Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

OBJECTIVE: In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM. METHODS: We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework. RESULTS: We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs. CONCLUSION TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.

Objective To explore the value of low-dose multi-slice spiral computed tomography (MSCT) multi-stage scan and three-dimensional reconstruction in diagnosis of patients with gastric carcinoma.Methods A total of 40 patients with gastric carcinoma were selected.All the patients were conducted with low-dose MSCT scan,dynamic contrast scanning and three-dimensional reconstruction scan,and then were processed by multi-planar reconstruction,volume reconstruction and CT virtual endoscopy.Primary lesions in gastric cancer,peripheral infiltration and distant metastasis were observed.Results All the patients were satisfied with the low dose MSCT images,and all of them found primary gastric lesions and had satisfactory diagnosis rates of gastric peripheral infiltration,lymph node metastasis and distant metastasis.Conclusion Low-dose MSCT multi-stage scan and three-dimensional reconstruction can reduce the radiation dose and reveal the primary gastric lesions,gastric peripheral infiltration and distant metastasis.

Objective To explore the value of low-dose multi-slice spiral computed tomography (MSCT) multi-stage scan and three-dimensional reconstruction in diagnosis of patients with gastric carcinoma.Methods A total of 40 patients with gastric carcinoma were selected.All the patients were conducted with low-dose MSCT scan,dynamic contrast scanning and three-dimensional reconstruction scan,and then were processed by multi-planar reconstruction,volume reconstruction and CT virtual endoscopy.Primary lesions in gastric cancer,peripheral infiltration and distant metastasis were observed.Results All the patients were satisfied with the low dose MSCT images,and all of them found primary gastric lesions and had satisfactory diagnosis rates of gastric peripheral infiltration,lymph node metastasis and distant metastasis.Conclusion Low-dose MSCT multi-stage scan and three-dimensional reconstruction can reduce the radiation dose and reveal the primary gastric lesions,gastric peripheral infiltration and distant metastasis.

Background and purpose:Thyroid carcinoma is a common endocrine tumor with an incidence that has increased over recent decades. The aim of the present study was to investigate the effectiveness of carbon nano-par-ticles-labeled lymph nodes in neck dissection for papillary thyroid cancer (PTC), focusing on the protectiveness for the recurrent laryngeal nerve (RLN) and parathyroid glands.Methods:Forty-eight patients with PTC treated from Apr. to Aug. 2015 were randomly divided into two groups. Group A patients (24 patients) were treated with lobectomy/total thyroidectomy plus unilateral/bilateral central lymph node dissection by conventional meticulous capsular dissection technique; Group B patients (24 patients) were treated with the same surgical procedures as group A, 5 min after the injection of carbon nano-particles. The operative time, intra-operative blood loss, incidence of RLN injury, incidence of transient hypocalcemia, the number of total lymph nodes and the ratio of metastatic nodes were collected and analyzed. Results:For unilateral lobectomy, the number of lymph nodes in group B was signiifcantly greater than that in group A (P<0.05). For total thyroidectomy, the operative time, and the incidence of transient hypocalcemia in group B were both lower than those in group A (P<0.05), and the number of total lymph nodes was signiifcantly higher than that in group A (P<0.05). In group B, the ratio of metastatic nodes were 26.7% (unilateral) and 33.3% (bilateral) in stained lymphnodes, and 11.8% and 25.9% in non-stained lymph nodes.Conclusion:The carbon nano-particles-labeled lymph nodes in neck dissection could facilitate to protect parathyroids and increase the number of lymph nodes, especially in total thyroidectomy plus bilateral central lymph node dissection.

OBJECTIVETo investigate the effect of Ganoderma lucidum polysaccharides (GLPs) on hemodynamic and antioxidation in the T2DM rats.

METHODSD rats were fed high-fat diet for 4 weeks and then were injected STZ (30 mg x kg(-1)) to induce the type 2 diabetes mellitus (T2DM). Once the T2DM models were set successfully, rats were randomized into six groups: normal group (NG), group of diabetes mellitus (DMG), groups of low dosage (GLPs-LG), middle dosage (GLPs-MG), high dosage (GLPs-HG) and berberine (BerG). They received GLPs with different dosages (200, 400, 800 mg x kg(-1)) and berberine (30 mg x kg(-1)) continually for 16 weeks. At 16th weekend, the following indices of rats were measured respectively: blood glucose, hemodynamic including LVSP, LVEDP, dp/dt(max) and -dp/dt(max) and the contents of NO, SOD, MDA, GSH-Px, CAT in cardiac tissue. Besides, myocardial ultrastructure was observed by electron microscope.

RESULTBoth the middle dosage and the high dosage of GLPs could low blood glucose effectively, and they could reduce LVEP but increase -dp/dt(max). Meanwhile, they could activate GSH-Px, CAT, SOD, NO, but reduce MDA in cardiac tissue and improve the myocardial ultrastructure. Compared to the DM group, the middle dosage, high dosage of GLPs and berberine showed significant improvement. Compared to the berberine group, the middle dosage showed the same effect, but the high dosage was more effective than berberine.

CONCLUSIONThere is a confirmed action of GLPs in improving the hemodynamic and antioxidation in cardiac tissue of T2DM rats.

Animals ; Antioxidants ; metabolism ; Catalase ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Female ; Glutathione ; metabolism ; Hemodynamics ; drug effects ; Hypoglycemic Agents ; chemistry ; therapeutic use ; Male ; Malondialdehyde ; metabolism ; Microscopy, Electron, Transmission ; Myocardium ; metabolism ; ultrastructure ; Nitric Oxide ; metabolism ; Polysaccharides ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Reishi ; chemistry ; Superoxide Dismutase ; metabolism

By using single spleen colony transplantation technique and sex chromosome typing as natural cytogenetic markers, we have been able to demonstrate that most spleen colony forming cells in the adult bone marrow or in foetal liver of inbred LACA or C57 mice possess the potential of re-establishment of myeloid and lymphoid lines of cells in lethally irradiated mice. However, most of the spleen colony forming cells in the peripheral [blood of normal mice possess weak or no potentiality to proliferate or re-establish hemopoiesis in lethally irradiated mice. Our results support the notion that the CFU-S population in mice is heterogeneous.From the nature of colony forming cells in the peripheral blood of normal mice, we are convinced that the assessment of CFU-S content relying upon the spleen colonies as the unique sign to indicate the self-renewal ability of a spleen colony forming cell is by no means valid inasmuch as some plu-ripotent . hemopoietic progenitors derived from pluripotent hemopoietic stem cells may have the same ability to form colonies in vivo.