Inhibition of Sonic Hedgehog signaling inhibits fibrous scar formation and adversely affects functional outcome after ischemic brain injury in rats / 南方医科大学学报

OBJECTIVE@#To investigate the effects of inhibiting Sonic Hedgehog (Shh) signaling on fibrous scar formation and functional outcome after ischemic brain injury.@*METHODS@#Adult SD rats were randomized into sham-operated group, middle cerebral artery occlusion (MCAO) and reperfusion (I/R) group, I/R with intraventricular empty adenoviral vector (rAd-NC) injection group, and I/R with adenovirus-mediated Shh knockdown (rAd-ShShh) group. After the treatments, the neurological deficits of the rats were assessed, and the protein and mRNA expressions of fibronectin (Fn), α-SMA, and Shh in the ischemic hemisphere were detected with immunofluorescence assay and qPCR; TUNEL staining was used for detecting neural cell apoptosis. In the cell experiment, primary meningeal fibroblasts isolated from neonatal SD rats were pretreated for 24 h with TGF-β1 or TGF-β1 plus cyclopamine (CYC) before oxygen-glucose deprivation for 150 min followed by reoxygenation for 72 h (OGD/R). CCK-8 assay and scratch test were performed to examine the changes in cell proliferation and migration, and immunofluorescence assay, qPCR and Western blotting were used for detecting cell transformation and the expressions of Shh, α-SMA, and Fn.@*RESULTS@#Cerebral I/R injury significantly increased the protein and mRNA expressions of Shh, α-SMA, and Fn in the ischemic hemisphere of the rats, but their expression levels were significantly lowered by intraventricular injection of rAd-Shshh (P < 0.05), which obviously increased cell apoptosis in the ischemic hemisphere (P < 0.05) and improved modified mNSS and modified Bederson scores of the rats (P < 0.05). In the cell experiment, pretreatment with TGF-β1 and TGF-β1+CYC both increased the viability of the primary meningeal fibroblasts after OGD/R. TGF-β1 significantly enhanced the migration ability and induced obvious transformation of the exposed cells (P < 0.05), but these effects were significantly attenuated by co-treatment with CYC (P < 0.05). The expressions of Shh, α-SMA and Fn in the TGF-β1 group were all significantly higher in TGF-β1-treated cells (P < 0.05) and were obviously lowered by co-treatment with CYC (P < 0.05).@*CONCLUSION@#Inhibition of Shh signaling may inhibit fibrous scar formation and functional recovery in rats after ischemic brain injury.

Consenso de leucemia mieloide aguda en México / Mexican consensus in acute myeloid leukemia in Mexico

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Abstract Acute myeloid leukemia (AML) comprises a heterogeneous group of hematopoietic cell neoplasms of myeloid lineage that arise from the clonal expansion of their precursors in the bone marrow, interfering with cell differentiation, leading to a syndrome of bone marrow failure. AML is a consequence of genetic and epigenetic changes (point mutations, gene rearrangements, deletions, amplifications, and arrangements in epigenetic changes that influence gene expression) in hematopoietic precursor cells, which create a clone of abnormal cells that are capable of proliferating but cannot differentiate into mature hematopoietic cells or undergo programmed cell death. The diagnosis requires more than 20% myeloid blasts in the bone marrow and certain cytogenic abnormalities. Treatment will depend on age, comorbidities, and cytogenetic risk among the most frequent.

Analysis on clinical efficacy and influencing factors of decitabine-based regimens in patients with myelodysplastic syndrome-refractory anemia with excess blasts / 中国医师杂志

Objective To analyze the clinical efficacy,safety and influencing factors of decitabine (DAC)-based regimens in patients with myelodysplastic syndrome-refractory anemia with excess blasts (MDS-RAEB).Methods We performed a retrospective analysis of 63 patients with MDS-RAEB treated with DAC,evaluated the clinical efficacy and adverse reactions,and analyzed the influencing factors affecting survival.Results Among 63 patients,23 were RAEB-1 and 40 were RAEB-2.The median treatment was 4 (2-13) courses.The total effective rate of DAC for MDS-RAEB was 58.7％ (37/63),and the complete response rate was 20.6％ (13/63).Among 37 patients who were effective,20 (54.1％) patients performed efficacy after 2 courses.The median course of treatment to achieve the best effect was 3.5 (3-4) courses.With a median follow-up of 14 (2-68) months,63 patients had a overall survival rate (OS) of 84.2％ and a 1-year progression-free survival rate (PFS) of 73％.In univariate analysis,the factors that prolonged OS were that the best effect after medication was stable disease (SD) (to achieve complete remission,partial remission,complete bone marrow remission,hematological improvement) (P =0.009) and no thrombocytopenia at first diagnosis (P =0.019),the factor that prolongs PFS is the best effect above SD (P =0.003).Multivariate analysis suggested that the factors affecting OS and PFS were the best curative effects above SD (P =0.015 vs P =0.008).The adverse effects of decitabine in the treatment of MDSRAEB were mainly bone marrow suppression and pulmonary infection.Conclusions Decitabine is an effective and well-tolerated drug in the treatment of MDS-RAEB.Response to decitabine treatment is one of the independent factors affecting the prognosis.

Is this a blast? An illustrated practical review on peripheral blood smear examination in the paediatric patient

@#The morphologic findings on a peripheral blood smear can provide important clues that help establish a diagnosis or guide the workup of many clinical disorders. Finding a blast – whether clinically expected or not – is one of the most impactful of such findings. Pathologists, clinical haematologists, technologists, and trainees in the medical field often feel the need to refer to an illustrated reference when encountering suspected blasts and blast-mimics. This article provides a practical concise resource that demonstrates the morphological features of the various types of blasts and illustrates the cytologic characteristics that help distinguish them from their benign mimickers in the paediatric population.

Effect of mir-152 on proliferation of myocardial fibroblasts in diabetic cardiomyopathy / 中国药理学通报

Aim To explore the effect of miR-152 on proliferation of cardiac fibroblasts ( CFS) in diabetic cardiomyopathy. Methods Diabetic cardiomyopathy model was established in SD rats by STZ injection, and CFS proliferation model was established by high glucose (33. 3 mmol • L ~1). HE and Masson staining were performed in paraformaldehyde fixed myocardium of rats. Western blot determined a-SMA and collagen I protein expression. qPCK detected gene expression of miR-152. MTT assay analyzed the proliferation of cells. Results HE and Masson staining showed the higher level of myocardial collagen in diabetic cardiomyopathy model. Furthermore, the myocardial myo-cytes lined up in disorder. Western blot showed that the expressions of a-SMA and collagen I were up-regulated in the diabetes mellitus ( DM ) group, while the expression of miR-152 was down-regulated. The result of the in vitro experiment showed that a-SMA and collagen I expressions were down-regulated after trans-fected miR-152 mimics. The proliferation of CFS was also down-regulated after transfected miR-152 mimics. Conclusions miR-152 plays an important role in the proliferation of CFS and may ameliorate diabetic cardiomyopathy.

Soluble HLA-G: A novel marker in acute myeloid leukemia patients

Objective: to study soluble HLA-G (sHLA-G) in Egyptian acute myeloid leukemia (AML) patients. HLA-G is speculated to be a tumor-driven immune escape mechanism. In addition, it might be a promising target for future immune therapeutic approaches. Methods: Thirty AML patients and 15 healthy controls of matched age and sex were the subject of the study. sHLA-G was done to all patients and controls by ELISA. Results: Statistically significant increase in sHLA-G level was present in AML patients compared to controls, being higher in relapsed cases. HLA-G levels was correlated to bone marrow blast percentages but not affected by age, gender, WBCs or response to chemotherapy. HLA-G had a sensitivity of 100% and a specificity of 62% to detect AML cases. Conclusion: HLA-G may be an additional marker for AML especially relapsed cases

Research progress on regulation of angiotensin Ⅱ on bone metabolism / 中国药理学通报

The renin-angiotensin system exists locally in bone tissue.The RAS components and the active peptide angiotensinⅡ (Ang Ⅱ)within RAS are directly involved in the pathophysi-ological process of regulating bone metabolism.Ang Ⅱ exerts the effects via its receptors expressed in osteoblasts and osteoclasts. This paper reviewed the pathways of classical RAS,explained the tissue RAS-induced tissue injuries,and especially elaborated the regulation of RAS active peptide Ang Ⅱ on target genes and in-tracellular signaling pathways in osteoblasts,the action of Ang Ⅱon osteoclastic function via acting on osteoblasts,and the inter-action of Ang Ⅱ receptors involved in the modulation on bone metabolism.

Inhibitory effects of resveratrol on human pulmonary fibroblast line MRC-5 growth / 中国药理学通报

Aim To explore the inhibitory effects of resveratrol (Res ) on human pulmonary fibroblast growth,and its related mechanisms.Methods Hu-man pulmonary fibroblasts MRC-5 were cultured in vitro as research object.These cells were inoculated with 20 μL dimethyl sulfoxide (DMSO)as well as 0, 12.5,25 50,100 and 200 μmol·L-1 Res for 24,48 and 72 h,respectively.Inhibitory rate of cellular pro-liferation was analyzed by MTT.In addition,these cells were treated with 20 μL DMSO (medium group) as well as 50 and 100 μmol·L-1 Res for 48 h,re-spectively.Subsequently,cell cycle and apoptotic rate were measured using flow cytometry.Apoptosis index (AI ) was detected through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The mRNA and protein expression of cell cycle protein D1 (Cyclin D1 ) and cyclin-dependent kinase 4 (CDK4)was detected through fluorescence real-time quantitative PCR and Western blot, respectively. Western blot was used to measure the protein expres-sion of Bcl-2 and Bax.Results With the increase of Res concentrations and prolongation of treated time, inhibitory rate of cellular proliferation was gradually el-evated (P<0.01).After 48 h of co-culture,DNA ra-tio of S and G2/M periods,mRNA and protein levels of Cyclin D1 and CDK4,and Bcl-2 protein levels were significantly decreased while DNA ratio of G0/G1 peri-od,AI,apoptotic rate and Bax protein levels were sig-nificantly increased in 50 and 100 μmol · L-1 Res-treated groups as compared to medium group (P <0.01 ).Moreover,the effects 100 μmol · L-1 Res were better than those of 50 μmol · L-1 Res (P <0.01).Conclusion Res can suppress the prolifera-tion of MRC-5 cells,which may be associated with blockade of cell cycle progression and induction of cell apoptosis.

Flow Cytometric White Blood Cell Differential Using CytoDiff is Excellent for Counting Blasts

BACKGROUND: The usefulness of the CytoDiff flow cytometric system (Beckman Coulter, USA) has been studied in various conditions, but its performance including rapidity in detecting and counting blasts, the most significant abnormal cells in the peripheral blood, has not been well evaluated. The objective of this study was to evaluate the performance of the CytoDiff differential counting method in challenging samples with blasts. METHODS: In total, 815 blood samples were analyzed. Samples flagged as "blasts" or "variant lymphocytes" and showing 0.8) for neutrophils and lymphocytes but poor (r<0.8) for other cells. When the cutoff value of the CytoDiff blast count was set at 1%, the sensitivity was 94.4% (95% CI; 91.2-96.6) and specificity was 91.9% (95% CI; 89.0-94.1). The positive predictive value was 88.4% (95% CI; 84.4-91.5) (304/344 cases) and negative predictive value was 96.2% (95% CI; 93.9-97.7) (453/471 cases). The CytoDiff blast counts correlated well to the manual counts (r=0.9223). CONCLUSIONS: The CytoDiff method is a specific, sensitive, and rapid method for counting blasts. A cutoff value of 1% of at least 1 type of blast is recommended for positive CytoDiff blast counts.

Perfil sociodemográfico e clínico de portadores de síndromes mielodisplásicas / Sociodemographic and clinical profile of patients with myelodysplastic syndromes

Estudos epidemiológicos sobre Síndromes Mielodisplásicas (SMD) não são encontrados na literatura brasileira, o que requer investigação dessa doença prevalente em idosos e com incidência maior com o aumento da idade. Esse trabalho objetivou investigar o perfil sociodemográfico e clínico dos pacientes portadores de SMD. Tratase de um corte transversal, desenvolvido no Rio Grande do Norte, realizado de janeiro de 2000 a dezembro de 2010. Para análise descritiva foi utilizado o programa Epi Info 2002, versão 3.5.2. Os cálculos da probabilidade de associação entre as características analisadas e o gênero foram realizados pelos Testes do qui-quadrado, de Fisher e Exato de Fisher. O nível de significância considerado foi de 0,05. O trabalho foi aprovado em seus aspectos éticos e metodológico pelo Comitê de Ética em Pesquisa CEP/HUOL protocolo 432/10. Dos 29 pacientes selecionados, houve predomínio de idosos, do sexo masculino, com baixa escolaridade, que apresentaram anemia como sintoma inicial. A maior parte foi de pessoas de pele branca, residentes em casa própria, moradores em zona urbana e com renda inferior a dois salários mínimos. Todos utilizaram terapia com hemoderivados, principalmente o concentrado de hemácias, numa frequência de quatro ou mais unidades por mês de consumo, sendo que 20% realizou dosagem de ferritina sérica, todos com valores acima do normal referenciado. Conclui-se que se faz necessário a realização de pesquisas com maiores populações, de caráter multicêntrico a fim de melhor evidenciamento dos dados sociodemográficos e clínicos com possibilidade de avaliação por regiões do país.

Epidemiological studies on Myelodysplastic Syndromes (MDS) are not found in Brazilian literature, which requires investigation of this prevalent disease in the elderly and higher incidence with increasing age. This study aimed to investigate the sociodemographic and clinical characteristics of patients with MDS to characterize this population at a referral center for high complexity. It is a cross-performed from January 2000 to December 2010. For descriptive analysis was conducted using Epi Info 2002, version 3.5.2. The calculations of the likelihood of association between the characteristics analyzed and gender were performed using the chi-square, Fisher and Fisher's Exact. The level of significance was 0.05. The study was approved in its ethical aspects and the methodological Ethics Committee in Research ECR/HUOL Protocol 432/10. We selected 29 patients. The sample was characterized mainly by elderly male with lower education, who had anemia as initial symptom. Most were white-skinned people living in their own homes, residents in urban areas and with income less than two minimum wages. All blood products used therapy, especially red blood cells, a frequency of four or more units per month of consumption. Only 20% performed dosage of serum ferritin, all with values referenced above normal. We conclude that it is necessary to conduct research with larger populations, multicenter character in order to best evidence on the demographic data and clinical evaluation with the possibility of the country.

Sobrevida e evolução leucêmica de portadores de síndromes mielodisplásicas / Survival and leukemic evolution of patients with myelodysplastic syndromes

As síndromes mielodisplásicas constituem um grupo de doenças de ordem clonal hematopoética evidenciadas por estudos em todo o mundo. A estimativa de sobrevida dos pacientes e de casos que apresentam evolução leucêmica requer investigação na população brasileira, pois não se conta com nenhum dado dessa natureza. Assim, este estudo objetivou caracterizar e estimar a sobrevida e evolução leucêmica de portadores de síndromes mielodisplásicas acompanhados em um serviço de referência. Trata-se de um estudo de coorte retrospectiva realizada de janeiro de 2000 a dezembro de 2010. Para análise descritiva foi utilizado o programa Epi Info 2002, versão 3.5.2, e para os cálculos das probabilidades de sobrevida foi empregado o método de Kaplan-Meier pelo uso do programa Statistic Package for Social Sciences (SPSS), versão 10.0. Os cálculos da probabilidade de associação entre as características analisadas e o gênero foram realizados pelos testes do qui-quadrado de tendência, de Fisher, Mann Whitney, e de Log Rank. O nível de significância considerado foi de 0,05. O trabalho foi aprovado em seus aspectos ético e metodológico pelo Comitê de Ética em Pesquisa do Hospital Universitário Onofre Lopes (HUOL), sob o Protocolo n. 432/10. Dos 29 pacientes selecionados, houve predomínio de idosos, do sexo masculino, com baixa escolaridade. Apresentaram baixa probabilidade acumulada de sobrevida e índices de evolução leucêmica em torno de 27%, sem nenhum resultado satisfatório para o tratamento quimioterápico, bem como nenhuma indicação de transplante de medula óssea como possibilidade de cura. Fazem-se necessárias pesquisas com populações maiores para caracterização em todo território nacional.

Myelodysplastic syndromes constitute a group of clonal hematopoietic diseases shown by studies all around the world. The survival estimation of the patients and the cases presenting leukemic evolution demand investigation in the Brazilian population, as there's no data with regard to this theme. Thus, this study aimed to characterize and estimate the survival and leukemic evolution of patients with myelodysplastic syndromes followed up in a reference service. This is a retrospective cohort study carried out from January 2000 to December 2010. The software Epi Info 2002, version 3.5.2, was used for descriptive analysis, and for the calculations of survival probabilities the Kaplan-Meier method was employed through the software Statistic Package for Social Sciences (SPSS), version 10.0. The calculations of the association probability between the characteristics analyzed and gender were performed using the chi-square for trend, Fisher, Mann Whitney, and Log Rank tests. The significance level was 0.05. The ethical and methodological aspects of the study were approved by the Research Ethics Committee of HUOL, under the Protocol 432/10. Out of the 29 selected patients, there was a predominance of elderly people, males, with low education. They showed low cumulative probability of survival and leukemic evolution rates around 27%, with no satisfactory outcome from chemotherapy, as well as no indication of bone marrow transplantation as a possible cure. There is a need for researches with larger populations for the characterization all over the national territory.

Identification and Modification of XIAP Gene in Rat Adipose-Derived Stem Cells / 天津医药

Objective To investigate the feasibility of genetically modified X-linked inhibitor of apoptosis protein (XIAP) of rat adipose-derived mesenchymal stem cells (ADSCs) by isolating and cultivating rat ADSCs in vitro. Methods ADSCs were isolated from rat groin fat pads by collagenaseⅠdigestion under sterile condition. ADSCs were passaged and amplified with 10%FBS DMEM. The multi-differentiation potential of ADSCs was verified by cultivated with differentiation medium. XIAP expression plasmid was transfected into ADSCs. The anti-apoptotic ability of XIAP transduction was detect-ed by Western blotting assay. Results ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. Results of flow cytometry showed that there were higher expressions of CD29, CD44, CD90 and CD105 in ADSCs, which differentiated into lipocytes, chondrocytes and osteoblasts under specific conditions. There is XIAP gene modified adipose-derived mesenchy-mal stem cells Band in the corresponding molecular mass of PVDF membrane area. Conclusion ADSCs were isolated from rat subcutaneous fat pads and were easily cultivated, passaged and amplified. ADSCs can differentiate into osteoblasts, chon-drocytes and adipocytes under specific conditions, which are better resource for being used in cell therapy and tissue engi-neering.

Activation of p44/42 mitogen-activated protein kinase pathway involves in formation of hypertrophic scar and keloid / 中华医学美学美容杂志

Objective To examine the expression of p44/42 mitogen -activated protein kinase (p44/42MAPK) and Phospho-p44/42MAPK in cicatrix (hy pertropic scars and keloids) and to aim at exploring the role of activated p44/ 42 MAPK pathway in development of cicatrix. Methods In or der to analyse the differences, 10 samples of normal skin (NS), hypertropic scar s (HS) and keloids (K) were collected, and then the extracted cytoplasmic protei ns from each tissue were examined by Western blotting for p44/42 MAPK and Phosph o p44/42MAPK and immunohistochemical staining with specific antibodies was emplo yed to determine that in fibroblasts of K, HS and NS. Results There was no evident difference of p44/42MAPK in the tissues and fibroblas ts between cicatrix and NS, but Phospho-p44/42MAPK was obviously higher in cica trix than that in NS. In cicatrix, there was no evident difference of p44/42MAPK in tissues between HS and K, while in fibroblasts, Phospho-p44/42MAPK in K was much higher than in HS. Conclusion Activation of p44/42MA PK pathways involves in formation of cicatrix.

A case of myelodysplastic syndrome after oral methotrexate overdose / 대한임상병리학회지

Methotrexate is a very potent inhibitor of dihydrofolate reductase and causes bone marrow suppression and megaloblastic anemia. It is widely used in combination with other chemotherapeutic agents in lymphoproliferative disorders. A 63 year old man with ischioneuralgia developed exertional dyspnea and dizziness after he had intentionally taken methotrexate in doses of 5mg per day for 2months. Five months after discontinuation of methotrexate, his bone marrow showed the hypercellular marrow with 90% cellularity, 15% blasts and marked dysgranulopoiesis, suggestive of refractory anemia with excess blasts(RAEB). The hematopoietic cells were not enough aspirated for proper diagnosis in follow up bone marrow after three months. The bone marrow aspirates showed 13% blasts, and marked dysgranulopoiesis. The bone marrow biopsy showed hypercellular marrow with 100% cellularity, but marked fibrosis was developed. The cytogenetic study revealed normal karyotype.

Hypoplastic Acute Leukemia / 대한임상병리학회지

BACKGROUND: Hypoplastic acute leukemia is rare and most cases reported were of older age group. We reviewed our cases of hypoplastic acute leukemia and their hematologic and clinical findings. METHOD: The bone marrow biopsy slides and the reports of patients diagnosed as having acute leukemia during recent ten years were reviewed. The medical records of patients who had blast cells of greater than 30% and marrow cellularity less than or equal to 50% were reviewed. RESULTS: Of 308 patients analyzed, 17 (5.5%) fulfilled the above mentioned criteria. Ten patients were women and seven men. The median age was 44 with a range of 18-71. Chief complaints were fever, headache, general weakness and abdominal pain. Two patient presented hepatomegaly. One patient was diagnosed as granulocytic sarcoma. Ten patients were pancytopenic with median leukocyte count of 1,500/ L, hemoglobin of 8.3 g/dL, and platelet count of 27,000/ L. Circulating blast cells were 0-76%. FAB classification revealed one to be M0, three M1, seven M2, three M4, one M5, one M6 and one L1. Seven patients were not followed, and three were treated conservatively. Of seven patients receiving chemotherapy, four achieved durable complete remission. One achieved complete remission by using G-CSF. CONCLUSION: Most cases of reported hypoplastic acute leukemia were acute myelogenous leukemia of older age but our cases included leukemia of younger age and one acute lymphoblastic leukemia. Of seven patients who received chemotherapy, four achieved complete remission and one showed complete remission only by G-CSF.

Differential responses of CD34-positive acute myelogenous leukemic blasts to the costimulating effects of stem cell factor with GM-CSF and/or IL-3

Stem cell factor (SCF), a c-kit ligand, has a preferential effect on the proliferation of several classes of immature hematopoietic progenitor cells in combination with GM-CSF or IL-3. To analyze the costimulatory role of SCF in leukemic growth, we investigated the effect of SCF in the presence of GM-CSF and/or IL-3 on isolated CD34-positive (CD34+) leukemic blasts from 15 patients with acute myelogenous leukemia (AML). Cultures of CD34+ cells from normal bone marrow were used as controls. When the proliferation of CD34+ AML blasts in the presence of GM-CSF and/or IL-3 were evaluated in vitro for the effects of SCF, two patterns emerged. In one pattern, CD34+ AML blasts responded with a significant increase in DNA synthesis and/or colony formation when SCF was used with GM-CSF and/or IL-3 relative to the growth with SCF alone; This result is consistent with those CD34+ bone marrow cells from normal donors. Six patients (40%) were included in this category. The addition of SCF as a single factor resulted in colony formation in all six of these cases. In the other pattern, nine of the patients (60%) had CD34+ leukemic cells whose growth with SCF plus either GM-CSF, IL-3, or GM-CSF+IL-3, was not significantly different from the growth noted in the presence of SCF alone. Among them seven cases that did not form colonies in response to SCF alone, and one case showing autocrine, background growth were included. In the six cases in which the costimulating effects of SCF were documented, CD34+ c-kit+ blasts comprised 50.5 +/- 18.7% of the CD34+ leukemic blasts-higher than 21.8 +/- 19.4% of cases in which the costimulating effect of SCF was not documented. In the cases showing high c-kit antigen expression(> or = 40%), SCF had a costimulatory effect in 71% (5/7) of the patients. In conclusion, our data indicate that CD34+ leukemic blasts from a good proportion of patients with AML did not respond to the costimulating effects of SCF in the presence of GM-CSF adn/or IL-3, in contrast to those CD34+ bone marrow cells from normal donors. The possible use of SCF for acute leukemia must await further cytogenetic and molecular studies, which should clarify the preferential costimulating role of SCF in normal hematopoiesis.