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Dynamic variations of the cell microenvironment can affect cell differentiation, cell signaling pathways, individual growth, and disease. Optogenetics combines gene-encoded protein expression with optical controlling, and offers a novel, reversible, non-invasive and spatiotemporal-specific research tool to dynamically or reversibly regulate cell signaling pathways, subcellular localization and gene expression. This review summarizes the types of optogenetic components and the involved cellular signaling pathways, and explores the application and future prospects of the light-controlled cell signaling pathways.
Subject(s)
Cell Differentiation , Light , Optogenetics , Proteins , Signal TransductionABSTRACT
Although still in its infant stage, synthetic biology has achieved remarkable development and progress during the past decade. Synthetic biology applies engineering principles to design and construct gene circuits uploaded into living cells or organisms to perform novel or improved functions, and it has been widely used in many fields. In this review, we describe the recent advances of mammalian synthetic biology for the treatment of diseases. We introduce common tools and design principles of synthetic gene circuits, and then we demonstrate open-loop gene circuits induced by different trigger molecules used in disease diagnosis and close-loop gene circuits used for biomedical applications. Finally, we discuss the perspectives and potential challenges of synthetic biology for clinical applications.
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Objective To investigate the effect of the isotretinoin combined with Dazhui acupoint bloodletting in the treatment of adolescent acne.Methods78 patients with adolescent acne from department of dermatology in our hospital were selected and randomly divided into the control group and the experiment group,39 cases in each group.The control group were treated by isotretinoin and the experiment group were treated on the base of the control group with Dazhui acupoint bloodletting.The clinical total effective rate, incidence of adverse reactions and recurrence rate after 6 months were compared between control group and experiment group.ResultsCompared with the control group after treatment, the clinical total effective rate were higher(P<0.05), the incidence of adverse reactions were lower(P<0.05), the 6 months after the recurrence rate of acne were lower(P<0.05).ConclusionThe application of different dimension an acid combined with Dazhui acupoint bloodletting treatment can improve the adolescent acne clinical total efficiency, reduce the incidence of adverse reaction rate and recurrence rate, and suitable for clinical, it is worth popularization and application.
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Objective To study the influence of high quality nursing mode on the quality of life in endoscopic retrograde cholangiopancreatography (ERCP) patients. Methods A total of 292 patients with ERCP were randomly divided into control group and observation group with 146 cases in each group by the randomized block design method. The control group was given the conventional nursing care. The observation group was given the high quality nursing care based on the conventional nursing care. The score of Quality of Life of the World Health Organization (WHOQOL- 100), Quality of Life Index of Gastrointestinal Tract (GIQLI), Hamilton Anxiety Scale (HAMA), Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale (SDS), Visual Analogue Scale(VAS) before and after nursing care were observed and compared. The difference impact of two kinds of nursing mode on nursing quality between two groups was evaluated. Results WHOQOL-100 score in the observation group after treatment, including physiology and psychology and environment and social relations state such as grading, was respectively (59.3 ± 6.0), (61.8 ± 7.2), (60.3 ± 6.2), (62.8 ± 7.3) points and GIQLI index included self-conscious symptom and body physiological status and daily and social activities,such as state of emotional and psychological and the overall score, was respectively (79.8±4.9), (19.8±1.8), (14.7±1.9), (19.2±2.8), (105.7±6.6) points, which was (50.9±6.3), (52.5±6.7), (51.4±5.6), (53.4±7.1), (67.2±4.8), (15.6±1.9), (10.2±1.8), (16.3±2.3), (94.4 ± 6.2) points in control group, the difference between two groups was statistically significantly(t=1.876-2.327, P<0.05). HAMA score and SAS scores and SDS score and VAS score in the observation group after nursing was respectively (24.28 ± 4.78), (29.48 ± 6.54), (30.55 ± 7.32), (4.55 ± 1.18) points and respectively (36.68 ± 5.39), (41.72 ± 6.03), (42.65 ± 7.21), (6.07 ± 1.17) points in the control group, the difference between two groups was statistically significantly(t=2.876-4.012, P<0.05). Conclusions High quality nursing mode of ERCP in patients with perioperative patients of psychological and physiological all have different degrees of improvement and is suitable for popularization and application in clinic.
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Objective To analyze the clinical efficacy of desloratadine citrate disodium and fexofenadine in the treatment of refractory urticaria . Methods 100 patients with refractory urticaria were randomly divided into observation group and control group according to the digital table method, with 50 cases in each group.The control group were treated with desloratadine citrate disodium, and the observation group were treated with fexofenadine on the basis of control group.The efficacy, immunity function ( CD3 +, CD4 +, CD4 +/CD8 +) , the changes of inflammatory factors [ procalcitonin ( PCT) , C-reactive protein ( CRP) ] and the quality of life were compared between the two groups.The adverse reaction rate in two groups was recorded in two groups.Results The total efficacy in observation group (88.0%) was significantly higher than that in control group (72.0%)(P<0.05).The indicators of immunity function in observation group were significantly higher than those in control group(P<0.05).The inflammatory factors in observation group were significantly lower than those in control group ( P<0.05 ) .The sleep and exercise quality rate of the observation group were significantly higher than those in control group(86.0% vs.68.0%,84.0% vs.66.0%,P<0.05).Conclusion The combination of desloratadine citrate disodium and fexofenadine in the treatment of refractory urticaria has a good effect, can effectively improve the immune function, reduce the secretion of inflammatory factors, thus effectively relieve the clinical symptoms, improve their quality of life, and has high security.
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Objective To establish a HPLC method for the content determination of irbesartan in drugs.Methods The diamonsil C18 column (150 mm ×4.6 mm,5μm) was used, with the mobile phase of phosphoric acid solution-acetonitrile(adjusted to pH 3.2 with triethylamine)(62:38).The column temperature was 35℃, with the UV detection wavelength of 245 nm and a flow rate of 1.0 mL/min.Results Irbesartan exhibited a good linear relationship with the peak area in the concentration range of 0.04-0.40 mg/mL with the correlation coefficient of 0.999 99.The average recovery was 98.37% with RSD of 0.7%(n=9).Conclusion This method is simple and accurate with wide linear range and good reproducibility,and can be used to determine the content of irbesartan in drugs.
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Background Our previous studies found that mesenchymal stem cells (MSCs) can ameliorate experimental autoimmune uveitis (EAU) and reduce tissue impairment.Its mechanism is still pending.Objective This study was performed to investigate the effects of MSCs on T cell subsets and antigen presenting cells (APCs) in EAU rats.Methods MSCs were isolated from bone marrow of six male Wistar rats and cultured by plastic adherence method.Twelve female Lewis rats were assigned randomly into MSCs group and PBS group.EAU rat model was induced by immunization with 200 μl emulsion containing 30 μg interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 polypeptide fragment R16 and complete Freund adjuvant (CFA).The eye manifestations of the rats were observed and scored under the slit lamp microscope after modeling.The R16-immunized rats were treated intravenously with 5×106/ml MSCs for 3 consecutive days from day 9 to 11 after modeling in the MSCs group,and the equivalent volume of PBS was used with the same way in the PBS group.Fifteen days after modeling,the spleens and draining lymph nodes were collected to evaluate the proportion of interferon-γ (IFN-γ) positive CD4+ T cells,interleukin-17 (IL-17)positive CD4+ T cells and forkhead helix transcription factor p3 (Foxp3) positive CD4+ T cells by flow cytometry.The T cells and APCs from the different groups were cocultured and divided into PBS cocultured group,MSCs cocultured group, PBS-MSCs cross-cultured group and MSCs-PBS cross-cultured group under the stimulation of R16 at the concentration of 0.3,1.0 or 10.0 μg/ml, and the proliferation indexes of the T cells in different groups were assayed by 5-bromodeoxyuridine (BrdU) Elisa kit.The use of experimental animals complied with the regulations on the management of experimental animals promulgated by the national science and technology commission.Results The ocular surface inflammatory scores of 11,12,13 and 14 days after modeling in the MSCs group were significantly lower than that in the PBS group (t=3.825,5.100,4.250,3.400, all at P＜0.05).Compared with the PBS group, the proportions of IFN-γ positive CD4+ T cells in spleen and draining lymph notes were considerably decreased in the MSCs group (t =5.651,4.376, both at P＜0.05) , so were the IL-17+ CD4+ T cells (t =3.300,4.925, both at P＜0.05).However,the proportions of Foxp3 + CD4+ T cells in spleen and draining lymph notes were statistically raised in the MSCs group compared with the PBS group (t =-5.172,-2.825,both at P＜0.05).The proliferation index of T cells increased with the rise of R16 dose in the PBS cocultured group, and the proliferation indexes were all declined in the MSCs cocultured group compared with the PBS cocultured group under the stimulation of 0.3,1.0 and 10.0 μg/ml of R16 (P =0.027,0.000,0.000).In addition, significant reduces of proliferation indexes of T cells were seen in the PBS-MSCs cross-cultured group and MSCs-PBS cross-cultured group in comparison with the PBS cocultured group when stimulated by 1.0 μg/ml and 10.0 μg/ml R16 (1.0 μg/ml R16 : P =0.001,0.000;10.0 μg/ml R16:P=0.000,0.000).Conclusions MSCs can ameliorate EAU by inhibiting the functions of antigen-specific T cells and APCs and up-regulating T regulatory cells in EAU rats.
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This study was aimed to discuss the distribution and protein expression level ofμ-opioid receptor (MOR) in hippocampus of premenstrual syndrome (PMS) liver-qi stagnation rat model, in order to initially reveal the action mechanism of PMS liver-qi stagnation and intervention effect ofShu-Yu (SY) capsule. Chronic restraint stress method was used to copy PMS liver-qi stagnation rat model.SY capsule ofTiao-Gan prescription was given as intervention. Immunofluorescence (IF) and western blot (WB) technique were used to detect MOR in hippocampal CA1 and CA3 brain area of rats from each group. The results showed that compared with the normal group, the hippocampus MOR distribution arrangement was messy with increased protein concentration in the model group (P< 0.01). After drug intervention, the MOR protein level returned to normal level. It was concluded that the pathogenesis of PMS liver-qi stagnation may be associated with high expression of MOR in hippocampus CA1 and CA3 region of rats. SY capsule can effectively correct and restore it to nearly normal level. It may be one of the central mechanisms in SY capsule treatment of PMS liver-qi stagnation.
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To establish a prokaryotic expression and purification protocol for nuclease P1 (NP1), we first obtained a synthetic NP1 by splicing 22 oligonucleotides with overlapping PCR. We constructed and transformed a secretory expression vector pMAL-p4X-NP1 into Escherichia coli host strains T7 Express and Origami B (DE3) separately. Then, the recombinant NP1 was purified by amylose affinity chromatography, and its activity, thermo-stability and metal-ion dependence were investigated systematically. The results indicated that the expressed fusion proteins MBP-NP1 (Maltose binding protein-NP1) existed mainly in soluble form both in host strains T7 Express and Origami B (DE3), but the specific activity of recombinant protein from Origami B(DE3) strain was higher than T7 Express strain (75.48 U/mg : 51.50 U/mg). When the MBP-tag was cleaved by protease Factor Xa, the specific activity both increased up to 258.1 U/mg and 139.2 U/mg. The thermal inactivation experiments demonstrated that the recombinant NP1 was quite stable, and it retained more than 90% of original activity after incubated for 30 min at 80 degrees C. Zn2+ (2.0 mmol/L) could increase enzyme activity (to 119.1%), on the contrary, the enzyme activity was reduced by 2.0 mmol/L Cu2+ (to 63.12%). This research realized the functional expression of NP1 in the prokaryotic system for the first time, and provided an alternative pathway for NP1 preparation.