AIM　To study the effects of dexamethasone on expressing monocyte chemoattractant protein-1(MCP-1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS　The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxsip, the levels of MCP-1 mRNA were determined by RT-PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS　The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP-1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION　The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP-1 mRNA.

Objective To compare the expression changes of neuropeptide Y (NPY) receptor Y1 in different stages of osteoblast differentiation of rat bone marrow mesenchymal stem cells (BMSCs).Methods The rBMSCs were isolated in vitro from Sprague-Dawley (SD) rats using whole bone marrow adherence method and cultured. Then, the rBMSCs were divided into osteoblast-induced group and noninduced group. In different periods of culture at 1, 2 and 3 weeks, identification of the osteoblasts was performed by using immunocytochemistry and Western blot. Expressions of mRNA and protein of Y1 receptor were detected by real time reserve transcriptase-polymerase chain reaction (RT-PCR) and Western blot. Results RT-PCR demonstrated that osteoblast-induced group had a lower expression of Y1 receptor than non-induced group at the same time point and the expression of Y1 receptor was increased in a time-dependent manner in both groups. Western blot demonstrated higher expression of Y1 receptor in osteoblast-induced group compared with non-induced group at the same time point and a decreased expression of Y1 receptor in a time-dependent manner in both groups. Conclusions During the process of osteoblastic differentiation of rat BMSCs, the expressions of mRNA and protein of NPY Y1 receptor show different trends, when NPY may mediate the inhibition of osteoblastic differentiation of BMSCs through Y1 receptor pathway.

Objective To investigate the effects and mechanism of calcitonin gene-related peptide (CGRP) and substance P (SP) on proliferation of rat bone marrow mesenchymal stem cells. Methods The rBMSCs were isolated using whole bone marrow adherence method. In the different periods of culturing (1, 2,and 3 weeks), expressions of the neuropeptide receptors were detected by Western Blot and reserve transcriptase-polymerase chain reaction (RT-PCR). The BMSCs were treated with CGRP and SP at concentration 10-8 mol/L at different time (1,3,5,7,9 days), cell proliferation was detected with MTT assay, the protein expressions of cyclin D1 ,cyclin E and p53 were examined using Western Blot. Results The CGRP receptor and SP receptor were expressed in BMSCs. The expression of CGRP receptor was statistically higher than that of SP receptorat the same time point. The growth curves of BMSCs cultured by both neuropeptides had similar appearance. CGRP and SP stimulated the proliferation of BMSCs significantly at 9 days and 7 and 9 days. In this process, the expressions of cyclinDl and cyclinE were up-regulated by CGRP, SP only enhanced the expression of cyclinE; these effects all reached a peak at 5 days. The expression of p53 was down-regulated by both neuropeptides. Conclusion CGRP and SP had direct effects on the proliferation of BMSCs, the regulation of cell cycle proteins is one of the mechanisms.

To analyze the composition regularity of Carthami Flos-containing prescriptions of the Drug Standards of Ministry of Health of People's Republic of China-Traditional Chinese Medicine Preparations (the ministerial standards for Traditional Chinese Medicine) based on the traditional Chinese medicine inheritance support system (TCMISS, RZDZ No. 0389952). Efforts were made to identify 331 prescriptions containing Carthami Flos and summarize 16 attending functions and 10 commonly used drug combinations. Three commonly used drug combinations were selected for an in-depth analysis on Carthami Flos's combined administration regularity. Based on Carthami Flos's attending functions, its effects in paralysis, traumatic injuries and dysmenorrheal were compared to analyze Carthami Flos's core drug combinations for treating different diseases. The regularity of clinical administration and the characteristics of commonly used drug combinations were summarized to provide reference for Carthami Flos's clinical application and new ideas for new drug R&D. Carthami Flos prescriptions was mainly used to treat blood stasis and pain and mostly combined with drugs that could activate blood, promote the circulation of qi and dispel pathogenic wind to treat Qi-stagnation and blood stasis caused by various pathogenic factors such as wind, cold and dampness.
Carthamus ; chemistry ; Clinical Trials as Topic ; Drug Prescriptions ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Flowers ; chemistry ; Humans

AIM To study the effects of dexamethasone on expressing monocyte chemoattractant protein 1(MCP 1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxs ip , the levels of MCP 1 mRNA were determined by RT PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP 1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP 1 mRNA.

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.
Caffeic Acids ; analysis ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Hydroxybenzoates ; analysis ; Lonicera ; chemistry ; Quinic Acid ; analogs & derivatives ; analysis

ObjectiveTo study the effect of fluorine on proliferation of osteoblast through extra cellular signal-regulated protein kinase(ERK) signaling pathway.MethodsMouse osteoblasts(MC3T3-E1) were cultured in vitro with different concentrations of fluoride for 24 and 48 h (the concentrations of Fˉ were 0,200,400,600,1000,2000,4000,8000,10 000 μmol/L,respectively).The optimum concentration for promotion of cell proliferation was determined by methylthiophene tetrazolium(MTT) assay.According to the optimum concentration,the cells were randomly divided into three groups:control group (0 μmol/L Fˉ); fluorine group (400 μmol/L Fˉ); fluorine and MAPK inhibitor PD98059 group(400 μ mol/L Fˉ + 10 μ mmol/L PD98059).Cell cycle was detected by flow cytometry after 48 h culture.The expression of P-ERK protein was determined by Western blotting and immunofluorescence.ResultsThe optimum concentration of fluorine for proliferation of osteoblasts was 400 μ mol/L.Compared with the control group[(76.12 ± 10.08)％,(2.06 ± 0.31)％],the number of cells in G0/G1 phase[(63.04 ± 8.12)％] reduced and the number of cells in S phase[(9.13 ± 2.08)％] increased in fluorine group (all P ＜ 0.05) ; but the number of cells in G0/G1 phase [(92.11 ± 9.01 ) ％] in fluorine and mitogen-activated protein kinases (MAPK) inhibitor PD98059 group was significantly increased(P ＜ 0.05 ).Western blotting results showed that:compared with the control group[(100.00 ± 0.00)％],the expression of P-ERK protein in fluorine group[(131.24 ± 13.88)％] was significantly higher(P ＜ 0.05 ),but the expression of P-ERK protein in fluorine and MAPK inhibitor PD98059 group [(91.33 ± 9.68 )％] was not significantly changed(P ＞ 0.05).The results of immunofluorescence were similar to that of Western blotting.ConclusionsFluorine at the concentration of 400 μmol/L can promote the proliferation of osteoblasts.ERK signaling pathway has played a key role in the proliferation of osteoblasts.

Objective To establish a new method for measuring the 3-D kinematics of hindfoot joints in vivo by using reverse engineering software together with the theory of rigid body kinematics.Methods CT images were gathered from 9 feet of 5 healthy volunteers in both the initial position (neutral position) and the terminal position (extremely inversion-adduction-dorsiflexion position).The 3-D digital modules of hindfoot joints in the initial position and terminal position were established with MIMICS 10.01 software.The data of reconstructed digital modules was inputted into the GEOMAGIC 10.0 software in STL format for twice registration,and then their relatively displacement and changes of angle in 3-D space between the two positions were calculated Results The rotation range of the tibiotalar joint was 3.89° ±2.77° in eversion,5.29°±4.47° in abduction,10.77°+5.70° in dorsiflexion,and the relative displacement was 0.78±0.59 mm towards lateral ankle,0.18±0.75 mm towards the hindfoot,(0.65±0.71) mm towards the proximal limbs;the range of subtalar joint was 16.46°±2.94° in inversion,12.77°±1.81° in adduction,6.33°±4.32° in plantarflexion,and the relative displacement was 5.50±1.45 mm towards medial ankle,1.96±1.77 mm towards forefoot,0.43±1.18 mm towards distal limbs; the range of talonavicular joint was 38.82°±5.98° in inversion,19.71°±6.33° in adduction,5.09°±6.89° in plantarflexion,and the relative displacement was (9.77±1.73) mm towards medial ankle,3.13±1.29 mm towards hindfoot,4.64±1.42 mm towards proximal limbs.Conclusion This method measuring 3-D kinematics of hindfoot joints in vivo is non-invasive and easy to operate.

Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; mortality ; Humans ; Infant ; Male

This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.
Animals ; Capsid Proteins ; genetics ; Child, Preschool ; China ; epidemiology ; Evolution, Molecular ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; veterinary ; virology ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Nonstructural Proteins ; genetics