Eight 1,6-O,O-diacetylbritannilactone (OABL) derivatives (compounds 1-8) were synthesized by esterification or reduction of 1-O-diacetylbritannilactone (ABL) isolated from Inula japonica.All derivatives were evaluated for their anti-inflammation activities through the determination of inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced macrophages.As results,compounds 5-8 (IC50 ＜ 2 μmol/L) exhibited more potent inhibition of NO production activities than the lead compound OABL.

Objective To investigate the expression of Bcl-2 associated athanogene 3 (BAG3) in cervical cancer tissues and cells and its role in epithelial mesenchymal transition (EMT) of cervical cancer.Methods (1) Cervical cancer samples were collected from September 2015 to March 2017 in the Qilu Hospital of Shandong University and Shangdong Provincial Hospital.While,50 normal tissues were collected from August 2015 to March 2017 in the Dezhou Municiple Hospital,which were obtained from patients with uterine mnyoma underwent hysterectomy and patients with cervical biopsy.Reverse transcription (RT)-PCR and western blot were used to detect the expression of BAG3 mRNA and protein,and their clinical significances were analyzed.(2) The expression of BAG3 mRNA and protein was detected using RT-PCR and western blot method in HeLa and SiHa cell lines and normal cervical epithelial cells.The experiment was divided into two groups,BAG3 small interfering RNA transfected group (st-BAG3) and the control group transfected with small interfering RNA (siRNA).Cell counting kit 8 (CCK-8) analysis was used to detect cell proliferation of two groups.Wound-healing and transwell assay were used to detect the migration and invasion ability of HeLa and SiHa cells.The xenograft model of cervical cancer in nude mice was used to observe the effect of BAG3 on tumor xenografts and the tumor-related biomarkers were tested by western blot.Results (1) The expression levels of BAG3 mRNA and protein in cervical carcinoma tissues were 1.20±0.15 and 1.10±0.16,which were significantly higher than that in normal cervical tissue,0.23± 0.04 and 0.29 ± 0.03 (both P＜0.01).The results showed that the expression levels of BAG3 mRNA and protein were significantly correlated with cervical carcinoma staging and lymph node metastasis (P＜0.05).However,its expression was not conrelated with the patient's age,pathological grade,and diameter of tumor (all P＞0.05).(2) Compared with normal cervical epithelial cells,the expression of BAG3 mRNA and protein levels in HeLa and SiHa cells were significantly increased (P＜0.01),the expression levels of BAG3 mRNA and protein in HeLa and SiHa cells transfected with si-BAG3 were significantly lower than that in control group (all P＜0.01).After post-transfected 72 hours,A value of HeLa and SiHa with transfection were significantly lower than those in control group [(0.88±0.08) vs (1.22±0.13),(0.92±0.09) vs (1.35±0.12);both P＜0.01].After post-transfected 24 hours,the migration level of HeLa and SiHa cells with transfection were significantly lower than those in the control group [(20.1±2.1)％ vs (58.6±5.6)％,and (21.1±2.1)％ vs (61.7± 5.4)％;both P＜0.01].The transmembrane cell number in HeLa and SiHa cells with transfection were 76± 11 and 71±8,which were significantly less than those in control group (131± 12 and 129± 14;both P＜0.01).After the inoculation into nude mice,tumor formation time of HeLa and SiHa cells with transfection were (9.5±0.5) and (10.5 ± 1.3) days,respectively,which were significantly longer than those in control group [(4.5±0.5) and (5.2± 1.1) days;both P＜0.05].Compared with those in the control group,the expression level of Slug,N-cadherin and matrix metalloproteinase-2 (MMP-2) protein in HeLa and SiHa cells with transfected in tumor tissues were significantly decreased (all P＜0.01),while the expression level of E-cadberin protein was significantly increased (P＜0.01).Conclusion BAG3 could be involved in the proliferation,migration and invasion of cervical cancer cells by affecting cervical cancer EMT,and BAG3 may be an effective target for the treatment of cervical cancer.

Objective To investigate the risk factors for deep vein thrombosis (DVT) after bone trauma.Methods The study involved 118 patients with traumatic fractures (traumatic fracture group),21 DVT patients diagnosed by color Doppler (DVT group) and 56 healthy patients (control group).Anti-cardiolipin antibody (ACA) was determined by ELISA method.D-dimer and fibrinogen (Fib) were detected by coagulation analyzer and C-reactive protein (CRP) by rate nephelometry.Results Levels of D-dimer,Fib and CRP in traumatic fracture group were significantly increased,but were lower than those in DVT group.ACA positive rate in DVT group presented significant increase and three patients with positive ACA in traumatic fracture group all suffered from DVT.The positive rates of Fib,D-dimer and CRP in lower limb fracture group,multiple fracture group and pelvic fracture group were higher than those in upper limb fracture group (P ＜ 0.05).Levels of Fib and D-dimer showed gradual rise with growth of age,but their levels in DVT group had different degree of reduction after thrombolytic therapy.Conclusions Positive ACA and enhancement of D-dimer,Fib and CRP are risk factors for DVT after bone trauma.Levels of Fib and D-dimer in patients with bone trauma are related with age and therefore risk of posttraumatic DVT increases with age.

Objective To evaluate the effect of As2O3 combined with paclitaxel(PTX)on the treatment of lung cancer.Methods The anti-proliferation efficiency of As2 O3 combined with PTX was evaluated by MTT assay.Tumor spheroids were used to evaluate anti-tumor ability of As2 O3 combined with PTX.Transmission electron microscope (TEM)were used to observe the apoptosis morphous.A549 cell were xenografted in mice to establish the animal model,and the nude mices were devided into four groups,saline group,As2 O3 group,PTX group and As2 O3 +PTX group.The animal model were used to evaluate the effect of anti-tumor.The tumor size of every group were measured.HE was used to observe the apoptosis of cancer cells. Results The cell inhibition rate of A549 cell were(3.35 ±0.21)%,(47.55 ±2.25)%,(64.64 ±3.35)%and(84.58 ±3.76)%after treatment with saline,As2O3,PTX and As2O3combined with PTX after 48h respectively(P<0.01).The early apoptosis rate of cancer cells were 0.26%,9.7%, 17.8% and 42.5% for saline group,As2 O3 group,PTX and As2 O3 +PTX group respectively(P<0.01 ).The final tumor spheroid volumes in saline group increased 1.36 times after 7 days.The final tumor spheroid volumes reduced to(77.35 ±2.31)%,(61.68 ±2.44)% and(44.85 ±3.34)% in As2O3,PTX and As2O3 combined with PTX group respectively(P<0.01).The inhibition of lung cancer in vitro demonstrated the inhibition rate of tumor growth compared with saline group were(22.4 ±4.5)%,(39.5 ±6.2)% and(69.5 ±7.3)% for As2O3,PTX and As2O3 +PTX,respectively(P<0.01 ).Conclusion As2 O3 combined with PTX can effectively inhibit the proliferation of A549 cells and ectopic tumor growth in nude mice and it may be a potentially effective treatment for lung cancer.

The aim of this study is to identify the express specificity of bone morphogenetic protein 15 (Bmp15) in porcine. The pBMP15-EGFP reporter vector was constructed from the 2.2 kb fragment of porcine bmp15 promoter to trace the differentiation process of stem cells into oocyte-like cells. We used porcine ovary and Chinese Hamster Ovary cell line (CHO), mouse myoblast cell line (C2C12) and porcine amniotic fluid stem cell (pAFSC) to investigate the expression and regulation of this gene via RT-PCR, immunofluorescence, cell transfection, and microinjection methods. We also used single layer cell differentiation to detect the application potential of bmp15. The results show that bmp15 gene was specifically expressed in the porcine ovary and CHO rather than in C2C12 and pAFSC. In addition, the characteristic of tissue-specific of Bmp15 was detected on CHO instead of other cell lines by transient transfection. We also detected the expression of Bmp15 in oocyte at different development stages by immunofluorescence of fixed paraffin-embedded ovary sections. Furthermore, microinjection results show that bmp15 expressed in oocytes at 18 h of maturation in vitro, and continued up to 4-cell stage embryos. Most importantly, we found that the expression of Bmp15 started at day 12 after inducing pAFSC into oocyte-like cells by transfection; green fluorescent was visible in round cell masses. It indicated that bmp15 has the expression specificity and the pBMP15-EGFP reporter vector can be used to trace Bmp15 action in the differentiation of stem cells into germ cells.
Animals ; Bone Morphogenetic Protein 15 ; genetics ; CHO Cells ; Cell Differentiation ; Cricetinae ; Cricetulus ; Female ; Genes, Reporter ; Genetic Vectors ; Mice ; Microinjections ; Myoblasts ; cytology ; Oocytes ; metabolism ; Ovary ; metabolism ; Stem Cells ; cytology ; Swine

Hemagglutinin (HA) contains a head domain with a high degree of variability and a relatively conserved stem region. HA is the major viral antigen on the surface of the influenza virus. To define the biologic activities of chimeric HA bearing different head domains and stem regions or their potential use, a HA chimeric gene containing the head domain of the H7 subtype virus and stem region of the H3 subtype virus was modified and expressed using a baculovirus expression vector. Then, the secreted protein was purified and its biologic activities characterized. Approximately 1.4 mg/mL cH7/3 HA could be obtained, and its molecular weight was ≈ 70 kD. The trimer form of cH7/3 protein had hemagglutination activity and could be recognized by specific antibodies. The method described here can be used for further studies on the screening of HA stem-reactive antibodies or the development of vaccines with conserved epitopes.
Antibodies, Viral ; immunology ; Baculoviridae ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza Vaccines ; genetics ; immunology ; Influenza, Human ; prevention & control ; virology

Objeetive:To observe the effect of acupuncture-moxibustion on TNF-a,sTNFR-Ⅰand sTNFR-Ⅱ in rats with Crohn's disease(CD).Method:The models of CD rats induced by TNBS were randomized into model group.herbal cake-partitioned moxibustion group and electroacupuncture group as well as a normal control group.The histopathological changes of colon mucus membrane were observed with HE staining and the contents Of TNF-a.sTNFR-Ⅰand sTNFR-Ⅱ were detected with ELISA method.Results:When compared with the normal group,the TNF-a level in CD rats was substantially elevated and the sTNFR-Ⅰ and sTNFR-Ⅱ showed no marked changes.After herbal cake-partitioned moxibustion and electroacupuncture treatment,the TNF-a level in CD rats was substantially reduced and sTNFRL-Ⅰ and sTNFR-Ⅱ showed no marked changes.The inflammatory lesions and abnormal structures of CD rats were significantly improved after herbal cake-partitioned moxibustion and electric stimulation.Conclusion:Herbal cake-partitioned moxibustion and electroacupuncture Can both remarkably reduce the TNF-a level in blood serum.and this might be one ofthe action mechanisms in the treatment of CD.

Objective To investigate correlation between aspirin resistance(AR) and inflammatory factors. Methods One hundred and ten patients with coronary heart disease took aspirin 0.1 mg/d for 14 days.It was detected platelet aggregation function induced with adenosine disphosphate (ADP) and arachidonic acid (AA), and investigated correlation between AR and inflammatory factors. Interleukin-1? (IL-1?),interleukin-6 (IL-6) and high sensitive C-reaction protein (hs-CRP) levels. Results IL-6 level of patients with AR was significantly higher than that of aspirin sensitive (AS) patients. The other two index were not different between the two groups. Conclusion IL-6 levels could be used as predictor.

Objective To investigate the characteristics of gut microbiota in SD rat model of diabetes mellitus induced by streptozotocin (STZ). Methods Twenty-five male SD rats were randomly divided into control (C) (n＝10) and diabetes (M) (n＝15) groups. Rats in the group M received intravenous injection of streptozotocin (30 mg/kg) once per day for 5 consecutive days. Fecal samples were collected and examined for the V3 region of the 16S rDNA gene by Illumina Miseq high-throughput sequencing. The abundance and composition of gut microbiota were analyzed by cluster analysis. Results DNA sequence analysis was successfully performed. The Chao 1 index was lower in the group M than group C (P＜ 0. 05). The Shannon index was lower and the Simpson index was higher in the group M than group C (P＜0. 05). At phylum level, the relative abundance of Proteobacteria, Cyanobacteria, Tenericutes, TM7, and Actinobacteria was lower in the group M than group C (P＜ 0. 05). At genus level, 4 weeks after injection,the abundance of Lactobacillus was lower and that of Bacteroidetes was higher in the group M than group C ( P＜ 0. 05). 12 weeks after injection, the relative abundance of Lactobacillus,Bacteroides and Ruminococcus was higher and that of Bifidobacterium was lower in the group M than group C ( P＜ 0. 05). Conclusions This STZ-induced diabetic SD rat model has a low abundance and diversity of gut microbiota. Quantitative analysis of gut microbiota composition in this animal model provides a basic data for the study of relationship between diabetes mellitus and gut microbiota.

Objective:To evaluate the role of interleukin-6 (IL-6) and CD4 + T-lymphocytopenia in assessing the severity and prognosis of coronavirus disease 2019 (COVID-19). Methods:A prospective observational study was conducted. Forty-five patients with COVID-19 admitted to Henan Provincial People's Hospital from January 13 to March 13, 2020 were enrolled and divided into normal group (13 cases), severe group (20 cases), critically severe group (12 cases) according to the severity of the disease. A total of 15 healthy subjects receiving physical examinations during the same period were collected as the healthy control group. Clinical data were collected to compare the clinical characteristics, general test results, IL-6 and CD4 + T-lymphocytopenia levels of patients in different disease severity groups and healthy control group. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of each indicator for the severity of COVID-19. Multivariate Cox regression analysis was used to analyze the risk factors affecting the prognosis of COVID-19 patients, and Kaplan-Meier survival curve analysis was performed. Results:The age of the critically severe group was significantly higher than that of the severe and normal groups (years old: 66.91±17.01 vs. 59.35±18.07, 40.23±12.61, both P < 0.05), and the negative conversion time of the 2019 novel coronavirus (2019-nCoV) was significantly longer than that of the severe and normal groups (days: 19.00±10.66 vs. 18.00±7.18, 9.31±3.49, both P < 0.05). With the increase of the severity of disease, white blood cell count (WBC), C-reactive protein (CRP), calcitonin (PCT), total bilirubin (TBil), troponin I (TnI), IL-6, D-dimer and other indicators were significantly increased, while lymphocyte count (LYM), platelet count (PLT), CD4 +, CD8 +, oxygenation index (PaO 2/FiO 2) were significantly decreased (all P < 0.01). ROC curve showed that PaO 2/FiO 2, IL-6 and CD4 + had certain predictive value for disease severity of COVID-19, the area under the ROC curve (AUC) of them were 0.903, 0.871, 0.689, and the 95% confidence interval (95% CI) were 0.806-0.949, 0.769-0.974, 0.542-0.853; the best cut-off values were 196.00 mmHg (1 mmHg = 0.133 kPa), 6.02 ng/L, 355 cells/μL, respectively; the sensitivity were 73.3%, 99.3%, 73.3%, and the specificity were 96.6%, 62.1%, 65.5%, respectively. Multivariate Cox regression analysis showed that age, PaO 2/FiO 2, high IL-6 and low CD4 + (IL-6≥6.02 ng/L and CD4 + < 355 cells/μL) were independent risk factors affecting the prognosis of COVID-19 [hazard ratio ( HR) was 1.077, 0.053 and 3.490, respectively, all P < 0.05]. Kaplan-Meier survival analysis showed that when both high IL-6 and low CD4 + (IL-6≥6.02 ng/L and CD4 + < 355 cells/μL) were present, the mean time of adverse prognosis was (20.53±5.71) days; when increased IL-6 and decreased CD4 + were inconsistent, the mean time of adverse prognosis was (53.21±3.16) days. Conclusions:The levels of IL-6 and CD4 + T-lymphocytopenia are closely related to the severity of COVID-19 disease. When IL-6 ≥ 6.02 ng/L and CD4 + < 355 cells/μL occur simultaneously, the prognosis is poor.