β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.
beta-Glucosidase ; Sodium Chloride ; Temperature ; Glucose ; Ethanol/chemistry* ; Ions ; Hydrogen-Ion Concentration ; Enzyme Stability ; Substrate Specificity

Objective:To investigate the clinical features and influencing factors of long-term prognosis of tuberculous meningitis(TBM), and to provide a recommendation for treatment and early intervention of TBM.Methods:Clinical data of TBM patients were retrospectively collected at Peking Union Medical College Hospital from January 2014 to December 2021. Patients who were followed-up more than one year were divided into two groups according to modified Rankin Scale (mRS). Risk factors associated with long-term prognosis were analyze by conditional logistic stepwise regression.Results:A total of 60 subjects were enrolled including 33 (55%) males and 27 (45%) females with age 15-79 (44.5±19.8) years. There were 30 cases (50%) complicated with encephalitis, 21 cases (35%) with miliary tuberculosis. The diagnosis was microbiologically confirmed in 22 patients (36.7%), including 5 cases (22.7%, 5/22) by acid-fast staining, 8 cases (36.4%, 8/22) by Mycobacterium tuberculosis (MTB) culture, and 20 cases (90.9%, 20/22) by molecular biology. The median follow-up period was 52(43, 66 ) months in 55 cases surviving more than one year. Among them, 40 cases (72.7%) were in favorable group (mRS 0-2) and 15 cases (27.3%) were in unfavorable group (mRS 3-6) with poor prognosis. The mortality rate was 20% (11/55). Elderly ( OR=1.06, P=0.048 ) , hyponatremia( OR=0.81, P=0.020), high protein level in cerebrospinal fluid (CSF) ( OR=3.32, P=0.033), cerebral infarction( OR=10.50, P=0.040) and hydrocephalus( OR=8.51, P=0.049) were associated with poor prognosis in TBM patients. Conclusions:The mortality rate is high in patients with TBM. Molecular biology tests improves the sensitivity and shorten the diagnosis time of TBM. Elderly, hyponatremia, high protein level in CSF, cerebral infarction and hydrocephalus are independent risk factors of long-term survival in TBM patients.

Objective: To detect the hemodynamic mechanism of the novel endovascular stent on complicated abdominal aortic aneurysms by Computational Fluid Dynamics(CFD) firstly, and then compare the effect of the novel endovascular stent and the streamliner multilayer flow modulator(SMFM) stent. Methods: All medical images were obtained by computed tomography. A semiautomatic segmentation protocol within Mimics(v17.0; Materialise, Leuven, Belgium) was used to extract the threedimensional aortic aneurysms. The stents was generated numerically and fitted along the aortic aneurysms. The lumen volume represented the fluid domain that was discretised in smaller volumes, which defined a mesh within the ICEM software(Ansys ICEM CFD v15.0). Hemodynamic analysis was performed with software Fluent 16.0. Results: Both kinds of stents can change the pattern of flow distribution. Compared with SMFM, the novel endovascular stent can significantly reduce the flow velocity in aneurysms, the shear force and the pressure on the aneurysms wall.What’s more, the flow velocity of the branch artery was accelerated by the novel endovascular stent. Conclusion The novel endovascular stent can significantly reduce the flow velocity in aneurysms, the shear force and the pressure on the aneurysms wall, and acceleratethe the flow velocity of the branch artery.

Objective Big endothelin-1(big ET-1) is associated with the prognosis of various cardiovascular diseases. This study is to explore the predictive value of plasma big ET-1 level for long-term outcome after mitral valvuloplasty in patients with moderate to severe chronic ischemic mitral insufficiency .Methods A total of 142 consecutive patients with moderate to severe chronic ischemic mitral regurgitation who underwent coronary artery bypass grafting and mitral valvuloplasty were enrolled at our center from January 2009 to December 2015.The clinical baseline data were collected and recorded.All patients were followed up.The mortalities and major adverse cardiovascular and cerebrovascular events during the follow-up period were re-corded.Results A total of 142 patients were enrolled, with mean age of(59.4 ±8.3) years.During(51.9 ±22.6) months follow-up, 19 cases(13.4％) died and 35 cases(24.6％) had major adverse cardiovascular and cerebrovascular events.Uni-variate and multivariate Cox regression analysis showed that big ET-1 was an independent risk factor for all-cause mortality (HR=3.970, 95％CI:1.535-10.268) and major adverse cardiovascular and cerebrovascular events(HR=2.754, 95％CI:1.238-6.129).In addition, left ventricular ejection fraction, pulmonary hypertension and stroke history were independent risk factors for all-cause mortality , while left ventricular ejection fraction , stroke history were independent risk factors for major adverse cardiovascular and cerebrovascular events.Conclusion Long-term outcomes of simultaneous coronary artery bypass grafting and mitral valvuloplasty for moderate to severe ischemic mitral insufficiency are satisfactory and big ET-1 is an inde-pendent risk factor for long-term outcomes of these patients.

Fever and abdominal pain are common symptoms and could be main manifestations in patients with autoinflammatory diseases.A 48-year-old female patient was admitted with recurrent fever and abdominal pain for 9 years.Serum level of inflammatory markers synchronously fluctuated with fever,and returned to normal when fever subsided.The periodic episodes of fever occurred every 1 to 4 months and failed to respond to empirical antibiotics.Whole exome sequencing showed heterozygous mutation of NOD2 gene q902k,leading to the final diagnosis of autoinflammatory disease.Corticosteroid and tripterygiumglycosides were effective for the disease remission.

Objective To assess the clinical value of cross raft screws technique in preventing postoperative collapse of tibial plateau fracture.Methods From September 2014 to November 2017,data of 14 patients with tibial plateau fracture who were treated by cross raft screws technique were retrospectively analyzed.There were 9 males and 5 females aged from 30-65 years old (average,44.4± 1.7 years).There were 8 patients that the thickness of subchondral cancellous bone measured preoperatively by CT data was less than 4 mm,and 6 patients that the thickness of subchondral cancellous bone measured intraoperatively was less than 4 mm.1/4 tubular plate was placed along the anterior rim of lateral tibial plateau,and the 3.5 mm cortical bone screws were fixed as bamboo raft from anterior to posterior through the plate.The collapse of tibial plateau after surgery were measured by CT scan after union of the fracture.The function of knee was evaluated by Rasmussen Anatomical and Functional Grading.Results The time of tubular plate procedure was 18 to 35 min (average,24.1±5.4 min).All 14 patients were successfully followed-up for 13.8±5.1 months.The height of collapse preoperatively by CT scan was 5-21 mm (average,8.00± 1.40 mm).3 days after the operation,the height between articular line and lateral articular surface was 0-2 mm (average,0.80±0.06 mm).Compared with CT data preoperatively,the collapse was corrected postoperatively that was proved by CT scan (P＜ 0.05).After the fracture was healed,according to CT data,the height between articular line and lateral articular surface was 0-2 mm (average,0.70±0.08 mm).Compared with CT data postoperatively,there was no postoperative collapse happened (P=0.466).The position and length of nails were placed appropriately.The average healing time of fracture was 3.6 months.There were no infection,nonunion and pain of tendon happened.The plate could be touched subcutaneously in 2 patients,who had no discomfort feelings.The patient's postoperative Rasmussen Anatomical Grading were 13-18 (average,16.7),including 8 cases excellent and 6 cases good.The postoperative Rasmussen Functional Grading was 18-28 (average,25.7),including 11 cases excellent and 3 cases good.Conclusion The cross raft screws technique is a good way to prevent the postoperative collapse of the lateral articular surface of tibial plateau.

Objective To analyze the epidemiology and clinical characteristics of patients with Brucella endocarditis in order to improve the understanding and treatment of Brucella endocarditis.Methods The clinical data including demographic characteristics,clinical features,laboratory data,echocardiography,treatment and clinical outcome of 6 patients with Brucella endocarditis were collected and analyzed.Results In our database consisting of 211 brucellosis patients with positive blood culture,6 patients (2.8％) with Brucella endocarditis were selected from May 2007 to December 2016 in Peking Union Medical College Hospital.The age range was 25-55 years,among whom 5 patients were men.The majority were farmers and all patients reported the history of closely contact with livestock.All of them had fever and 3 of them had arthralgia.Four patients had aortic involvement (4/6) and 2 of them presented with symptoms of left ventricular failure at admission.Blood culture of Brucella and serum agglutination test of Brucella were positive in all of them.Combined antibiotics treatments were given to all patients.Valve replacement surgery was performed in four patients,among them,three patients well recovered,one lost.Among all the patients,four were followed up continually and other two were lost.Conclusions Brucella endocarditis has a predominance of aortic involvement and is prone to left ventricular failure.Early antibiotic treatment combined with valve-replacement surgery is effective to improve the prognosis.

Objective To study the feasibility and safety of coronary sinus (CS) ventricular pacing comparing with traditional right ventricular (RV) pacing in ordinary pacing treatment.Methods Sixty-one patients with an indication of pacemaker implantation were randomized into traditional RV pacing group and CS ventricular pacing group.The success rate,complications,preoperative and postoperative QRS wave duration and pacemaker parameters were compared between the two groups.Results (1) The instant success rate in traditional RV pacing and the CS pacing groups were 100％ and 68.97％ (P＜0.01);the surgical success rate was significantly higher by using left ventricular electrode than the ventricle tined electrode in CS pacing group (85.71％ and 25％,P＜0.05).There was no significant difference in the incidence of complications between the two groups (P＞0.05);(2) The increase of QRS complex width at postoperative 1 month in CS pacing group is significantly less than the RV pacing group (P＜0.05);(3) Although the pacing threshold and electrode impedance during operation,postoperative 1 month and 3 months in CS pacing group were higher than RV pacing group (P＜0.05),the CS pacing was effective.Conclusion The CS ventricular pacing is as safe and effective as traditional RV pacing,and is more consistent with physiological ventricular activation sequence.Using left ventricular electrode can significantly improve the success rate of CS ventricular pacing.

[Abstract ] Objective Cardiac shock wave therapy (CSWT) can promote arteriogenesis in ischemic myocardia , but the mo-lecular mechanism remains unclear .The study aimed to explore the effect of CSWT on arteriogenesis in human cardiac microvascular endothelial cells ( HCMEC ) and the role of focal adhesion kinase (FAK) and Calcium-activated potassium channels (KCa) in the sig-nal conduction pathway of CSWT arteriogenesis . Methods HC-MEC cells cultured in vitro were randomly divided into control group , CSWT group , CSWT +T ( FAK inhibitor PF-573228 ) group and CSWT+F( SCa inhibitor iberiotoxin ) group.Each group received one CSWT(0.09 mJ/mm2, 200Times) 48 h after added stimulant.24 hours'conventional culture later , tests were made on the levels of endothelial nitric oxide synthase ( eNOS ) and vascular endothelial growth factor ( VEGF) mRNA as well as the changes of related protein expression . Results ①QPCR test showed that eNOS , VEGF mRNA expressions increased in CSWT group compared with control group (4.61 ±0.19 vs 3.99 ±0.17, P<0.05), while compared with CSWT group, eNOS, VEGF mRNA expressions in CSWT +T group were decreased (0.62 ±0.10 vs 0.40 ±0.02, P<0.05), eNOS, VEGF mRNA expressions in CSWT +F group were also decreased (0.53 ±0.02 vs 0.64 ±0.02, P<0.05), all the differ-ences were of statistical significance .②Western blot showed that eNOS , VEGF protein expressions increased in CSWT group compared with control group(0.63 ±0.02 vs 0.43 ±0.02, P<0.05), while compared with CSWT group , eNOS, VEGF protein expressions in CSWT+T group were decreased (0.36 ±0.01 vs 0.29 ±0.02, P<0.05), eNOS, VEGF protein expressions in CSWT +F group were also decreased (0.37 ±0.02 vs 0.30 ±0.02, P<0.05), all the differences were of statistical significance . Conclusion CSWT can improve eNOS , VEGF mRNA and protein expressions in HCMEC cells and FAK and KCa may participate in the signal conduction pathway of CSWT arteriogenesis .

Objective To investigate the results and complications in the treatment of the mechanically unstable fractures of the distal femur when Less Invasive Stabilization System (LISS) is used.Methods From September 2011 to July 2014,81 patients with mechanically unstable fractures of the distal femur were treated with the LISS,according to the inclusion criteria and exclusion criteria,59 patients meet the condition including 31 male and 28 female.The mean age of the patients was 49.8 years (range 18-80 years).The fractures occurred on the left side in 40 cases and on the right side in 19 cases.According to AO classification,27 type 33A2,14 type 33A3,13 type 33C2,5 type 33C3.2 cases combined with ipsilateral fractures of the femoral shaft,according to AO classification,1 type 32A1,1 type 32C1.The mechanism of injury was a fall from the height in 8 cases,a traffic accident in 18 cases,a crush injury in 7 cases,a fall injury in 26 cases.55 fractures were closed,and 4 were open.According to the Gustilo-Anderson classification,there were 3 type Ⅰ,1 type Ⅱ.Results The 7-hole plate were used in 21 patients,9-hole plate in 26 patients and 13-hole plate in 12 cases.The 3.5 mm or 6.5 mm lag screw were placed around the LISS plate to stabilized the articular fracture fragment in 11 cases.The mean operation time was 105.2 min (85-145 min),the mean bleeding volume was 203 ml (130-315 ml).All patients were followed up 11-27 months (average 13.2 months).1 delayed union(13 months),the average time to union was 16 weeks (range 12-21 weeks).The average flexion of the knee was 116 degree,0 degree in extension.The functional outcome:32 had an excellent result,19 had a good result and 8 had a fair result,with 86.4％ excellent and good results.No deep infections occurred.No loss of reduction.3 cases had malalignment,2 failed fixation,4 patients with symptomatic hardware irritation.27 cases underwent implants removal after union,cold-welding occurred in 4 cases (9 screws) which resulted in difficult removal.Conclusion LISS is one of the reliable and effective methods in fixation of mechanically unstable fractures of the distal femur.However,its operation indications and operating instructions should be strictly followed.