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Objective To explore the improvement of pulmonary function by tiotropium bromide for chronic obstructive pulmonary disease (COPD)patients with stable phase,and to evaluate its effect on inflammatory factors, thus to provide the basis for clinical treatment.Methods 124 COPD patients with stable phase were selected in the hospital from November 2014 to August 2015.According to the admission number with the single and double number, they were randomly divided into the control group and the study group,62 patients in each group.The control group was given conventional treatment,and the study group was given tiotropium on the basis of conventional therapy. Before and after treatment,the forced vital capacity (FVC),forced expiratory volume in one second (FEV1 )and FEV1 /FVC index were assessed in two groups,the serum levels of high -sensitivity C -reactive protein (hs -CRP) and tumor necrosis factor -α(TNF -α)and other inflammatory factors were measured before and after treatment. Results FEV,FEV1 and FEV1 /FVC(%)of the study group after treatment were (1.79 ±0.36)L,(1.24 ±0.29)L and (67.74 ±1.78)% respectively,which were higher than those of the control group [(1.68 ±0.31)L,(1.03 ± 0.25)L and (62.91 ±1.84)%],the differences were statistically significant (t =9.64,9.28,9.60,all P <0.05). The serum levels of hs -CRP and TNF -αof the study group after treatment were (6.10 ±0.38)mg/L and (6.12 ±0.42)ng/ml,which were lower than those of the control group [(1.54 ±0.09)g/L,(8.29 ±0.31 )ng/mL],the differences were statistically significant (t =12.51,9.24,all P <0.05 ).Conclusion Application of tiotropium bromide for the stable COPD patients can significantly improve the patients'lung function,lower levels of inflammatory factors.And it has positive clinical significance in stable condition,slow disease progression.
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This study was aimed to explore the impact of activated carbon adsorption modified by different concentrations of nitric acid and ammonia,in order to examine the impact on adsorption of mannotriose by modified activated carbon.The activated carbon was processed by different concentrations of nitric acid and ammonia.And then,the adsorption capacity of benzene,iodine,methylene blue and the purification effects on mannotriose were measured.The results showed that the active carbon modified by nitric acid and ammonia had some changes of adsorptions for methylene blue,iodine and benzene.The purification effect of mannotriose with nitric acid-modified activated carbon was declined.The purification effect of mannotriose with ammonia-modified activated carbon was increased.It was concluded that the pore structure of activated carbon had been changed by nitric acid and ammonia.The adsorption capacity of nitric acid modified active carbon to mannotriose was declined.However,the adsorption capacity of ammonia modified active carbon to mannotriose was increased.It showed that ammonia modified active carbon was suitable for the purification of mannotriose.And the adsorption capacity of iodine reflected the adsorption capacity of mannotriose by active carbon.
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This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.
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<p><b>OBJECTIVE</b>To assess the effect of platelet rich plasma (PRP) on proliferation and differentiation of human MG63 osteoblast-like cells and the biological function of PRP in vitro.</p><p><b>METHODS</b>PRP was obtained from venous blood of a health volunteer by two step centrifugation. CaCl2 and thrombin were used to activate PRP. The differentiation of MG63 cells, which were exposed to various concentrations of PRP (0, 1%, 2%, 3%) was detected by alkaline phosphatase (ALP) activity. Propidium iodide (PI) fluorescent coloration staining was used to observe the morphology of cells. Immunocytochemistry was used to evaluate the expression level of transforming growth factor-beta (TGF-beta) in MG63 cells in different concentration of PRP. The cells adhered to calcium phosphate material was observed by scanning electron microscopy (SEM). The proliferation was evaluated by cell counting kit-8 (CCK-8) proliferation assay. The cell cycle assay was performed by low cytometry (FCM) to detect the effect of PRP on MG63 cells in different time points. The mRNA level of Col-I in MG63 cells cultured under different concentration PRP was checked by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>ALP activity experiment demonstrated that the maximum effect was got in 3% PRP group. PRP had a positive effect on the proliferation of MG63 cells but cells also presented disengage phenomena from the glass slides. The PI staining showed that PRP improved fluorescent intensity of cell nucleus. Immunocytochemistry showed that TGF-beta expression level was significantly enhanced on 3% PRP group (P<0.05). SEM showed that cells grew well on material in PRP group. The results of CCK-8 showed that the mean absorbency number A(450 nm) of 4.8% PRP was significantly higher than that of control group (P<0.05). FCM showed that S period cells percentage of PRP group was higher than that of control group in the 2nd day (P<0.05); G0/G1 period cells percentage of PRP group was significant increased than that of control group in the 10th day (P<0.05); G2/M period cells percentage of PRP group was higher than that of control group except the 6th day. PRP promoted the expression of Col- I in MG63 cells by RT-PCR.</p><p><b>CONCLUSION</b>These data suggest that PRP has a positive influence on MG63 proliferation, transference and the expression of relative protein and gene in an appropriated concentration. The findings of this study also demonstrated that PRP may play a beneficial role of unifying and modulating the biological behavior of MG63 cells.</p>
Subject(s)
Humans , Alkaline Phosphatase , Cell Differentiation , In Vitro Techniques , Osteoblasts , Platelet-Rich Plasma , Transforming Growth Factor betaABSTRACT
This paper is aimed to isolate and to cultivate human adipose-derived stem cells (hADSCs) from the adipose tissue by a combination of collagenase digestion, adherence to flasks and monoclonal cultural method so as to observe the morphological characteristics of the hADSCs. The immunophenotypes of hADSCs were detected by flow cytometry techniques. The general morphological characteristics of hADSCs were observed by cytochemical and immunofluorescent techniques. The ultrastructure of hADSCs was observed by transmission electron microscopy (TEM). The experimental results showed that hADSCs had unique immunophenotypes and they were positive for CD29, CD44, CD90, CD105 and CD166, but negative for CD31, CD45 and HLA-DR. Cytochemistry showed that cytoplasm of hADSCs was stained with light blue by hematoxylin-eosin, negative for Oil red O and AKP, and positive for immunofluorescence CD29 and CD166. There were abundant organella and microvilli in the ultrastructure of hADSCs. The results validate that they will offer a morphological foundation for application of the hADSCs.
Subject(s)
Humans , Adipose Tissue , Cell Biology , Cell Culture Techniques , Methods , Cells, Cultured , Microscopy, Electron, Transmission , Stem Cells , Cell Biology , Tissue Engineering , MethodsABSTRACT
We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.
Subject(s)
Humans , AIDS Vaccines , Allergy and Immunology , Escherichia coli , Genetics , Metabolism , HIV-1 , Genetics , Mutation , Peptide Fragments , Genetics , Allergy and Immunology , Peptide Library , Recombinant Proteins , Genetics , Allergy and Immunology , tat Gene Products, Human Immunodeficiency Virus , Genetics , Allergy and ImmunologyABSTRACT
BACKGROUND:Up to date,there is not an acceptable method for isolating,culture and amplifying human adipose-derived mesenchymal stem cells (hADSCs).OBJECTIVE:To explore the most effective way to obtain highly homogenous and undifferentiated hADSCs.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the Key Laboratory of Pathobiology,Ministry of Education,Jilin University from June to December 2008.MATERIALS:Human abdominal adipose tissue resected in the surgery was supplied by the Affiliated Hospital of Jilin University,The informed consent was obtained from patients.METHODS:Human adipose tissue was removed connective tissue and blood vessel,followed by incubation in 0.1% type I collagenase for 60 minutes at 37℃,filtrated then centrifuged.Consequently,the subnatant precipitation was cultured with LG-DMEM containing 10% fetal bovine serum,incubated at 6-well plate with density of 1×10~9/L,and placed in incubator of 5% CO_2 at 37 ℃.The cultured cells were passaged when the cells reached 80%-90% confiuency,and the 3~(rd) passage of cells were induced to osteogenic and adipogenic differentiation.MAIN OUTCOME MEASURES:Morphological characteristics of hADSCs were observed by laser scanning confocal microscope.Immunophenotypes,cell cycle and growth curve of hADSCs were detected by flow cytometry and immunofiuorescent techniques.In addition,the multiple differentiation potential of hADSCs was detected.RESULTS:hADSCs presented fibroblast-like morphological feature with a flocked array.The 3~(rd) passage of hADSCs had unique immunophenotypes and they were positive for CD73,CD44,CD166,CD105 and CD29,but negative for CD31,CD34,CD45 and HLA-DR.Cell cycle result showed that they had the typical growth characteristics of stem calls,namely,83.81% cells stayed at G_0/G_1 stage,only 16.19% cells were stayed at S+G_2/M stage;The latent phase of the primary culture cells was 2 days prior to and after incubation,followed by 3-6 days of logarithmical proliferation,reached a peak at day 6,and entering the growth platform phase with lower growth speed.The alkaline phosphatase was positive expressed at week 2 of osteogenic induction.And the positive expression of oil-O red staining could be seen at day 3 of adipogenic induction.CONCLUSION:Cells contamination can be reduced by removing connective tissue and blood vessel of adipose tissue,and 0.1% type Ⅰ collagenase for 60 minutes can effective separated stroma cell to matrix fiber,furthermore,ensure a sufficient contact between collagenase and tissures,which provide an supportive for harvesting highly homogenous hADSCs.
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OBJECTIVE: To intensify ward management on high-alert medications ensure clinical medication safety.METHODS: The main problems encountered in ward management on high-alert medications were analyzed;the application of continuous quality improvement(CQI) in the management on high-alert medications was introduced and the outcome was evaluated.RESULTS: The continuous quality improvement has greatly enhanced the management quality on high-alert medications and reduced the number of problems from previous 158 times before practice to 2 times after the practice of the continuous quality improvement.CONCLUSION: CQI is a continuous activity and it is conducive to the improvement of management quality on high-alert medications.