Identification and analysis of the predictive urinary exosomal miR-195-5p in lupus nephritis based on renal miRNA-mRNA co-expression network.

OBJECTIVE: Lupus nephritis (LN) is the main complication of systemic lupus erythematosus (SLE), causing huge financial burden and poor quality of life. Due to the low compliance of renal biopsy, we aim to find a non-invasive biomarker of LN to optimize its predictive, preventive, and personalized medical service or management. METHOD: Herein, we provided a bioinformatic screen combined clinical validation strategy for rapidly mining exosomal miRNAs for LN diagnosis and management. We screened out differentially expressed miRNAs (DEMs) and differentially expressed mRNAs (DEGs) in LN database and performed a miRNA-mRNA integrated analysis to select out reliable changed miRNAs in LN tissues by using R and Cytoscape. Urinary exosomes were collected by ultracentrifugation and analyzed by nano-tracking analysis and western blotting. Detection of aquaporin-2 showed the tubular source of urinary exosomes. Urinary exosomal miRNAs were detected by RT-qPCR and the target of miR-195-5p was verified by using bioinformatic, dual-luciferase, and western blotting. RESULT: 15 miRNAs and their 60 target mRNAs were contained in miRNA-mRNA integrated map. Bioinformatic analysis showed these miRNAs were involved in various cellular biological process. Exosomal miR-195-5p, miR-25-3p, miR-429, and miR-218-5p were verified in a small clinical group (n = 47). Urinary exosomal miR-195-5p, miR-25-3p, and miR-429 were downregulated in patients and miR-195-5p could recognize LN patients from SLE with good sensitivity and specificity, showing good potential in LN disease monitoring and diagnosis. CONCLUSION: We analyzed and obtained a series of differential miRNAs in LN kidney tissues and suggested that urinary exosomal miR-195-5p could serve as a novel biomarker in LN. Further, miR-195-5p-CXCL10 axis could be a therapeutic target of LN.

Rapamycin preconditioning attenuates transient focal cerebral ischemia/reperfusion injury in mice.

Rapamycin, an mTOR inhibitor and immunosuppressive agent in clinic, has protective effects on traumatic brain injury and neurodegenerative diseases. But, its effects on transient focal ischemia/reperfusion disease are not very clear. In this study, we examined the effects of rapamycin preconditioning on mice treated with middle cerebral artery occlusion/reperfusion operation (MCAO/R). We found that the rapamycin preconditioning by intrahippocampal injection 20 hr before MCAO/R significantly improved the survival rate and longevity of mice. It also decreased the neurological deficit score, infracted areas and brain edema. In addition, rapamycin preconditioning decreased the production of NF-κB, TNF-α, and Bax, but not Bcl-2, an antiapoptotic protein in the ischemic area. From these results, we may conclude that rapamycin preconditioning attenuate transient focal cerebral ischemia/reperfusion injury and inhibits apoptosis induced by MCAO/R in mice.

Salidroside regulates imbalance of Th17/Treg and promotes ischemic tolerance by targeting STAT-3 in cerebral ischemia-reperfusion injury.

INTRODUCTION: The balance between Th17 and Treg cells controls the immune response and is an important regulator of helper T cells acting on autoimmune diseases. Focal cerebral ischemia-reperfusion injury can induce imbalance of Th17/Treg cells in the brain and the peripheral immune system of rats. The aim of this study was to investigate the effect of salidroside (Sal) on the ratio of Th17 and Treg cells in an adult rat model of middle cerebral artery occlusion (MCAO). MATERIAL AND METHODS: Forty rats were divided into 4 groups: normal group, sham group, surgery group, and Sal group. After treatment, the neurological deficits in rats were evaluated. Peripheral blood mononuclear cells were isolated and the count of Th17 and Treg cells was detected by flow cytometry. The infarct size and expression of RORγt and Foxp3 were detected in rat brain tissue. Rat spleen cells were isolated, CD4+ T cells were purified by immunomagnetic beads. Treg cells were induced by adding cytokine TGF-ß. Th17 cells were induced by adding cytokine IL-6. The expression of STAT-3 was inhibited by SiRNA, and the effect of Sal on the differentiation of Th17/Treg cells was analyzed. The expression levels of IL-6, TNF-α, MCP-1, STAT-3 and NF-κ-B2 proteins were examined. RESULTS: The results show that MCAO can induce an imbalance of Th17 and Treg cells in peripheral blood of rats. Sal treatment can significantly reduce the neurological deficit and infarct size of MCAO rats, reverse the oxidative stress of rat brain tissue, and inhibit the apoptosis of brain cells in MCAO rats. In the brain tissue of MCAO rats, Sal could significantly inhibit the expression of IL-6, TNF-α, MCP-1, STAT-3 and NF-κ-B2. Down-regulation of STAT-3 significantly reversed the therapeutic effects of Sal treatment. CONCLUSIONS: Our results indicate that Sal can increase the tolerance of rat brain tissue to ischemia, inhibit cell apoptosis and reduce oxidative stress by targeting STAT-3.

Correlation between MDM2 T309G single nucleotide polymorphism and esophageal cancer susceptibility: An updated meta-analysis.

BACKGROUND: The aim of this study was to investigate the correlation between MDM2 T309G single nucleotide polymorphism (SNP) and esophageal cancer susceptibility through pooling the open published data. METHODS: By systematic searching the databases of Medline, EMBASE, CBM and CNKI, the case-control or cohort studies related to MDM2 T309G single nucleotide polymorphism and esophageal cancer risk were screened. Genetic phenotype data of T309G single nucleotide was extracted from the original included studies. The correlation between MDM2 T309G single nucleotide polymorphism and esophageal cancer susceptibility was demonstrated by the odds ratio (OR) and its corresponding 95% confidence interval (95% CI). Publication bias was investigated by Egger's line regression test and begg's funnel plot. RESULTS: After systematic searching of the relevant database, nine publications were finally included in the present study. The combined data demonstrated that the subjects with the G genotype had an increased risk of developing esophageal cancer in dominant (OR = 1.13, 95% CI: 1.00-1.27, P = 0.043), recessive (OR = 1.27, 95% CI: 1.12-1.45, P = 0.000) and homozygous (OR = 1.34, 95% CI:1.04-1.74, P = 0.024) genetic model through random or fixed data pooling method. Both begg's and Egger's line regression test indicated no significant publication bias. CONCLUSION: Based on the present data, there was a significant correlation between MDM2 T309G single nucleotide polymorphism and esophageal cancer susceptibility. Individuals with G genotype may have an increased risk of developing esophageal cancer.

[Electromagnetic navigation interlocking intramedullary nail technology for treatment of femoral shaft fractures].

OBJECTIVE: To explore the value of electromagnetic navigation interlocking intramedullary nail in the treatment of femoral shaft fracture. METHODS: Between July 2012 and October 2013, 53 cases of femoral shaft fracture were treated. There were 40 males and 13 females, aged 16-52 years (mean, 38.3 years). The causes of injury were traffic accident in 28 cases, falling from height in 11 cases, falling in 7 cases, crush injury in 4 cases, and other in 3 cases. Of 53 cases, there were 3 cases of open fracture (Gustilo I degree) and 50 cases of closed fracture. Fracture was located in the proximal femur in 17 cases, middle femur in 29 cases, and distal femur in 7 cases. According to Winquist classification, 7 cases were rated as type I, 8 cases as type II, 22 cases as type III, and 16 cases as type IV; according to AO classification, 18 cases were rated as type 32-A, 28 cases as type 32-B, and 7 cases as type 32-C. The time from injury to operation was 3-11 days (mean, 5 days). Distal interlocking intramedullary nail was implanted using electromagnetic navigation. RESULTS: The distal locking nail operation with interlocking intramedullary nail was successfully completed under electromagnetic navigation; the one-time success rate of distal locking nail operation reached 100%; and the locking nail time was 5.0-9.5 minutes (mean, 7.0 minutes). Healing of incision by first intention was obtained after operation, and no complication of skin necrosis, infection, and sinus tract occurred. Fifty-three cases were all followed up 5-12 months (mean, 9 months). One case had hip pain and weaken middle gluteal muscle strength, and the symptoms disappeared after removing the nail. During the follow-up period, no broken nails, nail exit, infection, or re-fracture occurred. All fractures achieved clinical healing, and the healing time was 8-22 weeks (mean, 14.5 weeks). In 49 patients followed up 8 months, the Lysholm score was excellent in 44 cases, good in 4 cases, and acceptable in 1 case, with an excellent and good rate of 98%. CONCLUSION: Electromagnetic navigation system is safe and reliable, with the advantages of high positioning accuracy, short operation time, and no radiation, the clinical application of the system for distal locking nail operation can obtain excellent short-term effectiveness.

[Effects of salidroside on proliferation, apoptosis, phagocytosis, ROS and NO production of murine peritoneal macrophages in vitro].

AIM: To investigate the effects of salidroside(Sal) on proliferation, apoptosis, phagocytosis, the production of ROS and NO of murine peritoneal macrophages in vitro as well as its immunoregulation. METHODS: The single cell suspension of murine peritoneal macrophages was prepared under sterile condition, then co-cultured with different concentrations of Sal(80, 160 and 320 µmol/L)for 4 hours prior to stimulation with LPS and IFN-γ, the proliferation of macrophages was measured by MTT colorimetry. The effect of Sal on the apoptosis of Sytox® Green-labelled peritoneal macrophages induced by CHX was detected by Fluorescence enzyme-labelled meter. FCM was used to detect the effect of Sal on phagocytosis of peritoneal macrophages. Fluorescence enzyme-labelled meter was used to measure the effects of Sal on ROS of H(2);DCFDA-labelled macrophages induced by LPS and IFN-γ. Griess Gragent was used to detect the role of Sal in production of NO in peritoneal macrophages activated by LPS and IFN-γ. RESULTS: MTT result demonstrated that Sal could promote the proliferation of peritoneal macrophages activated by LPS and IFN-γ at the final concentrations of 80, 160, 320 µmol/L, respectively (P<0.05). The result of Fluorescence enzyme-labelled meter detected showed that Sal at the final concentration of 160 µmol/L could inhibit apoptosis of peritoneal macrophages induced by CHX(P<0.01). FCM analysis showed that different concentrations of Sal significantly promoted the phagocytosis of peritoneal macrophages which include un-activated and activated by LPS and IFN-γ(P<0.05). Fluorescence enzyme-labelled meter showed that Sal could reduce the production of ROS in activated peritoneal macrophages induced by LPS and IFN-γ(P<0.05). Sal also increased the production of NO in activated peritoneal macrophages induced by LPS and IFN-γ(P<0.05). CONCLUSION: Sal can promote proliferation of peritoneal macrophages stimulated by LPS and IFN-γ, and it can inhibit apoptosis of peritoneal macrophages induced by CHX, Sal also can promote the phagocytosis of peritoneal macrophages which include un-activated and activated by LPS and IFN-γ, Sal can reduce the production of ROS in activated peritoneal macrophages induced by LPS and IFN-γ, while Sal can promote the production of NO in activated peritoneal macrophages induced by LPS and IFN-γ.

[Effect of forsythia suspense extract on phagocytosis of peritoneal macrophages and NO production in vitro].

AIM: To investigate the effect of forsythia suspensa (FS) extract on phagocytosis of peritoneal macrophages and NO production in vitro. METHODS: The peritoneal macrophagess were isolated from BALB/c mice. After stained with CFDA-SE, the DH5alpha were co-cultured with peritoneal macrophagess for 3 h. The effect of FS extract on cyto-phagocytesis in vitro was analyzed by flow cytometry. The peritoneal macrophages were stimulated and activated by LPS in vitro. The effect of FS extract on NO production of the peritoneal macrophages in vitro was measured by NO assay kit. RESULTS: FCM analysis showed that FS extract significantly promoted the phagocytosis of peritoneal macrophages at the final concentration of 40, 80, 160 mg/L, respectively (P<0.05). It also decreased the production of NO at different concentration induced by LPS (P<0.05). CONCLUSION: FS extract can promote phagocytosis of peritoneal macrophages and inhibit NO production in vitro.

[Effects of Forsythia suspense extract on cell proliferation and activation of the mouse T lymphocytes in vitro].

AIM: To investigate the effect of Forsythia suspensa (FS) extract on the activation and proliferation of the mouse T lymphocytes in vitro as well as its immunosuppressive effect. METHODS: The lymphocytes were isolated from the lymphoid nodes of BALB/c mice. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69, CD25 and CD 71 of the activated T lymphocytes in vitro in response to Concanavalin A (ConA). Stained with CFDA-SE, the mouse lymphocytes were stimulated by ConA. The effect of FS extract on cell proliferation in vitro was analyzed by flow cytometry. RESULTS: FS extract decreased the expression of CD69, CD25 and CD71 at the final concentration of 40, 80 and 160 mg/L, respectively(P<0.05). CFDA-SE staining showed that FS extract at different concentration significantly inhibited the proliferation of lymphocytes induced by ConA (P<0.05). CONCLUSION: FS extract can inhibit the activation and proliferation of T lymphocytes in response to ConA.