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OBJECTIVE@#To investigate the expression and localization of metabotropic glutamate receptors 7 and 8 (mGluR7/8) in rat superior cervical ganglion (SCG) and their changes in response to chronic intermittent hypoxia (CIH).@*METHODS@#We detected the expressions of mGluR7 and mGluR8 in the SCG of 8-week-old male SD rats using immunohistochemistry and characterized their distribution with immunofluorescence staining. The expression of mGluR7 and mGluR8 in the cytoplasm and nucleus was detected using Western blotting. A 6-week CIH rat model was established by exposure to intermittent hypoxia (6% oxygen for 30 s followed by normoxia for 4 min) for 8 h daily, and the changes in systolic blood pressure, diastolic blood pressure and mean arterial pressure were measured. The effect of CIH on expression levels of mGluR7 and mGluR8 in the SCG was analyzed using Western blotting.@*RESULTS@#Positive expressions of mGluR7 and mGluR8 were detected in rat SCG. mGluR7 was distributed in the neurons and small fluorescent (SIF) cells with positive staining in both the cytoplasm and nuclei, but not expressed in satellite glial cells (SGCs), nerve fibers or blood vessels; mGluR8 was localized in the cytoplasm of neurons and SIF cells, but not expressed in SGCs, nerve fibers, or blood vessels. Western blotting of the nuclear and cytoplasmic fractions of rat SCG further confirmed that mGluR7 was expressed in both the cytoplasm and the nucleus, while mGluR8 exists only in the cytoplasm. Exposure to CIH significantly increased systolic blood pressure, diastolic blood pressure and mean arterial pressure of the rats (all P < 0.001) and augmented the protein expressions of mGluR7 and mGluR8 in the SCG (P < 0.05).@*CONCLUSION@#mGluR7 and mGluR8 are present in rat SCG but with different localization patterns. CIH increases blood pressure of rats and enhanced protein expressions of mGluR7 and mGluR8 in rat SCG.
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Male , Animals , Rats , Rats, Sprague-Dawley , Superior Cervical Ganglion , Receptors, Metabotropic Glutamate , HypoxiaABSTRACT
Objective:To investigate the effect of different incubation time of aminolevulinic acid (ALA) on photodynamic inhibition of Propionibacterium acnes biofilms. Methods:Propionibacterium acnes biofilms were formed in 24-well plates with pre-placed cell slides and 96-well plates. The formation of the biofilm structure was observed by confocal laser scanning microscopy (CLSM) , and the growth activity of the biofilm was assessed by the tetrazolium salt XTT assay. The in vitro successfully constructed biofilm models were divided into 6 groups: negative control group receiving neither ALA treatment nor LED radiation, ALA group incubated with ALA alone for 30 minutes, LED group receiving LED radiation alone, ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group incubated with ALA for 15, 30 and 60 minutes respectively followed by LED radiation. After the treatment, CLSM was performed to observe the biofilm structure, as well as to determine the dead/living bacteria ratio, and XTT assay to assess the growth activity of the biofilm. Differences among groups were analyzed using one-way analysis of variance and least significant difference- t test. Results:CLSM showed that the Propionibacterium acnes biofilm model was successfully constructed in vitro. The dead/living bacteria ratios were 0.90 ± 0.16, 1.75 ± 0.19, and 2.57 ± 0.32 in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group respectively, which were significantly higher than the dead/living bacteria ratio in the negative control group (0.31 ± 0.01; t= 55.56, 138.62, 74.64, respectively, all P<0.001) ; the biofilm viability value was significantly lower in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group (0.35 ± 0.02, 0.26 ± 0.02, 0.18 ± 0.01, respectively) than in the negative control group (0.43 ± 0.00; t= 35.66, 2.64, 110.96, respectively, all P < 0.001) . CLSM showed that the structure of the Propionibacterium acnes biofilm was destroyed under the action of ALA-PDT, and the destruction was aggravated with the prolongation of incubation time of ALA. Conclusion:The prolongation of incubation time of ALA can enhance the inhibitory effect of ALA-PDT on Propionibacterium acnes biofilms.
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Objective:To analyze the molecular epidemiological characteristics of the Corona virus disease 2019 (COVID-19) cases in Shijiazhuang, which can reveal the origin of the outbreak and provide a scientific basis for COVID-19 prevention and control.Methods:From January 2 to January 8, 2021, a total of 404 samples from 170 COVID-19 cases were collected from the Shijiazhuang Fifth Hospital. The consensus sequence of 2019 novel Coronavirus(2019-nCoV) was obtained through multiplex polymerase chain reaction-based sequencing. The sequences of 170 COVID-19 cases were analyzed by the PANGOLIN, and the data were statistically analyzed by T-test.Results:Among the 404 COVID-19 samples, a total of 356 samples obtained high quality genome sequences (>95%,100×sequencing depth). The whole genome sequences of 170 COVID-19 cases were obtained by eliminating repeated samples. All 170 sequences were recognized as lineage B1.1 using PANGOLIN. The number of single nucleotide polymorphism arrange from 18-22 and most of the single nucleotide polymorphism were synonymous variants. All of 170 genomes could be classified into 48 sub-groups and most of the genomes were classified into 2 sub-groups (66 and 31, respectively).Conclusions:All cases in this study are likely originated from one imported case. The viruses have spread in the community for a long time and have mutated during the community transmission.
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Jianwei Xiaoshi tablets， as a common variety of Chinese patent medicine with "one product with many manufacturers"， have many manufacturers and huge market sales. However， the phenomenon about uneven quality and discrepant price is prominent. Based on this， this study was carried out for the quality evaluation of Jianwei Xiaoshi tablets by applying the high-quality evaluation criteria with the quality as core for Chinese patent medicine， which was based on the full production cycle， from the multi-dimension including raw material selection， production process， quality control， post-marketing research and so on. The evaluation results showed that the quality evaluation scores of Jianwei Xiaoshi tablets from different manufacturers varied greatly （ranging from 35 to 66）， indicating that the quality was significantly different. In the actual production， generally inadequate attention was paid to the quality of raw materials， and the quality of raw materials was insufficient with the score ratio of 43%， especially the poor consistency control of them. The role of good manufacturing practice was obvious， and the scores of production process were generally high with the average score ratio of 62%， and the maximum up to 80%. The technological advancement of the manufacturer was outstanding. The score ratio of quality control was only 31% that the internal quality standard of each manufacture almost stayed at the qualified line， which was equal to the national standard， and the consistency of products was insufficient. The post-marketing research was lacking with the score ratio of 37%. Manufacturers with high brand awareness and market share were upper scores， while the others lagged far behind. The results of this evaluation are in line with the overall prediction， which can provide a reference for the high-quality evaluation of Chinese patent medicine， and supply the scientific data for high-quality and high-price application.
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The blood donation, component preparation and manufacturing, laboratory, issuing and quality control were studied and compared between the UK and China, in order to learn British experience in the clinical practice and blood services. The age limits of blood donors, blood collection units, donation times per year, laboratory items, and the types(volumes) of component preparation and manufacturing in the UK are more superior than those in China. In addition, the blood quality monitoring and regarding indicators are more scientific and reasonable in the UK. The full reimbursement of clinical blood expenses for patients has been realized in the UK. The British experience in continuous safeguard of the blood safety and balance of requirement and availability contributes to the constant and scientific development of British blood services over the years, and is worthy of references.
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To investigate the effect of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on () biofilm. biofilms were constructed on a cell slide and treated with ALA-PDT.According to different light doses,the biofilms were divided into six groups:ALA-PDT group [ALA-PDT1 (50 J/cm),ALA-PDT2 group (100 J/cm),ALA-PDT3 group (200 J/cm)],ALA-only group (ALA group),light-only group (LED),and a negative control group (ALA-PDT-group).The biofilm structure and the ratio of the dead bacteria/live bacteria were observed using a laser confocal microscope (CLSM).Biofilm viability was measured using the XTT assay. CLSM showed that the biofilm structures of ALA group and LED group were not significantly different from that of ALA-PDT-group,whereas the biofilm structure was more seriously damaged in ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group than in the ALA-PDT-group.The ratios of the dead/live bacteria in ALA-PDT-group,ALA group,LED group,ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group were 0.350±0.033, 0.305±0.046, 0.330±0.032, 1.525±0.439, 2.293±0.148 and 3.092±0.189,respectively.ALA group(=0.003, =1.000)and LED group(=-0.025, =1.000)did not significantly differ from the ALA-PDT-group.However,the ratio of dead/live bacteria in ALA-PDT-group was significantly lower than those in ALA-PDT1 group (=-0.162, <0.001),ALA-PDT2 group (=-0.254, <0.001),and ALA-PDT3 group (=-0.352, <0.001).The values of the XTT assay were were 0.462±0.028,0.465±0.044,0.437±0.047,0.301±0.040,0.207±0.001,and 0.110±0.007,respectively,in ALA-PDT-group,ALA group,LED group,ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group.Although the values of XTT assay in ALA(=-0.044, =1.000)and LED groups (=-0.020, =1.000)did not significantly differ from that in ALA-PDT-group,it was significantly higher in ALA-PDT-group than in ALA-PDT1 group (=1.175, <0.001),ALA-PDT2 group (=1.942, <0.001),and ALA-PDT3 group (=-0.352, =2.742, <0.001). ALA-PDT has an inhibitory effect on biofilm.ALA-PDT destroys biofilm structure and inhibits biofilm viability.
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Aminolevulinic Acid , Biofilms , Photochemotherapy , Photosensitizing Agents , Propionibacterium acnesABSTRACT
Skin microbiota begin to form shortly after birth,with a wide variety and huge quantity,and are closely related to skin health.Acne is a common skin disorder associated with microbial infections.Propionibacterium acnes,Staphylococcus,Malassezia,etc.,are three species of microorganisms currently known to be most closely associated with the occurrence of acne.This review summarizes the relationship between the above three species of microorganisms and the occurrence of acne,and mainly includes the pathogenicity of bacteria or fungi of different types,the relationship between these bacteria or fungi and the pathogenesis of acne,the difference in host immune responses induced by these bacteria or fungi,so as to provide evidence for developing new strategies for the treatment of acne.
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Objective@#To explore the degree of social support in patients after Coronary Artery Bypass Grafting(CABG) and its influence on Acute Stress Disorder (ASD), in order to provide theory evidence for nursing intervention.@*Methods@#From June 2017 to Mar 2018, 359 patients after CABG in Tangshan Workers Hospital were recruited for the study. Their general information were collected, SASRQ, SSRS and PSS-Fa were used for professional evaluation.@*Results@#The degree of social support in patients after CABG was at medium level, the total score of social support was (41.69±0.28), higher than the normal domestic value[(34.56±3.73)score, n=128]; character (F=29.652), employment (t=6.526)and the type of health insurance (F=20.547)impacted on the degree of social support (P<0.05). Total score of social support had negative correlation with SASRQ total score (r=-0.528, P<0.05); character (β=0.262), family support (β=-0.281) and social support (β=-0.267) were picked in the regression equation (P<0.05), the results showed that introversion was a risk factor for ASD in patients after CABG, high degree of family support and social support were protective factors for ASD in patients after CABG.@*Conclusions@#Social support of patients after CABG needs to be improved, so as to reduce the incidence of acute stress disorder.
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Objective To explore the degree of social support in patients after Coronary Artery Bypass Grafting(CABG) and its influence on Acute Stress Disorder (ASD), in order to provide theory evidence for nursing intervention. Methods From June 2017 to Mar 2018, 359 patients after CABG in Tangshan Workers Hospital were recruited for the study. Their general information were collected, SASRQ, SSRS and PSS-Fa were used for professional evaluation. Results The degree of social support in patients after CABG was at medium level, the total score of social support was (41.69±0.28), higher than the normal domestic value[(34.56 ± 3.73)score, n=128]; character (F=29.652), employment (t=6.526)and the type of health insurance (F=20.547)impacted on the degree of social support (P＜0.05). Total score of social support had negative correlation with SASRQ total score (r=-0.528, P＜0.05); character (β=0.262), family support (β=-0.281) and social support (β=-0.267) were picked in the regression equation (P＜0.05), the results showed that introversion was a risk factor for ASD in patients after CABG, high degree of family support and social support were protective factors for ASD in patients after CABG. Conclusions Social support of patients after CABG needs to be improved, so as to reduce the incidence of acute stress disorder.
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Objective To investigate the effect of conbercept on rabbit's haze after photorefractive keratectomy (PRK).Methods Sixty-four pedigree New Zealand white rabbits were randomly divided into PRK group,normal saline solution group,conbercept 0.5 mg group,conbercept 1.0 mg group,with 16 rabbits in each group.PRK was performed on right eyes,PRK group only received PRK,the other three surgery groups were given postoperative subconjunctival injection of 0.05 ml normal saline solution,0.5 mg (0.0:5 ml) conbercept and 1.0 mg(0.10 ml) conbercept,respectively.In addition,another 8 rabbits were randomly chosen as normal control group.The healing of postoperative corneal epithelial was observed by slit lamp biomicroscope,and the degrees of haze were graded based on Fantes.Eight rabbits in the surgery groups and 4 rabbits in the normal control group were killed in the first week and the fourth week.The corneal tissue was stained by hematoxylin-eosin,and the expressions of transforming growth factor-β1 (TGF-β1),α-smooth muscle actin (α-SMA) and matrix metalloproteinase-2 (MMP-2)were detected by immunochemistry.The use and feeding of experimental animals followed the Relevant Regulations of the Animal Management Committee of Binzhou Medical University.This study protocol was approved by Ethic Committee of the Affiliated Hospital of Binzhou Medical University (No.201701-08).Results Corneal epithelium of all operative rabbits healed completely at 3-5 days and no significant difference in healing time between the groups after operation (F=0.37,P =0.77).The degree of haze in each surgery group reached the highest value at about 4 weeks after operation,and haze in the conbercept 1.0 mg group was the most serious,followed by PRK group and normal saline solution group,haze in conbercept 0.5 mg group was significantly alleviated (Fgroup =20.114,P =0.000;Ftime =8.084,P =0.006).Hematoxylin-eosin staining showed that corneal epithelial cells and fibroblasts in superficial stroma proliferated in one week after PRK,which lead to the disorder of cells and collagen fibers,and the extent of hyperplasia was the same as that of haze.Immunohistochemistry showed that at one week after operation,the expressions of factors in PRK group and normal saline solution group were apparently lower than that of conbercept 1.0 mg group,but were apparently higher than that of conbercept 0.5 mg group,and the expressions of the factors were the weakest in normal control group,with significant differences between them (all at P ＜ 0.05).Conclusions Subconjunctival injection of appropriate conbercept can inhibit the formation of haze and the proliferation of corneal epithelium and superficial stroma,but overdose of conbercept leads to opposite effects.
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Objective To evaluate the effect of Aspergillus fumigatus on the expression of tumor necrosis factor-α (TNF-oα) and activation of intracellular signaling molecule p38 mitogen-activated protein kinase (p38MAPK) in a human acute monocytic leukemia cell line THP-1.Methods Cultured THP-1 cells (2 x 105/ml) were divided into 4 groups to be treated with Aspergillus fumigatus suspensions at concentrations of 106 and 107 colony-forming units (CFU)/ml (106-and 107-CFU/ml Aspergillusfumigatus groups),100 mg/L β-glucan (a positive stimulus,β-glucan group),culture medium (blank control group) respectively for 1,3 and 6 hours.Real-time fluorescence-based quantitative PCR (qPCR) was conducted to determine the mRNA expression of TNF-α in the THP-1 cells in the above groups.Some other THP-1 cells were treated with 107 CFU/ml Aspergillusfumigatus suspensions (107-CFU/ml Aspergillusfumigatus group),β-glucan (β-glucan group) and culture medium (blank control group) separately for 24 hours,and enzymelinked immunosorbent assay (ELISA) was performed to detect the level of TNF-α in the culture supernatant of THP-1 cells.Western blot analysis was conducted to detect the levels of p38MAPK and phosphorylated p38MAPK in THP-1 cells after 15-,30-and 60-minute treatment with 107 CFU/ml Aspergillusfumigatus suspensions.After 2-hour incubation with the p38MAPK inhibitor SB203580 (20 μmol/L),some THP-1 cells were additionally treated with 107 CFU/ml Aspergillus fumigatus suspensions,β-glucan and culture medium separately for 6 hours,and those without SB203580 treatment served as the control group.Then,qPCR was performed to measure the mRNA expression of TNF-α in the THP-1 cells in the above groups.Results The mRNA expression of TNF-α significantly differed among the 106-and 107-CFU/ml Aspergillus fumigatus groups,β-glucan group and blank control group (F =110.983,P ＜ 0.001),and significantly increased over time (F =701.680,P ＜ 0.001).After 24-hour treatment with 107 CFU/ml Aspergillus fumigatus suspensions,the TNF-α level(6 236.30 ± 437.12 ng/L)significantly increased compared with the blank control group (132.10 ± 0.61 ng/L,P ＜ 0.01).Thirty minutes after the treatment with 107 CFU/ml Aspergillusfumigatus suspensions,the phosphorylated p38MAPK level significantly increased,but started to decrease at 60 minutes.The mRNA expression of TNF-α was significantly lower in the SB203580-treated Aspergillusfumigatus groups (3.83 ± 0.62) than in the SB203580-untreated Aspergillus fumigatus groups (187.23 ± 21.62).Conclusion After the treatment with Aspergillus fumigatus,human THP-1 cells can activate the signal molecule p38MAPK and secrete TNF-α,suggesting that monocytes may participate in the innate immune response to Aspergillusfumigatus infection.
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Objective To determine the expression of interleukin-6 (IL-6) in cystic lesions of patients with acne vulgaris,and to evaluate the in vitro effect of Propionibacterium acnes (P.acnes) on the production of IL-6 and activation of p38 mitogen-activated protein kinase (p38MAPK) in the human acute monocytic leukemia cell line THP-1.Methods Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of IL-6 in cystic lesions of 6 patients with acne vulgaris,as well as in skin tissues of 6 healthy persons.Some cultured THP-1 cells were divided into 5 groups to be treated with 2 × 106 CFU/ml,2 × 107 CFU/ml and 2 × 108 CFU/ml heat-killed P.acnes suspensions (P.acnes groups),100 μμtg/L lipopolysaccharide (LPS group) and RPMI 1640 medium (control group) respectively.After 1-,3-and 6-hour treatment,real-time fluorescence-based quantitative PCR was conducted to determine the mRNA expression of IL-6 in the above groups.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of IL-6 in the culture supernatant of cells in the 2 × 108-CFU/ml P.acnes group,LPS group and control group at 24 hours after the treatment.Western blot analysis was conducted to determine the protein expression of p38MAPK and phosphorylated p38MAPK in the 2 × 108-CFU/ml P.acnes group after 15-,30-and 60-minute treatment,as well as in the LPS group after 30-minute treatment and in the control group.Some other THP-1 cells were divided into 3 groups:2 × 108-CFU/ml P.acnes group treated with 2 × 108 CFU/ml P.acnes suspensions,SB203580 (an inhibitor of p38MAPK) group treated with 20 μmol/L SB203580 for 30 minutes followed by the treatment with 2 × 108 CFU/ml P.acnes suspensions,and control group treated with RPMI 1640 medium alone.After 6-hour treatment,the mRNA expression of IL-6 in the above 3 groups was measured by real-time fluorescencebased quantitative PCR.Results The mRNA expression of IL-6 was significantly higher in the cystic lesions of acne vulgaris than in the normal skin tissues (3.680:±:0.790 vs.1.155 ± 0.250,t =3.047,P ＜0.05).Two-way analysis of variance showed that there were significant difference in the mRNA expression of IL-6 among the 2 × 106-CFU/ml,2 × 107-CFU/ml and 2 × 108-CFU/ml p.acnes groups,LPS group and control group (F =532.3,P ＜ 0.001,v =4),and the mRNA expression of IL-6 significantly differed among different time points (F =526.6,P ＜ 0.001,v =2).There were also significant differences in the IL-6 level in the culture supernatant of cells among the 2 × 108-CFU/ml p.acnes group ([1 618.22 ± 32.23] ng/L),LPS group ([3 212.06 ± 353.00] ng/L) and control group ([147.10 ± 0.53] ng/L;v =2,F =102.35,P ＜0.01).After 15-,30-and 60-minute treatment with 2 × 108 CFU/ml P.acnes suspensions,the protein expression of phosphorylated p38MAPK obviously increased.The mRNA expression of IL-6 in THP-1 cells was significantly lower in the SB203580 group than in the 2 × 108-CFU/ml p.acnes group (t =15.91,P =0.004).Conclusions The mRNA expression of IL-6 evidently increases in the cystic lesions of patients with acne vulgaris.P.acnes can activate the signaling molecule p38MAPK in THP-1 cells,and promote the production of IL-6 by THP-1 cells.
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Objective To construct a native promoter-regulated Aspergillus fumigatus strain containing red fluorescent protein-labeled calmodulin (CaM-RFP),and to observe the dynamic distribution of calmodulin during the growth of Aspergillus fumigatus.Methods Bilateral flanking sequences of Aspergillus fumigatus calmodulin gene were designed,and plasmids containing the two flanking sequences and mRFP-Aspergillus fumigatus pyrG gene (mRFP-AfpyrG) were amplified separately.The final linear PCR product for transformation was generated from the above three PCR products by fusion PCR.Then,the above linear fragment was transferred into the Aspergillus fumigatus strain by protoplast transformation,so as to construct the CaM-RFP Aspergillus fumigatus strain.The monoclonal colony was picked from the screening medium and subjected to culture.Then,the stablest fluorescent monoxenic strain of Aspergillus fumigatus was selected,and the transformant was verified by PCR.The recombinant strain and wild-type stain were cultured on solid nutrient media separately,and the morphology of these strains was observed by fluorescence microscopy at different time points.Additionally,the above 2 strains were cultured in liquid media separately,and XTT assay was performed to evaluate the growth activity of strains.Microscopy was also conducted to dynamically observe the CaM-RFP Aspergillusfumigatus strain,and analyze the spatial and temporal distribution of calmodulin during the growth and development of Aspergillus fumigatus.Results The fluorescent phenotype and PCR identification results both indicated the successful construction of the CaM-RFP Aspergillus fumigatus strain.The growth activity at 24 hours did not differ between the recombinant strain and wild-type stain (A490:0.689 ± 0.081 vs.0.678 ± 0.054,t =1.32,P ＞0.05),so did the morphology.During the polarized growth of Aspergillus fumigatus,calmodulin was always at the top of the hyphae,germination site of the hyphal branch and the top of new branches.Conclusion Calmodulin may be involved in the regulation of spore germination and polar hyphal growth of Aspergillus fumigatus.
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Background Pigmentary glaucoma and pseudoexfoliation glaucoma are characterized by pigment dispersion in trabecular meshwork,and the dipersitional pigment probably contributes to the resistance of aqueous outflow pathway,irreversible damage of the trabecular meshwork and the remodeling abnormality of extracellular matrix.Researches determined that contents of transforming growth factor-β (TGF-β) in the aqueous humor are increased in glaucomatous eyes.However,the effects of TGF-β and fibronectin (FN) in the trabecular meshwork cells (TMCs) acted by iris pigmentary particles still are not elucidated.Objective This study was to investigate the effects of iris pigment particles on TGF-β1 and FN expression in bovine TMCs (BTMCs) cultured in vitro.Methods BTMCs were cultured in vitro by explant culture method and identified by morphological evaluation.The third generation of BTMCs were divided into normal control group and pigment group,and 100 μ1 PBS and 100 μl iris pigment suspension (final concentration of 1 × 107 particles/ml) were added into the medium for 24 hours,respectively.The expressions of TGF-β1 mRNA,FN mRNA and their proteins in the BTMCs were assayed by real-time fluorescence quantitative PCR and ELISA,respectively.Results Cultured cells grew well and showed the fusiform,polygon and dendritic like in shape,with pigmented and round nuclei.The relative expression levels of TGF-β1 mRNA and FN mRNA in the cells were 2.98±0.27 and 0.36±0.10 in the iris pigment group,which were significantly higher than 1.00±0.00 and 1.00±0.00 in the normal control group (t =12.68,10.60,both at P =0.00).The concentrations of TGF-β1 protein and FN protein in the cell suspension were (156.60±9.74)ng/L and (59.29±15.79)mg/ml in the iris pigment group,showing significant differences in comparison with (65.46 ± 14.24) ng/L and (102.10 ± 12.14)mg/mlin the normal control group (t=9.15,P=0.00;t=3.72,P=0.02).Conclusions The expression of TGF-β1 is up-regulated and that of FN is down-regulated in BTMCs cultured by iris pigment,inferring that TGF-β1 and FN participate in the pathogenesis and development of pigmentary and pseudoexfoliation glaucoma.
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Background Recent studies have confirmed that sulforaphane (SFN) can activate multiple pathways,and promote the expression of the antioxidants in cells.Thioredoxin (Trx) plays an important role in maintaining the intracellular redox in the steady state.Objective This study was to investigate the effect and mechanism of SFN on Trx expression in bovine trabecular meshwork cells (BTMCs) cultured in vitro.Methods BTMCs were cultured in vitro and identified by morphological evaluation.The third generation of BTMCs were cultured in the medium with 0,10,20 and 30 μmol/L SFN for 30 minutes.Real-time PCR was applied to measure the expression of Trx mRNA in BTMCs.The BTMCs were randomly divided into normal control group,LY294002 group,U0126 group,SFN group,LY294002 +SFN group and U0126+SFN group.The expressions of Nrf2 protein and Trx protein in each group were measured by Western blot.Results The BTMCs was successfully cultured in vitro.The expressions of Trx mRNA were significantly different among the different concentrationss of SFN treatment (F=88.090,P＜0.01).The expressions of Trx protein and Nrf2 protein in the LY294002 +SFN group,U0126 +SFN group and SFN group were significantly higher than those in the normal control group (all at P ＜ 0.01).The expressions of Trx protein and Nrf2 protein in the LY294002+SFN group and U0126+SFN group were significantly higher than those in the SFN group (all at P＜0.01).Conelusions SFN can activate Nrf2 by phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases 1/2 (ERK1/2) signaling pathways,which can increase the expression level of Trx in BTMCs cultured in vitro.
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Background Evidences indicated that oxidative stress damage is an essential pathological process in primary open angle glaucoma.Sulforaphane (SFN) can play an antioxidative stress role to many tissues and cells by activating Nrf2/ARE single pathway.However,whether SFN has a protective role to oxidative stress induced damage of trabecular meshwork cells is still unclear.Objective This study was to investigate the antioxidant effect of SFN against H2O2-induced oxidative damage in bovine trabecular meshwork cells.Methods Trabecular cells were isolated from fresh black bovine eyeballs and primarily cultured and passaged.The third generation of cells were incubated to 96-well dish at a density of 1 ×103/well for 24 hours and divided into 4 groups.The cells were incubated using 100 μl serum-free medium in the blank control group.Oxidative damage models were established by adding 100 μmol/L H2O2(100 μl) in medium in the H2O2 group.The cells were cultured with the medium containing 10 μmol/L SFN (100 μl) in the SFN group,and 100 μl H2O2 at the final concentration of 100 μmol/L was added in the SFN-treated cell medium in the SFN +H2O2 group.The cell vitality in various groups was assayed by using cell counting kit-8 (CCK-8).The apoptosis rate of the cells was detected by Annexin V-FITC/PI double-staining with flow cytometry.Results Cultured cells showed a spindle shape with uniform size,abundant cytoplasm,numberous pigmented particles and big nucleolus.The relative cell vitality reduced to (67.00± 1.27)％ and (80.00±6.25)％ in the H2O2 group and SFN+H2O2 group in comparison with 100％ in the blank control group,and the cell vitality in the SFN+ H2O2 group was lower than that in the SFN group but higher than that in the H2 O2 group (both at P＜0.01).The mean apoptosis rate was (11.33 ±0.77) ％,(32.31 ± 1.03) ％,(10.44 ±0.68) ％ and (17.68 ±0.21) ％ in the blank control group,H2 O2 group,SFN group and SFN+H2O2 group,respectively,showing a significant difference among the groups (F=539.96,P＜0.01),and the apoptosis rate in the SFN+H2O2 group was significantly lower than that in the H2O2 group but higher than that in the blank control group and SFN group (all at P＜0.01).Conclusions SFN can improve the antioxidative stress ability of trabecular meshwork cells and alleviate the damage induced by oxidative stress.
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Objective To investigate expressions of surface maturation markers, secreting cytokines and the stimulat?ing effect on T lymphocytes in the dendritic cells (DC) from peripheral blood of tuberculosis (TB) patients after loading dose treatment. Methods TB patients who received initial treatment (n=68) were collected at the fifth hospital of Shijiazhuang from 2013 June to 2014 January. Base on clinical diagnosis and treatment guidelines of tuberculosis, they were divided into sputum smear-negative group (35 cases) and sputum smear-positive group(33 cases). Forty cases of healthy adult were se?lected as control group. Mononuclear cells were isolated from peripheral blood and were cultured in medium to differentiate into DCs. Expression levels of CD83 and CD86 on DCs were examined by flow cytometry. The proliferation of allogeneic mixed lymphocyte stimulated by DCs was dectected using MTT assay. Contents of IL-12, IL-10 and INF-γin the cultural supernatant of DCs and blood serum from TB patients were detected by ELISA. Results Compared with controls ,the ex?pressions of CD83 and CD86 on DCs in TB patients after loading dose treatment decreased obviously(P0.05). Conclu?sion The expressions of maturation markers of DC cells of the peripheral blood in TB patients after initial treatment de?creased. The ability of stimulating mixed lymphocyte proliferation is also significantly reduced while secretion of IL-12 was enhanced.
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Objective To develop an intelligent moxibustion instrument based on MCU,on the basis of moxibustion principles of traditional Chinese medicine.Methods A mixed-signal system-on-chip MCU C8051F020 was used to design four kinds of moxibustion heads to heat,light,and to upload detected temperature to the host microcontroller.Sixteen output ports were designed of which temperature and moxibustion time can be individually set.Results The temperature range of moxibustion head was 30-60℃,with error ≤±3 ℃.The timing range was 10-90 min,and full-scale timing error was ≤±1 min.The average infrared all normal emission rate of moxibustion head was 0.89.Conclusions The test parameters meet the requirements for moxibustion instrument,and the instrument is safe,reliable and simple to operate.
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Objective To evaluate the clinical efficacy and safety of a fractional bipolar radiofrequency device for the treatment of wrinkles of the face and neck.Methods This study enrolled 39 volunteers (including 37 females and 2 males aged 35-60 years) with Fitzpatrick skin type Ⅳ-Ⅴ and Fitzpatrick wrinkle scale score of 4-6.All the subjects received three sessions of treatment with a fractional bipolar radiofrequency device at intervals of 4-6 weeks.Follow-up visits were scheduled at one month after each treatment session for evaluation of efficacy (using standardized photography),subjective satisfaction and adverse effects.Results After three sessions of treatment,all the subjects experienced a significant decrease in Fitzpatrick wrinkle scale compared with baseline values,and wrinkle improvement score increased with the increase in treatment sessions.The Fitzpatrick wrinkle scale score for forehead wrinkles,glabella wrinkles,fishtail lines and neck wrinkles in these subjects after three sessions of treatment significandy differed from that before treatment (all P ＜ 0.01).Significant differences were also observed in the wrinkle improvement score for forehead wrinkles between these subjects after two sessions and one session of treatment (P ＜ 0.01),and observed in that for glabella wrinkles,fishtail lines and neck wrinkles between these subjects after three sessions and one session of treatment (P ＜ 0.05 or 0.01).Side effects were mild with no significant downtime.Conclusion Fractional bipolar radiofrequency may be a safe and effective option for the treatment of wrinkles of the face and neck in individuals with Fitzpatrick skin type Ⅳ-Ⅴ.
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Objective To systematically review the efficacy and safety of acupuncture and moxibustion at Fenglong point (ST40) for the treatment of hyperlipemia.Methods Systematic searches were conducted in PubMed, EMbase, the Cochrane Library, CNKI, VIP, CBM and WanFang Data, assisted by manual retrieval, and the RCTs of comparative study on acupuncture and moxibustion at Fenglong point and oral administration drugs were included. Data were extracted and evaluated by two reviewers independently with a specially-designed extraction form. The Cochrane Collaboration's RevMan 5.0 software was used for Meta analysis.Results A total of 6 RCTs involving 701 patients were included. The results of Meta analysis showed that the total effective rate in acupuncture and moxibustion at Fenglong point for the treatment of hyperlipemia was similar with statins medicine or Xuezhikang capsule. Acupuncture Fenglong point and statins medicine had significant difference in decreasing cholesterol and increasing HDL-C, with less side effects.Conclusion Acupuncture at Fenglong point is safe and effective in the treatment of hyperlipemia, but still needs more high-quality RCTs for confirmation.