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AIM: To investigate the effects of 0.01% atropine eye drops on macular blood flow density and retinal thickness in children with different degrees of myopia. METHODS: This was a prospective case-control study. Sixty-four patients (112 eyes) diagnosed with myopia for the first time with 0.01% atropine eye drops before and 6 months after medication were investigated with the uncorrected distance visual acuity (UCVA), axial length (AL), spherical equivalent (SE), macular ganglion cell-inner plexiform layer thicknes (mGCIPL) using slit lamp examination and optical coherence tomography (OCT), vascular density in the macular area and the area of the avascular in the fovea using optical coherence tomography angiography (OCTA) . Changes in various indicators before and after medication were compared. RESULTS: Compared with before medication, the AL of the three groups of myopia patients increased significantly (P0.05). The difference was statistically significant between the moderate myopia group and the high myopia group (P0.05). After 6 months of medication, the central circle macular vessel density (cCVD) increased in the low myopia group and moderate myopia group (P0.05). Before and after medication, there was no significant difference in outer circle macular vessel density (oCVD), inner circle macular vessel density (iCVD), and whole circle macular vessel density (wCVD) among the three myopia groups (P>0.05). The increase in mGCIPL was statistically significant in the low myopia group (P0.05). There was no significant difference in foveal avascular zone (FAZ) among the three myopia groups before and after medication (P>0.05). There was no correlation between CVD, AL, and SE in the three myopia groups (P>0.01). There was a low correlation between CVD and mGCIPL in the low myopia group (r=0.442, P0.01). CONCLUSION: 0.01% atropine can significantly reduce the rate of axial and refractive growth in children with low to moderate myopia, increase the density of central macular vessels, and increase the thickness of mGCIPL in children with low to moderate myopia.
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The purpose of this paper is to review the research progress on the effects of obstructive sleep apnea （OSA） on memory consolidation， and to speculate on possible mechanisms underlying these effects， so as to inform the exploration of effective therapeutic measures for impaired memory consolidation. Previous studies have shown that mild OSA may impair different types of memory consolidation， and the impairments are closely related to certain indices of polysomnography （such as sleep microstructure， apnea hypopnea index， arousal index， etc）. Therefore， it is hypothesized that disruption of sleep architecture and damage to brain regions and neural pathways associated with sleep-dependent memory consolidation due to intermittent hypoxia may trigger a decline in memory consolidation. Meantime， long-term continuous positive airway pressure can alleviate the impairment of memory consolidation induced by OSA， but whether other interventions can mitigate the damage remains unclear.
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Objective To analyze the clinical efficacy of AngioJet mechanical thrombus aspiration system for patients with acute pulmonary embolism (PE).Methods Clinical data of 28 cases of acute pulmonary embolism (PE) patients was retrospectively analyzed,8 cases (AngioJet group) were treated with AngioJet + CDT,20 cases were treated by pigtail catheter thrombolysis(CDT group)alone,the total amount of urokinase,thrombolytic time,related detection index and the occurrence of complications were compared between the two groups.Results The dosage of urokinase in the two groups was (72.5 ± 44.4) × 104U and (169.0 ± 59.3) × 104 U respectively,P ＜ 0.05.The catheter indwelling time was (1.0 ± 0.89) days and (2.65 ± 0.86) days respectively (P ＜ 0.05).There were no statistically significant differences in SBP,PaO2,SpO2 and D-dimer between the two groups before and after operation (P ＞ 0.05).Conclusion Both AngioJet and CDT are effective methods for the treatment of acute PE.The combination of the two methods can accelerate the improvement of clinical symptoms,reducing the dosage of thrombolytic drugs and the occurrence of surgery-related complications.
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Objective To determine the potential deficiency due to lack of anaerobic culture and evaluate the effect to reduce adverse reaction associated to transfusion-translated bacterial infection.Methods The result of 9 758 units of apheresis platelet concentrates (PCs)detected with automated microbial detection system were reviewed and the medical records of the patients that received the contaminated PCs were followed.Results The confirmed positive rates by aerobic and anaerobic cultures were 0.06％ (6/9 758)and 0.16％ (16/9 758),respectively.In 10 of 16 yield cases,only the anaerobic culture was positive.The most of the bacterial detected by anaerobic culture only were Propionibacterium acnes.Their mean detection time from inoculation was 96.8±18.21 hours.Conclusion Addition of anaerobic culture would enhance the detection of bacterial contamination in PCs.However,since only slow-growing bacteria were detected,and because their clinical significance was debatable,blood service should select feasible and costeffective projects using only aerobic bottle for bacterial screening,like the majority of licensed blood centers in North America and Hong Kong,China.
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Objective To evaluate clinical outcome of patients treated with endovascular dilation and stent placement for ilio-femoral venous postthrombotic syndrome (PTS).Methods 42 PTS cases were enrolled in our group from June 2003 to December 2011.Dilation and stent placement in stenosis/ occlusion of ilio-femoral veins were performed in all cases,temporary femoral arteriovenous fistula was established in 24 cases.Results These were not severe perioperative complications.All patients were followed up for 8-75 months.Preoperative limb ulcers in 5 cases were healed.Early thrombosis in stents found in 3 cases was cleared by catheter directed thrombolysis (CDT).Severe in-stent restenosis (＞ 50％)was found in 10 cases,treated with dialation or re-stenting.Primary patency,assisted primary patency and secondary patency rates were 75％,78％,80.1％;64.1％,67.1％,69.6％;63.8％,65.3％,66％;61.7％,65％,65.2％,respectively,at 6 months,12 months,24 months and 36 months.Postoperative Venous Clinical Severrity Score (VCSS) decreased (P =0.000).There was statistical significant improvement in every dimension evaluated by the MOS item short from health survey(SF-36) after operation (P =0.000).Conclusion Ilio-femoral venous stenting is a safe and effective treatment for PTS and with good clinical midterm results.
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Objective To investigate the inhibitory effects of histidine grafted poly (β-amino es-ter) ( HGPAEs) vector-based RNA interference ( RNAi) on the expression of gene encoding myeloid differ-entiation factor 88 (MyD88) in rat liver tissues.Methods The sequence of small hairpin RNA (shRNA) was designed based on the genetic information of MyD 88.HGPAEs vector was constructed and coupled with shRNA plasmid targeting MyD88 to construct pMyD88-HGPAEs vector.Rats were divided into five groups including control group , HGPAEs treatment group , pHK-HGPAEs treatment group , shRNA treatment group and pMyD88-HGPAEs treatment group .The rats in each group were transfected with the corresponding inter-ventions through portal vein injection .Real-time PCR and Western blot assay were performed to detect the expression of MyD88 in liver tissues 3 days after transfection .Results The pMyD88-HGPAEs vector was successfully constructed .The expression of gene encoding MyD 88 was inhibited in rats from shRNA treat-ment group and pMyD88-HGPAEs treatment group (P<0.05).Significantly decreased expression of gene encoding MyD88 at mRNA and protein levels were observed in rats from pMyD 88-HGPAEs treatment group as compared with those from other groups (P<0.01).Conclusion HGPAEs vector might be used as a po-tential gene carrier .The expression of gene encoding MyD 88 in rat liver tissues could be significantly inhibi-ted through portal vein injection of pMyD 88-HGPAEs vector .This study provided evidences for further re-search on pMyD88-HGPAEs vector in a high responder model of rat orthotopic liver transplantation .
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OBJECTIVE@#To investigate the effect of dxamethasone (DEX) on the expression of Tregs in allergic rhinitis (AR) mice, and explore the mechanism of glucocorticoid in the treatment of AR.@*METHOD@#AR murine model was established by sensitization and challenge with OVA, besides intervention treatment with DEX was carried out in AR model. The behavior observation was used to evaluate the improvement effect of DEX on AR symptoms. The morphological characteristics of nasal tissues were observed by HE staining after fixation and decalcification. The mononuclear cells were obtained by grinding spleens, and the total RNA was extracted for reverse transcriptase polymerase chain reaction to investigate the level of mRNA expression of Foxp3. The changes of CD4+ Foxp3+ Tcells in spleen of mice were analyzed by flow cytometry.@*RESULT@#BALB/c mice received OVA sensitization followed by OVA intranasal challenge, the frequencies of sneezing and nose-scratching increased significantly in AR group (44. 50 ± 5. 61 and 72. 94 ± 8. 76) compared with control group (12. 68 ± 1. 87 and 26. 76 ± .2. 89), P<0. 01; The frequencies decreased significantly in DEX group (26. 04 ± 3. 93 and 56. 79 ± 5. 64), P< 0. 05 compared with AR group. The continuity of nasal mucosa ciliated columnar epithelium in AR group was destroyed and appeared to be repaired in DEX group. Inflammatory cells infiltration was also markedly decreased by DEX treatment. The proportion of CD4+ Foxp3+ T cells in AR group (3. 89 ± 0. 39)% decreased, P<0. 01 vs control group (4. 63 ± 0. 15) %. DEX treatment induced production of Tregs (6. 89 ± 0. 49)%, P<0. 05 vs control group. DEX significantly increased the expression of Foxp3 mRNA (P<0. 05) compared with AR and control group.@*CONCLUSION@#DEX reduce upper airway allergic inflammation effectively, which may be mediated by promoting the expression of Foxp3 and inducing the amplification of Tregs in vivo.
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Animals , Mice , Administration, Intranasal , Dexamethasone , Pharmacology , Disease Models, Animal , Flow Cytometry , Forkhead Transcription Factors , Metabolism , Inflammation , Drug Therapy , Mice, Inbred BALB C , Nasal Mucosa , Ovalbumin , RNA, Messenger , Rhinitis, Allergic , Drug Therapy , T-Lymphocytes, RegulatoryABSTRACT
Objective To investigate the inhibitory effects of cationic polymeric liposomes (CPLs) vector-based RNA interference (RNAi) technology on the expression of rat MHCⅡ transactivator ( CⅡTA) and MHCⅡgenes .Methods According to the genetic information of CⅡTA downloaded from GenBank, three short hairpin RNA (shRNA) sequences targeting CⅡTA sequences were designed .CPLs vectors were constructed and coupled to shRNA plasmid vectors to form pCⅡTA-CPLs vectors .Six groups including control group , CPLs control group , pHK-CⅡTA control group and three pCⅡTA-CPLs groups were set up.Rat dendritic cells (DCs) were transfected in vitro.Real time PCR and flow cytometry analysis were used to detect the expression of CⅡTA and MHCⅡat mRNA and protein levels in DCs after transfec-tion.Results The pCⅡTA-CPLs vectors were successfully constructed .Compared with control groups ,the transcription level of CⅡTA and MHCⅡand the expression of MHCⅡat protein level were significantly in-hibited in all pCⅡ TA-CPLs groups ( P<0 .01 ) .The strongest inhibitory effects of pCⅡTA-CPLs on the ex-pression of CⅡTA and MHCⅡgenes were observed in the second pCⅡTA-CPLs group.There was a positive correlation between the expression of CⅡTA and MHCⅡ.Conclusion CPLs vectors were effective gene carriers.The constructed pCⅡTA-CPLs vectors significantly inhibited the in vitro expression of rat CⅡTA and MHCⅡ, which provided evidences for further investigation on pCPLs-CⅡTA vectors in vivo.
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Retrospective analyses of clinical data were performed for 518 elderly cataract patients (856 eyes) of Disabled Persons' Federation in Hangzhou,Zhejiang Province.For 69 eyes (8.06％),corneal edema was a major factor of low vision during early postoperative period; for 23 eyes (2.69％),delayed cataract was an important cause of low vision during long-term postoperative period.The best corrected visual acuity of 120 eyes caused by nonsurgical factors was diabetic retinopathy (48 eyes,48.00％),senile macular degeneration (25 eyes,20.80％),high myopia retinopathy (17 eyes,14.20％),optic never injury in glaucoma (16 eyes,13.30％),macular membrane & hole (5 eyes,4.20％) and amblyopia & corneal leucoma (9 eyes,7.50％).Retinal disease is the most important reason of low vision after cataract surgery.And surgery remains the most effective treatment for cataract.
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Objective To explore the effects of CD4+CD25+Treg cell on the allograft after infusion of dendritic cells (DCs) with low expression of CD40 from donor in mouse heart transplantation.Methods In vitro,mouse bone marrow-derived DCs were infected by CD40-RNAi lentiviral vector,and tolerogenic DCs (Tol-DCs) with low expression of CD40 were prepared.A heterotopic abdominal heart transplantation model was established in mice,and the other three groups that were control group,noninfected DC group and lentivirus infected DC group were designed correspondingly.Cardiac allograft survival time was recorded and pathological grading for acute rejection was assessed on the 7 d after heterotopic abdominal heart transplantation.Concentrations of CD4+CD25+Treg cells in peripheral blood were analyzed before and after transplantation by flow cytometry.Results After 48 h infection of DCs by CD40-RNAi lentiviral vector in vitro,the expression of CD40 mRNA was down-regulated significantly,whose inhibition rate was 80.9％.The expression of CD40 was decreased from 74.37％ ±4.08％ to 40.07％ ± 4.03％ (P＜0.05) after 48 h infection.Compared with the control group and the noninfected DC group,the cardiac allograft survival time was significantly prolonged in the CD40-RNAi lentivirus infected DC group,which was (14 ± 4) d(P＜0.01) ; concentrations of CD4+CD25+Treg cells in peripheral blood were increased both on the 3 d and the 7 d after transplantation (P＜0.05) ; the pathological grading for acute rejection was decreased on the 7 d after transplantation (P＜0.05).Conclusion The CD4+CD25+Treg cell in peripheral blood was protective to cardiac allograft in prolonging its survival time in mouse heart transplantation.
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Objective To evaluate the safety and efficacy of SilverHawk directional atherectomy device in the treatment of arterial stenoses/occlusions of the femoropopliteal regions.Methods From April 2011 to May 2012,36 patients (28 men,8 women,age range 60 - 84y) with 40 arteriosclerotic lesions of femoro-popliteal arteries (de novo lesions in 25 arteries,in-stent restenosis in 15 arteries; Rutherford score of 3 - 5 ) were included in the treatment by SilverHawk directional atherectomy device.All the patients were diagnosed via low-extremity artery CTA and arteriography.The mean ankle brachial index (ABI) of treated limbs was 0.53 ±0.12.Results The overall technical success rate was 100％ (40/40).The procedural success rate was 87.5％ (35/40).There was no perioperative mortality with three cases developing complications,vascular injury occurred in two cases.One was treated by stent-graft while the other was managed conservatively; Procedure-related arterial embolization occurred in one case and was treated by suction through catheter.Symptoms were relieved in all patients,ABI increased to 0.72 ± 0.18.All the patients were followed up for an average of (6.4 ± 1.2) months.Postoperative restenosis developed in one case and treated with balloon angioplasty and stent placement after six month.Conclusions SilverHawk directional atherectomy device is effective and safe in treament of arterial stenoses/occlusions of the femoropopliteal lesions,with satisfactory early results.
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Objective To investigate the clinical correlative factors of malignant cystic pancreatic tumors.Methods The clinical data of 45 patients who received cystic pancreatic tumor resection at the General Hospital of Tianjin Medical University from May 2000 to May 2011 were retrospectively analyzed.All patients were divided into benign tumors + precancerous lesions group (35 patients) and malignant tumor group (10 patients).The clinical symptoms and imaging features of cystic pancreatic tumors were analyzed.All data were analyzed by chisquare test or Logistic regression analysis.Results Abdominal pain,jaundice,emaciation,nausea and vomiting were observed in 23 patients (51％),and 22 (49％) patients had no clinical symptoms.The clinical features of benign pancreatic cyst included pancreatic calcification and pancreatic divisum,while the clinical features of malignant pancreatic cystic tumors were nodules,swelling of lymph nodes,dilation of biliary and pancreatic duct.The results of univariate analysis showed that age ≥ 60 years,presence of symptoms,jaundice,emaciation,dilation of pancreatic duct were the correlative factors of malignant cystic pancreatic tumors ( x2 =4.220,4.294,4.645,7.705,4.645,P ＜ 0.05 ).The results of multiple logistic regression analysis found that age ≥60 years,dilation of pancreatic duct and presence of clinical symptoms were the correlative factors of malignant cystic pancreatic tumors ( OR =1.573,2.674,2.723,P ＜ 0.05).Conclusion Age≥60 years,dilation of pancreatic duct and presence of clinical symptoms are the correlative factors of malignant cystic pancreatic tumors.
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Objective To evaluate balloon angioplasty and stenting of the hepatic veins for the treatment of Budd-Chiari syndrome with long segmental inferior vena cava (IVC).Methods The hepatic veins were evaluated by colour Doppler,CT or MR.Epidiaphragmatic inferior vena cavography was performed to locate the stenused opening of the hepatic veins,in case of membrane occlusion of the hepatic veins,a puncture was attempted with Rups100 needle and then balloon dilation of the hepatic veins and stents implantation was carried out.Results In 40 cases of long segmental inferior vena cava ( IVC ) occlusive Budd-Chiari syndrome,membranous obstruction of the hepatic veins was found in 5 cases and short-segmental occlusion in 24 cases.28 cases were successfully treated with balloon dilation and stents implantation,including percutaneous transluminal angioplasty(PTA) of hepatic vein in 5 cases,and stent implantation in 23 cases,puncture procedure was failed in 1 case.Postoperative follow-up was made in 26 cases from 6 to 62 months ( mean,24.0 ± 1.3 months ).Symptoms recurred in 6 cases.Hepatic vein restenosis or occlusion were observed using color Doppler ultrasound in 6 cases.Conclusions Most lesions in hepatic veins were membranous obstruction or short-segmental occlusion among patients with long-segmental occlusion of IVC.Balloon angioplasty and stenting of hepatic veins for long segmental IVC occlusive Budd-Chiari syndrome through jugular vein can relieve hepatic venous obstruction and relieve portal hypertension.
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Objective To summarize the experience on treatment for 15 cases of acute pulmonary embolism(PE).Methods Fifteen acute PE patients admitted from June 2009 to May 2011 were analyzed retrospectively.All patients were diagnosed as PE and deep vein thrombosis,and treated with placement of inferior vena caval filters(IVC).Five patients with main pulmonary artery embolism accepted intrapulmonary arterial interventional therapy of thrombus fragmenlation and suction and catheter-directed thrombolysis (CDT).Ten patients with embolization on pulmonary artery branch and acute iliofemoral vein thrombus accepted therapy of peripheral thrombolysis.During postoperative course improvement was observed on the clinical symptoms,occurrence of complications,Miller index,change of mean pulmonary arterial pressure (mPAP) and arterial partial pressure of oxygen(PO2),as well as the patency of pulmonary artery.Result Five main pulmonary artery embolization patients gained complete patency of pulmonsnary artery,and the clinical symptoms immediately improved.Miller index reduced from (0.51 ± 0.04) to (0.27 ± 0.38),mPAP decreased from (55.3 ± 3.1 ) mm Hg to ( 32.7 ± 2.2 ) mm Hg,and PO2 elevated from ( 40 ±3 ) mm Hg to ( 63 ± 4) mm Hg,showing a significant difference ( P ＜ 0.01 ).Ten patients with pulmonary artery branch embolization gained patency of pulmonary artery branch,iliofemoral venous thrombosis cleared,and clinical symptoms significantly improved.All patients recovered after two weeks of intravenous thrombolytic,anticoagulation and antiplatelet therapy.During three to twelve months' follow up,the therapeutic effects persisted and there was no recurrence.Conclusions Emergency intrapulmonary arterial interventional therapy of acute PE has remarkable effectiveness,safety and feasibility,improving pulmonary obstruction and clinical symptoms.
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ObjectiveTo investigate the feasibility and effect of complete thoacoscopes and laparoscopes combination operation for thoracoabdominal injuries.MethodsFrom March 2007 to March 2011,the clinical data of 36 patients (observation group) with thoracoabdominal injuries who were performed with complete thoracoscopes and laparoscopes combination operation were analyzed retrospectively,and compared with 36 patients (control group) treated with traditional standard operation in the same period.The intraoperative and postoperative situation was compared between two groups.ResultsAll the patients in observation group were successfully performed by complete endoscopic surgery.There were no operative mortality and complications related to operation,such as diaphragmatic hernia,delayed hemopneumothorax.The operative time of observation group was (98 ±38 ) min,operative blood loss was ( 120±45 ) ml,drainage flow within 24 h was(230 ±55) ml,drainage tube duration was(5 ±2) d,postoperative pain vasual analogue scale (VAS) score was (3.31±0.87) scores,hospitalization cost was ( 1 2.3±7.6) thousand yuan,postoperative recovery time of intestinal function was (2.29±1.02) d,hospital stay was ( 10±3 ) d.Those values of control group were respectively ( 135±31 ) min,(220±30) ml,(400±160) ml,(9±3) d,(6.82±1.67) scores,(23.4±8.5) thousand yuan,(5.46 ±2.31 ) d and ( 16±2) d.There were significant differences between two groups(P ＜0.05).The occurrence rate of cardiopulmonary complication of observation group [ 11.1％(4/36) ] and control group [ 13.9％(5/36) ] had no significant difference (P＞0.05).ConclusionsComplete thoracoscopes and laparoscopes combination operation for thoracoabdominal injuries is a safe and mini-invasive treatment with quicker recovery,less cost and morereliable effect.Therefore,it deserves further clinical application.
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Objective To summarize the therapeutic experience on malignancy patients complicated with hyperthyroidism.Methods Clinical date of 10 cases of surgical malignancy complicated with hyperthyroidism admitted from May 2004 to May 2010 in Tianjin Medical University General Hospital were analyzed retrospectively.Results Four cases were treated by subtotal thyroidectomy before radical operation for cancer.Radical operation for cancer was performed on 6 patients after clinical symptoms of hyperthyroidism were controlled by perioperative antithyroid agents.Postoperatively 3 patients complicated with clinical manifestations similar to thyroid crisis.There was no postoperative mortality.Chemotherapy was given to 10 patients,and 1 patient was discontinued for chemotherapy caused leucopenia.Conclusions Hyperthyroidism should be controlled by surgery or antithyroid agents before patients of malignant diseases could proceed with radical surgery.Proper preoperative medication and effective postoperative management can reduce operation risk and help the patients get through the perioperative period safely.
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Objective To evaluate a combination of interventional treatment and surgical exploration for acute lower limb ischemic disease.Methods We reviewed 42 cases admitted from July 2007 to January 2010,all patients complained pain,paralysis,pulselessness,pallor and paresthesia.After Fogarty thrombectomy angiography was taken in DSA room.Patients with angiostenosis greater than 50％ were then managed by interventional treatment(CDT,PTA,Stenting).Results Lives were saved in all patients,40 lower limbs were saved,and 2 patients received below knee amputation.The amputation rate was 4.76％.Dorsal or(and)posterior tibial artery of foot was felt in 33 patients,symptoms significantly improved.The other 7 patients still had painful and paralysis on the diseased limb.Conclusions The interventional treatment and surgical operation in acute lower limb ischemic disease is safe and result is satisfactory,which can improve the long-term patency and salvage rate of the lower limb.
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Objective To summarize the experience of treating the end stage of liver disease complicated with renal failure using combined liver-kidney transplantation.Methods The clinical data of 28 cases receiving combined liver-kidney transplantation were retrospectively analyzed, including the inclusion criteria of surgical indications, modus operandi, protocol of immunosuppression and the prognosis post-operation.Results Among these 28 cases in our study, 22 cases suffered from liver and renal failure, accounting for 78.6%; 4 cases were diagnosed as having hepatorenal syndrome, accounting for 14.3%; and 1 case had hyperoxaluria and polycystic liver with polycystic kidney. As for the modus operandi we used, piggy-back procedure was adopted for 4 patients and classic procedure without bypass was used for the rest. Donor kidneys were all put in the right iliac fossa. During the follow-up period of 5 months to 7 years, one-and 3-year survival rate of the recipients was 92.9% and 78.3% respectively. Among these 28 recipients, 4 cases had the graft renal dysfunction early post-operation: One died and 3 recovered through consecutive therapy. One case received re-transplantation of the liver 3 months after the first due to the relevant complications and then recovered. During this period, no impact on the renal function occurred. Eleven cases had pulmonary infection post-operation, and 1 died. No acute rejection occurred.Conclusion Combined liver-kidney transplantation is the effective treatment to the patients with end stage liver disease complicated with renal dysfunction. Suitable case selection and perfect operation timing were the key points to the success of combined liver-kidney transplantation.
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Objective To investigate the effect of blocking CD40/CD40L costimulatory pathway by the lentiviral vector-mediated RNA interference on the survival of mouse cardiac allograft. Methods Mouse bone marrow-derived dendritic cells (DCs) were infected by CD40-RNAi lentiviral vector in vitro, and tolerogenic DCs (Tol-DCs) with decreased CD40 expression were prepared. Fluorescence real-time quantitative PCR and flow cytometry were used to analyze the expression of CD40 mRNA and DC surface antigens CD40, CD11c, MHC Ⅱ before and after infection. Mouse model of heterotropic abdominal heart transplantation was established. Seven days prior to heart transplantation, Tol-DCs with decreased CD40 expression were transfused into recipient mice intravenously (lentivirus infected DC group). Control group and non-infected DC group were assigned simultaneously. The survival of cardiac allograft was monitored and pathological grade of acute rejection 7 days after heterotropic abdominal heart transplantation was determined. Results The transcription of CD40 mRNA of DCs was down-regulated significantly at 48 h after CD40-RNAi lentiviral vector infection, and the inhibition rate was 80. 9%. The expression of CD40 protein was also significantly decreased as compared with control group (40. 07% ± 4. 03% ) ( P ＜ 0. 05 ).Compared to control group (8 ± 2 days) and non-infected DC group (9 ± 1 days), the survival time of cardiac allograft in CD40-RNAi lentivirus infected DC group (14 ± 4 days) was significantly prolonged (P＜ 0. 05 ), and the pathological grade of acute rejection decreased significantly ( P ＜ 0. 05 ).Conclusion Blocking CD40/CD40L costimulatory pathway could hamper the activation of allogeneic T lymphocyte, inhibit the acute rejection and prolong the survival of mouse cardiac allograft.
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Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.