1.Effects of systemic irradiation and W11-a12 on neutrophils in wounds
Xiaohong CHEN ; Tianmin CHENG ; Guoping AI
Journal of Third Military Medical University 2001;23(3):287-289
Objective To study the effects of systemic irradiation and conglutinant drug W11-a12 on the number and some functions of wound nentrophils (Neu). Methods Wound Neu was collected from sponges which were implanted in rat's dorsum incision. The number of Neu, as well as the phagocytic function and motility of wound Neu were measured. Results After 4,6,8 Gy systemic irradiation, the number of white blood cells and Neu in wound, as well as the phagocytic function and chemotactic motility of wound Neu, were significantly decreased at 24 h, 48 h after wounding. W11-a12 markedly increased the number of wound Neu, improved the phagocytic function and chemotactic motility of wound Neu at 24 h, 48 h after wounding despite the rats were radiated or not. Conclusion The results indicated that the decreased number and function of wound Neu in the early stage of wound healing contributed to the impairment of repair after systemic irradiation. W11-a12 accelerated normal and irradiation-impaired wound healing partly by increasing the number of wound Neu and improving the Neu function.
2.Effect of a novel isoflavone compound on plasma lipid and cholesterol of ovarectomied rats
Xiuming CHEN ; Guoping AI ; Yongping SU ; Lilong ZHANG ; Qing CHANG
Journal of Third Military Medical University 2003;0(16):-
Objective To investigate the effect of a novel isoflavone compound(F11) on the plasma lipid and cholesterol of the ovarectomied rat.Methods Female SD rats at age of 3 months old were randomly divided into 6 groups,that is,sham operation group(Sham),normal saline group(2 ml/d),estradiol group(E2,50 ?g?kg-1?d-1),and 3 F11 groups(15,50,150 mg?kg-1?d-1).Besides the Sham group,the ovary of the rats from other groups were resected,and received the injection as above mentioned.All rats were killed 10 weeks later,and their plasma lipid,total cholesterol,LDL,HDL,and body weight and uterine weight were measured.Results The plasma lipid,total cholesterol,LDL,HDL were significantly different in normal saline group and 4 treatment group(P
3.Biological effect of four isoflavone derivatives on proliferation of endometrial epithelial cells in vitro
Cheng CHEN ; Qing CHANG ; Lilong ZHANG ; Guoping AI
Journal of Third Military Medical University 2003;0(16):-
Objective To study the biological effects of four isoflavone derivatives(F8,F11,ZF3 and ZF7)on endometrial epithelial cells.Methods The endometrial epithelial cells were cultured through collagenase enzymatic digestion and twice grit filtration.The biological effects on endometrial epithelial cells of four isoflavone derivatives were compared through MTT.Results The primary endometrial epithelial cells were successfully disassociated,cultured and passaged down stably.Cell proliferation was significantly increased by F8(25 mol/L)(P
4.Platelet-derived Growth Factors and Tissue Repair
Guohe YAN ; Guoping AI ; Zhongmin ZOU ; Yongping LI
Chinese Journal of Rehabilitation Theory and Practice 2007;13(1):39-41
Platelet-derived growth factors (PDGFs) are a member of family of glycoprotein dimers (Mr 27000~35000) that exert potent mitogenic and chemotactic activities toward cells of mesenchymal origin. The PDGFs possess a myriad of critical roles in embryonic development,cellular differentiation and response to tissue damage. It is one of the growth factors,which appear early during the process of wound healing. Especially,PDGFs can effectively promote the healing of some chronic refractory wounds,such as diabetes mellitus ulcer,chronic venous ulcer,bedsore,radioactivity ulcer,etc..
5.Cultivation and isolation of the bone marrow mesenchymal stem cells
Guoping AI ; Yongping SU ; Guohe YAN ; Xingze RAN ; Xiaohong LIU ; Chengji LUO ; Tianmin CHENG
Journal of Third Military Medical University 2001;23(5):553-555
Objective To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.
6.Effect of long-term depleted uranium ingestion on testosterone production and the expression of StAR and P450sec mRNAs in rats
Rong LI ; Yanbing LENG ; Jiong REN ; Jing LIU ; Guoping AI ; Hui XU ; Yongping SU ; Tianmin CHENG
Chinese Journal of Radiological Medicine and Protection 2009;29(1):13-16
Objective To explore the effect of long-term depleted uranium (DU)ingestion on testosterone production in rats, and its involvement mechanism. Methods Male and female rats (F0 and F1 respectively) for 160 days, respectively. The contents of testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) in serum were detected in 20 months of F0 generations, and 15 months of F1 generations. RT-PCR was used to analyze the levels of StAR mRNA and P450scc mRNA. Results Compared with the normal control group, the testosterone contents in exposed F0 and F1 generations increased, the lowest was 51.73 U/L, but those of LH and FSH decreased. The expression of StAR mRNA in the low-doze group of F1 generation (StAR/β-actin = 1.35) was up-regulated, down-regulated for other groups.compared with the normal control group (P450scc/β-actin = 0. 313), the expression of P450scc mRNA in the low- and high-dose groups of F0 generation were decreased (P450scc/β-actin = 0.21), and those in the low- and high-dose groups of F1generation were increased (P450scc/β-actin = 0.623) (P ≤ 0.01). Conclusion Long-term DU exposure inhibit the male reproduction by intervening the sexual hormone production through down-regulated the expression of StAR mRNA and P450scc roRNA.
7.Construction of eukaryotic expressing vector pEGFP-N1/PDGF-A for transducting Dermis-derived mesenchymal stem cells
Guohe YAN ; Yongping SU ; Junping WANG ; Daijie WANG ; Guoping AI ; Fengchao WANG ; Xinze RAN ; Tianmin CHENG
Journal of Third Military Medical University 2003;0(20):-
Objective To clone platelet-derived growth factor A chain (PDGF-A) gene and insert PDGF-A gene into. Enhanced green fluorescent protein (EGFP) vector and then transformed into dermis-drived mesenchymal stem cells (DMSCs). Methods cDNA clones encoding human PDGF-A gene were isolated from a human hepatoma cell line mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The PCR amplified fragment of PDGF-A gene was cloned into pMD18-T vector. The eukaryotic expression vector pEGFP-N1/PDGF-A was constructed by subcolone PDGF-A gene into pEGFP-N1 vector. PDGF-A gene was transfected into DMSCs with the help of Fugene 6 transfection reagent. Results Full cDNA sequence encoding human PDGF-A gene had been cloned, which sequence was consistent with the reported sequence in GenBank by sequence assaying. Conclusion cDNA sequence encoding human PDGF-A gene was successfully cloned into pEGFP-N1. The transient expression of PDGF-A gene in DMSCs has been realized.
8.The advances in the application of amniotic membrane stroma in promoting tissue repair.
Guohe YAN ; Guoping AI ; Yongping SU
Journal of Biomedical Engineering 2002;19(4):676-679
Amniotic membrane is composed of amniotic epithelium, basement-membrane and stroma. Amniotic membrane is an easily obtained biomaterial and easily to be also processed, preserved and transported. However, its applicability will not be destroyed after it has been preserved for a long time(about one year). Thus it has been utilized widely in laboratory and clinical surgery. Generally, homologous amniotic membrane does not induce rejection after allotransplantation, and it is a bio-absorbable and degradable material. The purpose of this paper is to review the characteristics of amniotic membrane that makes it potentially useful in promoting tissue repair.
Amnion
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transplantation
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Biological Dressings
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Humans
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Tissue Engineering
9.Human amniotic membrane loaded with porcine keratinocytes for constructing epidermal substitute of skin
Guohe YAN ; Yongping SU ; Feng WANG ; Guoping AI ; Tianmin CHENG ; Huaien ZHENG ; Xinze RAN ; Hong XIAO ; Chongfu TAN
Chinese Journal of Tissue Engineering Research 2005;9(22):245-247
BACKGROUND: As a kind of semitransparent membrane, human amniotic membrance contains many kinds of nutrients, which is a good biological material loaded with keratinocytes.OBJECTIVE: To construct epidermal substitute of the skin from human amniotic membrane loaded with porcine keratinocytes and examine the morphological characteristics of the growth and proliferation of keratinocytes seeded on human amniotic membrane.DESIGN: Single sample study and repetitive measured observation based on the cells.SETTING: Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed in the State Key Laboratory of Trauma, Burn and Combined Injury and Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA from January to November 2001. Porcine keratinocytes was collected from Guizhou minipigs aged 3 weeks.METHODS: The primarily cultured keratinocytes of Guizhou minipigs were subcuhured, expanded and bred on the stroma surface of human amniotic membrance at the density of 1.63 × 105/cm2. The growth and proliferation of keratinocytes were observed under inverted microscope every day. From the 3rd day and the 15th day after being cultured, the growth of keratinocytes on human amniotic membrane was examined under light microscope and electron microscope.MAIN OUTCOME MEASURES: The growth of keratinocytes on human amniotic membrane was examined RESULTS: Keratinocytes evidently adhered to the stroma surface of human amniotic membrane about 30 minutes after being cultured, which was observed under inverted microscope. Most keratinocytes grew and adhered to the stroma surface of human amniotic membrane within 24 hours. Monolayer of keratinocytes formed and completely covered human amniotic membrane within 3 days. It was observed under the light microscope that the monolayer of keratinocytes adhered to human amniotic membrane and arrayed tightly. The keratinocytes presented in the shape of polygon, and plasmalemmas of keratinocytes formed many pseudopods under the observation with scanning electron microscope. Keratinocytes adhered to human amniotic membrane well and with many keratinofilaments in them under the observation with transmission electron microscope. Keratinocytes arrayed on human amniotic membrane densely with many cellular debris and some keratinocytes formed cavitations in them due to aging after growth for 15 days under the observation with inverted microscope.CONCLUSION: Human amitotic membrane is a good carrier of keratinocytes cultured on it in vitro, and is able to promote the proliferation of keratinocytes significantly. However, when keratinocytes were loaded on the human amniotic membrane for 15 days, some keratinocytes formed cavitations in them due to aging.
10.CXCR4 gene transfection enhancing the distribution of dMSCs to the wounded skin of rats with combined wound and irradiation injury
Zhaowen ZONG ; Tianmin CHENG ; Xinze RAN ; Yongping SU ; Shiwu DONG ; Nan LI ; Junping WANG ; Guoping AI ; Zheng LI
Chinese Journal of Radiological Medicine and Protection 2009;29(4):351-354
Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.