Objective To observe expression of Caspase-3 and apoptosis around the prosthesis and explore the relationship of the expression and the apoptosis with the periimplant osteolysis. Methods From April 2001 to August 2006, 16 patients (10 males, 6 females) underwent the revision total hip arthroplasty surgery, who had the primary total hip arthroplasty at the ages of 45-67 years and had the revision total hip arthroplasty at the ages of 55-78 years, with the implantation duration of 7-13 years. According to their preoperative X-ray films andthe findings during the operation, the patients were divided into two groups: theloose/osteolytic group (n=8) and the loose/non-osteolytic group (n=8). The interface tissues were obtained from the peri-implant region in the patients. The synovial samples were taken from another 6 patients (2 males, 4 females; age, 54-68years; illness course, 9-15 years), who underwent the primary total hip arthroplasty for osteoarthritis. These 6 patients were used as controls. The tissues were prepared for the immunohistochemical assays to determine the expression of Caspase-3. The TUNEL assays were performed to quantify the apoptotic cells. The quantitative analysis on the positive cells and the correlation with the presence of the particulate wear debris and the severity of osteolysis were also performed. Results The level of the expression for Caspase-3 and the apoptosis index inthe loose/osteolytic group were significantly increased when compared with those in the loose/non-osteolytic group and the control group (P＜0.01). The polyethylene particles were surrounded by more positive cells than the metal particles. The positive cells were present at a higher level in the tissue sections where the high-wear status was present when compared with the areas where the low-wear status was present (P＜0.05). Conclusion There is a statistical correlation of the Caspase-3 expression to the apoptosis index and to the presence of the particulate wear debris and the severity of osteolysis, which may be one of the key points for the bone reconstruction inhibition and the bone resorption at the boneimplant interface under the stimulation of the wear debris. The apoptosis is involved in the pathogenesis of the aseptic loosening, which is closely related to the signal transportation of Caspase-3.
Objective To evaluate the phenomena of apoptosis and its relevant mechanism during ischemia-reperfusion period. Methods The published papers to explore the apoptotic phenomena and its mechanism in organs or tissues which experienced ischemia-reperfusion injury were reviewed. Results Apoptosis was common in ischemia-reperfusioned organ or tissue. The severity of apoptosis was influenced by many factors such as ischemia, hypoxia, oxygen free radials, intracellular free calcium ion overloading, various cytokines, et al; and also was regulated by bcl-2 family, caspase family and NF-κB，et al. Conclusion Apoptosis is a common phenomenum in ischemiareperfusioned organ or tissue which is affected and regulated by various factors.
【Abstract】Objective To investigate the apoptosis induced by TGF-β1 in human hepatic carcinoma cell lines and its relationship with p53 gene and Smad. Methods Three human hepatic carcinoma cell lines which involving in various status of the p53 gene were used in this study. TGF-β1-induced apoptosis in hepatic carcinoma cell lines was measured by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. To study the mechanism of TGF-β1-induced apoptosis, these cell lines were transfected with a TGF-β1-inducible luciferase reporter plasmid containing Smad 4 binding elements (SBE) and luciferase gene using Lipofectamine 2000, then treated with TGF-β1, relative luciferase activity was assayed. Results Of three cell lines studied with TUNEL assay, TGF-β1 induced apoptosis was observed in HepG2 cells (wild type p53). Huh-7 (mutant p53) and Hep3B (deleted p53) cell lines showed less apoptosis. Luciferase activity assay indicated that the response to TGF-β1 induction in HepG2 cells was increased dramatically but was not significant in Huh-7 and Hep3B cell lines. Conclusion HepG2 cells seem to be highly susceptible to TGF-β1-induced apoptosis compared with Hep3B and Huh7 cell lines. Smad 4 is a central mediator of the TGF-β1 signal transduction pathway.
OBJECTIVE:To investigate the cytotoxic effects of homoharringtonine(HHT) on HXO-RB44 cell line and the cell death form induced byHHT in vitro. METHODS:MTT assay was adopted to establish survival rate of the tumor cells. Agarose gel electrophores was chosen to detect the genomic DNA from the cells exposed to HHT. RESULTS:In the concentration from 10-9 to10-4 mol/L HHT powerfully inhibited the growth of the cells (P＜0.05). Regular genomic DNA fragmentation from the cells exposed to 10-6mol/L HHT for 48 hours was shown to be typical DNA ladder on agarose gelelectrophoresis. CONCLUSION :HHT can induce retinoblastoma (RB) programmed cell death (PCD),the effects of which has close correlation with incubated period and concentration of HHT.
Objective To study the effect and mechanism of the apoptosis of hypertrophic scar fibroblasts (HSF) induced by artesunate(Art). Methods HSFs were isolated and cultured from human earlobe scars by the tissue adherence method. The 3th to 5th generation cells were harvested and divided into two groups. HSF was cultured with normal medium in control group and with medium containing60, 120 and 240 mg/L (5 ml)Art in experimental group. Apoptosis and cell cycle were identified by light microscopy, electronmicroscopy and flow cytometry. Then, HSF was cultured with normal medium in control group and with medium containing 30, 60 and 120 mg/L Art in experimental group. The changes of intracellular calcium concentration were observed. Results The primary HSF was fusiform in shape and adherent. The vimentin positive expression was analyzed by immunocytochemistry. Art could induce apoptosis of HSF in the range of 60-240 mg/L under inverted microscope. The effect was dose and timedependent. Clumping of nuclear chromatin showed margination in the experimentalgroup. And the disaggregation of the nucleolus were observed under electronmicroscopy. There were significant differences in the proportion of HSF apoptosis and HSF at G0-G1,S, G2-M stages between the two groups(P＜0.05). Apoptotic peak was shown in experimental group by flow cytometry. The peak became more evident asArt concentration increased. The intracellular calcium concentration elevated markedly in HSF with 30-120 mg/L Art treatment for 24 hours, showing significant differences between the two groups (P＜0.05). Conclusion The Art facilitates HSF cells apoptosis in vitro by the change of cell cycle. It is suggested that intracellular calcium variation may be one of the mechanisms of HSF apoptosis induced by Art.
Objective To analyze the hotspots and development trends in the research field of tumor cell apoptosis and autophagy. Methods Relevant literature on tumor apoptosis and autophagy published between January 2012 and December 2021 were searched through the Web of Science core collection database, and CiteSpace 5.8.R3 software and VOSviewer version 1.6.10 software were used to analyze the country/region, institution, keywords and citation node information of the literature. Results A total of 6716 foreign-language articles were included in the study after searching and screening, and the number of papers showed a linear upward trend year by year. China published the largest number of articles and cooperated closely with other research institutions, but there were not many high-quality and influential articles. The two journals Autophagy and Cell were more authoritative in the field of tumor apoptosis and autophagy research. The signaling pathways and related proteins of apoptosis and autophagy, and the study of tumor suppressor mechanisms based on apoptosis/autophagy were current research hot topics. The migration, apoptosis and epithelial mesenchymal transformation of cancer cells would be the research focus and direction in the future. Conclusions In the past 10 years, the research on tumor apoptosis and autophagy has continued to develop. With the in-depth research on the molecular level, the study of its mechanism is expected to further reveal the mystery of tumor apoptosis and autophagy.
PURPOSES:To investigate the time of neuronie apoptosis in the retinas of Imman fetuses,and its relations with neuronie proliferation and differentiation, METHODS:The retinas of 27 human fetuses from 8th to 38th week of R,~til- ization age and 3 adults were studied by TdT-mediated dUTP nick end labelling(TUNEL) method. RESULTS:Tbe nuctei of labeled apoptotic cells were charaeterised by nuclear marginization,ehromatln condensation and cleseent shape,and some apoptotie bodies were visible in the specimens. The apoptosis of neuroepithelium of fetal rclina took place during 8th to 18th week, Apoptosis of ganglion cells were observed from 1256 to 18th week. The apoptos[s of pholorec, plors were formd from 14th to 2Ist week ,while thai of bipolar neurones and M~ller cells were found from ldth to 28th week. No apoptosb of ocstones were observed in the retinas after 28th week of fertilization age and within the retinas of adults. CONCLUSION:The proliferating cells of neuroepithelium and Ihe neurones which just differetiated from fetal retina might partly undergo apoptosis. The time of apoptosls of differentiated neurones was consistent with the time of the synapses formation between neurones and their targel cells. （Chin J Ocul Fundus Dis,1997,13:67 -69 ）
Objective To observe the biological characters of chondrocytes in articular loose body and to find out seeding cells for cartilage tissue engineering. Methods Samples from 5 loose body cartilages, 2 normal articular cartilages and 6 osteoarthritis articular cartilages were collected. Part of each sample’s cartilage was histologically studied to observe the chondrocytes distribution the morphologic changes by toluidine-blue staining, chondrocytes’ apoptosis by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL). The rest of each cartilage was digested and isolated by 0.25% trypsin and 0.2% collagenase Ⅱ, and then were cultivated in 10%DMEM. Their morphologic changes were observed 24h later.Comparison was made btween three cartilages. Results Compared with normal cartilage and osteoarthritis articular cartilage, the cells density was higher, their lacunars were larger, cells distribution was irregular, and apoptosis was more apparent in loose body cartilage. Conclusion The characters of chondrocytes from loose body is more like fibroblasts so they can not serve as seeding cells directly for cartilage tissue engineering.
Objective To investigate the protective effects and the mechanism of recombinant human growth hormone on the intestinal barrier function. Methods The literatures of recent years were reviewed and summarized． Results The recombinant human growth hormone not only prevent mucosal cells and immunological cells from apoptosis, but also antagonize the damage of NO, cytokines, as well as endotoxin on intestinal barrier. What’s more, it increases the intestinal uptake and utilization of glutamine. All of the above could maintain the integrity and functions of the intestinal barrier. Conclusion The recombinant human growth hormone protects the intestinal barrier function through different ways.
Objective To investigate the effects of recombinant human erythropoietin ( rHuEPO) on expressions of Bax and Bcl-2 proteins in hyperoxia-induced lung injury of adult rats. Methods Fortyeight healthy male SD adult rats were randomly divided into six groups. The control group ( 0 h) breathed with room air. The rHuEPO intervention group was put into oxygen chamber and breathed with 100% O2 for 96 h plus intraperitoneal injection of rHuEPO (1000 U/kg) daily. Other four groups were put into oxygen chamber and breathed with 100% O2 for 24, 48, 72 and 96 h respectively. Arterial blood gases were measured to calculate oxygenation index. Wet-to-dry weight ratios of left lung were measured. The contents of TNF-α and IL-1β in bronchoalveolar lavage fluid (BALF) were assayed with radioimmunoassay. The expressions of Bax and Bcl-2 proteins in the lung were determined withWestern blot and immunohistochemisty. The changes of lung histopathology were assessed by hematoxylin and eosin stain and observed under light microscope. Results After breathing 100% O2 , the oxygenation index decreased gradually and reached minimal value at 96 h. The wet-to-dry weight ratio of left lung increased gradually and reached maximal value at 96 h. The contents of TNF-α and IL-1β in BALF reached maximal value at 48 h and then decreased gradually. The expression of Bax protein increased, but the expression of Bcl-2 protein decreased gradually in the lung. Compared with the 96 h group, the oxygenation index was higher, wet-to-dry weight ratio and contents of TNF-α and IL-1β in BALF decreased, the expression of Bax protein decreased, and the expression of Bcl-2 protein increased in the lung of the rHuEPO group. Conclusion rHuEPO can attenuate hyperoxia-induced lung injury of adult rats by down-regulating expression of Bax protein and up-regulating expression of Bcl-2 protein.