This study was aimed to collect relevant provisions of bulging disease medical records in the Ming and Qing dynasties to excavate the syndrome differentiation, drug law and medication frequency analysis. A database was established to analyze relevant provisions. The results showed that the syndromes of liver stagnation and spleen deficiency, dampness-heat accumulation, spleen-yang deficiency are the most common type. The mainly used drugs are from the category to eliminate dampness and water, and to tonify the deficiency. The top three used drugs are poria, atractylodes, and dried citrus peel. It was concluded that medications used in the Ming and Qing dynasties were mainly targeted to the disordered zangfu-organ, which was mainly about the spleen and the stomach. The syn-drome differentiation is the mixture of deficiency and excess. And deficiency is the main part. The medication should combine tonification and reducing. And tonification should be paid attention to. The syndrome differentiation and treatment should be coordinated and the primary and secondary aspect should be identified. These rules provide ref-erence effect for the clinical practice and scientific research of bulging disease treatment.

Objective To discuss the teaching outcomes of problem-based learning method in classroom teaching of TCM pharmaceutics.Methods Fifty students majoring in 2009 science of Chinese materia medica were taught by using traditional teaching method in teaching of TCM pharmaceutics, and 56 students majoring in 2009 TCM pharmaceutics by using of problem-based learning method. This study evaluated students’ adaption to the two teaching methods through score analysis and questionnaire of the two classes.Results Students taught by problem-based learning method generally believed that this teaching method can stimulate their learning interest, improve self-study ability, thinking ability, creativity, and language competence, and enhance team spirit.Conclusion Problem-based learning method shows better teaching outcomes than traditional teaching method. It can guide students to look for ways to tackle problems in the course of solving problems, and cultivate all-round ability and comprehensive quality of students.

Objective · To investigate the effects of insulin on high glucose-cultured humanmononuclear cell line THP-1 and macrophage phenotype transformation in diabetic wounds. Methods · THP-1 cells were cultured with normal (5.6 mmol/L) and high (25 mmol/L) glucose, respectively,stimulated with PMA for differentiation, and induced to M1 macrophages with LPS. After treated with insulin for 6 h, expression changes of M1 type macrophage markers inducible nitric oxide synthase (iNOS), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β), as well as M2 type macrophage markers arginase1 (Arg1) and IL-10 were detected using real-time PCR andWestern blotting. High fat diet feeding plus multiple intraperitoneal injections of low dose streptozotocin (STZ) were used to induce type II diabetes rat model. After blood glucose level has been stable for five weeks, two fullthickness skin wounds with the diameter of 1cm were made on the back of DM rats. Wounds were randomly assigned to being treated with insulin (0.2 U insulin /20 μL saline) or saline (20 μL saline) using the random number table. Characteristics of macrophagephenotypes were observed 3, 7, and 25days after wounds were made. Normal rats (n=3) served as controls. Results · After being cultured with high glucose, the mRNA levels of M1 markers iNOS and TNF-α were up-regulated in LPS-induced THP-1 cells, while the mRNA levels of M2 markers Arg1 and IL-10 were down-regulated.Afterbeing treated with insulin for 6 h, mRNA levels of iNOS and TNF-α weredown-regulated, protein levels of iNOS, IL-1β were down-regulated too, while mRNAand protein levels of Arg1 and IL-10 were up-regulated. In addition, the expression level of phosphorylated NF-κB-p65 was significantly increased after high glucose culture and was significantly decreased after insulin intervention. Compared to normal rat skin wounds, the expression of iNOS in macrophages was significantly increased in wounds of diabetic rats. The expression of iNOS in macrophages was high in saline treated wounds 3 and 7 days after the wounds were made and the expression of Arg1 was low 25 days after the wounds were made. In insulin treated wounds, the expression of iNOS started to decrease on day 7 after the wounds were made and the expression of Arg1 was significantly higher than that in saline treated wounds on day 25 after the wounds were made. Conclusion · Insulin can induce macrophage phenotype transformation from M1 to M2 under high glucose condition and the mechanism may be associated with the phosphorylation of NF-κB-p65.

AIM: To evaluate the protective effects of mycophenolate mofetil on the kidneys of type 2 diabetic rats and discover their mechanisms. METHODS: Wistar rats were divided into three groups, such as normal control rats, diabetic rats, and diabetic rats in the treatment with mycophenolate mofetil (MMF, 15 mg?kg -1 ?d -1 ). Thirteen weeks later, urinary albumin excretory rate (UAE), creatine clearance (Ccr), blood glucose, blood insulin and blood lipid were measured, and kidney pathology was observed. Inmmunohistochemistry was used to analyze the expression of CTGF, ColI and ColⅢ. RESULTS: Mycophenolate mofetil decreased UAE, Ccr and reduced glomerular volume. The expression of CTGF and deposition of ECM decreased after diabetic rats received mycophenolate mofetil. CONCLUSION: Mycophenolate mofetil can protect the kidney of diabetic rats. Its mechanism may be related to the decrease of CTGF expression and extracellular matrix deposition in renal tissue.

Objective To observe the effect of intra-articular injection of dehydroepiandrosterone (DHEA)on the experimental osteoarthritis in rabbits and study the mechanism.Methods Forty rabbits un derwent unilateral anterior cruciate ligament transection(ACLT)and then divided into two groups randomly. 100?mol/L DHEA resolved in the dimethylsulphoxide were injected into the knees of experimental rabbits 4 weeks after transection,once a week for five weeks.Rabbits in the control group were treated under the same schedule using dimethylsulphoxide.All rabbits were killed 9 weeks after ACLT and the knee joints were evalu- ated by gross morphology and histology.The mRNA expression of metalloproteinases-3(MMP-3),tissue in- hibitor of metalloproteinases-1(TIMP-1)and interleukin-lbeta(IL-1?)in the cartilage and synovium was analyzed using reverse transcription polymerase chain reaction(RT-PCR).Results Gross morphologic in- spection and histological evaluation showed that the extent and grade of cartilage and synovium damage in the experimental group were less severe than the control group.The mRNA expression of MMP-3 in cartilage and synovium decreased significantly in the experimental group(both P＜0.05).The mRNA expression of TIMP-1 in cartilage and synovium increased significantly in the experimental group compared with that in the control group(both P＜0.05).No significant difference of IL-1?mRNA expression in cartilage was found between the experimental and the control groups(P＞0.05).The mRNA expression of IL-1?in the synovium was signifi- cantly suppressed in the experimental group compared with that in the control group(P＜0.01).Conclusion DHEA protects against cartilage degradation,alleviates synovium inflammation and inhibits the progression of osteoarthritis in the experimental model.Down-regulation of MMP-3 and up-regulation of TIMP-1 in cartilage and synovium and IL-1?in the synovium may be the mechanism of the protective effect of DHEA on os- teoarthritis.

Objective:To explore the regulatory role of exosomes in calcification of rat vascular smooth muscle cells (VSMC) induced by high phosphorus.Methods:VSMC (A7r5 cells) were cultured in vitro and randomly divided into three groups: normal phosphorus group (0.9 mmol/L), high phosphorus group (2.6 mmol/L) and high phosphorus exosomes induction group (i.e. the exosomes extracted from high phosphorus group were added to VSMC in normal culture). Until the 7th day of culture, the culture medium of normal phosphorus group and high phosphorus group obtained during the change of cell culture medium was collected, and the precipitate was obtained by ultracentrifugation and suspended by phosphate buffered saline. The protein content of the precipitate was determined by BCA protein quantitative method. The precipitates were identified. The structure and size of exosomes were observed by transmission electron microscope. The exosomes marker proteins tumor susceptibility gene 101 protein (TSG101) and CD9 were detected by Western blotting. The miRNA in exosomes was extracted, and the expression of related miRNA (miR-30b, miR-204, miR-211) were observed by real-time quantitative PCR. After 7 days of cultivation, the exosomes uptake process of VSMC in high phosphorus exosomes induction group was observed. The calcium deposition was detected by Alizarin stain, and the calcium content was detected by O-cresol complex copper. The content of alkaline phosphatase was detected by colorimetry. The protein expression of Runx2 was quantified by Western blotting. Results:(1) The precipitate obtained by ultracentrifugation of the cell culture fluid was identified as exosomes by electron microscopy morphology. Western blotting confirmed that the expression of the exosomal marker proteins TSG101 and CD9 were positive. (2) The exosomes were rich in miRNAs. The expression of miR-30b, miR-204, miR-211, which negatively regulated the transcription of Runx2, was significantly down-regulated in the high-phosphorus group compared with the normal group ( P<0.05). (3) After culturing rat VSMC with high phosphorus for 7 days, calcium salt deposition was obvious. Compared with the normal phosphorus group, calcium content and alkaline phosphatase activity were significantly increased (both P<0.05), and Runx2 expression was also significantly increased ( P<0.05). (4) Added the obtained high-phosphorus exosomes to the normal cultured VSMC, the exosomes could be taken up by VSMC and successfully induced VSMC calcification. The levels of cell calcification indicators and Runx2 expression were significantly increased. Conclusions:High phosphorus induces calcification of VSMC and promotes the increase of Runx2 expression. The mechanism may be realized by releasing exosomes from VSMC to transmit cell signals.

AIM:Receptor tyrosine kinase (RTK) activation is critical for growth factor-mediated cell proliferation. The present study was designed to determine the effect of the tyrphostin AG1295 and AG1296, a selective blocker of PDGF βand αRTK,on proliferative vitreoretinopathy (PVR) development.METHODS:Rabbit conjunctival fibroblasts (RCF) cells were cultured.The effects of AG1295, AG1296,PDGF-AA and PDGF-BB on RCF proliferation are evaluated by MTT assay.Homologous rabbit conjunctival fibroblasts were injected intravitreally to make animal PVR model, followed by injection of 100μmol/L of AG1295 or AG1296 respectively. The presence of tractional retinal detachment (TRD) was assessed to evaluate the effect of AG1295 and AG1296 in vivo .Electroretinography and histologic studies were performed after intravitreal injection of AG1295 into untreated eyes to evaluate toxicity. RESULTS: Both AG1295 and AG1296 (10μmol/L) significantly inhibited rabbit conjunctival fibroblast cell growth stimulated by PDGF-AA or -BB in vitro.Development of TRD was significantly reduced (P<0.05) with 100 μmol/L of AG1295 or AG1296 in vivo, but the effect of AG1295 only present till day 14. Inhibitive effect of AG1296 is longer than that of AG1295.No significant histologic or retinal functional damage was found in both drug-treated groups. CONCLUSION: PDGF αand βreceptor specific inhibitor AG1296 and AG1295 attenuated PVR without significant side effects in rabbits, and AG1296 was better than AG1295. The much longer and stronger therapeutic effect from PDGFαreceptor inhibitor indicated that PDGF α receptor is more important in the development of PVR, and inhibition of this pathway could be a useful treatment alternative to prevent PVR.

0.05). The relative bioavailability of tested capsules to reference tablets was (99.3?13.1)% Conclusion Both formulations are of bioequivalence.

To clarify the function and molecular mechanism of miR-155 in myogenic differentiation of C2C12, we constructed adenovirus over-expression vector of miR-155, then C2C12 cells were infected by adenovirus and induced myogenic differentiation. First, we observed the morphology of C2C12 after differentiation. Then the mRNA and protein expressions of myogenic markers (MyoD, MyoG and MyHC) were detected by qPCR and western blotting. Subsequently, the dual luciferase reporter gene assay was carried out to validate putative target gene (TCF4) of miR-155. Meanwhile, mRNA level of TCF4 was analyzed after over-expressing miR-155. The results show that over-expressed miR-155 reduced myotubes formation. Moreover, the mRNA and protein expression of MyoG and MyHC decreased significantly (P < 0.01). Further research demonstrated miR-155 bound the one (4532-4538) of three putative sites (1487-1493,1516-1522, 4532-4583) of TCF4 by luciferase reporter gene assay and the mRNA level of TCF4 decreased notably (P < 0.05). The data suggest that miR-155 inhibited myogenic differentiation of C2C12 through targeted TCF4.
Animals ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; Cell Differentiation ; Cell Line ; Genetic Vectors ; Mice ; MicroRNAs ; genetics ; Myoblasts ; cytology ; Myogenin ; genetics ; metabolism ; Myosin Heavy Chains ; genetics ; metabolism ; RNA, Messenger ; genetics ; Transcription Factor 4

OBJECTIVEThis study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.

METHODThe HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.

RESULTThe results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.

CONCLUSIONThis method is able to be a scientific basis of quality assessment according to its convenient and reliable.