To investigate the protective effects and underlying mechanisms of Qin-Zhu-Liang-Xue decoction (QZLX) in atopic dermatitis (AD) and glucocorticoid resistance, we conducted a single-blinded, randomized controlled clinical trial to evaluate the efficacy and safety of this concoction. Network pharmacology analysis was performed and validated through clinical studies. The efficacy, safety, and mechanism of action of QZLX and glucocorticoid receptor (GR) α recombinant protein were assessed in AD mice induced by 2,4-dinitrofluorobenzene (DNFB). Correlation analysis was performed to determine the clinical relevance of GRα. The trial demonstrated that patients who received QZLX showed considerable improvements in their Scoring Atopic Dermatitis (SCORAD) and Dermatology Life Quality Index (DLQI) scores compared with those who received mizolastine at week 4. Network pharmacological analysis identified GRα as a key target for QZLX in AD treatment. QZLX administration increased the serum GRα expression in AD patients, alleviated AD symptoms in mice, decreased inflammatory cytokine expression, and increased GRα expression without affecting liver or kidney function. In addition, GRα recombinant protein improved AD-like skin lesions in DNFB-induced mice. A negative correlation was observed between GRα expression and clinical parameters, including SCORAD, DLQI, and serum IgE levels. QZLX alleviates AD symptoms through the upregulation of GRα and thus presents a novel therapeutic strategy for the prevention of glucocorticoid resistance in AD management.
Dermatitis, Atopic/drug therapy* ; Animals ; Drugs, Chinese Herbal/administration & dosage* ; Humans ; Mice ; Network Pharmacology ; Male ; Female ; Adult ; Receptors, Glucocorticoid/metabolism* ; Disease Models, Animal ; Single-Blind Method ; Middle Aged ; Young Adult

AIM:The paper is to explore a rapid and simple method for the culture of mouse primary thyroid cells.METHODS:Mouse thyroid cells were isolated by enzyme digestion and cultured with improved medium,and their morphology,characteristics and secretory function were observed within 14 d.RESULTS:In the cultures,the active pri-mary cells were obtained from the thyroid tissue after digestion for 25 min;adherent growth was observed on the 2nd day.And secondary follicles appeared from the 5th to 7th day.Over 95%cells were detected with thyroglobulin.The secretion of total triiodothyronine and total thyroxine maintains over 60%in 7 d.The expression levels of specific genes can still maintain more than 50%in 10 d.CONCLUSION:Mouse thyroid primary cells can be rapidly cultured by this method,and the cells can be used for studying thyroid endocrine secretion within 7 d and studying thyroid genes within 10 d.

Alzheimer's disease (AD) is the first degenerative disease of the nervous system, but no drugs have been found to reverse AD progression. Starting from 2 major pathogenesis of AD, namely amyloid beta (Aβ) cascade and Tau protein, this study systematically reviews anti-Aβ or Tau protein AD new drugs that have entered clinical research; this study also expounds their clinical trial findings and mechanisms, and analyzes the reasons for their success or failure to provide a theoretical basis for AD drug exploitation.

Objective:To explore the method of establishing a modified demyelination and myelination regeneration model induced by dicyclohexanone oxalyl dihydrazone (CPZ) in mice with multiple sclerosis (MS), and to analyze the image markers of demyelination and myelination regeneration in mouse MS model.Methods:After the intragastrically administered with sodium carboxymethyl cellulose (CMCNa) for one week, a total of 30 C57BL/6 male mice were randomly divided into the control group ( n=10), the demyelination group ( n=10), and the remyelination group ( n=10). The mice of the control group were immediately performed MR scanning and pathological specimen obtaining; the mice in the demyelination group were administered with intragastrical CPZ-CMCNa once a day for 6 weeks for inducing demyelination, then received MR scanning and specimen obtaining with the same protocols used in control group; the mice in the remyelination group were administered with intragastrical CPZ-CMCNa once a day for six weeks for demyelination, then CPZ was withdrawn and normal diet was given for another four weeks. Then MR scanning and specimen obtaining were performed with the same protocols used in the other two groups. Regions of interest (ROIs) were set at the rostrum of corpus callosum (rCC), the bilateral normal appearing white matters (NAWM) of the rostrum of corpus callosum, and the bilateral cerebral cortex (Cx). The normalized T 2WI (T 2-normalized), fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) values were compared among the three groups by one-way ANOVA. Results:The demyelination and remyelination mice model of MS were successfully established. The T 2-normalized values of rCC in control group, demyelination group and remyelination group were 0.47±0.03, 0.72±0.04, 0.54±0.04, respectively, with statistically significant difference found ( F=90.511, P<0.05). Post-hoc multiple comparisons showed significant differences among those groups ( P<0.05). There was no significant difference of T 2-normalized value in NAWM and Cx among the three groups ( P>0.05). Moreover, there were significant differences in the FA values (0.36±0.04, 0.29±0.03, and 0.32±0.05), the MD values [(0.572±0.015), (0.598±0.034), and (0.626±0.043)×10 -3 mm 2/s], the AD values [(0.79±0.04), (0.77±0.06), and (0.83±0.04)×10 -3 mm 2/s], and the RD values [(0.46±0.02), (0.51±0.03), and (0.53±0.05)×10 -3 mm 2/s] of rCC of the control group, the demyelination group, and the remyelination group (all P<0.05). Significant difference was found in FA values between the demyelination group and the control group ( P<0.05), and in MD values between the remyelination group and the control group ( P<0.05), as well as in AD values between the remyelination group and the demyelination group ( P<0.05). There were also significant differences in RD values between the remyelination group and the control group, and the demyelination group and the control group (all P<0.05). However, no significant difference was found in all diffusion tensor imaging (DTI) metrics of NAWM and Cx among the three groups (all P>0.05). The LFB-eosin staining showed that the myelin sheath of rCC was lost in the demyelination group, and the rCC was partially regenerated and repaired in the remyelination group. Conclusion:The modified CPZ-CMCNa model can selectively induce demyelination and remyelination of rCC, and the changes of demyelination and remyelination of rCC in the modified CPZ-CMCNa model can be quantitatively detected by T 2WI combined with DTI, which might provide related theoretical basis for the study on dynamic changes of MS lesions.

Objective: To investigate the expression of metastasis-associated protein 2 (MTA2) in human bladder cancer tissues and its effect on the malignant biological behaviors of bladder cancer T24 cells, as well as to explore the effect of MTA2 on the progression of bladder cancer. Methods: Sixty-two cases of human bladder cancer tissues and 28 cases of normal bladder tissues (from patients with cystitis, and pathologically confirmed as normal tissue) were collected at People’s Hospital of Hebei Province during December 2012 and December 2014. The expression of MTA2 in bladder cancer tissues and normal bladder tissues was detected by immunohistochemical staining, and the correlation between MTA2 expression and clinicopathological characteristics of patients was also analyzed. The bladder cancer T24 cell line stably expressing MTA2 was constructed. The effects of MTA2 on the proliferation, colony formation, migration and invasion of bladder cancer T24 cells were detected by MTS, clone formation, scratch healing and Transwell assay, respectively. Results: Immunohistochemical staining showed that MTA2 expression was significantly up-regulated in bladder cancer tissues as compared with normal bladder tissues (P<0.01). The high expression of MTA2 in bladder cancer tissues was not related to gender, age and tumor volume (P>0.05), but was associated with higher TNM stage, histological grade, and lymphatic infiltration and metastasis (all P<0.05). After over-expression of MTA2 in bladder cancer T24 cell line, the proliferation activity of the cells was significantly increased (P<0.05), and the colony formation, scratch healing, migration and invasion ability were significantly increased (all P<0.01). Conclusions: MTA2 is up-regulated in human bladder cancer tissues and can promote the proliferation, tumor formation, migration and invasion of T24 cells.

OBJECTIVE: To investigate the effects of glimepiride combined with metformin on glucose and lipid metabolism, islet function and serum miR-126 expression of newly diagnosed type 2 diabetes patients. METHODS: A total of 100 patients with newly diagnosed type 2 diabetes in Nanchuan Hongren Hospital of Chongqing during Jan. 2014-Jan. 2017 were divided into observation group and control group according to random numble table, with 50 cases in each group. Control group was given Metformin hydrochloride sustained-release tablets (Ⅱ) with initial dose of 0. 5 g, once a day, adjusted to 0. 5 g 12 weeks later, twice a day, maximal dose of 1 g at meal or after meal. Observation group was additionally given Glimepiride tablets 2 mg, once a day, at breakfast, on the basis of control group. Both group were treated at lasted for 24 weeks. The levels of blood glucose (FPG, 2 hPG, HbA1c), blood lipid (TC, TG), islet function (FINS, 2 hINS, FCP, 2 hCP, HOMA-IR), serum miR-126 before and after treatment and the occurrence of ADR were observed in 2 groups. RESULTS: Before treatment, there was no statistical significance in the levels of blood glucose, blood lipid, islet function or serum miR-126 expression between 2 groups (P＞0. 05). After treatment, the levels of blood glucose, blood lipid and HOMA-IR in 2 groups were significantly lower than before treatment, and the levels of blood glucose and HOMA-IR in observation group were significantly lower than control group. The levels of FINS, 2 hINS, FCP and 2 hCP, serum miR-126 expression in 2 groups were significantly higher than before treatment, and the observation group was significantly higher than control group, with statistical significance(P＜0. 05). No obvious ADR was found in 2 groups during treatment. CONCLUSIONS: Glimepiride combined with metformin can significantly improve glucose and lipid metabolism, islet function, and regulate serum miR-126 expression without increasing the occurrence of ADR.

Objective To observe the effects of Yishen Tongluo Decoction (YTD) on the renal mRNA expression of transforming growth factor-β1 ( TGF-β1) and collagen Ⅳ ( ColⅣ) in membranous nephropathy ( MN) rats. Methods SD rats were randomly divided into normal group, model group, benazepril group (in the dosage of 10 mg·kg-1·d-1) , YTD group ( in the dosage of 20 g·kg-1·d-1) . The rats in various groups were given intragastric administration of corresponding agents. At the end of the fourth week, 24-hour urinary protein quantity, albumin ( ALB) , total protein ( TP) , triglyceride ( TG) , total cholesterol ( TC) , blood urea nitrogen ( BUN) , and creatinine (Cr) levels were observed. The mRNA expression levels of TGF-β1 and ColⅣ in renal tissue of rats were detected by immunofluorescence method, electron microscopy and real-time polymerase chain reaction (PCR) method. Results In the model group, urinary protein quantity in rats was increased, serum levels of TP and ALB were significantly lowered, serum levels of TC and TG were significantly increased, renal pathological changes were present, and mRNA expression levels of TGF-β1 and ColIV in renal tissue were up-regulated (P<0.05 compared with normal group). Compared with the model group, 24-hour urinary protein quantity, TC and TG levels were significantly lowered, TP and ALB levels were significantly increased, rat renal injury was relieved, mRNA expression levels of TGF-β1 and ColIV in renal tissue were down-regulated in the treatment groups ( P<0.05) . However, the differences between benazepril group and YTD group were insignificant ( P>0.05) . Conclusion The therapeutic mechanism of YTD for MN is probably related with the inhibition of mRNA expression of TGF-β1 and ColⅣin renal tissue.

To explore the effect of PluronicF-127 and Vascular endthelial growth factor(VEGF) delivery system on the survival of the grafted fat, we divided fat harvest under the same condition into 4 groups. One group served as blank control; the other 3 groups served for experiments with respective to DMEM containing 20 ng/ml VEGF; DMEM containing 30% Pluronic F-127; DMEM containing 20 ng/ml VEGF and 30% Pluronic F-127, and then we transplanted the 4 groups of fat, subcutaneously, on the back of 3 groups of BALB/c nude mice (8 mice per group; injecting 4 points per mouse; 0.2 ml per point). At 3 weeks, 6 weeks and 12 weeks, we dissected the fat grafts, measured their weight retention, and put them in histopathologic examination with the use of HE and CD34 staining. And we compared the weight retention and microvessel density (MVD) of each experiment group versus those of control group. The relation between adipose cell and PluronicF-127 was observed through electron microscope. The results reveal that the MVD and weight of pluronicF-127 and VEGF of the experiment groups are significantly greater than those of other groups. The PluronicF-127 and VEGF composite delivery system can significantly improve the blood circulation for fat transplantation, and increase the survival rate of grafted fat.
Adipose Tissue ; cytology ; transplantation ; Animals ; Drug Delivery Systems ; Female ; Graft Survival ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Poloxamer ; administration & dosage ; Random Allocation ; Vascular Endothelial Growth Factor A ; administration & dosage

Objective To investigate the effects of exendin-4 (exenatide) on insulin sensitivity and adipocytokine in high-fat-fed rats. Methods Rats were divided randomly into normal-chow group (NC), high-fat group (HF) and high-fat+exendin treated group (HE). HE rats were given exenatide (2 μg/kg) twice daily for 6 wk. The insulin sensitivity was evaluated by intravenous insulin tolerance test (IVITT). Insulin-stimulated changes in insulin signal transduction, visfatin and adiponectin mRNA expressions as well as their plasma levels were also observed in these rats. Results Plasma free fatty acids (FFA), triglyceride (TG), total cholesterol (TC) levels were significantly reduced after exenatide treatment (in HE rats all P<0.01). And IVITT parameters were also improved in these rats. Insulin-stimulated IRS-1 tyrosine phosphorylation was slightly increased in exenatide-treated rats as compared with HF rats (P<0.05). In addition,plasma visfatin level was significantly reduced in HF and HE groups as compared with controls (P<0.05 and P<0.01). The adiponectin mRNA expression in adipose tissues and circulating adiponectin level were significantly elevated in exenatide-treated rats as compared with untreated rats and controls (P<0.01). Conclusions Chronic exenatide treatment improves insulin resistance in high-fat-fed rats, and the changes of IRS-1 tyrosine phosphorylation and adiponectin may be related to the role of exenatide in elevating insulin sensitivity