No abstract available.
Animals* ; Brain* ; Hypothermia* ; In Situ Nick-End Labeling* ; Models, Animal*

AIMTo investigate the relationship between germ cell degeneration and apoptosis in cryptorchid rats.

METHODSThirteen 21-day-old Wistar rats were made unilaterally cryptorchid by closing the left inguinal canal. At day 30 (Group 1, n=6) and day 60 (Group 2, n=7) after operation, the testes were removed for histopathological examination. The controls (n=8) were sham operated and were sacrificed at day 60. Germ cell apoptosis was assessed by means of the TUNEL method.

RESULTSSpermatogenesis was arrested and the testicular and seminiferous tubular diameters were significantly reduced In the unilateral undescended testes (UUTs) compared with the contralateral descended testes (CDTs) and the control rats. However, atrophic changes, pathological calcification, necrosis of seminiferous tubule, and absence or sloughing of germ cells were not found in all the animals. The spermatocytes were the main type of germ cells undergoing apoptosis in all the groups. In the UUTs, there was a significant and time-dependent increase in the mean apoptotic index. By 60 days after surgery, increased apoptosis in germ cells was also observed in the CDTs.

CONCLUSIONApoptosis is the predominant mechanism of germ cell death rather than atrophy and necrosis in cryptorchidism.

Animals ; Apoptosis ; Cryptorchidism ; pathology ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Wistar ; Spermatocytes ; pathology

OBJECTIVETo observe the change of quantitative distribution,apoptosis and proliferation of T and B cells in the skin of KM mutant mice.

METHODSWe chose 1-,3-,6-,9-,22-day,3-,6-month-old KM mutant and wild-type mice to detect the changes of T and B lymphocytes using blood routine tests and immunohistochemical staining. Apoptosis was detected by TUNEL staining and proliferation by proliferating cell nuclear antigen (PCNA) staining.

RESULTST cells on KM mutant mice skin were mainly seen in epidermis and dermis. They increased on the first day to 6(th) day after birth and decreased on the 9(th) and 22(nd) day,but after 3-month-old,their number began to increase;at the time of 6 months,the number of B cells also increased. The apoptosis of the skin hair follicle and sebaceous gland cells were more obvious in KM mutant mice than in wild-type mice,with the maximal apoptosis occurred at the age of 22-day-old in both groups. The proliferation of epidermal basal cells in KM mutant mice between 1 to 9-day-old was not significantly different from that in the wild-type mice,but decreasing on the 22(nd) day and 3(rd) month and increasing in the 6(th) month. The proliferation in hair follicle and sebaceous glands decreased on 9(th) day,increased on 22(nd) day,and deceased on the 3(rd) month again.

CONCLUSIONSThe quantitative distribution,apoptosis,and proliferation of T and B lymphocytes abnormally change in the skin tissue of KM spontaneous mutant mice. They may lead to immune and hair growth disorders and promote the inflammatory responses.

Animals ; Apoptosis ; B-Lymphocytes ; Cell Proliferation ; In Situ Nick-End Labeling ; Mice ; Skin ; T-Lymphocytes

OBJECTIVETo explore the influences of Mycobacterium tuberculosis on the levels of human acute monocytic leukemia cell line THP-1 apoptosis and death.

METHODSHuman acute monocytic leukemia cell line THP-1 were infected with Mycobacterium tuberculosis strains H37Ra, H37Rv, or Beijing genotype (BJTB), respectively, to construct the infection models. Cell apoptosis was detected using flow cytometry. The distribution of the apoptotic proteins was detected using immunofluorescent staining assays. The cells late apoptosis was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining assays. The change of cell death was determined by Tyrpan blue staining assays.

RESULTSTHP-1 apoptosis was induced by Mycobacterium tuberculosis strains H37Ra, H37Rv, and BJTB. H37Ra strongly induced THP-1 apoptosis, H37Rv weakly induced THP-1 apoptosis, and BJTB induced THP-1 apoptosis at the lowest level among these three Mycobacterium tuberculosis strains. On the contrary, BJTB strongly induced THP-1 death, H37Rv weakly induced THP-1 death, and H37Ra induced THP-1 death at the lowest level.

CONCLUSIONSMycobacterial strains with different virulence induce different levels of apoptosis and death of THP-1 cells. Compared with highly virulent strains, attenuated strains induce more apoptosis and less death.

Apoptosis ; Cell Line, Tumor ; Humans ; In Situ Nick-End Labeling ; Leukemia, Monocytic, Acute ; Mycobacterium tuberculosis ; pathogenicity ; Virulence

PURPOSE: This study was aimed to evaluate changes in the stromal keratocyte after ablation of 50 micrometer and 100 micrometer with use of photorefractive keratectomy(PRK). METHODS: At 4 hours, 24 hours, 72 hours, 7 days and 1 month after PRK, each group of rabbits including normal control group was treated with terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling(TUNEL) staining using ApopTag(R) kit in vivo, then apoptotic keratocytes were evaluated with light microscope. RESULTS: There was no response with TUNEL staining of the epithelial cells, stromal keratocyte, and endothelium in normal cornea. In the ablation group, however, regardless of the depth of photorefractive ablation, the TUNEL signal was maximal after 4 hours, and it decreased with time. The signal was more intense in 100 micrometer ablation group than 50 micrometer ablation group, although the signal was not observed at the endothelial cells in both groups. The number of apoptotic stromal keratocytes at each time point of 4 hr, 24 hr, 72 hr, and 1 week was 57+/-8.9, 49+/-7.5, 36+/-5.1, and 12+/-1.3 cells/field in 100 micrometer ablation group, and 31+/-4.4, 28+/-4.6, 21+/-3.9, and 5+/-1.1 cells/field in 50 micrometer ablation group, and the difference between the two groups was statistically significant(P<0.05). CONCLUSION: The more the amount of ablation with photorefractive keratectomy, the stronger the apoptotic response. The postoperative apoptotic response was observed especially within 1 week. These findings suggest that early suppression of postoperative apoptosis within 1 week will influence on the prognosis of visual quality after photorefractive keratectomy, and more studies will be needed in the future.
Apoptosis* ; Cornea ; Endothelial Cells ; Endothelium ; Epithelial Cells ; In Situ Nick-End Labeling ; Photorefractive Keratectomy* ; Prognosis ; Rabbits

BACKGROUND: The studies on cryopreserved arterial allograft have been focused on cooling methods, pre-treatment, cryoprotectant agents, and preservation temperature. But recently, several studies have reported that thawing methods also play an important role in the occurrence of macroscopic and microscopic cracks. This study was designed to investigate the cell injury after thawing, using a rabbit model to clarify the effect of thawing methods on cryopreserved arteries. MATERIAL AND METHOD: Segments of the rabbit aorta were obtained and divided into 3 groups (n=60) according to whether the specimens were fresh (control, n=20), cryopreserved and rapidly thawed (RT) at 37oC (n=20), or cryopreserved and subjected to controlled, automated slow thawing (ST)(n=20). Cell damage was established using the TUNEL method and the morphological changes were also evaluated. RESULT: In the group that was rapidly thawed, the expression of TUNEL (+) cells increased significantly more than in the slowly thawed group. In addition, the endothelial denudation, microvesicles and edema were significant in the rapidly thawed group compared with those changes in the slowly thawed group. CONCLUSION: Our study suggests that the rapid thawing method may be one of the major causes of cellular damage and delayed rupture in cryopreserved arterial allografts. The expression of TUNEL (+) cells and structural changes were significantly low in the slowly thawed group, which might have contributed to the improvement of graft failure after transplantation.
Allografts ; Aorta* ; Arteries ; Cryopreservation ; Edema ; In Situ Nick-End Labeling ; Rupture ; Transplants

BACKGROUND: Genistein and daidzein are two major soybean isoflavones. They have received increasing attention because of their possible roles for cancer prevention. However, their mechanisms of action and molecular targets on the human colon cancer cells are not fully understood. METHODS: Human colon cancer HCT-116 cells were treated with genistein and daidzein to investigate their effects on the cell growth and this was analyzed with MTT assay. TUNEL assay and Hoechst33342 stain were carried out to identify apotosis. RESULTS: Daidzein was able to inhibit cell proliferation and induce apoptosis of the HCT-116 cells, but genistein didn't affect the cell growth. The ER antagonist ICI182780 didn't attenuate the antiproliferative and proapoptotic effects of daidzein: this means the effect of daidzein on the HCT-116 cells may not be dependent on the ER pathway. The other soybean isoflavone, genistein, attenuated the effects of daidzein on the HCT-116 cells and its mechanism should be elucidated. CONCLUSIONS: These data suggest that daidzein may act as a preventive agent on human colon cancer, and its mechanism of action doesn't involve the ER-dependent pathway.
Apoptosis ; Cell Proliferation ; Colon* ; Colonic Neoplasms* ; Genistein* ; HCT116 Cells* ; Humans* ; In Situ Nick-End Labeling ; Isoflavones ; Soybeans

PURPOSE: The purposes of this study were to examine whether meniscal degeneration in human osteoarthritis(OA) was related with the occurrence of apoptosis, the expression of nitrotyrosine and Fas. MATERIALS AND METHODS: Menisci were obtained from OA patients undergoing total knee replacement arthroplasty and from normal subjects who were operated an above knee amputaton. According to histologic degeneration, menisci were graded to normal, grade 1(mild), grade 2(moderate), and grade 3(severe). Apoptotic cells were identified by TUNEL method and electron microscopy. Meniscal sections were analyzed by immunohistochemistry for the presence of nitrotyrosine and Fas expression. RESULTS: The number of apoptotic cells were significantly increased in OA meniscus compared with normal meniscus(p < 0.05). The number of apoptotic cells were increased with tissue degeneration. On electron microscopy, the typical chromatin condensation in the OA meniscus was shown in apoptotic cell. The number of Fas-expressing cells was significantly higher in the OA meniscus(p < 0.05). Nitrotyrosine immuno reactivity was prominent in the degenerative menisci(p < 0.05). Fas and nitrotyrosine expression were increased with degree of tissue degeneration. An increase in number of apoptotic cells was correlated with tissue degeneration but not with age . CONCLUSION: Apoptosis was suggested as one of the causes in the tissue degeneration of the human OA meniscus. The development of apoptosis in the meniscus may be related with Fas and nitrotyrosine expression but not with age.
Apoptosis* ; Arthroplasty ; Arthroplasty, Replacement, Knee ; Chromatin ; Humans* ; Immunohistochemistry ; In Situ Nick-End Labeling ; Knee* ; Microscopy, Electron

OBJECTIVE: To investigate the change of placental apoptosis and the expression of their mediator in preeclampsia women. METHODS: Placental tissues from 10 cases of preeclampsia and 15 cases of normal pregnancy were analyzed using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling(TUNEL) staining. Expressions of bcl-2, bax, caspase-3 was also assessed using immunohistochemistry. RESULTS: In TUNEL staining, the number of apoptotic nuclei were significantly increased in the trophoblast of preeclampsia than normal pregnancy. Bcl-2 was mainly expressed in syncytiotrophoblast and bax was mainly expressed in cytotrophoblast. Bcl-2 expression was decreased and bax expression was increased in the preeclampsia than normal, but the difference was not significant. Caspase-3 was mainly expressed in the cytotrophoblast and expression was significantly increased in the preeclampsia than normal pregnancy(p<0.05). CONCLUSION: Placental apoptosis, especially accompanied with increased expression of caspase-3 in cytotrophoblast, might be related with in the pathogenesis of preeclampsia.
Apoptosis* ; Caspase 3 ; Deoxyuridine ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Placenta ; Pre-Eclampsia* ; Pregnancy ; Trophoblasts

PURPOSE: Gene-attenuated replication-competent adenoviruses are emerging as a promising new modality for the treatment of cancer. In an effort to continually improve upon cancer gene therapy, we have modified gene- attenuated replication-competent adenoviruses so as to cause them to replicate efficiently and lyse the infected cancer cells more effectively. MATERIALS AND METHODS: We modified the E1 region of the adenovirus (Ad) systematically, generating Ad-deltaE1B19, Ad-deltaE1B55, Ad-deltaE1B19/55, and Ad-WT. The cytopathic effects (CPE) and viral replication of these four gene modified adenoviruses were compared, and the morphology and DNA fragmentation of the infected cells was evaluated. RESULTS: Among the constructed adenoviruses, E1B 19kD-inactivated adenovirus (Ad-deltaE1B19) was the most potent, inducing the largest-sized plaques and markedCPE. Moreover, cells infected with Ad-deltaE1B19 showed complete cell lysis with disintegrated cellular structure whereas cells infected with Ad-WT maintained intact cellular and nuclear membrane with properly structured organelles. TUNEL assay was also used to monitor DNA integrity, and a more profound induction of apoptosis was observed in the Ad-deltaE1B19 infected cells in comparison to wild type adenovirus infected cells. CONCLUSION: We demonstrate that the inactivation of the E1B19kD gene in a replicating adenovirus leads to increased CPE, rapid viral release, improved cell-to-cell viral spread and increased induction of apoptosis.
Adenoviridae* ; Apoptosis ; Cellular Structures ; DNA ; DNA Fragmentation ; Genes, Neoplasm* ; In Situ Nick-End Labeling ; Nuclear Envelope ; Organelles