Designing a Trifoliate Flap for Isolated Congenital Alar Rim Defect in Pediatric Patients.

Isolated congenital alar rim defects are extremely rare, and there has been no standard technique for the reconstruction of remarkable aesthetic deformity. Herein, we introduce a trifoliate flap for the correction of isolated congenital alar rim defects in pediatric patients. Fifteen cases undergoing nasal alar sulcus rotation flap surgery were analyzed retrospectively. This rotation flap including 3 triangles was a modified flap based on prior studies. Clinical medical notes and photographs were reviewed. Patients' (or their parents) reported satisfactions with aesthetic outcome were also evaluated during the post-operative follow-up period. In all patients, the isolated congenital alar rim defects were successfully reconstructed. The rotation flap survived and the wound healed primarily. The follow-up period ranged from 6 to 22 months (average 11 months). There were no incidents of flap loss, step-off deformities, nasal obstruction, or alar retraction. At follow-up of post-operative 3 months, pale red scars were observed in the operative area in few patients (2/15). However, these scars gradually became invisible at post-operative 6 months. All patients (or their parents) were satisfied with the aesthetic outcome of this operation. This newly designed trifoliate flap can be an alternative method for the reconstruction of isolated congenital alar rim defects in pediatric patients. The scars of this procedure can be unobvious with fine surgical suture.

Contexte: Les anomalies congénitales isolées du pourtour de l'aile du nez sont extrêmement rares et il n'existe aucune technique de référence pour la reconstruction de cette difformité esthétique notable. Nous présentons ici un volet trifolié pour la correction des anomalies congénitales du pourtour de l'aile du nez chez des patients pédiatriques. Méthodes: Quinze cas de patients subissant une chirurgie avec rotation de lambeau de sillon de l'aile du nez ont été analysés rétrospectivement. Ce lambeau de rotation comportant trois triangles était une version modifiée d'un lambeau utilisé dans des études précédentes. Les notes médicales cliniques et les photographies ont été analysées. La satisfaction exprimée par les patients (ou leurs parents) à propos du résultat esthétique a été également évaluée au cours de la période de suivi postopératoire. Résultats: L'anomalie congénitale isolée du pourtour de l'aile du nez a été réparée avec succès chez tous les patients. Le lambeau de rotation a survécu et la plaie a guéri d'emblée: la durée de la période de suivi allait de 6 mois à 22 mois (moyenne: 11 mois). Il n'y a pas eu d'incidents de perte du lambeau, de difformité en marche d'escalier, d'obstruction nasale ou de rétraction de l'aile du nez. Au suivi postopératoire de 3 mois, des cicatrices rouge pâle ont été observées dans la zone opératoire de quelques patients (2/15). Cependant, ces cicatrices sont devenues progressivement invisibles à la visite postopératoire de 6 mois. Tous les patients (ou leurs parents) ont été satisfaits du résultat esthétique de cette opération. Conclusion: Ce lambeau trifolié nouvellement conçu peut être une méthode de substitution pour la reconstruction des anomalies congénitales isolées du pourtour de l'aile du nez chez des patients pédiatriques. Les cicatrices secondaires à cette opération peuvent être non évidentes avec une suture chirurgicale fine.

Molecular mechanism analysis of Miao medicine Jinwujiangu decoction in treating osteoarthritis based on a network pharmacology approach.

OBJECTIVE: To investigate molecular mechanisms of Jinwujiangu decoction (, JWJG) in treating osteoarthritis (OA) using network pharmacology analysis. METHODS: Principal active compounds of JWJG were screened out via the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and BATMAN-TCM, and potential targets for OA treatment were identified through Online Mendelian Inheritance in Man (OMIM) and GeneCards databases. The JWJG network diagrams of both principal chemical compound-action targets and OA treatment target-OA disease were constructed applying the Cytoscape 3.7.2 software. The diagram of protein-protein interaction network was plotted for core analysis. Meanwhile, the common targets and relevant signaling pathways involved in both networks were analyzed using the Gene Ontology function analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment. The predicted results were ultimately verified through animal experiments. RESULTS: Effects of JWJG were indicated in acting on key targets interleukin-6, insulin, protein kinase B, glyceraldehyde-3-phosphate dehydrogenase, and mitosis-specific MRE11-RAD50-NBS1 associated protein by regulating signaling pathways of phosphoinositide 3-kinase- protein kinase B, mitogen-activated protein kinases, tumor necrosis factor, and colorectal cancer. Meanwhile, it inhibited the over-activation of signaling pathways and the release of inflammatory factors in OA treatment. Following a signaling pathway analysis utilizing network pharmacology technique, it was demonstrated that JWJG could treat OA through the Wnt/ß-catenin signaling pathway verified by animal experiments. CONCLUSIONS: The present study preliminarily analyzed the pharmacological mechanism of JWJG in treating OA on a network pharmacology approach and provided a great theoretical significance for clinical application.

Psychological Adjustment During Outbreak of COVID-19 Based on Traditional Chinese Medicine Emotion Therapy / 中国实验方剂学杂志

The outbreak of corona virus disease-2019 (COVID-19) not only endangers the public life and health, but also poses threats to the public mental health. Under the atmosphere of tension and fear, the public often produces adverse emotional reactions to psychological stress, and even has extreme behaviors, which seriously affects physical and mental health. Traditional Chinese medicine （TCM） emotional therapy is based on the TCM consciousness theory in adherence to the holistic view of "integration of mind and body". It adopts safe and effective emotional conditioning methods to regulate mood, eliminate bad emotions, maintain mental health, and enhance the body's immunity to achieve the effect of treating or preventing physical and mental diseases. It has differentiated applications in many medical fields, such as post-traumatic stress disorder, generalized anxiety disorder, cancer and hospice, with good clinical efficacy. By analyzing the theoretical origin, research status, application range of TCM sentiment therapy, as well as the current public psychological state of the COVID-19 epidemic, we innovatively proposed TCM sentiment therapies, such as calm emotion method, diversion of attention method, suggestive induction method, obedient mind method, music delight method, TCM refreshing method and auricular pressing method, and comprehensively applied them in the public psychological adjustment during the outbreak, with the aim to provide rational and effective psychological guidance, help the public to correctly understand various bad emotions during the outbreak, enhance their abilities to cope with it and adjust psychological problems, maintain a good mentality, and pass the epidemic period smoothly. At the same time, in order to make better use of the advantages of TCM sentiment therapy and benefit the public, it is advocated to widely apply TCM sentiment therapy in similar public health events and public psychological care in the daily life, for the purpose of preventing before disease outset and preventing progress of disease, and effectively protecting the physical and mental health of the people.

Mechanism of Inflammatory Storm Induced by Novel Coronavirus Pneumonia and Intervention Measures of Traditional Chinese Medicine / 中国实验方剂学杂志

Some patients with corona virus disease-2019 (COVID-19) experienced a severe exacerbation of the disease due to the occurrence of inflammatory storm during the development of the disease. They are complicated with acute respiratory distress, multiple organ dysfunction syndrome and other serious complications, with a poor prognosis and a high mortality. For the inflammation storm, western medicine mostly adopts glucocorticoids, nutritional support, artificial ventilation assistance and other measures at present. The development of artificial liver, blood purification therapy, extracorporeal membrane oxygenation and other technologies have also reduced the mortality of patients to some extent. However, due to the high requirements for equipment, the measures have not yet been widely carried out. From the perspective of traditional Chinese medicine(TCM), the basic pathogenesis of COVID-19 is epidemic toxin invasion, lung and spleen being affected by pathogens, damaging vital Qi, and pathological properties involving dampness, heat, poison, stasis and deficiency. At the stage of inflammation storm, the pathogens are abundant, while the vital Qi is deficient， and the pathogens occlude lung, and disturb the heart and mind, and blood stasis and toxicity are combined with Qi-Yin deficiency. In severe cases, even both Yin and Yang exhaustion occurs. At present, a number of studies have shown that a variety of Chinese herbal medicines have multi-target immunomodulatory effect on viral pneumonia and cytokine storm. TCM participates in whole process of the occurrence and development of inflammation storm, mainly eliminating pathogens in early stage, controlling inflammation and blocking occurrence of inflammation storm， eliminating pathogens and strengthening the body resistance to eliminate the pathological products of the inflammation storm, and promoting the dissipation and absorption of inflammation in the middle stage， and saving lives in late stage by benefiting Qi and relieving depletion, and restoring Yang and rescuing from collapse. On the basis of pathophysiological mechanism of COVID-19 inflammation storm and the theory of TCM syndrome differentiation and treatment, this paper summarized the pharmacological studies on intervention on inflammatory storm with relevant Chinese herbal medicine, Chinese medicine prescriptions and Chinese medicine preparations, and discussed the intervention measures of TCM in different development stages of inflammatory storm, in expectation of providing the guidance for clinical treatment.

Syndrome Differentiation-based Treatment of COVID-19 Based on Theory of Wenyilun / 中国实验方剂学杂志

The outbreak of corona virus disease-2019 (COVID-19) in Wuhan, China, in December 2019, is highly contagious and epidemic, and belongs to the category of "plague" in traditional Chinese medicine, which seriously threatens human life and health. In the face of this sudden public health event, based on the theory of Wenyilun, this paper analyzes the onset characteristics, transmission, and syndrome differentiation-based treatment of COVID-19 in clinical practice. According to the characteristics of this epidemic, the pathogeny of COVID-19 is "epidemic pathogenic factors", and the pathogenesis is that the epidemic suppresses the dampness. Its transmission follows the regularity of "nine transmissions between exterior and interior" recorded in Wenyilun, which can be divided into sequential transmission and reverse transmission. The course of the disease has certain regularity and stages. In the early stage, the pathogen hides in the pleurodiaphragmatic interspace, and the epidemic suppresses the dampness， the treatment shall reach the pleurodiaphragmatic interspace, clear the dampness and turbidness, diffuse the lungs and dredging the lung meridian. In the middle stage, the pathogen can be transmitted to the exterior， the treatment shall be pungent in flavor and cool in property and could relieve the exterior syndrome, so as to remove the pathogen by shivering and sweating. If the dampness toxin enters the interior and be transmitted into the heat, and caused damage to the lung meridian, obstruction of Qi in the lungs, interior excess and heat accumulation, the treatment shall dredge bowel and purge heat, and expel the pathogen out. Any delay in treatment, mistreatment or excessive pathogen can lead to pathogenic invasion into the pericardium, and cause delirium, dysphoria, reversal cold of limbs, barely palpable pulse and other critical manifestations of internal blocking causing external depletion. The treatment shall focus on both attack and supplement, open blocking and relieve depletion. In the recovery period, due to the residual pathogen and the Yin and Qi impairment caused by fever, the treatment shall prevent the recurrence of the residual pathogen, nourish Yin and blood, and restore the vital energy. During the treatment, WU You-ke put forward three principles of "removing the pathogen as the priority, and then avoiding false sweating, and caring for the spleen and stomach". The theory of Wenyilun has a systematic understanding of the pathogeny, pathogenesis, and syndrome differentiation-based treatment of "the epidemic", and provides valuable experience for fight against plagues in later generations, which is also of great significance for the prevention and treatment of COVID-19.

Discussing Differences of Different Traditional Chinese Medicine Schemes of COVID-19 Based on Principle of "Treatment in Accordance with Three Categories of Etiologic Factors" / 中国实验方剂学杂志

Since the outbreak of corona virus disease-2019 (COVID-19), the timely intervention of Chinese medicine has played an important role， it can significantly improve the clinical symptoms of patients, alleviate fever, cough, shorten the course of disease and reduce the conversion rate of mild disease to severe disease， moreover, it has obvious advantages in improving survival rate, prognosis and quality of life in patients. In order to make better use of the therapeutic effect of Chinese medicine and overcome the epidemic situation as soon as possible, various provinces, municipalities and autonomous regions have introduced local Chinese medicine treatment schemes in accordance with the geography, climate and epidemic situation. In this article, we have analyzed the similarities and differences among treatment schemes from the perspective of three factors, and it is believed that the onset season is the same for this disease, and the differences between treatment schemes mainly depend on geographical and climatic factors. The results showed that in seven regions of China, the early symptoms in warm and humid regions such as Central China, South China, East China and Southwest China were characterized by "wet, warm, hot, and toxic", and treatment schemes mainly adopted the recipes with clearing heat and detoxifying, strengthening spleen and removing dampness functions. The early symptoms in cold regions such as Northeast China, North China, and Northwest China were mostly characterized by "cold, wet, wind, and toxic", and the therapeutic recipes emphasized on the effects of expelling wind and cold, removing toxicity and dampness. Among them, the national plan with the Hubei epidemic situation as an important reference has gradually evolved from a single syndrome attribute of each period to both cold and hot syndromes with multiple treatment methods. With the development of the disease, epidemic toxin and toxic heat in the lungs are the main characteristics in the medium term, when the treatment mainly focuses on purging heat and removing toxins. In severe cases and advanced stage, internal block and outward desertion are the main symptoms, when the treatment mainly focuses on block dredging and relieving desertion. In the recovery period, deficiency of lung and spleen Qi, and deficiency of Qi and Yin are the main symptoms, when the treatment is mainly used to invigorate the spleen, tonify the lungs, and nourish Qi and Yin. The physical factors are mostly reflected in the prevention program, so they are rarely mentioned in the treatment program. The Chinese medicine treatment schemes of COVID-19 in different regions show obvious differences. It is of great significance to analyze these differences and sum up the corresponding laws for the differential prevention and treatment of infectious diseases like COVID-19.

Effect of Paiteling on HeLa Cell Proliferation and Metastasis Ability and PI3K/Akt Signal Transduction Pathway / 中国实验方剂学杂志

Objective:To explore the effect of Paiteling on the proliferation，metastasis and invasion of HeLa cells and relevant proteins of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. Method:① HeLa cells were divided into blank group and Paiteling concentration gradient groups (3.906，2.604，1.953，1.563，1.302，1.116，0.977 g·L-1). After drug intervention for 24 h，the cell morphological changes were observed under microscope. The cell viability was measured by thiazole blue (MTT) colorimetry，and the half inhibitory concentration (IC50) of Paiteling on HeLa cells was calculated. ② HeLa cells were divided into blank group，cisplatin group (0.01 g·L-1)，Paiteling high-dose group (2.974 g·L-1)，Paiteling medium-dose group (1.487 g·L-1) and Paiteling low-dose group (0.991 g·L-1). Cell proliferation and toxicity test (CCK-8) method was used to detect the effect of Paiteling on the proliferation ability of HeLa cells，scratch test was used to detect cell migration，and invasion test (Transwell) was used to detect changes in cell invasion ability. ③ Inhibitor LY294002 group (0.006 g·L-1) was added. Western blot (WB) was used to detect the expressions of Paiteling on PI3K，Akt，recombinant human B-cell lymphoma factor-xl (Bcl-xl)，and B-cell lymphoma/leukemia associated D protein (Bad). Result:① Compared with the blank group，microscopic observation showed that the number of cells in the treatment group was significantly reduced, and the cell morphology was incomplete. MTT experiments showed that Paiteling has a significantly inhibitory effect on HeLa cell proliferation (P<0.01). The IC50 of Paiteling on HeLa cells was calculated as 2.974 g·L-1. ② The CCK-8 experiment showed that compared with the blank group，all the drug-treated groups had an inhibitory effect on HeLa cell proliferation at 24，36，48 h (P<0.01), compared with the cisplatin group，middle and low-dose Paiteling groups showed a reduced inhibitory effect on HeLa cell proliferation at each time point (P<0.01). The scratch test showed that，compared with the blank group，each drug-added group could inhibit the migration ability of HeLa cells (P<0.01)，and the cell migration rate of the high-dose Paiteling group was lower than that of the cisplatin group (P<0.05). Transwell experiments showed that compared with the blank group，the number of membranes permeated by HeLa cells in each drug-treated group was decreased (P<0.01)，and the number of membranes permeated in the middle and low-dose Paiteling groups was increased compared with the cisplatin group (P<0.01). ③ Western blot showed that compared with the blank group，the expression levels of PI3K，Bcl-xl，and Akt in the high，medium，and low-dose Paiteling groups and the LY294002 group decreased (P<0.05，P<0.01)，while the expression of Bad increased (P<0.01). Compared with the high-dose Paiteling group，the PI3K，Akt，and Bcl-xl protein expressions were increased in the low-dose Paiteling group (P<0.01)，whereas Bad expression was decreased (P<0.01). Conclusion:Paiteling can inhibit HeLa cell proliferation，metastasis and invasion ability in a dose-dependent and time-dependent manner，which may be related to its effect on the expressions of PI3K/Akt signaling pathway-related proteins.

Molecular Mechanism of Tangshenping Capsule in Intervening Apoptosis of Podocytes Induced by High Glucose+LPS Through PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective: To observe effect of Tangshenping capsule on lipopolysaccharide (LPS)-stimulated podocytes apoptosis under high glucose conditions of podocyte in rats with diabetic nephropathy (DN), and discuss possible mechanism. Method: With cultured rat podocytes as object of study,the podocytes model was established with high glucose and LPS,and divided into 7 groups:control group,high glucose group,high glucose plus LPS group,irbesartan group,and low,moderate and high-dose Tangshenping capsule groups.Protein and mRNA expressions of CD2-associated protein (CD2AP), nephrin, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and B-cell lymphoma-2 associated death protein (Bad protein) of podocyte in each group were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR). Result: Compared with control group,the protein and mRNA expressions of podocyte CD2AP, nephrin, PI3K, Akt decreased definitely (PPPPPPPPPPPPConclusion: Tangshenping capsule can maintain a stable protein expression of Slit diaphragm (SD) in podocyte, inhibit podocyte apoptosis by activating PI3K/Akt signaling pathway. This is one of mechanisms for preventing and treating diabetic nephropathy.

Renal Protective Effect of Tangshiping Capsule on Diabetic Nephropathy Kkay Mice and Its Effect on Wnt/β-catenin Signal Transduction Pathway / 中国实验方剂学杂志

Objective:To investigate the renal protective effect of Tangshenping capsule (Tangshenping) on diabetic nephropathy (DN) KKAy mice and its effect on Wnt/β-catenin signaling pathway. Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks. The DN animal model was successfully determined with blood glucose (>16.7 mmol·L-1) and 24 hour urine protein (>0.4 mg). The model mice were randomly divided into a model group, an irbesartan group, and low, medium and high-dose Tangshenping group, with 10 female C57BL/6J mice as a control group. The treatment groups were given the corresponding drugs by gavage. The normal group and the model group were given an equal volume of deionized water by gavage. The intragastric dose was 0.01 mL·g-1 body weight coefficient once a day. The general conditions of the mice were observed, the body mass was weighed every 4 weeks, and 24 h urine protein was quantified. At the 26th week, the blood was collected from eyeballs, and the mice were put to death. The quality of the kidneys, serum blood urea nitrogen (BUN), serum creatinine (SCr), triglyceride (TG), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) content were measured. In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4, glycogen synthase kinase 3β(GSK3β) and β-catenin in kidney tissues. Result:Compared with model group, body mass, kidney mass/body mass, and 24 h urine protein were significantly lower in high-dose Tangshenping group (PPPβ and β-catenin were decreased (PConclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway, reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice, delay the progression of renal interstitial fibrosis, and then exert renal protection.

Investigation of the Antioxidant Capacity, Insecticidal Ability and Oxidation Stability of Chenopodium formosanum Seed Extract.

To maximize the extraction of antioxidants from Chenopodium formosanum seeds, the process factors, such as the ethanol concentration (0⁻100%), extraction time (30⁻180 min) and temperature (30⁻70 °C), for the extraction of the bioactive contents as well as the antioxidant capacity are evaluated using response surface methodology (RSM). The experimental results fit well with quadratic models. The extract was identified by GC/MS, and it was found that some active compounds had antioxidant, repellency and insecticidal activities. Various concentrations of the extract were prepared for the evaluation of the insecticidal activity against Tribolium castaneum, and the toxicity test results indicated that the extract was toxic to Tribolium castaneum, with an LC50 value of 354.61 ppm. The oxidative stability of the olive oil determined according to the radical scavenging activity and p-anisidine test demonstrates that the extract obtained from the Chenopodium formosanum seeds can retard lipid oxidation.

Immunogenicity moderation effect of interleukin-24 on myelogenous leukemia cells.

Previous studies have shown that interleukin-24 (IL-24) has tumor-suppressing activity by multiple pathways. However, the immunogenicity moderation effect of IL-24 on malignant cells has not been explored extensively. In this study, we investigated the role of IL-24 in immunogenicity modulation of the myelogenous leukemia cells. Data show that myelogenous leukemia cells express low levels of immunogenicity molecules. Treatment with IL-24 could enhance leukemia cell immunogenicity, predominantly regulate leukemia cells to produce immune-associated cytokines, and improve the cytotoxic sensitivity of these cells to immune effector cells. IL-24 expression could retard transplanted leukemia cell tumor growth in vivo in athymic nude mice. Moreover, IL-24 had marked effects on downregulating the expression of angiogenesis-related proteins vascular endothelial growth factor, cluster of differentiation (CD) 31, CD34, collagen IV and metastasis-related factors CD147, membrane type-1 matrix metalloproteinase (MMP), and MMP-2 and MMP-9 in transplanted tumors. These findings indicated novel functions of this antitumor gene and characterized IL-24 as a promising agent for further clinical trial for hematologic malignancy immunotherapy.

Field-evolved resistance to insecticides in the invasive western flower thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) in China.

BACKGROUND: To understand the current status of insecticide resistance of the invasive western flower thrips, Frankliniella occidentalis, in China, the responses of six field populations to six commonly used insecticides, i.e. spinosad, spinetoram, cyantraniliprole, imidacloprid, acetamiprid and pyriproxyfen, were evaluated in comparison with a susceptible laboratory strain. RESULTS: Field populations tended to be less susceptible than the laboratory strain. The population from Shouguang, Shandong Province, showed the lowest levels of susceptibility. A 15.64-fold and 17.29-fold resistance to spinosad and spinetoram was detected in the Shouguang population. A 11.74-fold and 13.64-fold resistance to cyantraniliprole was detected in populations from Daxing in the Beijing area and Shouguang. All populations showed a low level of resistance to imidacloprid, acetamiprid and pyriproxyfen, except for the Shouguang population, which was 127.58-fold more resistant to pyriproxyfen. CONCLUSION: Variations in resistance to the tested insecticides were observed among the sampled population. Spinosad and spinetoram were the most efficient insecticides and are recommended for use in an integrated management programme. Resistance management strategies should be implemented to reduce the potential for resistance evolving. © 2015 Society of Chemical Industry.

Antitumor effect and underlying mechanism of RGD-modified adenovirus mediated IL-24 expression on myeloid leukemia cells.

Interleukin-24 (IL-24), a member of the IL-10 cytokine gene family, causes growth suppression and apoptosis in various solid tumor cells. However, the effects of IL-24 on hematopoietic malignant cells have not been extensively explored. In this report, we constructed an RGD-engineered recombinant adenoviral vector, Ad.RGD-IL-24, and assessed its effects on human myeloid leukemia cells. Ad vector mediates gene transfer into leukemia cells with high efficiency. Ectopic over-expression of IL-24 has significant growth inhibition and differentiation inducement effects on these cells. Treatment with Ad.RGD-IL-24 could potentially induce leukemia cells' cell-cycle arrest. In addition, IL-24 expression could significantly induce apoptosis of the THP-1 cells. Ad.RGD-IL-24 had a potent effect on the up-regulation of the expression of GRP78/Bip, GADD34 and Bax, down-regulation of the expression of Bcl-2 and Mcl-1, and induced the activation of Caspase-3, which may be responsible for its apoptosis-inducing effect on THP-1 cells. Furthermore, IL-24 expression could retard transplanted leukemia cell tumor growth in vivo in athymic nude mice. These findings showed the marked antitumor activity of IL-24 and provided potential perspectives in designing therapeutic or vaccine strategies in immuno-gene therapy of myeloid leukemia.

Preparation and characterization of monoclonal antibody against human B7-H4 molecule.

B7-H4, a member of B7 family, is widely expressed in tumor tissues and plays an important role in the negative regulation of T cell immunity. In this study, we report on the establishment and characterization of a functional anti-human B7-H4 monoclonal antibody (MAb) 5G3 through hybridoma method. Flow cytometry analysis showed that MAb 5G3 specifically bound to B7-H4 molecule. Functional experiments indicated that MAb 5G3 could block the inhibitory role of B7-H4 molecule on A549 cells in and reduce the apoptosis of Jurkat cells, suggesting that MAb 5G3 is an antagonistic antibody and a useful tool for further studies of B7-H4 functions in cancers.

Expression heterogeneity research of ITGB3 and BCL-2 in lung adenocarcinoma tissue and adenocarcinoma cell line.

OBJECTIVE: To analyze expression heterogeneity of Integrin beta 3 (ITGB3) and B-cell lymphoma 2 (BCL-2) in lung adenocarcinoma tissue and adenocarcinoma cell line and further provide theoretical direction for molecular biological research of lung adenocarcinoma. METHODS: Tissue microarray was used to observe relation among expression, heterogeneitpy and clinical characteristics of ITGB3 and BCL-2 in lung cancer. RESULTS: ITGB3 and BCL-2 increased significantly in A549 cells in CAFs group withß-actin as control; the expression level of BCL-2 also increased in ITGB3 transfected cells with GFP plasmid transfected A549 cells as control; immunohistochemistry staining showed that positive rates of ITGB3, ITGB1 and BCL-2 in normal lung tissues were 0, the positive rates in lung adenocarcinoma were 7.04%, 84.51% and 4.23%, respectively; in the results of immunohistochemistry staining, the expression of Girdin protein in lung adenocarcinoma was homogeneous, however protein expression of ITGB3, ITGB1 and BCL-2 showed different patterns in the same location with significant heterogeneity; majority of ITGB3, ITGB1 or BCL-2 positive tissue showed heterogeneity that expression in trailing edge was higher than that of trailing edge in lung adenocarcinoma tissue, the patients with BCL-2 heterogeneity showed higher lymph node metastasis ratio and lower clinical stage (P<0.05); and the expression of ITGB3 and the clinical characteristics of patients were not significant related (P>0.05). CONCLUSIONS: Expression of ITGB3 and BCL-2 in lung adenocarcinoma and adenocarcinoma cell line showed heterogeneity that expression in trailing edge was higher than that of trailing edge, which may play an important role in promoting tumor lymph node metastasis and vascular invasion, and provides a new research direction for exploration of lung adenocarcinoma metastasis mechanism.

Expression heterogeneity research of ITGB3 and BCL-2 in lung adenocarcinoma tissue and adenocarcinoma cell line

OBJECTIVE@#To analyze expression heterogeneity of Integrin beta 3 (ITGB3) and B-cell lymphoma 2 (BCL-2) in lung adenocarcinoma tissue and adenocarcinoma cell line and further provide theoretical direction for molecular biological research of lung adenocarcinoma.@*METHODS@#Tissue microarray was used to observe relation among expression, heterogeneitpy and clinical characteristics of ITGB3 and BCL-2 in lung cancer.@*RESULTS@#ITGB3 and BCL-2 increased significantly in A549 cells in CAFs group withβ-actin as control; the expression level of BCL-2 also increased in ITGB3 transfected cells with GFP plasmid transfected A549 cells as control; immunohistochemistry staining showed that positive rates of ITGB3, ITGB1 and BCL-2 in normal lung tissues were 0, the positive rates in lung adenocarcinoma were 7.04%, 84.51% and 4.23%, respectively; in the results of immunohistochemistry staining, the expression of Girdin protein in lung adenocarcinoma was homogeneous, however protein expression of ITGB3, ITGB1 and BCL-2 showed different patterns in the same location with significant heterogeneity; majority of ITGB3, ITGB1 or BCL-2 positive tissue showed heterogeneity that expression in trailing edge was higher than that of trailing edge in lung adenocarcinoma tissue, the patients with BCL-2 heterogeneity showed higher lymph node metastasis ratio and lower clinical stage (P0.05).@*CONCLUSIONS@#Expression of ITGB3 and BCL-2 in lung adenocarcinoma and adenocarcinoma cell line showed heterogeneity that expression in trailing edge was higher than that of trailing edge, which may play an important role in promoting tumor lymph node metastasis and vascular invasion, and provides a new research direction for exploration of lung adenocarcinoma metastasis mechanism.

Preferential expansion of umbilical cord blood-derived CD34-positive cells on human leukemia inhibitory factor transgenic feeder cells cultured on regenerated silk fibroin film.

In vitro expansion of transplantable hematopoietic stem cells (HSCs) is a very promising approach for different clinical applications. We have recently developed a new culture system that facilitates in vitro expansion of transplantable cord blood HSCs. In our study, we constructed a recombinant adenovirus Ad-GFP/human leukemia inhibitory factor (hLIF) expressing hLIF. The hLIF gene was delivered into human embryo lung fibroblast cell line WI-38 via infection with Ad-GFP/hLIF. Then, the transgenic cells were cultured on regenerated silk fibroin (SF) films as feeder layer cells for expansion of cord blood CD34(+) cells. Our results showed that the hLIF transgenic WI-38 cells cultured on SF could express hematopoiesis-related cytokines at higher levels compared with control groups. The hLIF-expressing feeder layer cells cultured on SF in combination with cytokines more efficiently expanded CD34(+) cells and CD34(+) CD38(-) cells. The percentages of adhesion molecules on the expanded CD34(+) cells in transgenic feeder layer cells cultured on SF were higher than those of control groups. Interestingly, the migration rate assessed by transwell assay was also significantly higher than those of control groups, which suggests that transgenic feeder layer cells cultured on SF has powerful ability to maintain the homing capacity of expanded CD34(+) cells.

Correlation between pesticide resistance and enzyme activity in the diamondback moth, Plutella xylostella.

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is one of the most important pests that has developed high pesticide resistance. The resistances of five Chinese populations of this moth, four resistant strains (from Beijing, Henan, Fujian, and Guangdong) and one susceptible strain, to five pesticides were determined, and the activities of carboxylesterase, glutathione S-transferase, and acetylcholine esterase were tested in all five populations. The correlations between pesticide resistance and enzyme activity were analyzed. The results showed that the resistance status to the five pesticides was different among the five populations. The resistance ratios of the Beijing and Henan populations to spinosad were 5.84 and 8.22, respectively, and those to beta-cypermethrin were 4.91 and 4.98, respectively. These ratios were higher than those for the Fujian and Guangdong populations. The Fujian population was more sensitive to abamectin and chlorpyrifos than the susceptible population (the resistance ratios were 0.14 and 0.91, respectively); in fact, the median lethal concentration for P. xylostella was significantly higher for chlorpyrifos than that for any of the other four pesticides. The carboxylesterase activity in P. xylostella showed positive correlations with the resistance to spinosad, beta-cypermethrin, chlorpyrifos, and abamectin, but no correlation was observed between the carboxylesterase activity and resistance to emamectin benzoate, between glutathione S-transferase activity and resistance to any of the five pesticides tested, or between acetylcholine esterase activity and any of the pesticides except for emamectin benzoate.

[Epitopic multiple antigen peptide from α-fetoprotein elicit antitumor immune response in vitro and ex vivo].

OBJECTIVE: To explore the antitumor effects of multiple antigen peptide (MAP) vaccine from α-fetoprotein (AFP) through AFP-specific cytotoxic T lymphocyte (CTL) against hepatoma in vitro and ex vivo. METHODS: Dendritic cells (DC) were generated from human peripheral blood mononuclear cells (PBMC) and HLA-A2.1-transgenic murine bone marrow. The AFP-specific CTL were induced by MAP-loaded DC and the corresponding linear peptides from human AFP. The lysis rate of effectors to hepatoma cells were tested by 4 h (51)Cr release assay. And enzyme-linked immunosorbent spot (ELISPOT) was used to test the interferon (IFN)-γ release of effector cells. RESULTS: The specific lysis rate of effectors induced by AFP epitopic MAP vaccines to Hep3B cells (AFP(+), HLA-A2.1(+)) at the highest effector/target (E/T) ratio was significantly higher than linear peptide vaccine (73.5% ± 7.9% vs 45.6% ± 6.9%, P < 0.01). The effectors induced by AFP epitopic MAP vaccine and linear peptide vaccine could not lysis the AFP-negative PLC/PRF/5 liver cancer cells versus the negative control group at the highest E/T (9.3% ± 3.9%, 8.1% ± 2.8% vs 8.3% ± 2.6%, both P > 0.05). But the effectors induced by AFP epitopic MAP vaccine and linear peptide vaccine could lyse PLC/PRF/5 liver cancer cells transfected with cDNA of AFP versus the negative control group (74.8% ± 10.5%, 51.4% ± 12.6% vs 4.2% ± 1.3%, both P < 0.01). And the specific lysis rate of effectors induced by AFP epitopic MAP vaccines was significantly higher than the corresponding linear peptide vaccine (P < 0.01). Compared with the negative control group, the effectors could not lyse HepG2 liver cancer cells, a HLA-A2.1 negative cell line (both P > 0.05). But the effectors could lyse HepG2 cells transfected with cDNA of HLA-A2.1 (71.8% ± 8.6%, 46.5% ± 6.5% vs 4.1% ± 1.1%, both P < 0.01). And the specific lysis rate of effectors induced by MAP vaccine was significantly higher than the corresponding linear peptide vaccine (P < 0.01). ELISPOT test showed that the capability of enhancing IFN-γ release of human AFP MAPs was stronger than that of the AFP linear peptides. The spots count of MAP vaccine group ((158 ± 23) spots/10(5) cells) or linear peptide vaccine group ((78 ± 12) spots/10(5) cells) were significantly higher than the negative control group ((3 ± 1) spots/10(5) cells) (all P < 0.01). The spots count of the positive control group ((166 ± 32) spots/10(5) cells) showed no significant difference with the AFP MAP vaccine group (P > 0.05). And the spots count of MAP vaccine group were significantly higher than the corresponding linear peptide vaccine group ((78 ± 12) spots/10(5) cells, P < 0.01). CONCLUSIONS: AFP multiple antigen peptides elicit not only more powerful specific anti-tumor immune responses but also stronger non-specific anti-tumor immune activities than their corresponding linear peptides. These findings will provide theoretical rationales for their clinical applications.

The complete mitochondrial genome of the oriental fruit moth Grapholita molesta (Busck) (Lepidoptera: Tortricidae).

The oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae) currently is one of the economically most destructive pest species of stone and pome fruits worldwide. Here we sequenced the complete mitochondrial genome of this pest. This genome is 15,776 bp long, with an A + T content of 81.24%, containing 37 typical animal mitochondrial genes and an A + T-rich region. All gene are arranged as hypothesized ancestral gene order of insects except for trnM, which was shuffled from 3' downstream of trnQ to 5' upstream of trnI. cox1 gene uses unusual CGA start codon, as that in all other sequenced lepidopteran mitochondrial genome. The secondary structures for the two rRNA genes were predicted. All helices typically present in insect mitochondrial rRNA genes are generated. A microsatellite sequence was inserted into the region of H2347 in rrnL in G. molesta and two other sequenced tortricid mitochondrial genomes, indicating that the insertion event in this helix might occurred anciently in family Tortricidae. All of the 22 typical animal tRNA genes have a typical cloverleaf structure except for trnS2, in which the D-stem pairings in the DHU arm are absent. An intergenic sequence is present between trnQ and nad2 as well as in other sequenced lepidopteran mitochondrial genomes, which was presumed to be a remnant of trnM gene and its boundary sequences after the duplication of trnM to the upstream of trnI in Lepidoptera. The A + T-rich region is 836 bp, containing six repeat sequences of "TTATTATTATTATTAAATA(G)TTT."