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1.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 35-44, 2024.
Article in Chinese | WPRIM | ID: wpr-1007272

ABSTRACT

ObjectiveTo investigate the effect of circSLC8A1_005 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe effect of adenovirus-mediated overexpression of circSLC8A1_005 on the expression of fibrosis-related genes, collagen type I alpha 1 chain (Col1a1), collagen type Ⅲ alpha 1 chain (Col3a1) and smooth muscle actin alpha 2 (Acta2), in mouse cardiac fibroblasts (mCFs) were detected. The proliferation and migration activities of mCFs were detected by EdU and wound-healing assay, respectively. Dual luciferase reporter gene assay was performed to detect the activity of potential internal ribozyme entry site (IRES) in circSLC8A1_005. CircSLC8A1_005-translated protein, SLC8A1-605aa, and its intracellular distribution was identified by Western blot assay. The effect of SLC8A1-605aa protein on transcription activity of Sod2 gene was detected by the dual luciferase reporter gene assay. RNA binding protein immunoprecipitation (RIP) was utilized to verify the interaction between SLC8A1-605aa and superoxide dismutase 2 (Sod2) mRNA. Actinomycin D treatment was used to detect the effect of SLC8A1-605aa on Sod2 mRNA stability in mCFs. ResultsAn efficient adenovirus-mediated overexpression of circSLC8A1_005 was achieved in mCFs. The enforced expression of circSLC8A1_005 suppressed proliferation and migration of mCFs, and inhibited the expression of fibrosis-related genes in mCFs. The dual luciferase reporter gene assay revealed the activities of 2 IRES in circSLC8A1_005. Results of Western blot assay showed that circSLC8A1_005 could translate protein SLC8A1-605aa with the prospected molecular weight of 70 ku, which is predominantly distributed in the nucleus. Overexpression of the circSLC8A1_005 and SLC8A1-605aa could consistently inhibit the fibrotic phenotype of mCFs. SLC8A1-605aa could up-regulate superoxide dismutase 2 (Sod2) expression, but not at the transcriptional level. RIP assay indicated that SLC8A1-605aa could specifically interact with Sod2 mRNA, and the results of actinomycin D assay showed that SLC8A1-605aa could enhance the stability of Sod2 mRNA in mCFs. ConclusionCircSLC8A1_005 inhibits the fibrotic phenotype of cardiac fibroblasts via translating SLC8A1-605aa protein, and SLC8A1-605aa may be a potential target for the treatment of myocardial fibrosis.

2.
Acta Pharmaceutica Sinica ; (12): 25-34, 2024.
Article in Chinese | WPRIM | ID: wpr-1005435

ABSTRACT

Understanding the research methods for drug protein targets is crucial for the development of new drugs, clinical applications of drugs, drug mechanisms, and the pathogenesis of diseases. Cellular thermal shift assay (CETSA), a target research method without modification, has been widely used since its development. Now, there are various CETSA-based technology combinations, such as mass spectrometry-based cellular thermal shift assay (MS-CETSA), isothermal dose response-cellular thermal shift assay (ITDR-CETSA), amplified luminescent proximity homogeneous assay-cellular thermal shift assay (Alpha-CETSA), etc., which combine their respective advantages and further expand the application scope of CETSA. These technologies are suitable for the entire drug development chain, from drug screening to monitoring the target binding and off-target toxicity of drugs in patients. Based on the author's research experience, this paper reviews the principles of CETSA and related binding technologies, their application in target discovery, and the progress of data processing and analysis in recent years, aiming to provide reference and reference for the further application of CETSA.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 208-215, 2024.
Article in Chinese | WPRIM | ID: wpr-999178

ABSTRACT

ObjectiveThis study explored the application of Yiqi Zengmian prescription as a vaccine adjuvant, aiming to provide a new scheme for the prevention and control of corona virus disease 2019(COVID-19) with traditional Chinese medicine (TCM). By analyzing the compatibility and efficacy, this paper examines the compatibility effect of Yiqi Zengmian prescription, which is modified from the classic tonifying agent Si Junzitang, as a vaccine adjuvant. MethodUsing the Database of Ancient Classical Prescriptions, this paper analyzed the composition of Yiqi Zengmian prescription and probed into the theoretical basis for the compatibility of this prescription from the properties, medicine combination, and efficacy. Furthermore, the compatibility effect of this prescription with vaccines was analyzed. ResultAs a TCM prescription, Yiqi Zengmian prescription focuses on the lung and spleen and enhances the Qi in the two organs. The lung governs Qi movement. The body breathes fresh air through the lungs and exchanges the turbid gas in the lungs, and the gas circulates alternately in the lungs to ensure the normal breathing of the human body. The spleen governing transportation and transformation is the hub for Qi movement, and Qi is the embodiment of metabolic function. By regulating qi movement and enhancing the functions of Qi and blood, Yiqi Zengmian prescription can enhance the immunogenicity of the vaccine, which provides a theoretical basis for enhancing the immune effects of vaccines. ConclusionYiqi Zengmian prescription has the effects of replenishing Qi and invigorating spleen, regulating Qi and drying dampness, and enhancing immunity. The in-depth analysis of the TCM theory of Yiqi Zengmian prescription as a vaccine adjuvant and the results of clinical and laboratory studies suggest that Yiqi Zengmian prescription may enhance the induction of immune response after vaccination and maintain the immune memory. However, the mechanism of Yiqi Zengmian prescription in regulating the complex immune network remains to be elucidated.

4.
Acta Pharmaceutica Sinica ; (12): 418-423, 2024.
Article in Chinese | WPRIM | ID: wpr-1016651

ABSTRACT

Twelve compounds were isolated from the ethyl acetate fraction of the 80% aqueous ethanol extract of the roots and stems of Dalbergia rimosa Roxb. by silica gel, MCI, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Their structures were identified by spectral analysis such as UV, IR, MS, 1D/2D NMR and by comparison with literature information as dalbergiquinol A (1), dalbergiquinol B (2), R-(-)-3′-hydroxy-2,4,5-trimethoxydalbergiquinol (3), neokhriol A (4), mucronulatol (5), (3R)-7,2′,3′-trihydroxy-4′-methoxy-isoflavane (6), isomucronulatol (7), (3S)-violanone (8), 3′-O-methylviolanone (9), eryvarin M (10), (±)-α,3,4,2′,4′-pentahydroxydihydrochalcone (11) and (-)-butin (12). Compound 1 and 2 are new compounds, and compounds 3-12 were isolated from this plant for the first time. Compounds 1, 2, 4, 6, 8, 11, 12 showed good scavenging effect on DPPH free radical.

5.
Acta Pharmaceutica Sinica ; (12): 621-632, 2024.
Article in Chinese | WPRIM | ID: wpr-1016624

ABSTRACT

Idiosyncratic drug-induced liver injury (IDILI) has long posed a challenging and pivotal concern in pharmaceutical research. The complex composition of traditional Chinese medicine (TCM) has introduced a bottleneck in current research, hindering the elucidation of the component basis associated with IDILI in TCM. Using Epimedii Folium (EF) and Psoraleae Fructus (PF) as illustrative examples, this study endeavors to establish an in vitro evaluation model, providing a high-throughput and preliminary assessment method for screening components related to TCM-induced IDILI. A TNF-α-mediated HepG2 susceptible model was first established in this study, with the focus on the index components present in EF and PF. The release of lactate dehydrogenase (LDH) in the cell supernatant served as the detection index. A concentration-toxicity response curve was constructed, and the hepatotoxic components of EF and PF were identified utilizing the synergistic toxicity index. The LDH results unveiled the hepatotoxic effects of bavachin, backuchiol, isobavachin, neobavaisoflavone, psoralidin, isobavachalcone, icarisid I, and icarisid II on both normal and susceptible cells, categorizing these 8 components as both direct hepatotoxicity components and idiosyncratic hepatotoxicity components. Bavachin and neobavaisoflavone exhibited no hepatotoxicity on normal cells but demonstrated significant effects on susceptible cells, designating them as potential idiosyncratic susceptible hepatotoxicity components. The study further delineated that 10 EF components and 3 PF components were direct immune-promoting hepatotoxicity components. Additionally, 14 idiosyncratic immune-promoting hepatotoxicity components were identified, encompassing 10 EF components and 4 PF components, with neobavaisoflavone, bavachinin, and isobavachin being potential idiosyncratic susceptible immune-promoting hepatotoxicity components. Synergistic toxicity index results indicated that 13 idiosyncratic immune-promoting hepatotoxicity components (except anhydroicaritin) combined with bavachin demonstrated synergistic hepatotoxicity on susceptible cells. Notably, 3 idiosyncratic susceptible immune-promoting hepatotoxicity components combined with bavachin exhibited synergistic hepatotoxicity, with neobavaisoflavone displaying the highest synergistic toxicity index and bavachinin the lowest. In summary, this methodology successfully screens hepatotoxic and immune-promoting hepatotoxic components in EF and PF, distinguishing the types of components inducing hepatotoxicity, evaluating the hepatotoxicity degree of each component, and elucidating the synergistic relationships among them. Importantly, these findings align with the characteristics of IDILI. The method provides an effective model tool for the fundamental research of TCM-related IDILI components.

6.
Acta Anatomica Sinica ; (6): 55-61, 2024.
Article in Chinese | WPRIM | ID: wpr-1015142

ABSTRACT

Objective To investigate the relationship between nuclear factor(NF)-κB signaling pathway and gender differences in alcoholic liver fibrosis. Methods C57BL/6 N mice at 7-8 weeks of age were randomly divided into: male normal group, male model group, female normal group and female model group of 20 mice each. The normal group was fed with control liquid diet for 8 weeks, and the model group was fed with alcoholic liquid diet for 8 weeks combined with 31.5% ethanol gavage (5g/kg twice a week) to establish an alcoholic liver fibrosis model. The mice were executed at the end of 8 weekends, and the alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity, estradiol (E

7.
Cancer Research on Prevention and Treatment ; (12): 127-133, 2024.
Article in Chinese | WPRIM | ID: wpr-1011510

ABSTRACT

Lung cancer is one of the most common cancers worldwide, and its mortality rate remains high. In addition to conventional surgery, radiotherapy, and chemotherapy, immunotherapy methods have been developed and used in recent years for the treatment of non-small cell lung cancer (NSCLC). However, only a small number of patients with NSCLC can benefit from immunotherapy strategies, and some patients even have hyperprogression after receiving immunotherapy. Therefore, precision immunotherapy requires effective biomarkers to guide it. In this paper, tissue samples, blood samples, intestinal microbiota, and other biomarkers are reviewed according to different sample sources. Blood samples, including TCR immune repertoire, Tregs cells, cytokines, lactate dehydrogenase, and other markers, are summarized and analyzed to provide reference for clinicians' diagnosis and treatment decisions.

8.
Chinese journal of integrative medicine ; (12): 243-250, 2024.
Article in English | WPRIM | ID: wpr-1010328

ABSTRACT

OBJECTIVE@#To investigate the effects of Danmu Extract Syrup (DMS) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the mechanism.@*METHODS@#Seventy-two male Balb/C mice were randomly divided into 6 groups according to a random number table (n=12), including control (normal saline), LPS (5 mg/kg), LPS+DMS 2.5 mL/kg, LPS+DMS 5 mL/kg, LPS+DMS 10 mL/kg, and LPS+Dexamethasone (DXM, 5 mg/kg) groups. After pretreatment with DMS and DXM, the ALI mice model was induced by LPS, and the bronchoalveolar lavage fluid (BALF) were collected to determine protein concentration, cell counts and inflammatory cytokines. The lung tissues of mice were stained with hematoxylin-eosin, and the wet/dry weight ratio (W/D) of lung tissue was calculated. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-1 β in BALF of mice were detected by enzyme linked immunosorbent assay. The expression levels of Claudin-5, vascular endothelial (VE)-cadherin, vascular endothelial growth factor (VEGF), phospho-protein kinase B (p-Akt) and Akt were detected by Western blot analysis.@*RESULTS@#DMS pre-treatment significantly ameliorated lung histopathological changes. Compared with the LPS group, the W/D ratio and protein contents in BALF were obviously reduced after DMS pretreatment (P<0.05 or P<0.01). The number of cells in BALF and myeloperoxidase (MPO) activity decreased significantly after DMS pretreatment (P<0.05 or P<0.01). DMS pre-treatment decreased the levels of TNF-α, IL-6 and IL-1 β (P<0.01). Meanwhile, DMS activated the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway and reversed the expressions of Claudin-5, VE-cadherin and VEGF (P<0.01).@*CONCLUSIONS@#DMS attenuated LPS-induced ALI in mice through repairing endothelial barrier. It might be a potential therapeutic drug for LPS-induced lung injury.


Subject(s)
Mice , Male , Animals , Proto-Oncogene Proteins c-akt/metabolism , Lipopolysaccharides , Phosphatidylinositol 3-Kinases/metabolism , Interleukin-1beta/metabolism , Vascular Endothelial Growth Factor A/metabolism , Tumor Necrosis Factor-alpha/metabolism , Claudin-5/metabolism , Acute Lung Injury/chemically induced , Lung/pathology , Interleukin-6/metabolism , Drugs, Chinese Herbal
9.
International Eye Science ; (12): 1001-1006, 2023.
Article in Chinese | WPRIM | ID: wpr-973794

ABSTRACT

AIM:To explore the use of attention mechanism and Pix2Pix generative adversarial network to predict the postoperative corneal topography of age-related cataract patients undergone femtosecond laser arcuate keratotomy.METHODS:In this retrospective case series study, the 210 preoperative and postoperative corneal topographies from 87 age-related cataract patients(105 eyes)undergoing femtosecond laser arcuate keratotomy at Shanxi Eye Hospital between March 2018 and March 2020 were selected and divided into a training set(180)and a test set(30)for model training and testing. The peak signal-to-noise ratio(PSNR), structural similarity(SSIM)and Alpins astigmatism vector analysis were used to compare the accuracy of postoperative corneal topography prediction under different attention mechanisms.RESULTS:The model based on attention mechanism and Pix2Pix network can predict postoperative corneal topography, among which the model based on Self-Attention mechanism has the best prediction effect, with PSNR and SSIM reaching 16.048 and 0.7661, respectively. There were no statistically significant differences in the difference vector, difference vector axis position, surgically induced astigmatism, and correction index between real and generated corneal topography on the 3mm and 5mm rings(all P&#x003E;0.05).CONCLUSION:Based on the Self-Attention mechanism and Pix2Pix network, the postoperative corneal topography can be well predicted, which can provide reference for the surgical planning and postoperative effects of ophthalmic clinicians.

10.
Journal of Public Health and Preventive Medicine ; (6): 124-127, 2023.
Article in Chinese | WPRIM | ID: wpr-973374

ABSTRACT

Hypertension is currently the most common chronic disease in the elderly, and frailty is a non-specific state in which vulnerability of the body increases and anti-stress ability decreases due to physiological reserve function decline. A series of clinical adverse events may occur in elderly people with frailty when there is a small stimulus. Frailty and hypertension often coexist in the elderly. Relevant guidelines for diagnosis and treatment recommend that elderly patients with hypertension should be evaluated for frailty when using antihypertensive drugs. The risks and benefits of antihypertensive treatment should be evaluated according to the patient's physiological reserves, thereby helping to choose personalized antihypertensive treatment, determining appropriate antihypertensive goals, and improving the management of elderly hypertension. Herein, the research progress on hypertension related frailty in the elderly is reviewed to provide reference for personalized treatment of elderly patients with hypertension.

11.
Journal of Clinical Hepatology ; (12): 1191-1196, 2023.
Article in Chinese | WPRIM | ID: wpr-973216

ABSTRACT

Transjugular intrahepatic portosystemic shunt (TIPS) is a safe and effective method for the treatment of portal hypertension complications in patients with decompensated liver cirrhosis. At present, there are many prognostic scoring tools for risk stratification of poor prognosis after TIPS. This article briefly introduces seven prognostic scoring tools commonly used for TIPS and summarizes the clinical research evidence of each scoring tool. The literature review shows that there is currently no sufficient research evidence to determine the optimal prognostic scoring tool after TIPS. Future clinical studies should comprehensively explore the advantages and disadvantages of different scoring tools in predicting short- and long-term adverse prognostic events after TIPS and develop new prognostic scoring tools in combination with new prognostic markers.

12.
China Pharmacy ; (12): 1066-1070, 2023.
Article in Chinese | WPRIM | ID: wpr-972948

ABSTRACT

OBJECTIVE To study the inhibitory effect mechanism of rhynchophylline solid lipid nanoparticles (Rhy-SLN) on the proliferation of airway smooth muscle cells (ASMCs) in asthmatic model mice. METHODS Asthma model was prepared by ovalbumin+calmogastrin sensitization. The primary isolation and culture of ASMCs were performed, and morphological observation and identification were also conducted [when α -smooth muscle actin (α -SMA) appeared red and Desmin appeared green in ASMCs, indicating successful cultivation of ASMCs]. The cells were divided into blank group (ASMCs of normal mice), model group (ASMCs of asthma model mice), Rhy-SLN group (ASMCs of asthma model mice), recombinant suppressors of cytokine signaling 1 (SOCS1) overexpression group (ASMCs of asthma model mice transfected with SOCS1 vector), SOCS1-RNAi group (ASMCs of asthma model mice transfected with SOCS1-RNAi vector) and SB203580 group [p38 mitogen-activated protein kinase (p38 MAPK) inhibitor, ASMCs of asthma model mice]. The cells of each group were added into the corresponding culture medium containing drug (10 μmol/L) or not containing drug for 24 hours. MTT method was used to detect the proliferation of ASMCs in asthmatic mice; Western blot assay was used to detect the protein expressions of α-SMA, interleukin-1β (IL-1β), SOCS1, p38 MAPK and phosphorylated p38 MAPK (p-p38 MAPK) in ASMCs. RESULTS The primary ASMCs of mice varied in shape and size, presenting irregular, spindle and triangular shapes;α-SMA appeared red and Desmin appeared green, indicating successful cultivation of ASMCs. Compared with model group, ASMCs absorbance values and protein expressions of α -SMA, p38 MAPK, and p-p38 MAPK were reduced significantly in Rhy- SLN group, SOCS1 overexpression group and SB203580 E-mail:wangmeng106@163.com group, while protein expression of SOCS1 (except for group) was increased significantly (P<0.05); protein expressions of IL-1β was reduced significantly in ASMCs (P< 0.05). ASMCs absorbance values and protein expressions of α-SMA, SOCS1, p38 MAPK and p-p38 MAPK were increased significantly in SOCS1-RNAi group (P<0.05). CONCLUSIONS Rhy-SLN can inhibit the proliferation of ASMCs, the mechanism of which may be associated with overexpression of SOCS1 and inhibiting the protein expressions of IL-1β and p38 MAPK.

13.
Journal of Clinical Hepatology ; (12): 718-722, 2023.
Article in Chinese | WPRIM | ID: wpr-971919

ABSTRACT

A-kinase anchor protein 12 (AKAP12) is a scaffold protein that improves the specificity and efficiency of spatio-temporal signals by assembling intracellular signal proteins into specific complexes. In recent years, the role of AKAP12 in chronic liver diseases has attracted more and more attention. This article introduces the physiological functions of AKAP12 and reviews the role of AKAP12 in chronic liver diseases, in order to lay a foundation for the use of AKAP12 small molecule as a new therapeutic target for chronic liver diseases.

14.
Journal of Public Health and Preventive Medicine ; (6): 39-43, 2023.
Article in Chinese | WPRIM | ID: wpr-959043

ABSTRACT

Objective To investigate the HPV vaccination status of children aged 9-14 years, parents’ vaccine-related cognition, willingness to vaccinate children and their influencing factors. Methods From January to April 2021, the convenience sampling method was used to collect data based on the online platform and the Department of Pediatrics and Adolescent Gynecology of the Children's Hospital of Zhejiang University School of Medicine. The data were analyzed by descriptive analysis, Chi-square test, and logistic regression. Results A total of 864 questionnaires were collected, and 846 valid questionnaires were obtained after screening, with an effective rate of 97.9%. 13.57% of mothers and 3.09% of girls were vaccinated, and the vaccine awareness rate was 68.43%. 87.22% of parents were willing to bring their children for vaccination. Risk perception (OR=4.79, 95% CI: 2.22-10.35), willingness to vaccinate themselves (OR =29.01, 95% CI: 12.62-66.69), awareness of sex education (OR =3.73, 95% CI: 1.08-12.83) and whether the vaccines were free (P<0.001) were related to whether parents were willing to vaccinate their children. Conclusion Parents of children aged 9-14 have high awareness of HPV and HPV vaccine and are willing to vaccinate their children with HPV vaccine , but the vaccination rate in children is low. Disease perception, willingness to vaccinate, awareness of sex education, and whether vaccines are free are all the factors influencing parents' willingness to vaccinate children.

15.
Journal of Public Health and Preventive Medicine ; (6): 66-67, 2023.
Article in Chinese | WPRIM | ID: wpr-996418

ABSTRACT

Objective To analyze the disease burden status, trends and possible influencing factors of pancreatic cancer in Chengde of Hebei from 2010 to 2020, in order to provide theoretical basis for the preventionof pancreatic cancer. Methods Using the global burden of disease open database, the incidence rate, mortality, years of life lost (YLL), years of life lost with disability (YLD) and disability adjusted life year (DALY) of pancreatic cancer in the region are obtained,average annual percent change (AAPC) was calculated using joinpoint model to test the trend of disease burden change of pancreatic cancer patients from 2010 to 2020. At the same time, the patient characteristics such as hypertension, diabetes and other categorical variables were set as dummy variables, and the risk factors affecting the mortality of patients with pancreatic cancer were analyzed by linear regression. Results In 2010, there were 15 new cases of pancreatic cancer and 13 deaths in Chengde District , Hebei Province. The YLLs caused by pancreatic cancer accounted for 70.67% of DALY. In 2020, there were 160 new cases of pancreatic cancer and 147 deaths in Chengde, Hebei Province. The YLLs caused by pancreatic cancer accounted for 96.02% of DALY. From 2010 to 2020, the incidence of pancreatic cancer increased by 9.79%, and the incidence rate increased by 7.81%, showing an obvious upward trend (APCC =2.20%, P 28.0 and pancreatitis (OR=1.574 , 95% CI: 1.328-3.045) were all risk factors for death of patients with pancreatic cancer (OR>1) . Conclusion From 2010 to 2020, the incidence and mortality of pancreatic cancer among local residents in Chengde of Hebei showed an upward trend, and the disease burden was also increasing year by year. The basic diseases of diabetes and chronic pancreatitis increase the death risk and should be protected.

16.
Cancer Research and Clinic ; (6): 424-428, 2023.
Article in Chinese | WPRIM | ID: wpr-996251

ABSTRACT

Objective:To investigate the application value of CT-guided localization with medical glue or puncture needle before thoracoscopic pulmonary ground-glass nodule resection.Methods:The clinical data of 92 patients who underwent localization before thoracoscopic resection of pulmonary ground-glass nodules in Beijing Chao-Yang Hospital from June 2019 to November 2022 were retrospectively analyzed. According to the preoperative localization method, the patients were divided into puncture needle group (52 cases) and medical glue group (40 cases). The localization success rate, localization time and incidence rate of complications were compared between the two groups.Results:The success rate of localization in both groups was 100%. In puncture needle group, the guide wire of 1 case (1.9%) prolapsed, but it did not affect the surgical resection. The localization time of puncture needle group and medical glue group was (18±6) min and(14±5) min, and there was a statistical difference between the two groups ( t = 3.06, P = 0.003). The incidence of bleeding and pneumothorax in medical glue group was lower than that in puncture needle group [12.5% (5/40) vs. 38.5% (20/52), χ2 = 7.70, P = 0.009; 35.0% (14/40) vs. 71.2% (37/52), χ2 = 11.96, P = 0.001]. The incidence rate of irritating cough in was higher than that in puncture needle group [50.0% (20/40) vs. 11.5% (6/52), χ2 = 16.50, P < 0.05]. Conclusions:CT-guided localization with medical glue or puncture needle can achieve satisfactory results in the localization of pulmonary ground-glass nodule before thoracoscopic resection and help to improve the accuracy of surgical resection. The incidence rates of bleeding and pneumothorax of medical glue localization are lower than those of puncture needle localization.

17.
Chinese Journal of Ocular Fundus Diseases ; (6): 408-414, 2023.
Article in Chinese | WPRIM | ID: wpr-995644

ABSTRACT

Objective:To observe the effect of metformin (Met) on inflammatory bodies and focal death in human retinal microvascular endothelial cells (hRMEC) in diabetes mellitus (DM) microenvironment.Methods:Experimental research was divided into in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 9 healthy C57BL/6J male mice were randomly divided into DM group, normal control group, and DM+Met group, with 3 mice in each group. DM group and DM+Met group mice were induced by streptozotocin to establish DM model, and DM+Met group was given Met 400 mg/ (kg · d) intervention. Eight weeks after modeling, the expression of NLRP3, cleaved-membrane perforating protein D (GSDMD) and cleaved-Caspase-1 in the retina of mice in the normal control group, DM group and DM+Met group were observed by immunohistochemical staining. In vitro cell experiments: hRMEC was divided into conventional culture cell group (N group), advanced glycation end products (AGE) group, and AGE+Met group. Joining the AGE, AGE+Met groups cells were induced by 150 μg/ml of glycation end products, and 2.0 mmol/L Met was added to the AGE+Met group. Pyroptosis was detected by flow cytometry; 2' ,7'-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe was used to detect the expression of reactive oxygen species (ROS) in cells of each group. Real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the relative mRNA and protein expression levels of NLRP3, cleaved-GSDMD, cleaved-Caspase-1 in each group of cells. Single factor analysis of variance was used for comparison among the three groups.Results:In vivo animal experiments: compared with the DM group, the expression of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in the retina of normal control group and DM+Met group mice was significantly reduced, with significant difference among the 3 groups ( F=43.478, 36.643, 24.464; P<0.01). In vitro cell experiment and flow cytometry showed that the pyroptosis rate of AGE group was significantly higher than that of N group and AGE+Met group ( F=32.598, P<0.01). The DCFH-DA detection results showed that the intracellular ROS levels in the N group and AGE+Met group were significantly lower than those in the AGE group, with the significant difference ( F=47.267, P<0.01). The mRNA ( F=51.563, 32.192, 44.473; P<0.01) and protein levels ( F=63.372, 54.463, 48.412; P<0.01) of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in hRMEC of the AGE+Met group were significantly reduced compared to the N group. Conclusion:Met can down regulate the expression of NLRP3 inflammatory body related factors in hRMEC and inhibit pyroptosis.

18.
Chinese Journal of Ocular Fundus Diseases ; (6): 401-407, 2023.
Article in Chinese | WPRIM | ID: wpr-995643

ABSTRACT

Objective:To observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells.Methods:The experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. Results:In vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance ( t=25.372, 22.419, 25.372; P<0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance ( F=36.821, 38.692, 29.421; P<0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant ( F=39.649, P<0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance ( F=27.472, 22.315, 31.147, 27.472; P<0.05). Conclusion:Over-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.

19.
Chinese Journal of Ocular Fundus Diseases ; (6): 324-329, 2023.
Article in Chinese | WPRIM | ID: wpr-995632

ABSTRACT

Objective:To observe the effect of high expression of polypyrimidine tract-binding protein-associated splicing factor (PSF) on low concentration of 4-hydroxynonenal (4-HNE) induced human retinal microvascular endothelial cells (HRMECs), and explore the possible mechanism.Methods:The HRMECs cultured in vitro were divided into 4-HNE treated group, PSF overexpression group combined with 4-HNE group (PSF+4-HNE group), PSF overexpression+ML385 treatment combined with 4-HNE group (PSF+ML385+4-HNE group), and 4-HNE induced PSF overexpression group with LY294002 pretreatment (LY294002+4-HNE+PSF group). Cell culture medium containing 10 μmmol/L 4-HNE was added into 4-HNE treatment group, PSF+4-HNE group, PSF+ML385+4-HNE group for 12 hours to stimulate oxidative stress. 1.0 μg of pcDNA-PSF eukaryotic expression plasmid were transfected into PSF+4-HNE group and PSF+ML385+4-HNE group to achieve the overexpression of PSF. Also cells were pretreated with ML385 (5 μmol/L) for 48 hours in the PSF+ML385+4-HNE group, meanwhile within the LY294002+4-HNE+PSF group, after pretreatment with LY294002, cells were treated with plasmid transfection and 4-HNE induction. Transwell detects the migration ability of PSF to HRMECs. The effect of PSF on the lumen formation of HRMECs was detected by using Matrigel in vitro three-dimensional molding method. Flow cytometer was used to detect the effect of PSF overexpression on reactive oxygen (ROS) level in HRMECs. Protein immunoblotting was used to detect the relative expression of PSF, nuclear factor E2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1) protein, and phosphoserine threonine protein kinase (pAkt) protein. The comparison between the two groups was performed using a t-test. Results:The number of live cells, migrating cells, and intact lumen formation in the 4-HNE treatment group and the PSF+4-HNE group were 1.70±0.06, 0.80±0.13, 24.00±0.58, 10.00±0.67, and 725.00±5.77, 318.7±12.13, respectively. There were significant differences in the number of live cells, migrating cells, and intact lumen formation between the two groups ( t=12.311, 15.643, 17.346; P<0.001). The results of flow cytometry showed that the ROS levels in the 4-HNE treatment group, PSF+4-HNE group, and PSF+ML385+4-HNE group were 816.70±16.67, 416.70±15.44, and 783.30±17.41, respectively. There were statistically significant differences between the two groups ( t=16.311, 14.833, 18.442; P<0.001). Western blot analysis showed that the relative expression levels of pAkt, Nrf2, and HO-1 proteins in HRMECs in the 4-HNE treatment group, PSF+4-HNE group and LY294002+4-HNE+PSF group were 0.08±0.01, 0.57±0.04, 0.35±0.09, 0.17±0.03, 1.10±0.06, 0.08±0.11 and 0.80±0.14, 2.50±0.07, 0.50±0.05, respectively. Compared with the PSF+4-HNE group, the relative expression of pAkt, Nrf2, and HO-1 proteins in the LY294002+4-HNE+PSF group decreased significantly, with significant differences ( t=17.342, 16.813, 18.794; P<0.001). Conclusion:PSF upregulates the expression of HO-1 by activating the phosphatidylinositol 3 kinase/Akt pathway and inhibits cell proliferation, migration, and lumen formation induced by low concentrations of 4-HNE.

20.
Chinese Journal of Thoracic and Cardiovascular Surgery ; (12): 1-3, 2023.
Article in Chinese | WPRIM | ID: wpr-995521

ABSTRACT

With the change of COVID-19 prevention and control strategy in China, the number of COVID-19 cases has increased significantly recently, which has also brought new challenges to the perioperative risk control of thoracic surgery. This paper puts forward several suggestions, aiming to standardize the preoperative screening and evaluation during the COVID-19 period, strictly grasp the indications and timing of surgery, optimize the medical management process, individualize surgical decision-making, and minimize the risk of COVID-19 infection to surgery.

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