The antimicrobial activity of nitropropane compound 791224 against several phytopachogenic fungi and Fusarium was studied in vitro. Its inhibitory effect on mycelial growth of Fusarium oxysporum f. sp. vasinfectum (Atk.) Snyder et Hansen and Fusarium graminearum Schwabe showed a much stronger than that on the other phytopathogenic fungi tested. The inhibition of 791224 on all tested Fusarium showed excellent. Its inhibitory effect values ranged from 54.2 to 100% at 5 ppm for 12 tested Fusarium whereas the respective values of carbendazim ranged from 0 to 27.3%. The inhibitory effect of 791224 at 5 ppm on Fusarium lateritium Nees reached to 100% whereas for carbendazim it was 0, i.e.100 times lower than that of 791224. In green house, the control efficiency of 791224 on Fusarium solani sothora japonico H. has been tested and showed a promising result.

Objective To analyze the changes of the intima-media thickness(IMT)of carotid and femoral arteries, serum advanced glycosylation end-products(AGEs),and AGEs soluble receptor(sRAGE)after intensively controlling blood glucose, blood pressure, and lipid. Methods One hundred and thirty-two type 2 diabetic patients were divided into 3 groups and followed for 5 years: 20 patients were treated with intensive control of blood glucose and blood pressure, 80 patients with intensive control of blood glucose, blood pressure, and lipid; and 32 patients with conventional therapy. AGEs, sRAGE, and IMT of carotid and femoral arteries were measured and compared among different groups. Results The IMT of carotid and femoral arteries and serum level of AGEs were significantly decreased after intensive treatment. The ratio of sRAGE and HbA1C(sRAGE/HbA1C)were negatively correlated with the mean of HbA1Cin the past five years(r=-0.417, P＜0.001)and the fluctuation of HbA1C(r=-0.309,P＜0.001). Multinomial regression analysis showed that AGEs were the important risk factors of IMT of femoral artery(β=0.152,P=0.068). Conclusion Intensive treatment is significant in controlling the growing IMT of carotid and femoral arteries, while decreasing serum level of AGEs.

OBJECTIVE: To explore the effect of decoction (DGNTD) on cell apoptosis and TNF receptor super family 6 (Fas)/caspase-8 pathway in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). METHODS: FLS isolated from the synovial tissue of RA patients were cultured and identified using immunofluorescence staining. The cells were treated with 10% blank serum (blank control group), 10% sera containing low, moderate or high doses of DGNTD, or 20 μmol/mL KR-33493 (a Fas inhibitor) combined with 10% serum containing high-dose DGNTD. MTT assay was used to detect the proliferation of the cells after the treatments. Apoptosis of the cells was detected at 48 h in each group using Hoechst 33342 staining and flow cytometry with annexin V-FITC/PI staining. The mRNA and protein expressions of Fas, FADD, caspase-8 and caspase-3 in the cells at 48 h were detected using qPCR and Western blotting. RESULTS: Immunofluorescence staining identified the cultured cells as FLS. Treatment with DGNTD-containing sera significantly inhibited the proliferation of FLS, and the inhibitory effects were enhanced as the dose and intervention time increased ( < 0.05). Hoechst 33342 staining and flow cytometry showed that the sera containing different doses of DGNTD significantly promoted apoptosis of FLS ( < 0.05). The expression levels of Fas, FADD, caspase-8, and caspase-3 at both mRNA and protein levels were significantly increased in the cells after treatment with different doses of DGNTD-containing sera ( < 0.05). The application of KR-33493 obviously reversed the effects of DGNTD on the FLS ( < 0.05). CONCLUSIONS DGNTD can induce apoptosis of the FLS by activating Fas/caspase-8 signaling pathway.
Apoptosis ; Arthritis, Rheumatoid ; Caspase 8 ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; Synovial Membrane ; Synoviocytes

ObjectiveTo evaluate the animal model of rheumatoid arthritis （RA） induced by inactivated Mycobacterium tuberculosis （Mtb） from inflammation， apoptosis and autophagy， compare the rat RA model of wind-damp-heat arthralgia （FSR） induced by Mtb with the rat RA model of adjuvant arthritis （AA）， and provide experimental evidence for improving the disease-syndrome combined model and developing drugs for the prevention and treatment of RA. MethodThirty 6-week-old SD rats were randomly assigned into 3 groups： a normal group， an AA group and a FSR group， with 10 rats in each group. The RA rat model was established by injection of Mtb suspension （1 mg/rat）， and the rats in the normal group were injected with the same volume of normal saline. The invention in the FSR group lasted for 16 days. The general conditions， body weight， spleen index， swelling of ankle joints， arthritis index （AI）， and the symptoms of arthritis in the hind feet of the rats in each group were observed and measured. The expression levels of serum interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of IL-1β， TNF-α， apoptosis-related protein B cell lymphoma-2 （Bcl-2） and Bcl-2 associated X protein （Bax）， autophagy-related protein 1 light chain 3 （LC3）， autophagy key molecule yeast Atg6 （Beclin1），and p62 mRNA expression levels were detected by Real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of Bcl-2， Bax， LC3， Beclin1 and p62 were detected by Western blot. ResultCompared with those of the normal group， the rats of AA and FSR groups grew slowly and presented dull hair， soft or loose stool， slow movement， swelling of spleen， redness and swelling of ankle joints， increased AI， and histopathologic changes in the synovium and ankle joints. Moreover， the modeling elevated the levels of IL-1β and TNF-α in the serum， up-regulated the mRNA levels of IL-1β and TNF-α in the synovium， up-regulated the mRNA and protein levels of Bcl2， LC3， Beclin1， and down-regulated the mRNA and protein levels of Bax and p62. Compared with the AA group， the FSR group showed severe symptoms， slowly increased body weight （P<0.01）， early appearance of obvious redness and swelling of ankle joints， increased AI （P<0.05）， increased spleen index （P<0.05）， and obvious pathologic changes of synovial tissue and ankle joints， the inflammatory infiltration in the synovial tissue， and the damage of joint structure. Moreover， the FSR group had higher expression levels of IL-1β， TNF-α， Bcl-2， LC3， and Beclin1 （P<0.05） and lower expression levels of Bax and p62 than the AA group （P<0.05）. ConclusionThe animal model of RA （syndrome of wind-damp-heat arthralgia） can be induced successfully with the symptoms consistent with clinical manifestations of RA. The FSR group has lower apoptosis level and higher autophagy level than the AA group， which indicates that the FSR group is more appropriate for the research on the RA with the syndrome of wind-damp-heat arthralgia.

ObjectiveTo explore the mechanism of Danggui Niantongtang （DGNT） against adjuvant arthritis （AA） rats with wind-dampness-heat arthralgia by quantitative proteomics. MethodSixty SD rats were randomly divided into normal group， model group， angelica came pain soup low， medium and high dose group and methotrexate （MTX） group， each group of 10， only the rat tail root subcutaneously inactivated mycobacterium tuberculosis （Mtb） of adjuvant to build model of AA， artificial climate box intervention 16 d rheumatic fever bi syndrome model is set up， building the day began to drug intervention， The intervention lasted for 28 days. The proteins of synovial tissues in experimental rats were extracted. The differential proteins in the medium-dose DGNT group and the model group were detected and analyzed by 4D label-free quantification （4D-LFQ） proteomics. The differentially expressed proteins associated with mitochondrial pathway apoptosis were verified by immunohistochemistry and Western blot. ResultA total of 4 756 proteins were identified from rat synovial tissues， of which 4 234 proteins contained quantitative information. There were 814 differential proteins between the model group and the DGNT group. As revealed by Gene Ontology （GO） and Kyoto Encyclopedia of Gene and Genome （KEGG） enrichment analyses， DGNT had an effect on the synovial proteome of AA rats with wind-dampness-heat arthralgia， and the differential proteins were enriched in the regulation of the immune system， response to acute inflammation， and apoptosis regulation. As demonstrated by the results of immunohistochemistry and Western blot， compared with the model group， the DGNT groups and the MTX group showed increased protein expression of B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax） and cytochrome C （Cyt C）（P<0.05， P<0.01）， reduced Bcl-2 level （P<0.05， P<0.01）， elevated level of cleaved cysteinyl aspartate-specific protease 9 （Caspase-9）/Caspase-9 （P<0.01）， and decreased level of phosphorylated protein kinase B （p-Akt）/Akt（P<0.05， P<0.01）. ConclusionDGNT involved multiple targets in the treatment of AA with wind-dampness-heat arthralgia and it may exert its effect in the prevention and treatment by regulating the Akt/Bax/Bcl-2 pathway and promoting the cell apoptosis in the mitochondrial pathway.