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BACKGROUND:Human placental mesenchymal stem cells have been shown to be effective in inhibiting the development of pulmonary fibrosis,but the underlying mechanisms remain unclear. OBJECTIVE:To investigate the therapeutic effect and related mechanism of human placental mesenchymal stem cells on silica-induced pulmonary fibrosis in human embryonic lung fibroblasts(MRC-5). METHODS:CCK-8 assay was used to detect the effects of different mass concentrations of silica on the proliferation of MRC-5 at different time points.Immunofluorescence staining was used to screen out the best stimulating mass concentration and time of silica for subsequent experiments.MRC-5 cells were divided into blank group,silica group,and silica + human placental mesenchymal stem cell group.In the blank group,cells were not treated.In the silica group,MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours.In the silica + human placental mesenchymal stem cell group,MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours and then co-cultured with human placental mesenchymal stem cells for 24 hours.Immunofluorescence staining was used to detect the expression of α-smooth muscle actin and collagen type I in cells of each group.Western blot assay was used to detect the expressions of pulmonary fibrosis-related proteins and TGF-β1/Smad 3 signaling pathway-related proteins in cells of each group. RESULTS AND CONCLUSION:(1)CCK-8 assay results suggested that 100 μg/mL silica was the best mass concentration and time to stimulate MRC-5 cells for 48 hours.(2)Immunofluorescence staining results showed that the expression of α-smooth muscle actin and collagen type I in the silica + human placental mesenchymal stem cell group was significantly lower than that in the silica group.(3)Western blot assay results showed that compared with the silica group,the protein expression levels of α-smooth muscle actin,collagen type I,N-cadherin,fibronectin,transforming growth factor-β1,p-Smad3,and Smad3 in the silica + human placental mesenchymal stem cell group were decreased,and the expression of E-cadherin was increased.The difference was statistically significant(P<0.05).(4)The results showed that human placental mesenchymal stem cells had a significant therapeutic effect on silica-induced pulmonary fibrosis.Human placental mesenchymal stem cells can inhibit the development of pulmonary fibrosis by regulating transforming growth factor-β1/Smad3 signaling pathway.
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@#To investigate whether rare ginsenosides could alleviate idiopathic pulmonary fibrosis (IPF), C57BL/6 mice were randomly divided into control group, bleomycin (BLM)-induced IPF group, rare ginsenoside Rk1 group, rare ginsenoside Rk3 group, rare ginsenoside Rh4 group and rare ginsenoside Rg5 group.All mice except those in the control group were given bleomycin injection.The IPF model was established by BLM for 28 days.The treatment group was given ginsenoside intragastrically at the same time.After the experiment, the lung tissues of mice were collected and the pathological changes of the mice lungs were observed.The content of hydroxyproline (HYP) in mouse lung tissue was measured.The expression of IPF-related genes in mouse lung tissues was detected.In in vitro experiments, Medical Research Council cell strain-5 (MRC-5) was used to induce IPF cell model using transforming growth factor-β1 (10 ng/mL).The effects of four saponins on the expression of IPF-related genes were analyzed by MTT assay, HYP content determination and RT-qPCR.All four rare ginsenosides could effectively alleviate the pathological process such as alveolar structure destruction caused by IPF, reduce the content of HYP, and down-regulate the expression of IPF-related genes, indicating that rare ginsenosides can effectively alleviate IPF.
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@#Abstract：Objective To improve the replication level of varicella⁃zoster virus（VZV）in human diploid cell line MRC⁃5 and increase the yield of VZV vaccine by reducing the expression of interferon（IFN）related genes via optimizing the cell line MRC⁃5. Methods Interferon receptor 1（IFNAR1）silenced MRC⁃5 cell line（MRC⁃5IFNAR1⁃）was constructed by CRISPR／Cas9 gene editing technology，which was determined for the relative expression of IFNAR1 mRNA，and for those of mRNA of IFN related genes IFNβ and OAS1 after VZV infection by qRT⁃PCR to evaluate the effect of gene silencing. Gene mutation sequences were further identified by sequencing of the silenced sites. The replication of VZV in MRC⁃5 and MRC⁃5IFNAR1⁃ cell lines was compared 168 h after VZV infection by using qRT⁃PCR and plaque formation unit（PFU）assay， to evaluate the effect of MRC⁃5IFNAR1⁃cell line on VZV replication. Results The growth status of MRC⁃5IFNAR1⁃ cell line wasconsistent with that of MRC ⁃ 5 cells，and the relative expression of IFNAR1 mRNA decreased by 73%；The relative expressions of IFNβ and OAS1 mRNA in MRC⁃5IFNAR1⁃ cell line were 61% and 90% lower than those in MRC⁃ 5 cells respectively after VZV infection；In addition，168 h after VZV infection，the level of DNA replication and the titer of VZV increased by 5. 7 folds and 4 folds respectively. Conclusion The successful establishment of MRC⁃5IFNAR1⁃ cell line may be a potential scheme to increase the yield of vaccines based on human diploid cells，and provided a reference for expanding production of VZV vaccine.
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@#Abstract：Objective To investigate the adaptability and genetic stability of hepatitis A virus（HAV）SYX1 strain in human diploid cell MRC⁃5. Methods HAV SYX1 strain isolated from feces of patients with hepatitis A was continuously propagated in MRC⁃5 cells for 28 passages，of which the 1st ~ 26th passages were determined for antigen contents and virus titers，the 6th passage was observed for the morphology under microscope and detected for physicochemical properties，and the 13th ~ 15th passages were studied for virus proliferation dynamics to determine the peak yield of virus proliferation. Genomic RNA was extracted from the 8th，12th，18th，20th，22nd，25th，26th and 28th passages and sequenced to analyze the genetic stability. The main seed batch and working seed batch of HAV SYX1 strain were established and verified according to the requirements of Chinese Pharmacopoeia（VolumeⅢ，2020 edition）. Results The antigen content of HAV SYX1 was stable at 160 ~ 320 EU／mL and the titer was maintained at 7. 3 ~ 8. 3 lgCCID50／mL after the 8th passages in MRC 5 cells；Virus particles showed two types：hollow and solid，with a diameter of 27 ~ 32 nm，spherical，without envelope and protrusions on the surface，which tolerated low pH value and ether. The peak period of virus proliferation was 10 d with an antigen content of more than 160 EU／mL and a virus titer of more than 7. 0 lgCCID50／mL. HAV SYX1 was a subtype of HAV IB，and no mutation in the coding region of all structural proteins during passage was observed. The verification results of main seed batch and working seed batch of HAV all met the relevant requirements. Conclusion HAV SYX1 strain showed good adapt⁃ ability and genetic stability in MRC⁃5，which might be used for the development and production of inactivated hepatitis A vaccine.
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To evaluate the immunogenicity of BCG-CpG-DNA-adjuvanted human rabies vaccine(MRC-5 cell)in cynomol-gus monkeys,we randomly divided monkeys into a control group,three-dose group,and four-dose group.The three-dose and four-dose groups were vaccinated with BCG-CpG-DNA-adjuvanted human rabies vaccine for human use at 0,7,and 21,or at 0,3,7,and 14 days,respectively.The control group was vaccinated with a commercially available domestic vaccine with a 2-1-1 protocol.Serum samples were collected from all monkeys before(0 day)and after immunization,on days 7,14,28,35,49,and 63.RFFIT and ELISPOT were used to detect anti-rabies virus neutralizing antibodies and cytokines(IFN-γ and IL-2).The anti-rabies virus neutralizing antibodies in all groups were negative before immunization.The three-dose and control groups showed an increasing trend from 7 to 28 days after primary immunization,and the highest level was reached on the 28th day;in contrast,the four-dose group showed a peak on the 14th day.On the 28th day,the neutralizing antibody levels in the control and three-dose groups were significantly higher than those in the four-dose group.On the 35th day after the initial immuniza-tion,the neutralizing antibodies in the three-dose group was significantly higher than those in the four-dose group.The levels of IL-2 and IFN-γ in peripheral blood lymphocytes in the three-dose and four-dose groups tended to increase from 7 to 14 days and peaked on the 14th day;the level of IFN-γ in the three-dose group was significantly higher than that in the control group on the 14th day.Otherwise,no significant differences in the levels of neutrali-zing antibodies and cytokines were observed among all groups at all other time points.Thus,BCG-CpG-DNA adjuvanted human rabies vaccine(MRC-5 cell)has excellent immunogenicity and application value for optimizing immunization procedures.
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Aim To explore the role of transcription factor F0XM1 in collagen synthesis in MRC-5 cells in-duced by high glucose.Methods I ▪ The optimal time and concentration of the hyperglycemia model in MRC-5 cells were explored by CCK8: the time gradi¬ents: 6,12,24,48,72 h; the concentration gradients: 5.5,15 ,30,45 mmol • L 1 , and 30 mmol • L 1 man- nitol was used to be the hypertonic control group.(2) Hie effect of collagen synthesis in MRC-5 cells induced by high glucose was detected: the cells were divided into normal control group, hypertonic control group (30 mmol • L 1 mannitol) and high glucose (30 mmol • L 1 ) group.WB and qPCR assays were used to de¬tect the expression of conllagen synthesis factors ( Fn, COL 1, COL m, a-SMA, MMP9, TIMP1 ) and TGF-p signaling pathway factors (TGF-pi , p-Smad2/ Smad2).(3 The role of FOXMl in promoting collagen synthesis by high glucose was investigated: the cells were divided into normal control group, hypertonic control group (30 mmol • L 1 mannitol) , high glucose ( 30 mmol • L 1 ) group and high glucose (30 mmol • L 1 ) + thiostrepton ( 1 (xmol) group, and the expres¬sions of FOXM1 , collagen synthesis factors were detec¬ted by WB and qPCR assays.Results Mannitol had no significant effect on proliferation of MRC-5 cells, hut their proliferation activity was significantly lower than that of control group when MRC-5 cells were trea¬ted with 30 mmol • L 1 high glucose for 24 h; the ex¬pressions of COL 1 , COL IH , F0XM1 and other fac¬tors were promoted when MRC5 cells were treated with high glucose; the expression of F0XM1 was signifi¬cantly inhibited after the addition of thiostrepton, and the expressions of collagen synthesis factors also de-creased compared with high glucose group, and the a- bove differences were all statistically significant (P < 0.05).Conclusion FOXM1 is a factor related to the increase of collagen synthesis in MRC-5 cells induced by high glucose.
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Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.
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OBJECTIVE: To investigate the differentially expressed microRNAs(miRNAs) in human embryonic lung fibroblast MRC-5 cells stimulated by transforming growth factor-β1(TGF-β1) using microarray chip, and screen for key genes and signaling pathways of fibroblast trans-differentiation. METHODS: The miRNA expression gene chip dataset GSE43992 on TGF-β1 stimulated MRC-5 cells were downloaded from high-throughput Gene Expression Omnibus(GEO) database of National Center for Biotechnology Information of the United States. The R language Limma package was used to screen the differentially expressed miRNAs. Corresponding target genes were predicted by miRWalk database performed by Gene Ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analysis. The protein-protein interaction(PPI) network was constructed by the search tool for the Retrieval of Interacting Genes database. RESULTS: A total of five differentially expressed miRNAs were identified, including four up-regulated miRNAs and one down-regulated miRNA; and 42 corresponding differentially expressed target genes were predicted. GO analysis indicated that the target genes were significantly enriched in collagen catabolic process, extracellular matrix organization, membrane organization, collagen fibril organization, and cellular response to amino acid stimulus. The results of KEGG pathway analysis showed that the signaling pathways corresponding to miRNAs and target genes were mainly concentrated in 18 signaling pathways, that were mainly related to the age-ethnic signaling pathways and protein digestion and absorption miRNAs in tumors and diabetic complications. The core genes transfected into the myofibroblasts by the three fibroblasts screened by the PPI network were threonine kinase 1, estrogen receptor 1 and β-catenin. CONCLUSION: Five differentially expressed miRNAs, 42 target genes, 18 signaling pathways, and 3 core genes related to TGF-β1-induced MRC-5 cell trans-differentiation were screened. It can provide new reference for the treatment and research of many diseases including pneumoconiosis and pulmonary fibrosis.
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Objective @#To observe the changes of soft tissue in patients with Angle class Ⅱ division Ⅰ malocclusion during mixed dentition treated with MRC functional appliance.@* Methods @# Twenty patients with Class Ⅱ division Ⅰ malocclusion of Angle were treated with functional MRC. The facial features before and after treatment were measured by software and the results were analyzed statistically. @*Results@#The patients′soft tissue profiles were improved significantly before and after treatment, The OE-Prn-Pos angle, OE-N′-B′ angle, OE-N′-Pos angle, OE-Prn-N′angle, Cm-Sn-UL angle, and N′-Sn-Pos angle increased significantly (P < 0.05). The OE-Sn-UL angle, and Sn-N′-B′ angle decreased significantly (P < 0.05); the distance between the lateral soft tissue line and the middle Sn-H line, UL-E line and LL-E line were significantly different (P < 0.05). The distances were all reduced, and the difference was statistically significant (P < 0.05).@* Conclusion @#The application of an MRC functional appliance can improve the relationship among nasolabial soft tissue, upper and lower lip soft tissue, and chin-lip soft tissue, thus improving the protrusion profile of patients.
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Objective To investigate the effect of simvastatin on oxidative stress and inflammatory reac-tion in patients with stable moderate to severe chronic obstructive pulmonary disease and its mechanism. Meth-ods Sixty patients diagnosed with chronic obstructive pulmonary disease were randomly divided into the simvas-tatin group and the placebo group.The simvastatin group was treated with simvastatin in 40 mg/d for 12 weeks,and the placebo group with placebo.The general clinical features,the concentration of inflammatory factors,pulmonary function,6-minute walk test and MRC score were compared between the two groups.Results There was no signif-icant difference between these two groups in basic features. There was a decrease of IL-6,TNF-a and Hs-CRP in concentration in the simvastatin group after treatment. which was significantly lower than that of the placebo group after treatment.The 6-minute walk test in the simvastatin group was much better than that in the placebo group(P=0.00).MRC score was improved compared with therapy before(P=0.02).There was no significantly difference in 6-minute walk test and MRC score before and after treatment in the placebo group(P=0.81). The PaO2 was im-proved after treatment in the simvastatin group compared with therapy before and that in the placebo group after therapy(P<0.05)respectively.There was no significantly difference in FEV1and FVC between these two groups. Conclusion Simvastatin can decrease the concentration of inflammatory factors in stable moderate to severe chron-ic obstructive pulmonary disease,and improve the pulmonary function.
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Aim To examine the role and uderlying mechanisms of Lin28 /let-7d axis in the proliferation of lung fibrobalsts and fibroblasts-into-myfibroblasts tran-sition,and provide novel strategy for the treatment of idiopathic pulmonary fibrosis (IPF).Methods We induced experimental lung fibrosis in mice by intratra-cheally injection of bleomycin (BLM).Ang Ⅱ and TGF-β1 were used to induce fibrogenesis in cultured MRC-5 cells;qRT-PCR and Western blot were applied to determine the changes of Lin28B,collagen 1 α1 and collagen 3α1 ;MTT assay,Edu satining and immun-ofluoresence were used to examine the cell viability, proliferation and fibroblasts-into-myofibroblasts transi-tion in MRC-5 cells.Results Lin28B was increased in the lung of mice with experimental lung fibrosis and in MRC-5 cells treated with AngⅡ or TGF-β1 .Moreo-ver,Lin28B enhanced collagen deposition via inhibi-ting expression of let-7d,which maybe contribute to the progression of IPF.In addition,further studies showed that Lin28B promoted proliferation and fibro-blasts-into-myofibroblasts in MRC-5 cells.Conclusion Lin28B /let-7d axis contributes to fibrogenesis via promotes fibroblasts-into-myofibroblasts transition, which may provide novel approaches for lung fibrosis treatment.
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Objective To investigate the effect of melatonin on oxidative stress and inflammatory reaction in patients with moderate to severe chronic obstructive pulmonary disease and to explore its mechanisms. Methods 42 patients with moderate to severity chronic obstructive pulmonary disease in stable stage were random-ly divided into melatonin group and control group,and 21 patients in each group treated with melatonin(3 mg/d) or placebo for 3 months respectively. The plasma levels of 8- isoprotane,IL-8,TNF-α,h-CRP pulmonary func-tion,six minutes walking test and MRC dyspnea score before treatment,2 months and 3 months after the treatment were analyzed. Results After 2 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(10.40 ± 5.4 vs. 16.92 ± 4.33,P<0.05),and the concentration of IL-8 (6.88 ± 2.37 vs. 11.33 ± 3.39,P < 0.05). After 3 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(9.40 ± 4.0 vs. 17.92 ± 3.33,P < 0.01),and IL-8 (5.67 ± 3.22 vs. 9.31 ± 3.23,P < 0.05). Compared with before treatment,melatonin could significantly de-creased the concentration of 8-isoprotane(9.40 ± 4.0 vs. 20.40 ± 8.4,P<0.01 )and IL-8(5.67 ± 3.22 vs. 12.33 ± 3.88,P<0.05)after 3 months. Meanwhile,the concentration of the TNF-α(25.83 ± 9.18 vs. 35.83 ± 12.18,P<0.05)and hypersensitive C(1.76 ± 1.18 vs. 3.09 ± 1.79,P < 0.05)reactive protein in the melatonin group was greatly lower than the placebo group. After 3 months,compared to the placebo group,MRC dyspnea score of pa-tients in the group of melatonin was improved significantly(1.56 ± 1.38 vs. 2.09 ± 1.16,P<0.05 ),and lung func-tion and six minutes walk test showed no significant difference between patients in the two groups. Conclusions Exogenous melatonin administration can decrease the concentration of 8-isoprotane,IL-8,TNF-αand h-CRP in the blood of patients with moderate to severe COPD ,and improve the MRC dyspnea score. Melatonin has a significant effect on reducing oxidative stress and inhibiting inflammatory reaction in patients with moderate and severe stage stable COPD,which demonstrates its potential therapeutic value with broad clinical application prospects.
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Abstract Currently, there is a paucity of available treatment strategies for oxidative phosphorylation disorders. Coenzyme Q10 (CoQ10) and related synthetic quinones are the only agents to date that have proven to be beneficial in the treatment of these heterogeneous disorders. The therapeutic efficacy of CoQ10 is not restricted to patients with an underlying CoQ10 deficiency and is thought to result from its ability to restore electron flow in the mitochondrial respiratory chain (MRC) as well as to increase the cellular antioxidant capacity. At present, however, there is no consensus on the appropriate dosage or therapeutic plasma level of CoQ10, and this information will be required before CoQ10 can be utilized effectively in the treatment of mitochondrial disease. The following review will outline our current knowledge on the use of CoQ10 in the treatment of MRC disorders and primary CoQ10 deficiencies.
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OBJECTIVE: To identify the factors that could predict the functional outcome in patients with the axonal type of Guillain-Barre syndrome (GBS). METHODS: Two hundred and two GBS patients admitted to our university hospital between 2003 and 2014 were reviewed retrospectively. We defined a good outcome as being "able to walk independently at 1 month after onset" and a poor outcome as being "unable to walk independently at 1 month after onset". We evaluated the factors that differed between the good and poor outcome groups. RESULTS: Twenty-four patients were classified into the acute motor axonal neuropathy type. There was a statistically significant difference between the good and poor outcome groups in terms of the GBS disability score at admission, and GBS disability score and Medical Research Council sum score at 1 month after admission. In an electrophysiologic analysis, the good outcome group showed greater amplitude of median, ulnar, deep peroneal, and posterior tibial nerve compound muscle action potentials (CMAP) and greater amplitude of median, ulnar, and superficial peroneal sensory nerve action potentials (SNAP) than the poor outcome group. CONCLUSION: A lower GBS disability score at admission, high amplitude of median, ulnar, deep peroneal, and posterior tibial CMAPs, and high amplitude of median, ulnar, and superficial peroneal SNAPs were associated with being able to walk at 1 month in patients with axonal GBS.
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Humanos , Potenciales de Acción , Axones , Síndrome de Guillain-Barré , Estudios Retrospectivos , Nervio TibialRESUMEN
Background: Evaluation of jaundice patients should include proper history and examination, laboratory investigation and imaging investigations (non invasive like Ultrasound (U\S), CT and MRI or invasive like ERCP and PTC). Aim of Study: The aim of this prospective study is to evaluate the diagnostic reliability of U\S and MRI-MRCP in patients of obstructive jaundice in clinical practice. Materials and Methods: This is a prospective study performed on 60 patients (31 male and 29 female) with an average age of 55.53 +/- 17.57 years presented with obstructive jaundice for whom abdominal ultrasound (U\S) and magnetic resonance imaging (MRI) and magnetic resonance cholangiopancreatography (MRCP) on 3 Tesla was performed in the departments of radiology in Max super speciality teaching hospital, saket, Delhi, India from May 2012 to May 2013. The final diagnosis was achieved by endoscopic retrograde cholangiopancreatography (ERCP) and \or surgery and confirmed by histopathology. Results: The most common cause of obstructive jaundice in our study was common bile duct stones (51.65%) followed by tumors (33.3%) then benign strictures (10.0%), choledochal cyst (3.33%). In this study, MRI-MRCP could differentiate surgical from medical jaundice in all cases, while U\S could differentiate surgical from medical jaundice in 91.25% of cases. MRI-MRCP correctly defines the level of obstruction in all cases (100%). While U\S correctly define the level of obstruction in only 78% of the total cases. MRI-MRCP correctly suggests the most possible cause of obstruction in 96.25% of cases. While USG is correctly suggests the most possible cause in only 76.3%. Conclusion: So that USG as a screening modality is useful to confirm or exclude biliary dilatation & to choose patients for MRCP examination. MRI-MRCP is a useful non-invasive and essential method in the preoperative evaluation of patients with obstructive jaundice. In addition MRI-MRCP was superior to U\S or ERCP in studying the extent & staging of malignant lesions.
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Objective To optimize the culture conditions of MRC -5 human diploid cell.Methods To compare the growth status of MRC -5 cells,three kinds of culture medium with T25 bottles and Spinner cultivation system Cytodex1 micro carrier were used.Morphology,cell counting,growth curve,glucose -lactic acid value were observed and detected daily for screening a kind of suitable medium.Cell proliferation was compared with different levels of the bovine serum.Results There were no significant differences among the three kinds of culture medium.There were significant differences among MEM((43.25 ±0.60)×104 cells/mL,(12.98 ±1.27)×105 cells /mL),M199 ((35.40 ±1.41 )×104 cells/mL, (10.76 ±1.31)×105 cells /mL)and DMEM/F12 ((36.75 ±1.59)×104 cells/mL,(11.22 ±1.42)×105 cells /mL)(P<0.01).The cell proliferation of MEM cultures was 5.17 and 6.49 times better than those of M199 and DMEM/F12 cultures.Imported fetal bovine serum cell proliferation ((4.55 ±0.51)×105 cells /mL)was better than the other three bovine serum ((4.12 ±1.03,3.59 ±0.48,3.53 ±0.52)×105 cells /mL).Conclusion Tree kinds of culture medium can be used to culture MRC -5 human diploid cell.The MEMculture is better.Imported fetal bovine serum is better than other kinds of serum.
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El propósito de una medición es determinar el valor de una magnitud, llamada el mensurando. La imperfección natural de las mediciones, hace imposible conocer con certeza absoluta el valor verdadero de una magnitud, ya que toda medición lleva implícita una incertidumbre, el cual es un parámetro no negativo que caracteriza la dispersión de los valores atribuidos a un mensurando. En la presente investigación se empleo una estrategia diferente al procedimiento bottom-up propuesto por la ISO, el cual nos permitió estimar de forma global la incertidumbre, mediante la agrupación de valores determinados durante la verificación del método analítico. El valor obtenido en nuestro estudio fue de 3.46 UI/mL con un 90% de confianza. Observándose como fuente de mayor contribución al cálculo, el efecto de la matriz del medicamento, dependiente del tipo de insulina presente, además de la precisión y trazabilidad del patrón de referencia, los cuales contribuyen en la incertidumbre final de forma similar. En este sentido, nuestros esfuerzos deben dirigirse a la optimización de los procesos de homogeneización, adquisición del patrón de referencia tipo primario, con la finalidad de garantizar a lo largo del tiempo, resultados reproducibles, trazables y confiables.
The purpose of measurement is to determine the value of a quantity, called the measurand. The natural imperfection of determining measurements, makes impossible to know with absolute certainty the true value of a magnitude, since all measurement implies an uncertainty, which is a non-negative parameter that characterize the dispersion of the values attributed to a measurand. In this research, we employed a different strategy to the "bottom-up" procedure, proposed by the ISO, which allowed us to calculate globally the uncertainty, by grouping values, determined during the verification of the analytical method. The obtained value in our study was 3.46 IU/mL with 90% confidence. The matrix effect of the drug, depending on the insulin type present inside, was observed as the bigger contribution source to the calculation, as well as the accuracy and traceability of the reference pattern, which contribute in the final uncertainty similarly. Thus, our efforts should be directed to the optimization of the homogenization processes and the acquisition of the primary type reference pattern, in order to ensure over time, reproducible, traceable and reliable results.
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Humanos , Masculino , Femenino , Biotecnología/métodos , Cromatografía Liquida/instrumentación , Composición de Medicamentos , Insulina/administración & dosificación , Valores de Referencia , Salud PúblicaRESUMEN
OBJECTIVE: Since IVF program was first established, various types of media and culture systems have been developed either in-house or commercially. The aim of this study was to compare the efficacy of in-house Maria Research Center (MRC) media to that of commercially available Sydney IVF media in human day 3 embryo transfer cycles. METHODS: Three hundred sixty nine couples were included in this prospective, randomized, and comparative study. All couples undergoing IVF treatment at the Maria Fertility Hospital were randomly assigned to either Sydney IVF (n=178) or MRC (n=191) media. RESULTS: No difference was observed between the MRC media and Sydney IVF media groups with respect to fertilization rate (74.4% vs. 75.5%). The clinical pregnancy and implantation rates of MRC media (47.1% and 20.0%, respectively) were also similar to those of Sydney IVF media (44.4% and 19.4%, respectively). However, the proportion of embryos with good quality on day 3 was significantly higher in the MRC media group than the Sydney IVF media group (50.2% vs. 43.2%) (p<0.05). CONCLUSION: MRC media were as effective as Sydney IVF media for sustaining embryo development and pregnancy rates. The present study implies that MRC media can be a suitable alternative to commercially available media for human IVF-ET program.
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Femenino , Humanos , Embarazo , Transferencia de Embrión , Desarrollo Embrionario , Estructuras Embrionarias , Composición Familiar , Fertilidad , Fertilización , Índice de Embarazo , Estudios ProspectivosRESUMEN
ObjectiveStromal cell-derived factor -1 (SDF-1 ) is closely related to the biological characteristics of breast cancer. We aimed to explore whether estrogen affected breast cancer by SDF-1. MethodsThe breast cancer cell line MCF-7 and MRC5 were chosen, and divided into three groups: the control group, the estrogen group and the estrogen + estrogen receptor blocker group. Each group was cultured with different physiological concentrations of 17-β estrogen at certain time, and the same alcohol concentration of 17-β estradiol at different time points, and then the enzyme-linked immunosorbent assay (ELISA) was used to measure the concentration of SDF-1 in culture medium, and the semi-quantitative reverse transcriptionpolymerase chain reaction (RT -PCR) was used to detect the expression of SDF-1 mRNA in each group.ResultsSDF-1 can be detected in the culture medium of both MCF-7 and MRC5 cell lines. All different concentrations of 17-β estradiol may increase the secretion of SDF-1 in MCF-7 cells. When adding 17-β estradiol to the concentration of 107mol/L, the secretion of SDF-1 reached the peak in 2 hours, which was 6 times and 2.7 times that of control group ( P ＜ 0.01 ). The effect could be ehminated by pure estrogen receptor ICI182,780. In addition, the mRNA expression of SDF-1 was consistent with the SDF-1 protein levels-l07 mol/L group. The expression of SDF-1 mRNA was higher than both that of the control group and the blocking group in 2 hours (P ＜ 0.05 ). ConclusionsIn some breast cancer cell lines, physiological concentrations of estrogen can increase the secretion of SDF- 1, and this effect is mainly achieved through the estrogen receptor. Estrogen can influence the biological characteristics of breast cancer by SDF-1.
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OBJECTIVE: To compare muscle strength (MS) and motor function in patients with Duchenne muscular dystrophy (DMD) receiving steroids for different times against the natural evolution of DMD described by Scott et al. METHOD: 90 patients with DMD (aged 5- 12 years), receiving steroids for one to seven years, were evaluated by Medical Research Council Scale (MRC) and Hammersmith motor ability score. The relation between MS and motor abilities measurement from our data and Scott's ones were ascertained statistically. RESULTS: The relation between patient's age and Hammersmith scores revealed decrease of 0.76 point per year for age against decrease of 2.23 points on Scott's study. The relation between MRC scale and patient's age showed decrease of 0.80 point per year of age against decrease of 3.65 points on Scott's study. CONCLUSION: In patients with DMD aged five to 12 years the progression of the disease is delayed by steroids and the motor function is less reduced than muscular strength.
OBJETIVO: Comparar força muscular e função motora de pacientes com distrofia muscular de Duchenne (DMD) em corticoterapia com a evolução natural da doença descrita por Scott et al. MÉTODO: Noventa pacientes, entre 5 e 12 anos de idade, em corticoterapia por um até sete anos, foram avaliados quanto à força muscular (FM) (escala MRC) e função motora (Hammersmith motor ability score). A relação entre idade, FM e função motora e a comparação com o estudo de Scott et al foram determinadas estatisticamente. RESULTADOS: a relação idade/escore Hammersmith diminuiu 0,76 pontos a cada ano de aumento da idade (2,23 pontos na história natural). A relação idade/MRC decresceu 0,80 pontos a cada ano de aumento da idade (3,65 pontos na história natural). CONCLUSÃO: Nos pacientes em corticoterapia, a progressão da doença é mais lenta que na evolução natural em todas as faixas etárias avaliadas, sendo a FM mais comprometida que a função motora.