Expression of costimulatory molecule inducible costimulator and coinhibitory molecule programmed death-1 in patients with myasthenia gravis / 中华神经科杂志

Objective To explore the immunopathological mechanism for the imbalance between the positive signal mediated by inducible costimulator (ICOS) and the negative signal mediated by programmed death-1 (PD-1) in patients with myasthenia gravis (MG).Methods Eighty-two patients with MG,56 healthy controls (HC) and 20 non-MG (NMG) patients,collected in the First Affiliated Hospital of Suzhou University from February 2014 to December 2016,were chosen to participate in the study.The expression of ICOS and PD-1 on peripheral blood mononuclear cells was detected by immuno-fluorescence staining and flow cytometry.The levels of soluble programmed death-1 (sPD-1),soluble programmed death ligand 1 (sPD-L1),IL-4 and other cytokines were detected by enzyme-linked immunosorbent assay.Results (1) Flow cytometry analysis:The co-expression of PD-1,ICOS on CD4 + T cells from MG group (9.64％ (8.82％)) was higher than in HC (1.81％ (2.10％),Z =-7.389,P ＜0.05) and NMG group (2.86％ (1.49％),Z =-4.636,P ＜ 0.05).The expression of ICOS on CD4 + T cells,ICOS ligand (ICOSL) on CD14+ monocytes and CD19+ B cells were increased in MG group comparing with that of the control groups.The proportion of PD-1 + CD4 + T cells (MG group 16.82％ (10.66％),HC 9.34％ (9.18％),Z =-4.345,P＜0.05;NMG group 7.07％ (3.40％),Z=-4.594,P＜0.05) and PD-1 Ligand (PD-L1) + CD14+ monocytes was higher in MG patients.All of these were detected by flow cytometry.(2) ELISA analysis:Serum sPD-1 expression significantly increased in MG group compared with that in the control groups (MG group (1.87 ± 0.64) ng/ml,NMG group (1.49 ± 0.70) ng/ml,t =2.04,P ＜ 0.05;HC (1.05 ± 0.50)ng/ml,t =2.08,P ＜ 0.05),while for serum sPD-L1,there was no significant difference between MG and control groups.(3) Serum cytokines detection:The expression of IL-4 was increased in MG patients (MG group (61.88 ±5.15) pg/ml,HC (32.03 ±1.84) pg/ml,t=2.50,P＜0.05;NMG group (42.62± 3.31) pg/ml,t =2.34,P ＜0.05),and there was a negative correlation between the expression of sPD-1 and the concentration of IL-4.Conclusions The increased expression of PD-1 + ICOS + CD4 + T cells suggested the subset involved in the pathological progress of MG.sPD-1 might disturb the ligation of PD-1 on T cells and PD-L1 on antigen presenting cells,while the ligation of ICOS and ICOSL passed positive signal,leading to over activity of the subsets and the progression of disease.

Expressions and clinical significance of T cell immunoglobulin and mucin-domain-containing molecule-3 and its ligand in the peripheral blood of patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression of T cell immunoglobulin and mucindomain-containing molecule-3 (TIM3) on PBMCs,and the plasma concentrations of soluble forms of Galetcin9 and their clinical relationship with rheumatoid arthritis (RA).Methods Peripheral blood samples were collected from 39 patients,25 osteoarthritis (OA) patients and 20 healthy subjects (HC).The expressions of TIM3 on peripheral blood mononuelear cells (PBMCs) were detected by flow cytometry.The concentrations of soluble Galetcin9 were assessed by enzyme linked immunosorbent assay (ELISA).And the relationship between their expression levels and clinical manifestations were analyzed.Levene F test was used for statistical analysis,normal distribution data were compared by t test and Pearson correlation analysis,while Mann-Whitney U test and Spearman correlation analysis were used for non-normal distribution data.Results The expression of TIM3 on CD4+ T cells was significantly higher than that of the HC [(14.7±3.2)％ vs (5.1±0.8)％,t=2.339,P=0.022 7],while there was no statistical difference between the RA group and the OA group [(14.7±3.2)％ vs (5.8±0.4)％,t=1.928,P=0.058 9].The expression of TIM3 was significantly correlated with the concentration of RF in the serum and the corresponding DAS28 score (r=0.325 8,P=0.043 0;r=0.407 5,P=0.010 0).The expression of TIM3 on CD8 + T cells in RA patients was significantly higher than that in the HC and OA [(21.1±3.4)％ vs (8.3±1.5)％,t=2.531,P=0.0142;(21.1±3.4)％ vs (10.7±1.0)％,t=2.314,P=0.024 0] which was significantly correlated with the concentration of RF in serum and the corresponding DAS28 score (r=0.451 5,P=0.003 9;r=0.524 1,P=0.000 6) as well.However,the expression of TIM3 on CD56+ NK cells was not significantly different from that of either OA or HC[(56.4±3.4)％ vs (50.6±3.8)％,t=1.047,P=0.299 8;(56.4± 3.4)％ vs (56.1±3.4)％,t=0.048,P=0.961 9],the concentration of serum RF and the corresponding DAS28 score were not significantly related.We also found that plasma Galectin 9 concentrations in RA patients were significantly higher than those of OA patients [(4.24±0.22) ng/ml vs (3.15±0.18) ng/ml,t=3.187,P=0.024] and healthy subjects [(4.24±0.22) ng/ml vs (2.55±0.14) ng/ml,t=5.567,P＜0.01],which was correlated with RF and DAS28 (r=0.479 2,P=0.002 0;r=0.353 0,P=0.027 5) while there was no correlation with CRP (r=0.176 3,P=0.283 1).Conclusion The upregulated expressions of TIM3 on peripheral lymphocytes and the high levels of plasma concentration of soluble Galectin 9 are closely correlated with the severity of the disease,suggesting that TIM3/Galectin 9 pathway may play a critical role in the pathogenesis of RA.

Cortistatin expression in renal tissue of humans and IgA nephropathy patients / 军事医学

Objective To explore the condition of cortistatin (CST)expression in human renal tissue and the changes in the level of CST in IgA nephropathy (IgAN)of different degrees.Methods Ten tumor adjacent normal renal tissue samples were collected.The mRNA and protein expressions of CST in human renal tissue were detected by reverse transcription-polymerase chain reaction (RT-PCR)and Western blotting,respectively.Immunohistochemisty (IHC)was performed to locate the expression of CST in renal tissue.According to the grading system of Lee et al,IgAN was divided into three groups:grade Ⅰ -Ⅱ (group A),grade Ⅲ -Ⅳ (group B),and grade Ⅴ (group C),and ten renal biopsy tissue samples were collected for each group.IHC was performed to detect the change in the level of CST in normal and IgAN renal tissue of different degrees.The effect of clinical indices on the level of CST in IgAN renal tissue was assessed by multiple linear regression analysis.Results RT-PCR and Western blotting showed that CST was expressed in renal tissue and IHC showed that CST was expressed on renal tubular epithelial cells.In IgAN,the higher the pathological grade was, the higher the expression of CST in renal tubules was.Multiple linear regression analysis showed that the pathological grade was associated with the expression of CST in renal tissue (r =0.875,P <0.01).Conclusion CST may participate in the inflammatory reaction of IgAN pathological injury and exert anti-inflammation effects.

Research progress in urine biomarkers in IgA nephropathy / 军事医学

IgA nephropathy ( IgAN) , which is characterized by the presence of IgA deposits in the glomerular mesangi-um, is the most common form of primary glomerulonephritis in the world .Some studies have revealed that the expression and changes of certain factors involved in IgAN pathology lesion in urine can provide facilitate the diagnosis , assessment and prognosis of IgAN .Studies on urine biomarkers in recent years were reviewed in this paper .

Correlation of expression of Oct-4 and tumorigenesis and prognosis of colorectal cancer / 国际肿瘤学杂志

Objective To study dynamic expression of Oct-4 during development of colorectal cancer (CRC) and investigate the association between Oct-4 expression and prognosis of patients with CRC.Methods CRC tissues, matched non-tumor tissues and colonic polyp tissues were collected and Oct-4 expressions were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), flow cytometry analysis (FCM) and immunohistochemistry (IHC).Medical records of patients with CRC were reviewed.Clinicopathological analysis was performed to assess the association between Oct-4 expression and certain chnicopathological parameters.Kaplane-Meier survival curve was conducted to evaluate association between Oct-4 expression and survival time of patients with CRC.Results The results of qRT-PCR showed that the relative expressions of Oct-4 mRNA in matched non-tumor, colonic polyp and CRC tissues were (0.23 ±0.16) × 10-5, (1.19 ± 0.76) × 10-5 and (3.79 ± 1.94) × 10-5 respectively, with a significant difference (F =8.633, P =0.001).IHC analysis displayed that the positive expression ratios of Oct-4 in matched non-tumor tissues, colonic polyp tissues and CRC tissues were 4.43％, 12.68％ and 40.51％ respectively, and a significant difference was discovered among them (x2 =66.311, P ＜ 0.001).Oct-4 expre-ssion in CRC was significantly correlated with pathological stage (x2 =7.248, P =0.007), lymph node metastasis (x2 =4.888, P =0.027), distant metastasis (x2 =5.732, P=0.017) and TNM stage (x2 =4.174, P=0.041).Kaplan-Meier survival curve analysis demonstrated that Oct-4 positive cases had a shorter median survival time (37.0 months) compared with Oct-4 negative cases (76.0 months), with a significant difference (x2 =14.050, P =0.001).Conclusion The expression of Oct-4 is step-wise increased during development of CRC.The abnormal expression of Oct-4 may play certain roles in the development of CRC.Oct-4 may be an important biomarker for the clinical diagnosis, assessment of disease and prognosis in CRC.

Microsurgery for cerebellopontine angle epidermoid cyst presenting with trigeminal neuralgia / 中国医师进修杂志

Objective To summarize clinical diagnosis and microsurgical treatment method of cerebellopontine angle epidermoid cyst presenting with trigeminal neuralgia.Methods Thirty-seven cases of cerebellopontine angle epidermoid cyst presenting with trigeminal neuralgia treated by microneurosurgery procedure were analyzed retrospectively.Results Complete resection was achieved in 31 cases,and subtotal was achieved in 6 cases,with no operative mortality.Postoperatively,the symptom of trigeminal neuralgia disappeared in 28 cases,was eased obviously in 6 cases,and was eased in 3 cases.The symptom of dizziness in all 6 cases vanished,and hearing in 3 cases was improved.Three cases appeared cerebrospinal fluid leakage,and 4 cases appeared oral herpes.Symptom in 1 case aggravated after 6 months,and oral medicine was ineffective.The pain symptom disappeared after reoperation.Conclusions The suboccipital retrosigmoid approach microneurosurgery is the principal treatment method for cerebellopontine angle epidermoid cyst presenting with trigeminal neuralgia.Microsurgical technique combined with surgical skill is the guarantee for increasing total removal rate of tumors and decreasing complications.

Clinical significance of ICOS and ICOSL expression in patients with rheumatoid arthritis / 中国免疫学杂志

Objective:To investigate the expression of inducible costimulatory ( ICOS) and inducible costimulatory ligand ( ICOSL) on peripheral blood mononuclear cells ( PBMCs ) and their clinical relationship with rheumatoid arthritis ( RA ) patients.Methods:Peripheral blood samples were collected from 85 RA patients and 50 HC in this study.Expression of ICOS and ICOSL on PBMC from the subjects were detected by flow cytometry and real-time polymerase chain reaction( RT-PCR).The alteration of ICOS and ICOSL were observed after hormone therapy in 15 patients with RA and the relationship between their expression level and patients′clinical manifestations were analysed.Results:The ICOS and ICOSL mRNA level of RA patients′PBMCs were significantly higher than that in HC.The expression level of ICOS on CD4+T cells was higher than than that in HC[(7.08±4.72)% vs (3.01+1.39)%,P<0.0001].The expression of ICOSL on monocytes[(5.77±3.45)%vs (3.64±1.43)%,P<0.05] and B cells [(5.78± 4.52)%vs (3.97±1.63)%,P<0.05] were significantly elevated in RA patients.In RA patients with active disease,however,ICOSL expression on monocytes and B cells were increased as compared with those in inactive RA patients [ ( 5.45 ±3.50 )% vs ( 4.04 ± 1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Furthermore,after receiving immunosuppressive therapy, the expressions of ICOS and ICOSL were notably reduced as compared with pre-therapy levels on PBMCs from patients [ ( 5.45 ±3.50)%vs (4.04±1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Conclusion:The high levels of ICOS and ICOSL expression were closely correlated with the degree of disease and therapeutic response,suggesting that ICOS/ICOSL pathway may play a critical role in pathogenesis of RA.

Inhibitory effect of tight junction protein on experimental corneal neovascularization / 中华实验眼科杂志

Background Corneal neovascularization (CNV) is one of the causes of corneal blindness.Studies showed that zonula occludens-1 (ZO-1) can inhibit pathological angiogenesis through physical barrier formed by tight junction structure.However,whether ZO-1 plays a role in CNV is unclear.Objective The aim of this study was to explore the effect of ZO-1,a tight junction protein on experimental CNV.Methods The CNV models were established in the left eyes of 24 clear male BALB/c mice aged 7-8 weeks by putting NaOH filter paper in the center of corneas for 15 seconds (15 s group) or 40 seconds (40 s group).CNV was examined and evaluated under the slit lamp microscope,and the expression of ZO-1 mRNA in the corneas were detected and compared by reverse transcription PCR (RT-PCR) between the two groups 2 weeks after modeling.In addition,54 models created by the same method were assigned to 3 groups according to randomized number table,0.2％ hyaluronic acid (HA),antiZO-1 neutralizing antibody (10 mg/L) +0.2％ HA and mouse hypoxia inducible factor-1α (HIF-1α) recombinant protein (5 mg/L)+0.2％ HA were topically administrated in the mice three times a day for 1 week after modeling respectively.The corneas were extracted 2 weeks after application of the drugs.Expression of CD31 in the CNV was assayed to calculate the number and the area of CNV by immunohistochemistry.The expression of VEGF mRNA in the corneas was detected by RT-PCR.The percentages of macrophage-specific F4/80 positive cells and neutrophilsspecific Ly-6G positive cells were calculated to evaluate the infiltrations of inflammatory cells in the corneas by flow cytometry.Results In 2 weeks after alkali burn of corneas,the number of severe CNV was more in the 40 s group than that in the 15 s group (x2 =6.032,P=0.049),and the expression level of ZO-1 mRNA was lower in the 40 s group than that in the 15 s group (1.15±0.08 versus 1.53±0.04) (t=4.157,P=0.014).CD31 positive cell number was more and the staining area was larger in the ZO-1 antibody group and HIF-1α positive control group than those in the 0.2％ HA group (cells:t=-129.590,-226.820,both at P=0.000;area:t =-5.310,-8.840,both at P=0.000).The relative expressions level of vascular endothelial growth factor (VEGF) mRNA was 1.33±0.10 and 1.46±0.11 in the ZO-1 antibody group and HIF-1 α positive control group respectively,which were significantly higher than 0.93±0.06 of the 0.2％ HA group (t =-5.820,-7.284,both at P =0.000).The percentages of positive cells in the ZO-1 antibody group and HIF-1α positive control group were significantly increased in comparison with the 0.2％ HA group for F4/80 (t =-16.750,-17.480,both at P =0.000) and for Ly-6G (t =-21.450,-27.680,both at P=0.000).Conclusions Alkali burn induced CNV downregulates the expression of ZO-1 mRNA.Administration of ZO-1 antibody causes the rise of VEGF mRNA in CNV and the infiltration inflammation cells,which suggests that the influence of ZO-1 on CNV is associated with the expression of VEGF.

Experimental study on in vitro preparation of adoptive immunotherapy cells induced by CD40 in combination with cytokines / 中国免疫学杂志

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.

Analysis of pathological data of renal biopsy at one single center in China from 1987 to 2012 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In China, the prevalence of chronic kidney disease has increased significantly. Many studies shows that the spectrum of kidney disease had changed in recent years. We retrospectively analyzed the pathological types of renal biopsy and its spectrum change at the General Hospital of the Chinese People's Liberation Army from December 1987 to December 2012, in order to offer new supporting evidences for further specifying the distribution of renal pathological types in China.</p><p><b>METHODS</b>According to the "Revised Protocol for the Histological Typing of Glomerulopathy" (WHO, 1995), pathological diagnosis of renal biopsy was classified, detection rate of each pathological type was summarized (i.e., percentage of total renal biopsy cases), study period was divided at an interval of 5 years, and age-stratified distribution change of main pathological types was analyzed.</p><p><b>RESULTS</b>The proportion of pathological types in 11 618 cases of renal biopsy was as follows: primary glomerulonephritis (PGN, 70.7%), secondary glomerulonephritis (SGN, 20.7%), tubular-interstitial nephropathy (4.0%), hereditary/rare nephropathy (0.3%), end-stage renal disease (0.9%), and unclassified renal disease (3.3%). Among PGN, there was IgA nephropathy (IgAN, 37.0%), membranous nephropathy (MN, 11.8%), mesangial proliferative glomerulonephritis (MsPGN, 8.9%), minimal change disease (MCD, 6.6%), and focal segmental glomerulosclerosis (3.9%). Among SGN there was lupus nephritis (LN, 5.5%), Henoch-Schönlein purpura glomerulonephritis (5.3%), hepatitis B virus-associated nephritis (HBVAN, 3.03%), diabetic nephropathy (2.2%), and hypertension/malignant hypertension-associated renal damage (1.9%). Pathological data were analyzed from 1987-1992 to 2008-2012 (after age adjustment). Detection rate of IgAN tended to rise (P < 0.001). Detection rates of MN and MCD rose significantly (P < 0.001), but detection rate of MsPGN dropped significantly (P < 0.001). Among SGN, detection rate of HBVAN tended to drop (P < 0.001).</p><p><b>CONCLUSION</b>In China, PGN was the most common glomerulopathy (mostly IgAN), LN was the most common SGN, and detection rate of MN and MCD rose significantly.</p>

Expression characteristics and significance of miR-338-5p in renal transplant pa-tients / 中国免疫学杂志

To explore the serum miR-338-5p expression characteristics in renal transplant recipients ,and the role of regulating BAFF signal ,then investigate its biological significance.Methods:Healthy volunteers were enrolled as control group.Serum miR-338-5p was detected by real-time PCR;soluble BAFF was detected by ELISA;anti-HLA-Ⅰantibody,anti-HLA-Ⅱ antibody and anti-MICA antibody were detected by liquid chip technology.SPSS17.0 software was applied.t-test was used to compare the means of two independent samples;Paired samples t-test was used to compare the means of two paired samples;Spearman method and Pearson method were used to analyse the correlation;P<0.05 was considered to be statistically significant.Results: Compared with healthy controls,serum miR-338-5p in renal transplant recipients decreased significantly (P<0.001),while serum BAFF increased significantly (P<0.01).Serum miR-338-5p levels within 1 year post-transplantation were significantly lower than that of more than 1 year post-transplantation (P<0.01);Serum miR-338-5p levels within 3 years post-transplantation were significantly lower than that of more than 3 years post-transplantation (P<0.01);To all research objects,serum miR-338-5p was significantly negatively correlated with serum BAFF (r=-0.51,P<0.001),and serum miR-338-5p was significantly negatively correlated with anti-HLA-Ⅱ antibody(r=-0.322, P<0.05);Serum miR-338-5p within 3 years was significantly negatively correlated with anti-HLA-Ⅱantibody (r=-0.423,P<0.05), and serum miR-338-5p within 3 years was significantly negatively correlated with anti-MICA antibody(r=-0.411,P<0.05);Serum miR-338-5p more than 3 years was significantly positively correlated with anti-MICA antibody(r=0.486,P<0.05),and Serum miR-338-5p more than 3 years was significantly positively correlated with anti-HLA & MICA antibody(r=0.578,P<0.01).Conclusion:miR-338-5p may directly or indirectly target BAFF signal ,and participate antibody mediated immune response by regulating its target genes and interfere with the long-term survival of transplanted renal.

Expressions of programmed death-1 and programmed death ligand on the surface of peripheral blood lymphocytes in patients with tuberculosis / 中华传染病杂志

Objective To describe the expressions of programmed death-1 (PD-1) and its ligand PD-L1 on the surface of peripheral blood lymphocytes in patients with tuberculosis.Methods A total of 77 cases of pulmonary tuberculosis were recruited,of which 27 were single infection,41 were coincident with bacterial or fungal infection and 9 patients with diabetes mellitus.Twenty-nine healthy donors were also enrolled as control group.The expressions of PD-1/PD-L1 on the peripheral blood lymphocytes and mononuclear cells were detected using immunostaining and flow cytometry.Collected data were analyzed with t-test statistics.Results Among the three groups of tuberculosis including pulmonary tuberculosis,pulmonary tuberculosis coincident with infection and pulmonary tuberculosis with diabetes mellitus,the percentages of CD4+ CD25+ T cells as well as CD4+ CD25high T cell subsets were both significantly higher than those in healthy controls (t=4.892,4.635,4.974,5.407,4.660,5.279,all P＜0.01).The expression of PD-1 was up regulated on the surface of CD8+ T cells in the groups of tuberculosis as compared with the control group (t=6.392,8.249,7.072,all P＜0.01).The proportions of PD-L1 expressed on the monocytes in each group were (42.51 ± 7.54) ％,(49.42± 6.29) ％ and (48.46 ± 14.58)％,respectively.The difference was significant in contrast to the control group,which was (17.91 ±3.03)％ (t=5.168,6.854,5.665,all P＜0.01).PD-L1 expression was also up-regulated on B cells in each group and much higher than that of the control group (51.51±7.32)％,(50.85±7.09)％,(55.66±14.29)％ vs (40.11±4.25)％ (t=2.562,2.046,2.766,all P＜0.05).Conclusion PD-1/ PD-L1 co-inhibitory pathway could be closely related to the development of tuberculosis and its complications,which is involved in the attenuation of anti-tuberculosis and anti infection immune responses.

High concentration uric acid regulates endothelial function via miR-155 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate high-concentration uric acid-induced endothelial dysfunction mediated by miR-155.</p><p><b>METHODS</b>Human umbilical vein endothelial cells (HUVECs) were incubated with 600 µmol/L uric acid for 24 and 48 h, and eNOS expression and NO content in the cell culture were measured. The target genes of miR-155 were predicted using on-line analysis software and validated by dual-luciferase system. Real-time PCR was used to detect the expression of miR-155 in endothelial cells incubated with high-concentration uric acid. The effect of miR-155 on endothelial dysfunction was assessed by transfection of its inhibitor into HUVECs.</p><p><b>RESULTS</b>The expression of eNOS and NO secretion decreased obviously in HUVECs incubated with 600 µmol/L uric acid. MicroRNA on-line analysis software and dual luciferase reporter experiments suggested that the level of eNOS translation was directly regulated by miR-155. The expression of miR-155 in endothelial cells was upregulated after stimulation with high-concentration uric acid, and was inhibited by transfection of miR-155 inhibitor. Expression of eNOS and secretion of NO were elevated in endothelial cells transfected with miR-155 inhibitor after incubation with high-concentration uric acid.</p><p><b>CONCLUSION</b>High-concentration uric acid can down-regulate eNOS expression via miR-155 to induce endothelial dysfunction.</p>

Involvement of miR-338-5p in antibody-mediated renal allograft rejection by targeting TRAF3 / 中华器官移植杂志

Objective To explore the significantly differentially.expressed microRNAs during antibody-mediated renal allograft rejection.Method MicroRNA array assay was used,and the obtained data were analyzed by bioinformatics analysis.The obtained significant microRNAs were further analyzed to forecast the targeted genes in the common database,then experimental means were used to testify the targeted genes.Result During the antibody-mediated renal allograft rejection,the significantly over-expressed microRNAs were miR-200c,miR-200b,miR-30c,miR-30b and miR-30e+,etc.The significantly down-expressed microRNA was miR-338-5p.The bioinformatics analysis results indicated that TRAF3 was the targeted gene of miR-338-5p,which was testified by real time PCR,immunohistochemical assay and fluorescence reporter assay.Conclusion miR-338-5p anticipated in the antibody-mediated renal allograft rejection by targeting TRAF3.

MicroRNA and renal carcinoma / 国际肿瘤学杂志

MicroRNAs (miRNAs) are evolutionary conserved non-coding RNA molecules.They involve in a variety of biological processes such as regulating the individual development,cell division,differentiation,apoptotic and fat metabolism through post-transcription pathway.It has been shown that miRNAs play important roles in the development and progression of many tumors,including of renal carcinoma,miRNAs play functions similar to oncogenes or anti-oncogenes in the development and progression of renal carcinoma by regulating the signaling pathway of its target genes involving in.

Prognosis of patients with diffuse axonal injuries following continuous lumbar cistern drainage / 中华创伤杂志

Objective To observe influence of continuous lumbar cistern drainage on levels of amyloid β-peptide (Aβ) subtype (Aβ1-42) in cerebrospinal fluid and plasma in patients with diffuse axonal injury (DAI) and investigate its clinical significance.Methods Eighty-one DAI patients were enrolled and randomized into treatment group (42 cases) and control group (39 cases).Patients in control group received simple conventional therapy,while the patients in treatment group received not only conventional therapy but 14 days of continuous lumbar cistern drainage.Levels of Aβ1-42 in cerebrospinal fluid and plasma were detected by ELISA assay before therapy and at 1,5,9,and 14 days after therapy.Prognosis was assessed using GOS at 6 months after therapy.Results Levels of Aβ1-42 in cerebrospinal fluid and plasma showed a decrease in the first place and a gradual decrease afterwards in both groups,but a bigger and earlier drop of Aβ1-42 levels was observed in treatment group.Two groups showed significant difference of Aβ1-42 levels at day 14 (P ＜ 0.05).At 6 months after therapy,GOS score between treatment and control groups was (4.1 ± O.5) and (3.4 ± 0.3) points respectively (P ＜0.05).Conclusion Continuous lumbar cistern drainage improves the prognosis of DAI and this may relates to the decrease of Aβ1-42 levels in cerebrospinal fluid and plasma.

Therapeutic potential of non-adherent bone marrow-derived stem cells for acute radiation injury / 中国组织工程研究

BACKGROUND: Effective treatment for severed acute radiation sickness (over 8 Gy) has not been obtained at present. Mesenchymal stem cells, which are shown to secrete hematopoietic cytokines and support hematopoietic progenitors, play an important role in cute radiation sickness. OBJECTIVE: To investigate the therapeutic potential of non-adherent bone marrow-derived stem cells in the treatment of acute radiation injury induced by 8.5 Gy X-ray irradiation, as wel as the mechanisms involved. METHODS: Non-adherent marrow-derived stem cells from the long bone of fetal limbs were col ected for analyzing surface antigens, cel cycle, osteogenic and adipogenic differentiation potential, and expressions of vascular endothelial growth factors and Annexin A2. After being exposed to 8.5 Gy total body irradiation, BALB/C mice were randomly assigned into transplantation group and control group. Mice in the transplantation group were given 3×106 CFDA-SE labeled human non-adherent bone marrow-derived stem cells, and those in the control group were given 0.3 mL normal saline. Then, the survival rate, peripheral white blood cells at different time, pathologic change and angiogenesis of the bone marrow were observed. RESULTS AND CONCLUSION: After X-ray irradiation, transplanted non-adherent mesenchymal stem cells appeared to have a homing to the site of injury. The survival rate of mice in the transplantation group was much higher than that in the control group. Compared with the control group, the white blood cells in the transplantation group decreased more slowly while recovered more rapid: the nadir appeared at day 14 after transplantation while it recovered within 30 days. The bone marrow of mice in the transplantation group regenerated more actively and had more hematopoietic islands than those in the control group on day 21. In addition, bone marrow angiogenesis of the transplantation group was more obvious than that of the control group. In conclusion, human fetal non-adherent bone marrow-derived stem cells could promote bone marrow angiogenesis in a mouse model of acute radiation injury, through which they could play an important role in tissue regeneration of acute radiation injury.

Metformin inhibits proliferation of human pancreatic cancer cell line Patu8988 / 中华内分泌代谢杂志

The proliferation of Patu8988 cells pre-treated with metformin for 72h was measured by CCK-8 assay.Cell cycle was analyzed by flow cytometry.The expression of related genes was detected by RT-PCR.Metformin suppressed the proliferation of Patu8988 cells in a dose-dependent manner( r=0.994,P＜0.05 ).The proportion of cells at G0/G1 stage was increased while that of G2/M stage decreased (P＜0.05 ).The expressions of MMP-3,cyclinD1,p53 were down regulated and that of Bax was up regulated.The results show that metformin may inhibit the proliferation of Patu8988 cells,via blocking the cell cycle and affecting the expression of related genes.

The therapeutic effects of two different ways of autologous bone marrow mesenchymal stem cells transplantation for treating early spinal cord injury / 中国医师进修杂志

ObjectiveTo compare the short-term efficacy and safety of autologous bone marrow mesenchymal stem cells transplantation on patients with early spinal cord injury by subarachnoid injection and by intravenous injection.MethodsNinety-six patients with early spinal cord injury were hospitalized and treated from November 2006 to March 2010.Thirty-eight cases (subarachnoid transplantation group) got transplantation by subarachnoid injection,32 cases (intravenous transplantation group) got transplantation by intravenous injection,26 cases (control group) were hospitalized in the same period but not transplanted.The motor and sensory functions of all three groups were evaluated according to the score standard developed by American Spinal Injury Association(ASIA) before treatment and at the first,the third,the sixth month after treatment.Meanwhile,routine blood test,coagulation,biochemical items and tumor markers were also examined in follow-up.ResultsThe motor and sensory function of three groups had different degree of recovery at the first month after treatment,and sensory function recovered muchsignificantly,but the comparison among three groups had no statistical significance.The scores of motor function increased in three groups at the third month after treatment,but still had no statistical significance (P＞ 0.05).The scores of sensory function of subarachnoid transplantation group[(130.9 ±41.6) scores] and intravenous transplantation group [ (131.2±22.7 ) scores ] increased obviously,and had significant difference compared with control group [ (109.3±36.4) scores] (P ＜ 0.05),but there were nosignificant difference between subarachnoid transplantation group and intravenous transplantation group (P ＞ 0.05).The scores of sensory and motor function of control group didn't increase obviously at the sixth month after treatment,while the scores of subarachnoid transplantation group and intravenous transplantation group gradually improved and had statistical significance compared with control group(P ＜ 0.05).The scores of sensory function was higher in subarachnoid transplantation group[ (151.6±46.9) scores ] than that in intravenous ransplantation group [(134.6 ±40.7) scores] (P ＜0.05).There were no obvious abnormality in the results of followed-up examination.Conclusions The safety and short-term efficacy of autologous bone marrow mesenchymal stem cells transplantation in treating early spinal cord injury by subarachnoid injection and intravenous injection is certified.The subarachnoid injection is better than intravenous injection,but the long-term efficacy need furter study.

Expression and clinical significance of phosphorylation of mammalian target protein of rapamycin in human glioma / 中国医师进修杂志

ObjectiveTo investigate the association of phosphorylation of mammalian target protein of rapamycin (pmTOR) expression with glioma malignancy grades,and the correlation of pmTOR expression with Survivin and Ki-67,which represent tumor cell anti-apoptosis ability and reproductive activity.MethodsImmunohistochemistry EliVision method was employed to detect the expression of pmTOR,Survivin and Ki-67 in paraffin tissues from 87 patients with glioma (grade Ⅰ - Ⅱ 27 cases,grade Ⅲ24 cases and grade Ⅳ 36 cases).The association between positive expression rate,level of pmTOR and malignancy grades,and the correlation of its expression level with Survivin and Ki-67 were further evaluated.Results There was no significant difference in the positive expression rate of pmTOR among grade Ⅰ - Ⅱ(77.8％,21/27),grade Ⅲ(75.0％,18/24) and grade Ⅳ (72.2％,23/36) (P ＞ 0.05).However,the significant association between pmTOR expression level and malignancy grades was observed.The expression from 87 patients with glioma was significantly positively correlated with Survivin and Ki-67 expression level (r =0.858,P ＜ 0.01 ;r =0.708,P ＜ 0.01 ).ConclusionsThe expression level of pmTOR is associated with malignancy grades,tumor cell anti-apoptosis ability and reproductive activity.pmTOR may be served as a useful marker for predicting the biological behavior of glioma and a useful target for gene therapy.