ObjectiveTo study the characteristics of the five tones of Parkinson's disease （PD） patients based on the theory of "five-viscera phonology"， and provide references for the syndrome differentiation and treatment of PD. MethodsA total of 272 cases of PD patients were collected as the PD group， and 240 individuals， including patient family members and hospital staff， were recruited as the control group. The 25-tone analyzer was used to collect the five-tone characteristics of both PD patients and control group participants. The participants were then stratified into three age groups， 41~55， 56~70， and 71~85 years old， and categorized by gender （male and female） for analysis. The frequency and composition ratio of the five tones were analyzed for both groups across the different age ranges and genders. Additionally， the average voice frequency of each participant was calculated to compare differences between groups， stratified by age range and gender. ResultsIn the 41~55 and 56~70 age groups， male participants in the PD group and the control group exhibited the highest frequency of Yu （羽） tone， with the PD group showing a significantly higher composition ratio of Yu tone compared to the control group （P＜0.05）； for males in the 56~70 age group， the composition ratios of Shang （商） and Zhi （徵） tone in the PD group were lower than those in the control group （P＜0.05）. For males in the 71~85 age group， both the PD group and the control group had the highest frequency of Yu tone， but there was no statistically significant difference in the composition ratios of the five tones between groups （P＞0.05）. For female participants in the PD group across all age groups， Yu tone was the most frequent， whereas for the control group， Jue （角） tone was the most frequent in all age groups， and the composition ratio of Yu tone in the PD group was significantly higher than that in the control group across all age groups （P＜0.05）； in the 56~70 age group， the composition ratio of Jue tone was lower in the PD group compared to that in the control group （P＜0.05）. Regarding voice frequency， males in the PD group aged 41~55 and 56~70 had higher voice frequency than those in the control group of the same age range， and similarly， females in the PD group aged 56~70 and 71~85 had higher voice frequency than their counterparts in the control group （P＜0.05）. ConclusionPD patients have a voice with a higher frequency and an increased proportion of Yu tone in their five-tone distribution. According to the theory of five-viscera phonology， PD patients may have disease mechanism of kidney essence deficiency.

BACKGROUND:Primary cortical neurons and microglial cells play a crucial role in exploring cell therapies for neurological disorders,and most of the current methods for obtaining the two types of cells are cumbersome and require separate extraction.It is therefore crucial to find a convenient and rapid method to extract both types of cells simultaneously. OBJECTIVE:To explore a novel method for simultaneous extraction of primary cortical neurons and microglial cells. METHODS:Newborn suckling SD rats were taken within 24 hours.The brain was removed and placed in a dish with DMEM,and the pia mater was removed for later use.Primary neurons were extracted from the same brain tissue,and then the remaining brain tissue was used to extract microglial cells.The whole process was performed on ice.Extraction and culture steps of primary cortical neurons:The cerebral cortex was taken 2.0-3.0 mm with forceps,and the tissue was digested with papain for 20 minutes.After aborting digestion,the blown tissue presented an adherent tissue suspension.The supernatant cell suspension was obtained,filtered,and dispensed into 15 mL centrifuge tubes.After centrifugation and re-suspension,the cells were inoculated onto 6-well plate crawls coated with L-polylysine.Neuronal morphology was observed at 1-day intervals,and staining could be performed for identification using immunofluorescence staining of MAP2 and β-Tubulin by day 7.Microglia extraction and culture steps:The remaining brain tissue at 8-10 mm thick was subjected to microglial cell extraction,digested by trypsin for 20 minutes.After digestion was stopped,the tissue was blown to a homogenate,and then the homogenate was transferred to the culture bottle for culture.On day 14,the culture flasks were sealed and subjected to constant temperature horizontal shaking for 2 hours.Microglial cells were shed in the supernatant.Purified microglial cells were taken and continued to be cultured for 3 days for identification by Iba1 immunofluorescence staining. RESULTS AND CONCLUSION:(1)After 24 hours of culture,the neurons were adherent to the wall,the cytosol was enlarged,and some neurons developed synapses.After 3 and 5 days of culture,the cytosol was further enlarged,and most of the neurons were in the form of synapses,and some neurons were growing in clusters.On day 7,neuronal synapses were prolonged and thickened,and they were connected with each other to form a network.The neurons were identified by β-Tubulin and MAP2 immunofluorescence staining.(2)The cells grew close to the wall on day 1 of culture.On days 3,5,and 7,the density of microglial cells was small,and the cell morphology was bright oval or round,but the cells basically grew in clumps on the upper layer of other cells.On day 10,the density of microglial cells increased significantly.On day 14,microglial cells grew in dense clumps on the upper layer of other cells,and then they could be isolated and purified.The isolated and purified cells were taken and re-cultured to day 3 and identified as microglial cells by Iba1 immunofluorescence;their purity was greater than 95%.(3)The results show that primary cortical neurons and microglial cells obtained by this method after extraction and culture are of high purity,good morphology,and high viability.

Objective To analyze signals of adverse events(AEs)related to macrolide antibiotic in children from the US FDA Adverse Event Reporting System(FAERS),and provide a reference for clinical safe medication in children.Methods The AEs reports of children(less than 18 years old)were extracted from FAERS during January 2004 to March 2024,and the reports of odds ratio(ROR)and information component of Bayesian confidence propagation neural network(IC)were used to analyze the signals of macrolide antibiotics in children.Results There were 2 133 648 reports of children from FAERS,including azithromycin(7 589 cases),clarithromycin(3 673 cases),and erythromycin(820 cases).The significant signals of system organ class(SOC)were 16 in total,such as skin disorders,ear and labyrinth disorders,gastrointestinal disorders,nervous system,immune system disorders,etc.A proportional imbalance analysis was performed on macrolide antibiotics(azithromycin and clarithromycin),and obtained relevant positive signals,including azithromycin(232)and clarithromycin(194).The most frequently reported AEs included rash,vomiting,diarrhea,hypersensitivity and so on.The strongest signals of azithromycin were Stevens-Johnson syndrome(SJS),hypersensitivity reactions,drug-induced liver injury,etc.,while the strongest signals of clarithromycin were dysgeusia,lip swelling,toxic epidermal necrolysis(TEN),nightmare,psychotic disorder,etc.17 unlisted signals of azithromycin and 21 of clarithromycin were found.The same PT of azithromycin and clarithromycin had significant difference,such as dyspnea,hypersensitivity,rash,SJS,vomiting,and more.Conclusions To ensure the safety of children's treatment,children using macrolide should pay attention to skin,gastrointestinal,ear and other toxicity and hypersensitivity reactions,especially serious skin reactions,unlisted suspicious AEs and psychiatric disorders.

The elevated polyamines, amine-rich molecules with diverse functions in pathophysiology processes, are implicated in contributing to tumorigenesis and progression. Whether and how they affect the efficacy of chemotherapy is incompletely understood. Our screening assays reveal that the supplement with a low dose of spermidine (Spd), one of the polyamines, enhances ferroptosis in prostate cancer cells as evidenced by increased lipid peroxidation and intracellular Fe²⁺ levels in vitro. Combination treatment with Spd and a low dose of ferroptosis inducer erastin synergistically augments anti-tumor efficacy with undetectable toxicity in mice. Analysis of RNA-seq data indicates that heme oxygenase 1 (HMOX1), an enzyme that catalyzes the cleavage of heme to release Fe²⁺, is significantly upregulated in response to Spd and erastin cotreatment. Spd mediated the hypusine modification of the eukaryotic initiation factor 5A (EIF5A) promotes the translation of the nuclear factor erythroid 2-related factor 2 (NRF2), subsequently leading to elevation of HMOX1. Moreover, Spd and erastin significantly inhibit proteasome activity which results in a decrease in proteasomal degradation of NRF2, although many proteasome-related genes are induced either by Spd or Spd plus erastin. Thus, in addition to its pro-oncogenic activity, the supplement of Spd improves antitumor activity in combination with ferroptosis inducers and offers an optional approach to cancer treatment.

In recent years, studies have found that the abnormal expression of rhythm genes is closely related to the risk of occurrence and the progression of the disease course of colorectal cancer. In addition, chronotherapy based on the circadian rhythm theory has shown certain effects in the clinical treatment of colorectal cancer, but there are still great limitations. Therefore, it is very important to clarify the mechanism of action of rhythm genes in the occurrence and development of colorectal cancer, which may provide a theoretical basis for the clinical application of chronotherapy.

Objective To explore the influencing factors of depression risk in elderly patients with hypertension and construct and validate a nomogram prediction model.Methods A total of 869 elderly patients with hypertension were selected from national survey database of the China Health and Retirement Longitudinal Study(CHARLS)in 2018.Multivariate Logistic regression analysis was used to identify the risk factors for depression in elderly patients with hypertension,and a nomogram prediction model was constructed.The accuracy and effectiveness of the model were validated by the Hosmer-Lemeshow(H-L)goodness-of-fit test,the area under the curve(AUC)of the receiver oper-ating characteristic(ROC)curve,and the calibration curve.Results The incidence of depression in elderly patients with hypertension was 47.18％.Factors influencing the risk of depression included rural residence(OR=2.191,P＜0.05),impaired basic activities of daily living(BADL)(OR=2.338,P＜0.05),impaired instrumental activitiesofdaily living(IADL)(OR=1.674,P＜0.05),poor life satisfaction(OR=7.348,P＜0.05),fair self-rated health(OR=0.441,P＜0.05),good self-rated health(OR=0.259,P＜0.05),and sleep duration of 6 to 9 hours(OR=0.510,P＜0.05).The AUC of the ROC curve was 0.795,the slope of the calibration curve was close to 1,and the H-L goodness-of-fit test yielded x2=5.074.The validation set showed an AUC of 0.703.Conclusion The prediction model established in this study has high accuracy and discriminative ability.Healthcare professionals can take effective preventive measures based on individual patient factors.

Objective:The aim of this study is to analyze the clinical characteristics and genetic variants of a late-onset auditory neuropathy pedigree caused by maternally inherited- mitochondrial mutation.Methods:A male proband who presented with bilateral sensorineural hearing loss at Henan Children′s Hospital in September 2023 was chosen, along with his family members (4 generations, 20 individuals) as the study subjects. Data from this pedigree were collected, organized, and analyzed for clinical genetic characteristics. Medical histories were obtained from family members, pedigree charts were drawn, audiological, imaging, and physical examinations were conducted. Pathogenic genes and mutations were screened using high-throughput sequencing. Sanger sequencing was employed for variant confirmation and segregation validation in the family.Results:In this family, a total of 12 members (10 members collected) had sensorineural hearing loss, characterized by late-onset hearing impairment with an onset age ranging from 9 to 30 years. The patients exhibited poor speech recognition rates, and audiometric examinations are consistent with auditory neuropathy. There was no history of ototoxic drug use. High-throughput sequencing identified the variant NC_012920.1:m.7471dup in the mitochondrial MT-TS1 gene as the pathogenic variant. Sanger sequencing results confirmed that the pathogenic gene mutation site perfectly co-segregated with the auditory neuropathy phenotype in this family. According to the classification criteria and guidelines for genetic variations by the American College of Medical Genetics and Genomics, the variant was classified as a pathogenic mutation. Conclusion:The mitochondrial MT-TS1 gene mutation m.7471dup is considered to be the pathogenic cause in this late-onset auditory neuropathy pedigree.

Objective To analyze signals of adverse events(AEs)related to macrolide antibiotic in children from the US FDA Adverse Event Reporting System(FAERS),and provide a reference for clinical safe medication in children.Methods The AEs reports of children(less than 18 years old)were extracted from FAERS during January 2004 to March 2024,and the reports of odds ratio(ROR)and information component of Bayesian confidence propagation neural network(IC)were used to analyze the signals of macrolide antibiotics in children.Results There were 2 133 648 reports of children from FAERS,including azithromycin(7 589 cases),clarithromycin(3 673 cases),and erythromycin(820 cases).The significant signals of system organ class(SOC)were 16 in total,such as skin disorders,ear and labyrinth disorders,gastrointestinal disorders,nervous system,immune system disorders,etc.A proportional imbalance analysis was performed on macrolide antibiotics(azithromycin and clarithromycin),and obtained relevant positive signals,including azithromycin(232)and clarithromycin(194).The most frequently reported AEs included rash,vomiting,diarrhea,hypersensitivity and so on.The strongest signals of azithromycin were Stevens-Johnson syndrome(SJS),hypersensitivity reactions,drug-induced liver injury,etc.,while the strongest signals of clarithromycin were dysgeusia,lip swelling,toxic epidermal necrolysis(TEN),nightmare,psychotic disorder,etc.17 unlisted signals of azithromycin and 21 of clarithromycin were found.The same PT of azithromycin and clarithromycin had significant difference,such as dyspnea,hypersensitivity,rash,SJS,vomiting,and more.Conclusions To ensure the safety of children's treatment,children using macrolide should pay attention to skin,gastrointestinal,ear and other toxicity and hypersensitivity reactions,especially serious skin reactions,unlisted suspicious AEs and psychiatric disorders.

Objective:The aim of this study is to analyze the clinical characteristics and genetic variants of a late-onset auditory neuropathy pedigree caused by maternally inherited- mitochondrial mutation.Methods:A male proband who presented with bilateral sensorineural hearing loss at Henan Children′s Hospital in September 2023 was chosen, along with his family members (4 generations, 20 individuals) as the study subjects. Data from this pedigree were collected, organized, and analyzed for clinical genetic characteristics. Medical histories were obtained from family members, pedigree charts were drawn, audiological, imaging, and physical examinations were conducted. Pathogenic genes and mutations were screened using high-throughput sequencing. Sanger sequencing was employed for variant confirmation and segregation validation in the family.Results:In this family, a total of 12 members (10 members collected) had sensorineural hearing loss, characterized by late-onset hearing impairment with an onset age ranging from 9 to 30 years. The patients exhibited poor speech recognition rates, and audiometric examinations are consistent with auditory neuropathy. There was no history of ototoxic drug use. High-throughput sequencing identified the variant NC_012920.1:m.7471dup in the mitochondrial MT-TS1 gene as the pathogenic variant. Sanger sequencing results confirmed that the pathogenic gene mutation site perfectly co-segregated with the auditory neuropathy phenotype in this family. According to the classification criteria and guidelines for genetic variations by the American College of Medical Genetics and Genomics, the variant was classified as a pathogenic mutation. Conclusion:The mitochondrial MT-TS1 gene mutation m.7471dup is considered to be the pathogenic cause in this late-onset auditory neuropathy pedigree.

Objective:To investigate the acute gastrointestinal injury (AGI) in patients with extracorporeal membrane oxygenation (ECMO) at the early stage of operation and its influencing factors.Methods:A total of 70 patients with ECMO who were hospitalized in the Emergency Care Unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed, and a total of 70 patients with ECMO who were hospitalized in the emergency care unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed. According to the 2012 guidelines of the European Society of Intensive Care Medicine on the classification of acute gastrointestinal injury in critically ill patients, the patients were divided into AGI group and non-AGI group. The incidence of acute gastrointestinal injury in the early stage was statistically analyzed, and the results of blood gas analysis during ECMO loading and ECMO parameters, hemodynamic indexes and biochemical indexes after ECMO transfer were statistically analyzed. To explore the influencing factors and independent risk factors of AGI in the early stage. In addition, 70 patients were divided into successful group and non-successful group according to whether they were successfully withdrawn. The occurrence of acute gastrointestinal injury between the two groups was compared, and the effect of acute gastrointestinal injury on ECMO patients was analyzed.Results:Among the 70 ECMO patients, the incidence of early AGI was 71.43% (50 cases), and the components of AGI Ⅰ, Ⅱ, Ⅲ and Ⅳ were 18.57% (13 cases), 41.43% (29 cases), 11.43% (8 cases) and 0% (0 cases), respectively. ① Univariate analysis showed that systolic blood pressure, diastolic blood pressure, mean arterial pressure (MAP), vasoactive drug index (VIS), pH, lactic acid and BMI were significantly different between AGI group and non-AGI group when ECMO was used ( P < 0.05). Logistic binary regression analysis showed that BMI was an independent risk factor for early AGI in ECMO patients (ROC area 0.657, 95% confidence interval 0.522-0.791 ( P < 0.05), and Yoden index 0.15). (3) The AGI composition ratio of the unsuccessful group was higher than that of the unsuccessful group ( P < 0.05). Conclusions:Patients with ECMO have a high incidence of AGI in the early stage, mainly occurring in grade I and Ⅱ. Systolic blood pressure, diastolic blood pressure, MAP, VIS, pH, lactic acid and BMI when ECMO is put on are influential factors for the early development of AGI in ECMO patients, among which BMI is an independent risk factor for the early development of AGI in ECMO patients. The occurrence of AGI reduces the probability of successful withdrawal in ECMO patients.