24-dehydrocholesterol reductase ameliorates senescence-related dysfunction of vascular endothelial cells / 中华老年医学杂志

Objective:To investigate the role of 24-dehydrocholesterol reductase(DHCR24)in doxorubicin-induced senescence-related dysfunction of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were induced with 0.05 μM doxorubicin for 48 h to establish a stress-triggered premature senescence model.The lentiviral transfection method was employed to achieve DHCR24 overexpression in HUVECs.Cell senescence was evaluated by β-galactosidase staining and Western blot to detect the expression of the senescence-related molecules cyclin-dependent kinase inhibitor 1A(P21)and nicotinamide adenine dinucleotide dependent histone deacetylase 1(SIRT1).Western blot was performed to detect DHCR24 and endothelial nitric oxide synthase(eNOS)expression during endothelial senescence.DAF-FM DA(an NO fluorescent probe)was used to detect intracellular NO production.Results:In the stress-triggered premature senescence model of HUVECs induced by doxorubicin, the expression of the senescence marker P21 was up-regulated( t=19.44, P<0.01), SIRT1 was down-regulated( t=10.10, P<0.01, and the expression of DHCR24 was down-regulated( t=5.946, P<0.01), compared with the control group.Meanwhile, eNOS and NO expression was inhibited( t=11.26, P<0.01; t=10.83, P<0.01).After DHCR24 overexpression, compared with the control stimulation group, the overexpression stimulation group showed that DHCR24( F=72.10, P<0.01)was up-regulated.DHCR24 overexpression alleviated the doxorubicin-induced decrease in eNOS and NO( F=5.797, P<0.05; F=45.12, P<0.01), compared with the control group. Conclusions:DHCR24 may mitigate doxorubicin-induced senescence-related vascular endothelial dysfunction by modulating the eNOS/NO signaling pathway.

Research progress on the role of cell aging in dry age-related macular degen-eration / 眼科新进展

Dry age-related macular degeneration(AMD)is a degenerative disease affecting the macular region of the retina,and aging changes in retinal and choroidal tissues are an important factor in AMD pathogenesis.Cell aging is an irre-versible state of cell cycle arrest triggered by certain physiological processes or stressful injury,affecting a variety of physi-ological and pathological processes.An increasing number of studies have shown that cell aging plays an essential role in the occurrence and development of AMD.This paper reviews the mechanisms of cell aging and its relationship with dry AMD,aiming to provide new ideas for the treatment of dry AMD.

Research progress on biological mechanism of interferon-inducible protein 16 / 中国药理学通报

Interferon γ-inducible protein 16(IFI16)is one member of human pyrin and HIN domain-containing protein(PYHIN)family(also known as interferon-inducible p200 pro-tein family),which is widely present in human organs and tis-sues,and is involved in cell cycle regulation,senescence and ap-optosis,even in immune reaction.The content and localization of IFI16 may change under different physiological and pathological conditions,and recent studies have revealed that it may play an important role in the development of antiviral,tumor,inflammato-ry diseases and other diseases.In this paper,we review its mechanism and the current status of its research in diseases,with the aim of providing a reference for the in-depth study of IFI16.

Research progress of cell senescence in diabetic foot ulcers / 中华老年医学杂志

Diabetic foot ulcers are common in elderly people with diabetes.Ongoing research is aimed at unraveling the complicated molecular mechanisms underlying diabetic foot ulcers.The role of cellular aging in delayed healing of diabetic foot ulcers has been recognized.Analyzing the relationship between cellular aging and diabetic foot ulcers, examining how cellular aging delays the healing of diabetic foot wounds, and identifying treatment methods for the removal of senescent cells and for slowing down cellular senescence will provide new therapeutic targets and strategies for the development of drugs for diabetic foot ulcers.

Metformin inhibits ultraviolet A-induced photoaging of HaCaT cells by activating the adenosine monophosphate-activated protein kinase/nuclear factor-erythroid 2-related factor 2 signaling pathway / 中华皮肤科杂志

Objective:To evaluate the effect of metformin on ultraviolet A (UVA) -induced photoaging of an immortalized human keratinocytes cell line (HaCaT), and to explore its potential mechanisms.Methods:Cell counting kit 8 (CCK8) assay was performed to evaluate the effect of metformin at different concentrations (0 - 100 mmol/L) on the viability of HaCaT cells, and 10 mmol/L metformin was selected for subsequent experiments. Cultured HaCaT cells were divided into a blank control group (conventional culture), a metformin group (treated with culture medium containing 10 mmol/L metformin), a UVA irradiation group (conventional culture for 24 hours followed by 10 J/cm 2 UVA irradiation) and a metformin + UVA group (treated with culture medium containing 10 mmol/L metformin for 24 hours followed by 10 J/cm 2 UVA irradiation) ; UVA irradiation was performed at a dose of 10 J/cm 2 once a day for 3 consecutive days. After 4-day treatment, cells were collected, the β-galactosidase assay was performed to determine the proportion of senescent cells in each group, 2′, 7′-dichlorodihydrofluorescein diacetate assay to detect levels of intracellular reactive oxygen species (ROS), and the comet assay to detect DNA damage levels. Additionally, some HaCaT cells were divided into the blank control group, metformin group, 1.25 μmol/L dorsomorphin (an adenosine monophosphate-activated protein kinase [AMPK] inhibitor) + metformin group, and 2.5 μmol/L dorsomorphin + metformin group, and cells in the latter two groups were treated with 1.25 and 2.5 μmol/L dorsomorphin respectively for 2 hours, followed by the treatment with 10 mmol/L metformin for 24 hours. Western blot analysis was performed to determine the cellular localization and phosphorylation levels of nuclear factor-erythroid 2-related factor 2 (Nrf2). By using the small-interfering RNA (siRNA) -mediated silencing method, siRNA-Nrf2 was transfected into HaCaT cells to knock down Nrf2 expression (siRNA-Nrf2 group) ; 2.5 μmol/L dorsomorphin-treated HaCaT cells or Nrf2-knockdown HaCaT cells were treated with metformin and UVA irradiation (dorsomorphin + metformin + UVA group, siRNA-Nrf2 + metformin + UVA group, respectively), and the proportions of senescent cells were further calculated in each group. Statistical analysis was carried out by using one-way analysis of variance and two-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:Treatment with different concentrations of metformin for 24 hours could affect the viability of HaCaT cells to varying degrees ( F = 5 206.31, P < 0.001) ; there were no significant differences in the relative survival rates of HaCaT cells between the 10 - 20 mmol/L metformin groups and the control group (0 mmol/L metformin group, all P > 0.05), while the relative cell survival rates were significantly lower in the 25 - 100 mmol/L metformin groups than in the control group (all P < 0.05). After UVA irradiation, HaCaT cells shrank significantly and became narrow and elongated, and the intercellular spaces increased; the relative cell survival rate was significantly lower in the UVA irradiation group (76.13% ± 1.03%) than in the blank control group (100.00% ± 1.24%, LSD- t = 14.86, P < 0.001), but significantly higher in the metformin + UVA group (106.69% ± 2.45%) than in the UVA irradiation group (LSD- t = 11.55, P < 0.001). Moreover, the UVA irradiation group showed significantly increased proportions of senescent cells (45.14% ± 4.98%), intracellular ROS levels (144.61% ± 4.91%), and percentages of DNA in the tail (75.33% ± 1.77%) compared with the blank control group (23.84% ± 1.89%, 55.49% ± 1.57%, 1.88% ± 0.29%, respectively, all P < 0.001), while the metformin + UVA group showed significantly decreased proportions of senescent cells (24.26% ± 1.34%), intracellular ROS levels (58.62% ± 2.17%), percentages of DNA in the tail (15.83% ± 1.23%) compared with the UVA irradiation group (all P < 0.001). Western blot analysis showed that the Nrf2 expression in the cytoplasm was lower in the 10 mmol/L metformin group than in the blank control group, while the phosphorylated Nrf2 expression in the nuclei was higher in the 10 mmol/L metformin group than in the blank control group, suggesting that metformin could effectively induce the phosphorylation of Nrf2 and its nuclear translocation; both the pretreatment with 1.25 and 2.5 μmol/L dorsomorphin could significantly reduce the phosphorylation levels of AMPKα and Nrf2 induced by 10 mmol/L metformin. The proportions of senescent cells in the dorsomorphin + metformin + UVA group and the siRNA-Nrf2 + metformin + UVA group were 67.84% ± 2.74% and 65.94% ± 1.33%, respectively, which were significantly higher than those in the metformin + UVA group (37.76% ± 1.64%, t = 14.45, 13.34, respectively, both P < 0.001) . Conclusion:Metformin may inhibit UVA-induced photoaging of HaCaT cells by activating the AMPK/Nrf2 signaling pathway, scavenging ROS and reducing DNA damage.

Research progress of protease inhibitor Cystatin SN / 中国医师杂志

Cysteine protease inhibitor SN (CST1) is one of the members of type 2 Cystatin superfamily. It is widely expressed and distributed in mammals. It contains many types of CST proteins and is located on chromosome 20p11.2. Cystatin SN has a variety of biological functions and is involved in the occurrence and development of tumor genesis and metastasis, inflammation, cell cycle and aging.

Regulatory role of circIGF2BP3 in autophagy in photoaged dermal fibroblasts / 中华皮肤科杂志

Objective:To preliminarily investigate the effect of circIGF2BP3 on autophagy in photoaged dermal fibroblasts.Methods:Human dermal fibroblasts were isolated from circumcised foreskin tissues from 6 children in the Department of Urological Surgery, Third Affiliated Hospital of Sun Yat-sen University. An ultraviolet A (UVA) -induced photoaged human dermal fibroblast model (UVA radiation group) was established by repeated UVA radiation at a dose of 10 J/cm 2 for 14 consecutive days, and human dermal fibroblasts receiving no treatment served as control group. The photoaged cell model was verified by β-galactosidase staining, Western blot analysis for determining P21 protein expression, and cell counting kit-8 (CCK8) assay for evaluating cell viability. Moreover, Western blot analysis was performed to determine the protein expression of autophagy-related proteins P62, microtubule-associated protein 1 light chain 3 (LC3) -Ⅰand LC3-Ⅱ in photoaged human dermal fibroblasts, and real-time quantitative RCR (qRT-PCR) to verify the differential expression of circIGF2BP3 between photoaged and normal human dermal fibroblasts. Furthermore, circIGF2BP3 was biologically annotated. Some cultured primary human dermal fibroblasts were divided into 4 groups: empty vector group transfected with an empty vector, UVA + empty vector group transfected with an empty vector followed by repeated UVA radiation, circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector, UVA + circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector followed by repeated UVA radiation. Western blot analysis was performed to determine the expression of autophagy-related proteins. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference- t test. Results:Compared with the control group, the UVA radiation group showed significantly increased proportions of β-galactosidase-positive cells (61.33% ± 5.78% vs. 6.37% ± 0.32%, t = 9.49, P < 0.01) and P21 expression (1.25 ± 0.03 vs. 1.00 ± 0.05, t = 4.26, P < 0.05), but significantly decreased cell viability (74.33% ± 3.48% vs. 100%, t = 7.38, P < 0.01). Moreover, the P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly higher in the UVA radiation group than in the control group (both P < 0.05). The relative expression of circIGF2BP3 was 0.72 ± 0.04 in the photoaged human dermal fibroblasts, which was significantly lower than that in the normal human dermal fibroblasts (1.00 ± 0.03, t = 5.46, P < 0.01). The P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly lower in the circIGF2BP3 group (0.60 ± 0.01, 0.71 ± 0.01, respectively) than in the empty vector group (1.00 ± 0.02, 1.00 ± 0.01, t = 16.25, 2.75, P < 0.01, < 0.05, respectively), and lower in the UVA + circIGF2BP3 group (1.05 ± 0.02, 2.04 ± 0.05, respectively) than in the UVA + empty vector group (1.31 ± 0.02, 2.72 ± 0.14, t = 10.493, 6.472, respectively, both P < 0.01) . Conclusion:circIGF2BP3 can regulate autophagy in UVA-induced photoaged dermal fibroblasts, which provides a new potential therapeutic target for the prevention and treatment of skin photoaging.

Avaliação da Senescência de Células Sanguíneas Mononucleares Periféricas e na Disfunção Endotelial entre Adultos com Alto Risco Cardiovascular / Assessment of Peripheral Blood Mononuclear Cells Senescence and Endothelial Dysfunction among Adults with High Cardiovascular Risk

Resumo Fundamento Doenças cardiovasculares (DCV) são uma das principais causas de mortalidade e morbidade em todo o mundo. O envelhecimento biológico tem sido associado à ocorrência de resultados cardiovasculares. Entretanto, o mecanismo subjacente desse processo ainda é desconhecido. Objetivos Buscamos avaliar se a senescência das células sanguíneas mononucleares periféricas (CSMP) e biomarcadores endoteliais poderiam influenciar o risco cardiovascular (CV) e ser marcadores adequados para a detecção precoce de doenças cardiovasculares em adultos. Métodos Neste estudo transversal, pacientes livres de DCV foram classificados como baixo (n=32) e alto (n=28) escore de risco intracardaco (IHR) A senescência das CSMP foi avaliada estimando-se a atividade de telomerase (AT) e detectando-se a presença de células senescentes e disfunção endotelial, estimando-se a concentração de nitrito e nitrato e a capacidade antioxidante total (CAT). A análise estatística foi realizada com o software SPSS, versão 16.0 (SPSS Inc., Chicago, IL). Todos os p-valores <0,05 foram considerados estatisticamente significativos. Resultados A senescência de CSMP de 0,95 [p-valor = 0,0001; 95% IC (0,874-1,026)] foi um indicador significativo de pacientes com escore de IHR mais alto, com um valor de corte de 21,65, com sensibilidade e especificidade de 92% e 88% respectivamente. Identificou-se que a senescência de CSMP, nitrito e nitrato, e AT eram independentemente associadas a um escore de IHR alto. Conclusão Os status de nitrito e nitrato e AT, e a senescência de CSMP são medidas adequadas para prever o alto risco cardiovascular em adultos com risco CV. Entretanto devem ser realizados estudos de acompanhamento de longo prazo para confirmar esses achados. (Arq Bras Cardiol. 2021; 116(1):37-47)

Abstract Background Cardiovascular diseases (CVD) are one of the leading causes of mortality and morbidity worldwide. Biological aging has been associated with the occurrence of adverse cardiovascular outcomes; however, the underlying mechanism of this process remains unknown. Objectives This study sought to evaluate if peripheral blood mononuclear cell (PBMC) senescence and endothelial biomarkers could influence cardiovascular (CV) risk and be suitable markers for the early detection of cardiovascular diseases in adults. Methods In this cross-sectional study patients free of CVD were classified as lower (n=32) and higher Interheart Risk (IHR) scores (n=28). PBMC senescence was assessed by estimating the telomerase activity (TA) and detecting the presence of senescent cells and endothelial dysfunction by estimating the concentration of nitrite and nitrate and of total antioxidant capacity (TAC). Statistical analysis was performed with SPSS version 16.0 (SPSS Inc., Chicago, IL). All p-values <0.05 were considered statistically significant. Results PBMC senescence 0.95 [p-value = 0.0001; 95% CI (0.874-1.026)] was a significant predictor of patients with higher IHR scores with a cut-off value of 21.65 with a sensitivity and specificity of 92% and 88% respectively. PBMC senescence, nitrite and nitrate and TA were found to be independently associated with high IHR scores. Conclusion PBMC senescence, TA and nitrite, and nitrate status are suitable measures to predict high cardiovascular risk in adults with CV risk. Nevertheless, long-term follow-up studies are needed to confirm these findings. (Arq Bras Cardiol. 2021; 116(1):37-47)

Construction of an aging model of renal tubular epithelial cells / 中华老年医学杂志

Objective:To compare aging models for renal tubular epithelial cells induced by different drugs.Methods:Different concentrations of D-galactose(D-gal), hydrogen peroxide(H 2O 2)and cisplatin(CDDP)were administered to the human proximal tubular epithelial cell line(HK2). Cell activity and the half maximal inhibitory concentration(IC50)were measured by the CCK-8 assay; cell senescence was assessed by senescence-related β-galactosidase staining(SA-β-gal); senescence-related gene expression was detected by Western blotting; cell cycle distribution and apoptosis were determined by flow cytometry.Pathological changes in renal tubules and interstitial tissues were examined in D-gal-induced and naturally aging mice using HE staining, and p16 expression was detected using immunohistochemistry. Results:CCK-8 assay results showed that HK2 cell activity was inhibited treatment with each of the three compounds.The 48-hour IC50 values were(365.8±9.7)mmol/L for D-gal, (385.4±20.8)μmol/L for H 2O 2 and(8.4±1.6)μmol/L for CDDP.Light microscopic observation revealed slowed growth of HK2 cells in the three groups.The rate of SA-β-gal-positive cells increased, compared with the control group( P<0.05). Treatment resulted in an increase in G0/G1 phase cells by(22.9±1.0)% in the 400 mmol/L D-gal group and by(13.0±4.4)% in the 400 μmol/L H 2O 2 group, while G2/M phase cells increased by(14.4±1.9)%( t=48.07, 6.40, 16.53, P<0.05)in the 8 μmol/L CDDP group, compared with the control group.Also, compared with the control group, HK2 cell apoptosis increased by(50.3±1.0)% in the 400 μmol/L H 2O 2 group and by(41.9±2.0)% in the 8 μmol/L CDDP group, which was significantly higher than(7.7±0.4)% in the 400 mmol/L D-gal group( t=77.47, 33.73, 28.35, all P<0.05). Western blotting results indicated that the expression of CCND1 was down-regulated after any of the three drugs reached a certain concentration.The expression of p16 in the D-gal group was up-regulated( F=92.88, P<0.05), but there was no statistical difference in the expression of p16 after H 2O 2 or CDDP treatment.Mice of the D-gal model showed a decline in renal tubular cells, thickened basement membrane, widened interstitial spaces and increased expression of p16 in renal tubules similar to those observed in naturally aging mice. Conclusions:For HK2 cell senescence models induced by three different drugs, the renal tubular epithelial cell senescence model induced by D-gal is relatively close to the natural senescence model.

Role and action mechanism of microRNA-26a targeting EZH2 in ultraviolet A-induced photoaging of human skin fibroblasts / 中华皮肤科杂志

Objective:To investigate the expression of microRNA (miR) -26a in human skin fibroblasts during photoaging induced by ultraviolet A (UVA) , and to evaluate the effect of up-or down-regulation of miR-26a expression on the methylation level of the whole genome, the target gene enhancer of zeste homolog 2 (EZH2) and cell aging.Methods:Some human skin fibroblasts were irradiated with 10 J/cm 2 UVA once a day for 7 consecutive days, RNA was extracted on days 0, 3 and 7, and real-time quantitative reverse PCR (RT-PCR) was performed to determine the expression of miR-26a; miR-26a mimics and inhibitors were transfected into fibroblasts to up-or down-regulate the expression of miR-26a respectively, and fluorescence microscopy and RT-PCR were performed to determine the expression of miR-26a and evaluate the transfection efficiency. Some human skin fibroblasts were divided into 6 groups: blank control group receiving no treatment, UVA group treated with UVA irradiation according to the above method, miR-26a mimic group transfected with miR-26a-mimics, UVA+miR-26a mimic group transfected with miR-26a-mimics followed by UVA irradiation, miR-26a inhibitor group transfected with miR-26a inhibitors, UVA+miR-26a inhibitor group transfected with miR-26a inhibitors followed by UVA irradiation. On day 7, cells in each group were collected after the end of UVA irradiation. Then, flow cytometry was performed to detect cell cycle, DNA methylation quantitative detection kit was used to detect the methylation level of whole genome, RT-PCR was conducted to determine the mRNA expression of EZH2 (a histone-lysine N-methyltransferase enzyme) , DNA methyltransferase 1 (DNMT1) and miR-26a, and Western blot analysis was performed to determine the protein expression of EZH2 and DNMT1. Statistical analysis was carried out by using one-way analysis of variance and least significant difference- t test. Results:Compared with the unirradiated control group, the expression of miR-26a gradually increased in the UVA irradiation group over time during the culture, and there was a significant difference in the expression of miR-26a between the two groups after 7 days of UVA irradiation ( t=5.295, P < 0.05) . Strong fluorescence signals were observed in the miR-26a mimic-or miR-26a inhibitor-transfected fibroblasts, suggesting a high transfection efficiency. Flow cytometry showed that the proportion of cells at G1 phase significantly differed among the blank control group, UVA group, miR-26a mimic group, UVA+miR-26a mimic group, miR-26a inhibitor group, and UVA+miR-26a inhibitor group (52.82% ± 2.56%, 78.56% ± 4.34%, 53.63% ± 3.13%, 89.52% ± 4.17%, 54.39% ± 3.86%, 65.34% ± 4.78%, respectively; F=46.728, P < 0.01) , and significantly higher in the UVA group than in the blank control group ( t=8.848, P < 0.01) , higher in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group ( t=11.922, 3.154, P < 0.01, < 0.05, respectively) , and higher in the UVA+miR-26a inhibitor group than in the miR-26a-inhibitor group ( t=3.087, P < 0.05) , but significantly lower in the UVA+miR-26a inhibitor group than in the UVA group ( t=3.547, P < 0.05) . Detection of the genome-wide methylation level showed that the methylation level ( A450 value) significantly differed among the above groups (0.676 ± 0.024, 0.323 ± 0.043, 0.506 ± 0.035, 0.169 ± 0.024, 0.602 ± 0.036, 0.422 ± 0.029, respectively, F=97.402, P < 0.01) , and significantly lower in the UVA group than in the blank control group ( P < 0.01) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.01) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.01) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . RT-PCR and Western blot analysis showed significant differences in the mRNA and protein expression of EZH2 and DNMT1 respectively among the 6 groups (both P < 0.05) , which were significantly lower in the UVA group than in the blank control group ( P < 0.05) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.05) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.05) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . Conclusion:In the UVA irradiation-induced photoaging of skin fibroblasts, miR-26a expression was up-regulated, cellular proliferative activity and genome-wide methylation level decreased; up-regulation of miR-26a expression could down-regulate the expression of its target gene EZH2 and methylation-related gene DNM1, and promote cell photoaging, while down-regulation of miR-26a expression could up-regulate the expression of EZH2 and DNMT1, and inhibit cell photoaging.

Research progress of trabecular meshwork cell oxidative stress in the pathogenesis of glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

@#<p>Glaucoma is a common irreversible blinding eye disease, pathological elevated intraocular pressure is the main clinical feature. The formation of intraocular pressure, related to aqueous circulation, will be pathologically elevated when the aqueous cycle is abnormal. Trabecular network, which plays a key role in maintaining normal intraocular pressure, is the main component of aqueous outflow channel. Imbalance of oxidative stress manifested as oxidation and antioxidant effects is a direct risk factor for elevated intraocular pressure in glaucoma. When it comes to the trabecular meshwork cells, a series of changes such as deposition and degeneration of extracellular matrix, autophagy and aging will eventually occur, and finally the dysfunction of trabecular meshwork cells and increased aqueous outflow resistance, causing intraocular pressure pathological elevated. In this paper, we reviewed the research progress on the relationship between oxidative stress in trabecular meshwork cells and the pathogenesis of glaucoma, in order to provide evidence for further research and reference for exploring the pathogenesis, prevention and treatment of glaucoma.

Effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis. <p>METHODS: Forty cases(80 eyes)of grade II-Ⅳ CCH with liver-kidney Yin deficiency were randomly divided into two groups: combined treatment group and artificial tears group, which were treated with Qi Jing Mingmu decoction combined with artificial tears and simple artificial tears respectively. The international ocular surface disease index(OSDI), tear break-up time(BUT)and Schirmer Ⅰ test(SⅠt)were observed and the clinical effects were compared after 3mo treatment. For CCH patients with grade III or above, followed up for 3mo or more and willing to operate, the loose conjunctival tissue was removed and the cell aging related β-gal staining was performed on frozen sections. The results were statistically analyzed. <p>RESULTS: The OSDI score(14.53±2.68), BUT 9.25±3.02s and SⅠt(8.95±3.57mm/5min)of combined treatment group were significantly better than those of artificial tears group after 3mo treatment(all <i>P</i><0.05). After drug treatment, 7 cases(7 eyes)in artificial tears group and 4 cases(4 eyes)in combined treatment group of CCH patients were treated by operation. The positive rate of aging cells in combined treatment group was significantly lower than that in artificial tears group(16.00±7.84 <i>vs </i>39.00±14.09, <i>P</i>=0.013).<p>CONCLUSION: Qi Jing Mingmu decoction combined with artificial tears to treat CCH is more effective than simple artificial tears in relieving ocular symptoms, improving tear film and promoting tear secretion. Combined treatment can also reduce the cell aging in CCH, which can be used as a safe and effective treatment method in addition to surgical operation.

Effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis. <p>METHODS: Forty cases(80 eyes)of grade II-Ⅳ CCH with liver-kidney Yin deficiency were randomly divided into two groups: combined treatment group and artificial tears group, which were treated with Qi Jing Mingmu decoction combined with artificial tears and simple artificial tears respectively. The international ocular surface disease index(OSDI), tear break-up time(BUT)and Schirmer Ⅰ test(SⅠt)were observed and the clinical effects were compared after 3mo treatment. For CCH patients with grade III or above, followed up for 3mo or more and willing to operate, the loose conjunctival tissue was removed and the cell aging related β-gal staining was performed on frozen sections. The results were statistically analyzed. <p>RESULTS: The OSDI score(14.53±2.68), BUT 9.25±3.02s and SⅠt(8.95±3.57mm/5min)of combined treatment group were significantly better than those of artificial tears group after 3mo treatment(all <i>P</i><0.05). After drug treatment, 7 cases(7 eyes)in artificial tears group and 4 cases(4 eyes)in combined treatment group of CCH patients were treated by operation. The positive rate of aging cells in combined treatment group was significantly lower than that in artificial tears group(16.00±7.84 <i>vs </i>39.00±14.09, <i>P</i>=0.013).<p>CONCLUSION: Qi Jing Mingmu decoction combined with artificial tears to treat CCH is more effective than simple artificial tears in relieving ocular symptoms, improving tear film and promoting tear secretion. Combined treatment can also reduce the cell aging in CCH, which can be used as a safe and effective treatment method in addition to surgical operation.

Progress in cellular senescence and organ fibrosis / 中华老年医学杂志

Fibrosis as a pathogenic mechanism occurs in numerous organs and diseases.Persistent and severe fibrosis affect the function of the organ.Organ fibrosis is an important cause of patient morbidity and mortality.Aging is considered as a strong risk factor for development of organ fibrosis.Organ fibrosis is age-related disease and cellular senescence plays an important role in the occurrence and development of fibrosis,which provides a new strategy for the treatment of fibrosis.This review is focused on the progress in cellular senescence and organ fibrosis.

Cellular senescence-mediated skin aging / 中华皮肤科杂志

Aging is age-related degeneration of the whole body,and skin aging is one of the most visualized changes in aging.Cellular senescence is a stress response to stable cell cycle arrest,and it is both the hallmark of aging and the important mechanism of the occurrence and development of aging.With the development of skin aging,senescent cells gradually accumulate in both the epidermis and dennis,and further exacerbate aging.Thus,when these senescent cells are eliminated,the aging skin seems to be rejuvenated.Cellular senescence is involved in the process of skin aging,which may be related to activation of DNA damage response pathway,up-regulation of regulatory proteins blocking cell cycle,and increase of senescence-associated secretory phenotypes.Cellular senescence is expected to be a novel target for preventing skin aging in the future.

A study on alterations in mitochondrial biological characteristics during cellular senescence of human embryonic lung fibroblasts / 中华预防医学杂志

Objective@#To study the alterations of mitochondrial biological characteristics during both cellular replicative and premature senescence induced by hydrogen peroxide in human embryonic lung fibroblasts (HEFs).@*Methods@#The premature senescence was induced by 400 μmol/L H2O2 once a day at the same time and with 2 hours each time, after four consecutive days the premature senescence models were classified into premature senescence initiation group (PSi) and premature senescence persistence group (PSp). Based on the life span of HEFs, the cell replicative senescence was divided into five groups included young-age (22 PDL), middle-age (35 PDL), replicative senescence (49 PDL), PSi and PSp. The mitochondrial distribution, relative content, adenosine triphosphate (ATP) contents, 8-hydroxydeoxyguanosine (8-OHdG) levels, the relative mitochondrial transcription factor A (TFAM) as well as mitochondrial DNA methyltransferase 1 (mtDNMT1) mRNA levels, mtDNA copy number, the relative TFAM protein level and the total enzyme activity of mitochondrial DNA methyltransferases (mtDNMTs) were detected in five senescence groups.@*Results@#The mtDNA copy number, 8-OHdG contents, level of mtDNMT1 mRNA and mtDNMTs activity in 49 PDL group were higher than those in 22 PDL group (all P values <0.05); The level of 8-OHdG in PSi was higher than that in 22 PDL group (P<0.05); The ATP contents, mtDNA copy number, the mRNA and protein expression levels of TFAM and mtDNMTs activity of PSp were higher than those in 22 PDL group (all P values<0.05).@*Conclusion@#During the cellular senescence of HEFs, the higher mtDNA copy number and mtDNMTs activity were common features regardless of replicative or premature senescence, with possibility that oxidative stress was involved in modifying the occurrence of premature senescence.

Antagonistic effect of platelet-rich plasma on photo-aged skin in rat / 中华医学美学美容杂志

Objective To observe the effect of platelet-rich plasma (PRP) on photoaging skin of rat.Methods F344 rats were used as model animals and skin photoaging model was established by UV irradiation.The model animals were divided into 5 groups:group A was injected with activated PRP (A-PRP);group B was injected with inactive PRP (N-PRP);group C was injected with normal saline;group D was irradiated only with UV;Four weeks after the injection,the appearance and histological characteristics of the rat's skin were evaluated.Results Compared with group C and group D,the wrinkles and skin color in group A and group B were significantly improved.Histological observation showed that the histological features of group A and group B were significantly better than that of group C and D (P＜0.01).The thickness of dermis in group A was higher than that in group B (P＜0.05).There was no significant difference between groups C and D (P＞0.05).The thickness of dermis in group E was significantly higher than that in group C (P＜0.01).Conclusions Injection of PRP can improve the appearance and histological features of photo-aged skin in rats.A-PRP is better than N-PRP.

Effect of photoaging on the degradation of advanced glycation end products by human dermal fibroblasts / 中华皮肤科杂志

Objective To evaluate the effect of photoaging on the degradation of advanced glycation end products (AGEs) by human dermal fibroblasts.Methods Some cultured human dermal fibroblasts were subjected to repetitive ultraviolet A (UVA) radiation (UVA radiation group) to establish a photoaging cell model,which was then evaluated by cell counting kit 8 (CCK-8) assay,senescenceassociated β-galactosidase staining and detection of apoptosis rate.Moreover,fibroblasts receiving no treatment served as control group.Some other primary fibroblasts were divided into 4 groups:photoaged group receiving UVA radiation,non-photoaged group receiving no treatment,AGE-treated photoaged group treated with UVA radiation followed by the treatment with 200 mg/L AGE-bovine serum albumin (BSA),and AGE-treated non-photoaged group treated with 200 mg/L AGE-BSA alone.After the treatment with AGE-BSA for 4-72 hours,flow cytometry was performed to determine the fluorescence intensity of AGE-BSA in fibroblasts of the above groups.After 8-hour treatment with AGE-BSA,confocal laser scanning microscopy was performed to localize and semiquantitatively detect AGE-BSA in fibroblasts,and enzymelinked immunosorbent assay (ELISA) was conducted to detect AGE-BSA levels in fibroblasts,as well as changes in the intracellular AGE-BSA level within 24 hours after the removal of AGE-BSA.Results Compared with the control group,the UVA radiation group showed significantly decreased cellular proliferative activity (t =7.559,P ＜ 0.05),but significantly increased apoptosis rate and percentage of β-galactosidase-positive fibroblasts (t =14.075,43.524 respectively,both P ＜ 0.05).Flow cytometry revealed that the average fluorescence intensities of AGE-BSA after 4-,8-,16-,24-,48-and 72-hour treatment with AGE-BSA were significantly higher in the AGE-treated photoaged group (293.00 ± 8.19,359.67 ± 11.59,347.00 ± 12.29,338.00 ± 12.77,334.67 ± 14.22 and 336.30 ± 10.21,respectively) than in the photoaged group (all P ＜ 0.05),as well as in the AGE-treated non-photoaged group (222.33 ± 8.74,276.33 ± 6.11,256.33 ± 5.51,243.00 ± 10.15,236.33 ± 1.53 and 240.33 ± 1.52,respectively) than in the non-photoaged group (all P ＜ 0.05).Moreover,the average fluorescence intensities of AGE-BSA at different time points were all significantly higher in the AGE-treated photoaged group than in the AGE-treated non-photoaged group (all P ＜ 0.05).Confocal laser scanning microscopy showed that AGE-BSA was mainly localized in lysosomes after endocytic uptake into the fibroblasts,and the AGE-treated photoaged group showed significantly increased fluorescence intensity of AGE-BSA compared with the AGE-treated non-photoaged group (P ＜ 0.05).ELISA revealed that the intracellular AGE level in the AGE-treated non-photoaged group at 24 hours after the removal of AGE-BSA was decreased by (14.6 ± 1.2)％ compared with that before the removal,and the degradation rate of AGE-BSA was significantly higher in the AGE-treated non-photoaged group than in the AGE-treated photoaged group (7.6％ ± 1.4％,t =6.604,P ＜ 0.05).Conclusion The internalized AGE-degradating ability decreases in photoaged fibroblasts,which may induce the accumulation of AGEs in photoaged skin.

Jinfukang induced senescence of circulating tumor cells in lung cancer by p16/RB signal pathway / 肿瘤

Objective: To explore the molecular mechanism of Jinfukang inducing the senescence of circulating tumor cells (CTCs) in patients with non-small cell lung cancer. Methods: Human lung cancer CTCs (CTC-TJH-01 cells) were treated with different concentrations (0, 125, 250, 500 and 1 000 μg/mL) of Jinfukang. The proliferation of CTC-TJH-01 cells was detected by CCK-8 assay. The state of cellular senescence was detected by β-galactosidase staining. The expression of proliferating cell nuclear antigen (PCNA) protein in CTC-TJH-01 cells was detected by flow cytometry and immunofluorescence staining. The expression levels of proliferation- and senescence-associated proteins including p16, p21, retinoblastoma protein (RB) and phospho-RB (p-RB) were detected by Western blotting. Results: Jinfukang inhibited the proliferation of CTC-TJH-01 cells (500 and 1 000 μg/mL: both P < 0.05), induced the senescence of CTC-TJH-01 cells (350 and 700 μg/mL: both P < 0.05), and promoted the expression of PCNA protein in CTC-TJH-01 cells (350 and 700 μg/mL: both P < 0.05). The expression levels of p16 and p21 proteins were up-regulated, but the expression level of p-RB protein was down-regulated after treatment (all P < 0.05). Conclusion: Jinfukang induces the senescence of CTCs by regulating p16/RB signal pathway, which may be the mechanism of preventing the recurrence and metastasis of lung cancer.

Mechanism of cellular senescence in acute kidney injury that leads to chronic kidney disease in elderly mice / 中华肾脏病杂志

Objective To investigate the mechanism of cellular senescence in ischemia reperfusion injury (IRI)-induced acute kidney injury (AKI) that leads to chronic kidney disease (CKD) in elderly mice.Methods An acute kidney injury model was established in C57B1/6 male mice at ages 8-10 weeks (young group) or 20-24 months (old group) by bilateral IRI.The animals were randomly divided into 4 groups as follows:Young-Sham (n=8),Old-Sham (n=8),Young-IRI (n=8),and Old-IRI groups (n=8).All mice were weighted,and their blood was collected from the tail vein at days 1,3,and 7 after surgery.The mice were killed on day 14 after surgery,and their kidneys were harvested for further analysis.Serum was used for the creatinine test.The changes of the renal tissue morphology and pathology were assessed using hematoxylin-eosin staining and sirius red staining.Immunofluorescence staining of collagen Ⅰ,F4/80,phosphor-histone H3 (p-HH3),and Ki67 were performed to determine the stage of the collagen deposit,macrophage filtration,and cell cycle G2/M arrest.The collagen Ⅰ expression was analyzed using western blot.The expression levels of TNF-α,IL-6,TGF-β,and collagen Ⅰ were determined using real-time PCR.Results Compared with that in the sham group,the serum creatinine levels in both Young-IRI and Old-IR1 groups were obviously increased.The Young-IRI group recovered completely on day 7.The Old-IRI group had higher creatinine levels than the Young-IRI group at each time point.Morphology and pathology analyses revealed that acute injury was repaired in the Young-IRI group,but slight inflammatory cell filtration and collagen deposition were observed in the Old-Sham and Old-IRI groups,respectively.Immunofluorescence staining revealed some F4/80-positive macrophage filtration,collagen Ⅰ deposition,and p-HH3 and Ki67 double-positive nuclear tubular epithelial cells in the Old-Sham group,but considerably more positive results were found in the Old-IRI group.Western blot analysis revealed that collagen Ⅰ expression level was higher in the Old-IRI group than in the Young-IRI group (P ＜ 0.01) and in the Old-Sham group than in the Young-Sham group (P ＜ 0.05).Real-time PCR demonstrated that the mRNA expression levels of cytokines and fibrosis markers,including of TNF-α,IL-6,TGF-β,and collagen Ⅰ,in the Old-Sham and Old-IRI groups were increased as compared with those in the Young-Sham and Young-IRI groups (P ＜ 0.05).Conclusions The levels of kidney inflammation,fibrosis,and cell-cycle arrest are lower in the old mice.After IRI injury,a sustained and ongoing inflammatory reaction is involved and more cells are arrested in the cell cycle G2/M,which inhibit renal repair and promote fibrosis progression.