The α-amino acid ester acyltransferase (SAET) from Sphingobacterium siyangensis is one of the enzymes with the highest catalytic ability for the biosynthesis of l-alanyl-l-glutamine (Ala-Gln) with unprotected l-alanine methylester and l-glutamine. To improve the catalytic performance of SAET, a one-step method was used to rapidly prepare the immobilized cells (SAET@ZIF-8) in the aqueous system. The engineered Escherichia coli (E. coli) expressing SAET was encapsulated into the imidazole framework structure of metal organic zeolite (ZIF-8). Subsequently, the obtained SAET@ZIF-8 was characterized, and the catalytic activity, reusability and storage stability were also investigated. Results showed that the morphology of the prepared SAET@ZIF-8 nanoparticles was basically the same as that of the standard ZIF-8 materials reported in literature, and the introduction of cells did not significantly change the morphology of ZIF-8. After repeated use for 7 times, SAET@ZIF-8 could still retain 67% of the initial catalytic activity. Maintained at room temperature for 4 days, 50% of the original catalytic activity of SAET@ZIF-8 could be retained, indicating that SAET@ZIF-8 has good stability for reuse and storage. When used in the biosynthesis of Ala-Gln, the final concentration of Ala-Gln reached 62.83 mmol/L (13.65 g/L) after 30 min, the yield reached 0.455 g/(L·min), and the conversion rate relative to glutamine was 62.83%. All these results suggested that the preparation of SAET@ZIF-8 is an efficient strategy for the biosynthesis of Ala-Gln.
Escherichia coli/genetics* ; Glutamine ; Zeolites/chemistry* ; Amino Acids

Objective:To investigate the relationship between NUDT15 gene polymorphism and tolerance to treatment with 6-mercaptopurine (6-MP) in children with acute lymphoblastic leukemia (ALL).Methods:Fifty-eight children diagnosed with ALL in Shanxi Children's Hospital from January 2019 to December 2020 were recruited. All of them were treated with CCLG-ALL2018 chemotherapy regimen and the bone marrow showed complete remission. They received 6-MP oral treatment during maintenance treatment. Single nucleotide polymorphism of NUDT15 gene was detected by real-time fluorescence quantitative polymerase chain reaction. The bone marrow suppression after 6-MP treatment and 6-MP tolerance dose in patients with different NUDT15 genotypes were analyzed.Results:Among 58 patients, 3 patients had NUDT15 TT genotype, 46 patients had CC genotype and 9 patients had TC genotype. During maintenance treatment with 6-MP, the differences in leukocyte count, hemoglobin and platelet count among the three groups of patients with different NUDT15 genotypes were statistically significant (all P < 0.05). Among 58 patients, 23 (39.66%) patients had varying degrees of neutropenia after medication, including 16 cases of NUDT15 CC genotype, 5 cases of TC genotype and 2 cases of TT genotype. There was a statistically significant difference in bone marrow suppression among the three groups ( H = 29.10, P < 0.05). The dosages of 6-MP used in patients with TT, CC and TC genotypes were (10.4±8.8) mg·m -2·d -1, (41.5±1.3) mg·m -2·d -1 and (36.7±2.4) mg·m -2·d -1, respectively, and the difference was statistically significant ( F = 16.95, P < 0.05). Conclusions:Children with different NUDT15 genotypes have different tolerance to 6-MP, and NUDT15 gene polymorphism is associated with 6-MP intolerance during maintenance treatment in children with ALL, which may affect the treatment of the disease.

Objective:To evaluate the short-term efficacy and the improvement of quality of life of enzyme replacement therapy (ERT) with Imiglucerase on children with Gaucher disease(GD) through the same time monitoring.Methods:Six children diagnosed as GD who were treated by ERT with Imiglucerase in the Department of Hematology of the Children′s Hospital of Shanxi Province from May 2019 to May 2020 were recruited.Every 3 months, the sizes of the liver and spleen was palpated, the change of bone pain was recorded, and the haematological index was examed.The volumes of the liver and spleen at 1-year treatment were measured by CT.Bone involvement was examined by magnetic resonance imaging (MRI). In addition, the body weight, height, and the 36-Item Short Form Survey (SF-36) were measured and compared with pre-treatment levels.These data were analyzed statistically by SPSS 25.0 and the difference between pretherapy and post-treatment was compared by paired t test. Results:Six children diagnosed as GD received ERT with Imiglucerase.No adverse events were reported.Decreased volumes of the liver and spleen, and increased hemoglobin level and platelet count were detected after 3-6 months of ERT.After 1 year of ERT, hemoglobin level significantly increased compared with pre-treatment level ( t=4.200, P=0.008). Although the platelet count increased at 1-year ERT, it was comparable with pre-treatment level ( t=2.260, P=0.073). The volumes of liver and spleen decreased by (22.10±15.28)% ( t=2.725, P=0.042) and (47.10±18.42)% ( t=3.162, P=0.034) after 1 year of ERT, respectively.During the first year of ERT, the height and weight increased (6.17±2.86) cm ( t=5.286, P=0.003) and (4.08±2.01) kg ( t=4.975, P=0.004), respectively.SF-36 score increased significantly from (489.35±103.99) points to (632.75±73.34) points ( t=5.740, P=0.002). After 1 year of ERT, 1 patient still had bone pain, and 2 cases were worse in bone MRI, which may be attributed to the short period of follow-up and insufficient dose, and another 3 had no change in bone MRI. Conclusions:ERT ameliorates GD-associated anemia, organomegaly and growth retardation, and improves the growth rate of body mass and height and the quality of life in the short period.However, short-term ERT does not improve the bone disease.

Objective:To investigate the reasonable dosage of heparin anticoagulation scheme during plasma adsorption (PA) therapy for liver failure.Methods:Patients with liver failure treated with PA therapy were retrospectively collected and divided according to the anticoagulation scheme into the first-dose heparin anticoagulation group and the first-dose plus maintenance heparin anticoagulation group. Clinical data and laboratory test results were compared before and after treatment between the two groups. Paired t-tests were used for comparison within the normally distributed groups. An independent two-sample t-test was used for inter group comparison. Wilcoxon rank-sum test was used for measurement data that did not conform to a normal distribution. Fisher's exact test was used to compare the count data between groups. Results:There were 138 cases with liver failure treated with PA therapy from October 2017 to September 2020. Among them, 83 and 55 cases were in the first-dose heparin anticoagulation and first-dose plus maintenance heparin anticoagulation group, respectively. Age, gender, and laboratory data before treatment were comparable between the two groups. PA treatment was successfully completed in both groups of patient, and there was no statistically significant difference in the determination of coagulation level with plasma separators ( Z=-0.15, P=0.216). There were different degrees of bleeding complications in both groups. In the first-dose heparin anticoagulation group, there were two cases (2.4%) of central venous catheter bleeding and one case (1.2%) of epistaxis. In the first-dose plus maintenance heparin anticoagulation group, there were five cases (9.1%) of central venous catheter bleeding, two cases (3.6%) of skin bleeding, one case (1.8%) of epistaxis, and one case (1.8%) of upper gastrointestinal bleeding. The incidence of bleeding complications was lower in the first-dose of heparin anticoagulation than first-dose plus maintenance heparin anticoagulation group, and the difference was statistically significant ( P<0.001). The activated partial thromboplastin time of the two groups was prolonged after therapy withdrawal than with therapy, and the difference was statistically significant (first-dose heparin anticoagulation group: t=3.850, P=0.022; first-dose plus maintenance heparin anticoagulation group: t=6.733, P=0.007). The activated partial thromboplastin time was prolonged in patients with first-dose plus maintenance heparin anticoagulation than first-dose heparin anticoagulation group, and the difference was statistically significant ( P=0.025). The total bilirubin of the two groups before and after PA was significantly changed (the first-dose heparin anticoagulation group: Z=-2.455, P=0.017; the first-dose plus maintenance heparin anticoagulation group: Z=-2.307, P=0.024), and there was no statistically significant difference between the two groups ( P=0.412). There was no statistically significant difference in platelet changes before and after PA therapy between the two groups (the first dose of heparin anticoagulation group: Z=-0.529, P=0.480; the first-dose plus maintenance heparin anticoagulation group: Z=-0.276, P=0.362). Conclusion:Anticoagulation scheme without maintenance medication is feasible with prothrombin activity before ≤20-40%, activated partial thromboplastin time of ≤87 s (2 times the upper normal value), platelet count before treatment (excluding contraindications to heparin) ≥50×10 9/L, and the first dose of heparin administration of 0.2 mg/kg during PA therapy in patients with liver failure.

Objective:To construct a risk nomogram early prediction model for myasthenia gravis (MG) after thymoma resection and to verify the predictive performance of the model.Methods:Using the convenient sampling method, a total of 477 patients undergoing thymoma resection who were admitted to Department of Thoracic Surgery in Beijing Tongren Hospital Affiliated to Capital Medical University from March 2018 to February 2021 were selected as the training set. A total of 62 patients who underwent thymoma resection from March 2017 to February 2018 were set as the validation set for retrospective analysis. The clinical data of the included patients were analyzed. Logistic regression analysis was used to explore the independent risk factors of MG after thymoma resection and a risk nomogram prediction model was constructed.Results:The incidence of postoperative MG in 477 patients who underwent thymoma resection was confirmed by chest X-ray, CT and related tests, which was 14.05% (67/477) . There were statistical differences between the two groups of patients with MG and those without MG in terms of combination of immune diseases, preoperative course of disease, surgical route, complete tumor resection, WHO pathological classification, postoperative pulmonary infection, postoperative radiotherapy and chemotherapy and other data ( P<0.05) . Logistic regression analysis showed that combined immune disease, thoracotomy, incomplete tumor resection, WHO pathological classification of A+AB, postoperative pulmonary infection and no postoperative chemoradiotherapy were independent risk factors for MG after thymoma resection ( P<0.05) . Based on 6 independent risk factors, a risk nomogram prediction model of MG after thymoma resection was established. The results showed that the C- index of the training set and the validation set were 0.837 (95% CI: 0.807-0.867) and 0.817 (95% CI: 0.807-0.867) , respectively. The calibration curves for both sets showed good fit to the ideal curve, with areas under the receiver operating characteristic curves of 0.834 (95% CI: 0.794-0.874) and 0.825 (95% CI: 0.789-0.861) , respectively. Conclusions:Combination of immune diseases, surgical approach of thoracotomy, incomplete tumor resection, WHO pathological classification of A+AB, postoperative pulmonary infection and no postoperative radiotherapy and chemotherapy are independent risk factors for MG after thymoma resection. The risk nomogram prediction model established based on the above risk factors can accurately assess and quantify the risk of MG after thymoma resection, which has good predictive ability.

Objective:To explore the clinical characteristics and prognostic factors of children with relapsed acute lymphoblastic leukemia (ALL).Methods:The clinical data of 52 children with relapsed ALL in Children's Hospital of Shanxi Province from January 2010 to April 2019 were retrospectively analyzed. The clinical characteristics of the children were summarized and the prognostic factors after recurrence were analyzed.Results:Till May 1, 2019, 5 out of 52 children gave up treatment after diagnosis and were lost to follow-up. For the remaining 47 children with successful follow-up, the median age at initial diagnosis was 60 months (11-168 months), the median time from initial diagnosis to relapse was 21 months (2-112 months), the median follow-up time was 5.5 months (1.0-69.0 months), and the 2-year overall survival (OS) rate after relapse was 31%. Nine patients accepted allogeneic hematopoietic stem cell transplantation after the second time complete remission, the median time from diagnosis to transplantation was 4.5 months (3.0-7.0 months), and the median follow-up time was 22 months (4-69 months). The 2-year OS rates in relapsed children with white blood cell count < 50×10 9/L and ≥ 50×10 9/L at initial diagnosis were 39% and 13%, respectively (χ 2=5.623, P=0.018). The 2-year OS rate after relapse in standard-risk, intermediate-risk and high-risk groups were 72%, 31% and 8%, respectively (χ 2=10.068, P=0.007). The 2-year OS rate after relapse in very early relapse, early relapse and late relapse groups were 0, 33% and 79%, respectively (χ 2=30.066, P < 0.01). The 2-year OS rate after relapse in chemotherapy with or without radiotherapy group, transplantation group and irregular treatment group were 57%, 89% and 0, respectively (χ 2=26.885, P < 0.01). Cox multivariate analysis showed that relapse time was the independent risk factor affecting the prognosis of children with relapsed ALL ( HR=0.340, 95% CI 0.146-0.789, P=0.012). Compared with the transplantation group, the risk of death in the chemotherapy with or without radiotherapy group and the irregular treatment group was significantly higher ( HR=12.313, 95% CI 1.266-119.758, P=0.031; HR=20.699, 95% CI 2.230-192.129, P=0.008), suggesting that hematopoietic stem cell transplantation is a protective factor for the prognosis of children with relapsed ALL. Conclusions:The relapse of ALL in children mainly happens in very early and early time. The main part of relapse is bone marrow, and there are many high-risk patients at initial diagnosis. The risk group at initial diagnosis, white blood cell count at initial diagnosis, relapse time, and treatment after relapse are the risk factors affecting the prognosis, and the relapse time and hematopoietic stem cell transplantation are the independent prognostic factors.

Objective: To investigate the current status of family hardiness and its influencing factors in children with leukemia. Methods: The children with leukemia and their caregivers in the Children's Hospital of Shanxi from August to November 2017 were enrolled. A questionnaire survey was conducted using a convenient sampling method, and 100 questionnaires were distributed. The questionnaire included the General Status Questionnaire, the Family Hardiness Index (FHI), the Coping Health Inventory for Parents (CHIP) and the Positive and Negative Affect Scale (PANAS). Results: A total of 92 valid questionnaires were collected. Among the scores of family hardiness in children with leukemia [(3.29±0.43) points], the responsibility score [(3.32±0.45) points] was higher than the control score [(3.31±0.46) points] and the challenge score [(3.23±0.53) points], and the challenge score was the lowest. The score of frequency of coping styles used by the caregiver of the child with leukemia was (3.64±0.70) points, the most frequent coping style used by the caregiver was "family unity, optimism, cooperative attitude" [(3.73±0.89) points], and the positive [(3.28±0.84) points] and negative [(2.51±0.80) points] emotions were in a moderate state. The child's sex, age, stage of chemotherapy and medical insurance status, the caregiver of the child, the age of the caregiver, the family's place of residence, and the education level were the related factors affecting the family hardiness score (all P < 0.01). The age of child, CHIP-1, CHIP-2, positive emotion and negative emotion were independent factors affecting the family hardiness (all P < 0.05). The CHIP, CHIP-1, CHIP-2 and positive emotion were positively correlated with the family hardiness (r values were 0.827, 0.883, 0.707 and 0.846, all P < 0.01); the negative emotion was negatively correlated with the family hardiness (r=-0.832, P < 0.01). Conclusion The family hardiness of children with leukemia is in the middle and upper level, the children's age, caregiver's coping style, positive emotion and negative emotion are factors affecting the family hardiness.

Objective: To investigate the molecular characterization of adult diarrhea cases caused by enterotoxic Escherichia coli (ETEC) and explore the practical model of epidemiology for laboratory technique and data needs based on the surveillance network. Methods: Epidemiological design and sampling targeted adult cases ETEC caused diarrhea in epidemic season. The enterotoxin type, serogroup, resistance, colonization factor and molecular type of ETEC were identified. Multiple dynamic phenotypic characteristics of ETEC were indicated by multidimensional and multivariable data. Results: From 2016 to 2018, 84 eligible ETEC strains were detected. The dominant serums/toxins were O∶6 (STh), O∶25 (LT), O∶159 (STh), O∶153 (STh). O∶6 (STh+CS21), which replaced O∶25 and O∶159 as the popular clones in 2018. Six cases of O∶153 (STh+CFA/I+CS8+PT34) in outbreak in 2017 were imported ones. The resistance rates of ETEC strains detected in adults to sulfasoxazole, naproxinic acid, ampicillin and azithromycin were more than 30%, multidrug resistance (MDR) reached 58.3%. Serum/toxin types suggested that attenuated strains were more likely to become MDR. Molecular typing confirmed that the genetic similarity of the dominant clone of O∶6 serogroup (PT20-24) was higher than O∶25 and O∶159. There was a high correlation between the minimal inhibitory concentration (MIC) of azithromycin and the resistant gene mphA (87.5%, 28/32). O∶6 (STh+CS21+mphA) resistant clone was first detected in 2016. Conclusion A new epidemic clone in adult ETEC diarrhea cases in Shanghai was O∶6 (STh+CS21+mphA). For the first time the association between azithromycin resistance gene mphA and a serum group of ETEC was observed. Multidimensional and multivariate analysis techniques based on epidemiology can help reveal the potential transmission pattern of ETEC for the accurate surveillance and early warning of outbreaks.

Objective To explore the nursing methods for the treatment of lung cancer by thoracoscopy.Methods A total of 120 patients with lung cancer in our hospital were divided into control group and study group by random numbers table method,with 60 cases in each group.The control group took routine perioperative nursing intervention,while the control group implemented comprehensive nursing intervention.The operation time,length of hospital stay and postoperative complications of the two groups were observed and compared.Results The hospitalization time of the study group was lower than that of the control group (P ＜ 0.05).There was no significant difference in operation time between the two groups (P ＞ 0.05).The incidence of postoperative complications was 1.67％ in the study group,which was lower than 13.33％ in the control group (P ＜0.05).Conclusion Comprehensive nursing can reduce the incidence of postoperative complications,shorten the length of hospital stay,so it is suitable for clinical application.

Objective To explore the nursing methods for the treatment of lung cancer by thoracoscopy.Methods A total of 120 patients with lung cancer in our hospital were divided into control group and study group by random numbers table method,with 60 cases in each group.The control group took routine perioperative nursing intervention,while the control group implemented comprehensive nursing intervention.The operation time,length of hospital stay and postoperative complications of the two groups were observed and compared.Results The hospitalization time of the study group was lower than that of the control group (P ＜ 0.05).There was no significant difference in operation time between the two groups (P ＞ 0.05).The incidence of postoperative complications was 1.67％ in the study group,which was lower than 13.33％ in the control group (P ＜0.05).Conclusion Comprehensive nursing can reduce the incidence of postoperative complications,shorten the length of hospital stay,so it is suitable for clinical application.