Objective To investigate the effects of doublecortin-like kinase 1(DCLK1)on the malignant biological behaviors,such as proliferation,migration,and invasion,of A549 cell line and their corresponding mechanisms.Methods DCLK1-overexpressing A549 cell lines were established through lentiviral infection,and DCLK1 expression was validated by using RT-PCR and Western blot analysis.Proliferation ability was assessed with CCK-8 and plate cloning assays,and migration and invasion abilities were examined with Transwell assays.The pathway regulated by DCLK1 in lung adenocarcinoma was analyzed on the basis of the TCGA lung adenocarcinoma cohort with pathway enrichment analysis and verified through Western blot analysis.Results DCLK1 overexpression in A549 cells promoted cell proliferation,migration,and invasion.The inhibition of the FAK/PI3K/AKT/mTOR signaling pathway impaired the DCLK1-mediated malignant behavior of A549 cells.Conclusion DCLK1 promotes the malignant behavior of A549 cells through the activation of the FAK/PI3K/AKT/mTOR signaling pathway.

Objective:To explore the clinical utility of metagenomic next-generation sequencing (mNGS) for patients with critically ill atypical rickettsial infections in the early diagnosis and therapy.Methods:From Jan 2020 to Aug 2022, clinical features, blood biochemical results, imaging data and mNGS results in patients with unexplained critical illnesses were collected and analyzed retrospectively. Fisher's exact test was used to compare the positive rate of mNGS and weil felix reaction.Results:All 15 patients with rickettsial disease had fever, 12 cases had headache, but only 3 had a typical rash or scab of diagnostic significance, 6 had septic shock and all had multi-organ dysfunction; blood mNGS tests were positive in 15 cases, of which 10 had Orientia tsutsugamushi detected in their blood and the remaining five had Rickettsia moschata detected in their blood. The positive rate of mNGS was significantly higher than that of the weil felix reaction (15/15 vs 0, P<0.001). All patients were given doxycycline and other treatments after diagnosis, of which 14 improved and were discharged, and one died 1 week after discharge due to critical condition and abandonment of treatment. Conclusion:mNGS can improve the detection rate of atypical rickettsiae in patients with negative routine test results, which can provide valuable reference basis for early diagnosis and early anti-infection treatment of patients with critical rickettsial disease.

Objective:To investigate the safety and efficacy of stent-assisted coil embolization in acute intracranial ruptured wide-necked aneurysms, and explore the safety of different antiplatelet drug regimens during perioperative period and the risk factors for prognoses.Methods:A perspective multicenter study was performed. A total of 417 patients with intracranial ruptured wide-necked aneurysms treated by stent-assisted coil embolization in Neurosurgery Departments of 4 hospitals (First Affiliated Hospital of Wenzhou Medical University [ n=41], Zhujiang Hospital of Southern Medical University [ n=111], Tangdu Hospital of Air Force Military Medical University [ n=100], and Xuanwu Hospital of Capital Medical University [ n=165]) from June 2017 to January 2020 were included. According to the different antiplatelet drugs regimens used in perioperative period, these patients were divided into loaded clopidogrel group ( n=87), loaded clopidogrel combined with aspirin group ( n=212), and tirofiban group ( n=118). Clinical data and perioperative complications of 3 groups were compared. Modified Rankin scale (mRS) was used to evaluate the prognoses of patients at discharge; differences of clinical data between the poor prognosis group and good prognosis group were compared. Independent risk factors for prognoses of patients with intracranial ruptured wide-necked aneurysms were analyzed by multivariate Logistic regression analysis. Receiver operating characteristic (ROC) curve was used to analyze the predictive values of risk factors in poor prognosis. Results:Degrees of aneurysm embolization immediately after surgery: Raymond grading I was noted in 351 patients (84.2%), grading II in 44 patients (10.6%), and grading III in 22 patients (5.2%). Perioperative complications were noted in 44 patients (10.6%), and death was noted in 4 (1%). Intraoperative thrombosis incidence of the loaded clopidogrel group, loaded clopidogrel combined with aspirin group and tirofiban group was 5.7% (5/87), 5.7% (12/212) and 0.8% (1/118); that in tirofiban group was significantly lower than that in the loaded clopidogrel group and loaded clopidogrel combined with aspirin group ( P<0.05). At discharge, 360 patients (86.3%) had good prognosis and 57 patients (13.7%) had poor prognosis. Multivariate Logistic regression analysis showed age≥60 years ( OR=3.407, 95% CI: 1.620-7.166, P=0.001), preoperative Hunt-Hess grading 3 ( OR=11.445, 95% CI: 3.584-36.547, P<0.001), preoperative Hunt-Hess grading 4 ( OR=88.951, 95% CI: 14.519-544.948, P<0.001), preoperative Hunt-Hess grading 5 ( OR=64.949, 95% CI: 12.809-329.325, P<0.001), and multiple stenting ( OR=4.709, 95% CI: 1.215-18.248, P=0.025) were independent risk factors for poor prognosis of these patients. ROC curves showed that area under the curve of combination of age, number of implanted stents, and preoperative Hunt-Hess grading in predicting poor prognosis of these patients was 0.821, with optimal diagnostic threshold of 0.500, sensitivity of 0.667, and specificity of 0.833. Conclusion:Stent-assisted coil embolization is safe and effective in acute intracranial ruptured wide-necked aneurysms; tirofiban is safe as perioperative antiplatelet drug; patients with old age, preoperative Hunt-Hess grading≥3, and multiple stents are prone to have poor prognosis.

Objective:To investigate the relationship between double-triggering and abnormal movement of air in the lungs (pendelluft phenomenon) under pressure support ventilation (PSV).Methods:A prospective observational study was conducted, postoperative patients admitted to department of critical care medicine of Beijing Tiantan Hospital, Capital Medical University from April 1, 2019 to August 31, 2020 and received invasive mechanical ventilation with PSV mode were enrolled. Electrical impedance tomography (EIT) monitoring was performed. Airway pressure-time, flow-time, global and regional impedance-time curves were synchronously collected and analyzed offline. The volume of abnormal movement of air in the lungs at the beginning of inspiration was measured and defined as pendelluft volume. Double-triggered breaths were identified by trained researchers. Pendelluft volume during double-triggering was measured including the first triggered breath, the double-triggered breath, and the breath immediately following the double-triggered breath. Pendelluft volume was also measured for normal breath during the study. According to the frequency of double-triggering, patients were divided into severe (≥1 time/min) and non-severe double-triggering group. Pendelluft volume, parameters of respiratory mechanics, and clinical outcomes between the two groups were compared.Results:In 40 enrolled patients, a total of 9 711 breaths [(243±63) breaths/patient] were collected and analyzed, among which 222 breaths (2.3%) were identified as double-triggering. The Kappa of interobserver reliability to detect double-triggering was 0.964 [95% confidence interval (95% CI) was 0.946-0.982]. In 222 double-triggered breaths, pendelluft volume could not be measured in 7 breaths (3.2%), but the pendelluft phenomenon did exist as shown by opposite regional impedance change at the beginning of double-triggered inspiration. Finally, pendelluft volume was measured in 215 double-triggered breaths. Meanwhile, 400 normal breaths (10 normal breaths randomly selected for each patient) were identified as control. Compared with normal breath, pendelluft volume significantly increased in the first breath, the double-triggered breath, and the following normal breath [mL: 3.0 (1.4, 6.4), 8.3 (3.6, 13.2), 4.3 (1.9, 9.1) vs. 1.4 (0.7, 2.8), all P < 0.05]. Patients in severe double-triggering, pendelluft volume of normal breath and double-triggered breath were significantly higher than those in non-severe double-triggering group [mL: 1.8 (0.9, 3.2) vs. 1.1 (0.5, 2.1), P < 0.001; 8.5 (3.9, 13.4) vs. 2.0 (0.6, 9.1), P = 0.008]. Patients in severe double-triggering group had significantly higher respiratory rate than that in the non-severe double-triggering group (breaths/min: 20.9±3.5 vs. 15.2±3.7, P < 0.001). There were no significant differences in other respiratory mechanics parameters and main clinical outcomes between the two groups. Conclusions:During PSV, the abnormal movement of air in the lungs (pendelluft phenomenon) was more likely to occur in double-triggering especially in double-triggered breath. The more frequent the double-triggering occurred, the more serious the pendelluft phenomenon was. A higher pendelluft volume of normal breath and a higher respiratory rate were related to severity of double-triggering.

Objective:To develop scientific and practical sensitive indicators with its own characteristics for nursing quality of breast specialty.Methods:A research team was established in August 2018, which searched the databases both at home and abroad, screened and identified relevant literatures according to John Hopkins' evidence level and quality evaluation standards. A sensitive indicator framework for nursing quality of breast specialty was preliminarily formed based on the indicators involved in the literatures. In January 2019, the expert correspondence questionnaire was prepared. Twenty experts were randomly selected from the Qingdao Medical Doctors Association and the Nursing Experts Committee and received two rounds of correspondence consultations using the Delphi method in order to build, modify and confirm the sensitive indicator system for nursing quality of breast specialty.Results:The authority coefficients of the experts in the two rounds of consultation were 0.815 and 0.858; the expert authority coefficient was> 0.80; and the coefficient of variation of each indicator was< 0.30. Eleven sensitive indicators for nursing quality of breast specialty including the awareness rate of functional exercise of the affected limb and lymphedema prevention measures were developed.Conclusions:The sensitive indicators of nursing quality of breast specialty based on the Delphi method is highly practical and credible, which have distinctive specialty characteristics, may be used for the overall care quality assessment and management of breast disease patients, and provide accurate and scientific evaluation for clinical nursing work.

This article interpretsthe expert consensus on venous catheter maintenance, including flushing and locking tube, dressing replacement and catheter maintenance, infusion connectors, catheter removal, education and training, infection control, etc. It is hoped that the expert consensus can provide a reference for clinical nurses to standardize the operations of intravenous catheter maintenance, thereby benefiting patients.

Objective: To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features. Methods: The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups. Results: The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05). Conclusion The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.

Objective To observe and compare the clinical efficacies between electroacupuncture and warm needling in treating low back pain.Method Seventy-eight eligible low back pain patients were randomized into group A of 28 cases, group B of 26 cases, and group C of 24 cases. Group A was intervened by electroacupuncture, group B was by warm needling, and group C was by medication. The short-form McGill Pain Questionnaire, Japanese Orthopaedic Association Scores (JOA), and Oswestry Disability Index were observed before and after treatment, and the therapeutic efficacies were compared.Result In group A, the McGill item scores [Sensory Pain Rating Index (S-PRI), Affective Pain Rating Index (A-PRI)] respectively after 1-week and 2-week treatment as well as in the 1-month and 3-month follow-up were significantly different from that before treatment (P<0.01,P<0.05). In group B and C, the McGill item scores after 2-week treatment and in the 1-month and 3-month follow-up were significantly different from that before treatment in the same group (P<0.01,P<0.05). The JOA and Oswestry scores were significantly changed respectively after 1-week and 2-week treatment and in the 1-month and 3-month follow-up in the three groups compared with that before treatment (P<0.05,P<0.01). After 1-week and 2-week treatment and in the 1-month and 3-month follow-up, the JOA and Oswestry scores in group A were significantly different from that in group C (P<0.05,P<0.01). In the 1-month and 3-month follow-up, the JOA scores in group B were significantly different from that in group C (P<0.05). The total effective rate was 85.7% in group A and 73.1% in group B, both significantly higher than 58.3% in group C (P<0.05). Conclusion Electroacupuncture and warm needling both can produce a significant efficacy in treating low back pain, but warm needling acts comparatively slowly and is less safe.

OBJECTIVE:To establish the method for the determination of tigecycline (TGC) in human plasma. METHODS:After precipitated by acetonitrile,the plasma sample was determined by LC-MS/MS. Using d9-TGC as internal standard,Kromasil C18 column was used with mobile phase consisted of water (containing 0.05% TFA)-acetonitrile (gradient elution) at flow rate of 0.6 ml/min,column temperature of 40 ℃. The ion transitions were performed under ESI positive MRM model at m/z 586.3→513.2 and m/z 595.3→514.3 for TGC and internal standard,respectively. RESULTS:The linear range of TGC was 25-2 000 ng/ml (r=0.999 8),and lowest quantification limit was 25 ng/ml;intra-day and inter-day RSD was 3.15%-7.23%,and relative error was-4.53%-10.48%. Plasma sample kept stable after 3 times of freezing and thawing cycle,at room temperature for 24 h,in automat-ic sample injector for 24 h and freezing for 42 d (RSD<15%). Plasma concentration of TGC was 0-438.0 ng/ml in one patient with pan-drug resistant bacteria infection(0-12 h after administration). CONCLUSIONS:The developed method is accurate,sensi-tive and specific,and can be used for plasma concentration determination of TGC and pharmacokinetic study.

Objective To measure the organ weights , physiological and biochemical parameters and immune cells of NOD/SCID mice.Methods Mice at five and ten weeks of age were selected .The organ weights, blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in NOD/SCID mice at six week of age in terms of its T , B lymphocyte function and NK cell activity.Results Among the same sex group of NOD/SCID mice, the weights of kidney, liver, heart and lung, and the blood physiological indexes of RBC , HGB, HCT, MCV, MCH, RDW, and the blood biochemical indexes of TP , ALB, ALP, CHO, TBIL show significant difference between 5 and 10 weeks.At the same age, HCT, GLOB, A/G, CHO, TG, TBIL and the UN are significantly different between male and female NOD /SCID mice.NOD/SCID mice lack T cells (0.37 ±0.26)%、CD4+T cells (0.35 ±0.13)%、CD8 +T cells (0.47 ±0.10)%、CD19+B cells (0.13 ± 0.05)%、B220+B cells (1.20 ±0.44)%.The percentage of NK cells is (6.90 ±0.82)%, and the percentage of granulocytes is (47.88 ±15.54)%.Conclusions The study indicates that NOD/SCID mice show the deficiency of T , B and NK cells function .The organ weights , blood physiological and biochemical parameters are affected by age and gender . The studied parameters of NOD/SCID mice are similar with the same strain in other countries .