2.The protective effects of transfected microRNA-146a on mice with sepsis-induced acute lung injury in vivo
Jianguo ZHANG ; Chengzhi DING ; Qiang SHAO ; Fen LIU ; Zhenguo ZENG ; Cheng NIE ; Kejian QIAN
Chinese Critical Care Medicine 2015;(7):591-594
ObjectiveTo investigate the protective effect of transfected microRNA-146a (miR-146a) on mice with sepsis-induced acute lung injury (ALI) in vivo.Methods Twenty-four healthy male BALB/C mice were randomly divided into sham group, sepsis group, transfection group and transfection control group, eachn = 6. Mice in transfection group were given miR-146a agomir loaded by in vivo-jetPEITM via airway before reproduction of model, and mice in transfection control group were given negative control loaded by in vivo-jetPEITM only via airway. The septic model was reproduced by cecal ligation and puncture (CLP) 12 hours after transfection , and the mice in the sham group underwent laparotomy and closure only without ligation and puncture of the cecum. The mice of each group were sacrificed at 24 hours post-operation. The expression of miR-146a in lung tissue was determined by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the quantity of tumor necrosis factor-α (TNF-α) in the bronchial alveolar lavage fluid (BALF) was determined with enzyme-linked immunosorbent assay (ELISA). The wet/dry ratio of lung (W/D) was determined. The pathohistological changes in the lung were observed and scored. Results The expression of miR-146a showed a significant increase in sepsis group, transfection group and transfection control group, which were (3.56±0.43), (27.64±3.46) and (3.72±0.54) folds of that in sham group, respectively (P< 0.05 orP< 0.01). The miR-146a expression in transfection group was significantly increased compared with sepsis group and transfection control group (bothP< 0.01), but no statistical difference in the expression was found between sepsis group and transfection control group (P> 0.05). Compared with the sham group, higher level of TNF-αin the BALF was found in the sepsis group, transfection group and transfection control group (ng/L: 511.65±43.47, 305.74±34.76, 492.27±42.21 vs. 50.72±7.23, allP< 0.01). The level of TNF-α in transfection group was significantly lower than that in sepsis group and transfection control group (bothP< 0.01). Compared with the sham group, the W/D ratio of lung in sepsis group, transfection group and transfection control group showed a significant increase (6.11±0.32, 5.02±0.29, 6.05±0.43 vs. 4.18±0.10, allP< 0.01). The W/D ratio of lung in transfection group was significantly lower than that of sepsis group and transfection control group (bothP< 0.01). The lung injury score of transfection group was significantly lower than that of sepsis group and transfection control group (6.12±0.75 vs. 10.53±1.52, 9.73±1.08, bothP< 0.01).Conclusions miR-146a agomir loaded by in vivo-jetPEITM instillation into airway was able to increase the expression of miR-146a in the lung tissue of septic mice. Up-regulation of miR-146a inhibit the release of the inflammatory cytokine TNF-α stimulated by sepsis, and alleviate inflammatory reaction and lung tissue injury in mice with sepsis-induced ALI.
3.Anti-inflammatory effect of acetylcholine on lipopolysaccharide induced inflammatory response of alveolar macrophages
Fen LIU ; Ning ZHAO ; Donghai LI ; Zhenguo ZENG ; Qiang SHAO ; Feifei PENG ; Yan WANG ; Kejian QIAN
Chinese Critical Care Medicine 2015;(10):811-815
ObjectiveTo observe the effect of acetylcholine (ACh) on lipopolysaccharide (LPS) induced inflammatory model of rat alveolar macrophages, and to observe the effect of the acetylcholinesterase inhibitor physostigmine (Phy) on the anti-inflammatory effect of ACh.Methods The rat alveolar macrophages NR8383 were cultured in vitro, which were divided into five groups: blank control group, LPS group (stimulated with 1 mg/L LPS for 12 hours), LPS+ ACh group (0.01, 0.1, 1, 10, 100μmol/L of ACh were added for 5 minutes before LPS stimulation), LPS+ Phy group (1 mmol/L Phy was added for 5 minutes before LPS stimulation), and LPS+ ACh+ Phy group (1 mmol/L Phy and 10μmol/L ACh were added for 5 minutes before LPS stimulation). The supernatants were collected in each group, the enzyme-linked immunosorbent assay (ELISA) was used to assay the contents of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, and IL-6). The activity of acetylcholine esterase (AChE ) in the supernatant was also determined.Results① The contents of TNF-α (ng/L: 605.09±57.13 vs. 34.07±8.62), IL-1β (ng/L: 377.09±28.55 vs. 32.33±10.62) and IL-6 (ng/L: 558.04±77.45 vs. 42.62±11.21) in the LPS group were significantly higher than those in the blank control group (allP< 0.05). These results indicated that the inflammatory model of rat alveolar macrophages was constructed successfully.② ACh with the final concentrations of 0.01, 0.1, and 1μmol/L had less influence on the production of TNF-α, IL-1β and IL-6 in the culture supernatants of alveolar macrophages stimulated with LPS compared with LPS group (allP> 0.05). Nevertheless, 10μmol/L and 100μmol/L ACh notably reduced the production of TNF-α (ng/L: 451.19±30.67, 332.19±32.19 vs. 604.96±22.56), IL-1β(ng/L: 261.08±24.78, 143.98±28.39 vs. 367.06±10.44) and IL-6 (ng/L: 342.75±54.60, 235.48±29.75 vs. 562.69±63.34) in the culture supernatants compared with the LPS group (allP< 0.05).③ The activity of AChE in the LPS group was significantly higher than that in the blank control group (kU/L: 5.21±0.63 vs. 3.09±0.10,P< 0.05). The activity of AChE was successfully inhibited by 1 mmol/L acetylcholinesterase inhibitor Phy pretreatment compared with that in the LPS group (1.51±0.12 vs. 5.21±0.63,P< 0.05).④ The level of TNF-α (ng/L: 183.17±35.44 vs. 451.19±30.67), IL-1β (ng/L: 91.49±12.27 vs. 261.08±24.78) and IL-6 (ng/L: 108.17±22.82 vs. 342.75±54.60) in the culture supernatants of LPS+ ACh+ Phy group was significantly decreased as compared with LPS+ ACh group (allP< 0.05).Conclusions ACh with the final concentrations of 10μmol/L and 100μmol/L can inhibit the LPS induced inflammatory reaction in alveolar macrophages. The acetylcholinesterase inhibitor Phy can reinforce the ACh-mediated anti-inflammatory effect on alveolar macrophages inflammatory model.
4.The expression changes of miR-146a in lipopolysaccharide-induced alveolar macrophages
Zhenguo ZENG ; Yong LI ; Fen LIU ; Chengzhi DING ; Qiang SHAO ; Kejian QIAN
Chinese Journal of Emergency Medicine 2011;20(2):134-136
Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P < 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P <0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.
6.Analysis of capabilities of microscopic examinations of Plasmodium in Dongxihu District,Wuhan City
Shukun YU ; Liurong DENG ; Fen SHAO ; Yuxin WANG ; Xiujuan LI ; Qian LIU
Chinese Journal of Schistosomiasis Control 2017;29(1):99-101,104
Objective To investigate and analyze the capabilities of Plasmodium microscopic examinations in medical and health organizations in Dongxihu District,Wuhan City,so as to provide evidences for achieving the standard of malaria elimina?tion and malaria control in the future. Methods The basic information of the laboratorians and the hardware facilities in 15 mi?croscopic examination stations of Plasmodium were collected and analyzed from 2010 to 2015. Meanwhile,the quality of mak?ing,dyeing,cleanliness of all the negatives blood smears were evaluated,and all the positive smears were re?checked in the dis?trict during the period above mentioned. Results Totally,there were 28 laboratorians and 9 qualified microscopes in 15 micros?copy stations in Dongxihu District. The age of the laboratorians in the first?level and third?level organizations was mainly from 40 to 50 years old,but the age of those in the second?level organizations was from 20 to 30 years old. The years for microscopic ex?amination working of most of the laboratorians in the first?level and third?level organizations were more than 15 years,while the working years of those in the second?level organizations were mainly less than 10 years. The education level of the laboratorians in the first?level organization was mainly(50.00%)junior college,while the education levels of those in the second and third or?ganizations were mainly bachelor. From 2010 to 2015,8 561 blood smears were produced and 539 of them were reviewed,the co?incidence rates of review were all 100%,and the average qualified rates of making,dyeing,and cleanliness of the negative blood smears were 82.08%,79.92%and 83.33%,respectively. All the rates mentioned above in 2015 were higher than those in 2010(χ2=26.45,16.85,13.93,all P<0.01). Conclusion The capability of microscopic examinations of Plasmodium has improved obviously in Dongxihu District,but the human resources construction and hardware input in this district should be im?proved,meanwhile,the check on the quality control of microscopic examinations of Plasmodium should be continued,so as to guarantee the sustainable and healthy development of malaria control work in this district.
7.Determination of four quinolones in plasm and urine by using mixed mode solid phase extraction and capillary electrophoresis
Lingguo ZHAO ; Huikai SHAO ; Fen QU ; Wei LI ; Jiaping HE ; Jian CHEN
International Journal of Laboratory Medicine 2015;(16):2329-2331,2334
Objective To develop a new method for the rapid determination of lomefloxacin,gatifloxacin,ciprofloxacin and oflox-acin in plasm and urine by solid phase extraction(SPE)and capillary electrophoresis.Methods The capillary was fused silica capil-lary with id/od of 75/365 μm and effective/total length of 40/47 cm.The running buffer was 40 mmol/L borate buffer at pH 9.0. Separation voltage was 13 kV.Temperature was 20 ℃.Detection wave-length was set at 280 nm.The sample was analyzed after the pretreatment of SPE.Results The analysis of lomefloxacin,gatifloxacin,ciprofloxacin and ofloxacin was completed in 6 minutes with satisfied accuracy and precision.Good linearity was found within the range of 1-40 μg/mL,and the r was 0.998 7,0.997 6, 0.998 3 and 0.994 2 respectively.The recoveries of four quinolones in plasm and urine ranged from 80.1% to 107.6%,and the rel-ative standard deviations(RSD)ranged from 2.1% to 6.2%.Conclusion This method is fast,simple,precise and it might be feasi-ble for the determination of lomefloxacin,gatifloxacin,ciprofloxacin and ofloxacin in plasm and urine samples.
8.Effects of kuntai capsule on quality of life, breast distending pain and vaginal bleeding in women at early stage of menopause.
Jing ZHANG ; Li-Li GONG ; Shao-Fen ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(11):972-976
OBJECTIVETo study the effects of Kuntai Capsule (KTC), a Chinese herbal preparation, on the quality of life (QOL), breast distending pain and vaginal bleeding in women at early stage of menopause.
METHODSA total of 104 women at early stage of menopause, 54 had their uterus existed (Ue) and 50 in-existed (Ui), were enrolled, and they were randomized to the KTC group and the control group, with equal cases of Ue and Ui in each. The KTC group was treated with KTC 4 capsules twice a day; the control group treated with premarin 0.45 mg per day and for those of Ue 2 mg medroxyprogesterone additionally, with the remedies medicated orally for 1 year. All the testees were asked to record everyday their own condition of breast pain and vaginal bleeding and followed-up every 3 months to fulfill the Menopause Specific Quality of Life questionnaire. Ultrasonic examination on pelvis and breast as well as endocrine hormone assays of estradiol (E2) and follicle-stimulating hormone (FSH) were performed before and after the medication term.
RESULTSEffects of treatment in the two groups were different insignificantly in terms of QOL. The women were benefited in vasomotor and physical domains from the 3rd month of medication, and the psychosocial domain was also improved (for Ui initiating from the 3rd month and for Ue from the 6th month). In the domain of sexual life, KTC showed its favorable effect only on Ue beginning from the 9th month, but not on Ui; while all subjects in the control group had their sexual life improved from the 3rd month. In domain of breast pain, the occurrence at various time points between the two groups was insignificantly different, only that the severity in Ue of the control group was more significant from the 1st to 3rd month than that in the KTC group. As for the domain of vaginal bleeding, the uterine membrane was basically normal in both groups either before or after medication, but the incidence and lasting days from the 1st to 3rd month in Ue of the KTC group were significantly lower than those of the control group. Levels of E2 and FSH were not significantly changed after medication in the KTC group, while in the control group, E2 significantly increased and FSH significantly decreased in the women of Ue (P <0.05).
CONCLUSIONKTC could evidently improve the QOL of women at the early stage of menopause, and is of high safety, with less adverse reaction of breast pain and vaginal bleeding, and shows few impact on sexual hormones.
Adult ; Breast ; pathology ; physiology ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; pathology ; Humans ; Menopause ; drug effects ; psychology ; Middle Aged ; Pain ; drug therapy ; pathology ; Quality of Life ; Vaginal Diseases ; drug therapy ; pathology ; psychology
9.Sinonasal primary extramedullary solitary plasmacytoma with Epstein-Barr virus infection: report of a case.
Yan-fen FENG ; Qiu-liang WU ; Yong-sheng ZONG ; Qiong SHAO
Chinese Journal of Pathology 2007;36(10):711-712
Antibodies, Monoclonal
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metabolism
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CD79 Antigens
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metabolism
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Epstein-Barr Virus Infections
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Herpesvirus 4, Human
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isolation & purification
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Humans
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Immunoglobulin G
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metabolism
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Male
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Middle Aged
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Nose Neoplasms
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metabolism
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pathology
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therapy
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virology
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Paranasal Sinus Neoplasms
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metabolism
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pathology
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therapy
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virology
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Plasmacytoma
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metabolism
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pathology
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therapy
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virology
10.Determination of Polybrominated Diphenyl Ethers and Derivatives, Tetrabromobisphenol A, Hexabromocyclododecane in Egg Samples by Gel Permeation Chromatography and Dispersive Solid Phase Extraction
Minjie LI ; Fen JIN ; Lili YANG ; Rina WU ; Hua SHAO ; Maojun JIN ; Jing WANG
Chinese Journal of Analytical Chemistry 2014;(9):1288-1294
A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.