Comparasion on Effect of Xuefu Zhuyutang and Modified Erchentang on Nonalcoholic Fatty Liver in Mice / 中国实验方剂学杂志

Objective:To compare the effect and mechanisms of modified Erchentang and Xuefu Zhuyutang on high-fat diet-induced apolipoprotein-E knockout (ApoE-/-) mice nonalcoholic fatty liver disease (NAFLD). Method:Ten C57/BL6J mice were taken as normal control group and fed with normal feed. Totally 30 ApoE-/- mice were fed with high-fat diet to establish a disease model for 4 weeks. After 4 weeks, the 30 ApoE-/- mice were divided into model group, Xuefu Zhuyutang group (hereinafter referred to as Huoxue group) and modified Erchentang group (hereinafter referred to as Huatan group) by random number table method, with 10 in each group. The normal group and the model group were intragastrically administered with normal saline. The drug-administered group was intragastrically administered at a dosage that was ten times of the adult dose, once a day, for 8 weeks. Serum and liver were collected after the end of the 12-week experiment. The serum lipid and liver function levels of each group were measured, and the liver pathological morphology was observed. Protein and mRNA expressions of liver inflammatory mediators interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), monocyte chemotactic factor-1 (MCP-1) were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot. Result:The results of serum lipids and liver function showed that compared with the normal group, serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the model group were significantly increased, while serum high-density lipoprotein (HDL) was significantly reduced (P<0.01). Compared with the model group, serum TG ，LDL and ALT were significantly decreased, HDL was significantly increased in the Huoxue group (P<0.05). The serum levels of TC, TG， LDL, AST and ALT in the Huatan group were significantly decreased，HDL was significantly increased (P<0.05，P<0.01), and TG was decreased. The mice serum HDL in the Huatan group was higher than that in the Huoxue group. The serum ALT in the Huoxue group was lower than that in the Huatan group. The pathological observation showed that compared with the normal group, hepatocytes in the model group had severe steatosis with many lipid droplet vacuoles, suggesting that the mouse NAFLD model was successful. Compared with the model group, each administration group alleviated hepatocyte steatosis, with no significant difference between the two administration groups. Western blot and Real-time PCR results showed that compared with the normal group, protein and mRNA expression levels of TNF-α, IL-1β, MCP-1, and MMP-9 in the model group were significantly increased (P<0.05，P<0.01). Compared with the model group, the Huoxue group significantly down-regulated the expressions of IL-1β, MCP-1 protein and MCP-1 mRNA(P<0.05，P<0.01). The Huatan group significantly reduced the expressions of IL-1β, TNF-α, MMP-9, MCP-1 protein, TNF-α and MMP-9 mRNA(P<0.05，P<0.01). Conclusion:Modified Erchentang and Xuefu Zhuyutang can alleviate the therapeutic effect of NAFLD mice to a certain extent, modified Erchentang has a better therapeutic effect.

Effect of Mangiferin on Morphology of Relevant Vessels and MCP-1/CCR-2 Pathway in Spontaneously Hypertensive Rats / 中国实验方剂学杂志

Objective:To observe the morphological changes of carotid artery, thoracic aorta and superior mesenteric artery in spontaneously hypertensive rats(SHR), in order to further study the effect of Mangiferin on the expressions of inflammatory factors and monocyte chemoattract protein-1 (MCP-1)/c-chemokine receptor type 2 (CCR-2) pathway in SHR. Method:Forty spontaneously hypertensive rats were randomly divided into model group, benazepril group (10 mg·kg-1·d-1) and low, medium and high-dose mangiferin groups (25, 50, 100 mg·kg-1·d-1). Eight male WKY rats of the same age were selected as normal control group. Systolic blood pressure was observed every two weeks after eight weeks of administration. Morphology of carotid artery, thoracic aorta and superior mesenteric artery was observed by hematoxylin-eosin (HE) staining. Immunohistochemical assay (IHC) and Western blot were used to detect MCP-1 and CCR-2 protein expressions in thoracic aorta. MCP-1 and CCR-2 mRNA expression levels in thoracic aorta were detected by Real-time quantitative fluorescence PCR (Real-time PCR). Result:Compared with the normal group, the inflammatory cells in the model group increased significantly, the systolic blood pressure was significantly higher than that in the WKY group (PPPPConclusion:There are inflammation damages in carotid artery, thoracic aorta and superior mesenteric artery of spontaneously hypertensive rats. Mangiferin has an anti-inflammatory effect by possibly inhibiting the expressions of MCP-1/CCR-2 pathway in SHR vessels.

Mechanism of Modified Erchentang on Signaling Pathway of TLR4/MyD88/NF-κB in Lung Tissue of Rats with Chronic Obstructive Pulmonary Disease / 中国实验方剂学杂志

Objective: To study the effect of modified Erchentang on expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB (NF-κB) genes in the lung tissue homogenate of rats with chronic obstructive pulmonary disease (COPD). Method: Forty SD rats were randomly divided into normal group, model group, modified Erchentang group and EVP4593 (NF-κB inhibitor) group. Rat COPD models were prepared through cigarette smoke and tracheal dripping with lipopolysaccharide (LPS). After the modeling, normal and model groups were intragastrically given normal saline solution, EVP4593 group was given EVP4593(1 mg · kg-1) through subcutaneous injection, and modified Erchentang group was given corresponding herbal drugs intragastrically (10 g · kg-1) for 14 days. The levels of high mobility group box 1(HMGB1), chemokines CXCL-2, CXCL-3 and monocyte chemoattractant protein-1 (MCP-1) in rats serum were detected by enzyme-linked immunosorbent assay in rats serum. The expressions of Toll-like receptors 4(TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB p65 (NF-κB p65) mRNA were detected by Real-time fluorescence quantitative PCR (Real-time PCR) method. Western blot were used to detect the levels of TLR4, MyD88, NF-κB p65 and p-NF-κB p65 protein. Immunohistochemistry (IHC) method was used to detect the localization and expressions of TLR4, MyD88 and p-NF-κB p65 protein in the lung tissue. Result: The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 were increased significantly (PPκB p65 mRNA and protein were decreased significantly (PConclusion: Modified Erchentang may inhibit the inflammatory response of COPD effectively. The mechanism may be related to the inhibition of the expressions of the signal molecule genes involved in the TLR4/MyD88/NF-κB pathway and the reduction of the release of HMGB1, CXCL-2, CXCL-3 and MCP-1.

Spinal CCL2 Promotes Central Sensitization, Long-Term Potentiation, and Inflammatory Pain via CCR2: Further Insights into Molecular, Synaptic, and Cellular Mechanisms / 神经科学通报·英文版

Mounting evidence supports an important role of chemokines, produced by spinal cord astrocytes, in promoting central sensitization and chronic pain. In particular, CCL2 (C-C motif chemokine ligand 2) has been shown to enhance N-methyl-D-aspartate (NMDA)-induced currents in spinal outer lamina II (IIo) neurons. However, the exact molecular, synaptic, and cellular mechanisms by which CCL2 modulates central sensitization are still unclear. We found that spinal injection of the CCR2 antagonist RS504393 attenuated CCL2- and inflammation-induced hyperalgesia. Single-cell RT-PCR revealed CCR2 expression in excitatory vesicular glutamate transporter subtype 2-positive (VGLUT2) neurons. CCL2 increased NMDA-induced currents in CCR2/VGLUT2 neurons in lamina IIo; it also enhanced the synaptic NMDA currents evoked by dorsal root stimulation; and furthermore, it increased the total and synaptic NMDA currents in somatostatin-expressing excitatory neurons. Finally, intrathecal RS504393 reversed the long-term potentiation evoked in the spinal cord by C-fiber stimulation. Our findings suggest that CCL2 directly modulates synaptic plasticity in CCR2-expressing excitatory neurons in spinal lamina IIo, and this underlies the generation of central sensitization in pathological pain.

Relation of Toll-like Receptor 4 and Monocyte Chemoattractant Protein-1 with Acute Coronary Syndrome / 现代检验医学杂志

Objective To investigate the relation of Toll-like receptor 4 (TLR4)and monocyte chemoattractant protein-1 (MCP-1)with acute coronary syndrome.Methods 60 patients with coronary heart disease patients including 39 cases of a-cute coronary syndrome (ACS group),21 cases of stable angina (SA group)from January 2015-March 2016 in Xi’an First Hospital were in this observation.ACS group was divided into acute myocardial infarction (AMI,n=21)and unstable angina pectoris (UAP,n=18).Another 30 cases with angiographically normal was included in control group.Enzyme-linked immu-nosorbent assay (ELISA)detection was used to detect the serum content of MCP-1,TLR4 expression was tested by flow cy-tometry.Triglyceride (TG),total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C)and high-density lipopro-tein cholesterol (HDL-C)were detected.Results TLR4 levels of ACS group was significantly higher than SA group (t=4.455,P=0.021).Serum MCP-1 levels of ACS group were significantly higher than SA group (t=3.220,P=0.002)and control group (t=6.197,P=0.000).MCP-1 levels of ACS group were higher than control group (t=2.306,P=0.025). The Spearman correlation test TLR4 was positively with MCP-1 (r=2.389,P=0.025).Conclusion MCP-1 and TLR4 may determine the severity of coronary heart disease,combined detection of MCP-1 and TRL4 can provide the clinical value for early diagnosis and treatment of ACS.

Effects of Lipoic Acid on Proteinuria,Serum and Urine MCP -1 Secretion in Patients with Stage IV Type 2 Diabetic Nephropathy / 医学研究杂志

Objective To investigate the effect of on serum and urine MCP -1 secretion in patients with Stage IV Type 2 diabetic nephropathy treated by lipoic acid .Methods We enrolled 76 diabetic nephropathy patients ,who were randomly divided into two groups . Patients in group T (24 males and 14 females) were treated by lipoic acid 0.6 gram per day.Those in group C (22 males and 16 fe-males) were treated by routine drugs.The serum creatinine (Scr), urea nitrogen (BUN), fasting blood sugar(FBS) and 24h urinary pro-tein,serum and urine MCP -1 secretion were measured at the experiment onset and 3 weeks later.Results There was no significant difference in the serum creatinine , urea nitrogen, fasting blood sugar, HbA1c between the two groups neither at the experiment onset nor after 3 weeks.Compared to experiment onset , 24h urine protein (Tp/24h), serum and urine MCP-1 secretion were all significantly de-creased (P<0.05) in group T after 3 weeks.Compared to group C (2.41 ±0.91g/24h, 91.45 ±33.41pg/ml, 114.78 ±36.35pg/ml), all the levers in group T (1.89 ±0.72g/24h, 39.50 ±13.68pg/ml, 63.41 ±19.57pg/ml) was significantly decreased (P<0.05) after 3 weeks.There was a positive correlation between the serum , urine MCP-1 levels and Tp/24h (r=0.572, P<0.05;r=0.697,P<0.05).Conclusion Lipoic acid can reduce urine protein excretion in diabetic nephropathy patients , maybe by decreasing serum and u-rine MCP-1 secretion .

Perfis de compostos voláteis de banana submetidos a diferentes tratamentos pós-colheita e suas correlações com a expressão diferencial dos receptores de etileno / Volatile compounds profile of bananas submitted to different post-harvest treatments and its correlations to differential expression of ethylene receptors

O aroma de frutos é um atributo fortemente associado à qualidade, e quaisquer alterações ambientais ou tratamentos pós-colheita podem alterar a sua composição. Acredita-se que a biossíntese de voláteis seja um dos processos regulados pelo etileno. Estudos indicam que a expressão diferencial dos elementos que compõem os receptores de etileno desempenha importante papel na sinalização dos processos ligados ao amadurecimento, entre eles a formação do aroma. Os objetivos deste trabalho são: caracterizar as alterações decorrentes de tratamentos pós-colheita no aroma de banana durante o amadurecimento, e correlacionar com as variações nos padrões de expressão gênica diferencial dos receptores de etileno. Bananas pré-climatéricas variedade Nanicão foram divididas em quatro grupos: controle (não tratado), etileno (100ppm/12h), 1-MCP (100ppb/12h), armazenados a 20°C, e grupo frio (armazenado por 15 dias a 13°C). Foram analisados diariamente a produção de etileno e de CO2 por CG. Foram analisadas a cor da casca, açúcares solúveis e amido. Os compostos voláteis foram isolados por microextração em fase sólida (SPME) em frutos inteiros e polpas e analisados em CG-MS. Para confirmar os resultados e verificar se as alterações encontradas se repetem em outras variedades de bananas, o estudo foi repetido no Horticultural Sciences Department, na Universidade da Florida (EUA), em bananas var. 'Grand Naine'. Em paralelo, realizou-se a quantificação relativa da expressão dos receptores de etileno (ETR1, ERS1, ERS2 e ERS3) por PCR em tempo real. Com relação ao perfil de voláteis, os resultados indicam que os frutos não se diferenciam no período pré-climatérico. Porém, o perfil de voláteis do grupo controle foi significativamente diferente do grupo frio, tanto na polpa quanto no fruto inteiro no período climatérico. Esse efeito foi mais pronunciado na Nanicão do que na 'Grand Naine'. Compostos típicos como o acetato de isoamila foram drasticamente reduzidos nos frutos submetidos ao frio, e não foram encontrados na Nanicão. Não houve diferenças significativas com relação ao perfil de aromas entre o grupo controle e o grupo etileno. Com relação aos frutos tratados com 1-MCP observou-se o atraso na formação de alguns compostos sem alterar, contudo, o perfil final de voláteis. Com relação ao padrão de transcrição dos receptores de etileno, o frio reduziu o acúmulo dos transcritos do ETR1, ERS2 e ERS3 em todos os pontos. ERS1 parece estar correlacionado com a síntese de esteres. Os resultados sugerem que o mecanismo pelo qual o etileno regula o metabolismo de biossíntese de aromas parece contar com a participação relevante de determinados tipos de receptores. A correlação temporal encontrada entre as alterações no perfil de transcritos de três destes e os efeitos sobre a produção de compostos voláteis reforçam esta hipótese

Fruit aroma is an attribute strongly associated to quality, and any change in the environment or post-harvest treatment could affect its composition. Volatile biosynthesis is a process that is believed to be regulated by ethylene. Studies demonstrate that differential expression of ethylene receptors have an important role in fruit ripening processes, including aroma synthesis. The aims of this study are: evaluate modifications due to post-harvest treatment on the aroma of banana fruit during ripening, and correlate to variations on differential expression of ethylene receptors. Pre climacteric bananas of 'Nanicão' variety were divides in four groups: control (without treatment), ethylene (100ppm/12h), 1-MCP (100ppb/12h), stored at 20°C, and cold storage group (stored for 15 days at 13°C). Daily measurements were conducted of ethylene production and respiration using GC. Peel color, soluble sugars and starch were analyzed. Volatile compounds were isolated by solid phase microextraction (SPME) in whole fruits and pulp and analyzed by GCMS. To confirm the results ant to verify if the findings repeat in another banana variety, this study was repeated at Horticultural Sciences Department, at University of Florida (EUA), under supervision of Dr. Jeffrey K. Brecht, in bananas Cavendish cv. 'Grand Naine'. Also, relative quantification of the expression of ethylene receptors (ETR1, ERS1, ERS2 and ERS3) was analyzed using real time PCR. Regarding the volatile profile, groups did not differentiated in pre-climacteric period. But the volatile profile of control group significantly differentiates from cold storage group, in both pulp and whole fruit, in post climacteric period. This effect was more pronounced in bananas 'Nanicão' than 'Grand Naine'. Typical banana aroma compounds like isoamyl acetate were drastically reduced in fruits under cold storage, and were not found in 'Nanicão'. There were not any significant differences between control group and ethylene treated. Regarding 1-MCP treated fruits, there was a delay on the synthesis of some compounds without affecting the final volatile profile. Regarding the transcription pattern of ethylene receptors, cold storage reduced mRNA of ETR1, ERS2 and ERS3 in all samples. ERS1 receptor seems to be correlated to ester synthesis. These results suggest that the mechanism whereby the ethylene regulates the biosynthesis of aroma, seems to count with relevant participation of some receptors. The temporal correlation found in the differential expression of three receptors and the effect on volatile compounds synthesis reinforces this hypothesis

study of C reactive protein up-regulating monocyte chemotactic protein-1 expression in Metabolic Syndrome / 中国医师杂志

Objective To investigate the relationship between high sensitive C reactive protein(hsCRP) and monocyte chemotactic protein-1 (MCP-1) in patients with metabolic syndrome(MS).Methods Sixty-two patients with MS and 52 normal healthy individuals (NS) were recruited in the study.And the levels of hs-CRP and MCP-1 were detected by ELISA.The severity of Insulin resistance was calculated by homeostasismodel assessment of insulin resistance(HOMA-IR).The monocytes in blood were isolated,cultured and stimulated by different concentrations of CRP.MCP-lexpression of monocytes was measured by real-time PCR.Results Patients with MS displayed significantly higher serum levels of hs-CRP and MCP-1 compared with healthy control group [(80.26 ±35.52) nmol/L vs (3.12 ±2.55) nmol/L for hs-CRP,P ＜0.01 ; (15.35 ± 10.12) nmol/L vs (9.76 ± 6.15) nmol/L for MCP-1,P ＜ 0.05].There was significantly positive correlation between hs-CRP and MCP-1.Serum hs-CRP,MCP-1and insulin resistance index were positively correlated,respectively.In vitro experiments indicated that CRP could up-regulate MCP-1 expression of monocytes in a concentration-dependent way.When the concentration of CRP was 176nmol/L,the expression of MCP-1 could reach 105％ compared with non-stimulated group.Conclusion With the severity of the inflammatory response and enhanced insulin resistance,elevated serum CRP level may result in MCP-1 expression in patients with metabolic syndrome.

Firmeza de mangas 'Palmer' tratadas com 1-metilciclopropeno e armazenadas sob refrigeração / Firmness of mangos 'Palmer' treated with 1-methylcyclopropene and stored under refrigeration

O objetivo do trabalho foi avaliar o efeito do 1-MCP na firmeza de mangas ´Palmer', armazenadas sob refrigeração. As mangas foram tratadas com 1-MCP nas concentrações de 0, 100 e 150nL.L-1, durante 12 horas à 22ºC. Em seguida, foram armazenadas em câmara fria a 10 ± 1ºC e 80 a 90 por cento de UR, por um período de 35 dias. A cada 7 dias foram retirados 6 frutos de cada tratamento para as seguintes avaliações: firmeza, pectina total, percentagem de solubilização de pectinas, atividade da pectinametilesterase, poligalacturonase e â-D-galactosidase. O tratamento com 1-MCP, associado à refrigeração mostrou ser eficiente em prolongar a vida útil dos frutos. Os frutos tratados com 150nL.L-1 apresentaram menor percentagem de solubilização de pectina e mais firmes no final do armazenamento, quando comparados aos outros tratamentos. Após 35 dias de armazenamento refrigerado, todos os frutos estavam aptos para o consumo, indicando que outros estudos deverão ser realizados aumentando o tempo de armazenamento.

The objective of this work was to evaluate the effect of the 1-MCP in the firmness of mangos ´Palmer' stored under refrigeration. The mangos were treated with 1-MCP in concentrations of 0, 100 and 150nL.L-1, during 12 hours at the 22ºC. Soon after, they were stored in a cold chamber (10 ± 1ºC and 80 to 90 percent of UR), for a period of 35 days. Every 7 days were retired 6 fruits of each treatment for to the following evaluations: firmness, total pectin, percentage of solubilization of pectins, activity of the pectinmethylesterase, polygalacturonase and â-D-galactosidase activities. The treatment with 1-MCP, associated to the refrigeration showed to be efficient in prolonging the useful life of the fruits. The treated fruits with 150nL.L-1 presented smaller percentage of pectin solubilization and firmness in the end of the storage, when compared to the other treatments. After 35 days of refrigerated storage, all the fruits were capable for the consumption, indicating that other studies should be accomplished increasing the time of storage.

Inhibitory effect of rapamycin on human renal tubular epithelial-myofibroblast transition / 基础医学与临床

Objective Rapamycin (RAPA) is an anti-proliferative immunosuppressant and has been used to suppress rejection of transplanted organs. In present study, we observed the effect of rapamycin on epithelial-myofibroblast transition (EMT)of cultured HKC cells in vitro. Methods Cultured human proximal tubular epithelial cells (HKCs) were divided into three groups: blank control, treated with TGF-?1 (1 ?g/L) and treated with TGF-?1 (1 ?g/L) plus rapamycin (0.1, 1, 10, 100 ?g/L). The protein and mRNA for ?-SMA and E-cadherin in HKC cells were determined by Western Blot and RT-PCR.The mRNA level of Snail in HKC was detected by RT-PCR. Results Rapamycin dramatically abrogated TGF-?1 induced ?-SMA expression and restored E-cadherin expressionin HKC cells in a dose-dependent manner. At a concentration of 100 ?g/L, rapamycin almost completely blocked ?-SMA mRNA and protein expression induced by TGF-?1(1 ?g/L). Rapamycin also suppressed expression of ?-SMA in HKC cells at both mRNA and protein level in a time dependent manner.We also found rapamycin dramatically abrogated TGF-?1 induced Snail mRNA expression in HKC cells in a dose-dependent manner. Conclusion Rapamycin may inhibit EMT of tubular cells in vitro. The downregulation of Snail expression might be one of the mechanisms of rapamycin blocking EMT.

Expression of monocyte chemoattractant protein-1 in lupus nephritis and its clinical significance / 临床检验杂志

Objective To investigate the changes of monocyte chemoattractant protein-1(MCP-1)in serum and urine of lupus nephritis(LN)patients in active phase and remission phase.Methods The levels of MCP-1 in serum and urine of 58 LN patients(27 of active phase and 31 of remission phase)were measured by ELISA.The correlation between the levels of MCP-1 in variant phase of LN and other relevant factors were analyzed.Results The MCP-1 levels in sera of both active phase and remission phase of LN patients were markedly higher than those in controls(548.5?347.2 ng/L and 469.1?298.4 ng/L vs 273.3?146.7 ng/L,P0.05).Conclusion The MCP-1 levels in urine of LN patients is more suitable to evaluate the activity of disease as a sensitive marker.

Effect of telmisartan on plasma levels of inflammatory cytokine in unstable angina patients complicated with diabetes mellitus and hypertensive after percutaneous coronary intervention / 中国康复理论与实践

@# ObjectiveTo observe the effect of telmisartan on plasma levels of inflammatory cytokine in coronary artery disease (CAD) patients complicated with diabetes mellitus and hypertension after percutaneous coronary intervention.MethodsFifty CAD patients who just had undertook angioplasty and implanted stents were randomly divided into two groups, the test group (telmisartan group, n=25) and control group (perindopril group, n=25). After treatment, patients were followed-up for 6 months; plasma samples were collected from each patient before and after percutaneous coronary intervention. Then plasma levels of C-reactive protein (CRP), monocyte chemoattractant protein-1 (MCP-1) were measured by enzyme-linked immunoassay. The changes of cholesterol, fasting plasma glucose (FPG), insulin and insulin resistance index (IRI) were observed.ResultsAt the end of 6 months, plasma levels of CRP and MCP-1 of patients in the test group significantly declined (P<0.01), and showing a inversely correlation with FPG (P<0.01), and FPG, insulin and IRI also declined. In the control group, only CRP and MCP-1 declined (P<0.05). Meanwhile, the frequency of cardiovascular events in test group was significantly lower than that in the control group.ConclusionTelmisartan can decline plasma levels of CRP, MCP-1 and frequency of cardiovascular events as well as increasing insulin sensitivity and improving glucose metabolism to unstable angina patients.

Genetic Susceptibilities of MCP-1 and CCR2 to the Risk for Korean Allergic Rhinitis Patients Using Polymorphism Study / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: A biallelic A/G polymorphism in the Monocyte chemotactic protein (MCP) -1 at position -2518 has been found to affect the level of MCP-1 expression. To investigate if these polymorphisms in chemokine ligand and receptor genes are relevant for the development of allergic rhinitis, we investigated polymorphisms of MCP-1 and CC chemokine receptor 2 (CCR2) known as the receptor of MCP-1. MATERIALS AND METHOD: Blood samples for genetic analysis were obtained from 198 individuals with allergic rhinitis and from 278 healthy subjects without atopic diseases. Polymerase chain reaction-based assay for MCP-1 -2518 A/G (A/G polymorphism in the MCP-1 at position -2518) and CCR2 V64I polymorphisms (replacement of valine by isoleucine in CCR2 64) was used for genotyping. RESULTS: There were no differences in the frequencies of the genotypes in the controls and patients (p>0.05). The frequencies of the MCP-1 G and CCR2 A alleles were not statistically different between controls and allergic rhinitis patients (p>0.05). The odds ratios (95% confidence interval) of MCP-1 G/G and CCR2 A/A genotypes for allergic rhinitis were not statistically significant, whereas, alleles frequencies of MCP-1 -2518G and CCR2 A of controls were various according to the ethnic background. CONCLUSION: Our result suggests MCP-1 -2518 A/G and CCR2 V64I polymorphisms are not part of the factors contributing to genetical susceptibility in the development of allergic rhinitis in Koreans.

Elevated monocyte chemoattractant protein-1 in patients with Behcet's disease / 대한내과학회지

BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) belongs to C-C subfamily of chemokines, which stimulates the migration of monocytes. MCP-1 exerts various effects on the monocytes, including the induction of integrin and tissue factor, and synthesis of proinflammatory cytokines and arachidonic acid. In this study, we measured the MCP-1 levels in patients with Behcet's disease and evaluated the associations between the levels of MCP-1 and the level of other chemokines and various clinical features of Behcet's disease. METHODS: Serum samples were obtained from 67 patients with Behcet's disease and 30 healthy controls. Simultaneously, whole blood was isolated from patients (n=25) with Behcet's disease and healthy controls (n=11) and cultured in 24 well plates for 48 hours in the absence or presence of lipopolysaccharide (LPS) 5 microgram/mL, phytohaemagglutinin (PHA) 5 microgram/mL, phorbol 12-myristate 13-acetate (PMA) 50 ng/mL + ionomycin 5 microgram/mL. The MCP-1 concentrations were measured in the sera and culture supernatants by enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of serum MCP-1 were 2.5 times higher in patients with Behcet's disease than healthy controls. The patients with Behcet's disease had also higher levels of MCP-1 in the culture supernatants of whole blood cells, stimulated with LPS, but not with either PHA or PMA plus ionomycin, compared to healthy controls. Serum MCP-1 levels (n=67) were strongly correlated with serum RANTES, MIP-1alpha, IL-8 levels in Behcet's disease. In addition, the production of MCP-1 by whole blood culture from Behcet's disease patients (n=25) were also correlated well with those of RANTES, MIP-1alpha, and IL-8, when stimulated with LPS. However, MCP-1 levels in the sera and culture supernatants did not show any association with various clinical features of Behcet's disease including oral ulcer, genital ulcer, erythema nodosum, arthritis, uveitis, intestinal involvement, central nervous system involvement, and vascular thrombosis. CONCLUSION: In the sera and culture supernatants of whole blood stimulated with LPS, MCP-1 levels were higher in patients with Behcet's disease than controls and correlated well with RANTES, MIP-1alpha, IL-8 levels. These results suggest that the activation and migration of monocytes triggered by the increased production of MCP-1 may play a role in the pathogenesis of Behcet's disease.

Association of Amniotic Fluid Concentrations of Monocyte Chemotactic Protein-1 with Intrauterine Infections and Perinatal Outcomes in Preterm Labor / 대한산부인과학회잡지

OBJECTIVE: To examine if amniotic fluid (AF) monocyte chemotactic protein-1 (MCP-1) concentrations are useful in the identification of intrauterine infection and pregnancy outcomes in preterm labor with intact membranes. METHODS: The study population consists of 65 patients who received amniocentesis for preterm labor with intact membranes. Amniotic fluid was cultured for aerobic and anaerobic bacteria, as well as mycoplasmas. MCP-1 was determined by a sensitive and specific immunoassay. Fisher's exact test, Mann-Whitney U test, receiver operating characteristic curve, survival techniques, logistic regression, and Spearman correlation were used for statistical analysis. RESULTS: (1) Patients with a positive amniotic fluid culture had a significantly higher median AF MCP-1 concentration than those with negative results (median, 9.0 ng/mL; range, 0.45-40.5 ng/mL; vs median, 0.82 ng/mL; range, 0.06-30.1 ng/mL; P1.9 ng/mL had a significantly shorter median interval to delivery, the higher rate of histologic chorioamnionitis, preterm delivery within 2 and 5 days, and the occurrence of congenital proven or suspected sepsis than did those with AF MCP-1 concentration of <1.9 ng/mL after adjustment for gestational age (P<.05). (3) There was strong correlation between AF MCP-1 concentrations and AF interleukin-6 concentrations (r=.881, P<.001). CONCLUSION: AF MCP-1 determinations are useful in the identification of intrauterine infection, preterm delivery, and neonatal infectious complication in preterm labor with intact membranes.

Optimizing expression of recombinant human monocyte chemoattractant protein-1 in E.coli / 第二军医大学学报

Objective: To optimize the expression of recombinant human monocyte chemoattractant protein (rhuMCP-1) in E.coli DE3. Methods:With NBS-MICROS 15 L T.DR fermentor, pGEX-IN/huMCP-1 was constructed by our laboratory. Four parameters including pH,temperature,agitation rate and concentration of IPTG were studied by orthogonal experimental design. Results: It was found that the expression level was greatly affected by the amount of dissolved oxygen. This indicated that the agitation rate and ventilation amount were the most important parameters during fermentation. Examined by SDS-PAGE and gel scanning, the expression level of total protein was over 40% when agitation rate was 300 r/min and ventilation amount was 10 L/min. Conclusion: A method for high-level expression of huMCP-1 on pilot-scale is established, and it will be useful for large-scale industrial production of target protein.

Protective role of mycophenolate mofetil on renal tubulointerstitial injury of diabetic rats / 中国药理学通报

Aim To observe the effect of mycophenolate mofetil(MMF) on expressions of MCP-1 and CD68 in renal tubulointerstitial injury of diabetic rats and explore the mechanism of MMF′s protective role.Methods Diabetes was induced in uninephrectomized male Wistar rats by peritoneal injection of STZ (65 mg?kg-1). Rats were randomly divided into three groups:control group (NC),diabetic group (DM) and treated group (DM+MMF) with MMF(15 mg?kg-1?d-1).This study lasted for 8 weeks. 24 h urinary protein,blood glucose and the ratio of left kidney weight/body weight were determined after 8 weeks.The renal tubulointerstitial morphological change was observed,immunohistochemical method was used to analyze expressions of MCP-1 protein and CD68. Expression of MCP-1 mRNA in renal tissue was measured by quantitative Real-time PCR.Results Compared with NC group, serum glucose level,24 hour urinary protein and the ratio of left kidney weight/body weight were significantly increased(P

RELATIONSHIP OF INCREASED INTRACELLULAR S-ADENOSYLHOMOCYSTEINE WITH MONOCYTE CHEMOATTRACTANT PROTEIN -1 EXPRESSION IN HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS / 营养学报

Objective To explore the molecular basis of vascular endothelial cell's injury by cellular S-adenosylhomocysteine (SAH) accumulation, and the mechanism for cardiovascular disease induced by dietary high methionine intake. Method After human umbilical vein endothelial cells (HUVEC) were treated without (normal) or with different concentrations of potent S-adenosylhomocysteine hydrolase (SAHH) inhibitor 3-deazaadenosine (DZA) for 24, 48 or 72h, the level of intracellular inflammatory monocyte chemoattractant protein-1 (MCP-1) was measured with enzyme linked immunosorbent assay (ELISA). Then, the relative expression of MCP-1 mRNA was detected with quantitative reverse transcription polymerase chain reaction (qRT-PCR). Furtheremore, the methylated state of MCP-1 gene promoter was analyzed with methylation special PCR (MSP) method. Results The level of intracellular MCP-1 was increased in HUVEC incubated with DZA than normal (P