In this study, we constructed a yeast consortium surface-display expression system by using Flo1 as an anchor protein. Endoglucanase II (EGII) and cellobiohydrolase II (CBHII) from Trichoderma reesei, and β3-glucosidase 1 (BGLI) from Aspergillus aculeatus were immobilized on Saccharomyces cerevisiae Y5. We constructed the cellulose-displaying expression yeast consortium (Y5/fEGII:Y5/fCBHII:Y5/fBGLI = 1:1:1) and investigated the enzymatic ability and ethanol fermentation. The displayed cellulolytic enzymes was stabile during the 96-h fermentation. The yeast consortium produced 0.77 g/L ethanol from 10 g/L phosphoric acid swollen cellulose (PASC) within 96 h. The yield (in grams of ethanol produced per gram of carbohydrate consumed) was 0.35 g/g, which correspond to 68.6% of the theoretical yield.
Aspergillus ; enzymology ; Cellulase ; genetics ; Cellulose ; metabolism ; Cellulose 1,4-beta-Cellobiosidase ; genetics ; Enzymes, Immobilized ; genetics ; Ethanol ; metabolism ; Fermentation ; Glucosidases ; genetics ; Mannose-Binding Lectins ; metabolism ; Protein Binding ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; metabolism ; Trichoderma ; enzymology

Objective To establish a culture system in vitro of adult human retinal neural cells for providing a model for the research of retinal neural cells. Design Experimental study. Participants Cultured adult human retinal neural cells. Methods The isolated cells from adult human postmortem retina (20?40 years old) were cultured, then cells of different stages were identified with immunocytochemical staining and judged with phase contrast microscopy and electron microscopy. Main Outcome Measures Cellular morphology and structure. Results (1) The results of cell culture: the adult retinal neural cells could survive in vitro under some conditions and were identified as NSE positive mostly. (2) The results of electron microscopy: most cultured cells were photoreceptors, bipolar cells, horizontal cells and some were glial cells with scanning electron microscopy. Conclusions Under feasible conditions, the adult human retinal neural cells could be cultured and maintained effectively in vitro.

Objective To investigate the value and diagnostic efficiency of the quantitative dynamic contrast enhanced-magnetic resonance imaging (DCE-MRI) and intravoxel incoherent motion (IVIM) parameters using three dimention (3D)-histogram analysis for discriminating the Gleason score (GS) of prostate cancer. Methods A total of 53 patients pathologically confirmed as prostate cancer by systemic prostate biopsy who had routine , DCE and DWI-MRI scans were retrospectively analyzed. There were 15 cases for low-risk and 38 cases for intermediate/high-risk prostate cancer. The 3D ROI of all lesions based on T2WI was achieved by image registration to get the quantitative parameters of DCE-MRI and DWI-IVIM. The parameters of DCE-MRI contains: transfer constant (Ktrans), rate constant (Kep) and extracellular-extravascular volume fraction (Ve).The DWI-IVIM related quantitative parameters were ADC, diffusion coefficient (D), diffusion coefficient related to perfusion (D*) and perfusion fraction (f). Then the histogram analysis of these quantitative parameters was performed to get the mean, median, 25th percentile, 75th percentile, Skewness and Kurtosis. Using the Spearman rank correlation analysis to evaluate the correlation of these parameters and GS of prostate cancer. The diagnostic performance of these quantitative histogram parameters related to the GS in identifying low-risk and intermediate/high-risk of prostate cancer was carried by ROC. Results The Kep and Ktrans (mean, median, 25th, 75th) of DCE-MRI were positively correlated with GS (r value was 0.346 to 0.696, P<0.05). The ADC (mean, median, 25th, 75th), D (mean, median, 25th, 75th, Skewness, Kurtosis) and D*(25th) of DWI-IVIM were correlated with GS (r value was-0.544 to 0.428, P<0.05). The DCE-MRI quantitative parameters Kep (25th) had the highest area under curve (AUC, 0.961); The ADC (median) and D (25th) had higher AUC( 0.832, 0.888) in the quantitative parameters of DWI-IVIM, the difference between Kep(25th) and ADC (median) was statistically significant (Z value was 2.212, P value was 0.027). The difference of AUC between Kep (25th) and D (25th), D (25th) and ADC (median) was not statistically significant (Z values were 1.027 and 1.398, P values were 0.162 and 0.304, respectively).Conclusion DCE and IVIM quantitative parameters (Kep, Ktrans, ADC, D) histogram analysis results are correlated with GS, and can be used for distinguishing low-risk from intermediate/high-risk prostate cancer.

Objective: To analyze the infection composition and drug resistance to Gram-negative (G^-) bacilli in children′s respiratory tract in Suzhou, in order to provide evidence for rational use of antibiotics clinically. Methods: G^- bacilli culture samples were collected from 21 561 cases of nasopharyngeal secretions from patients with respiratory tract infection admitted at the Department of Respiratory, Children′s Hospital of Soochow University from January 2007 to December 2016, including 21 246 cases in general wards, and 315 patients who were transferred to the respiratory department after treatment in the Intensive Care Unit(ICU), and the children were divided into the general ward group and the ICU group, and the pathogens were compared and the changes in bacterial susceptibility were dynamically observed between the 2 groups. Results: The primary G^-bacteria for respiratory infection was Haemophilus influenzae, followed by Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii.The detection rates of Klebsiella pneumoniae and Pseudomonas aeruginosa in the ICU group were 16.8% (21/125 strains)and 14.4%(18/125 strains), respectively, which were significantly higher than those in the general ward group [10.0%(208/2 071 strains), 9.2%(190/2 071 strains)]. The detection rates of G^-bacteria in the ICU group were 33.7%(106/315 cases), which were significantly higher than those in the general ward group [9.4%(1 997/21 246 cases)], and the difference was statistically significant(χ²=210.325, P<0.001). The rare G^-bacillus such as Stenotrophomonas maltophilia, Acinetobacter junii and Burkholderia onion were higher in the ICU group [17.6%(22/125 strains)] than that in the general ward group [6.4% (132/2 071 strains)]. The rate that of G^-bacteria with two or more mixed infection in ICU group [17.0% (18/106 cases)] was significantly higher than in the general ward group [3.4%(68/1 997 cases)], and the difference was statistically significant(χ²=47.3, P<0.05). For the mixed infection, the ICU group was mainly composed of Klebsiella pneumoniae mixed with Pseudomonas aeruginosa or Escherichia coli, while the general ward group was composed of Haemophilus influenzae mixed with Pseudomonas aeruginosa or Escherichia coli.The sensitivity of Haemophilus influenzae to Ampicillin, Sultamicillin, Cefuroxime, Cefaclor and Azithromycin decreased, and the sensitivity to Chloramphenicol, Tetracycline and Trimethoprim+ Sulfamethoxazole increased year by year, and there were statistically significant differences in different years (all P<0.05). The sensiti-vity to Escherichia coli to Ceftazidime decreased year by year, and the sensitivity to Ampicillin and Levofloxacin increased year by year, and there were statistically significant differences in different years (all P<0.05). The sensitivity to Klebsiella pneumoniae to Cefoperazone/Sulbactam and imipenem decreased, and the sensitivity to Ciprofloxacin and Levofloxacin increased, and there were statistically significant differences in different years (all P<0.05). The sensitivity to Pseudomonas aeruginosa, Cefoperazone/Sulbactam and Ceftriaxone decreased year by year, and the sensitivity to Levofloxacin increased, and there were statistically significant differences in different years (all P<0.05). The detection rate of carbapenem-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa showed an increasing trend, and there were statistically significant differences in different years (all P<0.05). Conclusions The primary G^-bacteria for respiratory infections is Haemophilus influenzae, G^-bacilli especially, the mixed infection of G^-bacilli, is more likely to cause severe and critical respiratory infections.The resistance rate of G^-bacteria infection in children′s respiratory tract to commonly used antibiotics is generally on the rise.

Objective:To analyze the relationship between plasma fibrinogen(FIB) within normal range and microalbuminuria in elderly patients with type 2 diabetes mellitus.Methods:A total of 869 elderly subjects with type 2 diabetes mellitus admitted to the Department of Endocrinology of Shanghai Fifth People′s Hospital from October 2012 to October 2014 were included in the study. The patients were divided into four groups based on the quartile level of FIB: Q1 group(<2.42 g/L), Q2 group(2.42-2.89 g/L), Q3 group(2.90-3.61 g/L), and Q4 group(≥3.62 g/L). The relationship between FIB and urinary albumin/creatinine ratio(UACR) was analyzed.Results:With the increasing of FIB, the level of UACR was significantly elevated( P<0.05). Pearson correlation analysis showed that FIB was positively associated with age, duration of diabetes, creatinine(Cr) and UACR in men and women( P<0.01). Multiple regression analysis showed that FIB was an independent factor of UACR( P<0.01). Logistic regression analysis showed that the risks of microalbuminuria and macroalbuminuria were respectively 4.536 folds(95% CI 2.516-8.175, P<0.01) and 13.314 folds(95% CI 2.925-60.612, P<0.01) in Q4 group, and 2.177 folds(95% CI 1.273-3.724, P<0.01) and 4.098 folds(95% CI 1.101-19.226, P<0.05) in Q3 group as compared with Q1 group after adjused by following factors: gender, age, duration of diabetes, body mass index(BMI), systolic blood pressure(SBP), diastolic blood pressure(DBP), fasting plasma glucose(FPG), HbA 1C, total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), Cr, alanine aminotransferase(ALT), as well as smoking and drinking behavior. Based on the cut off values to UACR 30 mg/g and 300 mg/g, the receiver operating characteristic curve(ROC) was used to evaluate the value of FIB for UACR. The optimal cut-off value of FIB was 3.18 g/L and 3.22 g/L respectively. Conclusions:Plasma FIB was closely associated with microalbuminuria in elderly patients with type 2 diabetes mellitus, which may be considered as one of the predictors for diabetic nephropathy.

Objective:To explore the consistency of MRI-based ovarian-adnexal report and data system (O-RADS) score and its diagnostic value for ovarian adnexal masses.Methods:The MRI data of 309 patients with ovarian adnexal masses confirmed by pathology were retrospectively collected from January 2017 to August 2021 in the Second Affiliated Hospital of Soochow University, including 327 lesions consisted of 250 benign lesions, 21 borderline lesions, and 56 malignant lesions confirmed by pathology. Borderline and malignant lesions were classified into the malignant group ( n=77) and benign lesions were classified as benign group ( n=250). Two radiologists scored all lesions according to the MRI-based O-RADS, and scored again after 6 months. The proportion of borderline/malignant lesions in each MRI-based O-RADS score was calculated. The weighted Kappa test was used to assess the intra-reader and inter-reader consistency of the image interpretation results. The receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic efficacy of MRI-based O-RADS classification for distinguishing benign and malignant ovarian adnexal masses. Results:The weighted Kappa value of the MRI-based O-RADS score between the two radiologists was 0.810 (95%CI 0.764-0.855), and the weighted Kappa values of the two radiologists′ scores at different times were 0.848 (95%CI 0.806-0.889) and 0.875 (95%CI 0.835-0.914), respectively. The borderline/malignant lesions accounted for 0/16, 0.8% (1/127), 10.1% (10/99), 76.0% (57/75), 9/10 and 0/17, 0 (0/122), 8.0% (8/100), 76.2% (48/63), and 84.0% (21/25) of the lesions in the two radiologists based on the MRI O-RADS score of 1, 2, 3, 4, and 5, respectively. When adopting O-RADS score>3 as a cut-off value, the area under the ROC curve of the two radiologists for distinguishing benign and malignant ovarian adnexal masses was 0.928 (95%CI 0.895-0.954) and 0.942 (95%CI 0.911-0.965), respectively. The sensitivity was 0.857 and 0.896, the specificity was 0.924 and 0.924, and the accuracy was 0.908 and 0.917 respectively.Conclusion:The MRI-based O-RADS yields high diagnostic efficiency in the evaluation of benign and malignant ovarian adnexal masses, and the intra-reader and inter-reader consistency of the image interpretation is strong.

Objective To explore the difference in efficacy between multiparametric MRI (Mp-MRI) based on prostate imaging reporting and data system version 2 (PI-RADS v2) and abbreviated biparametric MRI (Bp-MRI) in detecting prostate cancer (PCa) and clinically significant prostate cancer (csPCa), and to evaluate the consistency of image interpretation between different readers. Methods The imaging, pathological and clinical data of patients with prostatic Mp-MRI in our hospital from February 2015 to June 2018 were retrospectively analyzed. At the beginning, 250 patients were randomly selected. Two radiologists visually evaluated the images of those patients using two 5-point scoring schemes based on Mp-MRI and Bp-MRI. The remaining cases were independently proceeded by one of the radiologists using two schemes respectively. Weighted Kappa test was used to assess the consistency of the results interpreted by the two radiologists. The receiver operating characteristic (ROC) curve was used to evaluate the efficiency of the two scoring schemes in detecting PCa and csPCa, and with Z test to investigate whether there was any difference in detection efficiency between the two schemes. Results Nine hundred and seventy eight patients were eventually enrolled in the study. The results of the consistency assessment showed that there was good agreement between the two radiologists, whether using Mp-MRI or Bp-MRI, with the weighted Kappa coefficient of 0.800 and 0.812, respectively. The ROC curve analysis showed that the area under the curve (AUC) of PCa detected by Mp-MRI and Bp-MRI was 0.873 and 0.879, respectively, and the AUC of csPCa detected was 0.922 and 0.932, respectively. In addition, there was no statistically significant difference between the AUC of PCa and csPCa detected by the two schemes (P>0.05). Conclusion The Bp-MRI scoring scheme has good stability in the evaluation of benign and malignant prostate, and its detection efficiency of PCa or csPCa is not lower than that of standard Mp-MRI based on PI-RADS v2.

OBJECTIVE To evaluate th e effects of comprehensive drug cost control in China ，and to provide reference for further improving the effects of cost control. METHODS Entropy method was used to establish a comprehensive evaluation index system of the comprehensive drug cost control effect from the respective of drug price control ability ，drug cost control ability and patient affordability. The comprehensively evaluate the effects of drug cost control in 31 provinces （autonomous regions and municipalities）during 2016-2020. The k-means cluster method was used to analyze the effects of comprehensive drug cost control in various provinces. RESULTS & CONCLUSIONS During the period of 2016-2020，the total score of comprehensive drug cost control effect of 31 provinces were 14.64，16.71，17.58，17.57，17.88，respectively. The results of cluster analysis were similar to the ranking of entropy method. Medical and health system reform policy had entered a stable period after achieving phased results ； the effects of comprehensive drug cost control was characterized by regional steps in 31 provinces；the effects of comprehensive drug cost control were better in developed coastal areas and some provinces and cities of western regions ，followed by provinces and cities in central China ；however，comprehensive drug cost control in 3 provinces of northeast China showed poor effect. The effects of comprehensive drug cost control in pilot cities of comprehensive medical reform were significantly improved. It is suggested that the successful experience of pilot cities of comprehensive medical reform should be promoted nationwide ，and policies such as volume-based drug procurement ，medical insurance drug price negotiation ，and diagnosis-related groups / diagnosis-intervention packet payment method reform should be further deepened.