OBJECTIVETo explore the effect of infusing G-CSF mobilized recipient spleen cells at different time after haploidentical stem cell transplantation(HSCT) on graft-versus-host disease (GVHD) in mice and its possible mechanism.

METHODSForty mice after HSCT were randomly divided into 4 groups (n=10): GVHD positive control group (control group), 1st d recipient cell infusion group after transplantation (+1 d group), 4th d recipient cell infusion group after transplantation(+4 d group), 7th d recipient cell infusion group after transplantation(+7 d group). The mice in control group were injected the normal saline of same equivalent with experimental group which were given the same amount of G-CSF-mobilized recipient spleen cells. The general manifestation and pathological change of GVHD were observed. The expression changes of CD3CD4, CD3CD8cell subsets and FasL in peripheral blood were detected by flow cytometry.

RESULTSThe incidence of GVHD was significantly decreased in +4 d group and the median survival time was longer than 60 days, which was significantly higher than that of control group (24 d), +1 d group (21 d), +7 d group (28 d). (P<0.01, P<0.01, P<0.01). The Fasl expression of peripheral blood T lymphocytes in +4 d group were significantly lower than that in the other 3 groups(P<0.05).

CONCLUSIONThe +4 d infusion of G-CSF mobilized recipient spleen cells on 4th day after haploidentical HSC transplantation can inhibit the expression of FasL in donor T lymphocytes, and significantly reduce the incidence of GVHD.

OBJECTIVETo investigate the effects of microvesicles(MV) isolated from human peripheral blood hematopoietic stem cells(PB-HSC) on immune regulation and hematopoiesis.

METHODSPB-HSCs were separated by density-gradient centrifugation and cultrued. The supernatants of PB-HSC at 48 h were harvested for isolation and purification of MV by using ultracentrifugation. The electron microscopy was used to observe the morphology of MV. The protein level in MV was quantified through bicinchoninic acid(BCA) protein assay. Flow cytometry was used to detect the immunophenotype of MV. Human peripheral blood mononuclear cells(PB-MNC) were isolated from healthy donor and treated with isolated MV. After being co-cultured for 12 h, confocal microscopy was used to observe the action mode of MV on PB-MNC. After being co-cultured for 48 h, the levels of IL-2, IL-6, IL-8, IL-10, IFN-γ and TNF-α were detected by ELISA. Flow cytometry was used to detect the changes of T cell subsets and the activation of T cell subsets as well as intracellular cytokine staining after co-culture for 48 h. The methylcellulose was used to assess the hematopoiesis-supportive function of MV as well as co-cultured supernatants.

RESULTSThe eletron microscopy revealed that MV were elliptical membrane vesicles. The protein amount in MV ranges from 29 to 110 µg. Flow cytometry showed that MV expressed mix markers on the surface, especially highly expressed MV specific marker CD63(85.86%) and hematopoietic stem cell marker CD34(33.52%). After being co-cultured for 12 h, confocal microscopy showed that MV were merged with PB-MNC. After being co-cultured for 48 h, ELISA showed that the secretion of cytokines IL-6,IL-8, IL-10 as well as TNF-α was increased while the level of IL-2 and IFN-γ was not changed much. The results of flow cytometry showed that there was no significant change in T cell subsets and T cell activation. Staining of intracellular factor showed that IL-8 was increased significantly in CD11ccells. The colony-forming experiments revealed that MV and the co-cultured supernatants could facilitate the colony formation.

CONCLUSIONMV isolated from PB-HSC have immune-regulatery function and can prornote hematopoiesis.

In order to obtain the characteristics and incidence of adverse reactions of Shuxuening injection (Xingxue), the design method of a multi-center, large sample intensive monitoring in the hospitals was adopted. The hospitalized patients with use of Shuxuening injection from 27 medical institutions were enrolled as the research subjects to monitor their entire process of treatment cycle. The main content of monitoring included the patients' general information, diagnostic information, medication information, and adverse event information. A total of 30 209 patients with Shuxuening injection were enrolled; adverse reactions occurred in 34 cases, with an incidence of 0.113%, which belonged to the rare adverse reactions. Adverse reactions were characterized by headache, dizziness, pruritus, palpitations, nausea, et al. All the above results showed that Xingxue Shuxuening injection had high safety in clinical application.

OBJECTIVETo explore the hemodynamic changes in cynomolgus monkeys with mild carotid atherosclerotic (CAS) plaques after therapy with pushing manipulation on Qiaogong acupoint (MPQ).

METHODSNine cynomolgus monkeys were equally randomized into MPQ group, mild CAS model group and blank control group. Mild CAS models were established in the monkeys in MPQ and model groups, and the monkeys in MPQ group received treatment with MPQ intervention after the modeling. The conditions of the carotid artery and the hemodynamic changes in the 3 groups were evaluated after the treatment.

RESULTSFormation of CAS plaques was detected in monkeys in both MPQ and model groups. The vascular cross?sectional area, plaque cross?sectional area and stenosis rate of the plaques in the two groups all differed significantly from those in the blank control group (P<0.05), but these parameters were similar between MPQ group and the model group (P>0.05). Compared with those in the blank control group, the hemodynamic parameters showed significant changes in MPQ and the model groups (P<0.05) but remained similar between the latter two groups (P>0.05).

CONCLUSIONCAS plaques can cause changes in hemodynamic parameters. Short?term therapy with MPQ does not affect the stability of the plaques or cause adverse effects on hemodynamics in cynomolgus monkeys with mild CAS plaques.

OBJECTIVETo provide a feasible and cost-effective next-generation sequencing (NGS) method for accurate identification of viral pathogens in clinical specimens, because enormous limitations impede the clinical use of common NGS, such as high cost, complicated procedures, tremendous data analysis, and high background noise in clinical samples.

METHODSViruses from cell culture materials or clinical specimens were identified following an improved NGS procedure: reduction of background noise by sample preprocessing, viral enrichment by barcoded oligonucleotide (random hexamer or non-ribosomal hexanucleotide) primer-based amplification, fragmentation-free library construction and sequencing of one-tube mixtures, as well as rapid data analysis using an in-house pipeline.

RESULTSNGS data demonstrated that both barcoded primer sets were useful to simultaneously capture multiple viral pathogens in cell culture materials or clinical specimens and verified that hexanucleotide primers captured as many viral sequences as hexamers did. Moreover, direct testing of clinical specimens using this improved hexanucleotide primer-based NGS approach provided further detailed genotypes of enteroviruses causing hand, foot, and mouth disease (HFMD) and identified other potential viruses or differentiated misdiagnosis events.

CONCLUSIONThe improved barcoded oligonucleotide primer-based NGS approach is simplified, time saving, cost effective, and appropriate for direct identification of viral pathogens in clinical practice.

Clinical Laboratory Techniques ; DNA Barcoding, Taxonomic ; DNA Primers ; Enterovirus ; classification ; genetics ; isolation & purification ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Influenza B virus ; genetics ; isolation & purification ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; methods ; Sequence Analysis, RNA ; methods

In this study, an analytical method based on ultraviolet spectroscopy was established for the rapid determination of nine components including isophorone, 4-methylene-isophorone, curcumenone, curcumenol, curdione, curzerenone, furanodienone, curcumol and germacrone in the first extraction process of Xingnaojing injection. 166 distillate samples of Gardeniae Fructus and Radix Curcumae were collected in the first extraction process of Xingnaojing injection. The ultraviolet spectra of these samples were collected, and the contents of the nine components in these samples were determined by high performance liquid chromatography. Least squares support vector machine and radial basis function artificial neural network were used to establish the multivariate calibration models between the ultraviolet spectra and the contents of the nine components. The results showed that the established ultraviolet spectrum analysis method can determine the contents of the nine components in the distillates accurately, with root mean square error of prediction of 0.068, 0.147, 0.215, 0.319, 1.01, 1.27, 0.764, 0.147, 0.610 mg•L⁻¹, respectively. This proposed method is a rapid, simple and low-cost tool for the monitoring and endpoint determination of the extraction process of Xingnaojing injection to reduce quality defects and variations.

OBJECTIVETo investigate the prevalence, etiology and clinical characteristics of adrenal lesions detected by abdominal computed tomography (CT).

METHODSThis retrospective study was conducted in patients with adrenal lesions detected by abdominal CT examinations in Nanfang Hospital between July, 2014 and June, 2015. The clinical data of the patients were collected for analysis of the demographics, comorbidities, imaging characteristics, biochemical profiles, clinical diagnosis and intervention.

RESULTSA total of 939 patients with adrenal lesions were identified from 19 004 patients undergoing abdominal CT scan over the defined period. The mean age of the patients was 53.2 years and 560 of the patients were male. Among the total cases with adrenal lesions, the percentages of cases with adrenal masses tended to increase progressively with age. Endocrine studies were done in 270 of the total patients, which identified non-functioning masses in 38.9%, primary aldosteronism in 16.3%, Cushing's syndrome in 4.1%, subclinical Cushing's syndrome in 7.0%, and pheochromocytomas in 7.0% of the cases. Adrenal incidentalomas was detected in 191 patients, with a detection rate of 1.0% among the overall patients undergoing abdominal CT scans. Imaging study detected adenomas (70.3%), cortical carcinomas (2.4%), and metastases (0.5%). Of 191 patients with adrenal incidentalomas, only 76 (39.8%) underwent endocrine evaluation, including 34 with nonfunctioning adrenal masses, 17 with pheochromocytoma, 7 with primary aldosteronism, and 5 with subclinical Cushing's syndrome.

CONCLUSIONs The overall detection rates of adrenal lesions and adrenal incidentalomas by abdominal CT were 4.9% and 1.0%, respectively, in our cohort of patients undergoing the examination over the defined period. Although most of the lesions were benign and nonfunctioning, malignant and functional lesions were also detected. As many as 60% of the patients with adrenal incidentalomas did not have hormonal testing. Clinicians need to have greater awareness of adrenal incidentalomas and standard protocol for its management should be established.

To understand the medication characteristics of Xingxue@Shuxuening solutions in the real world, multi-center, large-sample-size registration design method is adopted in this study. Between October 2012 and October 2015, hospitalized patients in 27 medical institutions who used Shuxuening and diagnosed as cerebral infarction were observed, including their general information, diagnosis information and medication information. Totally 9 473 cases of cerebral infarction were included, with old people as the majority, and males were slightly more men than females; they were complicated with coronary heart disease, vertebrobasilar artery insufficiency, transient brain ischemia and other cerebrovascular diseases; the medication duration was mostly within 14 days; the dosage was mainly 8-20 mL; 0.9%NS, 5%GS were the main solvent; commonly used drug combination was Ozagrel sodium injection, Shuxuetong injection, Alprostadil injection and other medicines for inhibiting platelet aggregation and blood vessel dilatation, and Cytidine diphosphate choline injection, Edaravone injection, Oxiracetam injection and other brain protective agents. The above results indicate that the clinical medication characteristics of Shuxuening are basically consistent with the package insert, except for the solvent; combined medicine is dominated by antiplatelet drugs; for old patients, various anticoagulant drugs shall selected according to their specific conditions to avoid the occurrence of adverse reactions.

To understand the clinical characteristics and distribution of combined treatment of Chinese and western medicine in diabetes deaths, the hospitalization information of diabetes deaths from HIS system of 20 national 3A-grade general hospitals. Then the frequency statistics and association rules analysis were used to analyze the general information, complications, combined treatment, death time and other information of the patients died from diabetes. The results showed that most of the diabetes deaths were of middle aged and elderly people, more often in males than females. The complications with higher incidence included hypertension, pulmonary infection, coronary heart disease, cerebral infarction and renal inadequacy. In combined treatment rules, western medicines insulin, cefuroxime, furosemide, dopamine, nikethamide and sodium bicarbonate were used in combination at highest frequencies, followed by the combinations of traditional Chinese medicines panax notoginseng, radix bupleuri and western medicines, and the combinations between Chinese medicines had the lowest use frequency. Most of the diabetes deaths were of middle aged and elderly people, more often in males than females. They mainly died from 3 pm to 5 pm and from 5 pm to 7 pm. Therefore, the diabetes deaths often had complications of cardiovascular and cerebrovascular diseases, and early prevention shall be noted in clinics; the clinical treatment plan was basically in accordance with the guidelines for clinical treatment of diabetes; the drugs with promoting blood circulation to remove blood stasis and soothing liver-qi stagnation effects were the common Chinese medicines in treatment of diabetes.

INTRODUCTIONThis study aimed to compare medication adherence and treatment persistence of patients on warfarin versus rivaroxaban in Singapore. A secondary objective was to identify significant covariates influencing adherence.

MATERIALS AND METHODSA retrospective cohort study was conducted where data from September 2009 to October 2014 was retrieved from the hospital electronic databases. Prescription records of rivaroxaban patients with 3 months or more of continuous prescription were extracted and compared against those of patients on warfarin. Primary outcome of adherence was determined based on the medication possession ratio (MPR), while treatment persistence was determined by outpatient clinic appointment gaps.

RESULTSA total of 94 rivaroxaban and 137 warfarin users were analysed by complete case analysis. The MPR of warfarin patients was lower than rivaroxaban patients by 10% (95% CI, 6.4% to 13.6%; P <0.0001). Also, there were more warfarin patients who had gaps in treatment persistence compared to those prescribed rivaroxaban (8.0% vs 1.1%; P = 0.03). Significant factors affecting medication adherence were age and duration of anticoagulant use. For every 10-year increase in age, MPR increased by 1.7% (95% CI, 0.7% to 2.8%). Similarly, for every year increase in duration of use, MPR increased by 1.8% (95% CI, 0.6% to 3.0%). Race, gender, concomitant medication and type of residence were not found to be significant covariates in the multivariable analysis.

CONCLUSIONPatients on rivaroxaban are likely to be more adherent to their prescribed oral anticoagulant with increasing age and duration of treatment influencing adherence.

Adult ; Age Factors ; Anticoagulants ; therapeutic use ; Databases, Factual ; Factor Xa Inhibitors ; therapeutic use ; Female ; Humans ; Male ; Medication Adherence ; statistics & numerical data ; Middle Aged ; Pulmonary Embolism ; drug therapy ; Retrospective Studies ; Rivaroxaban ; therapeutic use ; Singapore ; Venous Thrombosis ; drug therapy ; Warfarin ; therapeutic use