Downregulation of miRNA-127-5p aggravates spinal cord injury through activating MAPK1.

OBJECTIVE: To uncover the role of microRNA-127-5p (miRNA-127-5p) in aggravating motor dysfunction following spinal cord injury (SCI) by regulating the mitogen-activated protein kinase 1 (MAPK1) level. MATERIALS AND METHODS: In vivo SCI model in mice was established by constructing spinal cord hitting injury. Mice were classified into sham group, SCI group, SCI+miRNA-127-5p mimics group, and SCI+miRNA-127-5p mimics+MAPK1 group, respectively. Grip strengths of mouse pair forepaws, right forepaw, and left forepaw at different time points were determined. Expression levels of miRNA-127-5p and MAPK1 in mice of each group at post-SCI were detected. Potential binding sites in promoter regions of miRNA-127-5p and MAPK1 were predicted by bioinformatics and further confirmed by Dual-Luciferase reporter gene assay and Western blot. RESULTS: Grip strengths of SCI mice were much lower than those in sham group at different time points after SCI procedures. MiRNA-127-5p was markedly downregulated on the postoperative 3rd day in SCI group, and its level time-dependently decreased since after. In vivo overexpression of miRNA-127-5p in SCI mice improved their grip strengths from the postoperative 7th day. MAPK1 was the direct target of miRNA-127-5p. Transfection of miRNA-127-5p mimics downregulated protein level of MAPK1 in 293T cells. Overexpression of MAPK1 abolished the protective effect of miRNA-127-5p on motor function recovery following SCI. CONCLUSIONS: Downregulation of miRNA-127-5p aggravates SCI-induced motor dysfunction through negatively regulating MAPK1 level.

Expression Pattern of Tumor Necrosis Factor-α-Induced Protein 8-Like 2 in Acute Rejection of Cardiac Allograft.

BACKGROUND: Tumor necrosis factor-α-induced protein-8 like-2 (TIPE2) is a negative regulator of innate immunity and cellular immunity, yet the expression pattern of TIPE2 in acute rejection of cardiac allograft remain enigmatic. METHODS: We developed cardiac transplantation models and divided into 3 groups: a naive group, a syngeneic group, and an allogeneic group. Then, we detected the messenger RNA and protein of TIPE2 in cardiac allografts. Real-time polymerase chain reaction showed expression of CD4 and CD8 in the donor heart, and immunofluorescence assay revealed the association between T cells and TIPE2. RESULTS: In our study, we first found that the expression of TIPE2 in cardiac allografts is upregulated compared with the syngeneic control, and increases in a time-dependent manner. The immunocytochemistry of heart grafts revealed a strong expression of TIPE2 in the inflammatory cells, but not in the cardiomyocytes. Finally, we proved that CD4+ and CD8+ T cells infiltrated cardiac allografts abundantly, which express ample TIPE2. CONCLUSIONS: The upregulated expression of TIPE2 in cardiac allografts, mainly came from T cells, which infiltrated the donor heart. This finding indicates that there may be an association between TIPE2 and acute cardiac allograft rejection.

[Effect of Six1,TGF-ß,VEGF-C that promoting tumor lymphangiogenesis in human laryngeal carcinoma xenografts in nude mice].

Objective:To research the effect of Six1,TGF-ß,VEGF-C that promoting tumor lymphangiogenesis in human laryngeal carcinoma xenografts in nude mice. Method:Technology of RNA interference was used for silencing Six1 and TGF-ß genes expression in laryngeal squamous cell, preparation Six1-targeting and TGF-ß-targeting siRNA for transfecting into laryngeal squamous cell carcinoma, sieve out positive clone cell and amplify. Preparing bearing cancer mice, the mice were divided into five groups, group A (untransfected),group B(empty vector), group C(Six1-siRNA),group D(TGF-ß-siRNA),group E(Six1+TGF-ß-siRNA). 6-12mm when the tumor has grown, the mice were sacrificed by cervical. The size of each tumor and metastasis were observed and recorded.The protein expression of Six1,TGF-ß and VEGF-C was determined by immunohistochemistry and Western blot.The mRNA of Six1,TGF-ß and VEGF-C was determined by RT-PCR. Result:The average tumor volume and the number of metastasis cases in group B have no statistically significant compared with group A. The average tumor volume in group C, group D and group E has no significantly reduced, but there is a clear reduction of the number of metastasis cases compared with group A. The average tumor volume and number of metastasis cases in group E has no significantly reduced compared with group C and group D. Both protein and mRNA expression of Six1, TGF-ß and VEGF-C in group B had no significant difference compared with group A. In group C, group D and group E,both protein and mRNA expression of VEGF-C was decreased,difference has statistically significant compared with group A. Both protein and mRNA expression of VEGF-C in group E had no significant difference compared with group C and group D. Conclusion:Both Six1 and TGF-ß can mediate tumor lymphangiogenesis and lymph node metastasis by mediate the expression of VEGF-C. Suggest that Six1,TGF-ß might be a potential therapeutic target for preventing lymph node metastasis of tumor.

Dysregulation of Bmi1 promotes malignant transformation of hepatic progenitor cells.

Adult hepatic progenitor cells (HPCs) are involved in a wide range of human liver diseases, including hepatocellular carcinoma (HCC). Bmi1 has been reported to have vital roles in stem cell self-renewal and carcinogenesis. We have previously demonstrated that Bmi1 is upregulated in HCC with bile duct tumor thrombi, a subtype of HCC characterized by profuse expression of hepatic stem cell markers. However, the function of Bmi1 in HPCs has not yet been well elucidated. The current study was designed to investigate the effects of Bmi1 on the biological properties of rat HPCs. To accomplish this, Bmi1 was silenced or enhanced in two HPC cell lines (WB-F344 and OC3) by, respectively, using either small interfering RNA against Bmi1 or a forced Bmi1 expression retroviral vector. The biological functions of Bmi1 in HPCs were investigated through cell proliferation assays, colony-formation assays, cell cycle analysis and invasion assays, as well as through xenograft-formation assays. In this study, genetic depletion of Bmi1 repressed cell proliferation, colony formation and invasion in both assessed HPC cell lines relative to controls. Conversely, forced expression of Bmi1 in two HPCs cell lines promoted cell proliferation, colony formation and invasion in vitro. Aldehyde dehydrogenase (ALDH) assay revealed a significant increase in the number of ALDH-positive cells following the forced expression of Bmi1 in HPCs. Most importantly, transplantation of forced Bmi1 expression HPCs into nude mice resulted in the formation of tumors with histological features of poorly differentiated HCC. Taken together, our findings indicate that forced expression of Bmi1 promotes the malignant transformation of HPCs, suggesting Bmi1 might be a potential molecular target for the treatment of HCC.

[Strategy of the diagnosis and treatment for hydrocephalus associated with acoustic neuroma].

OBJECTIVE: To describe and analyze the strategy of the diagnosis and treatment for acoustic neuroma associated with hydrocephalus. METHODS: A retrospective review was performed in 29 patients with hydrocephalus associated with acoustic neuroma form Apr. 2004 to Apr. 2015. The patients' clinical information, the types of the hydrocephalus, the treatment and the prognosis of the hydrocephalus were recorded. RESULTS: There were 20 patients with obstructive hydrocephalus and 9 patients with communicating hydrocephalus preoperatively. Among the 29 cases, 3 patients had ventriculoperitoneal shunts, 5 patients had external ventricular drains, the remaining 21 patients had no further managements for hydrocephalus; after removing the acoustic neuroma, the hydrocephalus improved in 10 cases, the ventricle unchanged in 10 cases among the obstructive hydrocephalus group, the ventricle unchanged in all 9 cases among the communicating hydrocephalus group. Nineteen cases were diagnosed with communicating hydrocephalus and 10 cases with obstructive hydrocephalus postoperatively. CONCLUSIONS: Among the patients of acoustic neuroma associated with hydrocephalus, communicating hydrocephalus is more common than obstructive hydrocephalus. The optimal management of acoustic neuroma associated with hydrocephalus is complete removal of the tumor, with treatment only for patients with persistent hydrocephalus.

Expression of exogenous NF-IL6 induces apoptosis in Sp2/0-Ag14 myeloma cells.

NF-IL6 (or C/EBP beta) is a multifunctional transcription factor of the C/EBP family. To study the effects of exogenous expression of NF-IL6 on the growth of cells of hematopoietic tumors, we infected SP2/0-Ag14 myeloma cells with pLXSN recombinant retrovirus harboring the NF-IL6 cDNA. We found that the transfected cells revealed changes typical of the apoptosis and finally died. The expression of NF-IL6 protein was shown by a "supershift" assay in the NF-IL6-transfected cells. In controls, where cells were infected by the retroviral vector with a mutant NF-IL6 coding region as well as the vector without insert, no sign for expression of NF-IL6 and apoptosis was observed. Therefore, our work shows that the apoptosis of this myeloma cell line was induced by the exogenous NF-IL6 expression. This is significantly different from the apparent effects of NF-IL6 in leukemia cells and in fibroblasts. These facts indicate that NF-IL6 exerts different functions in different cell types. We discuss possible mechanisms for the NF-IL6-induced apoptosis.

[Changes of cytoplasmic calcium and magnesium concentration and calcium distribution in human platelets caused by thrombin].

Changes of calcium and magnesium concentration in human platelets during thrombin induced agglutination and the distribution of the respective concentration in the platelet population were measured by fluorescence probe using fura 2, mag-fura 2 and fluo 3. The cytoplasmic free calcium concentration increases with addition 2 thrombin. When the thrombin concentration was increased to 0.5 U/ml, the calcium concentration increase showed two rising peaks. The popular cytoplasmic calcium concentration in platelets shows normal distribution. The cytoplasmic free magnesium concentration also increases with increase of calcium concentration, showing that magnesium also plays a significant effect in the condensation of platelets.

Factors that can minimize bleeding complications after renal biopsy.

Renal biopsy is a very important diagnostic tool in the evaluation of renal diseases. However, bleeding remains to be one of the most serious complications in this procedure. Many new techniques have been improved to make it safer. The risk factors and predictors of bleeding after percutaneous renal biopsy have been extensively reported in many literatures, and generally speaking, the common risk factors for renal biopsy complications focus on hypertension, high serum creatinine, bleeding diatheses, amyloidosis, advanced age, gender and so on. Our primary purpose of this review is to summarize current measures in recent years literature aiming at minimizing the bleeding complication after the renal biopsy, including the drug application before and after renal biopsy, operation details in percutaneous renal biopsies, nursing and close monitoring after the biopsy and other kinds of biopsy methods.

Cleaning behaviors in four scorpion species

Scorpions rely predominantly on mechanosensory and chemosensory organs to guide their orientation behaviors. Once sensory organs are affected by the presence of dirt such as clay or prey bodily fluid, scorpions may display a cleaning behavior to reduce or eliminate its influence on their sensory capabilities. In the laboratory, cleaning behaviors of two buthid species, Mesobuthus eupeus (Koch, 1839) and Mesobuthus caucasicus (Nordmann, 1840), and one euscorpiid species, Scorpiops luridus Zhu Lourenço & Qi, 2005 from China, were observed before and after feeding. Moreover, two distinct cleaning behaviors in Scorpiops luridus and three in Heterometrus petersii (Thorell, 1876) (Scorpionidae) were noted for several times during daily activities. Based on these observations, we were able to conclude that different tools and the same tool with diverse applications are used for cleaning the same object in numerous scorpion species.(AU)

Courtship and mating of Scorpiops luridus Zhu Lourenço & Qi, 2005 (Scorpiones: Euscorpiidae) from Xizang province, China

In the current work, the courtship and mating of Scorpiops luridus Zhu Lourenço & Qi, 2005 (Euscorpiidae) from Xizang province (Tibet), China, were studied for the first time in the laboratory. Most of the mating behaviors in Scorpiops luridus are not remarkably different from those exhibited by other scorpions. However, for the first time a male pulling a female with its chelicerae to rapidly accomplish the sperm uptake was observed. Additionally, the sexual stinging behavior displayed by the male occurred in the initial stage, not during the promenade stage as previously described in several scorpion species. Through observation and analysis, we speculate that venom injection during sexual stinging is selective, possibly relying on the status shown by the stung scorpion (passive or aggressive). In order to clearly describe the process of courtship and mating, both sequences are represented in a flow chart, while the main behavior components of these processes were identified, analyzed and discussed.(AU)