Molecular docking technology and network pharmacology based on Rhapontici Radix-Cremastrae Pseudobulbus drug pair in treating breast cancer.

OBJECTIVE: Network pharmacology is a bioinformatics-based research strategy for identifying the mechanisms of drugs and promoting drug development. This study used network pharmacology to investigate the mechanism of the Loulu-Cremastrae Pseudobulbus drug pair treating breast cancer (BC). MATERIALS AND METHODS: The ingredients and potential targets of the drug pair were searched with Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCSMP). National Center for Biotechnology Information (NCBI) and gene cards were used to search the targets of BC. Networks of "drugs-components-targets" and protein-protein interaction were constructed through Cytoscape. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were carried out through common targets. Using AutoDock tool, molecular docking was performed to verify the binding between key targets and compounds. RESULTS: Finally, we selected 6 active compounds from the drug pair. A total of 61 targets were associated with the drug pair, and 15,295 targets were related to BC. 55 common targets were obtained after the intersection. The key targets included Transcription factor Jun (JUN), Heat shock protein HSP 90-alpha (HSP90AA1), and Caspase-3(CASP3). 327 terms were obtained by GO analysis. 78 pathways (p < 0.05) were identified through KEGG analysis. Molecular docking indicated that important compounds combined well with key targets. CONCLUSIONS: Various active compounds, including beta-sitosterol, 2-methoxy-9,10-dihydrophenanthrene-4,5-diol, and stigmasterol, can regulate multiple signaling pathways related to BC, such as the estrogen and prolactin signaling pathways, playing therapeutic roles in BC.

[Clinical study of transjugular intrahepatic portosystemic shunt with Viatorr stent in 43 cases].

Objective: To investigate the clinical efficacy and safety of transjugular intrahepatic portosystemic shunt (TIPS) with the use of Viator stent in the treatment of cirrhotic portal hypertension. Methods: 43 cases with cirrhotic portal hypertension were implanted with Viatorr stent during TIPS procedure from March 2016 to August 2018. Serological indicators, color Doppler ultrasound, gastroscopy, rebleeding, ascites and hepatic encephalopathy were regularly followed up. Portal venous pressure, liver and kidney function, coagulation indexes were compared by t-test. Stent patency rate, hepatic encephalopathy incidence, rebleeding rate and survival rate were calculated by Kaplan-Meier curve. Results: TIPS procedure success rate was 100% in all patients. Portal pressure gradient was decreased from (25.57 ± 5.50) mmHg to (9.76 ± 2.92) mmHg before and after operation. Alanine aminotransferase (ALT) was significantly higher at 1 month after operation than before operation, but there was no significant difference between 3 and 6 months after and before operation. Total bilirubin, serum ammonia and prothrombin time at 1, 3, and 6 months after operation were higher than before operation. Albumin had no significant change compared with before operation, and creatinine and urea nitrogen were lower than before operation. The cumulative rebleeding rates at 12 and 24 months after operation was 0% and 9%, respectively. Of the 26 patients with ascites, 22 cases (84.6%) had complete disappearance of ascites and 3 (11.5%) had significant decrease of ascites. The cumulative incidence of hepatic encephalopathy at 3, 6, 12 and 24 months after surgery was 11.6%, 17.3%, 21.9% and 21.9%, respectively. The cumulative incidence of stent dysfunction at 12 and 24 months after surgery was 5.6% and 23.7%, respectively. The cumulative survival rate at 12 months and 24 months after surgery was 91.9%. Conclusion: TIPS procedure with Viatorr stent can effectively reduce portal pressure and rebleeding rate, improve intrahepatic shunt patency rate, and will not increase the risk of postoperative hepatic encephalopathy, and has a higher cumulative survival rate.

Preparation and immune activity analysis of H5N1 subtype avian influenza virus recombinant protein-based vaccine.

Avian influenza is a severe disease among farmed poultry and free-living birds and a constant threat to the commercial chicken industry around the world. Hemagglutinin (HA) is the major immunogen on the envelope of influenza A virus and is the predominant inducer of neutralizing antibody. To obtain the bioactive antigen proteins in large quantities, a new protein expression vector pBCX was constructed, which is based on the pET32a vector. The HA gene of the H5N1 subtype of avian influenza virus (AIV) was inserted into the pBCX vector and expressed efficiently in Escherichia coli BL21 (DE3). Fused expression of the exogenous gene and msyB produced a 97-kDa msyB-HA fusion protein. Sodium dodecyl sulfate-PAGE combined with scanning analysis demonstrated that the msyB-HA fusion protein accounted for 29.5% of the total bacterial protein, 90.5% being soluble. The msyB-HA fusion protein was purified with nondenaturing 50% Ni-NTA column chromatography, and the result showed that 24 mg of purified msyB-HA fusion protein could be obtained from 1 L of induced expression bacterial culture medium. The comparative results in the present study showed that pBCX was superior to pET32a as a protein expression vector. Western blotting showed the recombinant msyB-HA (rHA) to have better antigenic activity, which may be the result from the better posttranslation protein modification and folding in the pBCX expression system. With the rHA fusion protein as antigen, we successfully prepared and screened specific monoclonal antibodys against the H5N1 subtype AIV, which indicated that the rHA had antigen epitopes and biofunctions. The immune test confirmed that the rHA protein vaccine could also induce high neutralizing antibodies, and the AIV challenge test proved that the rHA protein-based vaccine could prevent the corresponding infection. This study demonstrates that the recombinant HA protein produced by the pBCX expression system could be used as a recombinant protein-based vaccine and has potential for further development for diagnosis.