Mendelian randomization analysis elucidates the causal relationship between celiac disease and the risk of thyroid dysfunction.

The link between celiac disease (CeD) and thyroid dysfunction has been investigated. However, it is uncertain if CeD is causally linked to thyroid dysfunction. A 2-sample Mendelian randomization study was conducted to ascertain the causal connection between CeD and thyroid dysfunction. Using data from the FinnGen Consortium, a 2-sample Mendelian randomization study was conducted to look at the connection between thyroid dysfunction and CeD. Another replication of the data from the UK Biobank was subsequently performed to confirm our findings. Furthermore, a sequence of sensitivity analyses was performed. The inverse variance weighting technique demonstrates that genetically determined CeD is substantially linked with hypothyroidism, thyrotoxicosis, Graves' disease, and free thyroxine. However, no significant associations were found between CeD and thyroid-stimulating hormone or thyroiditis. Moreover, we achieve the same results in duplicate datasets, which increases the reliability of our findings. This study suggests that CeD and thyroid dysfunction are linked, and it gives theoretical support and new ways of thinking about how to diagnose and treat both conditions.

Research Advances on Stem Cell-Derived Extracellular Vesicles Promoting the Reconstruction of Alveolar Bone through RANKL/RANK/OPG Pathway.

Periodontal bone tissue defects and bone shortages are the most familiar and troublesome clinical problems in the oral cavity. Stem cell-derived extracellular vesicles (SC-EVs) have biological properties similar to their sources, and they could be a promising acellular therapy to assist with periodontal osteogenesis. In the course of alveolar bone remodeling, the RANKL/RANK/OPG signaling pathway is an important pathway involved in bone metabolism. This article summarizes the experimental studies of SC-EVs applied for the therapy of periodontal osteogenesis recently and explores the role of the RANKL/RANK/OPG pathway in their mechanism of action. Their unique patterns will open a new field of vision for people, and they will help to advance a possible future clinical treatment.

Perspective on advanced directives among older adults in Shanghai: A qualitative study.

Objective: This study aimed to explore the perception on advanced directives (ADs) among older adults in Shanghai. Methods: Through purposive sampling, 15 older adults with rich life experiences who were willing to share perceptions and experiences of ADs participated in this study. Face-to-face semi-structured interviews were conducted to collect the qualitative data. Thematic content analysis was applied to analyze the data. Results: Five themes have been identified: low awareness but high acceptance of ADs; pursuing natural and peaceful sunset life; ambiguous attitude on medical autonomy; being irrational facing patients' dying and death issues; positive about implementing ADs in China. Conclusion: It is possible and feasible to implement ADs in older adults. Death education and compromised medical autonomy may be needed in the Chinese context as the foundation. The elder's understanding, willingness and worries about ADs should be fully revealed. Diverse approaches should be applied to introduce and interpret ADs to older adults continuously.

Sequentially releasing self-healing hydrogel fabricated with TGFß3-microspheres and bFGF to facilitate rat alveolar bone defect repair.

Resorption and loss of alveolar bone leads to oral dysfunction and loss of natural or implant teeth. Biomimetic delivery of growth factors based on stem cell recruitment and osteogenic differentiation, as the key steps in natural alveolar bone regenerative process, has been an area of intense research in recent years. A mesoporous self-healing hydrogel (DFH) with basic fibroblast growth factor (bFGF) entrapment and transforming growth factor ß3 (TGFß3) - loaded chitosan microspheres (CMs) was developed. The formulation was optimized by multiple tests of self-healing, in-bottle inversion, SEM, rheological, swelling rate and in vitro degradation. In vitro tubule formation assays, cell migration assays, and osteogenic differentiation assays confirmed the ability of DFH to promote blood vessels, recruit stem cells, and promote osteogenic differentiation. The optimum DFH formula is 0.05 ml 4Arm-PEG-DF (20%) added to 1 ml CsGlu (2%) containing bFGF (80 ng) and TGFß3-microspheres (5 mg). The results of in vitro release studied by Elisa kit, indicated an 95% release of bFGF in 7 d and long-term sustained release of TGFß3. For alveolar defects rat models, the expression levels of CD29 and CD45, the bone volume fraction, trabecular number, and trabecular thickness of new bone monitored by Micro-CT in DFH treatment groups were significantly higher than others (*P < 0.05, vs Model). HE and Masson staining show the same results. In conclusion, DFH is a design of bionic alveolar remodelling microenvironment, that is in early time microvessels formed by bFGF provide nutritious to recruited endogenous stem cells, then TGFß3 slowly released speed up the process of new bones formation to common facilitate rat alveolar defect repair. The DFH with higher regenerative efficiency dovetails nicely with great demand due to the requirement of complicated biological processes.

Transforming Growth Factor-ß3/Recombinant Human-like Collagen/Chitosan Freeze-Dried Sponge Primed With Human Periodontal Ligament Stem Cells Promotes Bone Regeneration in Calvarial Defect Rats.

Patients with a skull defect are at risk of developing cerebrospinal fluid leakage and ascending bacterial meningitis at >10% per year. However, treatment with stem cells has brought great hope to large-area cranial defects. Having found that transforming growth factor (TGF)-ß3 can promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs), we designed a hybrid TGF-ß3/recombinant human-like collagen recombinant human collagen/chitosan (CS) freeze-dried sponge (TRFS) loading hPDLSCs (TRFS-h) to repair skull defects in rats. CFS with 2% CS was selected based on the swelling degree, water absorption, and moisture retention. The CS freeze-dried sponge (CFS) formed a porous three-dimensional structure, as observed by scanning electron microscopy. In addition, cytotoxicity experiments and calcein-AM/PI staining showed that TRFS had a good cellular compatibility and could be degraded completely at 90 days in the implantation site. Furthermore, bone healing was evaluated using micro-computed tomography in rat skull defect models. The bone volume and bone volume fraction were higher in TRFS loaded with hPDLSCs (TRFS-h) group than in the controls (p < 0.01, vs. CFS or TRFS alone). The immunohistochemical results indicated that the expression of Runx2, BMP-2, and collagen-1 (COL Ⅰ) in cells surrounding bone defects in the experimental group was higher than those in the other groups (p < 0.01, vs. CFS or TRFS alone). Taken together, hPDLSCs could proliferate and undergo osteogenic differentiation in TRFS (p < 0.05), and TRFS-h accelerated bone repair in calvarial defect rats. Our research revealed that hPDLSCs could function as seeded cells for skull injury, and their osteogenic differentiation could be accelerated by TGF-ß3. This represents an effective therapeutic strategy for restoring traumatic defects of the skull.

Sulfonated chitosan oligosaccharide alleviates the inhibitory effect of basic fibroblast growth factor on osteogenic differentiation of human periodontal ligament stem cells.

BACKGROUND: Periodontal ligament stem cells (PDLSCs) play an essential role in periodontal tissue repair. Basic fibroblast growth factor (bFGF) has been used in the clinical treatment of periodontal disease. However, studies have shown that bFGF inhibits the osteogenic differentiation of PDLSCs, which is not conducive to alveolar bone repair. Sulfonated chitosan oligosaccharide (SCOS), a heparan-like compound, can maintain the conformation of bFGF and promote its proliferation activity. This study investigated the effects of bFGF in combination with SCOS on the osteogenic differentiation of hPDLSCs. METHODS: hPDLSCs were isolated from healthy human periodontal ligament and identified by flow cytometry and immunofluorescence. The affinity between SCOS and bFGF was analyzed by surface plasmon resonance. Changes in osteogenic differentiation by combination of bFGF with SCOS were analyzed by alkaline phosphatase activity assay, Sirius Red staining, and Alizarin Red staining. Expression of genes and proteins was investigated by western blotting and reverse transcription-quantitative PCR. RESULTS: Extracted hPDLSCs were mesenchymal stem cells with pluripotent differentiation potential. SCOS exhibited an affinity for bFGF. bFGF (20 ng/mL) promoted the proliferation of hPDLSCs, but inhibited their osteogenic differentiation. SCOS alleviated the inhibitory effect of bFGF on the osteogenic differentiation of hPDLSCs. CONCLUSIONS: SCOS can reduce the inhibitory effect of bFGF on the osteogenic differentiation of hPDLSCs. This study provides evidence for the clinical use of bFGF to repair periodontal tissue.

Transforming Growth Factor-ß3/Chitosan Sponge (TGF-ß3/CS) Facilitates Osteogenic Differentiation of Human Periodontal Ligament Stem Cells.

Periodontal disease is the main reason for tooth loss in adults. Tissue engineering and regenerative medicine are advanced technologies used to manage soft and hard tissue defects caused by periodontal disease. We developed a transforming growth factor-ß3/chitosan sponge (TGF-ß3/CS) to repair periodontal soft and hard tissue defects. We investigated the proliferation and osteogenic differentiation behaviors of primary human periodontal ligament stem cells (hPDLSCs) to determine the bioactivity and potential application of TGF-ß3 in periodontal disease. We employed calcein-AM/propidium iodide (PI) double labeling or cell membranes (CM)-Dil labeling coupled with fluorescence microscopy to trace the survival and function of cells after implantation in vitro and in vivo. The mineralization of osteogenically differentiated hPDLSCs was confirmed by measuring alkaline phosphatase (ALP) activity and calcium content. The levels of COL I, ALP, TGF-ßRI, TGF-ßRII, and Pp38/t-p38 were assessed by western blotting to explore the mechanism of bone repair prompted by TGF-ß3. When hPDLSCs were implanted with various concentrations of TGF-ß3/CS (62.5-500 ng/mL), ALP activity was the highest in the TGF-ß3 (250 ng/mL) group after 7 d (p < 0.05 vs. control). The calcium content in each group was increased significantly after 21 and 28 d (p < 0.001 vs. control). The optimal result was achieved by the TGF-ß3 (500 ng/mL) group. These results showed that TGF-ß3/CS promotes osteogenic differentiation of hPDLSCs, which may involve the p38 mitogen-activated protein kinase (MAPK) signaling pathway. TGF-ß3/CS has the potential for application in the repair of incomplete alveolar bone defects.

High-Indexed PtNi Alloy Skin Spiraled on Pd Nanowires for Highly Efficient Oxygen Reduction Reaction Catalysis.

The catalytic performance of Pt-based catalysts for oxygen reduction reactions (ORR) can generally be enhanced by constructing high-index exposed facets (HIFs). However, the synthesis of Pt alloyed high-index skins on 1D non-Pt surfaces to further improve Pt utilization and stability remains a fundamental challenge for practical nanocrystals. In this work, Pd nanowires (NWs) are selected as a rational medium to facilitate the epitaxial growth of Pt and Ni. Based on the different nucleation and growth habits of Pt and Ni, a continuous PtNi alloy skin bounded with HIFs spiraled on a Pd core can be obtained. Here, the as-prepared helical Pd@PtNi NWs possess high HIF densities, low Pt contents, and optimized oxygen adsorption energies, demonstrating an enhanced ORR mass activity of 1.75 A mgPt -1 and a specific activity of 3.18 mA cm-2 , which are 10 times and 12 times higher than commercial Pt/C catalysts, respectively. In addition, the 1D nanostructure enables the catalyst to be highly stable after 30 000 potential sweeping cycles. This work successfully extends bulky high-indexed Pt alloys to core-shell nanostructures with the design of a new, highly efficient and stable Pt-based catalyst for fuel cells.

Improving the protective effects of aFGF for peripheral nerve injury repair using sulfated chitooligosaccharides.

Injury to the peripheral nerves can result in temporary or life-long neuronal dysfunction and subsequent economic or social disability. Acidic fibroblast growth factor (aFGF) promotes the growth and survival of neurons and is a possible treatment for peripheral nerve injury. Yet, the actual therapeutic utility of aFGF is limited by its short half-life and instability in vivo. In the present study, we prepared sulfated chitooligosaccharides (SCOS), which have heparin-like properties, to improve the bioactivity of aFGF. We investigated the protective effects of SCOS with or without aFGF on RSC96 cells exposed to Na2S2O4 hypoxia/reoxygenation injury. Cell viability was measured by MTT assay and cytotoxicity induced by Na2S2O4 was assessed by lactate dehydrogenase (LDH) release into the culture medium. Pretreatment with aFGF and SCOS dramatically decreased LDH release after injury compared to pretreatment with aFGF or SCOS alone. We subsequently prepared an aFGF/SCOS thermo-sensitive hydrogel with poloxamer and examined its effects in vivo. Paw withdrawal thresholds and thermal withdrawal latencies were measured in rats with sciatic nerve injury. Local injection of the aFGF/SCOS hydrogels (aFGF: 40, 80 µg/kg) increased the efficiency of sciatic nerve repair compared to aFGF (80 µg/kg) hydrogel alone. Especially aFGF/SCOS thermo-sensitive hydrogel decreased paw withdrawal thresholds from 117.75 ± 8.38 (g, 4 d) to 65.74 ± 3.39 (g, 10 d), but aFGF alone group were 140.58 ± 27.54 (g, 4 d) to 89.12 ± 5.60 (g, 10 d) (aFGF dose was 80 µg/kg, P < 0.05, n = 8). The thermal withdrawal latencies decreased from 11.61 ± 2.26 (s, 4 d) to 2.37 ±0.67 (s, 10 d). However, aFGF alone group were from 17.69 ± 1.47 (s, 4 d) to 4.65 ± 1.73 (s, 10 d) (P < 0.05, n = 8). Furthermore, the aFGF/SCOS hydrogels also exhibited good biocompatibility in mice. In summary, SCOS improved the protective effects of aFGF in RSC96 cells injured with Na2S2O4 and increased the efficiency of nerve repair and recovery of function in rats with sciatic nerve injury. These findings pave an avenue for the development of novel prophylactic and therapeutic strategies for peripheral nerve injury.

Research progress on the factors influencing the biological characteristics of dental pulp stem cells / 口腔疾病防治

@#Dental pulp stem cells (DPSCs) are mesenchymal stem cells derived from dental pulp tissue with self-renewal, high proliferative capacity and multidirectional differentiation potential. Under appropriate induction conditions, DPSCs can be differentiated into various types of cells, such as osteoblasts, odontoblasts, chondrocytes, adipocytes, and neuronal cells. DPSCs have been gradually applied to clinical trials and preclinical studies and are important seed cells in the field of periodontal tissue engineering and regenerative medicine. In this paper, the factors affecting the biological characteristics of DPSCs are reviewed together with a review of recent literature published worldwide. The results of the literature review show that the biological characteristics of DPSCs can be influenced by many factors, such as tissue source, culture method, environment and induction conditions, which has guiding significance for research and applications of DPSCs.

Research advances in the correlations between periodontal disease and autophagy / 口腔疾病防治

@#Periodontal disease is a common and frequently occurring oral disease, with numerous investigations on periodontitis etiology and treatment. The development of periodontal disease is affected by many factors, including local stimulation and systemic risk factors, and dental plaque biofilm is regarded as an initial factor. The molecular mechanisms of how dental plaque biofilm and other risk factors regulate the development of periodontal disease remain unclear. Autophagy is a ubiquitous self-regulating mechanism of eukaryotes and a process of cell self-phagocytosis and digestion. Autophagy is a complex process, involving a variety of cellular and tissue activities, including cellular stress, clearance of endogenous and exogenous cellular components, development, aging and cancer. Recent studies have shown that autophagy is closely related to inflammation. This paper addresses recent research advances in understanding the correlations between periodontal disease and autophagy to provide a reference in the study of new methods to treat periodontal disease from the perspective of autophagy in the future.

Pulmonary Function and Retrobulbar Hemodynamics in Subjects With Type 2 Diabetes Mellitus.

BACKGROUND: The primary goals of this study were to evaluate early changes in pulmonary function and retrobulbar hemodynamics and to examine the correlation between these parameters in patients with type 2 diabetes during the preclinical stages of diabetic retinopathy. METHODS: For the single-time point measurements, 63 subjects with type 2 diabetes without diabetic retinopathy (diabetes group) and 32 healthy subjects (control group) were selected to evaluate any early changes in pulmonary function and retrobulbar hemodynamics and to examine the correlation between these parameters. In the longitudinal follow-up study, 32 subjects who were newly diagnosed with type 2 diabetes were divided into 2 groups according to their resistivity index (≤0.7 and >0.7). Early changes in pulmonary function and retrobulbar hemodynamics were studied in these groups and compared with the previous values. RESULTS: For the single-time point measurements, the fasting plasma glucose, 2-h postprandial blood glucose, glycosylated hemoglobin A1c, total cholesterol, low-density lipoprotein cholesterol, and triglyceride levels as well as the pulmonary function parameters were significantly higher in the diabetes group than in the control group. The pulmonary function parameters were negatively and significantly correlated with glycosylated hemoglobin A1c and the duration of diabetes. The retrobulbar hemodynamics were positively correlated with glycosylated hemoglobin A1c and diabetes duration; in contrast, the correlation between retrobulbar hemodynamics and glycosylated hemoglobin A1c. In the longitudinal follow-up study, the pulmonary function of the 2 groups categorized by their resistivity index levels indicated that subjects with resistivity index levels ≤0.7 showed significantly better pulmonary function, and the pulmonary function of this group showed improvement and a significantly smaller decrease. The incidence of diabetic retinopathy in the group with resistivity index levels ≤0.7 (9 of 22, 40.9%) was significantly lower than that in the group with resistivity index levels >0.7. CONCLUSIONS: Pulmonary function and retrobulbar hemodynamics changed during the preclinical stages of diabetic retinopathy. Regulating glycemia may improve retrobulbar hemodynamics in the retrobulbar arteries (ie, central retinal artery, posterior ciliary artery, and arteria ophthalmica). By detecting the retrobulbar resistivity index and the levels of glycosylated hemoglobin A1c, we could predict future changes in pulmonary function during the preclinical stages of diabetic retinopathy as well as the degree of retinopathy. (ClinicalTrials.gov registration NCT02774733.).

Progress in relationship between leptin and periodontitis / 口腔疾病防治

@#The in itiation and progression of periodontal disease were reported to be the results of complicated interactions between specific subgingival bacteria and host immuno-inflammatory response. As a part of the immune and defense system mechanisms of the host, leptin may have a protective effect on periodontal tissues. We summarize the latest progress in the relationship between leptin and periodontitis.

The effect of alogliptin on pulmonary function in obese patients with type 2 diabetes inadequately controlled by metformin monotherapy.

BACKGROUND: To observe the effect of alogliptin combined with metformin on pulmonary function in obese patients with type 2 diabetes inadequately controlled by metformin monotherapy (500 mg, bid po, for at least 3 months), and evaluate its efficacy and safety. METHODS: After a 2-week screening period, adult patients (aged 36-72 years) entered a 4-week run-in/stabilization period. Then, patients were randomly assigned to either the intervention group (n = 55) or the control group (n = 50) for 26 weeks. The patients in the control group were given metformin (1000 mg, bid po) and the patients in the intervention group were given metformin (500 mg, bid po) combined with alogliptin (25 mg, qd po). All the patients received counseling about diet and exercise from a nutritionist during run-in and treatment periods.The primary endpoints were the between-group differences in the changes in pulmonary function parameters (vital capacity [VC]%, forced vital capacity [FVC]%, forced expiratory volume in 1 second (FEV1)%, peak expiratory force [PEF]%, maximal voluntary ventilation [MVV]%, total lung capacity [TLC%], forced expiratory volume in 1 second/forced vital capacity [FEV1/FVC%], diffusing capacity for carbon monoxide of lung [DLCO]%, and diffusing capacity for carbon monoxide of lung/unit volume [DLCO/VA%]) between pretherapy and posttreatment. The secondary endpoints were changes from baseline to week 26 in glycosylated hemoglobinA1c (HbA1c), FPG, 2hPG, homeostasis model assessment insulin resistance (HOMA-IR), waist circumference (WC), and BMI. The tertiary endpoints were the changes from baseline to week 26 in blood-fat (total cholesterol [TC], high-density lipoprotein cholesterol [HDL-C], low-density lipoprotein cholesterol [LDL-C], and triglycerides [TG]). The quartus endpoints were the changes from baseline to week 26 in systolic blood pressure (SBP) and diastolic blood pressure (DBP). The 5th endpoints were the changes from baseline to week 26 in oxidative/antioxidative parameters (reactive oxygen species [ROS], malondialdehyde [MDA], superioxide dismutase [SOD], and glutathione peroxidase [GSH-Px]). In addition, safety endpoints were assessed (AEs, clinical laboratory tests, vital signs, and electrocardiographic readings). RESULTS: Eighty-one patients completed our clinical trial: intervention group (n = 44) and control group (n = 37). At week 26, pulmonary function parameters (VC%, FVC%, FEV1%, PEF%, MVV%, TLC%, FEV1/FVC%, DLCO%, and DLCO/VA%) had increased significantly from pretherapy values in both groups (P < 0.05), and the pulmonary function tests were significantly greater (P < 0.05) in intervention group than in controls posttherapy. Pulmonary function (FVC%, FEV1%, PEF%, TLC%, FEV1/FVC%, DLCO%, and DLCO/VA%) was lower in the group with HbA1c levels ≥8.0 at 26 weeks, but VC%, FEV1%, MVV%, and TLC% were not significantly lower (P > 0.05). Pulmonary function parameters were positively correlated with GSH-Px and SOD and negatively correlated with ROS and MDA. Mean declines in HbA1c, FPG, 2hPG, HOMA-IR, and blood-fat (TC, HDL-C, LDL-C, and TG) were significantly greater (P < 0.05) in intervention group compared with the controls, but mean declines in BMI, WC, and BP (SBP, DBP) did not differ significantly between the 2 groups (P > 0.05). SOD and GSH-Px increased more (P < 0.05) in the intervention group, compared with the controls; ROS and MDA declined more (P < 0.05) in intervention group, as compared with the control group. The most common AEs were gastrointestinal events, headaches, skin-related AEs (mostly pruritic events), and hypoglycemia. The incidences of AEs did not differ significantly (P > 0.05) between the 2 groups except for the headache and skin-related adverse events (the incidence of headache was higher in the intervention group than in controls; P < 0.05). No patient died during the study. CONCLUSION: In patients with type 2 diabetes mellitus (T2DM) inadequately controlled by metformin monotherapy, the addition of alogliptin contributed to clinically significant increases in pulmonary function through regulating glycemia and improving the imbalance of the oxidative-related substances in the serum, without increasing the incidence of hypoglycemia, dyslipidemia, dysarteriotony, and any notable increase in body weight.

Improving cell growth and lipid accumulation in green microalgae Chlorella sp. via UV irradiation.

Microalgae with high biomass and high lipid content are the ideal feedstock for biodiesel production. To obtain such microalgae, ultraviolet (UV) irradiation was applied to Chlorella sp. to induce mutagenesis. The growth characteristics, total nitrogen (TN), and biochemical compositions of the control and UV mutation strains were analyzed. Compared to the control strain, the biomass for the UV mutation strain was 7.6 % higher and it presented a higher growth rate. The lipid content of the UV mutation strain showed different levels of increase and reached the maximum value of 28.1 % on day 15. Furthermore, the lipid productivity of the UV mutation strain showed a desired increase. The nitrogen consumption and Acetyl-CoA carboxylase (ACC) activity contributed to the lipid production by UV. All these results indicate that UV mutagenesis is an efficient method to improve probability for using Chlorella sp. as the potential raw material for biodiesel production.

[Determination of organochlorine pesticides in water using solidification of floating organic drop liquid phase microextraction by gas chromatography].

OBJECTIVE: To develop a new method for simultaneous determination of eight organochlorine pesticides (OCPs) in water samples using solidification floating organic drop liquid-phase microextraction (SFO-LPME) combined with gas chromatography-electronic capture detector (GC-ECD). METHODS: The experimental conditions of SFO-LPME were determined with n-Hexadecane as extractant, water samples adjusted to pH 6.0, inonic strength increased by adding 15.0 g/100 mL NaCI. The OCPs were extracted at 55 degrees C for 10 mm and determined with GC-ECD. RESULTS: A good linearity (correlation coefficients > or = 0.996) was obtained for eight target compounds from 5 ng/I. to 100 ng/L. The method detection limits ranged from 0.24 ng/L to 0.78 ng/L. Satisfactory results were achieved with samples of river water, piped water and farmland water, with an average recovery of 76.0%-106.0% and RSD of 3.24%-11.60%. CONCLUSION: The proposed method is rapid, simple and sensitive. It is suitable for hatch analyses of eight organic chlorine pesticides in water samples.

Complete genome analysis of the C4 subgenotype strains of enterovirus 71: predominant recombination C4 viruses persistently circulating in China for 14 years.

Genetic recombination is a well-known phenomenon for enteroviruses. To investigate the genetic characterization and the potential recombination of enterovirus 71 (EV71) circulating in China, we determined the 16 complete genome sequences of EV71 isolated from Hand Foot Mouth Disease (HFMD) patients during the large scale outbreak and non-outbreak years since 1998 in China. The full length genome sequences of 16 Chinese EV71 in present study were aligned with 186 genome sequences of EV71 available from GenBank, including 104 China mainland and 82 international sequences, covering the time period of 1970-2011. The oldest strains of each subgenotype of EV71 and prototype strains of HEV-A were included to do the phylogenetic and Simplot analysis. Phylogenetic analysis indicated that all Chinese strains were clustered into C4 subgenotype of EV71, except for HuB/CHN/2009 clustered into A and Xiamen/CHN/2009 clustered into B5 subgenotype. Most of C4 EV71 were clustered into 2 predominant evolutionary branches: C4b and C4a evolutionary brunches. Our comprehensive recombination analysis showed the evidence of genome recombination of subgenotype C4 (including C4a and C4b) sequences between structural genes from genotype C EV71 and non-structural genes from the prototype strains of CAV16, 14 and 4, but the evidence of intratypic recombination between C4 strains and B subgenotype was not enough strong. This intertypic recombination C4 viruses were first seen in 1998 and became the predominant endemic viruses circulating in China mainland for at least 14 years. A shift between C4a and C4b evolutionary brunches of C4 recombination viruses were observed, and C4a viruses have been associated with large scale nationwide HFMD outbreak with higher morbidity and mortality since 2007.

A novel agonistic anti-human death receptor 5 monoclonal antibody with tumoricidal activity induces caspase- and mitochondrial-dependent apoptosis in human leukemia Jurkat cells.

An agonistic antibody against TNF-related apoptosis-inducing ligand death receptor 5 (DR5) is a practicable candidate drug for antitumor therapy. In this study, a novel murine anti-human DR5 monoclonal antibody, mDRA-6(IgG1-κ), has been generated. This study aimed to explore the caspase-dependent and mitochondrial mechanisms of mDRA-6 in inducing apoptosis in human leukemia Jurkat cells. The apoptotic effects of mDRA-6 on Jurkat cells, which express DR5 on the cell surface, were detected by flow cytometry and western blot after exposure to different doses of mDRA-6 and at fixed doses of mDRA-6 at different times. It was demonstrated that mDRA-6 can induce Jurkat cell apoptosis via caspase- and mitochondrial-dependent pathways. These results indicate that the novel antibody mDRA-6 against DR5 has an antitumor function and may provide a new reagent for tumor therapy.

An emerging recombinant human enterovirus 71 responsible for the 2008 outbreak of hand foot and mouth disease in Fuyang city of China.

Hand, foot and mouth disease (HFMD), a common contagious disease that usually affects children, is normally mild but can have life-threatening manifestations. It can be caused by enteroviruses, particularly Coxsackieviruses and human enterovirus 71 (HEV71) with highly variable clinical manifestations. In the spring of 2008, a large, unprecedented HFMD outbreak in Fuyang city of Anhui province in the central part of southeastern China resulted in a high aggregation of fatal cases. In this study, epidemiologic and clinical investigations, laboratory testing, and genetic analyses were performed to identify the causal pathogen of the outbreak. Of the 6,049 cases reported between 1 March and 9 May of 2008, 3023 (50%) were hospitalized, 353 (5.8%) were severe and 22 (0.36%) were fatal. HEV71 was confirmed as the etiological pathogen of the outbreak. Phylogenetic analyses of entire VP1 capsid protein sequence of 45 Fuyang HEV71 isolates showed that they belong to C4a cluster of the C4 subgenotype. In addition, genetic recombinations were found in the 3D region (RNA-dependent RNA polymerase, a major component of the viral replication complex of the genome) between the Fuyang HEV71 strain and Coxsackievirus A16 (CV-A16), resulting in a recombination virus. In conclusion, an emerging recombinant HEV71 was responsible for the HFMD outbreak in Fuyang City of China, 2008.

Caspase-dependent molecular mechanisms of anti-human DR5 monoclonal antibody mDRA-6 inducing apoptosis of human leukemia Jurkat cells.

BACKGROUND AND OBJECTIVE: Both tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and some monoclonal agonistic antibodies against TRAIL receptors have antitumor activity. We have previously prepared a novel monoclonal agonistic antibody against human death receptor 5 (DR5) and designated it as mDRA-6. This study was to explore the Caspase-dependent molecular mechanisms of mDRA-6 inducing apoptosis of human leukemia Jurkat cells. METHODS: After exposure to different doses of mDRA-6, DNA fragmentation of Jurkat cells was detected by agarose gel electrophoresis, cell proliferation was detected by MTT assay, and cell apoptosis was detected by flow cytometry after Annexin V-FITC/PI double staining. Jurkat cells were further treated with the inhibitors for Caspase-10, -9, -8 and -3. The active cleavage products of Caspase-10, -9, -8, -3 and poly ADP-ribose polymerase (PARP), BH3 interacting domain death agonist (Bid), truncated Bid (tBid) and cytochrome c (Cyto c), were analyzed by western blot. RESULTS: After mDRA-6 treatment, DNA fragmentation was detected in Jurkat cells. mDRA-6 inhibited cell proliferation in a dose-dependent manner. When treated with 2.0 microg/mL mDRA-6, the apoptosis rates of Jurkat cells were 16.2% at 0.25 h, 28.3% at 0.5 h, 69.2% at 1 h and 78.2% at 2 h. Interestingly, the mDRA-6-induced apoptosis was repressed by 77.9% by Caspase-8 inhibitor ZIF, 54.2% by Caspase-3 inhibitor ZDF, and 8.7% by Caspase-9 inhibitor ZLF, but was not repressed by Caspase-10 inhibitor ZAF. After mDRA-6 exposure, the proenzymes of Caspase-8, -9 and -3 were reduced and their active cleavage products were increased along with the increase of exposure time, the cleavage products of PARP were also increased, Bid was degraded to tBid, and an abundance of Cyto c was released from mitochondria, but the proenzyme of Caspase-10 showed no change and no cleavage products of Caspase-10 were detectable. CONCLUSION: mDRA-6 can induce apoptosis of Jurkat cells via the Caspase-dependent and mitochondrial pathways.