1.MCPIP1 induces cell cycle arrest in breast cancer cell line MDA-MB-231
Wenbao LU ; Mingming LIU ; Ruijuan XIU
Basic & Clinical Medicine 2017;37(5):608-613
Objective To investigate the functions of Monocyte chemotactic protein-induced protein 1 (MCPIP1) in human breast cancer cell line MDA-MB-231.Methods MDA-MB-231 cells were transfected with GFP-tagged MCPIP1 by Tet-on inducing expression system.Endogenous MCPIP1 was knocked down by stable expressing shRNA.MTT assay was performed to measure the growth of MDA-MB-231 cells after overexpression or knockdown of MCPIP1.FACS method was used to analyze cell cycle in MDA-MB-231 cells.Real-time PCR was used to test the expression of cell cycle-related mRNAs expression and their half-lives.RNA-IP experiment was conducted to detect the mRNA directly enriched by MCPIP1.Luciferase assay was performed to determine whether the mRNA decay was mediated through 3′UTR.Results MCPIP1 overexpression significantly inhibited cell proliferation(P<0.05), while knockdown MCPIP1 promoted cell proliferation with statistical significances (P<0.05).MCPIP1 induced cell cycle arrest in MDA-MB-231 with statistical significance (P<0.01).MCPIP1 overexpression reduced the half-lives of cell cycle mRNAs (CDK2,CDK6,cyclin D1,cyclin E1,respectively) with significance (P<0.01).In addition, cell cycle-related mRNAs were able to be pulled down by GFP-MCPIP1 but not isotype IgG(P<0.05).Compared with control vector, MCPIP1 significant suppressed luciferase activities of all four 3′UTR reporters (P<0.05).Conclusions MCPIP1 functions as a tumor suppressor in human breast cancer cell line MDA-MB-231 through inducing G1 cell cycle arrest.
2.The exploration of the value of superior vena cava-jugular vein oxygen saturation-differential value in evaluating brain damage and prognosis of severe traumatic brain injury
Shengqiang YANG ; Wenbao YANG ; Jihua LIU ; Baojun HOU ; Qibiao SHI
Chinese Journal of Postgraduates of Medicine 2014;37(8):11-14
Objective To explore the relationship between superior vena cava-jugular vein oxygen saturation-differential value (ScVO2-SjVO2) and injury severity and prognosis in patients suffered from severe traumatic brain injury,finding out the chnical value.Methods Forty-three patients with severe tranmatic brain injury were selected and divided into two groups according to Glasgow Coma Scale (GCS) score,19 cases were enrolled in GCS 3-5 scores group;24 cases in GCS 6-8 scores group;20 contemporaneous cases in control group had conscious and without traumatic brain injury.The changes of ScVO2-SjVO2 at 0,12,24,48 h after admission of ICU among the groups were observed.In addition according to the degree of change in ScVO2-SjVO2,12 cases were enrolled in ScVO2-SjVO2 < 0.05 group;14 cases in ScVO2-SjVO2 0.05-0.15 group; 17 cases in ScVO2-SjVO2 > 0.15 group.The GCS score and mortality rate in patients after brain injury 28 d among groups were observed.Results GCS 3-5 scores group compared with control group,ScVO2-SjVO2 reduced significantly at 0,12,24,48 h after admission of ICU (-0.071 ±0.032 vs.0.093 ±0.049,-0.097 ±0.052 vs.0.081 ±0.052,-0.035 ±0.031 vs.0.089 ± 0.059,0.036 ± 0.033 vs.0.081 ± 0.064),there were significant differences (P < 0.01).GCS 6-8 scores group compared with control group,ScVO2-SjVO2 dramatically increased at 0,12,24 h after admission of ICU(0.173 ± 0.043,0.158 ± 0.067,0.123 ± 0.073),there were significant differences (P < 0.01 or < 0.05).ScVO2-SjVO2 <0.05 group and ScVO2-SjVO2 >0.15 group compared with ScVO2-SjVO2 0.05-0.15group,the mortality rate significantly increased,the mortality rate were 8/12,4/17,1/14,respectively,there were significant differences (P <0.01).ScVO2-SjVO2 <0.05 group and ScVO2-SjVO2 >0.15 group compared with ScVO2-SjVO2 0.05-0.15 group,GCS score decreased significantly[(4.17 ± 1.34),(5.88 ±1.35) scores vs.(6.86 ± 1.35) scores],and there was significant difference (P <0.01 or <0.05).Conclusions When ScVO2-SjVO2 < 0.05,the difference is negatively related to the deg~e of brain injury.When ScVO2-SjVO2 >0.15,the difference is positively related to the degree of brain injury;ScVO2-SjVO2 can assess injury severity objectively in patients who suffered from severe traumatic brain injury,and it also expresses the rate of cerebral oxygen metabolism.It can provide a basis for treatment accurately and timely,and judgment prognosis.
3.Isolation and identification brain microvessel pericytes in rats
Weiwei QIN ; Wenbao LU ; Shuying LIU ; Hongwei LI ; Ruijuan XIU
International Journal of Cerebrovascular Diseases 2011;19(7):531-534
Objective To explore the method of primary isolation, cultivation and identification of rat brain microvessel pericytes. Methods The brain tissue of 10 3 week-old Wistar rats was separated sterilely. The brain microvessel fragments were separated using two-step enzyme digestion and one-step gradient centrifugation and were seeded in 35-mm dishes for primary culture. The cell morphology was observed by phase contrast microscopy; the immunofluorescence assay was used to identify the associated antigns, such as the α-smooth muscle actin (α-SMA), neuron-glial antigen 2 (NG2), von Willebrand factor (vWF), and glial fibrillary acidic protein (GFAP). Methyl thiazolyl tetrazolium was used to determine the cell growth curve. Results Pericytes climbed out from the adherent brain microvascular fragments around,showing polygonal, and the cell fusion was 95% after 12-14 days. Immunofluorescence staining revealed that the molecular markers of the pericytes α-SMA and NG2 related antigens showed double positive, while the vWF and GFAP related antigens showed double negative and the cultured cells were confirmed as brain microvascular pericytes. The growth rate of primary cells was slower. The passage cells entered into logarithmic growth phase after 36 to 60 hours and entered into plateau phase after 72 to 108 hours. Conclusions This method may successfully isolate rat brain microvascular pericytes with higher purity.
4.Reform on teaching mode of battlefield first aid based on the traits of the medics
Yuexian DENG ; Wenbao LIU ; Lei JIANG ; Jianjun ZHAO ; Chao QIN
Chinese Journal of Medical Education Research 2013;(6):600-602
Medics play a very important role in military medical supporting forces who take on lots of basic work of medical units.It was assigned to us to train the medics for the first time this year in our university.To enhance the teaching effect of the course named first aid in battlefield,a series of measures were taken including modularizing the teaching contents,diversifying the teaching means,dividing students into some groups,strengthening the practice and examinations based on the traits of the medics and so on.At last,some countermeasures were put forward aiming at the process of teaching.
5.Discussion on the Building of Teacher Staff on Post-oriented Education in Army
Yuexian DENG ; Guoliang CHENG ; Chao QIN ; Wenbao LIU ; Lunlun HANG
Chinese Journal of Medical Education Research 2003;0(02):-
With the development of world new military revolution,the objective of education has transformed from degree-oriented to post-oriented in military university.Based on status-on-quo investigation,the author finds some deficiencies in building of teacher staff and put forward some measures.
6.Sequential blood purification effects on cardiac function in patients with acute organophosphorus pesticide poisoning
Zhen LIU ; Shengqiang YANG ; Wenbao YANG ; Jinggang LIU ; Jiyin YUAN ; Qibiao SHI
Chinese Journal of Postgraduates of Medicine 2015;38(7):482-485
Objective To investigate the effects of sequential blood purification on cardiac function in patients with acute organophosphorus pesticide poisoning (AOPP),and to analyze its application value.Methods Forty-five patients with severe AOPP and toxic myocarditis were researched and randomly divided into two groups.Sequential blood purification included hemoperfusion(HP) and continuous veno-venous hemofiltration (CVVH).In treatment group,23 patients were treated with HP + CVVH on the basis of the conventional therapy.Twenty-two patients of control group received the conventional therapy with HP.The differences of two groups on left ventricular ejection fraction (LVEF) and B-type natriuretic peptide (BNP) were compared before treatment and at 24,48 and 72 h after treatment.Results Before treatment,the levels of LVEF and BNP in treatment group were (47.7 ± 9.8)% and (511.3 ± 285.3) ng/L,in control group were (47.3 ± 10.2)% and (535.4 ± 303.9) ng/L,and there were no significant differences (P > 0.05).The levels of LVEF at each time point after treatment in treatment group were significantly higher than those of control group:(52.7 ± 7.3) % vs.(47.8 ± 8.4)%,(55.2 ± 5.2)% vs.(50.6 ± 7.1)%,(60.7 ± 6.3)% vs.(53.3 ± 5.2)%,and the levels of BNP in treatment group were significantly lower than those of control group:(320.7 ± 181.8) ng/L vs.(496.7 ±268.9) ng/L,(252.7 ±113.8) ng/L vs.(384.3 ±167.5) ng/L,(122.4 ±66.7) ng/L vs.(294.8 ± 121.8) ng/L,there were significant differences between two groups (P< 0.05 or < 0.01).In treatment group,the levels of LVEF at 24,48 and 72 h after treatment were significantly increased:(52.7 ± 7.3)%,(55.2 ± 5.2)%,(60.7 ± 6.3)% vs.(47.7 ± 9.8)%,and the levels of BNP were significantly decreased:(320.7 ± 181.8),(252.7 ± 113.8),(122.4 ±66.7) ng/L vs.(511.3 ±285.3) ng/L,there were significant differences (P < 0.01).In control group,the levels of LVEF and BNP at 24 h after treatment were (47.8 ± 8.4)% and (496.7 ± 268.9) ng/L,before treatment were (47.3 ± 10.2)% and (535.4 ± 303.9) ng/L,and there were no significant differences (P > 0.05);the levels of LVEF at 48 and 72 h after treatment were significantly increased:(50.6 ± 7.1)%,(53.3 ± 5.2)% vs.(47.3 ± 10.2)%,the levels of BNP were significantly decreased:(384.3 ± 167.5),(294.8 ± 121.8) ng/L vs.(535.4 ± 303.9) ng/L,and there were significantl differences (P < 0.05 or < 0.01).Conclusion Sequential blood purification treatment of AOPP can reduce myocardial injury and improve the cardiac function.
7.Effects of the β-blockers on cardiac protection and hemodynamics in patients with septic shock:a prospective study
Shengqiang YANG ; Zhen LIU ; Wenbao YANG ; Guizhen ZHANG ; Baojun HOU ; Jihua LIU ; Qibiao SHI
Chinese Critical Care Medicine 2014;(10):714-717
Objective To investigate the effects of β-blockers on cardiac protection and hemodynamic in patients with septic shock. Methods A prospective randomized controlled trial was conducted. Forty-one patients with septic shock in accordance with early goal directed treatment and met the target within 6 hours,and admitted to intensive care unit (ICU)of Affiliated Huxi Hospital of Jining Medical College from January 2012 to January 2014 were enrolled. The patients were divided into treatment group (n=21)and control group (n=20)by random number table. The patients in both groups were given the standard treatment,esmolol was giving to patients in treatment group in order to control the heart rate (HR)below 100 bpm within 2 hours,and the patients in control group only received standard treatment. The changes in hemodynamic parameters〔mean arterial pressure(MAP),central venous pressure(CVP), HR,cardiac index(CI),stroke volume index(SVI),systemic vascular resistance(SVRI),global end diastolic volume index(GEDVI)〕,biochemistry metabolic of tissue〔central venous oxygen saturation(ScvO2),lactic acid(Lac)〕,and cardiac markers 〔troponin I (cTnI)〕before and 12,24,48,72 hours after the treatment were recorded. Results①Before treatment,the hemodynamic parameters,tissue metabolism index and cTnI had no significant differences in both groups (all P>0.05).②The hemodynamic parameters after treatment in the control group showed no significant difference compared with that before treatment. HR and CI in the treatment group were gradually declined after treatment,SVRI and GEDVI were gradually increased. There were significant differences in HR,CI,SVRI,and GEDVI between treatment group and control group from 12 hours on〔HR(bpm):93±4 vs. 118±13,CI (L·min-1·m-2):3.3 ±0.8 vs. 4.5 ±0.6,SVRI (kPa·s·L-1·m-2):159.2 ±27.4 vs. 130.5 ±24.2,GEDVI(mL/m2):668 ±148 vs. 588 ±103,P<0.05 or P<0.01〕. MAP,CVP and SVI in the treatment group showed no significant changes. ③Lac after treatment in both groups was decreased slowly,Lac (mmol/L)at 12 hours after treatment was significantly decreased compared with that before treatment (control group:8.8 ±3.2 vs. 9.8 ±3.4,treatment group:9.5±3.1 vs. 10.5±4.1,both P<0.05). The Lac of control group and treatment group were 2.5±1.2 and 2.7±1.1 at 72 hours after treatment,and there was no significant difference between two groups (all P>0.05). The ScvO2 was not decreased in both groups.④Compared with before treatment,cTnI in the control group was gradually increased,peaked at 72 hours,and that in the treatment group was gradually increased,peaked at 24 hours and then gradually declined. Compared with control group,the cTnI (μg/L)in the treatment group was decreased significantly at 24,48,72 hours (1.15 ±0.57 vs. 1.74 ±0.77,0.93 ±0.52 vs. 2.15 ±1.23,0.52 ±0.36 vs. 2.39 ±1.17,all P<0.01). Conclusionsβ-blockers (esmolol) can improve cardiac function and myocardial compliance,reduce the myocardial injury in patients with sepsis shock. Although β-blockers can decrease cardiac output,it has no influence on the circulation function and tissue perfusion.
8.Establishment of an in vitro blood-brain harrier modal by coculturing brain microvascular endothelial cells and pericytes
Wenbao LU ; Weiwei QIN ; Qiuju ZHANG ; Hongwei LI ; Shuying LIU ; Ruijuan XIU
International Journal of Cerebrovascular Diseases 2012;20(5):338-342
Objective To establish a stable in vitro model of blood-brain barrier (BBB) simulating in vivo state using the primary-cultured rat brain microvascular endothelial cells (BMVECs) and pericytes.Methods The primary rat BMVECs and pericytes were isolated,purified and cultured.The isolated cells were identified by immunocytochemical staining method.An in vitro model of BBB was constructed using Transwell inserts (pore size 0.4 μm) coculture.Its barrier function was evaluated by the 4-hour leakage test,tight junction protein identification,transendothelial resistance detection,and permeability test.The difference between the cocultured model and simple BMVEC model across the membrane resistance values,and the permeability difference of the small molecule sodium fluorescein (Na-F) were compared.Results Confluent BMVEC monolayers demonstrated a typical cobblestone appearance and the pericytes displayed irregular shape and overlapping growth.Immunodouble labeling technique identification showed that the pericytes positively expressed α-srmooth muscle actin (α-SMA) and neuron-glial antigen 2 (NG2); after the fusion of cocultured model endothelial cells,the surface leakage test became positive; immnocytochemical staining shows that a continuous and dense tight junction formed between the endothelial cells; compared to the BMVEC model,the transendothelial electrical resistance of the cocultured model increased significantly (190.762 ± 10.326 Ω/cm2 vs.96.503 ± 8.012 Ω/cm2; t=- 24.489,P <0.01),and the permeability decreased significantly (56.149% ± 3.572% of the single endothelial model; t =19.330,P < 0.01 ).Conclusions The primary isolated rat BMVECs and pericytes cocultured the morphology,structure and barrier function of in vitro model have the basic characteristics of BBB,and they have provided a useful tool for the research of BBB.
9.Clinical application of transnasal high volume oxygen therapy in acute heart failure
Ke ZHANG ; Fenge TIAN ; Jinggang LIU ; Shengqiang YANG ; Wenbao YANG
Chinese Journal of Primary Medicine and Pharmacy 2019;26(5):527-531
Objective To explore the clinical value of transnasal high volume oxygen therapy in the treatment of patients with acute heart failure.Methods From January 2016 to January 2018,61 cases with acute heart failure in Huxi Hospital Affiliated to Jining Medical College were selected.The patients were randomly divided into control group and treatment group according to the digital table,31 cases in the control group and 30 cases in the treatment group.The two groups were routinely given control of fluid volume,analgesia,strong heart,diuresis,vasodilator,anti-platelet aggregation,camp support and so on.The control group was given conventional oxygen therapy,and the treatment group was treated with high flow oxygen through nose.Before treatment and 12h,24h,48h,72h after treatment,the left ventricular ejection fraction (LVEF),oxygen index (PaO2/FiO2),serum lactic acid (Lac),B type sodium and titanium (BNP) in serum,and the application rate of non-invasive mechanical ventilation and invasive mechanical ventilation in 7d were observed in the two groups.Results Compared with the control group,the LVEF of the treatment group in each time point increased [(35.58 ± 3.64) % vs.(37.77 ± 3.76) %,(37.87 ± 3.58) % vs.(40.07 ±3.36)%,(44.94 ±3.19)% vs.(46.83 ±3.21)%,(47.55 ±3.45%)% vs.(40.07 ±3.36%)%,t =-2.308,-2.466,-2.316,-2.487,all P < 0.05].The PaO2/FiO2 of the treatment group increased significantly at each time point after treatment [(177.39 ± 10.62) mmHg vs.(184.17 ± 10.49) mmHg,(188.00 ± 11.72) mmHg vs.(198.57 ± 18.47) mmHg,(204.06 ± 17.69) mmHg vs.(221.40 ± 23.80) mmHg,(265.23 ± 34.51) mmHg vs.(290.37 ± 26.72) mmHg,t =-2.507,-2.678,-3.236,-3.174,all P < 0.05].The BNP level of the treatment group decreased significantly at each time point after treatment [(2 462.90 ± 288.00) ng/mL vs.(2 264.53 ± 366.44) ng/mL,(1 646.61 ± 377.19) ng/mL vs.(1 474.07 ± 214.03) ng/mL,(991.94 ± 242.95) ng/mL vs.(811.90 ±258.67) ng/mL,(653.77 ± 147.671) ng/mL vs.(526.47 ± 127.87) ng/mL,t =2.355,2.187,2.803,3.594,all P < 0.05].The Lac level of the treatment group decreased significantly at 12h and 24h after treatment [(5.05 ± 0.69) mmol/L vs.(4.55 ± 0.80) mmol/L,(3.68 ± 0.89) mmol/L vs.(3.13 ± 0.77) mmol/L,t =2.610,2.601,all P < 0.05],but there were no statistically significant differences between the two groups at 48h and 72h after treatment [(1.62 ± 0.65) mmol/L vs.(1.53 ± 0.65) mmol/L,(1.36 ± 0.64) mmol/L vs.(1.26 ± 0.46) mmol/L,all P > 0.05].In the control group and the treatment group,the incidence rates of non-invasive mechanical ventilation in 7d were 35.48% (11/31),13.33% (4/30),respetively,the difference was statistically significant (x2 =4.034,P < 0.05).In the control group and the treatment group,the incidence rates of invasive mechanical ventilation in 7d were 12.90% (4/31),3.33% (1/30),respetively,the difference was statistically significant (x2 =4.957,P < 0.05).Conclusion Nasal high flow oxygen therapy has better clinical effect on patients with acute heart failure.It is a more active treatment measure,and is worthy of clinical application.
10.The therapeutic effects of Fuyanshu Capsule on chronic endometritis in rats
Pu HUANG ; Jun ZHOU ; Gang CUI ; Baode YANG ; Wenbao ZHAO ; Chuanhao LIU ; Hao HU
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(3):404-407,413
Objective To observe the effects of Fuyanshu Capsule on phenol mucilage-induced endometritis rats and the possible anti-inflammation mechanism of the therapeutic effects.Methods Chronic endometritis in rats was induced by injection of phenol mucilage suspension into the uterus.Sixty female SD rats were randomly divided into 6 groups,namely,sham-operation group (distilled water,10mL/kg),model group,Jinji capsule group (0.65 g/kg),and Fuyanshu Capsule groups (1.8 g/kg,0.9 g/kg and 0.45 g/kg).After the rats were treated 28 days with corresponding medicine by intragastric administration,the pathology of the endometrium and changes of tumor necrosis factor α(TNF-α)and interleukin-2 (IL-2)levels were detected to evaluate the effects of Fuyanshu Capsule. Results Fuyanshu Capsule (1.8 g/kg and 0.9 g/kg)ameliorated the body weight reduction caused by endometritis in rats.Fuyanshu Capsule (1.8 g/kg,0.9 g/kg and 0.45 g/kg)reduced the ratio of the swelling uterus and ovaries to body weight of the rats.It ameliorated obviously the hyperplasia,necrosis and degeneration of endometrial epithelia and infiltration of inflammatory cells.The capsule (1.8 g/kg)decreased the serum IL-2 level in the rats with phenol mucilage-induced endometritis. Conclusion The anti-inflammatory effect of Fuyanshu Capsule on chronic endometritis induced by phenol mucilage in rats may be related to the decrease of IL-2 level.