1.Application of tissue engineering in treatment of stress urinary incontinence
xiao-fei, SONG ; qiang, FU ; yue-min, XU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Stress urinary incontinence is one of the most common diseases in urinary system.At present,the major methods for treating stress urinary incontinence include medication,physico-behavior therapy and operation.However,for various reasons,the current methods do not yield satisfactory results.As a newly emerging technique,tissue engineering provides a new concept and method to treat stress urinary incontinence.The application of tissue engineering in the treatment of stress urinary incontinence is reviewed in this article.
2.Primary culture of adipose-derived stem cells and differentiation induction into myoblasts
xiao-fei, SONG ; qiang, FU ; yue-min, XU ; lei, CUI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(05):-
Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cells in induction group and control group,respectively.The induction lasted for 7 d,14 d,21 d,28 d and 35 d,respectively.Cell growth and cell morphology were observed by inverted phase contrast microscope,and immunofluorescence and flow cytometry were utilized to detect the expression of myoblast specific antigens desmin and myosin. Results ADSCs were successfully isolated and cultured,and were identified to be stem cells.On the 28th day of induction,cells in induction group displayed "swirl" morpholgy,and multinucleation was observed.It was revealed by immunofluorescence and flow cytometry that the highest expression rates of desmin and myosin were 52.57% and 50.04%,respectively on the 28th day of induction,while there was no expression before induction and in control group. Conclusion ADSCs can be isolated and cultured from rat adipose tissues,and can further differentiate into myoblasts after induction by culture media with 5-aza.The expression of myoblast specific antigen is the highest on the 28th day of induction.
3.Effects of siRNA interference combined with hyperbaric oxygen treatment on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage
Qiang PAN ; Lin ZHU ; Shuai ZHANG ; Puxian LI ; Xingtao DIAO ; Chunyu SONG ; Yong GAO ; Feng SI ; Qiang LI ; Xiao YUE
Chinese Journal of Geriatrics 2017;36(9):1010-1014
Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P<0.05)while,the combination therapy group achieved the best results(P< 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)%(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P<0.05) and the expression of Bcl-2 was higher(P<0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P<0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.
4.Intravascular biocompatibility of poly (3-hydroxybutyrate- co-3-hydroxyhexanoate)
Song WU ; Yinglong LIU ; Yue TANG ; Qiang WANG ; Feng WAN ; Xianghua QU ; Guoqiang CHEN
Chinese Journal of Tissue Engineering Research 2011;15(38):7066-7070
BACKGROUND: The degradable poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) has superior mechanical property and biocompatibility.OBJECTIVE: To elucidate the intravascular biocompatibility of PHBHHx in vivo.METHODS: We developed hybrid materials based on decellularized xenogenic vascular scaffolds that were coated with PHBHHx and implanted it into the abdominal aorta of New Zealand rabbits. The decellularized xenogenic pulmonary artery patch without PHBHHx coating served as the control. The implanted patches were determined for the histology, immunofluorescence staining, scanning electron microscopy and calcium contents at 1, 4 and 12 weeks after the surgery.RESULTS AND CONCLUSION: Hybrid patches exhibited smooth lumen surface without thrombus, the intimal hyperplasia was mild and recellularization was complete; immunofluorescence staining showed that the endothelial cells in the neointima were positive for CD31, with continuous single-layer arrangement, interstitial cells were positive for smooth muscle actin; the calcium content in hybrid patches was obviously lower than that in uncoated patches. PHBHHx shows a remarkable intravascular biocompatibility in vivo and is believed as an ideal candidate for lumen coating of cardiovascular tissue engineering.
5.Construction and characterization of cDNA library for IRM-2 mice
Qin WANG ; Jin LI ; Li SONG ; Qiang LIU ; Jingyin YUE ; Chuanjie MU ; Weisheng TANG ; Feiyue FAN
Chinese Journal of Radiological Medicine and Protection 2010;30(3):274-278
Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.
6.Correction of the epicanthal fold and angulus oculi using the Z-epicanthoplasty.
Ding-an LUO ; Yue-qiang ZHAO ; Song-shan WANG ; Hai-xiao ZHOU
Chinese Journal of Plastic Surgery 2007;23(6):491-492
OBJECTIVETo explore a new procedure for aesthetic correction of the medial epicanthal fold aim at the etiopathogenesis.
METHODSThe new Z-epicanthoplasty devise the upper and inferior margin of angle of eye medial as one angle of the Z.
RESULTSFrom 2004 to 2006, 129 patients were treated by using the method. Follow-up 6 to 24 months, all patients were satisfied by eliminating the medial epicanthal fold without obvious scar.
CONCLUSIONSThe method is more effect than traditionally Z-plasty. Our technique is a simple, advanced procedure that can be performed widely.
Adolescent ; Adult ; Blepharoplasty ; methods ; Eyelids ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Young Adult
7.Synergic effect of human IL-21 gene transfer combined with γ-ray irradiation on the growth of cervical carcinoma HeLa cells
Qin WANG ; Yingai WANG ; Li SONG ; Heng ZHANG ; Qiang LIU ; Yue FU ; Fenghua CHEN ; Feiyue FAN ; Jin LI
Chinese Journal of Radiological Medicine and Protection 2012;32(4):350-353
Objective To study the combined effect of interleukin-21 gene transfer and ionizing radiation on the growth of cervical carcinoma HeLa cells.Methods Previously constructed Ad-IL-21 gene was amplified by infecting 293A cells and the titer was measured by TCID50 method. HeLa cells were transfected with Ad-1L-21 and then irradiated with 6 Gy 137Cs γ-rays.The cells were divided into 5 groups,including blank control,Ad-LaeZ group,Ad-IL-21 group,radiation group and Ad-IL-21 combined with radiation group (combination group).The cell growth,cell cycle,apoptosis,and the expressions of IL-21 gene and protein in HeLa cells were detected.Results Ad-IL-21 was successfully amplified and the titer of Ad-11.-21 was 9 × 1010 pfu/ml.Compared with Ad-IL-21 group and radiation group,the cell growth of combination group was significantly inhibited at 96 h after transfection ( F =85.26,72.98,P < 0.05 ).The cells in combination group were arrested in G1 phase and decreased at S phase( F =36.69,34.83,P < 0.05),while the cellular apoptosis increased markedly ( F =28.23,25.57,P < O.05 ). The gene expression of 1L-21 in the combination group was 1.54- and 2.43-fold of Ad-IL-21 group and blank control group,respectively (F=22.31,36.65, P < 0.05 ), while the protein expression of IL-21 in the combination group was 1.62-fold and 2.31-fold of Ad-IL-21 group and blank control group,respectively ( F =27.36,35.86,P < 0.05 ).Conclusions Ad-IL-21 gene transfection combined with radiation has synergic effect on the inhibition of cervical carcinoma cell growth.
8.Investigation of acupuncture in improving sleep, cognitive and emotion based on attenuation of oxidative stress in prefrontal cortex in sleep-deprived rats
Fei-Yi ZHAO ; Sheng-Nan GUO ; Yan XU ; Hong XU ; Guo-Hua WANG ; Hua-Ling SONG ; Li-Ping YUE ; Fang-Lei CHEN ; Si-Han CHEN ; Qiang-Qiang FU
Journal of Acupuncture and Tuina Science 2021;19(3):157-166
Objective: To explore whether acupuncture can improve sleep disturbance, cognitive impairment and emotional disorders caused by sleep deprivation, and its association with the attenuation of oxidative stress injury in prefrontal cortex. Methods: Fifty-two male Sprague-Dawley rats were randomly divided into a control group (n=10), a model group (n=14), a manual acupuncture (MA) group (n=14), and a sham-MA group (n=14). All the groups were established as sleep deprivation models via the modified multiple platform method, except for the control group. Rats in both the MA group and the sham-MA group received corresponding intervention, respectively. After modeling and intervention, the four groups received three behavioral tests, namely sleep monitoring, by comprehensive lab animal monitoring system (CLAMS), Morris water maze (MWM) test and open-field test (OFT), followed by oxygen free radical level test and Western blot (WB) detection for the expression levels of Bax and Bcl-2. Results: The MA group derived more sleep time within 24 h than either the model group or the sham-MA group (both P<0.05). On MWM orientation navigation test day 1, there were no significant differences in escape latency among the control, MA and sham-MA groups (P>0.05), and the escape latency was significantly shorter in these three groups than that in the model group (all P<0.05). On test day 4, the escape latency was markedly shorter in the MA group than that in either the model group or the sham-MA group (both P<0.05); meanwhile, the MA group showed significantly better performance compared with these two groups in space probe test (both P<0.05). In OFT, compared with the control group, there was a significant decline in the horizontal movement score in the other three groups (all P<0.05), and the decrease was more significant in the model group and the sham-MA group than that in the MA group (both P<0.05). The superoxide dismutase (SOD) content was markedly higher and the malondialdehyde (MDA) content was markedly lower in the MA group than those in the model group and the sham-MA group (all P<0.05). Compared with the model group and the sham-MA group, the expression of Bax was significantly lower and the expression of Bcl-2 was significantly higher in the MA group (all P<0.05). Conclusion: MA therapy can lengthen the sleep time in sleep-deprived rats and improve learning and memory impairments induced by sleep deprivation, and the underlying mechanism may be associated with the enhancement of antioxidant capacity in the prefrontal cortex and the inhibition of hippocampal neuronal apoptosis.
10.Effect of hyperbaric oxygen on the scar formation at the rabbit ears at an early stage.
Qiang ZHANG ; Jia-Song SHAO ; Yi-Gang YUE ; Hai ZHOU ; Ming-Chin HUA ; Min ZHANG
Chinese Journal of Plastic Surgery 2013;29(1):55-58
OBJECTIVETo observe the effect of hyperbaric oxygen on the scar formation at the rabbit ears at an early stage.
METHODS16 New Zealand rabbits were used to establish the hypertrophic scar model on the ears, 4 wounds on each ear. The rabbits were randomly divided into hyperbaric group(n = 8) and control group (n = 8). The rabbits in the hyperbaric group received hyperbaric oxygen treatment, inhaling oxygen for 1 hour daily under 2ATA circumstance until the wounds were healed. The wound healing and the scar size, thickness, color and hardness in the ears were recorded. After healing, the scar was taken for histologic study with HE staining, Masson staining and trinitrophenol sirius red staining.
RESULTSIn hyperbaric oxygen group, the healing time of 64 wounds was (16.7 +/- 1.8) d, while it was (20.2 +/- 2.3) d in the control group, suggesting a significant difference between two groups (P < 0.05). The incidence of hyperplastic scar was lower (38/64, 59. 4% ) in hyperbaric oxygen group than that (52/ 64, 81.2%) in control group with significant difference between them (P < 0.05). Compared with the control group, the hyperbaric oxygen group had thicker corium layer and less fibroblasts, well-arranged but rare collagen and less collagen nodus and vortex-like structure under microscope. The hyperplastic index of scar was 3.48 +/- 0.94 in hyperbaric oxygen group and 4.65 +/- 0.76 in control group respectively (P < 0.01). The density of fibroblast in two groups were 186.5 +/- 27.3 (hyperbaric oxygen group) and 246 +/- 41.6 (control group) with statistically significant difference (P < 0.05). Furthermore, the area density of collagen fiber were (31.42 +/- 5.36)% in hyperbaric group and (43.62 +/- 7.36)% in control group (P < 0.05). The amount of collagen I and III was (71.42 +/- 5.36)% and (28.58 +/- 5.36)% in hyperbaric oxygen group, (62.46 +/- 7.32)% and (37.54 +/- 7.32)% in control group (P < 0.05). The ratio of collagen I to III was 2.499 in hyperbaric oxygen group, which was similar to the ratio (4:1) in normal skin, compared with the control group (1.664).
CONCLUSIONSThe hyperbaric oxygen can promote wound healing and effectively inhibit the early hyperplastic scar in rabbits ears.
Animals ; Cicatrix ; Disease Models, Animal ; Ear ; pathology ; Female ; Hyperbaric Oxygenation ; Male ; Rabbits ; Wound Healing