Relationship of Transforming Growth Factor-β1 and Arginase-1 Levels with Long-term Survival after Kidney Transplantation / 华中科技大学学报（医学）（英德文版）

In this study,we compared the serum levels of transforming growth factor-β1 (TGF-β1),interleukin-10 (IL-10),and arginase-1 in long-term survival kidney transplant recipients (LTSKTRs) with those in short-term survival kidney transplant recipients (STSKTRs).We then evaluated the relationship between these levels and graft function.Blood samples were collected from 50 adult LTSKTRs and 20 STSKTRs (graft survival approximately 1-3 years post-transplantation).All patients had stable kidney function.The samples were collected at our institution during the patients' follow-up examinations between March 2017 and September 2017.The plasma levels of TGF-β1,IL-10,and arginase-1 were analyzed using enzyme-linked immunosorbent assays (ELISA).The levels of TGF-β1 and arginase-1 were significantly higher in the LTSKTRs than in the STSKTRs.The time elapsed since transplantation was positively correlated with the levels of TGF-β 1 and arginase-1 in the LTSKTRs.The estimated glomerular filtration rate was positively correlated with the TGF-β1 level,and the serum creatinine level was negatively correlated with the TGF-β1 level.Higher serum levels of TGF-β1 and arginase-1 were found in LTSKTRs than in STSKTRs,and we found that TGF-β1 was positively correlated with long-term graft survival and function.Additionally,TGF-β1 and arginase-1 levels were positively correlated with the time elapsed since transplantation.On the basis of these findings,TGF-β1 and arginase-1 may play important roles in determining long-term graft survival.Thus,we propose that TGF-β1 and arginase-1 may potentially be used as predictive markers for evaluating long-term graft survival.

Relationship of Transforming Growth Factor-β1 and Arginase-1 Levels with Long-term Survival after Kidney Transplantation / 华中科技大学学报（医学）（英德文版）

In this study,we compared the serum levels of transforming growth factor-β1 (TGF-β1),interleukin-10 (IL-10),and arginase-1 in long-term survival kidney transplant recipients (LTSKTRs) with those in short-term survival kidney transplant recipients (STSKTRs).We then evaluated the relationship between these levels and graft function.Blood samples were collected from 50 adult LTSKTRs and 20 STSKTRs (graft survival approximately 1-3 years post-transplantation).All patients had stable kidney function.The samples were collected at our institution during the patients' follow-up examinations between March 2017 and September 2017.The plasma levels of TGF-β1,IL-10,and arginase-1 were analyzed using enzyme-linked immunosorbent assays (ELISA).The levels of TGF-β1 and arginase-1 were significantly higher in the LTSKTRs than in the STSKTRs.The time elapsed since transplantation was positively correlated with the levels of TGF-β 1 and arginase-1 in the LTSKTRs.The estimated glomerular filtration rate was positively correlated with the TGF-β1 level,and the serum creatinine level was negatively correlated with the TGF-β1 level.Higher serum levels of TGF-β1 and arginase-1 were found in LTSKTRs than in STSKTRs,and we found that TGF-β1 was positively correlated with long-term graft survival and function.Additionally,TGF-β1 and arginase-1 levels were positively correlated with the time elapsed since transplantation.On the basis of these findings,TGF-β1 and arginase-1 may play important roles in determining long-term graft survival.Thus,we propose that TGF-β1 and arginase-1 may potentially be used as predictive markers for evaluating long-term graft survival.

Effects of intravenous cisatracurium besilate pretreatment on fentanyl-induced cough / 中国药学杂志

OBJECTIVE: To observe the effect of intravenous cisatracurium besilate pretreatment on fentanyl-induced cough (FIC). METHODS: Totally 200 American Society of Anesthesiologists class I-II patients, aged 20-60 years within normal weight, scheduled for elective surgery, were randomly assigned to receive intravenous cisatracurium besilate 0.01 mg·kg-1 (group II) or normal saline (group 1) 5 min prior to the administration of fentanyl 4 μg·kg-1 in a randomized and double-blind fashion. Any episode of cough was classified as coughing, and graded as mild (1-2), moderate (3-4), or severe (5 or more). We recorded the timing and severity of cough and monitored the vital signs and discomforts such as dizziness, tinnitus, nausea/vomiting, apnea and dvsrhythmia. RESULTS: The gender distribution showed no significant difference between two groups. The incidence of cough was 19.0% (19/100) in group II and 41% (41/100) in group I (P<0.05). The incidence of moderate and severe cough was significantly Iiigher in the group I (56.1%, 23/41) than in group 1 (31.6%, 6/19) (P<0.05). There was no significant difference between these two groups in the timing of cough. Vital signs were stable in both groups and no difference was shown in discomforts. CONCLUSION: Cisatracurium besilate pretreatment can effectively reduce the incidence and severity of FIC.

Screening and identification of high-yield poly(β-malic acid) bacterial strain / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To isolate and identify the high-yield poly-malic acid (PMLA) bacterial strains from the nature.</p><p><b>METHODS</b>Samples were collected and cultured. The high-yield PMLA bacterial strains were screened through morphological observation, qualitative PMLA tests by HPLC and ITS sequence analysis on the isolated bacterial strains.</p><p><b>RESULTS</b>A high-yield PMLA strain II 04 was isolated, the yield of PMLA of the strain reached to 26.23g/L in the rotary shaker at 25 degree for 7d. From morphological observation and ITS sequences analysis, the strain belonged to Aureobasidium pullulans, and named as Aureobasidium pullulans ZUCC-41.</p><p><b>CONCLUSION</b>A high-yield bacterial strain has been isolated from the nature and identified to be Aureobasidium pullulans.</p>

Combination of donor splenocyte transfusion with blockade of γc signal synergizes to inhibit alloreactive T-cell proliferation and induces apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The common γ chain (γc) plays a critical role in regulating proliferation, differentiation, and apoptosis of peripheral T-cells. It was previously confirmed that blocking the γc signal can successfully induce transplant tolerance in a murine model. Here we investigated the potential mechanism.</p><p><b>METHODS</b>Splenocytes from C57BL/6 mice were transfused into T-cell deficient Balb/c nude mice that were reconstituted with syngeneic wild-type T-cells labeled with 5-carboxyfluorescein diacetate succinimidyl ester (CFSE). After 24 hours, recipients received i.p. injection of mixture of anti-γc mAbs, or with isotype control IgG2a. The labeled T-cells were harvested from recipient spleens after 12 and 48 hours. T-cell proliferation and apoptosis were detected by flow cytometry.</p><p><b>RESULTS</b>T-cell proliferation was markedly inhibited and apoptotic T cells could be detected at 12 hours after the mAbs injection. Proliferation was inhibited at 48 hours, but the proportion of apoptotic T-cells was not more than at 12 hours. In the control group, however, T-cells actively proliferated and no significant apoptosis was detected at either time point.</p><p><b>CONCLUSIONS</b>The results suggested that blockade of γc signals can synergize with donor splenocyte transfusion and lead to inhibition of antigen-specific T-cell proliferation and induction of apoptotic T-cell death. This protocol may develop a novel approach to induce donor-specific tolerance.</p>

Human CD96 gene cloning, expression and identification / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.</p><p><b>METHODS</b>hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.</p><p><b>RESULTS</b>hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.</p><p><b>CONCLUSION</b>The CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.</p>

Pregnancy estrogen drives the changes of T-lymphocyte subsets and cytokines and prolongs the survival of H-Y skin graft in murine model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Estrogen as well as CD4(+)Foxp3(+) regulatory T cells were shown to have a protective role not only in maintaining maternal-fetal tolerance but also against autoimmune diseases. We aimed to investigate whether the pregnancy levels of estrogen are enough to induce transplant tolerance as to maintain fetal-maternal tolerance.</p><p><b>METHODS</b>We established H-Y skin graft transplantation in C57BL/6 ovariectomized mice that reconstituted with estrogen. Subsequently, consecutive daily estrogen injection was administrated. Tregs and the cytokines in the peripheral blood were detected by flow cytometry and ELISA pre- and post-transplant.</p><p><b>RESULTS</b>The results indicated that pregnancy levels of estrogen could promote Tregs in secondary lymphoid organs and peripheral blood (P < 0.05) but not thymus (P > 0.05). The estrogen-treated recipients accepted H-Y skin grafts for more than 35 days (median survival time (MST): (44.0 ± 1.2) days) compared with estrogen-untreated mice (MST: (23.0 ± 1.6) days) (P < 0.05). It was also observed that estrogen up-regulated the expression of Foxp3, but did not affect CD3(+)CD8(+) effector T-cells in non-transplant mice. While in the presence of H-Y antigens, the expression of Foxp3 was more significant and CD3(+)CD8(+) effector T cells were decreased significantly (P < 0.05). Meanwhile, the up-regulated IL-10 and IL-4, and down-regulated IFN-γ could be observed (P < 0.05).</p><p><b>CONCLUSIONS</b>Pregnancy levels of estrogen may promote the conversion of peripheral Tregs in secondary lymphoid organs, but show no effect on the natural Tregs production, differentiation and maturity in central lymphoid organs. Furthermore, pregnancy levels of estrogen could significantly prolong the survivals of H-Y skin grafts by the expansion of Tregs, suppression of CD3(+)CD8(+) effector T-cells and immune shift towards Th2 cytokines.</p>

Tbx3 upregulation may be one of the malignant biomarkers of breast cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of abnormal Tbx3 expression in the pathogenesis of breast cancer.</p><p><b>METHOD</b>The total RNA of 4 breast cancer cell lines and 5 normal breast samples was extracted by routine Trizol method. After reverse transcription of the total RNA into cDNA, Tbx3 mRNA expression was detected in these samples by real-time PCR. Immunohistochemistry was used to examine the differences in Tbx3 protein expression between 60 breast cancer samples and 34 normal breast tissue samples.</p><p><b>RESULTS</b>Compared to normal breast tissue samples, the breast cancer cell lines showed markedly increased Tbx3 mRNA expression. The results of immunohistochemistry demonstrated a significant upregulation of Tbx3 protein expression in the 60 breast cancer tissues in comparison with the normal breast tissues, as was consistent with Tbx3 mRNA expressions in these tissue samples.</p><p><b>CONCLUSIONS</b>The mRNA and protein expressions of Tbx3 are markedly upregulated in breast cancer cell lines and tissue samples, suggesting that Tbx3 may serve as one of the malignant biomarkers in the pathogenesis of breast cancer.</p>

Expression of TBX3 mRNA and its role in the pathogenesis and metastasis of breast cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of TBX3 gene in the pathogenesis of breast cancer.</p><p><b>METHODS</b>The total RNA of 51 fresh breast cancer tissues and the corresponding adjacent tissues were extracted and reverse transcribed into cDNA to detect the expression of TBX3 mRNA by real-time PCR. The correlation between TBX3 mRNA expression and the clinicopathologic parameters in relation to breast cancer metastasis was analyzed.</p><p><b>RESULT</b>Compared to that in the adjacent tissues, the expression of TBX3 mRNA was markedly increased in breast cancer tissues. TBX3 mRNA expression was significantly higher in metastatic breast cancer than in non-metastatic tumors.</p><p><b>CONCLUSION</b>Increased expression of TBX3 mRNA suggests the involvement of TBX3 in the pathogenesis and metastasis of breast cancer.</p>

Technology and application of simultaneous pancreas-kidney transplantation with modified enteric drainage / 中华外科杂志

<p><b>OBJECTIVE</b>To report the modified technique and the short-term results of simultaneous pancreas-kidney transplantation (SPK) with the enteric drainage (ED) of exocrine secretions.</p><p><b>METHODS</b>From June 2000 to August 2006, thirty-eight patients with diabetes complicated with uremia underwent SPK. The pancreas graft was placed intraperitoneally with exocrine secretions drained into the proximal jejunum without Roux-en-Y procedure. The mean cold ischemic times of pancreas and kidney were (10 +/- 2.0) h and (7 +/- 2.0) h, respectively. Quadruple immunosuppressive therapy with antilymphocyte globulin or anti-CD25 monoclonal antibody, tacrolimus, mycophenolate mofetil and steroids was adopted except one patient.</p><p><b>RESULTS</b>The 6-month survival rates of patients and grafts were both 97.4% after transplantation. All patients achieved insulin-free euglycemia at (7 +/- 6.9) d postoperative except one. For preoperative patients, mean fasting insulin and C-peptide values were (9 +/- 8.1) mU/L and (6 +/- 4.5) mU/L. After operation, fasting insulin and C-peptide values of patients were (12 +/- 5.8) mU/L and (6 +/- 4.7) mU/L, respectively, which peaked to an insulin level of (57 +/- 43.0) mU/L and a C-peptide level of (11 +/- 6.8) mU/L with stimulation. There were eight cases of delayed renal graft function. All other patients achieved immediate renal graft function. No graft losses occurred due to leakage or intra-abdominal infection. The most common surgical complications were wound infection (n = 12), enteric anastomostic hemorrhage (n = 5) and perirenal hemorrhage (n = 2). Three patients (7.9%) had been reoperated for the reasons of intra-abdominal hemorrhage and perirenal hemorrhage.</p><p><b>CONCLUSIONS</b>SPK is an effective treatment option for selected patients with diabetes mellitus and approaching end-stage renal disease. Enteric exocrine drainage by direct side-to-side anastomosis (without Roux-en-Y) seems to be a simple and reliable technique.</p>

The phagocytosis of apoptotic allogeneic lymphocytes by recipient liver antigen-presenting cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To track the location of the transfused apoptotic allogeneic lymphocytes and asses the process of their accumulation and phagocytosis removal as consequences on allograft tolerance in recipient mice.</p><p><b>METHODS</b>Donor spleen lymphocytes were labeled by CFSE and induced to apoptosis by dexamethasone incubation. After purification by anti-annexin V-conjugated magnetic beads isolation, apoptotic lymphocytes were transfused into recipient mice through the tail veins. Tissue samples from various organs were taken at various time points to analyze the fates of the apoptotic allogeneic lymphocytes and the phagocytosis of them by organ resident APCs.</p><p><b>RESULTS</b>Using fluorescent microscopy and flow cytometry, after the apoptotic cells were recognized and uptaken, the largest amount of labeled cells were accumulated in the livers and disappeared within not more than 12 hours. Recipient liver APCs were highly efficacious in phagocytosis of apoptotic allogeneic lymphocytes; the removal was completed within 15 minutes after incubation. LSEC, KC and LDC all phagocytosized the apoptotic allogeneic lymphocytes but with significantly different rates. Considering the numbers of those cells in a normal liver, it could be calculated that LSEC and KC had greater effects in this activity.</p><p><b>CONCLUSIONS</b>The liver deserves foremost attention for study of the mechanism of allografts tolerance induced by pre-transfusion of apoptotic donor spleen lymphocytes. LSEC and KC are the main functional APCs to the alloantigens.</p>

Pretreatment of dondor dendritic cells with Ad-IL-12p35siRNA on the survival of allograft recipients / 中华器官移植杂志

Objective To explore the effects of donor dendritic cells(DC)treated with Ad-IL- 12p35siRNA on the survival of allogragft recipients.Methods The recombinant adenoviral vectors carrying IL-12p35siRNA and HKsiRNA were transfected into bone marrow derived DC of BALB/C murine.C57BL/6 recipients were infused with DG(Ad-IL-12p35siRNA DC,Ad-HKsiRNA DC and control DC)from BALB/C donors 7 days before cardiac allograft,the survival time of murine and the change of T_H 1 and T_H2(IL-2,IL-4,IL-10 and IFN-?)cytokine were observed.Results The survival time of p35 group(20.17?2.71)days was longer than that of control group(7.81?1.61)days and HK group(7.17?1.60)days.The concentration of IL-2 and IFN-?in p35 group were significantly lower than those of control group and HK group,otherwise were the concentration of IL-4 and IL-10. Conclusion Pretreatment of dondor dendritic ceils with Ad-IL-12p35siRNA could prolonged cardiac allograft survival in recipicents.

Study the mechanisms and inducing transplantation immune tolerance of FTY720 / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the operational mechanisms and potential approach to inducing transplantation immune tolerance of FTY720.</p><p><b>METHODS</b>Mouse splenocytes were incubated with FTY720, then the DNA was extracted and analyzed using gel electrophoresis. Hearts of inbred BALB/c (H-2(d)) mice were transplanted heterotopically in C57BL/6 (H-2b) mice. Recipients were randomly divided into six groups. Group-1 (n = 6) was the nil-treated control. Groups-2, 3 and 4 were given FTY720 at the dose of 3 mg.kg(-1) by oral gavage once a day with different time courses. Group-2 (n = 14) were administrated from 3 days before transplantation (day-3) to the 11th day after the transplantation (day 11); Group-3 (n = 6) from day 0 to day 14; Group-4 (n = 6) from day-3 to day 0. Group-5 (n = 5) and 6 (n = 5) were treated with Cyclosporine A (10 mg.kg(-1)) and 40-O-(2-hydroxyethyl)-rapamycin (RAD) (3 mg.kg(-1)) respectively by daily gavage from day 3 to day 11. The long survivors (> 100 d) in Group-2 were detected with their IL-4 and IFN-gamma levels and their tolerant state was challenged with second graft: the donor type skin.</p><p><b>RESULTS</b>Apoptosis changes of the mouse splenocytes incubated with FTY720 was showed by typical DNA ladders. The median survival time (MST) of Group-1 was 8 d. MST of Group-2 was 55 d and grafts in six mice survived more than 100 d. MST of Group-3 was 16.5 d. Group-4 has a MST of 14 d with one case exceeded 100 d. MST of Group-5 and 6 were 10 d and 13 d respectively. Long survivors of Group-2 can accept donor-type skin graft and the level of IL-4 in their serum is up-regulated while IFN-gamma remained stable.</p><p><b>CONCLUSIONS</b>Pretreatment of FTY720 bring about effect on the early events of transplantation immune responses. This effect might be mediated by apoptosis induction in lymphocytes using this drug. We originally designed the regime of FTY720 monotherapy, which started pre-operationally and maintained for a short period of time, and induced stable tolerance the allo-graft in mouse.</p>