[Trametenolic acid inhibits migration and invasion of hepatocellular carcinoma HepG2.2.15 cells via RhoC/ROCK1 pathway].

This study investigated the effect of trametenolic acid(TA) on the migration and invasion of human hepatocellular carcinoma HepG2.2.15 cells by using Ras homolog gene family member C(RhoC) as the target and probed into the mechanism, aiming to provide a basis for the utilization of TA. The methyl thiazolyl tetrazolium(MTT) assay was employed to examine the proliferation of HepG2.2.15 cells exposed to TA, and scratch and Transwell assays to examine the cell migration and invasion. The pull down assay was employed to determine the impact of TA on RhoC GTPase activity. Western blot was employed to measure the effect of TA on the transport of RhoC from cytoplasm to cell membrane and the expression of RhoC/Rho-associated kinase 1(ROCK1)/myosin light chain(MLC)/matrix metalloprotease 2(MMP2)/MMP9 pathway-related proteins. RhoC was over-expressed by transient transfection of pcDNA3.1-RhoC. The changes of F-actin in the cytoskeleton were detected by Laser confocal microscopy. In addition, the changes of cell migration and invasion, expression of proteins in the RhoC/ROCK1/MLC/MMP2/MMP9 pathway, and RhoC GTPase activity were detected. The subcutaneously transplanted tumor model of BALB/c nude mice and the low-, medium-, and high-dose(40, 80, and 120 mg·kg~(-1), respectively) TA groups were established and sorafenib(20 mg·kg~(-1)) was used as the positive control. The tumor volume and weight in each group were measured, and the expression of related proteins in the tumor tissue was determined by Western blot. The results showed that TA inhibited the proliferation of HepG2.2.15 cells in a concentration-dependent manner, with the IC_(50) of 66.65 and 23.09 µmol·L~(-1) at the time points of 24 and 48 h, respectively. The drug administration groups had small tumors with low mass. The tumor inhibition rates of sorafenib and low-, medium-and high-dose TA were 62.23%, 26.48%, 55.45%, and 62.36%, respectively. TA reduced migrating and invading cells and inhibited RhoC protein expression and RhoC GTPase activity in a concentration-dependent manner, dramatically reducing RhoC and membrane-bound RhoC GTPase. The expression of ROCK1, MLC, p-MLC, MMP2, and MMP9 downstream of RhoC can be significantly inhibited by TA, as confirmed in both in vitro and in vivo experiments. After HepG2.2.15 cells were transfected with pcDNA3.1-RhoC to overexpress RhoC, TA down-regulated the protein levels of RhoC, ROCK1, MLC, p-MLC, MMP2, and MMP9 and decreased the activity of RhoC GTPase, with the inhibition level comparable to that before overexpression. In summary, TA can inhibit the migration and invasion of HepG2.2.15 cells. It can inhibit the RhoC/ROCK1/MLC/MMP2/MMP9 signaling pathway by suppressing RhoC GTPase activity and down-regulating RhoC expression. This study provides a new idea for the development of autophagy modulators targeting HSP90α to block the proliferation and inhibit the invasion and migration of hepatocellular carcinoma cells via multiple targets of active components in traditional Chinese medicines.

FALEC exerts oncogenic properties to regulate cell proliferation and cell-cycle in endometrial cancer.

Focally amplified lncRNA on chromosome 1 (FALEC) is novel lncRNA located in a focal amplicon on chromosome 1q21.2, and has been identified as an oncogenic properties in a variety of human cancers. However, there was no report about the expression pattern and biological function of FALEC in endometrial cancer. In our research, FALEC expression was increased in endometrial cancer tissue samples and cell lines compared with corresponding paracancerous normal tissue samples and cell line, respectively. Furthermore, we investigated the clinical significance of FALEC in endometrial cancer patients, and found endometrial cancer patients with advanced clinical stage or large tumor size had higher levels of FALEC expression than those with early clinical stage or small tumor size. The in vitro studies showed silencing of FALEC expression inhibited cell proliferation and arrested cell cycle at G0/G1. In conclusion, FALEC is overexpressed in endometrial cancer tissues and cells, and involved in regulating cell proliferation and cell-cycle.

Long-term effects of oral tea polyphenols and Lactobacillus brevis M8 on biochemical parameters, digestive enzymes, and cytokines expression in broilers.

This study investigates the long-term effects of oral tea polyphenols (TPs) and Lactobacillus brevis M8 (LB) on biochemical parameters, digestive enzymes, and cytokines expression in broilers. In experiment 1, 240 broiler chickens were selected to investigate the effects of 0.06 g/kg body weight (BW) TP and 1.0 ml/kg BW LB on broilers; in experiment 2, 180 broiler chickens were assigned randomly to three groups to investigate the effects of different dosages of TP (0.03, 0.06, and 0.09 g/kg BW) combined with 1.0 ml/kg BW LB on broilers; in experiment 3, 180 broiler chickens were assigned randomly to three groups to investigate the effects of different dosages of LB (0.5, 1.0, and 1.5 ml/kg BW) combined with 0.06 g/kg BW TP on broilers. The results showed that TP and LB affected serum biochemical parameters, and TP reduced serum cholesterol (CHO) and low-density lipoprotein cholesterol (LDL-C) abundances in a dosage-dependent manner (P<0.05) on Day 84. Meanwhile, broilers fed a diet supplemented with TP or LB had a lower intestinal lipase activity on Day 84 compared with the control group (P<0.05). Middle and high dosages of TP increased pancreatic lipase and proventriculus pepsin activities (P<0.05). Also middle and high dosages of LB significantly enhanced pancreatic lipase activity (P<0.05), while high LB supplementation inhibited intestinal trypsase (P<0.05) on Day 84. Furthermore, both TP and LB reduced intestinal cytokine expression and nuclear factor-κ B (NF-κB) mRNA level on Days 56 and 84. In conclusion, long-term treatment of TP and LB improved lipid metabolism and digestive enzymes activities, and affected intestinal inflammatory status, which may be associated with the NF-κB signal.

Magnetic resonance imaging of ruptured spinal dermoid tumors with spread of fatty droplets in the central spinal canal and/or spinal subarachnoidal space.

OBJECTIVE: To evaluate magnetic resonance imaging (MRI) features of ruptured spinal dermoid tumors with spread of lipid droplets in the central spinal canal and/or spinal subarachnoid space and to understand the underlying mechanism. METHODS: The MRI features of 12-ruptured spinal dermoid tumors were retrospectively analyzed. A literature review was performed to analyze the reported cases of ruptured spinal dermoid tumors along with our cases. RESULTS: The locations of dermoids in our series are all at or below T12 level. Of the 12 cases, 10 ruptured into the central spinal canal, 1 ruptured into the central spinal canal as well as the subarachnoid space, and 1 ruptured into subarachnoid space only. Free lipid droplets exhibited hyperintensity on T1 weighted images, hypointensity on T2 weighted images, and low signal on fat-suppression sequence. CONCLUSION: Spinal dermoid tumors ruptured into central spinal canal and/or spinal subarachnoid space have unique MRI features. The absorption of lipid droplets within central spinal canal is rather difficult, and their movement is extremely slow. We propose that fatty components within the central canal of spinal cord may be partially associated with spinal dermoid tumors developmentally.

Characteristic analysis on susceptibility weighted imaging of intravitreous foreign body of autologous eyelashes in rabbits.

OBJECTIVE: To explore the characteristics of susceptibility weighted imaging (SWI) of the intravitreous foreign body of autologous eyelashes in rabbits. METHODS: A total of 12 New Zealand white rabbits, either sex, weighing 2.5-3.5 kg, and provided by the Experimental Animal Center of Henan Province were employed in this study. For each rabbit, 5 autologous eyelashes (1 cm in length and 0.2-0.3 mm in diameter) were implanted into the right ocular vitreum, while the left control ocular vitreum received sham operation but nothing was implanted. SWI sequential test was made 2 hours postoperatively. Then the rabbits were killed and the specimens of the vitreous bodies of the rabbits were obtained. Hematoxylin and eosin staining and histological examinations were performed. RESULTS: The autologous eyelashes in 8 ocular vitreums of rabbits showed linear low signal intensity on the magnitude images and susceptibility weighted images, but linear high signal intensity on the phase images. Among the 12 experimental rabbits, 5 eyelashes in the right vitreum were completely shown in 3 rabbits, partly shown in 5 rabbits (2 eyelashes shown in 3 rabbits and 3 eyelashes shown in 2 rabbits), and not shown in 4 rabbits. CONCLUSIONS: SWI of the foreign body of intravitreous autologous eyelashes in rabbits has its own characteristics. The combined application of SWI sequential magnitude images, susceptibility weighted images and phase images is helpful to the detection and diagnosis of intravitreous autologous eyelashes in rabbits.

In vivo tracing of superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells transplanted for traumatic brain injury by susceptibility weighted imaging in a rat model.

OBJECTIVE: To label rat bone marrow mesenchymal stem cells (BMSCs) with superparamagnetic iron oxide (SPIO) in vitro, and to monitor the survival and location of these labeled BMSCs in a rat model of traumatic brain injury (TBI) by susceptibility weighted imaging (SWI) sequence. METHODS: BMSCs were cultured in vitro and then labeled with SPIO. Totally 24 male Sprague Dawley (SD) rats weighing 200-250 g were randomly divided into 4 groups: Groups A-D (n equal to 6 for each group). Moderate TBI models of all the rats were developed in the left hemisphere following Feeney's method. Group A was the experimental group and stereotaxic transplantation of BMSCs labeled with SPIO into the region nearby the contusion was conducted in this group 24 hours after TBI modeling. The other three groups were control groups with transplantation of SPIO, unlabeled BMSCs and injection of nutrient solution respectively conducted in Groups B, C and D at the same time. Monitoring of these SPIO-labeled BMSCs by SWI was performed one day, one week and three weeks after implantation. RESULTS: Numerous BMSCs were successfully labeled with SPIO. They were positive for Prussian blue staining and intracytoplasm positive blue stained particles were found under a microscope (200). Scattered little iron particles were observed in the vesicles by electron microscopy (5000). MRI of the transplantation sites of the left hemisphere demonstrated a low signal intensity on magnitude images, phase images and SWI images for all the test rats in Group A, and the lesion in the left parietal cortex demonstrated a semicircular low intensity on SWI images, which clearly showed the distribution and migration of BMSCs in the first and third weeks. For Group B, a low signal intensity by MRI was only observed on the first day but undetected during the following examination. No signals were observed in Groups C and D at any time points. CONCLUSION: SWI sequence in vivo can consecutively and noninvasively trace and demonstrate the status and distribution of BMSCs labeled with SPIO in the brain of TBI model rats.

[Formation and suppression technique for motion and flow artifacts on magnetic resonance imaging].

During magnetic resonance imaging, motion and flow artifacts often severely decrease image quality. The mechanisms of these kinds of artifacts are very complex. This article reviewed the pre-handling techniques of motion and flow artifacts, such as changing magnetic resonance imaging parameters, using spatial pre-saturation pulse sequences, gating and triggering techniques of suppressing respiratory motion artifact, as well as the progress of magnetic resonance pulse sequences in suppressing motion and flow artifacts, The paper briefly introduced post-processing techniques of suppressing motion and flow artifacts.

Melatonin reduces blood pressure in rats with stress-induced hypertension via GABAA receptors.

1. Several groups have reported that melatonin produces a significant decrease in blood pressure in mammals and that pinealectomy in rats causes hypertension. The purpose of the present study was to investigate the effects of melatonin and bicuculline methiodide on the blood pressure of rats, both in the developing and fully developed stage of stress-induced hypertension (SIH). 2. Rats with SIH were generated by mild electric foot shocks for 15 days, after which tail arterial systolic pressure and plasma angiotensin (Ang) II levels were measured. The effects of melatonin injections (i.p. or i.c.v.) on mean arterial pressure (MAP) in rats with SIH were also determined. 3. Pretreatment with 1 mg/kg, i.p., melatonin significantly diminished the elevated tail arterial systolic pressure and plasma AngII levels caused by 15 days stress. The suppressive effects of melatonin were blocked by i.p. injection of 1 mg/kg bicuculline methiodide, an antagonist of the GABA(A) receptor. 4. Intraperitoneal (0.2, 0.5 and 1 mg/kg) or i.c.v. (0.15 and 1.5 microg/3 microL) injection of melatonin produced a dose-dependent lowering of MAP in rats with SIH. The antihypertensive response induced by melatonin was blocked by injection of both 1 mg/kg, i.p., and 1.5 x 10(6) microg/3 microL, i.c.v., bicuculline methiodide. 5. In conclusion, melatonin not only prevents increases in blood pressure during the developing stage of SIH, but can also reduce the blood pressure of rats that have already developed SIH. The antihypertensive effect of melatonin may be mediated by GABA(A) receptors through inhibition of plasma AngII levels.