ObjectiveTo prepare a recombinant PETase with a dual-catalytic-triad and to evaluate its efficiency in the biodegradation of polyethylene terephthalate （PET）. MethodsBased on the crystal structure of wild-type PETase， point mutations （T88H/L117D） were introduced via site-directed mutagenesis. The recombinant protein was prepared using prokaryotic expression and chromatography purification techniques. The enzymatic hydrolysis of the mutant PETase was assessed by relatively quantifying the products mono （2-hydroxyethyl） terephthalate （MHET） and terephthalic acid （TPA）. ResultsBoth wild-type and mutant PETases accumulated as inclusion bodies， accounting for approximately 20% of the total bacterial protein. After solubilization in urea， the proteins were eluted at 300 mmol/L imidazole during affinity chromatography purification， with concentrations of 1.824 and 1.833 mg/mL and purities of 83.11% and 84.32%， respectively. Subsequent anion-exchange chromatography yielded highly pure enzymes in the 200 mmol/L NaCl fraction： 2.776 mg/mL （96.86% purity） for the wild type and 1.967 mg/mL （95.13% purity） for the mutant. Following refolding， the final concentrations were 0.484 mg/mL for the wild type and 0.991 mg/mL for the mutant. Hydrolysis assays revealed that the mutant released MHET and TPA at （237.67±17.00）% and （197.33±12.01）% of the wild-type levels， respectively. ConclusionThe T88H/L117D dual-catalytic-triad PETase is successfully prepared and it significantly enhanced PET-degrading activity， thus， it′s a promising biocatalyst for PET bioremediation.

We reported a case of eosinophilic meningoencephalitis,presenting with symptoms such as depression,headache and memory decline.Physical examination revealed mental tension,slightly slurred speech,and cognitive decline.Laboratory tests indicated a significant increase in eosinophils in both peripheral blood and cerebrospinal fluid,suggesting eosinophilic meningoencephalitis.After treatment with glucocorticoids and cognitive function improvement measures,the patient's symptoms improved.Here,we summarized its clinical features,laboratory examination,diagnosis and treatment.Then,we discussed the pathogenesis,treatment and differential diagnosis of eosinophilia manifested as meningoencephalitis.In order to improve clinicians'understanding of eosinophilia complicated with meningoencephalitis and provide reference for clinical diagnosis and treatment of these diseases.

We reported a case of eosinophilic meningoencephalitis,presenting with symptoms such as depression,headache and memory decline.Physical examination revealed mental tension,slightly slurred speech,and cognitive decline.Laboratory tests indicated a significant increase in eosinophils in both peripheral blood and cerebrospinal fluid,suggesting eosinophilic meningoencephalitis.After treatment with glucocorticoids and cognitive function improvement measures,the patient's symptoms improved.Here,we summarized its clinical features,laboratory examination,diagnosis and treatment.Then,we discussed the pathogenesis,treatment and differential diagnosis of eosinophilia manifested as meningoencephalitis.In order to improve clinicians'understanding of eosinophilia complicated with meningoencephalitis and provide reference for clinical diagnosis and treatment of these diseases.

Objective To investigate the effect of platelet lysate on mitochondrial function of astrocytes after spinal cord injury.Methods Astrocytoma cells SVGp12 were incubated with different concentrations of H2O2 (100 μmol/L,400 μmol/L,1000 μmol/L,2000 μmol/L),and the OD value was determined by CCK-8 assay. The H2O2 concentration with a cell inhibition rate of about 50％ was selected for follow-up experiments. The cells were incubated with different concentrations of platelet lysate (the volume ratios of platelet lysate to culture medium were 1∶10,1∶40,1∶80,1∶160 and 1∶320) for 24 hours,and the OD value was determined,thereby selecting the optimal concentration of platelet lysate based on the cell survival rate. SVGp12 cells were divided into the normal group (cultured normally without special treat-ment),the model group (incubated with 1000 μmol/L of H2O2 for 3 hours),and the treatment group (incubated with 1000 μmol/L of H2O2 for 3 hours and then treated with platelet lysate at a volume ratio of 1∶80). The mitochondrial function was evaluated by mitochondrial activity staining,mitochondrial membrane potential detection,and mitochondrial permeability transition pore detection. Results After incubating cells with 1000 μmol/L of H2O2 for 3 hours,the cell inhibition rate was 48％,and the OD value decreased significantly (P<0.01),confirming the successful establishment of a model of astrocytes with spinal cord injury. After treated with different concentrations of platelet lysate,the OD values and cell survival rate of the cells were higher than those in the model group (P<0.05). The concentration of platelet lysate (volume ratio of 1∶80) with cell viability of 20％ was selected for follow-up experiments. Platelet lysate could improve the morphology of injured astrocytes. The mitochondrial activity of the treatment group was significantly higher than that of the model group,and the mitochondrial activity of the model group was significantly lower than that of the normal group,with statistically significant differences (P<0.05). The mitochondrial membrane potential in the model group was lower than that in the normal group,and the mitochondrial membrane potential in the treatment group was higher than that in the model group,with statistically significant differences (P<0.01). The mitochondrial permeability in the model group was greater than that in the normal group,and the mitochondrial permeability in the treatment group was smaller than that in the model group,with statistically significant differences (P<0.01).Conclusion Platelet lysate can improve the survival rate of astrocytes after spinal cord injury,enhance the mitochondrial activity of cells,and improve the mitochondrial membrane potential and mitochondrial permeability transition pore opening.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective To investigate the effect of platelet lysate on mitochondrial function of astrocytes after spinal cord injury.Methods Astrocytoma cells SVGp12 were incubated with different concentrations of H2O2 (100 μmol/L,400 μmol/L,1000 μmol/L,2000 μmol/L),and the OD value was determined by CCK-8 assay. The H2O2 concentration with a cell inhibition rate of about 50％ was selected for follow-up experiments. The cells were incubated with different concentrations of platelet lysate (the volume ratios of platelet lysate to culture medium were 1∶10,1∶40,1∶80,1∶160 and 1∶320) for 24 hours,and the OD value was determined,thereby selecting the optimal concentration of platelet lysate based on the cell survival rate. SVGp12 cells were divided into the normal group (cultured normally without special treat-ment),the model group (incubated with 1000 μmol/L of H2O2 for 3 hours),and the treatment group (incubated with 1000 μmol/L of H2O2 for 3 hours and then treated with platelet lysate at a volume ratio of 1∶80). The mitochondrial function was evaluated by mitochondrial activity staining,mitochondrial membrane potential detection,and mitochondrial permeability transition pore detection. Results After incubating cells with 1000 μmol/L of H2O2 for 3 hours,the cell inhibition rate was 48％,and the OD value decreased significantly (P<0.01),confirming the successful establishment of a model of astrocytes with spinal cord injury. After treated with different concentrations of platelet lysate,the OD values and cell survival rate of the cells were higher than those in the model group (P<0.05). The concentration of platelet lysate (volume ratio of 1∶80) with cell viability of 20％ was selected for follow-up experiments. Platelet lysate could improve the morphology of injured astrocytes. The mitochondrial activity of the treatment group was significantly higher than that of the model group,and the mitochondrial activity of the model group was significantly lower than that of the normal group,with statistically significant differences (P<0.05). The mitochondrial membrane potential in the model group was lower than that in the normal group,and the mitochondrial membrane potential in the treatment group was higher than that in the model group,with statistically significant differences (P<0.01). The mitochondrial permeability in the model group was greater than that in the normal group,and the mitochondrial permeability in the treatment group was smaller than that in the model group,with statistically significant differences (P<0.01).Conclusion Platelet lysate can improve the survival rate of astrocytes after spinal cord injury,enhance the mitochondrial activity of cells,and improve the mitochondrial membrane potential and mitochondrial permeability transition pore opening.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective To observe the effects of Zhengan Xifeng decoction on bile acid-Takeda G protein-coupled receptor 5(TGR5)/Ras homolog(Rho)/Rho-associated coiled-coil containing protein kinase(ROCK)/agouti-related protein(AgRP)signaling pathway in spontaneously hypertensive rats(SHR).Methods Seventy SHR were randomly divided into the model group,control group(gavage administration of 0.5 mg·kg-1·d-1 amlodipine besylate tablets),experimental-L,-M,-H group(gavage administration of 8.630 x 103,1.725 × 104,3.450 × 104 mg·kg-1·d-1 Zhengan Xifeng decoction).Fifteen Wistar-Kyoto rats served as the normal group.The model and normal groups were given the same amount of distilled water.Six groups were treated for 8 weeks with once a day.The bile acid profiles of jejunum contents and colon contents were detected by ultra-high-performance liquid chromatography-tandem mass spectrometry,and the expression levels of TGR5,Rho,ROCK and AgRP proteins in the hypothalamus were detected by Western blot.Results Compared with the normal,mode,control groups and experimental-L,-M,-H groups,the bile acid spectrum of jejunum contents was significantly changed,and 6 different bile acids were found,namely glycoursodeoxycholic acid,glycohyodeoxycholic acid,hyodeoxycholic acid,ursodeoxycholic acid,taurochenodeoxycholic acid and chenodeoxycholic acid.The bile acid spectrum of colon contents was significantly changed,and 4 different bile acids were found,namely deoxycholic acid,ursodeoxycholic acid,lithocholic acid and hyodeoxycholic acid.The relative expression levels of TGR5 protein in normal,model,control and experimental-M groups were 1.01±0.22,0.75±0.23,0.96±0.16 and 1.02±0.21;the relative expression levels of Rho protein were 0.81±0.14,0.58±0.19,0.83±0.27 and 0.94±0.24;the relative expression levels of ROCK protein were 0.99±0.27,0.65±0.13,1.07±0.22 and 1.06±0.24;the relative expression levels of AgRP protein were 0.98±0.33,1.46±0.45,0.83±0.25 and 0.95±0.33,respectively.Compared with the model group,the above indexes in the experimental-M group and the normal group showed significant differences(all P＜0.05).Conclusion Zhengan Xifeng decoction may suppress AgRP expression via bile acid-TGR5/Rho/ROCK/AgRP signaling pathway,to achieve the purpose of reducing blood pressure.

Objective To observe the effects of Zhengan Xifeng decoction on bile acid-Takeda G protein-coupled receptor 5(TGR5)/Ras homolog(Rho)/Rho-associated coiled-coil containing protein kinase(ROCK)/agouti-related protein(AgRP)signaling pathway in spontaneously hypertensive rats(SHR).Methods Seventy SHR were randomly divided into the model group,control group(gavage administration of 0.5 mg·kg-1·d-1 amlodipine besylate tablets),experimental-L,-M,-H group(gavage administration of 8.630 x 103,1.725 × 104,3.450 × 104 mg·kg-1·d-1 Zhengan Xifeng decoction).Fifteen Wistar-Kyoto rats served as the normal group.The model and normal groups were given the same amount of distilled water.Six groups were treated for 8 weeks with once a day.The bile acid profiles of jejunum contents and colon contents were detected by ultra-high-performance liquid chromatography-tandem mass spectrometry,and the expression levels of TGR5,Rho,ROCK and AgRP proteins in the hypothalamus were detected by Western blot.Results Compared with the normal,mode,control groups and experimental-L,-M,-H groups,the bile acid spectrum of jejunum contents was significantly changed,and 6 different bile acids were found,namely glycoursodeoxycholic acid,glycohyodeoxycholic acid,hyodeoxycholic acid,ursodeoxycholic acid,taurochenodeoxycholic acid and chenodeoxycholic acid.The bile acid spectrum of colon contents was significantly changed,and 4 different bile acids were found,namely deoxycholic acid,ursodeoxycholic acid,lithocholic acid and hyodeoxycholic acid.The relative expression levels of TGR5 protein in normal,model,control and experimental-M groups were 1.01±0.22,0.75±0.23,0.96±0.16 and 1.02±0.21;the relative expression levels of Rho protein were 0.81±0.14,0.58±0.19,0.83±0.27 and 0.94±0.24;the relative expression levels of ROCK protein were 0.99±0.27,0.65±0.13,1.07±0.22 and 1.06±0.24;the relative expression levels of AgRP protein were 0.98±0.33,1.46±0.45,0.83±0.25 and 0.95±0.33,respectively.Compared with the model group,the above indexes in the experimental-M group and the normal group showed significant differences(all P＜0.05).Conclusion Zhengan Xifeng decoction may suppress AgRP expression via bile acid-TGR5/Rho/ROCK/AgRP signaling pathway,to achieve the purpose of reducing blood pressure.

Objective:To analyze the effects of highdose methotrexate(HD-MTX)and lenalidomide as central nervous system(CNS)prophylaxis strategies in patients with diffuse large B-cell lymphoma(DLBCL).Methods:The data of DLBCL patients with high risk of CNS recurrence who were initially treated in Fujian Provincial Hospital and Fujian Cancer Hospital from January 2012 to June 2022 were analyzed retrospectively.The patients were divided into HD-MTX group and lenalidomide group according to different prophylaxis strategies.Each group was further divided into high-risk group and medium-risk group based on CNS-IPI score and/or testicular involvement.The CNS relapse-free survival(CRFS)rate,adverse effects,and the effects of different prophylaxis strategies on overall survival(OS)rate and progression-free survival(PFS)rate were evaluated in different groups and subgroups.Results:There were 200 patients enrolled in this study,80 cases in lenalidomide group and 120 cases in HD-MTX group.According to the delivery timing of prophylactic HD-MTX,the patients in HD-MTX group were further divided into two groups:80 cases at the end of induction chemotherapy and 40 cases during chemotherapy interval.At a median follow-up of 48(14-133)months,the 4-year CRFS rate,4-year PFS rate,and 4-year OS rate of the HD-MTX group was 93.6％,57.2％,and 68.8％,respectively,while that of the lenalidomide group was 90.4％,69.4％and 75.6％.There were no significant differences in 4-year CRFS rate,4-year PFS rate,and 4-year OS rate between HD-MTX group and lenalidomide group(all P＞0.05),but lenalidomide group showed a trend of improvement in PFS.Further subgroup analysis showed that there was no significant difference in 4-year CRFS rate between high-risk patients of the two groups(91.7％vs 83.4％,P＞0.05),while 4-year PFS rate showed difference(49.5％vs 64.2％,P＜0.05).A total of 248 cycles were collected for adverse reaction analysis in the HD-MTX group,and 25 cycles occurred neutropenia accompanied with infection(10.1％),while in lenalidomide group 240 cycles were collected in which 20 cycles occurred neutropenia accompanied with infection(8.3％).Both the two groups had no treatment-related deaths.Conclusion:Compared with HD-MTX,lenalidomide combined with immunochemotherapy can prevent CNS relapse,at the same time,improve prognosis,which is a safe and well tolerated central prophylaxis strategy.