Objective To investigate optimal method of establishing hypoxia/reoxygenation(H/R) model of H9c2 cell by using hypoxia/anoxic workstation under different conditions in hypoxia.Methods H9c2 cell was placed into hypoxia/anoxic workstation and simultaneously cultured with complete medium, glucose-free DMEM and acidic hypoxic solution for 1,2,4,6 and 8 h respectively, and then reoxygenated with complete medium for 1 h in normoxic incubator.The level of ROS was measured by flow cytometry.The cell viability was detected by MTT assay.The cellular morphology was observed by inverted microscope.Results With the extension of cell hypoxia time, there were no significant differences in the ROS level and cell viability in complete medium-and glucose-free DMEM-treated H/R groups compared with control group(P<0.05).There was no obvious morphologic change observed with inverted microscope, either.Nevertheless, when H9c2 cells were treated with acidic hypoxic solution in hypoxia, the ROS level continuously increased and the cell viability decreased with the extension of cell hypoxia time ( P<0.01 ).Since H:1 h/R:1 h, some of the cells shrunk and a few necrotic cells floated in the media under the inverted microscope , and the damage was aggravated with the extension of hypoxia time.After the cells were exposed in hypoxia for 8 h, they wrinkled to be round and a large number of floating necrotic cells were observed.When the cells were reoxygenated for 1 h, the cytomembrane was not smooth and there were still a few necrotic cells floating in culture dish .Conclusion The H9c2 cell H/R model with good repeatability can be established successfully by using hypoxia/anoxic workstation combining with acidic hypoxic solution.

Objective To study the effect of Shaoyaozhitong Mixture on stageⅢ-Ⅳendometriosis, and explore its mechanism. Methods Ninty-five cases of stageⅢ-Ⅳ endometriosis were randomly divided into three groups. The traditional Chinese medicine (TCM) group (31 cases) was treated with Shaoyaozhitong Mixture, the GnRHa group (31 cases) was treated with Triptorelin Acetate for Injection, while the expectant group (33 cases) received no medications. The changes of CA125, TNF-α and VEGF levels, visual pain score and SF-36 score, total effective rate, pregnancy and recurrence were compared. Results There were significant increases in CA125, TNF-α and VEGF levels, visual analogue pain score and SF-36 score in all the three groups after treatment (P<0.05). The decline of CA125, TNF-αand VEGF levels and visual analogue pain score in TCM group and GnRHa group was significantly faster and greater than that of the expectant group (P<0.05). The total effective rates of TCM group was 83.9%(26/31), GnRHa group was 87.1%(27/31), and expectant group was 60.6%(20/33), TCM group and GnRHa group were superior to the expectant group, respectively (P<0.05). The pregnancy rate was higher and recurrence rate was lower in TCM group and GnRHa group than that of expectant group, but there was no statistical difference (P>0.05). Conclusion Shaoyaozhitong Mixture is effective in treating endometriosis by inhibiting the growth of ectopic endometrium.

By studying the behaviors of surfing in the Internet of the readers in our electronic reading room,we find a majority of readers are for amusement,only a minority of readers are to study and search data.Aiming at these problems,we put forward a series of measures and management countermeasures that guide readers to make good use of the electronic reading room and give some suggestions for our future work.

Aim To investigate the effect of N-n-butyl haloperidol iodide(F2) on mitochondria-dependent apoptotic pathway of H9c2 cardiac myocytes during hypoxia/reoxygenation(H/R) injury.Methods The H/R models of H9c2 cardiac myocytes were established.The H9c2 cardiac myocytes were randomly divided into five groups: control group(C group), hypoxia/reoxygenation group(H/R group), F2 low concentration group(L), F2 medium concentration group(M), F2 high concentration group(H).Apoptotic rate was evaluated by flow cytometry(FCM).The levels of Cyto C, Bcl-2, Bax were observed by Western blot.Caspase-3 activity was measured with colorimetry.Results Compared with H/R group, F2 low, medium and high concentrations group could significantly decrease apoptosis rate and increase the ratio of Bcl-2 to Bax proteins and inhibit the release of Cyto C into the cytosolic fraction, and decrease caspase-3 activity.Conclusion F2 can protect H9c2 cardiac myocytes against H/R-induced injury through interfering in mitochondria-dependent pathway.

Objective:To investigate the correlation of mitochondrial respiration function and oxidative phosphorylation of peripheral blood mononuclear cells(PBMCs)with grip strength and muscle mass in the elderly, and to identify potential biomarkers for the diagnosis of sarcopenia.Methods:A total of 65 patients admitted to our hospital from June 2019 to August 2020 were enrolled in this study.PBMCs were extracted from subjects.Mitochondrial oxidative respiration function was assessed via the Seahorse XF24 analyzer.Grip strength was measured using a hydraulic dynamometer, and appendicular skeletal muscle mass(ASM)was estimated by dual-energy X-ray absorptiometry(DXA). Multivariate analysis was conducted by using partial correlation analysis and multiple linear regression, in order to evaluate the correlation of mitochondrial oxidative respiration function with grip strength and ASM.Results:After adjustment for gender and body mass index(BMI), partial correlation analysis showed that grip strength and ASM had a negative correlation with age( r=-0.537 and -0.390, both P<0.001); and basal respiration, maximal respiration, ATP production and spare respiratory capacity of mitochondria in PBMCs were negatively correlated with age( r=-0.558, -0.614, -0.526 and -0.582, all P<0.001), whereas grip strength and ASM were positively correlated with basal respiration, maximal respiration, ATP production, spare respiratory capacity and proton leak of mitochondria in PBMCs(grip strength: r=0.414, 0.451, 0.362, 0.420 and 0.425, P=0.002, 0.001, 0.008, 0.002 and 0.002; ASM: r=0.319, 0.368, 0.299, 0.352 and 0.279, P=0.019, 0.006, 0.028, 0.009 and 0.041). Multiple linear regression analysis showed that grip strength and ASM were positively correlated with basal respiration, maximal respiration, ATP production, spare respiratory capacity and proton leak of mitochondria in PBMCs(grip: β=0.503, 0.548, 0.452, 0.519 and 0.532, t=3.248, 3.604, 2.774, 3.301 and 3.350, P=0.002, 0.001, 0.008, 0.002 and 0.002; ASM: β=0.302, 0.355, 0.289, 0.346 and 0.271, t=2.427, 2.856, 2.263, 2.716 and 2.091, P=0.019, 0.006, 0.028, 0.009 and 0.041). Age was negatively correlated with basal respiration, maximal respiration, ATP production and spare respiratory capacity of mitochondria in PBMCs( β=-0.581, -0.654, -0.558 and -0.640, t=-4.285, -5.157, -3.938 and -4.863, all P<0.001). Multiple linear regression analysis showed that ASM and grip strength had no significant correlation with basal respiration, maximal respiration, ATP production, spare respiratory capacity or proton leak of mitochondria in PBMCs. Conclusions:Age-related mitochondrial oxidative respiration in PBMCs can reflect changes in muscle strength and muscle mass and, combined with grip strength and ASM, may be considered as a biomarker for the evaluation of sarcopenia in the elderly.

With the development of the economy and technological progress,more and more animal-derived mesh products are being utilized in the medical field for tissue and organ repair and replacement.Owing to the complexity of their structure and production process,these animal-derived meshes still face several challenges in practical applications,such as insufficient mechanical strength,rapid degradation rates,and the detection of harmful leachable substances.Among these challenges,the production process is a key factor affecting product quality.This paper reviews the key aspects of the production process and quality control for animal-derived meshes in China,offering new insights for the quality control and regulatory oversight of such products.

Mitochondrial dynamics regulates mitochondrial morphology and functions.Imbalance in mitochondrial dynamics with aging leads to mitochondrial dysfunction, accelerates the aging process and is closely related to the occurrence and progression of age-related diseases.However, the specifics of the relationship between aging and altered mitochondrial dynamics are still not fully understood.Here, we review the link between mitochondrial dynamics and aging, and discuss mechanisms underlying age-related diseases associated with altered mitochondrial dynamics, aiming to identify novel therapeutic targets and strategies for the management of age-related diseases.

Objective To explore the activation of tumor necrosis factor-α(TNF-α)signaling pathway and the expressions of the associated inflammatory factors in NPHP1-defective renal tubular epithelial cells.Methods A human proximal renal tubular cell(HK2)model of lentivirus-mediated NPHP1 knockdown(NPHP1KD)was constructed,and the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors CXCL5,CCL20,IL-1β,IL-6 and MCP-1 were detected using RT-qPCR,Western blotting or enzyme-linked immunosorbent assay.A small interfering RNA(siRNA)was transfected in wild-type and NPHP1KDHK2 cells,and the changes in the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors were examined.Results NPHP1KDHK2 cells showed significantly increased mRNA expressions of TNF-α,C/EBPβ,CXCL5,IL-1β,and IL-6(P<0.05),protein expressions of phospho-p38 and C/EBPβ(P<0.05),and IL-6 level in the culture supernatant(P<0.05),and these changes were significantly blocked by transfection of cells with siRNA-C/EBPβ(P<0.05).Conclusion TNF-α signaling pathway is activated and its associated inflammatory factors are upregulated in NPHP1KDHK2 cells,and C/EBPβ may serve as a key transcription factor to mediate these changes.

Objective To explore the activation of tumor necrosis factor-α(TNF-α)signaling pathway and the expressions of the associated inflammatory factors in NPHP1-defective renal tubular epithelial cells.Methods A human proximal renal tubular cell(HK2)model of lentivirus-mediated NPHP1 knockdown(NPHP1KD)was constructed,and the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors CXCL5,CCL20,IL-1β,IL-6 and MCP-1 were detected using RT-qPCR,Western blotting or enzyme-linked immunosorbent assay.A small interfering RNA(siRNA)was transfected in wild-type and NPHP1KDHK2 cells,and the changes in the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors were examined.Results NPHP1KDHK2 cells showed significantly increased mRNA expressions of TNF-α,C/EBPβ,CXCL5,IL-1β,and IL-6(P<0.05),protein expressions of phospho-p38 and C/EBPβ(P<0.05),and IL-6 level in the culture supernatant(P<0.05),and these changes were significantly blocked by transfection of cells with siRNA-C/EBPβ(P<0.05).Conclusion TNF-α signaling pathway is activated and its associated inflammatory factors are upregulated in NPHP1KDHK2 cells,and C/EBPβ may serve as a key transcription factor to mediate these changes.

AIM:To explore the effect of polyphenolic compound 3,5-dihydroxy-4-methoxybenzyl alcohol(DHMBA)on hypoxia/reoxygenation(H/R)injury of human umbilical vein endothelial cells(EA.hy926 cells)and its po-tential mechanisms.METHODS:To construct an H/R model,the EA.hy926 cells were cultured in an acidic hypoxia buffer while in an anaerobic workstation.The cells were divided into control,H/R,H/R+different doses of DHMBA,H/R+edaravone(antioxidant)and H/R+reactive oxygen species(ROS)inducer oligomycin A+DHMBA groups.Cell viability was measured by CCK-8 assay,and tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels in cells were mea-sured by ELISA.Phosphorylation of endothelial nitric oxide synthase(eNOS)and nuclear factor-κB(NF-κB)p65 were measured by Western blot.Intracellular NO levels were determined by laser confocal microscopy.Glutathione(GSH)/glu-tathione disulfide(GSSG)oxidation balance was determined by the dinitrobenzoic acid chromogenic method.Intracellular ROS levels were measured by flow cytometry.Lactate dehydrogenase(LDH)leakage was determined using nitro blue tet-razolium staining.Scratch assays were performed to assess cell migration.RESULTS:DHMBA exhibited no significant cytotoxicity between 125 and 1 000 μmol/L.In H/R-injured human umbilical vein endothelial cells,DHMBA improved cell survival,inhibited phosphorylation of NF-κB p65,reduced the content of TNF-α and IL-6,and increased phosphory-lation of eNOS and NO levels.DHMBA also suppressed ROS overload and restored the ratio between GSH and oxidized GSH,decreased in LDH leakage and increased cell migration in H/R-injured human umbilical vein endothelial cells.CONCLUSION:DHMBA can alleviate H/R-induced oxidative stress,inflammation,cellular damage,and dysfunction,which are associated with the ability of DHMBA to inhibit ROS production in human umbilical vein endothelial cells.