AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.

OBJECTIVE: To discuss the effect of Sanbi granules on type Ⅱ collagen induced arthritis (CIA) rats by regulating the TLR4/MAPKs/NF-κB signal pathway. METHODS: Sixty of Wistar rats were randomly divided into normal control group (CTL group, n=10), model group (n=10), positive control group (n=10) and low dose of Sanbi group (n=10), middle dose of Sanbi group (n=10), high dose of Sanbi group (n=10). The collagen induced arthritis (CIA) model of rats was adopted and treated for 20 days by intragastric administration from 2 weeks after primary immune. After exposure to sanbi for 35 d, the rats status, paw swelling, arthritis index (AI) and pathological change of synovial tissue were observed. The serum IL-1β, IL-6, tumor necrosis factor α (TNF-α) levels were detected by ELISA. And the expressions of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB) (p65), p-NF-κB (p65), p38, p-p38, extracellular signal-regulated kinase 1/2 (ERK1/2), p-ERK1/2, c-Jun NH2-terminal kinase (JNK), p-JNK mRNA or proteins in synovial tissues were detected by qRT-PCR and Western blot. RESULTS: At the end of experiment, compared with model group, the paw swelling degree and arthritis index (AI) of CIA rats in DXM group and low, middle, high dose of Sanbi groups were lower (P<0.05). Synovial tissue inflammatory of rats in high dose of Sanbi group changed obviously by H&E staining. The serum IL-1β, IL-6, TNF-α of CIA rats in DXM group and low, middle, high dose of Sanbi groups were lower than those in model group (P<0.05). And there was no difference of the serum IL-1β, IL-6, TNF-α between DXM group and high dose of Sanbi group (P>0.05). Besides, compared with CTL group, TLR4, p-NF-κB (p65), p-p38, p-ERK1/2, p-JNK mRNA and proteins in synovial tissues of CIA rats in model group, DXM group and low, middle, high dose of Sanbi groups were higher (P<0.05). And these mRNAs and proteins in DXM group and low, middle, high dose of Sanbi groups were lower than these in model group, particularly in DXM group and high dose of Sanbi group (P<0.05). CONCLUSION: There are significant evidences that Sanbi granules could protect joint synovial tissues injury by down-regulation TLR4/MAPKs/NF-κB signal pathway on CIA rats.

Ulcerative colitis （UC） is one of the chronic refractory inflammatory bowel diseases characterized by abdominal pain， diarrhea， and mucus， pus and blood in the stool. In recent years， with changes in human life style and improvements of the diagnosis， the incidence and prevalence of UC have been increasing. The pathogenesis of UC is closely related to intestinal mucosal immune dysfunction， intestinal flora disturbance， and abnormal bile acid secretion. Patients with UC have abnormal bile acid secretion and intestinal flora imbalance. A large number of studies have found that abnormal bile acid secretion inhibits immune function， affects signal transduction， and destroys the intestinal mucosal barrier. Intestinal flora disturbance has an important impact on the occurrence and development of inflammation， immune homeostasis， and stress. Bile acids indirectly or directly affect the structure and function of intestinal flora， and at the same time， they produce secondary bile acids under the modification of intestinal flora， entering the liver through enterohepatic circulation. Therefore， the complex dialogue mechanism of bile acid-intestinal flora axis is closely related to the occurrence and development of UC. Based on the basic theory of traditional Chinese medicine（TCM） and clinical research， it is found that emotion is an important factor that induces this disease， spleen and stomach weakness is the root of the disease， and liver depression and spleen deficiency are the key pathogenesis of UC. Combined with modern medicine and molecular biology research， it is believed that abnormal secretion of bile acids is a microscopic manifestation of liver depression in TCM， and intestinal flora disturbance is the biological basis of spleen deficiency. In the pathogenesis of UC， the imbalanced bile acid-intestinal flora axis is consistent with the pathogenesis of liver depression and spleen in TCM. The exploration of the biological connotation of the pathogenesis of UC with liver depression and spleen deficiency from the perspective of bile acid-intestinal flora axis can better explain the scientific nature of its pathogenesis， which provides new clinical solutions and reliable references for studying the pathogenesis of UC with liver depression and spleen deficiency and finding representative prescriptions to prevent and treat this disease.