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Aims: To study the protective effects of late remote ischaemic preconditioning (RIPC) against myocardial ischaemia/reperfusion (I/R) injury and determine whether Stat5 is involved in this protection by using cardiomyocyte-specific Stat5 knockout mice (Stat5-cKO)., Methods and Results: Mice were exposed to lower limb RIPC or sham ischaemia. After 24 h, the left anterior descending artery (LAD) was ligated for 30 min, then reperfused for 180 min. The myocardial infarct size (IS), apoptotic rate of cardiomyocytes, and serum myocardial enzymes were measured to evaluate for cardioprotective effects. Heart tissues were harvested to determine the cardiomyocytes' anti-apoptotic and survival signaling. When compared with the Stat5fl/fl mice without RIPC, Stat5fl/fl mice with RIPC (Stat5fl/fl+RIPC + I/R) displayed a decreased myocardial IS/LV (16 ± 1.5 vs. 30.1 ± 3.1%, P < 0.01; IS/ area at risk (AAR), 42.2 ± 3.5 vs. 69.2 ± 4.9%, P < 0.01), a reduced cardiomyocyte apoptotic rate (2.1 ± 0.37 vs. 5.5 ± 0.53%, P < 0.01), and lower creatine kinase (CK), lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) levels. To the contrary, the Stat5-cKO mice (Stat5fl/fl; Tnnt2Cremice with Doxycycline treatment for 7 days) did not exhibit any effect of RIPC-induced cardioprotection. Activation of STAT5 protein was significantly higher in the Stat5fl/fl+RIPC + I/R group than in the Stat5fl/fl+I/R group, while there was no significant difference between the Stat5-cKO + RIPC + I/R and the Stat5-cKO + I/R group. Further analyses with heart tissues detected decreased protein expressions of cytochrome c (Cyt c) and cleaved Caspase-3 in the Stat5fl/fl+RIPC + I/R mice, along with increased anti-apoptotic molecules, including B-cell lymphoma-extra large (Bcl-xL) and B-cell lymphoma-2 (Bcl-2); such changes were not noted in the Stat5-cKO + RIPC + I/R mice. Additionally, RIPC increased cardiac hypoxia inducible factor-1 (HIF-1α) and interleukin-10 (IL10) protein levels and caused activation of AKT, phosphatidylinositol 3 kinase (PI3K), and vascular endothelial growth factor in the heart of the Stat5fl/fl mice. However, these changes were completely inhibited by the absence of Stat5., Conclusions: These results suggest that RIPC-induced late cardioprotection against myocardial I/R injury is Stat5-dependent and is correlated with the activation of anti-apoptotic signaling and cardiomyocyte-survival signaling.

Purpose: The purpose of this study was to provide the surgeons with effective and reliable guidelines for surgical decision-making by establishing a scoring system for giant cell tumour (GCTSS) based on evidence and expert opinion., Methods: The modified Delphi technique and analytic hierarchy process were used to establish the GCTSS. The GCTSS was defined and classified based on different surgical methods using data from 207 patients collected retrospectively between October 2003 and December 2014. Finally, prospective data of 40 patients between December 2014 and October 2015 were used to analyze concordance between score categorization and experts' consensus on surgical procedure., Results: A novel GCTSS included pathological fracture, cortical bone destruction, tumour size, and articular surface involved. The total scores ranged from 1 to 12 points. The strategy for each patient was decided: a total score of 1-4 suggested intralesional curettage alone for excellent post-operative function; 5-9 points indicated intralesional curettage with internal fixation for less surgery-related complications; and 10-12 points indicated prosthesis replacement for long-term local control. The κ-statistic for the predictive validity of total score was 0.611. The κ coefficient of each group represented moderate or substantial agreement, which was acceptable. The intraclass correlation coefficient for inter- and intra-observer reliability of total score was 0.831 and 0.740, respectively., Conclusions: The novel GCTSS is a comprehensive scoring system with content validity that can aid surgeons in assessing the aggressiveness or severity of giant cell tumour and might become a prognostic tool for surgical decision-making.

Objective: To observe the effect of combined intervention of electroacupuncture (EA) and astragaloside IV(ASIV) on cardiac hypertrophy and transforming growth factor β 1 (TGF-β 1)/Smad signaling in isoproterenol (ISO) induced cardiac hypertrophy rats, so as to investigate its underlying mechanisms in improving myocardial fibrosis., Methods: A total of 50 SD rats were randomly divided into 5 groups: normal control, model (ISO), Propranolol (PRO)，ASIV and EA＋ASIV groups ( n ＝10 in each group). The myocardial fibrosis model was established by intraperitoneal injection (i.p.) of ISO (10 mg·kg －1 ·d －1 ), once daily for 30 days. Rats of the control group were given normal saline (i.p.), those of the PRO group given with PRO (40 mg·kg －1 ·d －1 , gavage), and those of the ASIV and EA＋ASIV groups were treated by gavage of ASIV (40 mg·kg －1 ·d －1 ), once daily for 30 days. EA (20 Hz, 6 V) was applied to bilateral "Neiguan" (PC 6) for 10 min, once every day for 30 d. The heart mass index (HMI, whole heart weight/body weight) and left ventricular (LV) mass index (LVMI, weight of the LV/body weight) were calculated to assess the state of cardiac hypertrophy. The enzyme linked immunosorbent assay (ELISA) was used to determine the levels of procollagen I carboxy-terminal propeptide (PICP，a marker of extracellular matrix remodeling) and carboxyterminal telopeptide of type I collagen (ICTP, a metabolite of type I collagen) in serum, and Western blot was used to test protein contents of TGF- β 1, Smad 2 / 3, Smad 4, Smad 7 in the left ventricle tissue of the heart., Results: After modeling, the HMI and LVMI, serum PICP and ICTP contents and the expression levels of myocardial TGF-β 1, Smad 2/3 and Smad 4 proteins were significantly increased in the model (ISO) group ( P <0.05), suggesting a deposition of collagen and cardiac hypertrophy, and were considerably decreased in PRO, ASIV and EA＋ASIV groups after the intervention ( P <0.05). The expression level of myocardial Smad 7 protein was significantly lower in the model group than in the normal control group ( P <0.05), and significantly up-regulated in PRO, ASIV and EA＋ASIV groups ( P <0.05). Sirius Red staining of the left ventricular myocardium showed a dense deposition of collagen and a severer myocardial fibrosis in the model group, and a relatively lighter fibrosis in the PRO, ASIV and EA＋ASIV groups. The therapeutic effects of EA＋ASIV were comparable to those of PRO, and were significantly superior to those of ASIV in down-regulating HMI, serum ICTP, and myocardial Smad 2/3 and Smad 4 expression and up-regulating Smad 7 protein ( P <0.05). There were no significant differences among the PRO, ASIV and EA＋ASIV groups in LVMI, PICP and TGF-β 1 levels, and between the PRO and EA＋ ASIV groups in HMI, ICTP, Smad 2/3, Smad 4 and Smad 7 levels ( P > 0.05)., Conclusions: EA stimulation of PC 6 combined with ASIV can relieve cardiac hypertrophy and myocardial fibrosis in rats, which may be associated with its effects in regulating myocardial TGF-β 1/Smad signaling pathway.

Background: This study investigated the clinical implication of FAT2 in the progression, metastasis, and prognosis of gastric cancer., Methods: The expression of FAT2 in 436 clinicopathologically characterized gastric cancer cases and 92 control human non-tumor mucosa were analyzed by immunohistochemistry. Consequently, survival analysis was conducted to investigate the association of FAT2 expression and the development of gastric cancers., Results: FAT2 protein was found highly expressed in 90 of 92 (97.83%) control human non-tumor mucosa, while was highly expressed in 126 of 436 (28.90%) tumors samples and low in 310 of 436 (72.10%). The expression of FAT2 was associated with age, tumor size, depth of invasion, Lauren's classification, lymph node and distant metastases, regional lymph node stage, TNM stage, and prognosis. In particular, for stage I, II, and III tumors patients the 5-year survival rate was lower in those with high expression of FAT2 than those with low expression. In stage IV tumors, the expression of FAT2 was not associated with the 5-year survival rate. Lauren's classification and distant metastases, TNM stage, and expression of FAT2 were independent prognostic factors in the patients with gastric cancer, as revealed by Cox regression analysis., Conclusion: The expression of FAT2 in gastric cancer was significantly associated with lymph node and distant metastases, and poor prognosis. FAT2 was also associated with the collective invasion and influenced the prognosis of those patients., Competing Interests: None., (IJCEP Copyright © 2017.)

Background: Garzê and Aba form the second largest Tibetan-inhabited area of China. Blood services have never been reported for this region before., Objective: To assess the current situation and analyse whether a safe and adequate blood supply has been developed in both Garzê and Aba., Methods: We conducted a longitudinal survey covering the period 2011-2016. The subjects of interest were recruited from non-remunerated voluntary donation, blood testing, clinical transfusion practices and infrastructure of local blood service systems., Results: The donation rate and blood collection volume were below the average levels of both the Sichuan Province and mainland China. Component therapy was widely used, but inappropriate usage of whole blood existed. A lack of national specific standards for people on the plateaus led to local blood transfusions being conducted without full clinical assessment. Endemic and frequently occurring disease, such as hydatid disease and gastrointestinal disease, were inevitable risks for blood utilisation and safety. The potential influence of religious belief and traditions, like 'male-leaving marriages', of Tibetans on donor recruitment and blood safety requires further research., Conclusions: A relatively safe and complete blood service system has been developed in this region. However, there is still an urgent need for comprehensive and effective support from the government in terms of policies and finance. As an epidemic area of hydatid disease and sexually transmitted disease, this region needs to emphasise public health measures, such as blood safety and inappropriate usage of blood products., (© 2017 British Blood Transfusion Society.)

Background: Intralesional excision with curettage is the standard method of giant cell tumor (GCT) treatment, but the ideal filling material after curettage remains controversial. The purpose of this study was to compare the oncological and functional outcomes which underwent cementation or bone grafting after GCT curettage around the knee., Methods: We reported 136 cases with GCTs in distal femur or proximal tibia who accepted curettage from five clinical centers during the last 15 years. All patients were divided into two groups according to filling materials. Recurrence-free survival proportions were used to evaluate oncological outcomes while the Musculoskeletal Tumor Society (MSTS) 93 functional score was used to evaluate functional outcomes. Other parameters including surgical complication, general condition, and radiological classification had been analyzed. The valid statisitical data was analyzed using SPSS 13.0 software., Results: After GCT curettage, 86 patients (63.2%) accepted bone grafting while 50 patients (36.8%) accepted cementation. There was no statistical difference in age, gender, tumor location, radiological classification, fixation, follow-up time, and MSTS 93 functional score between cementation group and bone grafting group. The recurrence-free survival proportions showed that the recurrence rate in bone grafting group was higher than it in cementation group (P = 0.034). Surgical complication was lower in cementation group than that in bone grafting group but without statistically significant difference (P = 0.141)., Conclusions: Parameters including patients' age, gender, tumor location, and radiological classification did not affect surgeons' treatments in cavity filling after GCT curettage. Cementation should be recommended because of easy usage, the similar postoperative knee function with bone grafting, and the better local tumor control than bone grafting.

The bacterial acid-resistant chaperone HdeA is a "conditionally disordered" protein that functions at low pH when it undergoes a transition from a well-folded dimer to an unfolded monomer. The dimer dissociation and unfolding processes result in exposure of hydrophobic surfaces that allows binding to a broad range of client proteins. To fully elucidate the chaperone mechanism of HdeA, it is crucial to understand how the activated HdeA interacts with its native substrates during acid stress. Herein, we present a nuclear magnetic resonance study of the pH-dependent HdeA-substrate interactions. Our results show that the activation of HdeA is not only induced by acidification but also regulated by the presence of unfolded substrates. The variable extent of unfolding of substrates differentially regulates the HdeA-substrate interaction, and the binding further affects the HdeA conformation. Finally, we show that HdeA binds its substrates heterogeneously, and the "amphiphilic" model for HdeA-substrate interaction is discussed.

Although intranasal delivery bypasses the blood-brain barrier (BBB), the anatomical location of the olfactory mucosa and respiratory airflow interference lead to less brain-targeted drug delivery. In addition to intranasal delivery, evidence indicates that facial intradermal injection might be a novel strategy for bypassing the BBB via the trigeminal nerve (TN). The hypothesis was verified by pharmacokinetic evaluation, nasal injury, lymphatic vessels inhibition and immunohistochemistry. Intradermal injection into the rat mystacial pad (i.d.) elevated the brain sub-areas and trigeminal Evans Blue (EB) concentrations, C max and AUC (0-t) . I.d. also increased them in brain sub-areas beyond those of intranasal (i.n.) and intravenous injection (i.v.), especially the pons varolii and the medulla oblongata (sub-areas associated with TN). I.d. injection increased the brain drug targeting efficiency, brain direct transport percentage and brain bioavailability of EB while i.n. injection altered them slightly. Trigeminal transection and nasal injury reduced trigeminal EB with i.d. administration. Trigeminal perineurium, epineurium, perivascular spaces, neurons and Schwann cells were involved in the EB brain-targeted delivery. The lymphatic system mediated EB diffusion from the mystacial pad to the nasal mucosa and the brain. Thus, facial intradermal injection might be a promising strategy for brain-targeting delivery, bypassing the BBB via the trigeminal substructures., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The aims of the present study were to determine the effects of heparin-derived oligosaccharides (HDOs) on vascular intimal hyperplasia (IH) in balloon-injured carotid artery and to elucidate the underlying mechanisms of action. An animal model was established by rubbing the endothelia within the common carotid artery (CCA) in male rabbits. The rabbits were fed a high-cholesterol diet. Arterial IH was determined by histopathological changes to the CCA. Serum lipids were detected using an automated biochemical analysis. Expressions of mRNAs for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1), scavenger receptor class B type I (SR-BI), and ATP-binding cassette transporter A1 (ABCA-1) were analyzed using reverse transcription polymerase chain reaction assays. Expressions of VEGF, VCAM-1, MCP-1, SR-BI and ABCA-1 proteins were analyzed by Western blotting. Enzyme-linked immunosorbent assays were used to quantify expression levels of VEGF and bFGF. Our results showed that administration of HDO significantly inhibited CCA histopathology and restenosis induced by balloon injury. The treatment with HDOs significantly decreased the mRNA and protein expression levels of VEGF, bFGF, VCAM-1, MCP-1, and SR-BI in the arterial wall; however, ABCA-1 expression level was elevated. HDO treatment led to a reduction in serum lipids (total cholesterol, triglycerides, high-density and low-density lipoproteins). Our results from the rabbit model indicated that HDOs could ameliorate IH and underlying mechanism might involve VEGF, bFGF, VCAM-1, MCP-1, SR-BI, and ABCA-1., (Copyright © 2017 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Giant cell tumor (GCT) is a benign, locally aggressive tumor that rarely occurs in the spine. They usually occur in patients between 20 and 40 years of age; some patients with GCT present in hospital with pregnancy. The management of these patients can be challenging. The current study reports a case of GCT located in the sixth thoracic vertebra of a 31-year-old female at 34 weeks of gestation. An osteolytic lesion, 3.5 cm × 3.5 cm in diameter, was identified on CT. An MRI test of the lesion revealed a soft tissue mass involving the T 6-7 vertebrae, extending to the right pedicle of the T 6 vertebra and causing significant spinal cord compression. The patient delivered a healthy boy by cesarean at 35 weeks of gestation. One week later, she underwent a T 6 -T 7 laminectomy, posterior spinal decompression, and instrument fusion. Ultimately, the patient was discharged to inpatient rehabilitation with improved lower extremity strength and returned bowel and bladder function. At the 2-year follow-up, there is no evidence of local reoccurrence and the patient has no constitutional symptoms. The present study also reviews eight similar previously reported cases of GCT in the mobile spine, with a discussion of the demographic characteristics, treatment, and prognosis of pregnant patients. This report adds to the existing literature, making the reader aware that clinical symptoms of GCT in the mobile spine can be misinterpreted as a symptom of pregnancy. For spinal GCT in pregnant women, continued pregnancy and definitive surgery after the baby is born are recommended without any serious nerve damage., (© 2017 Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

A modular and adaptive mass spectrometry (MS)-based platform was developed to provide fast, robust and sensitive host cell protein (HCP) analytics to support process development. This platform relies on one-dimensional ultra-high performance liquid chromatography (1D UHPLC) combined with several different MS data acquisition strategies to meet the needs of purification process development. The workflow was designed to allow HCP composition and quantitation for up to 20 samples per day, a throughput considered essential for real time bioprocess development support. With data-dependent acquisition (DDA), the 1D UHPLC-MS/MS method had excellent speed and demonstrated robustness in detecting unknown HCPs at ≥ 50 ng/mg (ppm) level. Combining 1D UHPLC with sequential window acquisition of all theoretical spectra (SWATH) MS enabled simultaneous detection and quantitation of all HCPs in single-digit ng/mg range within 1 hour, demonstrating for the first time the benefit of SWATH MS as a technique for HCP analysis. As another alternative, a targeted MS approach can be used to track the clearance of specific known HCP under various process conditions. This study highlights the importance of designing a robust LC-MS/MS workflow that not only allows HCP discovery, but also affords greatly improved process knowledge and capability in HCP removal. As an orthogonal and complementary detection approach to traditional HCP analysis by enzyme-linked immunosorbent assay, the reported LC-MS/MS workflow supports the development of bioprocesses with optimal HCP clearance and the production of safe and high quality therapeutic biopharmaceuticals.

Objective: To investigate the clinical feasibility and validity of interspinous fastener (ISF) for lumbar degenerative diseases., Methods: From October 2013 to March 2014, a total of 46 patients suffering from lumbar degenerative diseases underwent posterior lumbar interbody fusion (PLIF) randomly augmented by ISF or pedicle screws. The clinical outcome was primarily measured by Oswestry Disability Index (ODI) score. The minimal clinical important difference (MCID) was defined as an eight-point decrease in ODI. The second clinical outcome measurement was Japanese Orthopedic Association (JOA) score. Interbody fusion rates were evaluated by lumbar plain radiograph and computed tomography (CT) scan. Complications were also compared between groups. Statistical analyses were performed by SPSS version 13.0. Sample size calculation was performed before the study. The type I error α was set at 0.05 and the type II error β at 0.1. Based on these assumptions and adding 10% for possible drop-outs, sample size calculations indicated that a total of 46 patients were required for the study. Parametric data was compared by independent t-test and categorical variables were compared using χ 2 -tests or Fisher exact tests depending on the sample size. A P-value of less than 0.05 was considered significantly statistically different. Fleiss kappa coefficients were calculated for intra-observer and inter-observer reliability., Results: A total of 43 patients completed the follow-up, with 22 cases in the ISF group and 21 patients in the pedicle screws group, respectively. Less intraoperative blood loss and shorter operation time were observed in the ISF group. The mean ODI significantly declined in both groups, with the ISF group's decreasing from preoperative 43.3 ± 8.2 to 21.4 ± 3.5 at 24-month follow-up and the pedicle screws group's decreasing from preoperative 42.9 ± 7.9 to 22.5 ±3.8 at 24-month follow-up, respectively. The ODI changes between groups had no statistical difference (P > 0.05). Of the 43 patients, 33 patients achieved an MCID. The bone fusion rate was 77.3% according to X-rays and 68.2% according to CT scans in the ISF group, and 81.0% according to X-rays and 76.2% according to CT scans in the pedicle screws group at the final follow-up. The intra-observer and inter-observer reliability assessed by the kappa value were 0.93 and 0.89, respectively. One patient in the pedicle screws group demonstrated screw loosening at the 6-month follow-up but was asymptomatic. One patient with spondylolisthesis in the ISF group demonstrated cage subsidence during the follow-up but also without related symptoms., Conclusion: The less invasive ISF combined with PLIF provided comparable clinical outcome and a similar bone fusion rate to pedicle screws. The ISF could potentially serve as a new alternative for lumbar degenerative diseases., (© 2017 Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

Background: In acupuncture practice, the most important step is to confirm the location of a sensitized acupoint which reflects a diagnosis and can be stimulated with a specialized needle to treat the disease. Abnormal symptoms such as hyperalgesia or allodynia at the sensitized acupoints in patients with visceral disorders are considered to be in relation with referred pain and neurogenic inflammation. Yet, limited study has investigated the cutaneous neurochemical changes of the sensitized acuponits., Methods: The resent study developed an animal model of gastric mucosal injury (GMI) by HCl administered into the stomach of the rats. Evans Blue (EB) dye was applied by injection of tail vein after mucosal damage to observe the neurogenic plasma extravasation dots in the skin of the rats. The EB dots extravagated in the skin were compared with locations of acupoints. Immnohistochemistry analysis was used to detect the expression of calcitonin gene-related peptide (CGRP)- or substance P (SP)-labeled nerve fibers, histamine (HA)-, serotonin (5-HT)-, and tryptase-labeled cells in EB dots. Images were recorded and analyzed by Confocal imaging system and Olympus Image Processing Software., Results: The results showed that GMI resulted in neurogenic plasma extravasation in the skin of the acupoints over the back and abdomen, which mostly occurred in the T9-11 dermatomere. The EB extravasation dots appeared after GMI and disappeared gradually during the natural self-recovery of the gastric mucosa. More SP and CGRP positive nerve fibers were distributed in EB dots than that in regions beside EB dots and in the control, mostly distributed in the nerve fibers around both the vessels and root of hair follicle. Mast cells also aggregated and degranulated to release algogenic substances of 5-HT and HA around the vessels in areas of the EB dots., Conclusions: Our results indicates that the mechanism of EB extravasation in the skin of the acupoints induced by GMI are closely related to neurogenic inflammation, and that the high expression of local allergic substances and nociceptive neuropeptides in the local skin including SP, CGRP, HA, 5-HT, and mast cell tryptase may be the underlying mechanism of the acupoint sensitization.

Detection of nanoscale objects is highly desirable in various fields such as early-stage disease diagnosis, environmental monitoring and homeland security. Optical microcavity sensors are renowned for ultrahigh sensitivities due to strongly enhanced light-matter interaction. This review focuses on single nanoparticle detection using optical whispering gallery microcavities and photonic crystal microcavities, both of which have been developing rapidly over the past few years. The reactive and dissipative sensing methods, characterized by light-analyte interactions, are explained explicitly. The sensitivity and the detection limit are essentially determined by the cavity properties, and are limited by the various noise sources in the measurements. On the one hand, recent advances include significant sensitivity enhancement using techniques to construct novel microcavity structures with reduced mode volumes, to localize the mode field, or to introduce optical gain. On the other hand, researchers attempt to lower the detection limit by improving the spectral resolution, which can be implemented by suppressing the experimental noises. We also review the methods of achieving a better temporal resolution by employing mode locking techniques or cavity ring up spectroscopy. In conclusion, outlooks on the possible ways to implement microcavity-based sensing devices and potential applications are provided., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A wide resection of the tumor with or without chemotherapy is the treatment of choice for periosteal osteosarcoma (PO) of the extremities, however, post-operative limb function and quality of life could be compromised. The present study reports two cases of 14-year-old boys who presented with progressively enlarging masses in their right knee regions. Computed tomography and magnetic resonance imaging scans indicated a fusiform space-occupying mass encircling the bone cortex, with stippled calcification. A diagnosis of PO was suspected. The histological findings confirmed the diagnosis of intermediate PO. Pre-operative chemotherapies were started, and good responses were detected by clinical evaluation and histological findings. Surgeries preserving the functional structures, including neurovascular bundles, tendons, muscles and epiphyses, were performed, followed by routine chemotherapy. The two patients experienced disease-free survival with follow-up times of 37 and 108 months, respectively. The patients were satisfied with the results of the treatment and they returned to normal life activities. These two cases indicated that a marginal resection of the tumor in conjunction with effective neoadjuvant chemotherapy may be an ideal alternative treatment for intermediate PO, since survival along with well-preserved limb function could be guaranteed. By contrast, a wide excision could result in the loss of limb function.

Measurement of free thiols in antibody therapeutics is important for product development and assessment of critical quality attributes. Earlier studies demonstrated fast separation of free thiol variants of IgG1 using reversed-phase high performance liquid chromatography (RP-HPLC) with diphenyl resin. Here, we report using N-tert-butylmaleimide (NtBM) alkylation followed by RP-HPLC and online mass spectrometry for rapid total and domain-specific free thiol characterization of IgG1. By increasing hydrophobicity, NtBM alkylation improves separation of free thiol variants from disulfide-linked main peak species. The unique mass shift by NtBM alkylation offers unambiguous characterization of free thiol variants by online mass spectrometry. Variant peaks separated by RP-HPLC were antibody molecules containing two NtBM-alkylated cysteines, corresponding to IgG1 containing two free thiols before alkylation. Further characterization of the collected fractions of variants by peptide mapping revealed that each variant contained unpaired cysteines located in specific IgG1 domains (CH1, CH3, VH and VL domains). Total molecular-level and domain-specific free thiol content measured by this method correlate well with orthogonal differential alkylation peptide mapping analysis, which measures free thiol level at individual cysteine residues. This method provides high throughput quantitation of total and domain-specific free thiol content in IgG1 molecules, facilitating rapid, comprehensive product and manufacturing process characterization., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The aim of this work is to investigate the effect of electroacupuncture (EA) at PC6 on the hypertension and myocardial hypertrophy in spontaneously hypertensive rats (SHRs). Thirty SHRs were randomized into model, SHR + EA, and SHR + Sham EA group with WKY rats as normal control. EA was applied once a day in 8 consecutive weeks. The blood pressure (BP), cardiac function, and hypertrophy as well as the underlying mechanisms were investigated. After EA treatment, the enhanced BP in SHR + EA group was significantly lower compared to both the period before EA and model group. Echocardiographic, morphological studies showed that the enhanced left ventricular anterior and posterior wall end-diastolic (LVAWd and LVPWd) thickness, diameters and cross-sectional area (CSA) of cardiac myocyte, as well as the ratio of heart weight to body weight (HW/BW), were markedly diminished in SHR + EA group, while the reduced left ventricular ejection fraction, left ventricular short axis fraction shortening, and E/A ratio were significantly ameliorated. The levels of Angiotensin-converting enzyme (ACE) and Angiotensin II Type 1 and 2 receptors (AT1R, AT2R) in SHRs were also significantly attenuated by EA. The results suggest that EA at bilateral PC6 could arrest the hypertension development and ameliorate the cardiac hypertrophy and malfunction in SHRs, which might be mediated by the regulation of ACE, AT1R, and AT2R., Competing Interests: The authors declare that there is no conflict of interests regarding the publication of this paper.

Many traumatic brain injury (TBI) survivors sustain neurological disability and cognitive impairments due to the lack of defined therapies to reduce TBI-induced blood-brain barrier (BBB) breakdown. Exogenous basic fibroblast growth factor (bFGF) has been shown to have neuroprotective function in brain injury. The present study therefore investigates the beneficial effects of bFGF on the BBB after TBI and the underlying mechanisms. In this study, we demonstrate that bFGF reduces neurofunctional deficits and preserves BBB integrity in a mouse model of TBI. bFGF suppresses RhoA and upregulates tight junction proteins, thereby mitigating BBB breakdown. In vitro, bFGF exerts a protective effect on BBB by upregulating tight junction proteins claudin-5, occludin, zonula occludens-1, p120-catenin, and β-catenin under oxygen glucose deprivation/reoxygenation (OGD) in human brain microvascular endothelial cells (HBMECs). Both the in vivo and in vitro effects are related to the activation of the downstream signaling pathway, PI3K/Akt/Rac-1. Inhibition of the PI3K/Akt or Rac-1 by specific inhibitors LY294002 or si-Rac-1, respectively, partially reduces the protective effect of bFGF on BBB integrity. Overall, our results indicate that the protective role of bFGF on BBB involves the regulation of tight junction proteins and RhoA in the TBI model and OGD-induced HBMECs injury, and that activation of the PI3K/Akt /Rac-1 signaling pathway underlies these effects.

The blood-spinal cord barrier (BSCB) plays important roles in the recovery of spinal cord injury (SCI), and caveolin-1 is essential for the integrity and permeability of barriers. Basic fibroblast growth factor (bFGF) is an important neuroprotective protein and contributes to the survival of neuronal cells. This study was designed to investigate whether bFGF is beneficial for the maintenance of junction proteins and the integrity of the BSCB to identify the relations with caveolin-1 regulation. We examined the integrity of the BSCB with Evans blue dye and fluorescein isothiocyanate-dextran extravasation, measured the junction proteins and matrix metalloproteinases, and evaluated the locomotor function recovery. Our data indicated that bFGF treatment improved the recovery of BSCB and functional locomotion in contusive SCI model rats, reduced the expression and activation of matrix metalloproteinase-9, and increased the expressions of caveolin-1 and junction proteins, including occludin, claudin-5, p120-catenin, and β-catenin. In the brain, in microvascular endothelial cells, bFGF treatment increased the levels of junction proteins, caveolin-1 small interfering RNA abolished the protective effect of bFGF under oxygen-glucose deprivation conditions, and the expression of fibroblast growth factor receptor 1 and co-localization with caveolin-1 decreased significantly, which could not be reversed by bFGF treatment. These findings provide a novel mechanism underlying the beneficial effects of bFGF on the BSCB and recovery of SCI, especially the regulation of caveolin-1., Competing Interests: The authors declare no conflict of interest.

Cucumber (cv. Zhongnong 26) was used as material, the effects of NO 3 - -N/NH 4 + -N ratios on growth and physiological characteristics of cucumber seedlings under suboptimal temperature and light intensity (18 ℃/10 ℃,180 ± 20 μmol·m -2 ·s -1 ) were studied. Total nitrogen in the nutrient solution was equal and three NO 3 - -N/NH 4 + -N ratios, 26:2, 21:7 and 14:14, were applied as treatments. The results showed that cucumber treated by NO 3 - -N/NH 4 + -N=21:7 had the longest total root length, the biggest root volume and root surface area, and the maximum number of root tips. H + -ATPase activity and relative expression of genes encoding nitrate transporter (NRT) and ammonium transporter (AMT) in cucumber roots were increased significantly by the treatment of NO 3 - -N/NH 4 + -N=21:7. In addition, nitrate reductase (NR), glutamine synthetase (GS) and glutamate synthase (GOGAT) in cucumber leaves under the treatment of NO 3 - -N/NH 4 + -N=21:7 were higher. As a result, the nitrogen content and biomass of cucumber were significantly increased. Compared with the plants under the treatment of NO 3 - -N/NH 4 + -N=26:2 or 14:14, cucumber seedlings under the treatment of NO 3 - -N/NH 4 + -N=21:7 had the highest biomass and total dry mass (DM) which were increased by 14.0% and 19.3% respectively under suboptimal temperature and light intensity. In conclusion, under suboptimal environmental conditions, NO 3 - -N/NH 4 + -N ratio could be adjusted to increase nitrogen absorption and metabolism of cucumber and alleviate the de-trimental effects caused by suboptimal conditions and promoted the cucumber growth.

Major wound complications of the extremities, following wide tumor resection and reconstruction for soft-tissue sarcomas (STSs), remain a challenge for limb-sparing surgery. Furthermore, STSs with ulceration or impending ulceration predispose patients to an increased risk of post-operative infection. The present study was conducted to assess the efficacy of negative pressure wound therapy (NPWT) in preventing wound complications associated with surgical treatment of STSs with ulceration or impending ulceration, in patients treated between February 2012 and January 2013. A total of 5 patients, with a mean age of 48 years (range, 24-68 years), were enrolled in the present study. The diagnoses consisted of undifferentiated pleomorphic sarcoma (n=2), leiomyosarcoma (n=1), synovial sarcoma (n=1) and epithelioid sarcoma (n=1). According to American Joint Committee on Cancer criteria, 3 cases were stage III tumors, and the remaining 2 cases were of stages IIA and IIB, respectively. A total of 3 patients exhibited ulceration at diagnosis, and the remaining patients demonstrated impending ulceration. The mean wound area following wide resection of the tumor was 73 cm 2 (range, 45-110 cm 2 ). A continuous suction mode, with pressures measuring -200 to -300 mmHg, was used for 7-10 days on the soft-tissue defects as preparation for wound closure. Soft-tissue reconstruction included muscle flaps (n=2) and skin grafts (n=5). No major wound complications occurred. Post-operative functional and cosmetic outcomes were acceptable. A single patient demonstrated local recurrence 12 months after surgery and re-excision of the tumor was performed. All patients remained alive at the conclusion of follow-up, with a mean follow-up time of 26 months (range, 12-36 months). The present study demonstrated that NPWT is effective and safe when used as an adjunct to wound closure following resection of extremity STS with ulceration/impending ulceration.

The imbalance of autonomic nervous function after myocardial ischemia (MI) accelerates the development of ischemic myocardial injury. The beta-adrenergic receptor (β-AR) and muscarinic acetylcholine receptor (M-AChR) are the main receptor types of the cardiomyocyte and the binding sites of the cardiac sympathetic and parasympathetic transmitters. In the present paper, the authors review recent progress of experimental researches on the mechanisms of acupuncture therapy in the prevention and treatment of MI, mainly focusing on the signaling of β 1 -AR and M 2 -AChR subtypes. Results showed that acupuncture could attenuate the injury of myocardial ischemia via down-regulating the over expression of β 1 -AR and up-regulating the expression of M 2 -AChR protein. Correspondingly, acupuncture intervention could reverse the abnormal expression of the intracellular stimulatory G protein and inhibitory G protein caused by MI, and suppress the activity of cellular adenylate cyclase. In addition, acupuncture could also enhance the activity of nitric oxide synthase and the level of nitric oxide in the ischemic cardiomyocytes, and increase the synthesis of cyclic guanosine monophosphate, ultimately reducing the level of cyclic adenosine 3', 5'monophosphate, and lowering the activity of protein kinase A and the inward flow of Ca 2+ in cardiomyocytes. In this way, the ischemic myocardial injury is improved at last. However, some results of acupuncture intervention are not identical and need being confirmed further by more approaches.

Caloric diet, such as fat and sugar intake, has rewarding effects, and has been indicated to affect the responses to addictive substances in animal experiments. However, the possible association between sucrose reward and the motivation for addictive drugs remains to be elucidated. Thus, we carried out behavioral tests after sucrose self-administration training to determine the effects of sucrose experience on rats' motivation for cocaine, locomotor sensitivity to cocaine, basal locomotor activity, anxiety level, and associative learning ability. The sucrose-experienced (sucrose) group exhibited higher lever press, cocaine infusion and break point, as well as upshift of cocaine dose-response curve in cocaine self-administration test, as compared with the control (chow) group. Additionally, despite similar locomotor activity in open field test and comparable score in cocaine-induced conditioned place preference, the sucrose group showed higher cocaine-induced locomotor sensitivity as compared with the chow group. The anxiety level and the performance in vocal-cue induced fear memory were similar between these two groups in elevated plus maze and fear conditioning tests, respectively. Taken together, our work indicates that sucrose experience promotes the rats' motivation for cocaine.

Stathmin 1 (STMN1) is a biomarker in several types of neoplasms. It plays an important role in cell cycle progression, mitosis, signal transduction and cell migration. In ovaries, STMN1 is predominantly expressed in granulosa cells (GCs). However, little is known about the role of STMN1 in ovary. In this study, we demonstrated that STMN1 is overexpressed in GCs in patients with polycystic ovary syndrome (PCOS). In mouse primary GCs, the overexpression of STMN1 stimulated progesterone production, whereas knockdown of STMN1 decreased progesterone production. We also found that STMN1 positively regulates the expression of Star (steroidogenic acute regulatory protein) and Cyp11a1 (cytochrome P450 family 11 subfamily A member 1). Promoter and ChIP assays indicated that STMN1 increased the transcriptional activity of Star and Cyp11a1 by binding to their promoter regions. The data suggest that STMN1 mediates the progesterone production by modulating the promoter activity of Star and Cyp11a1. Together, our findings provide novel insights into the molecular mechanisms of STMN1 in ovary GC steroidogenesis. A better understanding of this potential interaction between STMN1 and Star in progesterone biosynthesis in GCs will facilitate the discovery of new therapeutic targets in PCOS.

Objectives: To demonstrate the functional results and emotional acceptance after scapulectomy for various malignant shoulder tumors., Methods: Eight patients with malignant shoulder tumors who had undergone scapulectomy between April 2004 and March 2014 were retrospectively reviewed. They comprised seven men and one woman their mean age was 54 years (range, 24-69 years). All patients were diagnosed by pathological examination of biopsy specimens. The tumors were metastatic in four cases, having originated from a primary carcinoma of the liver in one patient, the lung in one patient and the kidney in two patients. The other four patients had primary malignant tumors in their scapulae, specifically, two scapular malignant fibrous histiocytomas, one scapular Ewing sarcoma and one soft tissue synovial sarcoma. The four patients with metastases were staged as III, and other four were staged as IIB. Six patients underwent total and two subtotal scapulectomy. The remaining soft tissues were sutured together directly in seven of the patients. The remaining patient, who had soft tissue synovial sarcoma, required transfer of a pedicle latissimus dorsi muscle flap. The functional results and emotional acceptance were evaluated by clinician using the Musculoskeletal Tumor Society (MSTS) scoring system., Results: The average duration of follow-up was 22.8 months. Four patients were continuously disease-free, three patients developed metastases and died of disease within 12 months of surgery and one patient with a scapular metastasis from the kidney survived with pulmonary nodules. No major complications, including infection or dislocation, occurred during or after surgery. The mean MSTS score was 16.3 (54%), which is similar to that previously reported in other studies of scapulectomy. There were no local tumor recurrences and only one patient developed pulmonary metastases. These outcomes are similar to those reported for scapular prostheses and there were fewer complications than in patients treated with allografts. The mean emotional acceptance score was 3.6 (72.5%)., Conclusions: Performing scapulectomies on patients with malignant shoulder tumors without prostheses or allograft reconstruction achieves good functional results and emotional acceptance with a low rate of complications., (© 2016 Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

In order to explore reasonable utilization of vegetable residue, the effects of application of different levels (0, 15, 20, 30 t·hm -2 ) of tomato and cucumber residue compost on soil pro-perties, the leaf photosynthesis and the yield of vegetables were studied in solar greenhouse. The results showed that both kinds of vegetable compost could reduce soil bulk density, increase soil organic matter content, soil microbial biomass and soil enzyme activity, promote plant growth, and improve vegetable yield and quality. The more amount compost returned, the better was the effect on plant growth and yield. The positive effect of the vegetable residue compost application in the second cucumber growing season was more noticeable than that in the first tomato growing season, which indicated the application of vegetable residue compost could effectively promote soil fertility, increase vegetable yield and improve vegetable nutrient quality.

Objective: To evaluate the result of en bloc resection and reconstruction of the distal radius with a non-vascularized fibular autograft for giant cell tumor (GCT) of bone., Methods: Between 2005 and 2015, 12 eligible patients (seven males, five females, mean age 31.3 years) with grade III GCT of the distal radius were treated by en bloc resection and reconstruction with non-vascularized proximal fibular autografts in four Chinese institutions (members of Giant Cell Tumor Team of China). The patients had a clinical and radiographic review every 6 months for the first 2 years then annually thereafter. The functional, oncologic and radiological outcomes of the patients were analyzed., Results: The mean duration of follow-up was 39.6 months. Bony union was achieved in all cases. None of the patients were dissatisfied with the shape and appearance of the wrist. The mean MSTS score was 25.23 ± 2.38 (range, 22-29). The mean DASH score was 13.0 (range, 6.7-33.3). The average range of motion of the wrist was: 35.8° ± 14.5° of extension, 14.0° ± 8.4° of flexion, 15.5° ± 6.7° of radial deviation, 19.4° ± 10.1° of ulnar deviation, 57.2° ±18.9° of pronation and 44.0° ± 24.8° of supination. The average percentage of grip strength was 55.2% ± 29.0% compared with that of the contralateral side. One localized soft tissue recurrence occurred; it was successfully managed by excision. Lung metastases developed postoperatively in one case and were treated by gamma knife radiotherapy. There was radiographic evidence of radiocarpal arthritis in eleven patients, bone resorption in ten, distal radioulnar joint diastasis in six, ulnar deviation of the wrist in seven, subluxation of the carpal bone in three and dislocation of the carpal bone in one patient., Conclusions: Reconstruction with a non-vascularized proximal fibular autograft is a reasonable option after en bloc resection of the distal radius for giant cell tumor of bone., (© 2016 Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

Objective: To study the impacts of tumor location, nature and extent of bone destruction on selection of operative protocol for extremity osteosarcoma (OS)., Methods: The medical records of 201 patients with extremity OS treated in our institute from December 1999 to June 2014 were retrospectively reviewed. Ninety eligible patients (56 males and 34 females) of average age 20 ± 11 years (range, 4-40 years) were enrolled. Tumor locations were categorized as diaphyseal (4; diaphysis group) or juxta-articular (86); the latter being subclassified as with (51, type III, epiphysis group) or without invasion beyond the epiphyseal line or plate (35, type I and II, metaphysis group) according to MRI images. Tumor nature (osteogenic, 51; osteolytic, 39) was determined radiologically. Extent of bone destruction was quantitated according to Mirel's scoring system to obtain an "invasion score". Regular postoperative follow-up included physical examination and imaging evaluation., Results: Fifty-four patients underwent biological reconstruction and 36 mechanical reconstruction. The mean follow-up duration was 51 months (range, 6-176 months, including four deaths within 12 months). Biological reconstruction was performed more frequently in the diaphysis and metaphysis groups (31/39, 79.5%) than mechanical reconstruction (8/39, 20.5%, P < 0.05). Biological reconstruction and articular preservation were associated with more satisfactory limb function (MSTS scores: 25.0 ± 3.3 and 25.1 ± 3.6) than mechanical reconstruction and articular resection (MSTS scores: 23.4 ± 3.7 and 23.1 ± 3.4, P < 0.05). Reconstruction methods and articular preservation had no relationship with overall or tumor-free survival (P > 0.05). Osteolytic lesions were associated with more extensive bone destruction than osteogenic lesions according to invasion scores (P < 0.05). Following biological reconstruction, high invasion scores (>8) had a 13.5-fold risk of fracture compared with low scores (≤8) (P < 0.05). Twenty-one subjects had recurrences, 30 metastases and 26 died. Postoperative complications included infection (6), fracture (10), and prosthesis loosening (4). Kaplan-Meier analysis indicated 5- and 10-year survival rates of 68.9% and 62.8%, respectively, and 5- and 10-year tumor-free survival rates of 66.7% and 57.8%, respectively., Conclusion: Selection of limb salvage operative protocol for extremity OS should rely on tumor location, nature and extent of bone destruction. Regardless of tumor site, mechanical reconstruction is indicated for tumors with high invasion scores (>8), whereas biological reconstruction is preferred for those with low invasion scores (≤8). Tumors sparing the epiphyseal line or plate are ideal candidates for articular preservation., (© 2016 Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

Giant cell tumor of the tendon sheath (GCTTS), also termed tendosynovial giant cell tumor, is a benign, slow-growing tumor that originates from the tendon sheath or bursa. GCTTS of the foot and ankle is much less frequently reported compared with GCTTS of the hand and knee. However, GCTTS should be considered as a differential diagnosis of soft tissue tumors of the foot and ankle. The optimal treatment strategy for GCTTS in the foot and ankle is controversial due to a scarcity of cases. The present study reports the case of a patient that presented with localized intra-articular GCTTS originating from the capsule of the ankle, which is a rare anatomical location for this tumor. Considering the proximity of the tumor to the adjacent non-tumorous structures, a less radical but complete resection of the tumor was performed, followed by a hydrogen peroxide lavage. There was no evidence of recurrence during a follow-up period of 12 months, and adjuvant radiotherapy was not administered to the patient. A pre-operative diagnosis for GCTTS in the foot and ankle is mainly based on the findings of clinical examination and magnetic resonance imaging, which also facilitates the determination of a surgical strategy. For a localized tumor, an integral resection, as opposed to a radical resection, with a hydrogen peroxide lavage may result in a favorable prognosis. However, the optimal treatment for diffuse GCTTS remains to be identified.

Twenty-two 2-aryl-9-methyl-3,4-dihydro-β-carbolin-2-ium bromides along with four 9-demethylated derivatives were synthesized and characterized by spectroscopic analysis. By using the mycelium growth rate method, the compounds were evaluated for antifungal activities in vitro against six plant pathogenic fungi, and structure-activity relationships (SAR) were derived. Almost all of the compounds showed obvious inhibition activity on each of the fungi at 150 μM. For all of the fungi, 10 of the compounds showed average inhibition rates of >80% at 150 μM, and most of their EC50 values were in the range of 2.0-30.0 μM. SAR analysis showed that the substitution pattern of the N-aryl ring significantly influences the activity; N9-alkylation improves the activity, whereas aromatization of ring-C reduces the activity. It was concluded that the present research provided a series of new 2-aryl-9-alkyl-3,4-dihydro-β-carbolin-2-iums with excellent antifungal potency and structure optimization design for the development of new carboline antifungal agents.

Objective: Matrix metalloproteinase-9 (MMP-9) plays an important role in the remodeling of the extracellular matrix in atherosclerosis plaques. Autophagy protects macrophages against the processes of vascular disease. Our research explores how autophagy plays roles in macrophages to secret MMP-9., Methods and Results: In response to increased doses of oxLDL or CQ we monitored the autophagic flux. Our results revealed that oxLDL was dynamically associated with autophagy and 100 μg/ml oxLDL blocked autophagic flux in THP-1 cells. Moreover p62/SQSTM1 knocking down and CQ respectively inhibited and increased MMP-9 transcriptional expression. These effects were mediated by inhibition of NF-κB., Conclusion: Abundant oxLDL blocked autophagic flux resulting in the aggregation of p62/SQSTM1. Then p62/SQSTM1 was involved in gene expression of MMP-9 via NF-κB-dependent signaling, and thus featuring novel plaque vulnerability properties of the atherosclerotic plaque. Understanding the mechanism that selectively modulates p62/SQSTM1 will provide a novel strategy for anti-atherogenesis., (Copyright © 2016. Published by Elsevier Inc.)

Objective: To investigate the effect of di-(2-ethylhexyl) phthalate (DEHP) exposure on the growth and development of placenta, uterine natural killer (uNK) cell number and angiogenesis at the maternal-fetal interface in pregnant mice., Methods: From day 1 of pregnancy, pregnant mice were exposed daily to DEHP by oral gavage at 125, 250, or 500 mg/kg for 13 consecutive days. The uterine and placental tissues were then harvested for HE staining and immunohistochemistry to examine the effect of DEHP exposure on the growth and development of the placenta and angiogenesis and uNK cell number at the maternal-fetal interface., Results: Compared with the control group, the mice exposed to 500 mg/kg DEHP, but not those exposed to 125 and 250 mg/kg, showed significantly reduced number of embryo implantation (P<0.05). DEHP exposure significantly increased the rate of abortion. DEHP exposure at 125, 250, and 500 mg/kg significantly and dose-dependently lowered the placental weight compared with that in the control group (0.0637±0.0133, 0.0587±0.0176, 0.0524±0.0183 g vs 0.0786±0.0143 g, respectively; P<0.01), and significantly reduced the total area of the placenta and area of spongiotrophoblasts. DEHP exposure resulted in a significant reduction in the number of fetal vascular branches, and collapse and atresia of blood vessels. The mice exposed to DEHP at 125, 250, and 500 mg/kg had significantly lowered numbers of uNK cells (83.2±10.3, 60.7±12.4, and 50.4±14.5/HP, respectively) as compared with the control group (105.1±14.2/HP) at the maternal-fetal interface (P<0.01)., Conclusion: DEHP exposure significantly affects the growth and development of the placenta in mice possibly by suppressing angiogenesis and reducing uNK cell number at the maternal-fetal interface during pregnancy.

Basic fibroblast growth factor (bFGF) may protect stroke patients from cerebral ischemia-reperfusion (I/R) injury. In this study, we report the intranasal use of novel nanoliposomes for the brain delivery of bFGF in a rat model of cerebral I/R. Compared with free bFGF, nanoliposomal therapy was able to significantly improve bFGF accumulation in brain tissues (p<0.05) including the most affected ischemic penumbra regions (e.g. hippocampus, pallium). After intranasal bFGF-nanoliposomal treatment for 3 consecutive days, functional recovery as indicated by improved neurologic deficit score and spontaneous locomotor activity was observed, and the stroke infarct volume was nearly halved (p<0.001) which persisted after 21days. These neuroprotective effects could be blocked by the PI3-K/Akt inhibitor LY294002, indicating the involvement of PI3-K/Akt activation in the therapeutic action. Overall, our results support the intranasal use of nanoliposomal bFGF as an efficient, non-invasive means to bypass the blood-brain barrier for ischemic stroke treatment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Acidic fibroblast growth factor (aFGF) has shown the great potential to prevent the structural and functional injuries caused by diabetic cardiomyopathy (DCM). The present study sought to investigate the preclinical performance and mechanism of the combination therapy of aFGF-nanoparticles (aFGF-NP) and ultrasound-targeted microbubble destruction (UTMD) technique for DCM prevention. From Mason staining and TUNEL staining, aFGF-NP+UTMD group showed significant differences from the diabetes group and other groups treated with aFGF or aFGF-NP. The cardiac collagen volume fraction (CVF) and cardiac myocyte apoptosis index in aFGF-NP+UTMD group reduced to 4.15% and 2.31% respectively, compared with those in the diabetes group (20.5% and 11.3% respectively). Myocardial microvascular density (MCD) in aFGF-NP+UTMD group was up to 35n/hpf, much higher than that in the diabetes group (14n/hpf). The diabetes group showed similar results (MCD, CVF and cardiac myocyte apoptosis index) to other aFGF treatment groups (free aFGF±UTMD or aFGF-NP). Indexes from transthoracic echocardiography and hemodynamic evaluation also proved the same conclusion. These results confirmed that the abnormalities including diastolic dysfunctions, myocardial fibrosis and metabolic could be suppressed by the different extents of twice weekly aFGF treatments for 12 consecutive weeks (free aFGF or aFGF-NP±UTMD), with the strongest improvements observed in the aFGF-NP+UTMD group. Western blot and immunohistochemical analyses of heart tissue samples further revealed the high efficiency of heart-targeted delivery and effective cardioprotection with this combination approach. Overall, this study has generated supportive data that are critical for the translation of a promising DCM prevention strategy., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Context: Theanine, an additive, holds several effects on the central nervous system without toxicity and affects CNS drugs. Theanine bilaterally alters β wave of the EEG with or without caffeine and pentobarbital-induced locomotor activity. Theanine also enhances hypnosis of pentobarbital sodium (PB) and antidepression of midazolam, suggesting there are complicated interactions between theanine and CNS drugs. On the other side, theanine induces glycine release. Glycine potentiates the strychnine toxicity via NMDA receptor activation. Moreover, PB facilitates GABAA receptor activation by GABA, and it is commonly prescribed for strychnine poison. However, what the role that theanine plays in the anticonvulsion of PB against strychnine poison is still unknown., Materials and Methods: Theanine, pentobarbital sodium or strychnine was injected intraperitoneally. EEG was monitored by BIOPAC 16 EEG amplifiers. LD50 of strychnine and hypnotic ED50 of pentobarbital sodium with or without theanine for mice were tested according to Bliss' case., Results: (1) Theanine enhanced the strychnine toxicity. Both theanine and strychnine 1.0 mg/kg increased the power of the β wave. Theanine aggravated that of strychnine 1.0 mg/kg. Theanine attenuated the LD50 of strychnine. (2) Theanine enhanced the anticonvulsion of PB. Theanine increased the power of α, β wave and decreased hypnotic ED50 of PB; PB attenuated strychnine-induced EEG excitation and mortality with or without theanine, and theanine enhanced the effects of PB. Further, theanine enhanced the anticonvulsion of PB dose-dependently against the strychnine toxicity but not the lethal toxicity of strychnine., Discussion and Conclusions: These results indicated theanine interacted with PB and strychnine. Theanine enhanced both the strychnine toxicity and anticonvulsion of PB against strychnine poison.

Objective: To investigate the expression of targeting protein for Xenopus kinesin-like protein 2 (TPX2) in breast cancer tissue and to explore its role in proliferation, migration and invasion of breast cancer cells., Methods: The mRNA and protein expressions of TPX2 in breast cancer tissue and cell lines were assessed by quantitative RT-PCR and Western blot. The effect of TPX2 with RNA interference on proliferation, invasion and migration of breast cancer cells was observed by MTT and Transwell assays., Results: Both mRNA and protein expressions of TPX2 were upregulated in breast cancer tissues compared to tumor-adjacent tissue. TPX2 expression was also upregulated in breast cancer cell lines, and the TPX2 interfered by small interfering RNA could inhibit the proliferation, invasion and migration of breast cancer cells by inhibiting matrix metalloproteinase-2 and matrix metalloproteinase-9., Conclusions: Significantly upregulated TPX2 expression is observed in breast cancer tissue and cells, and contributes to promote the proliferation, migration and invasion of breast cancer cells., (Copyright © 2015 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.)

This paper reports a dramatic reduction in plasmon resonance line width of a single Au nanorod by coupling it to a whispering gallery cavity of a silica microfiber. With fiber diameter below 6 μm, strong coupling between the nanorod and the cavity occurs, leading to evident mode splitting and spectral narrowing. Using a 1.46-μm-diameter microfiber, we obtained single-band 2-nm-line-width plasmon resonance in an Au nanorod around a 655-nm-wavelength, with a quality factor up to 330 and extinction ratio of 30 dB. Compared to an uncoupled Au nanorod, the strongly coupled nanorod offers a 30-fold enhancement in the peak intensity of plasmonic resonant scattering.

Accurate and precise quantitative measurement of product-related variants of a therapeutic antibody is essential for product development and testing. Bispecific antibodies (bsAbs) are Abs composed of two different half antibody arms, each of which recognizes a distinct target, and recently they have attracted substantial therapeutic interest. Because of the increased complexity of its structure and its production process, as compared to a conventional monoclonal antibody, additional product-related variants, including covalent and noncovalent homodimers of half antibodies (hAbs), may be present in the bsAb product. Sufficient separation and reliable quantitation of these bsAb homodimers using liquid chromatography (LC) or capillary electrophoresis-based methods is challenging because these homodimer species and the bsAb often have similar physicochemical properties. Formation of noncovalent homodimers and heterodimers can also occur. In addition, since homodimers share common sequences with their corresponding halves and bsAb, it is not suitable to use peptides as surrogates for their quantitation. To tackle these analytical challenges, we developed a mass spectrometry-based quantitation method. Chip-based nanoflow LC-time-of-flight mass spectrometry coupled with a standard addition approach provided unbiased absolute quantitation of these drug-product-related variants. Two methods for the addition of known levels of standard (multi- or single-addition) were evaluated. Both methods demonstrated accurate and reproducible quantitation of homodimers at the 0.2% (w/w) level, with the single-addition method having the promise of higher analytical throughput.

Taking tomato 'Zhongza 105' as test material, the influences of sodium naphthalene acetate (SNA) on growth and physiological characteristics of tomato seedlings under suboptimal temperature and light condition were investigated. The results showed that the dry mass, vigorous seedling index, root activity, total nitrogen content, net photosynthesis rate (Pn) of tomato seedlings were significantly decreased by suboptimum temperature and light treatment. In addition, the catalase activity and zeatin riboside (ZR) concentration were also reduced. However, the superoxide dismutase, peroxidase activity and the content of abscisic acid (ABA) were increased. Compared with treatment of the same volume distilled water on tomato seedlings under suboptimum temperature and light condition, the dry mass of whole plant and vigorous seedling index of tomato seedlings were significantly increased by 16.4% and 22.9%, as the total N contents in roots and leaves and Pn were also increased by 8.5%, 28.5%and 37.0%, respectively, with the treatment of root application of 10 mg . L-1 SNA. Besides protective enzyme activity and the root activity were improved, the indole acetic acid (IAA) and ZR concentration of tomato were raised, and ABA concentration was reduced. The results indicated that root application of certain concentration of SNA could promote the growth of tomato seedlings by increasing the tomato root activity, protective enzymes activity, Pn and regulating endogenous hormone concentration under suboptimum temperature and light condition.

To understand the diversity in the cell culture harvest (i.e., feedstock) provided for downstream processing, we compared host cell protein (HCP) profiles using three Chinese Hamster Ovary (CHO) cell lines in null runs which did not generate any recombinant product. Despite differences in CHO lineage, upstream process, and culture performance, the cell lines yielded similar cell-specific productivities for immunogenic HCPs. To compare the dynamics of HCP production, we searched for correlations between the time-course profiles of HCP (as measured by multi-analyte ELISA) and those of two intracellular HCP species, phospholipase B-like 2 (PLBL2) and lactate dehydrogenase (LDH). Across the cell lines, proteins in the day 14 supernatants analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) showed different spot patterns. However, subsequent analysis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) indicated otherwise: the total number of peptides and proteins identified were comparable, and 80% of the top 1,000 proteins identified were common to all three lines. Finally, to assess the impact of culture viability on extracellular HCP profiles, we analyzed supernatants from a cell line whose viability dropped after day 10. The amounts of HCP and PLBL2 (quantified by their respective ELISAs) as well as the numbers and major populations of HCPs (identified by LC-MS/MS) were similar across days 10, 14, and 17, during which viabilities declined from ∼80% to <20% and extracellular LDH levels increased several-fold. Our findings indicate that the CHO-derived HCPs in the feedstock for downstream processing may not be as diverse across cell lines and upstream processes, or change as dramatically upon viability decline as originally expected. In addition, our findings show that high density CHO cultures (>10(7) cells/mL)-operated in fed-batch mode and exhibiting high viabilities (>70%) throughout the culture duration-can accumulate a considerable amount of immunogenic HCP (∼1-2 g/L) in the extracellular environment at the time of harvest (day 14). This work also demonstrates the potential of using LC-MS/MS to overcome the limitations associated with ELISA and 2D-PAGE for HCP analysis., (© 2015 Wiley Periodicals, Inc.)

The synthesis, SAR, and in vivo activity of inhibitors of delta-5 desaturase are described. Ring-constraint of the initial series provided access to a variety of in vitro active chemotypes, from which the indazole was selected. Examples from the indazole series displayed in vivo activity in reducing the enzymatic activity of liver delta-5 desaturase., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Long-lasting hypertension results in ventricular remodeling characteristic of myocardial hypertrophy including myocyte hypertrophy and myocardial fibrosis, a pathological process failed to be blocked by the current biomedical means and could eventually lead to heart failure. It has been shown that acupuncture can excite the parasympathetic nerve to counter the overtonic activity of sympathetic nerve, elicit antihypertensive effect and produce cardioprotection via the regulation of calcium signaling-related proteins in myocytes, suggesting its potential role in the management of these disorders. In this paper, the authors review relevant studies and propose, with respect to the autonomic nerve, endocrine system, cytokines and intracellular calcium signaling pathways, several novel ideas concerning the treatment of hypertensive myocardial hypertrophy and cardiac malfunction by acupuncture.

The therapeutic effect of vacuum-assisted closure (VAC) has been confirmed in many types of complex wounds, but there are few relevant reports regarding seawater-immersed wounds. The aim of this study was to determine the effect of VAC on seawater-immersed wound healing under different negative pressures and explore the optimal negative pressure value. Four purebred miniature pigs were used as the experimental animal models. Four acute, symmetrical wounds were made on each side of the spine and designated as the experimental group (wounds with 2 h of seawater immersion) and the control group (wounds without seawater immersion). Wounds were divided into a conventional dressing group and 3 further groups with different VAC therapies (negative pressure at either 120, 180, or 240 mmHg). The extent of wound healing, and speed of granulation growth and re-epithelialization were measured. Bacterial flora distribution in the wounds was observed, and fibronectin levels in the exudate of the wounds were tested. Results showed that seawater immersion aggravated wound injury and that VAC therapy with 180 mmHg negative pressure induced the fastest epidermis migration, obvious edema elimination, significant capillary proliferation, and the highest level of fibronectin, and that in wounds, the proportion of Gram-negative bacteria tended to decrease and that of Gram-positive bacteria tended to increase. Our results show that VAC promotes seawater-immersed wound healing and that 180 mmHg negative pressure may be optimal for wound healing.

There is controversy regarding the impact of infection on long-term prognosis in osteosarcoma patients. Clinical trials and experiments relating to this field could bring reconsideration of immunotherapy for osteosarcoma. The clinical records were reviewed of 125 osteosarcoma patients with a mean follow-up of 5.1±3.9 years (range, 0.5-19.8 years), and a review of the literature was also carried out. Chronic localized infections (but not systemic infection) were determined in 6 patients (4.8%). Similar chemotherapeutic regimens (P=1.00) and histological reactions (P=0.65) were observed in patients with or without infection. Tumor location of proximal tibia (P=0.04) was more common in infected patients. More amputations (P<0.001) were necessitated in infected patients due to uncontrolled infection. The 5-year overall survival rate and event-free survival rate in infected patients were 100%, which were significantly higher than that of the non-infected patients, of whom the rates were 54 and 43% respectively (log-rank test: total survival, P=0.01; tumor-free survival, P=0.01). Distant metastasis was an independent risk factor for survival determined by Cox regression analysis (P<0.001, 95 confidence interval, 1.59-3.98). These findings suggested infection was likely to have positive effects on survival in osteosarcoma patients, however, underlying mechanisms remain to be elucidated. Reconsideration of the association of infection and survival in osteosarcoma patients will help to explore novel therapeutic routes and targets in these patients.

This is a systematic review of articles concerning the morbidity, recurrence rate, treatment and treatment complications of pelvic giant cell tumors (GCTs). The key words "giant cell tumor, pelvis" were used to identify articles which included data on patients with pelvic GCTs in English and Chinese databases of published reports from 1949-2012. The articles were filtered by title, abstract and full text. Thirty-eight articles and 165 patients were identified for this review. Data on all identified patients were studies; data in different articles on the same patients was not used repeatedly. The following patient data were collected where possible and subjected to systematic analysis; age, location of GCT, treatment, follow-up, complications, recurrence and whether alive or dead. The mean age of onset was 33.2 years (range, 14-73 years), the peak ages of onset being between 21 and 40 years. A pronounced sex difference was identified, the male : female ratio being 1:1.7. The acetabulum was the commonest area for pelvic GCTs. Forty-eight tumors were primarily located in the iliac, 60 in the acetabular and 31 in the ischiopubic area. Twenty-seven patients experienced complications of treatment. Patients who had been treated by wide resection had the most complications; these included incisional infection and delayed healing of incisions. Local recurrence was common, having occurred in 39/158 patients (24.6%), comprising 24/72 (33.3%) who had undergone intralesional surgery only; 9/20 (45.0%) who had undergone radiotherapy only; 1/51 (2.0%) who had undergone wide resection; and 5/14 patients (35.7%) who had undergone radiation therapy or cryotherapy plus intralesional surgery. Mortality was low (3.2%, 5/158). Pelvic GCT is not common, the acetabular area appears to the most frequent site and the peak age is the third and fourth decades. Although the recurrence rate is high for all pelvic GCTs, the mortality is low. Treatment has a critical influence on recurrence. In spite of the associated complications, the lower local recurrence rate makes wide resection a reasonable option for patients with extensive and/or aggressive GCTs., (© 2015 Chinese Orthopaedic Association and Wiley Publishing Asia Pty Ltd.)

In this study, we studied the effect of triptolide (TPL) on locomotor function in rats with spinal cord injury. A total of 40 rats were studied after dividing them in two major groups, one was experimental group denoted as TPL group while other was control group denoted as PBS group. Each group was subdivided in four subgroups having five rats each (n = 5). TPL was given intraperitonially at the rate of 5 mg/kg/day in TPL group while PBS was given at the same time interval in the same manner in control group for comparison. A reduction in the cavity area of tissue sections was observed by bright field microscopy from 0.22 ± 0.05 to 0.12 ± 0.05 mm(2) in experimental group after 28 days of treatment while BBB score also improved from 1 to 5 after 14 days of treatment. SPSS software, one way ANOVA, was used for recording statistical analysis and values were expressed as mean ± SEM where P value of <0.01 was considered significant. The expression of I-kBα and NF-kB p65 was also studied using western blotting and after recording optical density (OD) values of western blots. It was observed that treatment with TPL significantly reduced the expression of these factors after 28 days of treatment compared with controls.