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Objective:To investigate the effect of transforming growth factor (TGF- β) signal in muscle fiber itself during inflammation/immunity response on intramuscular inflammation. Methods:Sixteen wild C57BL/6 mice (wild group) and sixteen mice with skeletal muscle-specific deficiency of T βRⅡ (knock-out group) between 4-8 weeks of age were selected for this study. Acute muscle injury in mice was induced by injection of myotoxin cardiotoxin (CTX) into gastrocnemius. The differences in intramuscular inflammation were compared between the wild and knock-out groups on 0, 4, 7 and 10 d after CTX injection by observing exudation of mononuclear phagocytes, macrophages, M1 type macrophages, CD4 +T cells and helpers T cells (Th1, 2&17). Two newborn C57BL/6 wild mice and 2 SM TGF- βr2-/- knock-out mice were selected to culture primary myoblasts in vitro which were divided into 2 groups: an interferon group subjected to interferon simulation and a control group subjected to addition of an equal amount of solvent. The differences in expression of IL-6, IL-10, MCP-1, MIP-1α, H-2K b, H2-Ea, Toll-like receptor (TLR)3 and TLR7 were compared between the interferon and control groups, as well as between the wild and knock-out groups. Results:On 4&7 d after CTX injection, the ratios of mononuclear/macrophage (75.73%±3.62%, 45.27%± 2.32%), macrophages (38.67%±2.76%, 24.87%±2.19%), M1 macrophages (43.21%±0.11%, 30.43%±2.19%), CD4 +T cells (20.13%±1.62%, 5.67%±0.32%) in the muscle tissue from the knock-out mice were significantly higher than those from the wild mice (58.52%±2.43%, 29.21%±2.45%; 20.63%±2.32%, 16.23%±1.25%; 24.98%±0.35%, 14.23%±1.69%; 10.70%±0.43%, 2.50%±0.45%), with a majority of Th1&Th17 ( P<0.05). In vitro results showed that the levels of IL-6, MCP-1, MIP-1α, H-2K b, H2-Ea and TLR3 were significantly upregulated in the interferon group compared with the control group and that such upregulation in the nock-out mice was more significant than in the wild mice ( P<0.05). Conclusions:Endogenous TGF- β signal activation plays a role in the functional recovery after muscle trauma, because it is involved in the regulation of immune behavior of muscle fibers, thus affecting intramuscular inflammation and muscle regeneration.
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Objective To measure 5Fu concentration after intraoperative regional arterial perfusion chemotherapy in gastric antrum cancer patients.Method In this study,32 gastric antrum cancer patients intraoperatively received through right gastroepiploic artery intraarterial perfusion of 5-Fu (1000 mg/m2),and drug conentrations were measured in portal venous blood,peripheral venous blood,peritoneal fluid and in removed cancer tissues 2,5,10,20,30,60 minutes after infusion started by high performance liquid chromatography method.Result 5-Fu in portal vein blood and peritoneal fluid reached peak concentration after 2 minutes at (48.8 ±6.8) μg1/ml and (75.3 ±30.7) μg/ml,respectively.The drug concentration in portal venous blood was (19.2 ± 2.0) μg/ml at 30 minutes and that in peritoneal fluid was (17.3 ±7.4) μg/ml at 60 minutes,both were higher than that of the effective experimental tumor suppressing concentration (15 μg/ml).Peripheral venous blood concentration reached peak concentration of (5.4 ± 2.0) μg/ml at 5 minute.The drug concentration in removed cancer tissues was (80.5 ± 20.1) μg,/ml.Conclusions Regional intraarterial perfusion chemotherapeutics in gastric antrum cancer patients during operation immediately makes the concentration of chemotherapy rise in the portal vein blood and peritoneal fluid and in cancer tissues to a much higher level than that in peripheral blood.This helps kill cancer cells that fell out during the surgery.
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Objective To study the transfection effects of nuclear factor-KappaB(NF-κB)decoy oligodeoxynucleotides(ODN) to Kupffer cells (KCs) mediated by lipofectamine,and investigate it's suppression effects on KCs activation. Methods Twenty-four Wistar rats were divided into three groups (n=8).(1)Control group,in which the normal KCs were isolated.(2)LPS group,in which 1 ms/L LPs was added to the culture system.(3)NF-κB decoy ODN group,in which KCs were transduced with NF-κB decoy ODN (4μg×105KCs)prior to LPS stimulation.The transfection efficiency Was assayed,and the phagocytosis function,NF-κB(P65) translocation,CD40 mRNA expression of KCs were also detected respectively. Results Kupffer cells were obviously activated after LPS stimulation.the phagocytosis function was reinforced.the activity of NF-κB transloeated from cytoplasm into nucleus was obviosly increaced.The co-stimulatory molecules expression(CD40 mRNA)significantly increased compared with control group(t=4.01,P<0.01).NF-κB decoy oligodeoxynucleotides can efficiently transfected into KCs mediated by lipofectamine,which can obviously suppress KCs activation,and downregulate the expression of downstream gene(compared with LPS group,t=4.89,P<0.01). Condusion NF-κB decoy ODN can efficiently transfect into KCs and inhibit it's activation.
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BACKGROUND: Heme oxygenase-1 (HO-1) plays an important role on preventing tissues and organs from oxidant stress injury, which remains currently one of the most active areas of investigation. OBJECTIVE: To study HO-1 over-expression on the survival and fiver function of rats after reduced-size liver transplantation by constructing recombinant adenovims Ad5-HO-1.DESIGN, TIME AND SETTING: Randomized controlled animal study was performed in the Institute of Pediatrics, Children Hospital Affiliated to Chongqing Medical University from September 2003 to March 2005. MATERIALS: Seventy-four SD rats were used to establish in situ reduced-sized liver transplantation models. METHODS: Recombinant adenoviral vector encoding Ha-1 gene (AdS-HO-1) was generatred by molecular biology method, wluch was administered to donors via voln at 48 hours before transplantation. All rats were randomly divided into control group(n=12),saline group(n=12), cobalt protoporphyrin (CoPP)group(n=13),Ad5-HO-1 group(n=13),Ad5-green fluorescent protein(GFP)group(n=12),and Ad5-HO-1+zinc protoporphyrin (ZnPP)group (n=12).Livers of donor wefe harvested and stored for 24 hours at 4℃ in HTK solution. Before it is implantedinto recipient. MA IN OUTCOME MEASURES: Survival rate and liver function; portal vein blood flow monitored 2 hours after transplantation by Color Doppler How Imaging; pathological changes of transplanted liver tissue observed by HE staining;HO-1 activity, tumor necrosis factor-alpha(TNF-α),Bcl-2 and Bax expression detected by immunohistoehemical staining; changes of HO-1,TNF-α,bcl-2 and bax mRNA detected by molecular viewpoint. RESULTS: Survival rate in the Ad5-HO-1 group was significantly higher than that in the saline group at al,7,and 21 days after liver transplantation(P＜0.05).Glutamic pyruvic transaminase decreased in the Ad5-HO-1 group as compared to that in the saline,Ad5-GFP,and Ad5-HO-1+ZnPP groups(P＜0.05);portal venous blood flow significantly increased 2 hours after transplantation(P＜0.05);HO-1 activity also significantly increased(P＜0.05).RT-PCR and immunohistochemical staining showed that HO-1 and bcl-2 expressions increased(P＜0.05),but bax and TNF-αexpressions decreased(P＜0.05).CONCLUSION:Ad5-HO-1 significantly induces high expression of HO-1 increases portal venous blood flow within 2 hours alter liver transplantation, and promotes liver functional recovery so as to prolong survival time of rats after liver transplantion.
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Objective To clarify great benefit to prevention of wound infection as well as reduction of economic expenditures by a new method of connecting latex gloves and cigarettes in peritoneal drainage. Methods We randomly selected 121 cases of patients with peritoneal pyogenic infection, divided them into the latex gloves group (64 eases)and the common dressing group (57 cases). The incision infection rate and eco-nomic cost in two groups were comparately studied. Results The latex gloves group's incision infection rate was 10.94% (7 / 64), the common dressing was 31.58% (18/57). Each patient of the latex gloves group spent less cost about 15.70 yuan,changed dressing 5.3 times less than the common dressing group. Conclusions Connecting latex gloves and cigarettes in peritoneal drainage is a new method of reducing economic expandi-tures and the chances of incision infection. Clinical application of this method should be fully prospected.
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Objective:To establish the acute rejection model of orthotopic liver transplantation in rat(ROLT) and observe the basic pathophysiologic changes of acute rejection.Methods:The rats were randomly divided into three groups:control group,isotransplantation group(LEW-LEW),and allotransplantation group(LEW-BN).Recipients were sacrificed on 3,5,7,and 10 days postoperatively and liver tissues and blood samples were collected.Recipient survival rate,histopathological and ultrastructural characteristics were observed.ALT,TBIL,and Alb were measured with automatic biochemical analyser.IL-2 content in serum was assayed by ELISA.Results:There was no rejection in isotransplantation from Lewis to Lewis rat resulted and 14-day survival rate reached 100%.The IL-2 concentration in serum was at normal level.On the contrary,all recipients in allotransplantation group from LEW to BN rat died among 14 days postoperatively,and hepatic histological examination showed typical acute rejection on 7 days.Liver function was severely impaired,which was indicated by significant increase of ALT and TBIL levels and apparent decrease of Alb level.The IL-2 concentration in serum was continuously increased and reached its peak value on 7 days postoperatively.Conclusion:An acute rejection experimental model of liver transplantation in rat could be stably established using Lewis rat as donor and BN rat as recipient.