Objective To determine the expression level of microRNA-25 in animal models of diabetic nephropathy and human renal tubular epithelial cells (HK-2) cultured in different conditions, and to explore its regulating effect on the fibrosis in diabetic nephropathy. Methods The expression of microRNA-25 was detected by real-time PCR. The downstream target protein of microRNA-25 was verified by bioinformatic prediction, transient transfection of cells and Western blotting. Results MicroRNA-25 was down-regulated in animal models of diabetic nephropathy and HK-2 cells which were cultured in high-glucose medium (P<0. 01). MAP2K4 might be the downstream target protein of microRNA-25. Overexpression of microRNA-25 reduced the protein expression of MAP2K4 and α-SMA (P<0. 01). Conclusion MicroRNA-25 inhibits the fibrosis in diabetic nephropathy by regulating the expression of MAP2K4.

BACKGROUND:Intensity-time (I/T) curve examination is a monitoring technology used for diagnosis of nerve damage,muscle disease and prognostic evaluation using current stimulation and qualitative or quantitative analysis.It also has significance to the diagnosis of lumbar disc herniation (LDH).Whether it can be used in LDH prognostic evaluation is poorly understood.OBJECTIVE:To explore the effectiveness of I/T curve in diagnosis and prognostic evaluation of LDH.METHODS:I/T curves of 113 LDH patients were measured by CX-3 electrodiagnostic equipment,and the results were compared with their unaffected sides and analyzed after physiotherapy.A total of 253 curves were measured,including 107 biceps femoris,101 gastrocnemius and 45 tibialis anterior muscle curves.All patients were sequential treated by traction,medium frequency,ultrashort wave,electric acupuncture as well as infrared radiation following I/T diagnosis,once a day,10 days for a course,with 10 days interval in 3 courses,I/T curves were performed after 3 courses.The therapeutic effect was evaluated by using I/T results combined with clinical symptom and physical signs.RESULTS AND CONCLUSION:The first estimated percentage by I/T curved line were 36.36% (controlled by normal nerve),62.85% (controlled by part of the denervated),0.79% (controlled by the completely denervated) and 63.64% (controlled by total abnormal nerve).After the physiotherapy,the effectiveness of the complete recovery was 92.86% by the normal nerve as well as 58.82% by the abnormal nerve.Above all,the practical value could be concluded from the diagnosis,evaluation and effectiveness of LDH used by I/T curve line.The therapeutic effectiveness controlled by the normal nerve is much better than that controlled by the abnormal.

Objective To observe the immunologic effect of transplantation of MSCs by studying the early and later period of collagen induced arthritis. Methods Rats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture plastic bottle, and the phenotypes were assessed by flow cytometry. We established collagen induced arthritis rats model. MSCs wereinjected from tail veins. We observed the expression of Foxp3 mRNA using RT-PCR, and the level of CD4+CD25+ T cell was tested by flow cytometry. One-way ANOVA and LSD-t test were used for statistical analysis.Results The percentage of CD4+ CD25+ T cells in early and later CIA groups was lower than that of normal control group and treatment groups, which showed statistically significant difference (P＜0.05). The level of CD4+ CD25+ T cell in early MSCs treatment group was higher than the later MSNs treatment group, which showed statistically significant difference (P＜0.05). Compared to the normal group and treatment groups, the expression level of Foxp3 mRNA in the early and later CIA groups was decreased markedly, while the early MSNs treatment group versus the later treatment group showed no statistically significant difference (P＞0.05).The intensity of Foxp3 mRNA in the treatment groups was similar to normal control group. Conclusion In this study, MSCs has shown significant immune-modulatory effects. It up-regulates the level of CD4+ CD25+ T cell in CIA rats, accelerate the expression of Foxp3 mRNA. The early treatment group is more effective than the late treatment group.

Objective To investigate the clinical predictive value of rheumatoid factor (RF), anti-keratin antibodies (AKA), anti-cyclic citrullinated peptide (anti-CCP) antibody and anti-perinuclear factor (APF) in the transition from undifferentiated arthritis (UA) to rheumatoid arthritis (RA), and to analyze the clinical relevant factors. Methods 271 patients with UA who were followed up for a year were enrolled into the investigation. RF was measured by the rate scatting immunity method. APF and AKA were detected by immuniofluorescence method(IFA). Anti-CCP was measured by ELISA. Erythrocyte sedimentation rate (ESR) was analyzed by Wilcoxon method. The duration of morning stiffness, the numbers of swelling and tender joints, tender joints involved and DAS28 score were recorded and analyzed. Results 99% patients who had four-antibody-positive finally convetted from UA to RA. The conversation rate for those who had two or more than two antibody-positive was 83.0% and 65.9% respectively. The sensitivity and specificity of RF and anti-CCP antibody-positive in those who converted from UA to RA was 77.8% and 80.5% respectively. The percentage of polyarticular swelling in antibody-negative, one-antibody-positive, two-antibody-positive, three-antibody- positive and four-antibody-positive was 48%, 57%, 59%, 70% and 70% respectively. Meanwhile, the percentage of multi-small-joint involvement was 71%, 71%, 72%, 76% and 83% respectively. The proportion of elbow involvement in antibody-negative patients was 72%, which was the highest among all joint area involvement. The conversion differences of the group with more than 3 swelling joints or more than 3 small joints involvement ranked the first and second in frequency. Conclusion The combined detection of these autoantibodies could increase the specificity of early diagnosis of RA. The more positive antibodies present, the more likely the concersion form UA to RA. The sensitivity and specificity of RF and anti-CCP-positive is high, so the latter is expected to become one of the diagnostic criteria. The polyarticular swelling and multi-small-joint involvement are valuable in predicting the transformation from UA to RA.

The Hippo signaling pathway is highly conserved in evolution, and participates in the regulation of cell proliferation, differentiation, and tissular dynamic balance, and plays an important role in regulating tissue, organ size, and cell number. Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease and one of the most common causes of end-stage renal disease. Emerging studies have identified the Hippo signaling pathway is closely related to the occurrence and development of ADPKD. The abnormal activity and expression of the main members of the pathway affect the cilia and cell polarity of renal tubular epithelial cells and induce the formation of renal cysts. The review summarizes the potential mechanism of the Hippo pathway in the pathogenesis of ADPKD, the crosstalk with other signaling pathways, and the variances in different species, and discusses the strategies for the treatment of ADPKD based on the Hippo signaling pathway to provide new strategies for the treatment.

Objective To study the inhibitory effect of curcumin on the proliferation,migration and invasion of non-small cell lung cancer cell A549,and to discuss further if it is closely related to the expression of c-Jun N-terminal kinase (JNK) and relative protein p38.Methods A549 cells were cultured by conventional method,and then treated with different concentration of curcumin (10,20,40,80 μmol · L-1).The proliferation,migration and invasion of A549 cells were measured by real-time cellular analysis (RTCA).The expression levels of JNK,p-JNK,p38 and P-p38 were detected by real-time PCR and Western blotting.Results Curcumin showed an antiproliferation effect against A549 cells with IC50 =40 μmol · L-1,and curcumin exhibited obviously inhibitory effect on the migration and invasion of A549 cells.Additionally,compared with control group,curcumin suppressed the expression of JNK and p38 at the gene level,and significantly inhibited the expression of JNK,P-JNK,p38 and p38 (P＜0.05) at the protein level.Conclusion These results demonstrated that curcumin can inhibit the proliferation,migration and invasion of A549 cells via reducing the level of JNK,p38 phosphorylation,and blocking JNK signal transduction pathway.

Objective:To explore the effects of the γ-aminobutyric acid(GABA) neurons and melanin-concentrating hormone (MCH) neurons of the nucleus accumbens (NAc)-lateral hypothalamic area (LHA) neural pathway on the rewarding feeding(palatable food sweat condensed milk) in the obesity rats.Methods:Total 142 male Wistar rats of SPF grade were divided into normal diet (ND) group ( n=68) and high-fat diet induced obesity (DIO) group ( n=74) according to the principle of body mass matching. The rats in the two groups were given normal diet and high-fat diet for 8 weeks. Eight weeks later, 6 DIO rats were randomly selected to observe the nerve projection from GABA neurons in NAc to MCH neurons in LHA by fluorogold retrograde tracing combined fluorescence immunohistochemistry. And the expressions of c-Fos and MCH in LHA after ingestion of sweet condensed milk(rewarding feeding) were observed by fluorescence immunohistochemistry (6 rats in each group). GABA receptor agonist Musimol or GABA receptor antagonist Bicuculine was microinjected into the nucleus of LHA to observe the effect of GABA on rewarding food intake in ND and DIO rats ( n=8 in each group), and the changes of rewarding food intake after blocking MCH signal ( n=8 in each group). SPSS 17.0 was used for statistical analysis, two-way ANOVA and post hoc Bonferroni test were used for comparison among multiple groups, and t-test was used for comparison between two groups. Results:After 8 weeks of high-fat diet modeling, the intake of delicious food in DIO rats was significantly higher than that in ND rats((12.52±2.29) mL, (7.45±1.23) mL, t=4.778, P<0.01) after satiety.The results of fluorogold retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neurons in NAc projected nerve fibers to neurons in LHA, and GABA A receptors in some neurons in LHA coexisted with MCH.The results of NAc-LHA pathway on delicious food intake showed that the interaction between rat group and drug intervention was significant( F=9.869, P<0.01). Simple effect analysis showed that the intake of delicious food after microinjection of Musimol into LHA nucleus of ND rats was significantly lower than that of microinjection normal saline ((4.25±1.38) mL, (7.29±1.49) mL, P<0.01), while the intake of delicious food after injection of Bicuculine was significantly higher than that of microinjection normal saline((10.72±2.11) mL, (7.29±1.49) mL, P<0.05). The intake of delicious food after microinjection of Musimol into LHA nucleus in DIO group was significantly lower than that of microinjection normal saline((3.51±1.77)mL, (13.68±2.95) mL, P<0.01), but there was no significant difference between microinjection Bicuculine and microinjection normal saline ((14.83±3.44) mL, (13.68±2.95) mL, P>0.05). The results of blocking MCH signal on delicious food intake showed that the interaction effect between SNAP-94847 and Bicuculine intervention was not significant ( F=1.468, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=15.880, P<0.01)and the main effect of Bicuculine intervention was significant ( F=6.930, P<0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of ND rats was significantly less than that of injection normal saline((4.78±1.72) mL, (7.63±2.77) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((6.24±2.18) mL, (4.78±1.72) mL, P>0.05). In the DIO rats, the interaction effect between SNAP-94847 and Bicuculine intervention was not significant( F=0.006, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=18.46, P<0.01) and the main effect of Bicuculine intervention was not significant ( F=2.059, P>0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of DIO rats was significantly lower than that of injection normal saline((6.89±2.11) mL, (12.19±4.36) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((8.72±2.26) mL, (6.89±2.11) mL, P>0.05). Conclusion:GABAergic signal in NAc can regulate the expression of MCH in neurons of LHA. In the DIO rats, the sensitivity of MCH neurons in LHA to satiety signal decreases and the hedonic feeding increases.

Objective:To investigate the expression of serum galectin-3 in elderly patients with or without atrial fibrillation and to explore its impact on the prognosis of patients with atrial fibrillation.Methods:In this retrospective study, 100 patients aged 60 years or older treated at the Department of Geriatric Cardiovascular Medicine, Second Xiangyang Hospital, Central South University between March 2018 and September 2018 were enrolled.Based on electrocardiogram and previous history of atrial fibrillation, 73 participants were assigned to the atrial fibrillation group and 27 patients with sinus rhythm and no history of atrial fibrillation during the same period served as the control group.Elisa kits were used to measure the expression of galectin-3 in both groups, and echocardiography was used to measure the size of each cardiac chamber in patients.The Mann-Whitney test was used to compare galectin-3 levels between the two groups, and binary logistic regression analysis was used to determine risk factors for atrial fibrillation in the elderly.Kaplan-Meier survival curves and Cox proportional risk regression were used to analyze survival and the relationship between galectin-3 and prognosis.Results:Serum galectin-3 levels of patients in the atrial fibrillation group were higher than in the control group[(395.13±113.24)ng/L vs.(328.53±89.11)ng/L, t=2.626, P<0.01]. The level of galectin 3 in participants aged ≥ 65 years(n=48)was higher than in those aged<65 years(n=52), (414.01±105.03)ng/L vs.(343.11±106.01)ng/L( t=2.626, P<0.01). The galectin-3 level had a positive correlation with age( r=0.40, P<0.01), duration of atrial fibrillation( r=0.224, P<0.05)and C-reactive protein( r=0.305, P<0.01), but no correlation with N-terminal pro-B-type natriuretic peptide, atrial or ventricular size and the score of CHA2DS2-VASc in patients with atrial fibrillation, .Galectin-3 was a risk factor in patients with atrial fibrillation( P<0.01). Galectin-3 levels did not affect survival(log-rank=0.990)or prognosis( P>0.05)in patients with atrial fibrillation. Conclusions:Galectin-3 levels in elderly atrial fibrillation patients are higher than in people without atrial fibrillation and are positively correlated with age, duration of atrial fibrillation, and C-reactive protein.Galectin-3 is a risk factor for atrial fibrillation in elderly patients.

OBJECTIVETo optimize the dynamic extraction process of salvianolic acids.

METHODSalvianolic acids was selected as index. The effects of extraction temperature, granularity, solvent multiple, circle value and extraction time were studied on the process of dynamical extraction. The orthogonal experiment was employed to investigate the influence of different parameters.

RESULTThe optimal extracting method of salvianolic acids was as follows: temperature was 80 degrees C, granularity was 4 mm, circle value was 30 L x h(-1), solvent multiple was 10 times and extraction time was 150 min.

CONCLUSIONBy comparison to static extraction, dynamic extraction can improve the extraction efficiency, reduce the solvent and energy consumption, as well as lower the burden of post-processing.

Objective To investigate the influence of curcumin and its analogue H8 on glucose and lipid metabolism disorder in db/db mice. Methods The type 2 diabetes mouse model (db/db mice) was intragastrically administrated with curcumin and analogue H8 for 8 weeks.The blood biochemical indexes were measured.The expression of PEPCK and G6Pase mRNA was detected by real-time PCR in liver tissues.The expression of PEPCK and G6Pase protein was detected by Western blotting. Results Curcumin analogue H8 reduced blood glucose and lipids in db/db mice (P<0.01) and improved liver function related enzymes significantly.The levels of PEPCK and G6Pase mRNA in db/db mice were significantly decreased (P<0.01) and the expression levels of PEPCK and G6Pase protein were significantly decreased(P<0.01). Conclusion Curcumin analogue H8 improves the glucose and lipid metabolism disorder in db/db mice,and it is related to inhibiting the expression of PEPCK and G6Pase gene and protein.