The circadian clock plays a critical role in regulating various physiological processes, including gene expression, metabolic regulation, immune response, and the sleep-wake cycle in living organisms. Post-translational modifications (PTMs) are crucial regulatory mechanisms to maintain the precise oscillation of the circadian clock. By modulating the stability, activity, cell localization and protein-protein interactions of core clock proteins, PTMs enable these proteins to respond dynamically to environmental and intracellular changes, thereby sustaining the periodic oscillations of the circadian clock. Different types of PTMs exert their effects through distincting molecular mechanisms, collectively ensuring the proper function of the circadian system. This review systematically summarized several major types of PTMs, including phosphorylation, acetylation, ubiquitination, SUMOylation and oxidative modification, and overviewed their roles in regulating the core clock proteins and the associated pathways, with the goals of providing a theoretical foundation for the deeper understanding of clock mechanisms and the treatment of diseases associated with circadian disruption.
Protein Processing, Post-Translational/physiology* ; Circadian Rhythm/physiology* ; Humans ; Animals ; CLOCK Proteins/physiology* ; Circadian Clocks/physiology* ; Phosphorylation ; Acetylation ; Ubiquitination ; Sumoylation

This study aims to explore the protective effect of Chaihu Jia Longgu Muli Decoction on rats with heart failure after myocardial infarction, and to clarify its possible mechanisms, providing a new basis for basic research on the mechanism of classic Chinese medicinal formula-mediated inflammatory response in preventing and treating heart failure induced by apoptosis after myocardial infarction. A heart failure model after myocardial infarction was established in rats by coronary artery ligation. The rats were divided into sham group, model group, and low, medium, and high-dose groups of Chaihu Jia Longgu Muli Decoction, with 10 rats in each group. The low-dose, medium-dose, and high-dose groups of Chaihu Jia Longgu Muli Decoction were given 6.3, 12.6, and 25.2 g·kg~(-1) doses by gavage, respectively. The sham group and model group were given an equal volume of distilled water by gavage once daily for four consecutive weeks. Cardiac function was assessed using color Doppler echocardiography. Myocardial pathology was detected by hematoxylin-eosin(HE) staining, apoptosis was measured by TUNEL assay, and mitophagy was observed by transmission electron microscopy. The levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, and N-terminal pro-B-type natriuretic peptide(NT-proBNP) in serum were detected by enzyme-linked immunosorbent assay(ELISA). The expression of apoptosis-related proteins B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), and cleaved caspase-3 was detected by Western blot. Additionally, the expression of phosphorylated nuclear transcription factor-κB(NF-κB) p65(p-NF-κB p65)(upstream) and nuclear factor kappa B inhibitor alpha(IκBα)(downstream) in the NF-κB signaling pathway was assessed by Western blot. The results showed that compared with the sham group, left ventricular ejection fraction(LVEF) and left ventricular short axis shortening(LVFS) in the model group were significantly reduced, while left ventricular end diastolic diameter(LVEDD) and left ventricular end systolic diameter(LVESD) increased significantly. Myocardial tissue damage was severe, with widened intercellular spaces and disorganized cell arrangement. The apoptosis rate was increased, and mitochondria were enlarged with increased vacuoles. Levels of TNF-α, IL-1β, and NT-proBNP were elevated, indicating an obvious inflammatory response. The expression of pro-apoptotic factors Bax and cleaved caspase-3 increased, while the anti-apoptotic factor Bcl-2 decreased. The expression of p-NF-κB p65 was upregulated, and the expression of IκBα was downregulated. In contrast, the Chaihu Jia Longgu Muli Decoction groups showed significantly improved of LVEF, LVFS and decreased LVEDD, LVESD compared to the model group. Myocardial tissue damage was alleviated, and intercellular spaces were reduced. The apoptosis rate decreased, mitochondrial volume decreased, and the levels of TNF-α, IL-1β, and NT-proBNP were lower. The expression of pro-apoptotic factors Bax and cleaved caspase-3 decreased, while the expression of the anti-apoptotic factor Bcl-2 increased. Additionally, the expression of p-NF-κB p65 decreased, while IκBα expression increased. In summary, this experimental study shows that Chaihu Jia Longgu Muli Decoction can reduce the inflammatory response and apoptosis rate in rats with heart failure after myocardial infarction, which may be related to the regulation of the IκBα/NF-κB signaling pathway.
Animals ; Apoptosis/drug effects* ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Myocardial Infarction/physiopathology* ; Male ; NF-kappa B/genetics* ; Heart Failure/etiology* ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; NF-KappaB Inhibitor alpha/genetics* ; Humans ; Tumor Necrosis Factor-alpha/genetics*

A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL^-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.

In recent years,the prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection has become a global public health issue.Ceftazidime/avibactam(CAZ/AVI)has been approved as a novel antimicrobial agent for the treatment of healthcare-associated pneumonia/ventilator-associated pneumonia,bloodstream infection,infection after kidney transplantation,and severe infection combined with liver cirrhosis.However,the use of CAZ/AVI has also led to the emergence of drug-resistant strains.The major mechanisms of drug-resistance include over-expression of blaKPC gene,mutation of β-lactamase and amino acids at key sites,changes in cell permeability caused by loss of membrane porin,and over-expression of efflux pump.This article reviews the research progress of CAZ/AVI in the treatment of CRKP infection,providing reference for clinical diagnosis and treatment.

Aim To investigate the effect of discoidin domain receptor 1(DDR1)on high glucose induced endothelial cell dysfunction and the underlying mecha-nism.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided in-to the control group and high glucose induction group(HG).HUVECs were treated with 33 mmol·L-1 D-glucose for 48 hours to construct endothelial dysfunc-tion.Pyroptosis was detected using propidium iodide staining(PI);lactate dehydrogenase(LDH)and IL-1β,IL-18 levels were determined using enzyme linked immunosorbent assay(ELISA);the expression of DDR1 and NF-κB/NLRP3 signaling pathway proteins and pyroptosis related proteinses were detected using Western blot.Subsequently,the experiment was divid-ed into the control group,HG group,HG+DDR1 NC group,and HG+DDR1 siRNA group.The effect of high glucose on the proliferation and migration of HU-VECs was observed after transfection with DDR1 siR-NA for 24 hours;ELISA was used to detect the endo-thelial nitric oxide synthase(eNOS),vascular cell ad-hesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),as well as LDH,IL-1β,IL-18 levels;PI was employed to detect pyroptosis;Western blot was applied to detect DDR1 and NF-κB/NLRP3 signaling pathway proteins and pyroptosis related pro-teins.Results Compared with the control group,HG group decreased eNOS content,increased VCAM-1 and ICAM-1 contents,decreased cell viability and migration ability,and significantly increased the expressions of DDR1,p-NF-κB,NLRP3 and pyroptosis related pro-teins.The levels of LDH,IL-1β,IL-18 and the rate of pyroptosis significantly increased(P＜0.05).Com-pared with HG group,DDR1 siRNA could promote the secretion of eNOS,decrease the levels of VCAM-1,ICAM-1,LDH,IL-1β and IL-1 8,increase cell viability and migration ability,reduce the expression of p-NF-κB,NLRP3 and pyroptosis related proteins,and inhibit high glucose-induced pyroptosis of HUVECs(P＜0.05).Conclusions Gene silencing DDR1 can im-prove vascular endothelial cell dysfunction induced by high glucose,and the mechanism is related to the inhi-bition of NF-κB/NLRP3 signaling pathway mediated pyroptosis.

Objective: To explore the allergen map of patients with allergic diseases in northwest China, to investigate the distribution characteristics of serum specific Immunoglobulin E (sIgE) in different ages, genders, diseases and the sensitization patterns of allergens. Methods: This study is a cross-sectional study, a total of 1 161 patients with confirmed respiratory allergic diseases were selected retrospectively from outpatient or inpatient department of Gansu Provincial People's Hospital, Gansu Provincial Maternity and Child Care Hospital, General Hospital of Ningxia Medical University, Yinchuan Maternal and Child Health Care Hospital, Xijing Hospital, Air Force Military Medical University and Tumor Hospital of Inner Mongolia Autonomous Region from June 2019 to October 2022. HAIKE ALLEOS 2000 fluorescent magnetic particle chemiluminescence method was used to quantify their serum for 12 inhaled allergen-specific IgE. Chi square test or Fisher's exact test were used for comparison between count data groups (Bonferroni correction was used for further pairwise comparison in multiple groups, two-sided P<0.05/3=0.017 considered that the difference was statistically significant). Pearson correlation analysis was used for correlation of continuous numerical variables. Results: The positive detection rate of sIgE in 1 161 patients was 66.8%(776/1 161). The three highest positive rate of inhaled allergen were mugwort(599/1 161, 51.6%), French chrysanthemum(565/1 161, 48.7%) and dandelion(412/1 161, 35.5%). In different age groups, the highest positive rate of sIgE was 7-18 age group(379/513, 73.9%) while the lowest positive rate was 4-6 age group(222/370, 60.0%), the difference between groups was statistically significant(χ²=21.177, P<0.001). The sensitization peak of mugwort, French chrysanthemum, plantain, timothy, birch, dermatophagoides pteronyssinus, dermatophagoides farinae, cat epithelium, dog epithelium and German cockroach appeared in 7-18 age group. In different disease groups, the highest positive rate of sIgE was allergic rhinitis with asthma group (500/717, 69.7%) while the lowest positive rate was asthma group (76/144, 52.8%), the difference between groups was statistically significant(χ²=15.563, P<0.001). In different gender groups, the positive rate of sIgE in male (503/711, 70.7%) was higher than in female (273/450, 60.7%), the difference between groups was statistically significant(χ²=12.630, P<0.001). The multiple-sensitization rate was 86.9%(674/776) and the double-sensitization rate was 16.8%(130/776) in sIgE positive patients. Pearson correlation results showed that there was an extremely strong correlation between dandelion and French chrysanthemum(r=0.93,P<0.001). There was a strong correlation between mugwort and French chrysanthemum(r=0.64,P<0.001). In the co-sensitization analysis, the number of patients sensitized by mugwort, French chrysanthemum, dandelion, plantain and timothy accounted for 25.2%(170/674)of the total number of multiple sensitization. The number of patients sensitized by mugwort, French chrysanthemum and dandelion accounted for 58.3%(393/674)of the total number of multiple sensitization. The number of patients sensitized by mugwort, French chrysanthemum, dandelion and plantain accounted for 35.6%(240/674) of the total number of multiple sensitization. Conclusion: Mugwort, French chrysanthemum, dandelion are the major inhaled allergens in northwest China. The positive rate of sIgE was different in different ages, diseases and genders. The multiple-sensitization rate of allergen was high and there was a certain positive correlation between pollen allergen-specific IgE pairwise, suggesting that there may exist co-sensitization or cross-reactions among allergens.
Pregnancy ; Child ; Animals ; Dogs ; Female ; Humans ; Male ; Allergens ; Cross-Sectional Studies ; Retrospective Studies ; Respiratory System ; Asthma

Objective: To explore the allergen map of patients with allergic diseases in northwest China, to investigate the distribution characteristics of serum specific Immunoglobulin E (sIgE) in different ages, genders, diseases and the sensitization patterns of allergens. Methods: This study is a cross-sectional study, a total of 1 161 patients with confirmed respiratory allergic diseases were selected retrospectively from outpatient or inpatient department of Gansu Provincial People's Hospital, Gansu Provincial Maternity and Child Care Hospital, General Hospital of Ningxia Medical University, Yinchuan Maternal and Child Health Care Hospital, Xijing Hospital, Air Force Military Medical University and Tumor Hospital of Inner Mongolia Autonomous Region from June 2019 to October 2022. HAIKE ALLEOS 2000 fluorescent magnetic particle chemiluminescence method was used to quantify their serum for 12 inhaled allergen-specific IgE. Chi square test or Fisher's exact test were used for comparison between count data groups (Bonferroni correction was used for further pairwise comparison in multiple groups, two-sided P<0.05/3=0.017 considered that the difference was statistically significant). Pearson correlation analysis was used for correlation of continuous numerical variables. Results: The positive detection rate of sIgE in 1 161 patients was 66.8%(776/1 161). The three highest positive rate of inhaled allergen were mugwort(599/1 161, 51.6%), French chrysanthemum(565/1 161, 48.7%) and dandelion(412/1 161, 35.5%). In different age groups, the highest positive rate of sIgE was 7-18 age group(379/513, 73.9%) while the lowest positive rate was 4-6 age group(222/370, 60.0%), the difference between groups was statistically significant(χ²=21.177, P<0.001). The sensitization peak of mugwort, French chrysanthemum, plantain, timothy, birch, dermatophagoides pteronyssinus, dermatophagoides farinae, cat epithelium, dog epithelium and German cockroach appeared in 7-18 age group. In different disease groups, the highest positive rate of sIgE was allergic rhinitis with asthma group (500/717, 69.7%) while the lowest positive rate was asthma group (76/144, 52.8%), the difference between groups was statistically significant(χ²=15.563, P<0.001). In different gender groups, the positive rate of sIgE in male (503/711, 70.7%) was higher than in female (273/450, 60.7%), the difference between groups was statistically significant(χ²=12.630, P<0.001). The multiple-sensitization rate was 86.9%(674/776) and the double-sensitization rate was 16.8%(130/776) in sIgE positive patients. Pearson correlation results showed that there was an extremely strong correlation between dandelion and French chrysanthemum(r=0.93,P<0.001). There was a strong correlation between mugwort and French chrysanthemum(r=0.64,P<0.001). In the co-sensitization analysis, the number of patients sensitized by mugwort, French chrysanthemum, dandelion, plantain and timothy accounted for 25.2%(170/674)of the total number of multiple sensitization. The number of patients sensitized by mugwort, French chrysanthemum and dandelion accounted for 58.3%(393/674)of the total number of multiple sensitization. The number of patients sensitized by mugwort, French chrysanthemum, dandelion and plantain accounted for 35.6%(240/674) of the total number of multiple sensitization. Conclusion: Mugwort, French chrysanthemum, dandelion are the major inhaled allergens in northwest China. The positive rate of sIgE was different in different ages, diseases and genders. The multiple-sensitization rate of allergen was high and there was a certain positive correlation between pollen allergen-specific IgE pairwise, suggesting that there may exist co-sensitization or cross-reactions among allergens.
Pregnancy ; Child ; Animals ; Dogs ; Female ; Humans ; Male ; Allergens ; Cross-Sectional Studies ; Retrospective Studies ; Respiratory System ; Asthma

Objective: To investigate the clinical and molecular biological characteristics of patients with accelerated chronic lymphocytic leukemia (aCLL) . Methods: From January 2020 to October 2022, the data of 13 patients diagnosed with aCLL at The First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed to explore the clinical and molecular biological characteristics of aCLL. Results: The median age of the patients was 54 (35-72) years. Prior to aCLL, five patients received no treatment for CLL/small lymphocytic lymphoma (SLL), while the other patients received treatment, predominantly with BTK inhibitors. The patients were diagnosed with aCLL through pathological confirmation upon disease progression. Six patients exhibited bulky disease (lesions with a maximum diameter ≥5 cm). Positron emission tomography (PET) -computed tomography (CT) images revealed metabolic heterogeneity, both between and within lesions, and the median maximum standardized uptake value (SUVmax) of the lesion with the most elevated metabolic activity was 6.96 (2.51-11.90). Patients with unmutated IGHV CLL accounted for 76.9% (10/13), and the most frequent genetic and molecular aberrations included +12 [3/7 (42.9% ) ], ATM mutation [6/12 (50% ) ], and NOTCH1 mutation [6/12 (50% ) ]. Twelve patients received subsequent treatment. The overall response rate was 91.7%, and the complete response rate was 58.3%. Five patients experienced disease progression, among which two patients developed Richter transformation. Patients with aCLL with KRAS mutation had worse progression-free survival (7.0 month vs 26.3 months, P=0.015) . Conclusion: Patients with aCLL exhibited a clinically aggressive course, often accompanied by unfavorable prognostic factors, including unmutated IGHV, +12, ATM mutation, and NOTCH1 mutation. Patients with CLL/SLL with clinical suspicion of disease progression, especially those with bulky disease and PET-CT SUVmax ≥5, should undergo biopsy at the site of highest metabolic uptake to establish a definitive pathological diagnosis.
Humans ; Middle Aged ; Aged ; Leukemia, Lymphocytic, Chronic, B-Cell/genetics* ; Positron Emission Tomography Computed Tomography ; Retrospective Studies ; Biopsy ; Disease Progression

A retrospective study was conducted on data of 23 patients with pancreatic duct diseases who were underwent peroral pancreatoscopy (POPS) at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from August 2020 to October 2022. The intraoperative observation, postoperative complications, and the diagnosis and treatment of POPS for pancreatic duct diseases were analyzed. All patients underwent POPS and achieved technical success. Among them, 7 patients were diagnosed as having intraductal papillary mucinous neoplasm of pancreas and 3 pancreatic malignant tumor. Eight patients with pancreatolithiasis accepted laser or eletrohydraulic lithotripsy under POPS. Abdominal pain improved in 2 patients with chronic pancreatitis after treatment. Melena disappeared in 2 patients with pancreatic duct hemorrhage or pancreatic enterostomy inflammation after conservative treatment. The symptom of 1 patient with pancreatic enterostomy stenosis improved after balloon dilation. There was no complication in the 23 patients, and the operation time was 35-90 min. The results indicate POPS is safe, effective with distinctive advantages in the diagnosis and treatment for pancreatic duct diseases.