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Objective To explore the mechanism of phlegm-resolving and stasis- removing herbals on NAFLD by observing expressions of PGC1α mRNA and insulin resistance. Methods A total of 60 male SD rats were randomly divided into the normal group, model group, positive medication control group, high-dose, middle-dose and low-dose group. The rats were fed with high-fat forage for 8 weeks. The positive medication control group were gavaged with Dongbao-Gantai liquid (0.9 g/kg/day), the high-dose, middle-dose and low-dose group were gavaged with Xiaotan-Huayu liquid (43.34、32.50、21.67 g/kg/day), and normal group, model group were gavaged with equal volume of distilled water. The drugs were given by 1 ml/100 g and last for 8 weeks. The levels of TC, TG, FFA, ALT, AST, FBG, FINS, and HOMA-IR in serum, and levels of TC, TG, and PGC-1α mRNA and pathological morphological changes in hepatic tissue were observed after 8 weeks. Results The levels of TG (0.55 ± 0.10 mmol/L, 0.58 ± 0.09 mmol/L, 0.67 ± 0.11 mmol/L vs. 1.18 ± 0.15 mmol/L), TC (1.48 ± 0.24 mmol/L, 1.69 ± 0.27 mmol/L, 1.74 ± 0.27 mmol/L vs. 3.29 ± 0.26 mmol/L), FFA (251.08 ± 48.18 μmol/L, 277.53 ± 56.73 μmol/L, 291.82 ± 48.67 μmol/L vs. 432.19 ± 67.83 μmol/L), ALT (29.32 ± 4.17 U/L, 31.26 ± 4.74 U/L, 33.56 ± 5.18 U/L vs. 47.21 ± 8.67 U/L), AST (11.05 ± 2.18 U/L, 12.15 ± 2.67 U/L, 12.96 ± 2.93 U/L vs. 19.43 ± 3.68 U/L), FBG (5.68 ± 1.22 mmol/L, 6.86 ± 1.36 mmol/L, 7.94 ± 1.82 mmol/L vs. 11.88 ± 2.54 mmol/L), FINS (8.48 ± 1.22 mmol/L, 9.55 ± 1.95 mmol/L, 9.96 ± 1.74 mmol/L vs. 12.96 ± 2.67 mmol/L), HOMA-IR (1.91 ± 0.26, 2.91 ± 0.65, 3.52 ± 0.58 vs. 6.89 ± 1.21) in serum of high-dose, middle-dose and low-dose groups were decreased than model group. Levels of FFA (242.19 ± 35.13 μmol/L, 259.78 ± 29.33 μmol/L, 277.62 ± 34.29 μmol/L vs. 436.48 ± 52.15 μmol/L), TG (23.65 ± 3.28 mmol/L, 24.41 ± 3.15 mmol/L, 25.37 ± 3.59 mmol/L vs. 15.98 ± 2.37 mmol/L), TC (7.15 ± 0.82 mmol/L, 8.60 ± 0.95 mmol/L, 8.86 ± 1.04 mmol/L vs. 36.98 ± 4.28 mmol/L) were in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly lower than the model group. The levels of PGC-1α mRNA (1.24 ± 0.06, 1.02 ± 0.07, 0.99 ± 0.08 vs. 0.43 ± 0.06) in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly higher than model group. Conclusions The phlegm-resolving and stasis-removing herbals may improve lipid metabolism by regulating the expression of PGC-1α mRNA, inhibiting gluconeogenesis and liver sugar output, correcting disturbance of lipid metabolism and improving insulin resistance.
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Objective To investigate the efficacy of massage combined with microwave therapy in treating Children's bronchial pneumonia with syndrome of deficiency of lung-spleen Qi. Methods Two hundred forty-four bronchial pneumonia Children were selected in our hospital and randomly divided into the control group and the observation group with 122 cases in each group referring to random number table. The control group was administered with the routine treatment. Based on the control group, the observation group received the massage and microwave therapy. The course was 2 week for both groups. The syndrome scores of traditional Chinese medicine were performed before and after treatment, and the disappearance of gasping, antipyretic, disappearance of moist rale in the lungs, and absorption time of chest X-ray were observed and recorded. the flow cytometry was used to detect peripheral blood levels of CD4+ and CD8+, and serum levels of IL-6 and IL-8 were detected by ELISA. The clinical effect rate was evaluated. Results The total effective rate was 97.5% (118/121) in the observation group and 91.7% (110/120) in the control group,with statistically significant differences (χ2=4.406, P=0.044). After treatment, the scores of cough, phlegm, shortness of breath, pale face, fatigue, spontaneous perspiration, poor appetite in the observation group were significantly lower than those in the control group (t value were 10.008, 11.529, 11.778, 12.043, 11.309, 10.128, 11.008,P<0.01), and elapsed time of gasping, antifebrile time, disappear time of lung moist crackles, absorption time were significantly earlier than the control group (t value were 15.121, 10.099, 18.441, 14.310, P<0.01). After 2 week’ treatment, the scores of symptoms of deficiency of lung-spleen Qi in the observation group were significantly lower than those of control group (P<0.01). Compared with the control group, the duration of wheezing and antipyretic, duration of lung dampness and chest X-ray absorption in the observation group were significantly decreased (P<0.05). The total effective rate of the observation group was 97.52%, which was significantly higher than the control group 92.67% (P<0.05). After 2 week treatment, the CD4+(41.03% ± 4.55% vs. 33.52% ± 3.95%, t=14.011) in the observation group was significantly higher, CD8+ (22.71% ± 2.83% vs. 26.65% ± 3.05%, t=10.061) was significantly lower than the control group, serum levels of IL-6 (28.13 ± 3.63 ng/L vs. 40.09 ± 4.61 ng/L, t=14.890) and IL-8 (21.41 ± 2.75 ng/L vs. 29.05 ± 3.61 ng/L, t=11.439) were significantly lower than the control group (P<0.01). Conclusions Massage combined with microwave therapy for the Children's bronchial pneumonia with deficiency of lung-spleen Qi syndrome can promote the improvement of symptoms, enhance the efficacy, increase the immune function and reduce inflammation level.
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Objective@#To investigate the efficacy of massage combined with microwave therapy in treating Children's bronchial pneumonia with syndrome of deficiency of lung-spleen Qi.@*Methods@#Two hundred forty-four bronchial pneumonia Children were selected in our hospital and randomly divided into the control group and the observation group with 122 cases in each group referring to random number table. The control group was administered with the routine treatment. Based on the control group, the observation group received the massage and microwave therapy. The course was 2 week for both groups. The syndrome scores of traditional Chinese medicine were performed before and after treatment, and the disappearance of gasping, antipyretic, disappearance of moist rale in the lungs, and absorption time of chest X-ray were observed and recorded. the flow cytometry was used to detect peripheral blood levels of CD4+ and CD8+, and serum levels of IL-6 and IL-8 were detected by ELISA. The clinical effect rate was evaluated.@*Results@#The total effective rate was 97.5% (118/121) in the observation group and 91.7% (110/120) in the control group,with statistically significant differences (χ2=4.406, P=0.044). After treatment, the scores of cough, phlegm, shortness of breath, pale face, fatigue, spontaneous perspiration, poor appetite in the observation group were significantly lower than those in the control group (t value were 10.008, 11.529, 11.778, 12.043, 11.309, 10.128, 11.008, P<0.01), and elapsed time of gasping, antifebrile time, disappear time of lung moist crackles, absorption time were significantly earlier than the control group (t value were 15.121, 10.099, 18.441, 14.310, P<0.01). After 2 week’ treatment, the scores of symptoms of deficiency of lung-spleen Qi in the observation group were significantly lower than those of control group (P<0.01). Compared with the control group, the duration of wheezing and antipyretic, duration of lung dampness and chest X-ray absorption in the observation group were significantly decreased (P<0.05). The total effective rate of the observation group was 97.52%, which was significantly higher than the control group 92.67% (P<0.05). After 2 week treatment, the CD4+ (41.03% ± 4.55% vs. 33.52% ± 3.95%, t=14.011) in the observation group was significantly higher, CD8+ (22.71% ± 2.83% vs. 26.65% ± 3.05%, t=10.061) was significantly lower than the control group, serum levels of IL-6 (28.13 ± 3.63 ng/L vs. 40.09 ± 4.61 ng/L, t=14.890) and IL-8 (21.41 ± 2.75 ng/L vs. 29.05 ± 3.61 ng/L, t=11.439) were significantly lower than the control group (P<0.01).@*Conclusions@#Massage combined with microwave therapy for the Children's bronchial pneumonia with deficiency of lung-spleen Qi syndrome can promote the improvement of symptoms, enhance the efficacy, increase the immune function and reduce inflammation level.
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Objective To evaluate the efficacy and safety of the mobilization and collection of unrelated allogeneic peripheral blood stem cells. Methods The suitable stem cell mobilization plan was made in accordance with the hematopoietic stem cell mobilization plan of China Marrow Donor Program, the ruler of the hospital, and the donor's constitution. The unrelated allogeneic peripheral blood stem cells of 64 healthy donors were collected in the second Hospital of Shanxi Medical University from May 2012 to January 2017. The donor was infected one or several times with the mobilization agent granulocyte colony stimulating factor (G-CSF) by 5-10 μg·kg-1·d-1. After 3-4 days, peripheral blood hematopoietic stem cells were collected using COBE Spectra blood cell separator. Then, the effect and adverse reaction of donors were analyzed from different age and sex. Results It can achieve the acquisition requirements using 3 or 4 days of mobilization programs, mononuclear cells≥5.0×108/kg, CD34+cells≥2.0×106/kg. The single acquisition success rate (the target acquisition of the number of mononuclear cells and CD34+) up to 65 ％, collection efficiency reached 52％, which could reduce the risk of donor and the cost of patients. The quality of donor stem cell of young was better than that of older persons. Sixteen donors (25％) had mild adverse reactions, and no special treatment was required. Conclusions Allogeneic stem cell mobilization is safe. Starting from save medical resources and the interests of the donor the 3 day or 4 days of mobilization scheme could improve the success rate of the single mobilization. During the collection process, the condition of donor hypocalcemia should be observed and health education should be given to relieve the tension of donor.
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In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.
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In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni+-nitrilotriacetic acid (Ni+-NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni+-NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.
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In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.
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A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent signal in the presence of genotoxic agents, such as Mitomycin C (MC). The performance of this whole-cell optical fiber sensor system was examined as a function of several parameters, including gel probe thickness, bacterial cell density, and diameter of the fiber-optic core and working temperature. An optimal response to a model genotoxicant, Mitomycin C, was achieved with agar-bacterial gel membrane: the thickness of gel membrane was about 5 mm; the cell density of bacteria in gel membrane was about 2.0 x 10(7)/ml; the diameter of fiber-optic core was 5.0 mm; the working temperature was 25 degrees C. Under these optimized conditions, the response time was less than 10 h to Mitomycin C, with a lower detection threshold of 0.1 mg/L.
Subject(s)
Biosensing Techniques , Fiber Optic Technology , Genetic Variation , Luminescent Measurements , Luminescent Proteins , Genetics , Mitomycin , Pharmacology , Toxicity , Optical Fibers , Photobacterium , Genetics , Transcription, GeneticABSTRACT
A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent signal in the presence of genotoxic agents, such as Mitomycin C (MC). The performance of this whole-cell optical fiber sensor system was examined as a function of several parameters, including gel probe thickness, bacterial cell density, and diameter of the fiber-optic core and working temperature. An optimal response to a model genotoxicant, Mitomycin C, was achieved with agar-bacterial gel membrane: the thickness of gel membrane was about 5 mm; the cell density of bacteria in gel membrane was about 2.0 x 10(7)/ml; the diameter of fiber-optic core was 5.0 mm; the working temperature was 25 degrees C. Under these optimized conditions, the response time was less than 10 h to Mitomycin C, with a lower detection threshold of 0.1 mg/L.
Subject(s)
Biosensing Techniques , Luminescent Measurements , Fiber Optic Technology , Luminescent Proteins/genetics , Mitomycin/pharmacology , Mitomycin/toxicity , Photobacterium/genetics , Transcription, Genetic/drug effects , Genetic VariationABSTRACT
The luminous intensity of dark variant (S1) separated from photobacterium phosphoreum (A2) was 1/10,000 less than that of wild-type. Ethidium bromide (EB) (0.6 mg/L), Mytomycin C (MC, 0.05 mg/L), 2-amino fluorene (2-AF, 1.0 mg/L) all could strongly induce reversion mutation for S1 within 24 h and increase reversion ratio significantly. The results of experiments indicated that these revertants had stable genetic characteristic and the mutation may take place at gene levels. The mutagenesis to S1 caused by EB, MC and 2-AF was detected and it may be used as a new rapid, simple and sensitive method for gene toxicant monitoring.
Subject(s)
Ethidium , Pharmacology , Toxicity , Genetic Variation , Luciferases , Luminescent Measurements , Mitomycins , Pharmacology , Toxicity , Mutagens , Mutation , Photobacterium , Genetics , Toxicology , Methods , Transcription, GeneticABSTRACT
The luminous intensity of dark variant (S1) separated from photobacterium phosphoreum (A2) was 1/10,000 less than that of wild-type. Ethidium bromide (EB) (0.6 mg/L), Mytomycin C (MC, 0.05 mg/L), 2-amino fluorene (2-AF, 1.0 mg/L) all could strongly induce reversion mutation for S1 within 24 h and increase reversion ratio significantly. The results of experiments indicated that these revertants had stable genetic characteristic and the mutation may take place at gene levels. The mutagenesis to S1 caused by EB, MC and 2-AF was detected and it may be used as a new rapid, simple and sensitive method for gene toxicant monitoring.