Objective To explore the function of fluorescent probe JC-1 in detecting the changes of mitochondrial membrane potential(△?m)in early apoptotic cells.Methods After 2-ME was used to induce MUTZ-1 cell apoptosis,cells were dyed with fluorescent probe JC-1,and then the changes of △?m in the early stage of apoptotic cells were analyzed by flow cytometry or detected under fluorescent microscope. Results The control cells with high △?m are those forming JC-1 aggregates in the inner membrane of mitochondria,thus showing orange-red fluorescence.2-ME caused decrease of △?m in MUTZ-1 cells,in which JC-1 maintains monomeric form,thus showing only green fluorescence.The decreases of △?m were in a time-dependent manner,which were significantly higher than those in control group(P

The study was aimed to investigate the mechanism of proliferation inhibition and apoptosis of MDS-RAEB MUTZ-1 cells induced by 2-methoxyestradiol (2-ME), the cell proliferation was determined by MTT assay, apoptosis rate was determined with annexinV-FITC/PI double staining and cell cycle was analyzed by flow cytometry (FCM) after MUTZ-1 cells were treated with different concentrations of 2-ME; the changes of morphologic features of MUTZ-1 cells were observed with Wright-Giemsa's staining; lactate dehydrogenase was determined by Beckman Counter; and agarose gel electrophoresis was used to verify whether 2-ME can induce apoptosis of MUTZ-1 cells. The results showed that 2-ME inhibited the proliferation of MUTZ-1 cells in a dose-and time-dependent manner and caused a sustained arrest at G(2)/M phase in MUTZ-1 cells; the typical apoptotic morphological features appeared in MUTZ-1 cells; the production of lactate dehydrogenase was up-regulated and the marked DNA ladder pattern of internucleosomal fragmentation was observed. It is concluded that the mechanism of proliferation inhibition and apoptosis of MUTZ-1 cells induced by 2-ME is probably related with the G(2)/M cell cycle arrest; 2-ME may be a potentially adjunctive anticancer drug useful to treat myelodysplastic syndrome.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Myelodysplastic Syndromes ; pathology

OBJECTIVETo study the association between gender and lung function in obese asthmatic children before and after inhaling corticosteroids (ICS).

METHODSA total of 102 asthmatic boys (including 57 normal-weight and 45 obese boys) and 88 asthmatic girls (including 46 normal-weight and 42 obese girls) received ICS treatment for one year. Pulmonary functions were evaluated before and after the treatment for forced expiratory volume in one second (FEV1), forced vital capacity (FVC), maximal expiratory flow 25 (MEF25) and maximal expiratory flow 50 (MEF50).

RESULTSThe asthmatic children in the 4 subgroups showed similar pulmonary functions before the treatment. After inhalation of methacholine, FVC% and FEV1% were improved in both normal-weight boys and normal-weight girls; FVC% and FEV1% in obese boys were improved after the treatment, but the obese girls showed improvement only in FVC%.

CONCLUSIONInhalation of methacholine can improve large airway function in asthmatic children, and gender affects the improvement to a lesser extent in normal-weight children. In obese asthmatic children, methacholine inhalation better improves lung functions in boys than in girls.

OBJECTIVETo study the association between body mass index (BMI) and lung function of asthmatic children after inhaling corticosteroids (ICS).

METHODSOne hundred and fifty-seven children with asthma were classified into obese (46 cases), over-weight (50 cases) and normal-weight groups (61 cases) based on BMI. All of the children received ICS for one year. Pulmonary functions were evaluated before and after treatment. Large airway function includes forced expiratory volume in one second (FEV1%) and forced vital capacity (FVC%). Small airway function includes maximal expiratory flow 25 (MEF25%) and maximal expiratory flow 50 (MEF50%).

RESULTSThe bronchial provocation test before treatment showed that the decline rate of pulmonary function (FVC%, FEV1%, MEF25% and MEF50%) in the obese group was higher than the normal-weight group after methacholine inhalation. After salbutamol inhalation, the improvement rate of the large airway (FVC%) and small airway (MEF25% and MEF50%) functions in the obese group was lower than the normal-weight group, and the improvement rate of small airway (MEF25% and MEF50%) function in the over-weight group was lower than in the normal-weight group. After treatment with ICS for one year, large airway function (FVC% and FEV1%) in the normal-weight group was higher than pre-treatment, however only FVC% in the normal-weight and obese groups was higher than pre-treatment. There was no significant difference in small airway function before and after treatment in all three groups.

CONCLUSIONSObesity can increase the sensitivity to methacholine and restrain the sensitivity to tosalbutamol in children with asthma. ICS can improve the large airway function in asthmatic children with normal body weight, but has no effect on small airway function. Obesity can restrain the effect of ICS on asthmatic children.

Administration, Inhalation ; Adrenal Cortex Hormones ; administration & dosage ; Asthma ; drug therapy ; physiopathology ; Body Mass Index ; Child ; Female ; Forced Expiratory Volume ; Humans ; Male ; Vital Capacity

OBJECTIVETo study the effect of 2-methoxyestradiol(2-ME) on telomerase activity expression and apoptosis in human myelodysplastic syndrome cells line MUTZ-1.

METHODSMUTZ-1 cells were incubated with different concentrations of 2-ME, apoptosis rate and cell cycle were measured by flow cytometry (FCM). Telomerase activity in MUTZ-1 cells was examined by telomeric repeat amplification protocol-Enzyme linked immunosorbent assay (TRAP-ELISA).

RESULTSThe FCM analysis showed that cells in G0/G1 phase and S phase were decreased, while in G2/M phase increased after exposed to 1,2 and 4 micromol/L of 2-ME for 12 hours (P < 0.05). 1 and 2 micromol/L of 2-ME had no notable effect on MUTZ-1 cells as compared with the control group (P > 0. 05). Cells incubated with 1, 2 and 4 micromol/L of 2-ME for 36 hours were induced apoptosis, the percentage of apoptosis was between (12.87 +/- 0.86)% and (21.82 +/- 1.71)% with a dose- and time- dependent manner. Telomerase activity was significantly inhibited in these concentration and negatively correlated with cell number in G2/M phase (r = -0.979, P = 0.021) and increased apoptosis (r = -0.970, P = 0.030 ), respectively. Moreover, the inhibition effect of telomerase activity was enhanced in a dose- and time- dependent manner.

CONCLUSIONS2-ME-induced apoptosis and inhibition of telomerase activity provide a possible mechanism for explaining the 2-ME's anticancer activity.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; Myelodysplastic Syndromes ; enzymology ; pathology ; Telomerase ; metabolism

To study the effects of sodium valproate (VPA) on human myelodysplastic syndrome cell line MUTZ-1. The cell proliferation was determined by MTT assay, apoptotic morphological features were observed by light microscopy and transmission electronmicroscopy, cell apoptosis and cell cycle shift were analyzed by flow cytometry (FCM). The results showed that VPA could inhibit the growth of MUTZ-1 cells in dose-and time-dependent manners. The typical apoptotic morphological features appeared in MUTZ-1 cells treated with 4 mmol/L VPA for 72 hours. Pyknosis of cells and nuclei, disintegration of nuclear chromatin and apoptotic body could be observed by light microscopy. Aggregation and margination of nuclear chromatin, concentration of plasm, increment of density and chromatin mass of irregular size could be observed by transmission electronmicroscope. The flow cytometric analysis indicated that the VPA could induce cell apoptosis, apoptosis rate increased in dose-dependent manner, ratio of cells at G(0)/G(1) phase increased and ratio of cells at S phase decreased in dose-dependent manner, the cells were arrested at G(0)/G(1) phase. It is concluded that the VPA can induce apotosis and inhibite proliferation of MUTZ-1 cells via arresting cells at G(0)/G(1) phase.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Dose-Response Relationship, Drug ; Humans ; Myelodysplastic Syndromes ; pathology ; Valproic Acid ; pharmacology

This study was aimed to investigate the expression of c-FLIPL, c-FLIPS and DLK1 mRNA in the patients with myelodysplastic syndrome (MDS) and its clinical significance. The mRNA expression of c-FLIPL, c-FLIPS and DLK1 in bone marrow mononuclear cells (BMMNC) of 16 patients with MDS and 3 controls were detected by RT-PCR. The results indicated that the expression of DLK1 mRNA was up-regulated in MDS, including RA and RAEB, as compared with controls (p < 0.05). There was no significant difference in expression of DLK1 between RA and RAEB patients (p > 0.05); the expression of c-FLIPL mRNA both in RA and RAEB patients was higher than that in controls (p < 0.05). There was no significant difference in expression of c-FLIPL between RA and RAEB patients (p > 0.05); the expression of c-FLIPS mRNA was not significantly different between MDS patients and controls (p > 0.05), but its expression in RAEB patients was significantly higher as compared with RA patients and controls (p < 0.05). It is concluded that the mRNA expressions of DLK1, c-FLIPL and c-FLIPS in MDS patients are abnormal, some of which may be useful as an important indicator for the evaluation of development in MDS.
Aged ; Bone Marrow Cells ; metabolism ; CASP8 and FADD-Like Apoptosis Regulating Protein ; genetics ; metabolism ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Myelodysplastic Syndromes ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo study the newborn's Apgar score in 'one minute' and relevant factors.

METHODSOne year inpatient woman from a Maternal and Child Health Hospital of Anhui province were selected by cluster sampling method and newborn asphyxia situation was investigated using Apgar score and self-designed questionnaire.

RESULTSThe Apgar score in 'one minute' which marking 8 to 10, 4 to 7 and 0 to 3 were found in 1875 (73.78%), 426 (16.77%) and 240 infants (9.45%) respectively. The average Apgar score in 'one minute' and five minutes were (7.69 +/- 2.27) and (9.01 +/- 1.89) respectively, The Apgar score in 'one minute' was significantly correlated with that of five minutes (Pearson coefficient correlation r = 0.841, P = 0.00). Ordinal regression analysis revealed that parturient age (OR = 1.04), being farmer (OR = 2.22), parity (OR = 1.26), assistant vaginal delivery (OR = 4.93), caesarean section (OR = 1.95), pregnancy-induced hypertension syndrome (OR = 1.42), albuminuria in gestational period (OR = 1.44), newborn being male (OR = 1.23), low birth weight (OR = 2.94), inborn abnormality (OR = 12.12), premature birth (OR = 1.22) and complications of delivery (OR = 5.04) were risk factors while the number of years under study (OR = 0.91), prenatal check-up (OR = 0.48), body length of newborn infant (OR = 0.88) and single birth (OR = 0.57) were protective factors.

CONCLUSIONApgar score in 'one minute' of newborn infant was affected by several factors as stated above. Health care program in earlier period toward community parturient should be strengthened in order to discover and control high risk factors of duration of pregnancy in earlier period.

Apgar Score ; Asphyxia Neonatorum ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Risk Factors ; Time Factors

The purpose of this study was to explore the mechanism underlying the regulation of 2-methoxyestradiol (2-ME)-induced cell apoptosis by mcl-1 and bax gene in myelodysplastic syndrome (MDS). The MUTZ-1 cells were pretreated with 2-ME; then the activity of caspases-3 was determined by fluorescent colorimetry; the mRNA expressions of apoptosis-related genes (mcl-1) and bcl-2-related X protein (bax) were determined by RT-PCR. The results showed that as compared with control, the 2-ME enhanced the activity of caspase-3 in MUTZ-1 cells in a dose-and time-dependent manners (p<0.05); along with increasing of 2-ME concentration, the expression of intracellular mcl-1 mRNA reduced (p<0.05), meanwhile the expression level of mcl-1 mRNA negatively correlated to the activity of caspase-3 at the corresponding time points (r=-0.992, p<0.01), but the expression of bax mRNA did not show significant change (p>0.05). It is concluded that 2-ME can regulate the apoptosis of MDS cells through the pathway of down-regulating the expression of mcl-1 mRNA and activating the caspase-3.
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Estradiol ; adverse effects ; analogs & derivatives ; Humans ; Myelodysplastic Syndromes ; metabolism ; pathology ; Myeloid Cell Leukemia Sequence 1 Protein ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

AIM To explore the effect of Heidihuang Pills on renal fibrosis in a rat model of chronic renal failure(CRF)and its mechanism.METHODS Wistar rats were randomly divided into the blank group for normal feeding and the model group for the establishment of CRF rat models by 5/6 nephrectomy.Subsequently,the successfully established rat models were randomly divided into the model group,the Heidihuang Pills group(10.43 g/kg),and the Heidihuang Pills+IGF-1R blocker(JB1)group for a regimen of 7-day subcutaneous injection of 18 μg/kg JB1 followed by gavage of 10.43 g/kg Heidihuang Pills.Eight weeks after the administration,the rats had their serum levels of Scr and BUN detected;their pathological changes of renal tissue observed by HE and Masson staining;their renal protein expressions of TGF-β,HIF-1α and α-SMA detected by immunohistochemistry;their renal protein expressions of IGF-1R and TGF-β detected by Western blot;and their renal mRNA expressions of IGF-1R and TGF-β detected by RT-qPCR.RESULTS Compared with the blank group,the model group displayed increased serum levels of Scr and BUN(P＜0.05);increased,degree of renal fibrosis,and renal fibrosis area(P＜0.05);increased renal expressions of TGF-β,HIF-1α,α-SMA proteins and TGF-β mRNA(P＜0.05);and decreased expressions of IGF-1R mRNA and protein(P＜0.05).Compared with the model group,the Heidihuang Pills group displayed decreased serum Scr and BUN levels(P＜0.05);decreased inflammatory cells in renal interstitium and the fibrosis degree(P＜0.05);decreased renal expressions of TGF-β,HIF-1α,α-SMA proteins and TGF-β mRNA(P＜0.05);and increased expressions of IGF-1R mRNA and protein(P＜0.05).However,the administration of JB1 could weaken the improvement effect of Heidihuang Pills on renal fibrosis in CRF rats(P＜0.05).CONCLUSION Heidihuang Pills can inhibit the renal fibrosis in CRF rats,and the inhibition process is related to up-regulated IGF-1 expression and promoted combination of IGF-1 and IGF-1R.