BACKGROUND:As a new treatment method,local injection of medical ozone has anti-inflammatory,analgesic and immunomodulatory effects on injured skeletal muscle tissue,but its mechanism of action is still lacking systematic research.OBJECTIVE:To observe the intervention effect of medical ozone on skeletal muscle crush injury of rabbit models,and to analyze the differential gene expression of injured rabbit tibialis anterior muscle after ozone treatment through RNA-sequencing and tandem mass tag labeled quantitative proteomic technology,in order to partially reveal the molecular mechanism of medical ozone treatment for skeletal muscle injury.METHODS:Eighteen Japanese white rabbits were randomly divided into a blank group,a model group,and an ozone group,with 6 rabbits in each group.Rabbit tibialis anterior muscle compression injury models were established in the latter two groups.After 12 hours of model establishment,2 mL of medical ozone with a mass concentration of 30 μg/mL was injected into the injured muscle of rabbits in the ozone group.Tissue samples were taken 3 days after ozone treatment.Hematoxylin-eosin staining was used to observe the morphological changes of skeletal muscle.RNA sequencing and tandem mass spectrometry tag were used to detect rabbit tibialis anterior muscle tissue.Bioinformatics analysis was used to explore the biological processes of differentially expressed genes and proteins between groups and then to investigate the molecular mechanism of ozone treatment of skeletal muscle injury.RESULTS AND CONCLUSION:After 3 days of ozone treatment,compared with the blank group,the model group showed swelling of cells,infiltration of inflammatory cells,and partial dissolution of muscle fibers.Compared with the model group,cell edema was alleviated and the number of inflammatory cells was reduced in the ozone group.(2)The results of RNA sequencing and bioinformatics analysis showed that compared with the blank group,596 differentially expressed genes were screened in the model group.Compared with the model group,405 differentially expressed genes were screened in the ozone group.There were a total of 194 differentially expressed genes shared among these differentially expressed genes.Proteomic analysis results exhibited that compared with the blank group,the model group contained 138 differentially expressed proteins.Compared with the model group,242 differentially expressed proteins were determined in the ozone group.There were 66 differentially expressed proteins shared between the two comparison groups.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on differentially expressed genes/proteins,and it was found that they were mainly enriched in the PI3K-Akt signaling pathway,Ras signaling pathway,and MAPK signaling pathway.It is indicated that medical ozone may promote the repair of injured muscle through regulating PI3K/Akt/NF-κB and Ras/MAPK/NF-κB signaling pathway by acting on target genes,such as Syk,FGF16,CSF-1,and MRAS.

Medical therapy and surgical intervention are the two primary approaches for treating myocardial bridge.However,there remains controversy regarding the use of coronary artery bypass grafting(CABG)and myocardial bridge unroofing.Here,we report a case of myocardial infarction following CABG in a patient with a myocardial bridge.The patient was admitted to Lanzhou First Peopie's Hospital with persistent chest pain,chest tightness,and shortness of breath lasting 2 hours.Physical examination revealed no significant abnormalities.Electrocardiography(ECG)indicated extensive anterior wall myocardial infarction.Laboratory findings showed myoglobin levels of 140.1 ng/ml and troponin Ⅰ levels of 2.59 ng/ml,with no other significant abnormalities.The initial diagnosis was acute extensive anterior wall myocardial infarction.Emergency coronary angiography revealed a myocardial bridge in the mid-segment of the left anterior descending artery(LAD).Emergency CABG using the left internal mammary artery to the LAD was performed,leading to symptomatic improvement,and the patient was discharged in stable condition.However,the patient experienced a recurrent myocardial infarction seven years post-surgery and received secondary preventive medical therapy.The patient is currently under ongoing follow-up care.CABG is an effective treatment for myocardial bridge.However,based on the case reported in this study,we recommend careful evaluation of whether a patient may benefit from CABG.

BACKGROUND:As a new treatment method,local injection of medical ozone has anti-inflammatory,analgesic and immunomodulatory effects on injured skeletal muscle tissue,but its mechanism of action is still lacking systematic research.OBJECTIVE:To observe the intervention effect of medical ozone on skeletal muscle crush injury of rabbit models,and to analyze the differential gene expression of injured rabbit tibialis anterior muscle after ozone treatment through RNA-sequencing and tandem mass tag labeled quantitative proteomic technology,in order to partially reveal the molecular mechanism of medical ozone treatment for skeletal muscle injury.METHODS:Eighteen Japanese white rabbits were randomly divided into a blank group,a model group,and an ozone group,with 6 rabbits in each group.Rabbit tibialis anterior muscle compression injury models were established in the latter two groups.After 12 hours of model establishment,2 mL of medical ozone with a mass concentration of 30 μg/mL was injected into the injured muscle of rabbits in the ozone group.Tissue samples were taken 3 days after ozone treatment.Hematoxylin-eosin staining was used to observe the morphological changes of skeletal muscle.RNA sequencing and tandem mass spectrometry tag were used to detect rabbit tibialis anterior muscle tissue.Bioinformatics analysis was used to explore the biological processes of differentially expressed genes and proteins between groups and then to investigate the molecular mechanism of ozone treatment of skeletal muscle injury.RESULTS AND CONCLUSION:After 3 days of ozone treatment,compared with the blank group,the model group showed swelling of cells,infiltration of inflammatory cells,and partial dissolution of muscle fibers.Compared with the model group,cell edema was alleviated and the number of inflammatory cells was reduced in the ozone group.(2)The results of RNA sequencing and bioinformatics analysis showed that compared with the blank group,596 differentially expressed genes were screened in the model group.Compared with the model group,405 differentially expressed genes were screened in the ozone group.There were a total of 194 differentially expressed genes shared among these differentially expressed genes.Proteomic analysis results exhibited that compared with the blank group,the model group contained 138 differentially expressed proteins.Compared with the model group,242 differentially expressed proteins were determined in the ozone group.There were 66 differentially expressed proteins shared between the two comparison groups.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on differentially expressed genes/proteins,and it was found that they were mainly enriched in the PI3K-Akt signaling pathway,Ras signaling pathway,and MAPK signaling pathway.It is indicated that medical ozone may promote the repair of injured muscle through regulating PI3K/Akt/NF-κB and Ras/MAPK/NF-κB signaling pathway by acting on target genes,such as Syk,FGF16,CSF-1,and MRAS.

Medical therapy and surgical intervention are the two primary approaches for treating myocardial bridge.However,there remains controversy regarding the use of coronary artery bypass grafting(CABG)and myocardial bridge unroofing.Here,we report a case of myocardial infarction following CABG in a patient with a myocardial bridge.The patient was admitted to Lanzhou First Peopie's Hospital with persistent chest pain,chest tightness,and shortness of breath lasting 2 hours.Physical examination revealed no significant abnormalities.Electrocardiography(ECG)indicated extensive anterior wall myocardial infarction.Laboratory findings showed myoglobin levels of 140.1 ng/ml and troponin Ⅰ levels of 2.59 ng/ml,with no other significant abnormalities.The initial diagnosis was acute extensive anterior wall myocardial infarction.Emergency coronary angiography revealed a myocardial bridge in the mid-segment of the left anterior descending artery(LAD).Emergency CABG using the left internal mammary artery to the LAD was performed,leading to symptomatic improvement,and the patient was discharged in stable condition.However,the patient experienced a recurrent myocardial infarction seven years post-surgery and received secondary preventive medical therapy.The patient is currently under ongoing follow-up care.CABG is an effective treatment for myocardial bridge.However,based on the case reported in this study,we recommend careful evaluation of whether a patient may benefit from CABG.

Objective:To observe any effect of regular aerobic exercise on cardiac remodeling in spontaneously hypertensive rats (SHR) and explore the mechanism of endothelial nitric oxide synthase (eNOS) uncoupling.Methods:Thirty 6-week-old healthy male SHR were divided into a sedentary group and an exercise group, each of 15. Another ten age- and sex-matched Wistar-Kyoto rats were set as a normal control group. The animals in the normal control and sedentary groups were fed quietly in their cages, and those in the exercise group performed moderate intensity treadmill exercise for 8 weeks (5 times per week). Forty-eight hours after the last training, echocardiography was applied to document cardiac structure and function in both groups. Wheat germ agglutinin staining and Picrosirius Red staining were used to obtain the cardiomyocyte cross sectional areas (CSAs) and interstitial collagen volume fractions (CVFs) of all of the mice. The rates of cardiomyocyte apoptosis were measured using TUNEL staining, and myocardial tetrahydrobiopterin (BH4) content was measured using high-performance liquid chromatography. Total eNOS, eNOS dimer and eNOS monomer protein expression in the myocardia were detected using western blotting.Results:Compared with the normal control group, the left ventricular wall thickness (LVWT), myocardial CSA, CVF, apoptosis of cardiomyocytes and eNOS monomer levels were significantly higher in the sedentary group, on average. But the end-diastolic diameter (LVEDD) and ejection fraction (LVEF) of the left ventricle and the levels of eNOS dimer and myocardial BH4 and the eNOS dimer/monomer ratio tended to be lower. Comparing the exercise group with the sedentary group, the average LVEDD, LVEF, eNOS dimer, eNOS dimer/monomer ratio and myocardial BH4 content were significantly higher in the exercise group, but the myocardial CVF, cardiomyocyte apoptosis and eNOS monomer levels were significantly lower. LVWT and CSA were not significantly different. There were no significant differences in the total eNOS protein levels among the three groups.Conclusion:Regular aerobic exercise might improve cardiac remodeling in cases of spontaneous hypertension regulating eNOS uncoupling, at least in rats.

Objective:To explore the effect of TP53 mutation variant allele frequency(VAF)on the prognosis of diffuse large B-cell lymphoma(DLBCL)patients.Methods:This study included 155 patients with DLBCL who were first diagnosed in the People's Hospital of Xinjiang Uygur Autonomous Region from March 2009 to March 2022. Complete clinical data and paraffin-embedded tumor tissue samples were obtained,and DNA was extracted from tumor tissues.The gene mutation profile of DLBCL patients was detected and analyzed by second-generation sequencing technology.Kaplan-Meier method was used to analyze the mutation status of TP53 gene and the relationship between mutation VAF and OS.Cox regression univariate and multivariate analysis was use to analyze the independent factors affecting OS.A nornogram model for predicting 1,3,and 5 years OS in DLBCL patients were established to evaluated the performance of the model based on C-index and calibration curves.Results:The average value of TP53 mutation VAF in male DLBCL patients was significantly higher than that in female patients (P<0.05 ).Patients with TP53 mutantion had shorter OS than those with wild-type patients (P=0.030).The optimal VAF threshold for TP53 mutation based on OS stratification was 33.61％(P<0.001),and patients with TP53 mutation VAF≥34％had shorter OS than those with TP53 mutation VAF<34％and wild-type patients (P<0.001).Multivariate Cox analysis showed that TP53 mutation VAF≥34％ was an independent poor predictor of OS (HR=4.05,P<0.001),and IPI score ≥3 was an independent predictor of OS poor (HR=2.27,P=0.008).In combination with factors with independent prognostic significance obtained from multi-factor analysis,we constructed a nomogram model for predicting 1-year,3-year,5-year OS in DLBCL patients.The results showed that the C index of TP53-mutated VAF combined with IPI model was 0.743,which predicted the value of 1-year,3-year,and 5-year OS in DLBCL patients.Calibration curves show that the model has good agreement between predicted and actual survival of DLBCL patients at 1-year,3-year,and 5-year. Conclusion:TP53 mutant VAF has prognostic value in DLBCL patients,and TP53 mutant VAF≥34％ is an independent risk factor for OS in DLBCL patients.The prognosis model of TP53 mutation VAF combined with IPI nomogram constructed in this study has good predictive performance for DLBCL patients.

Objective:To study the quality and stability of commercial ready-to-use tryptone soya agar(TSA)after storing at 2-25 ℃ for different storage duration under dark condition in order to discuss a determination method of validity period for medium.Methods:Three consecutive batches of ready-to-use TSA medium from two manufac-turers were selected and stored at 2-25 ℃ under dark conditions for 30,90 and 180 days,respectively.The appearance,pH,medium suitability and sterility of the medium were tested.Results:The results of appearance,pH,suitability and sterility of TSA medium from two manufacturers for each batch under different storage duration all met the requirements of the Chinese Pharmacopoeia 2020 Volume IV on the quality control of medium.Conclusion:The TSA medium from two manufacturers all met the requirements when stored for 180 days at 2-25 ℃ under dark condition,indicating that the validity period of TSA medium from two manufacturers can reach 180 days.

Objective:To explore the effect of cathepsin L (CTSL) inhibitor on apoptosis of retinal pigment epithelial (RPE) cells and mitochondrial oxidative stress.Methods:RPE cells were cultured in vitro and divided into control group, hydrogen peroxide (H 2O 2) group, and H 2O 2+CTSL inhibitor group. The cells of H 2O 2 group and H 2O 2+CTSL inhibitor group were incubated in the medium containing 400 μmol/L H 2O 2 for 24 hours and 10 μmol/L CTSL inhibitor was added in H 2O 2+CTSL inhibitor group at the same time. The cells of normal group were routinely cultured cells. The follow-up experiment was carried out 24 hours after modeling. The rate of apoptosis was detected by flow cytometry. The expression of CTSL was detected by immunofluorescence staining, Western blot and real time-polymerase chain reaction. The level of mitochondrial super oxide was detected by MitoSOX fluorescent probe, and the mitochondrial structure was observed after MitoTracker staining, the average area, form factors, and branch of mitochondria were quantitatively analyzed. The two groups were compared using two-tailed Student t test, while numerous groups were compared using one-way ANOVA. Results:Compared with control group, the rate of apoptosis in H 2O 2 group was significantly higher ( t=3.307, P=0.029 7), the expression level of CTSL was significantly increased ( t=19.950, 6.916, 14.220; P<0.05). Compared with H 2O 2 group, the expression level of CTSL, the rate of apoptosis and the mitochondrial ROS level in H 2O 2+CTSL inhibitor group were significantly lower ( t=11.940, 4.718, 16.680; P<0.05). The mitochondria of H 2O 2+CTSL inhibitor group were elongated, oval-shaped or rod-shaped, while the mitochondria of H 2O 2 group lost their continuous contour shape and complete structure. The differences of the average area, form factors, and brach of mitochondria among 4 groups were statistically significant ( F=251.700, 34.010, 60.500; P<0.000 1). Conclusions:H 2O 2 can significantly induce apoptosis in RPE cells and increase CTSL expression. CTSL inhibitor can inhibit the H 2O 2-induced apoptosis of RPE cells, lower the mitochondrial super oxide level, and successfully repair the mitochondrial structure.

Objective To investigate the risk factors of hip osteoarthritis(HOA)after hip arthroscopy in patients with femoro-acetabular impingement(FAI)syndrome,and to reduce and prevent HOA.Methods From September 2018 to Septem-ber 2020,106 patients with FAI underwent hip arthroscopy,including 40 males and 66 females,aged from 20 to 55 years old with an average age of(33.05±10.19)years old.The mechanism of injury included 51 cases for sports injury,36 for traffic ac-cidents,and 19 for blunt object injury.The duration of the disease ranged from 5 to 19 days with an average of(12.02±3.69)days.All patients were followed up for 18 months.Patients were divided into HOA group(23 cases)and non-HOA group(83 cases)according to the occurrence of HOA.Multivariate Logistic regression was used to analyze the risk factors of HOA after hip arthroscopy in FAI patients.Results By univariate analysis,aged from 50 to 70 years old,female,body mass index(BMI)＞30 kg·m-2,physical labor,cam type,postoperative infection,last follow-up hip degree of motion(range of motion,ROM)(flex-ion,abduction,adduction,internal rotation)and T?nnis grade 1 and above of the HOA group were higher than those of the non-HOA group(P＜0.05),and the relative appendicular skeletal muscle index(RASM)was lower than that of non-HOA group(P＜0.05).By multiple Logistic regression analysis,cam type,BMI＞30 kg·m-2,last follow-up hip internal rotation ROM and T?nnis grade 1 were risk factors for HOA after hip arthroscopy in FAI patients(P＜0.05).Conclusion FAI classification,body mass index,hip ROM and T?nnis grade are all related to HOA after hip arthroscopy in FAI patients.Follow-up and intervention should be strengthened in high-risk FAI patients to reduce the occurrence of HOA.