ABSTRACT
DcCDPK8 involved in abiotic stress such as low temperature and signal transduction of hormones ABA and MeJA,but the transcriptional regulation is still unclear. In order to study the core promoter region of DcCDPK8 gene in Dendrobium catenatum and explore its transcriptional regulation mechanism,the DcCDPK8 gene promoter sequence was cloned by PCR from D. catenatum. Promoter sequence function was studied by fusion of 5 'terminal deletion and GUS gene. The results showed that the promoter sequence of DcCDPK8 gene has a low-temperature responsive element( LTR) between~(-1) 749 bp and-614 bp,two MeJA responsive elements between~(-1) 749 bp and-230 bp,and one ABA responsive elements between-614 bp and-230 bp. Three 5'-end different deletion fragments were constructed to fuse the eukaryotic expression vectors p BI121 with GUS,which were transformed into tobacco leaves. The GUS activity under cold stress treatment was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3. GUS activity under exogenous ABA induction was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3,and GUS activity under exogenous MeJA induction was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3. It is speculated that the ABA response element( ARE) in the promoter sequences of DcCDPK8 is positive regulatory role in response to exogenous ABA,the MeJA cis-acting element plays a negative role in response to exogenous MeJA.
Subject(s)
Abscisic Acid , Acetates , Cloning, Molecular , Cold Temperature , Cyclopentanes , Dendrobium , Genetics , Gene Expression Regulation, Plant , Oxylipins , Plant Proteins , Genetics , Plants, Genetically Modified , Promoter Regions, Genetic , Response Elements , Stress, Physiological , NicotianaABSTRACT
Objective To explore the cell viability in the ablation area of thyroid nodules at 6 months after microwave ablation by enzyme histochemical staining. Methods Twenty-four ablation areas of thyroid nodules were selected from 20 patients who underwent histopathological assessment of the ablation area by core needle biopsy at 6 months after microwave ablation between Dec. 2017 and Sep. 2018. Core needle biopsy was performed at the central and marginal regions of the ablation area with a cutting biopsy needle. The specimens were obtained and placed in liquid nitrogen to make frozen sections. Enzyme histochemical staining was used to detect the activities of succinate dehydrogenase (SDH) and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d), and the difference of cell morphology and histological structure was compared with H-E staining results. Results The specimens of the central and marginal regions of 24 ablation areas were successfully obtained. The histochemical staining of SDH and NADPH-d in the central region of ablation area had good consistency, and the negative rates were both 95.83% (23/24). The histochemical staining of SDH and NADPH-d in the marginal region of ablation area also had good consistency, and the negative rates were both 91.67% (22/24). H-E staining of 23 central regions and 22 marginal regions showed pink amorphous mass of necrosis. H-E staining of 1 central region and 2 marginal regions showed partly necrotic and fibrous tissue hyperplasia. The location of fibrous tissue hyperplasia was consistent with the location of the positive region of enzyme histochemical staining. Conclusion At 6 months after microwave ablation, the tissue in the ablation area of thyroid nodules is consistent with coagulative necrosis, and is still inactivated. SDH or NADPH-d enzyme histochemical staining combined with H-E staining can objectively evaluate the old ablation area.
ABSTRACT
To determine the prevalence of goblet cell metaplasia in endocervical and endometrial adenocarcinomas by histochemial staining and to investigate the most sensitive histochemical staining method to detect this metaplasia, a total of 90 tissue blocks representing 30 non-neoplastic cervix, 30 non-neoplastic endometrium, 30 endocervical and endometrial adenocarcinoma cases were obtained for histochemical staining with Toluidine Blue (TB), Methylene Blue (MB), Mucicarmine (MUC), Periodic Acid Schiff before and after Diastase digestion (PAS, PAS-D), Alcian Blue pH 2.5 (AB), and Periodic Acid Schiff after Alcian Blue pH 2.5 (PAB). The cases were blinded and evaluated by a pathologist [NHO] for the presence of goblet cell metaplasia, the amount of goblet cells present and the histochemical differentiation of the goblet cells compared with its surrounding glandular epithelium. Goblet cell metaplasia was present in 2 out of 30 cases in non-neoplastic cervix, 0 out of 30 cases in non-neoplastic endometrium, 7 out of 15 cases in endocervical adenocarcinoma and in 2 out of 15 cases in endometrial adenocarcinoma. Relatively few goblet cells were seen in endometrial adenocarcinoma, few to moderate amounts were seen in endocervical adenocarcinoma and relatively more goblet cells were seen in non-neoplastic cervix. The differentiation of the goblet cells with its surrounding glandular epithelium was moderate to strong in non-neoplastic cervix and endocervical adenocarcinoma, while the differentiation in endometrial adenocarcinoma was weak to moderate. The various staining methods showed differences in presence, amount anddifferentiation of the goblet cells. Goblet cell metaplasia of the reproductive organs is not as rare as previously reported. There was no statistical difference in presence, amount and differentiation of goblet cells according to the various cases. The must optimum staining methods for staining goblet cells in non-neoplastic cervix, endocervical adenocarcinoma and endometrial adenocarcinoma were PAS, PASD and AB.
ABSTRACT
Cutaneous tuberculosis presents a diagnostic challenge, as it is difficult to demonstrate the causative organism by histopathology and also culture of organisms from skin lesions is a less rewarding and time consuming process. Aim: Present study was undertaken to evaluate the utility of immuno-histochemical staining to demonstrate Mycobacterium tuberculosis antigen in tissue sections. This is based on the finding that the mycobacterial antigen is the last to disappear from the tissues and thus can be used as a marker of mycobacterial infections. Material & Methods: Fifty randomly selected skin biopsy specimens were subjected to routine histopathological examination to corroborate the clinical diagnosis. Immuno-histochemical study was undertaken to demonstrate mycobacterial antigen. Observations: All the tissue sections were negative for AFB, both by Z-N stain and by culture. Mycobacterial antigen was demonstrable in 68% of cases of cutaneous tuberculosis. The highest positivity was recorded in scrofuloderma (89%), followed by Lupus Vulgaris (69%) and Tuberculosis Verrucosa Cutis (47%). Conclusions: Mycobacterial antigen was demonstrable in majority of cases of cutaneous tuberculosis using polyclonal antiserum. However, since cross reactivity was reported in cases of leprosy and also in some fungal infections, this test by itself cannot be considered as diagnostic. The results should be considered along with other findings.
ABSTRACT
Objective;To investigate whether the traditional Chinese medicine Yisheng Injection can reduce the injury on isolated rat heart induced by cold ischemia. Methods ;18 isolated female closed colony SD rats were divided into two groups at random. In the control group, the isolated rat hearts were preserved in 0.9% sodium chlorine solution at 4℃ , while in the experimental group ,the hearts were preserved in 0.9% sodium chlorine solution containing Yisheng injection at 4℃. The specimens were harvested 2,6,12, 24,48 and 72 hours after preservation. Histochemical changes were observed in two groups. Results ;In the control group, the activities of alkaline phosphatase ( ALP) and NADPH diaphorase (NADPHD) decreased after cold ischemia for 24 hours. Lactic dehydro-genase (LDH) decreased slightly, while ALP and NADPHD decreased evidently, after cold ischemia for 48 hours. Succinic dehydro-genase ( SDH) and cytochrome oxidase ( CCO) decreased slightly, and NADPHD disappeared after cold ischemia for 72 hours respectively. In the experimental group, NADPHD, LDH and CCO decreased slightly after cold ischemia for 24, 48 and 72 hours. ALP decreased slightly after cold ischemia for 48 hours, however, SDH showed no changes, after cold ischemia for 72 hours. Conclusion : Cold ischemia can induce cold ischemia injury in isolated rat hearts, especially the vascular endothelial cells being more sensitive. Yisheng injection can reduce this kind of injury to some degree.
ABSTRACT
Objective:To investigate the injury on isolated cadaver testes induced by ischemia-reperfusion (I/R).Methods:13 isolated cadaver testes contributed by 13 persons were preserved under 0℃～4℃ hypothermia and then reperfused under 37℃.Histology and histochemical changes were observed.Results:4℃ cold ischemia in 18 hours only could induce trivial swelling and vascular degeneration of endothelial cells(ECs).Obvious pathologic changes occurred after 24 hours,including detachment of ECs,separation between basement membrane and seminiferous epitelium,malalignment of spermatogenous cell and edema of mesenchyme.Injury was worse along with the prolongation of cold preservation time.Changes of LDH and SDH activities were found by histochemical staining.Reperfusion following 6 hours ischemia induced tissue injury and unusual enzyme activity.All changes were more obvious after reperfusion following 12,18,24 or 36 hours of cold ischemia.Conclusion:HCA can preserve testes for 18 hours.Testes can't keep normal histological and histochemical structure beyond 24 hours of preservation under 4℃ hypothermia,and 37℃ reperfusion can make injury worse.
ABSTRACT
Objective Peritoneal lymphatic features are studied to provide necessary data for understanding transport capabilities of the endothelial and mesothelial cells. Methods Enzyme\|histochemical staining methods were employed to investigate organization of the lymphatic networks and their endothelial ultrastructures in the monkey peritoneum using light,scanning and transmission electron microscopy. Results 5' Nase positive initial lymphatics showed extensive network,obvious valve like structures and numerous blind ends.The calibre of lymphatics with extremely irregular lumen varied greatly from 40 to 120?m.Lymphatic endothelium was usually separated from mesothelium by a small quantity of loose connective tissue,or they directly contacted each other in some areas.No basal lamina occurred between the peritoneum and the lymphatics originating from milky spots in the omentum and mesovarium.Milky spots are oval or round visible bodies aggregated by macrophages and lymphocytes.Both the endothelium of lymphatic lacunae and the mesothelium that forms the peritoneal stomata,represent a strong 5' Nase activity.Abundant microfibrils attached the stomatal edge. Conclusion The distribution and structure of peritoneal initial lymphatics reveal significant regional variations,and lymphatic endothelium has a close morphological and functional relationship with the mesothelium. [