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ObjectiveTo investigate the effect of myosin heavy chain 7 gene-derived miRNA-208b-3p on the fibrotic phenotype of cardiac fibroblasts. MethodsmiRNA chip array was performed to detect the dysregulated miRNAs in the myocardium of diabetic db/db mice and db/m control mice. Neonatal mouse ventricular cardiomyocytes （NMVCs） and cardiac fibroblasts （CFs） were isolated from C57BL/6 mice and cultured. Real-time quantitative PCR （RT-qPCR） was conducted to determine the expression of miR-208b-3p in mouse CFs and NMVCs subjected to angiotensinⅡ（AngⅡ） and high glucose plus glucose oxidase （G/Go） treatment， respectively. Cell counting kit 8（CCk8） assay， flow cytometry and determination of fibrosis-related protein， including COL1A1， COL3A1and α-SMA， were performed in mCFs transfected with miR-208b-3p. Dual luciferase reporter assay was performed to confirm the interaction between miR-208b-3p and the 3'-UTR of metal response element binding transcription factor 2 （Mtf2） and progesterone receptor membrane component 1（Pgrmc1）， respectively. The expressions of Mtf2 and Pgrmc1 at the mRNA and protein levels in mCFs after miR-208b-3p mimic transfection were determined using RT-qPCR and Western blot assay， respectively. The small interfering RNA （siRNA） was used to inhibit Mtf2 and Pgrmc1 expression in mCFs， and the effects of Mtf2 siRNA， Pgrmc1 siRNA and miR-208b-3p on fibrosis-related protein expression in mCFs were investigated. ResultsResults of miRNA chip array and RT-qPCR assay showed that miR-208b-3p was up-regulated in the myocardium of the diabetic db/db mice. miR-208b precursor and the host gene of Myh7 were consistently increased in db/db mice. miR-208b-3p and Myh7 mRNA were expressed in mCFs and NMVCs， but the levels of miR-208b-3p and Myh7 mRNA in NMVCs were much higher than those in mCFs. miR-208b-3p was up-regulated in mCFs and NMVCs subjected to Ang Ⅱ and G/Go treatment， respectively. miR-208b-3p could significantly enhance fibrosis-related protein， including COL1A1， COL3A1 and α-SMA， in mCFs， without affecting the proliferation activity and cell cycle distribution of mCFs. Dual luciferase reporter assay revealed the interactions of miR-208b-3p with the 3'-UTR of Mtf2 and Pgrmc1. The results of RT-qPCR and Western blotting confirmed that miR-208b-3p inhibited Mtf2 and Pgrmc1 expression at the post- transcriptional level. Transfection with miR-208b-3p mimic， Mtf2 siRNA and Pgrmc1 siRNA could consistently enhance the fibrosis-related protein expression in the cardiac fibroblasts. ConclusionsmiR-208b-3p enhances fibrosis-related gene expression by targeting Mtf2 and Pgrmc1in mCFs.
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OBJECTIVE@#To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) on venous thrombosis and quality of life after lung cancer surgery, basing on the conventional nursing and early functional exercise.@*METHODS@#A total of 120 patients diagnosed as non-small cell lung cancer (NSCLC) and received radical resection of lung cancer surgery for the first time were randomized into a conventional nursing group, a rehabilitation training group and a TEAS group, 40 cases in each group. Conventional nursing was adopted in the conventional nursing group. Conventional nursing combined with early functional exercise were adopted in the rehabilitation training group, the exercise was taken 20 min each time, once in both morning and afternoon for 5 days. On the basis of the treatment in the rehabilitation training group, TEAS was applied at Zusanli (ST 36), Xuehai (SP 10), Sanyinjiao (SP 6), etc. in the TEAS group, with disperse-dense wave in frequency of 30 Hz/100 Hz and tolerable intensity, 30 min each time, once in both morning and afternoon for 5 days. The incidence of venous thrombosis in each group was observed at the 5th day after surgery. Before surgery and at the 5th day after surgery, the Caprini thrombus risk assessment was performed, the Karnofsky performance status (KPS) scale and the functional assessment of cancer therapy-lung (FACT-L) were used to evaluate the quality of life.@*RESULTS@#At the 5th day after surgery, no thrombosis was found in the TEAS group, the incidence of venous thrombosis in the TEAS group was lower than 15.0% (6/40) in the conventional nursing group (@*CONCLUSION@#On the basis of the conventional nursing and early functional exercise, TEAS can reduce the incidence of venous thrombosis, effectively prevent thrombosis and improve quality of life.
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Humans , Acupuncture Points , Carcinoma, Non-Small-Cell Lung/surgery , Lung Neoplasms/surgery , Quality of Life , Transcutaneous Electric Nerve Stimulation , Venous Thrombosis/etiologyABSTRACT
@#【Objective】To investigate the role and the potential target of miR-199a-3p in mouse cardiac hypertrophy.【Methods】Neonatal mouse ventricular cardiomyocytes（NMVC）were isolated from the hearts of 0- 3- day- old newborn C57BL/6 mice. MiR-199a-3p mimic and retinoblastoma transcriptional corepressor 1（Rb-1）siRNA were transfected in? to NMVC to elevate the level of miR-199a-3p and inhibit Rb-1 expression，respectively. NMVC were stained with FITC- phalloidin solution to determine the size of NMVC. Dual luciferase reporter assay was performed to identify the interaction between miR- 199a- 3p and the 3’UTR of Rb- 1. mRNA and protein expression of cardiac hypertrophy associated genes were determined by RT-qPCR and Western blotting assay，respectively.【Results】（1）Over-expression of miR-199a-3pcould significantly enhance the expression of cardiac hypertrophy-related genes in NMVC ；（2）Dual-luciferase reporter assay results verified that miR- 199a-3p can interact with the 3’UTR of Rb-1. MiR-199a-3p could suppress Rb-1 ex? pression at the post-transcriptional level；（3）Functionally，miR-199a-3p mimic，consistent with Rb-1 siRNA，could increase cell size and the expression of Nppa，Acta1 and Myh7 in NMVC，and promote the nuclear translocation of E2f2 in NMVC.【Conclusions】MiR-199a-3p promotes the entry of E2f2 into the nucleus through inhibiting the expression of Rb-1，contributing to cardiomyocyte hypertrophy.
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The drug delivery system with "gatekeeper" is designed to achieve a stable entrapment state of the payload under normal physiological conditions through the gatekeepers. With tumor microenvironment or stimulation of exogenous factors, the gatekeeper is detached or altered to promote the responsive release of the drug. In this paper, we focus on applications of stimuli-responsive linkages and stimuli-responsive groups, and review research progresses of drug delivery system with "gatekeeper" developed over the past 10 years.
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Objective:To observe the clinical effects of tuina plus Western medication for functional dyspepsia (FD) due to liver qi stagnation and spleen deficiency.Methods:A total of 72 patients in conformity with the inclusion criteria of FD were randomly divided into an observation group and a control group based upon the random number table,36 cases in each group.The control group was treated with mosapride citrate dispersible tablets,and the observation group was treated with the same tablets plus tuina.Before the treatment and 4 weeks after the treatment,the clinical symptoms,quality of life (QOL) and depression severity were observed by the scale,and were followed up two months later after the treatment for assessment of the clinical effects.Results:After the treatment and at the follow-up,the symptom scores of FD and the sores of Hamilton depression rating scale (HAMD) in both groups decreased,and the scores in Chinese version of quality of life questionnaire for functional digestive disorders (Chin-FDDQL) increased,with statistically significant differences in comparison with the same group before the treatment (all P＜0.05).In comparison between the two groups at the same time point after the treatment,the scores of FD symptoms,HAMD and Chin-FDDQL were improved better in the observation group than those in the control group,with statistically significant differences (all P＜0.05).The total effective rates at the follow-up were 91.7％ in the observation group and 75.0％ in the control group,without statistical difference between the two groups (P＞0.05).The rate of clinical cure and remarkable effect was 66.7％ in the observation group,higher than 41.7％ in the control group,it is higher in the observation group than that in the control group,with a statistically significant difference between the two groups (P＜0.05).Conclusion:Tuina plus Western medication is precise in the therapeutic effects for FD due to liver qi stagnation and spleen deficiency and can effectively relieve clinical symptoms,elevate the QOL and alleviate depression severity of the patients.Moreover,it's better than the treatment by Western medication alone in the long-term therapeutic effects.
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Objective: To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1 (TFF1) and mucin 5AC (MUC5AC) levels, as well as the expression of epidermal growth factor receptor (EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome. Methods: Seventy-five SPF grade Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a 3 moxa-cone ginger-partitioned moxibustion group (group C1), a 6 moxa-cone ginger-partitioned moxibustion group (group C2) and a 9 moxa-cone ginger-partitioned moxibustion group (group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ℃200% concentrated Da Huang (Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5AC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry. Results: After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased (all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased (all P<0.01). Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased (all P<0.01), however, there was no significant difference between group C2 and C3 (all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein. Conclusion: Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.
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The concept and principle of 3D printing technology were described,and its advantages were expounded according to the demands of emergency medical equipment.The application of 3D printing technology in the field of emergency medical equipment was analyzed from six aspects,such as equipment system,disposition model,development and innovation,use-pattern,maintenance methods,comprehensive benefit etc,to provide the references to the future construction and development of emergency medical equipment.It's pointed out 3D,4D and 5D printing technologies were of great significance for driving the development of emergency medical equipment.
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The concept and principle of 3D printing technology were described,and its advantages were expounded according to the demands of emergency medical equipment.The application of 3D printing technology in the field of emergency medical equipment was analyzed from six aspects,such as equipment system,disposition model,development and innovation,use-pattern,maintenance methods,comprehensive benefit etc,to provide the references to the future construction and development of emergency medical equipment.It's pointed out 3D,4D and 5D printing technologies were of great significance for driving the development of emergency medical equipment.
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To systematically review the efficacy and safety of Danhong injection for patients with idiopathic pulmonary fibrosis(IPF), two researchers electronically searched PubMed, EMbase, Web of Science, Cochrane Library, CNKI, CBM, WanFang Data and VIP databases from the date of establishment to May 2016 for all randomized controlled trials(RCTs) and quasi-RCTs on the use of Danhong injection in patients with IPF. Manual search in relevant journals and search of relevant literature on other websites were also performed. The data extraction and quality assessment of included RCTs and quasi-RCT were conducted by two reviewers independently. Then, Meta-analysis was conducted by using RevMan 5.3 software. A total of 12 RCTs involving 844 patients were included, 423 cases in experiment group and 421 cases in control group. The results of meta-analysis indicated that the Danhong injection group was superior than the control group in clinical effectiveness(RR=1.36, 95%CI 1.25 to 1.49, P<0.000 01), increased DLCO value(MD=4.25, 95%CI 3.32 to 5.18, P<0.000 01), and increased PaO2 value(MD=14.51, 95%CI 12.35 to 16.68, P<0.000 01). The analysis results showed that Danhong injection could significantly reduce the level of TGF-β1 in serum. There were no serious or frequently happened adverse effects in the Danhong injection group, indicating high safety and good tolerance of Danhong injection in treatment of IPF. The current evidences suggested that Danhong injection in short term use(<12 weeks) could increase clinical effectiveness, improve DLCO and PaO2, and decrease the level of TGF-β1 in serum of IPF patients, with less adverse effects. However, these results should be carefully interpreted due to the low methodology quality and small sample size of trials, and this conclusion had to be further verified by high quality, large scale and double blinded RCTs.
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OBJECTIVE: To study on characterization, irritation, the release mechanism and the elimination kinetics of Fraxini Cortex thermosensitive in-situ-forming eye gel (FC-ISG). METHODS: The non-membrane dissolution model was used to observe the release mechanism of FC-ISG. The stabilities of FC-ISG were investigated under following circumstances bright light, freeze test and accelerating test. Single-dose and multiple-dose irritations of FC-ISG were evaluated by draize test. The elimination kinetics of FC-ISG were analyzed by non-compartment model. RESULTS: FC-ISG showed good stability and non-stimulation to rabbit eyes. Drug release from FC-ISG was completely controlled by gel erosion, the release kinetics was coincided with zero-level release. AUC and MRT in FC-ISG group were significantly higher than those in control group (P<0.05). CONCLUSIONS: FC-ISG can improve the bioavailability of drug by prolonging the residence retention time of drug in cornea. FC-ISG shows a great potential in ocular application.
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OBJECTIVE: To study the effects of 5-Fu combined with resveratrol (Res) on the growth and apoptosis of A431 and TE-1 cell lines and the underlying mechanism. To study the therapeutic effects of 5-Fu/Res combination on mouse skin papilloma chemically induced by DMBA/TPA. METHODS: The effects of 5-Fu/Res combination on the viability of cancer cells were evaluated by MTT assay, growth curves assay and LDH releasing assay. The inhibitory effect of combination of the two drugs was analyzed by the method of Chou and Talalay. Apoptoses of A431 and TE-1 were determined by inverted microscope and gel electrophoresis of DNA fragment analysis. Intracellular Ca2+ concentration was determined to study the underlying mechanisms of 5-Fu and Res on the apoptosis of cancer cells. The mouse skin papilloma model was established by DMBA and TPA, the expression of actived-caspase-3 in mouse skin was examined by immunohistochemistry. RESULTS: Combination of 5-Fu and Res decreased the median inhibitory concentration (IC50) to cancer cells remarkably. 5-Fu/Res combination showed a synergistic effect on apoptosis of A431 and TE-1 cells. Much more typical morphological changes of apoptosis and amount of fragmented DNA were observed in the cells treated with 5-Fu and resveratrol in combination than that in the cells treated with the agents alone. The increase of [Ca2+]i induced by 5-Fu/Res combination might be involved in the apoptotic induction. There was a significant decrease of the number of tumors after treatment with 5-Fu and Res in the tumor-bearing mice model. Active caspase-3 in the cells of mice skin was generated by 5-Fu in combination resveratrol was more effective than either alone. CONCLUSION: The 5-Fu/Res combination shows synergistic anti-tumor effects both in vitro and in vivo.
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Objective To study protective effect and mechanism of caffeic acid phenethyl ester ( CAPE ) in rats with doxorubicin-induced myocardial injury.Methods Sixty male SD rats were randomly divided into control group, model group and CAPE ( 12、24、48 mg/kg ) groups.The models of doxorubicin-induced myocardial injury were established by injection of 2.5 mg/kg doxorubicin every other day for 6 times.After the last injection, model group was given distilled water and CAPE ( 12、24、48 mg/kg ) groups were fed with CAPE, one time a day for 8 weeks.After consecutive 8 weeks, serum creatine kinase, lactate dehydrogenase and BNP were assessed , NO concentration, MDA,GSH,SOD,NOS,LPO and CAT contents in the heart were determined and histopathological changes were detected.Results serum creatine kinase、lactate dehydrogenase and BNP of CAPE 48 mg/kg group decreased significantly(P<0.05), while the activity of NO,NOS,GSH,CAT and SOD increased in the CAPE 48 mg/kg group significantly ( P<0.05 ) .Histopathology showed caffeic acid phenethyl ester could improve the doxorubicin-induced myocardium injury. Conclusion caffeic acid phenethylester has the protective effects in rats with doxorubicin-induced myocardial injury.Protective effect may be related to improve myocardial against oxidative stress damage.
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Cancer metastasis is the most dangerous stage of tumorigenesis and evolution, the primary cause of death in cancer patients. Clinically, more than 60% of cancer patients have found metastasis at the time of examination. Modern medicine has made significant progress on the mechanisms of cancer metastasis in recent years, from the simple "anatomy and machinery" theory forward to the "seed and soil" theory, then to the "microenvironmental" theory and the "cancer stem cell" theory. The emerging "cancer stem cell" theory successfully explains phenomenon such as tumor genetic heterogeneity, anoikis resistance, tumor dormancy, providing more new targets and ideas for the diagnosis and treatment of cancer metastasis.
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Animals , Humans , Medicine , Methods , Neoplasm Metastasis , Neoplasms , Drug Therapy , PathologyABSTRACT
Objective To explore Chongqing migrant workers Suiqian children′s mental health status and related factors .Meth-ods In this study 740 children from grade 7 to grade 9 in 3 schools which specify recruit migrant workers′children ,including the 399 migration children of migrant workers and 341 urban household registration students were involved .They were investigated with symptom rating scale(SCL-90) and self-made general questionnaire .Results There were no significant differences in SCL-90 scores between the migrant workers migration children group and urban household registration students group (P>0 .05) .Conclu-sion Generally speaking ,Chongqing migrant workers Suiqian children′s mental health status is good ,but there are still some prob-lems .We need do some further study on the mode of psychological intervention to maintain their physical and mental health .
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<p><b>BACKGROUND</b>Everolimus, a derivative of sirolimus, is a potent immunosuppressant that has important anti-proliferative properties. In the present study, we demonstrated the inhibiting neointimal hyperplasia in injured carotid arteries in rats by using two different doses of everolimus administrated via the oral route for a long time.</p><p><b>METHODS</b>A rat model of carotid artery injury was established by balloon inflation. Eighty rats were randomly divided into the sham-operated group (n = 20), injury group (n = 20), low dosage of everolimus group (n = 20), and high dosage of everolimus group (n = 20). The low dose of everolimus (1.5 mg/kg) was given one day before injuring the carotid artery by balloon, followed by 0.75 mg/kg per day for 28 days via intragastric gavage. High dose everolimus (2.5 mg/kg) was given one day before injuring the carotid artery by balloon, followed by 1 mg/kg per day for 28 days. Expression of eukaryotic translation initiation factor 4E (eIF-4E) and phosphorylation of ribosomal protein S6 kinase 1 (P70S6K) were determined by reverse transcription-polymerase chain reaction and Western blotting analysis.</p><p><b>RESULTS</b>In the injured carotid artery, neointimal hyperplasia was normally observed four weeks after injury. Everolimus inhibited neointimal hyperplasia after balloon injury in a dose dependent manner. At the same time, the study demonstrated that everolimus reduced the expression of P-P70S6K, eIF-4E, transforming growth factor (TGF)-β1 and of proliferating cell nuclear antigen (PCNA).</p><p><b>CONCLUSIONS</b>Everolimus significantly inhibited neointimal hyperplasia of the injured carotid artery. The effect depended on dosage and was associated with the reduction of phosphorylation of P70S6K and the eIF-4E expression level.</p>
Subject(s)
Animals , Male , Rats , Carotid Arteries , Carotid Artery Injuries , Drug Therapy , Carrier Proteins , Metabolism , Everolimus , Neointima , Drug Therapy , Phosphoproteins , Metabolism , Phosphorylation , Rats, Sprague-Dawley , Ribosomal Protein S6 Kinases, 70-kDa , Metabolism , Sirolimus , Therapeutic UsesABSTRACT
<p><b>BACKGROUND</b>The effect of impaired glucose tolerance (IGT) on cardiac function during the chronic prediabetes state is complicated and plays an important role in clinical outcome. However, the molecular mechanisms are not fully understood. This study was designed to observe cardiac dysfunction in prediabetic rats with IGT and to determine whether glucose metabolic abnormalities, inflammation and apoptosis are linked to it.</p><p><b>METHODS</b>The IGT rat models were induced by streptozocin, and the heart functions were assessed by echocardiography. Myocardial glucose metabolism was analyzed by glycogen periodic acid-Schiff staining, and the pro-apoptotic effect of IGT was evaluated by TUNEL staining. Additionally, caspase-3 activation, macrophage migration inhibitory factor (MIF) and G-protein coupled receptor kinase 2 (GRK2) were detected by Western blotting in cardiac tissue lysates.</p><p><b>RESULTS</b>Area-under-the-curve of blood glucose in rats injected with streptozotocin was higher than that in controls, increased by 16.28%, 38.60% and 38.61% at 2, 4 and 6 weeks respectively (F = 15.370, P = 0.003). Abnormal cardiac functions and apoptotic cardiomyocytes were observed in the IGT rats, the ejection fraction (EF) being (68.59 ± 6.62)% in IGT rats vs. (81.07 ± 4.59)% in controls (t = 4.020, P = 0.002). There was more glucose which was converted to glycogen in the myocardial tissues of IGT rats, especially in cardiac perivascular tissues. Compared to controls, the cleaved caspase-3, MIF and GRK2 were expressed at higher levels in the myocardial tissues of IGT rats.</p><p><b>CONCLUSIONS</b>IGT in the prediabetes period resulted in cardiac dysfunction linked to abnormal glycogen storage and apoptosis. Additionally, MIF and GRK2 may be involved in the pathogenesis of cardiac dysfunction in prediabetes and their regulation may contribute to the design of novel diagnostic and therapeutic strategies for those who have potential risks for diabetic cardiovascular complications.</p>
Subject(s)
Animals , Rats , Apoptosis , Blotting, Western , Disease Models, Animal , Echocardiography , G-Protein-Coupled Receptor Kinase 2 , Metabolism , Glucose Intolerance , Glucose Tolerance Test , In Situ Nick-End Labeling , Intramolecular Oxidoreductases , Metabolism , Macrophage Migration-Inhibitory Factors , Metabolism , Myocardium , Metabolism , Pathology , Myocytes, Cardiac , Pathology , Streptozocin , ToxicityABSTRACT
Objective: To evaluate the effect and safety of low-dose naloxone combined with sufentanil and ropivacaine for postoperative patient-controlled epidural analgesia (PCEA) in elderly patients undergoing total hip replacement. Methods: A total of 60 elderly patients (ASA I-II) undergoing total hip replacement were randomly assigned to two equal groups: Group C was given 0.5 μg/ml sufentanil in 0.15% ropivacaine; Group N was given the same solution with 0.09 μg/(kg·ml) naloxone. The 2 groups were followed up in a double-blinded manner: the visual analogue scale (VAS) for pain score was recorded at 2, 6, 12, and 24 h to evaluate the analgesia effect. Meanwhile, the incidences of postoperative nausea and vomiting (PONV), pruritus, sedation, respiratory depression, and hypotension were also recorded. Results: The VAS pain score in Group N was significantly lower than that in Group C at 6, 12 and 24 h (P<0.01). The incidence of PONV in Group N was significantly lower than that in Group C (P<0.05); the incidences of other opioid-induced side-effects were not significantly different between the 2 groups. During the course of analgesia, the vital signs of 2 groups were stable; no patients had respiratory depression or hypotension. Conclusion: Low-dose epidural naloxone can enhance the analgesic effect of sufentanil. With timely postoperative monitoring, low-dose naloxone combined with sufentanil and ropivacaine is safe and effective for postoperative patient-controtled epidural analgesia in elderly patients undergoing total hip replacement.
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<p><b>OBJECTIVE</b>To investigate the interaction domains between macrophage migration inhibitory factors (MIF) and the extracellular segment of type-II trans-membrane protein CD74 using a yeast two-hybrid system.</p><p><b>METHODS</b>By using molecular cloning techniques, the DNA fragments encoding MIF, MIF(50-65) and MIF(1-50/65-115) were introduced into the pGBKT7 vector to construct the corresponding recombinant bait plasmids, and the DNA fragments encoding CD74(73-232), CD74(73-109), CD74(1109-149) and CD74(149-232) into the pGADT7 vector to construct the recombinant activation domain (AD) plasmids. PEG/LiAC method was employed to transform the above 3 recombinant bait plasmids paired with each of the 4 recombinant AD plasmids into the chemical competent yeast AH109 cells. The transformed yeast AH109 cells were screened consecutively on SD/-Trp-Leu and SD/-Trp-Leu-Ade-His/X-alpha-gal nutritional media.</p><p><b>RESULTS</b>The results of restriction endonuclease digestion and DNA sequencing verified the correct construction of all the recombinant plasmids. The yeast AH109 cells transformed with each of the 3 recombinant bait plasmids could grow on SD/-trp nutritional media without autonomous activation effect on the reporter gene MEL1. The cells transformed with each of the 4 recombinant AD plasmids could also grow on SD/-leu nutritional media without activation of the reporter gene MEL1. Only the yeast AH109 cells co-transformed with MIF, MIF(50-65), or MIF(1-50/65-115) plasmid and CD74(73-232) plasmid could grow on SD/-Trp-Leu-Ade-His nutritional media with transcription activation of the reporter gene MEL1.</p><p><b>CONCLUSION</b>MIF interacts with the intact extracellular segment of CD74 (CD74(73-232)) independent of the functional domain of MIF(50-65).</p>
Subject(s)
Antigens, Differentiation, B-Lymphocyte , Genetics , Metabolism , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Extracellular Matrix , Metabolism , Histocompatibility Antigens Class II , Genetics , Metabolism , Macrophage Migration-Inhibitory Factors , Genetics , Metabolism , Peptide Fragments , Genetics , Protein Interaction Domains and Motifs , Genetics , Recombinant Proteins , Genetics , Two-Hybrid System TechniquesABSTRACT
<p><b>OBJECTIVE</b>To investigate the impact of antihypertensive medication timing on degree and stability of blood pressure (BP) lowering in patients with moderate and severe essential hypertension.</p><p><b>METHODS</b>Ninety patients were randomly assigned to take Valsartan and Felodiping together in the morning (group A), Valsartan in the morning and Felodiping in the evening (group B) or Felodiping in the morning and Valsartan in the evening (group C, n = 30 each). The morning dosage was titrated if the goal blood pressure was not achieved. Ambulatory blood pressure monitoring (ABPM) was performed on the first and 14(th) day of medication.</p><p><b>RESULTS</b>The BP reductions during nighttime and twenty-four in group B and C hours were similar (P > 0.05) but were significant more than those in group A (P < 0.05). The smoothness indexes of mean systolic, mean arterial blood pressure during nighttime and twenty-four in group B and C were similar but significantly higher than that in group A (P < 0.05). The smoothness index of diastolic pressure at nighttime in group B and C was similar but significantly higher than that in group A (P < 0.05).</p><p><b>CONCLUSION</b>More significant and stable antihypertensive effects could be achieved by taking the two antihypertensive medications separately in the morning and at evening compared that taken the two drugs together in the morning.</p>