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【Objective】 To explore the potential molecular biological mechanism of Belamcanda chinensis in the treatment of glioma based on network pharmacology, molecular docking technology and in vitro cell experiments. 【Methods】 ① The active components, targets of Belamcanda chinensis and targets of glioma were obtained by database search. String database was used to analyze protein-protein interaction relationship, R project was used to analyze gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, Cytoscape software was used to build "compound-target-disease" network and PPI network, and AutoDock software was used to verify molecular docking. ② Western blotting, qRT-PCT and apoptosis assay were used to verify the enrichment results of network pharmacology targets and protein pathway. 【Results】 ① We screened out 32 types of active components, 484 types of targets and 464 types of glioma targets, and obtained 62 kinds of therapeutic targets after mapping. We obtained 12 kinds of key pharmacodynamic molecules such as Isoiridogermanal, Iridobelamal A and Rhamnazinand and other key pharmacodynamic molecules, as well as AKT1, STAT3, HRAS and other core targets by network topology analysis. Enrichment analysis results demonstrated that they were mainly involved in biological processes such as peptide serine phosphorylation, protein kinase B signal transduction, peptide serine modification, and pathways including PI3K/AKT signal pathway and Rap1 signal pathway. The results of molecular docking verified the good binding activity of the key pharmacodynamic molecules with the core targets. ② The results of Western blotting showed that the protein expressions of VEGF and MMP9 of Belamcanda chinensis extracts in 8 mg/mL and 16 mg/mL groups were significantly lower than those in the blank control group (P<0.01 or P<0.001). Compared with the blank control group, the early apoptosis rate of Belamcanda chinensis extracts at 8 mg/mL and 16 mg/mL were significantly decreased (P<0.001 or P<0.000 1). qRT-PCR results showed that the mRNA expression levels of VEGF and MMP9 in Belamcanda chinensis extracts at 8 mg/mL and 16 mg/mL were significantly decreased (P<0.001 or P<0.0001). 【Conclusion】 The treatment of glioma with Belamcanda chinensis is the result of multi-component, multi-target and multi-channel interactions. The results of cell experiments confirmed that Belamcanda chinensis extracts can affect the expressions of related target proteins of PI3K/AKT signal pathway and VEGF and MMP9, which verified the results of network pharmacology. The results provide a theoretical basis for the clinical application of Belamcanda chinensis and studies on glioma.
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Understanding the effect of immunosuppressive agents on intestinal microbiota is important to reduce the mortality and morbidity from orthotopic liver transplantation (OLT). We investigated the relationship between the commonly used immunosuppressive agent cyclosporine A (CSA) and the intestinal microbial variation in an OLT model. The rat samples were divided as follows: (1) N group (normal control); (2) I group (isograft LT, Brown Norway [BN] rat to BN); (3) R group (allograft LT, Lewis to BN rat); and (4) CSA group (R group treated with CSA). The intestinal microbiota was assayed by denaturing gradient gel electrophoresis profiles and by using real-time polymerase chain reaction. The liver histopathology and the alanine/aspartate aminotransferase ratio after LT were both ameliorated by CSA. In the CSA group, the numbers of rDNA gene copies of Clostridium cluster I, Clostridium cluster XIV, and Enterobacteriaceae decreased, whereas those of Faecalibacterium prausnitzii increased compared with the R group. Cluster analysis indicated that the samples from the N, I, and CSA groups were clustered, whereas the other clusters contained the samples from the R group. Hence, CSA ameliorates hepatic graft injury and partially restores gut microbiota following LT, and these may benefit hepatic graft rejection.
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Objective To explore the clinical application of combined cooling of Chinese and Western medicine on the cooling effect of patients with heatstroke. Methods A total of 80 patients with severe heat stroke admitted to the emergency department of our hospital from January 2016 to December 2017 were selected and divided into treatment group and control group according to the random number table method. 40 patients in the control group were given physical cooling. For conventional treatment, patients in the treatment group were given TCM treatment interventions on the basis of this, and clinical efficacy, changes in body temperature, and adverse screening events were compared between the two groups. Results The clinical effective rate (100.00%, ) in the treatment group was significantly higher than that in the control group (85.00%, Z=6.125,P<0.05). The duration of high fever, duration of fever, and recovery time of symptoms and signs were (0.57±0.46), (3.12±0.75), (7.25±3.18) h in the treatment group, (1.43±0.89), (5.37±0.63), (12.47± 4.53) h in the control group, there were significant differences between the two groups (t=5.429, 14.528, 5.965, P<0.05). After treatment, the levels of cTnl and CK-MB in the peripheral blood of the two groups decreased significantly (P<0.05). The levels of cTnl and CK-MB in peripheral blood of patients in the treatment group were (3.13±0.15) μg/L, (412.02±156.33) U/L, (3.54± 0.26) μg/L, (748.32±119.20) U/L in the control group, there were significant differences between the two groups (t=8.639, 10.819,P<0.05).There was no significant difference in the incidence of adverse reactions between the two groups (χ2=2.222, P=0.263). Conclusion The combination of Chinese and Western medicine physical cooling has a significant cooling effect on patients with heat stroke, and does not increase the risk of adverse reactions, it is worth clinical Promotion and application.
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@#Objective To investigate the immediate effects of repetitive peripheral magnetic stimulation (rPMS) on upper limb spasticity and motor function for stroke patients. Methods From May to October, 2018, 14 stroke patients accepted rPMS once. They were assessed with modified Ashworth Scale (MAS) and modified Tardieu Scale (MTS) of Upper limb, and Fugl-Meyer Assessment-Upper Extremities (FMA-UE) before and after stimulation. Results The score of FMA-UE impoved (t = -3.166, P < 0.01) after rPMS, as well as those of MAS of shoulder adductors, shoulder extensors, elbow flexors, elbow extensors and wrist flexors (P < 0.05), and R1 of shoulder adductors, shoulder extensors, elbow flexors, elbow extensors and wrist flexors in MTS (P < 0.05), R2 of shoulder adductors and shoulder extensors (P < 0.05). Conclusion rPMS may immediately effect spasticity and motor function on upper limbs in stroke patients.
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Objective To study the cause,prevention and treatment of postpancreaticoduodenectomy hemorrhage.Method The clinical data of 422 patients who underwent PD in our hospital between January 2000 and January 2012 were retrospective analyzed.Results The incidence of postoperative hemorrhage was 8.1％ (34/422),and the mortality was 20.6％ (7/34).Early and delayed hemorrhage occurred in 19 and 15 patients,respectively.Intra-abdominal and gastrointestinal hemorrhage occurred in 20 and 14 patients,respectively.For the 19 patients who underwent reoperation,the mortality was 20.6％ (7/34).When compared with the delayed hemorrhage group,the mortality of the early hemorrhage group was significantly lower (P＜0.05).Conclusions Meticulous operation and reliable hemostasis during operation and prevention of pancreatic fistula,biliary fistula and peritoneal fluid collection after operation are the key points in reducing postoperative hemorrhage.A timely and decisive reoperation is important to manage postoperative hemorrhage.
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Objective To analyze the diversity of intestinal microecology in different healthy age groups and explore the relationship between age and the composition of intestinal microflora.MethodsFecal samples were collected from the children,adult and elderly groups.The main intestinal microflora were cultured and quantified,and the qualitative and quantitative changes in the level of category were determined by fluorescence quantitative PCR.The Bifidobacteria/Enterobacteriaceae (B/E) ratio reflexing intestinal colonization resistance was also calculated.Results The germiculture of fecal samples indicated that the counts of Bifidobacteria and Lactobacillus (P＜0.01 and P＜0.05,respectively) were significantly lower in the adult and aged groups than in children group.The number of Bacteriodes and Enterobacteriaceae in the elderly group were higher than in the children and the adult groups (all P＜0.01).The counts of Enterococcus were increased in the adult and the aged groups (P＜0.05 and P＜0.01) compared with the children group,and were higher in the aged group than in the adult group (P＜0.05).The B/E ratio was significantly decreased in the aged group (0.88-±-0.13) as compared with the children (1.15±0.15) and the adult groups ( 1.01± 0.20) ( both P ＜ 0.01).The diversities of Bifidobacteria,Bacteriodes,Enterobacteriaceae and B/E ratio by fluorescent quantification PCR were consistent with the results by germiculture technology.The diversity of Lactobacillus (P＜0.01) between the children and the aged groups as well as between the children group and the adult group significantly improved (P＜0.01 and P＜0.05 ) in PCR compared with the germiculture technology. ConclusionsThe numbers of intestinal Bifidobacterium and Lactobacillus are decreased with lower colonization resistance,while the numbers of Bacteriodes,Enterobacteriaceae and Enterococcus are increased with aging.
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Objective To determine the changes of peripheral levels of T helper cell cytokines of patients with chronic hepatitis B (CHB) during antiviral treatment,and to further explore its clinical significance.Methods The plasma levels of interleukin (IL)-2,IL-6,IL-10,interferon (IFN)-γ and tumor necrosis factor(TNF)-α of thirty-three CHB patients during antiviral treatment (entecavir) were measured using enzyme linked immunosorbont assay (ELISA).And their biochemical indicators of liver function were determined.The differences of cytokines levels before and after antiviral treatment were compared using ANOVA.The correlations between the changes of cytokines and alanine transaminase (ALT),hepatitis B virus (HBV) DNA levels were analyzed.Results Levels of IFNγ before and 12,24,48 weeks after treatment were (5.98±2.77),(5.95±3.37),(2.93±2.15) and (9.29±4.65) pg/mL,respectively (F=3.845,P＜0.05),which were positively correlated with ALT levels (r =0.396,P＜0.05).Both TNF-α and IL-10 levels declined after antiviral treatment,which were significantly different at different time points (F=20.156 and 16.695,respectively; both P＜0.05),and both levels of TNF-α and IL-10 were positively correlated with ALT levels (r=0.354and 0.316,respectively; both P＜0.05) and positively correlated with HBV DNA levels (r=0.382and 0.386,respectively; both P＜0.05).While both IL-2 and IL-6 levels were not significantly different between before and after antiviral treatment (F=0.010 and 0.932,respectively; both P＞0.05).The serum levels of ALT and HBV DNA before and after antiviral treatment were all significantly different (F=17.69 and 198.98,respectively; both P＜0.05),which declined gradually during treatment and were positively correlated (r =0.581,P＜0.05).Conclusions IL-10,IFNγand TNF-α may be involved in the pathologic process of CHB,and closely related to the deterioration of the disease.Monitoring plasma levels of these cytokines during antiviral treatment could be useful to understand the immune status and evaluate the efficacy of antiviral drugs.
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Objective To investigate the effect of combined prism lenses for the treatment of juvenile myopes. Methods 622 cases of juvenile myopes ranging from - 1. 0D to - 5. 0D, aging from 6 - 18ages, were divided randomly into two groups. The control group(320 cases) wear general glass and the research group(302 cases) wear combined prism lenses. The juvenile myopes of both groups were checked their vision every 6 months and data were collected at the end of the fourth 6 months and analyzed by SPSS software. Results Ther eyes of increasing diopter in research group were 38(6.3%of all eyes) ,33 eyes were less than 1. 0D and 5 eyes were more than 1. 0D among them;but 294(46.0% of all eyes) in the control,201 eyes were less than 1.0D and 93 eyes were more than 1.0D among them. There were significant differences between two groups(P＜0.01). Conclusion Combined prism lenses for juvenile myopes (lower and middle) .compared with the common glass, had good clinical effect and it was a reliable and practicable method,to wear combined prism lenses for the treatment of juvenile myopes.
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Objective To analyze the association between mtDNA mutations and photodamagc after ultraviolet B (UVB) irradiation. Methods Primary human skin fibroblasts (HSF) and primary human epi- dermal keratinocytes of adult (HEKa) were irradiated by sub-lethal doses of UVB thrice a day for 4-5 days. Thereafter, genomic DNA was extracted from irradiated cells and conventional PCR was applied to detect the frequency rates of 4977 bp and 3895 bp mtDNA deletion. To quantitatively analyze the mutation levels, SYBR Green real-time PCR method was performed. Results In both cell lines, the frequency rates and relative copy number of deletions increased with the cumulative doses of UVB exposure (P<0.05). The prevalence rate of 3895 bp deletion peaked 53.3% and and relative copy number reached (49.63±4.38)×10-5, showing a more intense response to the accumulation of UVB radiation than 4977 bp deletion. In HSF, the minimum cumu- lative dose of UVB radiation was 150 mJ/cm2 for the induction of 3895 bp deletion, and 200 mJ/cm2 for the induction of 4977 bp deletion. It seemed that mtDNA deletion was more readily to be induced by UVB radia- tion in HSF than in HEKa. Conclusions The development and accumulation of mtDNA mutation are intimately related with cumulated UVB dose received by skin cells, and the 3895 bp deletion is more reliable in moni- toring the photodamage caused by UV than 4977 bp deletion. Therefore, the 3895 bp deletion may serve as a biomarker for the detection of photodamagc in skin cells. HSF appear to have an increased susceptibility to UVB radiation, which results in a higher frequency and level of mtDNA mutations compared with HEKa.
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Objective To investigate the ecological structure of intestinal fungal flora of patients with chronic hepatitis B infection. Methods HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers were selected as research subjects to extract the total DNA from stool of each subject, 18S rRNA genes of intestinal fungi was cloned and sequence was analyzed for the construction of 18S rDNA clone libraries and phylogenetic tree, diversity and structural characteristics of intestinal fungal flora in each group was then analyzed. Results A total of 29 operational taxonomic units (OTUs) of intestinal fungal flora in all research subjects was acquired. All positive clones belonged to three fungal taxa: Zygomycetes (3.4%) , Ascomycetes (82. 8%) and Basidiomycetes (13. 8%) . The dominant fungal community was Candida spp. , uncultured fungus and Saccharomyces spp. , which accounted for 29. 2% , 15. 9%, 15.0% respectively. The OTUs which belonged to HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers was 20,16,12,14 respectively. Conclusion There is an abundant fungal flora in the human intestine. Ecological structure of intestinal fungal flora appears to be various in patients with chronic hepatitis B infection, which indicate the close relationship between the alteration of ecological structure of fungal flora and the stage of chronic hepatitis B infection.