Actividad de fosfolipasas y proteasas en aislamientos de especies de Candida colonizadoras y causantes de vulvovaginitis en mujeres gestantes / Phospholipase and proteinase activities of isolatesof colonizing Candida spp. causing vulvovaginitis in pregnant women

Introducción. Las proteasas y las fosfolipasas son factores de virulencia de Candida spp. que cumplen un papel importante en la invasión de los tejidos. Entre los factores relacionados con el huésped, se encuentran algunos asociados con las características ambientales y otros con la colonización. Objetivo. Determinar la actividad de fosfolipasas y proteasas en aislamientos de especies colonizadoras y patógenas de Candida spp., aisladas de mujeres gestantes de Cartagena de Indias. Materiales y métodos. Se determinó la actividad de fosfolipasas y proteasas en 56 aislamientos mediante degradación del sustrato y cálculo del coeficiente de actividad enzimática. Se compararon las actividades de fosfolipasas y proteasas, entre los aislamientos colonizadores y los patógenos. Resultados. La actividad de la fosfolipasa fue "muy alta" (< 0,69) en 34 aislamientos e, igualmente, la de la proteasa en 14. No hubo diferencias significativas al comparar las actividades de las fosfolipasas y de las de las proteasas, entre los aislamientos colonizadores y los patógenos. Conclusiones. La actividad de las fosfolipasas predominó como factor de virulencia en los aislamientos estudiados. No obstante, no se encontró una diferencia significativa entre los grupos de aislamientos colonizadores y los patógenos, en cuanto a las actividades de fosfolipasas y proteasas.

Introduction. Proteases and phospholipases are virulence factors of Candida spp. that play an important role in tissue invasion. Among the factors related to the host some are associated with environmental characteristics and others with Candida colonization. Objectives. To determine phospholipase and protease activities in colonizing and pathogenic strains, isolated from pregnant women in Cartagena de Indias. Materials and methods. Phospholipase and protease activity was determined in 56 isolates, evaluating substrate degradation and calculating the enzyme activity coefficient. Phospholipase and protease activities were compared between colonizing and pathogenic strains. Results. "Very high" (<0.69) phospholipase and protease activity was found in 34 and 14 isolates, respectively. There was no significant difference when comparing phospholipase and protease activities between colonizing and pathogenic isolates. Conclusions. Phospholipase activity predominated as a virulence factor in the studied strains, but no significant difference was found between colonizing and pathogenic strains for phospholipase and protease activities.

Influence of two natural cross-linkers on microtensile bond strength durability ­ An in vitro study / Influência de dois reticuladores naturais na durabilidade da resistência de união à microtração ­ Um estudo in vitro

Objective: to investigate the effect of two natural cross-linkers on microtensile bond strength (µTBS) and evaluate their influence on the durability of the resin dentin bonds. Material and Methods: the Moringa oleifera and Centella asiatica plant extracts were qualitatively tested with high-performance thin layer chromatography (HPTLC) for the presence of phenols. The phenolic content ranged from 27 to 30 gallic acid equivalents (GAE), µg/mg of dry weight. After etching, two concentrations (5% and 1%) of these two extracts were prepared and used as pretreatment liners on dentin. They were applied for a min. After restoration with resin composite, dentin resin beams were prepared. The study groups were 5% Moringa, 1% Moringa 5% Centella 1% Centella, and control (without cross-linker application). For each group, half of the samples underwent µTBS testing after 24 hours, while the remaining half were immersed in artificial saliva to assess the bond's longevity after 6 months of ageing. Statistical analysis was performed using one-way ANOVA followed by Tukey's post hoc test. Results: both 5% and 1% Moringa showed a significant difference (p<0.05) compared to the other groups at both intervals. However, after ageing, the specimens in the control and 1% Centella groups resulted in a significant decrease in µTBS. Conclusion: overall, both concentrations of Moringa (5% and 1%) were effective in stabilising the bond during both intervals.(AU)

Objetivo: investigar o efeito de dois reticuladores naturais na resistência de união (µTBS) à microtração e avaliar sua influência na durabilidade da adesão da resina à dentina. Material e Métodos: extratos das plantas Moringa oleifera e Centella asiatica foram qualitativamente testados através de cromatografia em camada fina de alta performance (HPTLC) para a presença de fenóis. O conteúdo fenólico alcançou entre 27 a 30 equivalentes de ácido gálico (GAE), µg/mg de peso seco. Após o condicionamento, duas concentrações (5% e 1%) dos extratos foram preparadas e utilizadas como forros de pré-tratamento em dentina. Eles foram aplicados por um minuto. Após a restauração com resina composta, palitos de dentina e resina foram preparados. Os grupos foram 5% Moringa, 1% Moringa, 5% Centella, 1% Centella e controle (sem aplicação de reticulador). Para cada grupo, metade das amostras foram submetidas ao teste µTBS após 24 horas, enquanto a outra metade foi imersa em saliva artificial para avaliar a longevidade adesiva após 6 meses de envelhecimento. Foi realizada análise estatística através de ANOVA 1-fator, seguido do teste post hoc de Tukey. Resultados: ambas as concentrações de 5% e 1% de Moringa demonstraram diferença significativa (p<0.05) comparadas aos outros grupos em ambos os intervalos. No entanto, após o envelhecimento, os espécimes dos geupos controle e 1% de Centella resultaram em uma redução significativa de µTBS. Conclusão: no geral, ambas as concentrações de Moringa (5% e 1%) foram efetivas em estabelecer a adesão em ambos os intervalos (AU)

Nonsense variation of OTUD5 gene in a fetus with X-linked multiple congenital anomalies- neurodevelopmental syndrome / 中华围产医学杂志

We report a fetus presented with complex cardiac malformations, pulmonary atresia with ventricular septal defect, detected by fetal echocardiography at 17 +4 weeks. The pregnancy was terminated after routine counseling and genetic tests were performed on umbilical cord of the induced fetus and peripheral blood samples of the parents. Whole-exome sequencing identified a novel maternally-inherited and likely pathogenic variation hemizygous nonsense variant, c.1651C>T (p.Gln551*) in the OTUD5 gene (NM_017602.3), which was confirmed by subsequent Sanger sequencing. The fetus was finally diagnosed as X-linked multiple congenital anomalies-neurodevelopmental syndrome.

Genes related to maintenance of autophagy and successful aging / Genes relacionados à manutenção da autofagia e envelhecimento bem-sucedido

ABSTRACT Considering aging as a phenomenon in which there is a decline in essential processes for cell survival, we investigated the autophagic and proteasome pathways in three different groups: young, older and oldest old male adults. The expression profile of autophagic pathway-related genes was carried out in peripheral blood, and the proteasome quantification was performed in plasma. No significant changes were found in plasma proteasome concentrations or in correlations between proteasome concentrations and ages. However, some autophagy- and/or apoptosis-related genes were differentially expressed. In addition, the network and enrichment analysis showed an interaction between four of the five differentially expressed genes and an association of these genes with the transcriptional process. Considering that the oldest old individuals maintained both the expression of genes linked to the autophagic machinery, and the proteasome levels, when compared with the older group, we concluded that these factors could be considered crucial for successful aging.

RESUMO Considerando o envelhecimento como um fenômeno em que há um declínio nos processos essenciais a sobrevivência celular, investigamos as vias autofágica e proteassômica em três grupos: jovens, idosos e longevos. O perfil de expressão dos genes relacionados à via autofágica foi analisado em sangue periférico, e a quantificação do proteassoma realizada em plasma. Não foram encontradas alterações significativas nas concentrações plasmáticas de proteassoma ou na correlação entre as concentrações de proteassoma e as idades. No entanto, alguns genes relacionados a autofagia e / ou apoptose foram expressos diferencialmente. Além disso, as análises de rede e de enriquecimento mostraram uma interação entre quatro dos cinco genes diferencialmente expressos e a associação desses ao processo transcricional. Considerando que os indivíduos longevos mantiveram tanto a expressão de genes ligados à maquinaria autofágica, quanto os níveis de proteassoma quando comparados aos idosos, concluímos que esses fatores poderiam ser considerados cruciais para o envelhecimento bem-sucedido.

Study on expression of ASC and Caspase-1 in peripheral blood of patients with primary biliary cirrhosis / 重庆医学

Objective To explore the influence of apoptosis-associated speck-like protein containing a caspase recruitment do-main(ASC)and Caspase-1 on the pathogenesis of primary biliary cirrhosis(PBC).Methods The real-time PCR,Western blot,py-rophosphate sequencing and ELISA were adopted to respectively detect the relative expressions of mRNA and protein of peripheral blood mononuclear cells(PBMS)Caspase-1 and ASC as well as the methylation status of ASC promoter region and plasma Caspase-1 and IL-18 expression levels in 30 cases of PBC and healthy controls.Results The mRNA and protein expressions of PBMC Caspase-1 and ASC in the PBC group were significantly higher than those in the control group(P<0.05).The methylation rate of ASC promoter Island1(ISI)was significantly lower than that of the control group(P<0.05),which of Island 2(IS2)was smaller than the background value and had no methylation occurrence.The levels of plasma Caspase-1 and IL-18 in the PBC group were sig-nificantly higher than those in the control group(P<0.05).The ASC mRNA in the PBC group was significantly correlated with the Caspase-1 mRNA expression(P<0.05);the methylation rates at loci 1,2,4,5 of ASC promoter region CpG island were nega-tively correlated with ASC mRNA expression(P< 0.05),and which at loci 3,6 had no correlation with their expressions(P>0.05);plasma Caspase-1 and IL-18 levels showed the obviously positive correlation.Conclusion ASC and Caspase-1 are involved in the immune inflammatory response in PBC.

The role of UCH-L3 and LC3Ⅱin the diagnosis of acute myeloid leukemia / 中华检验医学杂志

Objective Todetect the mechanism of acute myeloid leukemia ( AML ) and the relationship between autophagy andubiquitin c-terminal hydrolases L3( UCH-L3) for providing new targets and guiding clinical management in AML .Methods 84 cases of AML patients and 25 controls were chosen from the First Affiliated Hospital of Dalian Medical University from 2014 to 2015,including 47 males and 37 females.The AML patients were divided into 3 groups :initialdiagnosis group [40 cases including 24 males and 16 females,with an median age of 54 (23 -85)];complete remission group [30 cases including 14 males and 16 females,with an median age of 45 (22-74)]and refractory group[14 cases including 9 males and 5 females,with an median age of 50 (18-80)].Among those patients, there were 3 cases of M1, 42 cases of M2, 18 cases of M3, 3 cases of M4 and 18 cases of M5 subtypes.The expression of UCH-L3 and LC3II were detected by Real Time PCR and Western blot after the HL-60 cells were treated by TCID.The expression of UCH-L3 and LC3IIin the PBMC of AML patients and controls were detected with Real Time PCR.The expression levels of UCH-L3 and LC3II in different types of AML were analyzed .The relationship between the expression of LC 3Ⅱand clinical features , clinical stages , laboratory results and therapeutic effects of patients were investigated .It further detected the expression of UCH-L3 and LC3IImRNA in post-induction status.t test were used for measurement data , χ2 test were performed for rate comparison; rank correlation test were performed for correlation analysis .Non-parametric test were performed for non-normal distribution data.Results Both the UCH-L3 and LC3 II were down-regulated after TCID treatment in HL-60 cellsat gene and protein levels (t=-29.435, t=-8.105,P<0.05).The expression of UCH-L3 in initial diagnosis group was lower than control group ( Z=-3.87,P<0.05),butit was higher in remission group than initial diagnosis group and refractory group ( Z=-6.70 , Z=-4.09, P<0.05 ) .The expression level of LC3Ⅱin diagnosis group was significantly higher than control group , remission group and refractory group(Z=-6.96,Z=-5.32, Z=-3.52,P<0.05).Cytogenetic abnormalities in patients with high expression of LC3II were more common(χ2 =6.510,P<0.05).The relative expression of UCH-L3 and LC3ⅡmRNA in the same patient before and after treatment were significantly different ( P<0.05 ) . During the remission, the expression of UCH-L3 was up-regulated compared with that before primary treatment, while the expression of LC3II was down-regulated.There was no statistically significant in the relative expression of UCH-L3 and LC3Ⅱamong the FAB type.Conclusion The UCH-L3 and autophagy are associated with pathogenesis of AML .

Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy / 国际脑血管病杂志

Cerebral small vessel disease is a group of heterogeneous diseases with stroke and cognitive impairment as the main clinical features.It can be divided into sporadic type and hereditary type.Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) belongs to hereditary cerebral small vessel disease.It has been reported in China,Japan,Spain,Greece,and other countries.The diagnosis mainly depends on characteristic clinical symptoms,imaging features,and genetic testing.CARASIL manifests as diffuse white matter abnormal signal and subcortical multiple infarcts on MRI,and it caused by HTRA1 gene mutation.

Corin-serine protease and clinical applications in cardiovascular disease / 中华检验医学杂志

Corin is a newly identified membrance protease which responsible for converting pro-B-type natriuretic peptide to biologically active B-type natriuretic peptide.B-type natriuretic peptide play an important role in regulating blood pressure and cardiac function.It is a biological marker of heart failure.So Corin plays a role in reducing blood volume, blood pressure, regulating body fluid balance, and improving cardiac function.Corin may be used as a biomarker for heart failure or other cardiovascular disease.Recent studies have found that Corin is associated with the occurrence and development of cardiovascular diseases, such as heart failure and acute coronary syndrome.The article mainly expound the biological characteristics of Corin, testing methods and its clinical application in cardiovascular disease status.

Effect of mild hypothermia on expression of small ubiquitin-like modifier-specific proteases 3 in brain tissues during cerebral ischemia-reperfusion in mice / 中华麻醉学杂志

Objective To investigate the effect of mild hypothermia on the expression of small ubiquitin-like modifier-specific proteases 3 (SENP3) in the brain tissues during cerebral ischemia-reperfusion (I/R) in mice.Methods Ninety-six male C57/BL6 mice,aged 10-12 weeks,weighing 22-30 g,were randomly divided into 3 groups (n =32 each) using a random number table:sham operation group (group S),group I/R,and mild hypothermia group (group H).Cerebral I/R was produced by occlusion of bilateral common carotid arteries for 15 min followed by reperfusion.The surface cooling was started immediately after reperfusion,and the rectal temperature was maintained at 32-34 ℃ for 3 h.At 6,12,24 and 72 h of reperfusion,8 mice were selected from each group and sacrificed.The hippocampi were removed for examination of the pathological changes (with light microscope) and for determination of cell apoptosis (using TUNEL) and expression of SENP3 (by Western blot).The apoptosis rate was calculated.Results Compared with group S,the apoptosis rate in hippocampal CA1 region was significantly increased,and the expression of SENP3 was significantly up-regulated at each time point of reperfusion in group I/R (P＜0.05).Compared with group I/R,the apoptosis rate in hippocampal CA1 region was significantly decreased,and the expression of SENP3 was significantly down-regulated at each time point of reperfusion (P＜0.05),and the pathological changes were significantly reduced in group H.Conclusion The mechanism by which mild hypothermia reduces cerebral I/R injury is associated with inhibition of SENP3 expression in the brain tissues of mice.

Role Of MMP-2 and MMP-9 in Resistance to Drug Therapy in Patients with Resistant Hypertension / Papel da MMP-2 e MMP-9 na Resistência à Terapia Medicamentosa em Pacientes com Hipertensão Arterial Resistente

AbstractBackground:Despite the increased evidence of the important role of matrix metalloproteinases (MMP-9 and MMP‑2) in the pathophysiology of hypertension, the profile of these molecules in resistant hypertension (RHTN) remains unknown.Objectives:To compare the plasma levels of MMP-9 and MMP-2 and of their tissue inhibitors (TIMP-1 and TIMP-2, respectively), as well as their MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios, between patients with controlled RHTN (CRHTN, n=41) and uncontrolled RHTN (UCRHTN, n=35). In addition, the association of those parameters with clinical characteristics, office blood pressure (BP) and arterial stiffness (determined by pulse wave velocity) was evaluate in those subgroups.Methods:This study included 76 individuals diagnosed with RHTN and submitted to physical examination, electrocardiogram, and laboratory tests to assess biochemical parameters.Results:Similar values of MMP-9, MMP-2, TIMP-1, TIMP-2, and MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios were found in the UCRHTN and CRHTN subgroups (P>0.05). A significant correlation was found between diastolic BP (DBP) and MMP-9/TIMP-1 ratio (r=0.37; P=0.02) and DPB and MMP-2 (r=-0.40; P=0.02) in the UCRHTN subgroup. On the other hand, no correlation was observed in the CRHTN subgroup. Logistic regression models demonstrated that MMP-9, MMP-2, TIMP-1, TIMP-2 and their ratios were not associated with the lack of BP control.Conclusion:These findings suggest that neither MMP-2 nor MMP-9 affect BP control in RHTN subjects.

ResumoFundamento:A despeito da crescente evidência do importante papel das metaloproteinases da matriz extracelular (MMP-9 e MMP-2) na fisiopatologia da hipertensão, o perfil dessas moléculas na hipertensão arterial resistente (HAR) permanece desconhecido.Objetivo:Comparar os níveis plasmáticos de MMP-9 e MMP-2 e seus inibidores teciduais (TIMP-1 e TIMP-2, respectivamente), assim como as suas razões MMP-9/TIMP-1 e MMP-2/TIMP-2, entre pacientes com HAR controlada (HARC, n = 41) e HAR não controlada (HARNC, n = 35). Além disso, a associação desses parâmetros com as características clínicas, pressão arterial (PA) de consultório e rigidez arterial (determinada pela velocidade da onda de pulso) foi avaliada nesses subgrupos.Métodos:Este estudo incluiu 76 indivíduos com HAR submetidos a exame físico, eletrocardiografia e exames laboratoriais para a avaliação de parâmetros bioquímicos.Resultados:Valores semelhantes de MMP-9, MMP-2, TIMP-1, TIMP-2, e razões MMP-9/TIMP-1 e MMP-2/TIMP-2 foram encontrados nos subgrupos HARNC e HARC (p > 0,05). Observou-se uma correlação significativa entre PA diastólica (PAD) e razão MMP-9/TIMP-1 (r = 0,37; p = 0,02) e PAD e MMP-2 (r = -0,40; p = 0,02) no subgrupo HARNC. Por outro lado, não se observou correlação no subgrupo HARC. Os modelos de regressão logística demonstraram que MMP-9, MMP-2, TIMP-1, TIMP-2 e suas razões não se associaram com a falta de controle da PA.Conclusão:Esses achados sugerem que MMP-2 e MMP-9 não afetem o controle da PA em indivíduos com HAR.

Expression of PACE4 in non-small cell lung cancer and analysis of its clinicopathologic characteristics / 中国医师杂志

Objective To investigate the expression of paired amino acid cleaving enzyme 4 (PACE4) protein in non-small-cell lung cancer (NSCLC), analyze the correlation of its clinicopathologic characteristics, and investigate its significance in the process of occurrence, development, and metastasis of NSCLC.Methods Between January 2006 and May 2009, the First Affiliated Hospital of Guangzhou Medical College, 172 patients with NSCLC and 15 patients with non-neoplastic tissues were chosen.Immunohistochemistry was used to detect the expression of PACE4, and clinicopathologic characteristics of NSCLC were analyzed.Results (1)The positive expression rate of PACE4 protein in NSCLC was significantly higher than that in non-tumor tissues (P ＜ 0.05).PACE4 expression was observed in 111 of 172 (64.5％) NSCLC.PACE4 had cytoplasmic expression.(2)Clinicopathologically, PACE4 expression was significantly associated with lymph node metastasis (N stage) (P =0.007), and clinical stage (P =0.024).Conclusions High expression of PACE4 indicates that PACE4 may be involved in the processes of occurrence,development, and metastasis of NSCLC.

Liensinine promotes apoptosis of bladder cancer 5637 cells and enhances Caspase-7 expression and activation / 天津医药

Objective To study the effects of Liensinine on apoptosis of 5637 cells, and its mechanism thereof. Meth?ods CCK-8 method and the colony formation test were used to detect cell viabilities, and then inhibition rates were calcu?lated. Flow cytometry was used to detect the effects of Liensinine on apoptosis of 5637 cells. Western blot assay was used to detect Caspase-7 protein expression. Results CCK-8 assay and colony formation test indicated that Liensinine inhibited the cell proliferation significantly. Results of flow cytometry indicated that Liensinine induced early apoptosis of 5637 cells. Western blot assay showed that Liensinine improved the expression of Caspase-7 and enhanced the activation of Caspase-7 in 5637 cells. Conclusion Liensinine could inhibit the proliferation of 5637 cells, induce early apoptosis, which may be re?lated with the enhanced expression of Caspase-7 and its activation.

The relationship between PCSK9 gene I474V polymorphism and ischemic stroke of Uygur and Han ethnic groups in Xinjiang Uygur Autonomous Region / 中华神经科杂志

Objective To investigate the frequency distribution of proprotein convertase subtilisin/kexin 9 (PCSK9) gene I474V polymorphisms and their relationship with patients with ischemic stroke (IS)of Uygur and Han ethnic groups in Xinjiang Uygur Autonomous Region.Methods The I474V polymorphism was identified by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) in 407 patients with IS(including 219 Hans and 188 Uygurs)and 425 health controls (including 255 Hans and 170 Uygurs),and some specimens were sequenced.Results (1) Between IS group and control group,the genotypes Ⅱ and Ⅳ had no statistically significant differences in the levels of triglycerides (TG),high-density lipoprotein cholesterol (HDL-C) ; Total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C) levels had statistically significant differences; LDL-C levels had also statistically significant differences.Between IS and control groups,TC,LDL,HDL-C levels of genotype Ⅱ showed statistically significant difference.In the IS group,TC,LDL-C levels of Ⅳ genotype were significantly higher than the control group,the difference being statistically significant.(2) There was statistically significant difference in the genotype distribution between IS and control groups (9.5％ (77/814) vs 4.5％ (38/850),x2 =16.09,P =0.000).And the distribution of allele frequency was statistically different (18.9％ (77/407) vs 8.9％ (38/425),x2 =17.38,P =0.000).(3) The differences of I474V loci Ⅳ genotype frequency distribution in Xinjiang Uygurs and Hans were statistically significant (27.7％ (52/188) vs 11.4％ (25/219),x2 =17.40,P =0.000; 12.9％ (22/170) vs 6.3％ (16/255),x2 =5.57,P =0.018) ; So did the Ⅴ allele frenquency distribution (13.8％ (52/376) vs 5.7％ (25/438),x2 =15.58,P =0.000; 6.5％ (22/340) vs 3.1％ (16/510),x2 =10.44,P =0.001).(4) There was statistically significant difference in the genotype distribution and allele frenquency distribution between IS group and control group in the Xinjiang Uygurs (27.7％ (52/188) vs 12.9％ (22/170),x2 =11.79,P =0.001 ; 13.8％ (52/376) vs 6.5％ (22/340),x2 =10.44,P =0.001) ; But there was no statistically significant difference in the Hans.Conclusions Ⅱ and Ⅳ genotypes are dominant in the I474V polymorphism loci of PCSK9 gene.The genotype of PCSK9 gene I474V polymorphism is correlated with increasing serum levels of TC and LDL-C.I474V polymorphism is associated with cerebral IS course in Xinjiang region.There is statistically significant difference in the genotype I474V distribution between Uygur and Han groups.I474V polymorphism has a relationship with the occurrence of IS in Xinjiang Uygurs.Ⅳ may be a susceptible genotype and Ⅴ may be a genetic susceptible allele of the Xinjiang Uygurs.

Proteinase activity in latex of three plants of the family Euphorbiaceae

In the family of Euphorbiaceae, the genera Euphorbia and Sapium are known to contain essentially latex-bearing species. In the present study, the latex of Euphorbia selloi (Klotzsch & Garcke) Boiss., Euphorbia papillosa A.St.-Hil., and Sapium glandulosum (L.) Morong, plants native from Brazil, were examined concerning proteolytic activity. All studied species have proteins with significant proteolytic activity and E. papillosa has the greatest specific activity. Aiming to verify the type of protease present, an assay with different inhibitors was performed. In the three tested plants, the proteolytic activity was significantly inhibited by a serine protease inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF). Using techniques of electrophoresis with polyacrylamide gels (SDS-PAGE), the subunits of proteins were separated according to their molecular masses, and the protein activity was visually detected by zymography.

Dentro da família Euphorbiaceae, os gêneros Euphorbia e Sapium são conhecidos por incluírem basicamente espécies produtoras de látex. No presente estudo, o látex das plantas Euphorbia selloi (Klotzsch & Garcke) Boiss., Euphorbia papillosa A.St.-Hil. e Sapium glandulosum (L.) Morong, espécies nativas do Brasil, foi analisado em relação à atividade proteolítica. Todas as amostras analisadas possuem proteínas com significativa atividade, sendo que o látex da espécie E. papillosa apresenta a maior atividade específica. Com o objetivo de analisar quais os tipos de proteases responsáveis pela atividade proteolítica, realizaram-se ensaios com diferentes inibidores. Nas três plantas testadas a atividade foi inibida significativamente pelo cloridrato de 4-(fluoreto de 2-aminoetilbenzenossulfonil) (AEBSF), um inibidor de serino-proteases. Utilizando técnicas de eletroforese em gel de poliacrilamida (SDS-PAGE), as subunidades das proteínas foram separadas de acordo com sua massa molecular e, através da zimografia, a atividade proteolítica pode ser detectada visualmente.

A Novel PHEX Gene Mutation in a Patient with Sporadic Hypophosphatemic Rickets

Phosphate regulating gene with homologies to endopeptidases on the X-chromosome (PHEX) is a common cause of X-linked hypophosphatemic (XLH) rickets. Diverse PHEX gene mutations have been reported; however, gene mutations in sporadic rickets are less common than in XLH rickets. Herein, we describe a 50-year-old female patient with sporadic hypophosphatemic rickets harboring a novel splicing-site mutation in the PHEX gene (c.663+1G>A) at the exon 5-intron 5 boundary. The patient had recently suffered from right thigh pain and an aggravated waddling gait. She also presented with very short stature, generalized bone pain, and muscle weakness. Despite low serum phosphate levels, her phosphate reabsorption rate was lower than normal. Additionally, her 1,25-dihydroxyvitamin D3 concentration was lower than normal, although FGF23 level was normal. After treatment with alfacalcidol and elemental phosphate, her rachitic symptoms subsided, and callus formation was observed in the fracture site on the right femur.

Effects and mechanisms of trophoblast cells autophagy in preeclampsia / 中华妇产科杂志

Objective To investigate the occurrence law of autophagy in trophoblast cells from preeclampsia and its underlying mechanisms.Methods Twenty cases of placenta tissues were collected from women suffered from preeclampsia and normal pregnant women respectively.Autophagosome of trophoblast cells were observed by transmission electron microscope.The expressions of LC3-Ⅱ/Ⅰ and Atg4B in placenta tissues were detected by western blot and real-time PCR.Results Compared with the control group,typical autophagosomes of trophoblast cells were observed by transmission electron microscope.The ratio of LC3-Ⅱ / Ⅰ in placenta of PE patients was increased (1.43 ± 0.23) compared with control group (0.59 ±0.12),and the expression of Atg4B was up-regulated in both mRNA [(1.73 ±0.16) folds] and protein levels (0.71 ± 0.13) compared with control group (P ＜ 0.05).Conclusions Autophagy was significantly up-regulated in trophoblast cells from patients suffered from preeclampsia.Thus,all the data suggest that autophagy might be involved in the generation of preeclampsia.

Comparative study of two proteases for mass spectrometric method for the determination HbA1c / 中华检验医学杂志

Objective To explore the difference of two kinds of proteolytic enzymes (GLU-C and Lys-C trypsin),hemoglobin A1c were measured in mass spectrometry.Methods Based on the IFCC recommended hemoglobin A1c reference measurement method by mass spectrometry,the blood samples were preparied (the number of these blood samples:201201,201202,201203).Hemoglobin in the samples were enzymed respectively by two kinds of proteolytic enzymes (Glu-C and Lys-C),before the mass spectra which be used solid phase extraction method,under the optimal experimental conditions,hemoglobin A1c in the sample were measured by the liquid chromatography tandem mass spectrometry and screened of variant hemoglobin with sequest software.All results by mass spectrometry are required to use the Sequest software to search RAW file in the human IPI Database,retrieve four common hemoglobin variant published results and compare hemoglobin variant sites,using statistical software SPSS13.0 and Excel 2003 on the results in the correlation analysis,compare between groups by the independent sample t test.Results In the detection of hemoglobin A1c,respectively Glu-C and Lys-C blood which the samples were enzymed,results of hemoglobin A1c by Lys-C were (34.70 ± 2.80),(51.76 ± 1.60),(73.39 ± 1.11) mmol/mol,results of hemoglobin A1c by Glu-C were(33.12 ± 1.48),(54.54 ± 2.50),(75.40 ± 3.60) mmol/mol,the difference between groups was not statistically significant,the results of two methods of drawing curve,all R2 ＞ 0.995.Searching the human database with sequest software,four common hemoglobin variant were not be finded.Conclusions In the detection of hemoglobin A1c by the mass spectrometry,the application of the above two kinds of proteolytic enzyme in the blood samples,that can be more consistent results,test results of two methods have a good correlation.

Expression and significance of matriptase in ovarian cancer cells with diverse metastatic potential / 中华妇产科杂志

Objective To study the expression and significance of matriptase in different metastatic potential of human ovarian cancer cells.Methods High-metastatic human ovarian cancer cell HO8910PM and ovarian cancer cell HO8910 were collected.The ability of metastatic of the former was stronger than that of the latter.Compared the ability of invasion and migration in HO8910PM and HO8910 by scratch assay and by millicell chamber artificial reconstituted basement membrane invasion assay.Detected the matriptase mRNA and protein expression levels in HO8910PM and HO8910 through reverse transcription(RT)-PCR and immunocytochemistry methods.Results The 24 hours' migration distance(347 ± 8) μm of HO8910PM cells were significantly higher than that in HO8910 group (154 ± 10) μm (P ＜ 0.01) ;The number of HO8910PM cells that penetrated the matrigel after 24 hours' incubation were significantly higher than that in HO8910 group (90.7 ±2.1 vs 63.3 ± 1.5,P ＜0.01).The expression of matriptase mRNA in HO8910PM cells was higher than that in HO8910 group (0.72 ± 0.03 vs 0.38 ± 0.04,P ＜ 0.01).The migration was positively correlated with the matriptase mRNA expression levels (r =0.992,P ＜ 0.01); and the invasiveness was also positively correlated with the matriptase mRNA expression levels (r =0.973,P ＜0.01).As far protein level,the expression of matriptase protein in HO8910PM cells was higher than that in HO8910 group (15.6 ±0.8 vs 7.6 ± 1.3,P ＜0.01).The migration was positively correlated with matriptase protein expression levels (r =0.971,P ＜ 0.01) ;And the invasiveness was also positively correlated with the matriptase protein expression levels (r =0.958,P ＜ 0.01).Conclusions The relationship between the expression levels of matriptase and the metastatic of ovarian cancer cells may be correlative.The function of matriptase in ovarian cancer cells metastatic machanism still need to be confirmed.

The correlation between aggrecan degradation and the progression in relapsing polychondritis disease / 中华检验医学杂志

Objective To explore the significance in judging the different clinical stages of relapsing polychondritis (RP) patients through examining the changes of aggrecanase and metabolic fragments of aggrecan.MethodsIn comparison with the control group (20 cases),40 patients were divided into the stable stage group (22 cases) and the active stage group (18 cases).The aggrecanase-generated neoeptitopes in cartilage matrix were analysed by immunohistochemistry and Western blot(WB) respectively.The mRNA and protein levels of aggrecanase-1,2 expressed in cartilage cells were measured by real-time reverse transcriptional polymerase chain reaction(RT-PCR) and WB respectively.The difference of these results among these three groups was analyzed accordingly.ResultsThe expression of aggrecanase-1,2 in mRNA level was measured by real-time RT-PCR.The values of aggrecanase-1,2 mRNA 2 -ΔΔC1 were 1.00 ± 0.26 and 1.00 ± 0.27 in control group,1.47 ± 0.11 and 1.57 ± 0.13 in stable stage group,2.09 ±0.12 and 2.09 ± 0.19 in active stage group respectively.By one-way ANOVA analysis,the difference between every two groups was statistically significant (F was 299.113 and 459.013,P ＜ 0.01 ).In comparison with control group,aggrecanase-1,2 increased significantly in both stable and active stage group (P ＜ 0.01 ) and aggrecanase-1,2 increased more significantly in active stage group than in stable stage group (P ＜ 0.01 ).The results from WB analysis indicated that aggrecanase-1,2 could not be detected in control group,and they were detectable in stable stage group and increased in active stage group at the relative molecular of 68 000 Da or 73 000 Da respectively.The aggrecanase-generated neoeptitopes were analyzed by WB as well.The results indicated that NITEGE and ARGSV could be detected in stable stage group and increased in active stage group at the relative molecular of 70 000 Da or 48 000 Da respectively,but there were no signals in control group.Similar with the previous WB results,no signals of NITEGE or ARGSV eptitopes were detected in normal cartilage matrix ( no red staining) by use of immunohistochemical staining.However,in stable stage group and active stage group,these eptitopes were apparently detected (obviously red staining).ConclusionWith the progression of the RP,the activity of the aggrecanase is enhanced,and the degradation of the aggrecan is increased,associated with the severity of the disease.

Cerebral autosomal recessive arteriopathy with subcortical infarets and leukoencephalopathy / 国际脑血管病杂志

Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is an autosomal recessive hereditary cerebral artery disease in the adolescence,and its main clinical manifestations are dementia,stroke,low back pain and alopecia.At present,most cases are from Japan.3-6 exon point mutations in the HTRA1 gene are associated with the onset of CARASIL.Brain histopathological examination showed a small arterial intimal thickening medial smooth muscle cell loss and hyalinization.Brain MRI showed a diffuse white matter abnormal signal and multiple subcortical infarcts.The diagnosis mainly depends on the characteristic clinical symptoms,imaging characteristics and genetic testing.It should be differentiated from cerebral autosomal dominant arteriopathy with sulcortical infarcts and leucoencephalopathy.