Down-regulated microRNA-199a-3p enhances osteogenic differentiation of bone marrow mesenchymal stem cells by targeting Kdm3a in ovariectomized rats.

Osteoporosis is a prevalent systemic skeletal disorder entailing bone fragility and increased fracture risk, often emerging in post-menopausal life. Emerging evidence implicates the dysregulation of microRNAs (miRNAs or miRs) in the progression of osteoporosis. This study investigated the effect of miR-199a-3p on osteoporosis and its underlying mechanism. We first examplished an ovariectomized (OVX)-induced rat osteoporosis model, and then isolated mesenchymal stem cells (MSCs) from bone marrow of the model rats. The overexpression and knock down of miR-199a-3p were conducted in OVX rats and MSCs to verify the role of miR-199a-3p on MSC differentiation. Calcium nodules were measured using alizarin red S (ARS) staining. RT-qPCR and Western blot assay were performed to measure the expression of miR-199a-3p, Kdm3a and osteogenic differentiation-related markers in rat tissues and cells. The correlation between miR-199a-3p and Kdm3a was confirmed using dual-luciferase reporter assay. The enrichment of Kdm3a at the Erk2 and Klf2 promoter was assessed using chromatin immunoprecipitation (ChIP) assay. Isolated MSCs were positive for CD29, CD44, CD90, and CD45, suggesting successful isolation of MSCs. There was increased expression of miR-199a-3p and inhibited osteogenic differentiation in OVX rats. Kdm3a was negatively targeted by miR-199a-3p. Our results also demonstrated that Kdm3a elevated the expression of Erk2 and Erk2 by promoting Erk2 and Klf2 demethylation, which further contributed to osteogenic differentiation. Overall, our results revealed a regulatory network of miR-199a-3p in osteogenic differentiation, highlighting miR-199a-3p as a potential target for therapeutic interventions in osteoporosis.

miRTarBase 2020: updates to the experimentally validated microRNA-target interaction database.

MicroRNAs (miRNAs) are small non-coding RNAs (typically consisting of 18-25 nucleotides) that negatively control expression of target genes at the post-transcriptional level. Owing to the biological significance of miRNAs, miRTarBase was developed to provide comprehensive information on experimentally validated miRNA-target interactions (MTIs). To date, the database has accumulated >13,404 validated MTIs from 11,021 articles from manual curations. In this update, a text-mining system was incorporated to enhance the recognition of MTI-related articles by adopting a scoring system. In addition, a variety of biological databases were integrated to provide information on the regulatory network of miRNAs and its expression in blood. Not only targets of miRNAs but also regulators of miRNAs are provided to users for investigating the up- and downstream regulations of miRNAs. Moreover, the number of MTIs with high-throughput experimental evidence increased remarkably (validated by CLIP-seq technology). In conclusion, these improvements promote the miRTarBase as one of the most comprehensively annotated and experimentally validated miRNA-target interaction databases. The updated version of miRTarBase is now available at http://miRTarBase.cuhk.edu.cn/.

An innovative extraction strategy for herbal medicine by adopting p-sulphonatocalix[6]/[8]arenes.

INTRODUCTION: Alkaloids exist in various herbal medicine widely and exhibit diverse biological and pharmacological activities. p-Sulphonatocalix[6]arenes (SC6A) and p-sulphonatocalix[8]arenes (SC8A) are water-soluble supramolecular macrocycles and are applied to the extraction of alkaloids from herbal products. OBJECTIVE: In this study, an innovative method of SC6A/SC8A assisted extraction of the alkaloids from herbs was established. METHODS: SC6A and SC8A were designed to extract 27 alkaloids from seven herbal medicines. Based on the significant solubilisation and extraction effect, Stephaniae Tetrandrae Radix (Fangji, FJ) was selected to obtain the optimal extraction process by adopting single factor test and orthogonal experiment. Then, the alkaloids and SC6A/SC8A were separated by one-step alkalisation and SCnA were reused. The host-guest complexes between alkaloids and SCnA were determined by competitive fluorescence titration, differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR) and proton nuclear magnetic resonance (1 H-NMR) analysis. RESULTS: The optimum condition for SC6A assisted extraction was 5:1:80 (g/g/mL) for herbs/SC6A/solution ratio, 355-250 µm particle size and ultrasonicate 0.5 h, whilst 10:1:40 (g/g/mL) for herbs/SC8A/solution ratio, 355-250 µm particle size and ultrasonicate 0.5 h for SC8A assisted extraction. The total yield of alkaloids (fangchinoline and tetrandrine) from FJ was increased by 4.87 times and 5.97 times with SC6A and SC8A. Moreover, a good reusability of SC6A/SC8A was achieved by alkalisation dissociation. Host-guest complexes were determined by competitive fluorescence titration at a molar ratio of 1:1 between most alkaloids (25/27, except evodiamine and rutaecarpine) and SC6A/SC8A. The complex structure was proved by DSC, FTIR and 1 H-NMR analysis. CONCLUSION: The study provided an effective eco-friendly and energy-saving extraction method of alkaloids from herbal medicine.

[Study progress of supramolecular drug delivery system loading traditional Chinese medicine/natural anti-tumor products].

Ingredients with the anti-tumor property from traditional Chinese medicine/natural products have effects in treating tumor by regulating immune function, inhibiting tumor cell growth, inducing apoptosis, and reversing multidrug resistance. However, the potential of clinical application was restricted by the low hydrophilicity, stability and bioavailability and the lack of targeting. The above problems can be solved through research and development of supramolecular chemistry based on the non-covalent interactions between molecules. The supramolecular macrocyclic host molecules are characterized by a special cavity structure, low toxicity, high stability and rich derivatives spot. The anti-tumor supramolecular drug delivery system loading traditional Chinese medicine/natural products were designed to solve the above problems. In this paper, different supramolecular macrocyclic molecules, including crown ethers, cyclodextrin, calixarene, cucurbituril and two new-type supramolecular macrocyclic hosts were introduced. Their inclusion in enhancing drug solubility and stability, targeting drug delivery, synergies, combinative administration and new-type therapies in different applications, as well as such drug load modes as supramolecular polymers, indirect combination drug mode and polyrotaxanes were also reviewed in this paper, in order to provide a new idea for effective drug utilization.

Safety and efficacy of dehydrated ethanol soaking of the operative field in the treatment of spontaneous hepatocellular carcinoma rupture.

BACKGROUND: The aim of our study was to evaluate the clinical safety and value of ethanol surgical field infiltration (ESFI), combined with distilled water peritoneal lavage (DWPL), after hepatectomy in patients with hepatocellular carcinoma (HCC) rupture. METHODS: Rat liver tissue samples were soaked in dehydrated ethanol for different soaking times, and 18 rats were assigned to three groups that underwent different soaking methods of the hepatectomy cut surface. We retrospectively reviewed 45 patients who underwent hepatectomy for treatment of ruptured HCC. Among these, EFSI combined with DWPL was used in 21 patients (DAW group), with only DWPL used in the other 24 patients (DW group). Clinical outcomes were compared between the two groups. RESULTS: For in vitro experiments, the depth of coagulation degeneration and necrosis increased with the duration of soaking. For in vivo experiments, rats in all three groups survived until postoperative day 7 without significant postoperative complication. In patients, the rate of post-operation complication was comparable between the two groups (P = 0.398), with no between-group differences in liver function levels. The incidence of peritoneal dissemination was significantly higher for DW than DAW group (P = 0.037). Kaplan-Meier test identified dehydrated ethanol treatment as a significant factor of disease-free survival (DFS) (P = 0.036). On univariate analysis, dehydrated ethanol treatment was associated with better prognostic outcomes, although it was not retained as an independent factor of patient outcome. CONCLUSIONS: Dehydrated ethanol soaking of the cut surface of the hepatectomy could potentially lower the risk of metastasis and improve the effect of hepatectomy for ruptured HCC as well as showed potential therapeutic value for intraoperative iatrogenic rupture of HCC.

Isoflurane inhibits synaptic vesicle exocytosis through reduced Ca2+ influx, not Ca2+-exocytosis coupling.

Identifying presynaptic mechanisms of general anesthetics is critical to understanding their effects on synaptic transmission. We show that the volatile anesthetic isoflurane inhibits synaptic vesicle (SV) exocytosis at nerve terminals in dissociated rat hippocampal neurons through inhibition of presynaptic Ca(2+) influx without significantly altering the Ca(2+) sensitivity of SV exocytosis. A clinically relevant concentration of isoflurane (0.7 mM) inhibited changes in [Ca(2+)]i driven by single action potentials (APs) by 25 ± 3%, which in turn led to 62 ± 3% inhibition of single AP-triggered exocytosis at 4 mM extracellular Ca(2+) ([Ca(2+)]e). Lowering external Ca(2+) to match the isoflurane-induced reduction in Ca(2+) entry led to an equivalent reduction in exocytosis. These data thus indicate that anesthetic inhibition of neurotransmitter release from small SVs occurs primarily through reduced axon terminal Ca(2+) entry without significant direct effects on Ca(2+)-exocytosis coupling or on the SV fusion machinery. Isoflurane inhibition of exocytosis and Ca(2+) influx was greater in glutamatergic compared with GABAergic nerve terminals, consistent with selective inhibition of excitatory synaptic transmission. Such alteration in the balance of excitatory to inhibitory transmission could mediate reduced neuronal interactions and network-selective effects observed in the anesthetized central nervous system.

A prospective comparative study of hip resurfacing arthroplasty and large-diameter head metal-on-metal total hip arthroplasty in younger patients-a minimum of five year follow-up.

INTRODUCTION: Both hip resurfacing arthroplasty (HRA) and large-diameter head metal-on-metal total hip arthroplasty (LDH MoM THA) are generally used for young and active patients. A number of comparative studies of HRA and total hip arthroplasty have been published in the literature. However, studies that have compared HRA with LDH MoM THA are rare. The purpose of this study is to compare the mid-term results of HRA with those of LDH MoM THA in young patients. PATIENTS AND METHODS: Between 2007 and 2011, 68 patients were enrolled in the study and randomized into two groups: HRA group (28 hips) and LDH MoM THA group (40 hips). Peri-operative data including blood loss, surgery duration, size of the implant, and post-operative complications were recorded. All patients were assessed clinically and radiologically at six weeks; one, three and five years; and at the time of final review. Functional outcome were assessed using Harris hip (HHS), University of California Los Angeles (UCLA) and Oxford hip (OHS) scores. The mean follow-up for all patients was 7.4 years (5 to 9). RESULTS: Patient groups matched similarly in age, percent female, body mass index, preoperative HHS, and follow-up time. No differences were observed between the two groups in blood loss or in head size or acetabular inclination angle. HRA group had significantly longer surgery duration but less blood loss. The two groups had comparable HHS, UCLA, and OHS at the latest follow-up. Major complications, such as fracture, dislocation, infection, and adverse reactions to the metal debris (ARMD) were not found in the two groups. Only one case in LDH MoM THA group underwent revision surgery due to unexplained pain. CONCLUSION: Comparison of HRA and LDH MoM THA shows similar mid-term clinical results. HRA may be preferable due to the well-preserved bone stock and restoration of the native anatomy. LDH MoM THA may be used with caution due to the excessive metal ion release.

Transcriptome changes during TNF-α promoted osteogenic differentiation of dental pulp stem cells (DPSCs).

Dental pulp stem cells (DPSCs), due to the ease of isolation and their capacities of multi-lineage differentiation, are considered as attractive resources for regenerative medicine. In a previous study, we showed that TNF-α promoted the osteogenic differentiation of DPSCs via the NF-κB signaling pathway. However, the mechanisms of such differentiation were largely unknown. Here, we examined the gene expression profiles between undifferentiated, partially differentiated and fully differentiated DPSCs induced by TNF-α by performing the next-generation sequencing technique (RNA-Seq). Our results revealed a continuous transition of the transcriptome changes during TNF-α promoted osteogenic differentiation of DPSC. Bioinformatics analysis revealed a relatively general to specific transformation of the involved signaling pathways from the early to late stages of differentiation. Gene regulatory network analysis highlighted novel, key genes that are essential for osteogenic differentiation at different time points. These results were further validated by quantitative RT-PCR, confirming the high reliability of the RNA-Seq. Our data therefore will not only provide novel insights into the molecular mechanisms that drive the osteogenic differentiation of DPSCs, but also promote the studies of bone tissue engineering that utilizes DPSCs as a crucial resource.

α2-Adrenergic Receptor and Isoflurane Modulation of Presynaptic Ca2+ Influx and Exocytosis in Hippocampal Neurons.

BACKGROUND: Evidence indicates that the anesthetic-sparing effects of α2-adrenergic receptor (AR) agonists involve α2A-AR heteroreceptors on nonadrenergic neurons. Since volatile anesthetics inhibit neurotransmitter release by reducing synaptic vesicle (SV) exocytosis, the authors hypothesized that α2-AR agonists inhibit nonadrenergic SV exocytosis and thereby potentiate presynaptic inhibition of exocytosis by isoflurane. METHODS: Quantitative imaging of fluorescent biosensors of action potential-evoked SV exocytosis (synaptophysin-pHluorin) and Ca influx (GCaMP6) were used to characterize presynaptic actions of the clinically used α2-AR agonists dexmedetomidine and clonidine, and their interaction with isoflurane, in cultured rat hippocampal neurons. RESULTS: Dexmedetomidine (0.1 µM, n = 10) or clonidine (0.5 µM, n = 8) inhibited action potential-evoked exocytosis (54 ± 5% and 59 ± 8% of control, respectively; P < 0.001). Effects on exocytosis were blocked by the subtype-nonselective α2-AR antagonist atipamezole or the α2A-AR-selective antagonist BRL 44408 but not by the α2C-AR-selective antagonist JP 1302. Dexmedetomidine inhibited exocytosis and presynaptic Ca influx without affecting Ca coupling to exocytosis, consistent with an effect upstream of Ca-exocytosis coupling. Exocytosis coupled to both N-type and P/Q-type Ca channels was inhibited by dexmedetomidine or clonidine. Dexmedetomidine potentiated inhibition of exocytosis by 0.7 mM isoflurane (to 42 ± 5%, compared to 63 ± 8% for isoflurane alone; P < 0.05). CONCLUSIONS: Hippocampal SV exocytosis is inhibited by α2A-AR activation in proportion to reduced Ca entry. These effects are additive with those of isoflurane, consistent with a role for α2A-AR presynaptic heteroreceptor inhibition of nonadrenergic synaptic transmission in the anesthetic-sparing effects of α2A-AR agonists.

Gender differences of venous thromboembolism risk after total hip and total knee arthroplasty: a meta-analysis.

We performed a meta-analysis to evaluate gender differences of venous thromboembolism (VTE) risk after total hip (THA) and total knee arthroplasty (TKA). We searched PubMed and Web of Knowledge from their beginning to 25 July 2015. Pooled odds ratio (OR) and 95 % confidence interval (CI) for VTE risk were calculated. Twenty studies with 7,892,585 patients were included in our study. The VTE incidence ranged from 0.27 to 61.0 %. The sex ratio (male/female) was 0.623 (3,016,648/4,839,785) in no VTE group versus 0.492 (11,926/24,226) in VTE group. The pooled OR was 1.184 (95 % CI 1.070-1.310; Z = 3.28, P = 0.001). The Begg's test (z = 1.46, P = 0.144) and the Egger's test (t = 0.58, P = 0.571), and the funnel plot suggested there was no significant publication bias. Sensitivity analysis by omitted a study with largest simple size showed the pooled OR was 1.166 (95 % CI 1.051-1.294; Z = 2.91, P = 0.004) by random-effects model. Meta-regression showed VTE risk was not related with THA and TKA incidence (t = 0.35, P = 0.732). Our meta-analysis showed female patients have slightly higher risk of VTE than male patients after THA and TKA.

Prevention of Surgical Site Infection After Ankle Surgery Using Vacuum-Assisted Closure Therapy in High-Risk Patients With Diabetes.

Patients with diabetes have a high risk of surgical site infection (SSI) after ankle surgery. The aim of the present study was to investigate the efficacy of vacuum-assisted closure (VAC) in the prevention of SSI after ankle surgery compared with the efficacy of standard moist wound care (SMWC). A retrospective study was performed of unstable ankle fractures for surgical fixation in patients with diabetes from January 2012 to December 2014. VAC and SMWC were used for surgical incision coverage. The primary outcome was the incidence of SSI, and the secondary outcomes were the length of hospital stay and crude hospital costs. The data from 76 patients were analyzed, with 22 (28.95%) in the VAC group and 54 (71.05%) in the SMWC group. The incidence of SSI was 4.6% in the VAC group compared with 27.8% in the SMWC group (chi-square 5.076; p = .024), and the crude odds ratio for SSI in the VAC group was 0.124 (95% confidence interval 0.002 to 0.938). The length of hospital stay was lower in the VAC group than in the SMWC group (12.6 ± 2.7 days and 15.2 ± 3.5 days, respectively; t = 3.122, p = .003). The crude hospital costs were also lower in the VAC group than in the SMWC group (Chinese yuan 8643.2 ± 1195.3 and 9456.2 ± 1106.3, respectively; t = 2.839, p = .006). After logistic regression analysis, the adjusted odds ratio for the total SSI rate comparing VAC and SMWC was 0.324 (95% confidence interval 0.092 to 0.804; p = .021). Compared with SMWC, VAC can decrease the SSI rate after ankle surgery in patients with diabetes. This finding should be confirmed by prospective, randomized controlled clinical trials.

Angelica sinensis polysaccharides promotes apoptosis in human breast cancer cells via CREB-regulated caspase-3 activation.

Angelica sinensis polysaccharide (ASP) is purified from the fresh roots of Angelica sinensis (AS). This traditional Chinese medicine has been used for thousands of years for treating gynecological diseases and used in functional foods for the prevention and treatment of various diseases, such as inflammation and cancer. The antitumor activity of ASP is related to its biological activities, because it suppresses a variety of pro-proliferative or anti-apoptotic factors that are dramatically expressed in cancer cells of given types. In this study, we show that angelica sinensis polysaccharide induced apoptosis in breast cancer cells of T47D over-expressing the Cyclic AMP response element binding protein (CREB), inducing apoptosis-related signaling pathway activity. The result also found that ASP caused cell death was linked to caspase activity, accompanied by the loss of mitochondrial membrane potential, cytochrome c release, and Bax translocation from the cytosol to the mitochondria. We found that ASP significantly affected the poly-ADP-ribose polymerase (PARP), Bcl-2 Associated X Protein (Bax), Bcl-2, Bcl-xL and apoptotic protease activating facter-1 (Apaf1) protein expression in a dose- and time-dependent manner. DAPI staining and Flow cytometry were used to analyze apoptosis. The nude mice xenograft model was used to evaluate the antitumor effect of ASP in vivo. ASP has profound antitumor effect on T47D cells, probably by inducing apoptosis through CREB signaling pathway. Thus, these results suggest that ASP would be a promising therapeutic agent for breast cancer.

Astrocytic tumour grading: a comparative study of three-dimensional pseudocontinuous arterial spin labelling, dynamic susceptibility contrast-enhanced perfusion-weighted imaging, and diffusion-weighted imaging.

OBJECTIVES: We hypothesized that three-dimensional pseudocontinuous arterial spin labelling (pCASL) may have similar efficacy in astrocytic tumour grading as dynamic susceptibility contrast-enhanced perfusion-weighted imaging (DSC-PWI), and the grading accuracy may be further improved when combined with apparent diffusion coefficient (ADC) values. METHODS: Forty-three patients with astrocytic tumours were studied using diffusion weighted imaging (DWI), pCASL, and DSC-PWI. Histograms of ADC and normalized tumour cerebral blood flow values (nCBF on pCASL and nrCBF on DSC-PWI) were measured and analyzed. RESULTS: The mean 10 % ADC value was the DWI parameter that provided the best differentiation between low-grade astrocytoma (LGA) and high-grade astrocytoma (HGA). The nCBF and nrCBF (1.810 ± 0.979 and 2.070 ± 1.048) in LGA were significantly lower than those (4.505 ± 2.270 and 5.922 ± 2.630) in HGA. For differentiation between LGA and HGA, the cutoff values of 0.764 × 10(-3) mm(2)/s for mean 10 % ADC, 2.374 for nCBF, and 3.464 for nrCBF provided the optimal accuracy (74.4 %, 86.1 %, and 88.6 %, respectively). Combining the ADC values with nCBF or nrCBF could further improve the grading accuracy to 97.7 % or 95.3 %, respectively. CONCLUSIONS: pCASL is an alternative to DSC-PWI for astrocytic tumour grading. The combination of DWI and contrast-free pCASL offers a valuable choice in patients with risk factors. KEY POINTS: • pCASL shows positive correlation with DSC-PWI in astrocytic tumour grading. • ADC values based on ADC histograms can be an objective method. • Combination of DWI and pCASL or DSC-PWI can improve grading accuracy.

Hyperactive external awareness against hypoactive internal awareness in disorders of consciousness using resting-state functional MRI: highlighting the involvement of visuo-motor modulation.

Resting-state functional MRI (fMRI) has emerged as a valuable tool to characterize the complex states encompassing disorders of consciousness (DOC). Awareness appears to comprise two coexistent, anticorrelated components named the external and internal awareness networks. The present study hypothesizes that DOC interrupts the balance between the internal and external awareness networks. To gain more understanding of this phenomenon, the present study analyzed resting-state fMRI data from 12 patients with DOC versus 12 healthy age-matched controls. The data were explored using independent component analysis and amplitude of low-frequency fluctuation (ALFF) analysis. The results indicated that DOC deactivated midline areas associated with internal awareness. In addition, external awareness was strengthened in DOC because of increased activation in the insula, lingual gyrus, paracentral and supplementary motor area. The activity patterns suggested strengthened external awareness against weakened internal awareness in DOC. In particular, increased activity found in the insula, lingual gyrus, paracentral and supplementary motor area of patients with DOC implied possible involvement of augmented visuo-motor modulation in these patients. DOC is probably related to hyperactive external awareness opposing hypoactive internal awareness. This unique pattern of brain activity may potentially be a prognostic marker for DOC.

Association between type 2 diabetes mellitus and body composition based on MRI fat fraction mapping.

Purpose: To explore the association between type 2 diabetes mellitus (T2DM) and body composition based on magnetic resonance fat fraction (FF) mapping. Methods: A total of 341 subjects, who underwent abdominal MRI examination with FF mapping were enrolled in this study, including 68 T2DM patients and 273 non-T2DM patients. The FFs and areas of visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT) and abdominal muscle (AM) were measured at the level of the L1-L2 vertebral. The FF of bone marrow adipose tissue (BMAT) was determined by the averaged FF values measured at the level of T12 and L1 vertebral, respectively. The whole hepatic fat fraction (HFF) and pancreatic fat fraction (PFF) were measured based on 3D semi-automatic segmentation on the FF mapping. All data were analyzed by GraphPad Prism and MedCalc. Results: VAT area, VAT FF, HFF, PFF of T2DM group were higher than those of non-T2DM group after adjusting for age and sex (P < 0.05). However, there was no differences in SAT area, SAT FF, BMAT FF, AM area and AM FF between the two groups (P > 0.05). VAT area and PFF were independent risk factors of T2DM (all P < 0.05). The area under the curve (AUC) of the receiver operating characteristic (ROC) for VAT area and PFF in differentiating between T2DM and non-T2DM were 0.685 and 0.787, respectively, and the AUC of PFF was higher than VAT area (P < 0.05). Additionally, in seemingly healthy individuals, the SAT area, VAT area, and AM area were found to be significantly associated with being overweight and/or obese (BMI ≥ 25) (all P < 0.05). Conclusions: In this study, it was found that there were significant associations between T2DM and VAT area, VAT FF, HFF and PFF. In addition, VAT area and PFF were the independent risk factors of T2DM. Especially, PFF showed a high diagnostic performance in discrimination between T2DM and non-T2DM. These findings may highlight the crucial role of PFF in the pathophysiology of T2DM, and it might be served as a potential imaging biomarker of the prevention and treatment of T2DM. Additionally, in individuals without diabetes, focusing on SAT area, VAT area and AM area may help identify potential health risks and provide a basis for targeted weight management and prevention measures.

Altered neurotransmitter release, vesicle recycling and presynaptic structure in the pilocarpine model of temporal lobe epilepsy.

In searching for persistent seizure-induced alterations in brain function that might be causally related to epilepsy, presynaptic transmitter release has relatively been neglected. To measure directly the long-term effects of pilocarpine-induced status epilepticus on vesicular release and recycling in hippocampal mossy fibre presynaptic boutons, we used (i) two-photon imaging of FM1-43 vesicular release in rat hippocampal slices; and (ii) transgenic mice expressing the genetically encoded pH-sensitive fluorescent reporter synaptopHluorin preferentially at glutamatergic synapses. In this study we found that, 1-2 months after pilocarpine-induced status epilepticus, there were significant increases in mossy fibre bouton size, faster rates of action potential-driven vesicular release and endocytosis. We also analysed the ultrastructure of rat mossy fibre boutons using transmission electron microscopy. Pilocarpine-induced status epilepticus led to a significant increase in the number of release sites, active zone length, postsynaptic density area and number of vesicles in the readily releasable and recycling pools, all correlated with increased release probability. Our data show that presynaptic release machinery is persistently altered in structure and function by status epilepticus, which could contribute to the development of the chronic epileptic state and may represent a potential new target for antiepileptic therapies.

Internal fixation of intertrochanteric hip fractures: a clinical comparison of two implant designs.

OBJECTIVE: To compare two internal fixation devices clinically in stabilisation of intertrochanteric femur fractures. METHODS: Eighty-seven patients were randomised upon their admission to the hospital using a sealed envelope method. Forty-five were treated with proximal femur nail antirotation (PFNA) and 42 with reverse less invasive stabilisation system (LISS). The perioperative data were recorded and compared in relation to fracture type. RESULTS: In each type of fractures, no significant differences were found with respect to the blood loss, the quality of reduction, the time to bony healing, and the Harris hip score between the 2 groups. The mean duration of surgery was significantly longer in reverse LISS group than in PFNA group. CONCLUSION: Both the PFNA and the reversed LISS are effective in the treatment of different types of intertrochanteric femur fractures. PFNA is superior to reverse LISS in terms of surgical time, weight-bearing, and perhaps fluoroscopy time.

[Myocardial Slit2/Robo4 expression and impact of exogenous Slit2 on proliferation and migration of cardiac microvascular endothelial cells].

OBJECTIVE: To detect expression of Slit2 and Robo4 in mouse ventricular muscle blood vessel and explore the impact of exogenous Slit2 on proliferation and migrate of mouse cardiac microvascular endothelial cells. METHODS: Slit2 and Robo4 expression in mouse ventricular muscle blood vessel was detected by immunohistochemistry. Slit2 and Robo4 expression in cardiac microvascular endothelial cells isolated from mouse ventricular muscle were detected by euzymelinked immunosorbent assay and immunofluorescence, respectively. The effects of various concentrations exogenous Slit2 on proliferation of mouse cardiac microvascular endothelial cells was examined by CCK-8 cell proliferation kit. Transwell chamber was used to detect migration of mouse cardiac microvascular endothelial cells treated with 800 µl M199 culture medium containing 20%FBS (negative control), 10 ng/ml VEGF(positive control), 100 ng/ml Slit2(Slit2) and 100 ng/ml Slit2+10 ng/ml VEGF (Slit2+VEGF) and incubated for 18 h at 37 °C and 5%CO(2). RESULTS: Both Slit2 and Robo4 protein expressions were detected in ventricular muscle blood vessel. Slit2 protein expression was detected in mouse microvascular endothelial cells. Protein and mRNA Robo4 expressions were also evidenced in mouse microvascular endothelial cells. Proliferation of mouse cardiac microvascular endothelial cells was not affected by exogenous Slit2. Migration of mouse cardiac microvascular endothelial cells was not affected by exogenous Slit2 (22.1 ± 2.8 vs. 23.2 ± 3.8 in negative control, P > 0.05) and significantly enhanced by VEGF (65.3 ± 3.8, P < 0.05 vs. Slip2 and negative control), this effect could be blocked by cotreatment with Slip2 (29.2 ± 3.4 in Slip2+VEGF, P < 0.05 vs. VEGF) CONCLUSION: Slit2 and Robo4 are expressed in mouse ventricular muscle blood vessels and cardiac microvascular endothelial cells. Exogenous Slit2 has no impact on the proliferation of mouse cardiac microvascular endothelial cells but could inhibit VEGF-induced mouse cardiac microvascular endothelial cell migration.

A Comprehensive Quality Evaluation of Cimicifugae Rhizoma Using UPLC-Q-Orbitrap-MS/MS Coupled with Multivariate Chemometric Methods.

BACKGROUND: Cimicifugae Rhizoma, known in Chinese as Shengma, is a common medicinal material in traditional Chinese medicine (TCM), mainly used for treating wind-heat headaches, sore throat, uterine prolapse, and other diseases. OBJECTIVES: An approach using a combination of ultra-performance liquid chromatography (UPLC), MS, and multivariate chemometric methods was designed to assess the quality of Cimicifugae Rhizoma. METHODS: All materials were crushed into powder and the powdered sample was dissolved in 70% aqueous methanol for sonication. Chemometric methods, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least-squares discriminant analysis (OPLS-DA), were adopted to classify and perform a comprehensive visualization study of Cimicifugae Rhizoma. The unsupervised recognition models of HCA and PCA obtained a preliminary classification and provided a basis for classification. In addition, we constructed a supervised OPLS-DA model and established a prediction set to further validate the explanatory power of the model for the variables and unknown samples. RESULTS: Exploratory research found that the samples were divided into two groups, and the differences were related to appearance traits. The correct classification of the prediction set also demonstrated a strong predictive ability of the models for new samples. Subsequently, six chemical makers were characterized by UPLC-Q-Orbitrap-MS/MS, and the content of four components was determined. The results of the content determination revealed the distribution of representative chemical markers caffeic acid, ferulic acid, isoferulic acid, and cimifugin in two classes of samples. CONCLUSIONS: This strategy can provide a reference for assessing the quality of Cimicifugae Rhizoma, which is significant for the clinical practice and QC of Cimicifugae Rhizoma. HIGHLIGHTS: The HCA, PCA and OPLS-DA models visually classify Cimicifugae Rhizoma by appearance traits and obtain the chemical markers that influence the classification. The training and prediction sets were built to demonstrate the accuracy of the classification. Advanced UPLC-Q-Orbitrap-MS/MS technology provides powerful elucidation of critical chemical markers.

Enhancing the differentiation of walking and standing via the ratio of plantar pressures.

Differentiation of standing and walking based on plantar pressures is helpful in developing strategies to reduce health risks in the workplace. In order to improve the differentiation ability, the paper proposes a new metric for posture differentiation, that is, the pressure ratio on the two anatomical plantar regions. The plantar pressures were collected from 30 persons during walking and standing. After verifying the normal distribution of the pressure ratio by the Monte Carlo method, two-way repeated-measures ANOVA was conducted for the pressure ratios. The advantage of the pressure ratio over two conventional pressure metrics (the average pressure and the peak pressure) is demonstrated by its much larger size effect. Furthermore, the pressure ratio permits to establish value ranges corresponding to walking and standing, which are less influenced by specific person factors, thus facilitating the design of a standardized posture recognition system. The underlying mechanism underlying the pressure ratio is discussed from the aspect of biomechanics of movement.