New Oral Agent for Treatment of Anemia in Patient with Chronic Kidney Disease: Prolyl Hydroxylase Inhibitor / 대한내과학회지

Hypoxia inducible factor (HIF)-stabilizers are being developed for the renal anemia treatment. This small molecules inhibit prolyl hydroxylase domain (PHD)-containing enzymes, causing HIF activation instead of degradation under the state of normoxia, finally increase production of intrinsic erythropoiesis. Current treatment guidelines suggest that renal anemia should be treated mainly with iron and erythropoiesis stimulating agents (ESAs). But there are several complications and concerns such as hypertension, ESA refractory anemia and increased cardiovascular mortality in using ESAs. Advantages of HIF stabilizers over ESAs are orally available, no dose-up requirement for inflammation. So far new HIF stabilizers showed efficacy and safety in renal anemia treatment. This new therapeutic agent may emerge as a standard treatment option for renal anmia treatment.

Isolation of cancer stem cells from pancreatic cancer cell line and identification of their biological characteristics / 中国药理学通报

Aim To isolate cancer stem cells from human pan-creatic cancer cell line L3. 6pl and to identify their biological characteristics. Methods L3. 6pl cells were cultivated in com-mercial low adhesion plate with serum-free stem cell culture me-dium ( MEM/F12 1:1 ) supplemented with B27. The cancer stem cells reformed into floating spheres were isolated. The method of tumor sphere formation was used to isolate/enrich and characterize the cancer stem cells in pancreatic carcinoma cell line L3. 6pl. Cancer stem cell spheres were collected and sorted using magnetic cell sorting ( magnetic activated cell sorting, MACS) technology, with the cell surface markers of CD24 +CD44 + ESA+. Self-renewal and EMT-related oncogene expres-sion were measured with Western blot. Cancer stem cells differ-entiation potential and the expression of cancer stem cell related signs were checked with Immunofluorescence assay. To deter-mine tumorigenesis in vivo, Xenograft assay in NOD-SCID mice were performed respectively, then immunohistochemistry proto-oncogene c-Met and RON expression were also checked. West-ern blot was used to detect the changes of stemness relative tran-scriptional factors and epithelial markers expressed in spheres before and after differentiation. Drug resistance of pancreatic cancer stem cells to gemcitabine or paclitaxel was measured with MTT assay. Results CD24 + CD44 + ESA+ cells were signifi-cantly tumorigenic, and cultured in serum-free conditions to form spheroids, which had the characteristics of stem cells with self-renewal, EMT and drug-resistant capabilities, and had a posi-tive correlation with the c-Met, RON protein expression. Con-clusion Human pancreatic stem cells are successfully isolated, which provides a useful model for individualized therapy and e-valuation of the therapeutic efficacy for pancreatic cancer pa-tients.

High diagnostic efficiency of IgM-ELISA with the use of multiple antigen peptides (MAP1) from T. gondii ESA (SAG-1, GRA-1 and GRA-7), in acute toxoplasmosis / Alta eficiência diagnóstica do IgM ELISA com o uso de múltiplos antígenos peptídicos (MAP1) de T. gondii ESA (SAG-1, GRA-1 e GRA-7) na toxoplasmose aguda

The main serological marker for the diagnosis of recent toxoplasmosis is the specific IgM antibody, along with IgG antibodies of low avidity. However, in some patients these antibodies may persist long after the acute/recent phase, contributing to misdiagnosis in suspected cases of toxoplasmosis. In the present study, the diagnostic efficiency of ELISA was evaluated, with the use of peptides derived from T. gondii ESA antigens, named SAG-1, GRA-1 and GRA-7. In the assay referred to, we studied each of these peptides individually, as well as in four different combinations, as Multiple Antigen Peptides (MAP), aiming to establish a reliable profile for the acute/recent toxoplasmosis with only one patient serum sample. The diagnostic performance of the assay using MAP1, with the combination of SAG-1, GRA-1 and GRA-7 peptides, demonstrated better discrimination of the acute/recent phase from non acute/recent phase of toxoplasmosis. Our results show that IgM antibodies to MAP1 may be useful as a serological marker, enhancing the diagnostic efficiency of the assay for acute/recent phase of toxoplasmosis.

O principal marcador sorológico para o diagnóstico da toxoplasmose aguda ou recente são os anticorpos IgM específicos, a par de anticorpos IgG de baixa avidez. Entretanto em alguns pacientes, estes anticorpos podem persistir, ultrapassando o período da fase aguda/recente, contribuindo para erro diagnóstico em casos suspeitos de toxoplasmose. No presente estudo, a eficiência diagnóstica de ELISA foi avaliada, com o uso de frações ou peptídeos originados dos antígenos ESA de T. gondii, denominados de SAG-1, GRA-1 e GRA-7. No referido ensaio, estudamos cada uma destas frações isoladamente, como também em quatro diferentes combinações, ou múltiplos peptídeos antigênicos (MAP), visando estabelecer um perfil confiável para a toxoplasmose aguda/recente em amostra única de soro. A melhor eficiência diagnóstica do ensaio foi encontrada com o uso da combinação de peptídeos SAG-1, GRA-1 e GRA-7, denominada MAP1. A detecção de anticorpos IgG e IgM anti-MAP-1 apresentou melhores características diagnósticas para a diferenciação entre a fase aguda/recente da fase não aguda/recente na toxoplasmose. Nossos resultados mostram que anticorpos IgM anti-MAP-1 poderão prestar auxílio como um marcador sorológico, aumentando a eficiência diagnóstica do ensaio para a fase aguda/recente da toxoplasmose.

Studies on Specific Diagnostic Antigens in Excretory-secretory Products from Trichinella spiralis Muscle Larvae / 中国寄生虫学与寄生虫病杂志

Objective To find out the specific diagnostic antigens in excretory-secretory (ES) products from muscle larvae of Trichinella spiralis. Methods The ES antigens (ESA) of Trichinella spiralis muscle larvae cultured in vitro at 18 h and 30 h were analyzed by SDS-PAGE and Western blotting. Results At different times after cultivation, the protein components of ESA of T. spiralis muscle larvae were similar. SDS-PAGE revealed that the molecular weight(MW) of the major bands of 2 ES antigens were 112, 110, 108, 97, 53, 49, 45, 42, 35, 23 and 16 kDa. Western blotting showed that the protein bands with 102, 97, 95 and 53 kDa in 18 h ESA and the protein bands with 53, 49, 45 and 43 kDa in 30 h ESA cross-reacted with sera from the patients with paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis, respectively. The protein component with 23 kDa in ESA only reacted with sera from the rats and mice infected with T. spiralis and the patients with trichinellosis, but not reacted with sera from animals and patients infected with other parasites, and sera from normal rats, mice and persons. Conclusion The protein component with 23 kDa in T. spiralis ESA is the specific antigen of T. spiralis muscle larvae and it could be applied to the serodiagnosis and seroepidemiological survey of trichinellosis.

Effects of ethanol sediments from tuber of Angelica sinensis and its litmusless component on secreting TNF-? and IL-1 of macrophages in vitro / 中草药

Object To investigate the effects of ethanol sediments obtained from the tuber of Angelica sinensis (Oliv). Diels (ESA) and its litmusless component (ESA 1) on the secretion of TNF ? and IL 1 by mice peritoneal macrophages in vitro. Methods L929 cell line cytotoxicity was used for the assay of TNF ?. The proliferation of L929 cell line was used for the assay of IL 1. Results The secretion of TNF ? and IL 1 by mice peritoneal macrophages which were co cultured with ESA or ESA 1 in vitro can be significantly promoted. At the concentrations in range of 5～20 ?g/mL, there is a dose dependence in the action of ESA, while there is not the similar effect of ESA 1, even though it showed the marked effect. Conclusion ESA and ESA 1 can enhance the secreting TNF ? and IL 1 of mice peritoneal macrophages in vitro.