Long non-coding RNA CCAT1 promotes glioma cell proliferation via inhibiting microRNA-410.

BACKGROUND AND AIM: Long non-coding RNAs have been confirmed to play a critical role in various cancers. In the present study, the effect of long non-coding RNA (lncRNA) CCAT1 on glioma cell proliferation and its potential mechanism were investigated. METHODS AND RESULTS: Real-time PCR results showed that lncRNA-CCAT1 expression was significantly upregulated in glioma cancer tissues and cell lines compared with controls. After inhibiting CCAT1 expression in glioma cell line U251 with siRNA-CCAT1 (si-CCAT1), the cell viability and cell colony formation were decreased, the cell cycle was arrested in G1 phase, and the cell apoptosis was increased. As reported in bioinformatics software starbase2.0, a total of 22 microRNAs were potentially targeted by CCAT1. It was confirmed that miR-410 was altered most by si-CCAT1. After up-regulating CCAT1 expression in U251 cells, miR-410 level was decreased. Luciferase reporter assay confirmed that CCAT1 targeted miR-410. Correlation analysis showed that CCAT1 expression was negatively related to miR-410 expression in glioma cancer tissues. In addition, down-regulation of miR-410 reversed the inhibitory effect of si-CCAT1 on glioma proliferation. CONCLUSION: These data demonstrated that lncRNA-CCAT1 promoted glioma cell proliferation via inhibiting miR-410, providing a new insight about the pathogenesis of glioma proliferation.

TGF-ß induced miR-132 enhances the activation of TGF-ß signaling through inhibiting SMAD7 expression in glioma cells.

Transforming growth factors ß (TGF-ß) pathway has been proven to play important roles in oncogenesis and angiogenesis of gliomas. MiR-132 might be related to TGF-ß signaling pathway and high miR-132 expression was reported to be a biomarker of poor prognosis in patients diagnosed with glioma. However, the expression regulation way involved in TGF-ß pathway and clinical significance of miR-132 have not been investigated in glioma cells. Here we reported that the mRNA level of miR-132 and TGF-ß concentration were both increased in patients with brain glioma. Correlation analysis revealed that TGF-ß concentration was positively correlated with mRNA level of miR-132. In addition, the mRNA level of miR-132 was up-regulated by TGF-ß in a concentration-dependent and time-dependent manner. Furthermore, we found that miR-132 was involved in modulation of the TGF-ß signaling pathway and down-regulation of SMAD7 expression by directly targeting the SMAD7 3'-UTR. MiR-132 was negatively correlated with SMAD7 in patients with brain glioma. Taken together, our results suggest that miR-132 could be stimulated by TGF-ß and might enhance the activation of TGF-ß signaling through inhibiting SMAD7 expression in glioma cells. These findings contribute to a better understanding of the mechanism of the activation of TGF-ß signaling by miR-132.

CT-guided percutaneous microwave ablation of liver metastases from nasopharyngeal carcinoma.

PURPOSE: To access efficacy of percutaneous microwave ablation (MWA) of liver metastases from nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: From March 2007 to June 2012, 18 consecutive patients with NPC and liver metastases (15 men and three women; average age, 45.7 y; age range, 31-61 y) received computed tomography (CT)-guided percutaneous MWA treatment. A total of 27 ablations were performed involving 24 liver metastatic lesions in 18 patients with NPC. Average patient follow-up after ablation was 22.4 months (range, 2-52 mo). The average number of liver metastases per patient was 1.3 (range, 1-4 lesions), with lesion diameters ranging from 1.9 cm to 4.2 cm. Evaluation was then performed to assess percentage of complete necrosis, local tumor progression, and safety. RESULTS: Technical success was achieved in all 27 MWA procedures performed. During follow-up, new metastatic lesions developed in four of 18 patients. Of these, two were liver metastases, and were successfully treated with repeat WMA. Only two major complications were observed: pneumothorax in a patient with an ablation pathway involving the thorax and postprocedural pain in two other patients. A median overall survival time of 41.4 months was observed (range, 2-50 mo); three of 18 patients died during follow-up. CONCLUSIONS: CT-guided MWA is safe and offers an effective treatment alternative for local tumor control in selected patients with liver metastases from NPC.

Brachial plexus avulsion induced changes in gut microbiota promotes pain related anxiety-like behavior in mice.

Introduction: Brachial plexus avulsion (BPA) injury develops frequent and intense neuropathic pain, involving in both peripheral and central nervous systems. The incidence of anxiety or depression caused by BPA-induced neuropathic pain is high, but the underlying mechanism remains unclear. Methods: We established a BPA mice model and assessed its negative emotions through behavioral tests. To further explore the role of the microbiota-gut-brain axis in the unique emotional behavior after BPA, we performed intestinal fecal 16s and metabolomics assays. Psychobiotics (PB) supplementation was administered to BPA mice to check the probiotics effects on BPA-induced anxiety behaviors. Results: Pain related anxiety-like behavior was observed at the early stage after BPA (7 days), while no depression-like behavior was detected. Intriguingly, gut microbiota diversity was increased in BPA mice, and the most abundant probiotics, Lactobacillus, showed obvious changes. Lactobacillus_reuteri was significantly decreased in BPA mice. Metabolomics analysis showed that Lactobacillus_reuteri-related bile acid pathway and some neurotransmitter amino acids were significantly altered. Further PB (dominated by Lactobacillus_reuteri) supplementation could significantly relieve BPA-induced anxiety-like behaviors in mice. Conclusion: Our study suggests that pathological neuralgia after BPA could alter intestinal microbiota diversity, especially Lactobacillus, and the changes in neurotransmitter amino acid metabolites may be the key reason for the onset of anxiety-like behaviors in BPA mice.

Peripheral BDNF Regulates Somatosensory-Sympathetic Coupling in Brachial Plexus Avulsion-Induced Neuropathic Pain.

Brachial plexus avulsion (BPA) is a combined injury involving the central and peripheral nervous systems. Patients with BPA often experience severe neuropathic pain (NP) in the affected limb. NP is insensitive to the existing treatments, which makes it a challenge to researchers and clinicians. Accumulated evidence shows that a BPA-induced pain state is often accompanied by sympathetic nervous dysfunction, which suggests that the excitation state of the sympathetic nervous system is correlated with the existence of NP. However, the mechanism of how somatosensory neural crosstalk with the sympathetic nerve at the peripheral level remains unclear. In this study, through using a novel BPA C7 root avulsion mouse model, we found that the expression of BDNF and its receptor TrκB in the DRGs of the BPA mice increased, and the markers of sympathetic nervous system activity including α1 and α2 adrenergic receptors (α1-AR and α2-AR) also increased after BPA. The phenomenon of superexcitation of the sympathetic nervous system, including hypothermia and edema of the affected extremity, was also observed in BPA mice by using CatWalk gait analysis, an infrared thermometer, and an edema evaluation. Genetic knockdown of BDNF in DRGs not only reversed the mechanical allodynia but also alleviated the hypothermia and edema of the affected extremity in BPA mice. Further, intraperitoneal injection of adrenergic receptor inhibitors decreased neuronal excitability in patch clamp recording and reversed the mechanical allodynia of BPA mice. In another branch experiment, we also found the elevated expression of BDNF, TrκB, TH, α1-AR, and α2-AR in DRG tissues from BPA patients compared with normal human DRGs through western blot and immunohistochemistry. Our results revealed that peripheral BDNF is a key molecule in the regulation of somatosensory-sympathetic coupling in BPA-induced NP. This study also opens a novel analgesic target (BDNF) in the treatment of this pain with fewer complications, which has great potential for clinical transformation.

[Net power and energy of cooled antenna microwave ablation:ex vivo versus in vivo results in porcine liver].

OBJECTIVE: To explore the net power and net energy of a cooled antenna radiator in ex vivo and in vivo porcine livers. METHODS: All animal experiments complied with the guidelines of our animal use committee. Microwave ablation (MWA) was performed in ex vivo and in vivo porcine livers with a cooled-shaft antenna in different microwave ablation parameter groups (50, 80 and 110 W for 10 min). The energy losses from the microwave antenna or cables were calculated. And the net power, net energy and the relationship between net power and power readout were determined. RESULTS: When the power displayed by the machine indicated 50 W, 80 W and 110 W, the net power during MWA was 31.3 ± 0.6, 47.3 ± 0.8 and 62.1 ± 0.9 W ex vivo and 31.8 ± 0.8, 47.4 ± 0.3 and 61.7 ± 1.5 W in vivo. For the same power readout, the ex vivo or in vivo effective power was the same (P = 0.841, P = 0.133, P = 0.551). For both ex vivo and in vivo experiments, the ratio of microwave antenna energy loss to microwave antenna input energy was relatively constant (P = 0.613, 0.326). For the same treatment time and net power, the difference was significant between ex vivo and in vivo ablation volumes (P = 0.001, 0.006, 0.001). CONCLUSION: Using net power as a reference during MWA is more accurate compared to the traditional power readout. And net energy offers a more realistic reflection of MWA energy in tissues.

[Comparison between biliary cirrhosis and normal porcine liver treated with microwave ablation using a cooled-tip electrode].

OBJECTIVE: To elucidate the difference in both in vivo and ex vivo microwave ablation in a biliary cirrhotic porcine liver model using a cooled-tip antenna. METHODS: Two months after biliary ductal ligation, liver biopsy was performed to confirm the establishment of biliary cirrhosis in 4 Tibetan miniature pigs. Microwave ablation with cooled-tip antenna was conducted under laparotomy using 70 W for five minutes in the experimental group (4 pigs). The control group (2 pigs) also received microwave ablation using the same settings but no surgery. Both in-vivo and ex-vivo ablations were performed in the two groups. Morphological and pathological characteristics of the ablation areas were compared. Paired comparison among the groups were conducted using t-test. RESULTS: In the cirrhotic liver group, after ablation at 70 W for five minutes, the short and long axes and volume of in vivo ablation areas were (1.90 ± 0.10) cm, (2.95 ± 0.12) cm, and (6.0 ± 0.8) cm(3) compared to (2.08 ± 0.08) cm, (3.08 ± 0.75) cm, and (7.0 ± 0.5) cm(3) of ex vivo ablation. In the normal liver group the dates were (2.04 ± 0.05) cm, (3.14 ± 0.11) cm and (6.8 ± 0.5) cm(3); (2.30 ± 0.18) cm, (3.60 ± 0.08) cm and (10.0 ± 1.7) cm(3), respectively. In vivo ablation area was smaller than ex vivo ablation area in terms of short and long axes and volume (P = 0.028 0.026, 0.008, respectively). With the same ablation settings, both in vivo and ex vivo ablation areas in normal pig liver were larger than their counterparts in cirrhotic liver in terms of the short and long axes and volume (P = 0.019, P = 0.000; P = 0.024, P = 0.036, respectively), but the differences in the short axes of in vivo and ex vivo ablation areas failed to reach significance. CONCLUSION: Both in vivo and ex vivo ablation areas in biliary cirrhotic pig liver were smaller than their counterparts in normal pig liver suggesting that, the ablation time or power should be relatively prolonged to enlarge the ablation zone within cirrhotic liver in order to prevent incomplete ablation with viable residual tumor.

3D Printing Technology in Planning Thumb Reconstructions with Second Toe Transplant.

OBJECTIVE: To report preoperative planning using 3D printing to plan thumb reconstructions with second toe transplant. METHODS: Between December 2013 and October 2015, the thumbs of five patients with grade 3 thumb defects were reconstructed using a wrap-around flap and second toe transplant aided by 3D printing technology. CT scans of hands and feet were analyzed using Boholo surgical simulator software (www.boholo.com). This allowed for the creation of a mirror image of the healthy thumb using the uninjured thumb. Using 3D images of the reconstructed thumb, a model of the big toe and the second toe was created to understand the dimensions of the donor site. This model was also used to repair the donor site defect by designing appropriate iliac bone and superficial circumflex iliac artery flaps. The polylactic acid model of the donor toes and reconstructed thumb was produced using 3D printing. Surgically, the wrap-around flap of the first dorsal metatarsal artery and vein combined with the joint and bone of the second toe was based upon the model donor site. Sensation was reconstructed by anastomosing the dorsal nerve of the foot and the plantar digital nerve of the great toe. Patients commenced exercises 2 weeks after surgery. RESULTS: All reconstructed thumbs survived, although partial flap necrosis occurred in one case. This was managed with regular dressing changes. Patients were followed up for 3-15 months. The lengths of the reconstructed thumbs are 34-49 mm. The widths of the thumb nail beds are 16-19 mm, and the thickness of the digital pulp is 16-20 mm. The thumb opposition function was 0-1.5 cm; the extension angle was 5°-20° (mean, 16°), and the angle of flexion was 38°-55° (mean, 47°). Two-point discrimination was 9-11 mm (mean, 9.6 mm). The reconstructed thumbs had good appearance, function and sensation. Based on the criteria set forth by the Standard on Approval of Reconstructed Thumb and Finger Functional Assessment of the Chinese Medical Association, the results were considered excellent for four cases and good for one case. The success rate was 100%. CONCLUSIONS: When planning a wrap-around flap and second toe transplant to reconstruct a thumb, both the donor and recipient sites can be modeled using 3D printing. This can shorten the operative time by supplying digital and accurate schematics for the operation. It can also optimize the function and appearance of the reconstructed thumb while minimizing damage to the donor site.

Beta-asarone protects against MPTP-induced Parkinson's disease via regulating long non-coding RNA MALAT1 and inhibiting α-synuclein protein expression.

OBJECTIVE: Numerous long non-coding RNAs (lncRNA) have been identified in neurodegenerative disorders including Parkinson's disease (PD). Emerging evidence demonstrates that ß-asarone functions as neuroprotective effects in both in vitro and in vivo models. However, the role of ß-asarone and its potential mechanism in PD remain not completely clear. METHODS: MPTP-induced PD mouse model and SH-SY5Y cells subjected to MPP+ as its in vitro model were used to evaluate the effects of ß-asarone on PD. LncRNA MALAT1 and α-synuclein expression were determined by real-time PCR and western blot methods. RESULTS: ß-Asarone significantly increased the TH+ cells number and decreased the expression levels of MALAT1 and α-synuclein in midbrain tissue of PD mice. RNA pull-down and immunoprecipitation assays confirmed that MALAT1 associated with α-synuclein, leading to the increased stability of α-synuclein and its expression in SH-SY5Y cells. ß-asarone elevated the viability of cells exposed to MPP+. Either overexpressed MALAT1 or α-synuclein could canceled the protective effect of ß-asarone on cell viability. In PD mice, pcDNA-MALAT1 also decreased the TH+ cells number and increased the α-synuclein expression in PD mice with treatment of ß-asarone. CONCLUSION: ß-Asarone functions as a neuroprotective effect in both in vivo and in vitro models of PD via regulating MALAT1 and α-synuclein expression.

MicroRNA-214 participates in the neuroprotective effect of Resveratrol via inhibiting α-synuclein expression in MPTP-induced Parkinson's disease mouse.

BACKGROUNDS AND AIMS: MicroRNAs (miRNAs) have been reported to be involved in degenerative disorders including Parkinson's disease (PD). α-synuclein expression is strong associated with the pathogenesis of PD. In the present study, we investigated whether the regulation of α-synuclein expression by miR-214 is the potential mechanism underlying the neuroprotective effect of Resveratrol. METHODS: The PD mouse model was established with the injection of MPTP (1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and the human neuroblastoma cell line, SH-SY5Y, was administrated with MPP+. RESULTS: The midbrain of PD mice and MPP+ treated SH-SY5Y cells had the lower expression levels of miR-214 and higher mRNA and protein expression of α-synuclein, which were reversed by Resveratrol administration. MiR-214 mimic down-regulated expression of α-synuclein and its 3'-UTR activity, while the levels were up-regulated by miR-214 inhibitor. In addition, the cell viability, elevated by Resveratrol, was also decreased by miR-214 inhibitor or overexpressed α-synuclein. In vivo, miR-214 inhibitor down-regulated TH+ cells of ipsilateral and up-regulated α-synuclein expression compared with the group treated with Resveratrol. CONCLUSION: The loss of miR-214 in PD resulted in the increase of α-synuclein expression, which was the potential mechanism underlying the neuroprotective effects of Resveratrol.

[Free perforating flap of peroneal artery for repairing the forefoot skin defects].

OBJECTIVE: To explore the clinical techniques and effects of repairing skin defects of the forefoot by free perforating flap nourished by peroneal artery. METHODS: From June 2007 to June 2011, 11 patients with skin and soft tissue defects of the forefoot were repaired by free peroneal artery perforating flap in emergent or subemergent. There were 10 males and 1 female with an average age of 28.6 years old ranging from 23 to 46 years old. Among them, 4 cases injured for traffic accidents, 3 for crush and 4 for machine strangulation. In all cases, the defect area of forefoot tissue varied from 2.0 cm x 4.0 cm to 4.0 cm x 8.5 cm,and the adopted area varied from 2.5 cm x 4.5 cm to 4.0 cm x 9.0 cm. The operation time was from 6 to 96 h (averaged 31.8 h). The blood vessels were anastomosed end-to-end. RESULTS: All of the transferred free flaps survived uneventfully. Nine of them were successfully followed up from 6 to 24 months. The appearance, elasticity and functions of flaps were satisfied accompanied with slight damage of donor site although seemed bloated. The smaller donor site could be intimately seamed if necessary. CONCLUSION: The vessels anatomy of knee with antegrade extended peroneal artery was relative constant with a moderate thickness and simple operation, is useful to repair small or middle areas of skin defects in forefoot.