MicroRNA-141-3p promoted the progression of nasopharyngeal carcinoma through targeting DLC1.

OBJECTIVE: Previous studies have shown that the function of miR-141 has tissue specificity. However, the role of miR-141-3p has not been reported in nasopharyngeal carcinoma (NPC). Therefore, this study explored the function of miR-141-3p in NPC. PATIENTS AND METHODS: MiR-141-3p expression in NPC tissues was examined via quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. Cell Counting Kit-8 (CCK-8) and transwell assays were used to explore the function of miR-141-3p. The relationship between miR-141-3p and DLC1 was verified by Dual-Luciferase assay. Protein expression was observed by immunocytochemical assay and Western blot analysis. RESULTS: Upregulation of miR-141-3p associated with poor prognosis was detected in NPC patients. Moreover, overexpression of miR-141-3p promoted cell proliferation, migration, and invasion in NPC cells. It was also found that miR-141-3p promoted EMT and activated the mTOR signaling pathway in NPC. Furthermore, DLC1 was indicated as a direct target of miR-141-3p and miR-141-3p negatively correlated with DLC1 expression in NPC. In particular, upregulation of DLC1 could impair the promoted effect of miR-141-3p in NPC. CONCLUSIONS: MiR-141-3p promotes the progression of NPC by targeting DLC1 and activating the mTOR pathway.

Effects of oxaliplatin on inflammation and intestinal floras in rats with colorectal cancer.

OBJECTIVE: The aim of this study was to investigate the effects of oxaliplatin on intestinal floras, inflammation level, apoptosis-related gene expressions and oxidative stress in rats with colorectal cancer. MATERIALS AND METHODS: A total of 30 adult Sprague-Dawley (SD) rats were selected as research objects and were divided into control group, model group and oxaliplatin group. Rats in control group were raised normally, without any treatment. Rats in model group were subcutaneously injected with dimethylhydrazine (25 mg/kg) to establish the model of colorectal cancer. Meanwhile, rats in oxaliplatin group were injected with oxaliplatin (15 mg/kg) once every 2 weeks for 12 consecutive weeks. Peripheral blood, intestinal tumor tissues and feces were collected from rats. In addition, inflammatory indexes [tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), interleukin-4 (IL-4) and IL-1ß], oxidative stress indexes [catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and total antioxidant capacity (T-AOC)], expressions of apoptosis-related genes [apoptotic protease activating factor-1 (Apaf1), Caspase-9, Survivin and B-cell lymphoma-2 (Bcl-2)] and intestinal floras were detected. RESULTS: The abundance of microorganisms such as Sphaerobacterales, Adlercreutzia and Coriobacterium glomerans increased significantly in the intestines in control group (p<0.05). However, the abundance of Bifidobacterium, Rikenellaceae and Paraprevotella in the intestines was obviously higher in model group (p<0.05). Oxaliplatin group exhibited remarkably higher abundance of such microorganisms as Cyanobacteria, Alistipes and Metascardovia in rat intestines (p<0.05). The content of Alistipes was the highest in oxaliplatin group, followed by control group and model group, and the difference was statistically significant (p<0.05). The levels of serum TNF-α, CRP and IL-1ß were remarkably higher in model group than those in control group (p<0.05). Oxaliplatin group exhibited notably lower levels of serum TNF-α, CRP and IL-1ß (p<0.05) and higher IL-4 level than model group (p<0.05). The content of serum CAT, SOD, GSH and T-AOC was markedly elevated in model group compared with control group (p<0.05). However, it was significantly reduced in oxaliplatin group in comparison with model group (p<0.05). Compared with control group, model group had distinctly lower expressions of Apaf1, Caspase-9 and Survivin but an evidently higher expression level of Bcl-2 in tumor tissues (p<0.05). Moreover, the expressions of Apaf1, Caspase-9 and Survivin were clearly higher, while that of Bcl-2 was prominently lower in tumor tissues in oxaliplatin group than model group (p<0.05). CONCLUSIONS: Oxaliplatin exerts significant effects on the inflammation, oxidative stress, apoptosis-related genes and intestinal floras in rats with CRC.

Exogenous IL-19 mediates downregulation of TGF-ß through Erk and p38 pathway to inhibit epidural fibrosis.

OBJECTIVE: To evaluate the effect of interleukin-19 (IL-19) treatment on epidural fibrosis and its mechanism of action with transforming growth factor ß (TGF-ß). MATERIALS AND METHODS: Initially, IL-19 (10, 20, 50 and 100 ng/L) was used to pretreat rat fibroblasts. TGF-ß (10 µg/L) was then applied to activate fibroblasts. The protein expression levels of TGF-ß 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-ß 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. RESULTS: Concentration-dependent IL-19 significantly down-regulated TGF-ß receptor expression and inhibited phosphorylated Erk (p-Erk) and phosphorylated p38 (p-p38). In vivo, IL-19 reduced the expressions of TGF-ß 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. CONCLUSIONS: IL-19 inhibited TGF-ß 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.

Effect of miR-29b on rats with gestational diabetes mellitus by targeting PI3K/Akt signal.

OBJECTIVE: To investigate the role of micro-ribonucleic acid-29b (miR-29b) in rats with gestational diabetes mellitus (GDM) through the phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinase (Akt) signal and its mechanism by establishing rat models of GDM. MATERIALS AND METHODS: Rat models of GDM were constructed, and then the expression levels of miR-29b, total PI3K, phosphorylated PI3K (p-PI3K), total Akt and phosphorylated Akt (p-Akt) in the model group and control group were measured via Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting assays, and the association between miR-29b expression and total PI3K expression was analyzed. In addition, miR-29b mimics and inhibitors were used to further explore the regulatory pathway, and the influences of miR-29b mimics and inhibitors on PI3K and Akt phosphorylation in GDM rats, characteristic indicators of oxidative stress such as superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in liver tissues of GDM rats, and fasting blood glucose in GDM rats were studied. RESULTS: Compared with those in the control group, miR-29b expression was lowered in rat models of GDM, while PI3K/Akt signal expression was increased. In rats with GDM, miR-29b expression was prominently negatively correlated with total PI3K expression (r=-0.777, p=0.007, p<0.01). MiR-29b mimics could reduce PI3K and Akt phosphorylation, increase SOD and CAT expression levels and decrease MDA content (p<0.05). Moreover, miR-29b mimics significantly lowered the blood glucose level in rats with GDM (p<0.05). CONCLUSIONS: MiR-29b mimics can alleviate oxidative stress and reduce blood glucose by inhibiting the PI3K/Akt signal transduction.

Geometric Parameter Analysis of Ruptured and Unruptured Aneurysms in Patients with Symmetric Bilateral Intracranial Aneurysms: A Multicenter CT Angiography Study.

BACKGROUND AND PURPOSE: Previous studies of geometric and morphologic parameters of intracranial aneurysms have been conducted to determine rupture risk, which remains incompletely defined due to patient-specific risk factors, such as sex, hypertension, and age. To this end, we compared characteristics of ruptured and unruptured aneurysms in the same patients with symmetric bilateral intracranial aneurysms. MATERIALS AND METHODS: Between January 2008 and March 2014, 2361 patients with 2674 aneurysms were diagnosed by CT angiography or surgical findings at 4 medical centers. Geometric and morphologic parameters examined for symmetric bilateral intracranial aneurysms comprised aneurysm wall regularity, size, neck width, aspect ratio, size ratio, neck-to-parent artery ratio, and area ratio. Univariate and multivariate statistical analyses were performed to determine independent risk factors for rupture. RESULTS: Sixty-three patients (48 women, 15 men; mean age, 62.5 ± 9.8 years) with symmetric bilateral aneurysms were eligible for the study and were included. The most frequent aneurysm location was the posterior communicating artery. Univariate analysis disclosed that aneurysm size, aspect ratio, size ratio, area ratio, and irregular wall differed between patients with ruptured and unruptured aneurysms. Multivariate analysis indicated that aspect ratio of ≥1.6 (adjusted OR, 9.521; 95% CI, 2.182-41.535), area ratio of ≥1.5 (adjusted OR, 4.089; 95% CI, 1.247-13.406), and irregular shape (adjusted OR, 10.443; 95% CI 3.394-32.135) were significant predictive factors for aneurysm rupture after adjustment for aneurysm size. CONCLUSIONS: An aspect ratio of ≥1.6, area ratio of ≥1.5, and irregular wall are associated with aneurysm rupture independent of aneurysm size and patient characteristics. These characteristics alone can help in distinguishing ruptured bilateral intracranial aneurysms from unruptured ones.

High frequency of OTOF mutations in Chinese infants with congenital auditory neuropathy spectrum disorder.

Auditory neuropathy spectrum disorder (ANSD) is one of the most common diseases leading to hearing and speech communication barriers in infants and young children. The OTOF gene is the first gene identified for autosomal recessive non-syndromic ANSD, and patients with OTOF mutations have shown marked improvement of auditory functions from the cochlear implantation, but the true involvement of OTOF mutations in Chinese ANSD patients is still unknown which precludes the effective management of this disease. Here, we investigated the contribution of OTOF mutations to congenital ANSD patients in China. In all, 37 infants and young Children with ANSD were screened for all the exons of OTOF gene, of them 34 patients had no neonatal risk factors who were considered as congenital ANSD. The clinical manifestation and audiometric features were also investigated and compared in patients with and without OTOF mutations. In all, 14 of these subjects were shown to carry two or three mutant alleles of OTOF with the high frequency of 41.2% in congenital ANSD patients. In total, 15 novel pathogenic mutations and 10 reported mutations were identified. Our results confirmed that mutations in OTOF gene were a major cause of congenital ANSD in China. Identification of OTOF mutations can facilitate diagnosis, clinical intervention and counseling for congenital ANSD.

Mechanisms of cytotoxicity induced by the anesthetic isoflurane: the role of inositol 1,4,5-trisphosphate receptors.

Isoflurane can induce widespread cytotoxicity. We hypothesized that isoflurane induces apoptosis partly by causing excessive calcium release from the endoplasmic reticulum (ER) via direct activation of inositol 1,4,5-trisphosphate receptors (IP3R). Rat pheochromocytoma cells cultured for seven days with nerve growth factor were divided into four groups: control group (C), IP3R antagonist group (X), isoflurane group (I) and isoflurane + IP3R antagonist group (I+X). Groups I and I+X were treated with 1 MAC isoflurane for 12 h. Groups X and I+X were pretreated with IP3R antagonist. Annexin V/PI apoptosis and TUNEL assays were performed to evaluate cell apoptosis. TEM was used to observe changes in cell ultrastructure. Changes in calcium concentration ([Ca(2+)]i) in the cytoplasm were measured by flow cytometry. RT-PCR was performed to evaluate IP3R mRNA expression. TEM showed that isoflurane treatment altered cell ultrastructure. Compared to group C, cell apoptosis rate and [Ca(2+)]i increased in groups I and I+X (P < 0.05). Compared to group C, IP3R mRNA expression was lower in group X and higher in group I (P < 0.05). Compared to group X, cell apoptosis rate, [Ca(2+)]i and IP3R mRNA expression increased in groups I and I+X (P < 0.05). Compared to group I, cell apoptosis rate, [Ca(2+)]i and IP3R mRNA expression decreased in group I+X (P < 0.05). These results suggest that exposure to 1 MAC isoflurane for 12 h causes excessive calcium release partly by direct activation of IP3R on the ER membrane and triggers cell apoptosis.

Expression of RGC32 in human normal and preeclamptic placentas and its role in trophoblast cell invasion and migration.

INTRODUCTION: Preeclampsia (PE) is a pregnancy-specific complication and it is related to insufficient extravillous trophoblast invasion. To date, the pathophysiology of PE has not yet been fully elucidated. Response gene to complement 32 (RGC32) is a novel cellular protein, and it plays important roles in the regulation of cell differentiation, angiogenesis, migration, and invasion. This study aimed to determine the RGC32 expression and function in human placentas and to explore the underlying mechanisms. METHODS: RGC32 expression in term placentas collected after cesarean section from pregnant women with PE and normal pregnant women was determined by real-time reverse transcriptase polymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry. The effects of RGC32 expression on trophoblast invasion, migration, and the underlying mechanisms were studied in HTR8/SVneo cells. RESULTS: The messenger RNA (mRNA) and protein levels of RGC32 were significantly downregulated in preeclamptic placentas compared with normal controls (P < 0.05). RGC32 silencing significantly inhibited HTR8/SVneo cell migration and invasion (P < 0.001, respectively). These effects were associated with decreased activities and expression of matrix metalloproteinase (MMP)-2/9, and with the reduced phosphorylation level of Akt. DISCUSSION: RGC32 may play important roles in the pathophysiology of PE by directly affecting the invasion/migration of trophoblast.

Staged reimplantation for the treatment of fungal peri-prosthetic joint infection following primary total knee arthroplasty.

BACKGROUND: Fungal peri-prosthetic joint infections (PJI) are rare complication following total knee arthroplasty (TKA). There exists no established guidelines in the treatment of these infections and controversies are focused on the usefulness of antifungal-loaded cement spacers, the duration of systemic antifungal treatment and the ideal interval between implant removal and reimplantation. Therefore we ask if: (1) adding antifungal in cement space is a viable solution to manage fungal PJI; (2) there is no adverse effect adding antifungal medication in cement? HYPOTHESIS: We hypothesized that fungal PJI following TKA could be managed successfully by 2-stage reimplantation strategy using antifungal-loaded cement spacer. PATIENTS AND METHODS: Five cases of fungal PJI following total knee arthroplasty were treated in our institution between 2007 and 2013 using a 2-stage reimplantation strategy. The median elapsed time from primary arthroplasty to the diagnosis of infection was 7.4 months (range, 5-10 months). The infection was caused by Candida species in 4 cases and Pichia anomala in 1 case. Antibiotic- and antifungal-loaded articulating cement spacer was implanted during the interval between stages. Systemic antifungal agents were administered for at least 6 weeks after removal of prosthesis in all cases. The mean interval between removal and reimplantation was 6 months (range, 3-9 months). RESULTS: At a mean follow-up of 41.6 months (range, 24-65 months) after reimplantation, no patient had recurrent infection or revision due to any other reasons. The mean global IKS score improved from 58.4 (range, 37-96 points) preoperatively to 152.4 (range, 136-169 points) at final follow-up. The average range of motion of the knee for flexion improved from 63° (range, 10-110°) preoperatively to 98° (range, 80-120°) at final follow-up. CONCLUSIONS: Fungal PJI following TKA can be successfully treated by a staged reimplantation strategy. Antibiotic- and antifungal-loaded cement spacer implanted during interval period between stages may be an effective adjunct to therapy. Effective antifungal therapy is crucial to a successful result without adverse effect. LEVEL OF EVIDENCE: IV: retrospective or historical series.

C6-ceramide synergistically potentiates the anti-tumor effects of histone deacetylase inhibitors via AKT dephosphorylation and α-tubulin hyperacetylation both in vitro and in vivo.

Histone deacetylase inhibitors (HDACIs) have shown promising anti-tumor effects for a variety of malignancies, however, many tumors are reportedly resistant to them. In this study, we made a novel discovery that co-administration of HDACIs (Trichostatin A (TSA) and others) and exogenous cell-permeable short-chain ceramide (C6) results in striking increase in cancer cell death and apoptosis in multiple cancer cells. These events are associated with perturbations in diverse cell signaling pathways, including inactivation of Akt/mTOR and increase in α-tubulin acetylation (both in vivo and in vitro). TSA interacts in a highly synergistic manner with C6-ceramide to disrupt HDAC6/protein phosphatase 1 (PP1)/tubulin complex, to induce α-tubulin hyperacetylation, and to release and activate PP1, which then leads to AKT dephosphorylation and eventually causes cancer cell death. Interestingly, TSA itself results in short-term ceramide accumulation, which as a result of metabolic (glycosylation) removal, does not result in evident increase of cancer cell death. However, adding C6-ceramide led to a very pronounced increase in ceramide level and marked increase in cell death. Importantly, the effective synergistic anti-tumor activity of TSA plus C6-ceramide is also seen in in vivo mice xenograft pancreatic and ovarian cancer models, indicating that this regimen (HDACI plus C6-ceramide) may represent a more effective form of therapy against pancreatic and ovarian carcinoma.

The effects of a fat- and sugar-enriched diet and chronic stress on nonalcoholic fatty liver disease in male Wistar rats.

PURPOSE: The pathogenesis of nonalcoholic fatty liver disease (NAFLD) is still under debate. The aim of this study was to investigate the effects of a long-term fat- and sugar-enriched diet (FSED) and chronic stress (CS) on NAFLD. METHODS: Male Wistar rats were fed on either a standard diet or a FSED and given CS, a random electric foot shock (2 hr/morning and afternoon per day), or not for 12 weeks. After the experimental period, epididymal adipose tissue weight, sign of visceral obesity (VO), and hepatic index (HI) were measured. At sacrifice blood samples and liver were obtained. Histology of the liver was blindly determined by a pathologist. RESULTS: Histopathologically, moderate to severe steatosis, ballooning hepatocytes, and portal or lobules inflammation were observed in the FSED+CS group. However, mild to moderate steatosis with a few portal inflammation in the FSED group and mild steatosis or not with a few portal inflammation in the CS group were found correspondingly. In addition, more severe blood-fat disorder, high HI, fatty metabolic dysfunction, oxidative stress, high expressions of C-reactive protein mRNA and low expressions of peroxisome proliferator-activated receptor alpha mRNA in the liver were also revealed in the FSED+CS group. But, the degree of VO was not different between the FSED and FSED+CS groups. CONCLUSION: The observations strongly suggest that chronic stress can aggravate fat- and sugar-enriched diet-induced NAFLD from steatosis to steatohepatitis in male Wistar rats, although VO is not changed.

A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China.

There is a worldwide interest in studying SLC26A4 mutations that are responsible for enlarged vestibular aqueduct (EVA) in different ethnic background and populations. The spectrum of SLC26A4 mutations in Chinese population is yet to be fully characterized. In this study, all the 21 exons of SLC26A4 were screened in 107 Chinese patients with hearing loss associated with EVA or both EVA and Mondini dysplasia (MD), taken from six multiplex and 95 simplex families. The two types of control populations consisted of 84 normal-hearing subjects and 46 sensorineural hearing loss subjects without inner ear malformations. Biallelic mutations were found in 12 patients from multiplex families and 84 patients (88.4%) from the simplex families. In addition, monoallelic variant was detected in nine patients in the remaining 11 simplex families. Overall, up to 97.9% patients were found having at least one possible pathogenic variant in SLC26A4, with most having biallelic variants consistent with recessive inheritance of this disorder. A total of 40 mutations including 25 novel mutations were identified in the Chinese patients but were not detected in all the controls except for one normal subject. For the Chinese mutation spectrum of SLC26A4 gene, IVS 7-2A>G mutation was the most common form accounting for 57.63% (102/177) of all the mutant alleles.