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OBJECTIVE To ex plore the economic value of medication therapeutical management （MTM）service for patients with stable coronary disease. METHODS Totally 140 patients with stable coronary disease were divided into a control group and a intervention group ，70 cases in each group. Patients in control group were received routine medical services ，and patients in intervention group additionally received standardized MTM services on this basis. Medication complication ，satisfaction degree ， safety indexes and efficacy indexes were compared between 2 groups. From the perspective of the whole society ，the economic value of MTM service for patients with stable coronary disease were evaluated by pharmacists using cost minimization analysis. RESULTS A total of 15 patients did not complete the study ，including 5 cases in intervention group and 10 cases in control group ； there was no death endpoint during the follow-up period. MMAS- 8 score，satisfaction score of drug communication dimension and score of overall satisfactionin of intervention group were obviously higher than control group （P＜0.01）. There was no significant difference in blood pressure standard rate ，blood lipid standard rate ，the incidence of adverse drug reaction ，and the incidence of acute coronary events between 2 groups（P＞0.05）. The total cost of intervention group was lower than that of control group （P＜ 0.01）；the results of sensitivity analysis were consistent with those of cost minimization analysis. CONCLUSIONS Pharmacists implement MTM service for patients with stable coronary disease can reduce total cost ，save medical resources and has economic advantages.
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Objective:To investigate the effects of ovarian cancer ascites-derived exosomes on the stem cell properties and invasion ability of ovarian cancer stem-like cell (OCS-LC).Methods:(1) A2780 cells were induced into OCS-LC in serum-free medium, and authenticating their stem-like properties by sphere-forming test, differentiation test and CD 133 marker detection. (2) Exosomes from ascites and A2780 cell were extracted by ultracentrifugation, then authenticating them. The morphology of exosomes was observed by the transmission electron microscope, exosome particle size was measured by nanoparticle tracking analysis (NTA). The expressions of heat shock protein 70 (HSP-70), CD 63 and CD 9 were detected by western blot. (3) The exosomes from ovarian cancer ascites and tumor cell supernate were co-cultured with OCS-LC. The groups were divided into control group, ascites-derived exosomes (ADE) group (ADE+OCS-LC group), and cells-derived exosomes (CDE) group (CDE+OCS-LC group). The sphere-forming ability was evaluated by sphere-forming cycle, maximum sphere diameter and sphere-forming rate of each group; the expression of CD 133 was detected by immunofluorescence staining under microscope; quantitative real-time (qRT)-PCR was used to detected the expression levels of octamer-4 (Oct-4), Nanog mRNA of the signature genes in the stem cells of each group; the metastasis ability of each group was measured by transwell assay. Results:(1) Identification of OCS-LC: sphere-forming experiment showed that the suspension of OCSC single cells was grown in serum-free medium in secondary sphere-forming. Differentiation function experiment showed that OCS-LC were differentiated into adherent A2780 cells by changing the growth mode in serum containing medium. Flow cytometry showed that the proportion of CD 133 positive ( CD133+) cells in OCS-LC group was (18.9±0.9)%, significantly higher than that of control group (0.6±0.5)% ( t=38.570, P<0.01). (2) Under transmission electron microscope, clear lipid bilayer structure was observed in ADE and CDE, and one side presented a concave hemispheric or cup like structure. NTA showed that the diameter of exosomes mainly ranged from 30 to 100 nm, with an average diameter of 67.2 nm. Western blot analysis showed that the specific protein molecules HSP-70, CD 63 and CD 9 were positive. (3) Three groups′ OCS-LC could continue to grow into spheres, and the group of ADE+OCS-LC showed two growth modes, most of the cells continued to grow into spheres, while a small part of cells grew in adherent differentiation. The sphere-forming rate of OCS-LC in the control group, ADE+OCS-LC group, and CDE+OCS-LC group were (1.05±0.20)%, (4.15±0.10)%, and (10.45±0.25)%, the sphere-forming cycle of OCS-LC in the three groups were (15.3±1.5), (10.3±0.6), and (6.7±0.6) days, and the maximum diameters of OCS-LC were (100.3±3.2), (145.2±5.1) and (170.0±2.1) μm, respectively. And the differences were statistically significant (all P<0.05). After co-culture of exosomes with OCS-LC, the sphere-forming ability of cells in the group of CDE+OCS-LC was prior to ADE+OCS-LC group (all P<0.05). Immunofluorescence staining showed that the number of CD 133 green fluorescent chromophore cells in OCS-LC groups [(46.2±2.1)%, (58.4±2.2)%] was significantly higher than that in the control group [(26.6±1.5)%] after the addition of exosomes in co-culture, the positive rate of CD 133 was higher than that in the control group( F=187.588, P<0.05). The qRT-PCR results showed that the expression levels of Oct-4 mRNA in ADE+OCS-LC and CDE+OCS-LC groups were 3.46±0.24, 4.03±0.31, compared with that in control group (1.04±0.12), the differences were statistically significant ( F=134.932, P<0.05). The mRNA expression levels of Nanog were 1.57±0.32, 2.66±0.15, which were significantly higher than that in the control group (1.00±0.07), and the differences were statistically significant ( F=49.329, P<0.05). And the expression of both in CDE+OCS-LC group increased more significantly than ADE+OCS-LC group (all P<0.05). The number of invasive cells in the three groups of OCS-LC were: control group 30±5, ADE+OCS-LC group 102±4, CDE+OCS-LC group 210±7, and there were statistically significant differences among three groups ( F=820.800, P<0.05). Compared with the control group, the number of invaded cells in the co-culture group were significantly increased ( P<0.05), and the CDE+OCS-LC group had the higher cell invasion ability then the ADE+OCS-LC group ( t=23.202, P<0.05). Conclusions:Exosomes derived from ovarian cancer ascites could enhance and maintain the stemness of OSC-LC, and promote the invasion of tumor cells. Moreover, CDE is superior to ADE.
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Objective To discuss the correlation between the effect of vitamin D treatment and vitamin D binding protein gene polymorphism in elderly patients with stable chronic obstructive pulmonary disease(COPD).Methods A total of 800 elderly patients with stable COPD admitted to the four Departments of Respiratory Medicine of four Wenzhou Hospitals were enrolled from September 2016 to November 2018.Because of the drop-out during follow-up,there were final 776 patients in our prospective research.According to the GC gene type,patients were divided into the GC1F-1S(n=214),the GC1F-1S(n=168),GC1F-2 (n=160),GC1S-1S(n=132),GC1F-2(n=82)and GC2-2 groups(n=44),and each group was randomly divided into control group(n=400)receiving the conventional treatment and the treatment group(n=400)taking vitmain D 0.5 μg/d for 3 months as add-on to the conventional treatment.The lung function and the COPD assessment test(CAT)score before and after the therapy were observed and compared in patients with different GC sub-genotypes.Results Among the elderly patients enrolled in our research,GClF-1F genotype was most common,accounting for 26.8％ (214 cases),the followings of next order were GC 1F-1S genotype accounting for 21.0％ (168 cases),and GC2-2 genotype was most rare,accounting for 5.5 ％ (44 cases).The lung function were improved and the CAT score were reduced in all groups after therapy as compared with pre-treatment(P＜0.05).The lung function was better and the CAT score was lower in patients with GC1F-1F,GC1F-1S,GC1F-2,GC1S-1S sub-genotypes(13.77±4.67 vs.15.83±3.73,15.38±4.45 vs.17.86 ± 3.92,17.42 ± 3.19 vs.19.46 ± 4.07) than in the corresponding control subgroups after treatment (P ＜ 0.05).While,there was no significant difference in the lung function and the CAT score between patients with GC1S-2 and GC2-2 genotypes in control versus treatment subgroups(P＞0.05).Conclusions Vitamin D binding protein gene polymorphism is associated with the curative effect of supplement of vitamin D in elderly patients with stable COPD,which has a predictive value for the curative effect of vitamin D supplement.
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Objective@#To discuss the correlation between the effect of vitamin D treatment and vitamin D binding protein gene polymorphism in elderly patients with stable chronic obstructive pulmonary disease(COPD).@*Methods@#A total of 800 elderly patients with stable COPD admitted to the four Departments of Respiratory Medicine of four Wenzhou Hospitals were enrolled from September 2016 to November 2018.Because of the drop-out during follow-up, there were final 776 patients in our prospective research.According to the GC gene type, patients were divided into the GC1F-1S(n=214), the GC1F-1S(n=168), GC1F-2(n=160), GC1S-1S(n=132), GC1F-2(n=82)and GC2-2 groups(n=44), and each group was randomly divided into control group(n=400)receiving the conventional treatment and the treatment group(n=400)taking vitmain D 0.5 μg/d for 3 months as add-on to the conventional treatment.The lung function and the COPD assessment test(CAT)score before and after the therapy were observed and compared in patients with different GC sub-genotypes.@*Results@#Among the elderly patients enrolled in our research, GC1F-1F genotype was most common, accounting for 26.8%(214 cases), the followings of next order were GC 1F-1S genotype accounting for 21.0%(168 cases), and GC2-2 genotype was most rare, accounting for 5.5%(44 cases). The lung function were improved and the CAT score were reduced in all groups after therapy as compared with pre-treatment(P<0.05). The lung function was better and the CAT score was lower in patients with GC1F-1F, GC1F-1S, GC1F-2, GC1S-1S sub-genotypes(13.77±4.67 vs.15.83±3.73, 15.38±4.45 vs.17.86±3.92, 17.42±3.19 vs.19.46±4.07)than in the corresponding control subgroups after treatment(P<0.05). While, there was no significant difference in the lung function and the CAT score between patients with GC1S-2 and GC2-2 genotypes in control versus treatment subgroups(P>0.05).@*Conclusions@#Vitamin D binding protein gene polymorphism is associated with the curative effect of supplement of vitamin D in elderly patients with stable COPD, which has a predictive value for the curative effect of vitamin D supplement.
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Objective To investigate the relationship between the expression of trannsient receptor potential vanilloid 1 (TRPV1) and the severity of airway remodeling in elderly patients with chronic obstructive pulmonary disease(COPD).Methods According to airflow obstruction severity,totally 100 cases of elderly patients with COPD were divided into chronic obstructive pulmonary disease Global Initiative(gold) grade 1 in 23 cases,24 cases of grade GOLD2,GOLD3 27 cases,GOLD4 26 cases,respectively.The TRPV1 concentrations in induced sputum supernatant and serum from each level of elderly patients with COPD as well as in 50 cases of healthy old people were analyzed.Results TRPV1 concentrations in serum and induced sputum in the COPD group was significantly increased compared with the healthy elderly group[(9.94±2.91)μg/L vs.(3.68±0.46)μg/L,(3.29± 1.32)μg/L vs.(0.70 ± 0.30)μg/L] (P ＜ 0.01).The serum and induced sputum TRPV1 concentrations in the mutual pairwise comparison between the elderly COPD patients with all levels had statistical difference (P ＜ 0.01).Conclusion The expression of TRPV1 protein become increased with the severity of airway remodeling in elderly patients with chronic obstructive pulmonary disease.
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Objective To investigate the expression of long intergenic non-protein coding RNA-regulator of reprogramming (Linc-ROR) in high-grade ovarian serous cancer, and explore the relationship between Linc-ROR expression and biological function of high-grade ovarian serous cancer. Methods A total of 34 high-grade ovarian serous cancer tissue samples and 19 normal fallopian tube tissue samples were collected between June 2014 and February 2016. Real-time reverse transcription (RT)-PCR was used to detect the Linc-ROR mRNA expression in different samples. The relationship between Linc-ROR expression level and ovarian cancer International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis was analyzed. Constructed Linc-ROR small interference RNA (siRNA) and pIRES2-EGFP-Linc-ROR plasmid, then Linc-ROR siRNA and pIRES2-EGFP-Linc-ROR plasmid were respectively transfected into SKOV3 cells. Cell proliferation, migration and invasion ability were assessed by cell counting kit-8 (CCK-8), wound healing assay and transwell invasion assay. Results (1) The expression level of Linc-ROR mRNA was significantly higher in high-grade ovarian serous cancer than normal fallopian tube tissues (4.31± 0.38 vs 1.03 ± 0.21; t=25.842, P<0.01). With the progression of FIGO stages, the expression of Linc-ROR was increased (F=95.702, P<0.01), and it was associated with lymph node metastasis (t=7.397, P<0.01). (2) The results of RT-PCR showed that the expression level of linc-ROR in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (0.30 ± 0.11 vs 1.02 ± 0.10; t=15.269, P<0.01). The expression level in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (8.90 ± 0.45 vs 1.03 ± 0.17;t=21.934, P<0.01). The CCK-8 assay showed that when the cells were cultured for 3, 4, 5 and 6 days, the A value in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (P<0.05). And the A value in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (P<0.05). Wound healing assay showed that, after 48 hours incubation, migration rate of cells in Linc-ROR-i group was significantly less than that in the Linc-ROR-NC-i group [(52±4)%vs(67±5)%;t=5.720,P<0.01]. The migration of cells in Linc-ROR-p group was significantly greater than that in the Linc-ROR-NC-p group [(84±4)%vs(66±4)%;t=7.330,P<0.01]. Cell transwell invasion assay showed that, after 48 hours of incubation, the number of invasive cells in Linc-ROR-i group was lower than that in Linc-ROR-NC-i group (74 ± 3 vs 104 ± 3; t=15.810,P<0.01). And the number of invasive cells in Linc-ROR-p group was higher than that in Linc-ROR-NC-p group (217 ± 4 vs 108 ± 5; t=38.060, P<0.01). Conclusion Highly expressed Linc-ROR could enhance the proliferation, migration and invasion ability of high-grade ovarian serous cancer cells, which may be one of the important molecules in the occurrence and development, invasion and metastasis of high-grade ovarian serous cancer.
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Although previous publications suggest the 2009 pandemic influenza A(H1N1)virus was reassorted from swine viruses of North America and Eurasia, the immediate ancestry still remains elusive due to the big evolutionary distance between the 2009 H1N1 virus and the previously isolated strains. Since the unveiling of the2009 H1N1 influenza, great deal of interest has been drawn to influenza, consequently a large number of influenza virus sequences have been deposited into the public sequence databases. Blast analysis demonstrated that the recently submitted 2007 South Dakota avian influenza virus strains and other North American avian strains contained genetic segments very closely related to the 2009 H1N1 virus, which suggests these avian influenza viruses are very close relatives of the 2009 H1N1 virus. Phylogenetic analyses also indicate that the2009 H1N1 viruses are associated with both avian and swine influenza viruses circulating in North America. Since the migrating wild birds are preferable to pigs as the carrier to spread the influenza viruses across vast distances, it is very likely that birds played an important role in the inter-continental evolution of the 2009 H1N1virus. It is essential to understand the evolutionary route of the emerging influenza virus in order to find a way to prevent further emerging cases. This study suggests the close relationship between 2009 pandemic virus and the North America avian viruses and underscores enhanced surveillance of influenza in birds for understanding the evolution of the 2009 pandemic influenza.
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Secretory IgA (SIgA) antibodies in external secretions play an important role in mucosal immune response. Polymeric SIgA was advantageous over monomeric IgA (mIgA) and IgG in several aspects. To express secretory IgA antibody against H5N1 virus, we constructed the secretory component and immunoglobulin J expressing plasmids and co-transfected the plasmids into the Chinese hamster ovary cells (CHO) stably expressing immunoglobulin A. Then we used Zeocin to select the positive clone cells, monoclonal cells stably secreting SIgA was screened through fold dilution method at last. The SIgA antibody secreted from the CHO cells was confirmed by Western blotting, which demonstrated that we had got the complete SIgA molecular. The successful expression of this polymeric anti-H5N1 SIgA in CHO cells will contribute to the production of recombinant SIgA as a preventive agent for infectious disease control.
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Animals , Cricetinae , Antibodies, Viral , Genetics , CHO Cells , Cloning, Molecular , Cricetulus , Genetic Vectors , Immunoglobulin A , Allergy and Immunology , Immunoglobulin A, Secretory , Genetics , Allergy and Immunology , Influenza A Virus, H5N1 Subtype , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , Allergy and ImmunologyABSTRACT
We isolated a novel Enterobacteria phage IME08 from hospital sewage, then confirmed it was a double-stranded DNA phage by digesting its genetic material with DNase I, RNase A and several restriction endonucleases respectively. BLAST results of random fragments generated by a random PCR cloning method revealed that it belonged to T4-like virus. We subsequently determined the host recognizing genes (g37 and g38) sequence with a PCR-based "genome jumping" protocol based on highly conserved region at 5' terminus of g37 from four other T4-like Bacteriophages (T4, JS98, T2 and K3). These molecular biological methods enabled us to readily characterize the bacteriophage and efficiently determine the sequence of the genes of interest based on very limited conserved sequence information.