BACKGROUND:In recent years,numerous studies have shown that autophagy and mitophagy play an important role in the regulation of bone metabolism.Under non-physiological conditions,mitophagy breaks the balance of bone metabolism and triggers metabolism disorders,which affect osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells,etc. OBJECTIVE:To summarize the mechanism of mitophagy in regulating bone metabolic diseases and its application in clinical treatment. METHODS:PubMed,Web of Science,CNKI,WanFang and VIP databases were searched by computer using the keywords of"mitophagy,bone metabolism,osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells"in English and Chinese.The search time was from 2008 to 2023.According to the inclusion criteria,90 articles were finally included for review and analysis. RESULTS AND CONCLUSION:Mitophagy promotes the generation of osteoblasts through SIRT1,PINK1/Parkin,FOXO3 and PI3K signaling pathways,while inhibiting osteoclast function through PINK1/Parkin and SIRT1 signaling pathways.Mitophagy leads to bone loss by increasing calcium phosphate particles and tissue protein kinase K in bone tissue.Mitophagy improves the function of chondrocytes through PINK1/Parkin,PI3K/AKT/mTOR and AMPK signaling pathways.Modulation of mitophagy shows great potential in the treatment of bone diseases,but there are still some issues to be further explored,such as different stages of drug-activated mitophagy,and the regulatory mechanisms of different signaling pathways.

Objective To study the protective effect of Wedelolactone(WEL)against inflammatory injury in human umbilical vein endothelial cells(HUVECs)and its molecular mechanism by inducing PI3K/Akt/mTOR.Methods The model of atherosclerosis(AS)oxidative stress injury in HUVECs was induced with 200 μmol·L-1 of hydrogen peroxide for 24 h.The experimental groups were as follows:normal control group,DMSO(dimethyl sulfoxide)group,H2O2 group,and WEL group.MTT was used to measure the cell survival rate of each group;flow cytometry was used to assess intracellular ROS levels;fluorescence microscopy was used to detect the expression of p62 protein;immunoblotting assay was used to determine the protein expression levels for apoptosis-related proteins associated with PI3K/Akt/mTOR signaling pathway and autophagy-related proteins.Results Compared with the H2 O2 group,the HUVEC cell survival rate was significantly inhibited in the WEL group(P＜0.05).ROS production was significantly lower,and the protein expressions of SOD1 and p62 were significantly increased in the WEL group as compared to the hydrogen peroxide group.The protein expression of p-mTOR,p-Akt,and p-PI3K was significantly decreased in hydrogen peroxide(P＜0.01);In the WEL experiment,p-mTOR,p-Akt,and p-PI3K were increased significantly in the post-injury HUVECs(P＜0.01).Conclusion Wedelolactone inhibits HUVECs'autophagy by suppressing H2O2-induced inflammatory damage in HUVECs,which may be related to the fact that WEL promotes the phosphorylation of PI3K,Akt,and mTOR proteins,inhibits autophagy and thus resists oxidative stress damage in HUVECs cells.

Objective:To observe the 95% effective dose (ED95) of Propofol mono-sedation for successfully inserting the gastroscope in healthy adults by biased coin design up-and-down sequential method.Methods:Using prospective study method, a total of 40 patients proposed for painless gastroscopy in Beijing Friendship Hospital, Capital Medical University from April to May 2021 were selected. There were 15 males and 25 females. American Society of Anesthesiologists (ASA) classification: grade I 26 cases, grade Ⅱ 14 cases. The mean age was (50.80±9.14) years, and the mean body mass index was (24.08±2.65) kg/m 2. Propofol mono-sedation was used in all patients. The initial dose of Propofol was set as 1.6 mg/kg, adjusted with 0.1 mg/kg as a step size. The biased coin design up-and-down sequential method was used in this study. The Propofol dose of subsequent patients was determined by the response to gastroscope insertion of the previous patient. If the gastroscopy insertion reaction of the previous patient was positive, the Propofol dose of the next patient was increased by one level (0.1 mg/kg); if the gastroscopy insertion reaction of the previous patient was negative, the biased coin random was performed, and the Propofol dose used by the next patient was reduced by one level (0.1 mg/kg) with 5% probability and remained unchanged with 95% probability. Changes of mean arterial pressure, heart rate and pulse oxygen saturation were recorded at different time points, and adverse reactions such as perioperative hypotension, bradycardia, tachycardia and hypoxemia were recorded. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test was used for comparison between different time points. The ED95 and 95% CI of Propofol in inhibiting the response to gastroscope insertion was calculated by Probit regression analysis. Results:All 40 patients successfully completed the gastroscopy. The calculated ED95 of Propofol mono-sedation for successfully inserting the gastroscope was 2.58 mg/kg with 95% CI of 2.40-3.31 mg/kg. The mean arterial pressure before anesthesia, after propofol injection, at the time of gastroscopy going through throat and immediately after examination was (97.33±13.34) mmHg, (93.15±11.35) mmHg, (78.95±9.30) mmHg, (79.38±9.94) mmHg (1 mmHg=0.133 kPa), respectively. The mean arterial pressure at the time of gastroscopy going through throat and immediately after examination decreased significantly, the difference was statistically significant ( P<0.01). There were no significant differences in heart rate and pulse oxygen saturation compared with those before anesthesia ( P>0.05). Conclusion:The ED95 of Propofol mono-sedation for successfully inserting the gastroscope is determined as 2.58 mg/kg (95% CI: 2.40-3.31 mg/kg) by biased coin design up-and-down sequential method.

Objective To investigate the effect of Yu's Qingre Huoxue prescription enema on colonic mucosa in rats with ulcerative colitis(UC).Methods A total of 30 Wistar rats were randomly divided into blank control group,model group,mesalazine group,Chinese medicine high-dose,medium-dose and low-dose groups(5 rats in each group).After successful modeling with trinitrobenzene sulfonic acid,the rats were given 14 days enema with distilled water,mesalazine suspension,and high,medium and low concentration Yu's Qingre Huoxue prescription.The general condition and weight changes of rats were recorded daily,and colon macroscopic damage index(CMDI)and mucosal injury score under microscope of rats in each group were compared.Results Compared with blank control group,the weight of rats in model group decreased significantly(P<0.05).The blank control group had normal water and food intake and activity,glossy fur and normal stool.The rats in model group and each administration group were found to have anal fouling,residual blood in stool,decreased appetite,messy fur,dim luster and other conditions,and the above conditions were gradually improved after enema treatment.Compared with blank control group,the CMDI and mucosal injury scores of model group were significantly increased(P<0.05).Compared with model group,CMDI and mucosal injury scores of mesalazine group and Chinese medicine high-dose,medium-dose and low-dose groups were significantly decreased(P<0.05).Conclusion Yu's Qingre Huoxue prescription can effectively improve the clinical symptoms of diarrhea,mucous,pus and blood stools in UC model rats,reduce the damage of colonic mucosa,promote mucosal repair,and have a good effect on the treatment of UC in rats.

Objective To rationally allocate pharmaceutical and healthcare human resources,and to explore the feasibility of implementing the system of licensed assistant pharmacists in retail pharmacies.Methods Through literature review,questionnaire surveys,and statistical analysis of exam scores,the training status of pharmacy technicians with secondary vocational education,the employment situation in retail pharmacies,and the results of the licensed pharmacist qualification examination were deeply understood.Results Every year,a certain number of graduates with a secondary vocational education in pharmacy entered society,and retail pharmacies were willing to recruit pharmacy technicians with this educational background,their professional competence were recognized.Pharmacy technicians with secondary vocational education can obtain the professional qualification of practicing pharmacists through hard work,and demonstrated the basic competency for working as licensed assistant pharmacists.Conclusion The implementation of the licensed assistant pharmacist system among pharmacy technicians with secondary vocational education in retail pharmacies is feasible.This system can fully leverage the role of pharmacy technicians with secondary vocational education,make up for the shortage of professional personnel in certain regions,promote the construction of the pharmaceutical care system in retail pharmacies,and ensure drug quality and medication safety.

Objective To investigate the design of cuffed pediatric tracheal tubes and compare the effects of different tracheal intubation depth placement strategies on the position of the tracheal tube tip and cuff of 5 tracheal tube brands.Methods A total of 180 children who were admitted to Tianjin Children's Hospital from October 2020 to December 2021,with endotracheal intubation under general anesthesia,aged 1-6 years,were enrolled.The length of the subglottic airway was measured by electronic bronchoscopy.Dimensional data from 5 cuffed pediatric tracheal tube brands were recorded,including the length of the tracheal tube cuff,the distance from the tip of the tracheal tube to the upper edge of the cuff,and the tip of the tracheal tube to the lower edge of the tube glottis marker line the distance.Calculation of the required cuffed endotracheal tube internal diameter(ID)for 180 pediatric patients was performed based on the Motoyama formula,the positions of tracheal tube tip and upper cuff border were calculated for each of the 180 tracheas using depth mark to based tracheal tube placement,placement of the tracheal tube tip at 2 cm above the carina,and mid-tracheal tube placement.Results There were differences in the dimensional data of the 5 cuffed pediatric tracheal tube brands.Depth mark-based tracheal tube placement resulted in the incidence rate of tube tip to carina placement less than 1 cm was 3.9%-67.8%,and the highest incidence of bronchial intubation is Ruijing,up to 17.8%.The tracheal tube tip was placed 2 cm above the carina,and no improper placement of the tracheal tube cuff and tube tip was found in all brands.Mid-tracheal tube placement led to 100%subglottic and supraglottic tracheal tube cuff positions,except Weili.Conclusions There are differences in design between different brands of cuffed pediatric tracheal tube,and some of the design deficiencies may lead to the risk of airway complications.The method used to guide the insertion depth and the brand of cuffed tracheal tubes can affect the tracheal tube position.Placement of the tracheal tube tip at 2 cm above the carina allowed safe tracheal tube placement in children aged 1-6 years.

Objective:This study aimed to explore the clinical value of myostatin(MST) and follistatin(FST) as biological biomarkers in evaluating sarcopenia in elderly women.Methods:This was a retrospective cross-sectional study that enrolled 350 females aged 20-89 years who underwent physical examinations in Shanghai Huadong Hospital in 2021. Demographic characteristics, muscle mass, fat mass, bone mineral density, hand grip strength, gait speed, and serum indices of MST and FST were collected.Results:The serum levels of MST did not change significantly with age. However, the serum levels of FST increased with age. In women aged≥60 years, MST was positively correlated with total lean mass and appendicular skeletal muscle index(ASMI; r=0.236, P=0.041; r=0.289, P=0.014), while FST was negatively correlated with ASMI( r=-0.265, P=0.030). In multivariate stepwise regression analysis, after adjusting for age, body mass index, hip bone mineral density, and total fat mass, only FST was independently correlated with ASMI( β=-0.238, P=0.006), while MST was not correlated with ASMI. The receiver operating characteristic curve was plotted using muscle mass reduction as the state variable and serum FST level as the test variable. The area under the curve was 0.753. And when the FST cutoff value was 17.49 ng/mL, the maximum Jordan index was 0.46, with a sensitivity of 77.3% and a specificity of 68.7%. Women aged ≥60 years were divided into three groups based on serum FST levels. Compared to the upper third of the serum FST level group, the low third of the FST level group had a significantly reduced risk of suffering from sarcopenia( OR=0.098, P =0.036). Conclusions:Serum FST lever has a better correlation with muscle mass among elderly women, making it a promising biomarker for evaluating muscle mass.

OBJECTIVE: To investigate the effect of inhibitor of growth protein-2 (Ing2) silencing on angiotensin Ⅱ (AngⅡ)-induced cardiac remodeling in mice and explore the underlying mechanism. METHODS: An adenoviral vector carrying Ing2 shRNA or empty adenoviral vector was injected into the tail vein of mice, followed 48 h later by infusion of 1000 ng · kg^-1 · min^-1 Ang Ⅱ or saline using a mini-osmotic pump for 42 consecutive days. Transthoracic echocardiography was used to assess cardiac geometry and function and the level of cardiac hypertrophy in the mice. Masson and WGA staining were used to detect myocardial fibrosis and cross-sectional area of cardiomyocytes, and myocardial cell apoptosis was detected with TUNEL assay. Western blotting was performed to detect myocardial expressions of cleaved caspase 3, ING2, collagen Ⅰ, Ac-p53(Lys382) and p-p53 (Ser15); Ing2 mRNA expression was detected using real-time PCR. Mitochondrial biogenesis, as measured by mitochondrial ROS content, ATP content, citrate synthase activity and calcium storage, was determined using commercial assay kits. RESULTS: The expression levels of Ing2 mRNA and protein were significantly higher in the mice with chronic Ang Ⅱ infusion than in saline-infused mice. Chronic infusion of AngⅡ significantly increased the left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD) and reduced left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) in the mice. Ing2 silencing obviously alleviated AngⅡ-induced cardiac function decline, as shown by decreased LVEDD and LVESD and increased LVEF and LVFS, improved myocardial mitochondrial damage and myocardial hypertrophy and fibrosis, and inhibited cardiomyocyte apoptosis. Chronic AngⅡ infusion significantly increased myocardial expression levels of Ac-p53(Lys382) and p-p53(Ser15) in the mice, and Ing2 silencing prior to AngⅡ infusion lessened AngⅡ- induced increase of Ac-p53(Lys382) without affecting p53 (ser15) expression. CONCLUSION Ing2 silencing can inhibit AngⅡ-induced cardiac remodeling and dysfunction in mice by reducing p53 acetylation.
Animals ; Mice ; Angiotensin II ; Tumor Suppressor Protein p53 ; Acetylation ; Stroke Volume ; Ventricular Remodeling ; Ventricular Function, Left ; Myocytes, Cardiac

The CRISPR-Cas9 system is composed of a clustered regularly interspaced short palindromic repeat (CRISPR) and its associated proteins, which are widely present in bacteria and archaea, serving as a specific immune protection against viral and phage secondary infections. CRISPR-Cas9 technology is the third generation of targeted genome editing technologies following zinc finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs). The CRISPR-Cas9 technology is now widely used in various fields. Firstly, this article introduces the generation, working mechanism and advantages of CRISPR-Cas9 technology; secondly, it reviews the applications of CRISPR-Cas9 technology in gene knockout, gene knock-in, gene regulation and genome in breeding and domestication of important food crops such as rice, wheat, maize, soybean and potato. Finally, the article summarizes the current problems and challenges encountered by CRISPR-Cas9 technology and prospects future development and application of CRISPR-Cas9 technology.
Gene Editing ; CRISPR-Cas Systems/genetics* ; Plant Breeding ; Crops, Agricultural/genetics* ; Technology

OBJECTIVE: To assess the influence of FLT3 expression on the prognosis of patients with acute myeloid leukemia (AML) by cell experiment and clinical data analysis. METHODS: Models for FLT3 over-expression and interference-expression in AML cells were constructed. The level of BAK gene expression and its protein product was determined, along with the proliferation and apoptosis of leukemia cells. FLT3 gene expression and FLT3-ITD variant were determined among patients with newly diagnosed AML. RESULTS: Compared with the interference-expression group, the level of BAK gene expression and its protein in FLT3 over-expression AML cells was significantly lower (P < 0.001), which also showed significantly faster proliferation (P < 0.001) and lower rate of apoptosis (P < 0.001). The expression level of FLT3 gene among patients with newly diagnosed AML was also significantly higher compared with the healthy controls (P < 0.001). The FLT3 gene expression of FLT3-ITD positive AML patients was higher than that of FLT3-WT patients (P = 0.002). Survival analysis showed that AML patients with high FLT3 expression in the medium-risk group had a lower complete remission rate and overall survival rate compared with those with a low FLT3 expression (P < 0.001). CONCLUSION Over-expression of FLT3 may influence the course of AML by promoting the proliferation of leukemia cells and inhibiting their apoptosis, which in turn may affect the prognosis of patients and serve as a negative prognostic factor for AML.
Humans ; Apoptosis/genetics* ; Data Analysis ; Leukemia, Myeloid, Acute/genetics* ; Gene Expression ; fms-Like Tyrosine Kinase 3/genetics*