Objective To investigate the variation and correlation between tumor necrosis factor-?(TNF-?)and transforming growth factor-?1(TGF-?1)in children with Henoch-Schonlein purpura nephritis(HSPN)in plasma and explore their effects on kidney lesion in children with Henoch-Schonlein purpura(HSP).Methods Plasma TNF-? and TGF-?1 were determined with enzyme-linked immunosorbent assay(ELISA)in 30 cases with HSP,38 cases with HSPN and 30 normal controls,urinary protein excretion with urinary analyze method in these children.Renal biopsies were performed and renal biopsy specimens were observed by light,immunofluorescence and electron microscopy in 32 out of 38 cases with HSPN.The SPSS 11.0 software was used to analyze the data.Results 1.Comparing with normal controls,the plasma level of TNF-? and TGF-?1 in children with HSP increased with significant difference in statistics(Pa

Objective To examine the expression and analyze the significance of autophagy-related gene microtubule-associated protein 1 light chain 3 (MAP1-LC3,LC3),p62 and lysosorne-associated membrane protein 2 (LAMP-2) in pancreatic tissues of mice with acute pancreatitis (AP).Methods Twenty mice were randomized into AP group and control group,and the number of mice was equal between two groups.AP group was intra-peritoneally injected by 20％ L-arginine solution (two injections of 4 g/kg body weight,every 1 h) in the dosage of 4 g/l kg twice every 1 hour to establish AP model,while control group was administered with equal volume of normal saline by intra-peritoneal injection.All the mice were euthanized at 24 hour after the last injection.Pancreatic histopathological changes were measured.In addition,the protein expressions of LC3,p62 and LAMP-2 were detected by Western blot.Results No obvious pathological changes were observed in control group.Pancreatic acinar edema,structure destruction,missing,the obvious widening of interlobular septum,small interlobular septum and acinar septum,and the necrosis of acinar cells at different degrees were observed in AP group.The pathological score for tissue edema,hemorrhage,necrosis and inflammation in AP group was 3.13 ± 0.50,2.83 ± 0.32,3.25 ± 0.46 and 3.16 ± 0.47,respectively,which was all 0 in control group.The differences were statistically significant between AP group and control group (P ＜ 0.01).In AP group,the ratio of LC3-Ⅱ/LC3-Ⅰ,p62 and LAMP-2 protein in pancreatic tissue were 1.16 ± 0.08,0.94 ± 0.04 and 0.35 ± 0.04,respectively,which were 0.24 ± 0.02,0.34 ± 0.03 and 0.95 ± 0.03 in control group.The ratio of LC3-Ⅱ/LC3-Ⅰ and p62 protein in pancreatic tissue in AP group were much higher than those in control group,while LAMP-2 in AP group was lower than that in control group,and there was statistically significant difference between two groups (all P ＜0.01).Conclusions Intraperitoneal injection of L-arginine could induce acute pancreatitis,and autophagy is impaired,which was associated with decreased LAMP-2 protein expression.

Objective To explore the alteration and effect of autophagy in pancreas tissue of rat injected by exenatide.Methods Diabetes model rats were induced by two-month high-sugar and high-fat diet and streptozotocin injection (35 mg/kg) in normal rats.50 SD male rats were divided into four groups according to the principle of complete random design,namely normal control group (n=10),normal exenatide-injected group (n=10),diabetes-model control group (n=l5) and diabetes-model exenatide-injected group (n=15).Rats in exenatide-injected groups were subcutaneously injected with exenatide respectively in 5 μg/kg dose each time,twice a day,at 8 a.m.and 6 p.m.Animal weights were weighted weekly and the dose of exenatide was adjusted according to current weight.Rats in the two control groups were injected with the corresponding amount of saline.Mter 10 weeks of treatment,all rats were killed and pancreatic tissues were disposed.Immunohistochemistry was used to measure the expression of GLP-1R in pancreatic tissues.Western blot was used to test the expressions of LC3-Ⅰ,LC3-Ⅱ and p62 in pancreatic tissues,and LC3-Ⅱ/Ⅰ ratio and p62 were compared between any two groups.All specimens were stained with hematoxylin-eosin (HE).The data were expressed as means ± standard deviation and were analyzed by unpaired Student t test using SPSS 18.0 statistics software.P value ＜0.05 was considered to be statistically significant for all tests.Results The pancreatic tissues from 13 rats (6 from the normal exenatide-injected group and 7 from the diabetes-model exenatide-injected group) appeared pathological changes such as gland structure damage,pancreatic cells atrophy and cells compartment broadening.The expressions of GLP-1R,LC3-Ⅱ and p62,and LC3B-Ⅱ/Ⅰ ratio in the two exenatide-injected groups were higher than those in the respective control group,and the differences had statistical significance (P＜0.05).Conclusions Long-term subcutaneous injection of exenatide can upregulate the expression of GLP-1R in rat pancreatic acinar cells and may induce the impaiment of autophagy flux in rat pancreatic cell.

BACKGROUNDTransient sublethal ischemia is known as ischemic preconditioning, which enables cells and tissues to survive subsequent prolonged lethal ischemic injury. Ischemic preconditioning exerts neuroprotection through phosphatidylinositol 3-kinase (PI3K)/Akt pathway. Cbl-b belongs to the Casitas B-lineage lymphoma (Cbl) family, and it can regulate the cell signal transduction.The roles of ubiquitin ligase Cbl-b and PI3K/Akt pathway and the relationship between them in oxygen-glucose deprivation preconditioning (OGDPC) in PC12 cells were investigated in the present study.

METHODSOxygen and glucose deprivation (OGD) model in PC12 cells was used in the present study. The 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, nuclear staining with Hoechst 33258, and Western blotting were applied to explore the roles of Cbl-b and PI3K/Akt pathway and the relationship between them in OGDPC in PC12 cells.

RESULTSCell viability was significantly changed by OGD and OGDPC. OGD significantly decreased cell viability compared with the control group (P < 0.05), and preconditioning could rescue this damage was demonstrated by the increase of cell viability (P < 0.05). The expression of Cbl-b was significantly increased after OGD treatment. However, the activation of Akt and GSK3β was greatly inhibited. Preconditioning could inhibit the increase of Cbl-b caused by OGD and increase the activation of Akt and GSK3β. LY294002, a specific inhibitor of PI3K, could effectively inhibit the increase of Akt and GSK3β after preconditioning treatment. It partly inhibited the decrease of Cbl-b expression after preconditioning treatment.

CONCLUSIONUbiquitin ligase Cbl-b and PI3K/Akt pathway are differently involved in OGDPC in PC12 cells.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Survival ; Glucose ; deficiency ; Ischemic Preconditioning ; Oxygen ; metabolism ; PC12 Cells ; Phosphatidylinositol 3-Kinase ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Proto-Oncogene Proteins c-cbl ; genetics ; metabolism ; Rats ; Signal Transduction ; physiology

OBJECTIVETo analyze the changes of 2009 FIGO staging system compared with the 1988 FIGO staging system of endometrial carcinoma and evaluate the diagnostic value of MRI staging by the 2009 FIGO criteria.

METHODSA retrospective study was performed on 63 consecutive patients with pathologically confirmed endometrial carcinoma who were treated by surgery initially from January to December 2009. The diagnostic value of preoperative MRI by the 2009 FIGO staging system was compared with that using the 1988 FIGO system, respectively.

RESULTSAccording to the 2009 FIGO staging system of endometrial carcinoma, stage Ia was defined as no or less than half myometrial invasion, which included stage Ia (confined to endometrium) and stage Ib (invasion less than half of the myometrium) of the 1988 FIGO staging system. Stage Ib assessed by the 2009 FIGO system was the same as the stage Ic of 1988 FIGO system, indicating the lesions more than half myometrial invasion. Endocervical glandular involvement only (stage IIa of 1988 FIGO system) was classified as stage I. Positive cytology of ascites (stage IIIa of 1988 FIGO system) was excluded by the 2009 FIGO criteria. Using the 1988 FIGO system, the accuracy of MRI for the evaluation of endometrial carcinoma of stage Ia, Ib, Ic, whole stage I, IIa, IIb, whole stage II, IIIa, IIIb, IIIc, whole stage III and IVb were 95.2%, 79.4%, 81.0%, 84.1%, 96.8%, 90.5%, 90.5%, 92.1%, 98.4%, 92.1%, 82.5%, and 98.4%, respectively, while using the 2009 FIGO system, the accuracy of MRI of stage Ia, Ib, whole stage I, II, IIIa, IIIb, IIIc, whole stage III and IVb were 88.9%, 81.0%, 88.9%, 92.1%, 98.4%, 98.4%, 92.1%, 88.9% and 98.4%, respectively.

CONCLUSIONSThe 2009 FIGO staging system is simplified on the basis of the 1988 FIGO system. It gives an improved accuracy of MRI in evaluating the stage I to III endometrial carcinoma.

Adenocarcinoma ; diagnosis ; pathology ; surgery ; Adult ; Aged ; Endometrial Neoplasms ; diagnosis ; pathology ; surgery ; Female ; Humans ; Hysterectomy ; International Agencies ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Myometrium ; pathology ; Neoplasm Invasiveness ; Neoplasm Staging ; methods ; Preoperative Period ; Retrospective Studies

OBJECTIVETo investigate the association of HLA class I and II alleles with generalized vitiligo in ethnic Han Chinese in north China.

METHODSBy employing polymerase chain reaction sequence-specific primer (PCR-SSP) procedure 34 generalized vitiligo patients in north China were studied for HLA I and II alleles and were compared with 102 healthy controls.

RESULTSThe allelic frequencies of HLA-A*30, Cw*06, DRB1*07, and DQB1*0201 were increased significantly in generalized vitiligo and especially in the patients without family history compared with the controls.

CONCLUSIONThese alleles positively associated with generalized vitiligo in Chinese Han patients in north China, might provide clues to reveal the susceptibility gene(s) of vitiligo in Chinese and as well as the immunnogenetic mechanisms of disease.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genes, MHC Class I ; genetics ; Genes, MHC Class II ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Vitiligo ; ethnology ; genetics

OBJECTIVETo investigate the possible mechanism of local hyperthermia in the treatment of warts through detecting the differences in CD1a/CD83 of Langerhans cells (LCs) in émigrés from HPV-infected skin, as compared to normal skin.

METHODSConfocal microscopy were performed on Condyloma Accuminatum (CA)and normal skin; Freshly taken biopsies of CA and normal skin were subjected to surface heating at 37 degrees C, 42 degrees C and 45 degrees C respectively, for 30 mins. Flow cytometry was used to determine the CD1a/ CD83 changes of LCs in émigrés from CA and normal skin.

RESULTSBy confocal microscopic observation, there were practically no CD1a+ LCs that expressed CD83 in the epidermis of both normal skin and CA. The proportions of CD1a+/CD83 LCs were significantly increased with increased temperatures in émigrés from both normal skin and CA. At each given temperature, the numbers of LCs in émigrés from CA were greater than those from normal skin.

CONCLUSIONLocal hyperthermia can promote migration and maturation of LCs in HPV-infected skin and accordingly stimulate the immune system to treat warts.

Adult ; Cell Movement ; immunology ; Female ; Humans ; Hyperthermia, Induced ; In Vitro Techniques ; Langerhans Cells ; cytology ; immunology ; Papillomaviridae ; immunology ; pathogenicity ; Skin ; immunology ; virology ; Young Adult

BACKGROUNDThe sensitization and elicitation phases are involved in the immunopathogenesis of contact hypersensitivity (CHS). Langerhans cells (LCs) are believed to play pivotal roles in the sensitization stage of CHS. Local hyperthermia on skin induces the migration as well as maturation of epidermal LCs. Although fever-range whole body hyperthermia and local hyperthermia at 43°C prior to sensitization were reported to suppress CHS, the effects of different temperatures and the timing sequence of local hyperthermia on CHS have not been tackled.

METHODSLocal hyperthermia was applied to murine dorsal skin 3 days prior to, concurrent with, or 2 days post sensitization with fluorescein isothiocyanate (FITC) in BALB/c mice. Local hyperthermia temperatures at 37°C, 39°C, 41°C and 43°C were applied to mouse dorsal skin and the severity of CHS was calculated by measuring the swelling response of the challenged ears.

RESULTSLocal hyperthermia at 39°C, 41°C and 43°C prior to sensitization reduced the severity of CHS, as compared with that at 37°C. The suppression of CHS was temperature dependent in that higher temperature had a stronger effect. On the contrary, the hyperthermia treatments, either concurrent with or post-sensitization, resulted in an enhanced temperature-dependent ear swelling response.

CONCLUSIONSThe severity of murine CHS could be influenced by local hyperthermia at the sensitization stage in a temperature dependent manner. The temporal effect of local hyperthermia suggested a novel factor in interpreting the severity of allergic contact dermatitis.

Animals ; Dermatitis, Contact ; therapy ; Female ; Hyperthermia, Induced ; Langerhans Cells ; physiology ; Mice ; Mice, Inbred BALB C

BACKGROUNDAtopic dermatitis (AD) is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 (Th1/Th2) cytokine expression. Filaggrin (FLG) is the key protein to maintaining skin barrier function. Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes. However, the role of Th1/Th2 cytokines on FLG processing is not substantially documented. Our aim was to investigate the impact of Th1/Th2 cytokines on FLG processing.

METHODSHaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin (IL)-4, 13 or interferon-γ (IFN-γ). FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor (LEKTI) were measured by both real-time quantitative polymerase chain reaction and Western blotting. Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.

RESULTSIL-4/13 significantly reduced, while IFN-γ significantly up-regulated FLG expression. IL-4/13 significantly increased, whereas IFN-γ significantly decreased the expression of kallikreins 5 and 7, matriptase and channel-activating serine protease 1. On the contrary, IL-4/13 significantly decreased, while IFN-γ increased the expression of LEKTI and caspase-14. Similar trends were observed in AD lesions.

CONCLUSIONSOur results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes. The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.

Caspase 14 ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Dermatitis, Atopic ; metabolism ; Humans ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Intermediate Filament Proteins ; metabolism ; Keratinocytes ; enzymology ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; metabolism ; Serine Peptidase Inhibitor Kazal-Type 5 ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

OBJECTIVETo evaluate the ability of a polycaprolactone/polylactic acid (PCL/PLA) membrane to inhibit epidural scar adhesion after laminectomy, and observe the responsive changes of the pain media in the spinal cord.

METHODSL(1), L(3) laminectomies were performed on 96 Wistar rats. The rats were divided into 3 groups: None-implant Control Group (NC), Autologous free fat graft group (AFFG) and PCL/PLA membrane group (PCL/PLAm). The rats were killed at 1, 3, 6, and 12 weeks postoperatively. Epidural scar formation and adhesion were observed grossly and histologically. Reverse transcription polymerase chain reaction (RT-PCR) were used to analyses the expression of Transforming growth factor beta (TGF-beta) in the epidural scar. Immunohistochemistry stain and RT-PCR were performed to evaluate the expression of the substance P and the c-fos gene in the relevant spinal cord, and the results were analyzed statistically.

RESULTSGross evaluation and histological evaluation showed that in the NC lamina defect site had much scar tissue and had wide and tight adhesions to the dura; in the AFFG, with the fat degrading gradually, the adhesions were increased; whereas in the PCL/PLAm group, there were slightly adhesions to the dura. RT-PCR showed that the expression of the TGF-beta was much less in the PCL/PLAm group than in the NC group. The insertion of the PCL/PLA membrane and the fat patch reduced the expression of the substance P and the c-fos gene in the spinal cord.

CONCLUSIONThe insertion of the PCL/PLA membrane reduces scar formation and separates fibrosis tissue from the dura, the results indicate that PCL/PLA membrane is an effective way of reducing peridural scar formation and preventing the failed back surgery syndrome.

Animals ; Biocompatible Materials ; Cicatrix ; prevention & control ; Female ; Lactic Acid ; Laminectomy ; adverse effects ; Membranes, Artificial ; Polyesters ; Polymers ; Postoperative Complications ; prevention & control ; Prosthesis Implantation ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Rats ; Rats, Wistar ; Spinal Cord ; metabolism ; Spinal Diseases ; prevention & control ; Substance P ; biosynthesis ; Tissue Adhesions ; prevention & control