Research of Influence on Inflammation of Xieheyin on PCOS-IR Mice Based on Intestinal Mucosal Barrier and TLR4/NF-κB/NLRP3 Pathway / 中国实验方剂学杂志

Objective:To investigate the possible mechanism of Xieheyin in alleviating obese polycystic ovary syndrome with insulin resistance（PCOS-IR）and reducing inflammatory response. Method:Ten of sixty SPF femlae C57BL/6J mice were randomly selected as the normal group，and the rest mice were given letrozole 0.002 g·kg^-1 combined with fecal suspension 2 g·kg^-1 for 28 consecutive days to establish model of PCOS-IR.The mice that were successfully modeled were randomized into the model group，metformin group（0.25 g·kg^-1），and low（10 g·kg^-1），medium（20 g·kg^-1），and high-dose（40 g·kg^-1）Xieheyin groups，and administered with the corresponding drugs by gavage，once a day，for four consecutive weeks. Except the normal control group， the mice in the other groups were continuously given fecal suspension combined with letrozole solution to maintain the model during the treatment. The mice were weighed once a week.Levels of fasting blood glucose （FBG） were detected by blood glucose test strips.And enzyme-linked immunosorbent assay （ELISA） method was used to detect serum testosterone（T），follicle stimulating hormone（FSH），luteinizing hormone（LH），fasting insulin（FINS）level，and LH/FSH and Homeostasis model assesment of insulin resistance （HOMA-IR） were calculated．The uterus and ovaries were weighed and fixed.Hematoxylin-eosin（HE）staining was used to observe ovarian tissue pathology morphology. Western blot was used to detect the expression levels of tight junction key molecular zonula occludens 1（ZO-1），occludin in colon tissues，and the expression levels of Toll-like receptor 4/nuclear factor kappa B/Nod-like receptor protein 3（TLR4/NF-<italic>κ</italic>B/NLRP3）signaling pathway and inflammation associated proteins cysteinyl aspartate specific proteinase-1（Caspase-1） and interleukin-1<italic>β</italic>（IL-1<italic>β</italic>） in colon tissues. Result:Compared with normal control group，the body weight of mice in the model control group increased significantly（<italic>P</italic><0.01）. Serum FINS，FBG，HOMA-IR，T，LH/FSH were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The uterine organ ratio were decreased significantly（<italic>P</italic><0.01），while the ovarian organ ratio were significantly increased（<italic>P</italic><0.01）. The number of atresia follicles and cystic dilatation follicles increased significantly，and the number of corpus luteum significantly decreased，the thickness of follicular granulosa cells also decreased，while the white membrane thickness of the ovary increased. Tight junction related ZO-1，occludin proteins in colon tissues were all decreased（<italic>P</italic><0.01）.The relative expression levels of inflammation-related protein IL-1<italic>β</italic>，Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein signaling pathway were significantly increased（<italic>P</italic><0.05）.Compared with model control group， the body weight of mice in the low，middle and high dose Xieheyin group decreased significantly（<italic>P</italic><0.01）. The serum T，LH/FSH，FINS，FBG，HOMA-IR were significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The uterine organ ratio were increased（<italic>P</italic><0.05），while the ovarian organ ratio were decreased（<italic>P</italic><0.05）. The number of cystic follicles decreased and corpus luteum increased，the thickness of follicular granulosa cells increased and be arranged normally，while the white membrane thickness of the ovary increased slightly. The expressions of ZO-1，occludin proteins were increased（<italic>P</italic><0.01）. The expression levels of IL-1<italic>β</italic>，Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein in the high dose group were significantly decreased（<italic>P</italic><0.01）. Conclusion:Xieheyin could activate intestinal TLR4/NF-<italic>κ</italic>B/NLRP3 signaling pathway，inhibit pro-inflammatory factor secretion，improve obesity and IR，which was correlated with rebuilding intestinal mucosal barrier and inhibiting intestinal inflammation．

Value of fasting plasma glucose to screen gestational diabetes mellitus before the 24th gestational week in women with different pre-pregnancy body mass index / 中华医学杂志(英文版)

BACKGROUND@#Gestational diabetes mellitus (GDM) is usually diagnosed between 24th and 28th gestational week using the 75-g oral glucose tolerance test (OGTT). It is difficult to predict GDM before 24th gestational week because fast plasma glucose (FPG) decreases as the gestational age increases. It is controversial that if FPG ≥5.1 mmol/L before 24th gestational week should be intervened or not. The aim of this study was to evaluate the value of FPG to screen GDM before 24th gestational week in women with different pre-pregnancy body mass index (BMI).@*METHODS@#This was a multi-region retrospective cohort study in China. Women who had a singleton live birth between June 20, 2013 and November 30, 2014, resided in Beijing, Guangzhou and Chengdu, and received prenatal care in 21 selected hospitals, were included in this study. Pre-pregnancy BMI, FPG before the 24th gestational week, and one-step GDM screening with 75 g-OGTT at the 24th to 28th gestational weeks were extracted from medical charts and analyzed. The pregnant women were classified into four groups based on pre-pregnancy BMI: Group A (underweight, BMI < 18.5 kg/m), Group B (normal, BMI 18.5-23.9 kg/m), Group C (overweight, BMI 24.0-27.9 kg/m) and Group D (obesity, BMI ≥28.0 kg/m). The trend of FPG before 24th week of gestation was described, and the sensitivity and specificity of using FPG before the 24th gestational week to diagnose GDM among different pre-pregnancy BMI groups were reported. Differences in the means between groups were evaluated using independent sample t-test and analysis of variance. Pearson Chi-square test was used for categorical variables.@*RESULTS@#The prevalence of GDM was 20.0% (6806/34,087) in the study population. FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. FPG was higher in women with higher pre-pregnancy BMI. FPG before the 24th gestational week and pre-pregnancy BMI could be used to predict GDM. The incidence of GDM in women with FPG ≥5.10 mmol/L in the 19th to 24th gestational weeks and pre-pregnancy overweight or obesity was significantly higher than that in women with FPG ≥5.10 mmol/L and pre-pregnancy BMI <24.0 kg/m (78.5% [62/79] vs. 52.9% [64/121], χ = 13.425, P < 0.001).@*CONCLUSIONS@#FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. Pre-pregnancy overweight or obesity was associated with an increased FPG value before the 24th gestational week. FPG ≥5.10 mmol/L between 19 and 24 gestational weeks should be treated as GDM in women with pre-pregnancy overweight and obesity.

Clinical Prognostic Factors in 86 Chinese Patients with Primary Myelodysplastic Syndromes and Trisomy 8: A Single Institution Experience

PURPOSE: The objective was to determine the characteristics and prognostic factors of 86 Chinese patients with trisomy 8 aberrations and compare the prognostic value of International Prognostic System (IPSS) and Revised IPSS (IPSS-R) in this cohort. MATERIALS AND METHODS: A total of 86 cases diagnosed with primary myelodysplastic syndromes (MDS) with isolated tr8 or with tr8 and other additional cytogenetic aberrations diagnosed and treated at the Union Hospital, Tongji Medical College of Huazhong University of Science and Technology between July 2002 and March 2013 were reviewed. RESULTS: The median survival of the entire group was 23.0 months, and acute myeloid leukemia (AML) developed in 43% (37/86) patients within the follow up time. The univariate analysis revealed that overall survival (OS) was correlated with age, thrombocytopenia, absolute neutrophil count, marrow blasts, cytogenetic status and red blood cell transfusion at diagnosis, and the multivariate analysis revealed that age, marrow blasts, cytogenetic status and transfusion dependence were independent parameters for the OS. The cytogenetic complexity and marrow blasts had the strongest impact on the AML transformation by multivariate analysis. Comparing the two prognostic systems, both two systems could successfully discriminate risk groups for survival. IPSS-R was more refined than IPSS for predicting OS, but had no advantage in predicting the risk of AML development. CONCLUSION: This study confirmed the influence of clinical factors on the prognosis of 86 Chinese MDS patients with trisomy 8. In addition, IPSS-R can further refine prognostic discrimination in the IPSS risk categories.

Expression of autophagy related gene BECLIN-1 and number of autophagic vacuoles in bone marrow mononuclear cells from 40 myelodysplastic syndromes patients and their significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to detect the expression level of autophagy related gene BECLIN-1 and the number of autophagic vacuoles in bone marrow mononuclear cells (BMMNC) from myelodysplastic syndrome(MDS) patients and to explore their difference in different stage of MDS and relationship between their difference and disease characteristics.</p><p><b>METHODS</b>The BMMNC from 9 normal controls, 19 cases of low-risk MDS, 14 cases of high-risk MDS and 7 cases of MDS-transformed AML were collected. The expression level of BECLIN-1 was detected by real time PCR (RT-PCR) and the amount of autophagic vacuoles was counted by transmission electron microscopy.</p><p><b>RESULTS</b>The expression level of BECLIN-1 in BMMNC from patients with low-risk group was obviously higher than that in BMMNC from normal controls; the expression level of BECLIN-1 in BMMNC from patients of hgh risk group was higher than that in BMMNC of normal group, but there was no statistical significance (P > 0.05); the expression level of BECLIN-1 in BMMNC from patients with MDS-transformed AML group was significanly lower than that in BMMNC of normal group (P < 0.05). Transinission electron microscopy showed that the amount of autophagic vacuoles in BMMNC from patients with low-risk and high-risk MDS groups was more than that in normal control, but there was no stetistcal significance (P > 0.05), while the amount of autopuagic vecuoles in BMMNC from patients of MDS-transformed AML group was significantly less (P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of BECLIN-1 and the number of autophagic vacuoles in BMMNC from patients with MDS progression and patients with MDS-transformed AML are gradually declining. The autophagy may be associated with disease progression.</p>

Effect of flow shear stress gradient on the arrangement and shape of endothelial cells / 医用生物力学

Objective The effect of different flow shear stress gradient on the changes of arrangement and shape of endothelial cells was evaluated in order to investigate the effect of shear stress gradient on ECs morphology and function. Method A flow chamber system with gradient shear stress was established, in which the range of shear stress is from 15 dyn/cm2 to 6.6 dyn/cm2(1 dyn=10-5 N), and the shear stress gradient is 1.5 dyn/cm2 and 3 dyn/cm2 respectively. After ECs were subjected to the gradient shear stress for 6 hours, cell angle, cell width length ratio, as well as cell shape index of ECs under the different shear stress gradient were examined. Results The cell angles of ECs were straggling under both 1.5 dyn/cm2 and 3 dyn/cm2 shear stress gradient. The cell width length ratio and cell shape index of ECs were decreased under 1.5 dyn/cm2 shear stress gradient compared with that of 3 dyn/cm2 shear stress gradient. Conclusions The ECs show random orientation under the different shear stress gradient. The ECs are trending to stretch and elongate shape under smaller shear stress gradient, and to approach cycloid under larger shear stress gradient.

Effect of glycyrrhetinic acid on invasive capacity of leukemic cells and activity of gelatinase / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of glycyrrhetinic acid (GA) on the invasive capacity of leukemic cells and the activity of intracellular gelatinase.</p><p><b>METHODS</b>The effect of GA, in different concentrations, on the proliferation of cultured K562 and HL-60 leukemic cells in vitro was determined by MTT assay; that on cell invasive capacity was tested by Transwell cubicle matrigel invasion assay; and that on the activity of gelatinase in cells was detected by gelatin zymography.</p><p><b>RESULTS</b>GA showed significant inhibitory effect on the proliferation of K562 and HL-60 leukemic cells; it inhibited the invasive capacity of cells in concentration-dependent manner; and significantly down-regulated the activity of gelatinase A and B in cells.</p><p><b>CONCLUSIONS</b>GA can inhibit invasive capacity of K562 and HL-60 leukemia cells by way of suppressing the activity of gelatinase A and B. This study provides an experimental evidence for preventing extra-medullary infiltration of leukemic cells.</p>

Resveratrol inhibits the secretion of vascular endothelial growth factor and subsequent proliferation in human leukemia U937 cells / 华中科技大学学报（医学）（英德文版）

This study examined the effect of resveratrol on the secretion of vascular endothelial growth factor (VEGF) and subsequent proliferation of human leukemia U937 cells, and explored the mechanisms involved. Human leukemia U937 cells were treated with resveratrol of different concentrations (12.5-200 micromol/L) for different time lengths (12-48 h). The proliferation of the U937 leukemic cells was determined by MTT assay. Apoptosis was observed by Annexin-V-FIFC/PI double staining and flow cytometry (FCM). Cells cycle was analyzed by PI staining and FCM. The content of VEGF was determined by ELISA. Human umbilical vein endothelial cells were examined for vasoformation in vitro after exposures to resveratrol of various concentrations. The results showed that resveratrol inhibited the proliferation of U937 leukemia cells in a dose-and time-dependent manner. Resveratrol induced apoptosis and S-phase cell cycle arrest in human leukemic U937 cells. Resveratrol inhibited the secretion of VEGF in U937 cells. Resveratrol inhibited the vasoformation of human vein endothelial cells in a dose-dependent manner. It was concluded that resveratrol could down-regulate the secretion of VEGF, induce apoptosis and suppress the proliferation of U937 cells.

Experimental study on anticancer effect of curcumin on Raji cells in vitro / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the anticancer effect and mechanism of curcumin on Raji cells in vitro and compared the cytotoxicities of curcumin on Raji cells and normal human peripheral blood mononuclear cell (PBMC).</p><p><b>METHODS</b>The effects of curcumin on proliferation of Raji cells and human PBMC were tested by MTT assay, its effects on apoptosis of them were determined by Annexin-V/PI double-labeled cytometry and TUNEL, and its effects on DNA distribution in Raji cells was studied by PI single labeled cytometry.</p><p><b>RESULTS</b>Curcumin showed marked inhibition on proliferation of Raji cell, could induce Raji cell apoptosis in time- and dose-dependent manner. After curcumin treatment, the cell cycle of Raji cells was blocked in G0/G1 and G2/M phase and those in the S phase decreased proportionally. But curcumin showed no significant effect on inhibiting proliferation or inducing apoptosis on human PBMC.</p><p><b>CONCLUSION</b>Curcumin could regulate the cell cycle of Raji cells and induce its apoptosis, so as to inhibit its proliferation, but with no significant cytotoxicity on human PBMC. It selectively affects the tumor cell.</p>