[Anti-inflammatory mechanism of Balanophora involucrata: a network pharmacology and molecular docking-based analysis and verification in lipopolysaccharide-induced RAW264.7 cells].

OBJECTIVE: To investigate the main chemical constituents of Balanophora involucrata and the mechanism of its antiinflammatory effect based on network pharmacology and molecular docking technology. METHODS: Literature reports, Materia Medica, GeneCards and other databases were searched for anti-inflammatory compounds and their targets. String database and Cytoscape 3.7.2 software were used to obtain the protein-protein interaction (PPI) network and the drug-active ingredienttargets network and for GO and KEGG enrichment analyses. Molecular docking was performed using Auto Dock Tools 1.5.6. In an inflammatory RAW264.7 cell model induced by lipopolysaccharide (LPS), the effect of 25, 50, 100, 200 µg/mL Balanophora involucrata extract was tested on the production of inflammatory cytokines and phosphorylation level of PI3K and Akt using ELISA and Western blotting. RESULTS: A total of 318 common targets of drugs and diseases were identified, and the core targets were Src, HSP90AA1 and PIK3CA, involving cancer, PI3K/Akt, MAPK and other signaling pathways as shown by KEGG analysis. Molecular docking showed that both the main active constituents of Balanophora involucrata could spontaneously bind to the core targets. In the inflammatory cell model, treatment with Balanophora involucrata extract significantly inhibited the production of IL-1ß at the concentrations of 100 and 200 µg/mL, reduced IL-6 and TNF-α expressions at the concentrations of 50, 100, and 200 µg/mL, and lowered phosphorylation levels of PI3K and Akt proteins at the concentrations of 25, 50, 100, and 200 µg/mL (all P < 0.05). CONCLUSIONS: The anti-inflammatory mechanism of Balanophora involucrata involves multiple targets and multiple pathways, and its effect is mediated possibly by reducing IL-1ß, IL-6 and TNF-α production and inhibiting phosphorylation levels of PI3K and Akt proteins to suppress the activation of the PI3K/Akt signaling pathway.

Sentinel lymph nodes show profound downregulation of antigen-presenting cells of the paracortex: implications for tumor biology and treatment.

The sentinel lymph node (SN) is the first node on the direct lymphatic drainage pathway from a tumor. Melanoma-associated SNs are the most likely site of early metastases and their immune functions are strikingly down-modulated. We evaluated histologic and cytologic characteristics of 21 SNs and 21 nonsentinel nodes (NSNs) from melanoma patients who had clinically localized (AJCC Stage I--II) primary cutaneous melanoma. SNs showed highly significant reductions in total paracortical area and in the area of the paracortical subsector occupied by dendritic cells. The frequency of paracortical interdigitating dendritic cells (IDCs) was dramatically reduced in SNs, and most IDCs (approximately 99%) lacked the complex dendrites associated with active antigen presentation. The release of immunosuppressive factors from the primary melanoma may induce a localized and specific paralysis in the SN, which prevents the recognition of otherwise immunogenic melanoma antigens by IDCs. This immune paralysis may facilitate the implantation and growth of melanoma cells in the SN. Cytokine therapy may be able to reverse this immune paralysis. These findings have an important practical application in the histopathologic confirmation that a node is truly sentinel. They also offer an hypothesis to explain the failure of the immune surveillance mechanisms to identify and respond to a small primary melanoma that expresses immunogenic tumor antigens.

[Studies on chemical constituents of Ilex Merr].

OBJECTIVE: To further probe into the chemical composition of Ilex hainanensis. METHOD: Compound isolation was affected on the basis of the EtOAc soluble fraction of the ethanolic extract from the leaves of the plant. RESULT: Four compounds were obtained and by physico-chemical and spectroscopic methods identified as ursolic acid, ilexgenin A, ilexsaponin A1 and beta-sitosterol-3-O-beta-D-glucoside. CONCLUSION: All the four compounds were isolated from the plant for the first time.

[Constituents in petroleum ether and ethyl acetate extract fractions of Dracaena cochinensis (Lour.) S.C. Chen].

OBJECTIVE: To study the chemical constituents of Dracaena cochinensis. METHODS: Compounds were separated by solvent extraction, column chromatography, TLC and HPLC, and their structures were determined by spectral analysis. RESULTS: Ten compounds were isolated from the petroleum ether and ethyl acetate fractions of D. cochinensis and identified as n-heptacosane, lophenol, docosanyl ferulate, tetracosanyl ferulate, hexacosanyl ferulate, octacosanyl ferulate, pterostilbene, dioctyl phthalate, butyl isobutyl phthalate and 4'-methoxy-3',7-dihydroxy-flavone. CONCLUSION: All the compounds were isolated from this plant for the first time.

Molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system.

The 2;5 chromosomal translocation is frequently associated with anaplastic large cell lymphomas (ALCLs). The translocation creates a fusion gene consisting of the alk (anaplastic lymphoma kinase) gene and the nucelophosmin (npm) gene: the 3' half of alk derived from chromosome 2 is fused to the 5' portion of npm from chromosome 5. A recent study shows that the product of the npm-alk fusion gene is oncogenic. To help understand how the npm-alk oncogene transform cells, it is important to investigate the normal biological function of the alk gene product, ALK. Here, we show molecular cloning of cDNAs for both the human and mouse ALK proteins. The deduced amino acid sequences reveal that ALK is a novel receptor protein-tyrosine kinase having a putative transmembrane domain and an extracellular domain. These sequences are absent in the product of the transforming npm-alk gene. ALK shows the greatest sequence similarity to LTK (leukocyte tyrosine kinase) whose biological function is presently unknown. RNA blot hybridization analysis of various tissues reveals that the alk mRNA is dominantly detected in the brain and spinal cord. Immunoblotting with anti-ALK antibody shows that ALK is highly expressed in the neonatal brain. Furthermore, RNA in situ hybridization analysis shows that the alk mRNA is dominantly expressed in neurons in specific regions of the nervous system such as the thalamus, mid-brain, olfactory bulb, and ganglia of embryonic and neonatal mice. These data suggest that ALK plays an important role(s) in the development of the brain and exerts its effects on specific neurons in the nervous system.

[Constituents of the petroleum ether and ethyl acetate extract fractions from Dracaena cochinensis (Lour.) S.C. Chen].

Six constituents have been isolated from the petroleum ether and ethyl acetate extract fractions of Dracaena cochinensis. Their structures have been identified as 1,2,4,5-tetrachloro-3,6- dimethoxy-benzene,docosyl alcohol, octadecyl acetate, eicosyl acetate, resveratrol and 4',7-dihydroxy-flavone on the basis of physical, chemical and spectral deta. Of these compounds 1,2,4,5-tetrachloro-3,6-dimethoxy-benzene is a new one.

Structural and functional aspects of the multiplicity of Neu differentiation factors.

We used molecular cloning and functional analyses to extend the family of Neu differentiation factors (NDFs) and to explore the biochemical activity of different NDF isoforms. Exhaustive cloning revealed the existence of six distinct fibroblastic pro-NDFs, whose basic transmembrane structure includes an immunoglobulin-like motif and an epidermal growth factor (EGF)-like domain. Structural variation is confined to three domains: the C-terminal portion of the EGF-like domain (isoforms alpha and beta), the adjacent juxtamembrane stretch (isoforms 1 to 4), and the variable-length cytoplasmic domain (isoforms a, b, and c). Only certain combinations of the variable domains exist, and they display partial tissue specificity in their expression: pro-NDF-alpha 2 is the predominant form in mesenchymal cells, whereas pro-NDF-beta 1 is the major neuronal isoform. Only the transmembrane isoforms were glycosylated and secreted as biologically active 44-kDa glycoproteins, implying that the transmembrane domain functions as an internal signal peptide. Extensive glycosylation precedes proteolytic cleavage of pro-NDF but has no effect on receptor binding. By contrast, the EGF-like domain fully retains receptor binding activity when expressed separately, but its beta-type C terminus displays higher affinity than alpha-type NDFs. Likewise, structural heterogeneity of the cytoplasmic tails may determine isoform-specific rate of pro-NDF processing. Taken together, these results suggest that different NDF isoforms are generated by alternative splicing and perform distinct tissue-specific functions.

Application of invasive microwave hyperthermia for the treatment of gliomas.

Twenty five cases of gliomas of the brain were operated upon by debulking the tumour masses. Following this, microwave hyperthermia was given by heating a measured volume of Ringer's solution instilled into the tumour cavity. This was followed by a 'dry treatment' without Ringer's solution. The follow up of these cases revealed that 11 cases have died and 14 cases are alive post-operatively. For those that are alive, the follow up period ranges from 21 to 41 months with the mean survival period of 31.1 months; in this group, 12 cases have a Kanofsky scale of 80-100, i.e. they are fully independent. The other two cases have a score of 50 or under and they need institutional care. In this study, we believe that the first order effect of microwave hyperthermia is predominantly thermal and in the published literature, and in this investigation, there is no clear evidence that microwave radiation produces any other beneficial and quantifiable effect on the tissue.