Zearalenone is one of the most widely polluted Fusarium toxins in the world, seriously endangering livestock and human health. Zearalenone hydrolase (ZHD) derived from Clonostachys rosea can effectively degrade zearalenone. However, the high temperature environment in feed processing hampers the application of this enzyme. Structure-based rational design may provide guidance for engineering the thermal stability of enzymes. In this paper, we used the multiple structure alignment (MSTA) to screen the structural flexibility regions of ZHD. Subsequently, a candidate mutation library was constructed by sequence conservation scoring and conformational free energy calculation, from which 9 single point mutations based on residues 136 and 220 were obtained. The experiments showed that the thermal melting temperature (T_m) of the 9 mutants increased by 0.4-5.6 ℃. The S220R and S220W mutants showed the best thermal stability, the T_m of which increased by 5.6 ℃ and 4.0 ℃ compared to that of the wild type. Moreover, the thermal half-inactivation time at 45 ℃ were 15.4 times and 3.1 times longer, and the relative activities were 70.6% and 57.3% of the wild type. Molecular dynamics simulation analysis showed that the interaction force at and around the mutation site was enhanced, contributing to the improved thermal stability of ZHD. The probability of 220-K130 hydrogen bond of the mutants S220R and S220W increased by 37.1% and 19.3%, and the probability of K130-D223 salt bridge increased by 30.1% and 12.5%, respectively. This work demonstrated the feasibility of thermal stability engineering strategy where the structural and sequence alignment as well as free energy calculation of natural enzymes were integrated, and obtained ZHD variants with enhanced thermal stability, which may facilitate the industrial application of ZHD.
Humans ; Hydrolases ; Zearalenone ; Trichothecenes ; Gene Library ; Hydrogen Bonding

Objective:To explore the efficacy and safety of intraarterial microguidewire electrocoagulation in arterial aneurysms.Methods:(1) SilverSpeed, a kind of microguidewire used in clinical intravascular treatment for intracranial aneurysms, was used to conduct in vitro electrolysis gas generation experiment with isolated arterial blood of anticoagulant New Zealand white rabbits as medium, and thrombus attachment on the surface of microguidewire was observed under scanning electron microscope. (2) Rabbit common carotid artery aneurysm models were established by using vein bag transplantation method, and divided into microguidewire electrocoagulation treatment groups ( n=40) and blank control group ( n=10). The number of closured tumor cavity and the quality of formed thrombus were observed after electrocoagulation simulation treatment with SilverSpeed microguidewire (charging at 6, 9, 12, 15, and 18 V voltage, respectively for 1, 3, 6, 9, 12, and 15 min). DSA was used to observe whether there was ruptured aneurysms or thrombosis of parent artery. Twelve h later, head MRI diffusion weighted sequence scan was performed to detect whether there were new cerebral ischemia foci in the distal cerebral blood supply area of the parent artery. DSA was performed again 6 months after surgery to observe whether the aneurysms recurred. Results:(1) Electrolytic gas generation experiment results showed that bubbles were generated after electrification of SilverSpeed microguidewire; the higher the voltage, the more severe the reaction. Scanning electron microscope showed that thrombus attached to the surface of the microguidewire after electrification in isolated blood; and the higher the voltage, the denser the thrombus. (2) Under the same charging time, the higher the voltage, the larger the number of closured tumor cavity in rabbits of the microguidewire electrocoagulation treatment groups. Under the same voltage, the longer the charging time, the better the quality of thrombosis. Ischemic events occurred only in the microguidewire electrocoagulation treatment group with voltage>9 V, and the charging duration was not associated with the incidence of embolic events. When the voltage was 15 V, 2 experimental rabbits died due to aneurysm rupture 3 min after electrification. When the voltage was 18 V, 4 experimental rabbits died of cardiac arrest 9 min after electrification, and another 2 rabbits died of aneurysm rupture 6 min after electrification.Conclusions:High voltage is the main cause of adverse events in the microguidewire electrocoagulation treatment of aneurysms. After setting the appropriate voltage, prolonging the electrification time can improve the electrocoagulation effect without increasing the safety risk.

OBJECTIVE：To provide reference for improving the level of pediatric pharmacoeconomic research in China. METHODS： Using “Pediatric”“Pharmacoeconomics”“Cost-effectiveness”“Cost-utility”“Cost-benefit”“Minimum cost ” as keywords，pediatric pharmacoeconomic research literatures published during Jan. 2009 to Dec. 2018 were retrieved from CNKI ， Wanfang，VIP and CBMdisc. The Pediatric Quality Appraisal Questionnaire was used to evaluate the quality of the literatures. The problems of these literatures were analyzed to put forward the suggestions. RESULTS & CONCLUSIONS ：Totally 140 literatures were eventually included. The number of published literatures was increasing and the distrubtion of journals was scattered. The first auther was mainly from medical institutions. Included literatures mainly involved respiratory disease and drug treatment ，etc. Among them ，15 literatures indicated the research angle ；research time limit of 109 literatures was equal to or less than one year ；6 literatures carried out sensitivity analysis. Most of them adopted cost-effectiveness analysis and decision analysis model. The results of quality evaluation showed that the economic evaluation ，camparators，target population ，outcomes，analysis and conclusions domains had high scores （0.62 to 1.00 score），time horizon and discounting domains had middle scroes （0.49 and 0.53 score respectively），but perspective ，costs and resource use ，incremental analysis ，sensitivity analysis and conflict of intrest domains had low scores （0.10 to 0.31 score）. The main problems of pediatric pharmacoeconomic literatures in China focused on selection of research perspective ，measurement of costs and clinical outcomes ，and selection of analytical methods. The quality of pediatric pharmacoeconomic evaluations in China need to improve. It is suggested to supplement the content of pediatric pharmacoeconomics in China Guidelines for Pharmacoeconomic Evaluations . It is also suggested for pediatric pharmacoeconomic evaluations to clarify the perspective ，standardize the cost identification ，reasonably select health outcome indicators and correctly apply analysis method in order to improve the quality.

Objective To investigate the prediction of axillary lymph node metastasis in patients with early breast cancer by sentinel lymph node biopsy (SLNB) combined with preoperative ultrasound.Methods From Jun 2014 to Oct 2018,340 patients with early breast cancer were enrolled in the 5th Department of General Surgery of Second Hospital of Lanzhou University.113 cases were grouped into ultrasound examing,75 patients in the SLNB,and 152 patients in the SLNB combined with preoperative ultrasound.Using the results of axillary lymph node dissection as gold standard,the 3 groups were compared.Results The sensitivity of SLNB,ultrasound and SLNB combined with ultrasound for suspicious axillary lymph node biopsy were 90.1％,85.7％,and 96.4％,respectively.The specificity was 84.0％,76.3％,and 100％,and the accuracy was 91.6％,83.0％,and 97.4％,respectively.The false negative rates were 8.6％,14.3％,and 3.6％,respectively.Conclusion Preoperative ultrasound diagnosis of axillary lymph node status combined with sentinel lymph node biopsy can significantly improve the diagnosis rate of axillary lymph node metastasis in early breast cancer.

OBJECTIVE: Patients with chronic neuropathic pain (CNP) have a higher incidence to develop depression. However, its pathogenesis has not yet been fully elucidated. Here we aimed to investigate the role of inflammatory cytokines in CNP-related anhedonia, which is a core symptom of depression, and to explore the effects of ketamine and parecoxib on pain and anhedonia. METHODS: A rat model of spared nerve injury (SNI) was constructed to mimic CNP. Hierarchical cluster analysis of sucrose preference test (SPT) was applied to classify the SNI rats into anhedonia susceptible and unsusceptible. Inflammatory cytokines in medial prefrontal cortex (mPFC) of brain, serum and L2–5 spinal cord were measured. Moreover, effects of ketamine or parecoxib on mechanical withdrawal test (MWT) and SPT in anhedonia susceptible rats were detected. RESULTS: Tumor necrosis factor (TNF)-α was increased in mPFC, serum and and spinal cord of anhedonia susceptible rats. Furthermore, anhedonia susceptible and unsusceptible rats both increased the interleukin (IL)-1β level in mPFC, serum and spinal cord. IL-6 was altered in serum and spinal cord, but not in mPFC. IL-10 was significantly altered in mPFC and serum, but not in spinal cord. Additionally, ketamine treatment significantly attenuated the decreased results of MWT and SPT in anhedonia susceptible rats, and that parecoxib significantly improved the MWT score, but failed to alter the result of SPT. CONCLUSION: These findings suggest that abnormalities in inflammatory cytokines confer susceptible to anhedonia in a rat model of SNI. Ketamine, a fast-acting antidepressant, has pharmacological benefits to alleviate pain and anhedonia symptoms.
Anhedonia ; Animals ; Brain ; Cytokines ; Depression ; Humans ; Incidence ; Interleukin-10 ; Interleukin-6 ; Interleukins ; Ketamine ; Models, Animal ; Neuralgia ; Neurogenic Inflammation ; Prefrontal Cortex ; Rats ; Spinal Cord ; Sucrose ; Tumor Necrosis Factor-alpha

OBJECTIVE： To explore the current situation of service ability and pharmaceutical service construction of primary healthcare institutions in Jiangsu province. METHODS： Questionnaire survey was conducted among primary healthcare institutions of 13 prefecture level cities in Jiangsu province， involving basic service information （such as serving mode， medical departments）， resource allocation （such as hardware， human resource） and pharmaceutical care construction （such as the leaders of the drug procurement， pharmaceutical departments allocation， mediation safety management）. The current situation of service ability and pharmaceutical service construction was evaluated in primary healthcare institutions of Jiangsu province. RESULTS： A total of 579 primary healthcare institutions in Jiangsu province were investigated and 579 questionnaires were collected （some items in the questionnaires were not answered）. Outpatient service （543/555，97.8％） was the main mode of service provided by primary healthcare institutions in Jiangsu province， followed by emergency call （321/555，57.8％） and hospitalization （292/555，52.6％）； the frequently established department in primary healthcare institutions was the medical department for all （452/568，79.6％）， preventive healthcare department（401/568，70.6％）， internal medicine department （365/568，64.3％）； 16.1％ （93/533）of the primary healthcare institutions had no beds； 36.4％ （211/579）of the institutions had no chief physician； 20.9％ （111/532）of the primary healthcare institutions medicine procurement were managed by pharmacists； 42.5％ （212/499）of the institutions had no pharmaceutical departments； physicians were responsible for patients' medication safety in 43.9％ （236/537） of the primary healthcare institutions and pharmacists were responsible for patients' medication safety in only 16.2％ （87/537）of the institutions. CONCLUSIONS： Primary healthcare institutions in Jiangsu province have made some achievements in service capability building. Human resources， equipment and facilities， pharmaceutical care construction and other aspects still need to be improved.

Objective To analyze the risk factors of acute kidney injury after orthotropic liver transplantation for adult benign end-stage liver disease.Methods A retrospective analysis was conducted in 30 recipients (18 males,12 females,aged 23-68 years,ASA grade Ⅲ or Ⅳ) who underwent orthotropic liver transplantation for benign end-stage liver disease at Tongji Hospital from May,2014 to December,2014.Both demographic characteristics and perioperative parameters were collected,including general condition,surgery and anesthesia factors and intraoperative salvage autotransfusion or not.Perioperative laboratory findings related to renal function including urine volume,serum creatinine (Scr) and urea nitrogen (BUN) were collected,too.All variables tested in the univariate analysis with a P<0.10 were included in a multiple logistic regression analysis.Results There were less intraoperative salvage autotransfusion,more platelet transfusion and a higher using rate of vasopressors in the AKI group after surgery than those did not.Patients who received intraoperative salvage autotransfusion had 0.058 time odds (95%CI 0.005-0.649) of AKI than those did not;patients who required platelet transfusion had 10.706 times higher odds (95%CI 1.212-94.963) of AKI than those did not.Conclusion It is likely that intraoperative salvage autotransfusion was able to decrease the morbidity of AKI,while platelet transfusion and vasopressor administration to maintain blood pressure could increase the possibility of AKI.

Objective To understand the situation of college students' awareness about innovative credit, analyze the influencing factors, in order to provide a scientific basis for constructing innovation credits and promoting the double strategy. Method The undergraduates of a medical university who were still at school in April 2015 were selected, and by using a stratified cluster sampling method, 450 students were chosen to be conducted by self-administered questionnaire survey to understand the cognitive status of in-novation of credit system in college students. Chi square test was used to compare the passing rates of students in different demographic characteristics, and the relationship between scores was analyzed by linear regression analysis. Result A total of 439 valid questionnaires were collected , and the total scores of innovative knowledge, attitudes and behaviors of 439 undergraduates were 15.95％ (70/439), 36.44％ (160/439) and 32.80％(144/439) respectively. In the part of knowledge innovation credits, girls' passing rate was higher than boys ( χ2=4.010,P=0.045. In the part of attitude, high grade group students had more positive attitude than the low grade group of students ( χ2=6.227,P=0.0013). In the part of behavior; higher grade students with innovative credits had higher pass rates than those in lower grades (chi, 2=7.781, P=0.005), and boys had a higher rate of passing than girls ( χ2=6.658,P=0.010). The total score of knowledge was positively linear with the total score of attitude and behavior. Conclusion College Students' awareness of innovation credit rate is low, but the innovative attitude and behavior is positive. The higher the awareness rate is conducive to the cultivation of attitude and behavior, so it is necessary in medical colleges to carry out innovation of credit and promote the cultivation of innovative consciousness.

Objective To systematically analyze and assess the overall value of transrectal shear wave elastograpy in diagnosis of prostate cancer (PCa) using Meta-analysis.Methods Relevant Chinese and foreign papers diagnosing PCa with transrectal shear wave elastograpy published before December 2016 were searched.The references were evaluated and screened according to the criteria of diagnostic research.The selected references were analyzed by Meta-Disc 1.4 and Stata 12.0 statistical software.Results Eight articles were included in the present Meta-analysis.Five of these literatures were about transrectal shear wave elastograpy in diagnosis of PCa,the summarized sensitivity (SEN) and summarized specificity (SPE) in diagnosis of PCa were 0.80 (95％CI [0.75,0.84]) and 0.75 (95％CI [0.71,0.79]),respectively;the positive likelihood ratios (PLR) and negative likelihood ratios (NLR) were 3.60 (95 ％ CI [2.57,5.05]) and 0.17 (95 ％ CI[0.08,0.37]),respectively;the area under SROC curves was 0.895.Five of these literatures were about transrectal shear wave elastograpy supplemental prostatic biopsy in diagnosis of PCa,the summarized SEN and SPE were 0.86 (95％ CI [0.83,0.88]) and 0.84 (95％CI [0.82,0.85]) respectively;the PLR and NLR were 5.81 (95％CI [3.07,10.99]) and 0.14 (95％CI [0.04,0.49]) respectively;the area under SROC curves were 0.924.Conclusion Transrectal shear wave elastograpy has better clinical value in detection of PCa and can be used to supplemental prostatic biopsy.

Objective To discuss the clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope and its surgical techniques. Methods Clinical data of 161 patients with chronic subdural hematoma from August 2009 to December 2015 was analyzed retrospectively. 74 of them experienced surgeries assisted by rigid neuroendoscope (endoscope group) and other 87 cases were operated without neuroendoscope (routine group) during the same period. Results Although there were significant difference in operative duration between the two groups, complications, ratio of total removal of hematoma after surgery, postoperative inpatient duration and recurrent rate of hematoma were more advantageous in endoscope group. The operative duration of endoscope group with (112.68 ± 34.86) min was longer than that of routine group with (74.11 ± 28.23) min (t = 7.75, P = 0.000), while the postoperative inpatient duration of endoscope group with (8.23 ± 2.01) d was shorter than that of another group with (10.79 ± 5.02) d (t = -4.12, P = 0.000). There were no surgical associated complications in endoscope group, but 1 patient in routine group experienced intracerebral hematoma of frontal lobe and associated aphemia. Total removal of hematoma was confirmed in endoscope group with 98.65% (73/74), which was higher than that in routine group with 86.21% (75/78) (χ2 = 8.34, P = 0.004). Hematoma recurrence was found in 16 cases of routine group (18.39%), but more superiority in endoscope group with 1.35% (χ2 = 12.29, P = 0.000). Outpatient follow-up was carried out in all patients from 6 to 38 months with an average duration of 30.06 months. In 17 cases with recurrent hematoma during follow-up, 15 of them were cured by a second surgery, and another 2 patients were cured by atorvastatin. Conclusion As a simple, safe and effective technique, the application of rigid neuroendoscope during surgery for chronic subdural hematoma is more advantage than routine surgery. A self-made suction with adjustable soft curved tip is suitable for such procedure.