Exploring Anti-inflammation Mechanism of Pien Tze Huang via Regulating of Microglia Polarization / 中国实验方剂学杂志

Objective::To investigate the anti-inflammation mechanism of Pien Tze Huang (PTH) via regulating microglia polarization. Method::The experiment was divided into five groups, Blank, M1[lipopolysaccharide (LPS) 100 μg·L-1+ interferon-γ(IFN-γ) 10 μg·L-1], M1-PTH group[LPS 100 μg·L-1+ IFN-γ 10 μg·L-1+ PTH 0.4 g·kg-1], M2 group[interleukin-4 (IL-4) 20 μg·L-1], and M2-PTH group[IL-4 20 μg·L-1+ PTH 0.4 g·kg-1]. The concentration of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and transforming growth factor-β1(TGF-β1) in the culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA), the levels of inducible nitric oxide synthase(iNOS) and arginine-1 (Arg-1) mRNA were detected by real-time fluorescent quantitative polymerase chain reaction technique(Real-time PCR), and the expression levels of p-STAT1, p-STAT3, iNOS, p-STAT6, and Arg-1 were detected by Western blot. Result::The concentration of NO and TNF-α of the culture supernatant, the level of iNOS mRNA, as well as the level of p-STAT1, p-STAT3 and iNOS in M1 group, which were significantly increased(P<0.01) .Compared with blank group, but the concentration of NO and TNF-α were down-regulated(P<0.01), and iNOS mRNA(P<0.05), as well as the expression of iNOS, p-STAT1, and p-STAT3 was decreased (P<0.05, P<0.01) after the invention of PTH in M1-PTH group compared with M1 group. The concentration of IL-10 and TGF-β1 in the culture supernatant, the mRNA level of Arg-1, as well as the levels of p-STAT6 and Arg-1 were significantly increased in M2 group when compared with Blank group, addition to the concentration of IL-10 and TGF-β1 were up-regulated(P<0.05, P<0.01), and the expression of Arg-1 mRNA, the level of Arg-1, p-STAT6 were enhanced(P<0.05, P<0.01) in M2-PTH group compared with M2 group. Conclusion::PTH plays an anti-inflammatory role via regulating microglia polarization.

Systems-Based Interactome Analysis for Hematopoiesis Effect of Angelicae sinensis Radix: Regulated Network of Cell Proliferation towards Hemopoiesis / 中国结合医学杂志

OBJECTIVE@#To explore the molecular-level mechanism on the hematopoiesis effect of Angelicae sinensis Radix (ASR) with systems-based interactome analysis.@*METHODS@#This systems-based interactome analysis was designed to enforce the workflow of "ASR (herb)→compound→target protein→internal protein actions→ending regulated protein for hematopoiesis". This workflow was deployed with restrictions on regulated proteins expresses in bone marrow and anemia disease and futher validated with experiments.@*RESULTS@#The hematopoiesis mechanism of ASR might be accomplished through regulating pathways of cell proliferation towards hemopoiesis with cross-talking agents of spleen tyrosine kinase (SYK), Janus kinase 2 (JAK2), and interleukin-2-inducible T-cell kinase (ITK). The hematopoietic function of ASR was also validated by colony-forming assay performed on mice bone marrow cells. As a result, SYK, JAK2 and ITK were activated.@*CONCLUSION@#This study provides a new approach to systematically study and predict the therapeutic mechanism for ASR based on interactome analysis towards biological process with experimental validations.

Side Effect and Attenuation of Triptolide / 中国实验方剂学杂志

Triptolide (TP) is a kind of epoxy diterpene lactone compound extracted from xylem of Tripterygium wilfordii (TWHF). It is one of the main active components of TWHF, with anti-inflammatory, immunomodulatory and anti-tumor activities and pharmacological effect. It can be used in the treatment of rheumatoid arthritis and such cancers as leukemia, breast cancer, pancreatic cancer and lung cancer. In recent years, TP has been gradually applied in clinic and basic research, with a certain curative effect. But it also has certain side effect on digestive system, urinary system, reproductive system, circulatory system and immune system, including liver dysfunction, reduced fertility, chest tightness, heart palpitations, bradycardia, atrioventricular block, arrhythmia, kidney dysfunction, lymphoid organ atrophy, lymphoid tissue necrosis of lymphocytes, and reduction in the number of cells, impaired immune function. These toxic and side effect have greatly restricted the clinical application of TP. In the meantime, the researches and development related to preparations of TP have also been restricted, which has aroused wide attention from clinicians and researchers. To improve the therapeutic effect of TP and reduce the toxicity of TP in the process of application, domestic and foreign researchers have made a lot of studies and attempts, such as changing the chemical structure of TP to improve its solubility, developing drug delivery system to reduce its toxicity and using combination therapy with traditional Chinese herbal medicines to increase efficiency and reduce toxicity. In this paper, the toxic dose and mechanism of TP, TP's derivatives, drug loading system, compatibility and attenuation were integrated to provide ideas for further researches on toxicity and attenuation of TP.

Bioinformatics analysis on immune response pathways of triptolide in treating AIDS / 中国中药杂志

The aim of this paper was to study the influence of triptolide in the immune response pathways of acquired immune deficiency syndrome( AIDS). Target proteins of triptolide and related genes of AIDS were searched in PubChem and Gene databases on line. Molecular networks and canonical pathways comparison analyses were performed by bioinformatics software( IPA). There were 15 targets proteins of triptolide and 258 related genes of AIDS. Close biological relationships of molecules of triptolide and AIDS were established by networks analysis. There were 21 common immune response pathways of triptolide and AIDS,including neuroinflammation signaling pathway,Th1 and Th2 activation pathway and role of pattern recognition receptors in recognition of bacteria and viruses. Triptolide stimulated immune response pathways by the main molecules of IFNγ,JAK2,NOD1,PTGS2,RORC. IFNγ is the focus nodes of triptolide and AIDS,and regulates genes of AIDS directly or indirectly. Triptolide may against AIDS by regulating molecules IFNγ in immune response pathways.

Effect of Anerdian Ⅲ Disinfectant On the Function of Eye Surface and Tear Film / 中山大学学报(医学科学版)

@#【Objective】To evaluate the effect of preoperative conjunctival sac irrigation with Anerdian Ⅲ disinfectant and its effect on the function of eye surface and tear film. 【Methods】 This clinical study was involved of thirty cases （30 eyes） undergoing phacoemulsification. Conjunctiva sac irrigation with Anerdian disinfectant （type III ，5g/L） was performed before the phacoemulsification procedure. Secretions of conjunctiva sac were examined by bacterial culture pre-and post-irrigation. Schirmer test，non-invasive first tear break-up time（NITBUT-F），non-invasive average tear break-up time（NITBUT-A），tear meniscus height（TMH），and goblet cell density（GCD）were also evaluated day 1 before and day 7 ，day 30 after operation.【Results】3 cases were positive in bacterial culture pre-irrigation ，while all were negative post-irrigation. There was no significantly change between Schirmer test and TMH from day 1 before ，day 7 and day 30 after operation . NITBUT-F，NITBUT-A，and GCD were significantly different between day 1 before and day 7 after operation（P<0.05），and shown no significantly difference between day 1 before and day 30 after operation.【Conclusions】 Our results suggested that conjunctiva sac irrigation with Anerdian disinfectant was efficient，while it might be related to the decrease of GCD and dysfunction of tear film. However，GCD would gradually improve as pre-operation and function of tear film could restore to normal.

Protective Effect of Norcantharidin on Collagen-Induced Arthritis Rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of norcantharidin (NCTD) on collagen-induced arthritis (CIA) rats.</p><p><b>METHODS</b>Sixty Sprague-Dawley(SD) rats were randomly divided into 6 groups (n=10): normal group, CIA model group(model group), NCTD low-dose group [1.35 mg/(kg•d)], NCTD middle-dose group [2.7 mg/(kg•d)], NCTD high-dose group [5.4 mg/(kg•d)] and methotrexate (MTX) group [1.8 mg/(kg/w)]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin (H&E) staining. The serum levels of interleukin (IL) 1β, IL-6, tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), IL-17 and transform growth factor (TGF) β were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression of retinoid-related orphan nuclear receptorγt (RORγt) and forkhead box P3 (Foxp3) in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction.</p><p><b>RESULTS</b>MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats' ankle joints compared with the model group (P<0.05 or P<0.01). All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats (P<0.05). Only middle- and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats (P<0.05). However, NCTD has no effect on vascular endothelial growth factor (VEGF) level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group (P<0.05). The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group (P<0.05).</p><p><b>CONCLUSIONS</b>NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.</p>

Explore pharmacological mechanism of glycyrrhizin based on systems pharmacology / 中国中药杂志

To explore the pharmacological mechanism of glycyrrhizin with series methods of systems pharmacology, main diseases related to glycyrrhizin were obtained by text mining tool; and the target proteins of glycyrrhizin were obtained via the database of Polysearch and PubChem. Then, the target proteins interaction network of glycyrrhizin was built using the software called Cytoscape. Next, the protein groups related to glycyrrhizin were analyzed by using Gene Ontology (GO) tool, and the action pathway of its target proteins was analyzed by using enrichment method. Text mining results showed that the related diseases of glycyrrhizin included chronic hepatitis C, chronic hepatitis, hepatitis, HIV virus, liver cancer and so on. Gene ontology analysis indicated that glycyrrhizin played a role mainly through modification of proteins and chromatin. The signaling pathway enrichment results showed that the main action proteins of glycyrrhizin were related to MAPK signaling pathway, toll-like receptor signaling pathway, neurotrophic factor signaling pathway, cancer and apoptosis pathways. So we can conclude that glycyrrhizin may exert its biological functions primarily by regulating multiple pathways such as MAPK signaling pathway and Toll-like receptors signaling pathway. The pharmacological action of a drug can be rapidly and comprehensively analyzed by the ways of systems pharmacology.

Clinical trials of integrative medicine for rheumatoid arthritis: Issues and recommendations / 中国结合医学杂志

Controlled clinical trials of integrative therapies available to patients with rheumatoid arthritis (RA) improved dramatically in the past 20 years, largely because of the growing need and the methodologies improvement. Tripterygium wilfordii Hook. F., a typical example of popular use herb, has been extensively studied in trials. However, clear and convincing evidence of integrative therapy, effectiveness and safety, remains insufficient to make decision. Many research efforts are hampered by standing problems with 'syndrome' recruitment failure. In addition, the outcome multiplicity induces the findings inefficiency to generalize to RA patients at large. Development of validated syndrome outcomes and methodologies has also been critical. Current efforts to enhance the understanding of integrative treatment options for patients with RA include the development of drug-specific rather than disease-specific strategies, studies in predictive biomarkers, and development of peer-review trial protocol for regular clinical trials.

Effect of Combination Therapy of Tetramethylpyrazine with Methotrexate on Inflammatory Reac- tions and Hemorheology in Collagen-induced Arthritis Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of combination therapy of tetramethylpyrazine (TMP) with methotrexate (MTX) on collagen induced arthritis (CIA) rats.</p><p><b>METHODS</b>Totally 55 male SD rats were stratified by body weight. Nine of them were randomly recruited as the normal control group. The rest 46 were immunized with type II bovine collagen (C II) for establishing rheumatoid arthritis (RA) model. Forty successfully modeled rats were randomly divided into 4 groups according to swollen toe degree, i.e., the CIA group, the TMP group, the MTX group, and the TMP plus MTX group, 10 in each group. Rats in the MTX group were administered with MTX (1. 2 mg/kg) , once per week for 4 continuous weeks. Those in the TMP group were administered with 40 mg/kg TMP, once per day for 10 continuous days, and then discontinued for 7 successive days, and continued for another 10 successive days. Rats in the TMP plus MTX group were administered with a mixture of equal dose MTX and TMP, and when MTX was discontinue, TMP was administered according to the way in the TMP group. Equal volume of saline solution was given to rats in the normal control group and the CIA group. Clinical parameters including ankle width (mediolateral diameter) and hindpaw swelling were measured at day 0, 4, 11, 18, and 26 after treatment. Rats were sacrificed 28 days after treatment, their knee joints and ankle joints were collected for pathological analyses. Serum levels of IL-1β, IL-6, and IL-17A were detected by ELISA. Changes of fibrinogen (FIB) and platelet aggregation rate (PAg) were detected.</p><p><b>RESULTS</b>Compared with the normal control group, the ankle width and hindpaw swelling increased significantly (P < 0.01), contents of FIB and PAg increased obviously (P < 0.05, P < 0.01), serum levels of IL-1β, IL-6, and IL-17 increased remarkably (P <0. 01) in the CIA group. Obvious cell proliferation, inflammatory cell infiltration, hyperemia and edema of synovial tissues could be seen. Pannus formed and immerged in cartilages, resulting in necrosis. Compared with the model group, changes of ankle width and hindpaw swelling were all alleviated in each medicated group (P <0. 05, P <0. 01). Of them, the effect was superior in the MTX group to that of the TMP group and the MTX plus TMP group (P < 0.05, P < 0.01). Contents of FIB, serum levels of IL-1β and IL-6 decreased significantly in the MTX group (P < 0.05). Contents of FIB, serum levels of IL-1β and IL-6 decreased significantly in the TMP group and the MTX plus TMP group (P < 0.05). Besides, serum levels of FIB and IL-6 were obviously lower in the MTX plus TMP group than in the TMP group and the MTX group (P < 0.01). Levels of PAg and IL-17A were more significantly lowered in the TMP group than in the MTX plus TMP group and the MTX group. Pathological changes could be alleviated in each medicated group, with the optimal effect obtained in the MTX plus TMP group.</p><p><b>CONCLUSION</b>Combination of TMP with MTX could significantly ameliorate inflammatory reactions and FIB contents of CIA rats.</p>

Modern elucidative strategies for scientific connotation of controlling toxic reactions while toxic herbs are used to the indication syndrome / 中国中西医结合杂志

One of effective measures for controlling toxic reactions is to use toxic herbs according to corresponding indication syndrome. It is important to develop toxicity theory of Chinese medicine in a sound and international way using modern language to elucidate its scientific connotation. We expect to explain scientific connotation of controlling toxic reaction while toxic herbs are used to the indication syndrome by using holistic research ideas and methods capable of reflecting governing exterior to infer interior, establish appropriate corresponding syndrome animal models by cutting into dose-effect/toxicity of toxic Chinese herbs, construct and analyze multi-layer molecular network using theories and technologies of metabonomics, network biology, and bioinformatics.

Construction of eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment gene G22 and its expression / 中南大学学报(医学版)

OBJECTIVE@#To construct a eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment (ScFv) gene G22, and to identify its expression in rectal cancer cells (CMT-93).@*METHODS@#The G22 gene was ligated into the sites of EcoRI and NotI of eukaryotic expression vector pcDNA3.1(+). After the identification and DNA sequencing, the recombinant plasmid pc DNA3.1(+)-G22 was stably transfected into CMT-93 cells, and the expression of G22 was detected by Western blot, flow cytometry and immunofluorescence staining.@*RESULTS@#Restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid contained the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22. Transfection experiment verified that G22 gene could be expressed in CMT-93 cells in the right way.@*CONCLUSION@#The eukaryotic expression vector containing the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22 is successfully constructed and expressed, which is the basis for further study of its DNA vaccine.

Construction of cDNA library from NPC tissue and screening of antigenic genes / 生物工程学报

To obtain the NPC-associated antigens, a powerful new method, SEREX (serological identification of antigen by recombinant cDNA expression library), was used for identifying the antigens eliciting humoral immune response. Before performing serological analysis, a high quality cDNA library derived from human nasopharyngeal carcinoma (NPC) tissue was constructed. The primary library consisted of 3.64 x 10(6) recombinants and the recombinant rate was 94%. For better preserving the cDNA library, it was amplified. As a result, the titer of the amplified cDNA library was 3.8 x 10(9) pfu/mL. With SEREX method, immunoscreening for the detection of reactive clones in the human NPC tissue cDNA library was performed with autologous serum. As a result, 23 positive clones encoding antigenic genes were obtained after immunoscreening, and the nucleotide sequences of cDNA inserts were determined and analyzed with BLAST software in GenBank. Results showed that the 23 reactive clones were derived from 16 different genes. 10 of 16 genes had high homologous to the genes known in GenBank, such as RPL31, S100 A2, MT2A, etc. However, there were also 6 genes with low homology to the genes known in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be revealed by further research.