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ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.
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Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.
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Objective To investigate the clinical effect and safety of San' ao Pian's treatment on subacute cough induced by airway inflammation.Methods Ninety cases with subacute cough induced by airway inflammation were selected as our subjects.They were randomly divided into control group(n =45) and San'ao Pian group(n =45).Patients in control group were given regular treatment,and in San'ao Pian group were received San'ao Pian's(1.0 g/time oral,3 times/day) for 2 weeks plus regular treatment.The changes of clinical symptoms such as cough,sputum,and wheeze were observed and recorded at the 3th,7th,14th day.Results (1) The main symptoms regarding of cough were improved in two groups(P ＜ 0.01).The day cough symptoms score in San'ao Pian group were (2.18 ±0.62),(1.22 ±0.46),(0.83 ±0.45) at the 3th,7th,14th days after treatment,lower than that of control group ((2.78 ± 0.55),(2.05 ± 0.41),(1.86 ± 0.68)) and the differences were significant(F within group =10.23,P ＜ 0.05 ; F between =8.46,P ＜ 0.05 ; F intercross group =12.05,P ＜ 0.05).The same trend was seen in night cough symptoms score ((2.12 ± 0.51) vs.(2.38 ±0.38),(1.18±0.44) vs.(1.85±0.49),(1.01 ±0.32) vs.(1.24±0.37) ;Fwithingroup=6.38,P ＜ 0.05 ; F between group =7.59,P ＜ 0.05 ; F intercross group =8.13,P ＜ 0.05).(2) The total efficacy rate of San'ao Pian's was 86.7％,including 22 cases were cured,17 cases markedly effective,higher than that of control group (68.9％),including 14 cases were cured and 17 cases markedly effective,and the difference was significant.Conclusion San'ao Pian's is proved with better clinical effect in terms of treating subacute cough induced by airway inflammation.
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Objective To evaluate the effects of continuous intra-tracheal gas insufflation (TGI) during mechanical ventilation for protecting the juvenile piglets with acute lung injury (ALI) induced by endotoxin. Method Twelve healthy juvenile piglets were anesthetized and mechanically ventilated at 2 cmH2O PEEP with 10 cmH2O peak inspiration pressure. The piglets were challenged with lipopolysaccharide (LPS) and randomly (random number) assigned to two groups (n = 6 each): (1) piglets treated with mechanical ventilation alone (group MV) and (2) piglets treated with TGI by continuous airway flow of 2 L/min (group TGI). FiO2 was set at 0.4 to avoid oxygen toxicity, and the piglets were continuously monitored with an oxygen analyzer. Results Tidal volume, ventilation efficacy index and mean airway pressure were significantly improved in piglets of TGI group (P < 0.01 or P < 0.05). Four hours after ALI, pH decreased to below 7.20 in piglets of MV group, and was higher in piglets of TGI group (P < 0.01). Similarly, PaCO2 was stable and was significantly lower in piglets of TGI group than that in piglets of MV group (P < 0.01). PaO2 and PaO2/FiO2 increased in piglets of TGI group (P < 0.05). There were no significant differences in heart rate, respiraaatory rate, mean arterial pressure, central venous pressure, dynamic lung compliance and mean resistance of airway between two groups. Lung histopathological changes showed severe inflammation,and intra-alveolar hemorrhage and interstitial patchy hemorrhage were ameliorated and the lungs were more homogenously expanded in piglets of TGI group. Conclusions Continuous TGI during MV can significantly improve gas exchange and ventilation efficacy, and may provide a better treatment for acute lung injury.
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BACKGROUND:Traditionally, forced expiratory volume in one second (FEV1) was used to evaluate exercise response of patients with asthma; however, patients obviously had panting after exercise, so FEV1 was affected commonly. Impulse oscillometry (IOS) is a new technique for measuring respiratory impedance that do not require maximal inspiration and forced expiration.OBJECTIVE: To study airway resistance with IOS before and after exercise in healthy and asthmatic patients and investigate the significance of exercise excitation and IOS assessment.DESIGN: Synchronically non-randomized case contrast study.SETTING: Department of Respiratory Medicine, East Hospital Affiliated to Tongji University.PARTICIPANTS: A total of 14 male patients with bronchial asthma who were regarded as the asthmatic group were selected from Department of Respiratory Medicine of Shanghai East Hospital from January to October 2006. They were in a clinical stationary phase. Another 14 male healthy subjects were selected as the control group and ages of all subjects ranged from 29 to 50 years. All subjects provided the confirmed consent.METHODS: IOS was used to measure basic value of respiratory resistance, and then subjects underwent exercise challenge. Nose of subjects was clipped breathing through mouth. Within 3-4 minutes, heart rate was increased to 90% and maintained for 6 minutes during challenge. Respiratory resistance was repeatedly measured at 1, 5, 10, 15 and 20 minutes after exercise, including airway hyperresponse (AHR), total respiratory resistance, central resistance, peripheral resistance and resonance frequency at 5, 20 and 35 Hz of pulse frequency, elasrtic resistance and inertia resistance (X5 and X35) at 5 and 10 Hz of pulse frequency. In addition, difference of AHR at 5 and 35 Hz was calculated, and change ratios of both Rcentral and Rperipheral were calculated as (highest value after exercise-baseline value)/baseline value × 100%.MAIN OUTCOME MEASURES: Basic value of respiratory resistance by using IOS and exercise challenge test.RESULTS: All 14 patients with bronchial asthma and 14 healthy subjects were involved in the final analysis. Peripheral resistance (Rperiphera) was significantly higher than central resistance (Rcentral) in asthmatic patients (P ＜ 0.01). The maximal increase of respiratory impedance occurred from 5 minutes to 10 minutes after exercise in asthmatics. Resonance frequency (Fres) of asthmatics before and after exercise was significantly increased than that of controls (P ＜ 0.01).Change ratios of Fres from asthmatics were higher than that from control group (P ＜ 0.01). After challenge, R5, R5-R20,Zrespir and X5 from asthmatics changed significantly than that from controls (P ＜ 0.01). The increment change value of After exercise Zrespir increased significantly, because obstruction of small bronchi during expiration and impedance increased abruptly. Air trapping was expressed in VT-Zrespir graph in 57.1% patients. There was no difference in the VT-Zrespir graph of controls before and after exercise.CONCLUSION: The main site of airflow obstruction was in small airways in asthmatics after exercise challenge. The general acceptance of IOS method was good among the asthmatic patients. The airway response of exercise challenge may be assessed more accurately with IOS that do not require a maximal inspiration and forced expiration.