Screening of miRNA biomarkers in serum exosomes of patients with thyroid nodules at different iodine levels / 临床检验杂志

Objective To comapre and analyze the differences and commonalities of expression profiles of serum exosomal microRNA between patients with thyroid nodules and healthy persons at different iodine levels,and then provide evidence for screening early diag-nostic markers of thyroid nodules at different iodine levels.Methods The peripheral blood samples from 10 patients with thyroid nod-ules and healthy volunteers at different iodine levels were collected.Their serum iodine levels were measured by the arsenic cerium cat-alytic spectrophotometry.Serum exosomal microRNA were extracted and the expression levels of microRNA were determined by the high-throughput sequencing technology.The differential target genes were predicted and further performed Gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Results Compared with healthy volunteers,there were 6 downreg-ulated miRNAs in the patients with thyroid nodules at different iodine levels,namely miR-324-5p,miR-6511b-3p,miR-9903,miR-550a-3p,miR-5001-3p,and miR-3688-3p.Differentially expressed exosomal microRNA could regulate the MAPK signaling path-way,PI3K-AKT signaling pathway,VEGF signaling pathway,and NF-κB signaling pathway.Conclusion Six differentially expressed microRNAs is identified,which may serve as biological markers for the early diagnosis of thyroid nodules at different iodine levels.

Therapeutic effect of ursodeoxycholic acid-berberine supramolecular nanoparticles on ulcerative colitis based on supramolecular system induced by weak bond / 中国中药杂志

Ulcerative colitis(UC) is a recurrent, intractable inflammatory bowel disease. Coptidis Rhizoma and Bovis Calculus, serving as heat-clearing and toxin-removing drugs, have long been used in the treatment of UC. Berberine(BBR) and ursodeoxycholic acid(UDCA), the main active components of Coptidis Rhizoma and Bovis Calculus, respectively, were employed to obtain UDCA-BBR supramolecular nanoparticles by stimulated co-decocting process for enhancing the therapeutic effect on UC. As revealed by the characterization of supramolecular nanoparticles by field emission scanning electron microscopy(FE-SEM) and dynamic light scattering(DLS), the supramolecular nanoparticles were tetrahedral nanoparticles with an average particle size of 180 nm. The molecular structure was described by ultraviolet spectroscopy, fluorescence spectroscopy, infrared spectroscopy, high-resolution mass spectrometry, and hydrogen-nuclear magnetic resonance(H-NMR) spectroscopy. The results showed that the formation of the supramolecular nano-particle was attributed to the mutual electrostatic attraction and hydrophobic interaction between BBR and UDCA. Additionally, supramolecular nanoparticles were also characterized by sustained release and pH sensitivity. The acute UC model was induced by dextran sulfate sodium(DSS) in mice. It was found that supramolecular nanoparticles could effectively improve body mass reduction and colon shortening in mice with UC(P<0.001) and decrease disease activity index(DAI)(P<0.01). There were statistically significant differences between the supramolecular nanoparticles group and the mechanical mixture group(P<0.001, P<0.05). Enzyme-linked immunosorbent assay(ELISA) was used to detect the serum levels of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6), and the results showed that supramolecular nanoparticles could reduce serum TNF-α and IL-6 levels(P<0.001) and exhibited an obvious difference with the mechanical mixture group(P<0.01, P<0.05). Flow cytometry indicated that supramolecular nanoparticles could reduce the recruitment of neutrophils in the lamina propria of the colon(P<0.05), which was significantly different from the mechanical mixture group(P<0.05). These findings suggested that as compared with the mechanical mixture, the supramolecular nanoparticles could effectively improve the symptoms of acute UC in mice. The study provides a new research idea for the poor absorption of small molecules and the unsatisfactory therapeutic effect of traditional Chinese medicine and lays a foundation for the research on the nano-drug delivery system of traditional Chinese medicine.

Preoperative MRI radiomics models for predicting risk stratification of endometrial cancer / 中国医学影像技术

Objective To observe the value of preoperative MRI radiomics models for predicting risk stratification of endometrial cancer(EC).Methods Data of 219 EC patients who underwent pelvic MR examination before surgery were retrospectively analyzed.The patients were divided into high risk group(n=104)or low risk group(n=115)according to postoperative pathological findings,also assigned into training set(n=153)or test set(n=66)according to examination time and further divided into high or low risk subgroups in each set.ROI was manually sketched on MRI using 3D Slicer,and each 1 130 features were extracted from axial and sagittal fat suppression(FS)T2WI as well as axial and sagittal enhanced FS-T1WI,respectively.Then the least absolute shrinkage and selection operator(LASSO)was used to select a total of 54 merged MRI features,including 12,14,16 and 12 features,respectively.Finally,25 merged LASSO features were reduced dimensionality and selected by reusing LASSO.Extremely randomized trees algorithm was used to construct radiomics models based on each single sequence features,merged MRI features and merged LASSO features,respectively.Receiver operating characteristic curves were drawn,the area under the curve(AUC),the accuracy and F1 score were obtained to evaluate the predicting efficacy of each model.AUC was used to evaluate the predictive efficacy of the models and subjective diagnosis of test set.Results In training set,the accuracy(0.784,0.777),F1 score(0.730,0.731)and AUC(0.835,0.855)of modelmerged MRI and modelmerged LASSO were both higher than those of each single sequence model,while in test set,the sensitivity(0.794,0.882),specificity(0.909,0.969)and AUC(0.904,0.934)of modelmerged MRI and modelmerged LASSO were both higher than those of subjective diagnosis and each single sequence model.The predictive effiency of modelmerged LAsSo was better than that of modelmerged MRI,which was the best model.Conclusion Preoperative MRI radiomics model was effective for predicting risk stratification of endometrial cancer.Modelmerged LASSO had the best performance.

Sagittaria sagittifolia polysaccharides regulates Nrf2/HO-1 to relieve liver injury caused by multiple heavy metals in vivo and in vitro / 中国中药杂志

This study explored whether Sagittaria sagittifolia polysaccharides(SSP) activates the nuclear factor erythroid-2-related factor2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway to protect against liver damage jointly induced by multiple heavy metals. First, based on the proportion of dietary intake of six heavy metals in rice available in Beijing market, a heavy metal mixture was prepared for inducing mouse liver injury and HepG2 cell injury. Forty male Kunming mice were divided into five groups: control group, model group, glutathione positive control group, and low-and high-dose SSP groups, with eight mice in each group. After 30 days of intragastric administration, the liver injury in mice was observed by HE staining. In the in vitro experiment, MTT assay was conducted to detect the effects of SSP at 0.25, 0.5, 1, and 2 mg·mL~(-1) on HepG2 cell survival at different time points. The content of alanine transaminase(ALT) and aspartate aminotransferase(AST) in the 48-h cell culture fluid was measured using micro-plate cultivation method, followed by the detection of the change in reactive oxygen species(ROS) content by flow cytometry. The mRNA expression levels of Nrf2 and HO-1 in cells were determined by RT-PCR, and their protein expression by Western blot. HE staining results showed that compared with the model group, the SSP administration groups exhibited significantly alleviated inflammatory cell infiltration and fatty infiltration in the liver, with better outcomes observed in the high-dose SSP group. In the in vitro MTT assay, compared with the model group, SSP at four concentrations all significantly increased the cell survival rate, decreased the ALT, AST, and ROS content(P<0.05), and down-regulated Nrf2 and HO-1 mRNA and protein expression(P<0.05). SSP significantly improves inflammatory infiltration in the liver tissue of mice exposed to a variety of heavy metals and corrects the liver fat degeneration, which may be related to its regulation of the Nrf2/HO-1 signaling pathway, reduction of ROS, and alleviation of oxidative damage.

Visualization of Literature Information on Postmortem Interval Estimation Indexed by CNKI Database from 1990 to 2020 / 法医学杂志

OBJECTIVES@#To explore the development process of the postmortem interval (PMI) research in China from January 1990 to August 2020, research hotspots in different periods, authors and cooperation between institutions, and to provide a reference for the better development of PMI inference research through the visualization of the literature information of the PMI estimation research indexed in China National Knowledge Infrastructure (CNKI).@*METHODS@#The information visualization analysis software CiteSpace 5.7.R1 was used to carry out big data analysis on hotspots, high-frequency keywords, authors, institutions and other information in the research literature on PMI inference from January 1990 to August 2020 indexed in CNKI.@*RESULTS@#The peak time of publication of PMI was from 2006 to 2010 with 114 articles. In keyword co-occurrence network, the effective hot words were forensic entomology, DNA content analysis and some emerging words such as artificial intelligence and big data. In the cooperation network of institutions, the high-frequency institutions were mainly the scientific research institutions. The author cooperation network showed a trend of co-aggregation and multi-cooperation.@*CONCLUSIONS@#With the development of science and technology, the research on PMI estimation based on traditional methods is mature and novel strategies are emerging. Big data and artificial intelligence combined with forensic science provide new research directions on PMI estimation.

Allium tuberosum alleviates pulmonary inflammation by inhibiting activation of innate lymphoid cells and modulating intestinal microbiota in asthmatic mice / 中西医结合学报

OBJECTIVE@#This study tests whether long-term intake of Allium tuberosum (AT) can alleviate pulmonary inflammation in ovalbumin (OVA)-induced asthmatic mice and evaluates its effect on the intestinal microbiota and innate lymphoid cells (ILCs).@*METHODS@#BALB/c mice were divided into three groups: phosphate buffer saline, OVA and OVA + AT. The asthmatic murine model was established by sensitization and challenge of OVA in the OVA and OVA + AT groups. AT was given to the OVA + AT group by oral gavage from day 0 to day 27. On day 28, mice were sacrificed. Histopathological evaluation of lung tissue was performed using hematoxylin and eosin, and periodic acid-Schiff staining. The levels of IgE in serum, interleukin-5 (IL-5) and IL-13 from bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay. The ILCs from the lung and gut were detected by flow cytometry. 16S ribosomal DNA sequencing was used to analyze the differences in colon microbiota among treatment groups.@*RESULTS@#We found that long-term intake of AT decreased the number of inflammatory cells from BALF, reduced the levels of IL-5 and IL-13 in BALF, and IgE level in serum, and rescued pulmonary histopathology with less mucus secretion in asthmatic mice. 16S ribosomal DNA sequencing results showed that AT strongly affected the colonic bacteria community structure in asthmatic mice, although it had no significant effect on the abundance and diversity of the microbiota. Ruminococcaceae and Desulfovibrionaceae were identified as two biomarkers of the treatment effect of AT. Moreover, AT decreased the numbers of ILCs in both the lung and gut of asthmatic mice.@*CONCLUSION@#The results indicate that AT inhibits pulmonary inflammation, possibly by impeding the activation of ILCs and adjusting the homeostasis of gut microbiota in asthmatic mice.

Identification of COL3A1 variants associated with sporadic thoracic aortic dissection: a case-control study / 医学前沿

Thoracic aortic dissection (TAD) without familial clustering or syndromic features is known as sporadic TAD (STAD). So far, the genetic basis of STAD remains unknown. Whole exome sequencing was performed in 223 STAD patients and 414 healthy controls from the Chinese Han population (N = 637). After population structure and genetic relationship and ancestry analyses, we used the optimal sequence kernel association test to identify the candidate genes or variants of STAD. We found that COL3A1 was significantly relevant to STAD (P = 7.35 × 10

Expression and evaluation of porcine circovirus type 2 capsid protein mediated by recombinant adenoassociated virus 8

Background@#Porcine circovirus type 2 (PCV2) is an important infectious pathogen implicated in porcine circovirus-associated diseases (PCVAD), which has caused significant economic losses in the pig industry worldwide. @*Objectives@#A suitable viral vector-mediated gene transfer platform for the expression of the capsid protein (Cap) is an attractive strategy. @*Methods@#In the present study, a recombinant adeno-associated virus 8 (rAAV8) vector was constructed to encode Cap (Cap-rAAV) in vitro and in vitro after gene transfer. @*Results@#The obtained results showed that Cap could be expressed in HEK293T cells and BABL/c mice. The results of lymphocytes proliferative, as well as immunoglobulin G (IgG) 2a and interferon-γ showed strong cellular immune responses induced by Cap-rAAV. The enzyme-linked immunosorbent assay titers obtained and the IgG1 and interleukin-4 levels showed that humoral immune responses were also induced by Cap-rAAV. Altogether, these results demonstrated that the rAAV8 vaccine Cap-rAAV can induce strong cellular and humoral immune responses, indicating a potential rAAV8 vaccine against PCV2. @*Conclusions@#The injection of rAAV8 encoding PCV2 Cap genes into muscle tissue can ensure long-term, continuous, and systemic expression.

Application of cervix-uterine ITV in volumetric modulated arc therapy for cervical cancer under different bladder filling status / 中华放射肿瘤学杂志

Objective:To investigate the clinical application of cervix-uterine internal target volume (ITV) in volume-modulated arc therapy (VMAT) for cervical cancer under different bladder filling conditions, aiming to obtain more accurate planning target volume (PTV).Methods:Sixty-two patients with cervical cancer receiving radiotherapy rather than surgery were selected and randomly divided into the study ( n=31) and control groups ( n=31). In the study group, individualized ITV, PTV and PTV margin were obtained under three bladder filling status by localization CT scan to compare the VMAT. The target area and organs at risk (OARs) within the target area were statistically compared between two groups. The target missing rate in CBCT, adverse events and short-term efficacy of radiotherapy were analyzed in two groups. Results:There was no significant difference in the volume of target area and OARs in the target area between two groups (both P>0.05). In the study group, the target missing rate and target missing volume were significantly lower compared with those in the control group (both P<0.05). There was no significant difference in the incidence of acute radiation-induced adverse events between two groups ( P>0.05). The 1-, 2-year overall survival and progress-free survival did not significantly differ between two groups (all P>0.05). One patient in the study group had uterine recurrence and 2 cases in the control group. Conclusion:Application of individualized cervix-uterine ITV and PTV in definitive VMAT under different bladder filling conditions can improve the accuracy of target area contouring and improve the local control rate in cervical cancer patients.

Study on the mechanism of Yishen-Qingli granule in treating IgA nephropathy based on network pharmacology / 国际中医中药杂志

Objective:To explore the molecular mechanism of Yishen-Qingli granules in treating IgA nephropathy through network pharmacology. Methods:The relevant components and functional targets of 8 Chinese herbs of Yishen-Qingli granules were screened through the TCMSP. The functional targets were converted into genes in the Uniprot database. Cytoscape 3.6.0 was used to construct the active ingredients-target network; screening IgA nephropathy-related targets through the Human Genome Database (GeneCards), importing drug and disease targets data into Venny2.1 software for intersection, and uploading the shared targets to the STRING platform to build a protein interaction network. With the help of bioinformatics resource database, GO and KEGG enrichment analysis were carried out on the generated results, and the multi-dimensional network of " Yishen-Qingli granule composition target signal pathway" was constructed. Results:69 active ingredients were screened, including ginsenoside, beta-sitosterol, sitosterol, and quercetin. There were 94 Yishen-Qingli granules for IgA nephropathy, including 30 core targets, including JUN, TNF, TP53, AKT1, and IL-6. KEGG enrichment analysis mainly involve three pathways, namely PI3K-Akt pathway, TNF inflammation pathway and MAPK pathway. Conclusions:Yishen-Qingli granules can treat IgA nephropathy in multi-component, multi-target and multi-pathway way. This study initially explored the therapeutic mechanism of Yishen-Qingli granules through network pharmacological analysis, and laid the foundation for further experimental research.

Effects of alprostadil in β-aminopropanitrile induced aortic dissection in a murine model / 中华心血管病杂志

Objective: To investigate the protective role of alprostadil on aortic dissection. Methods: 26 C57BL6 male mice were divided into control group (normal drinking water, n=13) and model group (1 g·kg-1·d-1 BAPN via drinking water, n=13). On day 14, mRNA expression of inflammatory-related genes as well as EP receptor families were detected by RT-PCR (n=6 each) and EP4 protein levels were determined by Western blot (n=7 each). Another 88 mice were divided into 3 groups: control group (n=22), model group (n=33) and treatment group (n=33). The mice in model group and treatment group were applied with BAPN (1 g·kg-1·d-1) via drinking water. The mice in treatment group received additional intraperitoneal injection with alprostadil (80 μg·kg-1·d-1) for 28 days. The mice in the control and model group received equal volume intraperitoneal injection with 0.9% saline respectively. The body weight and systolic blood pressure, the mortality and morbidity were monitored from the beginning until the designed end of the study. On day 28, the mice were sacrificed and aorta were fixed, embedded and sliced, followed by staining with HE and Victoria Blue. The distribution of EP4 was determined by immunohistochemistry in control (n=6) and model group (n=6). Furthermore, the concentration of PGE1 were tested among model (n=3) and treatment group (n=4). EP4 protein expression was determined in model group (n=7) and treatment group (n=6). Results: On day 14, mRNA expression level of MCP-1 ((2.74±1.55) vs. (1.00±0.49),<0.05) and MMP2((1.38±0.42) vs. (1.00±0.27), P<0.05) was significantly upregulated in model group compared with control group. Protein expression of EP4 receptor also increased in aorta in model group compared with control group (1.48±0.51 vs. 1.00±0.19, P<0.05). In the dissection area, the EP4 expression was also enriched compared with non-dissection area, particularly in endothelial cells and inflammatory cells on day 28. BAPN applied in drinking water (model and treatment groups) successfully induced the aortic dissection in mice, some mice died of the rupture. The elastic fibers were fractured, and the infiltrated immune cells were visible in dissected tissue. False lumen was formed. There was no dissection and death in the control group. Compared with control group, the morbidity and mortality rates were significantly increased in the model group (60.6%, 20/33, 30.3%, 10/33) and the treatment group (72.7%, 24/33, 24.2%, 8/33). The mortality and morbidity rates were similar between model and treatment groups. There is no difference in terms of SBP among three groups (P>0.05). Further study showed that after alprostadil injection, the blood concentration of PGE1 was increased in treatment group ((0.540±0.041 vs. 0.436±0.012)μmol/L, P<0.05). Besides, the EP4 receptor expression was downregulated in the treatment group compared to model group (0.60±0.30 vs. 1.00±0.20, P<0.05). Conclusion: EP4 expression is upregulated in BAPN induced aortic dissection mouse model. No protective effects are observed post alprostadil treatment in this model probably due to the reduced expression of EP4.

Ablation of gut microbiota alleviates obesity-induced hepatic steatosis and glucose intolerance by modulating bile acid metabolism in hamsters

Since metabolic process differs between humans and mice, studies were performed in hamsters, which are generally considered to be a more appropriate animal model for studies of obesity-related metabolic disorders. The modulation of gut microbiota, bile acids and the farnesoid X receptor (FXR) axis is correlated with obesity-induced insulin resistance and hepatic steatosis in mice. However, the interactions among the gut microbiota, bile acids and FXR in metabolic disorders remained largely unexplored in hamsters. In the current study, hamsters fed a 60% high-fat diet (HFD) were administered vehicle or an antibiotic cocktail by gavage twice a week for four weeks. Antibiotic treatment alleviated HFD-induced glucose intolerance, hepatic steatosis and inflammation accompanied with decreased hepatic lipogenesis and elevated thermogenesis in subcutaneous white adipose tissue (sWAT). In the livers of antibiotic-treated hamsters, cytochrome P450 family 7 subfamily B member 1 (CYP7B1) in the alternative bile acid synthesis pathway was upregulated, contributing to a more hydrophilic bile acid profile with increased tauro--muricholic acid (TMCA). The intestinal FXR signaling was suppressed but remained unchanged in the liver. This study is of potential translational significance in determining the role of gut microbiota-mediated bile acid metabolism in modulating diet-induced glucose intolerance and hepatic steatosis in the hamster.

Combined use of NLR, V2o and Dmean to predict radiation-induced lung injury in lung cancer patients: an external validation study / 中华放射肿瘤学杂志

Objective To externally validate the accuracy of combined use of neutrophil-lymphocyte ratio (NLR),V20,and Dmean in predicting the incidence of grade Ⅲ or higher radiation-induced lung injury (RILI) in lung cancer patients.Methods A total of 166 lung cancer patients,who participated in the model establishment were selected into the internal validation group,and 85 lung cancer patients who received intensity-modulated radiotherapy in our department between June 2016 and June 2018 were assigned into the external validation group.The incidence rate of grade 3 or higher RILI was statistically compared between the internal and external validation groups.Multivariate logistic analysis was performed for NLR,V20 and Dmean The discrimination degree of the predictive model was evaluated by using ROC curve in combination with NLR,V20 and Dmean The calibration degree of the predictive model was assessed by Hosmer-Lemeshow test.Results The incidence rate of grade 3 or higher RILI in the internal and external validation groups was 23.8％ and 22.9％.Multivariate logistic analysis demonstrated that NLR,V20 and Dmean significantly differed in the internal validation group (P=0.032,0.006 and 0.005).However,only V20 significantly differed in the external validation group (P=0.038).The discrimination and calibration degree of RILI was almost consistent between the internal and external validation groups (both P＞0.05).The area under the curve (AUC) predicted by NLR,V20,Dmean and the combination of three indexes were 0.611,0.646,0.682 and 0.775 in the internal validation group,and 0.544,0.702,0.658 and 0.754 in the external validation group,respectively.The calibration degree in the internal validation group was P=2.797and 0.834,P=2.452 and 0.653 in the external validation group.Conclusion Combined application of NLR,V20 and Dmean can accurately predict the incidence of grade Ⅲ or higher RILI in lung can cancer patients,which has been validated by external dataset.

The mechanisms of medial vascular calcification / 生理学报

Vascular calcification is an active, invertible and highly regulated pathophysiological process, characterized by the deposition of hydroxyapatite crystal in vascular wall. Vascular calcification is classified into two types based on the sites of calcification: intimal atherosclerotic calcification and Mönckeberg's medial calcification. Medial vascular calcification is a pathological phenomenon commonly existed in diabetes, chronic kidney failure and aging. The current review summarizes the mechanisms of medial vascular calcification.

Umbilical cord mesenchymal stem cell modulates immune cell subsets in lupus mice / 中华风湿病学杂志

Objective The purpose of this study is to observe the changes of immune cell subsets in lupus mice after umbilical cord mesenchymal stem cells (UC-MSCs) transplantation. Methods B6.MRL-Faslpr lupus mice were randomly divided into the following three groups: the UC-MSCs treated group, the fibroblast like synoviocytes (FLS) treated group and the untreated group. MSC (1×106) or FLS (1×106) were injected into the tail vein of lupus mice respectively. Four weeks after treatment, the spleen index was calculated. The pathological changes of kidney were assessed by HE staining. The frequencies of immune cell subsets in spleen and macrophage in kidney as well as abdominal cavity were analyzed by flow cytometry. Data were analyzed with t test. Results The spleen index of UC-MSCs treated lupus mice [(79 ±9) mg/10 g] and IgG level [(7.5±1.5) mg/ml] were significantly decreased when compared with FLS treated group [(147±23) mg/10 g, t=2.78, P<0.01] [(17.0 ±2.8) mg/ml, t=3.00, P<0.01] and the untreated group [(156 ±16) mg/10 g, t=4.29, P<0.01] [(16.7 ±1.6) mg/ml,t=4.01, P<0.01]. HE staining also showed that the pathological changes of kidney were alleviated after MSCs transplantation. In addition, the frequency of plasma cells in the untreated group [(2.61 2± 0.318)% vs (0.306±0.017)%, t=7.22, P<0.01] and the FLS treated group [(2.412±0.297)% vs (0.306±0.017)%, t=7.07, P<0.01] were markedly higher than MSCs treatment [(0.306 ±0.017)%]. Moreover, the frequency of CD25+Foxp3+/CD4+Treg in the MSCs treated group [(15.08±0.81)%] was significantly increased compared with the untreated group [(8.02 ±0.47)%, t=7.45, P<0.01] and FLS treated group [(8.80 ±0.23)%, t=7.39, P<0.01]. MSCs treatment resulted in a decrease in CXCR5+PD1+/CD4+Tfh and IFNγ+/CD4+Th1 subset, compared with the untreated group [(14.3±1.5)%vs (31.5±3.3)%, t=5.25, P<0.01] [(1.78±0.27)% vs (5.93±1.56)%, t=2.60, P<0.05] and the FLS treated group [(14.3±1.5)%vs (28.8±2.2)%, t=5.49, P<0.01] [(1.78±0.27)%vs (4.88±0.81)%, t=3.61, P<0.01]. The frequency of macrophage in kidney of the MSCs treated group [(3.52 ±0.37)%] was markedly increased compared with the untreated group[(1.58±0.29)%, t=3.25, P<0.01], while neither the IL4+/CD4+Th2 subset nor the IL17+/CD4+Th17subset and the frequency of macrophage in abdominal cavity showed significant changes in the three groups. Conclusion These findings suggest that the therapeutic effects of MSCs on lupus mice may mediate through increasing the frequency of spleen Treg and renal macrophage and decreasing the frequency of Tfh, Th1 and plasma cells.

Analysis of Clinical Application of Warfarin in Our Hospital / 中国药房

OBJECTIVE:To investigate the clinical application of warfarin in inpatients of our hospital,and provide reference for standardized application in clinic. METHODS:Totally 267 inpatients records used warfarin in our hospital from Jan. 2013 to Jun. 2014 were investigated to retrospectively analyze the indications,dosage,international normalized ratio(INR)monitoring and bleeding complications of warfarin. RESULTS:Warfarin was mainly used for patients with deep vein thrombosis,cerebral infarc-tion,rheumatic heart disease and pulmonary embolism in our hospital;the dosage of inpatients was in the range of 0.625-6.875 mg with average daily dosage of (2.15 ± 0.72) mg;the INR lower than 1.8 accounted for 52.8%,the INR in the range of 1.8-3.0 (reaching the standard) accounted for 31.8% and INR higher than 3.0 accounted for 15.4%;incidence of adverse reactions was 5.6%,mainly subcutaneous blood spots (petechiae) and urine occult blood,and incidence of severe hemorrhage (intracerebral hemorrhage) was 0.7%. CONCLUSIONS:At present,the clinical application of warfarin is still deficient,and qualified rate of INR is still low. It's important to actively carry out medication intervention and clinical pharmaceutical care.

Dynamic alteration of microRNA in high phosphorus induced calcification of vascular smooth muscle cell / 北京大学学报(医学版)

Objective:To study the change of microRNA during the early stage of high phosphorus in-duced vascular smooth muscle cell (VSMC)calcification and its related mechanism.Methods:The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol /L)for 7 d.The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content,real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules.Based on the cell calcification model,micro-RNA microarray array was applied to screen the profiles of microRNA expression in VSMC following high Pi stimulation for different periods (0,3 and 12 h).The array data were analyzed by TAMtool to explore the activated signaling pathway.Results:The calcium content of A7r5 cells induced by high Pi was in-creased 9.6 times high as cells without Pi treatment (P <0.05 ).VSMC contractile phenotype genes (SM-αactin,SM22)were down-regulated (P <0.05 ),while calcification-related genes (BMP2, MSX2,Runx2)were up-regulated (P <0.05)in VSMC stimulated by high Pi.The calcium nodules were obviously formed in cells after 7 d high Pi treatment.In microarray experiment,680 individual mi-croRNAs were detected in high Pi-treated VSMCs at different time points (0,3 and 12 h).Among these genes,miR-183,miR-664 and miR-9 * were increased whereas miR-542-5P,let-7f and miR-29a were decreased in time-dependent manners.Twenty-six kinds of signaling pathways,including cell apoptosis, differentiation and proliferation,were significantly activated.All these activated pathways were associated with calcification.Conclusion:This study implies that microRNA changed in high Pi-induced VSMCs may involve in the process of calcification.

Efficacy of Liraglutide for Obesity or Overweight:A Systematic Review / 医药导报

Objective To evaluate the effectiveness and safety of liraglutide for obesity or overweight. Methods Random controlled trials of liraglutide for obesity or overweight were gathered from MEDLINE, EMbase, CENTRAL, CNKI, VIP and WanFang. We screened the retrieved studies according to the inclusion and exclusion criteria, evaluated the quality of included studies, and then performed meta-analyses with The Cochrane Collaboration's Revman 5. 3. 0 software. Results Twelve randomized controlled trials of liraglutide for obesity or overweight were included, in which 11 trials were written in English and one trial in Chinese. The results of meta-analyses showed that the body weight was significantly reduced in the liraglutide group than in placebo, insulin, exenatide and glimepiride groups [RR=-0. 91,95%CI(-1. 01,-0. 81),P<0. 000 01;RR=-2. 88,95%CI(-3. 37,-2. 39),P<0. 000 01; RR=-1. 12,95%CI(-1. 32,-0. 92),P<0. 000 01; RR=0. 45,95%CI(-0. 62,-0. 27),P<0. 000 01]. Moreover, liraglutide had significant effect in decreasing HbA1c and systolic blood pressure of patients with obesity or overweight. Conclusion Liraglutide is effective for controlling body weight of patients with obesity or overweight. But its long-term efficacy still needs to be confirmed by performing more RCTs with high quality, large sample and long term follow-up.

Umbilical cord mesenchymal stem cell transplantation modulated Th17/Treg balance and delayed lupus progression in MRL/lpr mice / 中华风湿病学杂志

Objective To explore the preventive effect of early umbilical cord mesenchymal stem cells (UC-MSCs) transplantation on MRL/lpr mice and the underly mechanisms.Methods Fourteen 10-week-old MRL/lpr mice were labeled and numbered.They were randomly divided into 2 groups by using random number table and injected with 1 ×106 UC-MSCs or PBS via tail vein respectively.Proteinuria was measured with Bradford method every 4 weeks.All mice were sacrificed at the age of 28 weeks, with the level of serum antidsDNA antibody and IL-17 detected by enzyme linked immunosorbent assay (ELISA).Splenic Th17 cells, as well as regulatory T cells (Treg) were examined by flow cytometry.Data were analyzed with t test and Pearson's correlation test.Results The onset of proteinuria was delayed for 4 weeks in UC-MSC-treated group compared with that in the control group.At the age of 28 weeks, the 24 hour proteinuria [(1.78±0.17) mg vs (4.77±0.98)mg, t=2.99, P＜0.05] and the spleen weight [(0.149±0.009) g vs (0.273±0.052) g, t=2.33, P＜0.05] in UC-MSCtreated group were significantly lower than those in the control group.There was also a trend of the decline of serum anti-dsDNA antibody and IL-17 level after UC-MSCs transplantation.Compared with those in the control group, both the percentage and the absolute number of Th17 cells were significantly decreased in UC-MSC-treated group [(0.90±0.19)％ vs (2.81±0.50)％, t=3.54, P＜0.01 and (3.7±0.8)×105 vs (19.3±3.7)×105, t=4.12,P＜0.01].Meanwhile, the percentage of Treg elevated after UC-MSCs treatment.The ratio of Th17/Treg was significantly lower in UC-MSC-treated group than that in the control group (0.11±0.03 vs 0.50±0.09, t=4.23,P＜0.01).Both the ratio of Th17/Treg (r=0.73, P＜0.01;r=0.59, P＜0.05) and serum IL-17 level (r=0.78, P＜0.01;r=0.56, P＜0.05) was positively correlated with the level of 24 hour proteinuria and anti-dsDNA antibody respectively in MRL/lpr mice.Conclusion Early UC-MSCs transplantation helps to delay disease onset and ameliorate disease progression in MRL/lpr mice, which may act through the modulation of Th17/Treg balance.

Effects of bone marrow mesenchymal stem cells on the bone marrow cells-induced lupus like presentations / 中华风湿病学杂志

Objective Whether the bone marrow cells (BMC) derived from systemic lupus erythematosus (SLE) could transmit autoimmune disease was studied for the purpose of clarifying the role of BMC in SLE pathogenesis.The effects of bone marrow mesenchymal stem cells (MSC) from SLE and control mice on the SLE BMC-induced symptoms were compared to elucidate the role of MSC in SLE.Methods Six-week-old B6.MRL-Fas mice were randomly divided into 3 groups.One group was transplanted with BMC from the 30-week-old B6.MRL-Faslg mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Faslpr mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Faslg mice and bone marrow MSC from the age-matched C57BL/6 mice.Before transplantation,the recipient mice received irradiation by an X-ray source.The levels of serum antinuclear antibody (ANA) and proteinuria were measured with enzyme linked immunosorbent assay (ELISA) and Bradford method every 4 weeks,respectively.The survival rate was recorded.All mice were sacrificed 18 weeks later.Splenic plasma cells,Th1,Th2 and Th17 cells were measured by flow cytometry.Statistical analyses were performed using the independent t test and ANOVA.Results Eight weeks after transplantation,ANA was positive in all the recipient mice.However,there was no significant difference between the three groups (P＞0.05).No proteinuria was observed in all the recipient mice.The mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Fasr mice showed an elevated trend of the percentages of splenic plasma cells,Th1,Th2 and Th17 cells compared with the other two groups,plasma cells [(1.05±0.16)％,(0.58±0.11)％,t=2.53,P＞0.05;(1.05±0.16)％,(0.71±0.18)％,t=1.45,P＞0.05],Th1 cells [(6.6±2.2)％,(5.7±1.0)％,t=0.38,P＞0.05;(6.6±2.2)％,(4.0±1.7)％,t=0.96,P＞0.05],Th2 cells [(3.3±0.4)％,(2.1±0.6)％,t=1.76,P＞0.05;(3.3±0.4)％,(2.2±0.6)％,t=1.51,P＞0.05],Th17 cells [(2.30±0.71)％,(1.31±0.31)％,t=1.27,P＞0.05;(2.30±0.71)％,(1.12±0.27)％,t=1.67,P＞0.05].However,there was no significant difference between the groups.The survival rate of the three groups was 43％,43％ and 80％ respectively.And the survival rate of the mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched C57BL/6 mice was significantly higher than those of the other groups.Conclusion Our results indicate that BMC from SLE can transmit autoimmune disease.The bone marrow MSC can not prevent lupus-like presentations induced by BMC from SLE.Transplantation of bone marrow MSC from C57BL/6 mice can significantly elevate the survival rate.