Role of NOD2 in dorsal root ganglion in development of neuropathic pain in rats / 中华麻醉学杂志

Objective:To evaluate the role of nucleotide-binding oligomerization domain-2 (NOD2) in dorsal root ganglion in the development of neuropathic pain (NP) in rats.Methods:Thirty-two adult male SPF Sprague-Dawley rats, weighing 240-260 g, aged 2-3 months, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), NP group (group S), negative control siRAN group (group N), and NOD2-siRNA group (group R). In N and R groups, 1×10 8 IFU/ml negative control siRNA and NOD2-siRNA 10 μl were intrathecally injected, respectively, once a day for 3 consecutive days.Normal saline 10 μl was intrathecally injected once a day for 3 consecutive days in C and S groups.The model of NP was established using spared nerve injury (SNI) at 2 weeks after intrathecal injection.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before surgery and 1, 3, 7, 10, 14 and 28 days after SNI.Animals were sacrificed after measuring pain threshold on day 28, and the dorsal root ganglions (DRGs) of the lumbar segment (L 4-6) were removed for determination of the expression of NOD2 (by Western blot) and expression of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-6 and NOD2 mRNA (using quantitative real-time polymerase chain reaction). Results:Compared with group C, MWT was significantly decreased at each time point after SNI, and the expression of NOD2 protein and mRNA and TNF-α, IL-1β and IL-6 mRNA in DRGs was up-regulated in group NP ( P<0.01). Compared with group NP, MWT was significantly increased at each time point after SNI, and the expression of NOD2 protein and mRNA and TNF-α, IL-1β and IL-6 mRNA in DRGs was down-regulated in group R ( P<0.01), and no significant change was found in the parameters mentioned above in group N ( P>0.05). Conclusion:The mechanism underlying the development of NP may be related to the up-regulation of NOD2 expression in DRGs, thus further promoting the expression of pro-inflammatory factors in rats.

Xenobiotic receptors in mediating the effect of sepsis on drug metabolism

Sepsis is an infection-induced systemic inflammatory syndrome. The immune response in sepsis is characterized by the activation of both proinflammatory and anti-inflammatory pathways. When sepsis occurs, the expression and activity of many inflammatory cytokines are markedly affected. Xenobiotic receptors are chemical-sensing transcription factors that play essential roles in the transcriptional regulation of drug-metabolizing enzymes (DMEs). Xenobiotic receptors mediate the functional crosstalk between sepsis and drug metabolism because the inflammatory cytokines released during sepsis can affect the expression and activity of xenobiotic receptors and thus impact the expression and activity of DMEs. Xenobiotic receptors in turn may affect the clinical outcomes of sepsis. This review focuses on the sepsis-induced inflammatory response and xenobiotic receptors such as pregnane X receptor (PXR), aryl hydrocarbon receptor (AHR), glucocorticoid receptor (GR), and constitutive androstane receptor (CAR), DMEs such as CYP1A, CYP2B6, CYP2C9, and CYP3A4, and drug transporters such as p-glycoprotein (P-gp), and multidrug resistance-associated protein (MRPs) that are affected by sepsis. Understanding the xenobiotic receptor-mediated effect of sepsis on drug metabolism will help to improve the safe use of drugs in sepsis patients and the development of new xenobiotic receptor-based therapeutic strategies for sepsis.

Effect of Signal-transducing Adaptor Protein 2 and Importin 5 Expression on Skeletal Muscle Protein Turnover / 中国康复理论与实践

@# Objective To explore the expression of signal-transducing adaptor protein 2 (STAP2) and importin 5 (Ipo5), and the effect of them on skeletal muscle protein turnover. Methods Twelve male Sprague-Dawley rats were divided into control group (n=6) and model group (n=6). The model group established skeletal muscle hypertrophy induced by blood flow restriction. The soleus muscle cells in both groups were cultured in vitro, and those in the model group infected with shRNA lentivirus to make the expression of STAP2 and Ipo5 silencing, over-expression and gene function salvage finally. The expression of STAP2, Ipo5 and myostatin were determined with Western blotting in soleus muscle in both groups.Results The expression of STAP2 increased (t=-11.786, P<0.001), while the expression of Ipo5 and myostatin decreased (t>14.039, P<0.001) in the model groups compared with those in the control group. Under the control group and the model group, and target genes silencing group, overexpressing group and rescuing group, the expression of myostatin increased with STAP2 decrease and Ipo5 increase.Conclusion The expression of STAP2 increasing and Ipo5 decreasing would promote the protein synthesis.

Role of NOD2-regulated Snail expression in epithelial-mesenchymal transition in podocyte of diabetic nephropathy / 中华肾脏病杂志

Objective To observe the expression of NOD2 and epithelial-mesenchymal transition (EMT) related proteins in podocytes in high glucose environment,and explore the molecular mechanism of NOD2 involved in EMT.Methods The human glomerular podocytes were the subjects of study.α-SMA and Nephrin expressions were detected by immunofluorescence;the mRNA and protein expressions of NOD2,Snail and EMT related proteins (α-SMA,Desmin,E-cadherin,Nephrin) were detected by real-time fluorescence quantitative PCR and Western blotting.The podocytes were stimulated by high-glucose after shRNA interfering the of NOD2 expression,and the expressions of Snail and subsequent EMT-related proteins were detected by Western blotting.Prior to the activation of NOD2 by muramyl dipeptide (MDP),shRNA was used to interfere with the expression of Snail.E-cadherin,Nephrin,Desmin,and α-SMA were detected by Western blotting.Results After 24 hours of high glucose stimulation,PCR and Western blotting results showed that the expressions of NOD2 and Snail were significantly increased;the expressions of epithelial phenotype proteins E-cadherin and Nephrin were down-regulated;and the expressions of interstitial phenotype proteins Desmin and α-SMA were increased (all P ＜ 0.05);while there was no significant change in the hypertonic control group.After interference with NOD2,the abnormal expression of Snail and EMT related proteins were all recovered.After interference with Snail expression,Compared with the MDP group,the protein expressions of E-cadherin and Nephrin were significantly increased (all P ＜ 0.05);the expressions of Desmin and α-SMA were significantly decreased.Conclusions High glucose can induce NOD2 expression in podocytes,and promote podocyte epithelial-mesenchymal transition by upregulating Snail expression.Gene intervention targeting the NOD2/Snail/EMT pathway can reduce high-glucose-induced podocyte injury and may provide new ideas for the treatment of diabetic nephropathy.

Insulin Substrate Receptor (IRS) proteins in normal and malignant hematopoiesis

The insulin receptor substrate (IRS) proteins are a family of cytoplasmic proteins that integrate and coordinate the transmission of signals from the extracellular to the intracellular environment via transmembrane receptors, thus regulating cell growth, metabolism, survival and proliferation. The PI3K/AKT/mTOR and MAPK signaling pathways are the best-characterized downstream signaling pathways activated by IRS signaling (canonical pathways). However, novel signaling axes involving IRS proteins (noncanonical pathways) have recently been identified in solid tumor and hematologic neoplasm models. Insulin receptor substrate-1 (IRS1) and insulin receptor substrate-2 (IRS2) are the best-characterized IRS proteins in hematologic-related processes. IRS2 binds to important cellular receptors involved in normal hematopoiesis (EPOR, MPL and IGF1R). Moreover, the identification of IRS1/ABL1 and IRS2/JAK2V617F interactions and their functional consequences has opened a new frontier for investigating the roles of the IRS protein family in malignant hematopoiesis. Insulin receptor substrate-4 (IRS4) is absent in normal hematopoietic tissues but may be expressed under abnormal conditions. Moreover, insulin receptor substrate-5 (DOK4) and insulin receptor substrate-6 (DOK5) are linked to lymphocyte regulation. An improved understanding of the signaling pathways mediated by IRS proteins in hematopoiesis-related processes, along with the increased development of agonists and antagonists of these signaling axes, may generate new therapeutic approaches for hematological diseases. The scope of this review is to recapitulate and review the evidence for the functions of IRS proteins in normal and malignant hematopoiesis.

The correlation between level of serum APPL1 and urinary albumin excretion rate in patients with type 2 diabetes mellitus / 中华内分泌代谢杂志

Objective To investigate the association of adaptor protein,phosphotyrosine interacting with PH domain and leucine zipper 1(APPL1)with urinary albumin excretion rate in patients with type 2 diabetes mellitus (T2DM),and to explore the role of APPL1 in the development of diabetic kidney disease(DKD). Methods According to the urinary albumin/creatinine ratio(UACR),288 newly-diagnozed patients with T2DM were divided into normal albuminuria group(UACR<30 mg/g,n=116),microalbuminuria group(UACR 30 ~300 mg/g,n=95),and macroalbuminuria group(UACR>300 mg/g,n=77). 130 healthy subjects with matched sex and age were used as control group. Serum APPL1,tumor necrosis factor α(TNF-α),and adiponectin levels were measured by ELISA method. Results Serum APPL1 level in T2DM patients was significantly higher than that in control subjects (P<0.01), and increased with the rising of UACR. In patients with T2DM, serum APPL1 level was negatively correlated with estimated glomerular filtration rate(r=-0.246, P<0.01) while it was positively correlated with HbA1C, low density lipoprotein cholesterol, total cholesterol, triglycerides, insulin resistance index, serum creatinine,blood urea nitrogen, systolic blood pressure, TNF-α, and adiponectin(r=0. 119, 0. 167, 0. 209, 0.194,0.273,0.242,0.131,0.144,0.365, and 0.952, respectively, P<0.05 or P<0.01). Conclusion Serum APPL1 level in patients with T2DM was increased with the rising of UACR, suggesting that APPL1 may be involved in the development of DKD.

Relationship between inflammatory responses and autophagy in lung tissues after scald and NOD2 signaling pathway in septic rats / 中华麻醉学杂志

Objective To evaluate the relationship between inflammatory responses and autophagy in lung tissues after scald and nucleotide-binding oligomerization domain-containing protein 2 (NOD2) signaling pathway in septic rats.Methods Twenty SPF healthy male Sprague-Dawley rats,weighing 200-250 g,were divided into control group (group C,n=10) and sepsis after scald group (group SS,n=10) using a random number table.The rats were subjected to a third-degree scald burn covering 20％ of total body surface area (body surface was shaved and then exposed to 99-100 ℃ water for 12 s),and 24 h later muramyldipeptide 5 mg/kg was intravenously injected to induce sepsis.The rats were only exposed to 20 ℃ water,and 24 h later normal saline 1 ml was given instead in group C.At 6 h after muramyldipeptide injection in group SS and at 6 h after normal saline injection in group C,arterial blood samples were collected for determination of serum tumor necrosis factor-r and interleukin-6 concentrations by enzyme-linked immunosorbent assay.Then rats were sacrificed and lungs were removed tor measurement of activity of myeloperoxidase,NOD2 mRNA expression (using real-time polymerase chain reaction) and expression of receptor interacting protein 2,nuclear factor kappa Bp65 and microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ) and LC3 Ⅱ in lung tissues (by Western blot).The LC3 Ⅱ / Ⅰ ratio was calculated.Results Compared with group C,the expression of NOD2 mRNA,receptor interacting protein 2 and nuclear factor kappa Bp65 was significantly up-regulated,and the LC3 Ⅱ / Ⅰ ratio and serum tumor necrosis factor-α and interleukin-6 concentrations were increased in group SS (P＜0.05).Conclusion The mechanism underlying enhanced inflammatory responses and autophagy in lung tissues during sepsis after scald may be related to activation of NOD2 signaling pathway in rats.

Expression of Grb2, MMP-3, and MMP-9 in cholangiocarcinoma and its significance / 临床肝胆病杂志

ObjectiveTo investigate the expression of growth factor receptor-bound protein-2 (Grb2), matrix metalloproteinase-3 (MMP-3), and MMP-9 in cholangiocarcinoma and its significance. MethodsThe expression of Grb2, MMP-3, and MMP-9 in cholangiocarcinoma tissues of 47 cases and normal tissues was measured using immunohistochemistry, and the correlations of Grb2 expression with clinical pathology and MMP-3 and MMP-9 expression were analyzed. Comparison of continuous data was made using t test, and the correlation of Grb2 expression with MMP-3 and MMP-9 expression was analyzed using the multivariate linear regression model. ResultsThe expression of Grb2 in cholangiocarcinoma tissues was significantly higher than that in normal bile duct tissues (t=5935, P＜0.001); the expression of Grb2 in cholangiocarcinoma tissues and normal bile duct tissues showed no significant correlation with age, sex, and differentiation level; the expression of Grb2 in cholangiocarcinoma tissues with lymph node or distant metastasis was significantly higher than that in cholangiocarcinoma tissues without metastasis (t=3.882, P=0.003). The expression of Grb2 was positively correlated with the expression of MMP-3 and MMP-9 (r2=0.3667, P=0.018; r2=0.5133, P=0.007). ConclusionThe expression of Grb2 in cholangiocarcinoma tissues is higher than that in normal bile duct tissues, and it is closely related to the invasion and metastasis of carcinoma. Further study shows that the expression of Grb2 is positively correlated with the expression of MMP-3 and MMP-9.

Research of XB130,a novel adaptor protein in cancer / 实用肿瘤学杂志

Currently,XB130 as a newly discovered characterized adaptor protein ,it has been implicated as a substrate and regulator of many intracellular signal transduction ,such as FAK/SRC,PI3K/Akt and MEK-ERK signaling and so on.It has been found that XB130 is high expression in many cell lines ,for instance thyroid carcinoma,osteosarcoma,gastric cancer,esophageal cancer and breast cancer etc .The mechanism of XB130 in tumor is becoming increasingly attention .XB130 is recently attributed to be a new oncogene ,and plays important roles in cell pro -liferation,cell survival and tumorigenesis .A deeper understanding of these mechanisms may lead to the discovery of XB130 as an important mediator in tumor development and as a novel therapeutic target for cancer.

Location of recombinant adenovirus Ad5/F35-HF2S and its interaction with Bcr-Abl in chronic myeloid leukemia K562 cells / 肿瘤

Objective: To testify the location of recombinant adenovirus Ad5/F35-HF2S in chronic myeloid leukemia K562 cells and its interaction with the fusion protein Bcr-Abl. Methods: The recombinant adenovirus Ad5/F35-HF2S was infected into K562 cells, then the expression of fusion protein HF2S in K562 cells was assessed by RT-PCR (reverse transcription-PCR) and Western blotting. The location of HF2S protein in K562 cells was demonstrated by immunofluorescence and Western blotting assay. The immunofluorescence was performed to verify the co-localization of HF2S and Bcr-Abl. The co-immunoprecipitation assay was employed to detect the interaction between HF2S and Bcr-Abl. Results: The leukemia cell line K562 was effectively infected by the adenovirus Ad5/F35-HF2S. The fusion protein HF2S and Bcr-Abl were co-located and interacted with each other in the cytoplasm. Conclusion: HF2S can be successfully expressed and interacted with Bcr-Abl in the cytoplasm of K562 cells. These results provide the basement of the targeted therapy for chronic myeloid leukemia. Copyright © 2013 by TUMOR.

Regulatory role of acetylcholine receptor in muramyl dipeptide-induced activation of NLR2/RIP2 pathway in macrophages of mice / 中华麻醉学杂志

Objective To evaluate the regulatory role of acetylcholine receptor in muramyl dipeptide (MDP)-induced activation of Nod-like receptor 2/receptor-interacting protein 2 (2NLR2/RIP2) pathway in macrophages of mice.Methods RAW264.7 cells at the logarithmic growth phase were seeded in 12-well plates (density 1 × 106 cells/ml,2 ml/well),a total of 108 wells.The cells were randomly divided into 3 groups (n =36 each) using a random number table:control group (group C),MDP group (group M),and GTS-21 (a7nAChR specific agonist) group (group G).The cells were routinely cultured in group C.MDP with the final concentration of 10 μg/ml was added to the culture medium in group M.MDP with the final concentration of 10μg/ml and GTS21 with the final concentration of 50 μg/ml were added to the culture medium in group G.The cells were incubated for 24 h.At 1,6 and 24 h of incubation with MDP,12 wells were chosen and the cell suspension was obtained for measurement of NLR2 mRNA expression (by real-time fluorescent quantitative PCR),RIP2 expression (by Western blot),and concentrations of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) in the culture media (by ELISA).Results Compared with group C,the levels of NLR2 mRNA,RIP2,TNFα and HMGB1 were significantly increased at each time point in group M (P ＜ 0.05).Compared with group M,the levels of NLR2 mRNA,RIP2,TNF-α and HMGB1 were significantly decreased at each time point in group G (P ＜ 0.05).Conclusion Acetylcholine receptor can suppress MDP-induced transduction of NLR2/RIP2 pathway in macrophages of mice.

Role of growth factor receptor-bound 2 in homo sapiens eukaryotic translation elongation factor 1 alpha 2 promoting the progenesis and development of pancreatic cancer / 中华消化杂志

Objective To explore the role of growth factor receptor-bound 2 (GRB2) in homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) promoting the carcinogenesis and development of pancreatic cancer.Methods The human pancreatic cancer cell line SW1990 with low expression of EEF1A2,was transfected by Ad5/F35-EEF1A2 plasmid.The expression of EEF1A2 in human pancreatic cancer cell lines BxPC-3 cells with high expression of EEF1A2,was down-regulated by small interfering RNA (siRNA) interference.The expressions of GRB2 in SW1990 and BxPC-3 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after interference.The pancreatic cancer BxPC-3 cells were interfered by specific and efficient chemical synthesized siRNA fragment.The changes of BxPC-3 cell proliferation and migration were observed by methyl thiazoly tetrazolium (MTT) and Transwell assay before and after interference.The data were analyzed by one-way analysis of variance.Results After human pancreatic cancer cell line SW1990 was transfected with Ad5/F35-EEF1A2 plasmid,the expression of GRB2 at mRNA and protein level both increased significantly (F=5.67,6.39,both P＜0.05).After the expression of EEF1A2 was inhibited by EEF1A2-siRNA,the expression of GRB2 at mRNA and protein level both decreased significantly (F=6.59,4.69,both P＜0.05).After BxPC-3 cells was transfected with siRNA-GRB2 for 72 hours,the results of MTT assay indicated that the absorbance value was 1.22±0.14,compared with negative control group (1.42±0.15) and blank control group (1.39 ± 0.15) the difference was statistically significant (F =6.63,P＜ 0.05).The results of transwell assay showed that the migration ability of cells in siRNA-GRB2 group decreased.After 24 hours,the number of migrated cells was 24.10±4.25,compared with negative control group (54.72±5.24) and blank control group (51.26 ± 6.23) the difference was statistically significant (F=55.00,P＜ 0.05).Conclusion GRB2 is key molecule in EEF1A2 promoting the progenesis and development of pancreatic cancer.

Effect of remifentanil on nucleotide-binding oligomerization domain 1 mRNA expression in rats with renal ischemia-reperfusion injury / 中华麻醉学杂志

Objective To investigate the effect of remifentanil on nucleotide-binding oligomerization domain 1 (NOD1) mRNA expression in rats with renal ischemia-reperfusion (I/R) injury.Methods Sixty male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each):sham operation group (S group),I/R group and remifentanil group (R group).Renal ischemia was induced by occlusion of bilateral renal arteries for 45 min followed by 24 h reperfusion in groups I/R and R.Remifentanil 1.0 μg· kg-1 · min-1 was infused until 30 min of reperfusion starting from 15 min before ischemia in group R,while the equal volume of normal saline was given instead in S and I/R groups.The animals were sacrificed at 15 min before ischemia and at 3,6,24 h of reperfusion and the kidneys were removed for microscopic examination and polymorphonuclear leukocyte (PMN) count and for measurement of NOD1 mRNA expression (by RT-PCR).The apoptotic rate was determined by flow cytometry double staining method.Results Compared with group S,NOD1 mRNA expression was up-regulated,and the apoptotic rate and PMN count were significantly increased at each time point during reperfusion in group I/R,and the apoptotic rate and PMN count were significantly increased at each time point during reperfusion,and NOD1 mRNA expression was up-regulated at 6 and 24 h of reperfusion in group R (P ＜ 0.01).Compared with I/R group,NOD1 mRNA expression was down-regulated,and the apoptotic rate and PMN count were significantly decreased at each time point during reperfusion (P ＜ 0.05 or 0.01),and the pathological changes were significantly attenuated in group R.Conclusion Remifentanil can reduce the renal I/R injury by down-regulating the expression of NOD1 mRNA and inhibiting inflammatory response and apoptosis.

Influence of Grb2-SH3 inhibitor peptidimer-c on the apoptosis-related genes expression in K562 cells / 肿瘤研究与临床

Objective To analyze the influence of peptidimer-c on the gene expression profiling of K562 cells and investigate the mechanism of peptidimer-c inducing the apoptosis and inhibiting proliferation of K562cells.Methods Trypan blue staining technique was performed for counting the number of living K562 cells treated with peptidimer-c.The ultrastructure changes of K562 cells treated with peptidimer-c was observed under transmission electron microscope.The Human U133 Plus 3.0 gene chips were used to detect the differentially expressed genes of K562 cells treated with peptidimer-c.Reverse transcription PCR was conducted to confirm some genes identified by gene chips.Results Peptidimer-c could induce the apoptosis and inhibit the proliferation of K562 cells.Peptidimer-c caused widely changes of the gene expression profiles of K562 cells.The chip data suggested that there were 529 differentially expressed genes,of which 455 genes were up-regulated and 74 genes were down-regulated.The relevant apoptotic genes were down-regulated markedly,including JUN,AXUD1,TNFRSF10B,etc.Fifteen of the differentially expressed genes were detected by RT-PCR,which was consistent with the chip data.Conclusion Peptidimer-c may induce aooptosis of K562 cells by activating the TNF/TNFR family and the JUN family.

Research on the association of TIRAP coding region polymorphism with susceptibility to tuberculosis in Chinese Han population / 中华微生物学和免疫学杂志

Objective To detect specific polymorphisms in Toll-interleukin 1 receptor domain containing adaptor protein(TIRAP) coding region for Chinese Han population, and verify whether they are associated with susceptibility to tuberculosis. Methods Search TIRAP polymorphisms by sequencing in small sample; detect single nucleotide polymorphism(SNP) by ligase detection reaction technique in large sample; analyze whether polymorphisms are related to tuberculosis by statistic methods. Results Four polymorphisms were present in the TIRAP coding region. 394A had higher frequencies in the tuberculosis(TB)group than the control. But allelic and genotypic analysis showed that there were no significant difference in statistic between TB patients and controls(P>0.05). The SNP G164A mutation related with TB patient's condition. Comparing to controls, retreatment patients' allelic frequencies had significant difference in statistic(P<0.05), sputum positive patients and lung cavitation patients had lower 164A frequencies. Conclusion TIRAP coding region polymorphisms may be risk factors for TB occurrence and development in Chinese Han population.

Transmembrane Adaptor Proteins Positively Regulating the Activation of Lymphocytes

Engagement of the immunoreceptors initiates signaling cascades resulting in lymphocyte activation and differentiation to effector cells, which are essential for the elimination of pathogens from the body. For the transduction of these immunoreceptor-mediated signals, several linker proteins termed transmembrane adaptor proteins (TRAPs) were shown to be required. TRAPs serve as platforms for the assembly and membrane targeting of the specific signaling proteins. Among seven TRAPs identified so far, LAT and LIME were shown to act as a positive regulator in TCR-mediated signaling pathways. In this review, we will discuss the functions of LAT and LIME in modulating T cell development, activation and differentiation.

Enfermedad inflamatoria intestinal: Una mirada inmunológica: [revisión] / Inflammatory bowel diseases: An immunological approach: [review]

Inflammatory bowel diseases (IBD) are inflammatory diseases with a multifactorial component that involve the intestinal tract. The two relevant IBD syndromes are Crohn's disease (CD) and ulcerative colitis (UC). One factor involved in IBD development is a genetic predisposition, associated to NOD2/CARD15 and Toll-like receptor 4 (TLR4) polymorphisms that might favor infectious enterocolitis that is possibly associated to the development of IBD. The identification of specific immunologic alterations in IBD and their relationship to the etiology of the disease is a relevant research topic. The role of intra and extracellular molecules, such as transcription factors and cytokines that are involved in the inflammatory response, needs to be understood. The relevance of immunologic molecules that might drive the immune response to a T helper (Th) 1, Th 2 or the recently described Th 17 phenotype, has been demonstrated in animal models and clinical studies with IBD patients. CD and UC predominantly behave with a Th 1 and Th 2 immune phenotype, respectively. Recently, an association between CD and Th 17 has been reported. The knowledge acquired from immunologic and molecular research will help to develop accurate diagnostic methods and efficient therapies.

Mechanism of epileptic seizures in the neuron-specific adaptor protein complex type 3B knockout mice / 中华神经科杂志

Objective To explore the mechanism of spontaneous seizures in adaptor protein complex type 3B knockout mice(AP3M2KO mice).Methods AP3M2KO mice were generated.Seizures and electroencephalogram(EEG)were monitored using video camera and telemetry system.Glutamate and GABA releases were determined using in vivo microdialysis method.Results AP3M2KO mice began to suffer from spontaneous seizures 8 weeks after the birth,but did not show any other behavior abnormality.The onset of ictal discharge over the temporal region was synchronized with seizures.There were no significant differences in basal glutamate and GABA releases in hippocampus between AP3M2KO((0.35±0.08)pmol/20μl and(2.94±1.69)fmol/20μl,respectively)and wild-type mice.However,the 50 mmol/L K+-evoked GABA release was impaired in AP3M2KO mice((63.5±11.8)fmol/20μl vs(209.2±63.7)fmol/20 μl,t=4.405,P＜0.05),whereas no significant difference was found in K+-evoked glutamate release.Conclusions AP3M2KO mice suffer from epileptic seizures similar to the clinical features of human epilepsy.The impairment of inhibitory GABAergic transmission iS involved in the mechanism of spontaneous seizures in AP3M2KO mice.

Changes of expression of FADD and Daxx following focal cerebral ischemia in rats / 中华神经科杂志

Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

Progress of Lnk adaptor protein / 医学研究生学报

Lnk is a member of a newly discovered adaptor protein family.It negatively regulates the Jak-STAT,Ras/MAPK,PI3K signaling pathways.Lnk plays a critical role in hematopoiesis,endothelial cell activation and actin cytoskeleton regulation.This article aims to review the progress of Lnk adaptor protein.