Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

Glucose oxidase (GOD) is an oxygen-consuming dehydrogenase that can catalyze the production of gluconic acid hydrogen peroxide from glucose, and its specific mechanism of action makes it promising for applications, while the low catalytic activity and poor thermostability have become the main factors limiting the industrial application of this enzyme. In this study, we used the glucose oxidase AtGOD reported with the best thermostability as the source sequence for phylogenetic analysis to obtain the GOD with excellent performance. Six genes were screened and successfully synthesized for functional validation. Among them, the glucose oxidase AhGODB derived from Aspergillus heteromorphus was expressed in Pichia pastoris and showed better thermostability and catalytic activity, with an optimal temperature of 40 ℃, a specific activity of 112.2 U/mg, and a relative activity of 47% after 5 min of treatment at 70 ℃. To improve its activity and thermal stability, we constructed several mutants by directed evolution combined with rational design. Compared with the original enzyme, the mutant T72R/A153P showcased the optimum temperature increasing from 40 to 50 ℃, the specific activity increasing from 112.2 U/mg to 166.1 U/mg, and the relative activity after treatment at 70 ℃ for 30 min increasing from 0% to 33%. In conclusion, the glucose oxidase mutants obtained in this study have improved catalytic activity and thermostability, and have potential for application.
Glucose Oxidase/chemistry* ; Enzyme Stability ; Aspergillus/genetics* ; Pichia/metabolism* ; Temperature ; Catalysis ; Fungal Proteins/metabolism* ; Hot Temperature

Mercury (Hg)-contaminated soil poses a significant threat to the environment and human health. Hg-resistant microorganisms have the ability to survive under the stress of inorganic and organic Hg and effectively reduce Hg levels and toxicity. Compared to physical and chemical remediation methods, microbial remediation technologies have garnered increasing attention in recent years due to their lower cost, remarkable efficacy, and minimal environmental impact. This paper systematically elucidates the molecular mechanisms of Hg resistance in microbes, with a focus on their potential applications in phytoremediation of Hg-contaminated soils through plant-microbe interactions. Furthermore, it highlights the critical role of microbes in enhancing the effectiveness of transgenic plants for Hg remediation, aiming to provide a theoretical foundation and scientific basis for the bioremediation of Hg-contaminated soils.
Mercury/toxicity* ; Biodegradation, Environmental ; Soil Pollutants/isolation & purification* ; Soil Microbiology ; Plants, Genetically Modified/metabolism* ; Bacteria/genetics*

Objective:To evaluate the role of minimal residual disease (MRD) monitoring during early induction therapy for the treatment of childhood acute lymphoblastic leukemia (ALL).Methods:This was a multicenter retrospective cohort study. Clinical data of 1 164 ALL patients first diagnosed between October 2016 and June 2019 was collected from 16 hospitals in South China Children′s Leukemia Group. According to MRD assay on day 15 of early induction therapy, they were divided into MRD<0.10% group, MRD 0.10%-<10.00% group and MRD≥10.00% group. According to MRD assay on day 33, they were divided into MRD<0.01% group, MRD 0.01%-<1.00% group and MRD≥1.00% group. Age, onset white blood cell count, central nervous system leukemia (CNSL), molecular genetic characteristics and other data were compared between groups. Kaplan-Meier method was used for survival analysis. Cox regression model was used to analyze prognostic factors.Results:Of the 1 164 enrolled patients, there were 692 males and 472 females. The age of diagnosis was 4.7 (0.5, 17.4) years. The white blood cell count at initial diagnosis was 10.7 (0.4, 1 409.0) ×10 9/L. Among all patients, 53 cases (4.6%) had CNSL. The follow-up time was 47.6 (0.5, 68.8) months. The 5-year overall survival (OS) and 5-year relapse-free survival (RFS) rates were (93.1±0.8) % and (90.3±1.1) %. On day 15 of early induction therapy, there were 466 cases in the MRD<0.10% group, 523 cases in the MRD 0.10%-<10.00% group and 175 cases in the MRD≥10.00% group. The 5-year OS rates of the MRD<0.10% group, MRD 0.10%-<10.00% group and MRD≥10.00% group were (95.4±1.0) %, (93.3±1.1) %, (85.4±2.9) %, respectively, while the RFS rates were (93.2±1.6) %, (90.8±1.4) %, (78.9±4.3) %, respectively ( χ2=16.47, 21.06, both P<0.05). On day 33 of early induction therapy, there were 925 cases in the MRD <0.01% group, 164 cases in the MRD 0.01%-<1.00% group and 59 cases in the MRD≥1.00% group. The 5-year RFS rates in the MRD 0.01%-<1.00% group was lowest among three groups ((91.4±1.2) % vs. (84.5±3.2) % vs. (87.9±5.1) %). The difference between three groups is statistically significant ( χ2=9.11, P=0.010). Among ALL patients with MRD≥10.00% on day 15 of induction therapy, there were 80 cases in the MRD <0.01% group on day 33, 45 cases in the MRD 0.01%-<1.00% group on day 33 and 45 cases in the MRD≥1.00% group on day 33. The 5-year RFS rates of three groups were (83.9±6.0)%, (67.1±8.2)%, (83.3±6.9)% respectively ( χ2=6.90, P=0.032). Univariate analysis was performed in the MRD≥10.00% group on day 15 and the MRD 0.01%-<1.00% group on day 33.The 5-year RFS rate of children with CNSL was significantly lower than that without CNSL in the MRD≥10.00% group on day 15 ((50.0±20.4)% vs. (80.3±4.4)%, χ2=4.13, P=0.042). Patients with CNSL or MLL gene rearrangement in the MRD 0.01%-<1.00% group on day 33 had significant lower 5-year RFS rate compared to those without CNSL or MLL gene rearrangement ((50.0±25.0)% vs. (85.5±3.1)%, χ2=4.06, P=0.044;(58.3±18.6)% vs. (85.7±3.2)%, χ2=9.44, P=0.002). Multivariate analysis showed that age ( OR=0.58, 95% CI 0.35-0.97) and white blood cell count at first diagnosis ( OR=0.43, 95% CI 0.27-0.70) were independent risk factors for OS. The MRD level on day 15 ( OR=0.55,95% CI 0.31-0.97), ETV6-RUNX1 fusion gene ( OR=0.13,95% CI 0.03-0.54), MLL gene rearrangement ( OR=2.55,95% CI 1.18-5.53) and white blood cell count at initial diagnosis ( OR=0.52,95% CI 0.33-0.81) were independent prognostic factors for RFS. Conclusions:The higher the level of MRD in early induction therapy, the worse the OS. The MRD levels on day 15 is an independent prognostic factor for RFS.The MRD in early induction therapy guided accurate risk stratification and individualized treatment can improve the survival rate of pediatric ALL.

Objective To investigate the effectiveness of team-based learning(TBL)combined with research-based learning(RBL)in enhancing evidence-based nursing practice skills of undergraduate nursing interns.Methods A total of 114 undergraduate nursing students who interned in the operating room of a ⅢA hospital in Guangzhou from July 2021 to April 2022 were selected as study subjects.A randomized cluster sampling method based on a random number table was used to divide the students into a control group and a trial group.The control group received traditional teaching methods,while the trial group was taught using a combination of TBL and RBL.The two groups were compared in terms of evidence-based practice skills,critical thinking abilities,and their evaluations on the teaching methods.Results The differences in evidence-based practice skills and critical thinking abilities before and after the internship were significantly higher in the observation group compared to the control group(t=35.108,35.897;both P<0.05).Additionally,post-internship evaluation scores for the teaching methods in the trial group were significantly higher than those in the control group(t=-17.580,P<0.05).Conclusion TBL combined with RBL effectively enhances the evidence-based nursing practice skills and critical thinking abilities of undergraduate nursing interns.This approach also improves students'evaluations on the teaching methods and fosters the cultivation of excellent clinical evidence-based nursing professionals.

Human parvovirus B19 (HPVB19) belongs to Parvoviridae, a genus of erythrovirus, and has been associated with various human diseases, and HPVB19 infection is one of the most important causes of refractory anemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). This study retrospectively analyzed 24 patients with HSCT combined with HPVB19 infection to collate and summarize the clinical presentation, treatment, and regression of patients with combined HPVB19 infection after allo-HSCT and provide experience in the management of HPVB19 infection after allo-HSCT. The median age of the patients with HPVB19 infection was 25 years, and the median time of infection occurrence was +107 days after transplantation, and 22 (91.7% ) had anemia with a median hemoglobin (HGB) level of 77.5 (46-149) g/L, and 13 (54.2% ) had new-onset anemia or persistent decline in HGB. The median length of hospital stay was 19 days. Among patients with new-onset anemia or persistent decline in HGB, the mean increase in HGB after treatment with intravenous immunoglobulin and/or antiviral therapy was 15.69 g/L, and treatment was effective in 10 (76.92% ) patients. HPVB19 infection should be alerted to the development of refractory anemia after HSCT; despite the lack of specific treatment, the overall prognosis of HPVB19-infected patients is good.

Hepatosplenic candidiasis (HSC) is a rare type of candidiasis that can occur in patients with hematologic malignancies, hematopoietic stem cell transplantation. At present, there is still a lack of studies on HSC in patients with hematologic disorders. Based on The Chinese Guidelines for the Diagnosis and Treatment of Invasive Fungal Disease in Patients with Hematological Disorders and Cancers (the 6th revision), We retrospectively analyzed the clinical characteristics and prognosis of patients with HSC treated in Peking University Institute of Hematology from 2008 to 2022. Finally, eighteen patients were included, with 1 (5.6%) proven, 2 (11.1%) probable, and 15 (83.3%) possible HSC. Among them, 3 (16.7%) patients occurred after haploid hematopoietic stem cell transplantation and 15 (83.3%) patients occurred after chemotherapy. 6 (33.3%) patients had positive blood cultures, including 4 cases of Candida tropicalis and 2 cases of Candida albicans. At 4 weeks of antifungal therapy, 10 (58.8%) patients achieved partial response (PR), At 8 weeks, 1 (6.3%) patients achieved complete response and 10 (62.5%) patients achieved PR. At 6 months after diagnosis, 3 (16.7%) patients died of hematopoietic recurrence, and none of them died of HSC. As a rare fungal infection disease, HSC has a low positive rate of microbiological and histological examinations, a persistent treat cycle, and has difficulty in remission, reminding us of the need for vigilance in patients with hematopoietic disorders and persistent fever.

OBJECTIVE To investigate the current situation of the review and comment on inpatient medical orders in Beijing municipal hospitals， and to put forward countermeasures and suggestions for further improving related work. METHODS A questionnaire survey was conducted to investigate the current situation of the review and comment on inpatient medical orders in 22 Beijing municipal hospitals. The statistical analysis was conducted for the survey results. RESULTS A total of 22 questionnaires were distributed， with recovery effective rate of 100%. The 22 hospitals carried out inpatient medical order comment， but their proportion varied among hospitals （0.88%-98.54%）； medical order comment mainly focused on antibiotics， proton pump inhibitors， anesthetic drugs/class Ⅰ psychotropic drugs， auxiliary drugs and other categories； 205 pharmacists participated in the comment of inpatient medical orders， most of whom hold intermediate or higher professional titles （89.27%）； 21 hospitals conducted inpatient medical order comment and feedback the results to relevant departments/responsible persons， but the intervention situation was not the same. Eighteen hospitals had carried out the review of inpatient medical orders； reviewed drug category was roughly the same as the category involved in the medical order comment； review content involved the suitability of administration routes， dosage， etc. The review was conducted mainly through the cooperation of audit software and pharmacists. CONCLUSIONS The comment and review of inpatient medical orders in Beijing municipal hospitals carried out in an orderly manner， and preliminary results have been achieved； at the same time， it is necessary to further increase the ability of participating pharmacists， improve audit standards， optimize pre-audit （No.ZYLX201805） software， and promote rational drug use among hospitalized patients through doctor-pharmacist collaboration.

Objective To investigate the association between abdominal visceral fat content and the occur-rence of type 2 diabetes mellitus(T2DM)and the potential of predicting the incidence of T2DM with abdominal visceral fat content.Methods The study included 45 patients with T2DM who underwent 3.0T MRI and 80 control patients,recruited from the First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology.IDEAL software was utilized for automated image recombination to obtain fat fraction,relaxation rate(R2*),lipid phase,water phase,and in-phase images.Fat fraction images were transferred to AW 4.6 workstation for measurement of fat fraction plots by two observers using a double-blind approach.ITK-SNAP software was employed for semi-automatic delineation of abdominal visceral fat and subcutaneous adipose tissue through threshold segmentation method,followed by volume calculation.Statistical analysis of the data was performed using SPSS 26.0 software.Results(1)The T2DM group exhibited significantly higher levels of abdominal visceral adipocyte volume(VATV),hepatic fat fraction(HFF),pancreatic fat fraction(PFF),triglyceride(TG),and fasting blood glucose(FBG)compared to the control group.Conversely,the high density lipoprotein(HDL)level was significantly lower in the T2DM group than in the control group(P＜0.05).The correlation analysis revealed that in male patients,abdominal VATV exhibited positive associations with body mass index(BMI),waist circumference,subcutaneous adipose tissue volume(SATV),hepatic fat fraction(HFF),TG,and FBG;while displaying a negative association with HDL.In female patients,VATV demonstrated positive correlations with BMI,waist circumference,SATV,and HFF.(2)The ROC curve analysis demonstrated that the area under the curve(AUC)for PFF in diagnosing T2DM was 0.656(P＜0.01,95%CI:0.534～0.777).The determined cut-off value was 8.44%,yielding a sensitivity of 50%and specificity of 82.5%.Similarly,HFF exhibited an AUC of 0.744(P＜0.05,95%CI:0.637～0.851),with a cut-off value of 2.99%,sensitivity of 97.5%,and specificity of 42%.In male patients,VATV displayed an optimal threshold for T2DM diagnosis at a volume of approximately 3 466 cm3,resulting in a sensitivity of 63.2%,specificity of 89.5%,and AUC of 0.78(P＜0.01).The sensitivity was 78.9%and specificity was 84.2%when SATV,PFF and HFF were combined and the AUC was 0.839(P＜0.01).The optimal threshold for diagnosing T2DM with VATV in female patients was 2 103cm3,and the sensitivity,specificity and AUC of T2DM with VATV were 66.7%,66.7%and 0.68(P＜0.05).When SATV,PFF and HFF were combined,the sensitivity was 90.5%.The specificity was 76.2%and the AUC was 0.909(P＜0.01).(3)For each additional unit of TG and FBG,the risk of T2DM respectively increased by 3986.3%and 417.1%,in other words,TG and FBG were independent predictors of the occurrence of T2DM.Conclusion The predictive value of pancreatic fat fraction,liver fat fraction,and abdominal visceral fat tissue volume for the onset of type 2 diabetes is limited.However,combining abdominal visceral adipose tissue volume with subcutaneous adipose tissue volume,pancreatic fat fraction,and liver fat fraction significantly enhances the efficacy,sensitivity,and specificity of predicting type 2 diabetes occurrence.The IDEAL-IQ technique offers a convenient method for quantitatively evaluat-ing intra-abdominal ectopic adipose tissue deposition in patients with type 2 diabetes mellitus.It demonstrates advantages such as simplicity,good stability,high reproducibility and holds great potential.