Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

The quality of life of organ transplant recipients is closely correlated with immune status. Compared with those undergoing other solid organ transplantation, the long-term prognosis of lung transplant recipients is worse. The underlying immune mechanism is complex with both similarities and characteristics. Therefore, in-depth understanding of the immune mechanism in the process of immune response of allogeneic lung transplantation plays a critical role in improving the long-term survival of the recipients. In this article, the unique composition of immune cells in the lung, the characteristics of rejection after lung transplantation, the early warning and differential diagnosis of pathogen infection in lung transplantation and postoperative complications after lung transplantation were investigated. Research progress on clinical diagnosis and basic studies related to immunology in allogeneic lung transplantation were summarized, aiming to elucidate the immunological characteristics of lung transplantation and provide theoretical basis for improving the longterm survival of lung transplant recipients and prevention and treatment of allograft dysfunction.

Objective To investigate the application value of Multi-Latex polygranular technique joint detection of kidney injury-related urinary microproteins in noninvasive diagnosis after renal transplantation. Methods Clinical data of 72 recipients undergoing renal transplantation were retrospectively analyzed. According to the level of serum creatinine (Scr), the recipients were divided into normal renal function group (group A, n=14), mild kidney injury (group B, n=37), and severe kidney injury group (group C, n=21). 20 healthy volunteers were selected as the healthy control group (HC group). The contents of urinary retinol binding protein (RBP), microalbumin (mAlb), IgG, transferrin (TRF), α₁-microglobulin (MG), and β₂-MG of subjects in each group were detected using the Multi-Latex polygranular technique. The correlation between urinary microproteins and Scr, blood urea nitrogen (BUN) was analyzed. The differences of urinary microproteins in each group were compared. And the diagnostic value of single and joint detection of urinary microproteins was evaluated. Results Six kinds of urinary microproteins in HC group and group A were significantly lower than those in group B and group C, and six kinds of urinary microproteins in group B were significantly lower than those in group C (all P < 0.01). Six kinds of urinary microproteins in renal transplant recipients were positively correlated with BUN. RBP, mAlb, α₁-MG, and β₂-MG were positively correlated with Scr. The correlations were statistically significant (P < 0.001-0.05). The diagnostic value of joint detection of urinary microproteins is better than the detection of single index, among which TRF+mAlb+RBP+α₁-MG quadruple detection had the highest diagnostic value. Conclusions Six kinds of urinary microproteins can be used as specific indicators to reflect graft renal function. The polygranular technique can simultaneously detect its contents and achieve noninvasive diagnosis. The diagnosis based on TRF+mAlb+RBP+α₁-MG quadruple detection is expected to further improve the noninvasive diagnosis system after renal transplantation.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

Objective To analyze and summarize the clinical features and diagnosis and treatment experience of brucellosis after renal transplantation. Methods Clinical data of one case with brucellosis after renal transplantation admitted to the 309thHospital of Chinese People's Liberation Army in October 2016 was collected. The clinical features, diagnosis and treatment were retrospectively analyzed. Clinical experience was summarized and literature review was conducted. Results At 3 months after renal transplantation, the patient suffered from temperature rise without known causes and presented with fever in the morning with a duration of 3 d. The route of infection was unknown, and the symptoms of alternative types of infection were not obvious. Empirical anti-infectious therapy was delivered for 1 week and yielded no efficacy. Blood culture test confirmed the diagnosis of brucella melitensis infection. The treatment included anti-infecting by the rifampicin, doxycycline, sulfamethoxazole, preventing the incidence of complications actively and protecting the liver and renal function. High clinical efficacy was achieved. During the 1-year follow up after discharge, the renal graft was stable and no other infectious symptoms, such as fever was found. Conclusions Brucellosis with unknown route of infection after renal transplantation is extremely rare and the common symptom is Malta fever. When the empirical anti-infectious treatment is not effective, blood culture and other related tests should be performed to confirm the diagnosis. The combination of rifampicin and doxycycline is recommended.

Objective:To establish the HPLC specific chromatogram and provide comprehensive evaluation of Xanthii fructus from different regions.Methods:The HPLC analysis was performed on an Alltima C18 column (250 mm ×4.6 mm,5μm) with acetonitrile-0.1％phosphoric acid solution as the mobile phase with gradient elution at a flow rate of 0.8 ml· min-1 .The detection wavelength was 278 nm.The column temperature was 25℃.The software"Similarity Evaluation System for Chromatographic Fingerprint of TCMs"was employed to carry out the similarity analysis of the samples from Henan , Jilin, Anhui and the other regions .Results:The specific chromatogram was preliminarily constructed and 5 common peaks with chlorogenic acid as the reference were identified .Conclusion:The method is scientific basis of the quality assessment of Xanthii fructus with convenient and reliable properties .

Objective To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.Methods The splenic CD8+T cells of Balb/c mice were obtained by CD8+f cell magnetic bead sorting kit.Under the effect of CD3/CD28-activated magnetic bead,CD8+T cells were stimulated by different concentrations of GABA.5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells.The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).Result Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells,manifested as significant decrease in the quantity of CD152+CD8+T cells.Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2,α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated.After CD8+T cell activation,the quantity of GABA-A receptor subtypes α3,αs,β2,β3,γ1,γ2 and θ were significantly increased,whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.Conclusions GABA can suppress the proliferation of activated CD8+T cells.The activation of CD8+T cells is regulated by GABA receptors.However,the underlying mechanism remains to be elucidated.

Objective To evaluate the effect of extracorporeal photopheresis upon the expression levels of interleukin (IL)-12p70 and T helper cell (Th) 1/Th2-like cytokines in splenic lymphocytes of mouse models undergoing skin transplantation. Methods The C57BL/6 mice were used as the donors and BALB/c mice as the recipients to establish mouse models with skin allograft. The splenic lymphocytes in the C57BL/6 and BALB/c mice (CSP and BSP) were isolated and treated with 8-methoxypsoralen combined long-wave ultraviolet (PUVA-SP). According to the components of intravenous infusion into the recipients, all experimental animals were randomly divided into the PUVA-BSP, PUVA-CSP, BSP, CSP and phosphate buffer solution (PBS) control groups (n=12 for each group). The mice were injected with PUVA-BSP, PUVA-CSP, BSP, CSP or PBS via the caudal vein at preoperative 7 d, upon the day of surgery and at postoperative 7 d, respectively. The apoptosis of the splenic lymphocytes was observed after PUVA treatment. The expression levels of IL-12p70 and Th1/Th2-like cytokines in the peripheral blood of the recipients were quantitatively measured. Results After the skin transplantation, the expression levels of IL-12p70 in the peripheral blood of mice in the PUVA-BSP and PUVA-CSP groups were significantly down-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). In the PUVA-BSP and PUVA-CSP groups, the expression levels of Th1-like cytokine IL-2, interferon (IFN)-γwere dramatically lower than those in the BSP, CSP and PBS control groups (all P<0.01). The expression levels of Th2-like cytokine IL-10 in the PUVA-BSP and PUVA-CSP groups were significantly up-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). Conclusions Infusion of PUVA-SP at a sufficient dose can induce the low expression level of IL-12p70 and drive the incidence of Th2 immune deviation in the recipient BALB/c mice.

Objective To investigate the influence of nerve growth factor (NGF) on the ability of differentiation and proliferation of regulatory T cells (Tregs) induced by mesenchymal stem cells (MSCs).Methods The MSCs were stimulated directly by NGF.IL-10,TGF-β and HLA-G were tested.The expression of CD4 and CD25 was detected by flow cytometry after co-culture.The expression of CD4,CD25 and Foxp3 was detected by flow cytometry after Transwell co-culture.Results As compared with control group,the expression of IL-10,TGF-β and HLA-G in NGF group was increased (P＜0.05 for all).The number of Tregs was increased after the co-culture (P＜0.05).The reduction in IL-10 and TGF-β could block the inducing function of NGF (P＜0.05).Conclusion NGF can enhance the ability of differentiation and proliferation of Tregs induced by MSC,which is possibly associated with the increases in the expression of IL-10 and TGF-β.

Objective To investigate the effect of infusion of spleen lymphocytes treated by extracorporeal photochemotherapy on the regulatory T cell (Treg)and survival time of skin allograft in mice. Methods The skin allograft model in mice was established with C57BL/6 mice as donors and BALB/c mice as recipients. The spleen lymphocytes (CSP,BSP)in mice C57BL/6 and BALB/c were isolated,and the mice spleen lymphocytes (PUVA-SP) treated with 8-methoxypsoralen plus ultraviolet (PUVA)were prepared. The experimental animals were randomly divided into 5 groups according to the compositions infused into the recipients through vein:PUVA-BSP,PUVA-CSP,BSP,CSP and phosphate buffer solution (PBS)control groups (n=12 in each group). All recipients of each group were injected with PUVA-BSP,PUVA-CSP,BSP,CSP or PBS on day 7 before the operation,on the operation day and day 7 after the operation through the tail vein,respectively. The survival time of graft in the recipients was observed,and the expression of CD4 +CD25 +Foxp3 +Treg in peripheral blood was detected. Results After skin allograft,the rate of CD4 +CD25 +Foxp3 +Treg in peripheral blood of the recipients in PUVA-BSP group and PUVA-CSP group was significantly higher than those of BSP, CSP and PBS control groups. The rate of CD4 +CD25 +Foxp3 +Treg in PUVA-CSP group was higher than that of PUVA-BSP group,while BSP and CSP groups were lower than that of PBS control group. The survival time of skin graft in the recipients in PUVA-BSP group and PUVA-CSP group was significantly longer than that of BSP,CSP and PBS control groups (all P<0.05 ). Conclusions Sufficient infusion of PUVA-SP can induce more CD4 +CD25 +Foxp3 +Treg in the recipients and prolong survival time of skin graft significantly.