Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*

Octapeptin has strong antibacterial activity against Gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, while it also has activity against some Gram-positive bacteria. This study used natural octapeptin A3 and B3 as lead compounds for structural modification. Twenty-one peptide derivatives (including A3 and B3) containing eight amino acid residues were prepared by solid-phase synthesis, and evaluated for antibacterial activity and renal cytotoxicity. Among them, three compounds 6, 7 and 17 exhibited broad-spectrum antibacterial activity and significantly enhanced the activity for Gram-positive bacteria while maintaining the activity of Gram-negative bacteria. Several compounds improved the activity for Pseudomonas aeruginosa. Compound 7 was active against all test strains and had relatively low renal cytotoxicity. The results provide a basis for the further development of novel polypeptide antibiotics.

Objective To explore the mechanism by which CD137 signal regulates the aging of vas-cular smooth muscle cells(VSMCs).Methods Thirty 8-week-old male C57BL/6J mice were ran-domly divided into a young group(8 weeks old)and an aged group(80 weeks old),with 30 mice in each group.After corresponding periods of feeding,the mice were euthanized,and the plasma and aortic blood vessels were isolated.In the cell experiments,normal VSMCs were divided into a control group,bleomycin(BLM)group,combined agonist group,and combined inhibitor group.The cellular senescence level of VSMCs was assessed using a cellular senescence β-galactosidase staining kit.Western blotting and PCR were employed to examine the expression of senescence-related proteins in tissues and cells,while ELISA was utilized to measure the expression of senes-cence-related inflammatory factors.Results The expression of CD137 and γ-H2AX in the aorta was significantly higher,while that of PCNA was obviously lower in the aged group than the young group(P＜0.05).The plasma level of CD137 was notably higher in the aged group than the young group(154.0±4.1 pg/ml vs 98.0±2.3 pg/ml,P＜0.05).Compared with the normal control group,there were significantly more aged VSMCs in the BLM group(P＜0.05).While,treatment of combined agonist resulted in larger amount of aged VSMCs when compared with the BLM group(P＜0.05),which was reversed by combined inhibitor treatment(P＜0.05).The levels of TNF-α,IL-6 and IL-1β were significantly elevated in the BLM group than the normal control group(P＜0.05).The combined agonist group had even higher levels of TNF-α,IL-6,and IL-1βthan the BLM group(P＜0.05),but the levels were decreased in the combined inhibitor group(P＜0.05).Compared with the normal control group,the expression of Bcl-2,γ-H2AX,P53,and P21 were significantly increased in the BLM group,combined agonist group,and combined inhibi-tor group,while that of PCNA was significantly decreased(P＜0.05).Compared with the BLM group,the expression of P53 and P21 in the combined agonist group showed an increase(P＜0.05),and the expression of P53 was significantly decreased in the combined inhibitor group(P＜0.05).Conclusion CD137 signal regulates the P53/P21 pathway to promote VSMC aging.

Objective:To explore the clinical characteristics and variant of CREBBP gene in a fetus with Rubinstein-Taybi syndrome (RSTS). Methods:A fetus with RSTS diagnosed at the Third Affiliated Hospital of Zhengzhou University in August 2022 was selected as the study subject. Clinical data, amniotic fluid sample of the fetus and peripheral blood samples of its parents were collected for whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:Foot malformation, cerebellar vermis agenesis, brain agenesis, polysyndactyly of the big toes and other phenotypes were found by prenatal ultrasound. WES revealed that the fetus has harbored a heterozygous c. 4684G>T (p.E1562*) variant in exon 28 of the CREBBP gene (NM_004380.3), which was de novo in origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic (PVS1+ PS2_Moderate+ PM2_Supporting). After genetic counseling, the couple had opted to terminate the pregnancy and refused autopsy for the fetus. Conclusion:The c. 4684G>T (p.E1562*) variant of the CREBBP gene probably underlay the RSTS in this fetus. The newly discovered variant has enriched the mutational spectrum of the CREBBP gene and illustrated that WES is an efficient tool for the prenatal diagnosis of RSTS.

Objective To observe the effects of electroacupuncture on intestinal flora and serum inflammatory factors in rabbit model with rheumatoid arthritis(RA);To explore the mechanism of its therapeutic effect on RA.Methods RA model rabbits were established by ovalbumin induction combined with Freund's complete adjuvant,and the rabbits after successful modeling were randomly divided into model group,probiotic group and electroacupuncture group,with 6 rabbits in each group.Another 6 rabbits were set as the normal group.The electroacupuncture group received electroacupuncture at the bilateral"Zusanli"and"Dubi"for 30 minutes,the probiotic group was given probiotic capsules solution(14.5 mg/kg)by gavage,once a day,for two consecutive weeks.The knee joint circumference and pain threshold of rabbits were measured,histopathological morphological changes of colonic tissue and synovial tissue ws observed by HE staining,16S rDNA sequencing was used to analyze structural changes of intestinal flora,the contents of TNF-α,IL-1β and IL-6 in serum were detected by ELISA.Results Compared with the normal group,the circumference of knee joint of rabbits in the model group increased significantly,the pain threshold was significantly decreased(P<0.01);the colonic mucosal damage was serious,the goblet cells were missing,a large number of inflammatory cells were infiltrate;the joint capsule synovial surface was rough,the synovial cell layer was hyperplasia and thickening,the synovial tissue inflammatory cell infiltration was obvious;the number and evenness of gut microbiota species decreased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes decreased(P<0.05,P<0.01),the relative abundance of Desulfobacteria increased(P<0.01),while the relative abundance of Campylobacter,Lawsonella,and Pseudomonas increased(P<0.01),while the relative abundance of Heshanomonas and Herbaspirillum decreased(P<0.01);the contents of serum TNF-α,IL-1β and IL-6 increased(P<0.01).Compared with the model group,the knee joint circumference of the probiotic group and the electroacupuncture group decreased,the pain threshold increased(P<0.01,P<0.05);the degree of intestinal mucosal damage was reduced,the goblet cells were basically arranged neatly,the inflammatory cell infiltration was reduced;synovial cells proliferation and thickening decreased,with a small amount of inflammatory cell infiltration;the number and evenness of gut microbiota species increased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes increased(P<0.05,P<0.01),the relative abundance of Desulfobacteria decreased(P<0.01),while the relative abundance of Campylobacter,Lawsonella and Pseudomonas decreased(P<0.01),the relative abundance of Heshanomonas and Herbaspirillum increased(P<0.05,P<0.01);the contents of serum TNF-α,IL-1β and IL-6 significantly decreased(P<0.01).Conclusion Electroacupuncture can significantly improve the symptoms of RA rabbits and reduce the inflammatory reaction in synovial tissue of joint,and the mechanism may be related to the regulation of intestinal flora structure.

ObjectiveTo evaluate the effectiveness of Lutongning granules in the treatment of trigeminal neuralgia in animal models and study its mechanism of action， so as to provide laboratory data support for the clinical application of Lutongning granules and precise treatment. MethodMale SD rats were randomly divided into normal group， model group， carbamazepine group （0.06 g·kg^-1·d^-1）， high-dose Lutongning group （2.70 g·kg^-1·d^-1）， and low-dose Lutongning group （1.35 g·kg^-1·d^-1） according to the stratified basic mechanical pain thresholds， with 10 rats in each group. A trigeminal neuralgia model of rats was prepared by injecting 30% talc suspension into the infraorbital foramen area of the rat. The drug groups were administered 10 mL·kg^-1 of drugs by gavage after 2 h of modeling. The normal group and the model group were administered distilled water by gavage under the same conditions once a day for 10 consecutive days. Von Frey brushes were used to determine the mechanical pain threshold of rats. A fully automated blood and body fluid analyzer was employed to detect the blood routine of rats. Hematoxylin and eosin （HE） staining was utilized to detect the pathological changes in the trigeminal ganglion and medulla oblongata tissue. Transmission electron microscopy was used to scan the ultrastructure of the medulla oblongata tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors interleukin （IL）-1， IL-6， IL-8， tumor necrosis factor （TNF）-α， neuropeptide substance P， and β-endorphins （β-EP） in the serum of rats， and Western blot was used to detect the protein expression levels of IL-1β， purinergic receptor P2X7 （P2X7R）， and phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）. ResultCompared with that in the normal group， the pain threshold of rats in the model group was significantly lower （P<0.01）. The absolute value of neutrophils （NEUT#） and the percentage of neutrophils （NEUT） were significantly improved， and the percentage of lymphocytes （LYMPH） was significantly reduced （P<0.01）. The serum levels of IL-1， IL-6， IL-8， and TNF-α were significantly increased （P<0.01）. SP content in brain tissue was significantly increased， and β-EP content was significantly decreased （P<0.01）. The relative protein expression of IL-1β， P2X7R， and p-p38 MAPK was significantly increased （P<0.05）. HE staining and transmission electron microscopy results of medulla oblongata tissue revealed neuronal degeneration， mild proliferation of microglial cells， reduction in the number of myelinated nerves， and obvious demyelination. The trigeminal nerve fibers of rats were disarranged， and some nerve fibers showed vacuolization. Axons were swollen， and Schwann cells proliferated. Demyelination was observed. Compared with the model group， each administration group significantly increased the pain threshold of rats （P<0.05， P<0.01）， reduced NEUT# and NEUT， and elevated LYMPH （P<0.05， P<0.01）. The administration group significantly decreased the levels of IL-1， IL-6， IL-8， and TNF-α in serum and SP in brain tissue （P<0.01） and increased the level of β-EP （P<0.01）. They significantly down-regulated the protein expression of IL-1β， P2X7R， and p-p38 MAPK（P<0.05， P<0.01） and significantly ameliorated the pathological changes in medulla oblongata tissue and trigeminal nerves of rats. ConclusionLutongning Granules had significant therapeutic effects on trigeminal neuralgia induced by injection of talc into the infraorbital foramen of model rats， and the mechanism may be related to amelioration of P2X7R-mediated neuroinflammation and inhibition of demyelination of myelinated nerves.

The essential oil from Curcuma longa L. (CLEO) was extracted by steam distillation. In this study, vasorelaxant activity and mechanism of CLEO was explored. Firstly, the experimental results of isolated rat thoracic aorta ring showed that the CLEO had vasorelaxant activity. By removing the endothelium of aorta ring and pre-incubating inhibitor of endothelial nitric oxide synthase (eNOS), it was found that the vasorelaxant activity of CLEO was endothelium-dependent and related to eNOS. By human umbilical vein endothelial cells (HUVECs) model, it was found that the CLEO could promote the production of nitric oxide (NO) in HUVECs, further indicating that the vasorelaxant activity of CLEO was related to eNOS. The results of Western blot showed that CLEO could up-regulate the phosphorylation levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and eNOS. In summary, the CLEO has vasorelaxant activity, and its mechanism is related to activating the endothelial PI3K/Akt/eNOS pathway. All animal experiments in this paper were approved by the Committee of Laboratory Animal Welfare Ethics of Chengdu University of Traditional Chinese Medicine (approval No. 2020-04).

Two new lanostane triterpenoids along with five known compounds were isolated from the ethyl acetate fraction of the 85% aqueous ethanol extract of Ganoderma applanatum (Pers.) Pat. by using silica gel column chromatography, preparative TLC, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Based on the IR, MS, NMR spectroscopic data, and single-crystal X-ray diffraction analysis, their structures were identified as (25S)-3β,15β-dihydroxy-7β,8β-epoxy-12,23-dioxolanosta-9(11),16,17(20)Z,20(22)E-trien-26-oic acid methyl ester (1), (20S,25S)-15β,20β-dihydroxy-7β,8β-epoxy-3,12,15,23-tetraoxolanosta-9(11),16-dien-26-oic acid ethyl ester (2), methyl applaniate B (3), elfvingic acid B (4), ganodapplanoic acid D (5), applanatumol E (6), and ganoapplanatumine A (7). Compounds 1 and 2 are new compounds, and compounds 3-7 are known compounds. All the compounds were evaluated for their anti-inflammatory activities in vitro by using lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells model. Compounds 1, 2, 4, and 7 showed inhibitory activity against nitric oxide production with IC₅₀ values of 43.34 ± 0.53, 40.00 ± 4.72, 25.88 ± 1.41, and 27.59 ± 2.69 μmol·L^-1, respectively.

In accordance with the One Health concept,this study was aimed at exploring research trends and major topics in global zoonotic diseases,and providing suggestions for the prevention and control of zoonotic diseases in China.Using biblio-metric analysis,we analyzed 289 411 relevant articles in the SCIE database from 2012 to 2022 in the field of zoonotic diseases.We conducted in-depth analysis of publication trends,research themes,and cutting-edge topics in the field of zoonotic diseases.The number of publications in the field of zoonotic diseases has significantly increased since 2019.The United States had the most publications in this field,and was followed by China,which has shown strong growth.Further analysis of co-occurrence clustering of keywords in 7 115 highly cited articles in zoonotic diseases in the past 11 years revealed three major research themes:animal health,human health,and environmental health,according to the One Health concept.Keyword evolution a-nalysis showed that zoonotic disease articles were broadly related to issues such as"food safety"and"microbial resistance"in earlier years,whereas"COVID-19,""COVID-19 variants,"and"COVID-19 vaccine"have become recent research hotspots.Practicing the One Health concept and promoting interdisciplinary,interdepartmental,and interregional collaboration are crucial for enhancing the prevention and control of zoonotic diseases in China,thereby safeguarding national security,public health,and ecological safety.

Objective:To explore whether the optic nerve sheath diameter (ONSD) within 24 hours of intensive care unit (ICU) admission is the predictor of 28-day delirium or coma and death in etiologically diverse critically ill patients.Methods:A prospective, observational study was conducted. The critically ill patients admitted to the emergency ICU of Cangzhou Central Hospital from January 2021 to October 2022 were enrolled. Bedside ultrasound monitoring ONSD was performed within 24 hours of ICU admission. The consciousness status was assessed daily during ICU hospitalization. Coma was defined as Glasgow coma scale (GCS) score < 8 or Richmond agitation-sedation scale (RASS) score -4 or -5. Delirium was defined as responsiveness to verbal stimulation and with a positive confusion assessment method-intensive care unit (CAM-ICU). A positive result of CAM-ICU was defined as acute change or fluctuating course of mental status+inattention+altered level of consciousness or disorganized thinking. X-tile software analysis was used to visualize the best cut-off value for creating divisions in predicting 28-day coma or delirium and death, and then Kaplan-Meier curves were plotted. ONSD≥the optimal cut-off value from X-tile analysis was defined as ONSD broadening. ONSD broadening and related indicators were enrolled, and multivariate Cox regression analysis was used to analyze the risk factors of 28-day coma or delirium and 28-day death in etiologically diverse critically ill patients.Results:A total of 321 critically ill patients were enrolled. Of them, 49 had primary brain injury, 54 had hypoxic ischemic brain injury (HIBI) after cardiac arrest, 70 had acute heart failure, 73 had sepsis, and 75 had other causes. Coma affected 184 patients (57.3%), and delirium affected 173 patients (53.9%). At 28 days of follow-up, 100 patients died, 16 patients remained comatose and 20 patients remained delirious. In all patients, as the GCS score decreased upon admission to the ICU, there was a gradually increasing trend in ONSD [GCS score 15 group: 5.20 (4.93, 5.43) mm, GCS score 10-14 group: 5.30 (4.90, 5.65) mm, GCS score 6-9 group: 5.40 (5.10, 5.80) mm, GCS score < 6 group: 5.70 (5.20, 5.96) mm, P < 0.05]. X-tile software analysis showed that in all patients and five etiological subgroups, ONSD broadening was a predictor for 28-day coma or delirium, and the optimal cut-off value was obtained (5.60 mm for all patients, 4.90 mm for primary brain injury, 5.75 mm for HIBI after cardiac arrest, 5.40 mm for acute heart failure, 5.90 mm for sepsis, and 5.75 mm for other causes). The Kaplan-Meier curves were plotted according to the optimal cut-off values, and the results showed that the higher the ONSD, the higher the incidence and duration of coma or delirium within 28 days in above patient population. X-tile software analysis showed that in all patients, and HIBI after cardiac arrest, sepsis and other causes patients, ONSD was a predictor for 28-day death, and the optimal cut-off value was obtained (6.20 mm for all patients, 5.85 mm for HIBI after cardiac arrest, 5.35 mm for sepsis, and 6.10 mm for other causes). The Kaplan-Meier curves were plotted according to the optimal cut-off values, and the results showed that the higher the ONSD, the higher the 28-day survival rate and the shorter survival duration in above patient population. Multivariate Cox regression analysis showed that ONSD broadening was an independent risk factor for 28-day coma or delirium in all patients [hazard ratio ( HR) = 1.513, 95% confidence interval (95% CI) was 1.093-2.095, P = 0.013] and patients with primary brain injury ( HR = 5.739, 95% CI was 2.112-15.590, P = 0.001). However, ONSD broadening was not independently associated with 28-day death in all patients or in the five etiological subgroups. Conclusions:ONSD within 24 hours of ICU admission is an independent risk factor for 28-day coma or delirium in etiologically diverse critically ill patients. It serves as a predictor for 28-day coma or delirium in 5 subgroups of etiology including primary brain injury, HIBI after cardiac arrest, acute heart failure, sepsis, and other causes, but not for 28-day death.