2.Familial IgA nephropathy.
Chinese Journal of Pediatrics 2003;41(11):863-864
6.Extraction and identification of human adipose-derived stem cells
Wei WU ; Fang LIANG ; Xiaoqin SONG ; Pingan HU ; Min LIU
Chinese Journal of Tissue Engineering Research 2015;(28):4498-4502
BACKGROUND:Adipose-derived stem cel s are totipotent stem cel s in the adipose tissue, and have the function of self-renewal and multi-directional differentiation. Human adipose-derived stem cel s are ideal seed cel s with stable genetic milieu and few rejections.
OBJECTIVE:To extract human adipose-derived stem cel s from human omental adipose tissue and to identify the cel s by adipogenic and osteogenic induction.
METHODS:Omental adipose tissues were col ected from surgical patients to isolate and culture adipose-derived stem cel s using type I col agenase digestion, filtration and centrifugation. Cel growth was observed and proliferative curve of human adipose-derived stem cel s were drawn by cel counting method to calculate the doubling time at logarithmic growth phase. After adipogenic and osteogenic induction, induced cel s were identified using oil red O and alizarin red staining, respectively.
RESULTS AND CONCLUSION:Human adipose-derived stem cel s were successful y isolated from the omentum tissues of surgical patients. Adherent cel s were fusiform-shaped and like fibroblasts. The growth curve of passage 3 cel s was in S shape, and the doubling time was 45.90 hours. After adipogenic and osteogenic induction for 2 and 3 hours, respectively, oil red O staining showed unequal-sized orange fat droplets, and alizarin red staining showed typical calcified nodules that were in orange. These findings indicate that adipose-derived stem cel s have the adipogenic and osteogenic capacity.
7.Evaluation of the function of afferent neuronal pathways of the lower urinary tract in patients with spinal cord injury by using sympathetic skin response
Wei CHENG ; Hongliang LIU ; Bo SONG ; Min HOU
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(05):-
Objective To evaluate the function of afferent neuronal pathways (ANP) from the lower urinary tract (LUT) in patients with spinal cord injury (SCI) by use of sympathetic skin response (SSR). Methods Twenty-one patients with SCI (13 cases of incomplete injury, and 8 cases of complete injury) were recruited as a SIC group and 8 healthy volunteers as a control group. SSRs of all subjects were evoked by means of electrical stimulation (ES) of the median nerve and perineal region,as well as bladder filling (BF), while SSRs of the right palm and sole were recorded using surface electrodes. Results SSRs induced by ES of the median nerve and perineal region, and also by BF in the control group were recorded. SSRs of palms and soles could be recorded by using ES of the median nerve in patients with incomplete SCI, who had desire to void. However, SSRs could not be evoked in 3 of 13 patients with incomplete SCI but without sensation of perineal skin. In 8 patients with complete SCI but without sensation of trunk skin and bladder, SSRs of palms and soles could not be induced during ES of the median nerve if injuries were located over T_3, and SSRs of palms were recorded when the injuries were located between T_(4~9), while SSRs of palms and soles were evoked if injuries were located under T_(10). However, SSRs of palms and soles could not be evoked by ES of perineal region and BF in all patients. Conclusion SSRs, evoked by BF, could concord with the subjective sensation of the subjects from the LUT, and reflect the integrity of ANP from LUT. There is difference between somatosensory and viscerosensory ANP.
8.Isolation,purification and transplantation of rat islet cells
Sijiao CHEN ; Teimin LI ; Yangfeng LEI ; Min WEI ; Jindan SONG
Chinese Journal of Tissue Engineering Research 2007;11(12):2385-2388,2396
BACKGROUND: The islet cell transplantation has provided a solid basis for diabetic therapy, but the insufficient donor limits its development.OBJECTIVE: improving the method of isolating and purifying islets to observe the transplantation effect.DESIGN: A laboratory animal research.SETTING: Key Laboratory of Animal and Department of Cell Biology, China Medical University.MATERIALS: The experiment was carried out in the Key Laboratory of Animal and Department of Cell Biology in China Medical University between January and October in 2006. Donors were Wistar rats of either gender, weight 250-300 g;Acceptors were SD male rats, weight 180-220g. The two kinds of rats were all common closed population and from the Experimental Animal Department of China Medical University (The Admission Number of Experimental Animal Institute is SYXK(LIAO)2003-0013).METHODS: ①Isolation and exaltation of islet cells as well as the functional evaluation of pancreas: After etherisation, the Wistar rat without fasting was executed. A little cut was made on the beginning of the biliary pore, then the little cut lumbar anesthesia ductus, which were connected with a 1-mm-diameter syringe and full of cold collagenase solution (1.5 g/L), was inserted directly to dilate pancreas thoroughly. The pancreatic gland was isolated and digested in the water of centrifuge, when doing that, 1 mol/L NaOH was put interruptedly into the centrifuge tube to keep the pH value of the solution at 7.8±1.0. The rat pancreas purified by centrifugation of Ficoll density gradient: The identification of purified islets was evaluated by dithizone staining. The viability of islet was assessed by fluorescence staining of aridine orange and propidium iodide. The motility rate=the total number of live cells/(the total number of live cells + the total number of dead cells)×100%. Pancreatic activity was calculated: insulin release index=the level of insulin at the third hour (high concentration glucose)/the level of insulin at the second hour (low concentration glucose). ②The blood from vena caudalis of SD rat was sampled and measured the blood sugar after the intraperitoneal injection of streptozocin. The rat was diagnosed as DM when blood sugar was more than 16.7 mmol/L twice without fasting. The DM rats were divided into two groups, every group 8 rats. The experimental group rats were injected about 1 000 islet cells into the location below renal capsule, and the control group rats were injected the same volume of 1640 cultu re solution. Eight normal rats, whose glucose concentration ≤ 5.5 mmol/L, were taken randomly as normal controlled group. The blood sugar was measured every day after the surgery. The blood sugar less than 11.1 mmol/L without fasting was taken as the sign of successful islet transplantation. Intravenous sugar tolerance test was applied to the rats of normal control group, DM control group and experimental group 3 days after islet transplantation. Fasting for 12 hours before test, the blood sugar was measured at 0, 15, 30, 60, 90 and 120 minutes.MAIN OUTCOME MEASURES: Purity quotient, survival rate and activity of islet cells.RESULTS: All 24 SD acceptor rats were involved in the result analysis without miss.①The total number of purified islets of one pancreas was (1 150±141) in well morphology. The purity of islets was more than 95%. The viability of islets was more than 98%. ②The insulin secretion response to glucose challenge in vitro showed the mean value of insulin in the low-glucose medium was (70.5±6.9) mlU/L, while that of high-glucose medium was (321.4±11.6) mlU/L, the insulin release index was 4.6±0.52, that meant the beta cell of islet functioned well. ③The blood glucose level and the insulin level in plasma of the transplanted recipients restored to normal 3 days after transplantation. The survival period of transplanted islets was (6±2) days. But there was not any change in the concentration of blood sugar in the control group (16.7 mmol/L). The intravenous glucose tolerance test showed the identical outcome between the islet splantation group and the normal control group.CONCLUSION: There are high yield and high purify of islet cells in rats, which are isolated by in situ perfusion and purified by Ficoll density gradient centrifugation.
9.Induced differentiation of rat bone marrow mesenchymal stem cells into mesangial-like cells in vitro
Hong JING ; Min WEI ; Hongmei SONG ; Qiang QU
Basic & Clinical Medicine 2006;0(11):-
Objective To reconfirm the mesangial cell differentiation potential of BMSCs and to study the mechanism of cell induction.Methods Rat BMSCs were isolated and incubated with platelet-derived growth factor(PDGF)-BB to induce the mesangial cell phenotype.At day 7,the cell morphology,mesangial cell-specific markers and cell contraction function were examined.MAPK/ERK kinase inhibitor(PD98059) was added to examine its impact on cell growth and mesangial cells marker expression.Results After cultivation under the differentiation condition for 7 days,the induced cells expressed ?-actin and Desmin,which highly resembled cultured mesangial cells in rat.The induced cells with increasing level of PDGF receptors mRNA expression changed into a wide range of shapes from spindle to stellate and were contracted in response to angiotensin Ⅱ.PD98059 perturbed the induced cells mesangial cell-specific markers mRNA expression.Conclusion BMSCs can differentiate into mesangial-like cells with PDGF-BB induction in vitro.The mechanism involved in the differentiation was mediated through MAPK/ERK1/2 signal transduction pathway.