Objective To discuss the way and management of extracorporeal circulation (ECC) for totally robotic assisted cardiac surgery. Methods A Total of 226 patients underwent cardiac surgery using da Vinci S robotic surgical system, including 111 patients underwent atrial septal defect repair ( ASDR) , 9 patients underwent ventricular septal defect repair ( VSDR) ,51 patients underwent mitral valvuloplasty ( MVP) , 20 patients underwent mitral valve replacement( MVR) , 27 patients underwent left atrial myxoma excise and 4 patient underwent right atrial myxoma excise. ECC for most of patients was achieved with femoral arterial cannula, femoral venous cannula and right internal jugular venous cannula, except for 1 patient underwent MVP with femoral arterial cannula and femoral two-stage venous cannula. In all the cases, vacuum-assist venous drainage ( VAVD) , continuous blood gas monitoring and ultrafiltration were used during ECC. myocardial protection was pertic cross-clamp time was 40 ～219 (105.9+38. 8)min and 21 ～166 (69.5±30.0)min respectively. During ECC, the mean urine volume was 100-2100 (771.7±477.6) ml, ultrafiltration volume was 1000-4800 (2495.4 ±811.6) ml, and the total fluid balance was subzero-balanced (172 cases) or zero-balanced(13 cases) in most of patients. Conclusion The establishment of ECC system through peripheral vessels, using VAVD and continuous blood gas monitoring are the key points of ECC for totally robotic assisted cardiac surgery, also certain learning curve of perfusion technique and close communication between the surgical team are essential during ECC.

Objective Todetecttheexpressionlevelsoftumornecrosisfactor-α(TNF-α)and interleukin-6(IL-6)inintracranialaneurysms.Methods Sixteenconsecutivepatients(aneurysm group)with intracranial aneurysm confirmed by digital subtraction angiography (DSA)and clipped by microneurosurgery were enrolled retrospectively. A total of 19 trauma patients without vascular disease confirmed by CT and magnetic resonance imaging (MRI)in the same period were used as a control group. Hematoxylin-eosin (HE)staining and immunohistochemical staining were used to detect the aneurysm wall tissue and the colored portions of TNF-α and IL-6 in normal vessel wall,the mean value of optical density after its expression was analyzed,and the intensity of staining was compared. Results (1)Each layer of artery walls of the control group had no obvious TNF-α and IL-6 expression. The inner,media and out membranes of the aneurysm wall tissue of the aneurysm group had positive expression of TNF-αand IL-6. (2)The mean optical densities of TNF-α and IL-6 in patients of the aneurysm group were 0. 182 ± 0. 069 and 0. 148 ± 0.062 respectively,and they were higher than 0. 144 ± 0. 031 and 0. 105 ± 0. 020 of the control group. The differences were statistically significant (all P<0. 05). (3)The mean optical densities of TNF-α expression of each layer of the inner,media and out membranes in the aneurysm walls were 0. 224 ± 0. 071,0. 134 ± 0. 040,and 0. 106 ± 0. 065,respectively. There were significant differences (P<0.01). (4)The mean optical density expressed by IL-6 in the out membrane of the aneurysm walls was lower than the media and inner membranes (0. 096 ± 0. 018 vs. 0. 145 ± 0. 050,and 0. 148 ± 0. 070). There were significant differences (P<0. 05). (5)The results of Spearman correlation analysis showed that the mean optical density of TNF-αof the aneurysm group was positively correlated with that of IL-6 (r=0. 452, P<0.05).Conclusion TheexpressionlevelsofTNF-αandIL-6intheaneurysmwalltissueare higher,and they may be involved in intracranial aneurysm formation and rupture.

Objective To study the safety and efficacy of low dose Peginterferon-alpha-2a (PEG-INF-α-2a) combined with Ribavirin treating chronic hepatitis C in renal transplant recipients.Methods A total of 13 cases of HCV hepatitis were randomly divided into treatment and control groups.Seven cases in treatment group were given PEG-INF-α-2a (90 μg/week) and ribavirin (600mg/day) for 16 to 48 weeks,and the rest 6 cases in control group were subjected to general liver protection and anti-inflammatory treatment. All patients were followed up for more than 2 years.Results There were 5 cases getting early response in treatment group for 16 weeks,including four cases of complete response and no non-effects response patients. In 4 cases voluntarily receiving treatment for 48 weeks,1 case had facial muscle myalgia and increased Cr level at 35th week,humoral graft rejection was confirmed pathologically,and the treatment was terminated; 1 case had recurrence of HCV RNA replication and PEG-INF-α-2a was withdrawn at 38th week.As results,5 patients in the treatment group obtained complete response after two years,including 2 cases whose HCV-IgG had got negative,HCV RNA replication was significantly lower than in the control group,and the average Cr higher than in control group (P＞ 0.05). There were adverse reactions during this treatment protocol: fever,muscle myalgia,agranulocytosis, anemia and humoral graft rejection.Conclusion The efficacy of low lose PEG-INF α-2a combined with ribavirin is definite in the treatment of chronic HCV hepatitis in kidney transplant recipients.The 16-week treatment duration is reasonable.It is remarkable that PEG-INF-α-2a may cause humoral graft rejection and Cr crawling.

Objective To investigate the compensatory changes in morphology,function,and hemodynamic indices of the retained kidney after nephrectomy among living related donors.Method The 170 living related kidney donors underwent assessments before surgery as well as at 1st and 12th month,postoperatively,including length,width,short diameter,glomerular filtration rate (GFR),effective renal plasma flow (ERPF),peak systolic blood flow velocity (Vsmax),resistance index (RI),as well as pulsatility indices (PI) of main renal artery (MRA),segmental renal artery (SRA),and interlobar renal artery (IRA).Results All subjects were followed up for 9 to 68 months,with no observed hypertension or kidney failure.The length,width,and short diameters of the retained kidney were increased significantly (P＜0.01) at 1st and 12th month postoperatively.The renal sizes at 1st month postoperation were similar to that at 12th month postoperation (P＞0.05).GFR and ERPF were increased significantly as compared with preoperative values (P＜0.01) with similar values at postoperative month 1 and 12 (P＞0.05).The Vsmax of MRA,SRA,and IRA in the retained kidney were increased significantly (P＜0.01),and the RI and PI were also increased as compared with the preoperative values (P＜0.05),albeit these indicators were similar at postoperative months 1 and 12 (P＞0.05).Conclusion For all subjects studied after unilateral nephrectomy in a living related donor,the diameter of the retained kidney as well as the GFR and ERPF showed compensatory increases.Various arterial hemodynamic parameters also showed compensatory changes.Under strict donor inclusion criteria,living related kidney donor procedures is safe.

Objective To investigate the effect of external fixator on repairing the vascular injuries in the knee and calf.Methods From May 2004 to October 2010,208 patients (234 sides) with vascular injuries in knee and calf,treated with external fixation,were retrospectively analyzed,including 192 males (217 sides) and 16 females (17 sides) with an average age of 39.6 years (range,14-67).Blood vessel damage:the popliteal artery injury in 84 patients (86 sides),the anterior tibial artery injury in 36 (46),the posterior tibial artery injury in 41(49),the anterior tibial artery and the posterior tibial artery injury in 47 (53).Complete ischemic of the injured distal limb existed in 129 patients(141 sides),incomplete ischemia in 79 patiets (93 sides).All patients underwent emergency surgery.The AO external fixators were used in 164 patients,the chuangsheng inlaid external fixators in 37,and the hybrid external fixators in 7.The injured vessels were anastomosed directly,or indirectly with great saphenous vein.The wounds were treated with phase Ⅰ bandage,skin grafts,flap transplantation or VSD suction.Results One hundred and ninty four patients obtained successful limb salvage; but 14 patients underwent amputation owing to serious damage of limb and poor general condition.One hundred and twenty seven patients underwent the second stage debridement combined with skin graft or flap transplantation; 24 patients underwent the first stage free skin graft or flap transplantation;wound cicatrized by oneself in 43 patients.One hundred and forty eight patients were followed up for 8 months to 3 years.External fixators were removed after an average of 7.2 months (range,1-18).The average healing time of fracture in 102 patients was 5.6 months (range,3-13).Forty patients suffered bone nonunion or bone defect; after treated by fracture fixation,bone graft or bone transport,all of them got bone union.Conclusion The external fixation can shorten operation time as well as the time of limb ischemia,which is an effective option on repairing vascular injuries in knee and calf.

AIM To study the flavonoids from the leaves of Astragalus membranaceus (Fish.) Bge..METHODS The ethyl acetate and n-butyl alcohol fractions of 75％ ethanol extract from A.membranaceus leaves were isolated and purified by silica,ODS and preparative HPLC column,then the structures of obtained compounds were identified by spectral data.RESULTS Thirteen compounds were isolated and identified as quercetin (1),quercetin-3-O-β-D-glucopyranoside (2),rhamnocitrin-3-O-β-D-glucopyranoside (3),rhamnocitrin-3-O-β-neohesperidoside (4),rhamnocitrin-3-O-3-D-glucopyranoside (1'''→2'')-β-D-apiofuranosyl (5),complanatuside (6),glycitein (7),4',7-dihydroxy-3-methoxy isoflavone (8),genistein (9),calycosin-7-O-β-D-glucopyranoside (10),genistin (11),glycitin (12),tiliroside (13).CONCLUSION Compounds 5,8,13 are isolated from genus Astragalus for the first time,and compound 2 is first isolated from this plant.

Objective To evaluate the efficacy of benvitimod on allergic contact dermatitis (ACD) in a mouse model of allergic contact dermatitis.Methods Acute and chronic ACD models were established respectively in 42 BALB/c mice through 2,4-dinitrofluorobenzene (DNFB) sensitization and challenge.Then,the BALB/c mice were equally divided into 7 groups with 6 mice in every group:normal control group receiving no treatment,five treatment groups topically treated with 0.1％ dexamethasone solution,0.1％ tacrolimus (FK506) solution,0.5％ benvitimod solution,1.0％ benvitimod solution and 2.0 ％ benvitimod solution respectively,and dehydrated alcohol group treated with dehydrated alcohol only.All the drug solutions were topically applied at 24 and 36 hours after the last challenge in the murine models of acute ACD which were sacrificed at 48 hours,and twice daily from day 7 to 21 after the initial sensitization in the murine models of chronic ACD which were sacrificed on day 21 after the first topical treatment.Ear tissues were obtained from these mice and subjected to measurement of ear thickness and weight,as well as pathological examination and evaluation of inflammatory infiltrate by hematoxylin-eosin (HE) staining.The safety of these drugs was also estimated at the end of treatment.Results In the murine models of acute ACD,benvitimod showed no obvious therapeutic effect at 24 hours,with no significant differences in bilateral difference in ear thickness or weight between the three benvitimod groups and dehydrated alcohol group (all P ＞ 0.05).Meanwhile,in the murine models of chronic ACD,benvitimod markedly decreased the swelling degree of ears,with significant differences between the three benvitimod groups (0.5％,1.0％ and 2.0％) and dehydrated alcohol group in bilateral difference in ear thickness ((71.50 ± 3.15) × 10-3 mm,(75.50 ± 3.02) × 10-3 mm and (69.50 ± 2.59) × 10-3 mm vs.(91.83 ± 2.04) × 10-3 mm,all P＜ 0.01) and weight ((2.33 ± 0.45) mg,(2.30 ± 0.57) mg and (2.38 ± 0.27) mg vs.(3.73 ± 0.33) mg,all P ＜ 0.01) after 3 weeks of treatment.The inflammatory infiltration in ear tissue was significantly attenuated in murine models of both acute and chronic ACD by the three concentrations of benvitimod compared with dehydrated alcohol (all P ＜ 0.01).Conclusions Topical benvitimod can inhibit chronic ACD in mice induced by 2,4-DNFB,but exhibits no obvious effect on acute ACD.No apparent local adverse effects were observed during the treatment with benvitimod in these mice.

Abstract: Objective To construct a shuttle vector pHT315-AaCPR100A with two spore-producing-dependent promoters and the target gene AaCPR100A in Escherichia coli-Bacillus thuringiensis. Methods The forward promoter of Cry3A, named Pro-1 (+), was amplified by PCR using pSVP27A plasmid as the template, and the target gene AaCPR100A was amplified using Aedes aegypti RNA reverse conversion cDNA as the template. The plasmid pHT315 was linearized by digestion with Hind Ⅲ and Sal Ⅰ. The forward promoter and the target gene were inserted into the linearized vector pHT315 successively by in-fusion cloning according to the transcription direction. The synthesized plasmid containing the Cry3A reverse promoter sequence was used as the template, and the Pro-1 (-) reverse promoter was amplified by PCR. The intermediate vector containing the forward promoter and the target gene was linearized by EcoR I restriction enzyme, and the reverse promoter was inserted downstream of the target gene by in-fusion cloning in the direction of transcription. Results By agarose gel electrophoresis, the forward promoter, target gene AaCPR100A and reverse promoter bands were clear and of good quality, which could be used for in-fusion cloning experiments. The two spore-producing-dependent promoters and target gene fragments were connected by In-fusion cloning. The recombinant vector pHT315-AaCPR100A was verified by PCR. The forward promoter, target gene fragment and reverse promoter were successfully amplified in the recombinant vector. Nucleotide sequencing verified that the sequencing results of the bidirectional promoter sequence and the target gene sequence were basically consistent with the sequence alignment results, which met the requirements of the construction of vector elements and proved that the recombinant vector was successfully constructed. Conclusions Based on the above results, this study proves that the recombinant shuttle vector with two spore-producing-dependent promoters can be successfully constructed by in-fusion cloning technology, laying the foundation for the construction of engineered Bacillus thuringiensis expressing dsRNA of AaCPR100A.

Objective:To identify the hub differentially expressed genes(DEGs)of glomerular pathological changes and potential pathways in molecular process of type 2 diabetic nephropathy(DN)based on bioinformatics technology.Methods:The differentially expressed genes of Gene Expression Omnibus(GEO)dataset GSE96804 in DN and normal kidney tissues were analyzed by R 3.6.2 software. DEGs were further assessed by Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway analysis. Subsequently, the hub genes and their associated pathways were analyzed using String 11.0 and Cytoscape 3.7.2 software.Results:A total of 168 DEGs were obtained in the dataset. Among them, seven hub genes were identified, including ALB, FN1, EGF, PTGS2, PLG, KDR, and LOX. Three hub genes, ALB, EGF, PLG, exerted a direct action on glomerulus. GO enrichment analysis of DEGs was mainly manifested in extracellular matrix organization, extracellular structure organization, platelet degranulation and other biological processes, extracellular matrix, secretory granule lumen, platelet alpha granule and other cell components, chaperone binding, copper ion binding, antioxidant activity, and other molecular functions. DEGs mainly regulated metabolic process, which was related to fatty acid degradation signal pathway, exogenous substance metabolism related to CYP enzyme and drug metabolism signal pathway.Conclusion:A bioinformatics analysis of DN from the perspective of glomerulopathy is helpful to understand the potential molecular mechanism of DN and provide reference for further validation.

BACKGROUND:Chronic al ograft nephropathy is a complication of kidney transplantation and most of patients wil eventual y develop transplant kidney dysfunction. Bone marrow mesenchymal stem cells as a low immunogenicity special cellpopulation have been shown to have differentiation, transdifferentiation, paracrine and other basic functions, which have been successful used in other clinical areas. Based on this characteristic, bone marrow mesenchymal stem cells may play a therapeutic role in chronic al ograft nephropathy. OBJECTIVE:To study the safety and feasibility of autologus bone marrow mesenchymal stem cells transplantation via renal artery infusion and subsequent intravenous infusion guided by the digital subtraction angiography in the treatment of chronic al ograft nephropathy. METHODS:Eleven patients with chronic al ograft nephropathy who were confirmed from March 2011 to January 2013 were enrol ed, and then received transplant renal artery infusion once guided by the digital subtraction angiography and subsequent intravenous infusion twice of bone marrow mesechymal stem cells. Changes in serum creatinine, blood urea nitrogen, creatinine clearance, cystatin C, 24-hour urine protein, andβ2 microglobulin in the blood and urinary were monitored in patients up to 1 year after treatment. RESULTS AND CONCLUSION:Bleeding, transplant renal artery embolization, pseudoaneurysm and other related complications were not found in any of the 11 patients. The levels of serum creatinine, blood urea nitrogen and cystatin C were significantly decreased at 1 week and 1 month after celltherapy (P<0.05), while after 3 months of treatment, there was no difference before and after treatment (P>0.05). The creatinine clearance at 1 week and 1 month after treatment showed a remarkable increase, which were significantly different from that before treatment (P<0.05), but after 3 months of treatment, the difference was not significant (P>0.05). The level of 24-hour urine protein was significantly decreased after 7 days of treatment (P<0.05), and no difference was found after 1 month (P>0.05). The level ofβ2 microglobulin in the blood and urinary had no changes before and after treatment. These findings indicate that guided by the digital subtraction angiography, bone marrow mesenchymal stem cells via the renal artery infusion and subsequent intravenous infusion can improve kidney function of patients, but the celldosage and infusion method remain to be solved.