Increased vasculogenesis of endothelial cells in hyaluronic acid augmented fibrin-based natural hydrogels - from in vitro to in vivo models.

Vascularisation efficiency plays an essential role in the success of bulk transplantation, while biocompatibility and safety are major concerns in clinical applications. Fibrin-based hydrogels have been exploited as scaffolds for their advantages in biocompatibility, degradability and mass transportation in various forms. However, the mechanical strength and degree of vascularisation remain unsatisfactory for clinical usage. An interpenetrating hydrogel was developed by adding hyaluronic acid (HA) to a fibrin-based natural hydrogel. The vasculogenesis of endothelial cells (human umbilical vein endothelial cells, HUVECs) was characterised within the gel using both in vitro and in vivo animal studies. The in vitro vascular morphology analysis showed 17.9 % longer mean tube length and 14.3 % higher average thickness in 7 d cultivation within the HA-supplemented hydrogel. The in vivo results showed 51.6 % larger total tube area, 1.8 × longer average tube length and 81.6 % higher cell number in the HA-supplemented hydrogel compared to the hydrogel without HA. The experimental results demonstrated better vascularisation and cell recruitment in the HA- supplemented hydrogel. The material properties of the hydrogels were also analysed using atomic force microscopy (AFM). The results revealed 3.7 × higher elasticity of the HA-supplemented hydrogel, which provided better mechanical strength and support for easy handling during procedures. With the demonstrated advantages, the developed hydrogels showed promise for exploitation in various practical clinical applications.

[Imaging characteristics of patients with large vestibular aqueduct syndrome and its relationship with the acoustically evoked short latency negative response].

Objective: To explore the imaging characteristics of large vestibular aqueduct syndrome (LVAS) patients and their relationship with the acoustically evoked short latency negative response (ANSR), so as to provide reference for the diagnosis of LVAS. Methods: Clinical data of 174 patients(334 ears) with LVAS diagnosed and treated by the Department of Otorhinolaryngology Head and Neck Surgery of the First Affiliated Hospital of Guangxi Medical University, from October 2009 to December 2017 were retrospectively analyzed, including 117 males and 57 females, aged from 5 months to 47 years old, with the median age of 4 years and 4 months. ABR and imaging data of patients were collected. Midpoint diameter and the outlet diameter of the vestibular aqueduct were measured on CT images, the midpoint diameter of the intraosseous parts and the extraosseous parts of enlarged endolymphatic sac(EES) were measured on MRI images. The correlation between the above measurements was analyzed by Pearson test using SPSS 17.0. According to whether ASNR was detected in ABR, the above data were divided into two groups, and the differences of the above imaging measurements were compared by the Independent-Sample Test. Results: The average midpoint diameter of the vestibular aqueduct was (1.87±0.58) mm (x±s, the following was the same), and the outlet diameter was (3.07±0.99) mm on CT; the average midpoint diameter of the intraosseous parts in enlarged endolymphatic sac(EES) was (2.39±1.37) mm, and the extraosseous parts was (2.50±2.18) mm on MRI. There was a correlation between the four measurements (P<0.05), among which the midpoint diameter of vestibular aqueduct was strongly positively correlated with the outlet diameter (r=0.760), and the remaining pairs were weakly correlated. ASNR was detected in 241 ears (72.16%,241/334) and undetected in 93 ears (27.84%, 93/334) of the 334 ears with LVAS. Midpoint diameter and the outlet diameter of the vestibular aqueduct in no ASNR group were smaller than the ASNR group, and the difference was statistically significant (t value was 2.814 and 2.754, P<0.05). There was no significant difference in the midpoint diameter of the intraosseous parts and the extraosseous parts of enlarged endolymphatic sac between the two groups, and the difference was no statistically significant(t value was 0.101 and 0.683, P>0.05). Conclusions: There is a strong positive correlation between the midpoint diameter of vestibular aqueduct and the outlet diameter in LVAS patients. There is a certain correlation between the size of vestibular aqueduct and the size of endolymphatic sac. The smaller the diameter of vestibular aqueduct, the lower the occurrence rate of ASNR.

Autoantibody against integrin αv ß3 contributes to thrombocytopenia by blocking the migration and adhesion of megakaryocytes.

Essentials The pathogenesis of immune thrombocytopenia (ITP) has not been fully clarified. We analyzed the role of anti-αvß3 autoantibody in the pathogenesis of ITP in patients. Anti-αvß3 autoantibody impeded megakaryocyte migration and adhesion to the vascular niche. Anti-αv ß3 autoantibody potentially contributes to the pathogenesis of refractory ITP. SUMMARY: Background The pathogenesis of immune thrombocytopenia (ITP) has not been fully clarified. Anti-αvß3 integrin autoantibody is detected in chronic ITP patients, but its contribution to ITP is still unclear. Objectives To clarify the potential role of anti-αvß3 integrin autoantibody in chronic ITP and the related mechanism. Methods Relationship between levels of anti-αvß3 autoantibody and platelets in chronic ITP patients was evaluated. The influence of anti-αvß3 antibody on megakaryocyte (MK) survival, differentiation, migration and adhesion was assessed, and the associated signal pathways were investigated. Platelet recovery and MKs' distribution were observed in an ITP mouse model pretreated with different antibodies. Result In this study, we showed that the anti-αvß3 autoantibody usually coexists with anti-αIIbß3 autoantibody in chronic ITP patients, and patients with both autoantibodies have lower platelets. In in vitro studies, we showed that the anti-αvß3 antibody had no significant effect on the survival and proliferation of MKs, whereas it decreased formations of proplatelet significantly. Anti-αvß3 antibody impeded stromal cell derived facor-1 alpha (SDF-1α)- mediated migration and inhibited the phosphorylation of protein kinase B. Anti-αvß3 antibody significantly inhibited MKs' adhesion to endothelial cells and Fibrogen. The phosphorylation of focal adhesion kinase and proto-oncogene tyrosine-protein kinase Src induced by adhesion was inhibited when MKs were pretreated with anti-αvß3 antibody. In in vivo studies, we showed that injection with anti-αv antibody delayed platelet recovery in a mouse model of ITP. Conclusions These findings demonstrate that the autoantibody against integrin αv ß3 may aggravate thrombocytopenia in ITP patients by impeding MK migration and adhesion to the vascular niche, which provides new insights into the pathogenesis of ITP.

Kinect-based virtual rehabilitation and evaluation system for upper limb disorders: A case study.

OBJECTIVE: To help patients with disabilities of the arm and shoulder recover the accuracy and stability of movements, a novel and simple virtual rehabilitation and evaluation system called the Kine-VRES system was developed using Microsoft Kinect. METHODS: First, several movements and virtual tasks were designed to increase the coordination, control and speed of the arm movements. The movements of the patients were then captured using the Kinect sensor, and kinematics-based interaction and real-time feedback were integrated into the system to enhance the motivation and self-confidence of the patient. Finally, a quantitative evaluation method of upper limb movements was provided using the recorded kinematics during hand-to-hand movement. RESULTS: A preliminary study of this rehabilitation system indicates that the shoulder movements of two participants with ataxia became smoother after three weeks of training (one hour per day). CONCLUSION: This case study demonstrated the effectiveness of the designed system, which could be promising for the rehabilitation of patients with upper limb disorders.

A 15-year-long Southern blotting analysis of FMR1 to detect female carriers and for prenatal diagnosis of fragile X syndrome in Taiwan.

Here, we review the results of Southern blotting analyses of the FMR1 gene performed in our reference laboratory in Taiwan over a 15-year period. In total, 725 high-risk women with a family history of fragile X syndrome (FXS) or idiopathic intellectual disability, 3911 low-risk pregnant women without such family history, and prenatal diagnosis data for 32 foetuses from 24 carrier mothers were included. Only 2 carriers were in the low-risk group, which indicated a prevalence of 1 of 1955 women (95% confidence interval: 1/7156-1/539). A total of 100 carriers were found to be in the high-risk group, thus revealing a significantly higher frequency than the low-risk group (100/725 vs 2/3911, P<0.0001). Eight of the 14 foetuses that inherited the maternal mutant allele were verified to have a full mutation, with the smallest maternal pre-mutation allele carrying 56 CGG repeats. The overall findings confirmed that the carrier prevalence among low-risk women in Taiwan is significantly lower than that reported in western countries. Therefore, the most important step for preventing FXS in Taiwan would be to focus on high-risk women by promoting general awareness of this disease and spreading knowledge regarding the benefits of carrier screening and prenatal testing.

Recurrence of depressive disorders after interferon-induced depression.

Interferon alpha (IFN-α)-treated patients commonly develop depression during the therapy period. Although most IFN-α-induced depressive disorders achieve remission after IFN-α therapy, no studies have examined the long-term mood effects of IFN-α treatment. We conducted a 12-year population-based cohort study of hepatitis C virus (HCV)-infected patients who were older than 20 years and had received IFN-α therapy. The sample was obtained from the Taiwan National Health Insurance Research Database. The cohort included patients with and without IFN-α-induced depression, matched randomly by age, sex and depression history, at a ratio of 1:10. The follow-up started after the last administration of IFN-α and was designed to determine the incidence of recurrent depressive disorder after IFN-α therapy. A total of 156 subjects were identified as having IFN-α-induced depression and achieving full remission after IFN-α therapy. The overall incidence of recurrent depressive disorders among patients with and without IFN-α-induced depression was 56.8 (95% confidence interval (CI), 42.4-76.1) and 4.1 (95% CI, 2.9-5.8) cases, respectively, per 100 000 person-years, P<0.001. The adjusted hazard ratios for recurrent depressive disorder were 13.5 (95% CI, 9.9-18.3) in the IFN-α-treated cohort and 22.2 (95% CI, 11.2-44.2) in the matched cohort for IFN-α-induced depression patients after adjusting for age, sex, income, urbanization and comorbid diseases. IFN-α-induced depression was associated with a high risk of recurrent depression. It was not a transient disease and might be considered an episode of depressive disorder. Continuation therapy might be considered, and further research is needed.

[The effect and factors associated with outcome of extracorporeal membrane oxygenation in refractory cardiogenic shock patients].

Objective: To observe the clinical efficacy and factors associated with outcome of extracorporeal membrane oxygenation (ECMO) in refractory cardiogenic shock patients. Methods: Patients with refractory cardiogenic shock received ECMO treatment in our hospital from May 2013 to November 2015 were retrospectively analyzed. The clinical status before ECMO support, ECMO timing, complications and outcome were observed and analyzed.The hemodynamic data and the amount of vasoactive drugs at 2 hours before ECMO support and at 2, 6, 24 and 48 hours after ECMO support were collected and compared. Results: Ten refractory cardiogenic shock patients were included in this study (5 acute fulminant myocarditis patients, 4 acute myocardial infarction patients, 1 myocardial rupture patient (6 males, 4 females, age ranged 12 to 56 years). Before ECMO, the mean left ventricular ejection fraction (LVEF) was (31.4±10.2)%, the mean score of APACHE Ⅱ was 26.6±10.8. Eight patients developed cardiac arrests and the duration of CPR ranged from 10 to 300 minutes and three patients received IABP. CVP decreased, BP increased, HR decreased, ScVO2 increased, dose of dobutamine decreased at 2 hours after ECMO support. After ECMO support for 6 hours, lactate decreased, dose of norepinephrine decreased. After ECMO support for 24 and 48 hours, hemodynamics became stable and shock was significantly improved. Complication including infection of limb and catheterization site occurred in 3 patients, femoral arterial thrombosis occurred in 2 patients, critical limb ischemia occurred in 2 patients, hemorrhage at the catheterization site occurred in 2 patients. The duration of ECMO ranged from 2 to 220 hours. Nine patients could be weaned off ECMO support and 6 patients survived to hospital discharge. Two patients died due to too late ECMO support, the other two patients died due to severe complication of limb. Conclusions: ECMO can rapidly improve hemodynamic stability of patients with cardiogenic shock. Accurate assessing the timing of ECMO support and decreasing complication of limb play a critical role on improving outcome in refractory cardiogenic shock patients.

p75NTR ectodomain is a physiological neuroprotective molecule against amyloid-beta toxicity in the brain of Alzheimer's disease.

In Alzheimer's disease (AD), neurodegenerative signals such as amyloid-beta (Aß) and the precursors of neurotrophins, outbalance neurotrophic signals, causing synaptic dysfunction and neurodegeneration. The neurotrophin receptor p75 (p75NTR) is a receptor of Aß and mediates Aß-induced neurodegenerative signals. The shedding of its ectodomain from the cell surface is physiologically regulated; however, the function of the diffusible p75NTR ectodomain (p75ECD) after shedding remains largely not known. Here, we show that p75ECD levels in cerebrospinal fluid and in the brains of Alzheimer's patients and amyloid-beta precursor protein (APP)/PS1 transgenic mice were significantly reduced, due to inhibition of the sheddase-tumor necrosis factor-alpha-converting enzyme by Aß. Restoration of p75ECD to the normal level by brain delivery of the gene encoding human p75ECD before or after Aß deposition in the brain of APP/PS1 mice reversed the behavioral deficits and AD-type pathologies, such as Aß deposit, apoptotic events, neuroinflammation, Tau phosphorylation and loss of dendritic spine, neuronal structures and synaptic proteins. Furthermore, p75ECD can also reduce amyloidogenesis by suppressing ß-secretase expression and activities. Our data demonstrate that p75ECD is a physiologically neuroprotective molecule against Aß toxicity and would be a novel therapeutic target and biomarker for AD.

Insilico Approaches in Anesthetic Drug Development: Computer Aided Drug Designing.

OBJECTIVE: Computer Aided Drug DESIGNing is fast becoming an important tool in Drug discovery, and in the field of anesthetic drug development we are the first to use in silico approaches to look for novel anesthetic compounds. DESIGN: The approach of molecular modeling, Virtual screening, Drug-likeness, molecular docking and molecular dynamics simulations (MDS) was employed for this study. RESULT: Our approach of virtual screening Drug-likeness, adsorption, distribution, metabolism, excretion and toxicity analysis of around 50 000 compounds from Inter Bio Screen (IBS) Database have given us top 5 Lead compounds against ASN289 of γ-aminobutyric acid (GABAA) receptor, a common target of known anesthetic compounds. Out of the top 5 Lead compounds one (Lead 5) was selected for further MDS analysis based on its Binding free energy and number of physical interactions with GABAA. CONCLUSION: The MDS analysis of Lead 5 reveals the complex to be stable and thus suitable for further in vitro and in vivo analysis.

Impaired hematopoiesis and delayed thrombopoietic recovery following sublethal irradiation in SRC­3 knockout mice.

The objective of the present study was to investigate the role of the steroid receptor coactivator-3 (SRC-3) in hematopoiesis of mouse bone marrow (BM) following total body irradiation (TBI). SRC-3-/­ mice and wild-type (WT) mice were exposed to 4.5 Gy γ rays. Immunoblotting analysis revealed that the SRC-3 protein (p160) levels in normal BM-nucleated cells in WT were higher than in SRC-3-/­ mice. Furthermore, peripheral blood cell counts, BM cellularity and colony-forming unit (CFU) assays were performed following irradiation. The results showed that peripheral blood cells were significantly lower in number and recovered less rapidly in irradiated SRC-3-/­ mice as compared with control animals. BM-nucleated cell and CFU counts were significantly decreased in SRC-3-/­ mice on the 7th and 14th day. Of note, the recovery of platelet (PLT) and megakaryocytic lineage were more depressed than the granulocytic and erythroid lineage in SRC-3-/­ mice. In conclusion, the present study demonstrated that the hematopoietic ability in SRC-3 knockout mice is severely impaired following a sublethal dose of irradiation.

Retrospective chart review on exposure to psychotropic medications associated with neuroleptic malignant syndrome.

OBJECTIVE: To investigate the association between neuroleptic malignant syndrome (NMS) and levels of antipsychotic exposure. METHOD: Electronic health record data systematically screened from a large mental health service provider in southeast London provided 67 NMS cases which were individually matched with 254 controls on age, gender, and primary psychiatric diagnosis. Data on psychotropic agents, combinations, dose, and dose change of antipsychotic prescriptions over the preceding 5 (oral agents) or 15 days (depot agents) were extracted and compared between groups using conditional logistic regression models. RESULTS: NMS was associated with higher number of antipsychotic agents used, use of first-generation agents or aripiprazole, use of first-generation agents only or cross-generation agents, and higher mean and maximum daily doses. In further analyses, associations with antipsychotics type remained significant when adjusted for dose, but those with dose were attenuated following adjustment for type. The specific use of haloperidol, aripiprazole, depot flupentixol, and benzodiazepines was independently associated with NMS. Non-white ethnicity was also found to be associated with NMS. CONCLUSION: NMS was primarily associated with type of antipsychotic and polypharmacy rather than overall dose. Variation in risk by ethnicity requires further research.

Identification of a novel HLA-A allele, A*02:357, in a Chinese individual.

Human leukocyte antigen (HLA)-A*02:357 differs from A*02:01:01:01 by a single nucleotide at position 840 from A to T exon 4, leading to amino acid substitution from Arg to Ser.

Identification of a novel HLA-A allele A*33:61 in a Chinese individual.

HLA-A*33:61 has two nucleotides change from A*33:03:01 at positions 559 and 560 in exon 3 where A→C and C→G (codon 163 ACG→CGG).

28-day oral toxicity study of the aqueous extract from spider brake (Pteris multifida Poiret) in rats.

Spider brake (Pteris multifida Poiret) is a very important folk herb and a constituent in most of the traditional herbal beverage formulas in Taiwan; however, little toxicological information is available regarding the safety following repeated exposure. The present study was conducted to evaluate the toxicity of aqueous extract from spider brake (SB) in Sprague-Dawley rats on dietary oral gavage at concentrations of 100, 500, and 1000 mg/kg b.w. day for 28 days. There were no adverse effects on general condition, growth, feed and water consumption, feed conversion efficiency, red blood cell and clotting potential parameters, clinical chemistry values, and organ weights except for neutrophils and lymphocytes being slightly diminished in male and female rats at the highest dose, respectively. Necropsy and histopathology findings revealed no treatment-related changes in any of the organs. The results obtained in this study allowed us to conclude that the SB properly utilized in the traditional oral administration could be devoid of any toxic risk.

Identification of Atractylodes plants in Chinese herbs and formulations by random amplified polymorphic DNA.

AIM: An efficient, precise, and sensitive method for identifying Atractylodes plants has been established and will contribute significantly to quality control and scientific analysis in Chinese traditional medicine. METHODS: Twenty primers were applied for setting up the RAPD (randomly amplified polymorphic DNA) markers of Atractylodes plants, Atractylodes lancea DC (A lancea DC), Atractylodes japonica Koidz (A japonica K), and Atractylodes ovata DC (A ovata DC). The primer OPF03, OPF05, and OPF14 could discriminate them successfully. The results were also able to apply on the Chinese formulations with Atractylodes purchased from local markets. RESULTS: RAPD was used to investigate phylogenetic relationships among and within closely related species. RAPD analysis reflects heritable changes in the nucleotides sequence in both the coding and noncoding regions, because it is conducted directly from the DNA level. This work first conducted RAPD analysis of Atractylodes plants to establish their RAPD makers. CONCLUSION: The RAPD markers could be applied extensively in the Chinese herbal formulations.

Quantitative expression profile of androgen-regulated genes in prostate cancer cells and identification of prostate-specific genes.

Quantitative expression profile of androgen-regulated genes (ARGs) was evaluated in the hormone-responsive prostate cancer cell line LNCaP by serial analysis of gene expression (SAGE). A total of 83,489 SAGE tags representing 23,448 known genes or expressed sequence tags (ESTs) and 1,655 potentially novel sequences have unraveled the transcriptome of LNCaP cells, the most common cell line used in prostate cancer research. Comparison of transcripts between control and R1881-treated LNCaP cells revealed the induction of 136 genes and repression of 215 genes in response to androgen (p < 0.05). Strikingly, a high fraction ( approximately 90%) of ARGs identified in our study has not been described as ARGs previously. A number of prostate-specific transcription factors were among the ARGs identified here. Classification of the ARGs on the basis of biochemical functions revealed that a great majority of ARGs identified in our experimental system appear to be involved in regulation of transcription, splicing, ribosomal biogenesis, mitogenesis, bioenergetics and redox processes. One of the novel aspects of androgen signaling included androgen regulation of genes involved in DNA repair/recombination process. By comparing our LNCaP-C and LNCaP-T SAGE libraries with SAGE tag libraries available at the NCBI-SAGE website, we have identified >200 potential prostate specific/abundant transcripts. The discovery of new prostate-specific genes and ARGs provides a unique opportunity to determine the role of these genes in prostate cell growth, differentiation and tumorigenesis.

Monoclonal antibodies against different epitopes of nonstructural protein sigmaNS of avian reovirus S1133.

Ten monoclonal antibodies (MAbs) were prepared against the nonstructural protein sigmaNS of avian reovirus S1133. Eight of them were selected for two-way competitive binding assay after coupling with horseradish peroxidase. The results allowed the definition of three epitopes, designated A, B, and C. Blocking assay of poly(A)-Sepharose binding activity of sigmaNS with MAbs indicated that MAb recognizing epitope B was able to block poly(A) oligomer binding, suggesting that epitope B is involved in ssRNA binding of sigmaNS. An immuno-dot binding assay was used to analyze the effect of denaturation on antibody recognition of the epitopes. All MAbs bound to protein sigmaNS in its native form. After denaturation by boiling in SDS and 2-mercaptoethanol, the binding of MAbs recognizing epitopes B and C was not affected. The reactivity of MAbs recognizing epitope A was fully abolished by denaturation. These results suggest that the binding of MAbs directed against epitope A is conformation-dependent; however, the recognition by MAbs of epitopes B and C is not conformation-dependent. In addition, the results from the cross-reactivity of MAbs to heterologous avian reovirus strains suggest that the three epitopes are highly conserved among these virus strains.

[Changes in aldosterone binding activity of kidney cytosol after stress in rats and the regulation].

To observe the changes in aldosterone binding activity of kidney cytosols after pathological stress in rats and the regulation, binding capacity (Rt) and apparent dissociation constant (Kd) of aldosterone binding activity of kidney cytosols in normal, low-degree or heavy-degree scalded rats were measured by radioligand binding assay using [3H]aldosterone as the ligand. Changes in Rt and Kd of aldosterone binding activity were observed after injection of anti-rat TNF alpha and IL-1 beta antibodies, alpha-melanocyte-stimulating hormone (alpha-MSH) and KPV peptide (Ac-D-Lys-L-Pro-D-Val). The results indicated that there were two types of aldosterone binding activities in kidney cytosol with different Rt and Kd, and the Rt of heavy-degree scalded rats (Rt1: 22.4 +/- 5.4 fmol/mg pro, Rt2: 196.3 +/- 32.5 fmol/mg pro) was lower than that of the control group (Rt1: 41.6 +/- 7.2 fmol/mg pro, Rt2: 317.6 +/- 70.0 fmol/mg pro) (P < 0.01; P < 0.01); while the Rt of low-degree scalded rats (Rt1: 41.4 +/- 5.0 fmol/mg pro, Rt2: 314.8 +/- 45.7 fmol/mg pro) was not significantly different from that of the control group (P > 0.05; P > 0.05). Injection of anti-rat TNF alpha and IL-1 beta antibodies, alpha-MSH and KPV prevented Rt of aldosterone binding activity from decrease in kidney cytosol of rats with heavy-degree scald. These findings suggest that aldosterone binding activity may be down-regulated in heavy-degree scalded rats, but it may be reversed by injection of anti-rat TNF alpha and IL-1 beta antibodies, alpha-MSH and KPV.

Physician-evaluated and self-reported morbidity for predicting disability.

OBJECTIVES: This study compared the predictive validity of physician-evaluated morbidity and self-reported morbidity on disability among adults. METHODS: Subjects from a large national survey (n = 6913) received a detailed medical examination by a physician and were asked about the presence of 36 health conditions at baseline. Disability measured 10 and 15 years later was regressed on the morbidity measures and covariates with tobit models. RESULTS: Although physician-evaluated morbidity and self-reported morbidity were associated with greater disability, self-reports of chronic nonserious illnesses manifested greater predictive validity. Disability was also higher for obese subjects and those of lower socioeconomic status. CONCLUSIONS: The findings demonstrate the predictive utility of self-reported morbidity measures on functional disability.