Objective:To evaluate the clinical value of a capillary electrophoresis-based method for gene diagnosis of hyperphenylalaninemia.Methods:In this single-center prospective study, 40 newborns with suspected hyperphenylalaninemia detected by neonatal liquid chromatography-tandem mass spectrometry screening at Nanjing Maternity and Child Health Care Hospital from February 2021 to February 2023 were included, with 22 males, 18 females and a mean age at diagnosis of 21.93 days.Capillary electrophoresis was used to detect 85 variants of the phenylalanine hydroxylase ( PAH) gene in 40 newborns with suspected hyperphenylalaninemia.The PAH gene of undiagnosed patients was further analyzed by Sanger sequencing.The detection rate, sensitivity and specificity of capillary electrophoresis were calculated. Results:Among these 40 newborns with suspected hyperphenylalaninemia, 71 PAH variants were detected by capillary electrophoresis, 32 patients were clearly diagnosed, only 1 pathogenic variant was found in 5 patients, and no pathogenic variant was found in the last 3 patients.Therefore, the detection rate, sensitivity and specificity of capillary electrophoresis for analysis of the PAH gene were 80.00%, 88.75% and 100%, respectively. Conclusions:The capillary electrophoresis-based method can rapidly, efficiently and accurately detect PAH gene variants at lower cost and is a promising gene detection method for hyperphenylalaninemia in clinical practice.

Objective To investigate the diagnostic value of biparametric magnetic resonance imaging(bpMRI)radiomics combined with prostate-specific antigen density(PSAD)in predicting low-grade and high-grade prostate carcinoma(PCa).Methods The clinical and imaging data of patients with PCa confirmed by pathology in Taizhou Central Hospital from June 2018 to October 2022 were retrospectively analyzed.According to Gleason grade group(GGG),GGG≤2 was defined as low-grade PCa,and GGG＞2 was defined as high-grade PCa.PCa patients with different grades were randomly divided into training group and test group according to a ratio of 7∶3.Radiomics features were extracted based on T2 weighted imaging(T2WI)and apparent diffusion coefficient(ADC)sequences.Feature selection and dimensionality reduction were carried out using maximum relevance minimum redundancy,least absolute shrinkage and selection operator,and 5-fold cross validation was performed to retain the best radiomics features.Receiver operating characteristic(ROC)curve and Delong's test were used to evaluate the performance of each model.Decision curve analysis(DCA)was used to evaluate the clinical utility of the model.Results Among all the models,T2WI-ADC-PSAD combined model had the best diagnostic efficiency,the area under the curve(AUC)in training group and test group were 0.882,0.772,respectively.Delong's test showed that in training group,there was no significant difference in AUC between T2WI-ADC-PSAD model and T2WI model(P＞0.05),but there were significant differences between T2WI-ADC-PSAD model and other models(P＜0.05).In test group there were no significant differences in AUC between T2WI-ADC-PSAD model and other models(P＞0.05).The DCA showed that the T2WI-ADC-PSAD model provided a higher net benefit for clinical decision-making when the threshold probability was less than 97％.Conclusion BpMRI radiomics combined with PSAD can improve the diagnostic efficiency of low-grade and high-grade PCa,and guide the treatment decision of patients.

Tacrolimus (FK506) is a 23-membered macrolide with immunosuppressant activity that is widely used clinically for treating the rejection after organ transplantation. The research on tacrolimus production was mainly focused on biosynthesis methods, within which there are still some bottlenecks. This review summarizes the progress made in tacrolimus biosynthesis via modification of metabolic pathways and control of fermentation process, with the hope to address the technical bottlenecks for tacrolimus biosynthesis and improve tacrolimus production by fermentation engineering and metabolic engineering.
Tacrolimus ; Immunosuppressive Agents ; Fermentation ; Macrolides ; Anti-Bacterial Agents

L-methionine, also known as L-aminomethane, is one of the eight essential amino acids required by the human body and has important applications in the fields of feed, medicine, and food. In this study, an L-methionine high-yielding strain was constructed using a modular metabolic engineering strategy based on the M2 strain (Escherichia coli W3110 ΔIJAHFEBC/PAM) previously constructed in our laboratory. Firstly, the production of one-carbon module methyl donors was enhanced by overexpression of methylenetetrahydrofolate reductase (methylenetetrahydrofolate reductase, MetF) and screening of hydroxymethyltransferase (GlyA) from different sources, optimizing the one-carbon module. Subsequently, cysteamine lyase (hydroxymethyltransferase, MalY) and cysteine internal transporter gene (fliY) were overexpressed to improve the supply of L-homocysteine and L-cysteine, two precursors of the one-carbon module. The production of L-methionine in shake flask fermentation was increased from 2.8 g/L to 4.05 g/L, and up to 18.26 g/L in a 5 L fermenter. The results indicate that the one carbon module has a significant impact on the biosynthesis of L-methionine, and efficient biosynthesis of L-methionine can be achieved through optimizing the one carbon module. This study may facilitate further improvement of microbial fermentation production of L-methionine.
Humans ; Methionine ; Methylenetetrahydrofolate Reductase (NADPH2) ; Carbon ; Cysteine ; Escherichia coli/genetics* ; Hydroxymethyl and Formyl Transferases ; Carrier Proteins ; Escherichia coli Proteins

S-adenosyl-l-methionine (SAM) is ubiquitous in living organisms and plays important roles in transmethylation, transsulfuration and transamination in organisms. Due to its important physiological functions, production of SAM has attracted increasing attentions. Currently, researches on SAM production mainly focus on microbial fermentation, which is more cost-effective than that of the chemical synthesis and the enzyme catalysis, thus easier to achieve commercial production. With the rapid growth in SAM demand, interests in improving SAM production by developing SAM hyper-producing microorganisms aroused. The main strategies for improving SAM productivity of microorganisms include conventional breeding and metabolic engineering. This review summarizes the recent research progress in improving microbial SAM productivity to facilitate further improving SAM productivity. The bottlenecks in SAM biosynthesis and the solutions were also addressed.
S-Adenosylmethionine/metabolism* ; Plant Breeding ; Fermentation ; Metabolic Engineering

Major natural disasters seriously threaten human life and health. After earthquakes and other catastrophes, survivors are often trapped in the confined spaces caused by the collapse of ground and buildings, with relative separation from the outside world, restricted access, complex environment, and oncoming or ongoing unsafety, leading to the rescue extremely difficult. In order to save lives and improve the outcome more efficiently in the confined spaces after natural disasters, it is very important to standardize and reasonably apply the trauma assessment and first aid workflow. This study focuses on trauma assessment and first aid. From the aspects of trauma assessment, vital signs stabilization, hemostasis and bandaging, post-trauma anti-infection, and the transportation of patients, a trauma first aid work process suitable for a small space of a major natural disaster is formed, It is helpful to realize the immediate and efficient treatment of trauma in the confined spaces after natural catastrophes, to reduce the rate of death and disability and improve the outcome of patients.
Humans ; Disasters ; First Aid ; Confined Spaces ; Earthquakes

Objective:To compare postural reduction combined with percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) in the treatment of osteoporotic vertebral compression fractures (OVCFs).Methods:From January 2019 to January 2020,68 patients with OVCFs who met the inclusion and exclusion criteria in the Second Hospital of Tangshan Hebei Province were included in the observation study. A prospective randomized controlled study was used. The matched groups were divided into PVP combined group (adjust the overextension of the operating table by 20°-30°, if the posture reduction fails, pry the puncture needle on both sides in reverse according to the compression degree of the end plate before operation, and inject bone cement) and PKP group (do not adjust the operating table before operation, insert a balloon and expand on both sides after operation, and inject bone cement), with 34 cases in each group. The Cobb angle of the injured vertebrae was measured by taking the anterior and lateral X-ray film of the patient's lumbar spine before operation. The degree of pain and low back function were evaluated by visual analogue scale (VAS) and Oswetry disability index (ODI). The operation time and fluoroscopy times were recorded during the operation. On the second day after operation, the anterior and lateral X-ray of lumbar spine were taken to measure the Cobb angle of injured vertebrae. All patients were underwent computed tomography (CT) check the bone cement for leakage, record the VAS score, and record the ODI 3 months after operation to evaluate the patient's function. Follow up at the end of 12 months after operation to count the treatment cost and re-fracture of the patient. The data analysis and measurement data were compared by independent sample t-test between the two groups, paired sample t-test was used for intra-group comparison before and after operation. χ 2 test was used for counting data comparison between two groups. Results:All patients were followed up for 12 months. The operation time ((42.7±5.9) min), fluoroscopy times ((20.0±3.6) times) and treatment cost ((19 153±601) yuan) in the PVP combined group were better than those in the PKP Group ((67.4±7.3) min, (30.1±5.9) times, (27 496±669) yuan), and the difference was statistically significant ( t values were 15.39, 8.46, 54.12; all P<0.001). Cobb angle: Postoperative Cobb angle of injured vertebrae in the two groups (PVP combined group (10.7±4.5)°) and (PKP group (13.4±3.8)°) decreased compared with preoperative (PVP combined group (17.0±5.1)°) and (PKP group (16.7±5.1)°) ( t values were 10.61, 5.61; all P=0.001), and PVP combined group recovered better than PKP group, with statistically significant difference ( t=2.70, P=0.009). VAS score: Postoperative (PVP combined group (3.9±1.5) points) and (PKP group (4.1±1.6) points) was lower than preoperative the scores of (PVP combined group (6.9±1.1) points) and (PKP group (7.1±0.9) points), and the difference was statistically significant ( t values were 8.63, 8.88; all P=0.001). There was no significant difference in VAS scores between the two groups ( t=0.48, P=0.630). ODI scores: The scores of (PVP combined group (0.315±0.068)) and (PKP group (0.319±0.077)) after operation were lower than preoperative (PVP combined group (0.574±0.066), (PKP group (0.553±0.075)), and the difference was statistically significant ( t values were 18.54, 14.16, all P=0.001). There was no significant difference in ODI between the two groups ( t=0.25, P=0.803). There was no statistical significance in the two groups of postoperative bone cement leakage (χ 2=0.22, P=0.642). In PVP combined group, 1 case was re-fractured due to trauma, and there was no re-fracture in PKP group. Conclusion:Postural reduction combined with percutaneous needle prying reduction of PVP and PKP can alleviate the pain, improve the postoperative function and restore kyphosis in patients with OVCFs. Postural reduction combined with needle prying reduction of PVP has more advantages in operation time, radiation injury to doctors and patients, treatment cost, and the effect of correcting deformity is more significant.

Objective:To explore the feasibility and advantages of planned initiation of extracorporeal membrane oxygenation(ECMO)prior to liver transplantation.Methods:From November 2017 to July 2021, clinical data were retrospectively reviewed for 3 liver transplantation recipients assisted by ECMO.There were such preoperative symptoms of right ventricular dysfunction as fatigue, chest tightness and palpitations.In the first case, right heart catheterization was not performed due to patient refusal; another two patients were screened by transthoracic Doppler echocardiography(TDE)and diagnosed through right heart catheterization as portopulmonary hypertension(POPH)and pulmonary hypertension.Results:Three recipients with pulmonary hypertension received catheterization in right femoral artery and vein.After freeing of diseased liver and before blocking inferior vena cava, V-A ECMO support was performed.The dose of heparin was adjusted according to activated clotting time(ACT)and perioperative vital signs remained stable.They were ventilated for 54, 12 and 62 hours and supported by ECMO for 27, 61 and 14 hours.All were smoothly discharged.During a mean follow-up period of 26(9-22)months, liver functions were normal.Conclusions:Patients with end-stage liver disease with pulmonary hypertension should undergo routine TDE examinations during waiting period before liver transplantation.Those with pulmonary hypertension should undergo further right heart catheterization to confirm the diagnosis and severity of the disease.Planned application of ECMO through multidisciplinary consultations can expand surgical indications for liver transplantation, maintain intraoperative hemodynamic stability and facilitate smooth liver transplantation and postoperative patient recovery.

The redox biosynthesis system has important applications in green biomanufacturing of chiral compounds. Formate dehydrogenase (FDH) catalyzes the oxidation of formate into carbon dioxide, which is associated with the reduction of NAD(P)⁺ into NAD(P)H. Due to this property, FDH is used as a crucial enzyme in the redox biosynthesis system for cofactor regeneration. Nevertheless, the application of natural FDH in industrial production is hampered by low catalytic efficiency, poor stability, and inefficient coenzyme utilization. This review summarized the structural characteristics and catalytic mechanism of FDH, as well as the advances in protein engineering of FDHs toward improved enzyme activity, catalytic efficiency, stability and coenzyme preference. The applications of using FDH as a coenzyme regeneration system for green biomanufacturing of chiral compounds were summarized.
Catalysis ; Coenzymes/metabolism* ; Formate Dehydrogenases/metabolism* ; NAD/metabolism* ; Protein Engineering

As a strategic emerging industry of China, the biotechnology industry develops rapidly in recent years, which significantly increased the demand for creative and capable talents. As a core curriculum of bioengineering specialty, biotechnology equipment plays an important role in fostering such talents. To address the problems in biotechnology equipment course teaching such as limited equipment availability, limited engineering practice, and lack of learning motivations, curriculum reform and optimization were performed based on curriculum resource development, virtual reality-physical combined engineering training, and boosting learning motivations. The optimized teaching contents focus on fostering morality, intelligence, and creative practice abilities by connecting new requirements of social development, introducing new progress in biotechnology research, as well as new practices in research and development (R & D). Measures such as teaching methods innovation, assessment and evaluation methods optimization, cutting-edge R & D progress, diverse resources integration, and online-offline combined teaching, were developed to boost the learning motivation and foster the innovation competence of students. By above exploration and practice, the practice and innovation competence of students were significantly enhanced.
Humans ; Students ; Learning ; Curriculum ; Bioengineering ; Biomedical Engineering