Search

BACKGROUND The organ system-based medical curriculum is crucial for students in the MD-PhD program of Xiangya School of Medicine at Central South University in China. The combination of Team Based Learning (TBL) and Problem Based Learning (PBL) forms a new case-based teaching model so that students can cooperatively participate in case discussions using multi-disciplinary knowledge. The aim of this study is to utilize TBL-PBL teaching model in the organ system-based medical curriculum, and assess the feedback on teaching arrangement, effectiveness and the academic performance of students. METHODS This is a single institution, two-year study that includes two cohorts of students who participated in two training paradigms, traditional teaching model and TBL-PBL teaching modal. The MD-PhD students enrolled in 2016 (n=100) were selected for traditional teaching model, students enrolled in 2017 (n=100) were selected for TBL-PBL teaching model, in which the learning contents were remodeled and integrated with simulated cases, taught by an interprofessional team. After completing the curriculmn, all students were surveyed on their feedbacks about group teaching arrangement and teaching effectiveness. Academic performance were also assesses by post-session quizzes, periodical tests and the final exams. RESULTS The survey results revealed that in the TBL-PBL teaching group, the percentage of students who thought that the teaching objectives were challenging reached 94%. Meanwhile, for the traditional teaching group, the percentage of students who thought likewise were 84%. The percentage of students in the TBL-PBL group who thought that the innovative teaching mode rendered helpful in consolidating basic medical knowledge structure (89% vs. 78%), promoted student overall thinking skills (94% vs. 84%), communication skills(93% vs. 82%), teamwork skills (95% vs. 82%) and professional abilities (89% vs. 84%) outweighed the traditional teaching group to a significant degree. The academic performance in the TBL-PBL group was improved evidenced by higher class scores(85.52 vs. 80.34), excellence rate (51% vs. 49%) and qualification rate (100% vs. 92%). CONCLUSION The TBL-PBL teaching model improved academic performance, yielded higher student satisfaction and engagement. This innovative teaching model can be used as an alternative to traditional model for medical courses.

BACKGROUND Energy Metabolism and Internal Environment System (EMIES) is a crucial curriculum in the organ-system integrated medical courses for students in the MD-PhD program of Xiangya School of Medicine at Central South University in China. The combination of Team Based Learning (TBL) and Problem Based Learning (PBL) forms a new case-based teaching model so that students can cooperatively participate in case discussions using multi-disciplinary knowledge. The aim of this study is to utilize TBL-PBL teaching model in EMIES curriculum, and assess the feedback on teaching arrangement, effectiveness and the academic performance of students. METHODS This is a single institution, two-year study that includes two cohorts of students who participated in two training paradigms, traditional teaching model and TBL-PBL teaching modal. The MD-PhD students enrolled in 2016 (n = 100) were selected for traditional teaching model, students enrolled in 2017 (n = 100) were selected for TBL-PBL teaching model, in which the learning contents were remodeled and integrated with simulated cases, taught by an interprofessional team. After completing the curriculmn, all students were surveyed on their feedbacks about group teaching arrangement and teaching effectiveness. Academic performance were also assesses by post-session quizzes, periodical tests and the final exams. RESULTS The survey results revealed that in the TBL-PBL teaching group, the percentage of students who thought that the teaching objectives were challenging reached 94%. Meanwhile, for the traditional teaching group, the percentage of students who thought likewise were 84%. The percentage of students in the TBL-PBL group who thought that the innovative teaching mode rendered helpful in consolidating basic medical knowledge structure (89% vs. 78%), promoted student overall thinking skills (94% vs. 84%), communication skills (93% vs. 82%), teamwork skills (95% vs. 82%) and professional abilities (89% vs. 84%) outweighed the traditional teaching group to a significant degree. The academic performance in the TBL-PBL group was improved evidenced by higher class scores (85.52 vs. 80.34), excellence rate (51% vs. 49%) and qualification rate (100% vs. 92%). CONCLUSION The TBL-PBL teaching model improved academic performance, yielded higher student satisfaction and engagement. This innovative teaching model can be used as an alternative to traditional model for medical courses.

Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), which are pivotal sensors of RNA virus invasions, mediate the transcriptional induction of genes encoding type I interferons (IFNs) and proinflammatory cytokines, successfully establishing host antiviral immune response. A few excellent reviews have elaborated on the structural biology of RLRs and the antiviral mechanisms of RLR activation. In this review, we give a basic understanding of RLR biology and summarize recent findings of how RLR signaling cascade is strictly controlled by host regulatory mechanisms, which include RLR-interacting proteins, post-translational modifications and microRNAs (miRNAs). Furthermore, we pay particular attention to the relationship between RLRs and diseases, especially how RLRs participate in SARS-CoV-2, malaria or bacterial infections, how single-nucleotide polymorphisms (SNPs) or mutations in RLRs and antibodies against RLRs lead to autoinflammatory diseases and autoimmune diseases, and how RLRs are involved in anti-tumor immunity. These findings will provide insights and guidance for antiviral and immunomodulatory therapies targeting RLRs.

Purpose: Previous studies proposed that the multidisciplinary team (MDT) consultation could improve tumor staging accuracy and outcomes of patients with gastric malignancy. However, evidence-based reports remain limited. This study aimed to determine the effectiveness of MDT for tumor staging accuracy and outcomes of patients with resectable gastric cancer, and to explore the potential factors affecting its effectiveness. Methods: This retrospective study enrolled 719 gastric cancer patients who underwent gastrectomy in our hospital. After propensity score matching, 378 patients were selected, including 189 in the non-MDT group and 189 in the MDT group. Data regarding baseline characteristics, staging, treatments, and survival were analyzed. Results: The data showed that the staging accuracy in the MDT group and non-MDT group was comparable (53% vs 61% for T stage, 46.1% vs 35.3% for N stage, and 78.3% vs 78.7% for M stage). The MDT group had a higher proportion of preoperative chemotherapy (39.2% vs 28%, p=0.03) and laparoscopic surgery (82.5% vs 72%, p=0.02) than the non-MDT group. However, the achievement of R0 resection was similar in the two groups (93.7% vs 88.9%, p=0.73). There was no significant difference in the 1-year and 3-year overall survival rates between the two groups. Moreover, we observed poor patient compliance when the MDT recommended further examinations, radiotherapy, or chemotherapy before surgical interventions. Conclusion: MDT consultation has limited effects on improving the staging accuracy and treatment outcomes including survival of patients with resectable gastric cancer. Poor patient compliance may be a factor affecting the effectiveness of MDT consultation.

Myeloid-derived suppressor cells (MDSCs) mobilize and migrate from bone marrow to peripheral tissues or immune organs, which is associated with poor prognosis in sepsis. Intervention of MDSCs might be a potential target for the effective treatment of sepsis. In the present study, we demonstrated that IL-1R1 blockade with either recombinant human IL-1R antagonist Anakinra or IL-1R1 deficiency had a protective effect on the liver injury in septic mice. The possible mechanism was that Anakinra treatment and IL-1R1 knockout inhibited the migration of MDSCs to the liver in sepsis, thus attenuating the immune suppression of MDSCs on effector T cells characterized with the decrease in proportion of CD4

For the first time, increased Dp71 in ischemia-reperfusion injured rat heart were identified, both Dp71 mRNA and protein reached its peak expression 8 h after reperfusion. In H

Tumor growth pattern (TGP) and perineural invasion (PNI) at the invasive margin have been recognized as indicators of tumor invasiveness and prognostic events in colorectal cancer (CRC). This study aims to develop a scoring system incorporating TGP and PNI, and further investigate its prognostic significance for CRC risk stratification. A scoring system, termed tumor-invasion score, was established by summing TGP and PNI scores. The discovery cohort (N = 444) and the validation cohort (N = 339) were used to explore the prognostic significance of the tumor-invasion score. The endpoints of the event were disease-free survival (DFS) and overall survival (OS) which were analyzed by the Cox proportional hazard model. In the discovery cohort, Cox regression analysis showed that DFS and OS were inferior for score 4 group compared with score 1 group (DFS, hazard ratio (HR) 4.44, 95% confidence interval (CI) 2.49–7.92, p

Immune checkpoint inhibitors (ICIs) represent a new paradigm in cancer immunotherapy, but can be largely restricted by the limited presence of CD8

Tendinopathy is a complex, multifaceted tendon disease that is often associated with overuse and causes significant health care costs with its high prevalence. At present, the pathogenesis and effective treatment of tendinopathy cannot be fully elucidated. This study aims to deeply explore and analyze the key genes, functional pathways, and immune infiltration characteristics of the occurrence and development of tendinopathy.MethodsThe gene expression profiles of GSE106292, GSE26051 and GSE167226 were downloaded from GEO database. The WGCNA analysis was performed on GSE106292 data set by the R package in R software, and differential gene analysis was performed on GSE26051 and GSE167226 data sets by combining and screening. The gene enrichment analysis of GO and KEGG and immune cell infiltration analysis were performed. Lasso logistic regression, support vector machine (SVM-REF) and Gaussian mixture model (GMMS) algorithm were used to screen and identify early diagnostic genes.ResultsWe have obtained a total of 171 DEGs through WGCNA analysis and screening of different expressed genes. By GO and KEGG enrichment analysis, it was found that these malregulated genes were related to mTOR, HIF-1, MAPK, NF-κB and VEGF signaling pathways. Immunoinfiltration analysis showed that M1 macrophages, activated mast cells and activated NK cells had infiltration significance. MacroD1 may be an early diagnosis gene, and it was found based on Lasso, SVM-REF and GMMS algorithm.ConclusionsBased on comprehensive bioinformatics analysis, we identified the potential early diagnosis genes MACROD1, key regulatory pathways and immune infiltration characteristics of tendinopathy. These key genes and pathways may be used as biomarkers and molecular therapeutic targets for early tendon injury to guide drugs and basic research.

Tendon injuries are one of the most common musculoskeletal disorders that cause considerable morbidity and significantly compromise the patients' quality of life. The innate limited regenerative capacity of tendon poses a substantial treating challenge for clinicians. MicroRNAs (miRNAs) are a family of small non-coding RNAs that play a vital role in orchestrating many biological processes through post-transcriptional regulation. Increasing evidence reveals that miRNA-based therapeutics may serve as an innovative strategy for the treatment of tendon pathologies. In this review, we briefly present miRNA biogenesis, the role of miRNAs in tendon cell biology and their involvement in tendon injuries, followed by a summary of current miRNA-based approaches in tendon tissue engineering with a special focus on attenuating post-injury fibrosis. Next, we discuss the advantages of miRNA-functionalized scaffolds in achieving sustained and localized miRNA administration to minimize off-target effects, and thus hoping to inspire the development of effective miRNA delivery platforms specifically for tendon tissue engineering. We envision that advancement in miRNA-based therapeutics will herald a new era of tendon tissue engineering and pave a way for clinical translation for the treatments of tendon disorders.

Flexor tendon injury is often associated with suboptimal outcomes and results in substantial digit dysfunction. Stem cells have been isolated from several experimental animals for the growing interest and needs of utilizing cell-based therapies. Recently, turkey has been developed as a new large animal model for flexor tendon research. In the present study, we reported the isolation and characterization of bone marrow-derived mesenchymal stem cells (BMSCs) from 8- to 12-month-old heritage-breed turkeys. The isolated cells demonstrated fibroblast-like morphology, clonogenic capacity, and high proliferation rate. These cells were positive for surface antigens CD90, CD105, and CD44, but were negative for CD45. The multipotency of turkey BMSCs was determined by differentiating cells into osteogenic, adipogenic, chondrogenic, and tenogenic lineages. There was upregulated gene expression of tenogenic markers, including mohawk, tenomodulin, and EGR1 as well as increased collagen synthesis in BMP12 induced cells. The successful isolation and verification of bone marrow-derived MSCs from turkey would provide opportunities of studying cell-based therapies and developing new treatments for tendon injuries using this novel preclinical large animal model. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1419-1428, 2019.

Objective The present study was designed to evaluate the functional outcome of stapled transanal rectal resection (STARR) and to examine the relationship between the population density of the interstitial cells of Cajal (ICC) and the efficacy of the STARR operation in the management of obstructed defecation syndrome (ODS) patients. Methods Full-thickness rectal samples were obtained from 50 ODS patients who underwent STARR. Samples were analysed using ICC immunohistochemistry. Clinical and functional parameters obtained with defecography and anorectal manometry were compared with 20 controls. Results ICCs were significantly decreased in patients in the submucosal (SM), intramuscular (IM) and myenteric (MY) regions when compared with the control group (P

// Wei Dong 1, * , Min Zhang 2, * , Yaxi Zhu 3 , Yuanhan Chen 1 , Xingchen Zhao 1 , Ruizhao Li 1 , Li Zhang 1 , Zhiming Ye 1 and Xingling Liang 1 1 Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China 2 Department of Gastroenterology, The Sixth Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China 3 Department of Pathology, The Sixth Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China * These authors have contributed equally to this work Correspondence to: Xingling Liang, email: xinlingliang_ggh@163.com Keywords: intestinal epithelial cells, inflammatory bowel disease, colitis, necroptosis, necrosulfonamide Received: June 12, 2017 Accepted: September 05, 2017 Published: September 30, 2017 ABSTRACT Objective: This study aimed to investigate the protective effect of the necroptosis inhibitor necrosulfonamide (NSA) on intestinal epithelial cells using a novel in vitro necroptosis model that mimics inflammatory bowel disease (IBD). Methods: 2,4,6-trinitrobenzenesulfonic acid (TNBS) was perfused into the rectum of BALB/c mice to established a colitis model. Pathologic injury and cell death were evaluated. A novel in vitro model of necroptosis was established in Caco-2 cells using TNF- α and Z-VAD-fmk, and the cells were treated with or without NSA. Morphologic changes, manner of cell death and the levels of phosphorylation of receptor-interacting protein kinase 3 (p-RIPK3) and mixed-lineage kinase domain-like (p-MLKL) were detected. Results: In the TNBS-induced colitis in mice, TUNEL-positive and caspase-3-negative cells were observed in the intestinal mucosa, and p-RIPK3 was found to be elevated. Under the stimulation of TNF- α and Z-VAD-fmk, the morphologic damage in the Caco-2 cells was aggravated, the proportion of necrosis was increased, and the level of p-RIPK3 and p-MLKL were increased, confirming that the regulated cell death was necroptosis. NSA reversed the morphological abnormalities and reduced necrotic cell death induced by TNF- α and Z-VAD-fmk. Conclusion: NSA can inhibit necroptosis in intestinal epithelial cells in vitro and might confer a potential protective effect against IBD.

The selective hydrogenation of a molecule containing several unsaturated groups is a very important issue and has attracted much more attention in the chemical synthesis. Herein, a highly selective catalyst of Pt-Bi/TiO 2 was first time developed for the hydrogenation of 3-nitrostyrene (NS) to 3-aminostyrene (AS), with it an excellent selectivity to AS of 96% was achieved at the complete conversion. The influence of Bi on the catalytic performance was well discussed by using X-ray diffraction (XRD), transmission electronic microscopy (TEM), CO pulse chemisorption, hydrogen temperature-programmed reduction (H 2 -TPR), X-ray photoelectron spectroscopy (XPS), diffuse reflectance Fourier transform infrared (DRFTIR) spectroscopy of adsorbed CO, and thermo gravimetric analysis (TGA) and experiment data. With increasing Bi loading, Bi-rich surface was formed, and the terraces sites of Pt disappeared as it transferred to the low coordinated Pt gradually. Thus, high selectivity of AS was obtained over Pt-Bi/TiO 2 for the hydrogenation of AS to 3-ethylaniline (EA) was lowered.

Aims Immunosuppressive myeloid-derived suppressor cells (MDSCs) continuously expand and lead to poor outcome during sepsis. The activation of liver X receptor (LXR) can mitigate sepsis-induced liver and myocardial damage. This study aims to determine whether LXR plays a protective role in sepsis by regulating MDSCs. Main methods Cecal ligation and puncture(CLP)was used to induce sepsis in mice. The mice were then treated with LXR agonist GW3965 (3 mg/kg) or vehicle 1 h, 6 h, 12 h, 24 h, 48 h, 72 h postoperatively. The effect of LXR on the survival rate and multi-organ injury induced by sepsis was evaluated by survival analysis, histological staining, biochemical analysis and ELISAs. The percentages of MDSCs and T cells were detected using flow cytometry. The mRNA expressions of LXR and ATP-binding cassette transporter A1 (ABCA1) were measured using real-time quantitative PCR (RT-qPCR). ABCA1 protein level was determined using immunofluorescence staining. Key findings LXR agonist GW3965 treatment improved the survival of septic mice, accompanied by reduced multi-organ injury and a decreased level of inflammatory cytokines. Furthermore, GW3965 treatment decreased MDSCs abundance in spleen by boosting the apoptosis of spleen MDSCs, therefore ameliorating their immunosuppressive activity. Meanwhile, bacteria clearance in tissues was enhanced after the GW3965 administration in septic mice. Mechanistically, GW3965 activated LXRβ and its downstream target ABCA1 to initiate the apoptosis of spleen MDSCs. Significance These findings provide new insights into the relationship between LXR and MDSCs in sepsis, thus revealing a potentially effective approach to target the immunosuppression of sepsis.

To explore the function of Dof transcription factors during kernel development in maize, we first identified Dof genes in the maize genome. We found that ZmDof3 was exclusively expressed in the endosperm of maize kernel and had the features of a Dof transcription factor. Suppression of ZmDof3 resulted in a defective kernel phenotype with reduced starch content and a partially patchy aleurone layer. The expression levels of starch synthesis-related genes and aleurone differentiation-associated genes were down-regulated in ZmDof3 knockdown kernels, indicating that ZmDof3 plays an important role in maize endosperm development. The maize endosperm, occupying a large proportion of the kernel, plays an important role in seed development and germination. Current knowledge regarding the regulation of endosperm development is limited. Dof proteins, a family of plant-specific transcription factors, play critical roles in diverse biological processes. In this study, an endosperm-specific Dof protein gene, ZmDof3, was identified in maize through genome-wide screening. Suppression of ZmDof3 resulted in a defective kernel phenotype. The endosperm of ZmDof3 knockdown kernels was loosely packed with irregular starch granules observed by electronic microscope. Through genome-wide expression profiling, we found that down-regulated genes were enriched in GO terms related to carbohydrate metabolism. Moreover, ZmDof3 could bind to the Dof core element in the promoter of starch biosynthesis genes Du1 and Su2 in vitro and in vivo. In addition, the aleurone at local position in mature ZmDof3 knockdown kernels varied from one to three layers, which consisted of smaller and irregular cells. Further analyses showed that knockdown of ZmDof3 reduced the expression of Nkd1, which is involved in aleurone cell differentiation, and that ZmDof3 could bind to the Dof core element in the Nkd1 promoter. Our study reveals that ZmDof3 functions in maize endosperm development as a positive regulator in the signaling system controlling starch accumulation and aleurone development.

Aims To investigate the prevalence of adult-onset autoimmune diabetes (ADM) and predisposition to autoimmune diseases by quantifying serum organ-specific autoantibodies in people with phenotype of type 2 diabetes (T2D). Materials and methods We included a nationally representative sample of 46 239 adults aged ≥20 years from 14 provinces, of whom 4671 had diabetes, plus 1000 control subjects with normal glucose tolerance (NGT). Participants were screened centrally for autoantibodies to glutamic acid decarboxylase (GAD), islet antigen 2 (IA2) and zinc transporter isoform-8 (Znt8) and were defined as having ADM where positive for these antibodies. We then assayed thyroid peroxidase (TPO), tissue transglutaminase (tTG) and 21-hydroxylase (21-OH) autoantibodies in randomly selected participants with ADM and in age-matched, sex-matched and non-ADM controls with T2D plus controls with NGT. Results Post-normalization, the standardized prevalence rate of ADM was 6.0% (95% confidence interval [CI] 5.3-6.8) in initially non-insulin-requiring participants with ADM, corresponding to six million adults in China, in whom adjusted antibody positivity was: TPO autoantibodies 16.3% (95% CI 10.8-21.8), tTG autoantibodies 2.1% (95% CI 0.0-4.2), and 21-OH autoantibodies 1.8% (95% CI -0.2 to 3.8). Those participants with ADM who were GAD autoantibody-positive had high risk of TPO autoantibody positivity (odds ratio [OR] 2.39, P = 0.0031) and tTG autoantibody positivity (OR 6.98, P = 0.027), while those positive for IA2 autoantibodies had a high risk of tTG autoantibody positivity (OR 19.05, P = 0.001). Conclusions A proportion of people with phenotype of T2D in China have ADM, with diabetes-associated autoantibodies, and may be at risk of developing other organ-specific autoimmune diseases; therefore, it may be clinically relevant to consider screening such Chinese populations.

Summary Generation of functional β cells from pluripotent sources would accelerate diagnostic and therapeutic applications for diabetes research and therapy. However, it has been challenging to generate competent β cells with dynamic insulin-secretory capacity to glucose and incretin stimulations. We introduced transcription factors, critical for β-cell development and function, in differentiating human induced pluripotent stem cells (PSCs) and assessed the impact on the functionality of derived β-cell (psBC) progeny. A perifusion system revealed stepwise transduction of the PDX1, NEUROG3, and MAFA triad (PNM) enabled in vitro generation of psBCs with glucose and GLP-1 responsiveness within 3 weeks. PNM transduction upregulated genes associated with glucose sensing, insulin secretion, and β-cell maturation. In recipient diabetic mice, PNM-transduced psBCs showed glucose-responsive insulin secretion as early as 1 week post transplantation. Thus, enhanced pre-emptive β-cell specification of PSCs by PNM drives generation of glucose- and incretin-responsive psBCs in vitro, offering a competent tissue-primed biotherapy., Highlights • PNM transduction facilitates generation of glucose- and GLP1-responsive β cells • Stepwise PNM improves glucose sensing, insulin secretion, and β-cell maturation • PNM-transduced psBCs showed glucose-responsive insulin secretion in diabetic mice, In this article, Ikeda Yasuhiro and colleagues show that stepwise transduction of the triad of transcription factors PDX1, NEUROG3, and MAFA (PNM) enabled in vitro generation of glucose- and GLP-1-responsive β cells from iPSCs within 3 weeks. PNM transduction improves glucose sensing, insulin secretion, and β-cell maturation of generated cells. PNM-transduced β cells showed glucose-responsive insulin secretion as early as 1 week post transplantation in diabetic mice.

Background Few studies on anastomotic condition after rectal-cancer resection and its effect on anastomotic leakage (AL) are available up to now. This study aimed to investigate potential radiation-induced injury left on surgical margins of anterior resection after neoadjuvant chemoradiotherapy (nCRT) and its association with AL. Methods We retrospectively identified 161 consecutive patients who underwent anterior resection with nCRT, neoadjuvant chemotherapy without radiation (nCT) or no neoadjuvant therapy between 2014 and 2015. Tissue samples of resection margins were assessed using a specific histopathological score and microvessel density in submucosa. Propensity score matching was used to balance the baseline characteristics. Association between AL and histopathological features was analysed. Results AL occurred in 13 of 54 patients undergoing nCRT, 5 of 48 patients undergoing nCT and 7 of 59 patients without neoadjuvant therapy. Comparisons after matching showed median (range) histopathological scores as follows: 3 (0–8) vs 0 (0–3) vs 0 (0–2) for the proximal margin (P

To confirm the effect of melatonin on 3-nitropropionic acid (3-NP)-induced striatal interneuron injury in rats, behavioral test, histology, immunohistochemistry and Western blotting were respectively used to characterize the behavioral changes of experimental animals in motor and cognition, the morphological changes of striatal interneurons and the expression level of protein markers induced by 3-NP. The results showed that (1) 3-NP induced dysfunction of experimental animals in movement, motor coordination and cognition could be relieved by melatonin treatment; (2) The 3-NP-induced lesion area was unvaryingly in dorsolateral striatum, with almost all neuronal loss in the lesion core, however, lots of neurons survived after melatonin treatment; (3) Immunohistochemical staining of the four interneuron types (parvalbuminergic, cholinergic, calretinergic, and neuropeptide Y-neuronal nitric oxide synthase co-containing) showed that, in the lesion core of 3-NP group, loss of the four interneuron types was obvious, but in transition zone, the processes and varicosities of calretinergic, and neuropeptide Y-neuronal nitric oxide synthase co-containing interneurons increased significantly. Melatonin treatment reduced the loss of the four interneuron types in the lesion core, and inhibited the increase of processes and varicosities in the transition zone; (4) Consistent with above results, the expression level of five interneuron protein markers were significantly increased in the striatum after melatonin treatment. Notably, in both the transition zone and the lesion core induced by 3-NP, TUNEL-positive cells were detected, but decreased significantly after melatonin treatment. The present results indicate that melatonin effectively protects the striatal neurons against the injury induced by 3-NP in rats.

The glutamatergic projection from the cerebral cortex and the thalamus extensively innervates the neostriatal neurons. However, some conflicts in the published literatures about cortical and thalamic intrastriatal synaptic terminals still need to be resolved. The present study intends to further elucidate the morphological characteristics of these two types of the terminals and their neurons.The corticostriatal and thalamostriatal terminals were immunolabeled for vesicular glutamate transporter type 1 (VGluT1) and 2 (VGluT2), respectively, and their neurons were retrograde labeled by biotinylated dextran amine 3,000 molecular weight (BDA3k) injection into the dorsolateral striatum of rats. The characteristics of the corticostriatal and thalamostriatal terminals were observed at the LM and EM levels, and the data were statistically analyzed with SPSS10.0 software.We observed that 63.53% of VGluT1+ terminals synapsed on dendritic spines, which was different from VGluT2+ terminals with the equal percentage of synapses on spines and dendrites (14.88 and 17.86%, respectively). Notably, VGluT1+ axospinous synaptic terminals were remarkably larger than VGluT2+ axospinous synaptic terminals. Terminal size-frequency distribution analysis showed that VGluT1+ terminals were within the size ranges of 0.4-0.5 and 0.8-0.9 μm, and VGluT2+ terminals were in the ranges of 0.4-0.5 and 0.6-0.7 μm. Perforated-postsynaptic densities (-PSDs) were more frequently found in VGluT1+ axospinous synaptic terminals than in VGluT2+ axospinous terminals. Furthermore, BDA3k-labeled corticostrital neurons were larger in perikaryal diameter than the thalamostriatal neurons, and they were also categorized as the two main populations based on their size-frequency distribution.The morphological characteristics of corticostriatal and thalamostriatal terminals and neurons have implications for understanding the roles of synaptic plasticity in adaptive motor control by the basal ganglia, and they have facilitations for understanding the complexities of basal ganglia function.

Oxidative stress is closely involved in neurodegenerative diseases. The present study aimed to examine the effect of anti-oxidant DHM (dihydromyricetin) on 3NP (3-nitropropionic acid) -induced behavioral deficits of experimental rats and striatal histopathological injury by using behavioral, imaging, biochemistry, histochemistry and molecular biology technologies. The experimental results showed that both motor dysfunctions and learning and memory impairments induced by 3NP were significantly reduced after DHM treatment. 3NP-induced striatal metabolic abnormality was also remarkably improved by DHM treatment, showed as the increased glucose metabolism in PET/CT scan, decreased MDA (malondialdehyde) and increased SOD (superoxide dismutase) activity in enzyme histochemical staining. In addition, the cell apoptosis was evidently detected in the striatum of the 3NP group, while in the 3NP + DHM group, the number of apoptotic cells was remarkably reduced. 3NP treatment obviously induced down-regulation of Bcl-2, and up-regulations of Bax and Cleaved Caspase-3, while these changes were significantly reversed by DHM treatment. The present results suggested that DHM showed its protective effect by anti-oxidant and anti-apoptosis mechanisms.

Background In the China National Diabetes and Metabolic Disorders Study, age-standardised prevalence of total diabetes was 9·7%, accounting for 92·4 million adults, in China in 2010. Using clinical data and serum samples from this study, we aimed to investigate the prevalence of several organ-specific autoantibodies in patients with phenotypic type 2 diabetes. Methods This study included a nationally representative sample of 46 239 adults aged 20 years or older from 14 provinces and municipalities. After an overnight fast, participants had an oral glucose tolerance test. Previously diagnosed diabetes was self-reported. We randomly selected 4671 patients with diabetes and 1002 individuals with normal glucose tolerance as controls. Participants were screened centrally for glutamic acid decarboxylase autoantibodies (GAD-Ab), protein tyrosine phosphatase-like protein autoantibodies (IA2-Ab), and zinc transporter isoform 8 autoantibodies (ZnT8-Ab) using validated methods of the Islets Autoantibody Standardization Program 2012. We then assayed for thyroid peroxidase autoantibodies (TPO-Ab), tissue transglutaminase autoantibodies (tTG-Ab), and 21-hydroxylase autoantibodies (21-OH-Ab) in patients with islet autoantibody-positive diabetes (n=175), in randomly selected age-matched and sex-matched patients with islet autoantibody-negative diabetes (n=3052 for TPO-Ab, n=525 for tTG-Ab, and n=525 for 21-OH-Ab), and in individuals with normal glucose tolerance (n=954 for TPO-Ab, n=200 for tTG-Ab, and n=200 for 21-OH-Ab). We used multiple logistic regression analysis to determine the risk factor of each autoantibody. Ethics approval was granted by the Second Xiangya Hospital of Central South University, and all participants provided written informed consent. Findings After adjustment for age and sex using results from the national population census, the standardised prevalence of autoimmune diabetes was 6·0% in all patients with diabetes, which corresponded to about 6 million adults in China. In patients with diabetes, the adjusted antibody positivity was 3·6% for GAD-Ab, 1·2% for IA2-Ab, and 1·5% for ZnT8-Ab. In those with autoimmune diabetes, the adjusted antibody positivity was 16·3% for TPO-Ab, 2·1% for tTG-Ab, and 1·8% for 21-OH-Ab. In patients with islet autoantibody-negative diabetes, the adjusted positivity was 10·2% for TPO-Ab, 0·3% for tTG-Ab, and 0·5% for 21-OH-Ab. In Chinese adults with autoimmune diabetes, those positive for GAD-Ab had a high risk for TPO-Ab positivity (odds ratio 2·39, 95% CI 1·35–4·24; p=0·0031) and tTG-Ab positivity (6·983, 1·24–39·22; p=0·027), and those positive for IA2-Ab had a high risk for tTG-Ab positivity (19·05, 3·14–115·61; p=0·0010). Patients positive for TPO-Ab were at high risk of being positive for 21-OHAb (7·48, 1·44–38·74; p=0·016) and vice versa (7·21, 1·35–38·45; p=0·021). Of note, women had a high risk for TPO-Ab positivity compared with men (3·83, 2·20–6·68; p Interpretation Patients positive for islet autoantibodies have a high risk for other organ-specific autoimmune diseases. Screening of TPO-Ab and tTG-Ab in Chinese individuals with autoimmune diabetes should be considered a priority. Funding National Key Technology R&D Program (2012BAI02B04), the Grant of National Clinical Research Center for Metabolic Diseases (2013BAI09B12), the Program for Changjiang Scholars and Innovative Research Team in University (IRT1195), and the Key Project of Chinese Ministry of Education (113050A).

The CRISPR/Cas9 and guide RNA system has emerged as a promising genome editing platform, due to its simplicity, versatility and efficiency, compared to prior techniques for genome modification. Although the original Cas9 gene is too long to be packaged by AAV vectors, Dr. Zhang and colleagues have identified a shorter Cas9 protein from S. aureus (saCas9), which allows generation of a single AAV vector carrying both saCas9 and a guide RNA. Previously, we and others have demonstrated that AAV serotypes 8 and 9-based vectors are highly pancreas-tropic. Our long-term goal is to develop efficient beta-cell- and acinar-cell-targeted gene editing systems based on the AAV-saCas9 system. We first assessed on-target effects of the AAV-saCas-gRNA vector system in vitro. We generated two GFP sequence-targeted gRNA vectors, and tested their genome editing effects using two HT1080 cell lines carrying a single copy of GFP. Infection of HT1080 lines with an multiplicity of infection (MOI) of 2×104 resulted in 40-55% GFP knockout by two AAV-gRNA constructs. Gene editing efficiency was comparable in two HT1080 lines, and dose-dependent. Consecutive, multiple high-dose infections for 3 days achieved up to 80% GFP knockdown. To test whether a subset of cells are more resistant to the AAV-saCas9-mediated genome editing, we sorted the remaining 20% of GFP-positive cells and re-infected recovered GFP-positive cells with the two saCas9 vectors. Our data showed those cells were equally permissive to the saCas9 vectors, when compared to their parental line. To assess the off-target effects of the AAV-saCas9-gRNA system, we compared the exome sequences of the vector-treated cells. Our preliminary result identified multiple off-target deletions upon AAV-saCas9 treatments. We are currently analyzing whether those off-target effects were due to their similarity to the GFP targeting gRNA sequences, or independent from the gRNA sequences. We then tested the feasibility of the AAV-saCas9-gRNA vector system for beta-cell-targeted gene editing in vivo. We generated AAV8 vectors carrying Pdx1-targeting gRNA, with saCas9 driven by CMV promoter. Single intraperitoneal injection of AAV saCas9 vectors led to efficient knockout of Pdx1 expression in the pancreatic beta-cells. To limit potential off-target effects, we need to restrict saCas9 expression in beta-cells. However, our original beta-cell targeted vectors carry a relatively long, 1.1kb insulin promoter sequence. We therefore optimized the insulin promoter sequence and determined the minimum, essential promoter sequence, which facilitates robust transgene expression with high tissue-specificity. We are in the process of testing the AAV-saCas9-gRNA vectors with the optimized insulin promoter sequence for in vivo beta-cell gene editing.

Maize is not only a staple food crop but also an important raw material for feed and industry; however, the threat of weeds leads to a serious decline in its output and quality. The G2-aroA gene confers glyphosate herbicide tolerance to crops. In this study, the food safety of genetically modified (GM), glyphosate-tolerant maize with the G2-aroA gene was evaluated in a 90-day feeding study in Sprague-Dawley (SD) rats. Maize grain from GM or non-GM isogenic control lines were separately formulated into rodent diets at concentrations of 12.5% (low level), 25% (middle level), and 50% (high level). An additional group of rats were fed a commercialized diet as a control. The toxicological response variables, including body weights, food consumption, serum biochemistry, hematology, and absolute and relative organ weights, were compared between rats fed GM maize and those fed non-GM maize after consumption of test diets for 90days. In addition, gross and microscopic pathology were conducted among treatment groups. No adverse effects related to the consumption of GM maize were detected in the subchronic feeding study. These results indicated that the GM glyphosate-tolerant maize was as safe and nutritious as conventional maize.

Background The present study sought to investigate pathologic changes in tendon, expression of basic fibroblast growth factor (bFGF) and collagen type I, and effects of safflower yellow (SY) on the process of tendon injury-repair. Materials and Methods A tendon injury-repair model was used, and stereology, biomechanics, and immunohistochemistry were employed to assess the benefits of local application of SY for the repair. In this model, the flexor digitorum profundus muscle tendon of the third digit was transected bilaterally, and the transected ends sutured. Data were analyzed with SPSS ver. 10.0 software (SPSS Inc., Chicago, IL). Results The adhesion to surrounding tissues and tensile strength gradually increased after the injury and repair in control (no-SY) tendons, and were significantly greater by the sixth wk than any other time. In the SY tendons, adhesion was significantly lower, and tensile strength significantly higher than in no-SY tendons at the same post-injury-suture time points. An inflammatory reaction was observed in the injury-repair areas of the tendon by the end of first wk post-injury-suture, and reached its peak by the end of second wk. The inflammatory reaction was significantly less in SY tendons than in controls. Immunostaining for bFGF occurred in the tendon injury-repair areas by the end of first wk, and the number of bFGF positive cells reached a peak by the end of second wk, with a greater abundance in SY than control tendons from the second to sixth wk. Expression of collagen type I protein was observed in the injury-repair areas as well, coincident with bFGF, and was remarkably higher in SY than in controls. Conclusions Tendon adhesion and tensile strength increased with time post-injury-suture repair, as did expression of bFGF and collagen type I protein in the injured area. SY enhanced expression of bFGF and collagen type I protein, enhanced the tensile strength of the injured tendon, and alleviated the injured tendon adhesion and inflammatory reaction. The results indicated that SY promoted the repair of injured tendon by up-regulating expression of bFGF and collagen type I protein.

Histology, immunohistochemistry, and Western blotting were used to characterize the changes in morphology, distribution pattern, and marker protein expression of striatal interneurons in the transition zone of striatal injury induced by 3-NP. The 3-NP treatment in rats yielded movement, motor coordination, and cognitive dysfunction. The 3-NP-induced lesion core was unvaryingly in the dorsolateral striatum, with a transition zone of lesser damage around the lesion core, in which medium-sized neurons were significantly decreased in abundance, but larger neurons survived. In both the transition zone and the lesion core, many TUNELpositive cells negative for the interneuron markers were detected, indicating widespread projection neuron death. Immunohistochemical staining for the four interneuron types (parvalbuminergic, cholinergic, calretinergic, and neuropeptide Y–neuronal nitric oxide synthase cocontaining) showed that few immunolabeled interneurons were observed in the lesion core, but interneuron perikarya showed no evident loss in the transition zone. Consistently with this, Western blotting showed that the five interneuron protein markers were significantly decreased in the striatum after 3-NP treatment. Transition-zone calretinergic and neuropeptide Y– neuronal nitric oxide synthase-cocontaining interneurons, however, possessed more processes and varicosities than normal. These results show that, although striatal interneurons survive in the transition zone after 3-NP-mediated striatal injury, they have enhanced marker protein levels in their processes. V C 2011 Wiley

Inflammatory response after middle cerebral artery occlusion (MCAO) has been a focus of research recently, but the effect of inflammatory cells on ischemic neurons remains unclear. In order to study the effect of the inflammatory reaction on brain ischemic injury, we observed the morphology, number and distribution of CD3-, CD8-, ED1- and ED2-positive cells systematically in the caudate-putamen of rats in a MCAO model. The present results show that all four types of inflammatory cells first infiltrated the ischemic penumbra and then migrated into the center of the ischemic area, but the morphological changes and infiltration processes differed significantly; the infiltration of CD3- and CD8-positive cells into the ischemic area started at 3 days postischemia, and their number peaked at 1 week; however, although ED1- and ED2-positive cells were also observed at 3 days after ischemia, they reached their maximum number at 2 and 4 weeks, respectively. Moreover, ED1-and ED2-positive cells showed evident hyperplasia and hypertrophy in morphology. Our results also showed that the response of CD3-, CD8-, ED1- and ED2-positive cells in the ischemic area and the pathological changes in ischemic brain tissue could be inhibited by cyclosporine A. The results suggest that the infiltration and reaction of inflammatory cells are involved in the pathological process of ischemic brain injury.