BACKGROUND: By targeting inducing differentiation in vitro,mesenchymal.stem cells(MSCs) transform into osteoblasts,lipocytes,chondrocytes,muscular cells,neuronal cells,etc. Whether Chinese herbs act on induced differentiation of MSCs in rats or not?OBJECTIVE: To study the amplification of MSCs cultured in vitro in SD rats and efficacy of Chinese herbs on targeting inducing differentiation of neuron-like cells.DESIGN: Exploring study with repeated observation and measurement based on cells.SETTING: Department of pathophysiology in a medical college.MATERIALS: Experimental marrow collected from SD male tested-healthy rats.METHODS: By adhesion method,MSCs in rats were isolated for amplifying culture in vitro. Flow-type cell instrument was applied for the determination of its surface antigen expression. Various Chinese herbal components were used for the targeting inducing differentiation of MSCs into neuron-like cells. The cellular morphology was observed under optical microscope. and the specific antigen label of neuronal cells was determined with immunocyto-chemical method.MAIN OUTCOME MEASURES:①Results of MSCs isolation and amplification;②Results of identification of MSCs surface antigen and neuon-like cells.RESULTS: By adhesion method,MSCs in rats were isolated successfully and amplified in a large amount in vitro. It was indicated in the results determined by flow-type cell instrument that CD14,CD1 1α,CD34,CD38,CD45,CD80 and CD86 presented negative,and CD29,CD44,CD90,CD105 and CD166 presented positive. By induced with various kinds of Chinese herbs,like huangqi(Radix Astragali seu Hedysari),tianma(Rhizoma Gastrodiae),renshen (Radix Ginseng),danggui(Radix Angelicae Sinensis),naoxinshu,renshen fengwangjian for 1 to 3 hours,most MSCs transformed into neuron-like cells,presenting soma and neurite. With immunocyto-chemical staining,neuron-specific enolase(NSE) and nestin displayed positive and glial fibrillary acidic protein negative.CONCLUSION: MSCs in SD rats have the potential of multi-targeting differentiation,presenting a strong capacity of amplification and self-replacement. In a suitable inducing condition,MSCs may differentiate into neuron-like cells.

OBJECTIVEThrough determination of the dynamic change of the active component in different parts of Prunella vulgaris at different growth stages, to find the optimal harvest time.

METHODTotal flavonoids content was determined by using the spectrophotometric method, and the content of ursolic acid and oleanolic acid was determined by HPLC. The contents of ash and extract were determined according to the methods in Chinese Pharmacopeia (2005 edition).

RESULTThere existed the active components in all parts of P. vulgaris, but the active component contents in different parts of P. vulgaris of at different growth stages, changed very obviously.

CONCLUSIONIn Yangtze-Huaihai region, the optimal harvest time of Prunella spike best harvest is at the end of June, and Prunellastem at the end of May. All parts of P. vulgaris have medicinal value.

Chromatography, High Pressure Liquid ; Flavonoids ; analysis ; Oleanolic Acid ; analysis ; Prunella ; chemistry ; Triterpenes ; analysis

Objective To establish an HPLC quantitative method for the content determination of baicalin in Kangdeling capsule .Methods The chromatography column was Agilent Tc-C18-WR (4 .6 mm × 250 mm ,5μm) ,and the column temper-ature was 30 ℃ .The mobile phase consisted of acetonitrile and 0 .5‰ phosphoric acid (26 ∶ 74 ) .The flow rate was 1 .0 ml/min ,and the detection wavelength was 265 nm .Results The retention time of baicalin was about 16 min .The calibra-tion equation was Y=22 114 .67 X -112 836 .7(r=0998 8)with good linearity in the range of 5 .410-108 .2 μg/ml for baicalin . The average recovery was 98 .78% while RSD were 0 .74% .Conclusion This method is simple ,time-saving and accurate which could be used to routine analysis of baicalin in Kangdeling capsule .

Objective To develop a method for determination of the rosmarinic acid in Xiaokang capsules .Methods An HPLC method was developed .The determination was performed on Agilent C18 column at 30℃ with a mobile phase composed of methanol :0 .5% formic acid (50∶50) run at a flow rate of 1 .0 ml/min .The detection wavelength was set at 330 nm and the injection volume was 10 μl .Results The linear range of rosmarinic acid concentrations was 2 .88 to 20 .1μg/ml (r=0 .999 8) . The average recovery was 99 .5% (RSD=0 .40% ,r=9) .Conclusion An assay for the determination of the rosmarinic acid in Xiaokang capsule that is simple ,rapid ,accurate and reliable has been developed .

Objective To detecte the differential metabolites and related pathways in Siha cells of cervical cancer by screening the inhibition of HPV16 E6/E7 expression based on 1H NMR metabolomics so as to identify the key metabolic markers involved in the development of high-risk HPV16 cervical cancer.Methods Siha cells were transfected with RNAi fragments to down-regulate the expression of E6/E7,which were divided into the normal control group(Siha cells),no-load group(si-NON),si-E6 group and si-E7 group,and their transfection efficiency was verified.1H NMR metabolomics was used to reveal the differential metabolites involved in interfering E6/E7 expression in Siha cells.Combined with MetaboAnalyst 5.0 online software,differential metabolites and related metabolic pathways were obtained.Results Fluorescence was observed by inverted fluorescence microscope.Western blotting results showed that compared with Siha group,the expression of E6/E7 in si-E6 group and si-E7 group was decreased(F=145.8,P<0.001).After down-regulating the expression of E6/E7,13 common differential metabolites,including Isoleucine,Leucine and valine,were detected.The results of MetaboAnalyst 5.0 online software analysis suggested that the above metabolites were mainly involved in the biochemical synthesis pathway of aminoacyl-trNA,biochemical synthesis pathway of isoleucine,Leucine and valine;There were 10 metabolic pathways of tyrosine,phenylalanine and tryptophan biochemical synthesis.Conclusion After HPV16 infection,changes of glucose and amino acid metabolism can promote the progression of cervical cancer,which provide a theoretical basis for the prevention and treatment of cervical cancer.

Objective:To describe the characteristics and analyze risk factors for surgical items count near-miss errors stemming from the self-incident reports of staff nurses from operating room, to reduce the risk of counting surgical items and prevent the occurrence of the relative adverse events.Methods:This was a retrospective study. Used the self-made checklist to retrospect the surgical items count errors, relative characteristics and reasons from the operating room nurses of Department of Anesthesiology, Shandong Provincial Hospital Affiliated to Shandong First Medical University reported from January 2017 to December 2021. Grey Relational Analysis was used to analyze and identify the risk elements.Results:A total of 98 surgical items count near-miss errors were reported by nurses.The unclear items were mainly classified into 6 categories, of which 52.04% (51/98) were disposable surgical items, 24.49% (24/98) were fine parts of surgical instruments, 14.28% (14/98) were implants, 5.10% (5/98) were electrosurgical instruments, 3.06% (3/98) were power systems, and 1.02% (1/98) were medical lasers; the disposable surgical items were the highest risk of surgical items count near-miss errors (non-standard behaviors of surgeons ξ 1=0.333); among the 9 risk factors, non-standard behaviors of surgeons ( r1 = 0.673), instrument nurses improper operation ( r4 = 0.691) and surgeons errors ( r2 = 0.693) were the most important influence factors. Conclusions:Analyzing the possible system risk factors resulting from the near-miss error could be a useful method for nurses to generate hierarchical risk-control strategies and improve surgical items count safety for patients. This com prerent the occurrence of adverse events.

Objective To investigate the role of bufalin(BU)in inhibiting M2-type macrophage-mediated colorec-tal cancer metastasis.Methods Human acute leukemia mononuclear cells(THP-1)were differentiated into M0 macrophages using phorbol ester induction(PMA)for 48 hours.The M0 macrophages were then treated with IL-4 and IL-13 medium.Surface markers and morphological changes were observed through ELISA,morphology,and RT-qPCR experiments.RT-PCR and ELISA experiments were conducted to detect the surface markers TGF-β and IL-10 of M2 macrophages.The secretion level of IL-6 in the supernatant of M2 macrophages and colorectal cancer cells HCT116 was compared using ELISA.Additionally,the effect of conditioned medium on colorectal cancer cell HCT116 was assessed through Transwell,Wound healing,RT-qPCR,and Western blot experiments.Subsequent-ly,bufalin was added to the conditioned medium and the changes in AKT/PI3K protein,migration,and epithelial-mesenchymal transition ability in HCT116 were observed using Western blot,Transwell,Wound healing and RT-qPCR experiments.Results THP-1 were successfully differentiated into M2 macrophages.The activation of AKT/PI3K protein in HCT116 cells was induced by the secretion of IL-6 from M2 macrophages,which in turn promoted the migration and epithelial-mesenchymal transition ability of the HCT116 cells.The migration and epithelial-mes-enchymal transition mediated by M2 macrophages in HCT116 cells were effectively inhibited by Bufalin.Conclu-sion The release of IL-6 from M2 macrophages activates the AKT/PI3K signaling pathway in colorectal cancer cells,thereby promoting their migration and epithelial-mesenchymal transition capacity.Moreover,bufalin exhibits inhibitory effects on this effect.