To obtain active hFKBP52 protein for screening novel neu rotrophic drugs. Semi-nested and overlap PCR and affinity chromatography were u sed. hFKBP52 gene was cloned successfully from human fetal brain cDNA library, a nd then highly expressed (about 30%) as fusion protein in pET28a(+) vector syste m. The recombinant protein was purified as one band on SDS-PAGE. The purified h FKBP52 showed peptidyl-prolyl cis-trans isomerase (PPIase) activity, simil ar to the wild type.

Objective: Immunotoxin rRTA:DS27, which was prepared by conjugating DS27 with recombinanl ricin a chain, was compared with ricin:DS27 as immunotoxins. Methods: System analysis were performed regard to their cell-specific cytotoxicity, inhibition on the proliferation of hemoapeutic potential cells, immunogold-labelled intracellular routing and effect of NH_(4)Cl on the cytotoxicity. Results: Results showed that rRTA: DS27 got a more specific cytotoxicity and a weaker inhibition on the proliferation of hemoapeutic potential cells than ricin:DS27, NH_(4)Cl could obsolutelyy enhence the cytotoxicity of rRTA:DS27. Conclusion: rRTA:DS27 had more advantages than ricin: DS27 as immunotoxins.

Objective To evaluate the value of two-dimensional speckle tracking echocardiography (2D-STE) in assessing left ventricular systolic function,diastolic function and synchrony with different QRS complex duration in complete left bundle branch block (CLBBB) patients with preserved left ventricle ejection fraction (LVEF).Methods A total of 44 patients with CLBBB and LVEF≥50％ were included.All the patients were divided into two groups based on QRS duration,QRS＞150 ms as wide QRS group and 120 ms≤QRS≤150 ms as narrow QRS group.And 30 healthy people were included as control group.Two-dimensional echocardiography and 2D-STE were performed.Ieft ventricle longitudinal peak stain of global,septum and free wall (LS-G,LS-Sept,LS-Lat),standard deviation of time to peak systolic strain for the 18 left ventricular segments (SDt) and index of left yen tricular diastolic function (EDT,E/A and E/e') were measured.Results SDt values of wide QRS group and narrow QRS group were significantly higher than that of control group (both P＜0.01).And SDt of wide QRS group was significantly higher than that of narrow QRS group (P＜0.05).LVEF and LS-G in wide QRS group were significantly lower than those in both narrow QRS group and control group (all P＜ 0.05),while there was no significant difference between narrow QRS group and control group (all P＞0.05).The LS-Sept in wide QRS group and narrow QRS group were both lower than that of control group (both P ＜0.01).And LS-Sept in wide QRS group was lower than that of narrow QRS group (P＜0.01).LS-Lat in narrow QRS group was separately higher than those of both wide QRS group and control group (both P＜0.05),while there was no significant difference of LS-Lat between wide QRS group and control group (P＞0.05).Compared with control group,E/A and EDT decreased and of E/e' increased in both wide QRS group and narrow QRS group (all P＜0.05).While there was no significant difference between wide QRS group and narrow QRS group (all P＞0.05).Conclusion In patients of wide QRS CLBBB with preserved LVEF,left ventricular systolic,diastolic function and synchrony decrease,while left ventricular systolic function of patients with narrow QRS do not significantly decrease.

Objective:To assess the value of the ROX index in evaluating the efficacy of high-flow nasal cannula oxygen therapy (HFNC) in patients with coronavirus infected disease (COVID-19).Methods:This is a retrospective study. The included patients were diagnosed as COVID-19 in the intensive care unit (ICU) of the Cancer Center of Union Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from February 15, 2020 to March 15, 2020. All the patients were treated by HFNC. According to whether the patient subsequently received non-invasive positive pressure ventilation or invasive positive pressure ventilation, patients were divided into the HFNC success group and the HFNC failure group. Parameters in the two groups such as basic characteristics, lactic acid, number of chest radiographs, APACHE II, lymphocyte count, baseline respiratory rate, baseline percutaneous oxygen saturation, baseline PaO 2/FiO 2, baseline ROX index, and ROX index after 2, 6 and 12 h HFNC treatment were analyzed with t test, Chi-square test or rank sum test. Results:A total of 57 cases were included in this study. There were no significant differences in sex, age, comorbidities, lactic acid, quadrants of chest radiograph lung infection, APACHE II, lymphocyte count, and baseline respiratory frequency, transcutaneous oxygen saturation, oxygenation index, and ROX index between the HFNC success group and the HFNC faliure group ( P>0.05). Logistic regression analysis showed that ROX index after 2 h HFNC treatment ( OR=0.069), ROX index after 6 h HFNC treatment ( OR=0.194) and ROX index after 12 h HFNC treatment ( OR=0.036) were all protective factors for the therapeutic effect of HFNC treatment in COVID-19 patients. ROC curve showed that there were significant differences in ROX index after 2 h HFNC treatment, ROX index after 6 h HFNC treatment, and ROX index after 12 h HFNC treatment ( P<0.05). In the evaluation index, the area under the ROC curve of the ROX index after 2 h HFNC treatment was 0.838, the sensitivity was 64.5%, and the specificity was 100%. After 6 h HFNC treatment, the area under the ROX index ROC curve was 0.762, the sensitivity was 64.5%, and the specificity was 92.3%. After 12 h HFNC treatment, the ROX index ROC curve area was 0.866, the sensitivity was 67.7%, and the specificity was 100%. Conclusions:The ROX index can be used to evaluate the efficacy of HFNC in COVID-19 patients in a timely, simple and real-time manner.

The key to killing target cells by immunotoxin depends on the specific recognition of antibody to target cell and the cytotoxic effect of toxin. The comparative study of the killing effects of two anti-T immunotoxins, CD5:Ricin and CD5:rRA, on target cells was performed. The elimination rate of immunotoxins was analysed by flow cytometry and MLR. The effect of immunotoxins on the proliferation of hematopoiesis was evaluted by CFU-GM. The results showed that (1) CD5(+) T cells were eliminated and CD25(+) CD3(+) activated T cells were concentration-dependently inhibited by the two immunotoxins in the range of 10(-9) - 10(-11) mol/L; (2) both immunotoxins significantly inhibited the mixed lymphocyte reaction, and the inhibiting effect of CD5:rRA to T cell proliferation was markedly lower than that of CD5:Ricin in the range of 10(-10) - 10(-11) mol/L; (3) the combination of CD5:rRA with 10 mmol/L NH(4)Cl increased the T cell elimination rate; and (4) the two immunotoxins and the combination of NH(4)Cl and CD5:rRA did not suppressed proliferation of granulocyte-macrophage progenitors in the range of concentrations with killing effect. It was concluded that T cell and activated T cell could be eliminated effectively by immunotoxins, the proliferation of granulocyte-macrophage progenitor was not inhibited significantly.

Objective To evaluate left ventricular systolic and diastolic functions of patients with coronary slow flow (CSF) according to left ventricular myocardial systolic/diastolic performances (MSP/MDP) by using two-dimensional speckle tracking echocardiography (STE).Methods Fifty patients with CSF diagnosed with coronary angiography (CSF group) and 45 patients without CSF (control group) underwent STE.Left ventricular systolic longitudinal,radial and circumferential peak strain and early-diastolic peak strain rate were measured,and left ventricular MSP and MDP were calculated,then the results were statistically analyzed between the 2 groups.Results Compared with control group,left ventricular systolic longitudinal,radial and circumferential peak strain and early-diastolic peak strain rate,MSP and MDP reduced in CSF group (all P＜0.05).The mean thrombolysis in myocardial infarction (TIMI) frame count (TFC) of coronary artery was negatively correlated with MDP (r=-0.23,P=0.04),and the number of affected coronary arteries was negatively correlated with MDP (r=-0.31,P=0.03).There was significant difference of MDP among patients with different numbers of affected coronary arteries and control group (all P＜0.05),and MDP in affected 2 and 3 coronary arteries patients were lower than those of the control group (all P＜ 0.05).Conclusion Left ventricular systolic and diastolic functions is impaired in patients with CSF.The mean TFC and the number of affected coronary arteries are negatively correlated with left ventricular diastolic function.Left ventricular MSP and MDP are comprehensive parameters in evaluating systolic and diastolic functions.

Objective To evaluate the role of treadmill exercise‐stress echocardiography in the assessment of left ventricular ( LV ) function in patients with slow coronary flow ( SCF ) ,and discuss the possible mechanism of SCF . Methods Forty‐six patients with diagnosis of SCF were enrolled as SCF group . Forty age and sex matched adults were included as control group . All subjects had underwent clinical evaluation and exercise stress echocardiography . LV diastolic and systolic functions were assessed by conventional echocardiography , tissue Doppler imaging and two‐dimensional speckle tracking echocardiography at rest and during exercise . Results ①LV function in patients with SCF at rest :LV peak systolic longitudinal strain ( LS) was lower in patients with SCF than that in controls ( P ＜0 .05) . T here was no difference of LV ejection fraction ( LVEF) between the two group( P＞0 .05) . Early diastolic mitral annulus velocity ( M itral e′) and the ratio between the early mitral inflow velocity and Mitral e′( M itral E/e′) were significantly decreased in SCF group( all P＜0 .01 ) . ②LV function during exercise :LVEF ,LS and M itral e′were significant increased in two groups ( all P ＜0 .05 ) than those at rest ,but there was no significant difference of LVEF ,LS ,M itral e′ and M itral E/e′ between the two groups ( all P ＞ 0 .05 ) . Compared with control group ,ΔLS and ΔM itral e′were significantly higher in SCF group ( all P＜0 .05 ) . Conclusions LV systolic and diastolic function in patients with SCF are impaired at rest . LV systolic and diastolic function recover in patients with SCF during exercise .

Objective To evaluate the effect of septal flash (SF) on left ventricular function and systolic synchrony in patients with complete left bundle branch block (CLBBB) and preserved left ventricular ejection fraction (LVEF) by two-dimensional speckle-tracking imaging (2D-STI).Methods Sixty CLBBB patients with preserved LVEF were selected,and 30 healthy individuals serving as control group.The SF of CLBBB patients were evaluated using 2D-STI,and the CLBBB patients were categorized into two groups (SF group and NSF group) based on the present SF.Conventional echocadiography and 2D-STI were performed for all individuals.Results ①SF was present in 53.3％ of CLBBB patients with preserved LVEF and was not present in control group.②LV longitudinal peak stain of global (LS-G) in SF group was lower than those in both control group and NSF group (P ＜0.05),and there was no significant difference between control group and NSF group (P ＞0.05).③Compared with control group,there were significant increase of left atrial dimension and Mitral E/e'and decrease in mitral E/A and mitral e'in SF group and NSF group (P ＜0.05),while there was no significant difference between SF group and NSF group (P ＞0.05).④There was an increase in standard deviation of time to peak systolic strain for the 18 LV segments (LV-SDt) in SF group and NSF group,and the LV-SDt in SF group was higher than that in NSF group (P ＜0.01).Conclusions SF is present in 53.3 ％ of CLBBB patients,patients of CLBBB with SF have lower left ventricular systolic function and systolic synchrony.

OBJECTIVE To reveal the role of the basal forebrain(BF)GABAergic neurons in the regulation of isoflurane anesthesia and to elucidate the underlying neural pathways.METHODS The activity of BF GABAer-gic neurons was monitored during isoflurane anesthesia using a genetically encoded calcium indicator in Vgat-Cre mice of both sexes.The activity of BF GABAer-gic neurons was manipulated by chemogenetic and opto-genetic approaches.Sensitivity,induction time and emer-gence time of isoflurane anesthesia were estimated by righting reflex.The electroencephalogram(EEG)power and burst-suppression were monitored by EEG recording.The effects of activation of GABAergic BF-thalamic reticu-lar nucleus(TRN)pathway on isoflurane anesthesia were investigated with optogenetics.RESULTS The activity of BF GABAergic neurons was generally inhibited during isoflurane anesthesia,obviously decreased during the induction of anesthesia and gradually restored during the emergence from anesthesia.Activation of BF GABAergic neurons with chemogenetics and optogenetics promoted behavioral emergence from isoflurane anesthesia,with decreased sensitivity to isoflurane,delayed induction and accelerated emergence from isoflurane anesthesia.Optogenetic activation of BF GABAergic neurons prom-oted cortical activity during isoflurane anesthesia,with decreased EEG delta power and burst suppression ratio during 0.8%and 1.4%isoflurane anesthesia,respectively.Similar to the effects of activating BF GABAergic cell bod-ies,photostimulation of BF GABAergic terminals in the TRN also strongly promoted cortical activation and behav-ioral emergence from isoflurane anesthesia.CONCLU-SION The GABAergic neurons in the BF is a key neural substrate for general anesthesia regulation that facilitates behavioral and cortical emergence from general anesthe-sia via the BF-TRN pathway.