Chemical profiling of Zhi-Ke-Bao pills and its potential mechanism against cough by ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry and network pharmacology.

Zhi-Ke-Bao pills (ZKB), a traditional Chinese medicine preparation composed of 13 herbs, is generally used to treat cough caused by external wind cold, phlegm, etc in clinical applications, and it plays a core role in relieving cough caused by COVID-19 and influenza in China. Till now, the understanding of its chemical constituents was dramatically limited due to its chemical complexity, restricting its clinical application or development. In this work, a developed ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS) method, a targeted and non-targeted strategy and network pharmacology were used to comprehensively characterize the chemical compositions in ZKB and predict its mechanism against cough. A total of 164 compounds (148 targeted compounds and 16 non-targeted ones) were identified or tentatively characterized in ZKB, including 65 flavonoids, 25 alkaloids, 19 organic acids, 41 saponins, 9 coumarins, 2 phenylpropanoids, 2 anthraquinones, and 1 other types. Among them, 37 compounds were unambiguously identified by comparison to reference standards. Meanwhile, the fragmentation behaviors of five main chemical structure types were also summarized. 309 targets and two core signaling pathways of ZKB against cough were predicted by network pharmacology, including MAPK and PI3K-Akt signaling pathways. It was the first time to characterize the chemical compounds of ZKB and reveal its potential mechanism against cough, providing the material basis for further quality control or pharmacodynamic evaluation of ZKB.

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma.

Immunotherapy with tumor lysate-pulsed dendritic cells (DCs) is one of the breakthrough strategies used in the treatment of cancer. However, DC-based immunotherapies for osteosarcoma are limited. In the present study, preclinical studies of a C3H osteosarcoma mouse model (produced by subcutaneous injection of LM8 murine osteosarcoma cells) validated the concept that LM8 cell lysate-pulsed bone marrow-derived DCs may evoke a more potent immune response compared with DCs that have been matured using polyinosinic:polycytidylic acid (poly I:C). A cytotoxic T lymphocyte (CTL) response was established using two groups of C3H mice (n=9) with osteosarcoma; the treatment group consisted of LM8 cell lysate-pulsed DCs and the control group consisted of DCs matured using poly I:C. Each group was immunized with doses of 1×106 cells twice per week for 3 weeks. No difference in the expression of cluster of differentiation markers was identified in the two groups. DCs pulsed with LM8 cell lysate were associated with the increased induction of CTL activity. Serum interferon-γ levels were increased in mice that received DCs pulsed with LM8 cell lysate compared with that in the poly I:C-matured DC group (P<0.041). Serum interleukin-4 was decreased in the treatment group vs. the control group (P<0.033). A mixed lymphocyte reaction assay confirmed that LM8-DC immunotherapy may evoke a significant antigen-specific immune response in a mouse model. The present study reveals promising data on efficacy of a DC-based immunotherapy in the treatment of osteosarcoma; however, further clinical studies are warranted.

[Study on two different aromas styles of tobacco from Guizhou by characteristics spectroscopic methods].

This paper made use of three-dimensional fluorescence and ultraviolet-absorption spectrum to analyze the spectral characteristics of etroleum ether extract from Guizhou flue-cured tobacco and the overall characteristic spectral information of tobacco chemical substances were obtained. The three dimensional fluorescence and ultraviolet-visible absorption spectrum of each petroleum ether extract of flue-cured tobacco from different areas are generally similar, but their intensity is different. There have three characteristic peaks in three dimensional fluorescence spectra: I: Ex/Em = 297/326 nm, II: Ex/Em = 250/330 nm, III: Ex/Em = 225/336 nm respectively and meanwhile the order of these peaks intensity is I > III > II. The ultraviolet-visible absorption spectrum in 300-300 nm range presents four characteristic absorption peaks, whose maximum absorption wavelength are 329, 419, 445 and 419 nm respectively. Meanwhile, in accord with the relative intensity of characteristic peaks, it is known that there exist differences in the relative contents of the total chemical substances obtained from different flavor styles of the flue-cured tobacco. The clustering analysis results of three-dimensional fluorescence intensity score (D) and intensity ratio (R) show that in a certain range of distance coefficient, the flue-cured tobacco from different regions in Guizhou can be clearly divided into two classes "mildly sweet "and "alcohol sweet ". The classification can be well achieved in the smaller distance coefficient according to the ratio cluster of fluorescence intensity instead of the score cluster of fluorescence intensity. The method of three-dimensional fluorescence was better than that of ultraviolet-visible spectrometry in the matter of the clustering characteristic.

[Rehabilitation care for children after trauma in the earthquake disaster].

For the children who suffer trauma in earthquake, rehabilitation care aims to promote functional recovery, shorten hospital stay, and reduce the incidence of complications or disability by evidence-based, multidisciplinary, and comprehensive early rehabilitation intervention on the basis of first aid and clinical treatment. Children are likely to suffer traumatic brain injury, spinal cord injury, peripheral nerve injury, limb fracture, and amputation in the earthquake disaster, so the clinical rehabilitation care designed considering the characteristics of children should be provided immediately after acute phase of trauma to promote functional recovery.