Aim To establish and evaluate the simple and effective animal model of rheumatoid arthritis in SD rats induced by heat-killed mycobacterium tuberculosis H37Ra(Mtb).Methods SD rats were immunized s.c.at the base of the tail with Mtb(1 mg/rat) in mineral oil,and then body weight,blood routine,clinical signs of arthritis were observed regularly.And also hind paw volume in SD rats with AA was tested.Meanwhile,T-lymphocyte subset was determined by flow cytometry in peripheral blood,the level of TNF-?in serum and IL-1 and IL-6 and vascular endothe-lial growth factor(VEGF) in joint tissue were tested by ELISA.The ankle pathological change and radiographic evidence of the hind paws of SD rats were observed to evaluate the AA model effects and availability in SD rats.Results When compared with normal control rats,the total number of white blood cells,platelets and monocytes in peripheral blood and ratio of CD4/CD8 was markedly raised after immunization with Mtb.But body weight of model rats was significantly low from day 14 to 28 after injection of Mtb.The inflammatory arthritic lesions such as edema and erythema in the paws appeared after 9～10 d.The peak of inflammation was 14～16 d.The level of TNF-?in serum and IL-1,IL-6 and VEGF in joint tissue was significantly higher in SD rats with Mtb than that in normal control rats at d 28.The histopathological examination revealed cellular infiltration,synovial and cartilage hyperplasia in ankle joints of SD rats.Conclusions The model induced by Mtb in SD rats is more similar in clinical characterization of rheumatoid arthritis in human being.It is a more efficient and simpler manipulative procedure for screening of new anti-arthritis drugs and experimental studying of anti-rheumatoid arthritis.

In order to search for a more potent and less toxic antifungal agent, five novel sulconazole analogues were synthesized by changing 1-imidazolyl and p-chlorobenzyl in sulconazole structure. The results of preliminary biological tests show that analogues Ⅰ and Ⅱ possess more potent antifungal activities and wider antifungal spectra than sulconazole.

Gastroparesis is a gastric disease characterized by delayed gastric emptying."Vomiting"is the main clinical manifestation of this disease,while"atrophy"indicates dysfunction of the stomach.Based on the theory of"atrophy,dyspnea and vomiting are ascribed to the upper part",originating from the Chapter of"Zhi Zhen Yao Da Lun"from the Suwen(Basic Questions)section this paper explored the pathogenesis and treatment idea of gastroparesis from the function of the lung.It is put forward that failure of lung qi in dispersion,so the lung qi cannot maintain the circulation of harmonizing qi and blood,leading to malnutrition of stomach and loss of stomach receiving food and drink function.Melancholy impairing lung,so lung cannot govern diffusion,leading to stomach governing the disfunction of dredging and descending.Impaired depurative descending of lung qi,so the large intestine and stomach are lost in the alternating operation of deficiency and excess,leading to stomach governing the disfunction of transportation and transformation.Therefore,the treatment approach is proposed to improve the stomach's function to receive food and drink,transport and transform and promote the recovery of gastric motility by replenishing and restoring lung qi,regulating emotions and catharsis and smoothing lung.Analyzing the effect of function of the lung on gastric motility is of great value for expanding the application of traditional Chinese medicine in gastrointestinal motility disorders,and digging new connotations of classic theories in modern clinical practice.

Objective To clone the HnGUA gene from Hirudo nipponia and conduct bioinformatics analysis,protein prokaryotic expression analysis and gene differential expression analysis.Methods Based on the transcriptome data of H.nipponia in the previous study,the full-length cDNA of HnGUA was cloned by rapid amplification of cDNA ends(RACE),and bioinformatics analysis was performed.The prokaryotic expression vector was constructed,transformed into Escherichia coli BL21(DE3)competent cells and the expression of recombinant protein was induced by IPTG.The qPCR was used to further analyze the tissue-specific expression of HnGUA.Results The size of HnGUA gene was 504 bp,containing an open reading frame(ORF)of 231 bp and encoding 76 amino acids.Its protein molecular weight and isoelectric point are 8.17 kDa and 4.44,respectively.Multiple sequence alignment analysis showed that HnGUA was highly homologous to genes in other leech species that encode inhibitory proteins.The results of the prokaryotic expression analysis showed that the constructed pET32a-HnGUA vector could be successfully expressed in E.coli BL21(DE3),and the SDS-PAGE results showed that the induced recombinantly expressed HnGUA protein was around 6 kDa,which was basically consistent with the predicted protein size.The results of the Real-time PCR revealed spatial and temporal differences in the expression profiles of HnGUA,with high levels of expression detected in the skin and crop tissues.Conclusion This study represents the first successful cloning of the HnGUA gene from H.nipponia and the expression of the corresponding recombinant protein in E.coli.It provides a foundation for future exploration of the biosynthetic pathways and molecular regulatory mechanisms of active small anticoagulant molecules in leeches.