Rapid molecular identification of Rana dybowskii by species-specific primers.

Oviductus Ranae is the dried oviduct from Rana dybowskii, a forest frog species with medicinal, tonic, and cosmetic properties. Due to the high price and resource shortage, counterfeit varieties of Oviductus Ranae often appear in the market. However, traditional identification methods cannot accurately differentiate between Oviductus Ranae and its adulterants. In this study, a rapid molecular identification method has been established. The method involves extracting genomic DNA in just 30 s using filter paper purification, species-specific rapid polymerase chain reaction (PCR) amplification, and finally, fluorescence detection of the products. It can accurately identify Oviductus Ranae and its three common adulterants in about 30 min, making the process simple, fast, and highly specific.

[Research progress in molecular pharmacognosy of Pogostemon cablin].

Molecular pharmacognosy( MP) is a new interdisciplinary science,which integrates the pharmacognosy and molecular biology,and focuses on the crude drugs' classification and identification,cultivation and protection,and production of active ingredients at the molecular level. Pogostemon cablin is one of the ten major southern medicines in China,MP research on this famous herb has developed on the basis of traditional research methods,and achieved certain results. This article summarized the MP research achievements of P. cablin in recent years,the prospect of this field is also discussed to provide references for the protection,development and utilization of P. cablin resources.

Progress in Application of Nanotechnology in Sorafenib.

Dysregulation of the tyrosine kinase signaling pathway is closely related to tumor development, and tyrosine kinase inhibitors are important targets for potential anticancer strategies. In particular, sorafenib, as a representative drug of multitarget tyrosine kinase inhibitors, has an important clinical status and is widely used for treating various solid tumors and diabetic complications. However, poor aqueous solubility of sorafenib, poor bioavailability of commonly used oral dose forms, poor accumulation at tumor sites, and severe off-target effects that tend to induce intolerable systemic side effects in patients have greatly reduced its therapeutic efficiency and limited its extensive clinical application. To improve the properties of sorafenib, increase the efficiency of clinical treatment, and overcome the increasingly prominent phenomenon of sorafenib resistance, multiple investigations have been conducted. Numerous studies have reported that the properties of nanomaterials, such as small particle size, large specific surface area, high surface activity and high adsorption capacity, make nanotechnology promising for the construction of ideal sorafenib nanodelivery systems to achieve timed and targeted delivery of sorafenib to tumors, prolong the blood circulation time of the drug, improve the utilization efficiency of the drug and reduce systemic toxic side effects. This review summarizes the progress of research applications in nanotechnology related to sorafenib, discusses the current problems, and expresses expectations for the prospect of clinical applications of sorafenib with improved performance.

Sinomenine increases adenosine A2A receptor and inhibits NF-κB to inhibit arthritis in adjuvant-induced-arthritis rats and fibroblast-like synoviocytes through α7nAChR.

Sinomenine (SIN) is a clinical drug for treating rheumatoid arthritis (RA) in China. Our previous study found SIN inhibited inflammation via alpha7 nicotinic acetylcholine receptor (α7nAChR) in macrophages in vitro. Adenosine receptor A2A has anti-inflammatory and immunosuppressive function. However, the mechanisms of SIN acting on α7nAChR and the effect on adenosine A2A receptor (A2A R) in RA are not clear. In the present study, the effects of SIN on adjuvant-induced-arthritis (AIA) rats in vivo and on fibroblast-like synoviocytes (FLSs) in vitro were investigated. Indomethacin (Indo) and methotrexate (MTX), the clinical anti-arthritis drugs, were used as controls. Nicotine (Nic), a specific agonist of α7nAChR, was used as a control for targeting α7nAChR. Alpha-bungarotoxin (α-BTX), the antagonist of α7nAChR or small interference RNA (siRNA) was used to block or knock down α7nAChR. Results showed that SIN decreased arthritis index, hind paw volume, erythrocyte sedimentation (ESR) and serum TNF-α in AIA rats, and α-BTX attenuated the earlier-mentioned effects of SIN and Nic, but not Indo and MTX. The expressions of A2A R in synovium declined in AIA rats, but remarkably increased after the intervention of SIN. The expression of A2A R decreased by LPS or TNF-α, but increased by SIN; cAMP also increased by SIN in FLSs in vitro. SIN inhibited the expression of MCP-1, IL-6, and vascular endothelial growth factor in LPS-induced FLSs. SIN inhibited the activation of NF-κB. Meanwhile, α-BTX or α7nAChR siRNA blocked the earlier-mentioned effects of SIN in FLSs. Results suggested the expressions of A2A R in synovium and FLSs are negatively correlated with the arthritis progression of AIA rats and the activation of FLSs. SIN increases A2A R and inhibits the activation of NF-κB pathway via α7nAChR in AIA rats and FLSs.

[Transformation of antimicrobial peptide fusion gene of cecropin B and rabbit NP-1 to Houttuynia cordata].

OBJECTIVE: To transform the antimicrobial peptide fusion gene of cecropin B and rabbit NP-1(CN) into Houttuynia cordata to improve its antimicrobic capability. METHOD: The fusion gene of CN designed and synthesized artificially was recombined with expression vector pBI121. The recombined vector was transformed to Agrobacterium tumefaciens LBA4404, by which CN gene was transformed to the explants of H. cordata. The transgenic regeneration plantlets were selected by kanamycin and rapid screening PCR. The transgenic plants were identified by PCR-Southern of genomic DNA and RT-PCR. The disease resistances were detected by antibacterial zone trail of leaf extracts to E. coli K12 and infection by Rhizoctonia solani. RESULT: Gene of interesting CN was inserted into genomic DNA and expressed in transformed H, cordata, whose resistance to E. coli K12 and Rh. solani was stronger than that of the non-transformed control. CONCLUSION: The fusion gene CN can improve antimicrobic capability of transformed H. cordata.

Rapid identification of Dendrobium officinale using Loop-Mediated Isothermal Amplification (LAMP) method.

Dendrobium officinale is not only an ornamental plant, but also a valuable medicinal herb that is widely used in traditional Chinese medicine. However, distinguishing D. officinale from other Dendrobium species is usually a difficult task. In this study, we developed a rapid identification protocol for D. officinale using the loop-mediated isothermal amplification (LAMP) method. A set of primers were specifically designed to detect a modified internal transcribed spacer region of D. officinale at 65 °C within 40 min after adding SYBR® Green I, which was used for the detection of D. officinale. Unlike commonly used adulterants, reaction mixtures containing D. officinale DNA changed from orange to green, and this color change was easily observed with the naked eye. Thus, this methodology can be used to accurately differentiate D. officinale from other Dendrobium species, is quick as all D. officinale samples were amplified within 40 min, and specific as samples of the adulterants were not amplified. The specificity of this LAMP-based method was confirmed by testing 17 samples of D. officinale and 32 adulterant samples from other Dendrobium species. This LAMP-based rapid identification method does not require expensive equipment or specialized techniques and can be used in field surveys for accurate and fast on-site identification.