Wuzi Yanzong Pill Plays A Neuroprotective Role in Parkinson's Disease Mice via Regulating Unfolded Protein Response Mediated by Endoplasmic Reticulum Stress / 中国结合医学杂志

OBJECTIVE@#To investigate the protective effects and its possible mechanism of Wuzi Yanzong Pill (WYP) on Parkinson's disease (PD) model mice.@*METHODS@#Thirty-six C57BL/6 male mice were randomly assigned to 3 groups including normal, PD, and PD+WYP groups, 12 mice in each group. One week of intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to establish the classical PD model in mice. Meanwhile, mice in the PD+WYP group were administrated with 16 g/kg WYP, twice daily by gavage. After 14 days of administration, gait test, open field test and pole test were measured to evaluate the movement function. Tyrosine hydroxylase (TH) neurons in substantia nigra of midbrain and binding immunoglobulin heavy chain protein (GRP78) in striatum and cortex were observed by immunohistochemistry. The levels of TH, GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1α, XBP1, ATF6, CHOP, ASK1, p-JNK, Caspase-12, -9 and -3 in brain were detected by Western blot.@*RESULTS@#Compared with the PD group, WYP treatment ameliorated gait balance ability in PD mice (P<0.05). Similarly, WYP increased the total distance and average speed (P<0.05 or P<0.01), reduced rest time and pole time (P<0.05). Moreover, WYP significantly increased TH positive cells (P<0.01). Immunofluorescence showed WYP attenuated the levels of GRP78 in striatum and cortex. Meanwhile, WYP treatment significantly decreased the protein expressions of GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1 α, XBP1, CHOP, Caspase-12 and Caspase-9 (P<0.05 or P<0.01).@*CONCLUSIONS@#WYP ameliorated motor symptoms and pathological lesion of PD mice, which may be related to the regulation of unfolded protein response-mediated signaling pathway and inhibiting the endoplasmic reticulum stress-mediated neuronal apoptosis pathway.

Effect of functional training on adolescent health / Efeito do treinamento funcional na saúde do adolescente / Efecto del entrenamiento funcional en la salud del adolescente

ABSTRACT Introduction The high rate of obesity caused by the modern sedentary lifestyle associated with bad eating habits and the high intellectual demand at school generates a physical and psychological deficit in contemporary youth. The damage caused in this growth phase is irrecoverable, and the habits acquired during this period are crucial to individual health. The urgency of this problem has generated research on methods to reverse this situation, among them functional training. Objective To study the effect of functional training on adolescent health. Methods 30 boys and 30 girls were recruited as study subjects. The experiment lasted one semester, including two weekly physical education classes focused on functional training. Data on body indexes, fitness, and physical test results were collected before and after the intervention. The results of the experiment were integrated and compared. Results The selection of functional training can optimize adolescents' biochemical indexes, making adolescents' orthostatic posture more balanced, and significantly improve muscle strength, body flexibility, explosive strength of upper and lower extremities, motor coordination, body flexibility, muscular endurance, cardiopulmonary function, etc. Conclusion Functional training can effectively improve the physical health of adolescents. Participants have increased their interest in sports, and this intervention can potentially promote the development of adolescents' physical and mental health if replicated nationwide. Studies are needed to popularize functional training in adolescents. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução O elevado índice de obesidade ocasionada pelo sedentarismo moderno associado a maus hábitos alimentares e a alta demanda intelectual escolar geram um déficit físico e psicológico na juventude contemporânea. Os danos ocasionados nessa fase de crescimento são irrecuperáveis e os hábitos adquiridos nesse período são determinantes para a saúde individual. A urgência desse problema tem gerado pesquisas sobre métodos para reverter essa situação, dentre eles desponta o treinamento funcional. Objetivo Estudar o efeito do treinamento funcional na saúde do adolescente. Métodos 30 meninos e 30 meninas foram recrutados como sujeitos de estudo. O experimento durou um semestre, incluindo duas aulas semanais de educação física, com foco no treinamento funcional. Os dados dos índices corporais, aptidão física e resultados dos testes físicos foram coletados antes e depois da intervenção. Os resultados do experimento foram integrados e comparados. Resultados A seleção do treinamento funcional pode otimizar os índices bioquímicos dos adolescentes, tornar a postura ortostática dos adolescentes mais equilibrada, melhorar significativamente a força muscular, a flexibilidade corporal, a força explosiva das extremidades superior e inferior coordenação motora, flexibilidade corporal, resistência muscular, função cardiopulmonar, etc. Conclusão O treinamento funcional pode melhorar efetivamente a saúde física dos adolescentes. Os participantes aumentaram seu interesse pelo esporte e essa intervenção tem o potencial de promover o desenvolvimento da saúde física e mental dos adolescentes caso replicada a nível nacional. Estudos são necessários para a popularização do treinamento funcional em adolescentes. Nível de evidência II; Estudos terapêuticos - investigação dos desfechos do tratamento.

RESUMEN Introducción La alta tasa de obesidad causada por el sedentarismo moderno asociado a malos hábitos alimenticios y la alta exigencia intelectual escolar generan un déficit físico y psicológico en la juventud contemporánea. Los daños causados en esta fase de crecimiento son irrecuperables y los hábitos adquiridos durante este periodo son cruciales para la salud del individuo. La urgencia de este problema ha generado la investigación de métodos para revertir esta situación, entre ellos surge el entrenamiento funcional. Objetivo Estudiar el efecto del entrenamiento funcional en la salud de los adolescentes. Métodos Se reclutaron 30 chicos y 30 chicas como sujetos de estudio. El experimento duró un semestre, incluyendo dos clases semanales de educación física, centradas en el entrenamiento funcional. Se recogieron datos sobre los índices corporales, la forma física y los resultados de las pruebas físicas antes y después de la intervención. Los resultados del experimento se integraron y compararon. Resultados La selección del entrenamiento funcional puede optimizar los índices bioquímicos de los adolescentes, hacer que la postura ortostática de los adolescentes sea más equilibrada, mejorar significativamente la fuerza muscular, la flexibilidad corporal, la fuerza explosiva de las extremidades superiores e inferiores la coordinación motora, la flexibilidad corporal, la resistencia muscular, la función cardiopulmonar, etc. Conclusión El entrenamiento funcional puede mejorar eficazmente la salud física de los adolescentes. Los participantes han aumentado su interés por el deporte y esta intervención tiene el potencial de promover el desarrollo de la salud física y mental de los adolescentes si se reproduce a nivel nacional. Se necesitan estudios para la popularización del entrenamiento funcional en adolescentes. Nivel de evidencia II; Estudios terapêuticos - investigación de los resultados del tratamiento.

Cloning the full-length CDNA of actin gene and analysing alliinase gene expression in tillering onion

Tillering onion is a herbaceous plant belonging to the Liliaceae family. We cloned the cDNAs of the actin gene (AcACT, GenBank: MF919598) of tillering onion using rapid amplification of the cDNA ends. The full-length cDNA of AcACT was 1,357 bp long with an open reading frame of 1,131 bp encoding 376 amino acids. The amino acid sequence of AcACT shared > 96% similarity with the amino acid sequences of other ACTs and was found (by means of phylogenetic tree analysis) to be closely related to those of Ananas comosus and Papaver somniferum. AcACT expressions showed no significant differences (p > 0.01) in two cultivars L-SH and L-SY over three growth periods and under suitable conditions, low temperature, and short-day conditions. In addition, AcACT was used as an internal reference gene to analyse the expression of the alliinase gene (AcALL). AcALL expression trends in the roots, stems and leaves were consistent with those of diallyl disulphide and diallyl trisulphide. Thus, AcACT is highly conserved and can be used as a suitable internal reference gene when analysing gene expression in tillering onion.

Acupoit is the "transducer" in the physiochemical information coupling response of acupuncture / 中国针灸

Acupoint is the initial response site of acupuncture stimulus and also the source link of the effect onset of acupuncture. Acupuncture is a mechanical physical stimulus. How is the mechanical force of acupuncture transduced into neuroelectrical and biochemical signals at acupoint? How does the physiochemical information of acupoint launch acupuncture effect? All of these remain the common and crucial questions in the study of acupuncture effect mechanism. Physical changes are induced in the local tissue of acupoint by needling techniques, such as the deformation and displacement of muscle fibers, which may act on the nerve ending receptors and produce electroneurographic signals. Besides, these changes may activate the mechanosensitive ion channels of the cytomembrane in acupoint site. Through cellular signal transduction, the physical signals may be transformed into chemical ones to trigger the physiochemical coupling response of acupoint microenvironment. Eventually, acupuncture effect is generated via nerves and body fluids. "The mechanical force of acupuncture", through "the physiochemical transduction", promotes the body's perception and transmits acupuncture signals. It suggests that acupoint is the "transducer" in the physiochemical information coupling response of acupuncture.

Effect of moxibustion on autophagy in mice with Alzheimer's disease based on mTOR/p70S6K signaling pathway / 中国针灸

OBJECTIVE@#To investigate the effect of moxibustion on autophagy and amyloid β-peptide1-42 (Aβ1-42) protein expression in amyloid precursor protein/presenilin 1 (APP/PS1) double-transgenic mice with Alzheimer's disease (AD).@*METHODS@#After 2-month adaptive feeding, fifty-six 6-month-old APP/PS1 double transgenic AD mice were randomly divided into a model group, a moxibustion group, a rapamycin group and an inhibitor group, 14 mice in each group. Another 14 C57BL/6J mice with the same age were used as a normal group. The mice in the moxibustion group were treated with monkshood cake-separated moxibustion at "Baihui"(GV 20), "Fengfu" (GV 16) and "Dazhui" (GV 14) for 20 min; the mice in the rapamycin group were intraperitoneally injected with rapamycin (2 mg/kg); the mice in the inhibitor group were treated with moxibustion and injection of 1.5 mg/kg 3-methyladenine (3-MA). All the treatments were given once a day for consecutive 2 weeks. The morphology of hippocampal tissue was observed by HE staining; the ultrastructure of hippocampal tissue was observed by transmission electron microscopy; the expression of Aβ1-42 protein in frontal cortex and hippocampal tissue was detected by immunohistochemistry; the expressions of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) protein in hippocampus were detected by Western blot method.@*RESULTS@#Compared with the normal group, the number of neuron cells was decreased, cells were necrotic and deformed, and autophagy vesicle and lysosome were decreased in the model group. Compared with the model group, the number of neuron cells was increased, cell necrosis was decreased, and autophagy vesicle and lysosome were increased in the moxibustion group and the rapamycin group. Compared with the normal group, the protein expressions of Aβ1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the model group were increased (P<0.05); compared with the model group, the protein expressions of Aβ1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group, rapamycin group and inhibitor group were decreased (P<0.05); compared with the inhibitor group, the protein expressions of Aβ1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group and rapamycin group were decreased (P<0.05); compared with the rapamycin group, the protein expressions of mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group were decreased (P<0.05).@*CONCLUSION@#Moxibustion could enhance autophagy in hippocampal tissue of APP/PS1 double transgenic AD mice and reduce abnormal Aβ aggregation in brain tissue, the mechanism may be related to the inhibition of mTOR/p70S6K signaling pathway.

Effect of Xingnao Kaiqiao acupuncture on safety of rt-PA intravenous thrombolysis in patients with cerebral infarction: a randomized controlled trial / 中国针灸

OBJECTIVE@#To assess the effect of Xingnao Kaiqiao (regaining consciousness and opening orifices) acupuncture on the efficacy and safety of intravenous thrombolysis with recombinant tissue plasminogen activator (rt-PA) in patients with cerebral infarction.@*METHODS@#A total number of 142 patients of cerebral infarction undergoing rt-PA intravenous thrombolysis were randomized into an acupuncture-medication group (71 cases) and a western medication group (71 cases, 1 case dropped off). In the western medication group, rt-PA intravenous thrombolysis was given. In the acupuncture-medication group, besides the intervention as the control group, Xingnao Kaiqiao acupuncture was provided at Shuigou (GV 26), Neiguan (PC 6), Sanyinjiao (SP 6), Jiquan (HT 1), etc. once daily. One treatment session contained 6 treatments and 1 session was required. Before and after treatment, the score of the National Institute of Health stroke scale (NIHSS), the levels of the relevant indexes of symptomatic intracerebral hemorrhage (sICH) (platelet [PLT], D-dimer and fibrinogen), the incidences of sICH and adverse effect were compared between groups. The efficacy was assessed in two groups.@*RESULTS@#After treatment, NIHSS scores and the levels of D-dimer were reduced compared with those before treatment in both groups (P<0.05), and those in the acupuncture-medication group were lower than the western medication group (P<0.05). The level of fibrinogen in the acupuncture-medication group was increased in comparison with that before treatment (P<0.05), and also higher than the western medication group (P<0.05). The incidence of sICH was 0% (0/71) in the acupuncture-medication group, lower than 8.6% (6/70) in the western medication group (P<0.05). The effective rate was 97.2% (69/71) in the acupuncture-medication group, higher than 87.1% (61/70) in the western medication group (P<0.05). The incidence of adverse effect was 2.8% (2/71) in the acupuncture-medication group, lower than 12.9% (9/70) in the western medication group (P<0.05).@*CONCLUSION@#Xingnao Kaiqiao acupuncture may improve the efficacy of rt-PA intravenous thrombolysis in the patients with cerebral infraction and decrease the incidences of sICH and adverse effect. The mechanism may be related to the regulation of fibrinogen and D-dimer levels.

Introduction of legislation of disciplinary system for acupuncturists in various states in the United States: taking Ohio and other 11 states as examples / 中国针灸

With 12 representative states such as Ohio and Illinois as examples, the disciplinary system for acupuncturists in the United States is introduced. The disciplinary system mainly covers several aspects such as the subject of the implementation, the types of illegal acts, the form of disciplinary responsibility, and the hearing procedure. The acupuncture industry association has the responsibility of disciplinary action in most states. Corresponding disciplinary measures will be taken depending on the illegal activities of acupuncturists according to the types of illegal practice specified by each state. The hearing procedure provides procedural guarantee for the subsequent rights protection activities of acupuncturists.

Stem Cell-Derived Exosomes: A New Method for Reversing Skin Aging

Senescence is an inevitable natural life process that involves structural and functional degeneration of tissues and organs. Recently, the process of skin aging has attracted much attention. Determining a means to delay or even reverse skin aging has become a research hotspot in medical cosmetology and anti-aging. Dysfunction in the epidermis and fibroblasts and changes in the composition and content of the extracellular matrix are common pathophysiological manifestations of skin aging. Reactive oxygen species and matrix metalloproteinases play essential roles in this process. Stem cells are pluripotent cells that possess self-replication abilities and can differentiate into multiple functional cells under certain conditions. These cells also possess a strong ability to facilitate tissue repair and regeneration. Stem cell transplantation has the potential for application in anti-aging therapy. Increasing studies have demonstrated that stem cells perform functions through paracrine processes, particularly those involving exosomes. Exosomes are nano-vesicular substances secreted by stem cells that participate in cell-to-cell communication by transporting their contents into target cells. In this chapter, the biological characteristics of exosomes were reviewed, including their effects on extracellular matrix formation, epidermal cell function, fibroblast function and antioxidation. Exosomes derived from stem cells may provide a new means to reverse skin aging.

Development and Application of Three-Dimensional Bioprinting Scaffold in the Repair of Spinal Cord Injury

Spinal cord injury (SCI) is a disabling and destructive central nervous system injury that has not yet been successfully treated at this stage. Three-dimensional (3D) bioprinting has become a promising method to produce more biologically complex microstructures, which fabricate living neural constructs with anatomically accurate complex geometries and spatial distributions of neural stem cells, and this is critical in the treatment of SCI. With the development of 3D printing technology and the deepening of research, neural tissue engineering research using different printing methods, bio-inks, and cells to repair SCI has achieved certain results. Although satisfactory results have not yet been achieved, they have provided novel ideas for the clinical treatment of SCI. Considering the potential impact of 3D bioprinting technology on neural studies, this review focuses on 3D bioprinting methods widely used in SCI neural tissue engineering, and the latest technological applications of bioprinting of nerve tissues for the repair of SCI are discussed. In addition to introducing the recent progress, this work also describes the existing limitations and highlights emerging possibilities and future prospects in this field.

Urinary Biomarkers for the Noninvasive Detection of Gastric Cancer

Gastric cancer (GC) is associated with high morbidity and mortality rates. Thus, early diagnosis is important to improve disease prognosis. Endoscopic assessment represents the most reliable imaging method for GC diagnosis; however, it is semi-invasive and costly and heavily depends on the skills of the endoscopist, which limit its clinical applicability.Therefore, the search for new sensitive biomarkers for the early detection of GC using noninvasive sampling collection methods has attracted much attention among scientists.Urine is considered an ideal biofluid, as it is readily accessible, less complex, and relatively stable than plasma and serum. Over the years, substantial progress has been made in screening for potential urinary biomarkers for GC. This review explores the possible applications and limitations of urinary biomarkers in GC detection and diagnosis.

Research progress of pharmacological effects and security of Yuebawei / 中国药房

Yuebawei refers to 8 kinds of genuine medicinal materials in south China area，namely Citri Grandis Exocarpium， Citrus reticulata， Amomum villosum， Pogostemon cablin， Morinda officinalis， Aquilaria sinensis， Citrus medica and Polygonum multiflorum. The results of this review show that， the pharmacological effects of Yuebawei are very extensive. It has significant effects in relieving cough and phlegm， anti-bacterial and anti-inflammatory， enhancing immunity， anti-oxidation， antidepressant， immune regulation and regulating gastrointestinal function. Its development and application prospects are broad， but most of the existing pharmacological studies stay on pharmacodynamic studies. In the aspect of the safety research of Yuebawei， there are few studies on the safety of other medicinal materials， except for the liver toxicity of P. multiflorum； at the same time， there are few comparative studies on the therapeutic advantages of Yuebawei and other local medicinal materials in clinical application. It is necessary to strengthen the research on the material basis of pharmacological action mechanism， and safety and efficacy observation of clinical medication for Yuebawei， so as to provide scientific basis for the development of new drugs and clinical promotion and application of Yuebawei.

Classification and reconstruction of bile duct in pediatric split liver transplantation / 器官移植

Objective To investigate the anatomical classification of left intrahepatic bile duct (LHD) and the pattern of bile duct reconstruction during pediatric split liver transplantation and their relationship with postoperative biliary complications. Methods Clinical data of 75 pediatric recipients undergoing split liver transplantation were analyzed retrospectively. Before splitting the donor liver, iopromide injection was used for retrograde cholangiography through the common bile duct. According to the patterns of intrahepatic bile ducts in the second, third and fourth segments, the anatomical classification of LHD of the donor liver was determined. The biliary reconstruction regimens for different classification types of LHD were summarized. The incidence and treatment of biliary complications after pediatric split liver transplantation were analyzed. Results Among 75 donor livers, the anatomical classification of LHD included 57 cases (76%) of type Ⅰ, 9 cases (12%) of type Ⅱ, 4 cases (5%) of type Ⅲ and 5 cases (7%) of type Ⅳ LHD, respectively. Among 75 pediatric recipients, 69 cases (53 cases of type Ⅰ, 8 type Ⅱ, 4 type Ⅲ and 4 type Ⅳ) underwent the left hepatic duct-jejunum Roux-en-Y anastomosis, 1 case received common bile duct-jejunum Roux-en-Y anastomosis (type Ⅳ), and 5 cases underwent the left hepatic duct-common bile duct end-to-end anastomosis (4 cases of type Ⅰ and 1 type Ⅱ). Postoperative biliary complications occurred in 6 cases (8%), including 3 cases of biliary anastomotic stenosis, 2 cases of biliary anastomotic leakage and 1 case of bile leakage on the hepatic resection surface. Among 6 recipients, 4 cases were classified as type Ⅰ and 2 cases of type Ⅲ LHD. No significant difference was observed in the incidence of biliary complications between typical type and anatomical variant type of LHD (all P > 0.05). Among 3 recipients with biliary anastomotic stenosis, 2 cases underwent percutaneous transhepatic cholangial and drainage (PTCD) and 1 case repeatedly received biliary-intestinal anastomosis. Two cases of biliary anastomotic leakage underwent PTCD and 1 case of bile leakage on the hepatic resection surface received local drainage. All 6 children survived after treatment. Conclusions Anatomical variation of LHD can be observed in 24% of donor livers, and type Ⅱ accounts for the highest proportion of 12%. Prior to donor liver splitting, routine cholangiography and fine biliary anastomosis may effectively lower the incidence of biliary complications. The incidence of postoperative biliary complications is not significantly associated with the anatomical classification of LHD.

Design and clinical application of simplified "All in one" hepatic vein reconstruction in right split liver transplantation / 器官移植

Objective To introduce the simplified "All in one" hepatic vein reconstruction in right split liver transplantation, and to investigate the clinical indications, surgical procedures and clinical prognosis of this technique. Methods Clinical data of 2 recipients undergoing right split liver transplantation were retrospectively analyzed, and the simplified "All in one" hepatic vein reconstruction of right liver lobe was summarized and analyzed. In 2 cases, the right liver lobe was split in vivo. In case 1, the liver parenchyma was split until the first and second porta hepatis, and then the liver was obtained by whole-liver perfusion after cutting off the right hepatic duct, and the hepatic blood vessels were isolated ex vivo. In case 2, the left liver lobe was obtained during splitting in vivo and the right liver lobe was obtained after perfusion in vivo. During donor liver splitting in two cases, the common trunk of the middle hepatic vein was maintained in the left liver lobe, the S5 and S8 hepatic veins of the right liver lobe were reconstructed by the same donor iliac artery, and directly anastomosed with the gap between the left and middle hepatic veins of the inferior vena cava, thus reconstructing the integrity of the posterior inferior vena cava. Results The simplified "All in one" right hepatic vein reconstruction was adopted. The anhepatic phase of two recipients undergoing liver transplantation was 41 and 36 min. After the liver was incised open, the blood flow of the donor liver was normal, the iliac artery bypass was fully filled, liver congestion or swelling was not observed, and liver function was properly recovered after surgery. Two recipients were subject to postoperative follow-up for 23 and 10 months, respectively, No complications related to hepatic venous outflow tracts were noted, such as hepatic vein and inferior vena cava. Conclusions "All in one" hepatic vein reconstruction may simplify the procedures of hepatic venous outflow tract reconstruction, shorten the anhepatic phase and reduce the incidence of postoperative hepatic vein complications in complete right liver split transplantation.

Expression and Clinical Significance of DNA Demethylase TET in the Tissues in Patients with Neuroblastoma / 中山大学学报(医学科学版)

ObjectiveTo detect the expression level of DNA demethylase ten eleven translocation TET1， TET2， TET3 in the tissues of neuroblastoma （NB） patients，and to explore the expression and clinical significance of TET in patients with neuroblastoma. MethodsImmunohistochemistry （IHC） was used to detect the expression levels of TET1， TET2 and TET3 in the tissues of 46 patients with neuroblastoma； clinical data from Target database were used to analyze the relationship between the expression of TET1， TET2， TET3 and prognosis of NB patients， and batch correlation analysis was conducted. SPSS 25.0 statistical software was used for analysis， and GraphPad Prism 8 was used for plotting. Results①Immunohistochemical results showed that the expression of TET3 in neuroblastoma patients with different levels of mitosis karyorrhexis index （MKI） was significantly different with statistical significance （P=0.037）. The expression level of TET3 was statistically different among different groups of tumor cell differentiation （P=0.041）， and high expression of TET3 was positively correlated with good prognosis of patients （P=0.020）. The expression of TET1 was positively correlated with the expression of β2-AR （r=0.347， P=0.023）. ②Bioinformatics analysis showed that the mRNA expression level of TET3 was positively correlated with the mRNA expression level of tumor suppressor genes such as activity-dependent neuroprotective protein （ADNP， a neuroprotective protein） and AT-rich interactive domain 2 （ARID2， a chromatin remodeling factor）， and negatively correlated with the mRNA expression level of oncogenes such as CD63 （fourth transmembrane protein） and Josephin domain-containing protein 2 （JOSD2， deubiquitination enzyme）. TARGET database showed that TET3 mRNA level was positively correlated with overall survival （OS） and event-free survival （EFS） of neuroblastoma patients （P=0.024， P=0.014）. ConclusionsThe expression of TET3 in NB patients has a certain relationship with the clinical features of neuroblastoma patients， and the high expression of TET3 is positively correlated with the good prognosis of NB patients， suggesting that TET3 may play a role as a tumor suppressive factor in neuroblastoma， and TET3 can be used as a potential indicator to predict the prognosis of NB patients. It has some value in the clinical evaluation of NB.

Prognostic and Immunological Role of ABHD5： a Pan-Cancer Analysis / 中山大学学报(医学科学版)

ObjectiveTo evaluate the ABHD5 expression in pan-cancer and its correlations with the main clinical stages， prognosis and immune cell infiltration. MethodsGTEx， TCGA， TIMER2.0， CPTAC databases， immunohistochemistry and western blot were used to analyze the expression levels of ABHD5 in different cancer tissues and adjacent tissues as well as correlations between ABHD5 expression and the main clinical stages. Kaplan-Meier Plotter， Oncolnc and R2 databases were used to explore the prognostic value of ABHD5. The relationship between ABHD5 and immune cell infiltration was analyzed by TCGA and TIMER2.0 databases. STRING and GEPIA databases were used to detect ABHD5-binding proteins and co-expression genes， which were then analyzed by GO and KEGG. ResultsThe mRNA and protein expression levels of ABHD5 were lower in cancer tissues than those in normal tissues in multiple cancer types， but higher in few cancer types. High-level expression of ABHD5 was related to better prognosis in 8 cancer types and related to worse prognosis in 2. ABHD5 expression levels were positively correlated with immune cell infiltration in 9 cancer types， and were the same with neutrophil infiltration and expression of immune checkpoints in pan-cancer. Enrichment analysis showed that ABHD5 was related to histone demethylation. ConclusionPossibly used as a potential prognostic predictor in pan-cancer， ABHD5 was also correlated with neutrophil infiltration， expression of immune checkpoints and histone demethylation.

Application of fingerprint combined with quantitative analysis of multi -components by single marker in quality evaluation of Xanthoceras sorbifolia / 中国药房

OBJECTIVE To establish the fingerprints of Xanthoceras sorbifolia and determine the contents of flavonoids . METHODS HPLC was adopted . Using epigallocatechin as reference ，the fingerprints of 11 batches（No. S1-S11）of X. sorbifolia were drawn with Similarity Evaluation System of Chromatographic Fingerprints of TCM （2004A edition ）. The similarity evaluation was conducted ，the common peaks were also confirmed . Cluster analysis （CA）and principal component analysis （PCA）were also performed. Epigallocatechin was selected as internal reference ，and quantitative analysis of multi -components by single marker （QAMS）was used to determine the contents of gallocatechin ，catechin，epicatechin，dihydromyricetin，taxifolin and myricetin in 16 batches（No. S1-S16）of X. sorbifolia. The results were compared with the results of one point external standard method and standard curve method . RESULTS There were 15 common peaks in 11 batches of X. sorbifolia，and the similarity of them were 0.910-1.000. A total of 7 common peaks were identified ，i.e. galliccatechin（peak 1），epigallocatechin（peak 2），catechin（peak 3），epicatechin（peak 5），dihydromyricetin（peak 6），taxifolin（peak 14）and myricetin （peak 15）. The results of CA showed that S5-S7 and S 9 were clustered into one category ，S8 and S 11 were clustered into one category ，S10 were clustered into one category，S1-S4 were clustered into one category . The results of PCA showed that accumulative variance contribution rate of 3 principal components was 99.24%；S5-S7 were clustered into one category ，S8-S11 were clustered into one category ，S3 and S 4 were clustered into one category ，S1 and S 2 were clustered into one category . With the exception of myricetin and a partial batches （S12，S14-S16） of catechin ，the RSDs measured by the three methods for galliccatechin ，catechin （remaining batches ）， epicatechin，dihydromyricetin and taxifolin in 16 batches of X. sorbifolia were less than 4% （n＝3）. CONCLUSIONS The established HPLC fingerprint and the method for content determination can be used for the quality control of X. sorbifolia. QAMS method can be used for the content determination of galliccatechin，epicatechin，dihydromyricetin and taxifolin .

Application of the small private online course-based blended teaching model in Human Parasitology teaching / 中国血吸虫病防治杂志

Small private online course (SPOC), an emerging network-based, student-centered blended teaching model, combines the advantages of online teaching and flipped classroom, aiming to effectively improve students’ autonomous learning ability and learning efficiency. Considering a remarkable decline in the course of Human Parasitology, we built an SPOC-based blended teaching model for Human Parasitology, covering pre-course learning, intra-course leaning, post-course consolidation and evaluation, and such a model was applied among grade 2019 students with specialty of clinical medicine. Following the application of the SPOC-based blended teaching model, most students had improvements in autonomous learning ability and activity, and understanding of human parasitology knowledge, and the examination score disparity was avoided.

Preparation and characterization of fenugreek leaf flavonoids and their protective effects against oxidative damage to hepatocytes / 中国中药杂志

The present study investigated the main components of fenugreek(Trigonella foenum-graecum L.) leaf flavonoids(FLFs) and their antioxidant activity. FLFs were prepared and enriched by solvent extraction, and the flavonoids were characterized by high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS). The protective effect of FLFs against H_2O_2-induced stress damage to L02 hepatocytes was also investigated. Firstly, the cell viability was measured by MTT assay. The oxidative stress injury model was induced by H_2O_2 in L02 cells. The release of lactate dehydrogenase(LDH), the content of reduced glutathione(GSH) and malondialdehyde(MDA), and the activities of superoxide dismutase(SOD) and catalase(CAT) were measured by assay kits. Hoechst fluorescence staining was performed to observe the cell apoptosis. The expression levels of c-Jun N-terminal kinase(JNK), extracellular signal-regulated kinase 1/2(ERK1/2), nuclear factor erythroid-2 related factor 2(Nrf2), heme oxygenase 1(HO-1), and their phosphorylated proteins were detected by Western blot. Based on the MS fragment ion information and data in databases, FLFs contained eight flavonoids with quercetin and kaempferol as the main aglycons. The cell viabi-lity assay revealed that as compared with the conditions in the H_2O_2 treatment group, 3.125-25 μg·mL~(-1) FLFs could increase the viability of L02 cells, reduce LDH release and MDA content in a dose-dependent manner, potentiate the activities of SOD, CAT, and GSH, decrease the phosphorylation of JNK and ERK1/2 proteins, and up-regulate the expression of Nrf2 and HO-1. The results of fluorescence staining showed that the nucleus of the H_2O_2 treatment group showed concentrated and dense strong blue fluorescence, while the blue fluorescence intensity of the FLFs group decreased significantly. FLFs showed a protective effect against H_2O_2-induced oxidative damage in L02 cells, and the underlying mechanism is associated with the enhancement of cell capability in clearing oxygen free radicals and the inhibition of apoptosis by the activation of the MAPKs/Nrf2/HO-1 signaling pathway. The antioxidant effect of fenugreek leaf is related to its rich flavonoids.

Optimization of formulation of paclitaxel nanosuspension encapsulated by erythrocyte membrane based on Box-Behnken method / 中国中药杂志

In view of the longevity and innate immune escape of red blood cells, this study designed the red blood cell membrane-coated paclitaxel nanosuspension [RBC-(PTX)NS] and investigated its physicochemical properties and antitumor effect in vitro. Paclitaxel nanosuspension [(PTX)NS] was prepared by ultrasonic precipitation and then RBC-(PTX)NS by ultrasonic coating. The formulation of(PTX)NS was optimized with Box-Behnken method and indexes of particle diameter, zeta potential, and stability. The morphology, particle diameter, stability, in vitro dissolution, and antitumor effect of(PTX)NS and RBC-(PTX)NS were characterized. The results showed that the particle diameter and zeta potential were(129.38±0.92) nm and(-22.41±0.48) mV, respectively, for the optimized(PTX)NS, while(142.5±0.68) nm and(-29.85±0.53) mV, respectively, for RBC-(PTX)NS. Under the transmission electron microscope,(PTX)NS was spherical and RBC-(PTX)NS had obvious core-shell structure. RBC-(PTX)NS remained stable for 5 days at 4 ℃. The in vitro dissolution test demonstrated that the cumulative release rate of RBC-(PTX)NS reached 79% within 20 min, which was significantly higher than that(25%) of(PTX)NS(P<0.05). As evidenced by MTT assay, RBC-(PTX)NS highly inhibited the proliferation of HepG2 cells in a dose-dependent manner. The cell membrane-coated nano-preparation preparation method is simple and reproducible. It improves the solubility of PTX and endows RBC-(PTX)NS with higher stability and stronger cytotoxicity. Thus, it is a new method for the delivery of PTX via nanocrystallization.

Construction of protoplast genetic transformation system for Mycena--symbiont of Gastrodia elata / 中国中药杂志

Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.