2 Cases of Beta-thalassemia Minor in Korea / 임상소아혈액종양

In Korea, recent epidemiologic studies show that the incidence of β-thalassemia is increasing as the influx of South-East Asian population increases and molecular technologies develop. However, many patients are still misdiagnosed as iron deficiency anemia (IDA). All patients with microcytic anemia need to perform evaluation including reticulocyte index, Mentzer index, and iron studies. Considering the increasing incidence of β-thalassemia, hemoglobin beta globulin (HBB) gene sequencing should be performed if suspicious. In our cases, patients whose parents were both Koreans were confirmed to have β-thalassemia with a substitution in c1, ATG>GTG, and deletion of the HBB gene. In Korea, initiation condon ATG>AGG (20.9%) is most common mutation, followed by codon 17 (A>T) (17.6%), codon 121 (G>T) (12.1%), and so on. We report two cases of β-thalassemia diagnosed by genetic testing for microcytic anemia.

2 Cases of Beta-thalassemia Minor in Korea / 임상소아혈액종양

In Korea, recent epidemiologic studies show that the incidence of β-thalassemia is increasing as the influx of South-East Asian population increases and molecular technologies develop. However, many patients are still misdiagnosed as iron deficiency anemia (IDA). All patients with microcytic anemia need to perform evaluation including reticulocyte index, Mentzer index, and iron studies. Considering the increasing incidence of β-thalassemia, hemoglobin beta globulin (HBB) gene sequencing should be performed if suspicious. In our cases, patients whose parents were both Koreans were confirmed to have β-thalassemia with a substitution in c1, ATG>GTG, and deletion of the HBB gene. In Korea, initiation condon ATG>AGG (20.9%) is most common mutation, followed by codon 17 (A>T) (17.6%), codon 121 (G>T) (12.1%), and so on. We report two cases of β-thalassemia diagnosed by genetic testing for microcytic anemia.

Clinical observation of the γ-globulin levels when Benign paroxysmal positional vertigo is attacking / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the characteristics of serum protein in patients with benign paroxysmal positional vertigo (BPPV) during the symptomatic period.@*METHOD@#Fifty-five patients with BPPV and 58 control subjects were enrolled in the study. All the patients underwent the Dixe-Hallpike and Roll maneuver to confirm the type of BPPV. The average time of onset was (1.0 ± 0.8)days in the group of BPPV. The clinical features and the laboratory tests of serum protein electrophoresis, blood counts, and liver and kidney function were performed in both groups.@*RESULT@#The levels of serum albumin (Alb), α1 globulin, α2 globulin and β globulin of BPPV group did not differ statistically in the two groups (P > 0.05); The γ-globulin levels were significantly higher in patients with BPPV than in controls (P < 0.05).@*CONCLUSION@#The γ-globulin levels are increased when BPPV is attacking.

Molecular analysis and association with clinical and laboratory manifestations in children with sickle cell anemia

Objectives: To analyze the frequency of βS-globin haplotypes and alpha-thalassemia, and their influence on clinical manifestations and the hematological profile of children with sickle cell anemia. Method: The frequency of βS-globin haplotypes and alpha-thalassemia and any association with clinical and laboratorial manifestations were determined in 117 sickle cell anemia children aged 3–71 months. The confirmation of hemoglobin SS and determination of the haplotypes were achieved by polymerase chain reaction-restriction fragment length polymorphism, and alpha-thalassemia genotyping was by multiplex polymerase chain reaction (single-tube multiplex-polymerase chain reaction). Results: The genotype distribution of haplotypes was 43 (36.7%) Central African Republic/Benin, 41 (35.0%) Central African Republic/Central African Republic, 20 (17.0%) Rare/atypical, and 13 (11.1%) Benin/Benin. The frequency of the α3.7 deletion was 1.71% as homozygous (−α3.7/−α3.7) and 11.9% as heterozygous (−α3.7/αα). The only significant association in respect to haplotypes was related to the mean corpuscular volume. The presence of alpha-thalassemia was significantly associated to decreases in mean corpuscular volume, mean corpuscular hemoglobin and reticulocyte count and to an increase in the red blood cell count. There were no significant associations of βS-globin haplotypes and alpha-thalassemia with clinical manifestations. Conclusions: In the study population, the frequency of alpha-thalassemia was similar to published data in Brazil with the Central African Republic haplotype being the most common, followed by the Benin haplotype. βS-globin haplotypes and interaction between alpha-thalassemia and sickle cell anemia did not influence fetal hemoglobin concentrations or the number of clinical manifestations...

A case of Fanconi syndrome accompanied by crystal depositions in tubular cells in a patient with multiple myeloma

Fanconi syndrome (FS) is a rare condition that is characterized by defects in the proximal tubular function. A 48-year-old woman was admitted for evaluation of proteinuria. The patient showed normal anion gap acidosis, normoglycemic glycosuria, hypophosphatemia, and hypouricemia. Thus, her condition was compatible with FS. The M peak was found behind the beta globulin region in urine protein electrophoresis. Upon bone marrow examination, we found that 24% of cells were CD138+ plasma cells with kappa restriction. From a kidney biopsy, we found crystalline inclusions within proximal tubular epithelial cells. Thereafter, she was diagnosed with FS accompanied by multiple myeloma. The patient received chemotherapy and autologous stem cell transplantation, and obtained very good partial hematologic response. However, proximal tubular dysfunction was persistent until 1 year after autologous stem cell transplantation. In short, we report a case of FS accompanied by multiple myeloma, demonstrating crystalline inclusion in proximal tubular cells on kidney biopsy.

Effects of modified liangge powder contained serum on LPS stimulated TLR4 expression and release of cytokines in mouse platelets / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of Modified Liangge Powder (MLP) on the expressions of platelet toll like receptor 4 (TLR4) and the release of platelet-derived cytokines interleukin 8 (IL-8), beta platelet globulin (beta-TG), soluble CD40 ligand (sCD40L).</p><p><b>METHODS</b>The modulating effects on the release of cytokines from mice platelets by TLR4 ligand through monoclonal antibody blocking TLR4 on platelet were compared. The stimulated platelet by LPS was incubated with low (0.94 g/mL), medium (1.89 g/mL), and high (2.84 g/mL) dose of MLP contained serum. The changes of the platelet TLR4 expression and platelet-derived cytokines were observed.</p><p><b>RESULTS</b>The positive expression rate of platelet TLR4 obviously decreased (P < 0.01) and the release of sCD40L and beta-TG from platelets significantly increased (P < 0.01) after stimulated by LPS. However, the release of sCD40L and beta-TG from platelets obviously decreased by TLR4 monoclonal antibody (P < 0.05, P < 0.01). There was no statistical difference in IL-8 between before and after LPS stimulation (P > 0.05). Platelet TLR4 positive expression rate was significantly higher after incubated by medium and high doses of MLP contained serum (P < 0.01), and the releasing of sCD40L and beta-TG was lower in the serum contained groups. The inhibitory effects were enhanced in a dose-dependent manner.</p><p><b>CONCLUSIONS</b>LPS induced platelet activation by TLR4 and released sCD40L and beta-TG, while the release of platelet IL-8 was not dependent on platelet TLR4-LPS pathway. MLP could inhibit LPS-stimulated sCD40L and beta-TG, inhibit the binding of platelet TLR4 and LPS in a dose-dependent manner, thus reducing the release of platelet cytokines.</p>

Perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro / Serum protein electrophoresis profile of the rattlesnake Crotalus durissus terrificus kept in captivity

As serpentes peçonhentas dos gêneros Bothrops e Crotalus têm sido mantidas em cativeiro visando à extração de venenos para a produção de imunobiológicos. O conhecimento da fisiologia desses animais e as alterações na concentração de proteínas e suas frações séricas são importantes para a identificação precoce de importantes enfermidades que cursam com estados de hipoproteinemia e hiperproteinemia. O objetivo do trabalho foi determinar a concentração de proteína total e o perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro. Foram colhidas amostras de sangue da veia coccígea ventral de 21 serpentes adultas e sadias, divididas em dois grupos: Grupo 1 de 12 machos com peso médio de 588,89±193,55g, e Grupo 2 de nove fêmeas com peso médio de 708,33±194,04g. A proteína total sérica foi determinada pelo método de refratometria e a eletroforese em gel de agarose. Obtiveram-se valores da proteína total sérica (g/dL) de 4,51±0,50 para machos e de 4,82±0,72 para fêmeas, e para machos e fêmeas de 4,64±0,61. Foram identificadas pela eletroforese quatro frações protéicas (g/dL): albumina, a, b, g-globulinas e calculada a relação albumina:globulina. As serpentes fêmeas apresentaram maiores valores para as variáveis, albumina e para a relação albumina/globulina (AG) diferindo significativamente (P<0,05) do grupo de machos, porém sem significado clínico.

The poisonous snakes of the genera Crotalus and Bothrops have been kept in captivity with the purpose of extracting poison for the production of immunobiological. Knowledge of the physiology of these animals and serum proteins concentration changes are important for early identification of major diseases which lead to states of hypoproteinemia and hyperproteinemia. The objective was to determine the concentration of total protein and serum protein electrophoresis profile of Crotalus durissus terrificus (rattlesnake) in captivity. Blood samples were taken from the ventral coccygeal vein of 21 adult and healthy snakes divided into groups: Group 1 with 12 males, weighing in average 588.89±193.55g, and Group 2 with nine females, weighing in average 708.33±194.04g. The total serum concentration of protein was determined by the method of refractometry and agarose gel electrophoresis. The total protein values in the serum for females was 4.82±0.72, for males 4.51±0.50 and males and females 4.64±0.61, identified by four fractions (g/dL): albumin, a, b and g-globulin. Additionally the albumin/globulin ratio was calculated. The female snakes showed higher values for the variables, albumin and the albumin/globulin (AG) differed significantly (P<0.05) from the group of male snakes, but there was no clinical significance.

[Effect of saffron extract on electrophoretic pattern of serum proteins in the male mice]

Due to widespread use of saffron [Crocus sativus L] as food colorant and flavor, and its reputation in folk medicine as a drug, recent studies revealed that main components of saffron are the carotenoids: crocin, crocetin, picrocrocin and safranal which have a large number of physiological effects on different biological systems. Our objective was to assess the efficacy of Crocus sativus on serum proteins pattern in the male mice. Five groups including eight adult male Balb/C mice were used in this study. Normal saline administered as placebo to control group and saffron extract in doses of 25 mg/Kg/48hr, 50 mg/Kg/48hr, 100 mg/Kg/48hr and 200 mg/Kg/48hr were injected intra peritoneally for 20 days to experimental groups. The levels of Albumin, Alpha-1, Alpha-2, Beta and Gamma globulins were separated electrophoretically and A/G ratio was calculated from the pattern of electrophoretogram. The result indicated that the levels of Albumin increased significantly in two experimental groups that had received 50 mg/Kg/48h and 100 mg/Kg/48h extract of Saffron as compared to the control group, the levels of Alpha-1 didn't have any remarkable changes in any group. The injection of 50 mg/Kg/48h, 100 mg/Kg/48h and 200 mg/Kg/48hr extract of saffron decreased [p < 0.05] the Alpha-2 level in plasma as compared to the control group and levels of Beta globulins increased significantly in these three groups. The levels of Gamma globulins increased significantly in 100 and 200 mg/kg-treated groups as compared to the placebo controlled group. A/G ratio [Albumin/Globulin ratio] were significantly [p < 0.05] lower than control group in any groups that received saffron extract in a dose-dependent manner. Albumin were significantly increased in two groups and A/G ratio was decreased in any groups. This can be interpreted that in the absence of antigen stimulation, serum globulins did alter markedly by extract of saffron. The study shows that since albumin synthesis occurs in the liver cells, thus administration of saffron may improve the status of liver function significantly

Plasma level of von Willebrand factor: an indicator of severity in sickle cell disease

Sickle cell anaemia is a congenital hemolytic disorder caused by mutation in the beta-globin gene at position 6 with replacement of glutamic acid by valine. Patients who are homozygous for this mutation suffer from hemolytic anaemia and other serious complications. The underlying pathology of many of these complications is the occurrence of recurrent vasoocclusion due to microthrombi formation resulting in organs ischaemia. In this study we investigated the role of vWF as a determinant of sickle cell disease severity through its contribution to the formation of such microthrombi. The clinical disease severity was determined using two different scoring methods, and vWF antigen level in the plasma was estimated by using ELISA technique. Seventy Sudanese patients were investigated in this study. They were 35 females and 35 males, the mean of their ages +/- standard deviation was 6.8 +/- 4.7 years. Seventeen controls with normal haemoglobin were also included with mean age of 6.5 years. Thirty four patients presented during vasoocclusive crisis and 36 presented in steady state. vWF was high [179.83%] among patients compared to controls [82.4%], p< 0.001 there was positive correlation between severity score and vWF level in the plasma in steady state in the two scoring methods used [r = 0.79, p = 0.008 for method I and r = 0.78, p = 0.009 for method II]. So, severity of sickle cell disease increases with elevation of vWF level in the plasma in the steady state

Characterization of beta globin gene mutations in Zanjan province: An introduction to prenatal diagnosis of Thalassemia

B-thalassemia is an autosomal recessive disease characterized by reduction or complete absence of b-globin gene expression. It has been estimated that more than 2,000,000 carriers as well as 20,000 patients affected with b-thalassemia are living in Iran, a country with more than 70 million population and great ethnic diversity. In this study we aimed to find out the b-globin gene frequency and determine the spectrum of b-globin gene mutations in Zanjan province [northwest region] of Iran. 5527 individuals who were referred for pre-marriage tests to Zanjan clinic as well as 27 thalassemia patients were studied. Altogether one hundred and five chromosomes from 78 unrelated Bthalassemia patients or carriers were examined for b-globin gene mutations by ARMS-PCR and direct gene sequencing. Based on the previous information on common mutations in Mediterranean populations 24 sites were analyzed. It was found that the b-thalassemia frequency is 1.2 for Zanjan region. Using the above techniques, the mutations for 90/105 [86.7] of b-thalassemia chromosomes [13 different mutations] were identified. Fifty eight percent of the mutations were of common .Mediterranean. type. Of which, IVS-I 110 mutation showed the highest frequency [29.5] followed by IVS-II-1 [13.3], IVS-I-1 [12.4] and IVS-I-6 [2.9]. 10.5% of mutations were of common Asian Indian mutations [Fr 8/9, 6.7% and IVS-I-5, 3.8] respectively. CD5 and CD30 and CD36-37 mutations accounted for 13.3% of the mutations. [5.7%, 0.95% and 6.7% respectively] Mutations in 14 chromosomes [13.3] remained uncharacterized. These data suggests that the spectrum of mutations in Zanjan province differs from those reported from other parts of Iran, but Mediterranean type of mutations are more frequent in Zanjan region. Therefore, in order to save the time and cost, it is recommended that for prenatal diagnosis of thalassemia in Zanjan province analysis of Mediterranean mutations should be considered as a front line screening strategy

Transduction of COS-7 and K562 cell lines by recombinant lentiviral particles carrying miniLCR and B-globin gene: Introduction to gene therapy of major B-thalassemia

Beta-thalassemia is caused by absence of reduction of beta-globin chain synthesis. One of the effective therapeutic methods for this disease can be gene therapy by viral vectors. The capacity of lentiviral vectors is approximately 8 kb, we designed a 6 kb construct containing mini LCR and beta-globin gene instead of LCR region. The aim of this study is to make a recombinant lentiviruses containing miniLCR and beta-globin gene for transfer to the target cells for gene therapy of beta-thalassemia. HS2, HS3, HS4 segments [miniLCR] and beta-globin gene with 5' and 3' UTR were amplified from the genomic DNA of a normal individual by PCR. Each segment was cloned in pTZ57R/T vector and then sub cloned first into the pBGGT vector and finally into the pLenti-Dest vector. Final transfer vector and the three helper packaging plasmids [Plp1, Plp2 Plp/VSVG] were contransfected into 293T packaging cells using lipofectamine 2000. Harvested viruses were confirmed by RT-PCR on extracted RNA of these recombinant lentiviruses. The titer of lentiviral stock determined in a K562 cell line and compared with COS-7 cell line. The titer in both cell lines was the same. Optimum MOI for COS-7 cell line was 5 and when polybrene was used transduction increased by 2 fold. The remaining transduced COS-7 colonies were expanded and DNA was extracted. By PCR, random intergration construct into the genome was evaluated. The produced lentiviruses can be an appropriate means for effective transfer of the designed construct into dividing and non-dividing cells such as hematopoetic stem cells for transplantation of beta thalassemia patients. Efficiency of transduction by leniviruses is more than the gene targeting technique. Also units of HS2, HS3 and HS4 regions in mini LCR and selection of larger HS3 unit may increase the expression of beta globin gene

The effects of high-dose intravenous immunoglobulin on plasma protein and lipid levels in the patients with Kawasaki disease / 소아과

PURPOSE: The reticuloendothelial system is composed of sinusoidal capillaries, through which even large protein molecules are freely movable between plasma and interstitial space, including the lymphatic system. Therefore, high-dose intravenous immunoglobulin (IVIG) would cause a redistribution of proteins between two compartments. To investigate this hypothesis, we measured plasma protein and lipid levels in patients with Kawasaki disease before and after high-dose IVIG treatment. METHODS: Thirty four children with Kawasaki disease who had complete responses to high-dose IVIG treatment (1 g/kg/day for two consecutive days), were analyzed. Before and after the administration of IVIG, serum analyses were performed for such parameters as total protein, albumin, gamma-globulins (IgG, IgM, IgA), alpha1-, alpha2-, and beta-globulin fractions, and lipid profiles (total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride). RESULTS: The levels of gamma-globulins including IgG, IgM, IgA were significantly increased, and IgG was increased by 1,779+/-304 mg/dL after two-dose of IVIG infusion. The levels of albumin, alpha1-, alpha2-, and beta-globulin fractions were significantly decreased by 18 percent, 24 percent, 19 percent and 12 percent, respectively. HDL-cholesterol level was significantly decreased by 20 percent, while LDL-cholesterol and triglyceride levels were significantly increased by 21 percent and 50 percent, respectively. The total cholesterol level was not changed. CONCLUSION: High-dose IVIG treatment decreased the levels of a variety of proteins except immunoglobulins, and the increase of IgG after IVIG treatment was lower than expected. Our results suggest that a part of infused IVIG and plasma proteins, including etiologic proteins for Kawasaki disease, may be distributed to the extravascular compartments. The rapid improvement of symptoms induced by IVIG in Kawasaki disease might be explained by this mode of action of IVIG.

Clinical phenotypes and molecular diagnosis in a hitherto interaction of Hb E/beta thalassemia syndrome (beta(E)/beta(-31), (A -->G)).

Molecular identification of affected alleles in the index family with rare mutation(s) and/or interaction(s) is an important prerequisite toward a proper genetic counseling. In Thailand, where more than 30% of the populations are heterozygotes for either alpha or beta thalassemia mutation(s). More than 60 different thalassemia syndromes resulting from the interactions of these heterogeneous alleles have been observed. The majority of patients in the hospital based-study are compound heterozygotes for beta thalassemia alleles and another hemoglobinopathy namely Hb E, highly prevalent in Thailand, gave rise to Hb E/beta thalassemia syndrome. The phenotypes of these syndromes vary from asymptomatic individual to a very severe phenotype mimic that of beta thalassemia major. In this report, we describe a three-year-old Thai girl presenting with mild hypochromic microcytic anemia since birth. She was born prematurely and developed anemia within the first week of life. The cause of anemia was suspected to result from prematurity and low intrauterine iron storage, however hypochromic anemia did not resolve after a three-month of iron supplement therapy. Subsequent studies indicated that the patient had Hb E/beta thalassemia disease and the molecular study revealed that the patient was a compound heterozygote for Hb E and a rare beta thalassemia mutation (beta(-31), A --> G). This hitherto genotype results in a relatively mild clinical symptom since the patient's baseline Hb values were around 9-10 g/dL with normal weight and height development during the follow-up period.

Urinary ?-microglobulin excretion for detecting tubular dysfunction in adult patients with primary nephrotic syndrome

The research studied urinary beta2-microglobulin excretion to detect tubular dysfunction in 31 patients with primary nephrotic syndrome. The result showed the increased excretion of beta2-microglobulin was observed in 11/31 cases. There were no sighgicant differences of the urinary creatinine excreation and between group of patients with increased urinary beta2-microglobulin excretion and the group without it. This indicated having combine tubular dysfunction in patients with primary nephrotic syndrome. The finding might suggest the use of urinary beta2-microglobulin for detecting the associated tubular disfuntion in such patients

Genetic variants of beta-globin gene in Thai malaria patients.

Hemoglobin E (E26K variant of beta-globin gene) causing hemoglobinopathy is commonly observed in parts of Thailand, regardless of the hematologic disadvantage of the homozygotes. In order to detect further variants of the beta-globin gene, we performed variation screening for exon 1 of the beta-globin gene in 64 adult patients with P. falciparum malaria, living in northwest Thailand. We identified E26K and two novel variants, 59C>T and IVS+1G>T. IVS+1G>T lies on the splice donor site, and a substitution of A for G at the same site (IVS+1G>A) is known to be linked to beta-thalassemia. Thus, the biological significance of IVS+1G>T and its association with malarial infection should be clarified in future studies.

Mieloma múltiplo com pico monoclonal na fraçäo beta / Multiple myeloma with a monoclonal peak at beta-globulin fraction: case report

É relatado o caso de um paciente idoso, com quadro agudo de dorsalgia e dasabamento da sétima vértebra torácica, cuja eletroforese de proteínas séricas demonstrava um pico monoclonal na fraçäo beta-globulinas, achado pouco freqüente, encontrado em apenas 10 por cento dos casos de mieloma múltiplo. O paciente foi submetido a uma punçäo aspirativa de medula óssea via esternal. No mielograma observou-se 70 por cento de plasmócitos isolados e agregados, alguns displásicos e binucleados, compatível com mieloma múltiplo. Iniciou-se quimioterapia em janeiro de 2000, com Alkeran e Prednisona, tendo o paciente rapidamente apresentado melhora clínico-laboratorial.

Reliability of some biochemical parameters as prognostic factors for hepatitis in schistosomiasis patients

The early diagnosis of hepatitis in patients is sometimes difficult and despite adequate studies may remain undetected. To carry out this point, 4 selected groups of patients were investigated. Group 1 included patients with chronic liver cirrhosis due to schistosomiasis with a positive ELISA and PCR tests for HCV and showed a very high level of iron and ferritin as compared with the control group. Group 2 consisted of patients with chronic liver cirrhosis due to schistosomiasis with negative ELISA for either HBV or HCV antibodies and revealed a significant increase in copper and decrease in zinc as compared with the control group. Group 3 composed of patients with HCV infection, confirmed by antibody detection through third generation ELISA, RIBA III. Patients in this group showed a significant increase in the iron and ferritin level compared with the control group. Moreover, group 4 encompassed patients with HBV infection showed a positive ELISA for antibody detection [HBsAb] and immunochromatographic reaction for detection of surface antigen [HBsAg]. The results suggested that reduced level of Zn ++ and increased level Cu ++ might be a reflection of the incidence of HBV more than HCV infection, while a high level of iron and ferritin are potential biomarkers for schistosomiasis patients with early HCV infection

Serum protein concentrations and electrophoretic patterns by categories of small opacity profusions in coal workers' pneumoconioses / 예방의학회지

The concentrations of serum proteins fractions and their electrophoretical patterns were investigated in 135 patients with coal workers' pneumoconiosis who participated in confirmative examination for pneumoconiosis in December 1989. Their radiographical profusions were classified as 1/0 or more. Agarose film and phosphoric acid-sodium hydroxide buffer(pH 8.6) were used for electrophoresis. Concentration of each protein fractions and electrophoretical patterns seemed to be equivalent to reference values. Serum alpah1- and beta-globulin concentrations, however, were significantly different(p<0.50) among categories of small opacity profusions and showed the lowest level in the group of category 1. Albumin concentrations decreased and alpha2-globulin concentrations increased significantly(p<0.05) in the group of complicated with pulmonary tuberculosis. gamma-globulin concentrations were not varied by category of profusions nor by pulmonary tuberculosis complication.