OBJECTIVETo construct an injectable tissue-engineered bone graft with fibrin glue (FG), autologous platelet-rich plasma (PRP) and bone marrow stromal cells (BMSCs) cultured in vitro and study its biological characteristics and microscopic structures.

METHODSBMSCs isolated from rabbit iliac bone marrow were culture-expanded in vitro. The injectable tissue-engineered bone constructed from autologous PRP, FG, and BMSCs was cultured in vitro, and its biological characteristics were observed including the time of gel formation, histological features, seed cell survival and microscopic structures.

RESULTSThe constructed injectable tissue-engineered bone began gel formation within 20 to 30 s, and after a week-long culture, the gelatine began to degrade, and numerous well viable fusiform cells could be seen to adhere to the bottom of the Petri dish. Scanning electron microscopy identified globular and olivary cells embedded in the fibrin glue, and numerous small particles could be seen around of the cells.

CONCLUSIONConstruction of an injectable tissue-engineered bone graft with FG, BMSCs and PRP does not require sophisticated techniques and ensures good biological property of the bone graft that can be easily shaped and allow good growth of the seed cells, suggesting great potential of this technique for clinical use.

Bone Marrow Cells ; cytology ; Bone Substitutes ; chemistry ; Cells, Cultured ; Coculture Techniques ; Fibrin Tissue Adhesive ; chemistry ; Platelet-Rich Plasma ; chemistry ; Stromal Cells ; cytology ; Tissue Engineering ; methods

OBJECTIVESTo investigate the engraftment, survival and graft-versus host disease (GVHD) after transplantation of unrelated cord blood for the treatment of childhood and adult hematological malignancies.

METHODSSeventeen patients (13 children and 4 adults) with hematological malignancies were enrolled in this study. Twelve patients were transplanted with one unit and 5 with 2 units of cord blood. There were HLA-matched in 6 and HLA-mismatched at 1 approximately 2 loci in 11 patients. Ten patients were transplanted at stable status, 7 at advanced stage of leukemia. Conditioning regimens were BU/CY for 13 and CY/TBI for 3 patients. Most patients received additional ATG at a dose of 15 approximately 20 mg x kg(-1) x d(-1) for 3 days. CsA, mycophenolate mofetil (MMF) and methylprednisolone were used for GVHD prophylaxis.

RESULTSFourteen patients survived more than 40 days after transplantation were evaluated for engraftment. At day 60 after UCBT, 86% and 71% of the patients showed neutrophil and platelet engraftment, respectively. The time for an absolute neutrophil count > or = 0.5 x 10(9)/L was (21.0 +/- 1.3) days and platelet > or = 20 x 10(9)/L was (39.0 +/- 10.3) days. Four patients developed grade II acute GVHD and 2 chronic GVHD. Of the 17 patients, 11 were still alive and 8 of them were in event-free status. For the 10 patients transplanted at stable status, 2 year overall survival is 90%, and event-free survival (EFS) 70%. However, for the 7 patients transplanted at advanced stage of leukemia, only 2 survived without relapse. Of the 4 adult patients, 2 had sustained engraftment and survived for 18 and 14 months, respectively.

CONCLUSIONSHLA-matched or 1 approximately 2 loci-mismatched UCBT is a feasible procedure to cure a significant proportion of children or adults with leukemia, especially if performed in a favourable phase of disease. Two units of CBT can be used for adult patients if the cell number of one unit is not enough.

Adolescent ; Adult ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; etiology ; Hematologic Neoplasms ; immunology ; mortality ; therapy ; Histocompatibility Testing ; Humans ; Male ; Survival Analysis ; Survival Rate ; Transplantation Conditioning

As a treasure of traditional Chinese medicine culture,Dao-di herbs are famous for their high quality and good effect.However,traditional characteristics of Dao-di herbs and their producing areas are mostly confined to qualitative description,lacking objective evaluation indicators. Numerical taxonomy,which uses mathematical methods to improve the research of taxonomy from qualitative description to quantitative comprehensive analysis,is objective and accurate,so is widely used in the field of biology and medicine.This paper reviews the concept and main research fields of numerical taxonomy,and summarizes its application status and development prospects in the study of Dao-di herbs and characteristics of production areas,in order to lay the foundation for the objectification and standardization of Dao-di herbs.
Drugs, Chinese Herbal ; classification ; standards ; Medicine, Chinese Traditional ; Plants, Medicinal ; classification

Geranylgeranyl pyrophosphate synthase enzyme is one of the key enzymes in the synthesis pathway of diterpenoid. Nine Lamiaceae genus GGPS synthase in Genebank was analyzed in this article. GGPS synthase the nucleic acid sequences and amino acid sequences, physicochemical properties, the signal peptide, leader peptides, transmembrane topological structure, hydrophobic, hydrophilic, subcellular localization, secondary structure, function domain, tertiary structure and evolutional relationship were predicted by using bioinformatics methods.Phylogenetic tree was constructed for the geranylgeranyl pyrophosphate synthase enzyme protein family. The results showed that GGPS amino acid sequence of the physical and chemical properties were basically identical, mainly hydrophilic protein, there existed chloroplast transit peptide, and no signal peptide and membrane structure domain, which mainly located in the chloroplast, the minor part located in mitochondria. The main secondary structures of the proteins are alpha helix and random coil. All these proteins have catalytic residues, aspartate-rich region, active site lid residues, substrate-Mg2+ binding site. The results provide theoretical reference for study on both the enzymatic characteristics of GGPS and the biosynthesis pathway of diterpenoid.

The Lycium genus consists of consist of 7 species and 3 variety which are main distributed in Northwest region in China, the cultivated Goji berry appeared about 1 000 years ago. The phylogeny of the wild Goji berry and the domestication of cultivated Goji berry are important scientific and practical value due to the medicinal and economic value. In this paper, a new advances achieved in studies on the phylogeny of wild Goji berry is summarized. The origin of cultivated Goji berry includes when, where, how and ancestral specie were reviewed. The current situation of cultivation and existing problems were summarized. We considered that through mutations and artificial selections, wild ancestors were domesticated to current cultivar groups. To find direct evidence for the origin of cultivated Goji berry, we must rely on genetic analysis in addition to morphological characters. To make full use of the Goji berry germplasm resources, the investigation and mining on wild Goji berry resource should be strengthened in the future.

The basic features of glandular and non-glandular trichomes on leaves of Artemisia argyi( germplasms from Qichun,Ningbo,Tangyin,and Anguo,respectively) and related species A. stolonifera were observed by scanning electron microscopy( SEM)and compared. There were significant differences in trichome characteristics of leaves at all parts of A. argyi and A. stolonifera,which were closely related to the difference in chemical components. The length of non-glandular trichomes and size of glandular trichomes on middle leaves were the stablest. A. argyi and A. stolonifera can be distinguished by the density of glandular trichome. Additionally,the four germplasms of A. argyi can be discriminated via the density and curvature of non-glandular trichome. The density of non-glandular trichomes was the highest in A. stolonifera. For A. argyi,the germplasm from Qichun had the highest density of non-glandular trichomes on the abaxial surfaces of upper leaves and that from Ningbo had the largest non-glandular trichome curvature. With regard to the germplasm from Anguo,the T-shaped non-glandular trichomes of long stalks on the adaxial surfaces of the middle leaves were lodging-susceptible,and those with slender heads were wave-like. Statistics results of A. argyi and A. stolonifera are as follows: largest glandular trichomes on the adaxial and abaxial surfaces and highest glandular trichome density on the abaxial surfaces of the lower leaves in A. argyi germplasm from Ningbo,highest density of non-glandular trichomes on the abaxial surfaces of upper leaves in A. stolonifera,and highest density of glandular trichomes and non-glandular trichomes on the adaxial surfaces of the upper leaves in A. argyi germplasm from Qichun. According to the observation result under fluorescence microscope( FM),flavonoids were closely related to the size and density of non-glandular trichomes and size of glandular trichomes. The fluorescence intensity was the strongest and fluorescence area was the largest for flavonoids in A. argyi germplasms from Qichun and Tangyin,while the fluorescence for flavonoids was the weakest in A. stolonifera. It was the first time to observe and analyze the trichome ultrastructure of A. argyi leaves at different positions by SEM and FM. This study clarifies the differences between A. stolonifera and four famous A. argyi germplasms,which provides new evidence for the microscopic identification of A. argyi and its related species and serves as a reference for the study of the relationship of A. argyi structure with its components and functions.
Artemisia ; Flavonoids ; Microscopy, Electron, Scanning ; Plant Leaves ; Trichomes

OBJECTIVE: To investigate the risk factors for brain injury in preterm infants by a multicenter epidemiological investigation of brain injury in hospitalized preterm infants in Anhui, China. METHODS: Preterm infants who were hospitalized in the department of neonatology in 9 hospitals of Anhui Neonatal Collaboration Network between January 2016 and January 2017 were enrolled as subjects. The data of maternal pregnancy and clinical data of preterm infants were collected, and the logistic regression model was used to analyze the risk factors for brain injury in preterm infants. RESULTS: A total of 3 378 preterm infants were enrolled. Of the 3 378 preterm infants, 798 (23.56%) had periventricular-intraventricular hemorrhage (PVH-IVH), and 88 (2.60%) had periventricular leukomalacia (PVL). Intrauterine distress, anemia, hypoglycemia and necrotizing enterocolitis (NEC) were risk factors for PVH-IVH (OR=1.310, 1.591, 1.835, and 3.310 respectively; P<0.05), while a higher gestational age was a protective factor against PVH-IVH (OR=0.671, P<0.05). PVH-IVH, NEC and mechanical ventilation were risk factors for PVL (OR=4.017, 3.018, and 2.166 respectively; P<0.05), and female sex and use of pulmonary surfactant were protective factors against PVL (OR=0.514 and 0.418 respectively; P<0.05). CONCLUSIONS Asphyxia/anoxia, infection/inflammation, mechanical ventilation, anemia and hypoglycemia may increase the risk of brain injury in preterm infants.
Brain Injuries ; Cerebral Hemorrhage ; China ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Premature ; Leukomalacia, Periventricular

Volatile oil is the main effective component and an important quality indicator of Artemisia argyi leaves. In this study, 100 germplasm resources of A. argyi were collected from all the related habitats in China. The total volatile oils in A. argyi leaves were extracted by steam distillation and the content was determined by GC-MS. The result demonstrated that the content of total volatile oils was in the range of 0.53%-2.55%, with the average of 1.43%. A total of 39 chemical constituents were identified from the volatile oils, including 13 shared by the 100 germplasm resources. Clustering analysis of the 39 constituents showed that the 100 A. argyi samples were categorized into groups Ⅰ(9), Ⅱ(2), Ⅲ(66) and Ⅳ(23), and group Ⅲ had the most volatile medicinal components, with the highest content. Five principal components(PCs) were extracted from 13 shared constituents, which explained 73.454% of the total variance. PC1, PC2, and PC3 mainly reflected the pharmacological activity of volatile oils and the rest two the aroma information. The volatile oils identified in this study lay a foundation for variety breeding of and rational utilization of volatile oils in A. argyi leaves.
Artemisia ; Distillation ; Oils, Volatile ; Plant Breeding ; Plant Leaves

italic>Anoectochilus roxburghii liquid (spray, a hospital preparation of Wu Mengchao Hepatobiliary Hospital of Fujian Medical University) has shown a good clinical treatment effect during the COVID-19 pandemic, but its material basis and mechanism of action are still unclear. In this study, network pharmacology and molecular docking methods were used to predict the molecular mechanism of A. roxburghii liquid against COVID-19, and pharmacodynamic experiments in vitro were conducted to study the interaction between the current targets with clear preventive and therapeutic effects and the key components of A. roxburghii liquid. UPLC-MS and database were used to compare and analyze the active ingredients in the liquid, and 17 potential active ingredients with good drug-like properties were screened by in vivo pharmacokinetics process in SwissADME database. SwissTargetPrediction and GeneCards were searched to find 93 common targets. Cytoscape 3.8.2 software was used to construct the "component-target" network map, and the Metascape platform was used for gene function annotation and pathway enrichment analysis. It was found that the extract could regulate the positive response to external stimuli, inflammatory response, cytokine production and other biological processes by binding the active ingredients such as isorhamnetin, kaempferol, luteolin, quercetin and apigenin to the common targets (NOS3, MPO, MMP3, etc.), and play an anti-COVID-19 role. In the angiotensin-converting enzyme 2 (ACE2) activity inhibition assay, it was found that the stock solution of A. roxburghii liquid (for spray), and the supernatant after removing polysaccharides (mainly containing flavonoids) could to some extent inhibit the activity of ACE2. Crucially, in the experiment of 2019-nCOV-S pseudovirus infecting HEK-293T-ACE2 cells, we found that A. roxburghii liquid may exert anti-COVID-19 effects by blocking the binding of SARS-CoV-S protein to ACE2.

Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3-23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.