1.Effect of bone marrow mononuclear cells transplantation on the expression of transforming growth factor beta 1 and connective tissue growth factor in chronic renal damage rats
Chinese Journal of Tissue Engineering Research 2009;13(49):9645-9649
BACKGROUND: The stem cell transplantation achieved progress in treating acute kidney injury. However, cell transplantation in treating chronic renal damage, as well as the protection mechanism, remains poorly understood.OBJECTIVE: To observe the changes of urine protein and renal function, as well as the effects of bone marrow mononuclear cells on expression of transforming growth factor β1 (TGF-β1) and connective tissue growth factor (CTGF) in chronic renal damage rats.DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Experimental Animal Center of General Hospital of Shenyang Military Command from May to October 2008.MATERIALS: Totally 30 female SD rats, with SPF grade, were randomly divided into the control, model and cell transplantation groups, with 10 animals in each group. Additional 10 SD rats were used to prepare bone marrow mononuclear cells.METHODS: Chronic renal damage models were prepared by modified hepatic resection, and 1.0 mL mononuclear cells were injected at 10 weeks after model preparation, with 1×10~8 cells per animal. Rats in the model group were injected same volume of physiological saline.MAIN OUTCOME MEASURES: The 24 hour urinary protein excretion quantity and renal function changes, histopathological change of renal tissues, as well as the expression of TGF-β1 and CTGF.protein excretion, urea nitrogen and creatinine levels (P < 0.01), which especially greater in the model group (P < 0.01 or P
3.Estrogen effects on adipogenic differentiation of cryopreserved adipose-derived mesenchymal stem cells from the adipose capsule of kidney
Hui ZHANG ; Hongguang ZHENG ; Dewei ZHANG ; Yuming MEI
Chinese Journal of Tissue Engineering Research 2013;(27):4998-5004
BACKGROUND: Estrogen exerts a negative regulatory role in adipogenic differentiation of adipose stem cells, but there is no report concerning estrogen effects on adipogenic differentiation of adipose stem cells from the adipose capsule of kidney after long-term cryopreservation. OBJECTIVE: To explore the impact of estrogen on adipogenic differentiation of fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule or those after long-term cryopreservation. METHODS: Passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule were divided into four groups, al of which were induced to adipogenic cells by induced fluid: fresh cells + 10-7 mol/L estrogen, cryopreserved cells + 10-7 mol/L estrogen, fresh cells and cryopreserved cells groups. Oil red O staining and adipogenic quantitative detection were performed at 14 days after induced adipogenic differentiation. RESULTS AND CONCLUSION: There were no differences in the morphology and arrangement between the cryopreserved and fresh cells. Both cryopreserved and fresh cells expressed CD29 and CD 44, but did not express CD31. Intracel ular lipid droplets were observed after adipogenic differentiation by oil red O staining, and the cells were positive for oil red O staining. The adipogenic volume comparison among the four groups was detected on day 14 after adipogenic differentiation, and the absorbance values showed significant difference between the fresh cells and fresh cells + estrogen groups, as wel as between the cryopreserved cells and cryopreserved cells + estrogen groups, but no difference between the fresh and cryopreserved cells groups. It is proved that low-dose estrogen can inhibit the adipogenic differentiation of long-term cryopreserved adipose-derived mesenchymal stem cells from the kidney adipose capsule; however, there is no significant difference between passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule in adipogenic differentiation.
4.Analysis on the Heavy Metal Residues in Codonopsis pilosula in Shanxi Pingshun County
Dewei ZHANG ; Juanjuan HU ; Ruili MENG ; Shibi YE ; Nong ZHOU
China Pharmacy 2016;27(24):3417-3419
OBJECTIVE:To establish a method for the residues determination of Pb,Cd,Cu,As and Hg in Codonopsis pilo-sula,and evaluate the quality evaluation of C. pilosula of Pingshun County in Shanxi province. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry was adopted with KED scanning model,RF power was 1 550 W,sampling depth was 5.0 mm,plasma gas(argon)flow rate was 16.0 L/min,helium partial pressure was 0.1 mbar,argon gas was 0.6 mbar, the vacuum degree of 5×10-8 mbar,branch turbopump speed was 1 000 hz,sampling cone aperture was 1.0 mm,skimmer aperture was 0.5 mm,the spray chamber temperature was 2.7 ℃,the data collection was repeated 3 times. RESULTS:The linear range was 0-20 ng/ml for Pb(r=0.999 3),0-10 ng/ml for Cd(r=0.998 5),0-250 ng/ml for Cu(r=0.998 8),0-20 ng/ml for As(r=0.999 0) and 0-1.0 ng/ml for Hg(r=0.997 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 95.80%-100.20%(RSD=1.85%,n=6),94.50%-98.00%(RSD=1.26%,n=6),98.52%-102.43%(RSD=1.60%,n=6), 94.90%-98.70%(RSD=2.29%,n=6)and 96.00%-101.00%(RSD=1.84%,n=6);the limits of detection were 0.021 0,0.003 4, 0.043 7,0.115 6 and 0.005 6 ng/kg,respectively. Pb,Cd,Cu,and As were detetcted,and Hg was not detected,the range of total contents was 7.185 2~12.558 0 mg/kg. CONCLUSIONS:The method is simple with good precision,stability and reproducibility, and can be used for the residues determination of Pb,Cd,Cu,As and Hg in C. pilosula;heavy metal residues in C. pilosula in Shanxi Pingshun county does not exceed limit values of national and industry standards.
5.Application of carbon nanoparticles in papillary thyroid microcarcinoma surgery
Dewei LI ; Qiang WANG ; Ning LI ; Shuilong ZHANG
Chinese Journal of Endocrine Surgery 2016;10(4):298-302
Objective To explore the significance of carbon nanoparticles in routine central lymph node dissection of papillary thyroid microcarcinoma (PTMC).Methods 272 cases of PTMC admitted from 2013 to 2015 were retrospectively analyzed and they were divided into two groups,the experimental group who were given carbon nanoparticles during surgery (136 cases) and the control group (136 cases) without carbon nanoparticles.The total number of central lymph nodes,number of transfer,and the black dye lymph node number and transfer number in the experimental group were recorded.The total number of lymph nodes,and number of transfer in the control group were recorded.The metastasis rate of the two groups were analyzed.The number of parathyroid mistakenly cut and hypocalcemia cases of the two groups were counted.Parathyroid function was observed by determination of Ca2+ and PTH in blood.The incidence of recurrent laryngeal nerve (RLN) injury of the two groups was compared.Results The total number of central lymph nodes was 1216 and the transfer number was 481 in the experimental group,higher than those of the control group.The positive lymph node rate of the two groups was 39.6% and 25.9% respectively,and the difference was statistically significant.The mumber of patients with central lymph node metastasis was 63 and 42 respectively for the experimental group and the control group,and the transfer rate was 46.3% and 30.8% respectively.The difference was statistically significant.The rate of parathyroid mistakenly cut was 2.6%(7/261) and 14.7% (28/193) respectively for the experimental group and the control group,and the difference was statistically significant.The difference of Ca2+ and PTH value at 3h,6h,and 12h after surgery between the two group was statistically significant.No RLN injury occured.5 cases in the experimental and 8 cases in the control group had temporary RLN injury.The difference was not statistically significant.Conclusions The application of carbon nanoparticles in PTMC surgery can help to improve the thoroughness of central lymph node dissection and to protect parathyroid function.However,its benefits to protect the recurrent laryngeal is uncertain.
6.The feasibility study of repairment of articular cartilage defects with bone marrow-derived mesenchymal stem cells seeded on acellular amniotic membrane
Hao CHEN ; Zhijin ZHANG ; Dewei ZHAO ; Lin GUO
Chinese Journal of Microsurgery 2014;37(3):254-257
Objective To study the effect of repairment of articular cartilage defects in non weight-bearing area of rabbit with bone marrow-derived mesenchymal stem cells (BMSCs) seeded on human acellular amniotic membrane (HAAM).Methods From July 2012 to March 2013,bone marrow-derived mesenchymal stem cells were isolated and purified from rabbit in vitro.The cells were seeded on human acellular anniotic membrane at the concentration of 1.63 × l05/cm2.From 7 days to 8 days after cultured,the complexes of BMSC and HAAM were examined under electronmicroscope,light microscope and by HE stain.Full thickness empty defects measuring 4 mm in diameter by 3 mm depth were prepared in femoral intercondylar fossa of 24 rabbits.The rabbits were randomized into two groups:group A and group B with 12 each group.The defects of right knees were served as control and the left as experimental group.BMSCs/HAAM composite was cultured and then transplanted into the defect of left knee joint in group A as group BMSCs/HAAM and HAAM into group B as group HAAM.These rabbits were killed at 4 and 12 weeks after surgery in each group and the newly cartilage samples were evaluated grossly,histologically are graded.Results In the 4th and 12th week after the operation,the regenerated tissue were white,soft and smooth.Chondrocytes were found in the tissue In the 12th week,the morphology,distribution and arrangement of the regenerated tissues were similar to normal cartilage in the knees with HAAM-BMSCs transplantation.The regenerated tissues grew to be integrated with the surrounding normal cartilage with obscure boundary between them.Chondrocytes were found in all layer of the tissue,surrounding normal cartilage with obscure boundary between them.In the HAAM transplantation,the rough surface of regenerated tissue sunk obviously and the fibmblasts in all layer were found.While there were no regenerated tissue in the control side.Conclusion BMSCs seeded on HAAM could repair the articular cartilage defects of femoral intercondylar fossa from rabbits.
7.Effect of intraspinal grafting of adenovirus-mediated brain derived neurotrophic factor ex vivo transgene myoblasts cells and methylprednisolone on expression of caspase-3 after spinal cord injury
Qiang ZHANG ; Dewei ZOU ; Yong HAI ; Huasong MA ; Yongjun WANG
Chinese Journal of Trauma 2003;0(10):-
Objective To investigate the effect of intraspinal grafting of brain derived neurotrophic factor (BDNF) ex vivo transgene myoblasts cells and methylprednisolone on caspase-3 expression after spinal cord injury (SCI). Methods A total of 120 experimental rats were divided into Group A (spinal cord contusion injury group), Group B (grafting of BDNF ex vivo transgene myoblasts cells group), Group C (methylprednisolone intravenous injection group) and Group D (grafting of BDNF ex vivo transgene myoblasts cells and methylprednisolone intravenous injection group). At days 1, 3, 7, 14 and 28 respectively after SCI, the expression of caspase-3 was measured immunohistochemically for quantitative analysis via a computer image analysis system. The motion functional recovery of the rats was observed by praxiologic and electrophysiologic examination. Results Positive expression cells of caspase-3 were found in all groups, with number from the highest to the lowest in order of Group A, Group B, Group C and Group D (P
8.Curcumin inhibits the apoptosis ofHUVECs induced by oxidized low density lipoprotein
Dewei WANG ; Dongying ZHANG ; Li CAO ; Jian LIU
Basic & Clinical Medicine 2017;37(5):636-642
Objective To investigate the protective effect of curcumin on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL) and its possible mechanism.Methods Cultivated HUVECs were divided into six groups: control group, ox-LDL group, ox-LDL plus endoplasmic reticulum stress(ERS) inhibitor PBA group,curcumin group, ox-LDL plus curcumin group,ox-LDL plus curcumin plus PI3K inhibitor LY294002 group.Cell viabilities were evaluated by CCK-8 assays.The proportions of apoptotic cells were assessed by flow cytometry.The translocation of activating transcription factor 6 (ATF6) observed by laser confocal microscopy.Western blot was used to determine the expression of the ERS associated proteins:glucose-regulated protein 78(GRP78), protein kinase-like ER kinase(PERK), inositol-requiring kinase1(IRE-1) and the related pathways protein: LOX-1, AKT and phophorylated AKT.Results Compared with control group,increasedthe proportions of apoptotic cells(P<0.01),enhanced the expressions of ERS related proteins(P<0.01),promoted the transfer of ATF6 into the nucleus,as well as increased the expression of LOX-1(P<0.01)and decreased the expression of p-AKT(P<0.01) in the ox-LDL group;Compared with ox-LDL group,PBA inhibited ox-LDL-induced HUVECs apoptosis(P<0.01),curcumin inhibited ox-LDL-induced the expression of ERS associated protein and LOX-1(P<0.01), the nuclear translocation of ATF6, the apoptosis of HUVECs (P<0.01), and it also increased ox-LDL-induced down-regulation of p-AKT expression (P<0.01);LY294002 partially attenuated the inhibitory effect of curcumin on ERstress-related protein expression induced by ox-LDL(P<0.05).ConclusionsCurcumin can reduce ox-LDL induced apoptosis of HUVECs, its mechanism may be through the inhibition of LOX-1 expression and activation of AKT pathway to reduce ERS in cell.
9.The diagnostic and predictive value of long noncoding RNAs in ischemic heart disease
Li ZHANG ; Dewei WU ; Haiyan QIAN ; Mengyue YU
Chinese Journal of Laboratory Medicine 2015;38(12):877-880
Different types of noncoding RNAs (ncRNAs) are pervasively expressed in genomes of complex organisms.Current evidence suggests that long noncoding RNAs (lncRNAs) have significant roles at almost every stage of gene expression.It is an important regulatory factor involving in ischemic heart diseases,participating in the development of ischemic heart disease,expecting to predict early myocardial remodeling after myocardial infartion and heart failure,and attracting more and more attention.This review introduces the definition,classification,function and characteristics of lncRNA,and focuses on its diagnostic and predictive value in ischemic heart disease.
10.Homeostasis of complement system and the severity of coronary artery disease
Fei SONG ; Mengyue YU ; Jianru LIU ; Dewei WU ; Xinyue ZHANG
Chinese Journal of Laboratory Medicine 2016;39(9):685-689
Objective To investigate if the relative ratio between C1q and C3a, C5a had a relationship with the extent of coronary artery disease ( CAD) which had never been evaluated in humans.Methods Fifty-three patients scheduled for elective percutaneous coronary intervention ( PCI ) from February, 2016 to April, 2016 at Fuwai hospital were prospectively enrolled.According to the clinical and angiographic characters patients were divided into two groups:acute coronary syndrome ( ACS) group ( n=24), and control group (n=29, 19 patients with stable angina and 10 patients without CAD confirmed by angiography).In all individuals, fasting venous blood was collected by EDTA tubes after admission and strictly before PCI.The plasma level of C1q was measured by immune turbidimetric analysis, C3a and C5a were measured by ELISA tests.Differences between groups were assessed using t test, Mann-Whitney Utests, chi-squared test or Fisher exact test depending on the type of data respectively.Multivariate logistic regression analyses were conducted to evaluate the adjusted effect of C1q, C3a, C5a, C1q/C3a and C1q/C5a on ACS.Results Compared with control group, ACS group has an elevated circulation level of C3a (4 531.14 μg/L vs.4 179.95 μg/L, t=1.381,P=0.173) and C5a (6.44 μg/L vs.4.42 μg/L, t=0.133, P=0.108) but a decreased level of C1q (176.98 μg/ml vs.200.60 μg/ml, t=-2.022, P=0.048).The relative ratio of C1q/C3a was significantly decreased in ACS patients(4.05 ×10 -2 vs.4.97 × 10 -2 , t=-2.484, P=0.016).According to the multiple logistic regression analysis, lower relative ratio of C1q/C3a level proved to be independently associated with ACS ( OR=0.937, P=0.047, 95% CI:0.879-0.998).Conclusions The decreased relative ratio of C1q/C3a level proved to be independently associated with ACS.C1q/C3a ratio could be used as an important index reflecting the complement system homeostasis status which might have potential clinical value in evaluating the prognosis of patients with CAD.