Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .

Objective To investigate the utility value of monoexponential and biexponential DWI in the differential diagnosis between benign and malignant liver neoplasms.Methods Seventy three patients with pathologically or clinically confirmed liver mass,were analyzed retrospectively and categorized into benign and malignant groups between January 2013 and October 2013.Malignant group included 46 patients with 53 lesions,while 27 patients in benign group had 35 lesions.All patients underwent MR examinations on 3.0T system (GE 750).Conventional MR T1WI,T2WI,DWI(b=0,800 s/mm2) (to obtain ADC with monoexponential modeling),multi-b value DWI(b=0,20 50,100,200,400,600,800 and 1 200 s/mm2) (to obtain Slow-ADC,Fast-ADC,f with biexponential modeling) and dynamic enhancement were performed.The ADC,Slow-ADC,Fast-ADC and f mean values of benign and malignant liver neoplasms were measured and analyzed by using independent samples t test.Diagnostic efficacy of these parameters in malignant group was evaluated by using receiver operating characteristic curve,with histopathologic findings as the gold standard.Results ADC,Slow-ADC,Fast-ADC and f of malignant group were lower than those of benign group [ADC:(1.79±0.35)× 10-3 mm2/s vs (1.16±0.36) × 10-3 mm2/s; Slow-ADC:(1.67±0.25) × 10-3 mm2/s vs(0.94±0.22)×10-3mm2/s; Fast-ADC(72.40±23.70)×10-3mm2/s vs(34.62±17.43)×10-3mm2/s; and f:(33.59± 11.77)％ vs (22.28±8.97)％ in benign and malignant groups,respectively).Significant inter-group difference was observed in ADC,Fast-ADC,Slow-ADC and f (t=0.89,14.77,8.96 and 5.47,respectively and P＜0.05).The areas under the ROC curve (AUC) of ADC,Slow-ADC,Fast-ADC and fwere 0.938,0.974,0.895 and 0.789,respectively.The sensitivity and specificity of ADC,Slow-ADC,Fast-ADC and fwere 90.6％ (48/53),96.2％ (51/53),90.6％ (48/53) and 90.6％ (48/53) and 85.7％ (30/35),91.4％ (32/35),82.9％ (29/35) and 57.1％ (20/35)respectively for differentiating benign from malignant hepatic lesions.Conclusion ADC obtained with mono-exponential modeling and Fast-ADC,Slow-ADC,f obtained with biexponential modeling are useful parameters in distinguishing benign and malignant hepatic lesions,among which slow-ADC demonstrates the highest diagnostic efficacy.

Objective To investigate the effect of hyperoside on proliferation and killing activity of NK cells against pancreatic cancer PANC1 cells in vitro,and explore its potential mechanism.Methods Peripheral blood mononuclear cells of healthy donors were isolated,NK cells were induced with medium contained with IL-2 and different concentrations of hyperoside (0.3,1.6,8,40 and 200 μg/ml) for 12 days.Cell viability was observed by trypan blue staining.Phenotype and perforin,granzyme B expression of NK cells were detected by flow cytometry.Killing activity of NK cells against PANC1 cells were analyzed with lactate dehydrogenase (LDH) releasing method.Results The proportion of NK cells in control group and experimental group treated with different concentration of hyperoside both reached about 80％,respectively.The proliferation of CDs-CD56 + NK cells treated by hyperoside at 0.3,1.6 and 8 μg/ml was (93.76 ±8.77),(106.67 ± 12.35) and (118.50 ± 11.51) times,respectively,which were significantly higher than (73.70 ± 9.43) times of the control group.The expressions of perforin in NK cells treated with hyperoside at 1.6,8 and 40 μg/ml were significantly higher than those of the control group [(82.34 ± 2.90) ％,(89.15 ±3.54) ％,(81.78 ± 2.81)％ vs (72.93 ± 2.06)％].The expressions of granzyme B in NK cells treated with hyperoside at 1.6 and 8 μg/ml were significantly higher than those of the control group [(87.30 ± 1.70) ％,(92.16 ±3.05)％ vs (82.35 ±2.73)％].The killing activity of NK cells against PANC1 cells treated by hyperoside at 1.6 and 8 μg/ml was significantly higher than those of the control group [(63.18 ± 3.77)％,(65.34 ± 4.97) ％ vs (52.16 ± 5.48) ％].The differences were statistically significant (all P ＜ 0.05).Conclusions Hyperoside could promote the proliferation of NK cells at certain concentrations and maybe enhance the killing effect against pancreatic cancer PANC1 cells through up-regulating the expression of perforin and granzyme B in NK cells.

Acute respiratory and circulatory disorders are the most common critical syndromes, the essence of which is damage to the organs/systems of the heart and lungs. These comprise the essential manifestation of disease and injury progression to the severe stage. Its development involves the following components: individual specificity, primary disease strike, dysregulation of the host′s response, and systemic disorders. Admission for acute respiratory and circulatory disorders is a clinical challenge. Based on a previously proposed flow, a critical care ultrasound-based stepwise approach (PIEPEAR) as a standard procedure to manage patients with acute cardiorespiratory compromise and practical experience in recent years, a modified seven-step analysis and treatment process has been developed to help guide clinicians with rational thinking and standardized treatment when faced with acute respiratory and circulatory disorders. The process consists of seven steps: problem-based clinical analysis, intentional information acquisition, evaluation of core disorder based on critical care ultrasound, pathophysiology and host response phenotype identification, etiology diagnosis, act treatment through pathophysiology-host response and etiology, and re-check. The modified seven-step approach is guided by a “modular analysis” style of thinking and visual monitoring. This approach can strengthen the identification of clinical problems and facilitate a three-in-one analysis. It focuses on pathophysiological disorders, body reactions, and primary causes to more accurately understand the condition′s key points, and make treatment more straight forward, to finally achieve the aim of “comprehensive cognition and refined treatment”.

Objective:To investigate the effects of microRNA(miR)-133b on epithelial-mesenchymal transition(EM)of human small airway epithelial cells induced by cigarette smoke extracts(CSE)and its regulatory mechanisms.Methods:The miR-expression profiles with microarray in airway epithelial cells of patients with chronic obstructive pulmonary disease were searched in the Gene Expression Omnibus(GEO)database, and the differentially expressed miRs were searched and verified by a real-time fluorescence quantitative method(qRT-PCR). Human small airway epithelial cells(HSAEpiC)were divided into the control group, the CSE group, the CSE+ miR-133b inhibitor transfection group(inhibitor group)and the CSE+ miR-133b inhibitor negative control transfection group(inhibitor control group)according to different intervention methods.Levels of miR-133b and mRNA levels of transforming growth factor(TGF)-β1, Smad2, E-cadherin and vimentin were detected by RT-PCR; Protein levels of E-cadherin and vimentin were detected by enzyme-linked immunosorbent assays(ELISA)and Western blotting.Results:Nine differentially expressed miRs were found in GSE53519, with miR-133b showing the most significant differential in thee HSAEpiC cell model after verification.CSE induced morphological changes in HSAEpiC cells, and miR-133b inhibitors could partially reverse the morphological changes in cell mode.mRNA and protein expressions of E-cadherin were decreased and expression of Vimentin mRNA and protein were increared in CSE induced HSAEpiC cells( F=9.09、12.35、7.57、101.87, P=0.015、0.007、0.023、0.000); miR-133b inhibitors partally reversed the mRNA and protein expressions of E-cadherin and Vimentin( F=40.59、27.74、15.87、20.42, P=0.000、0.001、0.004、0.002). CSE induced incresed expression of TGF-β1 mRNA and Smad mRNA in HSAEpiC cells, and miR-133b inhibitors partially reversed the changes in TGF-β1 mRNA and Smad mRNA( F=17.25、64.15, P=0.003、0.000). Conclusions:miR-133b may regulate CSE-related HSAEpiC cell EMT through the TGF-β1/Smad pathway.

Purpose To investigate the clinicopathological features,immunophenotype,molecular changes,differential di-agnosis,treatment and prognosis of the micropapillary subtype of serous borderline tumor(MSBT)in the ovary.Methods The clinical and pathological data of 14 cases of ovarian MSBT.Im-munohistochemical EnVision staining was used to detect the ex-pression of IMP3.BRAF and KRAS mutations were detected by qRT-PCR and Sanger sequencing,respectively.Its clinical and pathological characteristics were analyzed with review of relevant literature.Results The age of the patients ranged from 27 to 56 years,with mean 41.7 years.Nine cases had bilateral ovari-an masses.Preoperative serum CA125 increased in 11 cases.On gross examination,the cut section was cystic and solid with intracystal papillae.Microscopically,all cases showed a papilla-ry structure,with the characteristic elongated micropapillae radi-ating directly from the cyst wall or large unbranched papillae.The length to width ratio of the papillae was greater than 5.The cells covering the papillae were cubic to polygonal.Mild to mod-erate atypia was noted with a range of＞5 mm in the micropapil-lary area.Five cases had microinvasion.Six cases had non-in-vasive peritoneal seeding.Five cases were accompanied by asci-tes,and atypical tumor cells were observed in ascites.Three ca-ses had lymph node involvement.Nine cases had psammoma bodies.Immunohistochemically,the tumor cells were positive for ER,PR,CA125,CK7 and WT-1;p53 was wild type,HER2 and IMP3 were negative,and Ki67 was positive in 5％to 30％.KRAS mutations were detected in 3 of 14 cases,inclu-ding G12C,G12D and Q70(nonsense mutation).No BRAF V600E mutation was detected,and 1 case had BRAF T559I mu-tation.Seven patients underwent radical surgery and 7 patients underwent conservative surgery without special treatment after surgery.Five patients had a history of recurrence,and the fol-low-up time ranged from 1 to 12 years.Conclusion MSBT has special morphology,often bilateral,and is prone to peritoneal implantation and recurrence.It should be distinguished from classical ovarian serous borderline tumor.

Critical ultrasonography(CUS) is different from the traditional diagnostic ultrasound,the examiner and interpreter of the image are critical care medicine physicians.The core content of CUS is to evaluate the pathophysiological changes of organs and systems and etiology changes.With the idea of critical care medicine as the soul,it can integrate the above information and clinical information,bedside real-time diagnosis and titration treatment,and evaluate the therapeutic effect so as to improve the outcome.CUS is a traditional technique which is applied as a new application method.The consensus of experts on critical ultrasonography in China released in 2016 put forward consensus suggestions on the concept,implementation and application of CUS.It should be further emphasized that the accurate and objective assessment and implementation of CUS requires the standardization of ultrasound image acquisition and the need to establish a CUS procedure.At the same time,the standardized training for CUS accepted by critical care medicine physicians requires the application of technical specifications,and the establishment of technical specifications is the basis for the quality control and continuous improvement of CUS.Chinese Critical Ultrasound Study Group and Critical Hemodynamic Therapy Collabration Group,based on the rich experience of clinical practice in critical care and research,combined with the essence of CUS,to learn the traditional ultrasonic essence,established the clinical application technical specifications of CUS,including in five parts:basic view and relevant indicators to obtain in CUS;basic norms for viscera organ assessment and special assessment;standardized processes and systematic inspection programs;examples of CUS applications;CUS training and the application of qualification certification.The establishment of applied technology standard is helpful for standardized training and clinical correct implementation.It is helpful for clinical evaluation and correct guidance treatment,and is also helpful for quality control and continuous improvement of CUS application.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.