BACKGROUND:Studies have found that Wudi Dan can suppress local inflammatory response of the lesioned joints to protect the articular cartilage. OBJECTIVE:To verify the effect of Wudi Dan on chondrocyte viability and apoptosis as wel as the therapeutic effect on osteoarthritis. METHODS:Rat chondrocytes were cultured in serum medium containing Wudi Dan, and the effects of Wudi Dan on cel viability and apoptosis were observed by comparison with the control group. Rabbit model of knee osteoarthritis was constructed using modified Hulth method. Rabbit models were divided into two groups:Wudi Dan group treated with Wudi Dan and control group treated with normal saline, twice a day, consecutively for 4 weeks. Therapeutic effect of Wudi Dan on knee osteoarthritis was observed;cel viability and apoptosis were observed under microscope;the levels of interleukin-1 and matrix metal oproteinase-3 were determined using immunohistochemical method. RESULTS AND CONCLUSION:The apoptotic rate of chondrocytes was significantly lower in the Wudi Dan group than the control group. Pathological findings of the rabbit knee joints showed that the control group had more severe damage to the articular cartilage than the Wudi Dan group. Immunohistochemical staining revealed that in the Wudi Dan group, the cytoplasm and extracel ular matrix were colored light and there were a smal number of positive cel s as wel as low expression of interleukin-1 and matrix metal oproteinase-3. The results suggest that Wudi Dan can effectively protect against articular cartilage lesions, reduce inflammation, and have a good therapeutic effect on osteoarthritis. Its mechanism may be related to inhibition of chondrocyte apoptosis, reduction of cytokine production and inhibition of protein expression of matrix metal oproteinase.

To investigate the role of Toll like receptor (TLR) in the activation of dendritic cells (DC) during early Plasmodium yoelii infection of the lethal strain 17XL (P.y 17XL), susceptible BALB/c and resistant DBA/2 mice were infected by i.p.injection of the P.y l7XL-parasitized erythrocytes, and the parasitemia of individua1 mice was monitored by the microscopic examination of blood smear stained with Giemsa.Mice from norma1 and infected groups were sacrificed on 0,3 and 5 days post-infection to collect their spleen cells.And the expressions of TLR-9 and TLR-4 on the cell surface of DCs in spenonocytes of these two strains of mice were assayed by applying flow cytometry to quantitatively analyze the percentages of CD11c+TLR9+ DCs and CD11c+TLR4+ DCs. It was found that the population of CD11c+DCs expressing TLR9 was significantly increased on day 3 and peaked on 5 p.i. in BALB/c (P<0.01) and DBA/2 mice(P<0.01). However, there was no statistical significance between these two strains of mice. Meanwhile, there was no change on the population of CD11c+ DCs expressing TLR4 in BALB/c and DBA/2 mice. These results indicate that TLR9 may contribute to the DC activation during early stages of P.y17XL infection.

Objective To investigate the effect of MRE11 on the proliferation and apoptosis of esophageal squamous cancer cells and its molecular mechanism. Methods MRE11 expression was downregulated by MRE11 siRNA transfection in esophageal squamous cancer cells. The AKT agonist SC79 (0, 0.1, 0.5, 1, 1.5, 1.8, 2 μg/ml) were used to treat cells with MRE11 inhibition for 24 h. Overexpression vector pcDNA.3.1-c-myc was constructed and co-transfected cells with MRE11 siRNA. Western blot method was used to detect the protein expressions of MRE11, p-AKT and c-myc in esophageal squamous cancer cells Ec9706 and TE-1. The Annexin-V FITC/PI kit was used to detect the apoptosis of Ec9706 and TE-1 cells; the activity of caspase-3 was detected by the Caspase-3 activity detection kit; the proliferation of Ec9706 and TE-1 cells was tested by the BrdU method. Results The protein expressions of MRE11 in Ec9706 and TE-1 cells were significantly increased, compared with human esophageal epithelial Het-1A cells. After MRE11 siRNA transfection, AKT phosphorylation and the protein expressions of MRE11 and c-myc were significantly decreased in esophageal squamous cancer cells. MRE11 inhibition significantly promoted the apoptosis and caspase-3 activity in Ec9706 and TE-1 cells, while inhibited the proliferation of Ec9706 and TE-1 cells. SC79 (1.5, 1.8 and 2 μg/ml) significantly increased AKT phosphorylation in MRE11-suppressed esophageal squamous cancer cells, and reversed the inhibitory effects of MRE11 inhibition on c-myc protein expression and cell proliferation and the promoting effect on cell apoptosis. Overexpression of c-myc inhibited the inhibitory effect of MRE11 down-regulation on cell proliferation and the promotion on caspase-3 activity. Conclusion MRE11 inhibition could effectively inhibit the proliferation of esophageal squamous cancer cells and promote cell apoptosis by regulating AKT and c-myc.

Objective To study the clinical,pathological and molecular biology features of myotonic myopathies.Methods Eighty-one patients with myotonic myopathies,admitted to our hospital from June 2005 to June 2018,were chosen in our study.All patients accepted clinical and skeletal muscle pathology examination,and genetic features of 55 patients were analyzed by molecular biological method.Results (1) All patients suffered from typical myotonia,and electromyography shows typical myotonic discharges;47 patients exhibited myotonic dystrophy (DM) and 34 patients exhibited non-myotonic dystrophy (NDM).(2) In muscle biopsy of DM,typical central nuclei,pyknotic clumps and sarcoplasmic masses were observed;and characteristic pathological changes were not observed in muscle biopsy of NDM.(3) Totally,32 DM1 patients,3 DM2 patients,9 MC patients and 5 paramyotonia congenita patients were confirmed by molecular biology technology;7 independent mutations in the CLCN1 gene and 3 independent mutations in the SCN4A gene were novel mutations.Conclusions (1) Myotonic myopathies are some single gene inheritance diseases with multisystem disorders and their main symptoms include myotonia.(2) Skeletal muscle biopsy is a trustworthy method for definite diagnosis of myotonic myopathies;gene analysis is the gold standard for diagnosis and classification ofmyotonic myopathies.

Objective To explore the anti-tumor mechanism of saffron (Crocus sativus L.) by network pharmacology and reverse molecular docking techniques. Methods The main chemical components of saffron were obtained by searching published literature and TCMSP database. The potential targets of these components were predicted using PharmMapper server. The corresponding target genes were identified from UniProt database. The underlying anti-tumor targets of saffron were obtained by mapping the disease genes of cancer or tumor with GeneCards, OMIM and TTD databases. Cytoscape software was used to construct the action target network of saffron active components. The protein-protein interaction analysis was performed by String database, and the GO function and KEGG pathway enrichment analysis were performed by Metascape platform. Finally, molecular docking was performed to evaluate the binding of main components with their potential targets. Results A total of 9 active ingredients in saffron including quercetin, kaempferol, isorhamnetin, picrocrocin and crocin I, were identified, which might act on 37 key targets including AKT1, CCND1, MMP9, EGFR, TP53, involved in P53, TNF and other signaling pathways. Molecular docking indicated modest binding potency through hydrogen bonding, and hydrophobic interactions. Conclusion The anti-tumor effect of saffron was evaluated via the network of components-targets-pathways, which might provide a foundation for further research.