MicroRNA-22 targets FMNL2 to inhibit melanoma progression via the regulation of the Wnt/ß-catenin signaling pathway and epithelial-mesenchymal transition.

OBJECTIVE: Melanoma is regarded as one common malignancy in skin cancers, and there is growing evidence that microRNAs (miRNAs) play a vital role in the oncogenesis of tumors. This study aimed to investigate the roles and mechanism of miR-22 in melanoma. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was utilized to detect the expressions of miR-22 and mRNA. The functions of miR-22 in melanoma cell proliferation, migration and invasion were investigated with functional assays, including MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and transwell assay. Western blots were utilized to examine the protein expressions. Luciferase reporter analysis was conducted to confirm the interactions between formin-like 2 (FMNL2) and miR-22 in melanoma cells. FMNL2 expression levels in melanoma tissues were investigated by immunohistochemistry (IHC) assays. RESULTS: The qRT-PCR analysis demonstrated significant decreased miR-22 expressions in melanoma tissues. Decreased miR-22 in melanoma tissues were correlated with adverse clinicopathologic features and poor prognosis. Functional assays indicated that upregulation inhibited melanoma cell proliferation, invasion and migration capacities. Luciferase reporter assays showed that FMNL2 was targeted by miR-22 in melanoma cells. Western blots indicated that miR-22 exerted anti-tumor functions by regulating the Wnt/ß-catenin and epithelial-mesenchymal transition (EMT). CONCLUSIONS: Our findings showed that miR-22 served as a tumor suppressor in melanoma progression, implying that miR-22 may function as a novel therapeutic target and prognostic biomarker for melanoma treatments.

[Continuous fascia iliaca compartment block combined with oral analgesics for pre-operative pain control in elderly hip fracture patients].

Objective: The purpose of this study was to assess the efficacy of ultrasound-guided continuous fascia iliaca compartment block combined with oral analgesics for pre-operative pain control in elderly patients with hip fracture. Methods: One hundred and sixteen patients with hip fractures in Xuan Wu Hospital of Capital Medical University during Dec. 2015 to Dec. 2016 were included. These patients were randomized into 2 groups: control group (group C) (receiving the traditional analgesia: tramadol 50 mg and paracetamol 500 mg tid, po from admission to surgery), study group (group S) (receiving ultrasound-guided continuous fascia iliaca compartment block combined with oral analgesics for pre-operative pain control from admission to surgery). Pain relief or pain intensity was assessed preoperatively at before administration of analgesia (T0), 1 h after administration of analgesia (T1), the second day after admission(T2), in the morning of surgery day (before surgery) (T3) using a visual analog scale. The satisfaction score with the analgesic regimen preoperatively was recorded. The amount of rescue analgesia, occurrence of adverse events (nausea, vomiting, respiratory depression, over sedation) and continuous fascia iliaca compartment block complications were also collected. Results: Pain scores (VAS) at passive movement of group S were significantly lower at T1 (32±8), T2 (32±8) and T3 (34±7) than that at T0 (73±12) (all P<0.05). VAS at rest of group S were significantly lower than those of group C at T3 (t=3.488, P<0.05). VAS at passive movement of group S were significantly lower than those of group C at T1,T2,T3 (P<0.05). The satisfaction score with the analgesic regimen was greater in group S (74±10) than that in group C (46±11) (t=-14.209, P<0.05). The incidence of rescue analgesia was lower in group S (0) than in group C (17.2%) (χ2=5.472, P<0.05). The occurrence of nausea and vomiting was 6.9% and 1.7% in group S, which were lower than that in group C (22.4%, 12.1%) (χ2=6.779, 2.416, all P<0.05). There were no obvious complications of continuous fascia iliaca compartment block in group S. Conclusion: Ultrasound guided continuous fascia iliaca compartment block combined with oral analgesics preoperatively is an effective way of providing analgesia for elderly with hip fracture, which can improve the patient's comfort and satisfaction.

Reducing postsurgical exudate in breast cancer patients by using San Huang decoction to ameliorate inflammatory status: a prospective clinical trial.

Background: Reducing inflammatory factors in wound exudate is a promising treatment approach for healing wounds in postsurgical breast cancer patients. Traditional Chinese Medicine (tcm) treatments have been shown to be beneficial and safe for optimal regulation of oxidative stress during the postoperative period. In the present clinical trial, we evaluated the effectiveness of a promising Chinese herbal formula, San Huang decoction [shd (Radix astragali, Radix et rhizoma rhei, and Rhizoma curcuma longa, 3:1:1; supplemental Table 1)], on wound inflammatory response after mastectomy. Methods: The study randomized 30 patients with breast cancer who fulfilled the inclusion and exclusion criteria to either a treatment (n = 15) or a control group (n = 15). Patients in the treatment group received liquid shd, taken twice daily with or without food. Treatment was given for 1 day before surgery and for 7 days postoperatively. Participants in the control group received a placebo on the same schedule as the treatment group. Outcomes measured in every subject included clinical tcm and wound inflammation symptom scores, daily and total amounts of drainage fluid, and levels of inflammatory factors in the exudate [tumour necrosis factor α (tnf-α), interleukins 6 (il-6), 8 (il-8), and 2R (il-2R), human C-reactive protein (crp)] at 2 hours and on days 1, 3, and 7 postoperatively. Results: The total amount of drainage fluid over 7 days was significantly lower in the treatment group (572.20 ± 93.95 mL) than in the control group (700.40 ± 107.38 mL). The tcm symptom score was also lower in treatment group (day 7: 1.87 ± 0.83 vs. 4.80 ± 3.61, p = 0.049), as was the inflammatory symptom score (day 7: 0.67 ± 0.72 vs. 3.67 ± 2.50, p = 0.001). Levels of tnf-α, il-6, il-8, il-2R, and crp in drainage fluid were significantly lower with shd treatment. Conclusions: Perioperative treatment with shd effectively lessened postoperative exudate and ameliorated inflammatory symptoms in patients who underwent surgery for breast cancer.

Effects of Graded Levels of Isomaltooligosaccharides on the Performance, Immune Function and Intestinal Status of Weaned Pigs.

The objective of this study was to investigate the effects of graded levels of isomaltooligosaccharides (IMO) on the performance, immune function and intestinal microflora and intestinal mucosal morphology of weaned pigs. In a 28-day experiment, one hundred eighty, twenty eight-day-old, crossbred (Duroc×Large White×Landrace), weaned pigs, with an initial body weight of 8.19±1.45 kg, were fed either an unsupplemented corn-soybean meal based diet or similar diets supplemented with 0.2%, 0.4%, 0.6%, or 0.8% IMO added at the expense of corn. Each treatment was replicated six times with six pigs (three barrows and three gilts) per pen. From day 0 to 14, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p = 0.05) linearly declined as the IMO level increased. On d 14, the level of the immunoglobulins IgA, IgM, and IgG in the serum of pigs were linearly increased (p<0.05) with increasing IMO supplementation. Interleukin-6 (IL-6) was linearly (p<0.05) and quadratically (p<0.05) decreased as IMO intake increased. From day 15 to 28, there was a trend for weight gain to be linearly increased, and IL-2 was linearly (p<0.05) increased as IMO supplementation increased on d 28. Over the entire experiment, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p<0.05) was linearly decreased as the IMO level increased. Supplementation with IMO had no effect on the intestinal microflora of pigs in the ileum and cecum of pigs, as well as the villus height and crypt depth in the ileum and jejunum (p>0.05). These results indicate that dietary inclusion of IMO increases weight gain, gain:feed and enhanced the immune status of pigs, and could be a valuable feed additive for use in weaned pigs, particularly during the period immediately after weaning.

Effects of osteopontin on the expression of IL-6 and IL-8 inflammatory factors in human knee osteoarthritis chondrocytes.

OBJECTIVE: Osteoarthritis (OA) is a chronic musculoskeletal disease characterized by progressive destruction of articular cartilage, OA lead to chronic pain and functional restrictions in affected joints. The present study was to investigate the role of osteopontin (OPN) in the athogenesis of OA through studying the effect of OPN on expression of IL-6 and IL-8 inflammatory factors in human OA chondrocytes. PATIENTS AND METHODS: One-step type II collagenase digestive method was used to isolate OA chondrocytes from sectional cartilage specimens of 16 primary knee OA patients received total knee replacement surgery. Synchronized first-generation chondrocytes were then treated with OPN (100 ng/ml or 1 µg/mL). The changes in cell morphology of OA chondrocytes were analyzed before and after treated with OPN; and the expression levels of IL-6 and IL-8 were evaluated by real-time q-PCR. RESULTS: Chondrocytes were successfully isolated from human OA knee cartilage, and the viability of isolated chondrocytes was 92.11±3.13%. Adherent chondrocytes formed clusters of irregular polygonal shape with intercellular pseudopodia extension. After OPN treatment, cells became fusiform or irregularly shaped, and the number of intercellular pseudopodia decreased significantly. The mRNA expression of IL-6 increased to 1.83 times at 0.1 µg/ml of OPN and 3.1 times at the dose of 1 µg/ml; the expression of IL-8 increased to 1.57 and 3.27 times at the dose of 0.1 µg/ml and 1 µg/ml respectively. CONCLUSIONS: OPN could up-regulate expression of IL-6 and IL-8 cytokines in human OA chondrocytes, and the expression increased with the increasing concentration of OPN, which might be one of the potential mechanisms of OPN in the development of OA.

Transcutaneous electrical nerve stimulation on acupoints reduces fentanyl requirement for postoperative pain relief after total hip arthroplasty in elderly patients.

BACKGROUND: Transcutaneous electrical nerve stimulation (TENS) is regarded as an effective treatment for various types of pain. However, no randomized controlled trial has investigated TENS on acupoints for postoperative analgesia in elderly patients. This study aim to investigate whether TENS on acupoints has any favorable effect on complementary analgesia after total hip arthroplasty (THA) for elderly patients compared with a sham control treatment. METHODS: Sixty-eight elderly patients requiring THA surgery were enrolled and randomly allocated to one of two groups. Group Acu received true TENS on acupoints (bilateral P6, L14; ST36, GB31 ipsilateral to the surgery site) and Group Sham received sham treatment. All patients received patient-controlled analgesia for two days postoperatively. Analgesia was assessed by postoperative fentanyl requirement and pain intensity using a visual analogue scale (VAS-10 cm). The incidence of analgesia-related side effects, optional medication use and effects of patients' blinding were recorded. RESULTS: Fentanyl consumption in Group Acu was lower than that in Group Sham at 24 h (mean ± SD; 360±117 vs. 572±132 µg; P<0.001) and 48 h (712±184 vs. 1022±197 µg; P<0.001) after surgery. Postoperative pain intensity measured by VAS was similar in both groups. The incidence of opioid-related side effects and rescue medication for postoperative analgesia was significantly higher in Group Sham than in Group Acu. Differences between the groups regarding the effects of patients' blinding were not significant. CONCLUSION: TENS on specific acupoints is an effective and complementary approach to reduce postoperative analgesic requirement in elderly patients after THA.

Bilateral Bispectral Index (BIS)-Vista as a measure of physiologic sleep in sleep-deprived anesthesiologists.

BACKGROUND: Anesthesiologists who work extended hours exhibit acute and chronic sleep deprivation. The newest bilateral Bispectral Index (BIS)-Vista monitor offers new advantages over earlier models, which include a new algorithm (version 1.4). One possible useful application for BIS monitoring of physiological sleep would be in critically ill patients who almost uniformly suffer from sleep disruption and deprivation because they loose their circadian rhythm due to the harsh noisy environment of critical care units. Previous studies, which used earlier versions of the BIS monitor to depict normal physiologic sleep, produced widely conflicting results. The aim of our study was to assess whether the new BIS-Vista monitor would exhibit a temporal decline that corresponds to natural physiologic sleep stages in healthy sleep-deprived, post-call anesthesiologists. METHODS: BIS-Vista sensors were bilaterally mounted, according to the manufacturer guidelines, onto the forehead of 10 healthy sleep-deprived volunteer anesthesiologists. Using the conventional Rechtschaffen and Kales criteria, BIS values during the awake, S1, S2, S3, S4, and rapid eye movement (REM) sleep stages were recorded. RESULTS: Mean±SD BIS values during the awake, S1, S2, S3, S4, and REM sleep stages significantly declined for the left (96±2, 82±11, 73±10, 53±13, 43±11, 66±9) and right (97±2, 78±11, 69±9, 50±10, 39±13, 61±14) hemispheres respectively. There were no significant interhemispheric differences in BIS values over time (two-way ANOVA). However, in one subject, the left and right BIS values were mostly discordant throughout the recordings. CONCLUSION: We demonstrated that although the BIS-Vista monitor was neither designed nor validated for monitoring normal physiologic sleep, it depicted a temporal decline that corresponds to normal physiologic sleep stages in sleep-deprived anesthesiologists.

Autophagy inhibition contributes to radiation sensitization of esophageal squamous carcinoma cells.

Radiotherapy is a useful component of treatment strategies for esophageal cancer. The role of autophagy in response to ionizing radiation was investigated in human esophageal squamous carcinoma cells. Cell viability and clonogenic survival assay were used to evaluate the radiosensitivity of autophagy inhibitor (3-MA) on esophageal squamous carcinoma cells. The percentage of apoptotic cells and cell cycle analysis were assessed by flow cytometry; DAPI staining was used to detect apoptotic cells. The expression of beclin-1 and LC3 was measured using a Western blot. The ultrastructural analysis was under the electron microscope. 6 Gy irradiation induced a massive accumulation of autophagosomes accompanied by strong upregulation of beclin-1 and LC3-II expression in TE-1 cells. Compared with radiation alone, 3-MA combined with radiation significantly decreased cell viability, as well as autophagic ratio, beclin-1, and LC3-II protein level. Inhibition of autophagy increased radiation-induced apoptosis and the percentage of G2/M-phase cells. Blockade of autophagy with 3-MA enhanced cytotoxicity of radiotherapy in human esophageal squamous carcinoma cells. It suggests that inhibition of autophagy could be used as adjuvant therapy to treat esophageal squamous cell carcinoma.

qBrn-2, a POU III gene in quail: distinct developmental expression revealed by a specific antibody.

We examined qBrn-2 protein expression in quail from its onset to final profile with a specific antibody we prepared. qBrn-2 expression employed onset-widespread-restriction pattern, and precisely concurred with formation and differentiation of neural tube. qBrn-2 protein was also located outside neural tube. Obvious differences in expression were observed compared with that of Brn-2.

Isolation of quail qMEF2D gene and its expression pattern in the developing central nervous system.

We report here the identification of the first avian MEF2 gene, termed qMEF2D. qMEF2D is the first MEF2 protein that contains 41 repeats of glutamine in the C-terminal. This quail gene is more abundantly expressed, in a transient fashion, in the developing brain than in the muscle cells.

[Effects of the combination of Astragalus membranaceus (Fisch.) Bge. (AM), Angelica sinensis (Oliv.) Diels (TAS), Cyperus rotundus L. (CR), Ligusticum chuangxiong Hort (LC) and Peaonìa veitchii lynch (PV) on the hemorheological changes in "blood stagnating" rats].

The "blood stagnating" rat model was built with adrenaline and cold stimulation. Its hemorrheological character was an increase in the viscosity, thickness and liability to coagulate. The experimental result showed that AM and TAS could decrease the whole blood specific viscosity, but at the same time increase the plasma specific viscosity. The qi-regulating drug CR and two blood-activating drugs LC and PV could improve the hemorrheological changes in "blood stagnating" rats. The combination of qi-regulating drugs and blood-activating drugs had more favorable effect.

[Effects of the combination of Astragalus membranaceus (Fisch.) Bge. (AM), tail of Angelica sinensis (Oliv.) Diels. (TAS), Cyperus rotundus L. (CR), Ligusticum chuanxiong Hort. (LC) and Paeonia veitchii Lynch (PV) on the hemorrheological changes in normal rats].

The results showed that AM and TAS had significant effects of enriching the blood. CR, a Qi-regulating drug, LC and PV, two blood-activating drugs, could improve all hemorrheological indexes, such as the whole blood specific viscosity, the plasma specific viscosity, erythrocyte electrophoresis, etc. The combination of Qi-regulating drug and blood-activating drug displayed more favorable effect. This experiment has provided some pharmacological evidence for the theory of "Qi Xue Xiang Guan" (correlation of vital energy with blood circulation) in traditional Chinese medicine.