Effects of polygonatum polysaccharide on p38 MAPK/N-cadherin in zebrafish with Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To study the effect of polygonatum polysaccharide on zebrafish with Alzheimer disease. METHODS Zebrafish were trained in T maze for 7 d. The 40 zebrafish successfully trained were divided into 4 groups:blank group, model group, positive group and polygonatum polysaccharide group. Model group, positive group and polygonatum polysaccharide group were put in AlCl3100μg·L-1 for 6 d. The positive group was exposed to Huperzine A solution 4μg·L-1, and the polygonatum polysaccharide group was exposed to polygonatum polysaccharide solution 6 g·L-1 for 6 d. The model group was not treated, and the blank group was not treated. Each stage of zebrafish was recorded by video, and the time of each group in the EC region was analyzed. After administration, the brain tissue was taken out and the expression of N-cadherin, P38 and p-P38 protein factors was determined by Western blotting. RESULTS In behavior, the analysis of the time spent in the EC area, the blank group, the positive group and the polygonatum polysac?charide group were compared with the model group, respectively, there were statistically significant differences (P<0.05). At the protein level, compared with the model group, the P38 and p-P38 proteins in the positive group and the polygonatum polysaccharide group were down-regulated, while the N-cadherin protein was up-regulated, with statistical difference (P<0.05). CONCLUSION Polygonatum polysaccharide can improve the learning and memory ability of zebrafish with Alzheimer disease by up regulating the protein level of N-cadherin and hindering P38 phosphorylation.

Intervention of Modified Yupingfeng San Combined with Aspirin on D-dimer， PagT Levels of RSA with Seronegative APS / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of aspirin combined with modified Yupingfeng San on patients with recurrent spontaneous abortion and Seronegative antiphospholipid syndrome. Method:From December 2016 to March 2018, 109 cases of recurrent spontaneous abortion patients with seronegative antiphospholipid syndrome in Affiliated Hospital of Nanjing University of Chinese Medicine were selected as the study objects. According to the random number table, they were divided into three groups:blank group, control group and study group. The blank group was treated with folic acid tablets, 1 pill/time, qd, as placebo, the control group was treated with folic acid tablets, 1 pill/time, qd, Danshen pills, 4 pills/time, tid, and aspirin, 1 pill/time, tid, and the study group was treated with folic acid tablets, 1 pill/time, qd, aspirin, 1 pill/time, tid, and modified Yupingfeng San, 150 mL/time, bid. These patients were all asked for pregnancy preparation for 1 year and treated with aspirin, 1 pill/time, tid, and progesterone pills, 1 pill/time, tid, since the beginning of pregnancy until 13 weeks or abortion. The pregnancy rate, miscarriage rate, coagulation index before and after treatment, traditional Chinese medicine (TCM) symptom score as well as months of abortion were all observed and compared. Result:The coagulation index and TCM symptom score of the study group were significantly lower than those before treatment (PPPPPConclusion:The data proved that the combination of anti-coagulation medicine with either single herb or herbal compound can improve the curative effect of the patients, and traditional Chinese herbal compound can regulate the immune system, reduce the thrombosis and improve the symptoms of TCM, which made the curative efficacy more prominent. However, more samples were needed for further research. Because the time of fetal protection may not change the outcome of pregnancy, the therapy should be maintained at least until the end of the early pregnancy.

miR-140-3p Knockdown Suppresses Cell Proliferation and Fibrogenesis in Hepatic Stellate Cells via PTEN-Mediated AKT/mTOR Signaling

PURPOSE: Liver fibrosis is a major cause of morbidity and mortality and the outcome of various chronic liver diseases. Activation of hepatic stellate cells (HSCs) is the key event in liver fibrosis. Studies have confirmed that miR-140-3p plays a potential regulatory effect on HSC activation. However, whether miR-140-3p mediates the liver fibrosis remains unknown. MATERIALS AND METHODS: Expression of miR-140-3p was detected by real-time quantitative PCR (qPCR). Cell proliferation was measured by MTT, while cell apoptosis rate was determined via flow cytometry. Western blot assay was used to detect the expression of cleaved PARP. The fibrogenic effect was evaluated by expression of α-smooth muscle actin and desmin. Functional experiments were performed in transforming growth factor β1 (TGF-β1)-induced HSC-T6 cells with transfection of anti-miR-140-3p and/or siPTEN. Target binding between miR-140-3p and PTEN was predicted by the TargetScan database and identified using luciferase reporter assay and RNA immunoprecipitation. RESULTS: TGF-β1 induced the activation of HSC-T6 cells, and miR-140-3p expression varied according to HSC-T6 cell activation status. Knockdown of miR-140-3p reduced cell proliferation and the expressions of α-SMA and desmin, as well as increased apoptosis, in TGF-β1-induced HSC-T6 cells, which could be blocked by PTEN silencing. Additionally, inactivation of the AKT/mTOR signaling pathway stimulated by miR-140-3p knockdown was abolished when silencing PTEN expression. PTEN was negatively regulated by miR-140-3p via direct binding in HSC-T6 cells. CONCLUSION: miR-140-3p is an important mediator in HSC-T6 cell activation, and miR-140-3p knockdown suppresses cell proliferation and fibrogenesis in TGF-β1-induced HSC-T6 cells, indicating that miR-140-3p may be a potential novel molecular target for liver fibrosis.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Pioglitazone inhibits cardiac hypertrophy of rats in vitro and in vivo / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effects of pioglitazone on cardiac hypertrophy in vitro and in vivo.</p><p><b>METHODS</b>Angiotensin II was used to establish hypertrophy of cardiac myocytes and pioglitazone was applied to these myocytes in various dosages in vitro. ANP and BNP mRNA expression was evaluated by RT-PCR, and the rate of protein synthesis in CM by 3H-leucine incorporation in cardiac myocytes. Left ventricular hypertrophy was induced by incomplete ligation of abdominal aorta of rats and pioglitazone (20 mg x kg(-1). day(-1)) was administrated one week prior to the operation until 4 weeks after the operation. Cytokines mRNA expression in left ventricle was measured by RT-PCR, left ventricular wall thickness and myocyte diameter were determined by pathological method.</p><p><b>RESULTS</b>Pioglitazone inhibited ANP and BNP mRNA expression and 3H-leucine incorporation in neonatal rat cardiac myocytes induced by angiotensin II in a dose-dependent manner in vitro. Furthermore, pioglitazone reduced the mRNA expression of proinflammatory cytokines, including interleukin-1 beta and cardiotrophin-1, and inhibited the pressure overload-induced increase in the ratio of heart weight to body weight, left ventricular wall thickness and myocyte diameter of rats in vivo.</p><p><b>CONCLUSION</b>Pioglitazone inhibits cardiac hypertrophy of rats in vitro and in vivo, and may play a role in prevention and treatment of cardiovascular diseases characterized by cardiac hypertrophy in future.</p>

Study on Growth and Maturation of Megakaryocyte Progenitors In Vitro in Patients with Chronic Idiopathic Thrombocytopenic Purpura / 中国实验血液学杂志

To investigate the growth and maturation of megakaryocyte progenitors in patients with chronic idiopathic thrombocytopenic purpura (CITP), plasma clot culture and GPIIIa monoclonal antibody and ABC immuno- histochemical kit were used to assay CFU-Meg and BFU-Meg, and the area and diameter of GPIIIa(+) cells were determined by image analyzer in 33 CITP cases. It was found that CFU-Meg and BFU-Meg were 39.27 +/- 21.44 and 5.62 +/- 3.93 per 2 x 10(5) MNC, respectively, in CITP patients, there were no significant differences with those in control group. While the area of GPIIIa(+) cells was (134.90 +/- 6.08) micro m(2) and diameter was (12.89 +/- 3.66) micro m, those were lower than those in control group. In patients with normal number of megakaryocytes on marrow smears, CFU-Meg and BFU-Meg were 19.43 +/- 7.28 and 4.67 +/- 1.53, respectively, the values were lower as compared to control group. The positive correlation was showed between the total megakaryocytic colonies and the number of megakaryocytes on marrow smears, r = 0.6503, and there was no correlation with blood platelet counts and course of disease. The results suggest that there was a maturation disturbance of megakaryocyte progenitor in CITP patients and lower proliferative potential in patients with normal megakaryocyte counts on marrow smears.