With the hereotopic heart transplantation, immune tolerance induced by portal venous inoculation of donor spleen cells was studied.Methods:The recipient rats received donor spleen cells through portal vein combined with of cyclosporine A(CsA).The NK cell activity and IL-2, IFN-y expression of recipient spleen cells were detected. Results: Hie inoculation of donor spleen cells through the portal vein could significandy prolong survival time of heart allografts.IL- 2, IFN-y expression of recipient spleen lymphocytes and the recipient NK cell activity was also inhibitied.Conclusion:The inoculation of donor spleen cells through the portal vein could induce immune tolerance.The suppression of IL-2-NK-IFN-y immunologic net may be an important mechanism of portal vein tolerance.

Objective To investigate the change of cognitive function and cerebral blood flow in patients with transient ischemic attack(TIA).Methods 35 patients with TIA and 33 normal controls who matched in sex, age, right handed and education were tested by events related potential(ERP), the scale of elderly cognitive function(SECF) and magnetic resonance angiography(MRA).Results The peak latencies of P 3 components of ERP in the patients were significantly delayed as compared with the control group( P

OBJECTIVETo assess the quality of reporting by randomized controlled trial (RCT) related to dental implants in China during 2000 to 2012 by using the revised Jadad scale and consolidated standards of reporting trials (CONSORT) (2010) statement.

METHODSThe following electronic databases were searched: Chinese Biomedical Literature Database, Database for Chinese Technical Periodicals, China National Knowledge Infrastructure, PubMed, and EMBASE. A total of 19 journals of stomatology in China were also searched manually. The qualities of RCT with dental implant published between 2000 and 2012 were assessed using CONSORT (2010) statement and revised Jadad scale.

RESULTSTwenty-eight RCTs related to dental implants were identified. The quality of reporting in 28 articles was low. The mean revised Jadad score was 1.29 ± 0.71 and the CONSORT (2010) score was 9.75 ± 3.60.

CONCLUSIONThe methodological qualities of the included studies on dental implants are generally low, and reporting quality remain unsatisfactory.

China ; Dental Implants ; Humans ; Publishing ; Randomized Controlled Trials as Topic

Objective To evaluate the alternate intravesical instillation of MMC and IL 2 together with BCG for the prevention of bladder cancer recurrence. Methods The instillation was carried out for 41 cases of bladder cancer after surgical management.All the patients have been followed up for 38～66 months with an average of 50 months.TNFa levels were periodically detected after the instillation. Results No recurrence has been noted in all the 34 patients of Ta,T 1 and T 2 tumors except 1 with mixed squamous cancer. 3 of 7 T 3 cases underwent TURBT,recurrence was noted in 2 and the TNFa remained much higher after the instillation. Conclusions Alternate intravesical instillation of MMC and IL 2 together with BCG is an effective means of preventing bladder cancer recurrence.TNFa detection might serve as a marker for bladder cancer recurrence.

Objective:To investigate the receptor pathways of glycated basic fibroblast growth factor (bFGF) on proliferation and vascularization of human dermal microvascular endothelial cells (HDMECs).Methods:The experimental research method was used. Glycated bFGF stimulating solution was prepared with glucose and bFGF. Cells of the third to sixth passages were subcultured from the HDMEC primary cell line. The cells were divided into normal control group, glycated bFGF alone group, small interfering RNAs (siRNA)-advanced glycation end product receptor (RAGE) alone group, and siRNA-RAGE+glycated bFGF group, and seeded into 96-well plate with 6 wells in each group, and seeded into 6-well plate with 3 wells in each group. Cells in siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group were transfected with siRNA-RAGE and then were added into HDMEC culture medium for routine culture. After two days, the original HDMEC culture medium was discarded, and cells in siRNA-RAGE alone group were routinely cultured in HDMEC culture medium, cells in siRNA-RAGE+glycated bFGF group were routinely cultured in glycated bFGF stimulating solution. Cells in normal control group were routinely cultured in HDMEC culture medium, and cells in glycated bFGF alone group were routinely cultured in glycated bFGF stimulating solution. After transfection with siRNA-RAGE, cells were seeded into 48-well plate and divided into siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group, with 4 wells in each group. Another cells were directly seeded into 48-well plate without transfection and divided into normal control group and glycated bFGF alone group, with 4 wells in each group. Cells in normal control group and siRNA-RAGE group were routinely cultured in HDMEC culture medium. Cells in siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group were routinely cultured in glycated bFGF stimulating solution. Cells were divided into normal control group, glycated bFGF alone group, siRNA-fibroblast growth factor receptor (FGFR) alone group, and siRNA-FGFR+glycated bFGF group and seeded into 96-, 6- and 48-well plates. respectively. Cell density, sample number, and corresponding treatment were the same as before. Only siRNA-RAGE was replaced by siRNA-FGFR. Cells were divided into siRNA-positive control group, siRNA-negative control group, siRNA-RAGE group, and siRNA-FGFR group and transfected with siRNA-glyceraldehyde-3-phosphate dehydrogenase positive control, siRNA-negative control, siRNA-RAGE, and siRNA-FGFR for 4 to 6 hours, and then were added into HDMEC culture medium for routine culture. The transfection effect of siRNA was identified by reverse transcription PCR. Cell counting kit 8 method was used to determine the proliferation of cells after 2 days of culture (sample number was 6), flow cytometry was used to detect the apoptosis of cells after 2 days of culture (sample number was 3), tube forming test was used to detect the angiogenesis of cells cultured after 6 hours of culture (sample number was 4). Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results:At the 200 bp band, there were no complementary DNA (cDNA) bands in siRNA-positive control group, siRNA-RAGE group, and siRNA-FGFR group, but cDNA bands were detected in siRNA-negative control group, indicating the success of siRNA transfection. After 2 days of culture, the absorbance value of cells in glycated bFGF alone group was significantly lower than that of normal control group ( t=2.359, P<0.05); absorbance value of cells in siRNA-RAGE+glycated bFGF group was significantly higher than that of glycated bFGF alone group ( t=3.858, P<0.01), which was similar to that of siRNA-RAGE alone group ( t=2.148, P>0.05). The absorbance value of cells in siRNA-FGFR+glycated bFGF group was similar to that of glycated bFGF alone group ( t=0.805, P>0.05), but significantly lower than that of siRNA-FGFR alone group ( t=4.201, P<0.01). After 2 days of culture, the apoptotic rate of cells in glycated bFGF alone group was significantly higher than that of normal control group ( t=2.416, P<0.05). The apoptotic rate of cells in siRNA-RAGE+glycated bFGF group was significantly lower than the rates in glycated bFGF alone group and siRNA-RAGE alone group ( t=3.861, 2.724, P<0.05 or P<0.01). There were no significant differences in apoptosis rate of cells among normal control group, glycated bFGF alone group, siRNA-FGFR alone group, and siRNA-FGFR+glycated bFGF group ( F=2.218, P>0.05). After 6 hours of culture, the number of tubules of cells in normal control group (636±5) was significantly more than that of glycated bFGF alone group [580±8, t=10.825, P<0.01], and the number of tubules of cells in siRNA-RAGE+glycated bFGF group (647±10) was significantly more than those of glycated bFGF alone group and siRNA-RAGE alone group (628±4, t=13.040, 3.641, P<0.01). After 6 hours of culture, the number of tubules of cells in siRNA-FGFR+glycated bFGF group (619±5) was significantly more than that of glycated bFGF alone group ( t=9.000, P<0.01), but less than that of siRNA-FGFR alone group (632±3, t=2.814, P<0.05). Conclusions:Glycated bFGF affects the angiogenesis of HDMEC through RAGE pathway, which may be one of the reasons for the impaired wound healing of diabetic skin.

BACKGROUND: Researches suggest that transient ischemic attack (TIA)can induce cognitive dysfunction, and cerebral blood flow and its distribution are hypothesized to be closely related to cognitive activities.OBJECTIVE: To investigate the alteration of cognitive function and provide insights into its relations with cerebral perfusion in TIA patients.DESIGN: A case-control study.SETTING: Departments of Geriatrics, Electrophysiology and Magnetic Resonance of Urumqi General Hospital of Lanzhou Military Area Command of Chinese PLA.PARTICIPANTS: Totally 35 male right-handed TIA patients aged 45-78 years with an average of (68.1±8.4) years were selected from the inpatients and outpatients in the Department of Geriatrics, Urumqi General Hospital of Lanzhou Command of Chinese PLA between January 2002 and June 2003. Another 33 healthy right handed male subjects aged 45-77 years with an average of (67.8±8.6) years coming for physical examination were recruited to serve as the control group.METHODS: Patients and control subjects were tested with event-related potentials (ERPs) and the scale of elderly cognitive function (SECF) to examine the orientation, learning and memory, span, recall 1 (association),long-term memory, naming of animals, calculation, classification, copying,language and recall 2 (relation). According to the T score transformation table, the original scores were transformed into T scores relative to the age to eliminate the impact of age, and also into T'score to eliminate the interference by the patients'education, so that cognitive function of the patients could be evaluated with T'score, and the lower the score, the poorer the cognitive function. Cases in the two groups were all tested, and TIA patients were also examined with magnetic resonance angiography (MRA).MAIN OUTCOME MEASURES: Results of ERPs, SECF and MRA.RESULTS: Of the 35 TIA patients and 33 control subjects all completed the trial. Examination of ERPs reveled significantly prolonged latency of P300 components of ERP in the TIA group [(336.2±34.2) ms] than that in the control group [(311.3±44.2) ms, P ＜ 0.05]. The scores of span, recall 1,long-term memory, naming of animals, calculation, and recall 2 in SECF in TIA group were all lower than those in control group (39.7±11.9 vs 47.4±12.0; 54.5±14.8 vs 61.8±14.5; 61.1±7.8 vs 64.7±1.7; 59.4±11.0 vs 64.7±8.8; 50.0±14.7 vs 58.1±14.2; 44.6±15.4 vs 53.2±17.8, t=4.151 0-7.292 8, P ＜ 0.05-0.01). MRA identified abnormalities in 33 of the 35 TIA patients (94%), manifested mainly by stenosis and occlusion involving the vertebral artery (54%, 19/35), bilateral anterior, middle and posterior cerebral arteries (40% ,28/70;59% ,41/70;47% ,33/70), basilar artery (5.71%, 2/35) and bilateral internal carotid artery (5.71%, 4/70) respectively.CONCLUSION: TIA patients are characterized by prolonged P300 latency with multiple cognitive impairments especially in memory and cerebral artery stenosis and occlusion as shown by MRA, suggests that TIA patients have persistent low cerebral perfusion and frequently, cognitive dysfunction in the presence of local blood supply disorder in the hemispheres.

Objective To identify the cross-reactive antigens shared by Mycobacteria smegmatis(MS) and Mycobacteria tuberculosis(MTB) and to analyze their antigenicity.Methods Bacterial antigens were extracted from strains of MS and MTB by ultrasonication.Western blot assay was performed to analyze common antigens that reacted with both of the antiserum samples against MS and MTB.The extracted bacterial antigens were mixed with incomplete Freund′s adjuvant and then were injected into muscles of mice.Cytokines secreted by murine spleen lymphocytes following stimulation with various antigens of MS and MTB were determined by ELISPOT and flow cytometry on the 7th day.IgG levels in serum samples were detected by ELISA 7 days after injection.Results There were cross-reactive antigens shared by MS and MTB.Potent humoral immune responses and cellular immunity against both MS and MTB could be induced by those cross-reactive antigens after sensitization the mice by either MS or MTB antigens.Cytokines of IL-2 and IFN-γ in CD4+ and CD8+T cells of mice stimulated with MS or MTB antigens were significantly increased as compared with those of non-sensitization group and those of Brucella antigens stimulation group.ConclusionCross-reactive antigens shared by MS and MTS can effectively promote specific immune reactions to the infection of MTB, which provides a scientific basis for the development of tuberculosis vaccines.

Objective To explore the effect of diabetes mellitus (DM) on biological behavior of epidermal keratinocyte in rats. Methods A total of 40 Sprague-Dawley rats were randomized into control group and streptozotocin (STZ) -induced diabetes group. Of each group, 10 rats were implemented with deep partial-thickness scalding. The re-epithelialization rate was observed at the 3rd, 7th, 14th and 21th post-burn day. Histological characteristics and thickness of epidermal tissue in both groups were observed. The adhesion rate, cell cycles, apoptosis rate and migration ability of keratinocyte were measured. The accumulation of advanced glycosylation end products (AGEs) of epidermal tissue was observed. Results The percentages of re-epithelialized area at the 7th, 14th and 21th post-burn day were much lower in DM group than in control group (P<0.05). In DM group, the epidermal thickness was reduced obviously with obscure multilayered epithelium and less amount of prickle cells; The adhesion rates of 12, 24 h after culturing keratinocyte and the percentage of G2/M phase cells were lower in DM group than in control group (P<0.05). However, apoptosis rate of keratinocyte was higher, the amount of migration cell was significantly less in DM group than in control group (both P<0.05). There were lots of AGEs accumulated in epidermal tissue in DM group, while there were hardly AGEs in control group. Conclusions Re-epithelization blocked exists on non-healing wound in DM rats, which is related with the impaired keratinocyte biological behavior. A large of AGEs accumulate in the epidermal tissue of DM rats, which may be a important reason to inhibit keratinocyte function in diabetic environment.

OBJECTIVETo investigate the mechanism of immune escape in renal cell carcinoma(RCC).

METHODSFas and FasL expressions were examined by immunohistochemical technique in 44 RCC patients, with the Ki67 expression and apoptosis of tumor infiltrating lymphocytes(TIL) monitored simultaneously. Cytokines including IL2 and IFN alpha were used to induce the expression of the renal carcinoma cell lines 786-0 cells. Combination treatment of 786-0 with cytokines and Anti-Fas monoclonal antibody (FasAb) was used to induce apoptosis. FasL function was assessed by in vitro co-culture assays using renal cancer cells 786-0 and Fas-sensitive Jurkat T-cells.

RESULTS(1) Fas expression rate in RCC(22.8%) was lower than that in the controlled normal kidney tissues(53.8%, P < 0.01). FasL expression rate in RCC (46.5%) was higher than that in the controlled normal kidney tissues (23.2%, P < 0.01). That of Ki67 was 32.8%, with the expressions of Fas and Ki67 showing a negative correlation (r = -0.62, P < 0.05). In contrast, the expressions of FasL and Ki67 showed a positive correlation. (r = 0.93, P < 0.01). The Fas expression of stage I was significantly higher than that of stages III and IV. The expression rate of FasL in RCC was significantly increased with RCC stage (P < 0.01). (2) The apoptotic rate of TIL in RCC (33.9%) was significantly higher than that of the normal kidney tissues (3.5%, P < 0.01). The expression of FasL and the apoptotic rate of TIL in RCC gave a positive correlation (r = 0.96, P < 0.01). (3) Fas expression rate in 786-0 cells was 13.7%. The apoptotic rate mediated by FAsAb was 9.6%. IFN alpha was able to up-regulate the Fas expression and subsequently augment the FasAb-mediated apoptosis in 786-0 cells. But IL2 did not show similar effects. (4) The FasL expression rate of 786-0 was 18.6%. FasL expressed by 786-0 cells was able to induce apoptosis of Jurkat T-cells in co-culture assays and the apoptosis of Jurkat T-cells was significantly lowered after blocking the effect of FasL with Fas-neutralizing antibody NOK-2, giving the apoptotic rates of 14.9% and 2.0%, respectively, the difference therein is statistically significant (P < 0.01).

CONCLUSIONDown-regulation of Fas expression and up-regulation of FasL-expression are the mechanisms through which the RCC cells escape from immune attack.

Adult ; Aged ; Carcinoma, Renal Cell ; immunology ; Fas Ligand Protein ; Female ; Humans ; Ki-67 Antigen ; immunology ; Kidney Neoplasms ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Middle Aged ; fas Receptor ; immunology

Objective:To investigate the receptor pathways of glycated basic fibroblast growth factor (bFGF) on proliferation and vascularization of human dermal microvascular endothelial cells (HDMECs).Methods:The experimental research method was used. Glycated bFGF stimulating solution was prepared with glucose and bFGF. HDMECs of the third to sixth passages were used in the experiment. Cells were divided into small interfering RNA (siRNA)-positive control group, siRNA-negative control group, siRNA-receptor for advanced glycation end product (RAGE) group, and siRNA-receptor for fibroblast growth factor (FGFR) group and transfected with siRNA-positive control glyceraldehyde-3-phosphate dehydrogenase, siRNA-negative control, siRNA-RAGE, and siRNA-FGFR for 4 to 6 hours, and then were added into HDMEC culture medium for routine culture. The transfection effect of siRNA was identified by reverse transcription polymerase chain reaction. The cells were divided into normal control group, glycated bFGF alone group, siRNA-RAGE alone group, and siRNA-RAGE+ glycated bFGF group, and seeded into 96-well plate and 6-well plate. Cells in siRNA-RAGE alone group and siRNA-RAGE+ glycated bFGF group were transfected with siRNA-RAGE and then were added into HDMEC culture medium for routine culture. After two days, the original HDMEC culture medium was discarded, and cells in siRNA-RAGE alone group were routinely cultured in HDMEC culture medium, cells in siRNA-RAGE+ glycated bFGF group were routinely cultured in glycated bFGF stimulating solution. Cells in normal control group were routinely cultured in HDMEC culture medium, and cells in glycated bFGF alone group were routinely cultured in glycated bFGF stimulating solution. After transfection with siRNA-RAGE, cells were seeded into 48-well plate and divided into siRNA-RAGE alone group and siRNA-RAGE+ glycated bFGF group. Another cells were directly seeded into 48-well plate without transfection and divided into normal control group and glycated bFGF alone group. Cells in the 4 groups were conducted with the corresponding treatment as above. Cells were divided into normal control group, glycated bFGF alone group, siRNA-FGFR alone group, and siRNA-FGFR+ glycated bFGF group and seeded into 96-, 6-, and 48-well plates, respectively, with the corresponding treatment the same as above. Only siRNA-RAGE was replaced by siRNA-FGFR. Cell counting kit 8 method was used to determine the proliferation of cells after 2 days of culture (sample number was 6), flow cytometry was used to detect the apoptosis of cells after 2 days of culture (sample number was 3), tube forming test was used to detect the angiogenesis of cells after 6 hours of culture (sample number was 4). Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results:At the 200 bp band, there were no target genes in siRNA-positive control group, siRNA-RAGE group, or siRNA-FGFR group, but target genes were detected in siRNA-negative control group, indicating the success of siRNA transfection. After 2 days of culture, the absorbance value of cells in glycated bFGF alone group was significantly lower than that of normal control group ( t=2.359, P<0.05); absorbance value of cells in siRNA-RAGE+ glycated bFGF group was significantly higher than that of glycated bFGF alone group ( t=3.858, P<0.01), which was similar to that of siRNA-RAGE alone group ( t=2.148, P>0.05). The absorbance value of cells in siRNA-FGFR+ glycated bFGF group was similar to that of glycated bFGF alone group ( t=0.805, P>0.05), but significantly lower than that of siRNA-FGFR alone group ( t=4.201, P<0.01). After 2 days of culture, the apoptotic rate of cells in glycated bFGF alone group was significantly higher than that of normal control group ( t=2.416, P<0.05). The apoptotic rate of cells in siRNA-RAGE+ glycated bFGF group was significantly lower than the rates in glycated bFGF alone group and siRNA-RAGE alone group ( t=3.861, 2.724, P<0.05 or P<0.01). There were no statistically significant differences in apoptosis rate of cells among normal control group, glycated bFGF alone group, siRNA-FGFR alone group, and siRNA-FGFR+ glycated bFGF group ( F=2.218, P>0.05). After 6 hours of culture, the number of tubules of cells in normal control group (636±5) was significantly more than that of glycated bFGF alone group (580±8, t=10.825, P<0.01), and the number of tubules of cells in siRNA-RAGE+ glycated bFGF group (647±10) was significantly more than those of glycated bFGF alone group and siRNA-RAGE alone group (628±4, t=13.040, 3.641, P<0.01). After 6 hours of culture, the number of tubules of cells in siRNA-FGFR+ glycated bFGF group (619±5) was more than that of glycated bFGF alone group ( t=9.000, P<0.01), but less than that of siRNA-FGFR alone group (632±3, t=2.814, P<0.05). Conclusions:Glycated bFGF affects the proliferation and angiogenesis of HDMEC through RAGE pathway, which may be one of the reasons for impaired wound healing of diabetic skin.