ObjectiveTo explore the mechanism by which the ethanol extract of Epimedium brevicornu （EEBM） intervenes in mesenchymal adipose-derived stem cells （ADSCs） to delay aging in castrated rats. MethodsForty-five 3-month-old SPF female SD rats were ovariectomized and randomly divided into model group， ADSCs treatment group， and ADSCs groups treated with low， medium， and high concentrations of EEBM （1， 50， 100 μg·L^-1）， referred to as the AE low， medium， and high concentration groups， with 9 rats in each group. After tail vein injection of 200 μL of the corresponding stem cell suspension， aging-related indicators including cyclin-dependent kinase inhibitor （p21）， tumor suppressor gene （p53）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， superoxide dismutase （SOD）， malondialdehyde （MDA）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cysteine-aspartic acid protease-3 （Caspase-3）， and lipofuscin were measured using enzyme-linked immunosorbent assay （ELISA） and Western blot. ResultsCompared with the model group， the IL-6 content in the AE low， medium， and high concentration groups was significantly decreased （P<0.05）. Lipofuscin， MDA， and IL-8 levels in the ADSCs treatment group and AE low， medium， and high concentration groups were significantly reduced （P<0.01）， while SOD content was significantly increased （P<0.05， P<0.01）. Compared with the ADSCs treatment group， lipofuscin and IL-8 levels in the AE low， medium， and high concentration groups were significantly reduced （P<0.05， P<0.01）. The MDA content was significantly decreased in the AE medium concentration group （P<0.01）. Compared with the model group， protein levels of p21， p53， Bax， and Caspase-3 in the ADSCs treatment group and AE low， medium， and high concentration groups were significantly reduced （P<0.05， P<0.01）， while the Bcl-2 protein level was significantly increased （P<0.01）. Compared with the ADSCs treatment group， protein levels of p21， p53， Bax， and Caspase-3 in the AE low， medium， and high concentration groups were significantly reduced （P<0.05， P<0.01）， and the Bcl-2 protein level in the AE low concentration group was significantly increased （P<0.01）. ConclusionThe results of this experiment show that EEBM-treated ADSCs or ADSCs may delay aging in castrated rats by inhibiting cell apoptosis， reducing cell cycle inhibitors and pro-inflammatory factors， enhancing antioxidant capacity， and reducing oxidative reactions. Moreover， EEBM-treated ADSCs demonstrate stronger anti-aging effects than ADSCs alone. This study provides experimental evidence supporting the clinical use of EEBM to intervene in ADSCs and delay aging.

Objective To study epigenetic diversity of Astragalus mongolicus by methylation-sensitive amplification polymorphism(MSAP)technique,and to establish and optimize the response system of MSAP technique applicable to Astragalus Mongolicus.Methods Astragalus membranaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao was used as material.The single factor and orthogonal design method were used to optimize the key influencing factors of the reaction system of Astragalus Mongolian methylation-sensitive amplification polymorphism(MSAP)technique,and the best reaction system of Astragalus Mongolian MSAP was established.Results 0.5 μL each of EcoR I and Msp I/Hpa II were added to the 20 μL double-enzyme digestion reaction system,and the digestion was completed in a water bath at 37℃for 6 h to complete the digestion;the optimal ligation system was 25 μL,including 10 μL digestion product,0.2 μL T4 DNA Ligase,1 μL EcoR I-adapter,1 μL Hpa II/Msp I-adapter,2.5 μL 10×T4 Buffer;25 μL optimal pre-amplification reaction system,including 2.5 μL 10×PCR buffer,0.5 μL dNTP,2.0 μL ligation product,0.3 μL Taq DNA polymerase,1.0 μL upstream and downstream primers;25 μL optimal selective amplification system,including 2 μL 20-fold diluted pre-amplification product,1.0 μL 10×PCR buffer,1.0 μL dNTP,0.2 μL Taq DNA polymerase,0.8 μL primers.Conclusion The optimized MSAP system was detected by polyacrylamide gel electrophoresis,which showed that the polymorphism was good,the system was stable,the bands were clear and repeatable,which laid the foundation for the subsequent MSAP analysis of Astragalus mongolica.

ObjectiveTo study the inhibitory effect of polyphyllin Ⅰ （PPI） on the growth of colorectal cancer cells and its molecular mechanism. MethodRKO cells were cultured and divided into a blank group and PPI treatment groups with concentrations of 0.6， 0.8， 1.0 μmol·L^-1， respectively. HRT18 cells were cultured and divided into a blank group and PPI treatment groups with concentrations of 1.2， 1.4， 1.6 μmol·L^-1， respectively. The effects of PPI on the proliferation and morphology of colorectal cancer were detected by cell proliferation toxicity assay， trypan blue exclusion assay， plate clone formation assay， and confocal high-intension cell imaging analysis system. Flow cytometry was used to detect the apoptosis rate of colorectal cancer cells. The pQCXIP-GFP-LC3 plasmid transfection assay was used to detect the formation of autophagosomes in colorectal cancer cells after PPI treatment. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3， Caspase-8， and poly ADP ribose polymerase （PARP）， the expression of autophagy related protein LC3Ⅱ， and the expression and phosphorylation of Hippo signaling pathway proteins LATS1 and YAP. In the plvx-Flag-YAP plasmid transfection assay， YAP was overexpressed and treated with PPI， and the proliferation of colorectal cancer cells was detected by cytotoxicity assay. The expression of LC3Ⅱ and PARP in colorectal cancer cells was detected by Western blot. SwissADME predicted pharmacokinetic parameters of PPI. ResultAs compared with the blank group， the survival rate and clone formation ability of colorectal cancer cells in the PPI group were significantly decreased （P<0.01）， the cell area of colorectal cancer cells in the PPI group was significantly decreased， and the roundness of colorectal cancer cells was significantly increased （P<0.01）. As compared with the blank group， the apoptosis rate of colorectal cancer cells in PPI treatment groupw was significantly increased （P<0.01）， the expression of apoptotic proteins Caspase-3 and Caspase-8 protein precursor in PPI treatment groups was decreased， and the cleavage of PARP was increased （P<0.01）. As compared with the blank group， the expression level of autophagy-related protein LC3Ⅱ in colorectal cancer cells in PPI treatment groups was significantly increased， and the formation of autophagosomes was promoted （P<0.01）. As compared with the blank group， the expression of YAP protein in colorectal cancer cells in PPI treatment groups was significantly decreased， and the expressions of phosphorylated LATS1 and YAP were significantly increased （P<0.01）. As compared with the blank group， overexpression of YAP could significantly antagonize the effect of PPI on apoptosis， autophagy activation， and proliferation inhibition of colorectal cancer cells. SwissADME simulation results showed that PPI had good drug like activity. ConclusionPPI can induce apoptosis and autophagy of colorectal cancer cells through targeted activation of Hippo signaling pathway， thereby inhibiting their proliferation.

Objective:To analyze the cause and epidemiological characteristics of an outbreak of cutaneous anthrax in Caoxian County, Heze City, Shandong Province, and to provide scientific basis for anthrax prevention and control.Methods:Using on-site epidemiological investigation methods and the "Anthrax Epidemiological Case Investigation Form", case investigations were conducted based on the epidemiological contact history and close contacts of suspected anthrax cases reported by the national health care system ( n = 83). Scorched skin smears, diseased cattle tissues, soil samples from the slaughter site and smears from slaughter utensils were collected from cases for Real-time PCR testing and pathogenic bacteria isolation and culture, respectively. Anthrax determination criteria were carried out with reference to "Anthrax Diagnosis" (WS 283-2020). Results:A total of 13 cases of cutaneous anthrax were found in this outbreak, including 12 clinically diagnosed cases and one confirmed case (positive Real-time PCR test and isolation of a strain of Bacillus anthracis). The epidemiological investigation determined that the source of infection in this outbreak was diseased cattle, the transmission route was through slaughter of diseased cattle, contact with contaminated utensils and related cattle products, and the patients were mainly engaged in occupations related to cattle slaughter or cattle product collection and sale. A total of 84 samples were collected, including 13 skin scabs, 64 environmental samples and 7 beef samples. Thirty-six positive PCR tests were performed, with a positive rate of 42.86% (36/84). Among them, 100.00% (13/13) were positive for skin scab smear specimens, 29.69% (19/64) for environmental samples and 4/7 for beef samples. A total of 8 strains of Bacillus anthracis were isolated, including 6 environmental specimens, 1 suspected case and 1 beef strain, with an overall detection rate of 9.52% (8/84). Eighty-three close contacts were investigated. Thirteen households involved in the epidemic were disinfected by spraying (200 ml/m 2) with chlorine-containing disinfectant (5 000 mg/L), and a total of 40 households involved in the epidemic were disinfected, covering an area of about 10 765 m 2. Forty-five pieces of suspected contaminated clothing were burned and disposed of, and 152 pieces of kitchenware were soaked. Conclusions:Slaughter of infected cattle, contact with contaminated utensils and related cattle products are the main causes of this skin anthrax outbreak. Strengthening market supervision, deepening inter-animal epidemic prevention, carrying out publicity and education on anthrax prevention and control, and enhancing practitioners' awareness of disease prevention is the key to prevent anthrax from occurring.

Objective:To evaluate the efficacy and safety of a China-made disposable peroral cholangioscope in animal cholangioscopy.Methods:Six healthy Bama minipigs underwent the domestic disposable peroral cholangioscopy after anesthesia. The operation performance and image quality of the products were evaluated, and intraoperative and postoperative complications were recorded.Results:All 6 animals received the domestic disposable peroral cholangioscopy. The cholangioscope showed good operating performance and could smoothly enter the biliary tract through the duodenoscope. The water injection, suction and instrument channels were unobstructed. The cholangioscopic image was clear, the color resolution was good, and there was no image distortion. The lumen and the mucosal surface could be accurately assessed. During the examination, there was no operative injuries such as bleeding, perforation, or adverse events such as respiratory depression or cardiac arrest. All pigs survived the operation with no adverse reaction.Conclusion:The China-made disposable peroral cholangioscope is safe, with good operability and high-quality image.

Objective:To analyze the global research hotspots in the field of pediatrics based on the Essential Science Indicators (ESI) database and explore the inspiration to domestic editors and pediatrics researchers.Methods:The journal distribution, country (region) distribution, cooperation, organization distribution, funding, publication language, hot topic words and other data of highly cited papers in the field of pediatrics in ESI database were collected and analyzed.Results:A total of 682 highly cited pediatrics papers were collected from 77 pediatrics journals included in Science Citation Index(SCI). Most of the highly cited pediatrics papers (182) were found to be published in Pediatrics.All 682 paper were published in English and frequently, characterized by multiple authors, institutions and fund support.Of 682 highly cited pediatrics papers, 435 papers were published in the United States(the first), 123 papers in England(the second) and 86 paper in Canada(the third). Novel coronavirus pneumonia, coronavirus, SARS coronavirus, autism and multiple system inflammatory syndrome are the main frontiers of global pediatric research at present.Specifically, focal pediatric system diseases mainly include respiratory system diseases, digestive system diseases, cardiovascular diseases, etc. Conclusions:ESI-based analysis of global research hotspots in the field of pediatrics provides reference materials for domestic and foreign pediatrics researchers to understand the global academic frontiers and development trends in the field of pediatrics and select topics for future scientific research.More importantly, this analysis can help domestic editors of pediatrics journals to plan topics and organize hot papers, so as to improve the academic quality and international influence of the journals.

Objective:To study the risk factors for complications after endoscopic retrograde cholangiopancreatography (ERCP) in super-aged patients (≥80 years).Methods:Clinical data of 512 super-aged patients with pancreaticobiliary diseases who underwent 638 ERCP procedures at the Digestive Endoscopy Center, Zhongnan Hospital of Wuhan University, from July 2011 to June 2021, were studied retrospectively. Indications and results of the ERCP operations were analyzed. Multivariate logistic regression model was used to analyze the risk factors for ERCP-related complications.Results:The total success rate of ERCP cannulation in super-aged patients was 94.0% (600/638), which showed no difference compared with that of patients of <60 years old (2 433/2 557, 95.2%) or patients of 60~<80 years old (2 815/3 004, 93.7%) ( χ2=5.49, P=0.064). The overall incidence of post-ERCP complications was 15.2% (97/638), and the in-hospital mortality was 2.1% (11/512), which showed significant difference compared with patients of <60 years old (8/1 809, 0.4%) and patients of 60-<80 years old (21/2 127, 1.0%) ( χ2=13.39, P=0.002). Multivariate regression analysis showed that hypertension ( HR=1.94, 95% CI: 1.237-3.041, P=0.004), history of upper gastrointestinal reconstruction ( HR=2.28, 95% CI: 1.064-4.891, P=0.034), endoscopic sphincterectomy ( HR=1.65, 95%CI: 1.012-2.679, P=0.045), early procedure period ( HR=0.57, 95% CI: 0.352-0.923, P=0.022), operation time >30 minutes ( HR=1.74, 95% CI: 1.094-2.759, P=0.019), preoperative white blood cell count >9.5×10 9/L ( HR=2.66, 95% CI: 1.661-4.257, P<0.001) and procalcitonin ≥0.05 ng/L ( HR=2.54, 95% CI: 1.172-5.513, P=0.018) were independent risk factors for post-ERCP complications. Conclusion:ERCP is safe and effective for super-aged patients. However, much attention should be paid to post-ERCP complications of patients with hypertension, history of upper gastrointestinal reconstruction, endoscopic sphincterectomy, operation time >30 minutes, preoperative white blood cell count >9.5×10 9/L and procalcitonin ≥0.05 ng/L to avoid serious adverse events such as mortality.

Objective:To assess the nursing care experiences and curative effects of the application of wound care technologies combined with other multiple treatments in the hepatolithiasis patient who suffered from postoperative severe intestinal fistula.Methods:The wound care difficult points were analyzed,the wound care technologies combined with other multiple treatments including multiple debridement methods were applied according to the severity of the intestinal fistula:multiple debridement, new skin protection, continuous flushing and drainage with double cannula, improved negative pressure closure drainage and wound pull-in technology, combined with systemic treatment and nursing intervention.Results:The intestinal fistula was treated for 12 days, and the wound infection was controlled for 24 days. Finally the wound was healed after 43 days and patient was discharged from hospital.Conclusions:In the case of severe intestinal fistula after hepatolithiasis operation, the application of wound nursing technology combined with treatment scheme provides a new treatment idea and scheme for intestinal fistula and wound healing.

Objective:To observe the application effect of Rothman-Keller risk forewarning model on stroke incidence in elder community patients with hypertensive.Methods:A total of 1 267 elder hypertensive patients were selected as research objects from 4 community hospitals cooperated with Xinxiang Central Hospital in June 2018. 12 related risk factors were collected according to the Rothman-Keller model, including systolic blood pressure, diastolic blood pressure, high density liptein cholesterol (HDL-c) , low density lipoprotein (LDL-c) , homocysteine (Hcy) , Hyperuricemia, impaired fasting glucose/diabetes, family history of stroke, carotid plaque positive, regular medication, physical exercise and smoking. The differences on risk factors were compared among patients with different characteristics. The patients were followed up in June 2019 and June 2020 respectively. The follow-up mainly included blood pressure control, stroke incidence and death. Rothman-Keller model was used to evaluate the risk of stroke in elderly patients with hypertension in the general population and different gender groups, and the application effect of the model was analyzed.Results:Among 1 267 patients with hypertension, 54 cases were lost to follow-up, and the loss rate was 4.26%. The follow-up time was (43.40±8.22) months. The systolic blood pressure, LDL-c, the proportion of smoking and the positive rate of carotid plaque in male patients were significantly higher than those in female patients, with statistical significance ( P<0.05) . The sensitivity, specificity and diagnostic coincidence rate of Rothman-Keller model for stroke in general population, male population and female population were 0.82, 0.76, 75.93%; 0.86, 0.82, 81.78%; 0.73, 0.83, 82.61%, respectively. According to Rothman-Keller model, the area under the receiver operating characteristic (ROC) curve of stroke in total population, male population and female population were 0.773, 0.784 and 0.824 respectively ( P<0.01) . Conclusions:Rothman-Keller model can well predict the risk of stroke in elderly patients with hypertension in the community. It is a simple and practical prediction tool.

OBJECTIVE: To investigate the phylogenetics and prevalence of bloodstream infections with ST131, the antimicrobial resistance profiles of the pathogens, and the clinical features. METHODS: Non-duplicate isolates were collected from 144 patients with bloodstream infections in our hospital between January and December, 2016.The phylogenetic groups of the isolates were analyzed using multiplex PCR, and O serotyping of ST131 strains was performed by allele-specific PCR.The clinical characteristics of the 144 patients were analyzed to define the differences in the clinical features between patients with ST131 infection and those with non-ST131 infection.Antibiotic susceptibility of the isolates was determined using the Vitek 2 compact system. RESULTS: The phylogenetic group analysis showed a domination by group B2 (41.0%[59/144]), followed by group F, group B1 and group E, which accounted for 16.7%(24/144), 13.9%(20/144), and 13.2% (19/144), respectively.Nine strains (6.3%) of were identified to be ST131 strains, among which 8 were O25b-B2-ST131 strains and 1 was O16-B2-ST131 strain.Of the 9 cases of ST131 infection, 7(77.8%) were found to occur in a nosocomial setting.The demographic characteristics and clinical features of the ST131-infected patients were similar to those of non-ST131-infected patients.ST131 strains were sensitive to piperacillin/tazobactam, imipenem, ertapenem, and amikacin, but showed high resistance rates to cefazolin, ceftriaxone, ciprofloxacin, levofloxacin, gentamicin, and trimethoprim/ sulfamethoxazole (all over 50%).The positivity rate of ESBLs in the ST131 strains was 77.8%, and the multidrug resistance rate reached 88.9%, which was higher than that of non-ST131 isolates, but the difference was not statistically significant. CONCLUSIONS The most common phylogenetic groups of isolates from patients with bloodstream infections are group B2 and F, and the positivity rate of ST131 is low.We for the first time detected O16-ST131 in patients with blood-borne infections in China.The clinical features of ST131-infected patients are similar to those of non-ST131-infected patients.The positivity rate of ESBLs and the multidrug resistance rate are high in ST131 strains, which may raise concerns in the future.
Anti-Bacterial Agents ; therapeutic use ; Bacteremia ; drug therapy ; epidemiology ; microbiology ; China ; Drug Resistance, Bacterial ; Escherichia coli ; classification ; drug effects ; genetics ; Escherichia coli Infections ; drug therapy ; epidemiology ; microbiology ; Genotype ; Humans ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Phylogeny ; Species Specificity