can induce the expression of α-SMA and COL1A1 in LX-2 cells,suggesting that the PERK signaling pathway might be one of the important mechanisms of HSC activation.

Objective: To explore the effects of different concentrations of glucose solution on the survival of Aedes albopictus and Culex pipiens pallens larvae, the attraction to mosquitoes and egg-laying behaviors, so as to provide the reference for developing mosquito control technology based on sugar bait. Methods: White porcelain bowls were filled with 100 mL of 3%, 5%, 8%, 10% and 15% glucose solutions. Ten of fourth instar larvae of Aedes albopictus or Culex pipiens pallens were added to each bowl, and the survival of larvae was recorded after 2, 4, 6, 24, 48 and 72 hours. Egg-laying cups containing 5%, 8% and 15% glucose solution were put in mosquito cages containing fully blooded female mosquitoes of Aedes albopictus and Culex pipiens pallens (50 mosquitoes each), and the total number of eggs laid in 72 hours was observed. The analogous site room was filled with fully blooded and starved female mosquitoes of Aedes albopictus and Culex pipiens pallens (100 mosquitoes each), and simple mosquito control buckets containing 5% and 8% glucose solution and black sticky insect plates. The number of mosquitoes and eggs was observed after 6 days. All the above experiments were repeated 3 times using dechlorinated water as the control. Results: The 72 hour corrected mortality rates of Aedes albopictus and Culex pipiens pallens larvae gradually increased with the increase of glucose concentration. The glucose solution with 5% and higher concentrations was not suitable for mosquito larvae to survive. The attraction of egg-laying behaviors to Aedes albopictus and Culex pipiens pallens gradually decreased with the increase of glucose concentration. The effects were similar between 5% and 8% glucose solution, with the averages of 686.67 and 682.33 eggs for Aedes albopictus, and 3.00 and 2.33 egg rafts for Culex pipiens pallens. In analogous site room, there were 93.33, 105.00 and 130.33 adult mosquitoes captured on average in the control group, 5% and 8% glucose solution groups, respectively, with 8% glucose solution group more attractive to adult mosquitoes than the control group (F=3.283, P=0.030); there were 70.33, 55.33 and 63.00 Aedes albopictus eggs (eggs counts+larvae counts) on average, respectively, with statistically significant differences among the three groups (H=6.761, P=0.034). Conclusion Glucose solution with concentration of 5% or higher can effectively inhibit the survival of Aedes albopictus and Culex pipiens pallens larvae, and attractive to adult mosquitoes and egg-laying behavoirs.

The theory of “diagnosing diseases with sinews” means that through the diagnosis and examination of the channel sinews at the site of the lesion， the surplus and deficit state of qi and blood in the channel sinews， vessles and channels， and the degree of damage to the organism caused by the disease and evils， can be determined， forming the three elements （the nature of the disease， the location of the disease， and the disease tendency） of the disease diagnosis can be closely integrated to form the trinity of diagnostic modes， which is “examining the disease nature by sinews， identifying the pattern by sinews， and determining the tendency by sinews”. For intractable facial paralysis， the method of “diagnosing diseases with sinews” can be adopted， in which the morphological changes of the channel sinews are judged through diagnosis by observation， the traditional Chinese patterns are identified through diagnosis by palpation， and the points of meridian tendons and the circulation of tendon and treatment lines are determined through diagnosis by circulation. The “diagnosing diseases with sinews” not only helps to accurately determine the disease condition， patterns and development trend， but also helps to adopt targeted treatment for the disease and prevent the disease from spreading， and providing ideas and methods for the clinical diagnosis and treatment of intractable facial paralysis.

Objective:To explore the correlation between myocardial damage and vascular endothelial growth factor (VEGF), red blood cell distribution width (RDW), and myocardial enzyme spectrum in children with severe mycoplasma pneumoniae pneumonia.Methods:Sixty children with severe mycoplasma pneumoniae pneumonia and myocardial damage admitted to Jining Medical University from January 2019 to December 2020 were selected as the observation group, and 60 children with severe mycoplasma pneumoniae pneumonia admitted during the same period were selected as the control group. The differences in clinical data and imaging features between the two groups were compared. Pearson correlation analysis was used for correlation analysis; The logistic regression method was applied to analyze the influencing factors of myocardial damage in children with severe mycoplasma pneumoniae pneumonia. The value of VEGF and RDW in predicting myocardial damage in children with severe mycoplasma pneumoniae pneumonia was analyzed using receiver operating characteristic (ROC) curves.Results:The levels of C-reactive protein (CRP), procalcitonin (PCT), VEGF, RDW, creatine kinase isoenzyme (CK-MB), creatine kinase (CK), cardiac troponin I (cTnI), and lactate dehydrogenase (LDH) in the observation group were significantly higher than those in the control group (all P<0.05), and the duration of fever, application of macrolide drugs, and glucocorticoid application time were significantly longer than those in the control group (all P<0.05). There was no statistically significant difference in pulmonary imaging characteristics between the observation group and the control group (all P>0.05). The VEGF and RDW in the observation group were positively correlated with CK-MB and cTnI (all P<0.05). Logistic regression analysis showed that duration of fever, VEGF, RDW, and duration of macrolide drug use were the influencing factors for myocardial damage in children with severe mycoplasma pneumoniae pneumonia (all P<0.05). The area under the ROC curve of VEGF combined with RDW in predicting myocardial damage in children with severe mycoplasma pneumoniae pneumonia was 0.899, significantly higher than that predicted by VEGF and RDW alone (all P<0.05). Conclusions:The serum VEGF and RDW levels in children with severe mycoplasma pneumoniae pneumonia accompanied by myocardial damage are elevated and positively correlated with myocardial enzyme spectrum indicators, which has certain application value in predicting myocardial damage.

BACKGROUND:The implant osseointegration rate of patients with diabetes is low,and the failure rate is high,which seriously affects the quality of life.It is urgent to improve the implant osseointegration of patients with diabetes by effective means to elevate the success rate.Exploring the effect of berberine on the osteogenic differentiation of bone marrow mesenchymal stem cells under a high-glucose environment and its specific mechanism will provide effective theoretical support for solving the above problems. OBJECTIVE:To explore the effect of natural extract berberine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells under the high-glucose microenvironment. METHODS:Bone marrow mesenchymal stem cells of SD rats were cultured by the whole bone marrow adherence method.CCK-8 assay was used to detect the effects of different concentrations of berberine on the proliferation of bone marrow mesenchymal stem cells under the high-glucose environment and to screen out the optimal berberine concentration.The expressions of Runx2 and Osx were detected by alkaline phosphatase activity,alicarin red staining and PCR to determine the effect of berberine on osteogenic differentiation of bone marrow mesymal stem cells under the high-glucose environment.To further explore the underlying mechanism,we introduced the AMPK-specific inhibitor Dorsomorphin and used a DCFH-DA reactive oxygen species fluorescent probe to examine reactive oxygen species levels.The p-AMPK expression was also determined by western blot assay. RESULTS AND CONCLUSION:(1)10 μmol/L was the optimal concentration of berberine to promote bone marrow mesenchymal stem cell proliferation.(2)Alberberine promoted alkaline phosphatase viability of bone marrow mesenchymal stem cells and mineralized nodule formation in a high-glucose microenvironment.(3)Alberberine promoted the expression of Runx2 and OSx in a high-glucose microenvironment.(4)Alberensine effectively inhibited the reactive oxygen species level of bone marrow mesenchymal stem cells in a high-glucose environment.(5)The effects of berberine on promoting bone marrow mesenchymal stem cell osteogenesis and inhibition of reactive oxygen species were reversed by the AMPK inhibitor.(6)Berberine activated AMPK and promoted p-AMPK expression.(7)The above results indicate that berberine(10 μmol/L)promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment by activating AMPK and reducing intracellular reactive oxygen species levels.