Objective:Prepared the specific chicken egg yolk immunoglobulins (IgY) against the cell wall protein of Trichophyton mentagrophytes (tmCWP) and detected its biological activities,which was to establish the basis for the preventment and treatment in dermatophytes disease.Methods: In this work,tmCWP was extracted and purified by cold alkali method,and being used as immunogen to immunized healthy laying hens.The IgY was extracted from the egg yolk by polyethylene glycol method and purified by saturated ammonium sulfate method,respectively.The concentration of the extracted IgY was detected by Bradford method.The purity and molecular weight of the specific anti-tmCWP IgY were analysed by SDS-PAGE.The titer of IgY was obtained by ELISA.The immunoreactivity of IgY was performed by Western blot.Results: The purity of the extracted IgY reached to 87.27％.ELISA indicated that the titer of the specific anti-tmCWP IgY gradual rised 20 days after primary immunization and reached to the highest value (1∶32 000) after 45 days.Western blot revealed that the specific IgY showed a good immunoreactivity and a specifically combination capacity.Conclusion: In our work,the tmCWP could be regarded as the immunogen to prepare the specific anti-tmCWP IgY,which could provide a novel thought for the therapy of Trichophyton mentagrophytes infection.

From August 2011 to March 2012,5 241 type 2 diabetic patients with body mass index ≥ 24kg/m2 were enrolled from 60 hospitals in Guangdong Province.According to estimated glomerular filtration rate (eGFR),a total of 2 631 subjects with norml urine microalbumin level (＜30 ng/L) were divided into normal eGRF group and decreased eGRF group.Binary logistic regression was used to analyze the associations between eGFR and its related risk factors.The results showed that age,blood uric acid,blood urea nitrogen,history of hypertension and coronary heart disease,family history of diabetes,and hyperuricemia were positively related to lowering of eGFR (P＜ 0.05 or P＜0.01).HbA1C＜7％,regular glucose monitoring,and regular physical activity were negatively related to eGFR decrease (all P＜ 0.01).These results suggest that urine microalbumin and eGFR should be applied to overweight or obese patients with type 2 diabetes in order to screen diabetic nephropathy.Furthermore,intensive control of blood glucose,uric acid,and blood pressure is beneficial to lowering the risk of diabetic nephropathy.

Objective To evaluate effects of a fusion protein LTβR-Fc, which can block the herpesvirus entry mediator ligand (LIGHT-HVEM)signaling pathway, on ovalbumin-induced dermatitis in a mouse model. Methods Thirty BALB/c mice were randomly and equally divided into 3 groups: blank control group treated with 100 μl of sodium chloride physiological solution, model group sensitized with 100 μl of sodium chloride physiological solution containing 100 μg ovalbumin, blocker group firstly blocked with 100 μl of sodium chloride physiological solution containing 100 μg LTβR-Fc followed by sensitization with 100 μl of sodium chloride physiological solution containing 100 μg ovalbumin at 24 hours after the blocking. Disease severity was evaluated by eczema area and severity index (EASI)score, and lesional size was measured on day 0, 4, 8, 12, 15, 20, 23, 27, 31 and 34 after the first sensitization. A total of three sessions of sensitization were carried out. At the end of treatment, all the mice were sacrificed after serum was obtained from their orbital cavities. Thereafter, tissue specimens were obtained from skin lesions, and single cell suspensions of the spleen were prepared. RT-PCR was performed to detect mRNA expressions of interferon γ (IFN-γ), interleukin 4 (IL-4)and IL-5 in murine lesions, ELISA to measure IFN-γ, IL-4 and IL-5 levels in culture supernatants of murine splenocytes, as well as ovalbumin-specific and total IgE and IgG1 levels in murine sera. Results LTβR-Fc significantly suppressed inflammatory response in the mouse model of dermatitis induced by ovalbumin. Compared with the model group, the blocker group showed significantly decreased lesion area and EASI score (both P < 0.05). In addition, a significant decrease was observed in the mRNA expressions of IL-4 (0.88 ± 0.25 vs. 1.81 ± 0.25, P < 0.05), IL-5 (0.75 ± 0.15 vs. 1.24 ± 0.26, P < 0.05)and IFN-γ (0.62 ± 0.09 vs. 1.11 ± 0.19, P < 0.05)in murine lesions, and in supernatant levels of IL-4 (9.58 ± 1.44 ng/L vs. 20.12 ± 5.39 ng/L, P < 0.05), IL-5 (11.37 ± 2.02 ng/L vs. 22.77 ± 4.07 ng/L, P < 0.05)and IFN-γ (16 167 ± 950.40 ng/L vs. 23 930 ± 44.20 ng/L, P < 0.05)in the blocker group compared with the model group. The serum levels of both total IgE and ovalbumin-specific IgE were significantly lower in the blocker group than in the model group(total IgE: 27 466.67 ± 2 052.64 μg/L vs. 32 277 ± 407.53 μg/L, P < 0.05; ovalbumin-specific IgE: 1 296.33 ± 32.72 μg/L vs. 2 323.33 ± 502.43 μg/L, P < 0.05), so were those of total IgG1 (0.46 ± 0.11 μg/L vs. 0.84 ± 0.11 μg/L, P < 0.05)and ovalbumin-specific IgG1 (0.62 ± 0.11 μg/L vs. 0.86 ± 0.07 μg/L, P < 0.05). Conclusion The fusion protein LTβR-Fc can alleviate symptoms of ovalbumin-induced dermatitis in the mouse model likely by suppressing the LIGHT-HVEM signaling pathway, suggesting that this signaling pathway may serve as a target for the treatment of dermatitis(such as atopic dermatitis).

Pulmonary drug delivery has become a research focus in recent years, while the action between particles and pulmonary macrophages after particles transportation to lung tissue was paid little attention. Therapeutic efficacy can be enhanced by regulating the particles uptake action of pulmonary macrophages according to different diseases. Referring to a lot of articles, the relationship between common diseases and macrophages was reviewed and the influencing factors in the particles uptake of alveolar macrophages were sum-marized. The review laid foundation for the development of preparations and clinical application of pulmonary drug delivery systems.

Chronic spontaneous urticaria (CSU) is a recurrent systemic inflammatory disease. At present, there are few biological indicators that can effectively reflect CSU activity in clinical practice, and some indicators have not been widely used. To identify sensitive and easy-to-check indicators may be helpful for the diagnosis and treatment of as well as prediction of therapeutic efficacy in CSU. This review comprehensively summarizes current clinical application of indicators related to CSU activity, in order to provide a reference for clinical practice and related scientific research.

OBJECTIVETo investigate the role of TRB3 in the inhibitory effect of fenofibrate against the proliferation of glomerular mesangial cell induced by high glucose.

METHODSRat glomerular mesangial cells (MCs) were cultured in the presence of 5.5 mmol/L glucose (normal control), 25 mmol/L glucose (high glucose group), or high glucose along with 10, 50, or 100 µmol/L fenofibrate. Cell counting kit-8 (CCK-8) assay was used to evaluate cell proliferation, and Hoechst 33258 staining was employed to determine chromatin distribution in the MCs. Flow cytometry was performed to analyze the cell cycle changes in different groups. The expressions of TRB3 and P-AKT in different groups were detected using immunocytochemistry and Western blotting.

RESULTSHigh glucose induced obvious proliferation of the MCs (P<0.001), which was significantly inhibited by fenofibrate in a concentration-dependent manner (P<0.001). The MCs exposed to fenofibrate presented with typical apoptotic morphologies and cell cycle arrest at G1/S phase. Low levels of TRB3 expression was detected in the normal control and high glucose groups, whereas in the 3 fenofibrate groups, TRB3 expression increased and P-AKT expression decreased as fenofibrate concentration increased.

CONCLUSIONFenofibrate can promote TRB3 expression in rat MCs. TRB3 causes cell cycle arrest at G1/S phase by inhibiting AKT phosphorylation to result in suppressed proliferation of the MCs.

Animals ; Cell Cycle Checkpoints ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fenofibrate ; pharmacology ; Glucose ; adverse effects ; metabolism ; Mesangial Cells ; cytology ; drug effects ; Phosphorylation ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Signal Transduction

Objective To explore the effect of radiofrequency heating on type Ⅱ collagen expression in a rabbit model of osteoarthritis.Methods Knee osteoarthritis was induced in the right hind legs of 54 male rabbits using modified Hulth modeling.The rabbits were randomly divided into a model group which was not given any special treatment,a Lugua polypeptide group and a radiofrequency hyperthermia group.The Lugua polypeptide group was injected with Lugua polypeptide;the radiofrequency hyperthermia group was treated with radiofrequency irradiation.Six,12 and 18 days after the treatment,the morphological condition of the rats' right femoral medial condyle cartilages were evaluated using modified Mankins scoring and the type Ⅱ collagen content of the cartilage was detected using a quantitative PCR technique.Results At the same time points after treatment,the average Mankins scores were decreased in all the 3 groups,with that of the model group was significantly higher than those of both of the other groups,and the radiofrequency hyperthermia group's average score was significantly better than that of the Lugua polypeptide group.The average type Ⅱ collagen content was significantly increased in all the 3 groups to various extent (the radiofrequency hyperthermia group ＞ Lugua polypeptide group ＞ model group).For the radiofrequency hyperthermia group,the average Mankins score decreased significantly and the average type Ⅱ collagen content increased significantly as the treatment continued.Conclusion Radiofrequency hyperthermia is superior to Lugua polypeptide for treating knee osteoarthritis,at least in rabbits.Its therapeutic effectiveness may be related to a significant increase of type Ⅱ collagen in the cartilage.

Objective To explore the effect of radiofrequency-induced hyperthermia on the morphology of articular cartilage and any changes in serum-1 interleukin-1 (IL-1β) and tumor necrosis factor-alpha (TNF-α) in the process of knee osteoarthritis in rabbits.Methods Fifty-four male rabbits were selected and knee osteoarthritis was introduced to their right hind limbs using the modified Huhh model.They were then randomly divided into a model group,a cervus and cucumis polypeptide (CCP) group and a radiofrequency thermotherapy (RT) group,each of 18.The CCP group was injected with deer melon peptide intramuscularly.The RT group was given radiofrequency hyperthermia treatment.The model group was not provided with any special treatment.On the 7th,13th and 19th day of the treatment,6 rabbits in each group were sacrificed to resect the right medial femoral condyle cartilage.The morphological characteristics of the cartilage were evaluated using the modified Mankins score,while the content of IL-1βand TNF-α in the serum were detected using enzyme-linked immuno-sorbent assays (ELISAs).Results At the same time points,the average Mankins score and the average content of IL-1βand TNF-α in the serum of the model group were significantly higher than in the CCP group,with the values in the latter group significantly higher than in the RT group.In the RT group,the average Mankins score,as well as the IL-1 beta and TNF-alpha levels in the serum,decreased significantly with longer treatment.Conclusion Radiofrequency-induced hyperthermia is superior to deer melon polypeptide in treating knee osteoarthritis,at least in rabbits.Its therapeutic mechanism may be related to the control of serum IL-1 beta and TNF-alpha levels.

BACKGROUND/AIMS: Abdominal pain can be evoked or exacerbated after gastrointestinal cold stimulation in some patients with diarrhea-predominant irritable bowel syndrome (IBS-D), indicating a low temperature-induced sensitization of visceral perception. We investigated the role of vagal transient receptor potential ankyrin 1 (TRPA1, a cold-sensing ion channel) in cold-aggravated visceral mechanonociception in a stress-induced IBS animal model. METHODS: TRPA1 expression was examined in antral biopsies of healthy controls and IBS-D patients. Abdominal symptoms were assessed before and after warm or cold water intake. The visceromotor response (VMR) to colorectal distention (CRD) following intra-antral infusion of cold saline was measured in animals undergoing sham or chronic water avoidance stress. TRPA1 expression, extracellular signal-regulated protein kinase 1/2 (ERK1/2) phosphorylation, and neuronal calcium influx in vagal afferents were assessed. RESULTS: Compared to healthy controls, IBS-D patients displayed elevated antral TRPA1 expression, which was associated with symptom scores after cold (4°C) water intake. Intra-antral infusion of cold saline increased VMR to CRD in naive rats, an effect dependent on vagal afferents. In stressed rats, this effect was greatly enhanced. Functional blockade and gene deletion of TRPA1 abolished the cold effect on visceral nociception. TRPA1 expression in vagal (but not spinal) afferents increased after stress. Moreover, the cold-induced, TRPA1-dependent ERK1/2 activation and calcium influx in nodose neurons were more robust in stressed rats. CONCLUSIONS: Stress-exaggerated visceral mechanonociception after antral cold exposure may involve up-regulation of TRPA1 expression and function on vagal afferents. Our findings reveal a novel mechanism for abnormal gastrointestinal cold sensing in IBS.
Abdominal Pain ; Animals ; Ankyrins ; Biopsy ; Calcium ; Cold Temperature ; Drinking ; Gene Deletion ; Humans ; Irritable Bowel Syndrome ; Models, Animal ; Neurons ; Nociception ; Phosphorylation ; Protein Kinases ; Rats ; Stress, Psychological ; Up-Regulation ; Vagus Nerve ; Visceral Pain ; Water

Cholestatic liver diseases (CLD) are a series of diseases due to impaired bile flow and accumulation of bile acid in the liver and/or systemic circulation caused by immune, genetic, and environmental factors. The pathogenesis of CLD remains unclear and CLD is difficult to treat. As a substitute for human diseases, animal models can provide a platform for exploring the etiology and pathogenesis of the disease and finding appropriate therapeutic targets. This article reviews the current research advances in the animal models of CLD.