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

This experiment was designed to explore the correlation between the mechanism of immobilization-induced skeletal muscle atrophy and the apoptosis of muscular cells. The models of skeletal muscle atrophy induced by immobilization for different length of time were established according to Sievanen II methods. 24 rabbits, each of them having one hind leg fixed by the tubal plaster and the other one free as control, were randomly divided into four groups depending on time of fixation (3, 7, 14, and 28 days respectively). The animals were sacrificed by the end of fixation. TdT-mediated d-UTP nick end labeling (TUNEL) was used to investigate the apoptotic muscle cells in the animal's bone. By comparing the apoptotic muscle cells with the morphology of the skeletal muscle, the correlation between cell apoptosis and skeletal muscle atrophy were analyzed. Apoptotic muscle cells did appear after immobilization in the atrophied skeletal muscle. In various groups, some cells with false positive stained TUNEL were found in the atrophic muscle, which could be distinguished from apoptotic cells by their characteristics. In conclusion, cell apoptosis participates in the process of skeletal muscle atrophy induced by immobilization; the amount of apoptotic cells is strongly associated with the time of immobilization, its peak appears on the 14th day of immobilization; the distribution of apoptotic skeletal muscle cell varies with the time of fixation. The severity of skeletal muscle atrophy is associated with the degree of the muscle cell apoptosis.
Animals ; Apoptosis ; physiology ; Immobilization ; In Situ Nick-End Labeling ; Muscle, Skeletal ; pathology ; Muscular Atrophy ; etiology ; Rabbits

The authorsinvestigated the effects of UV-B and amniotic membrane graft about PRK induced inflammatory cell infiltration into corneal stroma, lipid peroxidation and keratocyte apoptosis. Total 20 white rabbits were divided into 5 groups; 1)mechanical epithelial removal, 2)epithelial removal and UV-B irradiation, 3)PRK only, 4) PRK and UV-B irradiation, 5)Amniotic membrane graft after PRK and UV-B irradiation. All corneas were harvested after 24hrs. H & E stain for PMNs infiltration, MDA immunohistochemical stain for lipid peroxidation and TUNEL stain for keratocyte apoptosis were performed. UV-B had little effect on infiltration of inflammatory cell into corneal stroma, lipid peroxidation and keratocyte apoptosis. Amniotic membrane suppressed infiltration of PMNs into corneal stroma, lipid peroxidation and keratocyte apoptosis. Environmental UV-B exposure should not be avoided after PRK. Amniotic membrane graft is beneficial to reduce keratocyte apoptosis and related corneal haze.
Amnion* ; Apoptosis* ; Cornea ; Corneal Stroma ; In Situ Nick-End Labeling ; Inflammation* ; Lipid Peroxidation* ; Membranes ; Rabbits ; Transplants

PURPOSE: Although immunohistochemically detectable metallothionein (MT) overexpression has been described in proliferation epithelial tumor cells, the clinical significance of the expression remains to be elucidated. Therefore, the present article is focused on evaluating the possible significance of MT expression in colonic adenocarcinoma and its relationship with p53 overexpression, Topoisomerase II-alpha as new cell proliferating marker and apoptosis. METHODS: The following formalin-fixed paraffin embedded surgical or biopsied samples were immunohistochemically stained for MT, p53 and topoisomerase II-alpha, and performed in situ TUNEL method for evaluation of apoptotic cell ; normal control mucosa (78 cases), tubular adenomas (20 cases) and adenocarcinomas with various degree of differentiation (78 cases). RESULTS: The MT immunohistochmical reactivity was decreased in colonic adenocarcinoma than that of normal glandular epithelial and tubular adenoma, with the frequency of MT expression in colonic adenocarcinoma depending upon tumor differentiation only. But the frequency of p53 expression was correlated with T-stage, lymph node metastasis and clinical staging, while topoisomerase II-alpha expression and apoptosis in colonic adenocarcinoma were correlated with lymph node metastasis and clinical staging. The immunohistochemical expression of MT and p53 expression in colonic adenocarcinoma was inversely correlated. Also, the inverse correlation between MT expression and expression of toposiomerase II-alpha indices and apoptotic indices were noted. CONCLUSION: These data suggest that MT expression may play a role in proliferative activity and apoptosis in colonic adenocarcinoma. Although MT expression is correlated to tumor differentiation, further studies of a possibility of prognostic factor, such as p53, are required for the determination of significant relationships in other clinicinopathologic indices.
Adenocarcinoma* ; Adenoma ; Apoptosis* ; Colon* ; In Situ Nick-End Labeling ; Lymph Nodes ; Metallothionein* ; Mucous Membrane ; Neoplasm Metastasis ; Paraffin

PURPOSE: The purpose of this study was to determine whether Fas-L expression is associated with increased apoptotic induction of tumor-infiltrating lymphocytes (TIL) in human gastric carcinomas. MATERIALS AND METHODS: The author analysed 38 cases of early gastric carcinoma (EGC) and 61 cases of advanced gastric carcinoma (AGC) who received gastric resection, in whom the number of diffuse type was 38 cases and the number of intestinal type was 61 cases. The author used immunohistochemical staining for Fas, Fas-L and CD45, and TUNEL in situ apoptosis detection kit. TIL were detected by CD45 and apoptosis of TIL were detected by CD45 expression and TUNEL positivity on serial histologic sections. RESULTS: Fas-L was localized to neoplastic cells in 61% (23/38) of EGC group and 66% (40/61) of AGC group. The extent of Fas-L expression was variable, with both Fas-L positive and negative neoplastic region occuring within tumors. TIL adjacent to Fas-L expressing tumor region were decreased in number and TIL adjacent to FasL-negative tumor region were increased in number; apoptotic induction of TIL showed just the opposite pattern (p<0.05). Fas expression was found essentially homogeneously throughout the tumor mass independent of tumor stage. Fas expression showed 64% (39/61) of intestinal type and 68% (26/38) of diffuse type. Labeling indices for tumoral apoptosis in EGC and AGC were 6.72% and 7.13%, respectively and this difference was statistically insignificant. Co-expression of Fas-L and Fas, which occurred over large areas of the tumors, did not result in an enhanced rate of tumor cell apoptosis. In addition, factors such as tumor stage and other prognostic factors were not concerned in Fas and Fas-L expression, number of TIL and apoptotic induction. CONCLUSION: These findings suggest Fas-mediated apoptotic depletion of TIL in response to Fas-L expression by stomach cancers, and provide the evidence to support the Fas counterattack as a mechanism of immune escape in gastric cancer. In addition, gastric carcinoma cells of the intestinal and diffuse type did not differ in their expression of the apoptotic receptor Fas.
Apoptosis* ; Humans ; In Situ Nick-End Labeling ; Lymphocytes, Tumor-Infiltrating ; Stomach Neoplasms ; United Nations

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

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