Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.

Objective:To investigate the effectiveness of heart rate deceleration capacity (DC) measurement in predicting the car-diotoxicity of malignant tumor patients treated with epirubicin-based chemotherapy. Methods:The clinical medical records, including CK-MB and cTnI levels and dynamic electrocardiogram (ECG) parameters before and after each chemotherapy cycle, of 140 patients treated with epirubicin-based chemotherapy were analyzed. Patients were divided into the DC>4.5 ms group and the DC≤4.5 ms group based on the calculated DC values. The CK-MB and cTnI levels and the dynamic ECG parameters of the two groups were compared af-ter two and four cycles of chemotherapy. Results:Patients in the two groups exhibited no statistically significant difference in their rele-vant clinical and pathological data before receiving chemotherapy (P>0.05). However, after four cycles of chemotherapy, the DC≤4.5 ms group showed a significantly greater increase in serum CK-MB and cTnI concentrations over the pre-chemotherapy levels compared with the DC>4.5 ms group. After two and four cycles of chemotherapy, the DC≤4.5 ms group also exhibited a significantly greater in-crease in mean heart rate (beats/min) and supraventricular and ventricular arrhythmia counts (times/24 h) over the pre-chemotherapy values compared with the DC>4.5 ms group (P<0.05). After four cycles of chemotherapy, 23 cases showed abnormally elevated cTnI levels in the DC≤4.5 ms group. In this group, patients with elevated cTnI level exhibited no statistically significant difference in CK-MB and cTnI concentrations, mean heart rates, and supraventricular and ventricular arrhythmia counts compared with those with nor-mal cTnI level before chemotherapy (P>0.05). However, the DC values of patients with elevated cTnI were significantly lower than those with normal cTnI level (P<0.05). Conclusion:The risk of epirubicin-induced cardiotoxicity increased with decrease in DC value. The DC test was shown to be an effective predictor of the risk of epirubicin-induced cardiotoxicity.

[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

OBJECTIVETo compare MR imaging with radiographs in detection of the erosions of the wrists in patients with rheumatoid arthritis (RA).

METHODSIn 50 wrists of 25 patients with RA, radiographs of AP projection and MR imaging examination were performed within 1 week. The MRI sequences were SE T1-weighted image, FSE T2-weighted image, and GE T2* weighted image in coronal plane. Gd-DTPA enhanced SE T1-weighted image with same parameters of pre-contrast administration SE T1WI was performed in 25 wrists, which showed prominent lesions in each 25 patients.

RESULTSIn 50 wrists of 25 patients with RA, bone erosions were found by MR images compared to radiographs in 46 versus 14 wrists, respectively. There were 170 sites of carpal erosions found by MR images, while radiographs found only 23 sites of carpal erosions. Among them triquetrum and lunate were the most frequently involved sites. Significant difference (P < 0.001) was statistically found between conventional radiography and MR imaging in detection of erosions of the wrists of RA. MR imaging also permitted demonstrating synovitis and joint effusion in the wrists. Gd-DTPA-enhanced images allowed distinguishing pannus changes from joint effusion.

CONCLUSIONMRI is superior to conventional radiography with respect to detection of bone erosion in wrists. For those whose conventional radiographs show suspected RA, MRI examination should be advised for the early diagnosis.

Adult ; Aged ; Arthritis, Rheumatoid ; diagnosis ; diagnostic imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Wrist Joint ; pathology

OBJECTIVEIn aid of MR imaging diagnosis of ankylosing spondylitis (AS), MRI findings of sacroiliac (SI) joints in volunteers were compared with those in patients with AS, with special reference to the fat findings.

METHODS18 normal volunteers and 52 AS patients underwent the MRI examinations with the same protocols, i.e., SE T1WI, FSE T2WI, GE T2*WI sequences were taken using a 0.3T imager. All the images were observed and analyzed by two radiologists.

RESULTSSymmetric or patchy distribution fat component either in sacrum and/or ilium was detected in 52.8% of SI joints of volunteers. Thin zone distribution of fat was seen in 35.6% of SI joints of AS patients, which was adjacent to the erosion and sclerosis. In another 25.0% of SI joints of AS patients, the distribution of fat component was found similar to that of the normal volunteers. Cartilage changes were detected in 85.6% of the SI joints of AS patients. In addition, bone erosion, sclerosis and subchondral bone marrow edema were also detected.

CONCLUSIONSubchondral fat infiltration associated with erosion and sclerosis should be considered as sacroiliitis. The knowledge of the characteristics of fat component could be helpful for evaluating sacroiliac joints.

Adipose Tissue ; pathology ; Adolescent ; Adult ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Sacroiliac Joint ; pathology ; Spondylitis, Ankylosing ; diagnosis ; pathology

Objective To evaluate the correlation of maternal FTO expression and insulin secretion levels in pregnant women.Methods In outpatient of Zhuhai Municipal Maternal and Child Healthcare Hospital,60 cases of pregnant women with gestational diabetes mellitus (GDM) and 60 cases of healthy pregnant women were recruited in this study from March 2016 to August 2016.Each pregnant woman was taken two tubes of venous blood.Lymphocytes was isolated from one tube of anticoagulated whole blood.The expression of FTO mRNA in lymphocytes was investigated using RT-PCR.Serum was isolated from another tube of non-anticoagulant whole blood.Its FTO protein was investigated using ELISA,insulin was investigated using chemiluminescence method,and GLU,CHO,TG,HDL-C and LDL-C were investigated by automatic biochemical analyzer.The correlation of the biochemical indicators were analyzed.Results The expression level of FTO mRNA,FTO protein,fasting glucose data,OGTT at 1 h and 2h glucose levels in the case group were 0.17 ％ ± 0.10 ％,61.32 ± 25.23 pg/ml,4.71 ±0.54 mmol/L,10.70±1.36 mmol/L and 8.97 ± 1.60 mmol/L respectively.The values of these biochemical indicators in control group were 0.11％±0.07％,51.47±22.97 pg/ml,4.41± 0.28 mmol/L,8.05±1.04 mmol/L and 6.56±0.75 mmol/L respectively.The differences were statistically significant (t =3.876,2.236,3.817,11.964 and 10.578,P＜0.05).The insulin levels were 6.83±9.76 mU/L in the case group,and 13.15±13.99 mU/L in control group.The differ ences was statistically significant too(t=-2.869,P＜0.05).FTO expression level was positively correlated with FTO protein,OGTT 1h glucose and OGTT 2h glucose levels (r=0.232,0.292,0.242,all P＜0.05),and it was negatively correlated with insulin levels(r=-0.185,P＜0.05).There was no correlation between FTO and fasting glucose expression levels (P ＞0.05).Conclusion The expression of FTO level in pregnant women with GDM was higher than that in healthy pregnant women.The relative expression of FTO mRNA was negatively correlated with insulin.And the regulation of glucose metabolism might be effected by insulin.

Objective To investigate the potential of adult mesenchymal stem cells (MSCs) derived from human bone marrow to undergo cardiomyogenic differentiation after exposure to 5-azacytidine (5-aza) in vitro. Methods A small bone marrow aspirate was taken from the iliac crest of human volunteers, and hMSCs were isolated by 1.073g/mL Percoll and propagated in the right cell culturing medium as previously described. The phenotypes of hMSCs were characterized with the use of flow cytometry. The hMSCs were cultured in cell culture medium (as control) and medium mixed with 5-aza for cellular differentiation. We examined by immunohistochemistry at 21 days the inducement of desmin, cardiac-specific cardiac troponin I (cTnI), GATA 4 and connexin-43 respectively. Results The hMSCs are fibroblast-like morphology and express CD44+ CD29+ CD90+ / CD34- CD45- CD31- CD11a. After 5-aza treatment, 20-30% hMSCs connected with adjoining cells and coalesced into myotube structures after 14days. Twenty-one days after 5-aza treatment, immunofluorescence showed that some cells expressed desmin,GATA4, cTnI and connexin-43 in 5,10 μmol/L 5-aza groups, but no cardiac specific protein was found in neither 3μmol/L 5-aza group nor in the control group. The ratio of cTnI positively stained cells in 10 μmol/L group was higher than that in 5 μmol/L group (65.3 ± 4.7% vs 48.2 ± 5.4%, P ＜ 0.05). Electron microscopy revealed that myofilaments were formed. The induced cells expressed cardiac-myosin heavy chain (MyHC) gene by reverse transcription-polymerase chain reaction (RT-PCR). Conclusions Theses findings suggest that hMSCs from adult bone marrow can be differentiated into cardiac-like muscle cells with 5-aza inducement in vitro and the differentiation is in line with the 5-aza concentration. (J Geriatr Cardiol 2004;1(2) :101-107. )

Serum cytokine concentrations were determined in patients with Graves'disease(GD)before and after treatment of thiamazole.Serum soluble interleukin-2 receptor(sIL-2R)level was markedly raised before treatment as compared with normal subjects and returned to normal after 3 month therapy.Serum IL-8 level was lowered in GD patients and showed no change after 3 month therapy.No relationship between IL-8 and other markers was found.Serum IL-1?,IL-6 and TNF-?levels in GD patients showed no difference with those in normal controls.

Objective To explore the doctor-nurse integrated health care management model in perioperative nursing management of transvaginal natural orifice transluminal endoscopic surgery (NOTES). Methods A total of 70 female patients underwent transvaginal NOTES. All patients were randomly divided into the control and the study groups. The control group adopted the traditional model of care, and the study group implemented a layered panel system of integrated health care nursing mode, which consisted of three lines (leader), second line, front-line doctors, anesthesiologists consisting of the doctor series and secondary responsibility leader, a responsibility of nurses, operating room nurses laparoscopy group traveling together constituted a series of nurses′ health care integration of the health care team. The average length of stay, preoperative waiting days, awareness of the disease, early postoperative activities and reasonable diet, patient satisfaction with care, doctors′ satisfaction with nurses′ operation and overall satisfaction with health care work were compared. Results The preoperative waiting days, average length of stay, surgical satisfaction, patients′ satisfaction, doctors-nurses′overall satisfaction in the study group were (3.95±0.51) d, (11.58±1.83) d, (95.48±3.82)%, (99.26± 0.61)%, (96.33±0.29)%, the control group were (5.50±0.68) d, (15.79±2.14) d, (82.08±7.21)%, (92.18± 3.54)%, (80.03 ± 5.88)%, the differences were statistically significant (t=-4.431-6.172, P < 0.05). The patients awareness of the disease, early postoperative activities and reasonable diet (postoperative eating time) in the study group were (92.77 ± 0.59)%, (1.60 ± 0.63) d, (2.20 ± 0.54) d, the control group were (93.15 ± 0.47)%, (1.90 ± 0.42) d, (2.40 ± 0.33) d, the difference was not statistically significant (t=-0.898,-0.771,-0.201, P>0.05). Conclusions The implementation ofdoctor-nurse integrated health carenew model can optimize the collocation of medical health care integration, enhance the quality of care, improve patient experience, and improve patients′and doctors′satisfaction for nursing work.