Objective To explore the adeno-associated virus (AAV)gene transduction and cellular endocytosis mediated by ultrasound combined with microbubbles in two types of cells.Methods HeLa and NIH/3T3 cells were infected by rAAV2-EGFP at a concentration gradient to get the optimal concentrations for enhancement.At these concentrations,HeLa and NIH/3T3 cells were infected by rAAV2-EGFP mediated by ultrasound combined with microbubbles.The gene transduction efficiency were observed and measured by fluorescence microscopy and flow cytometry at 48 h after treatment.The cell viability was tested by CCK-8.The number and distribution of cellular clathrin-coated endocytic pits were observed by confocal fluorescence microscopy and transmission electron microscopy on 45 min after treatment.Results The optimal concentrations for HeLa and NIH/3T3 cells were 2000 v.g./cell and 10000 v.g./cell.Ultrasound combined with microbubbles significantly enhanced the transduction efficiency of rAAV2-EGFP (P <0.01) without significant cell viability decrease (P > 0.05 ).Confocal fluorescence microscopy and transmission electron microscopy demonstrated that clathrin-coated endocytic pits were more obviously increased in ultrasound combined with microbubbles mediated AAV transduction group than AAV transduction group. Conclusions Ultrasound combined with microbubbles can efficiently enhance the gene transduction of AAV,whose cellular transportation depends on cellular endocytosis,in two types of cells.Stimulating cellular endocytosis might be one of the mechanisms of enhanced cellular transportation of AAV mediated by ultrasound combined with microbubbles.

Objective To investigate the factors affecting the susceptibility of tigecycline and assess the testing methods.Methods The 116 isolates of Acinetobacter baumanaii and Klebsiella pneumoniae were collected in 13 hospitals from January to December,2010,to evaluate the effects on the tigecycline susceptibility of the overnight medium,medium brand and lot number,respectively.The 56 isolates of Acinetobacter baumannii and the 47 isolates of Klebsiella pneumoniae were selected randomly according to the MIC distribution proportion in 2010 and 2012.The broth microdilution was taken as the reference method to evaluate the effects of the agar dilution,disk diffusion,MIC Test Strip (MTS) and Vitek2 (GN16) on the susceptibility of tigecycline.Results The essential agreement (EA) of Acinetobacter baumannii and Klebsiella pneumoniae is 89.7％ (52/58)and 87.9％ (51/58) using overnight medium and fresh medium respectively.Both EA and categorical agreement (CA) of the different brands (BBL and Oxoid) and lot numbers are 100％ using agar dilution.According to the FDA break point criteria,the CA/EA is 77.7％ (80/103)/99.0％ (102/103),87.4％ (90/103)/98.1％ (101/103),64.1％ (66/103)/76.7％ (79/103) using agar dilution,MTS,Vitek (GN16) with respect to broth microdilution.The CA is 79.6％ (82/ 103,S≥14 mm,R≤10 mm),69.9％ (72/103,S≥ 16 mm,R≤ 12 mm),34.0％ (35/103,S≥19 mm,R≤14 mm)using disk diffusion method compared with broth microdilution (FDA break point criteria).Conclusions The susceptibility of tigecycline must be tested using fresh medium.The medium brands and lot numbers used in this test have no effects on the tigecycline susceptibility to Acinetobacter baumannii and Klebsiella pneumoniae.There exist the better correlations on MIC using agar dilution and MTS than the disk diffusion and Vitek(GN16) compared with broth microdilution.It is expected that the consistency can be improved by adjusting the break point of disk diffusion.

Objective To evaluate the clinical performance of the BACTEC Plus aerobic,BACTEC Lytic anaerobic,BacT/Alert aerobic and anaerobic blood culture media in detection of bloodstream infections.Methods Retrospective study was conducted.A total of four blood culture bottles from each inpatient with suspected bloodstream infections were collected and analyzed from June 2013 to September 2015 in Peking University People's Hospital.The four bottles,including BACTEC Plus aerobic,BACTEC Lytic anaerobic,BacT/Alert FA aerobic and BacT/Alert FN anaerobic media,and was incubated for 5 days in the BacT/Alert 3D and BACTEC FX instruments,respectively.Time to detection (TTD) and positive rate in detecting bacteria of the two systems were evaluated by Wilcoxon test and Chi-square test.Results Among 2 189 total cultures collected,20 were excluded because of blood shortage and 201 (9.27％) were positive for pathogens.The positive rates of BACTEC Plus aerobic media and BacT/Alert FA aerobic media were 75.3％ (140/186) and 69.4％ (129/186) (x2 =1.625,P=0.202),respectively.While,the positive rates of BacT/Alert FN anaerobic media and BACTEC Lytic anaerobic media were 81.8％ (99/121) and 63.6％ (77/121) for total organisms,respectively (x2 =10.083,P =0.001).A significant difference in TTD was detected in BACTEC Plus aerobic media[11.0 (8.0-16.0) h] and BacT/Alert FA aerobic media[13.9 (10.4-18.7) h] (Z =-5.240,P ＜ 0.001).BACTEC Lyric anaerobic media[8.0(7.0-10.0) h] had a shorter TTD (Z =-4.299,P ＜ 0.001) than BacT/Alert FN anaerobic media[11.3(9.3-12.7) h].The positive rates of BACTEC and BacT/Alert system were 74.13％ (149/201) and 74.63％ (150/201),respectively,compared with taking one set from each system.Conclusions BACTEC media has a shorter TTD and almost the same bacterial recovery,and lower false positive rate than the BacT/ Alert media.

).The complication rate in RIVVH was lower than that in the control group (P<0.05).Conclusions Early RIVVH was effective in the treatment of SAP,and may be an option as adjuvant treatment measure.

Objective To observe the effect of deep muscle relaxation by rocuronium on oxygenation of normal frequency jet ventilation during rigid bronchoscopy procedures.Methods From December 2021 to February 2023,60 patients with central airway diseases underwent rigid bronchoscopy under general anesthesia,they were randomly divided into deep muscle relaxation group(group D,n = 30)and moderate muscle relaxation group(group M,n = 30).After induction of general anesthesia,the patients were inserted rigid bronchoscopy for jet ventilation,muscle relaxant was rocuronium in induction and maintenance.Train of four(TOF)stimulation was used to measure the depth of muscle relaxation in group M,and the TOF count was maintained at 1 or 2;In Group D,the depth of muscle relaxation was measured by post tetanic count(PTC),and the PTC was maintained at 1 or 2.After operation,Sugammadex antagonized residual muscle relaxation.Results There was no significant difference in operation time,recovery time and extubation time between the two groups(P＞0.05).The total operation time,operation pause time and anesthesia time in group D were shorter than those in group M,the dosage of muscle relaxant in group D was more than that in group M,the incidence of hypoxemia during surgery in group D was less than that in group M,the operators'satisfaction in group D was better than that in group M,and the arterial partial pressure of oxygen(PaO2)in group D was higher than that in group M at 15 min(T1)and 30 min(T2)after jet ventilation,the number of patients with postoperative sore throat in group D was less than that in group M,the differences were statistically significant(P＜0.05).Conclusion The application of deep muscle relaxation by rocuronium in rigid bronchoscopy procedures can improve the oxygenation effect of normal frequency jet ventilation,reduce the operation pause time and anesthesia time,improve the satisfaction of operators,antagonizing residual muscle relaxation with Sugammadex can relieve the worry of delayed recovery from deep muscle relaxation.

To present the clinical characteristics and the misdiagnosis rate of acute coronary syndrome manifested primarily as throat discomfort, we conducted a multicentric and retrospective study in the cardiology and otorhinolaryngology departments. Records of patients with primary complaint of throat discomfort, absence of chest pain at onset, and an ultimate diagnosis of acute coronary syndrome, as well as patients with pharyngitis (as controls) were collected from May 2015 to April 2016. The patients' main manifestations were compared. Logistic regression results showed that chest tightness, dyspnea, perspiring, and exertional throat symptoms were significantly associated with acute coronary syndrome, with odds ratios of 8.3 (95% CI 2.2-31.5), 10.9 (95% CI 1.8-66.9), 25.4 (95% CI 3.6-179.9), and 81.2 (95% CI 13.0-506.7). A total of 25 (56.82%) out of 44 acute coronary syndrome patients, who were first admitted to the otorhinolaryngology department, were misdiagnosed, with a 12% (3/25) mortality rate. Throat discomfort can be the principal manifestation of acute coronary syndrome. Such patients exhibit high misdiagnosis and mortality rates. Exertional throat symptoms, chest tightness, perspiring, and dyspnea were important indicators of acute coronary syndrome in patients whose main complaint was throat discomfort. The awareness of this condition will result in prompt diagnosis and reduce morbidity and mortality.
Acute Coronary Syndrome/etiology* ; Dyspnea/etiology* ; Humans ; Pharyngitis/diagnosis* ; Pharynx ; Retrospective Studies

With the accelerated aging society in China, the incidence of biliary surgical diseases in the elderly has increased significantly. The clinical characteristics of these patients indicate that improving treatment outcomes and realizing healthy aging are worthy of attention. How to effectively improve the treatment effect of geriatric biliary surgical diseases has attracted widespread attention. This paper reviews and comments on the hotspots and difficulties of biliary surgery in older patients from six aspects: (1) higher morbidity associated with an aging society, (2) prevention and control of pre-operative risks, (3) extending the indications of laparoscopic surgery, (4) urgent standardization of minimally invasive surgery, (5) precise technological progress in hepatobiliary surgery, and (6) guarantee of peri-operative safety. It is of great significance to fully understand the focus of controversy, actively make use of its favorable factors, and effectively avoid its unfavorable factors, for further improving the therapeutic effects of geriatric biliary surgical diseases, and thus benefits the vast older patients with biliary surgical diseases. Accordingly, a historical record with the highest age of 93 years for laparoscopic transcystic common bile duct exploration has been created by us recently.
Humans ; Aged ; Aged, 80 and over ; Biliary Tract Surgical Procedures ; Gallstones ; Laparoscopy ; Treatment Outcome ; Aging ; Retrospective Studies