Application of cell derivative in periodontal regeneration / 中国组织工程研究

BACKGROUND:Cell derivative is cell-derived bioactive components,including decellularized extracellular matrix,exosome,apoptotic extracellular vesicle,and conditioned medium,has the effects on immune regulation,promoting angiogenesis,bone regeneration,ligament remodeling,and is capable of promoting stem cell chemotherapy,migration,proliferation,and adhesion.Its excellent characteristics make it a promising biomaterial for application and clinical translation in the field of periodontal tissue engineering. OBJECTIVE:To review the characteristics of cell derivatives(decellularized extracellular matrix,exosome,apoptotic extracellular vesicle,and conditioned medium)and its effect and the latest progress in the field of regenerative restoration of periodontal complex tissue structures. METHODS:We searched the articles on CNKI and PubMed databases with the search terms"regeneration,periodontal tissue,tissue engineering,decellularized matrix,exosome,apoptotic extracellular vesicle,condition medium"in Chinese and English,respectively.Finally,76 articles were included for analysis and discussion. RESULTS AND CONCLUSION:(1)Among those four cell derivatives,the decellularized extracellular matrix has the best mechanical properties and fibrous structure,serving as a biomimetic scaffold to provide physiochemical signals and participate in mechanical signaling in periodontal tissue engineering,providing supporting effect suitable for periodontal regeneration.Recently,the development of soluble decellularized extracellular matrix bioinks has enabled the fabrication of regenerative scaffolds for personalized periodontal defects.(2)Exosomes are the simplest cell derivatives that have immunomodulatory capacity,promoting cell migration and differentiation.As a carrier,they can be used to carry target molecules to regulate periodontal regeneration,promote ligament remodeling and bone regeneration,and are suitable for periodontal tissue engineering.(3)Apoptotic vesicles generated from apoptotic cells have a strong immunomodulatory effect and can recruit stem cells and macrophages,which determine the fate of stem cells through signal transduction and can enhance immunomodulation to promote periodontal regeneration.Engineered extracellular vesicle is considered to have the potential to initiate targeted internal immunomodulation.(4)The extraction of conditioned medium is simple and completely noninvasive,which provides essential nutrients and growth factors for tissue regeneration.These components are crucial for successful periodontal regeneration.Therefore,the conditioned medium is especially suitable for studying the interactions between cells in vitro and has an important role in high-throughput detection in the future.

Effects of electroacupuncture combined with transcranial alternating current stimulation on inflammatory-apoptotic genes expression of MCAO rats / 中国康复医学杂志

Objective:To establish right middle cerebral artery occlusion(MCAO)model in rats and to investigate the mechanism underlying motor function regulation by transcranial alternating current stimulation(tACS)intervention.Furthermore,to dynamically observe the effects of electroacupuncture combined with tACS on neurological defi-cit scores(NDS),cerebral blood flow,inflammatory-cell apoptosis gene of MCAO model rats and to explore the mechanism of cerebral and neural regulation on motor function rehabilitation after cerebral ischemia reperfusion. Method:Forty SD rats were randomly divided into sham-operation group(S group),model group(M group),electroacupuncture group(EA group),transcranial alternating current stimulation group(T group)and electroacupuncture combined with transcranial alternating current stimulation group(EA+T group).After 2 h ischemia-reperfusion,EA group was given bilateral Qu chi(LI 11)and Zu san li(ST 36)electroacupuncture under anesthesia.Right Ml was selected for tACS in T group.EA+T group was treated with EA and tACS to-gether.S and M group were treated with anesthesia for 30min per time,for 7 days.The data from before modling(B)to after modling(D7)were recorded,including neurological deficit score and blood flow of the right middle cerebral artery by laser doppler flowmetry.RT-PCR was used to analyze inflammation-cell apopto-sis gene expression at D7. Result:Neurological deficit score:at 2h,D1,M group,EA group,T group and EA+T group increased signifi-cantly compared with other time(P＜0.05).At D3,D5,D7,S group decreased significantly compared with oth-er time(P＜0.05),while M group increased significantly compared with other group(P＜0.05).EA group,T group and EA+T group were significantly different in all times(P＜0.05).At 2h,D1,D3,D5,D7,M group increased significantly compared with B.At D1,D3,D5,D7,NDS decreased significantly compared with that at 2h(P＜0.05).Blood flow:EA+T group and S group decreased at 2h,increased at D1 and decreased at D3.EA group increased at D3.However,M group decreased significantly compared with other group at D1,D3 and D5(P＜0.05).RT-PCR:motor cortex △Ct analysis:Caspase 12 in EA+T group decreased significantly compared with T group(P＜0.05),IL-1β and NLRPla in EA group and T group decreased significantly com-pared with those in S group(P＜0.05).2-△△Ct analysis in ischemic region:ATF4 in M group increased significant-ly compared with that in other groups(P＜0.05);Bcl 2 in M group increased significantly compared with that in S group,EA group and EA+T group(P＜0.05);Bax,Caspase 12,C-fos in M group increased significantly compared with those in S Group(P＜0.05). Conclusion:Electroacupuncture combined with tACS can modulate the inflammatory response and inhibit cell apoptosis through regulating ATF4、Bcl-2、Bax、Caspase 12、C-fos and can be a new strategy for the treatment of ischemic stroke.

Research progress of BCL-2 family apoptotic regulation and its mediated drug resistance after antitumor drug therapy / 中国胸心血管外科临床杂志

@#Apoptosis is an important means to regulate cell proliferation and maintain homeostasis. Recent researches have shown that the B-cell lymphoma-2 (BCL-2) family not only plays a dominant role in the regulation of normal cell apoptosis, but also plays a crucial role in the formation of tumor genesis, progression and subsequent drug resistance mediated by the escape mode of apoptosis. The phenomenon that BCL-2 family antagonized the apoptosis induced by antitumor drugs and then acquired drug resistance has been reported in the clinical treatment of hematologic lymphatic system tumors, breast cancer, lung cancer, gastric cancer and other diseases. Thus, specific inhibitors targeting anti-apoptotic members of the BCL-2 family have emerged with the development of research. In this paper, we systematically reviewed the regulation of apoptosis mediated by BCL-2 family and the drug resistance mediated by BCL-2 family. Meanwhile, we summarized the research advances of BCL-2 family specific inhibitors to provide new strategy for solving the problems on tumor therapeutic resistance and for finding new therapeutic targets in the future.

Cytotoxic and Apoptotic Properties of the Flavonoid-rich Ethyl Acetate Fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells

Introduction@#Breast cancer is the most common cancer among women in the Philippines and about 3 in every 100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer. @*Objectives@#This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines. @*Methods@#Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining. @*Results@#The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses. @*Conclusion@#The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.

Effect of cadmium chloride toxicity on olfactory mucosa / Efecto de la toxicidad del cloruro de cadmio en la mucosa olfativa

SUMMARY: Cadmium is a highly toxic metal and affects the respiratory mucosa. The aim of the study is to show the inflammation and degenerative effect of cadmium on the olfactory mucosa. In this study, eight-week-old Wistar rats with an average weight of 170-190 g were divided into two groups (control and experiment) with 20 animals in each group and used in the experiments. The rats in the experimental group were given 2 mg/kg/day powdered cadmium chloride dissolved in water intraperitoneally every day for two weeks. At the end of the experiment, the nasal cavity was completely removed with anesthesia. Concha nasalis superior was separated, fixed with zinc-Formalin solution and decalcified with 5 % EDTA (Ethylene-diaminetetraacetic acid). After routine histopathological procedure, APAF-1 antibody was used for expression of Hematoxylin-Eosin (HE) and immunohistochemistry. Histopathological examination revealed interruptions in the basement membrane structure due to cadmium and degenerative changes in stem cells, degeneration in sensory cells and pycnosis in nuclei, dilatation in blood vessels and increased inflammation in connective tissue. APAF-1 expression was found to increase in epithelial cells and olfactory glands (Bowman gland) cells. It has been thought that cadmium toxicity increases cell degeneration and inflammation in the olfactory mucosa and may significantly affect cell death and olfactory metabolism by inducing the pro-apoptotic process.

El cadmio es un metal altamente tóxico que afecta la mucosa respiratoria. El objetivo fue mostrar el efecto inflamatorio y degenerativo del cadmio sobre la mucosa olfativa. En este estudio, ratas Wistar de ocho semanas de edad con un peso promedio de 170-190 g se dividieron en dos grupos (control y experimental) con 20 animales en cada grupo. Las ratas del grupo experimental recibieron 2 mg/kg/día de cloruro de cadmio en polvo disuelto en agua por vía intraperitoneal todos los días durante dos semanas. En los animales se exirpó la cavidad nasal bajo anestesia. Se separó la concha nasal superior, se fijó con solución de zinc-Formalina y se descalcificó con EDTA (ácido etilendiaminotetraacético) al 5 %. Después del procedimiento histopatológico de rutina, Hematoxilina- Eosina (HE) e inmunohistoquímica, se utilizó el anticuerpo APAF-1. El examen histopatológico reveló interrupciones en la estructura de la membrana basal debido al cadmio y cambios degenerativos en las células madre, degeneración en las células sensoriales y picnosis en los núcleos, dilatación de los vasos sanguíneos y aumento de la inflamación en el tejido conjuntivo. Se encontró que la expresión de APAF-1 aumenta en las células epiteliales y en las células de las glándulas olfatorias (glándulas de Bowman). Se ha pensado que la toxicidad del cadmio aumenta la degeneración celular y la inflamación en la mucosa olfativa y puede afectar significativamente la muerte celular y el metabolismo olfativo al inducir el proceso proapoptótico.

Vitamina C y su acción en células cancerígenas

RESUMEN Introducción: La vitamina C es una sustancia que desde hace ya varios años ha suscitado debate debido a la cantidad de usos que se han demostrado. En algunos casos, las utilidades han ido desde profilaxis y acortamiento de la duración de resfriados, hasta estudios de acción en enfermedades, tales como el cáncer; los mecanismos de acción de esta han sido evaluados en forma de monoterapia o en combinación con quimioterapia para demostrar o descartar su utilidad en el cáncer. Objetivo: Demostrar si los efectos de la vitamina C fueron efectivos y si su uso, solo o en combinación con quimioterapia, es de utilidad. Método: Se realizó una investigación de tipo descriptiva documental, realizada con artículos científicos en el periodo comprendido desde 2016 hasta 2021, con un análisis detallado de los resultados del uso de la vitamina C y su posible efecto sobre los diferentes tipos de cáncer. Fueron buscados en las bases de datos de SciELO, Scopus y Medline. Resultados: La información hallada fue organizada según concentraciones plasmáticas de vitamina C y su acción en células cancerosas, dosis evaluadas de la vitamina C, mecanismos de acción en relación a células cancerígenas, desequilibrio redox, efecto específico en cánceres, vitamina C y cáncer de mama. Conclusiones: En la revisión realizada se evidencia que la vitamina C tiene un efecto benéfico en los cánceres hematopoyéticos, como: leucemia, melanoma, cáncer de mama o ciertos tipos de cáncer colorrectal y que disminuyen los efectos adversos producidos por medicamentos quimioterapéuticos.

ABSTRACT Introduction: Vitamin C is a water-soluble substance that has been in debate since a long time ago due to its wide demonstrated use. In some cases, its usage have ranged from prophylaxis and shortening the duration of colds, to studies of its action in diseases, such as cancer; Its mechanisms of action have been evaluated in the form of monotherapy or in combination with the chemotherapy to demonstrate or rule out its usefulness in cancer. Objective: To demonstrate whether the effects of vitamin C were effective and whether its use, alone or in combination with chemotherapy, is useful. Method: A documentary descriptive research was carried out, supported with scientific articles (period 2016 to 2021) which analyze in detail the results of the use of vitamin C and its possible effect on different types of cancer. Results: The information found was organized according to plasma concentrations of vitamin C, its action on cancer cells, evaluated doses of vitamin C, mechanisms of action in relation to cancer cells, redox imbalance, specific effect on cancers, vitamin C and breast cancer. Conclusions: The review shows that the use of vitamin C has a beneficial effect on hematopoietic cancers, such as leukemia, melanoma, breast cancer or certain types of colorectal cancer, and also reduces the adverse effects produced by chemotherapeutic drugs.

RESUMO Introdução: A vitamina C é uma substância que há vários anos desperta o debate devido ao número de usos que foram demonstrados. Em alguns casos, os benefícios vão desde a profilaxia e redução da duração dos resfriados, até estudos de ação em doenças, como o câncer; os mecanismos de ação deste foram avaliados como monoterapia ou em combinação com quimioterapia para provar ou descartar sua utilidade no câncer. Objetivo: Demonstrar se os efeitos da vitamina C foram eficazes e se seu uso, isoladamente ou em combinação com a quimioterapia, é útil. Método: Foi realizada uma pesquisa documental descritiva, realizada com artigos científicos no período de 2016 a 2021, com análise detalhada dos resultados do uso da vitamina C e seu possível efeito nos diferentes tipos de câncer. Eles foram pesquisados nas bases de dados SciELO, Scopus e Medline. Resultados: As informações encontradas foram organizadas de acordo com as concentrações plasmáticas de vitamina C e sua ação nas células cancerígenas, doses avaliadas de vitamina C, mecanismos de ação em relação às células cancerígenas, desequilíbrio redox, efeito específico sobre cânceres, vitamina C e câncer de mama. Conclusões: Na revisão realizada, fica evidente que a vitamina C tem efeito benéfico sobre os cânceres hematopoiéticos, como: leucemia, melanoma, câncer de mama ou certos tipos de câncer colorretal e que reduz os efeitos adversos produzidos pelos quimioterápicos.

A cross-sectional and histological analysis to understand the cytological effects of cell phone radiation on buccal mucosa of children

Context: The ongoing pandemic has affected all the spheres of life and one of the severely affected avenues is the education of a child. The online education has seen an upward curve since the start of COVID-19 pandemic. Schools globally have adopted online class tutorials as the main method to impart education and directly increasing the screen time for a child. Aim: The aim of the present study was to evaluate the cytological effects of prolonged mobile phone usage on the buccal mucosa of children. Settings and Design: Stratified sampling was used for the selection of subjects for the study. After a questionnaire regarding the usage of a mobile phone was distributed among the parents of children. Among them, 90 children were selected on the basis of pattern and frequency of mobile phone usage in the child. Materials and Methodology: The children were divided into three groups based on the per day hours of viewing of mobile phone, i.e., Group 1: Usage of 1–2 h a day, Group 2: Usage of 3–6 h a day, and Group 3: Usage of >6 h a day. The time frame taken into consideration was 1 year after the pandemic started. This was specifically to understand the impact of the online education. Swab was obtained by using the conventional ice-cream stick method from the buccal mucosa. Statistical Analysis: The samples were subjected to histological and microscopical analysis to observe for cytological changes. One-way ANOVA was used to determine the statistical significance if any. Results: The results obtained clearly showed that Group 3 (>6 h usage per day) showed the highest number of cellular and chromosomal aberrations which was significant. Conclusion: The results indicated that impact due to the prolonged screen time on the buccal mucosa is significant. A direct proportionality was seen between the apoptotic changes and chromosomal aberrations and the number of daily hour usage.

Dental pulp stem cell-derived apoptotic bodies regulate macrophage polarization and inflammatory response / 口腔疾病防治

Objective@#To investigate the effects of apoptotic bodies (ABs) derived from dental pulp stem cells (DPSCs) on macrophage polarization and inflammation response in vivo. @*Methods @#Human DPSCs were extracted, cultured and identified. Staurosporine was used to apoptosis induction and differential methods were performed for ABs identification. The in vitro cultured macrophages were divided into 3 groups: solvent control, lipopolysaccharide (LPS), and the LPS+ABs. The macrophages were stimulated with LPS to induce inflammation followed by ABs treatment. In the untreated group, macrophages were added with an equal amount of solvent. The specific uptake of ABs by macrophages, the expression level of CD206 and the levels of inflammatory cytokines were analyzed. The mouse models of cutaneous wounds and dextran sulfate sodium (DSS)-induced colitis were established, and the mice were randomly divided into 3 groups: the PBS-treated group, the DPSCs-treated group, and the ABs-treated group. The mice were injected with the same volume of PBS, DPSCs and ABs, respectively. The body weight, histological pathology, the expression levels of CD206 and cytokines, and the extent of tissue regeneration were measured.@* Results @#DPSCs and ABs derived from DPSCs were successfully isolated and characterized. ABs could be taken up by macrophage. While lipopolysaccharide(LPS) induced production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), ABs significantly reduced the levels of these pro-inflammatory cytokines and increased the expression of transforming growth factor-β (TGF-β) and CD206 (P < 0.01). In the cutaneous inflammatory wound model, the wound closure rate in mice intravenously injected with ABs was significantly accelerated (P < 0.05). The administration of ABs markedly reduced the pro-inflammatory factors levels and increased the CD206+ cell number. In the colitis model, treatment with ABs markedly reduced the loss in bodyweight (P < 0.05), recovered the colon length (P < 0.01), and significantly increased the CD206+ cell number.@* Conclusion@# DPSCs-derived ABs could enhance macrophage M2 polarization and attenuate inflammation. Therefore, ABs could be used as a promising cell replacement for inflammatory regulation and tissue regeneration.

Oxidative Damage to BV2 Cells by Trichloroacetic Acid: Protective Role of Boron via the p53 Pathway / 生物医学与环境科学（英文）

This study aimed to investigate the neurotoxicity induced by trichloroacetic acid (TCA) and the possible protective mechanisms of boron (B). Mouse BV2 cells were treated with TCA (0, 0.39, 0.78, 1.56, 3.12, 6.25, or 12.5 mmol/L) and B (0, 7.8, 15.6, 31.25, 62.5, 125, 500, or 1,000 mmol/L) for 3 h and 24 h, respectively. Then, reactive oxygen species, and supernatant proinflammatory cytokine and protein levels were analyzed after 24 h of combined exposure. Beyond the dose-dependent decrease in the cellular viability, it clearly increased after B supplementation ( P < 0.05). Moreover, B decreased oxidative damage, and significantly down-regulated IL-6 levels and up-regulated TNF-β production ( P < 0.05). B also decreased apoptosis via the p53 pathway. The present findings indicated that TCA may induce oxidative damage, whereas B mitigates these adverse effects by decreasing cell apoptosis.

Effect of Hedysarum Polysaccharides on Bcl-2/Caspase-3 Signaling Pathway of Schwann Cells Cultured in High Glucose / 中国实验方剂学杂志

ObjectiveTo observe the effects of Hedysarum polysaccharides（HPS）on the signaling pathways of B-cell lymphoma 2 （Bcl-2）， cysteinyl aspartate-specific protease 3 （Caspase-3）， and Bcl-2-associated X protein （Bax） in Schwann cells（SCs）cultured in high glucose，and explore the possible mechanism of HPS against diabetic peripheral neuropathy（DPN）. MethodFour SD suckling mice aged 5-7 days were randomly divided into a normal group，a high-glucose group，an HPS + high-glucose group，and an α-lipoic acid（α-LA）+ high-glucose group. SCs were extracted from the sciatic nerve and cultured in a 37 ℃，5% CO2 incubator. After the cells reached 80% confluence，Cell Counting Kit-8（CCK-8）was used to screen the experimental concentrations suitable for high glucose，HPS， and α-LA interventions. Western blot and Real-time polymerase chain reaction （Real-time PCR）were used to detect the protein and mRNA expression of Bcl-2，Bax，and Caspase-3. The apoptosis rate of SCs was detected by flow cytometry using Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI）. ResultAs revealed by Western blot and real-time PCR，compared with the normal group，the high-glucose group showed reduced protein and mRNA expression of Bcl-2 and increased protein and mRNA expression of Bax and Caspase-3（P<0.01）. Compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed increased protein and mRNA expression of Bcl-2 and decreased protein and mRNA expression of Bax and Caspase-3（P<0.01）. As displayed by the results of flow cytometry using Annexin V/PI， compared with the normal group，the high-glucose group showed increased apoptosis rate；compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed reduced apoptosis rate（P<0.01）. ConclusionHPS can alleviate the apoptotic response of SCs，and its mechanism may be related to the inhibition of the activation of the Bcl-2/Caspase-3 signaling pathway.

Evaluation of antitrypanosomal properties and apoptotic effects of ochrolifuanine from Dyera costulata (Miq.) Hook.f against Trypanosoma brucei brucei

@#Trypanosoma brucei parasites are flagellated kinetoplastid protozoan which is responsible for Human African Trypanosomiasis (HAT). Current chemotherapy drugs have a number of side effects and drug resistance has emerged as a major issue in current treatment. Active bisindole alkaloid compound ochrolifuanine was previously isolated from the leaves of Dyera costulata. In vitro antitrypanosomal activity of ochrolifuanine against Trypanosoma brucei brucei strain BS221 showed strong activity with an IC50 value of 0.05 ± 0.01 µg/ml. We compared the effect of ochrolifuanine and reference compound staurosporine in T. b. brucei apoptosis. The apoptosis-inducing activity of ochrolifuanine was evaluated using TUNEL assay and cell cycle analysis. Trypanosoma brucei brucei was shown to undergo apoptotic cells death as demonstrated by the appearance of several conical hallmarks of apoptosis. Ochrolifuanine was found to induce apoptosis in parasites in a dose- and time-dependent manner. The cell cycle study revealed 0.025 and 0.05 µg/ml of ochrolifuanine arrested the growth of T. b. brucei at two different growth phases (G0/G1 and in S phases). While at concentration 0.10 µg/ml arrested at the G2/M phase. In conclusion, the results indicate that ochrolifuanine displayed an antitrypanosomal effect on T. b. brucei by inducing apoptosis cell death and causing the arrest of parasite cells at different growth phases. The results suggested that ochrolifuanine may be a promising lead compound for the development of new chemotherapies for African trypanosomiasis.

The mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of acute pancreatitis cells / 中华急诊医学杂志

Objective:To explore the mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of pancreatic cells and participating in the occurrence and development of acute pancreatitis.Methods:Pancreatic acinar cells AR42J from rats were treated by caerulein, and then the levels of amylase, tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1) and IL-6 in the supernatant of cell culture were detected by ELISA to verify the cell model of acute pancreatitis. RT-PCR was used to detect the expression of miR-494 in normal AR42J cells (control group) and acute pancreatitis cell model (model group). Flow cytometry was used to detect the apoptosis of the control group, negative control miRNA transfected acute pancreatitis cell model (negative control group) and miR-494 transfected acute pancreatitis cell model (miR-494 transfection group). Western blot was used to detect the expression of ROCK1 and PTEN in the control group, negative control group and miR-494 transfection group.Results:The levels of amylase, TNF-α, IL-1 and IL-6 in the supernatant of AR42J cells treated with caerulein for 8 h and 12 h were significantly higher than those at 0 h and the control group ( P<0.05), indicating that the model was successfully constructed. The expression levels of miR-494 at 8 h, 12 h and 24 h after the establishment of acute pancreatitis cell model were significantly higher than those at 4 h and the control group ( P < 0.05). The apoptosis rate of the model group was significantly higher than that of the control group ( P<0.05), and the apoptosis rate of the miR-494 transfection group was significantly lower than that of the model group ( P<0.05). The expression levels of ROCK1 and PTEN in the miR-494 transfection group were significantly lower than those in the model group and negative control group ( P<0.05). Conclusions:When acute pancreatitis occurs, overexpression of miR-494 can inhibit the expression of pro-apoptotic protein, thus inhibiting the apoptosis of pancreatic acinar cells and promoting the development of acute pancreatitis.

All-trans retinoic acid-induced apoptosis signaling pathway in ARPE-19 cells / 中华实验眼科杂志

Objective:To investigate all-trans retinoic acid(ATRA)-induced apoptosis signaling pathway in ARPE-19 cells in vitro. Methods:The APRE-19 cell was treated with different concentrations of ATRA for 24 hours and 48 hours.Cell counting kit-8 (CCK-8) was used to detect the cell viability in order to determine the experimental concentration range.Flow cytometry and Western blot method were performed to evaluate the apoptosis and caspase related protein levels in ARPE-19 cells treated with 0, 2.5, 5, 10, 15 and 20 μmol/L of ATRA for 24 hours.Flow cytometry was used to detect the reactive oxygen species (ROS) and multicaspase levels and quantitative real-time PCR was carried out to determine the mRNA relative expression levels of caspase related proteins in ARPE-19 cells treated with 0, 2.5, 5, 10 and 20 μmol/L of ATRA, and 0 μmol/L ATRA group was used as the blank control group.Results:CCK-8 test showed that the half maximal inhibitory concentration of ARPE-19 cells treated with different concentrations of ATRA for 24 hours and 48 hours were 13.88 μmol/L and 11.99 μmol/L, respectively.The cell survival rates of ARPE-19 cells treated with different concentrations of ATRA for 24 hours and 48 hours were significantly different ( F=176.60, 350.30; both at P<0.01). When cultured for 24 hours, the cell survival rates of ARPE-19 cells in the 2 μmol/L and 6 μmol/L of ATRA groups were higher than that of the blank control group (both at P<0.05), and the cell survival rates of ARPE-19 cells in the 12, 14, 16, 18 and 20 μmol/L of ATRA groups were lower than that of the blank control group (all at P<0.05). Flow cytometry showed that there were significant differences in the apoptosis, ROS and multicaspase level among ARPE-19 cell groups treated with different concentrations of ATRA ( F=86.39, 116.84, 101.40; all at P<0.01). The apoptosis rates of APRE-19 cells in the 2.5 μmol/L and 5 μmol/L of ATRA groups were significantly decreased than that of the blank control group, and the apoptosis rate of APRE-19 cells in the 10, 15 and 20 μmol/L of ATRA groups were significantly increaseded than that of the blank control group (all at P<0.01). The relative expression levels of multicaspase and ROS were significantly higher in the 2.5, 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). Western blot assay showed that the relative expression level of caspase 9 was increased in the 2.5, 5, 10, 15 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.05). Compared with the blank control group, the relative expression levels of caspase 12 were increased in the 2.5 μmol/L of ATRA group and reduced gradually in the 5, 10, 15 and 20 μmol/L of ATRA groups, among which there were significant differences between the blank control group and 2.5, 15, and 20 μmol/L of ATRA groups (all at P<0.05). The relative expression level of caspase 3 was significantly increased in the 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.05). The relative expression level of cleaved caspase 3 was significantly increased in the 2.5, 5, 10, 15 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). Quantitative real-time PCR assay showed that the relative expression levels of caspase 9 and caspase 12 mRNA were significantly higher in the 2.5, 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). The relative expression levels of caspase 3 mRNA were significantly higher in the 5 μmol/L and 10 μmol/L of ATRA groups than that of the blank control group (both at P<0.01). When the concentration of ATRA was lower than 10 μmol/L, the relative expression levels of caspase 9 and caspase 12 mRNA were elevated in a concentration-dependent manner.When the concentration of ATRA reached 20 μmol/L, the relative expression levels of caspase 9 and caspase 12 mRNA were markedly decreased, but it was still higher than that of the blank control group. Conclusions:ATRA induces apoptosis in ARPE-19 cells in vitro through activating the reactive oxygen species and endogenous caspase-dependent apoptotic pathway.

Network pharmacology analysis and experimental validation to explore mechanism of Shenlian extract on myocardial ischemia / 中国药理学与毒理学杂志

OBJECTIVE To explore the potential molecular mechanism of Shenlian (SL) on myocardial ischemia (MI) on the basis of network pharmacology. METHODS Firstly, the main active ingredients of SL were screened in the Traditional Chinese Medicine Integrated Database, and the MI-associated targets were collected from the DisGeNET database. Then, we used compound-target and target-pathway networks to uncover the therapeutic mechanisms of SL. On the basis of network pharmacology analysis results, we assessed the effects of SL in MI rat model and oxygen glu?cose deprivation model of H9c2 cells and validated the possible molecular mechanisms of SL on myocardial injury in vivo and in vitro. RESULTS The network pharmacology results showed that 37 potential targets were recognized, including TNF-α, Bcl-2, STAT3, PI3K, and MMP2. The pathways revealed that the possible targets of SL were involved in the reg?ulation of inflammation and apoptosis signaling pathway. Then, in vivo experiments indicated that SL significantly reduced the myocardial infarction size of MI rats. Serum CK-MB, cTnT, CK, LDH, and AST levels were significantly decreased by SL (P<0.05 or P<0.01). In vitro, SL significantly increased H9c2 cell viability. The levels of inflammation factors including TNF-α and MMP2 were significantly decreased by SL (P<0.05 or P<0.01). TUNEL and Annexin V/propidium iodide assays indicated that SL could significantly decrease the cell apoptotic rate in vivo and in vitro (P<0.05 or P<0.01). The remarkable upregulation of anti-apoptotic Bcl-2 and downregulation of pro-apoptotic Bax protein level further confirmed this result. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the PI3K-Akt and JAK2-STAT3 pathways were significantly enriched in SL. Compared with the model group, SL treatment significantly activated the PI3K-Akt and JAK2-STAT3 pathways in vivo and in vitro according to Western blotting analyses. CONCLU?SION SL could protect the myocardium from MI injury. The underlying mechanism may be related to the reduction of inflammation and apoptosis by activating the PI3K/Akt and JAK2/STAT3 pathways.

Effect of calycosin in reducing cerebral ischemia/reperfusion injury in rats by regulating cytochrome C/apoptotic protease activating factor-1 apoptosis signal pathway / 解剖学报

Objective To investigate the effect of calycosin on cerebral ischemia/reperfusion injury and its mechanism. Methods Forty SPF male SD rats were randomly divided into sham group, model group, calycosin group (20 mg/kg), nimodipine group (0.7 mg/kg, positive control group). The occlusion model of middle cerebral artery in rats was established by modified thread occlusion method, and the environment of cerebral ischemia-reperfusion injury was simulated in vivo. Zea longa score was used to detect the neurological deficit of rats after ischemia-reperfusion injury, 2, 3, 5-triphenyltetranitrogen (TTC) was used to detect the volume of cerebral infarction, HE staining was used to detect the pathomorphological changes of nerve cells, Nissl staining was used to observe the changes of nissl bodies, TUNEL staining was used to detect the apoptosis of nerve cells, Western blotting was used to detect the expression of cytochrome C (Cyt C), apoptotic protease activating factor-1 (Apaf-1), Caspase-9 and Caspase-3. Results Compared with the sham group, the neurological deficit symptoms in the model group were significant (P<0.05), the volume of cerebral infarction increased significantly (P<0.05). Under the microscope, it was found that the nerve cells showed contraction of cell body, hyperchromatic and pyknosis of nucleus and poor growth state, the expression of nissl body reduced significantly (P < 0.05), the apoptotic nerve increased significantly (P< 0.05), the expression of Cyt C, Apaf-1, Caspase-9 and Caspase-3 increased significantly (P<0.05). Compared with the model group, the neurological deficit symptoms of calycosin group and nimodipine group reduced significantly (P<0.05), the volume of cerebral infarction reduced significantly (P<0.05). Under the microscope, the damage of nerve cells reduced significantly, the expression of nissl body increased significantly (P<0.05), the apoptotic nerve reduced significantly (P<0.05), the expression of Cyt C, Apaf-1, Caspase-9 and Caspase-3 decreased significantly (P<0.05). Conclusion Calycosin can significantly inhibit the apoptosis of nerve cells and reduce the cerebral ischemia-reperfusion injury. Its mechanism of action is related to the effective regulation of Cyt C/Apaf-1 apoptosis signaling pathway by calycosin.

Effect of osteoclast-derived apoptotic body microRNA-30a on osteogenic activity / 解剖学报

Objective To explore that whether apoptotic bodies released by osteoclasts mediate osteogenic activity. Methods The osteoclasts were induced from mouse (n = 10) bone marrow monocytes in vitro, and were identified by tartrate resistant acid phosphatase (TRAP) staining, F-actin, and DAPI double labeling immunofluorescence. The Co- culture system of osteoclasts and mouse osteoblasts MC-3T3E1 was established. The apoptosis of osteoclasts was analyzed by DNA fragment ELISA. Immunoblotting of apoptotic body markers was investigated. Real-time PCR analysis of bone formation markers was tested. MiRNA expression profiling of apoptotic body was identisfied. Results Alendronate (ALN) 100 μmol/L induced osteoclast apoptosis and caused apoptotic body release from osteoclasts. The expression of C3b and annexin V protein was enhanced by ALN; the expression of C3b in osteoclasts was negatively correlated with the activity of osteoblasts; the microarray screening of apoptotic body showed that miR-30a was correlated with bone formation markers and serum alkaline phosphatase (ALP). Conclusion Osteoclast-derived apoptotic body miR-30a can inhibit the activity of osteoblasts. Apoptotic body may participate in the dialogue between osteoclasts and osteoblasts.

Clinicopathological study of premalignant and malignant lesions of cervix along with apoptotic index and Ki-67 expression

Background: Cervical cancer is known to have a good response to radiotherapy. The response and prognosis are dependent on the level of apoptosis. Pap smear and histopathology are cost-effective methods in diagnosing premalignant and malignant lesions of cervix but not accurate in classifying and estimating the progression of the disease, especially in premalignant lesions. Therefore this study was undertaken to know the role of Ki-67 expression and apoptotic index in classifying accurately the premalignant lesions for better management.Methods: The study included 540 cases diagnosed histologically as cervical intraepithelial neoplasia or carcinoma. The apoptotic index is calculated for all the 540 cases using light microscopy on Haematoxylin and Eosin stained sections. Ki-67 immunohistochemical staining was done for 100 cervical biopsies. Ki-67 expression was graded and the Ki-67 labelling index was calculated. Statistical evaluation was done using the unpaired t-test.Results: The Apoptotic index increased with increasing grade of dysplasia. There is a significant difference in the mean apoptotic index between premalignant and malignant lesions of the cervix. The ki-67 index increased with increasing grade of dysplasia. There is a significant difference in the mean Ki-67 index between premalignant and malignant lesions of the cervix.Conclusions: Apoptotic index and proliferative indices have been found useful in distinguishing between premalignant and malignant lesions of the cervix and gives an idea about the proliferative activity of the tumour for better management of the patient and to determine prognosis.

Esclerosis sistémica y neoplasias, a propósito de un estudio en la provincia de Holguín / Systemic Sclerosis and neoplastic purpose of a study at Holguin province

La esclerosis sistémica (ES) es una enfermedad de causa desconocida, que se caracteriza por una producción exagerada de moléculas que componen la matriz extracelular. La disminución en la producción de óxido nítrico por las células endoteliales de la microvasculatura parece desempeñar un papel central en la patogenia de la enfermedad. Los resultados alcanzados en un estudio de serie de casos de un universo de 44 pacientes y muestra de 31, con baja incidencia de las causas neoplásicas en la muerte y como reacciones secundarias a tratamiento inmunosupresor con ciclofosfamida, según la conducta terapéutica aplicada, fue el motivo para la presentación de este trabajo, que reflejó la posible relación entre la esclerosis sistémica y las neoplasias. Se concluyó que la relación entre autoinmunidad y cáncer puede ser el resultado de un origen etiológico común (genético, hormonal, metabólico o factores ambientales) o un mecanismo de síndrome paraneoplásico. La enfermedad es terreno de riesgo para la ocurrencia de neoplasias, así como las neoplasias pueden inducir ES(AU)

The systemic sclerosis (SS) is a disease of unknown cause, that the fact that they fix the extra-cell womb characterizes itself for a production exaggerated of molecules. The decrease in the production of nitric oxide for the microvasculature's endothelial cells seems to play a central role in the pathogenesis of the disease. The results attained in 44 patients' study of series of cases of universe and inmunosupresor with cyclophosphamide according to therapeutic applied conduct shows of 31, with low incidence of the causes neoplastic in the death like secondary reactions and to treatment you went from motivation for the presentation of this work, that you reflected the possible relation between the systemic sclerosis and the neoplastic. It was concluded that the relation between auto-immunity and cancer can stem from an etiological common origin (genetic, hormonal, metabolic or environmental factors) or a mechanism of syndrome paraneoplastic. The disease is earthly of risk for neoplastic funny remark, the same way that the neoplastic can induce SS(AU)

Menthol enhances interleukin-13-induced synthesis and secretion of mucin 5AC in human bronchial epithelial cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of interleukin (IL) -13 combined with cold stimulation on synthesis and secretion of mucin (MUC) 5AC in human bronchial epithelial cell line 16HBE and explore the role of transient receptor potential 8 (TRPM8) and anti-apoptotic factor B-cell lymphoblast-2 (Bcl-2) in this process.@*METHODS@#16HBE cells were stimulated with 10 ng/mL IL-13, 1 mmol/L menthol, or both (1 mmol/L menthol was added after 6 days of IL-13 stimulation), and the changes in the expression of MUC5AC, intracellular Ca@*RESULTS@#The mRNA and protein expressions of MUC5AC increased significantly in 16HBE cells following stimulation with IL-13, menthol, and both (@*CONCLUSIONS@#Menthol combined with IL-13 produces a synergistic effect to promote the synthesis and secretion of MUC5AC in 16HBE cells possibly by activating TRPM8 receptor to upregulate the expression of Bcl-2.

Effect of Yisui Jiedu Prescription on Apoptosis of Hippocampal Neurons Based onPI3K/Akt/Bad Signal Pathway in Rats with Vascular Dementia / 中国实验方剂学杂志

Objective:To investigate the effect of Yisui Jiedu prescription on hippocampal neuron damage in vascular dementia (VD) rats and to regulate phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/recombinant Bcl-2 associated death promoter (Bad) mechanisms of signaling pathways of neuronal apoptosis. Method:The 40 SD rats were divided into sham operation group， model group， donepezil hydrochloride group and Yisui Jiedu prescription group， with 10 rats in each group.VD animal model was prepared by bilateral carotid artery permanent ligation (2-VO) method.The sham operation group and the model group were intragastrically administered with normal saline， the donepezil hydrochloride group was intragastrically administered with donepezil hydrochloride 0.52 mg·kg-1. The Yisui Jiedu prescription group was administered with Yisui Jiedu prescription (11.11 g·kg-1)， 1 time/d . After 30 days， Morris water maze was used to test the learning and memory ability of rats， hematoxylin-eosin (HE) staining was used to observe the histomorphological structure of hippocampal CA1 region. Ultrasound of neuron in rat hippocampal CA1 region was observed by transmission electron microscopy (TEM).Real-time fluorescent quantitative（Real-time PCR） was used to detect the Akt， Bad mRNA expression.Western blot was used to detect the Akt， p-Akt and Bad protein expression in hippocampus. Result:Compared with sham operation group， the learning and memory ability of model group decreased significantly(P<0.05). The pathological structure and neuronal ultrastructure of the hippocampus were changed obviously. Hippocampal tissue Akt mRNA and the Akt，p-Akt protein expression level decreased significantly(P<0.05)， and the levels of Bad mRNA and protein were significantly increased (P<0.05). Compared with model group， Yisui Jiedu prescription group can significantly improve the learning and memory ability of rats， improve the neuronal cells and ultrastructural changes in hippocampal CA1 area，and increase the expression of Akt mRNA and Akt，p-Akt protein in hippocampus. Decreased Bad mRNA and Bad protein expression levels (P<0.05). Conclusion:Yisui Jiedu prescription can significantly improve the learning and memory ability of VD rats， improve the ultrastructural pathological changes of hippocampus and neurons， and repair damaged neurons， which may promote Akt phosphorylation and activate PI3K/Akt/Bad. The signaling pathway plays a role in the defense of neurons against apoptosis.