[Rationality evaluation of Shuanghuanglian Injection combined with Ampicillin Sodium for Injection based on sequential analysis].

The lack of rationality evaluation method for drug combination has long restricted its clinical application. In view of this, this study took Shuanghuanglian Injection as model drug and established a "physical-chemical-biological" sequential analysis method, which is expected to provide clues for improving the safety and effectiveness of clinical drug combination. With the methods of insoluble particle testing, isothermal titration calorimetry(ITC), and real time cellular analysis(RTCA), the rationality of Shuanghuanglian Injection combined with Ampicillin Sodium for Injection was assessed. The results showed that the number of insoluble particles>10 µm in the solution of the combination met the standard of Chinese Pharmacopoeia, while the number of insoluble particles>25 µm did not meet the standard. ITC detection demonstrated that the change of Gibbs free energy(ΔG) was less than 0 during the fusion process, indicating that the process was spontaneous and enthalpy-driven reaction. Therefore, the interaction between the two was mainly chemical reaction, and the internal substances may change. RTCA found that Shuanghuanglian Injection alone and Ampicillin Sodium for Injection alone basically had no inhibitory effect on the growth of HEK293 T cells, while the combination of the two suppressed the growth of HEK293 T cells, suggesting that the combination was toxic to HEK293 T cells. This study showed that Shuanghuanglian Injection and Ampicillin Sodium for Injection reacted, yielding toxicity. This suggested that the two should not be combined for application. With the "physical-chemical-biological" sequential analysis, the molecular interaction of drugs was clarified. The method can be further applied for evaluating the rationality of other Chinese and western medicine injections.

[Exploration on rationality evaluation approach of drug combination medication based on sequential analysis and machine learning].

Drug combination is a common clinical phenomenon. However, the scientific implementation of drug combination is li-mited by the weak rational evaluation that reflects its clinical characteristics. In order to break through the limitations of existing evaluation tools, examining drug-to-drug and drug-to-target action characteristics is proposed from the physical, chemical and biological perspectives, combining clinical multicenter case resources, domestic and international drug interaction public facilities with the aim of discovering the common rules of drug combination. Machine learning technology is employed to build a system for evaluating and predicting the rationality of clinical drug combinations based on "drug characteristics-repository information-artificial intelligence" strategy, which will be debugged and validated in multi-center clinical practice, with a view to providing new ideas and technical references for the safety and efficacy of clinical drug use.

Antibacterial constituents from Melodinus suaveolens.

To investigate the non-alkaloidal chemical constituents of the stems and leaves of Melodinus suaveolens and their antibacterial activities. Compounds were isolated and purified by repeated silica gel, Sephadex LH-20, RP18, and preparative HPLC. Their structures were elucidated by comparison with published spectroscopic data, as well as on the basis of extensive spectroscopic analysis. The antibacterial screening assays were performed by the dilution method. Fourteen compounds were isolated, and identified as lycopersene (1), betulinic aldehyde (2), 3ß-acetoxy-22,23,24,25,26,27-hexanordammaran-20-one (3), 3a-acetyl-2, 3, 5-trimethyl-7a-hydroxy-5-(4,8,12-trimethyl-tridecanyl)-1,3a,5,6,7,7a-hexahydro-4-oxainden-1-one (4), 3ß-hydroxy-28-norlup-20(29)-ene-17ß-hydroperoxide (5), 3ß-hydroxy-28-norlup-20(29)-ene-17α-hydroperoxide (6), ß-sitosterol (7), 28-nor-urs-12-ene-3ß, 17ß-diol (8), α-amyrin (9), ergosta-4,6,8(14),22-tetraen-3-one (10), 3ß-hydroxy-urs-11-en-28,13ß-olide (11), betulin (12), obtusalin (13), and ursolic acid (14). Among the isolates, compounds 1, 2, 6, 8, 10, and 14 showed potent antibacterial activities against the four bacteria. This is the first report of the antibacterial activity of the constituents of Melodinus suaveolens.

Antibacterial prenylbenzoic acid derivatives from Anodendron formicinum.

A phytochemical investigation on the stems of Anodendron formicinum led to the isolation of eight prenylbenzoic acid derivatives. Three of these were new compounds, designated as formicinuosides A (1), B (2), and C (3). Their structures were elucidated on the basis of extensive spectroscopic analysis, as well as by comparison with the reported spectroscopic data. This is the first report of chemical constituents from A. formicinum and their antimicrobial activities. Among the isolated compounds, compounds 4, 6 and 8 showed significant antibacterial activities against Providensia smartii with MIC values of 0.781 µg/mL. Moreover, compound 8 showed remarkable antibacterial activity against Escherichia coli with MIC value of 0.781 µg/mL.

Promotion of immunity of mice to Pasteurella multocida and hog cholera vaccine by pig interleukin-6 gene and CpG motifs.

A novel oligodeoxynuleotides containing 11 CpG motifs was synthesized and inserted into the VR1020 plasmid containing pig interleukin-6 (IL-6) gene (VPIL6) to construct recombinant plasmid, VPIL6C. The chitosan nanoparticles (CNP) were prepared by ionic cross linkage to entrap the VPIL6C (VPIL6C-CNP), VPIL6 (VPIL6-CNP) and CpG (CpG-CNP). 42-Day old female mice were divided into four groups and intramuscularly injected respectively with 6 pmol VPIL6C-CNP, VPIL6-CNP, CpG-CNP and VR1020-CNP along with the bivalent vaccines against the Pasteurellosis and hog cholera. The blood was weekly collected from mice after vaccination to detect the changes of immunoglobulins, specific antibodies, IL-2, IL-4, IL-6 and immune cells. 28 days after vaccination, the mice were orally challenged with virulent Pasteurella multocida. The results showed that in comparison with those of the control VR1020 group, the content of immunoglobulins, specific antibodies and interleukins significantly increased in the sera from the treated two groups (P<0.05). Meanwhile, the number of lymphocytes and monocytes also remarkably elevated in the treated groups (P<0.05). The immune responses of VPIL6C mice were notably stronger than those of VPIL6 and CpG group. The challenge results proved that the overall immunity was further promoted in the treated mice which resisted the challenge infection; while the control mice manifested evident symptoms and lesions, and died of infection. These suggested that VPIL6C-CNP could better promote the immunity and resistance of mice against Pasteurellosis than VPIL6-CNP and CpG-CNP, and facilitate the development of effective adjuvant to enhance the immunity of animal against infection.

Enhancement of immunity to an Escherichia coli vaccine in mice orally inoculated with a fusion gene encoding porcine interleukin 4 and 6.

Experiments were conducted to investigate the effect of a fusion gene of porcine IL-4 and IL-6 (PIL4/IL6) packaged with chitosan nanoparticles (CNPs) in terms of the development of a novel effective adjuvant. The IL4/PIL6 fusion gene was constructed and inserted into a eukaryotic expression vector. The plasmid was bound to CNP and then utilized to orally inoculate 21-day-old female Kunming mice that simultaneously received intramuscular injection of inactivated Escherichia coli vaccine. At 35 days post-vaccination, the mice were challenged by oral feeding with virulent O139: K88 strain EPEC E. coli bacteria. Compared with those of control mice, the content of immunoglobulins and specific antibodies to E. coli increased significantly in the sera of mice immunized with VPIL4/IL6-CNP (P<0.05). Furthermore, the levels of IL-2, IL-4 and IL-6 increased remarkably in the sera of immunized mice (P<0.05). After challenge, these immunological markers were elevated to different degrees in the mice immunized with the fusion gene construct (IL4/VPIL6-CNP) or individual plasmids (VPIL4+VPIL6-CNP). The immunized mice all survived the challenge and did not show any symptoms or lesion, whereas the VR1020-CNP control mice manifested obvious clinical symptoms and hemorrhagic lesions in the digestive tracts. These results demonstrated that VPIL4/IL6 entrapped with CNP is a novel promising adjuvant to promote specific immunity and resistance of animals against infectious pathogen.