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Abstract We report here microemulsions (MEs) for topical delivery of protoporphyrin IX (PpIX) for Photodynamic Therapy (PDT) of skin cancers. Selected MEs consisting of Oil/Water (O/W) bicontinuous (BC) and Water/Oil (W/O) preparations were characterized as to pH, nanometric size, zeta potential, drug content, and viscosity. Sustained in vitro PpIX release was achieved from MEs 2A (O/W), 10B (BC) and 16B (W/O) through an artificial membrane for up to 24 h, characterizing MEs as drug delivery systems. None of these MEs showed permeation through the skin, demonstrating the required topical effect. After 4 h, in vitro retention of PpIX in the stratum corneum (SC) was higher from both ME 10B and control (PpIX at 60 µg/mL in PEG 300). However, in the Epidermis + Dermis ([Ep + D]), retention from ME 10B and ME 16B was ~40 times higher compared to control. Confocal Laser Scanning Microscopy (CLSM) showed higher fluorescence intensity in the SC for both control and ME 10B, whereas ME 10B fluorescence was higher in [Ep+D]. The results indicate that ME 10B is suitable for PpIX encapsulation, showing good characteristics and a localized effect for a potential delivery system for PDT-associated treatments of skin cancers.
Subject(s)
Photochemotherapy/adverse effects , Protoporphyrins/agonists , Skin/injuries , Skin Neoplasms/pathology , In Vitro Techniques/instrumentation , Pharmaceutical Preparations/administration & dosage , Microscopy, Confocal/methods , Dermis/abnormalitiesABSTRACT
The study of the interaction of synthetic protoporphyrin IX (PpIXs) and protoporphyrin IX extracted from Harderian glands of ssp Rattus novergicus albinus rats (PpIXe) with bovine serum albumin (BSA) was conducted in water at pH 7.3 and pH 4.5 by optical absorption and fluorescence spectroscopies. PpIXs is present as H- and J-aggregates in equilibrium with themselves and with monomers. The PpIXs charge is 2− at pH 7.3 and 1− at pH 4.5. This increases its aggregation at pH 4.5 and shifts the equilibrium in favor of J-aggregates. In spite of electrostatic attraction at pH 4.5, where BSA is positive, the binding constant (Kb) of PpIXs to BSA is 20% less than that at pH 7.3, where BSA is negative. This occurs because higher aggregation of PpIXs at pH 4.5 reduces the observed Kb value. At both pHs, water-soluble PpIXe exists in the monomeric form with the charge of 1− and its Kb exceeds that of PpIXs. At pH 4.5, its Kb is 12 times higher than that at pH 7.3 due to electrostatic attraction between the positively charged BSA and the negatively charged PpIXe. The higher probability of PpIXe binding to BSA makes PpIXe more promising as a fluorescence probe for fluorescence diagnostics and as a photosensitizer for photodynamic therapy. The existence of PpIXe in the monomeric form can explain its faster cell internalization. Aggregation reduces quantum yields and lifetimes of the PpIXs excited states, which explains higher phototoxicity of PpIXe toward malignant cells compared with PpIXs.
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ABSTRACT In this study, children with sickle cell anemia were evaluated for iron deficiency. Serum ferritin and free erythrocyte protoporphyrin free erythrocyte protoporphyrin (FEP) levels, mean corpuscular volume mean corpuscular volume (MCV) and mean corpuscular hemoglobin mean corpuscular hemoglobin (MCH) were used in determining their iron status. The study was done at Pediatric Hematology Outpatient Clinic of the Obafemi Awolowo University Teaching Hospitals' Complex, Ile-Ife. Forty-eight HbSS subjects in steady state and 48 apparently well age and sex matched HbAA controls were evaluated. Serum ferritin less than 25 ng/dL FEP greater than cut off for age, mean corpuscular volume MCV and mean corpuscular hemoglobin MCH less than cut off for age were regarded as indicating iron deficiency. Serum ferritin values ranged from 34.2 to 3282.9 µg/L, with a mean of 381.2 (1.0), median 180 µg/L; which was significantly higher than the controls (p = 0.000). FEP was lower in the subjects but none was iron deficient compared with the controls. The mean corpuscular hemoglobin MCH of subjects was significantly lower than the controls. Subjects had lower mean corpuscular volume MCV compared with controls. Iron deficiency was not detected in any of the subjects with sickle cell anemia in comparison to a prevalence of 43.75% in the controls. Iron deficiency anemia (IDA) was found in 16.7% of the controls, using the WHO cut off for anemia which is hemoglobin concentration of <11 g/dl. While a high prevalence of iron deficiency was noted in the control group, patients with sickle cell anemia were largely iron sufficient, despite their anemia. Iron supplementation remains unnecessary as part of routine management of children with sickle cell anemia in our practice.
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Protoporphyrins , Anemia, Iron-Deficiency , Ferritins , Anemia, Sickle CellABSTRACT
Objective To explore the roles of heme oxygenase-1 (HO-1 ) and protoporphyrin zinc IX (ZnPPIX ) , its inhibitor , in cisplatin chemotherapy for gastric cancer so as to provide potential targets for chemosensitivity in gastric cancer .Methods Gastric cancer cell line SGC7901 was used in vitro .MTT assay was carried out to determine the effects of ZnPPIX and CDDP on the proliferation in gastric cancer cells .The expression of HO-1 in gastric cancer cells was measured by Real-time PCR and Western blot ,respectively .The gastric cancer xenografts in nude mice were used to study the effects of ZnPPIX and CDDP in gastric cancer on tumor formation in vivo .Results The proliferation of cancer cells ,interfered by CDDP in combination with ZnPPIX ,could be significantly inhibited (P<0 .05) .Moreover ,CDDP could increase the expression of HO-1 in gastric cancer cells , which was reversed by ZnPPIX (P<0 .05) .The animal experiment showed that CDDP could inhibit gastric cancer growth in nude mice and reduce tumor volume and weight . Conclusion ZnPPIX could enhance the chemosensitivity of CDDP in gastric cancer ,which may be a potential sensitizer of cisplatin-based chemotherapy in gastric cancer .
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Objective To investigate the value of Cells Free Ferrous Protoporphyrin (FH) detection in cervical cancer screening.Methods Retrospective analysis of December 2013 to 2014 year in December to accept the clinical data of 368 cases of female cervical cancer screening in Ningxia People's Hospital of Zhongwei Prefecture, divided into normal group (n=210), cervical precancerous lesions group (n=130) and cervical carcinoma group (n=28).The liquid based thin-layer cell detection (TCT) and FH detection were carried out, and the detection effects of the 2 detection methods were observed and compared.Results The statistics and comparison, in the detection of normal cervix, the false positive rate of FH detection was 19.05%, significantly higher than the detection of TCT 4.76% (P< 0.05);in the detection of cervical precancerous lesions, FH positive detection rate was 86.15%, lower than the detection of TCT 86.92%, and with the degree of cervical lesions continuously improve the positive rate of the 2 detection methods are showing a rising situation;in the detection of cervical cancer, the positive rate of FH detection was 96.43%, higher than the detection of TCT 92.86%, but the difference was not significant.Conclusion With ThinPrep cytology test (TCT), FH in the detection of relative detection effect of normal cervix of the poor, but in precancerous lesions and cervical cancer detection and detection of TCT are basically the same, therefore, FH still has a certain application value in screening of cervical cancer.
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Objective:To explore the effects of protoporphyrin Ⅸ (PpⅨ)-mediated photodynamic therapy (PDT) on induction of apoptosis and death in colon cancer cell and the underlying mechanisms.Methods:The cell killing effect of PDT on HCT116 cell was determined by cell counting kit (CCK).The cells were divided into a control group,a single light group,a single PpⅨ group,and a PDT group.Hoechst 33342 and flow cytometry was used to assess the cell apoptosis.Western blot was employed to analyze the expressions ofbd-2,bax,and caspase-3.Reactive oxygen species (ROS) was detected by flow cytometry.Results:The viability of HCT116 cell was decreased gradually with the increase of irradiation dose (P<0.05).Compared to the other 3 groups,ROS production,the number of apoptotic cells and the protein expressions ofbax and caspase-3 in PDT group increased,while bcl-2 expression was decreased (P<0.05).Conclusion:PpⅨ-mediated PDT can enhance the apoptosis in HCT116 cell,which may be related to mitochondrial apoptosis pathway.
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OBJECTIVE@#To investigate the role of human host heme-oxygenase-1 (HO-1) in pathogenesis of cerebral malaria in the in vitro model.@*METHODS@#The effect of human host HO-1 [human brain microvascular endothelial cell (HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells (iRBC) through measurement of the enzymatic products iron and bilirubin.@*RESULTS@#Following exposure to the HO-1 inducer CoPPIX at all concentrations, the HBMEC cells apoptosis occurred, which could be prominently observed at 15 μM of 3 h exposure. In contrast, there was no significant change in the morphology in the non-exposed iRBC at all concentrations and exposure time. This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin, of which the highest levels (106.03 and 1753.54% of baseline level, respectively) were observed at 15 μM vs. 20 μM at 3 h vs. 24 h exposure. For the effect of the HO-1 inhibitor ZnPPIX, HBMEC cell morphology was mostly unchanged, but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h (37.17% of baseline level). The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed (highest effect at 10 μM and 3 h exposure).@*CONCLUSIONS@#Results provide at least in part, insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.
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Objective To investigate the role of human host heme-oxygenase-1 (HO-1) in pathogenesis of cerebral malaria in the in vitro model. Methods The effect of human host HO-1 [human brain microvascular endothelial cell (HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells (iRBC) through measurement of the enzymatic products iron and bilirubin. Results Following exposure to the HO-1 inducer CoPPIX at all concentrations, the HBMEC cells apoptosis occurred, which could be prominently observed at 15 μM of 3 h exposure. In contrast, there was no significant change in the morphology in the non-exposed iRBC at all concentrations and exposure time. This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin, of which the highest levels (106.03 and 1753.54% of baseline level, respectively) were observed at 15 μM vs. 20 μM at 3 h vs. 24 h exposure. For the effect of the HO-1 inhibitor ZnPPIX, HBMEC cell morphology was mostly unchanged, but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h (37.17% of baseline level). The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed (highest effect at 10 μM and 3 h exposure). Conclusions Results provide at least in part, insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.
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As a key regulator of melanogenesis, p53 controls microphthalmia-associated transcription factor (MITF) and tyrosinase expression. The anti-oxidant enzyme heme oxygenase-1 (HO-1) is induced by various forms of cellular stress and diverse oxidative stimuli. However, few studies have examined the role of HO-1 in melanogenesis. Therefore, the aim of this study was to determine the role of HO-1 in melanogenesis and the mechanism underlying this relationship. Cultures of normal human melanocytes were treated with the HO-1 inducer cobalt protoporphyrin (CoPP) or the HO-1 inhibitor zinc protoporphyrin (ZnPP). We then measured the melanin content of the cells. Additional analyses consisted of Western blotting and RT-PCR. The results showed that the cellular melanin content was increased by CoPP and decreased by ZnPP. The Western blot and RT-PCR analyses showed that CoPP increased p53, MITF and tyrosinase levels, and ZnPP reduced all of them. The knockdown of p53 by siRNA transfection was followed by large decreases in the expression levels of p53, MITF and tyrosinase at 3 h of transfection. The presence of CoPP or ZnPP had no significant increased or decreased effects on MITF and tyrosinase levels from 15 h in the siRNA transfectants. Our results suggest that HO-1 modulates melanogenesis in human melanocytes via a p53-dependent pathway.
Subject(s)
Humans , Blotting, Western , Cobalt , Heme Oxygenase-1 , Heme , Melanins , Melanocytes , Microphthalmia-Associated Transcription Factor , Monophenol Monooxygenase , RNA, Small Interfering , Transfection , ZincABSTRACT
As a key regulator of melanogenesis, p53 controls microphthalmia-associated transcription factor (MITF) and tyrosinase expression. The anti-oxidant enzyme heme oxygenase-1 (HO-1) is induced by various forms of cellular stress and diverse oxidative stimuli. However, few studies have examined the role of HO-1 in melanogenesis. Therefore, the aim of this study was to determine the role of HO-1 in melanogenesis and the mechanism underlying this relationship. Cultures of normal human melanocytes were treated with the HO-1 inducer cobalt protoporphyrin (CoPP) or the HO-1 inhibitor zinc protoporphyrin (ZnPP). We then measured the melanin content of the cells. Additional analyses consisted of Western blotting and RT-PCR. The results showed that the cellular melanin content was increased by CoPP and decreased by ZnPP. The Western blot and RT-PCR analyses showed that CoPP increased p53, MITF and tyrosinase levels, and ZnPP reduced all of them. The knockdown of p53 by siRNA transfection was followed by large decreases in the expression levels of p53, MITF and tyrosinase at 3 h of transfection. The presence of CoPP or ZnPP had no significant increased or decreased effects on MITF and tyrosinase levels from 15 h in the siRNA transfectants. Our results suggest that HO-1 modulates melanogenesis in human melanocytes via a p53-dependent pathway.
Subject(s)
Humans , Blotting, Western , Cobalt , Heme Oxygenase-1 , Heme , Melanins , Melanocytes , Microphthalmia-Associated Transcription Factor , Monophenol Monooxygenase , RNA, Small Interfering , Transfection , ZincABSTRACT
Aims: This study reports on In vitro investigation of photodynamic antimicrobial activity of protoporphyrin IX (PPIX) in the presence and absence of Hydrogen peroxide (H2O2) against S. aureus and P. aeruginosa. Place and Duration of Study: Department of Medical Physics, Anna University, Chennai between December 2013 and February 2014. Methodology: A light-emitting diode (LED) was used as a light source to irradiate PPIX. The antibacterial effect was analyzed by standard plate counting method. Steady-state fluorescence spectroscopy technique was used to monitor the damage at protein level. Results: We found that the antibacterial effect is dependent on PPIX concentration as well as H2O2 concentration and light dose. PPIX-H2O2 combination showed higher bacterial reduction of 6.5 log10 and 2.7 log10 for S. aureus and P. aeruginosa respectively, when the light dose increased to 70 J/cm2. Fluorescence spectroscopic characterization showed a considerable change in the intensity of emission of tryptophan present in the microorganisms between pre- and post- APDT. Conclusion: PPIX-H2O2 is a promising combination for APDT against Gram positive and Gram negative bacteria. The LED seems to be a very good option for PDT because of its low cost and miniature in size.
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Objective To investigate the role of Heme oxygenase-1 in the effect of hyperbaric oxygen preconditioning (HBOP) against the brain edema formation after experimental intracerebral hemorrhage in rats.Methods The study was carried out by animal experiment in two steps by using 54 Spradgue-Dawley rats weighting from 300-350 g.In the first step,rats were treated with HBOP (HBOP group,n =3) or with sham pre-conditioning (Sham pre-conditioning group,n =3).All the rats were sacrificed 24 h after the preconditioning,and basal ganglion of brain tissue was taken for detect HO-1 level by using western blot analysis.In the second step,rats were divided into 4 groups (n =12 in each group):HBOP +ZnPP group,in which rats had a micro-pump intra-peritoneally implanted containing a specific HO-1 inhibitor ZnPPⅨ (Zinc protoporphyrin IX,0.01 mg/kg),Sham pre-conditioning + Znpp group,HBOP + DMSO group,in which rats with a intra-peritoneal micro-pump containing 2 mL Dimethyl sulfoxide (DMSO,a solvent vehicle) and Sham pre-conditioning + DMSO group before HBOP.At 24 hours after the pre-conditioning,rats received an infusion of 100 μL autologous blood into the caudate nucleus to form a simulated intracerebrum hemorrhage (ICH),and were sacrificed 72 h later for brain water content measurements.All data were analyzed by using Stata 7.0 software and statistical analyses were carried out by two-tailed Student t test.Results Compared with the Sham pre-conditioning group,the HBOP group had significant higher level of HO-1.Compared with the Sham pre-conditioning + DMSO group,the HBOP + DMSO group had a significant lower level of water content in the ipsilateral basal ganglion [(81.4 ± 0.9) ％ vs.(82.6 ± 0.8) ％ (P ＜ 0.05)],however,peritoneal infusion of ZnPP Ⅸ before HBOP abolished HBOP-induced protection against brain edema formation after experimental ICH [(82.8 ± 0.9) ％ vs.(82.6 ± 0.7) ％ (P ＞ 0.05)].Conclusions Hyperbaric oxygen preconditioning attenuate brain edema formation after experimental ICH in rats,and this protection is attributed to the activation of HO-1.
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La disponibilidad adecuada de hierro (Fe) es esencial para el desarrollo humano y la salud en general. El Fe es un componente clave de las proteínas portadoras de oxígeno, tiene un papel fundamental en el metabolismo celular y es esencial para el crecimiento y diferenciación celular. La ingesta inadecuada de Fe en la dieta, las condiciones inflamatorias crónicas o agudas y numerosas patologías están asociadas con alteraciones en la homeostasis de este metal. La regulación estricta del metabolismo del Fe es necesaria pues el Fe libre es altamente tóxico y los seres humanos sólo pueden excretar pequeñas cantidades a través del sudor, la piel, el enterocito y eliminarlo por pérdidas en procesos normales y patológicos. El objetivo de este trabajo es analizar los algoritmos para la evaluación preliminar tanto de la deficiencia como de la sobrecarga de Fe, sobre la base de diferentes parámetros, algunos accesibles, de simple resolución y que pueden ser efectuados en todos los laboratorios de análisis clínicos. Entre ellos, se analizarán el hemograma con los Índices hematimétricos, Reticulocitos, Fe sérico, Capacidad Total de Fijación de Hierro (CTFH) para calcular el Índice de Saturación de Transferrina (ISTf) y también el dosaje de Ferritina (Ft), todas mediciones que integran el "estudio del estado del hierro". Asimismo, se exponen y se consideran otros marcadores de uso poco frecuente en este medio, como la Protoporfirina Eritrocitaria Libre (PEL), la Eritropoyetina (EPO), entre otras, que ayudan desde el laboratorio al diagnóstico de una anemia. En los casos de sospecha de una sobrecarga de Fe, si bien la confirmación diagnóstica se realiza por estudios genéticos, como estudio inicial se reafirma la evaluación del paciente por medio del "estudio del estado del hierro" y especialmente el dosaje de Fe sérico y del ISTf para seguimiento del tratamiento instaurado. En las últimas décadas, se han producido importantes conocimientos sobre el metabolismo del Fe que han permitido descubrir otras proteínas que intervienen en el transporte, absorción, reciclaje y balance del Fe plasmático. Entre estas, existen marcadores séricos que podrían sumarse a los algoritmos propuestos y ellos son el Receptor de Transferrina (RTf) y la Hepcidina (Hp). Como conclusión, se destaca la necesidad de medir más de un marcador del "estado del hierro" para establecer el diagnóstico de una deficiencia o de un exceso de Fe.
Adequate availability of iron (Fe) is essential for human development and overall health. Iron is a key component of the oxygen-carrying proteins, it has a fundamental role in cellular metabolism, and it is essential for cell growth and differentiation. Inadequate intake of Fe in the diet, chronic or acute inflammatory conditions and many diseases are associated with alterations in the homeostasis of this metal. Strict regulation of Fe metabolism is necessary because free Fe is highly toxic and humans can excrete only small amounts through sweat, skin, and enterocyte loss in normal and pathological processes. The objective of this work is to analyze algorithms for the preliminary assessment of both Fe deficiency and overload, based on different parameters, some simple resolution ones that can be performed in all clinical laboratories. Among them, CBC, Hematimetric Indices, Reticulocytes, serum Fe, Total Iron Binding Capacity (TIBC) will be considered to calculate Transferrin Saturation Index (TfSI) and Ferritin Dosage (Ft), all measurements being part of the "study of iron status." Other markers of less frequent use in our region will also be considered, such as Free Erythrocyte Protoporphyrin (FEP), and Erythropoietin (EPO), among others, that help, from the laboratory in the diagnosis of anemia. In cases of suspected Fe overload, although the diagnosis was confirmed by genetic studies performed as initial study, the patient assessment is reaffirmed through the "study of iron status" and especially serum Fe and TfSI dosage for monitoring treatment underway. In recent decades, important insights on Fe metabolism have yielded more knowledge on other proteins involved in the transport, absorption, recycling and plasmatic Fe balance. Among these, there are serum markers that could be added to the proposed algorithms, which are Transferrin Receptor (TfR) and Hepcidin (Hp). In conclusion, the need to measure more than one analyte of the "iron status" is highlighted in order to establish the diagnosis of Fe deficiency or excess.
A disponibilidade adequada de ferro (Fe) é essencial para o desenvolvimento humano e para a saúde em geral. O Fe é um componente fundamental das proteínas transportadoras de oxigénio, tem um papel fundamental no metabolismo celular, e é essencial para o crescimento e diferenciagäo celular. A ingestäo inadequada de Fe na dieta, as condigöes inflamatorias crónicas ou agudas e inúmeras doengas estäo associadas a alteragöes na homeostase deste metal. A regulagäo rigorosa do metabolismo do Fe é necessària porque o Fe livre é altamente tóxico e os seres humanos apenas podem excretar pequenas quantidades através do suor, pele, enterócitos e eliminà-lo por perdas em processos normais e patológicos. O objectivo deste trabalho é analisar algoritmos para a avaliagäo prévia tanto da deficiéncia quanto do excesso de Fe, com base em diferentes parámetros, alguns acessíveis, de simples resolugäo e que podem ser realizados em todos os laboratorios clínicos. Dentre eles seräo analisados o hemograma com Índices hematimétricos, Reticulócitos, Fe sérico, Capacidade Total de Fixagäo do Ferro (CTFF) para calcular o Índice de Saturagäo da Transferrina (IST) e também a dosagem de Ferritina (Ft), todas elas medigóes que integram o "estudo do estado do ferro". Também sào expostos e considerados outros marcadores de uso pouco frequente nesse meio, como a Protoporfirina Eritrocitària Livre (PEL), a Eritropoietina (EPO), dentre outros, que ajudam a partir do laboratório ao diagnóstico de uma anemia. Nos casos de suspeita de um excesso de Fe, embora o diagnóstico seja confirmado através de estudos genéticos, como estudo inicial é reafirmada a avaliagäo do paciente por meio do "estudo do estado do ferro" e especialmente a dosagem de Fe sérico e do IST para o seguimento do tratamento instaurado. Nas últimas décadas, houve importantes co-nhecimentos a respeito do metabolismo do Fe que permitiram descobrir outras proteínas envolvidas no transporte, absorgäo, reciclagem e balango do Fe plasmàtico. Dentre elas, hà marcadores séricos que poderiam se unir aos algoritmos propostos e eles säo o Receptor de Transferrina (Tf) e Hepcidina (Hp). Em conclusäo, destaca-se a necessidade de medir mais de um marcador do "estado do ferro", para estabelecer o diagnóstico de uma deficiéncia ou de um excesso de Fe.
Subject(s)
Humans , Iron Overload , Iron/analysis , Algorithms , Laboratory and Fieldwork Analytical Methods/methods , Biomarkers , Blood Cell Count , Clinical Laboratory Services , Clinical Laboratory Techniques/methods , Iron/metabolism , Quality ControlABSTRACT
Objective To explore prevention of cyclosporine A (CsA) combined with Cobalt protoporphyrin (CoPP) against murine graft versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods C57BL/6 (H-2Kb) mice were used as donors and BALB/c (H-2Kd) mice as recipients,which were randomly divided into 4 groups.The mice in total body irradiation group (TBI group) were lethally irradiated and injected intravenously with PBS; The mice in Allo-HSCT group (BS group) were lethally irradiated and injected intravenously with bone marrow cells and spleen cells; The mice in CsA intervention group (CsA group) were injected with CsA intraperitoneally after allo-HSCT; The mice in CsA combine with CoPP intervention group (combination group) received both CsA and CoPP intraperitoneally after alloHSCT.Recipients were monitored for condition,survival rate and weight.The liver,small intestine and skin in the recipients were gained and pathological changes of GVHD were assessed.The kidney was stained with Masson staining dye to observe the tissue fibrosis.The expression levels of renal HO-1 mRNA in the recipients were detected.Results In contrast to BS and CsA groups,GVHD degree in combination group was mild,with less reduction and quick recovery of weight.On the day 30 after HSCT,survival rate in BS group was 36.8％,and that in combination group and CsA group was 69.6％ and 53.5％ respectively (P＜0.05).In comparison with BS and CsA groups,pathological changes in combination group were mild,cellular edema and degeneration degree of the liver,small intestine and skin were slight,and few necrosis and infiltrated inflammatory cells were observed.Tubulointerstitial fibrosis hardly occurred in combination group,but it occurred in CsA group abundantly.As compared with BS group,the expression levels of HO-1 mRNA was increased in combination group,while decreased in CsA group (P＜0.05).Conclusion CsA combined with CoPP enhanced the protective effect of CsA against GVHD,moreover,CoPP could alleviate the side effects of CsA,which might be related with up-regulation of the expression levels of HO-1.
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BACKGROUND: As some parameters reflecting iron status were known to change with infection or inflammation, we examined the changes of these parameters in children with minor illnesses.METHODS: Hematologic tests were done in 42 young children with acute infection. Iron deficiency anemia (IDA) was defined as having Hb less than age-matched normal range, MCH <27 pg, and either Tfsat (transferrin saturation) <10% or TIBC >360 microg/dL. Iron deficiency (ID) was defined as having Hb equal or more than age matched normal low limit with MCH <27 pg, and either Tfsat <10% or TIBC >360 microg/dL. The others were classified as normal control (NC).RESULTS: The proportion of IDA, ID and NC were 16.6% (7/42), 33.3% (14/42) and 50.0% (21/42), respectively. Comparisons of means of Hb, MCV, MCH, and RDW between groups showed statistical difference in general, while levels of iron, ferritin and hs-CRP showed no statistical difference. Mean blood levels of zinc protoporphyrin (ZnPP) of IDA, ID and NC were 72.21 microg/dL, 57.02 microg/dL, and 45.62 microg/dL, respectively, but the difference was significant only between IDA and NC. ZnPP was inversely correlated with MCV (r=-0.518, P<0.01) and RDW (r=-0.640, P<0.01), but not with hs-CRP or ferritin.CONCLUSION: Combination of RBC indices with newly controlled Tfsat or TIBC can be available for an iron status assessment in children with minor infections. ZnPP levels in blood reflect some aspect of iron status, while ferritin and iron do not reflect it.
Subject(s)
Child , Humans , Anemia, Iron-Deficiency , Communicable Diseases , Erythrocyte Indices , Ferritins , Hematologic Tests , Inflammation , Iron , Protoporphyrins , Reference Values , ZincABSTRACT
Objective To explore the effects of Zinc Protoporphyrin and Heme on the expression of HO-1 in cochlear and the change of auditory brainstem response on diffuse traumatic brain injury model of rats.Methods Diffuse traumatic brain injury model of rats were established and randomly divided into thirteen groups.Then auditory brainstem response examination,light microscope,immunohistochemistry technique were used to evaluate the change of auditory brainstem response and the expression of HO-1 in cochlear.Results The differences of auditory brainstem response threshold and latency of wave between the experimental and the normal control group were obvious(P＜0.05).The expression of HO-1 in the control group was normal,whereas there were obvious changes of inner ear HO-1 expression in the traumatic groups.The grey value of HO-1 expression in trauma group,Znpp group and heme group was significantly associated with auditory function change(P＜0.05).Conclusion There were influence of Zinc Protoporphyrin and Hemeon the inner ear HO-1 expression and the change of auditory brainstem response with diffuse traumatic brain injury model of rats.The protective effect of heme on auditory function may be associated with the increased expression of HO-1.
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BACKGROUND: As some parameters reflecting iron status were known to change with infection or inflammation, we examined the changes of these parameters in children with minor illnesses. METHODS: Hematologic tests were done in 42 young children with acute infection. Iron deficiency anemia (IDA) was defined as having Hb less than age-matched normal range, MCH 360 microg/dL. Iron deficiency (ID) was defined as having Hb equal or more than age matched normal low limit with MCH 360 microg/dL. The others were classified as normal control (NC). RESULTS: The proportion of IDA, ID and NC were 16.6% (7/42), 33.3% (14/42) and 50.0% (21/42), respectively. Comparisons of means of Hb, MCV, MCH, and RDW between groups showed statistical difference in general, while levels of iron, ferritin and hs-CRP showed no statistical difference. Mean blood levels of zinc protoporphyrin (ZnPP) of IDA, ID and NC were 72.21 microg/dL, 57.02 microg/dL, and 45.62 microg/dL, respectively, but the difference was significant only between IDA and NC. ZnPP was inversely correlated with MCV (r=-0.518, P<0.01) and RDW (r=-0.640, P<0.01), but not with hs-CRP or ferritin. CONCLUSION: Combination of RBC indices with newly controlled Tfsat or TIBC can be available for an iron status assessment in children with minor infections. ZnPP levels in blood reflect some aspect of iron status, while ferritin and iron do not reflect it.
Subject(s)
Child , Humans , Anemia, Iron-Deficiency , Communicable Diseases , Erythrocyte Indices , Ferritins , Hematologic Tests , Inflammation , Iron , Protoporphyrins , Reference Values , ZincABSTRACT
In 1967, the problem of occupational lead exposure came to public attention in Korea. Since then, regular progress has been made in lowering workplace lead exposures, instituting new workplace controls, and implementing health examinations of exposed workers. Past serious lead poisoning episodes made it possible to introduce biological monitoring programs on a voluntary basis in high-lead-exposure facilities in Korea. Industry-specific occupational health services for lead workers in Korea during the last 22 years can be categorized into three phases. During the first phase (1988-1993), efforts were directed at increasing awareness among workers about the hazards of lead exposure, biological monitoring of blood zinc protoporphyrin began, and a respiratory protection program was introduced. During the second phase (1994-1997), a computerized health management system for lead workers was developed, blood-lead measurement was added to biologic monitoring, and engineering controls were introduced in the workplace to lower air-lead levels to comply with air-lead regulations. Finally, during the third phase (1998-present), a new biomarker, bone-lead measurement by X-ray fluorescence, was introduced. Bone-lead measurement proved to be useful for assessing body burden and to demonstrate past lead exposure in retired workers. Occupational health service practice for lead workers, including the industry-specific group occupational health system, has brought considerable success in the prevention of lead poisoning and in reducing the lead burden in Korean lead workers during the last several decades. The successful achievement of prevention of lead poisoning in Korea was a result of the combined efforts of lead workers, employers, relevant government agencies, and academic institutes.
Subject(s)
Academies and Institutes , Achievement , Body Burden , Environmental Monitoring , Fluorescence , Government Agencies , Korea , Lead Poisoning , Occupational Health , Occupational Health Services , Porphyrins , Protoporphyrins , Social Control, Formal , ZincABSTRACT
BACKGROUND: Glucose toxicity that is caused by chronic exposure to a high glucose concentration leads to islet dysfunction and induces apoptosis in pancreatic beta-cells. Heme oxygenase-1 (HO-1) has been identified as an anti-apoptotic and cytoprotective gene. The purpose of this study is to investigate whether HO-1 up-regulation when using metalloprotophyrin (cobalt protoporphyrin, CoPP) could protect pancreatic beta-cells from high glucose-induced apoptosis. METHODS: Reverse transcription-polymerase chain reaction was performed to analyze the CoPP-induced mRNA expression of HO-1. Cell viability of INS-1 cells cultured in the presence of CoPP was examined by acridine orange/propidium iodide staining. The generation of intracellular reactive oxygen species (ROS) was measured using flow cytometry. Glucose stimulated insulin secretion (GSIS) was determined following incubation with CoPP in different glucose concentrations. RESULTS: CoPP increased HO-1 mRNA expression in both a dose- and time-dependent manner. Overexpression of HO-1 inhibited caspase-3, and the number of dead cells in the presence of CoPP was significantly decreased when exposed to high glucose conditions (HG). CoPP also decreased the generation of intracellular ROS by 50% during 72 hours of culture with HG. However, decreased GSIS was not recovered even in the presence of CoPP. CONCLUSION: Our data suggest that CoPP-induced HO-1 up-regulation results in protection from high glucose-induced apoptosis in INS-1 cells; however, glucose stimulated insulin secretion is not restored.
Subject(s)
Apoptosis , Caspase 3 , Cell Survival , Diabetes Mellitus , Flow Cytometry , Glucose , Heme , Heme Oxygenase-1 , Insulin , Protoporphyrins , Reactive Oxygen Species , RNA, Messenger , Up-RegulationABSTRACT
A fluorometric analytical method was developed for quantification of protoporphyrin IX (PpIX) in skin samples and receptor phase solution after in vitro cutaneous penetration/permeation studies. Analytical conditions used were: excitation and emission wavelengths: 400 nm and 632 nm; bandwidth: 0.5 nm; excitation and emission slits: 10/10. PpIX was recovered from two different layers of skin, the stratum corneum (SC) and the epidermis plus dermis ([E+D]), by vortex homogenization, probe and bath sonication, using DMSO as an extraction solvent. The detection and quantification limits were 0.002 and 0.005 μg/mL, respectively. The assay was linear from 0.005 - 0.5 μg/mL. The within-day and between-day assay precision and accuracy in DMSO and receptor phase solution were each studied at the two concentration levels 0.04 and 0.2 μg/mL, and 0.01 and 0.08 μg/mL, respectively. The coefficients of variation and deviation from the theoretical values were lower than 5%. The skin recovery of PpIX from SC and [E+D] layers using two different concentrations (0.5 and 1.0 μg/mL) were all above 90.0%. The method described has potential application to in vitro penetration/permeation studies of PpIX using porcine skin as a biological membrane model.
Um método analítico por espectrofluorimetria foi desenvolvido para quantificar a protoporfirina IX (Pp IX) em amostras de pele e fase receptora após a realização de testes in vitro de penetração/permeação cutâneas. As condições analíticas utilizadas foram: comprimentos de onda de excitação e emissão: 400 nm e 632 nm; largura de banda: 0,5 nm; fendas de excitação e emissão: 10/10. A PpIX foi extraída de amostras de estrato córneo (EC) e da epiderme sem estrato córneo + derme ([E+D]) através da agitação em vórtex e sonicação por haste e banho, utilizando-se o DMSO como solvente extrator. O limite de detecção e quantificação foram, respectivamente, de 0,002 e 0,005 μg/mL. O método mostrou-se linear da faixa de 0,005 - 0,5 μg/mL. A precisão e exatidão intra e inter-ensaio em DMSO e na fase receptora foram validadas utilizando-se duas concentrações distintas, respectivamente, de 0,004 e 0,2 μg/mL, e 0,01 e 0,08 μg/mL. Os valores de coeficiente de variação e o desvio do valor teórico foram inferiores a 5%. A recuperação da PpIX das camadas da pele (EC e [E+D]) utilizando-se duas concentrações distintas (0,5 e 1,0 μg/mL) foram todas acima de 90,0%. O método descrito pode ser utilizado para determinação da PpIX após estudos de penetração/permeação cutânea in vitro utilizando pele de porco como modelo de membrana.