Research progress of immunotherapies on correction of immunoparalysis in sepsis / 药学实践杂志

Immunoparalysis is the main cause of death in patients with intermediate and terminal sepsis. The correction of immunoparalysis is an important direction of sepsis treatment. In the pathological process of sepsis, a variety of factors contribute to the imbalanced secretion of cytokines, weakened function of antigen-presenting cells, apoptosis and depletion of lymphocytes, and ultimately lead to immunoparalysis, secondary infection, and even patient deaths. Cytokines such as GM-CSF, IFN-γ, IL-7, and IL-15, immune checkpoint-related therapies such as PD-1/PD-L1 antibodies, CTLA-4 antibodies, TIM-3 antibodies, and LAG-3 antibodies, and immunoreactive substances such as thymosin α1 and immunoglobulin might be beneficial to correct the immune paralysis of patients. the progress of immunotherapy to correct immune paralysis in sepsis were reviewed in this article.

Differences in Newborn Screening Results Among Women with Gestational Diabetes Mellitus

Abstract Multiple studies undertaken on cord blood demonstrate analyte perturbations in infants exposed to gestational diabetes mellitus (GDM). Cord blood as a sample is influenced by maternal and placental metabolism. Newborn screening (NBS), performed after the first 24 hours of life reflects early neonatal metabolism. We compared NBS analytes between women with and without GDM with different management approaches in the Treatment of Booking of Gestational Diabetes (TOBOGM) pilot randomised controlled trial. Pregnant women with GDM risk factors were randomised to early or deferred GDM treatment following an oral glucose tolerance test (<20 weeks gestation). Women without GDM served as "decoys". From the decoy group 11 developed GDM (screened at 26-28 weeks), were analysed separately; their results were compared with the other groups. De-identified controls were chosen from NBS results from the same analytic run matched for sex, birthweight and gestational age. Results were available for 73/78 women participating in the pilot and 358 de-identified controls. Tyrosine levels (μmol/l; whole blood)were higher in the late GDM group vs early, deferred treatment, and decoy groups (medians:106.28; IQR: 96.73-151.11) (76.33; 64.64-97.90) (75.68; 66.59-110.88)(73.74; 58.32-90.36) (p=0.009) and remained elevated when compared to normal, age-matched controls (106.28; 96.73-151.11) (87.26; 68.55-111.26) (p value=0.01) Immunoreactive trypsinogen (μgm/l; whole blood)was highest in the early treatment group when compared with group-specific controls (22.30; 13.90-29.90 vs 14.00, 10.60-21.10) (p=0.02). These results provide evidence of biochemical perturbations detectable on NBS of in-utero exposure to hyperglycemia and treatment and provide data for hypothesis building.

Proteomic analysis of excretory-secretory products from young adults of Angiostrongylus cantonensis

BACKGROUND Angiostrongyliasis is caused by the nematode Angiostrongylus cantonensis and can lead to eosinophilic meningitis and meningoencephalitis in humans. The young adult worms play central pathogenic roles in the central nervous system (CNS); however, the underlying mechanism is unclear. Excretory-secretory products (ESPs) are good investigation targets for studying the relationship between a host and its parasite. OBJECTIVES We aimed to profile, identify, and characterise the proteins in the ESPs of A. cantonensis young adults. METHODS The ESPs of young adult worms were collected from culture medium after incubation ranging from 24 to 96 h. Proteomic and bioinformatics analyses were performed to characterise the ESPs. FINDINGS A total of 51 spots were identified, and the highly expressed proteins included two protein disulphide isomerases, one calreticulin, and three uncharacterised proteins. Subsequently, approximately 254 proteins were identified in the ESPs of A. cantonensis young adults via liquid chromatography-mass spectrometry (LC-MS/MS) analysis, and these were further classified according to their characteristics and biological functions. Finally, we identified the immunoreactive proteins from a reference map of ESPs from A. cantonensis young adults. Approximately eight proteins were identified, including a protein disulphide isomerase, a putative aspartic protease, annexin, and five uncharacterised proteins. The study established and identified protein reference maps for the ESPs of A. cantonensis young adults. MAIN CONCLUSIONS The identified proteins may be potential targets for the development of diagnostic or therapeutic agents for human angiostrongyliasis.

Metodología para la obtención y evaluación de un extracto proteínico de una cepa autóctona de Helicobacter pylori / Methodology to obtain and evaluate a protein extract of an autochthonous Helicobacter pylori strain

Helicobacter pylori es una bacteria gram negativa que posee numerosos antígenos que juegan un importante papel en la patogénesis de las enfermedades gastroduodenales. Debido a la necesidad de métodos de diagnóstico estandarizados con antígenos locales u autóctonos nos propusimos el diseño de una estrategia para la obtención de extractos de antígenos con reactividad frente a sueros de pacientes infectados por H. pylori. Dos cepas de H. pylori, una autóctona (IPK196A) y una de referencia ATCC 43504, se cultivaron en un medio líquido modificado. Se sometieron a los protocolos de ruptura por ultrasonido aplicándose tres variantes de precipitación y al fraccionamiento celular mediante ultracentrifugación diferencial. Los extractos proteínicos se visualizaron mediante electroforesis en gel de poliacrilamida y se transfirieron para la detección de antígenos inmunorreactivos a sueros de pacientes con infección por H. pylori e individuos sanos. La variante de ultrasonido y precipitación con Coomasie fue la más efectiva para concentrar las muestras. El método de ultracentrifugación mejoró la resolución de las proteínas reactivas y permitió separarlas según su localización subcelular. El sistema de transferencia húmedo fue ideal para la inmunodetección de los antígenos obtenidos por ultrasonido mientras que el sistema semiseco permitió detectar las proteínas de membrana obtenidas por ultracentrifugación diferencial. La introducción de una metodología en el laboratorio para la obtención y evaluación de extractos proteínicos antigénicos a partir de cepas autóctonas de H. pylori, constituye la antesala para el diseño de futuros diagnosticadores y candidatos vacunales(AU)

Helicobacter pylori is a gram-negative spiral-shaped bacterium, which has many antigens that play an important role in the pathogenesis of gastroduodenal diseases. Due to the lack of standardized methods from native or autochthonous antigens, we proposed in this study, the design of a strategy for extracting and obtaining immunoreactive antigens against H. pylori infected-patient sera. Two H. pylori strains, one autochthonous (IPK196A) and one reference ATCC 43504, were cultured in a modified liquid medium. Both strains were subjected to the ultrasound rupture protocols applying three precipitation variants and cell fractionation by differential ultracentrifugation. Protein extracts were visualized by polyacrylamide gel electrophoresis and transferred for the detection of immunoreactive antigens to sera from patients with H. pylori infection and healthy individuals. The precipitation with Coomasie was the most effective variant. The ultracentrifugation extraction method optimized the resolution of the proteins, which could be separated according to their subcellular location. The wet transfer system was ideal for the immunodetection of the antigens obtained by ultrasound, while the semi-dry system allowed detecting the membrane proteins by differential ultracentrifugation. The introduction of a methodology in the laboratory for obtaining and evaluating antigenic antibodies from autochthonous strains of H. pylori, is the prelude to the design for future diagnostics and vaccine candidates(AU)

Effects of Sevelamer Carbonate Combined with Routine Treatment on Serum Inflammatory Factors, HO-1 and iPTH Levels of Chronic Renal Failure Patients with Hyperphosphatemia / 中国药房

OBJECTIVE: To investigate the effects of sevelamer carbonate combined with routine treatment on serum inflammatory factors, heme oxygenase-1 (HO-1) and intact parathyroid hormone (iPTH) levels of chronic renal failure (CRF) patients with hyperphosphatemia.METHODS: Totally 60 chronic renal failure patients with hyperphosphatemia in department of renal internal medicine, Tangshan Workers' Hospital during Jan. 2014 to Jan. 2015 were divided into control group and observation group according to random number table, with 30 cases in each group. Control group received routine treatment as reducing blood glucose, lowering blood lipid, lowering blood pressure, reducing phosphorus, protecting kidney. Observation group was additionally given sevelamer carbonate (dose was 0. 8 g, 3 times a day, during meal) on the basis of control group, for 4 weeks. The levels of IL-6, CRP, HO-1, iPTH, Ca and P were compared between 2 groups. The occurrence of ADR was compared between 2 groups. RESULTS: Two cases were withdrewn from the study in each group, finally 28 cases were included in the study in each group. Before treatment, there was no statistical significance in each index between 2 groups (P>0. 05). Compared with before treatment, the serum levels of IL-6, CRP, iPTH and P in 2 groups were decreased significantly after treatment (P<0. 05), while serum level of Ca was increased significantly (P<0. 05); the improvement of above indexes in observation group were more dovious than control group (P<0. 05). There was no statistical significance in the incidence of ADR between 2 groups (P>0. 05). CONCLUSIONS: Sevelamer carbonate combined with routine treatment can effectively reduce the serum levels of IL-6, CRP, HO-1 and iPTH and regulate Ca and P metabolism of CRF patients with hyperphosphatemia with good safety.

Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica / Detection of immunoreactive proteins of Rickettsia sp. Atlantic Forest strain

A Febre Maculosa Brasileira (FMB) é uma doença infecciosa, transmitida por carrapatos ao homem. Uma nova riquetsiose humana foi descrita como causadora de Febre Maculosa no Estado de São Paulo, sendo denominada de Rickettsia sp. cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune de hospedeiro mamífero, desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de Rickettsia sp. cepa Mata Atlântica. As proteínas extraídas foram fracionadas por eletroforese. As bandas proteicas foram transferidas para membranas de nitrocelulose por migração elétrica e submetidas à técnica de Western-blot, para detecção proteica. Ao todo sete proteínas imunorreativas foram detectadas. Duas proteínas apresentaram maior abundancia, com peso molecular, de 200 e 130 kDa respectivamente. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que as duas proteínas detectadas representem rOmpA (200 kDa) e rOmpB (130 kDa). As demais proteínas detectadas apresentaram menor ocorrência e peso molecular inferior a 78 kDa, podendo representar membros da família de antígenos de superfície celular (Sca - Surface cell antigen). As proteínas detectadas poderão servir como base de estudo na elaboração de métodos diagnósticos sensíveis e específicos, no desenvolvimento de vacinas, além de possibilitarem novos estudos para terapias mais eficazes.(AU)

Brazilian Spotted Fever (BSF) is an infectious disease transmitted by ticks to humans. A new human rickettsial infection was reported to cause spotted fever in the State of São Paulo and was named Rickettsia sp. Strain Atlantic Forest. This study aimed to detect and identify proteins with potential to stimulate the immune system of mammalian host of this new strain described. Therefore, we performed total protein extraction Rickettsia sp. Strain Atlantic Forest. The extracted proteins were fractionated by electrophoresis. The protein bands were transferred to nitrocellulose membrane by electrical migration and subjected to Western blot for protein detection. In all, seven immunoreactive proteins were detected. Two proteins showed higher abundance, with molecular weight of 200 and 130 kDa respectively. By comparing existing proteomic maps and the molecular weight of these proteins showed that, it is suggested that the two proteins detected representing rOmpA (200 kDa) and rOmpB (130 kDa). The other detected proteins had lower occurrence and molecular weight less than 78 kDa, which may represent members of the cell surface antigens Family (Sca - Surface cell antigen). The detected proteins may serve as a study based on the development of sensitive and specific diagnostic methods in the development of vaccines and they enable further studies to more effective therapies.(AU)

Fibrosis quística y tamizaje neonatal / Newborn screening and cystic fibrosis

Newborn screening for cystic fibrosis is a public health strategy that as been associated with early improved nutrition and survival, and the potential for preventing severe health problems. Various protocols have been employed, being the measurement of immunoreactive trypsynogen (IRT) in the first few days of life the first step in all of them. The second tier can include IRT/IRT, IRT/DNA, IRT/PAP (pancreatitis associated protein). Protocol selection depends on the priorities for each country but at present there is no optimal strategy.

El tamizaje neonatal para fibrosis quística es una estrategia de salud pública que ha demostrado beneficios nutricionales, aumento de la sobrevida y potencialmente prevención de problemas severos de salud. En el mundo se usan variados protocolos, sin embargo en todos el primer paso es la determinación de tripsinógeno inmunorreactivo (IRT) en sangre, tomado del talón del recién nacido. El segundo paso incluye la determinación de un segundo IRT o determinación de DNA o PAP (Proteína asociada a pancreatitis). La selección del protocolo a seguir depende de cada país pero hasta ahora no hay una estrategia óptima.

Analysis of relationship between stress hyperglycemia and variation of serum insulin in critical ill children / 中华实用儿科临床杂志

Objective To study the relationships between the level of blood glucose in critical ill children with the degree of critical illness and the variation of immunoreactive insulin (IRI) and true insulin (TI).Methods Fiftyeight children form the Neonatal Intensive Care Unit (PICU) and Department of Respiration were enrolled in this study.The children were divided into PICU group (42 cases) and control group (16 cases).The PICU group were scored pediatric critical score in 24 hours after admission.The 42 critical ill children were divided into stress hyperglycemia group (20 cases) and non-stress hyperglycemia group (22 cases) according to their blood glucose levels.The IRI,TI,C-Peptide and blood glucose were measured.Results The pediatric critical illness score of stress hyperglycemia group [(74.80 ± 8.07) scores] was significantly lower than that of non-stress hyperglycemia group [(84.36 ±9.46) scores] (t =1.964,P ＜ 0.05).The death rate of stress hyperglycemia group (45.0％,9/20 cases) was significantly higher than that of non-stress hyperglycemia group (13.6％,3/22 cases) (x2 =5.05,P ＜ 0.05).The IRI,TI and C-Peptide of stress hyperglycemia group were significantly higher than those of non-stress hyperglycemia group and control group(F =136.90,61.25,45.89,all P ＜ 0.05).The TI/IRI of stress hyperglycemia group was significantly lower than that of non-stress hyperglycemia group and control group (F =27.64,P ＜ 0.05).The TI,IRI and C-Peptide of stress hyperglycemia group were higher than after admission (t =2.241,2.087,2.014,all P ＜ 0.05).Conclusions The children with critical illness have stress hyperglycemia and the component of insulin is changed,and the absolute level as well as the rate of TI and TI/IRI are descended.

Quantification of argyrophillic, argentaffin and insulin immunoreactive cells in the small intestine in the opossum Didelphis aurita (Wied-Neuwied, 1826) / Quantificação das células argirófilas, argentafins e imunorreativas à insulina nos diferentes segmentos do intestino delgado do gambá Didelphis aurita (Wied-Neuwied, 1826)

The objective of this study was to quantify argyrophillic, argentaffin and insulin immunoreactive endocrine cells in the different segments of the small intestine of Didelphis aurita and measure probable differences in the number of these cells between adult and post-pubertal animals. Biological material consisted of ten male and female opossums specimen, divided in two groups according to weigh. The utilized staining techniques were Grimelius, modified Masson-Fontana and direct immunoperoxidase. Results indicated a predominance of argyrophillic cells in the small intestine of opossums from class 1 and 2, with an average of 52.58 and 56.15 cells mm-2, respectively; of which, the average number of total endocrine cells, argyrophillic and argentaffin cells decreased distally in the intestinal segments of opossums from classes 1 and 2. No significant difference was observed for the insulin immunoreactive cells between the intestinal segments of animals from class 2. A greater number of insulin immunoreactive cells was encountered in the jejunum and ileum of animals from class 2 when compared to the same segment in animals from class 1.

Os objetivos deste trabalho foram quantificar as células endócrinas argirófilas, argentafins e imunorreativas à insulina nos diferentes segmentos do intestino delgado de gambás Didelphis aurita e mensurar prováveis diferenças no número destas células entre animais adultos e pós-púberes. Dez exemplares de gambás D. aurita machos e fêmeas foram divididos em dois grupos de acordo com o peso. As técnicas de coloração utilizadas foram Grimelius, Masson-Fontana modificado e Imunoperoxidase direta. Os resultados indicaram um predomínio das células argirófilas no intestino delgado de gambás da classe 1 e 2, com uma média de 52,58 e 56,15 células mm-2, respectivamente. O número médio de células endócrinas totais, células argirófilas e argentafins decresceu distalmente nos segmentos intestinais dos gambás das classes 1 e 2. Nenhuma diferença significativa foi observada para as células imunorreativas à insulina entre os segmentos intestinais dos animais da classe 2. Foi encontrado maior número de células imunorreativas à insulina no jejuno e íleo de animais da classe 2 quando comparado ao mesmo segmento em animais da classe 1.

Quantification of argyrophillic, argentaffin and insulin immunoreactive cells in the small intestine in the opossum Didelphis aurita (Wied-Neuwied, 1826) - doi: 10.4025/actascibiolsci.v33i4.8644 / Quantification of argyrophillic, argentaffin and insulin immunoreactive cells in the small intestine in the opossum Didelphis aurita (Wied-Neuwied, 1826) - doi: 10.4025/actascibiolsci.v33i4.8644

The objective of this study was to quantify argyrophillic, argentaffin and insulin immunoreactive endocrine cells in the different segments of the small intestine of Didelphis aurita and measure probable differences in the number of these cells between adult and post-pubertal animals. Biological material consisted of ten male and female opossums specimen, divided in two groups according to weigh. The utilized staining techniques were Grimelius, modified Masson-Fontana and direct immunoperoxidase. Results indicated a predominance of argyrophillic cells in the small intestine of opossums from class 1 and 2, with an average of 52.58 and 56.15 cells mm-2, respectively; of which, the average number of total endocrine cells, argyrophillic and argentaffin cells decreased distally in the intestinal segments of opossums from classes 1 and 2. No significant difference was observed for the insulin immunoreactive cells between the intestinal segments of animals from class 2. A greater number of insulin immunoreactive cells was encountered in the jejunum and ileum of animals from class 2 when compared to the same segment in animals from class 1.

The objective of this study was to quantify argyrophillic, argentaffin and insulin immunoreactive endocrine cells in the different segments of the small intestine of Didelphis aurita and measure probable differences in the number of these cells between adult and post-pubertal animals. Biological material consisted of ten male and female opossums specimen, divided in two groups according to weigh. The utilized staining techniques were Grimelius, modified Masson-Fontana and direct immunoperoxidase. Results indicated a predominance of argyrophillic cells in the small intestine of opossums from class 1 and 2, with an average of 52.58 and 56.15 cells mm-2, respectively; of which, the average number of total endocrine cells, argyrophillic and argentaffin cells decreased distally in the intestinal segments of opossums from classes 1 and 2. No significant difference was observed for the insulin immunoreactive cells between the intestinal segments of animals from class 2. A greater number of insulin immunoreactive cells was encountered in the jejunum and ileum of animals from class 2 when compared to the same segment in animals from class 1.

Neonatal screening for cystic fibrosis in São Paulo State, Brazil: a pilot study

Cystic fibrosis is one of the most common autosomal recessive hereditary diseases in the Caucasian population, with an incidence of 1:2000 to 1:3500 liveborns. More than 1000 mutations have been described with the most common being F508del. It has a prevalence of 23-55 percent within the Brazilian population. The lack of population-based studies evaluating the incidence of cystic fibrosis in São Paulo State, Brazil, and an analysis concerning the costs of implantation of a screening program motivated the present study. A total of 60,000 dried blood samples from Guthrie cards obtained from April 2005 to January 2006 for neonatal screening at 4 reference centers in São Paulo State were analyzed. The immunoreactive trypsinogen (IRT)/IRT protocol was used with the cut-off value being 70 ng/mL. A total of 532 children (0.9 percent) showed IRT >70 ng/mL and a 2nd sample was collected from 418 (80.3 percent) of these patients. Four affected children were detected at two centers, corresponding to an incidence of 1:8403. The average age at diagnosis was 69 days, and 3 of the children already showed severe symptoms of the disease. The rate of false-positive results was 95.2 percent and the positive predictive value for the test was 8 percent. The cost of detecting an affected subject was approximately US$8,000.00 when this cystic fibrosis program was added to an existing neonatal screening program. The present study clearly shows the difficulties involved in cystic fibrosis screening using the IRT/IRT protocol, particularly in a population with no long-term tradition of neonatal screening.

Assays of immunoreactive insulin and specific insulin in evaluating islet function and insulin sensitivity / 中华内分泌代谢杂志

Comparison of assays of immunoreactive insulin (IRI) and specific insulin in evaluating islet β cell function and insulin sensitivity suggested that there were no significant differences in individuals with different glucose tolerance impairment by two assays. The evaluation of islet β cell function using IRI and insulin sensitivity is still valid in clinical practice.

Clinical significance of histological examination and noninvasive measurements in renal osteodystrophy / 中华肾脏病杂志

Objective To analyze the histological changes of bone diseases and to investigate the noninvasive measurements for diagnosing renal osteodystrophy (ROD) in maintenance dialysis patients . Methods Ninety-one patients were selected to receive bone biopsy . The bone samples were stained with HE, toluidine blue and Masson, and were examined with light microscopy . The levels of immunoreactive parathyroid hormone (iPTH), osteoprotegerin (OPG),sRANKL and osteocalcin (OCN) were determined in the patients enrolled from 2004 to 2006 . The level of iPTH was measured by radioimmunoassay . OPG and sRANKL were measured by ELISA,and OCN was measured by chemiluminescence . Results The incidence of ROD in the maintenance patients was 100% . According to the histological appearance, 50 cases (54 .9%) were high turnover bone disease (secondary hyperparathyroid bone disease), 9 cases (9 .9%) were low turnover bone diseases(osteomalacia and adynamic bone disease), and 32 cases(35 .2% ) were mixed bone disease . The level of iPTH in patients with ROD was significantly increased compared with healthy controls . It was the lowest in low turnover bone diseases . There was no difference among three types of ROD . OPG level was significantly increased compared with healthy controls [(2176 .58±1576 .08) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . The level in high turnover bone diseases was higher than that of the healthy controls [(2261 .85±1712 .22) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . There was no difference among three types of ROD .sRANKL level in high turnover bone disease was significantly increased compared with healthy controls [(0 .328±0 .524)pmol/L vs (0 .084±0 .190) pmol/L, P＜0 .05] . OCN level was also higher than that of the healthy controls (P＜0 .05), and the OCN level in low turnover ROD was the lowest among three types of ROD . OCN level in mixed ROD was dramatically increased as compared to low turnover ROD [(226 .633±66 .455) pmol/L vs (193 .03±104 .269) pmol/L, P ＜0 .05] .Conclusions The histological changes of bone disease can be indicated by iPTH level, but the types of ROD can not be distinguished according to iPTH level neither be differentiated by the levels of OPG, sRANKL and OCN . Bone histomorphometry is still the golden standard for diagnosing renal osteodystrophy .

Immunocytochemical study of the distribuition of endocrine cells in the pancreas of the Brazilian sparrow species Zonotrichia Capensis Subtorquata (Swaison, 1837)

In the present study, we investigated types of pancreatic endocrine cells and its respective peptides in the Brazilian sparrow species using immunocytochemistry. The use of polyclonal specific antisera for somatostatin, glucagon, avian pancreatic polypeptide (APP), YY polypeptide (PYY) and insulin, revealed a diversified distribution in the pancreas. All these types of immunoreactive cells were observed in the pancreas with different amounts. Insulin- Immunoreactive cells to (B cells) were most numerous, preferably occupying the central place in the pancreatic islets. Somatostatin, PPA, PYY and glucagon immunoreactive cells occurred in a lower frequency in the periphery of pancreatic islets.

Os tipos de células endócrinas e seus respectivos peptídeos reguladores foram estudados imunocitoquimicamente no pâncreas do tico-tico, espécie Zonotrichia capensis subtorquata, empregando-se o método imunocitoquímico ABC - Peroxidase (Complexo Avidina - Biotina - Peroxidase) e anti-soros específicos para somatostatina, ao glucagon, ao polipeptídeo pancreático aviário (PPA), ao polipeptídeo YY (PYY) e à insulina. Todos estes tipos de células imunorreativas foram observadas no pâncreas em quantidades diferentes. As células imunorreativas à insulina (células B) foram as mais numerosas, ocupando preferencialmente, a região central das ilhotas pancreáticas. As células endócrinas imunorreativas à somatostatina, PPA, PYY e glucagon localizaram-se predominantemente na periferia das ilhotas.

The Ultrastructure of TRPV1-positive Nerve Terminals in the Human Tooth Pulp / 대한해부학회지

The vanilloid receptor TRPV1 has been suggested to play an important role in thermal nociception and inflammatory hyperalgesia. In our previous study, we examined the expression of TRPV1 and colocalization of TRPV1 with substance P (SP) and calcitonin gene related peptide (CGRP) through fluorescence immunocytochemistry. Here, we investigated ultrastructural characteristics of TRPV1 immunoreactive fibers in the human tooth pulp through preembedding immunocytochemistry. TRPV1 immunoreactivity was present in the unmyelinated nerve fibers in the tooth pulp. There were two types of TRPV1 IR nerve fibers identified in the human tooth pulp: one containing clear round vesicles and many dense-cored vesicles, the other containing clear round vesicles and few dense-cored vesicles. TRPV1 immunoreactive fibers were constant in diameter without swellings along the length. Boutons en passant and boutons terminaux usually observed in the CNS were not observed in the TRPV1 immunoreactive fibers. Many vesicles were accumulated in the TRPV1 immunoreactive fibers, however synaptic structure was not found. It is known that dense-cored vesicles contain neuropeptides such as SP and CGRP and clear round vesicles contain neurotransmitter such as glutamate. Taken together, our results suggest that TRPV1 immunoreactive fibers showing distinct ultrastructructural features may be involved in inflammatory hyperalgesia and thermal nociception in the tooth pulp.

Immunohistochemical Study of the Endocrine Cells in the Pancreas of the Carp,Cyprinus carpio (Cyprinidae)

The regional distribution and relative frequency of some endocrine cells in the pancreas of the carp, Cyprinus carpio Linnaeus, belonging to the family Cyprinidae in the order Cypriniformes, were observed using specific mammalian antisera against insulin, glucagon, somatostatin and human pancreatic polypeptide (hPP) by peroxidase antiperoxidase (PAP) method. The pancreas was divided into four regions (principal and secondary islets, exocrine and pancreatic duct regions). In addition, the pancreatic islet regions were further subdivided into three regions (central, mantle and peripheral regions) and the pancreatic duct regions were subdivided into two regions (epithelial and subepithelial regions). Spherical to spindle or occasionally round to oval shaped immunoreactive (IR) cells were demonstrated in the pancreatic islets, exocrine and pancreatic duct. In the principal islet regions, some cells were also detected in the other regions, most of insulin- and somatostatin-IR cells were located in the central regions, and glucagon- and hPP-IR cells were situated in the peripheral regions. In this regions, insulin-IR cells were most predominant cell types and then, glucagon, somatostatin and hPP in that order. In the secondary islet regions, the regional distribution and relative frequency of these four types of endocrine cells were quite similar to those of the principal islets except for cell clusters consisted of hPP-IR cells that were situated in the peripheral to mantle regions. In the pancreatic duct regions, all four major pancreatic endocrine cells were demonstrated in the inter-epithelial cells and/or basal regions of the epithelial linning. In addition, cell clusters composed of numerous insulin-, moderate glucagon- and somatostatin-IR cells of low frequency were also observed in the subepithelial regions of the pancreatic duct. In the exocrine regions, insulin-, glucagon-, somatostatin- and hPP-IR cells were located in the inter-acinus regions with rare, a few, moderate and moderate frequencies, respectively. In conclusion, the regional distribution and relative frequency of four major pancreatic endocrine cells, insulin-, glucagon-, somatostatin- and hPP-IR cells, in the pancreas of the carp showed general patterns which were observed in other stomachless teleost. However, some species- dependent different distributional patterns and/or relative frequencies were also demonstrated.

First-phase insulin secretion and its components in the first-degree relatives of type 2 diabetics / 中华内分泌代谢杂志

Objective To investigate the changes of first-phase insulin secretion and its components in the first-degree relatives of Chinese type 2 diabetics with normal glucose tolerance and their correlation with type 2 diabetes. Methods The first-degree relatives of type 2 diabetes (Group B, n=35), newly diagnosed type 2 diabetic patients (Group C, n=35) and health subjects (Group A, n=21) were recruited. Immune reactive insulin (IRI), proinsulin (PI), and true insulin (TI) were determined during intravenous glucose tolerance test (IVGTT) in each subjects. IRI was determined with radioimmunoassay kit, while PI and TI with ELISA kits. Results (1) For the fasting sera, the levels of IRI and PI showed significant differences (both P

Increased levels of multiple forms of dihydrofolate reductase in peripheral blood leucocytes of cancer patients receiving haematopoietic colony-stimulating factors: interim analysis

The precise mechanism whereby granulocytes proliferate when haematopoietic colony stimulating factors (CSFs) are used in neutropenic cancer patients is poorly understood. The purpose of this study was to investigate whether these cytokines bring about leucocyte proliferation by increasing the levels of multiple forms of dihydrofolate reductase (DHFR). Blood samples were collected from 36 cancer patients (25 males and 11 females) with chemotherapy-induced neutropenia. One sample of blood from each patient was obtained before therapy either with CSF, such as granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) or with placebo, and another one at the time of resolution of neutropenia. Peripheral blood leucocytes in these blood samples were counted, separated and lysed. From lysates, cytoplasmic samples were prepared and analyzed for active DHFR by a methotrexate-binding assay and for total immunoreactive DHFR by an enzyme linked immunosorbent assay. The increase in total leucocyte count (TLC) was most prominent (P < 0.005) in the CSF group and less so (P < 0.05) in the placebo group. The mean +/- SD concentration values of active DHFR before and after stimulation with GM-CSF found were to be 0.34 +/- 0.4 ng/mg protein and 0.99 +/- 0.82 ng/mg protein, respectively, and in the group treated with G-CSF, 0.24 +/- 0.32 ng/mg protein and 1.18 +/- 2.4 ng/mg protein, respectively. This increase in active DHFR after stimulation with CSF was statistically significant (P <0.05). Similarly, concentration values of immunoreactive but nonfunctional form of DHFR (IRE) were 110 +/- 97 ng/mg protein and 605 +/- 475 ng/mg protein before and after stimulation with GM-CSF, and 115 +/- 165 ng/mg protein and 1,054 +/- 1,095 ng/ mg protein before and after stimulation with G-CSF. This increase in concentration of IRE after stimulation with GM-CSF or G-CSF was statistically significant (P < 0.005). In the control group, there was an increase in the concentration of both active DHFR and IRE after treatment with placebo. However, this was not statistically significant. Resolution of neutropenia was quicker in the groups treated with CSF compared to the control group. Results of this study indicate that colony stimulating factors (G-CSF and GM-CSF) induce white cell proliferation by increasing the levels of multiple forms of DHFR.

A Case of Digoxin-like Immunoreactive Substance(DLIS) Poisoning After Ingestion of Fried Water Toad (Bufo Stejnegeri Schmidt) / 대한내과학회지

Everybody knows that toads are not eatable because of their toxins. But water toads are very similar to frogs morphologically, so fried water toads are eaten by some people on occasion. It has been studied that the granular glands of the toads secrete toxins known as digoxin-like immunoreactive substances (DLIS) such as bufogenins and bufotoxins. DLIS have cross-reactivity with micro particle enzyme immunoassay for the quantitative mea surement of digoxin. We report a case of DLIS poisoning in 56-year-old male patient who showed vomiting, syn cope, and marked bradyarrhythmia with block (heart rate down to 29 beats per minute) after ingestion of the fried water toads (Bufo stejnegeri Schmidt).

Immunohistochemical Study on the Changes of Neuropeptide Y Immunoreactive Neurons in the Corpus Striatum and Motor System of Aged Rat / 대한해부학회지

Decreased number of the Neuropeptide-Y[NPY] immunoreactive neurons in the corpus striatum and primary motor cortex of aged rat was detected by the immunohistochemical method. The animals were categorized into control and aged group and we used 10 Sprague-Dawley rat weighing 250-300gm for control group. 10 Sprague-Dawley rat weighing over 600gm for aged group. The number of NPY-immunoreactive neurons in corpus striatum and primary motor cortex were counted under the light microscope and the following results were obtained. 1. The NPY-immunoreactive neurons were evenly distributed in corpus striatum and in the primaty motor cortex, the NPY-immunoreactive neurons were concentrated within the layer II, III and layer V, VI. The typical NPY-immunoreactive perikarya was multipolar shape. 2. Decreased number of NPY-immunoreactive neurons were detected in some areas of corpus striatum and primary mortor cortex of the aged rat. 3. Decrease of NPY-immunoreactive neurons were most prominent in the caudate-putamen and there were moderate decrease of NPY-immunoreactive neurons in the primary motor cortex, mild decrease of NPY-immunoreactive neurons in the nucleus accumbens but the NPY-immunoreactive neurons were not observed in the globus pallidus in both control and aged rat. NPY is supposed to act as a neurotransmitter of local circuit neurons in the striatum and may exert its potent vasoconstrictor effects on cerebral vessels which influences on the microcirculation of cerebral cortex and striatum. So our results seems to provide an important data on change of the function in the striatum and primary motor cortex of aged rat brain.