1.A study on gyrA mutant in Levofloxacin-resistant Brucella melitensis induced resistant strains in vitro
Chinese Journal of Endemiology 2009;28(5):480-483
Objective To study on the drug-resistance mechanism of Brucella resistance to Quinolone antibiotics to guide the selection and use of antimicrobial agents in clinical practice. Methods Six strains of Brucella melitensis(Bru1, Bru2, Bru3, Bru4, Bru5, Bru6) were selected to be induced resistance to levofloxacin in vitro respectively. The MICs of the 6 strains of Brucella melitensis and induced resistant strains were measured by agar dilution method. The sensitivity to Quinolone antibiotics (Levofloxacin, Ciprofloxacin, Lomefloxacin, Norfloxacin, Fleroxacin, Ofloxaein) of 6 strains of Brucella melitensis and induced resistant strains was measured by K-B method. The gyrA of the 6 strains of Brucella melitensis and induced resistant strains was amplified by PCR, then the nucleotide sequence of the genes were analyzed. Results The MICs of Bru1,Bru2,Bru3,Bru4, Bru6 were 0.50 μg/ml and Bru5 was 0.25 μg/ml. The strains Bin3, Bru4 were induced into drug-resistant strains by Levofloxacin, then were named LEVR3 and LEVR4 respectively. The MICs of LEVR3 and LEVR4 were 64,128 μg/ml with 128 and 256 times higher than that of the parental strains. The 6 strains of Brucella melitensis were sensitive to Quinolone antibiotics, LEVR3 and LEVR4 were resistant to Quinolone antibiotics. Neucleotide sequence analysis and comparison of the derived amino acid sequence revealed that Quinolone resistance-determing region of GyrA had a substitution at position Ala87 and Asp91 in laboratory resistant strains. Conclusion The results of in vitro experiments show that acquired resistance of Brucella melitensis strains to Levofloxacin could beinduced when exposed to high level of some antibacterial agents for short term. Two drug-resistant strains occur mutations in gyrA and have cross-resistance to other Quinolones.
2.Impact of Stimulation Parameters on Event-related Potential P300
Chunyan MAO ; Li SUN ; Yuan YUAN
Chinese Journal of Rehabilitation Theory and Practice 2016;22(7):821-823
Objective To observe the latency and amplitude characteristics of event-related potential P300 under different target stimu-lus. Methods During January to December, 2014, 35 patients with memory impairment received P300 test under the percentage of target stimuli of 0.10 and 0.25. The latency, amplitude and area under amplitude were compared. Results There was no significant difference in la-tency (t<0.256, P>0.800), but the amplitude and area under amplitude increased under 0.10 compared with those under 0.25 (t>4.259, P<0.05). Conclusion Less target stimulation may improve the patients' attention and increase amplitude of P300, but no more interference in la-tency, suggesting latency of P300 is more stable for assessment of cognitive function.
4.Effects of different types of acute respiratory distress syndrome on high frequency oscillatory ventilation-improved extravascular lung water and pulmonary vascular permeability in piglets
Qiujie LI ; Yin YUAN ; Yumei LI ; Leying SUN ; Shiying YUAN
Chinese Journal of Anesthesiology 2013;33(9):1142-1144
Objective To evaluate the effects of different types of acute respiratory distress syndrome (ARDS) on high frequency oscillatory ventilation (HFOV)-improved extravascular lung water and pulmonary vascular permeability in piglets.Methods Twelve healthy piglets,weighing 15-20 kg,were randomly allocated into 2 groups (n =6 each) using a random number table:endogenous ARDS (P group) and exogenous ARDS (EXP group).Anesthesia was induced with midazolam and propofol.The tracheal tube was inserted.Anesthesia was maintained with iv infusion of propofol and fentanyl.ARDS was induced with normal saline infused via the tracheal tube in group P.ARDS was induced with oleic acid 0.05 ml/kg injected intravenously over 45-60 min in group EXP.Then 4 h of HFOV was performed.Before ARDS (T0),immediately after ARDS (Ti) and at 1,2,3 and 4 h of HFOV (T2-5),arterial blood samples were collected for blood gas analysis and the variables such as extravascular lung water index (EVLWI),pulmonary vascular permeability index (PVPI),and extravascular lung water (EVLW)/intra thoracic blood volume (ITBV) were monitored.The oxygenation index (PaO2/FiO2) was calculated.Results Compared with the baseline value at T0,the oxygenation index was decreased at T1,and the oxygenation index was less than 200 mm Hg in the two groups (P < 0.05).There was no significant difference in the maximum degree of changes in EVLWI,PVPI and EVLW/ITBV between the two groups (P > 0.05).Conclusion Endogenous and exogenous ARDS disease factor does not affect HFOV-improved extravascular lung water and pulmonary vascular permeability in piglets.
5.Effect of high-frequency oscillatory ventilation on lung injury in piglets with acute respiratory distress syndrome
Leying SUN ; Yin YUAN ; Shiying YUAN ; Yumei LI ; Qiujie LI
Chinese Journal of Anesthesiology 2016;36(5):629-632
Objective To evaluate the effect of high-frequency oscillatory ventilation (HFOV) on lung injury in the piglets with acute respiratory distress syndrome (ARDS).Methods Twelve male piglets,aged 6-8 weeks,weighing 14-16 kg,were randomly divided into 2 groups (n=6 each) using a random number table:conventional mechanical ventilation with low tidal volume group (CMV group) and HFOV group.ARDS was induced by bilateral pulmonary lavages with isotonic saline (38 ℃),repeated every 10 min until the oxygenation index<200 mmHg.After successful establishment of the model,CMV group was ventilated using conventional mechanical ventilation with low tidal volumes.After successful establishment of the model,HFOV group was ventilated using HFOV,lung recruitment was performed,the airway pressure was set at 25 cmH2O and maintained at this level for 30 s,and the airway pressure was then adjusted 5 cmH2O higher than that after successful establishment of the model,with bias flow 25 L/min,inspiratory time ratio 33%,frequency 8 Hz,amplitude 40-80 cmH2O,and inspiratory oxygen fraction 1.0.In both groups,carbon dioxide partial pressure was maintained between 35 and 50 mmHg.Before establishment of the model (baseline),after successful establishment of the model (T1),and at 0.5,1.0,2.0 and 4.0 h after beginning of mechanical ventilation (T2-5),blood samples were collected from the femoral artery and central vein for blood gas analysis,arterial oxygen partial pressure and carbon dioxide partial pressure were recorded,oxygen delivery index,oxygen consumption index,oxygenation index and intrapulmonary shunt were calculated,and the improvement in pulmonary function (oxygenation index ≥ 200 mmHg) was recorded.At T0,T1 and T5,venous blood samples were collected for determination of the concentrations of serum Clara cell secretory protein 16,soluble intercellular adhesion molecule-1,and high-mobility group box 1.Results Compared with CMV group,the arterial oxygen partial pressure at T35 and oxygenation index at T4.5 were significantly increased (P < 0.05),and no significant change was found in the other parameters in HFOV group (P>0.05).Conclusion Compared with conventional mechanical ventilation with low tidal volumes,although HFOV improves lung oxygenation,the degree is small in the piglets with ARDS.
6.The inlfuence of diagnostic interval and other clinicopathological factors on the prognosis of triple-negative breast cancer
Chu SUN ; Hongtao LI ; Zimei LIU ; Yuan YUAN ; Zan SHEN
China Oncology 2016;26(6):538-545
Background and purpose:The time from ifrst onset of symptoms or signs to a deifnitive diagnosis is deifned as diagnostic interval (DI). The relation of DI to other clinicopathological parameters andthe impact of DI on prognosis of patients with triple-negative breast cancer (TNBC) remain unclear.This article plans to make an intensive study of these questions.Methods:The clinical records of a series of 83 consecutively presenting unselected patients referred to the Shanghai Sixth People’s Hospital with diagnosed TNBC between September 2009 and September 2015 were retrospectively reviewed. Clinical and pathological factors included were investigated by univariate analysis using the Kaplan-Meier method, the factors associated with prognosis were further evaluated by multivariable analysis with Cox progression model.t-test and Kruskal-Wallis test were used to study the correlation between DI and other characters.Results:DI: stage T3>T1 (P=0.01), stageⅢ>Ⅱ (P=0.03) andⅠ (P=0.01). Compared with patients of DI≥3 months, the <3 months group had earlier age (P=0.028) and TNM stage (P=0.035). T stage, N stage, neoadjuvant chemotherapy, TNM stage and DI are inlfuencing factors of overall survival (OS). Age, T stage, N stage, TNM stage, menstrual status and neoadjuvant chemotherapy are inlfuencing factors of progression-free survival (PFS). TNM staging is an independent inlfuencing factor for OS and PFS.Conclusion:Patients with later disease stage were more likely to have a longer DI; The shorter DI, the earlier age and stage of disease; DI is the inlfuence factor of OS; TNM stage is an independent inlfuencing factor for OS and PFS.
7.Protective effect of Ulinastatin on lung pathological changes in paraquat intoxication rats
Mingli SUN ; Haifeng LI ; Yuan YUAN ; Xiaoliang LIU
Chinese Journal of Emergency Medicine 2010;19(12):1291-1295
Objective To evaluate the protective effect of Ulinastatin on pathological changes of lung induced by paraquat (PQ) intoxication in rats. Method Sixty female SD rats were randomly and equally divided into three groups ( n = 20 in each group). On the experimental day, the rats of group A (control group) received 1 mL of saline intragastrically; the rats of group B (PQ, group) received 1 mL of PQ intragastrically (40 mg/kg diluted in saline) and then treated with saline injected intraperitoneally once a day, and the rats of group Group C (Ulinastatin treatment group) received 1 mL of PQ intragastrically (40 mg/kg,diluted in saline) and then treated with 1 mL of Ulinastatin ( 120 000 IU/kg) intraperitoneally twice a day. The rats ( n = 5 in each subgroup) were sacrificed on the 7th, 14th, 21st, and 28th day for observing the histopathological changes of the lung stained with hematoxylin-eosin stain (HE) and Masson's trichrome stain. Data were semi-quantitatively analyzed by SPSS10.0 software. Hierarchical data were analyzed by rank and inspection. The data that have significance analyzed by comparison test (Bonferroni methods). Results Paraquat intoxication resulted in significant alveolitis and lung fibrosis. Ulinastatin treatment alleviated lung edema, hemorrhage, inflammatory cell infiltration, and fiber aggregation. Conclusions These data demonstrated the Ulinastatin exerted protective effects against PQ-induced lung injury. Ulinastatin could to be used in clinical management for PQ intoxication.
8.Intelligence Delay in Children with Cerebral Palsy of Preschool Age
Dianrong SUN ; Mei HOU ; Jun LI ; Yuan LI ; Aijuan SUN
Chinese Journal of Rehabilitation Theory and Practice 2013;19(9):874-877
Objective To investigate the intelligence level and characters in children with cerebral palsy of 4~6 years old, and the factors related with their intelligence. Methods 215 cerebral palsy children of 4~6 years old were assessed with Wechsler Intelligence Scale for children,Gross Motor Function Classification System (GMFCS). Results 210 children finished their intelligence test, and 113 presented mental retardation. The intelligence level was higher in the children with spastic diaplegia, hemiplegia and dyskinetic type than those with quadriplegia and ataxia type. The difference was significant between VIQ and PIQ in the spastic diplegia (P<0.05). The mental retardation was more frequent in the children with GMFCS Ⅳ~Ⅴ, or with epilepsy or microcephaly than those with GMFCS Ⅰ~Ⅲ or without the complications (P<0.05). Conclusion The incidence of mental retardation in children with cerebral palsy was related with the neurotypes, motor function and complications.
10.Effect of berberine on the proliferation and apoptosis of lung cancer stem cells and the possible mechanism
Yanzhen SUN ; Zhen LI ; Zheng YUAN
Chinese Journal of Tissue Engineering Research 2017;21(9):1313-1318
BACKGROUND: Previous studies have demonstrated that berberine represses multiple tumors and tumor stem cells, but the effect of berberine on lung cancer stem cells (LCSCs) remains unclear. OBJECTIVE: To explore the effect of berberine on the proliferation and apoptosis of LCSCs and the possible mechanism. METHODS: CD133+ LCSCs were separated from A549 cells by immunomagnetic beads. The effects of different concentrations (0, 2.5, 5, 10, 20, 40 mg/L) of berberine on the proliferation and apoptosis of LCSCs were determined by MTT and flow cytometry analysis, respectively. In order to further affirm the effect of berberine on the proliferation and apoptosis of LCSCs, the expression levels of Ki67, Bax and Bcl-2 protein were detected by western blot. In addition, to investigate the potential mechanism by which berberine exerts regulatory effects on LCSCs, the expression levels of Hedgehog signaling pathway-associated proteins (PTCH1, SHH, Gli-1 and SMO) were determined. RESULTS AND CONCLUSION: After magnetic cell sorting, the content of the CD133+ fraction was enriched up to 84.13%. MTT and flow cytometry assays showed that berberine inhibited proliferation and promoted apoptosis of LCSCs in a concentration-dependent manner. Western blot analysis showed that the expression levels of Ki67, Bcl-2, PTCH1, SHH, Gli-1 and SMO proteins of LCSCs cultured in the medium with 20 mg/L berberine were dramatically decreased compared to the control, while the expression level of Bax protein was markedly increased compared to the control. These findings suggest that berberine may inhibit proliferation and promote apoptosis for LCSCs through the Hedgehog signaling pathway.BACKGROUND: Previous studies have demonstrated that berberine represses multiple tumors and tumor stem cells, but the effect of berberine on lung cancer stem cells (LCSCs) remains unclear. OBJECTIVE: To explore the effect of berberine on the proliferation and apoptosis of LCSCs and the possible mechanism. METHODS: CD133+ LCSCs were separated from A549 cells by immunomagnetic beads. The effects of different concentrations (0, 2.5, 5, 10, 20, 40 mg/L) of berberine on the proliferation and apoptosis of LCSCs were determined by MTT and flow cytometry analysis, respectively. In order to further affirm the effect of berberine on the proliferation and apoptosis of LCSCs, the expression levels of Ki67, Bax and Bcl-2 protein were detected by western blot. In addition, to investigate the potential mechanism by which berberine exerts regulatory effects on LCSCs, the expression levels of Hedgehog signaling pathway-associated proteins (PTCH1, SHH, Gli-1 and SMO) were determined. RESULTS AND CONCLUSION: After magnetic cell sorting, the content of the CD133+ fraction was enriched up to 84.13%. MTT and flow cytometry assays showed that berberine inhibited proliferation and promoted apoptosis of LCSCs in a concentration-dependent manner. Western blot analysis showed that the expression levels of Ki67, Bcl-2, PTCH1, SHH, Gli-1 and SMO proteins of LCSCs cultured in the medium with 20 mg/L berberine were dramatically decreased compared to the control, while the expression level of Bax protein was markedly increased compared to the control. These findings suggest that berberine may inhibit proliferation and promote apoptosis for LCSCs through the Hedgehog signaling pathway.