1.Hypoxia-inducible factor-1alpha gene-modified umbilical cord blood mesenchymal stem cells transplantation in the treatment of spinal cord ischemia-reperfusion injury
Chinese Journal of Tissue Engineering Research 2015;(10):1592-1596
BACKGROUND:There are no available therapies for spinal cord ischemia-reperfusion injury, and stem cel transplantation is a focused topics. OBJECTIVE:To observe the therapeutic effect of hypoxia-inducible facotr-1α gene-modified umbilical cord blood mesenchymal stem cels (UC-MSCs) transplantedvia the infrarenal abdominal aorta on spinal cord ischemia-reperfusion injury in rats. METHODS:Thirty adult female Sprague-Dawley rats were randomly divided into three groups, with 10 rats in each group. The infrarenal abdominal aorta of rats was occluded surgicaly for 1 hour, and then the spinal cord reperfusion was restored. At 2 hours after reperfusion, 1 mL of 10% PBS, UC-MSCs suspension and hypoxia-inducible factor-1α-modified UC-MSCs suspension was injectedvia the infrarenal abdominal aorta, respectively, in the three groups. At 1, 6, 12 days after injection, Basso-Beattie-Bresnahan scores were recorded and western blot assay was used to detect hypoxia-inducible factor-1α protein expression in the spinal cord. The motor-evoked potential was determined at 12 days after injection. RESULTS AND CONCLUSION: Compared with the control group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P< 0 .05) in the untransfected group and transfection group. Compared with the untransfected group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P < 0 .05) in the transfection group. Above al, umbilical cord blood mesenchymal stem cel transplantation modified by hypoxia-inducible factor 1α has better effects on spinal cord ischemia-reperfusion injury.
2.Effects of local mild hypothermia on the expression of EMAP-Ⅱ and proEMAP-Ⅱ after cerebral ischemia and reperfusion
Qian SHI ; Shurong DUAN ; Ruihong SUN
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(2):86-90
Objective To investigate the effects of local mild hypothermia on the expression of EMAP-Ⅱ and proEMAP-Ⅱ after cerebral ischemia and reperfusion in rats and to explore the possible neuroprotection mechanism of mild hypothermia.Methods Forty-four male Wistar rats were divided randomly into a sham-operation group (Sham),a normothermia group (NT) and a hypothermia group (HT).Middle cerebral artery occlusion was performed using Longa's method,and reperfusion was allowed after 2 hours of occlusion.Mild hypothermia (33.0 ± 0.5)℃ for 6 hours was initiated at the start of reperfusion,followed by rewarming.Brains were harvested after 6,12,24,48 and 72 hours of reperfusion and used for HE staining to evaluate cellular apzoptosis and immunohistochemical staining for detecting the expression EMAP-Ⅱ and proEMAP-Ⅱ.Results The expression of EMAP-Ⅱ and proEMAP-Ⅱ in the ischemic penumbra was significant at 6 hours in the normothermia and hypothermia groups.It peaked at 12 hours in the normothermia group and 24 hours in the hypothermia group,and then decreased gradually.At 72 hours the expression of EMAP-Ⅱ and proEMAP-Ⅱ in the ischemic penumbras was very close to that in the sham group.EMAP-Ⅱ-positive cells were significantly fewer in the hypothermia group than in the normothermia group at all time points.ProEMAP-Ⅱ-positive cells were significantly more numerous in the normothermia group than in the hypothermia group at 6,12 and 24 hours.Conclusions Mild hypothermia (33.0 ± 0.5) ℃ has a valid neuroprotective effect which involves reducing EMAP-Ⅱ and proEMAP-Ⅱ expression in the ischemic penumbra and inhibiting apoptosis and inflammatory reactions after cerebral ischemia and reperfusion,at least in rats.
3.Experimental Study on Effect of Shexiangbaoxin Pills on Morphosis and Function of Left Ventricle after Myocardial Infarction in Rat
Xinping LUO ; Haiming SHI ; Zhiyu ZENG ; Weihu FAN ; Shouyi WANG ; Ruihong DAI ;
Chinese Traditional Patent Medicine 1992;0(07):-
Objective: To investigate the effect of Shexiangbaoxin Pills on morphosis and function of left ventricle after myocardial infarction in rat. Methods: The rat myocardial infarction models induced by ligaturing coronary artery were randomly divided into the myocardial infarction control group, Shexiangbaoxin Pill group and captopril group. The hemodynamic parameters and cardiac functions in 2 or 6 weeks after treatment were determined. Meantime, the morphological parameters of left ventrile (left cardiac chamber size, attenuation proportion of ventricular wall of infarction area, and expansion index of left ventrile), were studied. Results: Shexiangbaoxin Pills could reduce the distention of left ventrile and expansion of infarction area of rat in 6 weeks after infarction. It could obviously improve the contraction function of left ventrile after myocardial infarction. Conclusion: Shexiangbaoxin Pills can improve left ventricular reconstitution after myocardial infarction.
4.Effect of TLR1 ,TLR2,TLR6 in PBMC from patients with ovarian cancer on tumor microenvironment
Juan XU ; Jianfang LOU ; Xinhui SHI ; Xing KE ; Shuping ZHANG ; Meng WU ; Ruihong SUN ; Lei HUANG ; Fang WANG
International Journal of Laboratory Medicine 2018;39(5):537-542
Objective To explore the pathogenesis of ovarian cancer by investigating the function of Toll-like receptor 1 (TLR1),TLR2 and TLR6 in peripheral blood mononuclear cell(PBMC) from patients with ovarian cancer.Methods PBMC,SK OV 3 co culture system and anti-TLR1,anti-TLR2,anti-TLR6 mAb blocking experiment were used to explore the relationship between TLR1,TLR2 or TLR6 signaling and inflammation in ovarian cancer.Quantitative real time PCR was used to measure interleukin(IL)-1β,IL-6,IL-8,and tumor necrosises factor(TNF)-α in the PBMC.MyD88,TRAF6,TANK,NF-κB and P-NFκB were observed by Western blotting.Results In the PBMC and SK-OV-3 coculture system,we found the activation of TLR signaling pathways,including significantly increased MyD88,TRAF6,TANK and P-NF-κB levels following cocultured with SK-OV-3 in PBMC from ovarian cancer patients.PBMC derived from ovarian cancer patients led to a increase in IL-1β,IL-6 and IL-8 mRNA levels after 24 hours of co-incubation with SK-OV-3 (Fold=1.74,Fold=1.92,Fold=1.65,P<0.05),though there was no difference of TNF-a mRNA expression.In contrast to the ovarian cancer patients,coculture of PBMC derived from benign diseases controls and healthy normal controls decreased IL-1β at the mRNA level (Fold=0.71,P<0.05;Fold=0.72,P<0.05),furthermore the expression of MyD88,TRAF6,TANK,P-NF-κB,and NF-κB showed no changes.PBMC which treated with anti-TLR1,anti-TLR2 or-TLR6 mAb could inhibite inflammatory IL-1β (Fold=0.16,Fold=0.31,Fold=0.29,P<0.05) and IL-6 (Fold=0.14,Fold=0.20,Fold=0.28,P<0.05).Conclusion TLR1/TLR2/TLR6 in PBMC of ovarian cancer patients participate in the recognition of the factors.
5.Significance of peripheral perfusion index in early diagnosis and goal-directed therapy of septic shock patients: a prospective single-blind randomized controlled trial
Yuanfeng SHI ; Ruihong YIN ; Yanli WANG ; Jiguang LI ; Xiaobing CHEN ; Yongpeng XIE ; Caihong GU ; Xiuzhen ZOU ; Kexi LIU
Chinese Critical Care Medicine 2017;29(12):1065-1070
Objective To investigate the application of peripheral perfusion index (PPI) in early diagnosis and goal-directed therapy of septic shock, and to provide reference for the early clinical diagnosis and treatment of septic shock. Methods A prospective single-blind randomized controlled trial (RCT) was conducted. Adult patients with sepsis admitted to emergency medical department and intensive care unit (ICU) of the First People's Hospital of Lianyungang City in Jiangsu Province from January 2013 to December 2016 were enrolled. The patients were randomly divided into two groups (n = 46). The PPI group was defined using PPI < 1.4 as diagnosis of septic shock standard, and PPI > 2 as treatment guide target. Control group was defined according to the traditional diagnostic criteria of shock which systolic blood pressure was less than 90 mmHg (1 mmHg = 0.133 kPa) or systolic blood pressure value decrease> 40 mmHg baseline and bundle treatment was performed. The volume of fluid resuscitation, organ dysfunction, the sequential organ failure score (SOFA), acute physiology and chronic health evaluationⅡ (APACHE Ⅱ) score,continuous renal replacement therapy (CRRT) time, mechanical ventilation (MV) time, the length of ICU stay and 28-day mortality were observed. Results There were 39 and 27 septic shock patients in PPI group and control group respectively. The diagnostic criteria of traditional septic shock with blood pressure as "gold standard", the sensitivity of PPI < 1.4 for septic shock was 94.3%, the specificity was 28.2%, the authenticity was 66.3%, the positive predictive value was 64.1%, the negative predictive value was 78.6%, the positive likelihood ratio was 1.31, the negative likelihood ratio was 0.18. The per capita fluid replacement within 24 hours in the PPI group was significantly higher than that in the control group (mL: 4 601±1 250 vs. 3 458±1 006, P < 0.01), but there was no significant difference in the per capita volume of the patients diagnosed as septic shock (mL: 4 596±1 320 vs. 4 205±1 058, P > 0.05). Compared with the control group, the PPI group treated patients within 48 hours with less vascular active drugs (cases: 6 vs. 15), APACHE Ⅱand SOFA score were lower (48 hours: APACHE Ⅱ was 10.2±2.1 vs. 12.0±3.2; 72 hours: SOFA was 5.1±1.8 vs. 6.0±2.1, APACHE Ⅱ was 8.9±1.8 vs. 9.8±2.2), the period of CRRT and the length of ICU stay were shorter [the period of CRRT (days): 3.0±0.9 vs. 3.6±1.4, the length of ICU stay (days): 5.2±2.1 vs. 6.3±2.9), the difference was statistically significant (all P < 0.05). There was no significant difference in the liver and kidney function index, arterial blood lactic acid (Lac), MV time (days: 3.3±1.4 vs. 3.5±1.2) and 28-day mortality (15.22% vs. 19.57%) between two groups (all P > 0.05). Conclusions The inadequacy of microcirculatory perfusion by oximetry-derived PPI is more sensitive to the diagnosis of septic shock than hypotension of systemic circulation. With PPI guiding the fluid resuscitation of septic shock patients, vasopressors can be withdrawn earlier and the duration of the CRRT and ICU can be decreased.