Abnormal change of calcium signal system on consecutive monoculture problem of Rehmannia glutinosa / 中国中药杂志

Based on the early transcriptome and digital differentially expressed profiling library construction in consecutive monoculture (two-year culturing) Rehmannia glutinosa, we screened and chose the twelve differentially expressed protein genes which might be related with calcium signal system. The spatiotemporal expression of these genes was measured by the real-time quantitative PCR, and the relative expression values of the genes related with calcium signal system in different development stages and tissues of normal growth (one-year culturing) and succession cropping of R. glutinosa (two-year culturing) was elaborated in detail. In addition, disposed succession cropping of R. glutinosa was treated with different levels of calcium signal blocking agents in order to verify the mode of action of calcium signal system on consecutive monoculture problem in R. glutinosa. Among the twelve genes, two calcium channels away from the cytoplasm were down-regulated expressed, while the ten calcium channels toward the cytoplasm were up-regulated expressed. The results implied that succession cropping caused calcium ions flowing from endoplasmic reticulum to cytoplasm. While the key genes in calcium signal respond components such as CBL, CBP, CIBP, PLC, etc. were down-regulated expressed significantly in succession cropping of R. glutinosa which were disposed with calcium signal blocking agents, the extent of the damage was relieved, and approached the normal growth (one-year culturing) level. This result strongly showed that calcium signal system participated in the perceiving, conducting and magnifying processes of succession cropping obstacles of R. glutinosa.

Evaluation of the expression and significance of Claudin-5 and CD99 in solid-pseudopapillary neoplasms and neuroendocrine tumors of pancreas / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of endothelium tight junction protein Claudin-5 and intercellular adhesion molecule CD99 in solid-pseudopapillary neoplasms (SPN) and neuroendocrine tumors of pancreas (P-NET), and their significance in the differential diagnoses.</p><p><b>METHODS</b>Immunohistochemical staining of Claudin-5 and CD99 was performed in 37 cases SPN and 21 cases of P-NET.</p><p><b>RESULTS</b>Membranous Claudin-5 expression was observed in all cases of SPN but was absent in all cases of P-NET. The difference was significant (P < 0.01). In SPN, 91.9% (34/37) of the cases displayed paranuclear dot-like immunoreactivity for CD99; in contrast, 61.9% (13/21) of the cases of P-NET displayed membranous staining (P < 0.01). There was a positive association between the expression of Claudin-5 and CD99 in SPN (r = 0.421,P = 0.001).</p><p><b>CONCLUSIONS</b>Although the macroscopic and microscopic features of SPN are quite characteristic, they may not allow confident differentiation from P-NET in all cases, especially when these characteristics are not classical. If necessary, immunostaining for Claudin-5 and CD99 can help to differentiate between these entities.</p>

Spatiotemporal expression and analysis of responding consecutive monoculture genes in Rehmannia glutinosa / 中国中药杂志

<p><b>OBJECTIVE</b>Based on previous study, authors used the suppression subtractive hybridization (SSH) technique to construct the forward and reverse subtractive cDNA libraries of consecutive monoulture Rehmannia glutinosa. Five genes related with consecutive monoculture problem of R. glutinosa were chosen from the each of two subtractive libraries. And their spatiotemporal expression was measured in order to explore the functions in consecutive monoculture problem of R. glutinosa.</p><p><b>METHOD</b>Using the real-time quantitative PCR, we tested the relative expression values of the genes in different development stages and tissues of normal growth (one-year culturing) and consecutive monoculture (two-year culturing) R. glutinosa.</p><p><b>RESULT</b>The five genes (calcium-dependent protein kinase, s-adenosyl-methionine synthetase, Aminocyclopropane-1-carboxylate oxidase, methyltransferase, calpain), which were chosen from the forward library had high expression in consecutive monoculture R. glutinosa, especially in root, and were hardly expression in normal growth R. glutinosa. On the contrary, the other five genes (RNA-dependent RNA polymerase, RNA replicase, DNA-directed RNA polymerase IIa, cyclin D, RNA binding protein) chosen from the reverse library had high expression in one-year R. glutinosa, but were down regulated or shut down in consecutive monoculture R. glutinosa.</p><p><b>CONCLUSION</b>The key genes, which regulate inessential metabolism parthway (such as cyclin D, DNA-directed RNA polymerase IIa), were restrained or shut down in consecutive monoculture R. glutinosa. Calcium and ethylene signaling might played key roles in the formation of consecutive monoculture problem, resulting in disturbing normal metabolic process and syndrome of disease in R. glutinosa appeared in turn.</p>