ABSTRACT
Objective To investigate the effects of G-CSF-mobilized autologous stem cells in the prevention of radiation pulmonary injury.Methods Mice were divided into control group,irradiation group and treatment group.Mouse model of pulmonary fibrosis was established by exposing chest to a single dose of 14 Gy.Animals in the treatment group received recombinant human G-CSF (250 μg/kg daily for 5 d) before the irradiation in order to mobilize autologous stem cells in vivo.The general condition and mortality were documented after radiation injury.The pathological study with histological scoring,Masson staining and Sirius red staining with polarized light analysis were used to identify lung injury and the potential benefit of stem cell mobilization.Results Local chest irradiation of a single dose of 14 Gy was a suitable dose to create radiation-induced pulmonary fibrosis in mice.The death rate was 37.5％,which mainly happened around 11 weeks after injury.In contrast,all of the animals in G-CSF treated group survived.The ratio of lung to body mass was significantly increased in both irradiation group and treatment group (F =23.20,P＜0.05) around 3 months after the injury,with a higher ratio in irradiation group than that in treatment group (P＜0.05).Histological scoring for alveolar inflammation at 3 months after injury revealed statistically significant difference in irradiation group and treatment group compared with control group (F=11.93,P＜ 0.05).At this time point,the pathological observation showed lung tissue degeneration and necrosis with alveolitis and interstitial inflammation,as well as fibroblasts proliferation and focal collagen deposition in alveolar septa.At 4 month after the injury,the inflammation ininterstitial tissue was receded,but fibrosis and collagen deposition were significantly increased.In addition,at 3 and 4 months afterinjury,the pulmonary fibrosis was aggravated in irradiation group (F=28.73,16.85,P＜0.05),and significantly alleviated in the treatment group (P＜0.05).The similar results were confirmed in collagen content analysis (IOD) by Sirius red staining and image analysis (F =17.70,17.79,P＜ 0.05).Conclusions Autologous mobilization of stem cells could prevent the death of radiation-injured animals possibly by alleviating early lung injury and interstitial inflammation as well as the late pulmonary fibrosis,suggesting a therapeutic potential of autologous stem cell mobilization in radiation pulmonary fibrosis.
ABSTRACT
Combined radiation-trauma injury is mainly observed in radiation treatment of cancer and radiation injury with traumatic patients. The prominent problem of combined radiation-trauma injury is delayed or prolonged wound healing. The mechanism of the impaired wound healing is complicated, and the current effective treatment method are limited. This paper reviews the mechanism and treatment of this impaired wound healing, including the cellular depletion, stromal cell dysfunction, aberrant collagen deposition, microvascular damage, as well as the targeted therapies for the impaired wound healing such as stem cell repletion, antioxidant therapy, transforming growth factor beta-1 ( TGF-β1 ) modulation, and implantable biomaterials. This paper is designed to provide a reference for further deep research on the mechanism and treatment of radiation-trauma injury.
ABSTRACT
Objective To explore characteristics and significance of interferon-induced protein with tetratricopetide repeats 1 expressed in radiation injury and infection stress.Methods RNA was extracted from Raw264.7 cell,3T3 cell and 10T1 /2 cell after 5 hours stimulated with 5 μg/mL LPS.At the same time,to set up normal control group (untreated by LPS),and RNA of IFIT1 was detected by RT-PCR.Total-ly 20 C57 /BL6 mice were randomly divided into 4 groups,namely 0 h group,1 h group,4 h group and 12 h group.The mice were given 12 Gray60Co full-body exposure once,then liver IFIT1 was detected by western blot.Results Stimulated with LPS for 5 hours,IFIT1 was in-duced expression in Raw264.7 cell,3T3 cell and 10T1 /2 cell.The expression of normal control group was negative.The level of IFIT1 /Actin increased significantly 1 hour after radiation injury,and it reached the peak 12 hours after radiation injury (P <0.01).Conclusion LPS can stimulated a variety of cell lines expressed IFIT1,prompting that IFIT1 may participate in the occurrence and development of post-traumatic toxemia.IFIT1 of liver tissue increased significantly during the early stage in radiation mice.
ABSTRACT
Objective To explore the changes of gut microbiota in response to abdominal and pelvic radiotherapy and its potential relationship with intestinal infection.Methods Irradiation was delivered to the abdominal region of BALB/c mice,following the regular human pelvic-radiotherapy protocol,2.0 Gy/d,continuous 5 d/week.Samples of ileum tissue and the intestinal content were collected at different time points of irradiation procedure,including after 3 and 5 weeks,and at 1 week after 6 weeks of irradiation.Quantitative RT-PCR was used to measure the mRNA level of antimicrobial peptides and pro-inflammtory factors.Bacterial translocation was determined by PCR.The gut microbiota was characterized by the denaturing gradient electrophoresis assay.Results The expressions of cryptdin-1 and cryptdin-4 were decreased after 3 weeks of irradiation and at 1 week after 6 weeks of irradiation(t =-7.43,-3.54,-4.72,-4.27,P ＜ 0.05),while they were significantly increased at the 5 weeks of radiation (t =6.15,5.75,P ＜ 0.05).The diversity index and richness of gut microbiota after 3 or 5 weeks irradiation were significantly decreased (t =-3.49,-4.19,-3.44,-4.97,P ＜ 0.05).The gut microbiota dysbiosis of the irradiated mice was characterized with the decrease of probiotics of Lactobacillus and the increasing of opportunistic pathogen of Escherichia coli,Shigella flexneri,et al.Bacterial translocation episodes were more frequently in the irradiated mice than that of control animal.The mRNA levels of IL-1β、IL-6 and TNF-α were significantly increased after 3 or 5 weeks of irradiation (t =4.85,6.16,7.71,4.60,4.86,5.97,P ＜ 0.05).Compared with the control,the expression levels of IL-1β and TNF-α at the 1 week after 6 weeks of irradiation ending was also obviously enhanced (t =3.67,5.88,P ＜0.05).Conclusions Pelvic radiotherapy can induce abnormality of enteric antimicrobial peptides and may result in gut microbiota dysbiosis.The disturbed gut microbial flora may further trigger an incurrence of bacterial translocation and enteritis.Therefore,the gut microbiota may be a potential interfering target to alleviate radiotherapy adverse effect.
ABSTRACT
Objective To construct the eukaryotic expression vector for mouse microRNA miR-21 and identification its expression activ-ity in 293 cells. Methods The genomic sequence containing pre-miR-21 was amplified from mouse genomic DNA by PCR and cloned into the pRC/CMV plasmid. The constructed recombinant plasmid pRC/CMV-mmu-miR-21 was transfected to 293 cells by lipofectamine 2000, and the stably transfected cells were screened with G418,from which total RNA was extracted for detecting the expression of mature miR-21 by northern blot. In the meantime,a luciferase report plasmid examing the activity of miR-21 named pmiR-21-Luc reporter was also construc-ted,and luciferase activity analysis indicated the product of pRC/CMV-mmu-miR-21 indeed had biological activity. Results Both restriction enzyme digestion analysis and sequencing proved the recombinant plasmids were constructed correctly. The miR-21 was highly expressed in the screened clones of 293 cells and it had good biological activity. Conclusion The eukaryotic expression plasmid of mouse miR-21 was successfully constructed,which laid the foundation of further investigation of the role of miR-21 during skin wound healing.
ABSTRACT
Objective To assess the distribution of recombinant fusion protein dTMP-GH in mice and to determine whether it is of tar-geted distribution characteristics. Methods A laboratory scale preparation of dTMP-GH recombinant fusion protein was obtained. Protein dT-MP-GH was labeled with radioactive 125 I,then mice were sacrificed at 5 min,15 min,30 min,1 h,2 h,4 h,8 h,12 h,24 h after tail vein injec-tion of 125 I-dTMP-GH at a dose of 100 μg/kg,and the organs and tissues ( heart,liver,spleen,kidney,bone and thyroid) were collected for radioactive counting. Results Preparation of purified ( >98%) dTMP-G was obtained. 125 I labeling rate was 71. 53%,radiochemical purity was 96. 53%,and specific activity was 0. 22 MBq/μl. 30 min after tail vein injection of 125 I labeled dTMP-GH,radioactivity accounted for 10% of the total injected in femoral,and metabolism was carried via liver and kidney over time. Conclusion Fusion protein mainly distribu-ted in bone marrow via tail vein injection in mice,which expressed that dTMP-GH has the characteristics of selective distribution in bone mar-row tissue.
ABSTRACT
Upper urinary tract damage secondary to voiding dysfunction is an important reason for the end stage of renal diseases. We evaluated clinical manifestations and outcomes of various treatments, and analyzed underlying mechanism in order to improve guidance for their therapy. Two hundred seventy one patients suffering from upper urinary tract damage caused by voiding dysfunction diseases from Jan. 1999 to Oct. 2009 were enrolled and data of urodynamics, urinary imaging and renal functions before and after treatments were retrospectively analyzed. Bladder pressure was over 40 cm H[2]O in 78.2% of the patients, and among them, the average pressures for the first voiding desire was 42.3 +/- 6.0 cm H[2]O with the maximum bladder pressure of 67.3 +/- 5.8cmH[2]O in 271 patients. In 157 patients the residual urine bladder volume was 100ml, but smaller and moderate residual urine volume was occurring in 37.9% of the upper urinary tract damage patients, which implied that residual urine is not an appropriate indicator. 89.3% of the patients exhibited bladder outlet obstruction and 95.2% of those suffered moderate to severe LUTS. After the relief of obstruction treatment and resulting intravesical pressure reduction, 264 patients showed a good recovery and in 202 of them hydronephrosis have disappeared and their renal function returned to normal, due to reduced bladder pressure. We found that high bladder pressure is the main reason of upper urinary tract damage caused by voiding dysfunction. Thus, attention should be paid in order to achieve and maintain reduced bladder pressure below the safety line
ABSTRACT
Objective To study the relationship of expression of central cortex glucocorticoid receptor (GR) at protein level with GR expression in the liver at protein level and with changes of serum cortisol and adrenocorticotropic hormone (ACTH) following severe closed traumatic brain injury (TBI) in mice. Methods Severe TBI was established in awake mice by using a BIM-Ⅲ biomechanical machine. At 0.5, 2, 8, 24, 48 and 72 hours after TBI, the total cytosolic GR in the cortex and liver were detected with Western blotting. Levels of serum ACTH and cortisol were measured by ELISA technique and radio-immunological assay (RIA) respectively. Results The expression of GR both in the cortex and liver were obviously down-regulated at protein levels at 2-72 hours after TBI and increased slowly eight hours after injury. The GR in the liver showed no recovery at 72 hours after injury and that in the cortex was decreased continually at 24 hours after injury. Serum ACTH and cortisol levels were increased markedly compared with control group, when there were two different peaks in the observation curve.Conclusion There is glucocorticoid resistance both in the central and peripheral tissues after severe closed TBI in the awake mice, which changes in a time-dependent manner.
ABSTRACT
Objective To investigate early changes of angiopoietin-2(Ang-2)in multiple trauma patients and assess its clinical significance.Methods Forty-five multiple trauma patients aged 2060 years admitted to the hospital within one hour after injury were randomly divided into three groups according to injury severity score(ISS).Blood specimens were obtained immediately upon arrival in the emergency department and plasma samples were assayed for comparing changes of Ang-2,TNF-a and IL-6.Meanwhile,plasma level of Ang-2 was measured and analyzed under different oxygenation index,shock index and base deficit.Results Plasma level of Ang-2 was positively correlated with ISS(P ＜ 0.01)and was concordant with the plasma levels of TNF-a and IL-6(P＜0.01).Furthermore,plasma level of Ang-2 was elevated upon increase of shock index or decrease of oxygenation index(P ＜ 0.01).Plasma level of Ang-2 was elevated with the increase of base deficit(P ＜ 0.01).Conclusions High level of Ang-2 is a marker of endothelial activation and dysfunction early after trauma.Ang-2 is related tightly with the injury severity,inflammation factors,systemic oxygenation and tissue hypoperfusion and may have a tight relation with pathophysiological development and clinical outcome after trauma.
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.
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.
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 , GeneticsABSTRACT
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.
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.
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.
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.
ABSTRACT
Objective To investigate the changes of the expression of HSP90? in the liver of severely scalded rats at the early stage of stress Methods The expression levels of HSP90? in the liver of rats at different time points after severe scald were detected by Western blotting and then the expression of HSP90? in the nucleuses was observed by immunohistochemistry Results The expression of HSP90? significantly increased in the liver of rats after severe scald, especially during 12～48 h Abnormal high expression of HSP90? in the nucleuses around was found at 48 h after scald Conclusion The abnormal high expression of HSP90? due to severe scald may be one of the important causes resulting in the disorder of glucocorticoid receptor at the early stage of stress after severe trauma.
ABSTRACT
Objective To review the characteristics of human defensin 5 (HD 5), including molecular structure, antibacterial activity and gene expression, and to show its development prospect as a new drug in the treatment of enterogenic infection. Methods The published papers about HD 5 were reviewed to summarize its research progress. Results Being a 3-5 KDa cationic peptide rich in cysteine and arginine, especially without glycosylated side chain, HD 5 plays its antibacterial role against positive and negative Gram's bacterium, fungi, spirochete, protozoan and enveloped virus with the special active center composed of three disulfide bonds. HD 5 encoding gene alpha defensin 5 (DEFA 5) localizes at the 8th chromosome P21 pter with 449 bp, which includes five pieces of sequence: 5′ untranslated region (1-40),signal peptide (41-97), propiece (98-226), mature peptide (227-322),and Poly A (433-438).Conclusion As a broad spectrum and effective endogenous antimicrobial peptide, HD 5 would be a promising alternative peptide against enterogenic infection if the accessibility to its mass production is settled.
ABSTRACT
Objective To study the role of Cysteine-rich 61 ( Cyr61/CNN1) in repair of combined injury by observing whether exogenous CCN1 promotes the proliferation and migration of radiation-injured L929 cells. Methods A radiation model of L929 cells was induced by ? ray at a dose of 4 Gy. The irradiated L929 cells were cultured in a medium containing 2 ml adenovirus plasmids of CCN1 or RFP at 37 ℃ in an atmosphere containing 5% CO2,which served as a CCN1 group and a RFP group,respectively. Irradiated L929 cells cultured in a blank control medium served as a blank control group. Effects of CCN1 on proliferation and migration of radiation-injured L929 cells were detected by MTT assay,plate colony formation assay,cell cycle analysis,and scratch test of wound healing. Results The proliferation and colony formation rates of L929 cells cultured in a medium containing CCN1 were significantly higher than in RFP and control groups [colony formation rate:( 34. 4 ?3. 6) % vs ( 24. 5 ?2. 9) % and ( 29. 5 ?3. 5) %,P
ABSTRACT
Objective To fulfill high-efficient expression of rTMP-GH,a recombinant fusion protein of thrombopoietin mimetic peptide and human growth hormone,in Pichia pastoris.Methods cDNA fragment coding for rTMP-GH was amplified by PCR,and subsequently cloned into the expression vector pPIC9K.The linearlized plasmid pPIC9K-rTMP-GH was transformed into Pichia pastoris GS115 cells and screened in MD/MM plates under pressure of G418.The recombinant fusion protein rTMP-GH was identified by Western blotting,and its biological activity was analyzed by its ability on colony formation of megakaryoblasts.Results The pPIC9K-rTMP-GH expression plasmid was successfully constructed.GS115 cells with high expression of rTMP-GH was screened out,and the rough purified rTMP-GH showed promotive effect on megakaryoblast colony formation.Conclusion High-efficient expression of rTMP-GH in Pichia pastoris is achieved.