Objective To analyze the risk factors and survival status of Epstein-Barr virus(EBV)infection in pa-tients with aplastic anemia(AA)after haploid allogeneic hematopoietic stem cell transplantation(Haplo-HSCT).Methods Clinical data of 78 AA patients who underwent Haplo-HSCT in the hematology department of a hospital from January 1,2019 to October 31,2022 were analyzed retrospectively.The occurrence and onset time of EBV viremia,EBV-related diseases(EBV diseases),and post-transplant lymphoproliferative disorders(PTLD)were ob-served,risk factors and survival status were analyzed.Results Among the 78 patients,38 were males and 40 were females,with a median age of 33(9-56)years old;53 patients experienced EBV reactivation,with a total inci-dence of 67.9％,and the median time for EBV reactivation was 33(13,416)days after transplantation.Among pa-tients with EBV reactivation,49 cases(62.8％)were simple EBV viremia,2 cases(2.6％)were possible EBV di-seases,and 2 cases(2.6％)were already confirmed EBV diseases(PTLD).Univariate analysis showed that age 1＜40 years old at the time of transplantation,umbilical cord blood infusion,occurrence of acute graft-versus-host disease(aGVHD)after transplantation,and concurrent cytomegalovirus(CMV)infection were independent risk fac-tors for EBV reactivation in AA patients after Haplo-HSCT.Multivariate analysis showed that concurrent CMV in-fection was an independent risk factor for EBV reactivation in A A patients after Haplo-HSCT(P=0.048).Ritu-ximab intervention before stem cell reinfusion was a factor affecting the duration of EBV reactivation(P＜0.05).The mortality of EBV viremia,EBV diseases,and PTLD alone were 8.2％,50.0％,and 100％,respectively.The 2-year overall survival rate of patients with and without EBV reactivation were 85.3％,and 90.7％,respectively,difference was not statistically significant(P=0.897).However,patients treated with rituximab had 2-year lower survival rate than those who did not use it,with a statistically significant difference(P=0.046).Conclusion EBV reactivation is one of the serious complications in AA patients after Haplo-HSCT,which affects the prognosis and survival of patients.

Lithocarpus litseifolius is rich in the chalcones phloridzin and trilobatin, the biosynthesis pathways of which have not been fully demonstrated. Chalcone synthase(CHS) is the first key rate-limiting enzyme in the biosynthesis of flavonoids in plants. To explore the functions of CHS gene family in chalcone synthesis of L. litseifolius, this study screened out two CHS genes(LlCHS1 and LlCHS2) from the transcriptome data of this plant, and then bioinformatics analysis and functional characterization were performed for the two genes. The bioinformatics analysis showed that LlCHS1 and LlCHS2 were acidic hydrophilic stable proteins with no transmembrane domain, composed of 395 and 390 amino acid residues, respectively. Both of them contained the characteristic amino acid sequence "WGVLFGFGPGL" and highly conserved active sites(Cys-164, Phe-215, His-303, and Asn-336) of the CHS family. The phylogenetic tree showed that LlCHS1 shared the same clade with similar genes in Aquilaria sinensis, and LlCHS2 was closely related to similar genes in Malus domestica. Under exogenous addition of phloretic acid, co-expression of LlCHS1 or LlCHS2 with Aa4CL from Aromatoleum aromaticum in Escherichia coli catalyzed the production of phloretin from phloretic acid. This study laid a theoretical foundation for revealing the functions of CHS in plants and provided new enzymatic modules for producing phloretin by synthetic biology.
Acyltransferases/chemistry* ; Phylogeny ; Plant Proteins/chemistry* ; Cloning, Molecular ; Amino Acid Sequence

Objective:To evaluate the effects of 3D printing and mixed reality (MR) in teaching ultrasound-guided spinal puncture for scoliosis and to provide new approaches for enhancing educational outcomes in this field.Methods:Twenty anesthesiologists with no experience of spine ultrasound were selected from the Department of Anesthesiology at Peking Union Medical College Hospital and received training in ultrasound-guided spinal puncture in March 2024. They were assigned to the MR glasses+3D printing group or 3D printing group, with 10 participants in each group. The trainees in the MR glasses+3D printing group observed the spine model with the HoloLens 2 glasses and then conducted ultrasound-guided spinal puncture on the 3D-printed spine model. The trainees in the 3D printing group directly conducted ultrasound-guided spinal puncture on the 3D-printed spine model. The teaching effects were evaluated with objective puncture indicators (success rate and puncture time) and questionnaire survey. Data were analyzed using the independent samples t-test or chi-square test in SPSS 27.0. Results:The puncture time was shorter in the MR glasses+3D printing group compared with the 3D printing group [(165.90±69.91) s vs. (239.30±80.30) s, t=2.18, P=0.043]. The success rate of spinal puncture was 90.00% (9/10) in the MR glasses+3D printing group and 80.00% (8/10) in the 3D printing group, with no statistically significant difference ( P>0.999). The satisfaction [(4.70±0.48) vs. (4.80±0.42), t=0.49, P=0.628] and recommendation [(4.80±0.42) vs. (4.90±0.32), t=0.60, P=0.556] of the training method showed insignificant differences between the MR glasses+3D printing group and the 3D printing group. Conclusions:The combination of 3D printing with MR technology can provide trainees with a safe and repeatable high-fidelity simulation training system for ultrasound-guided spinal puncture. The system can improve the confidence and interest of trainees in ultrasound-guided spinal puncture techniques and enhance training efficiency. This combination demonstrates advantages and potential in training for spinal ultrasound and ultrasound-guided spinal puncture.

Objective:To evaluate the effects of 3D printing and mixed reality (MR) in teaching ultrasound-guided spinal puncture for scoliosis and to provide new approaches for enhancing educational outcomes in this field.Methods:Twenty anesthesiologists with no experience of spine ultrasound were selected from the Department of Anesthesiology at Peking Union Medical College Hospital and received training in ultrasound-guided spinal puncture in March 2024. They were assigned to the MR glasses+3D printing group or 3D printing group, with 10 participants in each group. The trainees in the MR glasses+3D printing group observed the spine model with the HoloLens 2 glasses and then conducted ultrasound-guided spinal puncture on the 3D-printed spine model. The trainees in the 3D printing group directly conducted ultrasound-guided spinal puncture on the 3D-printed spine model. The teaching effects were evaluated with objective puncture indicators (success rate and puncture time) and questionnaire survey. Data were analyzed using the independent samples t-test or chi-square test in SPSS 27.0. Results:The puncture time was shorter in the MR glasses+3D printing group compared with the 3D printing group [(165.90±69.91) s vs. (239.30±80.30) s, t=2.18, P=0.043]. The success rate of spinal puncture was 90.00% (9/10) in the MR glasses+3D printing group and 80.00% (8/10) in the 3D printing group, with no statistically significant difference ( P>0.999). The satisfaction [(4.70±0.48) vs. (4.80±0.42), t=0.49, P=0.628] and recommendation [(4.80±0.42) vs. (4.90±0.32), t=0.60, P=0.556] of the training method showed insignificant differences between the MR glasses+3D printing group and the 3D printing group. Conclusions:The combination of 3D printing with MR technology can provide trainees with a safe and repeatable high-fidelity simulation training system for ultrasound-guided spinal puncture. The system can improve the confidence and interest of trainees in ultrasound-guided spinal puncture techniques and enhance training efficiency. This combination demonstrates advantages and potential in training for spinal ultrasound and ultrasound-guided spinal puncture.

Background: Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group. Methods: The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected. Results: Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ² = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response. Conclusion: Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission. Trial Registration NCT01962155; https://clinicaltrials.gov.

Objective To investigate the effect of high-dose coenzyme Q10 on the central nervous system in mice,and to provide experimental basis for clinical safety evaluation.Methods Mice were randomly divided into vehicle control group,perillartine control group,positive control group (chlorpromazine or diazepam) and coenzyme Q10 low,medium and high dose groups (1.5,3.0 and 6.0 g/kg,equivalent to 75,150,and 300 times of clinical dosage,respectively).The corresponding drugs were ig given to mice with the volume of 40 mL/kg.The general behavior of mice was observed directly,the motor coordination ability was observed by rotating stick method,and Anymaze animal behavior video analysis system was used to observe the spontaneous activity of mice and synergistic reaction with sub-threshold dose of pentobarbital sodium.Results There were no significant differences in the general behavioral activity,and the number of spontaneous activity times,mean resident time,and ratio of sleeping were found in all coenzyme Q10 groups,compared with the vehicle and perillartine control groups.Conclusion High dose of coenzyme Q10 has no significantly toxic effect on the central nervous system in mice,which could provide a reliable experimental basis for further medication study and clinical application of high-dose coenzyme Q 10.

Objective To explore the effects of gravitational traction and target regulation of caspase3 on the degenerative intervertebral disc cells in rabbits.Methods Rabbits nucleus pulposus cells transfected with caspase3 siRNA or negative control siRNA were incubated in serum-starved medium for 6 hours and 48 hours.The expression of caspase3 siRNA and cell apoptosis were analyzed by RT-qPCR and Annexin V-fluorescein staining.In order to create intervertebral disc degeneration model,the right anterior side of the annulus fibrosus of lumbar vertebrae of 35 rabbits were damaged by 16-gauge needle.After confirming the success of modeling,35 animal models were randomized into 3 groups:model group (n=10),negative siRNA group (n=10) and caspase3 siRNA group (n=5).Either negative control siRNA or caspase3 siRNA was injected into the center of nucleus pulposus using a 26-gauge needle from the left anterior side,while the model group received no injection.Nucleus pulposus tissue of 5 rabbits selected randomly from every group after 48 hours were analyzed by PCR and Western blotting for caspase3 mRNA and protein expressions.Half of the casepase3 siRNA group were selected randomly and received a routine gravitational traction using a model of our own design,30min per day for 2 weeks,while other groups received no treatment.TNF-o and IL-1β expression levels and histopathological observations were performed after intervention.Results Expression of caspase3 mRNA in rabbit nucleus pulposus cells transfected with caspase3 siRNA decreased obviously in serum-starved medium,and the apoptosis rate of cells cultured in serum-starved medium decreased significantly (P＜0.05).Caspase3 mRNA and caspase3 protein expression of nucleus pulposus injected by caspase3 siRNA down-regulated in the caspase3 siRNA group.Compared with model group and negative control siRNA group,TNF-α and IL-1β expression levels of caspase3 siRNA traction group decreased significantly (P＜0.05),but there was no statistical significance compared with caspase3 siRNA group (P＞0.05).Pathological observation revealed that viable cell number and extracellular matrix contents increased and collagenous fibers arranged regularly in caspase3 siRNA traction group.Conclusion Gravitational traction and target regulation of caspase3 can prevent apoptotic cell death and delay early intervertebral disc degeneration.

OBJECTIVETo develop a dressing with desired antibacterial activity, good water maintaining ability and mechanical properties for wound healing and skin regeneration.

METHODSThe chitosan with different concentrations were added in keratin solution to form porous keratin/chitosan (KCS) scaffolds. The morphological characteristics, chemical composition, wettability, porosity, swelling ratio and degradation of the scaffolds were evaluated. The antibacterial activity was tested by using S. aureus and E. coli suspension for 2 h. And L929 fibroblast cells culture was used to evaluate the cytotoxicity of the KCS scaffolds.

RESULTSThe adding of chitosan could increase the hydrophobicity, decrease porosity, swelling ratio and degradation rate of the KCS porous scaffolds. Mechanical properties of KCS scaffolds could be enhanced and well adjusted by chitosan. KCS scaffolds could obviously decrease bacteria number. The proliferation of fibroblast cells in porous KCS patch increased firstly and then decreased with the increase of chitosan concentration. It was appropriate to add 400 μg/mL chitosan to form porous KCS scaffold for achieving best cell attachment and proliferation compared with other samples.

CONCLUSIONThe porous KCS scaffold may be used as implanted scaffold materials for promoting wound healing and skin regeneration.

Anti-Bacterial Agents ; administration & dosage ; Bandages ; Cell Line ; Cell Proliferation ; Chitosan ; Fibroblasts ; cytology ; Humans ; Keratins ; Microscopy, Electron, Scanning ; Porosity ; Spectroscopy, Fourier Transform Infrared ; Wound Healing

Bel1, a transactivator of prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have shown that Bel1 bears a nuclear localization signal (NLS), but its amino acid sequence remains unclear and the corresponding importins have not been identified. In this report, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment could direct nuclear localization, which accords with the consensus sequence K(K/R)X(K/R) of monopartite NLS. Point mutation experiments revealed that K218, R219, and R221 are essential for the nuclear localization of Bel1. The results of the GST-pulldown showed that the Bel1 fragment with residues 215-223, which bears the NLS, interacts with KPNA1, KPNA6, and KPNA7. This result suggests that KPNA1, KPNA6, and KPNA7 maybe involved in Bel1 nuclear translocation.
Cell Line ; Cell Nucleus ; genetics ; metabolism ; virology ; Humans ; Nuclear Localization Signals ; genetics ; metabolism ; Protein Binding ; Protein Transport ; Retroviridae Infections ; genetics ; metabolism ; virology ; Retroviridae Proteins ; chemistry ; genetics ; metabolism ; Spumavirus ; chemistry ; genetics ; physiology ; Trans-Activators ; chemistry ; genetics ; metabolism ; alpha Karyopherins ; genetics ; metabolism