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3.
Article in Chinese | WPRIM | ID: wpr-424128

ABSTRACT

Objective To observe whether dermal multipotent stem cells (dMSCs) treated with platelet-derived growth factor-AA ( PDGF-AA )could distribute more frequently to the bone marrow in rats of total body irradiation (TBI).Methods Male dMSCs were isolated and 10 μg/L PDGF-AA was added to the culture medium and further cultured for 2 h.Then the expression of tenascin-C were examined by Western blot, and the migration ability of dMSCs was assessed in transwell chamber.The pre-treated dMSCs were transplanted by tail vein injection into female rats administered with total body irradiation, and 2 weeks after transplantation, real-time PCR was employed to measure the amount of dMSCs in bone marrow.Non-treated dMSCs served as control.Results PDGF-AA treatment increased the expression of tenascin-C in dMSCs, made (1.79 ± 0.13) × 105 cells migrate to the lower chamber under the effect of bone marrow extract, and distributed to bone marrow in TBI rats, significantly more than ( 1.24 ± 0.09) ×105 in non-treated dMSCs (t = 8.833, P < 0.0l ).Conclusions PDGF-AA treatment could enhance the migration ability of dMSCs and increase the amount of dMSCs in bone marrow of TBI rats after transplantation.

4.
Article in Chinese | WPRIM | ID: wpr-396257

ABSTRACT

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.

5.
Article in Chinese | WPRIM | ID: wpr-393494

ABSTRACT

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.

6.
Chinese Journal of Biotechnology ; (12): 291-296, 2008.
Article in Chinese | WPRIM | ID: wpr-276125

ABSTRACT

DNA fragment containing human alpha-defensin 5 mature peptide (mHD-5) coding sequence with biased codons of E. coli was amplified by PCR, which was subsequently cloned into the plasmid pMAL-p2x in order to create pMAL-p2x-mHD-5 expression vector. The plasmid pMAL-p2x-mHD-5 was transferred into engineered strain BL21(DE3) to express heterogeneous fusion protein (MBP-mHD-5). The soluble MBP-mHD-5 targeted protein inducible expressed by IPTG was accounted for about 30% under optimized conditions. The recombinant mHD-5 (rmHD-5) peptide was successfully purified through a separation process including affinity chromatography, Factor Xa digestion and ion exchange chromatography. The bioactivity of rmHD-5 was examined by bacteria-inhibition tests in liquid culture. The growth of E. coli ATCC25922 was dramatically suppressed with an inhibition rate of 90%, with the presence of 62.5 microg/mL rmHD-5 in the media. These results indicate that the strategy of soluble expression of fusion protein in E. coli can be a useful and practical way to produce bioactive defensins.


Subject(s)
Humans , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Recombinant Fusion Proteins , Genetics , Solubility , Transformation, Bacterial , alpha-Defensins , Genetics
7.
Article in Chinese | WPRIM | ID: wpr-407853

ABSTRACT

BACKGROUND: Human amniotic membrane (HAM) contains various ingredents such as collagen, glycoprotein,proteoglycan, integrin and laminated body, and so on, and expresses many kinds of growth factors and mRNA-associated proteins. And these ingredents can supply abundant nutriments for cellular proliferation and differentiation, and benefit cells to grow and propagate. Whether or not HAM can load porcine bone marrow-derived mesenchymal stem cells (BMSCs) to well grow on it deserves to be further investigated.OBJECTIVE: To set up a method of tissue engineering of human amniotic membrane loading porcine BMSCs and observe the morphological characteristics of growth and proliferation of BMSCs seeded on HAM.DESIGN: Randomized controlled observation.SETTING: State Key Laboratory of Trauma, Burn and Combined Injury, General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out in the State Key Laboratory of Trauma, Burn and Combined Injury,General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA between January and November 2003. Three Guizhou minipigs of either gender, aged 2 to 3 months, weighing from 6 to 8 kg, were provided by the Experimental Animal Center, Third Military Medical University of Chinese PLA. Main reagent:ISCOVE'S modified DULBECCO'S medium (IMDM) culture medium (Hyclone, USA); high-quality fetal bovine serum PAA (Germany); haematoxylin (China); Eosin B (Sigma, USA) and OCT embedding medium (USA). Main instruments: BX51 stereoscopic fluorescence microscope (Olympus, JaPan); IX70 inverted fluorescence microscope (Olympus, Japan);cryostat (2700-Frigcut, Germany); myeloid puncture needle (Jiangsu); superclean bench (Sujing Bloc Antai Company);CO2 constant-temperature incubator (QUEUE, USA).METHODS: HAM was prepared as previously described. The BMSCs of Guizhou minipigs isolated and cultured according to method described previously were primarily cultured and passaged, then they were inoculated to the stromal surface of HAM at different densities (0.84×105 cells/cm2,1.54×105 cells/cm2,2.75×105 cells/cm2); The growth and proliferation of BMSCs of different densities were observed under an inverted microscope and scanning electron microscope; BMSCs of the second or the third passages were inoculated on HAM held with tissue-holding device at a density of 1.54×105 cells/cm2, and they were cultured for 18 days at most. The HAM was daily rolled, sliced and stained by HE for observing the growth of BMSCs loaded on HAM under the light, scanning and transmission electron microscopes.MAIN OUTCOME MEASURES: The growth of BMSCs on HAM was examined at different densities and different time points.RESULTS: ① Comparison of growths of BMSCs promoted by different densities of HAM: BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 were irregular and scattered under an invert microscope. Distances between BMSCs were biggish. BMSCs seeded on HAM at the density of 1.54×105 cells/cm2 were regular in arrangement and moderate in density, with clear cell outline and good cell activity before 24 hours, and seeded at the density of 2.75×105 cells/cm2 were congested with many nonattached cells and the longer the growing time of the cells was, the more the cellular debris were observed. BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 under the scanning electron microscope, scatted on HAM presented in shapes of irregular, long, thin and flat polygon. Their membrane protuberances presented in shapes of thick and thin, and the distances between cells were biggish. BMSCs,which were planted on HAM at the density of 1.54×105 cells/cm2 have similar appearance of their bodies and membrane protuberances, and the membrane protuberances were more compared with the BMSCs planted at the density of 0.84×105 cells/cm2. Their membrane protuberances intercrossed each other, and the margin of some BMSCs overlapped each other. BMSCs planted at the density of 2.75×105 cells/cm2, arraved on HAM crowdedly and overlappedly with many debris. Their membrane protuberances were not obviously. The margin of some BMSCs was overlapped.② Comparisonof growths of BMSCs promoted by HAM at different time points: Under the inverted microscope, the BMSCs adhered quickly to HAM after being incubated for about 30 minutes. All of BMSCs adhered to HAM within 24 hours, and formed monolayer on it within 48 hours, and grew densely on HAM after being cultured for 4 to18 days. Under the light and electron microscopes, HE results revealed that BMSCs adhered tightly and grew on HAM in different arrays, such as emitting, whirlpool or parallel,and their nuclei located in middle, dense in staining, were big and clear. The shapes of BMSCs were comparatively consistent on HAM. HAM loaded with BMSCs grew 4 days, and BMSCs covered HAM completely. The densities of BMSCs on HAM were suitable, and their bodies were large, and presented irregular, long,thin and flat polygon under the scanning electron microscope. The margin of some BMSCs overlapped each other. The protuberances of cellular membrane of BMSCs were abundant in the shapes of thick and thin. Some protuberances intercrossed each other in the shape of net. BMSCs adhered tightly to HAM through these protuberances. HAM loading BMSCs grow 4 days; most of BMSCs grew on HAM in double layers with the shapes of cambiform under the transmission electron microscope, Their nucleoli were clear. The protuberances of cellular membrane of BMSCs, which situated at two sides of nuclei and overlapped each other, were long. Most of chromatins of BMSCs were autosome.Abundant organell such as rough endoplasmic reticulum (RER),mitochondria could be observed in BMSCs.CONCLUSION:HAM is able to promote the proliferation of BMSCs significantly. BMSCs may be cultured on HAM ex vivo.HAM is a good carrier of BMSCs.

8.
Article in Chinese | WPRIM | ID: wpr-408220

ABSTRACT

BACKGROUND: Cervical sympathetic ganglia block accelerates the re covery of the homeostasis of organic nervous-endocrine-immune system, butit is still unclear whether it can suppress the imbalance of homeostasis in duced by post-traumatic stress disorder. OBJECTIVE: To observe the effect of cervical sympathetic ganglia blockon the mortality of mice with combined radiation and burn injury, andwhether it can become an easy and effective method to treat secondarydamage after serious trauma. DESIGN: A randomized grouping design, an animal controlled experiment. SETTING: Department of Anesthesiology, Guangzhou General Hospital, Guangzhou Military Area Command of Chinese PLA.MATERIALS: The experiments were carried out in the Institute of Combined Injury, the Third Military Medical University of Chinese PLA between February 2004 and July 2005. Totally 160 Kunming mice were randomly divided into control group (n=50) and cervical sympathetic ganglia block group (n=50). In the control group, the mice were only induced to models of combined radiation and bum injury, and treated with injection of 0.3 mL saline at cervical part. In the cervical sympathetic ganglia block group, the mice were induced to models of combined radiation and burn injury, and then treated with cervical sympathetic ganglia block, once a day for 14 days continuously.METHODS: Methods to induce injury in the animals: ① Radiation injury: The mice were given even radiation of 60Coγ ray (5 Gy) at a distance of 1.5 m to the whole body, the rate of absorptive dosage was (5.17-5.33) mGy/s. ② Burn injury: After the radiation injury, coagulated gasoline was smeared on the back and burnt for 8 s to induce degree Ⅲ burn injury of 15% of the total body surface, which was proved by the pathological section. Methods of cervical sympathetic ganglia block: Cervical sympathetic ganglia block was given bilaterally, and then the mice were injected with 0.2 mL lidocaine (5 g/L), and it was observed whether the symptoms similar to Horner syndrome (hyperemia of conjunctiva, drooping eyelid,blushing, smaller eyeslit) occurred or not at 5 minutes after injection.MAIN OUTCOME MEASURES: The mortality at 2, 5, 7, 10, 20 and30 days after injury and the changes of the numbers of red blood cells,white blood cells and blood platelet in peripheral blood at 7, 14 and 21 days after injury were observed in both groups. The effects of cervical sympathetic ganglia block on the levels of tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum at 3, 6 and 14days after combined radiation and burn injury were also observed.RESULTS: All the 160 mice were involved in the analysis of results without deletion. ① Compared with the control group, the mortalities at 5,7, 10, 15, 20 and 30 days in the cervical sympathetic ganglia block group were significantly decreased [control group: 8%, 22%, 32%, 54%, 74%,82%, 90%; cervical sympathetic ganglia block group: 8%, 14%, 16%, 22%,28%, 34%, 56%]. ② Compared with the control group, the numbers of red blood cells, white blood cells and blood platelets in peripheral blood at 7,14 and 21 days after injury in the cervical sympathetic ganglia block group were significantly increased [at 21 days: red blood cells: 23.21×1012 L-1, 14.58×1012 L-1; blood platelet: 16.87×1011 L-1, 12.57×1011 L-1; white blood cells: 20.65×109 L-1, 14.58×109 L-1]. ③ The levels of TNF-α, IL-1β andIL-6 in serum at 3, 6 and 14 days after injury in the cervical sympathetic ganglia block group were significantly decreased as compared with those in the control group [at 14 days: TNF-α: 189, 365 ng/L; IL-1β: 14, 23 ng/L;IL-6: 70, 132 ng/L].CONCLUSION: Cervical sympathetic ganglia block can significantly decrease the mortality of animals with combined radiation and burn injury,and it is an easy and effective method to treat serious trauma, and the mechanism may be realized through accelerating the recovery of hematopoietic function and suppressing the excessive inflammatory reaction.

9.
Article in Chinese | WPRIM | ID: wpr-409306

ABSTRACT

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.
Article in Chinese | WPRIM | ID: wpr-560354

ABSTRACT

Objective To investigate the effect of half-body ionizing radiation on the concentration of MDA, SOD in serum and the spleen index, CFU-S and the protective function of supplementary taurine (Tau) in half-body ionizing irradiated mice. Methods Kunming mice were randomly divided into 6 groups: normal control, total-body irradiation (TBI), total-body irradiation+Tau, left-half-body irradiation, half-body irradiation+Tau, and total-body shielding irradiation. The relevant indexes such as the spleen index, spleen colony forming units (CFU-S) were observed. The ionizing radiation was performed under ~ 60 Co ? ray with absorbed dose 8.0 Gy, dose rate 68.46 cGy/min. Taurine at dose of 500 mg/kg was injected intraperitoneally twice a day before irradiation, once 30 min before irradiation, and once 6 h after irradiation. Results In TBI group, the spleen index and CFU-S were decreased remarkably, MDA increased and SOD reduced in serum. The hematopoietic function of spleen was not improved by supplementary taurine (P

11.
Article in Chinese | WPRIM | ID: wpr-563879

ABSTRACT

Objective To measure the collagen content in the granulation tissue of skin wound surface in rats with depression in order to explore the possible mechanism about depression to wound healing.Methods Totally 126 Sprague-Dawley rats were randomly and equally allocated to 3 groups: simple wound group,depression plus wound group,wound plus depression group.Unpredictable stimuli,including fasting,electrocution,water deprivation,swimming in cold water and others,were performed for 18 d to establish depression model of rats.A full-thickness dermal wound(2.2 cm in diameter) was made on the back of rats to build wound model.Rats from depression plus wound group received 18 days' stimuli and then wound cutting.For those of wound plus depression,stimuli were carried out from the next day of wounding.The basal tissue in the center of the wound was taken out from 6 rats of every group in 3,5,7,10,14,18 and 21 d after wounding.Hydroxyproline reagent kit was used to detect the content of hydroxyproline in collected tissue.Wound collagen depositions in every time point were stained by nitroxanthic acid sirius red and then observed by polarization microscopy.Results Hydroxyproline content and collagen deposition in wound granulation tissue of depression plus wound group rats were lower than those in the other 2 groups.From 18 d after wounding,rats of depression plus wound group had a tendency to the same level as wound plus depression group.Conclusion Depression plus wound significantly decreases hydroxyproline synthesis,reduces collagen deposition in wound,and then delays the wound healing.

12.
Article in Chinese | WPRIM | ID: wpr-679124

ABSTRACT

Objective To evaluate the biological effect of a kind of protective cover against inhalation of depleted uranium (DU) dust particles. Methods At 1 and 3 d after inhalation of DU dust particles, uranium concentrations in the blood, lung, bronchia, kidney, and liver of the rats in the protected group and non protected group were measured and the efficiency of protection was calculated. Results Uranium levels in all tissues and fluids of the rats in the protected group decreased significantly to 71.2%-96.1% as compared with those in non protected group. Conclusion The protective cover is effective to reduce the harm due to inhalation of DU dust particles.

13.
Article in Chinese | WPRIM | ID: wpr-678850

ABSTRACT

Objective To find out the best method for elimination of uranium contamination in rat tissues as low as possible by removal of shrapnel fragments. Methods Experimental rats were divided into six groups: route group, decontamination before surgery group, decontamination in incision group, changing surgical appliances group, removing tissues around group, and comprehensive method group. Uranium concentrations in tissues and fluids in all groups were measured at 7, 14, and 21 d after operation. The efficiency of decontamination by different methods was compared. Results The highest uranium concentration in tissues was found in the route group, but the lowest in the comprehensive method group, and the second lowest in removing tissues around group. Conclusion The comprehensive method is the best one in all of the surgical removal methods. The soft tissues around DU shrapnels should be removed if they are not critical organs.

14.
Chinese Medical Journal ; (24): 878-882, 2003.
Article in English | WPRIM | ID: wpr-294211

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of dosages of total body irradiation on the healing process of cutaneous wounds and to observe the changes of wound area at different periods after injury.</p><p><b>METHODS</b>The entire body irradiation from a (60)Co gamma-ray source was performed on Wistar rats. The single dosage varied from 1 to 8 Gy. Within 1 h after irradiation, two whole thickness circular cutaneous wounds corresponding to 2.5% of total body surface area (Phi = 22 mm) were produced on the back of the animals (combined injury groups). Same wounds were produced on rats with no irradiation (single wound group). Wound healing was observed at different points after injury.</p><p><b>RESULTS</b>After total body irradiation with the dose of 1, 2, 3, 4, 5, 6, 7 or 8 Gy, the wound healing was obviously retarded as the dosages increased. The wound area remained was larger in the large dosage groups than in the small dosage groups. Seven days after injury, there was 33.5% wound surface left unhealed in the single wound group, whereas in the combined injury groups, 35.4%, 38.1%, 41.6%, 48.8%, 53.9%, 63.7%, 69.2% and 73.9% of the wound surfaces remained unhealed, respectively. Statistical analysis showed marked correlations between the various times after total body irradiation and various dosages to the percentage of unhealed wound surface. Nine dose-effect relation formulae were deduced according to the statistical results.</p><p><b>CONCLUSIONS</b>In soft tissue trauma combined with radiation injury, the delay of wound healing is related to the dose of radiation inflicted. It is also related to the time between injury and time of observation.</p>


Subject(s)
Animals , Female , Male , Rats , Dose-Response Relationship, Radiation , Rats, Wistar , Time Factors , Whole-Body Irradiation , Wound Healing , Radiation Effects
15.
Article in Chinese | WPRIM | ID: wpr-557468

ABSTRACT

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.

16.
Article in Chinese | WPRIM | ID: wpr-677975

ABSTRACT

Objective To construct a bait vector containing human glucocorticoid receptor(GR) ligand binding domain (LBD) in yeast two hybrid system in order to screen its cDNA library Methods PCR was used to amplify GR LBD fragment from the fetal liver cDNA library with the primers designed in accordance with the sequence in GenBank GR LBD The product was inserted into pGEM(r) T Vector Systems After verified with restriction endonuclease digestion of SmaⅠ/SalⅡ, the vector was inserted into the "bait plasmid" pGBKT7 (named as pGBKT7 GR LBD) After confirmation with restricted endonuclease digestion and sequence analysis, the plasmid was transformed into the yeast cell AH109, and the transformants were selected on SD/ Trp plates The interaction between GR and SRC 1 was tested by ? gal activity with yeast two hybrid system Results The amplified product of 893 bp was inserted into pGEM(r) T Vector and proven to be successful with double restriction enzyme digestion Sequence analysis revealed that the sequence was correctly inserted into pGBKT7 with a right reading frame AH109 [pGBKT7 GR LBD] grew on SD/ Trp plates, but not on the other selective media GR could interact with SRC 1 Conclusion The bait plasmid pGBKT7 GR LBD constructed expresses GR LBD correctly, and can't activate the transcription of reporter gene alone in yeast two hybrid system

17.
Chinese Medical Journal ; (24): 1763-1766, 2002.
Article in English | WPRIM | ID: wpr-282095

ABSTRACT

<p><b>OBJECTIVE</b>To investigate therapeutics for and the pathological basis of combined radiation and burn injuries.</p><p><b>METHODS</b>Combined radiation and burn injuries on mice and rats were inflicted by gamma ray irradiation from a (60)Co source and thermal radiation from a 5 kW bromotungsten lamp.</p><p><b>RESULTS</b>The dysfunction of myocardium played an important role in the development of early stage shock. Transfusion of irradiated (in vitro, 20 Gy) or stored (4 degrees C, 7 days) blood after irradiation was done to promote the success of allo-transplantation of bone marrow. Decrease of IL-4 mRNA expression was the molecular basis of depression of intestinal mucosa immune and intervention of IL-4 showed an antagonistic effect on enterogenic infection. A new lipid component extracted from burn eschar was documented for the first time and its toxic effects were elucidated. The survival rate of alloskin grafts after removal of burn eschar from the recipient animals was obviously increased in combined injury due to reduction of immune rejection activity by the radiation effect. In contrast, in animal models with simple burn, the alloskin grafts were all rejected within ten days after the procedure. A successful therapeutic result (survival rate: 92% for 30 days and 67% for 100 days) was obtained by comprehensive management of treated animals, while the untreated control animals all died within 3 - 7 days after injury.</p><p><b>CONCLUSION</b>The pathogenesis of injury caused by simultaneous radiation and burn is extremely complicated and the treatment is very difficult. A comprehensive management program consisting of several therapeutic measures aimed at key links of the pathogenesis may achieve significantly improved results.</p>


Subject(s)
Animals , Mice , Rats , Burns , Pathology , Therapeutics , Calcium , Metabolism , Heart , Hematopoiesis , Radiation Injuries , Pathology , Therapeutics , Rats, Wistar
18.
Article in Chinese | WPRIM | ID: wpr-340942

ABSTRACT

Autologous multipotent stem cells are most relevant cells for regenerative medicine and show prosperous future in the treatment of human diseases. Previous reports have indicated that multipotent stem cells (MSCs) can be obtained from bone marrow and adipose tissues. In this study, we proved that dermis may be another source of these cells. MSCs were isolated from the dermis of newborn rats one day old by adhesion competition and successive culture. These cells conserved the ability to differentiate to osteoblasts, chondrocytes and adipocytes by induction media containing dexamethasone. After long term of more than 6 months, till 25th generation, the cells still maintained the characteristics of stem cells: high activity of self-renewal and multipotency. Mixed collagen matrix from dermis could promote the growth of dermis-derived multipotent stem cells and collagen sponge stent could promote their three-dimensional growth in vitro.


Subject(s)
Animals , Rats , Biomechanical Phenomena , Cell Culture Techniques , Methods , Cells, Cultured , Collagen , Pharmacology , Dermis , Cell Biology , Multipotent Stem Cells , Cell Biology , Physiology , Rats, Wistar
19.
Article in Chinese | WPRIM | ID: wpr-410740

ABSTRACT

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.

20.
Article in Chinese | WPRIM | ID: wpr-410739

ABSTRACT

Objective To study the effects of irradiation and W11-a12,a kind of repair-promoting drug,on anion-selective channel in membranes of mouse peritoneal macrophage. Methods The activity of anion-selective channel was recorded from cell-attached patches with patch clamp techniques. Results The effects of irradiation on anion-selective channel in membranes of peritoneal macrophage included:①decreasing the mean number of activated channels by the presence of zymosan; ②prolonging the mean time from stimulus to the opening of channels; ③depressing the opening of channels by decreasing open-state probability,shortening open-time and prolonging close-time. The effects of irradiation could partly be depressed by W11-a12. Conclusion Irradiation will depress the anion-selective channel of peritoneal macrophage, which may be an important way to depress the function of macrophage.

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