2.Effects of inducible hypothermia on expressions of PGC-1 and SHP in rat model of hemorrhagic shock
Cheng ZHANG ; Guangrong GAO ; Da LI ; Yongqi SAN ; Xuefeng ZHANG
Chinese Journal of Emergency Medicine 2017;26(2):186-190
Objective To investigate the effects of inducible hypothermia on expressions of peroxisome proliferator activated receptor gamma coactivator-1 (PGC-1) and small heterodimer partner (SHP) in rat model of hemorrhagic shock.Methods SD rats were randomly (random number) divided into three groups:control (C),normothermia (N) and hypothermia (H).Exsanguination was carried out in rats by continuously drawing out venous blood (25 mL/kg) over 15 minutes to establish hemorrhagic shock model.Then,rectal temperatures of rats were maintained by body surface cooling to 32℃ in H group and by body surface warming to 38℃ in N group,respectively.After a shock period of 60 minutes,rats received the infusion of whole blood of their own and lactated Ringer's solution (1 ∶ 2) treatment for 60 min.Rats were warmed to 38℃ by body surface warming and monitored for 3 h after resuscitation.Hematocrit (Hct),base excess (BE),lactate (Lac),and glucose (Glu) were recorded before modeling and after different lengths of hemorrhagic shock period (HSP).The expressions of PGC-1 mRNA and SHP mRNA and the levels of their protein in liver were tested by real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blotting,respectively.Results The H group had lower lactate levels and higher BElevels than the N group [(6.3±2.1) vs.(10.4±1.5) and (-4.7±2.5) vs.(-9.0±3.2)] (P< 0.05).At 72 hours after modeling,there were four survivors in the N group and seven survivors in the H group (P < 0.05,Log Rank).The expressions of PGC-1 mRNA and SHP mRNA increased in N group.Hypothermia resuscitation down-regulated PGC-1 mRNA expression,meanwhile,increased expression of SHP mRNA.Both Hypothermia and Normothermia resuscitation increased SHP protein levels,but decreased PGC-1 protein levels.Conclusions Inducible hypothermia ameliorated acidosis and energy metabolism imbalance through adaptive regulation in PGC-1 and SHP.
3.Expression of Tenascin during Heart Development in Rats
xue-ying, CHENG ; jun, LI ; da-wei, WANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To detect the expression of tenascin-C(TN-C)and tenascin-X(TN-X)in the embryonic heart of rats,investigate their role during the heart development.Methods Sixty-four Spraque-Dawley female rats and 20 male rats were put into the same cage,when vaginal suppository was found,the femate was pregnant 0.5 day(ED 0.5),then executed the pregnant rats on ED 12.5,ED 15.5 and ED 19.5,got out the hearts.Semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR)was used to detect the TN-C and TN-X mRNA expression in the rat hearts aged embryonic day 12.5(ED 12.5),ED 15.5 and ED 19.5.The protein expression of TN-C and TN-X in the embryonic hearts was detected by immunohistochemistry.Results TN-C mRNA was highly expressed in ED 12.5 embryonic hearts(0.683 7?0.043 1),the expression of TN-C mRNA of ED 15.5(0.454 3?0.038 8)and ED 19.5(0.250 5?0.038 4)embryonic hearts decreased,they were different from the expression of ED 12.5(Pa
4.Culture of pancreatic duct-derived stem cells of rats in vitro in the dynamic three-dimension cell culture system
Xuchun CHEN ; Ying CHENG ; Guichen LI ; Rui SHI ; Xiaohang LI ; Da LI ; Yongfeng LIU
Chinese Journal of Organ Transplantation 2012;33(6):371-375
Objective To isolate,purify and identify pancreatic duct derived stem cells (PDSCs) from the pancreatic duct of rats,and culture in the three-dimension cell culture system.Methods Adult male Wistar rats underwent perfusion with collagenase V via the pancreatic duct,then the pancreas was surgically procured,digested,followed by discontinued density gradient centrifuge to isolate ductal tissue from islets.The acinar and ductal tissue was cultivated in serumcontaining medium in the three-dimension cell culture to obtain adherent cells,as PDSCs,which were expanded by consecutive passages.The morphology and characterization of PDSCs on phenotype were examined.Results PDSCs could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three dimension cell culture system.Morphologically,PDSCs had remarkable size,most with one nucleus.PDSCs grew in many layers in three-dimension cell culture system.PDSCs was revealed to express CD29,CD73,CD90,CD105,but not CD14,CD19,CD34,CD45 by FACS,in agreement with MSCs.Conclusion PDSCs of rats could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three-dimension cell culture system.PDSCs lines were successfully established.
5.Correlation study between ~(18)F-FDG uptake and hypoxia inducible factor-1α level,microvessel density in human gliomas
Li, CAI ; Chuan, ZHANG ; Yan-sheng, LI ; Da-cheng, LI ; Xi-ling, XING ; Shuo, GAO
Chinese Journal of Nuclear Medicine 2010;30(1):10-14
Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P<0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P<0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P<0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.
6.Epithelial to mesenchymal transition in the progression of tubulointerstitial fibrosis.
Chinese Medical Journal 2007;120(21):1925-1930
OBJECTIVETo review the mechanisms of epithelial to mesenchymal transition (EMT) and its role in the progression of tubulointerstitial fibrosis.
DATA SOURCESThe data used in this review were obtained mainly from the studies of EMT reported from 2000-2006.
STUDY SELECTIONRelevant articles on studies of EMT in tubulointerstitial fibrosis were selected. Data were mainly extracted from the 45 articles listed in the reference section of this review.
RESULTSThe process of EMT has gained wide recognition as candidate mechanism in progression of chronic fibrotic disorders. New markers were identified and facilitate the observation of EMT. EMT is regulated by many factors through activation of kinase-dependent signaling cascades. Recent findings suggest that EMT is a reversible process, which can be controlled by factors for their epithelial inducing activities.
CONCLUSIONRemarkable progresses of EMT research have been made recently. Preventing or reversing EMT is a promising strategy against renal fibrosis.
Animals ; Connective Tissue Growth Factor ; Disease Progression ; Epithelium ; metabolism ; pathology ; Fibrosis ; Humans ; Immediate-Early Proteins ; metabolism ; Intercellular Signaling Peptides and Proteins ; metabolism ; Mesoderm ; metabolism ; pathology ; Nephritis, Interstitial ; metabolism ; pathology ; Transforming Growth Factors ; metabolism
7.Essentials of pharmacophylogeny: knowledge pedigree, epistemology and paradigm shift.
Da-cheng HAO ; Pei-gen XIAO ; Li-wei LIU ; Yong PENG ; Chun-nian HE
China Journal of Chinese Materia Medica 2015;40(17):3335-3342
Chinese materia medica resource (CMM resource) is the foundation of the development of traditional Chinese medicine. In the study of sustainable utilization of CMM resource, adopting innovative theory and method to find new CMM resource is one of hotspots and always highlighted. Pharmacophylogeny interrogates the phylogenetic relationship of medicinal organisms (especially medicinal plants), as well as the intrinsic correlation of morphological taxonomy, molecular phylogeny, chemical constituents, and therapeutic efficacy (ethnopharmacology and pharmacological activity). This new discipline may have the power to change the way we utilize medicinal plant resources and develop plant-based drugs. Phylogenomics is the crossing of evolutionary biology and genomics, in which genome data are utilized for evolutionary reconstructions. Phylogenomics can be integrated into the flow chart of drug discovery and development, and extends the field of pharmacophylogeny at the omic level, thus the concept of pharmacophylogenomics could be redefined in the context of plant pharmaceutical resources. This contribution gives a brief discourse of knowledge pedigree of pharmacophylogeny, epistemology and paradigm shift, highlighting the theoretical and practical values of pharmacophylogenomics. Many medicinally important tribes and genera, such as Clematis, Pulsatilla, Anemone, Cimicifugeae, Nigella, Delphinieae, Adonideae, Aquilegia, Thalictrum, and Coptis, belong to Ranunculaceae family. Compared to other plant families, Ranunculaceae has the most species that are recorded in China Pharmacopoeia (CP) 2010. However, many Ranunculaceae species, e. g., those that are closely related to CP species, as well as those endemic to China, have not been investigated in depth, and their phylogenetic relationship and potential in medicinal use remain elusive. As such, it is proposed to select Ranunculaceae to exemplify the utility of pharmacophylogenomics and to elaborate the new concept empirically. It is argued that phylogenetic and evolutionary relationship of medicinally important tribes and genera within Ranunculaceae could be elucidated at the genomic, transcriptomic, and metabolomic levels, from which the intrinsic correlation between medicinal plant genotype and metabolic phenotype, and between genetic diversity and chemodivesity of closely related taxa, could be revealed. This proof-of-concept study regards pharmacophylogenomics as the updated version of pharmacophylogeny and would enrich the intension and spread the extension of pharmacophylogeny. The interdisciplinary knowledge and techniques will be integrated in the proposed study to promote development of CMM resource discipline and to boost sustainable development of Chinese medicinal plant resources.
China
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Knowledge
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Medicine, Chinese Traditional
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Phylogeny
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Plants, Medicinal
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chemistry
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classification
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genetics
8.Correlation study on virtual touch tissue quantification and the pathological stages of liver fibrosis in chronic hepatitis B
Meina HAO ; Yuzhen ZHAO ; Cheng LI ; Anlin MA ; Tailing WANG ; Jiping DA ; Rui SHU
Chinese Journal of Ultrasonography 2013;22(9):792-795
Objective To investigate the correlation between virtual touch tissue quantification (VTQ) and the pathological grading of liver fibrosis in chronic hepatitis B.Methods 64 chronic hepatitis B patients (the chronic hepatitis group) and 40 healthy volunteers (the controlled group) were collected.The patients in the chronic hepatitis group were underwent liver biopsy.According to the hepatic fibrosis degree,the patients in the test group were classified into stage 0,1,2,3 and 4.The liver shear wave velocities (SWV) of all the participant were measured by VTQ.The cut-off values were determined by an analysis of receiver operating characteristic (ROC) curve.Results The mean SWV was (1.04± 0.13)m/s in the controlled group.The SWV in stages 0,1,2,3,and 4 were (1.17 ± 0.08)m/s,(1.33 ± 0.32)m/s,(1.53 ±0.32) m/s,(2.09 ± 0.54) m/s,(2.18 ± 0.70) m/s,respectively.There was a significantly difference in SWV between the controlled group and the chronic hepatitis group (F =34.97,P =0.00).The SWV were significantly different not only between stages 0-2,and 3,but also between 0-2 and 4 (F =8.87,P =0.00).A positive correlation was observed between the liver fibrosis and the SWV in the chronic hepatitis group (r =0.67,P =0.00).When a cut off value was set at 1.43 m/s,area under ROC curve was 0.875.The sensitivity and specificity were 100 % and 62.5 %.Conclusions SWV has a better correlation with liver fibrosis.VTQ can make an accurate assessment for stage 3 and stage 4 of the chronic hepatitis B.Therefore,VTQ can be used as a noninvasive and reliable diagnostic indicator for chronic hepatitis B.
9.Expressions of CL16/CR6 and CL12/CR4 in first-trimester human trophoblast cells
Yu HUANG ; Da-Jin LI ; Ming-Yan WANG ; Hai-Dong CHENG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(06):-
Objective To investigate the transcription and protein expressions of chemokines CL16, CL12 and their receptors CR6, CR4 in first-trimester human cytotrophoblast cells and human choriocarcinoma cell line JAR. Methods Transcriptions of CR6, CL16, CR4, CL12 in purified first-trimester human trophoblast cells and JAR line were assessed by semi-quantitative RT-PCR, and protein expressions of CR6, CL16, CR4, CL12 were analyzed in primary cultured villous cytotrophoblasts (VCT), extravillous cytotrophoblasts (EVCT), JAR line and placentas by immunostaining. Results CR6 and CR4 were highly transcribed in primary cultured trophoblast cells with mRNA relative level of 1.12?0.25 and 1.08?0.11 respectively, and their ligands CL16 and CL12 were transcribed moderately with mRNA relative level of 0.89?0.11 and 0.78?0.10 respectively. It was demonstrated that CL16, CL12, CR6 and CR4 were expressed in primary cultured VCT, EVCT, JAR line and placentas by immunostaining. Conclusion The co-expression of CL16/CR6 and CL12/CR4 in trophoblast cells may play a role in the proliferation and differentiation of first-trimester trophoblast cells in a manner of autocrine.
10.Transcription of chemokine receptors in villi and trophoblasts of human first trimes-ter gestation
Xia WU ; Da-Jin LI ; Min-Min YUAN ; Ming-Yan WANG ; Haidong CHENG ;
Chinese Journal of Immunology 1985;0(01):-
Objective: To investigate the mRNA expression of chemokine receptors in human villi and trophoblasts of first trimester gestation . Methods: The authors first obtained villous tissues from fifteen women who had undergone selective termination at 5 - 10 weeks of normal gestation. Total RNA was then extracted, using the TRIzol reagent, from villous tissues or Percoll-gradient purified trophoblasts. Consequently, the expressions of chemokine receptors in villous tissues and trophoblasts were investigated by way of semi-quantitative reverse transcriptase-polymerase chain reaction.Results: The chemokine receptors, CXCR4 and CXCR6, were highly expressed in each villous tissue, while the CCR6, CCR7, XCR1 and CX3CR1 were moderately expressed in villi. The chemokine receptors, CCR1- CCR5, CCR8 - CCR10, CXCR1 -CXCR3, were expressed only in some villous samples, while no CXCR5 mRNA was found in any villous tissue. The authors also found that the freshly isolated and Percoll-purified trophoblasts expressed CCR1, CCR3 - CCR5, CCR8 - CCR9, CXCR1 - CXCR4, CXCR6, XCR1 and CX3CR1 mRNA. Conclusion: A variety of chemokine receptors were expressed in villous tissues and trophoblasts of human first trimester gestation, hence, these receptors may play an important biological role at the materno-fetal interface in normal human pregnancy.