OBJECTIVE: To explore the effect of melatonin(Mel) on the proliferation, apoptosis and expression of bcl-2 in oxidized low-density lipoprotein(ox-LDL)-induced endothelial progenitor cells (EPC) from human umbilical cord blood in vitro. METHODS: Total mononuclear cells were isolated from human umbilical cord blood in vitro by Ficoll density gradient centrifugation, and the cells were plated on fibronectin-coated culture dishes. After 7 days, the attached cells were divided into 7 groups: a control group (normal cells), 3 ox-LDL groups[the attached cells were incubated with different concentrations of ox-LDL(5,10,and 20mg/L) for 24 hours], and 3 Mel groups[the attached cells were incubated with different concentrations of Mel (0.5,1.0, and 2.0 mmol/L) respectively for 24 hours before incubation with 10 mg/L ox-LDL]. EPC was identified by examining the expression of CD34, vascular endothelial growth factor receptor-2(VEGFR-2) and CD133 under a laser scanning confocal microscope. We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the effect of Mel and ox-LDL on the multiplication ability of EPC. Flow cytometry was used to detect the apoptosis. The expressions of Bcl-2 mRNA and protein were detected respectively by RT-PCR and immunohistochemistry technology. RESULTS: After being exposed to the ox-LDL, the proliferation of EPC in the 3 ox-LDL groups was lower, and the apoptosis rate was higher than that in the control group in a dose-dependent manner (P<0.01); Mel was added at different concentrations before the ox-LDL incubation, and the cells in the 3 Mel groups showed higher proliferation and lower apoptosis rate than those of the 3 ox-LDL groups (P<0.01). Expression of Bcl-2 mRNA and protein of EPC in the 3 Mel groups was higher than that in the 3 ox-LDL groups (P<0.01). CONCLUSION Ox-LDL can inhibit the proliferation of EPC and promote the apoptosis of the cells by down-regulating the bcl-2 expression. Mel can inhibit these effects of ox-LDL.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; Humans ; Lipoproteins, LDL ; adverse effects ; Melatonin ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stem Cells ; cytology ; drug effects

OBJECTIVE: To investigate the culture of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases (CHD) before and after percutaneous coronary intervention (PCI), and to observe the cells shape and determine the cell number and proliferation activity. METHODS: Ninety-five patients were divided into a CHD group(n=65) and a control group (n=30). The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF.EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colony-forming units(CFU) was counted by phase-contrast microscope. RESULTS: The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. How-ever, the number of CFUs did not change in patients before and after the PCI. CONCLUSION PCI can increase endothelial progenitor cells in patients after the PCI; but 4 days after the PCI, this increase will not exist.
Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Cell Adhesion ; Cell Count ; Cell Movement ; Cells, Cultured ; Coronary Disease ; blood ; therapy ; Endothelial Cells ; pathology ; Female ; Humans ; Male ; Middle Aged ; Stem Cells ; pathology

Microvesicles transport special proteins, micro RNA and DNA segments, which provides new access to intercellular communication. Tumor-derived membrane microvesicles (TMV) are involved in the tumor progress by transporting tumor-derived proteins, delivering microRNA to surrounding normal cells to alter their phenotype and promoting reverse transcription to interfere gene stability and to create tumor microenvironment. TMV also play crucial roles in tumor angiogenesis and matrix degradation, which facilitates malignant cell metastasis. TMVs are also involved in escaping immunological surveillance by intensifying the function of suppressor T cell and inducing apoptosis of cytotoxic T cells. On the other hand, microvesicles carry tumor antigens and can be used for development of tumor vaccines; some new vaccines such as AEX and DEX are under early clinical trials. Circulating microRNA and DNA segments in body fluid can be a new potential biomarker for cancer diagnosis and prognosis. Purification of microvesicles needs to be further improved, which is important for identification of microvesicles and their subtypes.
Cell Communication ; Cytoplasmic Vesicles ; chemistry ; physiology ; ultrastructure ; Humans ; Neoplasms ; pathology ; physiopathology ; Neoplastic Processes ; Neovascularization, Pathologic ; Tumor Microenvironment

OBJECTIVETo investigate the relationship between expression of vascular endothelial growth factor (VEGF) and pathogenesis of acute leukemia (AL).

METHODSVEGF mRNA expression was analysed by using semi-quantitative RT-PCR technique. VEGF level of culture supernatant of the bone marrow mononuclear cell (BMMNC) was detected by ELISA. The influence of culture supernatant of the BMMNC on proliferation of human umbilical vein endothelial cell line huECV304 in vitro was determined by MTT assay.

RESULTSThe median VEGF/beta-actin expression level of BMMNC in newly diagnosed untreated AL group was higher (0.86) than that in remission group (0.41) and normal control group (0.39) (P < 0.01, both), but was no difference from the relapse group (1.02, P > 0.05). Among the patients with newly diagnosed untreated AL, VEGF/beta-actin level in AML (1.03) was statistically higher than that in ALL (0.61) (P < 0.05). The VEGF level in 72 h culture supernatant of BMMNC was higher in newly diagnosed untreated AL group (91.48 ng/L) than in remission group (31.91 ng/L) and normal control group (28.71 ng/L) (P < 0.01). In ALL group (32.76 ng/L), the VEGF level of 72 h culture supernatant was much lower than that in AML group (173.49 ng/L) (P < 0.01). The proliferation of huECV304 cells after co-cultured with the 72 h culture supernatant of AML BMMNC with addition of anti-VEGF antibody was inhibited notably compared with culture supernatant alone (P < 0.01). However, no such result was found in ALL group (P > 0.05).

CONCLUSIONThe mRNA expression and secretion of VEGF in AML cells were higher than those in ALL cells. The regulators that participated angiogenesis were different in ALL and AML patients.

Acute Disease ; Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Female ; Humans ; Leukemia ; metabolism ; Male ; Monocytes ; metabolism ; RNA, Messenger ; genetics ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; pharmacology

OBJECTIVETo study changes of left ventricular remodeling (LVR) in hypertension patients with carotid atherosclerosis (CAS) of phlegm-dampness syndrome (PDS).

METHODSDoppler ultrasonography data of CAS were observed in 223 hypertension patients with CAS (as the hypertension group, including 119 patients of the PDS group and 104 of the non-PDS group), 81 CAS patients with non-hypertension, and 19 non-hypertension non-CAS patients (as the control group). The difference in the degree of LVR was compared among the above groups.

RESULTSThe left ventricular posterior wall thickness (LVPWT), inter ventricular septum thickness (IVS), E/A were higher in the hypertension group than in the non-hypertension group (P < 0.05). The left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic diameter (LVESD), stroke volume (SV) were higher in the soft plaque hypertension group and the soft plaque non-hypertension group than in the hard plaque group, the thickening intimal group, and the normal intimal group (P < 0.01 , P < 0.05). The LVEDD, LVESD, and SV were higher, and the ejection fraction (EF) was lower in the PDS hypertension group than in the non-PDS hypertension group (all P < 0.05). Of them, LVEDD, LVESD, and SV were higher in the soft plaque group than in the hard plaque group (P < 0.01), the thickening intimal group (P < 0.01) and the normal intimal group (P < 0.05). There was no statistical difference in PDS hypertension between the soft plaque group and the hard plaque group (P > 0.05).

CONCLUSIONThe hypertension patients with CAS of PDS might be correlated to LVR, and LVR was more obviously in the soft plaque patients.

Aged ; Aged, 80 and over ; Carotid Artery Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Case-Control Studies ; Female ; Humans ; Hypertension ; diagnosis ; diagnostic imaging ; physiopathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ultrasonography ; Ventricular Remodeling

Objective To determine the expression of membrane-bound B lymphocyte stimulator (BLyS) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from individuals with systemic lupus erythematosus (SLE),and to investigate the role of interferon-?(IFN-?) on the expression of BLyS.Methods PBMCs were obtained from 25 SLE patients (mean age of 31+14) and 20 healthy volunteers (mean age of 28?10).They were randomized into IFN-?(5 ng/ml) group and control group.PBMCs were col- lected at 0,6,12 and 24 h for BLyS mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR).PBMCs were also collected at 72 h for membrane-bound BLyS protein detection using flow cy- tometry (FACS) and direct immunofluorescence.Results①The expression of BLyS mRNA and membrane- bound protein in PBMCs was significantly higher in individuals with SLE compared with healthy controls (P＜0.05);②IFN-?enhanced BLyS mRNA expression in PBMCs in both healthy controls and SLE patients,with the greatest effect at 6 h (stimulated vs unstimulated,0.42?0.19 vs 0.25?0.14,P＜0.01;0.59?0.28 vs 0.44?0.21,P＜0.01 );③IFN-?also increased the expression of membrane-bound BLyS protein in both healthy con- trols and individuals with SLE (FACs,mean fluorescence intensity,4.5+3.0 vs 3.7~2.6,P

From original concept and literature of acupoint, the concept and clinical significance of ashi method is discussed, which clarifies that the essence of ashi method is to locate the acupoints by patients' sensitivity on force. The clinical application of heat-sensitive moxibustion has illustrated that positioning method of this therapy is based on the appearance of heat-sensitive moxibustion sensation. Although both types are based on patients' feeling, positioning method of heat-sensitive moxibustion stands on a new angle and uses a new method to locate acupoint. Therefore, it is believed that the positioning method of heat-sensitive moxibustion is the inheritance and development of ashi method.
Acupuncture Points ; China ; History, Ancient ; Humans ; Medicine in Literature ; Moxibustion ; history ; methods ; Sensation

OBJECTIVETo study effects of different interval year on Panax notoginseng plant soils middle and micro element content.

METHODThe dynamic change of Ca, Mg, Mn, Cu, Zn, B from Mabai, Matang, Gumu and Panlong were determined under different planting patterns (new soil, interval 5 years soil and continuous cropping soil).

RESULTAll the micro elements (except Ca, Mg) of interval 5 years soil and continuous cropping soil were significantly higher than those of new soil. All the middle and micro elements (except B) of interval 5 years soil were significantly higher than those of the continuous cropping soil. Planting patterns had remarkable influence on the content of Mn, Cu, B, but not Zn Ca, Mg. Cu, Ca under the 3 planting patterns, and Zn under the continuous cropping pattern did not show significant quarter changes. B content increased with the elongation of implantation time. Zn in new soil and interval 5 years also increased with prolonging of planted time. Mg, Mn and Cu content reached to peak value on April next year, and reached to minimum on the end of this experiment. Compared with new soil, the proportion of Mn, Cu in total elements increased by 29%, 114%, Mg, B decreased by 18%, 38%, Zn and Ca changed slightly of interval 5 years soils; In continuous cropping soil, Mn, Cu and B increased by 50%, 120%, 22%, respectively, but Zn, Ca, Mg had no significant change.

CONCLUSIONContinuous cropping pattern could not induce the deficient of soil middle and micro elements, and thereafter might not result in continuous cropping obstacles. But the imbalance proportional of soil middle and micro elements in P. notoginseng plant soils may be one of the main reasons for continuous cropping obstacles.

China ; Kinetics ; Panax notoginseng ; growth & development ; Soil ; chemistry ; Time Factors ; Trace Elements ; chemistry

OBJECTIVETo study effects of different interval year on Panax notoginseng plant soils macro element content.

METHODThe dynamic change of total N, P, K and available N, P, K in soil from Mabai, Matang, Gumu and Panlong was determined under different planting patterns (new soil, interval 5 years soil and continuous cropping soil).

RESULTContents order of soil total N, P and available N, P were interval 5 years soil > continuous cropping soil > new soil. No significant quarter change on soil total N was found, but the other three showed inverted "v" curve, and the peak value appeared on April 2010. Content of soil total K did not change significantly, but the available K content order was new soil > continuous cropping soil > interval 5 years soil, the quarter change was similar as soil available N or P. The soil total N, P, K and available N, P, K were different of the 4 monitoring sites under the 3 interval planting modes. There was a significant correlation between soil total P and available P under all these 3 interval planting modes, but N and K. The propitiation of N-P-K of new soil, interval 5 years soil and continuous cropping soil were 1: 0.4: 2.4, 1: 0.4:1.4, 1:0. 4:2.0, respectively.

CONCLUSIONContinuous cropping pattern induce the accumulation of P, but deficient of K. The imbalance proportion of N, P and K was one of the incentives of continuous cropping induced obstacles. Strengthen the research of optimum proportion of soil N, P and K, and then eliminate continuous cropping obstacles by means of formulated fertilization is the future research direction.

Agriculture ; methods ; Breeding ; China ; Drugs, Chinese Herbal ; analysis ; Fertilizers ; analysis ; Nitrogen ; analysis ; metabolism ; Panax notoginseng ; chemistry ; growth & development ; metabolism ; Phosphorus ; analysis ; metabolism ; Potassium ; analysis ; metabolism ; Quality Control ; Soil ; chemistry

OBJECTIVE: To analyse the clinical and laboratory characteristics of antinuclear antibody (ANA) positive rheumatoid arthritis (RA) patients. METHODS: The clinical and laboratory data of 428 RA cases from Department of of Rheumatology and Immunology Peking University Third Hospital from Jan 2013 to Dec 2018 were collected and used to analyse characters between ANA positive group and ANA negative group. T test was used for the quantitative data in accordance with normal distribution. Wilcoxon rank sum test was used for the quantitative data of non normal distribution. The qualitative data were analyzed by chi square test. But while 1≤theoretical frequency < 5, chi square test of corrected four grid table was used. And Fisher exact probability method was used when theoretical frequency < 1. RESULTS: The number of ANA positive group was 231 (54%). The female rate was obviously higher in ANA positive group (82.7% vs. 63.5%, χ²=20.355, P < 0.01). The rate of metatarsophalangeal joints (MTPJs) involvement was lower in ANA positive group (22.1%) than in ANA negative group (33.0) (χ²=6.414, P < 0.05). The incidence of secondary Sjögren's syndrome (sSS) was much higher in ANA positive group(19.5% vs. 4.1%, χ²=23.300, P < 0.01). The positivity of rheumatoid factor (RF), as well as the positivity of anti-cyclic citrullinated peptide(CCP) antibody was much higher in ANA positive group (77.1% vs. 53.8%, χ²=25.743, P < 0.01, 74.9% vs. 59.4%, χ²=11.694, P < 0.01, respectively). The levels of immunoglobulin G (IgG) and immunoglobulin M (IgM) of ANA positive group were higher [(15.1±5.1) g/L vs. (13.8±5.3) g/L, t=2.359, P < 0.05, 1.25 (0.92) g/L vs. 1.05 (0.65) g/L, Z=-3.449, P < 0.01, respectively]. But the levels of hemoglobin (Hb) and platelet (PLT) was lower in ANA positive group[(109.64±17.98) vs. (114.47±18.48) g/L, t=-2.734, P < 0.01; (266.4×10⁹±104.6×10⁹) vs. (295.9×10⁹±100.1×10⁹) /L, t=-2.970, P < 0.01, respectively]. CONCLUSION The incidence of sSS was obviously higher in ANA positive group than in ANA negative group. Serum IgG of ANA positive group was higher, but Hb and PLT were lower.
Antibodies, Antinuclear ; Arthritis, Rheumatoid/epidemiology* ; Autoantibodies ; Female ; Humans ; Laboratories ; Peptides, Cyclic ; Rheumatoid Factor