Chinese materia medica resource (CMM resource) is the foundation of the development of traditional Chinese medicine. In the study of sustainable utilization of CMM resource, adopting innovative theory and method to find new CMM resource is one of hotspots and always highlighted. Pharmacophylogeny interrogates the phylogenetic relationship of medicinal organisms (especially medicinal plants), as well as the intrinsic correlation of morphological taxonomy, molecular phylogeny, chemical constituents, and therapeutic efficacy (ethnopharmacology and pharmacological activity). This new discipline may have the power to change the way we utilize medicinal plant resources and develop plant-based drugs. Phylogenomics is the crossing of evolutionary biology and genomics, in which genome data are utilized for evolutionary reconstructions. Phylogenomics can be integrated into the flow chart of drug discovery and development, and extends the field of pharmacophylogeny at the omic level, thus the concept of pharmacophylogenomics could be redefined in the context of plant pharmaceutical resources. This contribution gives a brief discourse of knowledge pedigree of pharmacophylogeny, epistemology and paradigm shift, highlighting the theoretical and practical values of pharmacophylogenomics. Many medicinally important tribes and genera, such as Clematis, Pulsatilla, Anemone, Cimicifugeae, Nigella, Delphinieae, Adonideae, Aquilegia, Thalictrum, and Coptis, belong to Ranunculaceae family. Compared to other plant families, Ranunculaceae has the most species that are recorded in China Pharmacopoeia (CP) 2010. However, many Ranunculaceae species, e. g., those that are closely related to CP species, as well as those endemic to China, have not been investigated in depth, and their phylogenetic relationship and potential in medicinal use remain elusive. As such, it is proposed to select Ranunculaceae to exemplify the utility of pharmacophylogenomics and to elaborate the new concept empirically. It is argued that phylogenetic and evolutionary relationship of medicinally important tribes and genera within Ranunculaceae could be elucidated at the genomic, transcriptomic, and metabolomic levels, from which the intrinsic correlation between medicinal plant genotype and metabolic phenotype, and between genetic diversity and chemodivesity of closely related taxa, could be revealed. This proof-of-concept study regards pharmacophylogenomics as the updated version of pharmacophylogeny and would enrich the intension and spread the extension of pharmacophylogeny. The interdisciplinary knowledge and techniques will be integrated in the proposed study to promote development of CMM resource discipline and to boost sustainable development of Chinese medicinal plant resources.
China ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Knowledge ; Medicine, Chinese Traditional ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

BACKGROUND:Pedicle screw is the major instrumentation of surgery in thoracic spine. However, there have been few reports about pedicle morphology relevant to screw insertion tracts, and few reports comparing the normal adolescents and adolescent idiopathic scoliosis patients. OBJECTIVE:To compare the morphologic characteristics of the thoracic pedicle with regard to safe thoracic pedicle screw placement in normal adolescents and adolescent idiopathic scoliosis patients. METHODS: Thoracic pedicles of thirty-five normal adolescents and thirty-five adolescent idiopathic scoliosis patients were measured with three-dimensional reconstruction CT images. Measured parameters include (1) critical distance: the shortest distance from an entry point to the ventral cortex of the lamina. (2) Safe distance: the distance from the entry point to the tangent of the spinal canal at the medial wal of the pedicle. (3) Pedicle screw length. (4) Pedicle width. (5) Pedicle transverse angle. The dangerous area was defined as the distance between the critical distance and the safe distance. RESULTS AND CONCLUSION: The mean critical distance was (9.2±1.0) mm for the normal adolescents, and (9.4±1.2) mm for the adolescent idiopathic scoliosis patients. Safe distances were significantly less in normal adolescents (14.7±0.8) mm than that of the adolescent idiopathic scoliosis group (15.4±1.4) mm (P < 0.001). The dangerous area was (5.4±0.7) mm for the normal adolescents, which was significantly less than that of the adolescent idiopathic scoliosis patients (6.0±1.0) mm (P < 0.001). Pedicle screw length was (36.6±4.1) mm for the normal adolescents and (37.1±5.3) mm for the adolescent idiopathic scoliosis patients. Pedicle width was (5.8±1.2) mm for the normal adolescents and (5.7±1.7) mm for the adolescent idiopathic scoliosis patients. No significant difference in critical distance, pedicle screw length and pedicle width was found between the two groups (P=0.382, 0.135, 0.293). Pedicle transverse angle decreased gradualy from T1 to T12 in both groups. These results verify that pedicle morphology of many parameters is different between normal adolescents and adolescent idiopathic scoliosis patients, especialy in the apical area of the thoracic curve.

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

ObjectiveTo investigate the loss of heterozygosity (LOH) on 6q in bladder tumor.MethodsD6S404 and D6434 microsatellite markers near 6q21 were tested by PCR-SSLP-stain method on tumor DNA from 31 cases of bladder tumor.ResultsAmong these 31 cases of bladder tumor,LOH was detected in tumor tissues on site for D6S404 (35.5%) and D6S434(22.6%).ConclusionOne or more tumor suppressor gene near 6q21 maybe relevant for the development of bladder tumor.

With the surge of high-throughput sequencing technology, it is becoming popular to perform the phylogenetic study based on genomic data. A bundle of new terms is emerging, such as phylogenomics, pharmacophylogenomics and phylotranscriptomics, which are somewhat overlapping with pharmaphylogeny. Phylogenomics is the crossing of evolutionary biology and genomics, in which genome data are utilized for evolutionary reconstructions. Pharmaphylogeny, advocated by Prof. Pei-gen Xiao since 1980s, focuses on the phylogenetic relationship of medicinal plants and is thus nurtured by molecular phylogeny, chemotaxonomy and bioactivity studies. Phylogenomics can be integrated into the flow chart of drug discovery and development, and extend the field of pharmaphylogeny at the omic level, thus the concept of pharmacophylogenomics could be redefined. This review gives a brief analysis of the association and the distinguished feature of the pharmaphylogeny related terms, in the context of plant-based drug discovery and sustainable utilization of pharmaceutical resource.
Drug Discovery ; Pharmacogenetics ; Phylogeny ; Plants, Medicinal ; chemistry ; genetics

Adolescent ; Adult ; Aged ; Brain ; blood supply ; diagnostic imaging ; Carbon Monoxide Poisoning ; diagnostic imaging ; Child ; Female ; Humans ; Male ; Middle Aged ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo evaluate the effect of hypoxia-inducible factor-1 alpha (HIF-la) on angiogenesis in human prostate cancer cells.

METHODSHuman prostate cancer cells of the line LNCaP were cultured and transfected by the recombinant plasmid pcDNA3. 1(-)/HIF-1alpha containing the gene HIF-1alpha with the Lipofectamine 2000 system. The positive clone cells were selected by G418 and confirmed by Western blotting and immunofluorescence (LNCaP/HIF-1alpha cells). The expressions of VEGF, iNOS and Ang-2 were detected by Western blotting.

RESULTSThe expression of HIF-1alpha in the LNCaP/HIF-1alpha cells was distinctly higher than that in the LN-CaP cells. Compared with the LNCaP cells, the expressions of VEGF and iNOS were up-regulated, whereas Ang-2 remained unchanged in the LNCaP/HIF-1alpha cells.

CONCLUSIONThe over-expression of HIF-1alpha can induce an increase in angiogenesis proteins and improve the angiogenesis potency of prostate cancer.

Blotting, Western ; Cell Line, Tumor ; Fluorescent Antibody Technique ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Lipids ; chemistry ; Male ; Nitric Oxide Synthase ; metabolism ; Plasmids ; chemistry ; genetics ; Prostatic Neoplasms ; blood supply ; metabolism ; pathology ; Transfection ; methods ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effects of the over-expression of hypoxia-inducible factor-1alpha (HIF-1alpha) on the epithelial-mesenchymal transition (EMT) and invasive potency of LNCaP cells in vitro and in vivo.

METHODSWe cultured LNCaP cells stably expressing HIF-1alpha (LNCaP/HIF-1alpha) and LNCaP cells, identified the over-expression of HIF-1alpha, determined the proliferation of the two cell lines by MTT assay and the level of PSA in the supernatant of culture medium, and detected the anchorage independent growth by soft-agar colony formation assay. A subcutaneous tumor model was established in nude mice by injecting LNCaP/HIF-1alpha and LNCaP cells followed by observation of the tumor growth. Tumor specimens were obtained for immunohistochemistry.

RESULTSThe over-expression of HIF-1alpha was confirmed in the LNCaP/HIF-1alpha cells by immunofluorescence staining and Western blotting. The level of PSA was obviously decreased in LNCaP/HIF-1alpha as compared with that in LNCaP cells. MTT assay identified the increased proliferation of LNCaP/HIF-1alpha cells. The cell colony forming ability of the LNCaP cells was significantly lower than that of the LNCaP/HIF-1alpha cells. The rate of tumorigenesis was increased and its time shortened in the LNCaP/HIF-1alpha group. Immunohistochemistry revealed an up-regulated expression of vimentin and a down-regulated expression of E-cadherin in the tumor specimens.

CONCLUSIONThe overexpression of HIF-1alpha can up-regulate the expression of vimentin and down-regulate the expression of E-cadherin, which may enhance the invasive potency of LNCaP cells by inducing EMT.

Animals ; Cell Hypoxia ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; metabolism ; pathology

OBJECTIVETo study the effects of Liangge San to the expression of CD14 and scaverger receptor(SR) in the kupffer cells of liver and the pathological changes of liver tissue of endotoxemia-mice.

METHODThe model was established with intravenous injection of lipopolysaccharide (LPS) and at the same time different dose Liangge San were given. The expression of CD14 and scaverger receptor were detected with immunohigtochemistry at the 2nd, 4th, 8th hour ofter injury and analyzed with computer image system, and the pathological changes of liver tissue were also observed.

RESULTAt the three different hours, the expression of CD14 and scaverger receptor in macrophages of liver of LPS-injury group showed significant increase and significant decrease respectively, compared with that of the blank-control group (P < 0.01). The expression in dexamethasone group and Liangge San different dose groups were intermediate between those in injury group and those in control group. Compared with expression of LPS-injury group, those of dexamethasone group and Liangge San different dose groups showed significant differences (P < 0.01), especially that of Liangge San high dose group. Liver cells showed vacuole change. Changes of CD14 and SR expression were paralleled with the severity of liver damages of the mice.

CONCLUSIONLiangge San can inhibite the up-regulation of CD14 expression and down-regulation of scaverger receptor expression in a dosage-dependent manner and also alleviate the damages of liver induced by LPS.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endotoxemia ; chemically induced ; metabolism ; pathology ; Female ; Kupffer Cells ; metabolism ; pathology ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; Liver ; metabolism ; pathology ; Mice ; Plants, Medicinal ; chemistry ; Receptors, Scavenger ; metabolism

OBJECTIVETo study the regulatory effect of Qinghuo Baidu Yin (QHBDY, a mixture prepared with Chinese drugs) on immune function of patients with extremely severe burn (ESB).

METHODSThirty patients with ESB were divided into two groups, conventional therapy was given to both groups, but QHBDY was given to the treated group additionally. Immunological indices, including peripheral blood T-lymphocyte subsets, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) were determined 3 weeks after treatment to evaluate and compare the therapeutical effect in the two groups.

RESULTSCompared with the control group, CD3, CD4, CD4/CD8, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) levels were markedly decreased in degree, and recovered earlier and quicker, with CD8 increased mildly (P < 0.01) and turned back more quickly. And so did the parameters of the treated group in comparing with that of the control group at anytime (P < 0.05 or P < 0.01).

CONCLUSIONChinese drugs mixture shows the regulatory effect on both cellular and humoral immune function in patients with ESB.

Adjuvants, Immunologic ; therapeutic use ; Adolescent ; Adult ; Burns ; drug therapy ; immunology ; Complement C3 ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Phytotherapy