Objective To investigate the effects of medullary reaming on respiratory and circulatory system in dog model during total hip replacement. Methods Twelve mongrels were randomly divided into two groups. In control group, femoral necks were cut while femoral canals were not disturbed. In experimental group, femoral canals were reamed with femoral rasps as done in total hip replacement. The changes of hemodynamics and pulmonary function were monitored during perioperative period. The postmortem pulmonary tissues were studied by pathological examination. Results No fat embolus was found in any lung sections of control group. Meanwhile, pulmonary fat emboli were found in all of experimental group. After fat embolism happened, cardiopulmonary effects included increased pulmonary artery pressure, pulmonary shunt and pulmonary vascular resistance accompanying a decrease in partial pressure of oxygen and cardiac output. There is statistically significant difference in response between the two groups. Conclusion The present study showed that embolic events and intraoperative pulmonary impairment are common during reaming of femoral medullary canal in total hip replacements. Further research can be made with this model in order to reduce the risk of fat embolism syndrome.

Aim To study the screening of the nucleo-tide sequences might be affected by α-syn in vitro. Methods The nucleotide sequences were synthesized according to the feature of base composition, and then mixed with the α-syn-GFP. The CD was used to ana-lyse the changes of the peak. Result The peak of the CD changed greatly when the α-syn-GFP mixed with the GC-box like sequence. Conclusion The α-syn-GFP might affect the GC-box like sequence after trans-located into the nuclei. Then, it plays a role in physio-logical and pathological conditions by affecting the reg-ulation of gene expression.

Objective:To evaluate the effect of preoperative panel reactive antibody(PRA)levels on long-term survival after kidney transplantation. Methods:Data on 1 162 patients underwent first kidney transplantation performed between January 2001 and June 2014 were included in our center. According to the preoperative PRA levels,the patients were divided into negative group( PRA≤10%) and positive group( PRA>10%) ,which were retrospectively analyzed. Results: The 1-,5-,10-year patient survival rates of the negative group calculated by Kaplan-Meier were 96. 8%,89. 4%,78. 6%,respectively,while the positive group were 93. 5%,81. 6%, 65. 4%. The 1-,5-,10 -year death-censored graft survival rates of the negative group were 95. 9%,84. 8%,63. 1%,respectively,while the positive group were 92. 3%,74. 1%,51. 9%. The log-rank test revealed that there was significant difference between the patient and graft survival curves (χ2 =9. 623/11. 019, P=0. 002/0. 001 ) . Cox multivariate analysis found that preoperative PRA levels were independent risk factors for reducing the patient or graft survival rates(P<0. 001). Logistic multivariate regression analysis confirmed the significant association between preoperative PRA levels and the risk of acute rejection ( OR=8. 25,95% CI=2. 86-5. 72, P<0. 001). The 5-,10-year creatinine values were significantly lower in the negative group compared to the positive group(all P<0. 05), while there was no difference in the 1-year. In addition, Logistic multivariate regression analysis confirmed the significant association between preoperative PRA levels and the production of donor specific antibody(DSA)(OR=6. 89,95% CI=4. 52-9. 17,P<0. 05). Conclusion: The detection of preoperative PRA is an important indictor predicting the sensitivity status of the recipients. The preoperative PRA positive recipients need careful monitoring and diagnosis of acute rejection and DSA after kidney transplantation.

Aim To study of the expression and distribution of four α-synuclein truncations in three cells.Method Four α-synuclein gene truncations were obtained by PCR method,followed by subcloning into the pEGFP-N1 eukaryotic expression vector.Four obtained recombination plasmids were transfected into MN9D cells,PC12 cells and SH-SY5Y cells using Lipofectamine 2000 respectively.The expression and distribution of four α-synuclein truncations were observed by Confocal.Results Distribution of four α-synuclein truncations was discrepant obviously,the truncations,with more C terminal remained,were prone to emerging in nuclei.Conclusion Localization of α-synuclein protein in cells may be related to the C terminal,and the whole C terminal plays an important role in distribution of α-synuclein into nuclei.

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.

Objective To study the pathological changes of renal grafts from elderly donor in young recipients and to investigate the safety of kidney transplantation from elderly donors.Methods Fourteen elderly kidney donors (with the age >55 years old)and fourteen young recipients (with the age <30 years old)underwent living related donor renal transplantation at the Department of Transplantation of the Second Affiliated Hospital of Guangzhou Medical University from January 2008 to December 2008 were enrolled as the object of study.Every elderly donor kidney was performed time-zero biopsy and every young recipient was performed routine renal graft biopsy 6 months after transplantation.The pathological and structural changes of kidney tissues after renal transplantation from elderly donors were observed.Results The histopathological changes occurred at 6 months after transplantation from elderly kidney donors to young recipients.Compared with those before transplantation,the severity of renal arteriolar lesion and the index of renal arteriolar sclerosis alleviated after transplantation (P <0.05 ), and the ratio of glomerulosclerosis did not change after transplantation (P >0.05 ).The expression of fibronectin (FN)decreased after transplantation,but the difference had no statistical significance (P >0.05 ).The expression of laminin (LN ) decreased after transplantation (P <0.05).Conclusions The histopathological structure of renal graft from elderly donors in young recipients has improved.

Objective To investigate the application value of ImmuKnow immune cell function assay in monitoring of immune function changes after renal transplantation.Methods One hundred and six patients with uraemia undergoing renal transplantation in the Department of Organ Transplantation of the Second Affiliated Hospital of Guangzhou Medical University from January 201 3 to December 201 4 were included.Blood specimens were collected before transplantation and at the occurrence of infection or acute rejection during 1 2 months after transplantation.ImmoKnow was used to determine the adenosine triphosphate (ATP)content in CD4 +T cells.The ATP content of patients with renal transplantation at different clinical conditions were observed and compared,including periopreative group,stable group,acute rejection group and infecticn group (including severe pneumonia).The ratio of T cell subsets (CD4 +T cells,CD8 +T cells)and natural killer (NK)cells in peripheral blood were detected.Pearson correlation analysis was used to detect the association between ATP and the blood trough concentration of tacrolimus (FK506)and ciclosporin (CsA).Results The ATP content of the patients in the infection group was lower than that of the patients in the stable group (P <0.001 ).The ATP content of patients with severe pneumonia was lower than that of patients with other infections (P <0.05).The percentage of CD4 +T cells of the patients in the infection group was lower than that of the patients in the postoperative stable group (P <0.05 ). The ATP content was not associated with the postoperative blood trough concentration of FK506 and CsA.Conclusions ImmuKnow assay may be used to monitor the postoperative immune function of patients after renal transplantation.The detection of ATP content in CD4 + T cells has hinting and pre-warning function for postoperative infection,especially for severe pneumonia.

Objective To investigate the safety of young recipients undergoing living donor renal transplantation from elderly relative donors through long-term follow-up of the pathological changes. Methods According to the age of donors, 28 young recipients were divided into the observation group (n=14, elderly donors) and control group (n=14, young and middle-aged donors). The 7-year survival after renal transplantation, the serum creatinine (Scr) levels at various postoperative time points were compared between two groups. The chronic pathological injury scores of renal allograft biopsy at time-zero, postoperative 6-month and 7-year were compared between two groups. The expression levels of renal interstitial fibrosis indicators connective tissue growth factor (CTGF), transforming growth factor (TGF)-β, laminin (LN), fibronectin (FN), cell senescence indicators intercellular connexin (Cx)-43 and mammalian target of rapamycin (mTOR) at postoperative 6-month and 7-year were compared between two groups. Results The 7-year survival rates in the observation and control groups were 78.5% and 92.8% with no statistical significance (P > 0.05). In the observation and control groups, the levels of Scr were 190 and 160 μmol/L at the postoperative 7 d, and 170 and 125 μmol/L at postoperative 1 month. At each postoperative time point, the levels of Scr in the observation group were significantly higher than those in the control group (all P > 0.05). The total chronic pathological injury scores of renal transplant biopsy at time-zero in the observation group was significantly higher than that in the control group (P > 0.05), whereas the total chronic pathological injury scores at postoperative 7-year did not significantly differ between two groups (P > 0.05). Within either group, the total chronic pathological injury scores at postoperative 7-year was remarkably higher than those at time-zero and postoperative 6-month (both P < 0.05). The expression levels of CTGF, TGF-β, LN, FN, mTOR, Cx43 of renal transplant tissue at postoperative 7-year did not significantly differ between two groups (all P > 0.05). Conclusions The long-term follow-up outcomes demonstrate that the pathological changes of young recipients undergoing renal transplantation from elderly donors are similar to those from young and middle-aged donors. It is safe and feasible for young recipients to undergo renal transplantation from elderly donors in the pathological perspective.

Objective:To establish a method for rapid detection of OXA and par C resistance genes of Acinetobacter baumannii(Ab)by double nucleic acid colloidal gold strip and to develop kit.Methods:DNA of Ab was extracted by heating and boiling method.OXA and par C genes sequences of Ab were selected as target gene fragments based on NCBI.Primers were designed and labeled with 6-FAM,digoxin and biotin,respectively.Drug resistance gene detection reagents were developed,and nucleic acid gold test strips were used for rapid and visual detection.Molecular cloning and sequencing techniques were used to clone positive control samples and evaluate specificity,sensitivity and stability of kit.Results:DNA concentration and purity of Ab extracted by boiling method were good.Homology between cloned and sequenced plasmid DNA and gene sequence in GenBank database was 100%,respectively.Speci-ficity of kit was good,with only Ab showing positive results and other bacterial genera showing negative results;DNA concentration of Ab in double nucleic acid colloidal gold test strip decreased to 10-3 ng/μl,a red line still appeared,whose sensitivity was 10 times higher consistent with minimum detection limit of electrophoresis 10-2 ng/μl;test kits were tested at 3rd,6th and 9th months,and showed good stability.Conclusion:Double resistance detection kit established in this study can simultaneously detect OXA and par C resis-tance of Ab,who has advantages of high sensitivity,strong specificity,rapid and simple,and provides a new method for detection of carbapenem and quinolone antibiotic resistance of Ab.

Objective To investigate the role of cyclic GMP-AMP synthase (cGAS),a cytosolic DNA sensor,in regulating innate immune responses induced by reverse transcription intermediate of human T cell leukemia virus type 1 (HTLV-1). Methods (1)ssDNA90,the reverse transcription intermediate of HTLV-1,was transfected into HeLa cells to observe changes in the expression pattern of cGAS in transfected-HeLa cells with immunoblot assay. (2) HeLa cells were firstly transfected with cGAS-encoding plasmid and then ssDNA90 24 hours later. Real-time PCR was used to measure the expression of interferon ( IFN)-β, IFN-gamma-inducible protein 10 ( IP-10 ), regulated on activation, normal T cell expressed and secreted (RANTES) and tumor necrosis factor (TNF)-α. Immunoblot assay was performed to measure phosphorylated interferon regulatory factor 3 (IRF3) and p65. (3)cGAS expression was silenced by siRNA in HeLa and phorbol-12-myristate-13-acetate (PMA)-treated THP1 (PMA-THP1) cells and then ssDNA90 was transfect-ed into these cells 24 hours later. Real-time PCR was used to measure the expression of IFN-β,IP-10,RAN-TES and TNF-α. Immunoblot assay was performed to measure phosphorylated IRF3 and p65. Results Ex-pression of cGAS was increased in HeLa cells after ssDNA90 transfection. Compared with control cells, cGAS-transfected HeLa cells showed increased expression of IFN-β, IP-10, RANTES and TNF-α and en-hanced phosphorylation of IRF3 and p65 after ssDNA90 transfection. Compared with control cells,both HeLa and PMA-THP1 cells with silenced expression of cGAS showed impaired production of IFN-β,IP-10,RAN-TES and TNF-α after ssDNA90 transfection. Moreover,ssDNA90-induced phosphorylation of IRF3 and p65 were decreased after cGAS gene-knockdown. Conclusion cGAS might promote HTLV-1 RTI ssDNA90-in-duced innate immune responses.