Search

To evaluate the effect of interleukin-19 (IL-19) treatment on epidural fibrosis and its mechanism of action with transforming growth factor β (TGF-β).Initially, IL-19 (10, 20, 50 and 100 ng/L) was used to pretreat rat fibroblasts. TGF-β (10 μg/L) was then applied to activate fibroblasts. The protein expression levels of TGF-β receptor, extracellular-signal-regulated kinase (Erk) and p-38 were measured by Western blotting. In addition, we performed laminectomy at T10 vertebral plate in rats, followed by injection of IL-19 in caudal vein one week after injury. Furthermore, IL-19, TGF-β and fibrosis indexes were measured by quantitative Real-time polymerase chain reaction (qRT-PCR) and Western blotting at 7 and 28 days after injury, respectively.Concentration-dependent IL-19 significantly down-regulated TGF-β receptor expression and inhibited phosphorylated Erk (p-Erk) and phosphorylated p38 (p-p38). In vivo, IL-19 reduced the expressions of TGF-β and connective tissue growth factor (CTGF) at 7 days. Furthermore, IL-19 significantly suppressed extracellular matrix productions formation, including α smooth muscle actin (α-SMA) and collagen-1 (COL-1), and fibronectin at 28 days.IL-19 inhibited TGF-β expression via Erk and p38 pathway. Moreover, it decreased CTGF expression to suppress α-SMA, COL-1 and fibronectin in scar tissues, thereby preventing spinal cord from compression of scar tissues.

To investigate the regulatory role of microRNA-490-3p in the recovery process of spinal cord injury (SCI) and its underlying mechanism.Expression levels of microRNA-490-3p and MK2 in peripheral blood of SCI patients and healthy controls were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Glial cells C8-D1A and C8-B4 were induced by H2O2 for constructing in vitro SCI model, followed by detection of microRNA-490-3p and MK2 levels. After glial cells were transfected with microRNA-490-3p mimic or inhibitor, respectively, cell apoptosis and levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected. Bioinformatics was used to predict the binding site between microRNA-490-3p and MK2, which was further verified by dual-luciferase reporter gene assay. After construction of MK2 overexpression plasmid, rescue experiments were carried out to confirm the regulatory effect of microRNA-490-3p on MK2 in mediating SCI recovery.MicroRNA-490-3p expression was remarkably lower, whereas MK2 expression was higher in peripheral blood of SCI patients than those of healthy controls. Downregulated microRNA-490-3p and upregulated MK2 were observed in glial cells after H2O2 induction in C8-D1A and C8-B4 cells. Overexpression of microRNA-490-3p remarkably decreased levels of TNF-α and IL-6, as well as alleviated apoptosis in glial cells. Both protein and mRNA levels of MK2 were negatively regulated by microRNA-490-3p. Dual-luciferase reporter gene assay confirmed that MK2 was the target gene of microRNA-490-3p. Finally, rescue experiments verified that the regulatory effects of microRNA-490-3p on alleviating inflammation and apoptosis after SCI were reversed by MK2 overexpression.MicroRNA-490-3p is lowly expressed in glial cells after SCI. Overexpression of microRNA-490-3p alleviates inflammation and apoptosis via targeting MK2, thereafter promoting SCI recovery.

This study was conducted to explore the influencing factors of ova in vitro fertilization (IVF) and transfer of the fertilized ova into the oviduct of recipient hens. The efficiency of fertilization was compared using three aspects: (i) the different time of ova collection and transfer, (ii) egg-laying period of recipient hen; and (iii) semen volume. The following results are observed: 72%, 40% and 0% of ova were found in ovarian sac in 30∼40 min, 50∼60 min and more than 90 min post-oviposition, respectively; 20%, 18%, 14% and 5.8% of ova were fertilized with 0.1, 0.2, 0.5 and 1.0 ml semen, respectively; and 33% and 100% of healthy chickens were hatched from fertile ova with 0.1 and 0.5 ml of semen, respectively. All oocytes obtained from ovary and mid-oviduct were unfertilized. Embryos were transferred into recipient hens 30 min ± 10 min post-oviposition, and 70% of shelled eggs were produced. There were no eggs produced in the other transfer times. This demonstrated that live chicken can be obtained by IVF of ova collected shortly after oviposition. It was important that the ovum was transferred into the oviduct infundibulum of recipient hens immediately or shortly after oviposition.

Although many genetic alterations are known to be associated with human transitional cell carcinoma (TCC) of the urinary bladder, relatively little is known about the roles of these molecular defects, singular or in combination, in bladder tumorigenesis. We have developed a transgenic mouse model of bladder tumorigenesis using a 3.6-kb promoter of uroplakin II gene to drive the urotheliums-specific expression of oncogenes. In this study, we demonstrate that transgenic mice bearing a low copy number of SV40T transgene developed bladder carcinoma in situ (CIS), whereas those bearing high copies developed CIS as well as invasive and metastatic TCCs. These results indicate that the SV40T inactivation of p53 and retinoblastoma gene products, defects frequently found in human bladder CIS and invasive TCCs, can cause the aggressive form of TCC. Our results also provide experimental proof that CIS is a precursor of invasive TCCs, thus supporting the concept of two distinct pathways of bladder tumorigenesis (papillary versus CIS/invasive TCC). This transgenic system can be used for the systematic dissection of the roles of individual or combinations of specific molecular events in bladder tumorigenesis.

The MB-50 protocol worked out for long by the World Health Organization has been widely employed for the screening of male contraceptives. With this method, 40-80 rats will be used for 1 sample and the course of study will be about 3 months. In view of the high expenses and long duration of time needed, it seems to be not suitable for a preliminary screening. We have exploited a much simpler method, the intratesticular subcapsular assay. The method is as follows: Puncture the scrotal skin and the tunica albuginea at the equatorial plane of the testis just opposite to the epididymis with an intradermal needle fixed at a tuberculin syringe. Then push on the needle for 3-4 mm along the equator parallel to the tunica albuginea. Inject 100 microliters of solution containing 0.1-5 mg of the sample to each testis. To the controls, only the vehicle is injected. One week later the same procedure is repeated and 2 more weeks later, the animals are sacrificed and the data are observed (see Table 1 and 2). In this paper, 7 samples screened with this method were compared with results of the MB-50 method, no positive sample was missed except one false positive sample. These results indicate the coincidence between these two methods.