Engineered tissue geometry and Plakophilin-2 regulate electrophysiology of human iPSC-derived cardiomyocytes.

Engineered heart tissues have been created to study cardiac biology and disease in a setting that more closely mimics in vivo heart muscle than 2D monolayer culture. Previously published studies suggest that geometrically anisotropic micro-environments are crucial for inducing "in vivo like" physiology from immature cardiomyocytes. We hypothesized that the degree of cardiomyocyte alignment and prestress within engineered tissues is regulated by tissue geometry and, subsequently, drives electrophysiological development. Thus, we studied the effects of tissue geometry on electrophysiology of micro-heart muscle arrays (µHM) engineered from human induced pluripotent stem cells (iPSCs). Elongated tissue geometries elicited cardiomyocyte shape and electrophysiology changes led to adaptations that yielded increased calcium intake during each contraction cycle. Strikingly, pharmacologic studies revealed that a threshold of prestress and/or cellular alignment is required for sodium channel function, whereas L-type calcium and rapidly rectifying potassium channels were largely insensitive to these changes. Concurrently, tissue elongation upregulated sodium channel (NaV1.5) and gap junction (Connexin 43, Cx43) protein expression. Based on these observations, we leveraged elongated µHM to study the impact of loss-of-function mutation in Plakophilin 2 (PKP2), a desmosome protein implicated in arrhythmogenic disease. Within µHM, PKP2 knockout cardiomyocytes had cellular morphology similar to what was observed in isogenic controls. However, PKP2-/- tissues exhibited lower conduction velocity and no functional sodium current. PKP2 knockout µHM exhibited geometrically linked upregulation of sodium channel but not Cx43, suggesting that post-translational mechanisms, including a lack of ion channel-gap junction communication, may underlie the lower conduction velocity observed in tissues harboring this genetic defect. Altogether, these observations demonstrate that simple, scalable micro-tissue systems can provide the physiologic stresses necessary to induce electrical remodeling of iPS-CM to enable studies on the electrophysiologic consequences of disease-associated genomic variants.

Dynamic control of contractile resistance to iPSC-derived micro-heart muscle arrays.

Many types of cardiovascular disease are linked to the mechanical forces placed on the heart. However, our understanding of how mechanical forces exactly affect the cellular biology of the heart remains incomplete. In vitro models based on cardiomyocytes derived from human induced pluripotent stem cells (iPSC-CM) enable researchers to develop medium to high-throughput systems to study cardiac mechanobiology at the cellular level. Previous models have been developed to enable the study of mechanical forces, such as cardiac afterload. However, most of these models require exogenous extracellular matrix (ECM) to form cardiac tissues. Recently, a system was developed to simulate changes in afterload by grafting ECM-free micro-heart muscle arrays to elastomeric substrates of discrete stiffnesses. In the present study, we extended this system by combining the elastomer-grafted tissue arrays with a magnetorheological elastomeric substrate. This system allows iPSC-CM based micro-heart muscle arrays to experience dynamic changes in contractile resistance to mimic dynamically altered afterload. Acute changes in substrate stiffness led to acute changes in the calcium dynamics and contractile forces, illustrating the system's ability to dynamically elicit changes in tissue mechanics by dynamically changing contractile resistance.

Amide-Directed, Rhodium-Catalyzed Regio- and Enantioselective Hydroacylation of Internal Alkenes with Unfunctionalized Aldehydes.

Despite the current progress achieved in asymmetric hydroacylations, highly enantioselective catalytic addition of unfunctionalized aldehydes to internal alkenes remains an unsolved challenge. Here, using a coordination-assisted strategy, we developed a rhodium-catalyzed regio- and enantioselective addition of unfunctionalized aldehydes to internal alkenes such as enamides and ß,γ-unsaturated amides. Valuable α-amino ketones and 1,4-dicarbonyl compounds were directly obtained with high enantioselectivity from readily available materials.

Instance Segmentation and Ensemble Learning for Automatic Temperature Detection in Multiparous Sows.

The core body temperature serves as a pivotal physiological metric indicative of sow health, with rectal thermometry prevailing as a prevalent method for estimating core body temperature within sow farms. Nonetheless, employing contact thermometers for rectal temperature measurement proves to be time-intensive, labor-demanding, and hygienically suboptimal. Addressing the issues of minimal automation and temperature measurement accuracy in sow temperature monitoring, this study introduces an automatic temperature monitoring method for sows, utilizing a segmentation network amalgamating YOLOv5s and DeepLabv3+, complemented by an adaptive genetic algorithm-random forest (AGA-RF) regression algorithm. In developing the sow vulva segmenter, YOLOv5s was synergized with DeepLabv3+, and the CBAM attention mechanism and MobileNetv2 network were incorporated to ensure precise localization and expedited segmentation of the vulva region. Within the temperature prediction module, an optimized regression algorithm derived from the random forest algorithm facilitated the construction of a temperature inversion model, predicated upon environmental parameters and vulva temperature, for the rectal temperature prediction in sows. Testing revealed that vulvar segmentation IoU was 91.50%, while the predicted MSE, MAE, and R2 for rectal temperature were 0.114 °C, 0.191 °C, and 0.845, respectively. The automatic sow temperature monitoring method proposed herein demonstrates substantial reliability and practicality, facilitating an autonomous sow temperature monitoring.

CAT-CBAM-Net: An Automatic Scoring Method for Sow Body Condition Based on CNN and Transformer.

Sow body condition scoring has been confirmed as a vital procedure in sow management. A timely and accurate assessment of the body condition of a sow is conducive to determining nutritional supply, and it takes on critical significance in enhancing sow reproductive performance. Manual sow body condition scoring methods have been extensively employed in large-scale sow farms, which are time-consuming and labor-intensive. To address the above-mentioned problem, a dual neural network-based automatic scoring method was developed in this study for sow body condition. The developed method aims to enhance the ability to capture local features and global information in sow images by combining CNN and transformer networks. Moreover, it introduces a CBAM module to help the network pay more attention to crucial feature channels while suppressing attention to irrelevant channels. To tackle the problem of imbalanced categories and mislabeling of body condition data, the original loss function was substituted with the optimized focal loss function. As indicated by the model test, the sow body condition classification achieved an average precision of 91.06%, the average recall rate was 91.58%, and the average F1 score reached 91.31%. The comprehensive comparative experimental results suggested that the proposed method yielded optimal performance on this dataset. The method developed in this study is capable of achieving automatic scoring of sow body condition, and it shows broad and promising applications.

The Photocatalytic Performance of P, Cl Doped Carboxylated Multiwalled Carbon Nanotube Modified Graphitic Carbon Nitride.

Graphitized carbonitride (g-C3N4) is widely used in CO2 reduction, hydrogen production, and degradation of toxic chemical dyes and antibiotics. It is a kind of photocatalytic material with excellent performance, and it has the advantages of being safe and nontoxic, having a suitable band gap (2.7 eV), and having a simple preparation and high stability, but because of its fast optical recombination speed and low visible light overutilization, the multifunctional application of g-C3N4 is seriously hindered. Compared with pure g-C3N4, MWCNTs/g-C3N4 have a red-shift in the visible range and a strong absorption in the visible region. Melamine and carboxylated multiwalled carbon nanotubes were used as raw materials to successfully prepare CMWCNT modified g-C3N4 doped with P, Cl by a high temperature calcination method. The effect of the addition amount of P, Cl on the photocatalytic performance of modified g-C3N4 was studied. The experimental results show that the multiwalled carbon nanotubes can accelerate the electron migration, and the doping of P, Cl elements can change the energy band structure of g-C3N4 and reduce the band gap. Through fluorescence analysis and photocurrent analysis, it is known that the incorporation of P, Cl reduces the recombination efficiency of photogenerated electron-hole pairs. In order to explore the application in the degradation of chemical dyes, the photocatalytic degradation efficiency of RhB under visible light was studied. The photocatalytic performance of the samples was evaluated by photodecomposition of aquatic hydrogen. The results showed that when the amount of ammonium dihydrogen phosphate was 10 wt %, the photocatalytic degradation efficiency was the highest, which was 21.13 times higher than that of g-C3N4.

Schizophyllum commune Reduces Expression of the SARS-CoV-2 Receptors ACE2 and TMPRSS2.

The current global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of COVID-19 has infected hundreds of millions of people, killed millions, and continues to pose a threat. It has become one of the largest epidemics in human history, causing enormous damage to people's lives and economies in the whole world. However, there are still many uncertainties and continued attention to the impact of SARS-CoV-2 on human health. The entry of SARS-CoV-2 into host cells is facilitated by the binding of the spike protein on the virus surface to the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Furthermore, transmembrane protease serine 2 (TMPRSS2) is a host surface protease that cleaves and proteolytically activates its S protein, which is necessary for viral infection. Thus, SARS-CoV-2 uses the ACE2 receptor for cell entry and initiates the S protein using the protease TMPRSS2. Schizophyllum commune (SC) is one of the most widely distributed fungi, often found on the rotten wood of trees that has been found to have various health benefits, including anticancer, antimicrobial activity, antiparasitic, and immunomodulatory function. In this article, SC significantly diminished the expression ACE2 and TMPRSS2 protein in vitro and in vivo without cell damage. In addition, adenosine from SC was also proven in this experiment to reduce the ACE2 and TMPRSS2 expression. Thus, our findings suggest that SC and adenosine exhibit potential for the repression of SARS-CoV-2 infection via the ACE2 and TMPRSS2 axis.

Scientific Evidences of Calorie Restriction and Intermittent Fasting for Neuroprotection in Traumatic Brain Injury Animal Models: A Review of the Literature.

It has widely been accepted that food restriction (FR) without malnutrition has multiple health benefits. Various calorie restriction (CR) and intermittent fasting (IF) regimens have recently been reported to exert neuroprotective effects in traumatic brain injury (TBI) through variable mechanisms. However, the evidence connecting CR or IF to neuroprotection in TBI as well as current issues remaining in this research field have yet to be reviewed in literature. The objective of our review was therefore to weigh the evidence that suggests the connection between CR/IF with recovery promotion following TBI. Medline, Google Scholar and Web of Science were searched from inception to 25 February 2022. An overwhelming number of results generated suggest that several types of CR/IF play a promising role in promoting post-TBI recovery. This recovery is believed to be achieved by alleviating mitochondrial dysfunction, promoting hippocampal neurogenesis, inhibiting glial cell responses, shaping neural cell plasticity, as well as targeting apoptosis and autophagy. Further, we represent our views on the current issues and provide thoughts on the future direction of this research field.

Intermittent fasting enhances hippocampal NPY expression to promote neurogenesis after traumatic brain injury.

OBJECTIVES: Interventions for preventing cognitive dysfunction after traumatic brain injury (TBI) are limited. Given that adult hippocampal neurogenesis after brain injury contributes to cognitive recovery, and hippocampal neurogenesis is potentially affected by nutritional factors, the aim of this study was to examine whether fasting could promote hippocampal neurogenesis and thus ameliorate the cognitive defects after TBI. METHODS: The present study used 8- to 10-wk-old C57 BL/6 N mice weighing 23 g, half males and half females. The mice were randomly assigned to each group, with 10 to 18 mice per group. All mice were housed in an approved animal facility with a 12-h light/dark cycle. In the metabolic study (food intake, body weight, blood glucose, triacylglycerol, total cholesterol, and ß-hydroxybutyric acid ), 54 mice (male:female = 1:1) were randomized to the ad libitum (AL) group (n = 18) and the intermittent fasting (IF) group (n = 36). In the neurogenesis study, 45 mice (male:female = 1:1) were randomized to AL (n  = 18), IF (n  = 9), IF + scramble (n  = 9), and the IF + neuropeptide Y (NPY)_siRNA (n  = 9) groups. In the Morris water maze test, 48 mice (male:female = 1:1) were randomized to AL (n  = 12), IF (n  = 12), IF + scramble (n  = 12), and the IF + NPY_siRNA (n  = 12) groups. RESULTS: We showed that a 1-mo-long IF regimen enhanced the proliferation of neural stem cells in the subgranular zone of the hippocampus 3 d after TBI, in addition to improving the cognitive performance in the Morris water maze test. Furthermore, an increase in the hippocampal NPY expression was detected in the IF group after the injury, compared with the mice fed AL, and local knockdown of NPY in vivo attenuated the effects of IF on TBI. CONCLUSIONS: These findings suggest that IF promotes hippocampal neurogenesis after TBI by a mechanism that involves enhancement of NPY expression, to alleviate cognitive dysfunction caused by injury.

Optogenetics for Understanding and Treating Brain Injury: Advances in the Field and Future Prospects.

Optogenetics is emerging as an ideal method for controlling cellular activity. It overcomes some notable shortcomings of conventional methods in the elucidation of neural circuits, promotion of neuroregeneration, prevention of cell death and treatment of neurological disorders, although it is not without its own limitations. In this review, we narratively review the latest research on the improvement and existing challenges of optogenetics, with a particular focus on the field of brain injury, aiming at advancing optogenetics in the study of brain injury and collating the issues that remain. Finally, we review the most current examples of research, applying photostimulation in clinical treatment, and we explore the future prospects of these technologies.

Determination of Phenolic Compounds in Beverages by Three-Flow Channel Isocratic HPLC with Electrochemical Detections Using a Column-Switching Technique.

For the quantitative analysis of phenolic compounds in beverage samples, a three-flow channel isocratic HPLC with electrochemical detection (3LC-ECD) system was devised using a column-switching technique. Phenolic compounds with significantly different hydrophobicity (the range of calculated log P: -0.77 to 3.02) were simultaneously measured to draw three chromatograms by the 3LC-ECD; the peaks of gallic acid (GA), protocatechuic acid (PCA), and gallocatechin (GC) were observed at electrochemical detector 1 (D1) within 42 min, the peaks of procyanidin B3 (B3), epigallocatechin (EGC), catechin (C), epicatechin (EC), procyanidin B2 (B2), ethyl gallate (Eg), and epigallocatechin gallate (EGCg) were observed at D2 within 50 min, and the peaks of epicatechin gallate (ECg), gallocatechin gallate (GCg), catechin gallate (Cg), propyl gallate (Pg), and resveratrol (RVT) were observed at D3 within 70 min. The relationships between the phenolic compound concentrations and their chromatographic peak heights gave good linearity with correlation coefficients of more than 0.998. The detection limits of the phenolic compounds by the 3LC-ECD ranged from 0.6 to 3.0 µg/L. Further, the phenolic compound concentrations of commercially available teas and wines were determined with a relative standard deviation (RSD) of less than 4.9% (n = 6), and their recoveries ranged from 91 to 109%. These results indicate that the 3LC-ECD system provided an accurate, precise, and specific determination of the phenolic compounds in beverages without affecting the matrices derived from these samples.

Regio- and Enantioselective (3+3) Cycloaddition of Nitrones with 2-Indolylmethanols Enabled by Cooperative Organocatalysis.

The regio- and enantioselective (3+3) cycloaddition of nitrones with 2-indolylmethanols was accomplished by the cooperative catalysis of hexafluoroisopropanol (HFIP) and chiral phosphoric acid (CPA). Using this approach, a series of indole-fused six-membered heterocycles were synthesized in high yields (up to 98 %), with excellent enantioselectivities (up to 96 % ee) and exclusive regiospecificity. This approach enabled not only the first organocatalytic asymmetric (3+3) cycloaddition of nitrones but also the first C3-nucleophilic asymmetric (3+3) cycloaddition of 2-indolylmethanols. More importantly, theoretical calculations elucidated the role of the cocatalyst HFIP in helping CPA control the reactivity and enantioselectivity of the reaction, demonstrating a new mode of cooperative catalysis.

A comparative study of sociality between firstborn children and only children of young age / 中国学校卫生

Objective@#To understand the current situation of the social development between only children and firstborn of young age, so as to provide a reference for the promotion of the social development of young children.@*Methods@#Stratified cluster sampling method was used to select 734 only children and firstborn children aged 3-9 in two kindergartens and two primary schools from grade 1 to grade 3 for questionnaire survey in Bengbu City. The content included the general information of children and their parents and the social assessment of children.@*Results@#The rate of emotional symptoms in firstborn children(27.8%) was higher than those of only children (17.6%)( χ 2=9.45, P <0.01). The results of univariate analysis showed that the rate of hyperactivity and inattention in social development of both only children and firstborns decreased with the increase of family socioeconomic status ( P < 0.05 ). Multivariate Logistic regression analysis of only children showed that only children with high economic status had a lower risk of hyperactivity and inattention and had a higher risk of peer interaction( P <0.05). The prosocial behavior of girls was better than that of boys in the aspect of social development of only children and firstborn children( OR =1.70, 2.85, P <0.05). For only children, the occurrence risk of being difficult was lower when the primary caregiver was parents than grandparents( OR =1.63, P < 0.05 ). For firstborn children, the risk of being difficult in nuclear families was lower than that in third generation families( OR = 2.14 , P <0.05). Multivariate analysis of the only child showed that boys had higher risk of hyperactive attention and less prosocial behavior than girls ( OR =2.24, 1.70, P <0.05), and a lower risk of developing mood disorders than girls( OR =0.57, P <0.05). The social development of only children varied among different grades, and the risk of abnormal prosocial behavior was lower with the increase of grades ( P <0.05).@*Conclusion@#Higer family social status is positively associated with children s social development level. But parents with high economic status should also avoid too much material and spiritual doting. Parents should strengthen their own learning to enhance the level of socialized education, raising siblings equally, improve the quality of parent child relationship, and promote the all round development of children s socialization level.

A New Triangular Rotation and Advancement Pulp Flap for Lateral Oblique Fingertip Defect.

The fingertip is one of the most common sites of traumatic injuries faced by hand surgeons. In cases of lateral oblique amputation, only limited alternatives are available for reconstruction. This study introduced a new method involving rotation and use of an advancement pulp flap for covering lateral oblique defect and evaluated its outcome. METHOD: A series of 12 patients with 14 lateral oblique fingertip defects were recruited in this study. All fingertips were unreplantable and were injured distal to the proximal one-third of nail bed, with phalanx exposed. All cases received surgical reconstruction using a triangular rotation and advancement pulp flap. Static 2-point discrimination, cold intolerance, pain, hypersensitivity, range of motion, and aesthetic satisfaction were evaluated 6 months to 12 months postoperation. RESULT: Bone defect was noted in 7 cases. The area of defect was 10×7-20×12 mm2, and the angle of defect was 30-60 degrees. Mean follow-up was 14.3 months. No hook nail deformity, cold intolerance, and hypersensitivity were observed. One patient complained about pain postoperation, demanding a second operation. Static 2-point discrimination was between 5 and 8 mm in all cases. Range of motion of distal interphalangeal joint recovered to 20-45 degrees at the last follow-up. No stiffness was observed in the interphalangeal or metacarpophalangeal joints. All patients were satisfied with the appearance of the flap. CONCLUSION: The triangular rotation and advancement pulp flap is simple, safe, and reliable for treating lateral oblique defect of fingertip, providing scope for anatomical reconstruction and fair sensation and aesthetic recovery.

Impact of father’s parenting effectiveness and children’s sociality on the acceptance of "second-born children" / 中国学校卫生

Objective@#To explore the interrelationships among fathers’ parenting effectiveness, urban children’s sociality and the acceptance of “second-born children” by children (including only children and “first-born children”), and to provide a reference for interventional measured for improving children’s acceptance rate of "second-bron children".@*Methods@#Stratified cluster sampling method was used to select 3 to 9-year-old 778 only children and “first-born children” in two kindergartens and grades 1-3 of two primary schools in a city, and the children and their fathers were investigated with questionnaires survey regarding general information of both fathers and children, the acceptance degree of “second-born children”, the sociality of children and the parenting effectiveness of fathers.@*Results@#Children in different grades had different acceptance levels of “second-born children” and the difference was statistically significant (F=2.89，P<0.05). Correlation analysis showed that the acceptance of “second-born children” was negatively correlated with the interaction between the only children and “first-born children”, and positively correlated with the prosocial behavior of the only children and “first-born children”, both of which are statistically significant (r=-0.10，0.10,P<0.01). Father’s role efficacy was negatively correlated with onlychildren and “first-born-children” peer interaction problems(r=-0.15), and positively correlated with prosocial behavior(r=0.32), both of which were statistically significant (P<0.01). The structural equation model of father’s parenting effectiveness, children’s sociality and acceptance of “second-born children” showed that low father’s parenting effectiveness could positively predict children’s peer communication problems and negatively predict children’s prosocial behavior, and thus children’s peer communication problems could negatively predict the acceptance degree of only children and “first-born children” to “second-born children”, and prosocial behaviors could positively predict the acceptance degree of “second-born children”.@*Conclusion@#Promoting father’s parenting efficiency can facilitate the social development of children and the acceptance of siblings.It’s recommended that fathers actively participate in specific parenting matters for the only child and "one child",and consciously improve parenting effectiveness.

A Preliminary Report of the Relationship Between Gene Polymorphism of IL-8 and Its Receptors and Systemic Inflammatory Response Syndrome Caused by Wasp Stings.

The plasma levels of interleukin-8 (IL-8) and its receptors (CXCR1 and CXCR2) play a significant role in the development of systemic inflammatory response syndrome (SIRS), but it is not clear how these proteins are involved in wasp sting patients developing SIRS. To study potential genetic factors predisposing to the risk of SIRS caused by wasp sting injury, we determined the plasma levels of IL-8 and its receptors among SIRS patients with wasp sting injury and investigated the association of single-nucleotide polymorphisms of these genes with SIRS. A total of 225 patients were divided into the SIRS group (n = 62) and non-SIRS group (control, n = 163), and we associated polymorphisms in IL-8 [rs4073 (-251T>A), rs2227532 (-845C>T), rs2227307 (+396G>T), rs2227306 (+781T>C), CXCR1 rs2234671 (+860C>G), CXCR2 [rs2230054 (+811T>C), rs57929613 (+1235C>T), and rs60626131 (+1440A>G)] with SIRS with a linear additive model. In terms of protein expression, the IL-8, CXCR1, and CXCR2 plasma levels were significantly higher in the SIRS group than in the control group (p < 0.001). Significantly higher frequencies were observed for the IL-8 - 251T allele (AT+TT), CXCR2 + 811T allele (CT+TT), and +1235C allele (TC+CC) in the SIRS group, when compared with the control group, with odds ratio (OR) = 3.971 (95% confidence interval [CI], 1.618-9.734), p = 0.003; OR = 4.223 (95% CI, 1.863-9.571), p = 0.001; and OR = 4.012 (95% CI, 1.773-9.079), p = 0.001; respectively. In addition, SIRS is more likely to occur in males, patients with number of wasp stings ≥10 stings, and stings in the limbs. The current study suggests that the IL-8 - 251T allele (AT+TT) and IL-8 receptor CXCR2 + 811C allele (CT+TT) and +1235T allele (TC+CC) could be risk factors among SIRS patients with wasp sting injury.

Robust Superhydrophobic Conical Pillars from Syringe Needle Shape to Straight Conical Pillar Shape for Droplet Pancake Bouncing.

Superhydrophobic conical pillars have great industrial application potential in, for example, anti-icing of aircraft wings and protecting high voltage transmission lines from freezing rain because of their droplet pancake bouncing phenomenon, which is recognized to further reduce the liquid-solid contact time. However, there are still no methods that can fabricate robust superhydrophobic conical pillars in large scale. Here, a mold replication technology was proposed to realize the large-scale fabrication of superhydrophobic conical pillars with high mechanical strength. An Al mold with intensive conical holes decorated with micro/nanometer-scale structures was fabricated by nanosecond laser drilling and HCl etching. The conical shape originated from a near Gaussian spatial distribution of the energy and temperature in the radial direction in the laser drilling processes. Robust superhydrophobic conical pillars from syringe needle shape to straight conical pillar shape were easily fabricated through replication from the Al mold without any extra spray of superhydrophobic nanoparticles. It was also found that although all superhydrophobic conical pillars with different shapes could generate the droplet pancake bouncing, the shape had a great influence on the critical bottom space and the critical Weber number (We) to generate pancake bouncing. The pancake bouncing with the shortest contact time of a 68.5% reduction appeared on superhydrophobic straight conical pillars with the shape angle of 180°. Overcoming the difficulties in the large-scale fabrication and robustness of superhydrophobic conical pillars will promote practical applications of the droplet pancake bouncing phenomenon.

Genomewide DNA Methylation Responses in Patients with ß-Thalassemia Treated with Yisui Shengxue Granules ().

OBJECTIVE: To examine the clinical effects of Yisui Shengxue Granules () in the treatment of ß-thalassemia and explore its mechanism on DNA methylation levels. METHODS: A randomized placebo-controlled double-blinded trial was conducted. Forty patients with ß-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group (12 g/bag three times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin (Hb), red blood cells (RBCs), reticulocytes (Ret), and fetal hemoglobin (HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0. RESULTS: Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group (P<0.01 or P<0.05). Meanwhile, HbF increased from the 2nd to the 3rd month (P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor (IGF1R) and Janus kinase 3 (JAK3) revealed the most relations with other genes (degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2 (PLCG2) and mitogen-activated protein kinase 10 (MAPK10) showed a stronger intermediary role (betweenness centrality=0.04). CONCLUSIONS: JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis. (Registration No. NCT01549080).

Biopanning of allergens from wasp sting patients.

OBJECTIVE: Wasp venom is a potentially important natural drug, but it can cause hypersensitivity reactions. The purpose of the present study was to systematically study the epitopes of wasp venom. METHODS: Using a random 12-peptide phage library, we performed antibody-binding epitope panning on ten serum samples from wasp sting victims at 3 h and 4 days after the sting. The panning epitopes were identified by high-throughput sequencing and matched with wasp venom proteins by BLAST. The panned antibody-binding epitopes were verified by ELISA. RESULTS: A total of 35 specific potential wasp venom epitopes in 4 days were identified. Amongst them, twelve peptide epitopes were matched with nine wasp venom proteins, namely, vitellogenin precursor, hexamerin 70b precursor, venom carboxylesterase-6 precursor, MRJP5, major royal jelly protein 8 precursor, venom acid phosphatase Acph-1 precursor, phospholipase A2, venom serine protease 34 precursor, and major royal jelly protein 9 precursor. The changes in serum IgM antibodies induced by wasp venom were confirmed by ELISA based on the 12 peptide epitopes. CONCLUSION: The nine wasp venom proteins are potential allergens, which should be excluded or modified in the potential biomedical applications of wasp venom.