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ObjectiveThe antitumor activity of sesquiterpenoid M36 isolated from Myrrha against human hepatoma HepG2 cells was investigated in this study. MethodHepG2 cells were treated with M36 at different concentrations （0， 2， 4， 6， 8， 10 μmol·L-1）. Firstly， the effects of M36 on the proliferation of human hepatoma HepG2 cells were detected by methyl thiazolyl tetrazolium （MTT）， colony formation assay， and EdU proliferation assay. Hoechst staining， flow cytometry analysis， and Western blot were used to explore the effect of M36 on the apoptosis of human hepatoma HepG2 cells. Acridine orange staining and western blotting were used to examine the effect of M36 on autophagy in HepG2 cells. Finally， Western blot was used to detect protein expression of cancer-related signaling pathways. ResultCompared with the blank group， M36 treatment significantly inhibited the proliferation of human hepatoma HepG2 cells （P<0.01）， and the half inhibitory concentration （IC50） value of M36 for 48 h was 5.03 μmol·L-1， in a dose- and time-dependent manner. M36 was also able to induce apoptosis and autophagy in human hepatoma HepG2 cells. After treatment with 8 μmol·L-1 M36 for 48 hours， the apoptosis rate of HepG2 cells was （42.03±9.65）% （P<0.01）. Compared with the blank group， HepG2 cells treated with 4 and 8 μmol·L-1 M36 for 48 h had a significant increase in cleaved poly ADP-ribose polymerase （cleaved-PARP） protein levels （P<0.01）. Acridine orange staining showed that autophagy was significantly activated in HepG2 cells treated with 4 and 8 μmol·L-1 M36 for 48 h compared with the blank group （P<0.01）， which was further verified by the up-regulation of microtubule-associated protein 1 light chain 3 Ⅱ （LC3 Ⅱ）. Western blot results showed that compared with the blank group， the levels of phosphorylated extracellular regulated protein kinase （p-ERK）， phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）， phosphorylated c-Jun N-terminal kinase （p-JNK）， and its downstream nuclear transcription factors c-Jun and p-c-Jun protein were significantly increased in M36 group （P<0.05， P<0.01）. The mechanism may be related to the up-regulation of MAPK signaling pathway. ConclusionThe sesquiterpenoid M36 isolated from Myrrha inhibits the proliferation of human hepatoma HepG2 cells and promotes apoptosis and autophagy， which may be related to the activation of the MAPK signaling pathway.
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Objective:To investigate the demographic, epidemiological and clinical characteristics of measles in children, and to explore the risk factors for measles complicated with pneumonia in children.Methods:Children with measles aged≤18 years who were hospitalized in Jinan Infectious Disease Hospital from January 1, 2014 to December 31, 2018 were included. The demographic, epidemiological, clinical and laboratory data of inpatients were collected. The features of patients with pneumonia were analyzed. The risk factors of pneumonia were analyzed by chi-square test and multivariate logistic regression.Results:A total of 1 730 patients were recruited, including 423 patients (24.5%) in 2014, 437 patients (25.3%) in 2015, 856 patients (49.5%) in 2016, 10 patients (0.6%) in 2017 and four patients (0.2%) in 2018. The male to female ratio was 2∶1. The age ranged from four days to 18 years, and 1 572 patients (90.9%) were under three years old. There were 1 381 patients (79.8%) living in rural areas and 83 patients (4.8%) born with low birth weight. Two hundred and twenty-nine patients (13.2%) had a history of respiratory diseases within half a year before measles onset, and 1 489 patients (86.1%) had not been vaccinated before. According to the presence of pneumonia, 1 730 children with measles were divided into pneumonia group ( n=1 295) and non-pneumonia group ( n=435). There were more patients with bucking in pneumonia group than those in non-pneumonia group (56.8%(735/1 295) vs. 40.9%(178/435), χ2=32.770, P<0.01). Zero point seven percent (12/1 730) of children were critically ill, and 0.5%(8/1 730) of children died, all of whom were in pneumonia group. The white blood cell count, the percentage of white blood cell count>10×10 9/L, neutrophilic granulocyte count, the percentage of neutrophilic granulocyte count>7×10 9/L, C reactive protein level, the percentage of C reactive protein level>8.2 mg/L, procalcitonin level, the percentage of procalcitonin>0.5 ng/L, the percentage of hemoglobin level<110 g/L, and the percentage of albumin<35 g/L in pneumonia group were all significantly higher than those in non-pneumonia group ( t=7.153, χ2=47.239, t=8.297, χ2=41.176, Z=-6.769, χ2=40.131, Z=-4.119, χ2=19.284, χ2=7.465, χ2=18.356, respectively, all P<0.01). Multivariate logistic regression analysis showed that male (odds ratio ( OR)=1.316), living in rural areas ( OR=1.521), age<three years old ( OR=2.044), birth weight <2.5 kg ( OR=2.014), respiratory diseases ( OR=1.905), without measles vaccination ( OR=1.591), bucking ( OR=1.550), neutrophilic granulocyte count >7×10 9/L ( OR=3.666), and C reactive protein level >8.2 mg/L ( OR=1.871), procalcitonin>0.5 ng/L ( OR=1.711) were independent risk factors for pneumonia in children with measles. Conclusions:Measles vaccination in children should be farther strengthened. Male, children living in rural areas, low birth weight, history of respiratory diseases before measles and without measles vaccination are prone to be complicated with pneumonia, and special medical attention should be given.
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Gut microbiota is involved in the physiological,biochemical and pathological processes of the body.It could participate in the immunoregulation and the development of enteric nervous system,prevent the invasion of pathogens,maintain normal bowel movement and inhibit the occurrence of tumor.Gut microbiota dysbiosis may also cause many diseases,such as inflammatory bowel disease,obesity and diabetes.A variety of diseases can affect gut microbiota with different ways.In addition,remote organs play a counteractive rule to gut microbiota.Maintaining the balance of the bacteria sets different requirements in different disease processes.This paper reviewed the advanced progression of gut microbiota dysbiosis after cholecystectomy and provided a new therapeutic target for gallbladder diseases.
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Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.