PI3K-Akt-mTOR inhibition by GNE-477 inhibits renal cell carcinoma cell growth in vitro and in vivo.

Sustained activation of PI3K-Akt-mTOR cascade is important for renal cell carcinoma (RCC) cell progression. GNE-477 is a novel and efficacious PI3K-mTOR dual inhibitor. The current study tested its anti-RCC cell activity. In the primary cultured human RCC cells, GNE-477 potently inhibited cell growth, viability and proliferation, as well as cell cycle progression, migration and invasion. Furthermore, it induced robust apoptosis activation in primary RCC cells, but being non-cytotoxic to HK-2 epithelial cells and primary human renal epithelial cells. In the primary RCC cells GNE-477 inactivated PI3K-Akt-mTOR cascade by blocking phosphorylation of p85, Akt1, p70S6K1 and S6. Restoring Akt-mTOR activation by a constitutively-active Akt1 reversed GNE-477-induced anti-RCC cell activity. In nude mice intraperitoneal injection of GNE-477 potently suppressed RCC xenograft tumor growth. Collectively, targeting PI3K-Akt-mTOR cascade by GNE-477 inhibits RCC cell growth in vitro and in vivo.

[Effect of changes of inflammatory factors on prognosis of tibia plateau Schatzker â ¢ fractures treated under arthroscopic or open reduction and internal fixation].

OBJECTIVE: To explore changes of inflammatory factors on prognosis of tibia plateau Schatzker Ⅲ fractures treated under arthroscopic or open reduction and internal fixation. METHODS: From November 2013 to November 2016, clinical data of 30 patients with tibia plateau Schatzker Ⅲ fractures were retrospectively analyzed, and divided into minimally invasive group and control group according to different surgical methods 15 patients in each group. Minimally invasive group were treated by arthroscopic internal fixation, including 8 males and 7 females, aged from 20 to 50 years old with an average of (35.0± 14.6) years old, the time from injury to operation ranged from 7 to 15 days with an average of (11.0±4.1) days. Control group were treated by open reduction and internal fixation, including 7 males and 8 females, aged from 18 to 48 years old with an average of (33.0±13.6) years old, the time from injury to operation ranged from 6 to 14 days with an average of (10.0±3.4) days.Operation time, length of incision, blood loss, postoperative loading time and fracture healing time, complications were compared between two groups. Level of IL-1ß, IL-6, TNF-α were detected at 3 days, 6 months and 12 months after operation, Lysholm knee function score at 6 and 12 months were compared between two groups. RESULTS: Allpatients were followed up, but there was no significant difference in following up between two groups. Operation time, length of incision, blood loss, postoperative loading time, fracture healing time and cases of complications in minimally invasive group were (80.3±9.7) min ,(4.2± 1.0) cm ,(102.2±26.4) ml ,(30.0±10.0) d ,(70.0±5.0) d and 0 case respectively; while in control group were (90.3±9.1) min, (10.5±1.1) cm ,(221.1±46.8) ml ,(50.0±15.0) d ,(90.0±6.0) d and 2 cases respectively; there were significant difference between two groups. Lysholm score in minimally invasive group 89.2±5.1 was higher than that of control group 80.1±3.1; and score of swelling, squat and pain in minimally invasive group was higher than that of control group at 6 months after opertaion. While there were no significant difference in each items and total score of Lysholm score between two groups at 12 months after operation. Level of IL-1ß, IL-6, and TNF-α in minimally invasive group at 3 days and 6 months were [(52.1±20.1) pg/L, (0.9±0.1) pg/L ],[(56.1±20.1) pg/L ,(1.1±1.3) pg/L ] and [(28.3±2.5) pg/L ,(8.4±1.5) pg/L ] respectively; while in control group were [(64.8±9.1) pg/L ,(8.1±2.1) pg/L ],[(65.8±12.3) pg/L ,(9.1±5.3) pg/L ] and [(38.5±2.3) pg/L ,(26.5± 1.4) pg/L ] respectively; there were statistically difference in level IL-1ß, IL-6 and TNF-α between two groups at 3 days and 6 months after operation; while there was no difference at 12 months after operation (P>0.05) . Inflammatory cytokines level at 3 days after operation IL-1ß [OR=1.279, 95%CI (1.047, 1.512), P<0.05 ], IL-6 [OR=1.687, 95%CI (1.478, 1.888), P<0.05 ], TFN-α [ OR=2.096, 95%CI (1.863, 2.316), P<0.05 ] was an independent risk factor for Lysholm knee function score at 6 months after operation. CONCLUSION: Arthroscopic surgery and open surgery also could obtain good clinical effects in treating tibia plateau Schatzker Ⅲ fractures. Arthroscopic internal fixation could shorten operation time, lessen the mount of blooding with minimally invasive, lower occurrence of postoperative complications, faster recovery of knee function.

The therapeutic value of the SphK1-targeting microRNA-3677 in human osteosarcoma cells.

Sphingosine kinase 1 (SphK1) is a potential therapeutic target for human osteosarcoma (OS). SphK1-targeting microRNAs (miRNAs) could have important therapeutic value for OS. We discovered that micorRNA-3677 (miR-3677) is a SphK1-targeting miRNA, inhibiting OS cell progression. The results of RNA-Pull down assay confirmed direct binding between biotinylated-miR-3677 and SphK1 mRNA in primary human OS cells. In established and primary human OS cells forced overexpression of miR-3677, by a lentiviral construct, decreased SphK1 3'-UTR (untranslated region) activity and downregulated SphK1 expression. Both were however enhanced with miR-3677 inhibition in OS cells. Function studies demonstrated that OS cell growth, proliferation and migration were inhibited with miR-3677 overexpression, but augmented with miR-3677 inhibition. MiR-3677 overexpression-induced anti-OS cell activity was reversed with re-expression of the 3'-UTR-depleted SphK1. Additionally, in SphK1 knockout OS cells (by CRISPR/Cas9 strategy), altering miR-3677 expression failed to further alter cell functions. Finally, we show that miR-3677 expression was significantly downregulated in primary human OS tissues, correlating with SphK1 mRNA upregulation. We conclude that targeting SphK1 by miR-3677 inhibits human OS cell progression.

[Ultrasound-guided minimal traverse-cross technique repair for acute closed Achilles tendon ruptures].

OBJECTIVE: To explore clinical effects of ultrasound-guided minimal traverse-cross technique repair for acute closed Achilles tendon ruptures. METHODS: From January 2015 to March 2017, 20 patients with acute closed Achilles tendon rupture were treated by minimal traverse-cross technique repair with ultrasound guided. Among them, including 13 males and 7 females, aged from 28 to 49 years old with an average of(31.3 ±4.5) years old. All patients were single side injury. Fifteen patients on the left side and 5 patients were on the right side. The time from injury to operation ranged from 1 to 5 days with an average of (2.5±0.7) days. Operative time, postoperative complications were observed, and AOFAS score before and after operation at 12 months were compared. RESULTS: All patients were followed up for 12 to 27 months with an average of(15.2±4.9) months. Operative time ranged from 33 to 65 min with an average of(43.7±5.6) min. Incision of one patient were continued oozing and improved after changing dressings, other patients were healed at stage I. No sural nerve irritation symptoms and palindromic rapture of heel tendon occurred. AOFAS score was improved from 65.2±7.4 before operation to 97.7±4.7 after operation at 12 months (t=22.5, P<0.01); 18 patients got excellent results and 2 good. CONCLUSIONS: Ultrasound-guided minimal traverse-cross technique repair for acute closed Achilles tendon ruptures, which promise minimal incision, protect sural nerve, ensure quality of tendon anastomosis and fixation, and is a ideal method for repairing acute closed Achilles tendon ruptures.

[Analysis of hidden blood loss after percutaneous kyphoplasty for osteoporotic vertebral compression fracture and its influencing factors].

OBJECTIVE: To investigate the hidden blood loss and related risk factors of osteoporotic vertebral compression fractures after percutaneous kyphoplasty. METHODS: The clinical data of 153 patients with osteoporotic vertebral compression fractures who underwent percutaneous kyphoplasty from March 2015 to December 2017 were retrospectively analyzed, including 55 males and 98 females, aged 68 to 87(78.6±11.4) years old. Erythrocyte specific volume was collected before and after operation to calculate the hidden blood loss. The influence of sex, age, body mass index, bone mineral density, diabetes mellitus and hypertension, operation mode (unilateral or bilateral), operation time, operative segment and number, loss height of vertebral body and recovery height ratio on hidden blood loss was analyzed by multiple linear regression model. RESULTS: Postoperative hidden blood loss was (287.7±68.5) ml. Multivariate linear regression analysis showed that the history of diabetes mellitus (ß=2.405, P=0.012), the mode of operation(ß=3.042, P<0.001), the time of operation (ß=2.043, P=0.038), the operative segment (ß=1.993, P=0.043), the number (ß=0.374, P<0.001), the loss of vertebral height (ß=2.785, P=0.003) and the recovery ratio(ß=7.301, P<0.001) were correlated with occult hemorrhage. CONCLUSIONS: There is a certain degree of occult hemorrhage in kyphoplasty for osteoporotic vertebral compression fractures. The risk factors of hidden hemorrhage are diabetes history, operation method, operation time, operative segment and number, loss of vertebral height and recovery ratio.

microRNA-29a inhibition induces Gab1 upregulation to protect OB-6 human osteoblasts from hydrogen peroxide.

The present study determines the role of the Gab1 in hydrogen peroxide (H2O2)-induced death of human osteoblasts. We show that Gab1 is required for H2O2-induced Akt activation to promote osteoblast survival. In OB-6 human osteoblasts, Gab1 silencing (by targeted-shRNA) or complete knockout (by CRISPR-Cas9 KO plasmid) largely attenuated Akt activation by H2O2. Gab1-depleted OB-6 cells were more vulnerable to H2O2. Conversely, forced over-expression of Gab1 by an adenovirus vector increased Akt activation to protect OB-6 cells from H2O2. Significantly, the anti-sense of microRNA-29a ("antagomiR-29a") induced Gab1 expression to facilitate H2O2-induced Akt activation, which protected OB-6 cells from apoptosis. AntagomiR-29a was however ineffective in Gab1-deficient and Akt-inhibited OB-6 cells. Forced over-expression of miR-29a induced Gab1 downregulation to inhibit H2O2-induced Akt activation, causing enhanced OB-6 cell death. miR-29a-induced actions were abolished by an adenovirus constitutively-active Akt1 (Ad-caAkt1) in OB-6 cells. Together, microRNA-29a inhibition induces Gab1 upregulation and Akt activation to protect OB-6 osteoblasts from H2O2.

SC79 rescues osteoblasts from dexamethasone though activating Akt-Nrf2 signaling.

Dexamethasone (Dex) causes osteoblast cell injuries. In the present research, we tested the potential effect of SC79, a novel and specific Akt activator, against Dex in osteoblasts. In primary murine osteoblasts and osteoblastic MC3T3-E1 cells, pretreatment with SC79 significantly attenuated Dex-induced cell death. Further, Dex-induced mitochondrial permeability transition pore (mPTP) opening, cytochrome C release and apoptosis activation were dramatically alleviated with SC79 pretreatment in above cells. At the molecular level, SC79 activated Akt, which was indispensable for subsequent osteoblast protection against Dex. Akt inhibitors (LY294002, perifosine and MK-2206) blocked SC79-induced Akt activation and abolished its anti-Dex actions in osteoblasts. Further, SC79 activated Akt downstream Nrf2 (NF-E2-related factor 2) signaling and attenuated Dex-induced oxidative stress in osteoblasts. Nrf2 shRNA knockdown or S40T mutation almost reversed SC79-mediated anti-oxidant and cytoprotective activities in osteoblasts. Together, these results suggest that SC79 activates Akt-Nrf2 signaling to protect osteoblasts from Dex.

miR-135b expression downregulates Ppm1e to activate AMPK signaling and protect osteoblastic cells from dexamethasone.

Activation of AMP-activated protein kinase (AMPK) could potently protect osteoblasts/osteoblastic cells from dexamethasone (Dex). We aim to induce AMPK activation via microRNA ("miRNA") downregulation of its phosphatase Ppm1e. We discovered that microRNA-135b ("miR-135b") targets the 3' untranslated regions (UTRs) of Ppm1e. In human osteoblasticOB-6 cells and hFOB1.19 cells, forced-expression of miR-135b downregulated Ppm1e and activated AMPK signaling. miR-135b also protected osteoblastic cells from Dex. shRNA-induced knockdown of Ppm1e similarly activated AMPK and inhibited Dex-induced damages. Intriguingly, in the Ppm1e-silenced osteoblastic cells, miR-135b expression failed to offer further cytoprotection against Dex. Notably, AMPK knockdown (via shRNA) or dominant negative mutation abolished miR-135b-induced AMPK activation and cytoprotection against Dex. Molecularly, miR-135b, via activating AMPK, increased nicotinamide adenine dinucleotide phosphate (NADPH) activity and inhibited Dex-induced oxidative stress. At last, we found that miR-135b level was increased in human necrotic femoral head tissues, which was correlated with Ppm1e downregulation and AMPK activation. There results suggest that miR-135b expression downregulates Ppm1e to activate AMPK signaling, which protects osteoblastic cells from Dex.

[Mini-open repair for the treatment of acute closed achilles tendon ruptures].

OBJECTIVE: To evaluate the clinical efficacy of mini-open repair for the treatment of acute closed achilles tendon ruptures. METHODS: From April 2012 and October 2013,14 patients (14 feet) with acute closed achilles tendon ruptures were treated in our department. They were 9 males and 5 females, with an average age of 30.5 years old (ranged, 25 to 49 years old). The interval between injury and operation ranged from 1 to 13 days (8 days on average). A longitudinal incision approximately 1.5 to 2.0 cm in length was made around the ruptured achilles tendon for mini-open repair after insertion of oval clamp. Postoperative rehabilitation was carried out. RESULTS: The wounds healed at the first stage except 2 cases with slow recovery. All the patients were followed up for 6 to 24 months, with an average of 11 months. According to the ankle-hindfoot scoring system of American Orthopaedic Foot & Ankle Society (AOFAS),the score was 92.71 ± 6.58 (82 to 100). CONCLUSION: The surgical treatment of acute achilles tendon rupture with mini-open repair has advantages of little invasion, a low rate of incision problems, quick function recovery, and simple operation, and it is suitable for primary hospital.

A reduction clamp for an aiming component in associated acetabular fractures.

BACKGROUND: The treatment of acetabular fractures is complex and requires specialized equipment. However, all currently available instruments have some disadvantages. A new reduction clamp that can firmly enable reduction and not hinder subsequent fixation procedures for some special fracture types is needed. MATERIALS AND METHODS: In this study, we introduce a new acetabular clamp and its preliminary clinical application in three T-shaped acetabular fractures. RESULTS: This new clamp can successfully pull the posterior column back to the anterior column and firmly maintain the reduction. This clamp's aiming plate can facilitate the insertion of long lag screws. The clamp is also easy to assemble and use. CONCLUSION: This reduction clamp is a useful instrument that can facilitate open reduction and internal fixation of acetabular fractures.

[Clinical application progress of hip arthroscopy].

The hip arthroscopy develops quickly in the past 20 years. Its use in the diagnosis and treatment of the hip injury get more notably. Many of pathologic conditions previously unrecognized through X-ray, CT or MRI have been diagnosed correctly under hip arthroscopy. The technology has become gradually a golden standard to confirm the hip discords. In the present, the examining path of the hip arthroscopy contains mainly the outside path, the anterior path, the outside-posterior path. The application range of the hip arthroscopy includes the management of labral tears, the femoroacetabular impingement, the ligamentum tear injuries, the chondral lesions, the synovical abnormalities, the intra-articular infection, the loose bodies in the joint etc. The hip arthroscopy is a comprehensive technology. Its superiority can be reflect well if the doctors are familiar with the indication, the contraindication and the operation procedure. And also the thorough physical examination and imaging examination should be made before operation, as well as reasonable and effective postoperative function training should be conducted.

[Treatment of atlantoaxial instability with C1-C2 posterior transarticular screw fixation].

OBJECTIVE: To evaluate the technique of C1-C2 transarticular screw fixation for atlantoaxial instability or dislocation. METHODS: Among 14 patients with atlantoaxial instability, 10 were male and 4 patients were female, with an average age of 38.6 years ranging from 17 to 62 years. All patients were treated by internal fixation with transarticular screws. RESULTS: There were 28 screws applied in 14 patients. All patients were followed up. The average follow-up period was 16 months (range, 9 to 35 months). The postoperative JOA score was ranging from 13.8 to 15.8 with the average score of (14.50 +/- 0.66) and the improved rate of (76.12 +/- 4.94)%. No spinal injury and vertical artery injury was found and osseous fusion was completed in all patients. CONCLUSION: The technique of C1-C2 transarticular screw fixation is one of the best treatments for atlantoaxial instability. Without the help of structural bone graft and aided internal fixation, morselized cancellous bone graft can acquire effective osseous fusion.