1.Role of p38MAPK signaling pathways in the apoptosis of C2C12 myoblast cells subjected to cyclical stretch
Zhen TIAN ; Zhuli YANG ; Wenmin JIA ; Xiao YUAN ; Jing QIU ; Yu DA ; Yanxiao DU ; Jiangbo YU ; Yue ZHANG ; Wen LIU
Chinese Journal of Tissue Engineering Research 2011;15(15):2751-2754
BACKGROUND: Because of complicated physiological environment and difficulty to control experimental conditions, it is difficult to get satisfactory results from in vivo studies of cell mechanics.OBJECTIVE: To study the action and mechanism of p38MAPK signaling pathways on myoblast apoptosis based on successful construction of in vitro mechanical stimulation models.METHODS: The C2C12 cells cultured in vitro were divided into control group and SB203580 treatment group. Cyclic tensile stress was applied on the C2C12 myoblast cells for 0, 6, 12 and 24 hours in each group. The Flexcell Strain Unit-5000T was used to expose C2C12 myoblast cell to an equiaxial cyclic of 15% magnitude and a frequency of 10 cycles/min, each cycle including the 3 s stretch and 3 s relaxation. Hoechst 33258 fluorescent staining and optical microscope were used to detect cell apoptosis. RT-PCR, flow cytometric analysis were used to observe the apoptosis of C2C12 myoblast cells and Western blotting were used to detect the activity of p38MAPK and p-p38MAPK. RESULTS AND CONCLUSION: The optical microscope tested the change in the morphology. Hoechst 33258 staining showed that after treatment with cyclic stress, the cell took the typical appearance of apoptosis with chromatin condensation and apoptotic bodies. RT-PCR and flow cytometry showed that with the extension of time the rate of the apoptosis of C2C12 myoblast cell increased. And cells imposed SB203580 before imposing cyclical tensile stress, the results showed that the apoptosis was markedly affected, and the p-p38MAPK expression declined apparently. These findings demonstrate that p38MAPK signaling pathways in stress mediated into C2C12 myoblast cell apoptosis plays an important role.
2.Significance of Determination of Argyrophilic Proteins in Nucleolar Organizer Regions of Blood T Lymphocytes before and after Complete Remission in Children with Acute Leukemia
wei, LI ; ling-zhen, DU ; yao-li, WANG ; ding-zhong, QIU ; wen-quan, XIA ; yi-qun, HU
Journal of Applied Clinical Pediatrics 2006;0(15):-
Objective To detect the argyrophilic proteins in nucleolar organizer regions(Ag-NORs) that express rDNA and rRNA proliferation of T lymphocytes before chemotherapy and after complete remission(CR) in children with primary acute leukemia(AL).Methods The argyrophilic granules area of NOR/nuclear area(I.S%) of T lymphocytes was detected by image analysis system in peripheral blood of 42 patients before chemotherapy and after CR and 30 normal children.Results I.S% in the patients before chemotherapy(5.06%?1.36%) were significantly lower than those in the healthy donors(7.51%?1.06%)(t=8.238 P0.05).Conclusion These results suggest that decrease of Ag-NORs expresses the evidence for tumour induced suppression of immune function of T cells in children with AL prior to treatment.
3.Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells.
Zhen-hua LI ; Wen-hai ZHAO ; Qiu-li ZHOU
China Journal of Orthopaedics and Traumatology 2011;24(3):245-248
OBJECTIVETo study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.
METHODSFifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.
RESULTSDCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).
CONCLUSIONThe VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.
Animals ; Antlers ; chemistry ; Cartilage ; drug effects ; metabolism ; pathology ; Female ; Glutathione ; blood ; Male ; Nitric Oxide ; blood ; Osteoarthritis ; blood ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Peptides ; pharmacology ; Rabbits ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; blood
5.The expression and clinical implication of plasma miR-328 in patients with atrial fibrillation.
Xi WANG ; Chun-guang QIU ; Zhen-wen HUANG ; Zhan-ying HAN ; Wen-jie LU ; Xiao-jie CHEN
Chinese Journal of Cardiology 2013;41(2):126-129
OBJECTIVETo observe the expression and clinical implication of plasma miR-328 in patients with atrial fibrillation (AF).
METHODSFifty-eight patients with AF (AF group: 17 paroxysmal AF, 21 persistent AF, and 20 permanent AF) and 15 healthy volunteers (Control group) were included. General clinical data and related biochemical parameters were collected. Plasma miR-328 levels were detected with quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The correlation between plasma miR-328 and AF risk factors was analyzed.
RESULTS(1) Compared with the control group, the expression level of plasma miR-328 was significantly elevated in AF group (fold 7.72 ± 9.32) (P < 0.05). (2) In AF group, the expression of plasma miR-328 was significantly different in different type of AF[paroxysmal AF with (1.98 ± 0.81), persistent AF with (6.57 ± 5.82) and permanent AF with (13.47 ± 12.29)] (P < 0.05), and which was increased in proportion to the duration of AF. (3) There was a positive correlation between plasma miR-328 level and left atrial diameter in the AF group (r = 0.310, P < 0.05).
CONCLUSIONmiR-328 expression is significantly increased in patients with AF, which may be involved in the atrial remodeling process of AF.
Aged ; Atrial Fibrillation ; blood ; Case-Control Studies ; Female ; Humans ; Male ; MicroRNAs ; blood ; Middle Aged
6.Clinical observation on acupuncture for treatment of abdominal postoperative pain.
Yan-Hua XU ; Qiu-Yun WANG ; Zhen-Bao YU ; Yan-Wen HE
Chinese Acupuncture & Moxibustion 2010;30(11):904-906
OBJECTIVETo compare the therapeutic effects between acupuncture based on syndrome differentiation and analgesic on abdominal postoperative pain.
METHODSOne hundred cases of abdominal postoperative pain were randomly divided into two groups, 50 cases in each one. In acupuncture group, the treatment was applied according to meridian differentiation and point selection on the affected meridian. Ashi points near to the incision as the main points and those closely connected with Zangfu functions were selected, such as Yanglingquan (GB 34), Taichong (LR 3) and Zusanli (ST 36), etc. In medication group, muscular injection of Bucinnazine was administered. The severity of pain was evaluated with Visual Analogue Scale (VAS) before and after treatment.
RESULTSThe remarkably effective rate in acupuncture group was 60.0% (30/50), which was markedly better than that 28.0% (14/50) in medication group (P < 0.01). VAS scores in 30 min and 4 h after treatment as well as 24 h after operation in two groups were all reduced remarkably as compared with those before treatment (all P < 0.01), indicating the satisfactory analgesia in treatment. VAS scores in acupuncture group were lower apparently than those in medication group in 30 min and 4 h after treatment (P < 0.01,P < 0.05).
CONCLUSIONAcupuncture has quick analgesia in treatment of abdominal postoperative pain, which is superior to muscular injection of Bucinnazine because of its advantages of long-term and significant efficacy.
Abdomen ; surgery ; Acupuncture Analgesia ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pain Measurement ; Pain, Postoperative ; therapy ; Young Adult
7.Clinical analysis of upper airway resistance syndrome and obstructive sleep apnea-hypopnea syndrome in children.
Jian-wen ZHONG ; Da-bo LIU ; Zhen-yun HUANG ; Zong-yu TAN ; Shu-yao QIU ; Jie YU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2009;44(6):464-468
OBJECTIVETo analyze the similarities and differences of the clinical manifestations between the children with upper airway resistance syndrome (UARS) and obstructive sleep apnea-hypopnea syndrome (OSAHS), and to explore the clinical features and characteristics of sleep respiratory parameters.
METHODSUsing the double-blind method, all children were diagnosed as UARS or OSAHS through the polysomnography test and the results of all children were analyzed by a sleep technician and an otolaryngologist. Another ENT doctor recorded their clinical and physical examination in detail.
RESULTSPolysomnography showed that the apnea-hypopnea index (AHI) and the lowest oxygen in 253 children with OSAHS were 3.60[2.00;7.55] times/h and 0.90[0.85;0.91], and were 0.90[0.50;1.10] times/h and 0.95[0.92;0.96] in 102 children with UARS, the difference of the two groups by rank test was statistically significant. The proportion of UARS and OSAHS was more common during preschool period than during school-age period. The chief complaint in two groups was sleep snoring, and the main symptoms were sleep restless, attention deficit/hyperactivity and breath with mouth open. The incidence rate of above symptoms were as follows, 94.1%, 72.5%, 62.7% and 37.3% in children with UARS, 92.9%, 78.7%, 57.7% and 45.5% in children with OSAHS. The difference was not significant by chi-square test (P>0.05). Tonsil and adenoid hypertrophy were also observed in the two groups, the difference was not significant (chi2 = 0.27, P= 0.87). However, the children with OSAHS were more apt to have the sleep apnea than with UARS, the difference was statistically significant (chi2 = 34.07, P<0.001).
CONCLUSIONSThe clinical manifestations of two groups are similar, the difference between UARS and OSAHS can not be determined by the patient's clinical performance. Sleep apnea can be more easily observed in children with OSAHS than that in UARS, the final diagnosis is based on polysomnography.
Adolescent ; Airway Resistance ; Child ; Child, Preschool ; Double-Blind Method ; Female ; Humans ; Male ; Polysomnography ; Sleep Apnea, Obstructive ; epidemiology ; physiopathology
8.Association of carotid atherosclerosis and blood pressure variation.
Ao-Ya LIU ; Zhen-Qiu YU ; Wen-Hua WANG ; Xiao-Hui YANG ; Qiang YONG
Chinese Journal of Cardiology 2011;39(6):484-487
OBJECTIVETo investigate the relationship of carotid artery plaque and blood pressure variation and cardiovascular risk factors.
METHODSWe retrospectively analyzed clinical data of in-patients treated in the department of hypertension between April 2009 and June 2010. Information on carotid ultrasonography and other clinical date were obtained from 408 patients. All patients were monitored by ambulatory blood pressure.
RESULTS(1) Carotid artery determined in plaque was 55.3%, there was no differences between men and women. However, the carotid artery plague was associated positively with age. Increased age was associated with a significantly increased positive rate. (2) Cardiovascular risk factors and carotid artery plaque: carotid artery plaque was associated with duration of disease, fasting blood sugar, total cholesterol, and low density lipoprotein-cholesterol. (3) 24 h ambulatory blood pressure and carotid artery plaque: the prevalence of carotid artery plaque increased with increasing coefficient of systolic variation (P = 0.001). There was no correlation between the coefficient of diastolic variation and the prevalence (P = 0.644).(4) Multivariate regression analysis indicated that carotid artery plaque was associated with duration of hypertension, 24 h mean systolic blood pressure, and coefficient of variation of 24 h blood pressure (P < 0.05).
CONCLUSIONCarotid atherosclerosis is independently associated with coefficient of variation of blood pressure, especially with coefficient of variation of systolic blood pressure.
Adult ; Aged ; Blood Pressure ; Carotid Artery Diseases ; etiology ; Female ; Humans ; Hypertension ; complications ; pathology ; physiopathology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors
9.Diagnosis and treatment of obstructive sleep apnea hypopnea syndrome in children with risk factors.
Da-bo LIU ; Shu-yao QIU ; Jian-wen ZHONG ; Zhen-yun HUANG ; Qian CHEN ; Zong-yu TAN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2008;43(12):924-928
OBJECTIVETo analyse the clinical features of children with obstructive sleep apnea-hypopnea syndrome (OSAHS), accompanying with risk factors.
METHODSThe clinic data of 19 patients treated in the Department of Otorhinolaryngology of Guangzhou Children's Hospital between January 2005 to January 2008 were investigated retrospectively. Among them, 5 were < 2 years old, 6 with craniofacial deformity: small mandible and (or) mandibular retrusion (5 cases), transverse facial cleft (1 case), Down's syndrome (2 cases), cerebral palsy (2 cases), chronic bronchitis (3 cases) and mucopolysaccharidoses (1 case). Nineteen patients with symptoms of snoring, mouth breathing, were diagnosed as OSAHS by polysomnography (PSG) and treated by tonsillectomy and (or) adenoidectomy in hospital. All patients were closely followed-up.
RESULTSFourteen patients underwent PSG 6 months to 1 year after operation, 11 patients recovered, the median [percentiles 25; percentiles 75] apnea-hypopnea index (AHI) decreased from the pre-operative 22.5 [16.5; 24.3] times/h to 2.0 [1.5; 4.3] times/h, and the lowest oxygen saturation (LSaO(2)) before operation was 0.63, and was higher than 0.92 after operation, 1 case accompanying with chronic bronchitis, the pulmonary hypertension was improved after operation. One case with Down's syndrome was not significantly improved, preoperative AHI and LSaO(2) was 22.4 times/h and 0.67, and after operation was 14.2 and 0.84; 2 cases accepted adenoidectomy only, snoring, mouth breathing reappeared 3 months after operation, pre-operative PSG results showed AHI 24.6 times/h and 26.6 times/h, LSaO(2) was 0.69 and 0.73, after operation the AHI was 10.6 times/h and 8.5 times/h, LSaO(2) was 0.90 and 0.88, the symptoms disappeared after adenotonsillectomy. Five cases did not have PSG because they lived far away in the other cities, their pre-operative PSG showed AHI 16.4 to 26.2 times/h, LSaO(2) was 0.65 to 0.76. One year after operation, these patients were followed-up by telephone, 4 children were significantly improved, 1 case with mandibular symptoms showed no improvement.
CONCLUSIONSFor OSAHS children accompanying with risk factors, if they have adenoid and tonsil hypertrophy, adenotonsillectomy is the major treatment. Because of the existence of risk factors, perioperative risk increased, even the failure of operation. so these patients must be comprehensively assessed before operation. Satisfied results can be achieved by close observation after operation and management of complications as soon as possible.
Adenoidectomy ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Retrospective Studies ; Risk Factors ; Sleep Apnea, Obstructive ; complications ; surgery ; Tonsillectomy
10.Expression and relationship of Ezh2, Runx3 and caspase-3 in endometrial adenocarcinoma.
Zhen-li GUO ; Ke CHEN ; Xiao-qiu WANG ; Wen HU
Chinese Journal of Pathology 2011;40(6):387-391
OBJECTIVETo investigate the role of the expression of Ezh2, Runx3 and caspase-3 proteins and their correlation in the pathogenesis of endometrial carcinoma.
METHODSExpression of Ezh2, Runx3 and caspase-3 proteins was examined by tissue microarray technique and immunohistochemistry (SP method) in 72 cases of endometrial adenocarcinomas, 60 endometrial hyperplasia and 30 normal endometrial tissues.
RESULTSThe positive expression rates of Ezh2, Runx3 and caspase-3 proteins in endometrial adenocarcinomas were 83.3% (60/72), 26.4% (19/72) and 33.3% (24/72), respectively. The positive rate of Ezh2 protein in endometrial carcinomas was higher than that in normal endometrium and endometrial hyperplasia (16.7%, 33.3%, 63.3%;P < 0.05). However, the positive rate of Runx3 in endometrial carcinomas was lower than that in normal endometrium and endometrial hyperplasia (80.0%, 56.7%; P < 0.01). The positive rate of caspase-3 protein in endometrial carcinomas was lower than that in normal endometrium and endometrial hyperplasia (86.7%, 73.3%, 63.3%; P < 0.01). Positive expression of Ezh2 and Runx3 was related to the histological grade, FIGO stage, and depth of invasion of endometrial adenocarcinomas (P < 0.05), but it was not related to the lymph node metastasis (P > 0.05). Positive expression of caspase-3 protein was related to the histological grade (P < 0.05), but it was not related to the FIGO stage, depth of invasion and the lymph node metastasis of endometrial adenocarcinomas (P > 0.05). The expression of Ezh2 protein was negatively correlated to that of Runx3 (r(s) = -0.262, P < 0.05).
CONCLUSIONSAbnormal expression of Ezh2, Runx3 and caspase-3 proteins is associated with the development and progression of endometrioid adenocarcinoma. Combined analysis of Ezh2, Runx3 and caspase-3 may offer prognostic information for patients with endometrial cancer.
Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Caspase 3 ; metabolism ; Core Binding Factor Alpha 3 Subunit ; metabolism ; DNA-Binding Proteins ; metabolism ; Endometrial Hyperplasia ; metabolism ; pathology ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; metabolism ; Enhancer of Zeste Homolog 2 Protein ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Grading ; Neoplasm Invasiveness ; Neoplasm Staging ; Polycomb Repressive Complex 2 ; Transcription Factors ; metabolism