This study is to identify Chinese medicinal materials Rhizoma et Radix Heraclei, Angelicae Sinensis, Radix Angelicae Pubescentis and Rhizoma et Radix Notopterygii based on ITS2 and its secondary structure. Total 26 ITS sequences of 7 species were downloaded from GenBank, the ITS2 sequences were annotated by HMMer method. The NJ phylogenetic tree was built by MEGA software, the intraspecific and interspecific K2P genetic distance were analyzed by MEGA as well. The ITS2 secondary structures of all taxa were predicted by ITS2 database. Sequence matrix of primary structure and secondary structure was aligned by 4Sale software. And the profile neighbor joining (PNJ) phylogenetic tree was constructed via the the ProfDistS software based on the distance method. The results show that, the average interspecific genetic distance was far greater than the average intraspecific genetic distance, an obvious barcoding gap was noted among all taxa; NJ tree showed that all species were clustered into seperate branches; each species had different secondary structures; the PNJ tree showed higher resolution than NJ tree. Therefore, ITS2 is suggested to be used as a barcode for distinguishing the original plants of Rhizoma et Radix Heraclei, Radix Angelicae Sinensis, Radix Angelicae Pubescentis and Rhizoma et Radix Notopterygii in this study, this provides some scientific basis for classification and accurate identification of these Chinese medicinal materials.

OBJECTIVETo explore an approach to rapidly and accurately identify the compounds as biomarkers of Chinese medicine (CM) syndromes.

METHODThe Fourier transform infrared (FT-IR) spectrometry was applied to investigate the characteristic components of a mice model of Kidney (Shen)-yang deficiency syndrome (KDS), and the remedial effect of a typical CM formula Shenqi Pill (). Thirty-six females and 18 males of Balb/c mice were randomly divided into KDS, Shenqi or control group. The females and males of the same group freely were mated for 96 h, and the males were taken out and only the female mice were raised. Females of the KDS group were threatened by a ferocious cat every other day for 14 d. After delivery, the KDS, or gestational threatened, offspring were raised at standard condition for 11 weeks. Then 10 male offspring were randomly selected, anaesthetized and their representative organs, i.e. testes, kidneys, lungs and feet were collected, for the FT-IR scan. Mice of the Shenqi group were intragastric administered Shenqi Pill; while mice in the KDS and control groups were given the same volume of saline.

RESULTSThe attenuated birth outcomes of the KDS group were displayed. The remarkable FT-IR differences of all organs between KDS mice and healthy control were mainly at 1,735-1,745 cm(-1) (indicating the increased levels of lipids) and at 1,640-1,647 cm(-1) and 1,539-1,544 cm(-1) (displaying the decreased proteins). No statistic FT-IR difference between Shenqi and control mice was observed.

CONCLUSIONIn accordance with major traits of KDS, prenatal stress extensively impaired the building up of proteins and resulting in the excessive lipid storage, and FT-IR could effectively identify the biomarkers of KDS.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Kidney Diseases ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Spectroscopy, Fourier Transform Infrared ; methods ; Yang Deficiency ; drug therapy ; pathology

The objective of study was to explore the influence of high mobility group box 1 (HMGB1) on migration of cord blood CD34(+) cells and their mechanism of migration. The expressions of receptor for advanced glycation end products (RAGE), toll-like receptor-2 (TLR2) and TLR4 were detected by flow cytometry. The CD34(+) cells in umbilical cord blood (CB) were enriched by MiniMACS and were exposed to various concentration of HMGB1 (10, 50, 100, 1, 000 ng/ml), then the migration effect of HMGB1 on umbilical cord blood (UCB) CD34(+) cell count was determined by microscopy, the chemotactic index was calculated. The CD34(+) cells untreated with HMGB1 were used as control. The results indicated that the purity of the isolated CD34(+) cells was more than 98%. The HMGB1 could promote the migration of CD34(+) cells, and the migration effect of HMGB1 on CD34(+) cells in certain concentrations gradually increased along with raise of concentration, the strongest effect was observed in concentration of 100 ng/ml, there was significant difference as compared with control (p < 0.01). Anti-RAGE antibody partially inhibited the migration effect of HMGB1 on CD34(+) cells. It is concluded that the HMGB1 in certain concentration can enhance migration of CD34(+) cells, which may be mediated through RAGE.
Antigens, CD34 ; Cell Movement ; drug effects ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; drug effects ; HMGB1 Protein ; pharmacology ; Humans ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Signal Transduction

Two human Fab antibodies against avian influenza A (H5N1) virus were obtained by panning a H5N1 patient-derived antibody phage library using purified virions of the H5N1 patient isolate A/Anhui/1/2005 and HA protein of the H5N1 reference viruse A/Viet Nam/1203/2004. After testing the binding properties and antiviral function to H5N1 virus, the selected Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell system. Both mAbs, AVFluIgG01 and AVFluIgG03, bound to HA in immunofluorescence assay (IFA) without cross-reaction with the other substypes of influenza A viruses (H1N1, H3N2). The cross-reactivity of the two antibodies for different strains of H5N1 was tested in vitro by micro-neutralization assays. In vitro, mAb AVFluIgG01 potently neutralized not only the selected well-characterized Clade 2 H5N1 viruses isolated from mainland of China except A/Guangdong/1/2006, but also the Clade 1 representative isolate A/Viet Nam/1203/2004; and AVFluIgG03 neutralized all the selected Clade 2 H5N1 viruses isolated from mainland of China, but had no neutralizing activity with the Clade 1 H5N1 virus A/Viet Nam/1203/2004. The results bring new prospect for the prophylaxis or treatment of H5N1 virus infection and may provide a clue for novel vaccine development.
Amino Acid Sequence ; Animals ; Antibodies, Viral ; genetics ; immunology ; Antibody Specificity ; Birds ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; genetics ; immunology ; Influenza in Birds ; immunology ; virology ; Molecular Sequence Data ; Neutralization Tests ; Recombinant Proteins ; immunology ; Sequence Homology, Amino Acid