1.Inhibition and mechanism of 15 (S)-hydroxyeicosate traenoic acid on proliferation of hypoxic retinal microvascular endothelial cells in vitro
Xiayun, HU ; Tao, HE ; Yiqiao, XING ; Qiong, WANG
Chinese Journal of Experimental Ophthalmology 2014;32(12):1067-1073
Background Retninal neovascular diseases caused by hypoxia has become a major blinding disease,which is lack of effective chemical treatment currently,it's important to study the molecuar mechanism of the disease,so as to guide the clinical medication.Objective This study was to explore the effect of 15 (S)-hydroxyeicosate traenoic acid (15-HETE) on the proliferation of hypoxic retinal microvascular endothelial cells (RMVECs) and its probable mechanism.Methods RMVECs were isolated from C57BL/6J mice and incubated and then identified with anti-Ⅷ factor antibody by immunochemistry and immunofluorescence.The cells were divided into the normoxia group and the hypoxia group.The hypoxia cell models were established by treated with 125 μmol/L CoCl2.The cells were cultured with serum-free DMEM containing endothelial cell growth supplement (ECGS)and high glucose for 48 hours,and then different concentrations of 15-HETE (0.0,0.1,1.0,5.0 μmol/L) were added in the medium for 48 hours respectively to subgroup the groups.The proliferation of the cells (absorbance,A) was detected using MTT.The relative expression levels of protein and mRNA of hypoxia inducible factor-1α (HIF-1α),bcl-2 and caspase-3 were assayed by reverse transcription PCR (RT-RCR)and Western blot.Results The cells showed the positive response for anti-Ⅷ factor antibody with the positive rate of (94.38 ±4.25)%.No significant difference was found in the cell proliferation of various groups under the normoxia condition (F =0.283,P =0.837),but under the hypoxia condition,the proliferation values were significantly different among various groups (F =702.582,P<0.001).The cell proliferation value in the 1.0 μmol/L 15-HETE group and 5.0 μmol/L 15-HETE group was lower than that of the simple hypoxia group respectively(both at P<0.05).The inhibitory rates in the 0.1,1.0,5.0 μ mol/L 15-HETE groups were (1.09±0.31) %,(21.09± 3.53) % and (49.86 ±4.15) %,showing a dosedependent manner.No significant difference was seen in the expression levels of bcl-2,caspase-3 and HIF-1α mRNA in various groups under the normoxia conditions.However,compared with the simple normoxia group,the relative expressions of bcl-2 mRNA and HIF-1α mRNA in the cells were increased by 1.53 folds and 1.7 folds in the simple hypoxia group respectively,and caspasse-3 mRNA expression decreased by 70% (all at P < 0.05).Under the normoxia condition,the expression of bcl-2 and pro-caspase-3 protein in the cells were not significantly different among the various groups (P>0.05),however,the expressions of bcl-2 and pro-caspase-3 proteins were elevated by 1.6 folds and 1.9 folds in the hypoxia group in compared with the normoxia group (P<0.05).Compared with the simple hypoxia group,the expressions of bcl-2 and pro-caspase-3 were lowed by 40.4% and 42.5% in the 5.0 μmol/L 15-HETE group (P<0.05).Conclusions 15-HETE inhibits the proliferation of RMVECs and therefore suppresses neovascularization by down-regulating the expressions of HIF-1α and bcl-2 and the activation of caspase-3 in a dose-dependent manner.
2.Detection of CALR mutations in peripheral blood of myeloproliferative neoplasm patients with high resolution melting curve analysis
Wenhui WANG ; Yiqiao DU ; Weihua YANG ; Yingdi DONG ; Zhenhua YANG
Chinese Journal of Laboratory Medicine 2017;40(6):456-459
Objective To establish a rapid, accurate and low-cost screening method for the detection of calreticulin (CALR) mutations in myeloproliferative neoplasms (MPN).Methods Seventy cases diagnosed with MPN were collected from 2012 to 2016. PCR combined with high resolution melting (HRM) analysis were used to screen the CALR mutations, and Sanger sequencing and T-A sequencing were applied to verify the HRM positive samples. CALR wild type DNA, type 1 and type 2 mutant DNA samples were selected and analyzed 4 times/day for 5 days to detected the CVs of Tm (melting temperature) respectively. JAK2 mutations were also analyzed in MPN patients to compare the association between JAK2 and CALR mutations.Results PCR-HRM analysis showed 7 cases (26.9%) and 5 cases (20.8%) patients with CALR mutations were screened out from 26 essential thrombocythaemia (ET) cases and 24 primary myelofibrosis (PMF) cases, but no CALR mutations were found in cases with polycythaemia vera (PV). All mutations were confirmed by direct sequencing or cloning sequencing. The CVs for HRM analysis of CALR wild type DNA, type 1 and type 2 mutant DNA samples were 1.91%,1.59% and 1.43%, respectively.There were 47 cases with JAK2 V617F and 1 case with exon12 mutation. No coexistence of JAK2 mutation and CALR mutations were found in a single sample.Conclusion PCR-HRM can be used for rapid screening of CALR mutation. Subsequent sequencing can be applied for rapid diagnosis of MPN patients in clinical practice.
3.Effect and Mechanisms of Isoflurane on energy metabolism after Ischemia in Isolated Rat Hearts [WT5BZ]
Pengcheng XU ; Nuo XU ; Yiqiao WANG ; Jianwei FAN
Chinese Pharmacological Bulletin 2003;0(12):-
Aim To determine the effects and mechanisms of isoflurane on energy metabolism during ischemia and reperfusion in isolated rat hserts.Methods The rat Langendorff model was used,and isolated per-fused rat hearts were separated into untreated,isoflurane,chelerythrine(PKC inhibitor)plus isoflurane,and chelerythrine groups.All the hearts were subjected to treatment before ischemia,followed by 30 min of ischemia and 60 min of reperfusion.Hemodynamic variables were recorded,and metabolites were measured by high-performance liquid chromatography,and analyzed subcellular localization of PKC isoforms by Western blot analysis.Results The recovery of left ventricular developed pressure after ischemia was(33?8)%,(56?9)%,and(30?5)% in the untreated,isoflurane,and isoflurane with chelerythrine groups respectively.Compared with the untreated hearts,isoflurane significantly improved the recovery of left ventricular developed pressure(P
4.Effect of propofol pretreatment on endoplasmic reticulum stress-mediated apoptosis during focal cerebral ischemia-reperfusion in rats
Weixiang TANG ; Likui WANG ; Yiqiao WANG ; Xiuli YANG ; Zhixin GAO ; Yujun SHEN ; Yuanhai LI
Chinese Journal of Anesthesiology 2012;(12):1488-1490
Objective To evaluate the effect of propofol on the endoplasmic reticulum stress-mediated apoptosis during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Eighty adult male Sprague-Dawley rats,weighing 240-280 g,were randomly divided into 4 groups (n =20 each):shame operation group (group S) ; focal cerebral I/R group (group FCIR); propofol pretreatment group (group P); intralipid pretreatment group (group Ⅰ).Focal cerebral I/R was induced by 4 h middle cerebral artery occlusion followed by reperfusion.Propofol was infused at a rate of 12 mg· kg-1 · h-1 starting from 30 min before ischemia until 15 min of ischemia in group P,while intralipid was given instead of propofol in group I.Neurological deficit scores (NDSs) were measured at 6 h of reperfusion in 10 rats chosen from each group and the rats were then sacrificed.Their left brains were removed for determination of brain water content.The left 10 rats were sacrificed and their brains were immediately removed for determination of the expression of C/EBP homologous protein (CHOP),Bcl-2,and activated caspase-3 in the left hippocampi by Western blot.Results Compared with group S,NDSs and brain water content were significantly increased,the expression of CHOP and activated caspase-3 was up-regulated,and the expression of Bcl-2 was down-regulated in group FCIR,NDSs was increased in group P (P < 0.05).Compared with group FCIR,NDSs and brain water content were significantly decreased,the expression of CHOP and activated caspase-3 was down-regulated,and the expression of Bcl-2 was up-regulated in group P,and no significant change was found in each parameter in group Ⅰ (P > 0.05).Conclusion Propofol can reduce focal cerebral I/R injury through inhibition of the endoplasmic reticulum stress-mediated apoptosis in rats.
5.Application of BIS and muscle relaxation monitors in robot-assisted laparoscopic radical prostatectomy
Lina HAO ; Ke CHEN ; Xianfu LU ; Jun LI ; Yiqiao WANG ; Yuanhai LI
The Journal of Clinical Anesthesiology 2017;33(3):248-251
Objective To evaluate the effect of bispectral index (BIS) and muscle relaxation monitoring on robot-assisted laparoscopic radical prostatectomy in elderly patients.Methods One hundred elderly patients (aged 65-80 years,ASA Ⅰ or Ⅱ) who underwent robot-assisted laparoscopic radical prostatectomy were randomly allocated into BIS and muscle relaxation monitoring group (group AA,n=50) and control group (group AC,n=50).In group AA,propofol was infused to achieve the BIS value of 45-55,and we monitored the muscle relaxation to conduct closed-loop infusion of cisatracurium.In group AC,we regulated the depth of anesthetic with the patients` vital signs according to anesthetists` experience.Mean arterial pressures (MAP),heart rates (HR),airway platform pressure (Pplat),and airway peak pressure (Ppeak) were recorded at following time points: before anesthesia induction (T0),after anesthesia induction (T1),10 min (T2),60 min (T3) after artificial pneumoperitoneum,and the end of operation (T4).We recorded dosage of propofol,cisatracurium,sufentanil,remifentanil,vasoactive agent,extubation time and PACU stay time.Results At T1,T2 and T4,the MAP and HR in group AC were significantly higher than those in group AA (P<0.05);at T3,MAP in group AC were apparently lower than those in group AA (P<0.05).Compared with T0,MAP and HR in group AC were significantly increased at T1,T2 and T4(P<0.05),MAP in group AC were obviously reduced at T3 (P<0.05),MAP and HR in group AC were also fluctuated obviously at different time points.MAP and HR in group AA at each point had no statistically significant difference.Compared with T1,Pplat and Ppeak in the two groups were significantly increased at T2-T4 (P<0.05).Pplat and Ppeak in grpup AC were higher than those in group AA at T2,T3 (P<0.05).Compared with group AC,the dosages of propofol and cisatracurium were less in group AA.The postoperative extubation time and PACU stay time were shorter in group AA.Conclusion BIS and muscle relaxation monitoring in robot-assisted laparoscopic radical prostatectomy can effectively stablize hemodynamics,reduce airway pressure fluctuation and the dosage of anesthetics.It also shortens the extubation time and the PACU stay time and improves the anesthesia recovery quality.
6.Relationship analysis between health expenditure and GDP based on health outcomes
Xiaowei MAN ; Yan JIANG ; Liying ZHAO ; Yiqiao WANG ; Baolin HONG ; Xuan ZHAO ; Wei CHENG
Chinese Journal of Health Policy 2016;9(7):66-73
Objective:To put forward some suggestions for the national health development in the filed of health expenditure indicators .Methods:Two health expenditure indicators and four health outcomes indicators were chosen from WB and WHO official websites , and researches on the relationship between health expenditure and outcomes were conducted .We put forward proposed value of the health cost indicator .Results: With different levels of GDP per capita , the scatter diagram of health expenditure and GDP per capita had different rules .When health outcomes were taken into consideration , the rules were the same .When GDP per capita was at different level , better health outcomes were not related to higher health expenditure .Input-output ratio should be taken into consideration fully . Suggestions:The study should be dynamic as GDP develops .More factors can be added in the research , if the data of the factors is available .In this study , the GDP per capita was divided into 4 sections .The division standard of the GDP per capita can be more delicate .What is more important for the development of health expenditure is not contin-uing input , but better input-output ratio because of diminishing of marginal returns .One set of standards cannot be a-dapted to all countries and districts .We should give full consideration to the improvement of people's health instead of increasing input because the ultimate goal is the former .
7.Comparative analysis on the effective competence of clinical head nurses'in Macau and Guangzhou
Sichen WANG ; Liang LU ; Mingxia ZHU ; Yiqiao YIN ; Xiaoxia HU ; Huilan HUO ; Wanhua CHEN
Chinese Journal of Practical Nursing 2013;(2):51-53
Objective To explore the clinical head nurses'effective competence through interviewing nursing supervisors by in depth interview method and to provide reference for constructing training content for clinical head nurses in Macau and Guangzhou.Methods In depth interview was carried out among 8 nursing supervisors from 2 hospitals in Macau and 9 nursing supervisors from 2 tertiary hospitals in Guangzhou.Content analysis was utilized through organizing records and then forming written documentations.Results The themes were drawn after the interview,which were good moral character accomplishment,good communication skill,prospective and creative thinking,favorable executive ability.Forwardlooking and innovative were significantly different in Macau and Guangzhou.Conclusions The effective clinical head nurses' competence is multi-faceted,the results can provide reference for selection,evaluation and training for head nurses.
8.Effects of low-dose low molecular weight heparin on lipopolysaccharide-induced acute lung injury
Yu LIU ; Shaozheng LIU ; Xianfeng WANG ; Yiqiao WANG
Chongqing Medicine 2017;46(26):3608-3610
Objective To explore the effect of low-dose low molecular weight heparin(LMWH) on acute lung injury(ALI) induced by lipopolysaccharide(LPS).Methods Thirty-six male SD rats were divided into 3 groups:normal control group(A),LPS group(B) and LPS plus LMWH group(C),12 cases in each group.ALI was induced by intraperitoneal injection of LPS 6 mg/kg in group B,C.The group C accepted intraperitoneal injection of LMWH 100 U/kg,the groups B accepted intraperitoneal injection of the same volume of normal saline.The animals were killed after 6 h,the pathological changes of the lung were observed under light microscope.Arterial blood gases,lung wet to dry ratio (W/D) and protein content in BALF were detected;the levels of MPO and MDA in lung tissue were determined.The levels of serum interleukin1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α) were determined by ELISA method.Results The PaO2 and pH values in the group B and C were significantly lower than those in the group A,and which in the group C were markedly increased compared with those in the group B(P<0.05);the lung W/D ratio,protein content in BALF,MDA and MPO levels in the lung tissue in the group B and C were significantly higher than those in the group A(P<0.01),while the lung W/D ratio,protein content in BALF,MDA and MPO levels in the lung tissue in the group C were significantly decreased compared with the group B (P<0.05).The levels of plasma TNF-α,IL-1β and IL-6 levels in the group B and C were markedly increased compared with the group A,while which in the group C were significantly decreased compared with those in the group B (P<0.01).Conclusion The LMWH treatment might attenuate LPS-induced acute lung injury.
9.Novel nano-scaled Mn-metal-organic framework for enhancing photodynamic therapy through overcoming tumor hypoxia
Huayuan CUI ; Dan YAO ; Yuxiang WANG ; Yiqiao HU
Journal of China Pharmaceutical University 2019;50(6):678-685
A Mn-clusters-porphyrin metal-organic framework nanosheet(nMn-MOF)was synthesized by coordination chelation to enhance photodynamic therapy. The nanosheet was characterized by dynamic light scattering, transmission electron microscopy and X-ray photoelectron spectroscopy. Oxygen sensor and ICG were used to investigate the production of oxygen and the singlet oxygen(1O2)generation. The cytotoxicity of the nanosheet against tumor cells were detected by CCK-8 assay, and the anti-hypoxia and oxygen-generation ability of nanosheets were investigated by fluorescence staining assay. The results indicated that this nanosheet could catalyze the intracellular H2O2 into O2, which overcame the tumor hypoxia. Furthermore, the generated oxygen was converted to cytotoxic 1O2 under the near infrared light irradiation, thereby enhancing photodynamic therapy.
10.An exploration of deaf people's needs for smooth communication with doctors and nurses
Yuli ZANG ; Yulin WANG ; Chunlan TONG ; Jing LI ; Nan ZHU ; Yiqiao SHEN ; Kuichang YUAN ; Xiaoqin MENG ; Qiang CHENG ; Fenglan LOU
Chinese Journal of Practical Nursing 2008;24(26):67-69
Objective This study was to explore deaf people's needs for a smooth communication with doctors and/or nurses during health consultation in the outpatient department or when being hospitalized and supply reference for compilation of sign language textbooks.Methods Purposive sampling was used and semi-structured interviews were conducted.Six deaf participants who met the inclusion and exclusion criteria were interviewed.It lased for 20 to 40 minutes for each interview.Interviews were recorded digitally and then transcribed and validated.Transcripts were analyzed using a method of thematic analysis.Results Three themes were identified,i.e.needs for sign language interpreters,needs for information and knowledge.and needs for psychosocial and emotional support.Needs for sign language interpreters meant that doctors and nurses were able to communicate with them using simple sign language instead of terminology.The required information and knowledge covered three aspects,i.e.health condition,pharmaceutical therapy,and self-care activity.Conclusions There may exist serious communicative barriers for deaf people during their health consultation in the outpatient department or when being hospitalized.There is a need for healthcare professionals who are competent in sign language to establish the health delivery environment with minimal barriers.