【Abstract】Objective To observe the synthesis of TLR2 protein and its mRNA expression in Kupffer cells (KCs) and sinusoidal endothelial cells(SECs).Methods Thirty-two BALB/c mice divided into two groups (operation group and false operation group) were used to prepare the model of partial hepatic ischemia/reperfusion (I/R) injury. After injury KCs and SECs were isolated with twosteps situ perfusion technique. And these cells were dyed by rat anti-mouse TLR2 IgG and anti-rat IgG2b labeled with flurescein isothiocyanate (FITC). The sysnthesis of TLR2 protein were determined by flow cytometric (FCM) analysis and real time reverse transcription polymerase chain reaction (Real-Time RT-PCR) analysis for gene expression.Results As for KCs: TLR2 expression was significant higher in operation group, compared with false operation group 〔protein expression: （9.19±1.07）% vs （1.52±0.21）%, P＜0.01; gene expression: 0.54±0.77 vs 2.62±2.19, P＜0.05〕. But there were no significant differences with expression in SECs. Conclusion Synthesis of TLR2 protein and its gene expression increased in KCs in the mouse partial hepatic ischemia-reperfusion injury.
The canine saphaneous skin flap was used as a model in this experiment. The cutaneous autograft would give long-term survival, whereas the allograft without pretreatment would only survive 10. 2±1.9 days from its transplantation. If the pretreatment consisted of the use of immunosuppressive agent as PHA or infusion of dexamesone, the survival days of the allografts could be prolonged to 15.1±2.5 and 13.7±2.8, respectively(Plt;0.01). The histological examination gave the evidence that drug perfusion delayed the rejection.
Objective To study the effect of Kupffer cell on the liver ischemia/reperfusion injury.Methods The literature in recent years on the liver ischemia/reperfusin injury were reviewed.Results The activated kupffer cell can generate and release a variety of soluble toxic mediators, affect the liver microcirculation directly or indirectly. Conclusion Kupffer cell have important effect on liver ischemia/reperfusion injury.
To investigate the protective effect of propofol on ischemia/reperfusion induced spinal cord injury in rabbits and its influence on excitatory amino acid (EAA). Methods Sixty New Zealand white rabbits weighing 2.0-2.5 kg, half males and half females, were selected. The infrarenal circumaortic clamping model was used. And 6 mL/kg different fluids were continuously infused through a catheter into the aorta distal to the clamping site at a speed of 12 mL/(kg?h) during the 30 minutes ischemia period. According to the different infusing l iquids, the rabbits were randomized into 6 groups(n=10 per group): group A, normal sal ine; group B, 10% intral ipid; group C, propofol 30 mg/kg; group D, propofol 40 mg/kg; group E, propofol 50 mg/kg; group F, propofol 60 mg/kg. At 0, 6, 24, and 48 hours after reperfusion, neurologic outcomes were scored on a Tarlov scale system. At 48 hours after reperfusion, the number of normal neurons in the anterior spinal cord was counted, and concentration of EAA in the lumbar spinal cord was measured by high performance l iquid chromatography. Results The neuroethological score was better in groups C, D, E and F than that of groups A and B (P lt; 0.05), the score of group E was the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). The number of normal neurons in the anterior spinal cord of groups C, D, E and F was greater than that of groups A and B (P lt; 0.05), and group E was greater than groups C, D and F (P lt; 0.05). The concentration of EAA in groups A, B, C, D, E and F was greater than that of normal tissue, the group E was the lowest (P lt; 0.05), the groups A and B were the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). Concentrations of glutamate and aspartic acid were negatively correlated to normal neuron numbers in the anterior spinal cord and neuroethological scores 48 hours after reperfusion, and the corresponding correlation coefficient was — 0.613, — 0.536, — 0.874 and — 0.813, respectively (P lt; 0.01). Conclusion Propofol can significantly inhibit the accumulation of EAA in spinal cord and provide a protective effect against the ischemia/reperfusion injury induced spinal cord in rabbits.
Objective To investigate the effects on myocardial perfusion of simultaneous antegrade/retrograde cardiopiegia （SARC） through a single coronary artery and coronary sinus （CS）. Methods SARC was conducted in isolated pig hearts through CS in conjunction with the left anterior descending （LAD）, the left circumflex （LCX）, or the right coronary artery （RCA） respectively. After injecting magnetic resonance （MR） contrast agent （gadolinium diethyienetriamine pentaacetic acid, Gd-DTPA） into arterial or venous route, the distribution of Gd-DTPA with magnetic resonance imaging（MRI） was monitored and the effluent from the venting coronary arteries to assess the efficacy of SARC for myocardial perfusion was measured. Results Injection of Gd-DTPA into a perfusing artery during SARC resulted in increased signal intensity not only in the territory of the perfusing artery but also in the areas normally served by the other two venting coronary arteries （including the right ventrieuiar free wall）. With Gd DTPA given into the CS during SARC, the myocardium in the territories of the two venting coronary arteries was lightened. Signal intensity of the myocardium in the perfusing artery territory and right ventricuiar free wall remained unchanged. Moreover, a significant amount of effluent was collected from the venting coronary arteries during SARC： the LAD 10.5-17.7 ml/min; LCX 9.7-15.2 ml/min, and RCA 4.7-7.8 ml/min. Conclusion SARC through a single coronary artery and CS can provide homogeneous perfusion to the entire heart and is sufficient to prevent ischemic injury in the myocardium normally supported by the venting coronary arteries.
ObjectiveTo evaluate the protective effect of estrogen on survival of retinal ganglion cells (RGCs) after transient retinal ischemia-reperfusion (RIR) in rats.MethodsRIR was induced in 60 ovariectomized adult rats (OVX) by increasing intraocular pressure via an intracameral catheter. All of the rats were divided into two groups randomly: in experimental group, the rats underwent a subcutaneous injection with 17β-estrodiol（100 μg/kg） 2 hours before retinal ischemia; and in the control group, saline water was injected correspondingly. The number of RGCs and the thickness of the inner retinal layers were mesured by HE staining method before and 12, 24, 48, and 72 hours after reperfusion. TdT-mediated biotin-dUTP nick end labelling (TUNEL) staining technique was used to examine the apoptosis of RGCs.ResultsTwenty-four and 48 hours after reperfusion, the number of apoptotic cells in experimental group was obvious lower than that in the control group(Plt;0.05), and the number of RGCs in experimental group was higher than that in the control group(Plt;0.05).ConclusionEstrogen can protect retinal neurons from transient RIR in ovariectomized rats.(Chin J Ocul Fundus Dis, 2005,21:177-179)
OBJECTIVE: To investigate the injury on isolated testes induced by ischemia/reperfusion(I/R), and the protective effect of Yisheng injection on the injury. METHODS: Twenty-six isolated cadaver testes contributed by 13 persons were preserved with 4 degrees C 250 ml hypertonic citrate alloxuric (HCA) solution and then reperfused with 37 degrees C 500 ml HCA. Solution of experimental group contained 500 micrograms/ml Yisheng injection. In simple cold preservation test, involving in 8 experimental and 8 control testes, a series of time points (6, 12, 18, 24, 36, 48, 60, 72 hours) were set to harvest. 10 testes (1 testis respectively on 6, 12, 18, 24 and 36 hours in experimental and control groups) were reperfused with 37 degrees C HCA for 6 and 12 hours. Histological and histochemical changes were observed. RESULTS: In the experimental testes, 4 degrees C cold preservation in 24 hours could not induce obvious pathologic changes. After 24 hours, changes such as swelling, vacuolar degeneration or detachment of endothelial cells (ECs), separation between basement membrane and seminiferous epithelium, mal-alignment of spermatogenous cell and edema of mesenchyme could be observed. In the testes preserved for 12 hours, the activity of lactic dehydrogenase(LDH) and succinic dehydrogenase (SDH) increased, then fallen after 24 hours. The activity of Nitric oxide synthetase(NOS) decreased after 18 hours. All changes were more obvious after following 37 degrees C reperfusion. In the control testes, swelling and vacuolar degeneration of ECs occurred on 12 hours cold preservation, and injury was worse along with the prolongation of cold preservation time. Pathologic changes of ECs, seminiferous epithelium and mesenchyme were serious after 37 degrees C reperfusion. CONCLUSION: 4 degrees C cold preservation in 24 hours can only cause mild ECs’ injury, and obvious abnormal testes’ histological profile can be observed beyond 24 hours. 37 degrees C reperfusion will make injury worse. Yisheng injection can keep isolated testes histologic structure well in 24 hours cold preservation, and it has protective effect on I/R injury.
Objective To investigate the extent of hepatic ischemia reperfusion (HIR) injury in rat cirrhotic liver under different ischemic time，and find the time limit under which the rat with cirrhotic liver could tolerate. Methods At first，the cirrhosis of the rat were induced by carbon tetrachloride（CCl4）injected subcutaneously. Then these rats were randomly divided into four groups. Group A(n=6) was made by sham operation, group B, C, D(n=16) were respectively given 20, 30, 40min hepatic warm ischemia. The 7day survival rate, AST, ALT, TNF and liver, pulmonary pathology were observed. Results The 7-day survival rate was decreased with the increase of hepatic ischemic time. The survival rate of group B, C, D were respectively 100%, 60%, 40%. Between group C, D and group B there were significant differences（P＜0.05）. The level of AST and ALT in group D were （2 448.4±942.3）u/L and （1 189.0±403.4）u/L respectively, and those in group C were （2 185.1±1 732.9）u/L and （1 183.5±707.2）u/L respectively, which were higher than those in group B and A significantly（P＜0.01）. The level of TNF was increased significantly 4hr after reperfusion, as compared with that before operation 〔(0.177±0.139）u/ml〕, P＜0.01. TNF of group B, C, D were （0.399±0.216）u/ml, （0.671±0.351）u/ml and （0.789±0.371）u/ml respectively. At the same time the level of TNF in group C, D was higher than that in group B, A significantly（P＜0.01）. Liver and lung pathology showed increased damage with increasing ischemia. Conclusion Hepatic injury is induced by HIR in rats with cirrhotic liver, and its severity increases with the increase of ischemic time. There is a certain hepatic ischemic time between 20min and 30min, which can be tolerated by the rats with cirrhotic liver. TNF may be used as an indicator，showing the degree of HIR injury and foreseeing the result of injury.
摘要:目的:探討晚期食管癌切除、縱隔淋巴結清掃及術中縱隔熱灌注化療對殘留于氣管、支氣管、胸主動脈、奇靜脈等器官的癌性肉眼微小病灶治療效果。方法:選擇食管癌病變浸潤超過外膜層外侵至氣管、支氣管、胸主動脈、奇靜脈等器官患者112例，隨機分為兩組:治療組56例,術中42～43℃無菌蒸餾水2000～2500 mL加入順鉑(DDP)150 mg及氟尿嘧啶(5FU)1200 mg在體外循環下行縱隔熱灌注化療40 min;對照組56例,術中未進行縱隔熱灌注化療。結果:治療組術后第一年有6例出現縱隔區域腫瘤復發及淋巴結轉移,術后第二年有11例縱隔區域腫瘤復發及淋巴結轉移;對照組術后第一年有14例出現縱隔區域腫瘤復發及淋巴結轉移,術后第二年23例出現縱隔區域腫瘤復發及淋巴結轉移。結論:晚期食管癌術中縱隔熱灌注化療可明顯減少或延遲縱隔區域腫瘤復發及淋巴轉移,提高術后第一至第二年生存率。Abstract: Objective: To explore the advanced esophageal cancer resection, mediastinum, lymph node dissection, mediastinum, hot infusion chemoembolization and clinical observation of residual heat infusion chemoembolization and trachea, or the thoracic aorta, bronchus, eye cancer organs such as intravenous of tiny lesions therapeutic effect. Methods: Select esophageal lesions than the outer membrane layer of infiltrating the trachea and bronchus to the thoracic aorta, and 112 cases of patients with venous organs such as random points to two groups: treatment group treated with perfusion of 56 cases at 4243 degrees Celsius sterile 2000 mL distilled water 2500 mL ~ (DDP) joined cisplatin 150 mg, 5fluorouracil (5FU 1200 mg) in extracorporeal circulation downlink mediastinal hot perfusion 40 minutes, control group treated with perfusion of 56 cases without mediastinal hot perfusion chemotherapy. Results: Treatment group in 6 cases occured after first mediastinal tumor recurrence and regional lymph node metastases after 11 cases, the regional recurrence and lymphatic metastasis mediastinal, control group first fill after 14 cases mediastinal tumor recurrence and bureau of regional lymph node metastasis appeared in 23 cases, surgery between regional tumor locally recurrent lymph node metastases. Conclusion: Advanced esophageal intraoperative mediastinal hot perfusion chemotherapy can obviously reduce or delay mediastinal tumor recurrence and regional lymph node metastases, raise the firstsurial.
ObjectiveTo observe the effect of Crocin on structure and the expression of tumor necrosis factor-alpha; (TNF-alpha;) and interleukin-1beta; (IL-1beta;) in rat retina after injury by ischemia-reperfusion. Methods A total of 80 Sprague-Dawley male rats at the age of 8 -10 weeks were divided into control group, model group, low-dose Crocin group and high-dose Crocin group, with 20 rats in each group. The rats of control group were not treated. The rats in model, low-dose Crocin and high-dose Crocin group were induced with normal saline by anterior chamber perfusion creating a retinal ischemia-reperfusion (RIR) model. The rats of the low-dose Crocin and highdose Crocin group received intraperitoneal injection with different doses of Crocin solution (5 mg/kg, or 50 mg/kg) 30 minutes prior to ischemic injury and one time per day after successful RIR. Optical microscopy was used to observe the retinal structure. Enzymelinked immunosorbent assay (ELISA) was used to measure the expression of TNF-alpha; and IL-1beta; 6, 12, 24 and 48 hours after RIR. ResultsThe retinal structure of control group was normal. Pathological changes were found in the RIR model and low-dose Crocin group, such as retinal edema, disorganized structure and loosely packed cells. The degree of pathological changes in lowdose Crocin group was less than the RIR model group. The retinal structure of high-dose Crocin group was similar to the control group. The expression of TNF-alpha; was the highest at 24 hours after modeling, while the expression of IL-1beta; was the highest at 12 and 48 hours after RIR modeling. Six, 12, 24 and 48 hours after RIR modeling, compared with the control group, the TNF-alpha; expression of model (t=5.42, 7.94, 9.32, 9.18;P＜0.05 ), low-dose Crocin (t=3.94, 4.12, 4.98, 3.84;P＜0.05) and high-dose Crocin group (t=2.13, 2.34, 2.96, 2.78;P＞0.05) were increased. Compared with the RIR model group, the TNF-alpha; expression of low-dose Crocin (t=3.95, 4.56, 4.01, 5.12) and high-dose Crocin group (t=5.23, 7.65, 7.74, 7.63) was decreased. Compared with the control group, the IL-1beta; expression of model (t=7.23, 7.87, 7.15, 15.60), low-dose Crocin (t=5.65, 5.10, 5.54, 6.87;P＜0.05) and high-dose Crocin group (t=4.38, 5.21, 4.56, 4.75) was increased (P＜0.05). Compared with the model group, the IL-1beta; expression of low.dose Crocin group was decreased significantly 48 hours after RIR modeling (t=7.56,P＜0.05); but it decreased significantly at each time point in high-dose Crocin group (t=6.94, 5.36, 6.05, 10.50;P＜0.05). Conclusion Crocin can improve the retinal pathologic changes, while down-regulating TNF-alpha; and IL-1beta; expression in RIR rats.