The spatiotemporal matching pattern of Ezrin/Periaxin involved in myoblast differentiation and fusion and Charcot-Marie-Tooth disease-associated muscle atrophy.

BACKGROUND: Clinically, Charcot-Marie-Tooth disease (CMT)-associated muscle atrophy still lacks effective treatment. Deletion and mutation of L-periaxin can be involved in CMT type 4F (CMT4F) by destroying the myelin sheath form, which may be related to the inhibitory role of Ezrin in the self-association of L-periaxin. However, it is still unknown whether L-periaxin and Ezrin are independently or interactively involved in the process of muscle atrophy by affecting the function of muscle satellite cells. METHOD: A gastrocnemius muscle atrophy model was prepared to mimic CMT4F and its associated muscle atrophy by mechanical clamping of the peroneal nerve. Differentiating C2C12 myoblast cells were treated with adenovirus-mediated overexpression or knockdown of Ezrin. Then, overexpression of L-periaxin and NFATc1/c2 or knockdown of L-periaxin and NFATc3/c4 mediated by adenovirus vectors were used to confirm their role in Ezrin-mediated myoblast differentiation, myotube formation and gastrocnemius muscle repair in a peroneal nerve injury model. RNA-seq, real-time PCR, immunofluorescence staining and Western blot were used in the above observation. RESULTS: For the first time, instantaneous L-periaxin expression was highest on the 6th day, while Ezrin expression peaked on the 4th day during myoblast differentiation/fusion in vitro. In vivo transduction of adenovirus vectors carrying Ezrin, but not Periaxin, into the gastrocnemius muscle in a peroneal nerve injury model increased the numbers of muscle myosin heavy chain (MyHC) I and II type myofibers, reducing muscle atrophy and fibrosis. Local muscle injection of overexpressed Ezrin combined with incubation of knockdown L-periaxin within the injured peroneal nerve or injection of knockdown L-periaxin into peroneal nerve-injured gastrocnemius muscle not only increased the number of muscle fibers but also recovered their size to a relatively normal level in vivo. Overexpression of Ezrin promoted myoblast differentiation/fusion, inducing increased MyHC-I+ and MyHC-II + muscle fiber specialization, and the specific effects could be enhanced by the addition of adenovirus vectors for knockdown of L-periaxin by shRNA. Overexpression of L-periaxin did not alter the inhibitory effects on myoblast differentiation and fusion mediated by knockdown of Ezrin by shRNA in vitro but decreased myotube length and size. Mechanistically, overexpressing Ezrin did not alter protein kinase A gamma catalytic subunit (PKA-γ cat), protein kinase A I alpha regulatory subunit (PKA reg Iα) or PKA reg Iß levels but increased PKA-α cat and PKA reg II α levels, leading to a decreased ratio of PKA reg I/II. The PKA inhibitor H-89 remarkably abolished the effects of overexpressing-Ezrin on increased myoblast differentiation/fusion. In contrast, knockdown of Ezrin by shRNA significantly delayed myoblast differentiation/fusion accompanied by an increased PKA reg I/II ratio, and the inhibitory effects could be eliminated by the PKA reg activator N6-Bz-cAMP. Meanwhile, overexpressing Ezrin enhanced type I muscle fiber specialization, accompanied by an increase in NFATc2/c3 levels and a decrease in NFATc1 levels. Furthermore, overexpressing NFATc2 or knocking down NFATc3 reversed the inhibitory effects of Ezrin knockdown on myoblast differentiation/fusion. CONCLUSIONS: The spatiotemporal pattern of Ezrin/Periaxin expression was involved in the control of myoblast differentiation/fusion, myotube length and size, and myofiber specialization, which was related to the activated PKA-NFAT-MEF2C signaling pathway, providing a novel L-Periaxin/Ezrin joint strategy for the treatment of muscle atrophy induced by nerve injury, especially in CMT4F.

Continuous exposure to isoprenaline reduced myotube size by delaying myoblast differentiation and fusion through the NFAT-MEF2C signaling pathway.

We aimed to explore whether superfluous sympathetic activity affects myoblast differentiation, fusion, and myofiber types using a continuous single-dose isoprenaline exposure model in vitro and to further confirm the role of distinct NFATs in ISO-mediated effects. Compared with delivery of single and interval single, continuous single-dose ISO most obviously diminished myotube size while postponing myoblast differentiation/fusion in a time- and dose-dependent pattern, accompanied by an apparent decrease in nuclear NFATc1/c2 levels and a slight increase in nuclear NFATc3/c4 levels. Overexpression of NFATc1 or NFATc2, particularly NFATc1, markedly abolished the inhibitory effects of ISO on myoblast differentiation/fusion, myotube size and Myh7 expression, which was attributed to a remarkable increase in the nuclear NFATc1/c2 levels and a reduction in the nuclear NFATc4 levels and the associated increase in the numbers of MyoG and MEF2C positive nuclei within more than 3 nuclei myotubes, especially in MEF2C. Moreover, knockdown of NFATc3 by shRNA did not alter the inhibitory effect of ISO on myoblast differentiation/fusion or myotube size but partially recovered the expression of Myh7, which was related to the slightly increased nuclear levels of NFATc1/c2, MyoG and MEF2C. Knockdown of NFATc4 by shRNA prominently increased the number of MyHC +, MyoG or MEF2C + myoblast cells with 1 ~ 2 nuclei, causing fewer numbers and smaller myotube sizes. However, NFATc4 knockdown further deteriorated the effects of ISO on myoblast fusion and myotube size, with more than 5 nuclei and Myh1/2/4 expression, which was associated with a decrease in nuclear NFATc2/c3 levels. Therefore, ISO inhibited myoblast differentiation/fusion and myotube size through the NFAT-MyoG-MEF2C signaling pathway.

Nano-Pore Structure and Fractal Characteristics of Shale Gas Reservoirs: A Case Study of Longmaxi Formation in Southeastern Chongqing, China.

Pore structure and fractal dimensions can characterize the adsorption, desorption and seepage characteristics of shale gas reservoirs. In this study, pore structure, fractal characteristics and influencing factors were studied of the Longmaxi formation shale gas reservoir in southeastern Chongqing, China. Scanning electron microscopy was used to describe the characteristics of various reservoirs. High pressure mercury intrusion and low temperature liquid N2 and CO2 adsorption experiments were used to obtain pore structure parameters. V-S model, FHH model and Menger sponge model were selected to calculate the micropore, mesopore and macropore fractal dimensions, respectively. The results show that organic matter pores, inter-granular pores, intra-granular pores and micro-fractures are developed within the shale, and the pore morphology is mostly ink pores and parallel plate pores with aperture essentially in the 1-2 nm and 2-50 nm ranges. Moreover, macropores are the most complex in these samples, with mesopores being less complex than macropores, and the micropores being the simplest. D1 (micropore fractal dimension) ranges from 2.31 to 2.50, D2 (mesopore fractal dimension) ranges from 2.74 to 2.83, D3 (macropore fractal dimension) ranges from 2.87 to 2.95, and Dt (comprehensive fractal dimension) ranges from 2.69 to 2.83 of fractal characteristics. D1 and D2 are mainly controlled by TOC content, while D3 and Dt are mainly controlled by brittle and clay mineral content. These results may be helpful for exploration and the development of shale gas in southeastern Chongqing, China.

Hydrocarbon Generation and Chemical Structure Evolution from Confined Pyrolysis of Bituminous Coal.

The molecular composition of organic matter formed during pyrolysis is complex. Fourier transform infrared spectroscopy (FTIR) is a good technique to investigate the coal chemical structural evolution. However, reports on the effects of chemical structure on the n-alkane yields and their relative functional groups are scarce in the literature. In our case, the chemical structural evolution process of bituminous coal obtained by pyrolysis at two different heating rates has been analyzed by pyrolysis-gas chromatography (Py-GC) and FTIR. Furthermore, some of the small molecular compounds (e.g., n-alkanes 24 can generate n-alkanes 20 or low-weight compounds) generated by gold-tube pyrolysis were identified using other GC techniques. Biomarkers were analyzed and compared to generated n-alkanes from the gold-tube pyrolysis experiments. We present the results of the relationship between the FTIR parameters and the molecular compositions that were analyzed. A good linear relationship can be seen between the FTIR parameters (C=O, C=C, and C-factor values), the carbon preference index (CPI), and the ratio of the pristane content and n-C17 alkane content (Pr/n-C17). Furthermore, the n-alkane fraction of the pyrolysates, in particular pristane, phytane, n-C17 alkane, and n-C18 alkane, changed upon maturation. Our conclusions indicate that FTIR is applicable as a structural and chemical change probe to explore the pyrolysis process.

Rhythmic low-field magnetic stimulation may improve depression by increasing brain-derived neurotrophic factor.

BACKGROUND: Low-field magnetic stimulation (LFMS) has mood-elevating effect, and the increase of brain-derived neurotrophic factor (BDNF) is associated with antidepressant treatment. We evaluated the effects and association with BDNF of rhythmic LFMS in the treatment of major depressive disorder (MDD). METHODS: A total of 22 MDD patients were randomized to rhythmic alpha stimulation (RAS) or rhythmic delta stimulation (RDS), with 5 sessions per week, lasting for 6 weeks. Outcomes assessments included the 17-item Hamilton Depression Rating Scale (HAMD-17), the Hamilton Anxiety Rating Scale (HAMA), and the Clinical Global Impressions-Severity scale (CGI-S) at baseline and at weeks 1, 2, 3, 4, and 6. Serum BDNF level was measured at baseline and at weeks 2, 4, and 6. RESULTS: HAMD-17, HAMA, and CGI-S scores were significantly reduced with both RAS and RDS. RAS patients had numerically greater reductions in HAMD-17 scores than RDS patients (8.9 ± 7.4 vs. 6.2 ± 6.2, effect size [ES]=0.40), while RDS patients had greater improvement in HAMA scores (8.2 ± 8.0 vs. 5.3 ± 5.8, ES=0.42). RAS was associated with clinically relevant advantages in response (54.5% vs. 18.2%, number-needed-to-treat [NNT]=3) and remission (36.4% vs. 9.1%, NNT=4). BDNF increased significantly during the 6-week study period (p<0.05), with greater increases in RAS at weeks 4 and 6 (ES=0.66-0.76) and statistical superiority at week 2 (p=0.034, ES=1.23). Baseline BDNF in the 8 responders (24.8±9.0 ng/ml) was lower than in the 14 nonresponders (31.1±7.3 ng/ml, p=0.083, ES=-0.79), and BDNF increased more in responders (8.9±7.8 ng/ml) than in nonresponders (1.8±3.5 ng/ml, p=0.044). The change in BDNF at week 2 was the most strongly predicted response (p=0.016). CONCLUSIONS: Rhythmic LFMS was effective for MDD. BDNF may moderate/mediate the efficacy of LFMS.

Clinical efficacy, onset time and safety of bright light therapy in acute bipolar depression as an adjunctive therapy: A randomized controlled trial.

BACKGROUND: Bright light therapy (BLT) is an effective treatment for seasonal affective disorder and non- seasonal depression. The efficacy of BLT in treating patients with bipolar disorder is still unknown. AIMS: The aim of this study is to examine the efficacy, onset time and clinical safety of BLT in treating patients with acute bipolar depression as an adjunctive therapy (trial registration at ClinicalTrials.gov: NCT02009371). METHODS: This was a multi-center, single blind, randomized clinical trial. Seventy-four participants were randomized in one of two treatment conditions: BLT and control (dim red light therapy, dRLT). Sixty-three participants completed the study (33 BLT, 30 dRLT). Light therapy lasted for two weeks, one hour every morning. All participants were required to complete several scales assessments at baseline, and at the end of weeks 1 and 2. The primary outcome measures were the clinical efficacy of BLT which was assessed by the reduction rate of HAMD-17 scores, and the onset time of BLT which was assessed by the reduction rate of QIDS-SR16 scores. The secondary outcome measures were rates of switch into hypomania or mania and adverse events. RESULTS: 1) Clinical efficacy: BLT showed a greater ameliorative effect on bipolar depression than the control, with response rates of 78.19% vs. 43.33% respectively (p < 0.01). 2) Onset day: Median onset day was 4.33 days in BLT group. 3) BLT-emergent hypomania: No participants experienced symptoms of hypomania. 4) Side effects: No serious adverse events were reported. CONCLUSION: BLT can be considered as an effective and safe adjunctive treatment for patients with acute bipolar depression.

Adding dexmedetomidine to ropivacaine for lumbar plexus and sciatic nerve block for amputation of lower limb in high-risk patient-a case report.

The ischemia necrosis of limb frequently requires surgery of amputation. Lumbar plexus and sciatic nerve block is an ideal intra-operative anesthetic and post-operative antalgic technique for patients of amputation, especially for high-risk patients who have severe cardio-cerebrovascular diseases. However, the duration of analgesia of peripheral nerve block is hardly sufficient to avoid the postoperative pain and the usage of opioids. In this case, a 79-year-old man, with multiple cerebral infarcts, congestive heart failure, atrial flutter and syncope, was treated with an above knee amputation because of ischemia necrosis of his left lower limb. Dexmedetomidine 1 µg/kg was added to 0.33% ropivacaine for lumbar plexus and sciatic nerve block in this case for intra-operative anesthesia and post-operative analgesia. The sensory function was blocked fully for surgery and the duration of analgesia maintained 26 hours with haemodynamic stability and moderate sedation. The patient did not complain pain and require any supplementary analgesics after surgery. This case showed that adding 1 µg/kg dexmedetomidine to ropivacaine for lumbar plexus and sciatic nerve block may be a feasible and safe technique for high-risk patients for lower limb surgery of amputation.

Measurement-Based Care Versus Standard Care for Major Depression: A Randomized Controlled Trial With Blind Raters.

OBJECTIVE: The authors compared measurement-based care with standard treatment in major depression. METHODS: Outpatients with moderate to severe major depression were consecutively randomized to 24 weeks of either measurement-based care (guideline- and rating scale-based decisions; N=61), or standard treatment (clinicians' choice decisions; N=59). Pharmacotherapy was restricted to paroxetine (20-60 mg/day) or mirtazapine (15-45 mg/day) in both groups. Depressive symptoms were measured with the Hamilton Depression Rating Scale (HAM-D) and the Quick Inventory of Depressive Symptomatology-Self-Report (QIDS-SR). Time to response (a decrease of at least 50% in HAM-D score) and remission (a HAM-D score of 7 or less) were the primary endpoints. Outcomes were evaluated by raters blind to study protocol and treatment. RESULTS: Significantly more patients in the measurement-based care group than in the standard treatment group achieved response (86.9% compared with 62.7%) and remission (73.8% compared with 28.8%). Similarly, time to response and remission were significantly shorter with measurement-based care (for response, 5.6 weeks compared with 11.6 weeks, and for remission, 10.2 weeks compared with 19.2 weeks). HAM-D scores decreased significantly in both groups, but the reduction was significantly larger for the measurement-based care group (-17.8 compared with -13.6). The measurement-based care group had significantly more treatment adjustments (44 compared with 23) and higher antidepressant dosages from week 2 to week 24. Rates of study discontinuation, adverse effects, and concomitant medications did not differ between groups. CONCLUSIONS: The results demonstrate the feasibility and effectiveness of measurement-based care for outpatients with moderate to severe major depression, suggesting that this approach can be incorporated in the clinical care of patients with major depression.

Impurity profiling of etimicin sulfate by liquid chromatography ion-trap mass spectrometry.

A reversed phase (RP)-LC method using a C(18) column resistant to basic pH and an alkaline (pH 10) aqueous mobile phase was developed and coupled to MS with an electrospray ionization (ESI) source in the positive ion mode which provides MS(n) capability. In total, 26 impurities were detected in a commercial sample. The structures of the impurities were proposed based on comparison of their fragmentation patterns with those of the available reference substances, the synthetic route and literature data. Starting material and its residual impurities, intermediates, synthetic by-products and degradation products were the main sources of those impurities. 14 impurities described in this work were newly identified.

[Trabeculectomy with amniotic membrane transplantation and combining suture lysis of scleral flap in complicated glaucoma].

OBJECTIVE: To study and evaluate the outcome of trabeculectomy with amniotic membrane transplantation combining suture lysis of scleral flap in complicated glaucoma. METHODS: This operation was performed in 48 patients (67 eyes) with complicated glaucoma. Amniotic membrane was implanted under the scleral flap and the conjunctival flap and combined with suture lysis for scleral flap to control the outflow of the aqueous humor. RESULTS: Follow-up period ranged from 5 to 38 months (mean 19.5 months). The intraocular pressure (IOP) was lowered from (43.86 +/- 7.12) mm Hg (preoperative IOP) to postoperative (19.63 +/- 2.57) mm Hg (postoperative IOP) in neovascular glaucoma (t = 4.96, P < 0.001); from (40.31 +/- 4.79) mm Hg to (18.09 +/- 2.21) mm Hg in glaucoma after IOL implant (t = 3.54, P < 0.05); from (37.94 +/- 5.63) mm Hg to (20.14 +/- 3.15) mm Hg in aphakic glaucoma (t = 5.12, P < 0.05); from (32.48 +/- 3.98) mm Hg to (16.54 +/- 1.84) mm Hg in juvenile glaucoma (t = 4.23, P < 0.01); from (36.01 +/- 4.13) mm Hg to (18.11 +/- 3.40) mm Hg in uveitic glaucoma (t = 4.47, P < 0.01); from (34.43 +/- 5.28) mm Hg to (18.31 +/- 1.52) mm Hg in glaucoma after failure of conventional filtering operation (t = 2.05, P < 0.05). Statistical significant difference was found in the IOP before and after the operation. The rate of formation of functional bulb was 80.6%. There were no reject reaction and severe complications. CONCLUSION: Trabeculectomy with amniotic membrane transplantation combining suture lysis of scleral flap is a useful method in complicated glaucoma.