A large amount of data generated in the study of chromatographic fingerprint of Chinese herbal medicines. Criteria selection was one of main considerations while the fingerprint analysis was conducted. Based on the published reports, several criteria of chromatographic fingerprint analysis were reviewed in this paper, including the basic parameters, criteria of peaks isolated, reproducibility of chromatogram and related criteria,criteria of optimization, criteria of stability as well as multi-phase multi-information fingerprint chromatograms, etc..

Objective To observe the effect of severe heatstroke on intestinal mucosal barrier function,and explore its correlation with systemic inflammatory reaction.Methods The SPF male BALB/c mice were randomly divided into normal control group,40 ℃ heat stress group and 42 ℃ heat stress group,with 6 mice in each group.The mice in normal control group were observed at normal temperature with (25.0 ± 0.5)℃,and the mice in heat stress groups were challenged with a temperature of (35.5 ± 0.5) ℃ and a humidity of (60 ± 5)％ until body temperature increase up to 40 ℃ or 42 ℃ followed by recovering the surroundings temperature to normal temperature for 12 hours.The blood of medial angle of eye of mice in each group was collected for determination of plasma lipopolysaccharide (LPS),tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) levels,and diamine oxidase (DAO) activity with enzyme linked immunosorbent assay (ELISA).The level of D-lactic acid was determined with ultraviolet spectrophotometer.Then the mice in each group were sacrificed,and mesenteric lymph node (MLN),liver,spleen,lung,kidney tissues,and the blood from portal vein and caval vein were collected for colony count to observe the intestinal bacterial translocation.The ileum tissue was collected for observation of changes in histomorphology and ultrastructure of small intestine mucous membrane with microscope.Pearson linear regression analysis was used to explore the correlation between intestinal mucosal barrier dysfunction and systemic inflammatory response.Results Compared with normal control group,plasma LPS,inflammatory parameters such as TNF-α and IL-6,and gut barrier function parameters such as DAO and D-lactic acid levels as well as the rate of bacterial translocation after heat stress were significantly increased,and the differences were more obvious in 42 ℃ heat stress group [LPS (EU/L):740±50 vs.340±40,TNF-α (ng/L):148.06±36.61 vs.12.89 ± 1.67,IL-6 (ng/L):110.91 ± 9.97 vs.18.02 ± 2.20,DAO (U/L):1 760 ± 400 vs.670± 50,D-lactic acid (mg/L):9.60 ± 1.48 vs.5.08 ± 0.28,rate of bacterial translocation:78.6％ (33/42) vs.9.5％ (4/42),all P ＜ 0.01].It was shown by Pearson linear regression analysis that plasma LPS,TNF-α,IL-6 were positively correlated with DAO activity (r values were 0.834,0.808,0.836,respectively) and D-lactic acid (r values were 0.811,0.811,0.800,respectively) in 42 ℃ heat stress group (all P =0.000).It was showed by microscope that the changes in histomorphology and ultrastructure changes in intestinal mucosa were found after heat stress,and was obvious in 42 ℃ heat stress group as following:villus atrophy and falling off,infiltration of neutrophils and lymphocytes,the microvillus on the surface of mucosa cells were short and small,arranged in disorder,the tight junction between epithelial cells became widen,the mitochondrion and endoplasmic reticulum swelled obviously.Conclusion During the early stage of severe heatstroke,the damage of intestinal mucosal was obvious,and it has close correlation with systemic inflammatory response.

Objective To observe the effect of heat stress-induced reactive oxygen species (ROS) burst on the regulation of expression of Bcl-2 and Bax in human umbilical vein endothelial cell (HUVEC) apoptosis induced by heat stress,and explore the pathogenesis of vascular endothelial damage caused by severe heat stroke.Methods HUVEC heat stress model was reproduced.Cells of heat stress group were incubated at either 39,41,or 43 ℃ for 2 hours,then all the cells were further incubated at 37 ℃ and 5％ CO2 for 24 hours.Before heat stress,cells of 43 ℃ heat stress group were pretreated with 10 μmol/L MnTMPyP,which was a specific scavenger of ROS,for 1 hour.Cells of control group were incubated at 37 ℃ and 5％ CO2.The amount of ROS was assayed with 2',7'-dichlorofluorescin diacetate (DCFH-DA) and dihydroethidium (DHE) staining.Apoptosis was determined by using staining with Hoechst33258.The mRNA expressions of Bcl-2 and Bax were determined by reverse transcription-polymerase chain reaction (RT-PCR).The protein levels of Bcl-2,Bax,caspase-3 were analyzed by Western Blot.In addition,the effect of MnTMPyP on heat stress-induced apoptosis was also studied.Results Compared with control group,there was no obvious change in cells after 39 ℃ heat stress.With the increase in heat stress temperature up to 41 ℃ and 43 ℃,viability of cells showed a lowering trend,with a burst of ROS,and an increase of mRNA and protein of Bax,and the protein of caspase-3 was significantly increased,the mRNA and protein of Bcl-2 were significantly decreased in a temperature-dependent manner.These changes were marked in 43 ℃ heat stress group as compared with those of the control group [cell viability:(46.00 ±4.00)％ vs.(96.33 ± 1.53)％,t=20.164,P=0.001; ROS (fluorescence relative value):400.67 ± 12.10 vs.99.33 ±4.04,t=32.909,P=0.001; Bax mRNA (A value):3.03 ±0.15 vs.1.00 ± 0.00,t=23.056,P=0.001; Bax protein (gray value):3.97 ±0.21 vs.1.00 ± 0.00,t=24.684,P=0.001; caspase-3 protein (gray value):4.80 ± 0.20 vs.1.00 ± 0.00,t=32.909,P=0.001 ; Bcl-2 mRNA(A value):0.42 ± 0.30 vs.1.00 ± 0.00,t=33.072,P=0.001 ; Bcl-2 protein (gray value):0.39 ± 0.25 vs.1.00 ± 0.00,t=42.212,P=0.001].It was shown that pre-condition with the antioxidant MnTMPyP significantly decreased the heat stress-induced expression of Bax,caspase-3,and apoptosis,and the expression of Bcl-2 was elevated [Bax mRNA (A value):2.00 ± 0.20 vs.3.33 ±0.25,t=7.184,P=0.002; Bax protein (gray value):2.03 ±0.25 vs.3.23 ±0.25,t=5.840,P=0.004; caspase-3 protein (gray value):2.07 ± 0.21 vs.5.00 ± 0.20,t=17.600,P=0.001 ; Bcl-2 mRNA(A value):0.71 ± 0.40 vs.0.42 ± 0.26,t=8.126,P=0.002; Bcl-2 protein (gray value):0.57 ± 0.31 vs.0.40 ± 0.06,t=5.091,P=0.007].Conclusions A burst in an increase of ROS plays an important role on heat stress-induced HUVEC apoptosis,and the mechanism is probably related to the expressions of Bcl-2 and Bax.The vascular endothelial cells apoptosis may be one of the pathogenetic factor in severe heat stroke.

Objective To observe the expressions of cytochrome C,Caspase-9,Caspase-3 and their relationships,and investigate apoptosis signal transduction mechanism after heat stress-induction in human umbilical vein endothelial cell (HUVEC),and explore pathogenesis of vascular endothelial damage in the wake of severe heat stroke.Methods Human umbilical vein endothelial cell heat stress model was set up.Control group were incubated at 37℃,while heat stress group of cells were incubated at 39℃,41℃,and 43℃ for 2h,then all the cells were further incubated at 37℃ for 24 h.Mitochondria of human umbilical vein endothelial cell were examined with electron microscopy.Apoptosis was analyzed by flow cytometry using Annexin V-FITC/PI staining,and protein levels of cytochrome C,caspase-9,caspase-3 were analyzed by western blot.Results In the control group (37℃),the structure of mitochondrial was intact in HUVEC under transmission electron microscope.In contrast,mitochondrial swelling was found in the group of 43℃ heat stress.Compared with control group,as increasing in heat stress temperature,the rates of induced apoptosis were 17.8％ at 41℃ and 25.6％ at 43℃,and the levels of cytochrome C,Caspase-9,and caspase-3 were significantly increased (P ＜0.05).There was no obvious change at 39℃ heat stress (P ＞ 0.05).Conclusions Mitochondrial apoptosis pathway is involved in apoptosis of human umbilical vein endothelial cells in the wake of heat stress.The vascular endothelial cells apoptosis may be associated with the occurrence of severe heat stroke.

Results With the prolonged exposure to heat , the mice exhibited swollen and disorderly arranged neurons , shrunken cells , and contracted and deeply stained nuclei , with significantly higher scores on nerve pathological injury evaluation at 6, 12, and 24 h (2.78 ± 0.71, 3.21 ±0.56, and 3.36 ±0.63) than the control mice (0.43 ±0.10) (P<0.05).ELISA showed remarkably elevated levels of UCH-L1 in the serum (F=147.7, P=0.05) and brain tissue (F=145.7, P=0.05) in the heat stress group as compared with the con-trol, and Western blot also revealed a markedly higher expression of UCH-L1 in the brain tissue in the former group than in the latterObjective The abnormal expression of ubiquitin C-terminal hydrolase-L1 ( UCH-L1 ) has an important role in the diagnosis and prognosis of brain damage .This study was to investigate the changes of UCH-L1 in the serum and brain tissue in the mouse model of heat stress . Methods Twelve BALB/c mice were randomly divided into a control and a heat stress group of equal number, the former placed at a temperature of (25.0 ±0.5)℃and a relative humidity of (35 ±5)%and the latter in a simulated in-cubator at (35.5 ±0.5)℃and a relative humidity of (60 ±5)%.When the rectal temperature reached 42℃, the animals were re-moved from the incubator and cooled at an ambient temperature of (25.0 ±0.5)℃and a humidity of (35 ±5)%for 0, 6, 12, and 24 h.Then the brain tissues of all the animals were harvested for HE staining , evaluation of neuronal injury under the light microscope , measurement of the UCH-L1 levels in the serum and brain tissue by ELISA , Western blot, and immunohistochemistry , respectively. (F=261.2, P=0.01).Immunohistochemistry manifested that , with the prolonged exposure to heat , the UCH-L1 expression in the brain tissue was characterized by gradually increased light brown of the neurons at staining . Conclusion Severe heatstroke causes brain injury in a time-dependent manner , and the abnormally elevated levels of UCH-L1 in the serum and brain tissue can be a marker of heatstroke-induced brain injury .

Objective To explore the effect of activation of caspase-3 on apoptosis of human umbilical vein endothelial cells induced by heat stress. Methods A model of human umbilical vein endothelial cell heat stress was established. The endothelial cells in the control group were incubated at 37 ℃, while the cells in the heat stress group were incubated at 39 ℃, 41 ℃, or 43 ℃ for 2 h , then all the cells were further incubated at 37 ℃ for 24 h. Apoptosis was detected using staining with Hoechst33258; protein levels of caspase-3 were analyzed by Western blot; and the effect of caspase-3 inhibitor Z-DEVD-FMK on heat stress-induced apoptosis was determined. Results As compared with the control group, with an elevation in heat stress temperature (41℃ to 43 ℃), apoptosis was markedly increased and level of caspase-3 was significantly increased. In addition , caspase-3 inhibitor Z-DEVD-FMK significantly decreased the heat stress-induced apoptosis and levels of caspase-3. Conclusions Caspase-3 mediates apoptosis of human umbilical vein endothelial cells can be induced by heat stress.

Rhubarb is a traditional Chinese medicines which possess laxative, lipid-lowering, and weight-loss activities, but the active compounds of lipid-lowering and underlying molecular mechanisms are not yet clear. This study aims to explore the effects of chrysophanol on the mRNA expressions of sterol regulatory element-binding proteins (SREBPs) and lipid metabolism in human liver carcinoma Huh-7 cells, which is one of the active compounds obtained from Rhubarb. A reporter gene assay was used to test the transcription of SREBP. The intracellular triglyceride and total cholesterol contents were measured by using commercially available test kits. The SREBPs target genes expressions were measured by Quantitative Real-Time PCR. Cell viability was evaluated by Cell Counting Kit-8. As the results shown, chrysophanol (40 μmol · L(-1), 16 h) could notably inhibited human SRE promoter activity in a dose-dependent manner and decrease intracellular cholesterol and triglyceride levels. Furthermore, the mRNA expressions of SREBPs target genes were significantly downregulated by chrysophanol treatment. However there are no significant differences on cell viability when compared with the control group. These results suggested that chrysophanol might improve lipid metabolism through suppressing the mRNA expressions of SREBPs target genes to attenuate intracellular lipid accumulation.

Objective To study the benzofuran compounds from roots and rhizomes of Ligularia caloxantha, which is a folk medicine used in the Naxi Nationality in Yunnan Province. Methods Compounds were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were elucidated by physicochemical properties and spectral analysis. Results Eighit compounds are isolated from ethanolic extracts of the roots and rhizomes. They were identified as euparin (Ⅰ), 6-methoxy-euparin (Ⅱ), 4, 5-dimethoxy-2-hydroxybenzaldehyde (Ⅲ), 2-acetyl-5, 6-dimethyoxybenzofuran (Ⅳ), 4-hydroxyacetophenone (Ⅴ), 2-isopropenyl-5, 6-dimethoxy-2, 3-hydrocumaran (Ⅵ), 8?-hydroxy-7(11)-eremophilen-12, 8?-olide (Ⅶ), lupeol (Ⅷ). Conclusion All the eight compounds were obtained from L. caloxantha for the first time as benzofurans. Compounds Ⅰ and Ⅱ are two major ones with insecticide and insect food refusal-induced activities. That is the relative reason of L. caloxantha used for folk anti-insect.

The medicinal herbs derived from genus Senecio have been commonly used in Chinese medicine and triggered attention in recent decades for that they contain the hepatotoxic pyrrolizidine alkaloids. Therefore the botanical pharmacognostic study to authenticate those herbs based on their macroscopic and microscopic characteristics is important for the assurance of safety when they are applied as raw material for extracts or for finished products. In this paper, 13 taxa (11 species and 2 varieties) of Senecio plants were collected and their macroscopic and microscopic characteristics were observed and described by digital microscopic illustration. The results showed that the distribution of collenchyma in the cortex, the level of development for pericycle, the location of the phloem, and the ratio of pith in transverse sections of the stems, and the morphology of the leaf epidermal cells, the stomatal types and the non-glandular hairs in leaf surface view were found to be the main microscopic characteristics for authentication of different Senecio species. The herbs derived from genus Senecio can be distinguished from each other on the basis of their macroscopic and microscopic characteristics, and those observation can be used for the identification of commercial crude drugs from Senecio plants.

Danshen (Salvia miltiorrhiza Bunge) hairy roots were obtained by infecting Danshen leaves with Agrobacterium rhizogenes 9402. Besides rosmarinic acid (RA) and salvianolic acid B (SAB), the hairy root could also produce salvianolic acid K (SAK), salvianolic acid L, ethyl salvianolic acid B (ESAB), methyl salvianolic acid B (MSAB), and a compound with a molecular weight of 538 (compound 538) identified by using LC-MS. Effects of methyl jasmonate (MeJA) and yeast elicitor (YE) on the accumulation of these compounds had been investigated. MeJA increased the accumulation of SAB, RA, SAK, and compound 538 from 4.21%, 2.48%, 0.29%, and 0.01% of dry weight to 7.11%, 3.38%, 0.68%, and 0.04%, respectively. YE stimulated the biosynthesis of RA from 2.83% to 5.71%, but depressed the synthesis of SAB, SAK and compound 538. It was indicated in all the results that these Danshen hairy roots could be used as alternative resources to produce salvianolic acids. Analysis of the content variation of these compounds after elicitation suggested that SAK and compound 538 might be the intermediates in the biosynthesis from RA to SAB in Danshen hairy roots.