Cellulose synthase (CesA), one of the key enzymes in the biosynthesis of cellulose in plants, plays an important role in plant growth and plant resistance. In this study, a total of 21 AsCesA genes from Aquilaria sinensis were systematically identified and the physico-chemical characteristics were analyzed based on genome database and bioinformatical methods. The phylogenetic tree was constructed and the gene location on chromosome, cis-acting elements in the 2 000 basepairs upstream regulatory regions and conservative motifs were analyzed. The AsCesA proteins were mainly located on the plasma membrane. The number of amino acids of the proteins ranged from 390 to 1 261. The isoelectric point distributed from 5.67 to 8.86. All of the 21 AsCesA proteins possessed the transmembrane domains, the number of which was from 6 to 8. The genes were classified into 3 groups according to the phylogenetic relationship. Obvious differences were observed in motif composition in genes from different groups. However, motif2, motif6, motif7 and motif10 were observed in all of AsCesA proteins. Analysis of cis-acting elements indicated that AsCesA genes family has cis-acting elements related to plant hormones, abiotic stresses, and biological processes. Seven AsCesA genes with differential expression were selected according to the calli transcriptome data induced by NaCl at different times and their expression levels under different abiotic stresses were analyzed by quantitative real-time PCR. The results indicated that salt, low temperature, drought, and heavy metal stresses could affect the expression level of AsCesA genes, and the abundance of AsCesA1, AsCesA3 and AsCesA20 showed a significant change, implying their potential important roles to the abiotic stresses. The accumulation pattern of cellulose content under different abiotic stresses was similar to the expression trend of AsCesA genes. Our results provide valuable insights into the role of cellulose synthase in A.sinensis in plant defense.

Objective:To explore the effects of early enteral nutrition (EEN) management based on severe feeding process on nutritional status and gastrointestinal tolerance in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) after mechanical ventilation.Methods:In the quasi trial study, 30 patients with AECOPD undergoing mechanical ventilation between January 2020 and June 2021 and other 30 patients undergoing mechanical ventilation between July 2021 and December 2022 in Liu′an Hospital of Traditional Chinese Medicine were enrolled as control group and observation group, respectively. The control group was given routine EEN intervention, while the observation group was given EEN management based on severe feeding process. The intakes of calorie and proteins within 7 d after intervention were compared between the two groups. The gastrointestinal tolerance and prognosis were recorded. The levels of hemoglobin, albumin and prealbumin were compared between the two groups before intervention and at 7 d after intervention.Results:In the control group, there were 17 males and 13 females, with age of (66.37±6.09) years old. In the observation group, there were 16 males and 14 females, with age of (65.49 ± 6.35) years old. After intervention, intakes of calorie and proteins within 7 d in the observation group were (3 513.62 ± 94.56) kJ/d and (51.06 ± 5.03) g/d, more than those in the control group (2 859.41 ± 87.23) kJ/d, (36.78 ± 3.14) g/d, the differences were statistically significant ( t=27.85, 13.19, both P<0.05). There were no significant difference in the levels of hemoglobin, albumin and prealbumin before intervention between the two groups (all P>0.05). After intervention, levels of albumin, hemoglobin and prealbumin were (37.16 ± 3.42), (135.43 ± 12.64) g/L and (218.54 ± 15.38) mg/L in the observation group, higher than those in the control group (34.25 ± 4.01), (127.51 ± 11.12) g/L and (201.33 ± 15.04) mg/L, the differences were statistically significant ( t=3.02, 2.58, 4.38, all P<0.05). The incidence rates of reflux aspiration and enteral nutrition interruption in the observation group were 6.67% (2/30), 6.67% (2/30), lower than those in the control group 26.67% (8/30), 30.00% (9/30), the differences were statistically significant ( χ2=4.32, 5.46, both P<0.05). The mechanical ventilation time and stay time in ICU in the observation group were (7.62 ± 1.35), (11.17 ± 2.04) d, shorter than those in the control group (8.57 ± 1.01), (12.83 ± 2.19) d, the differences were statistically significant ( t=3.09, 3.04, both P<0.05). Conclusions:EEN management based on severe feeding process can improve nutritional status, gastrointestinal tolerance and prognosis of AECOPD patients after mechanical ventilation.

Cyclin-dependent kinase 5 (CDK5), a serine/threonine kinase, is one of the non-typical members of the CDKs family. CDK5 is mainly activated by non-cyclin activators p35 or p39 (as well as their respective fragments p25 and p29) to phosphorylate downstream substrates and regulate numerous neural and non-neural functions. Increasing evidence has confirmed that the overactivation of CDK5/p25 complex is closely related to neurodegenerative diseases, cancers, diabetes and inflammation. Consequently, CDK5 has become an important target in multiple diseases treatment. Nevertheless, to date, no selective CDK5 inhibitors are currently in the clinical stage. On the other hand, pan-CDK inhibitors are limited in clinical trials, due to their poor clinical efficacy and toxic side effects caused by the extensive inhibition of other kinases. In view of this, selective CDK5 inhibitors are of great significance not only for elucidating its exact biological functions, but also exploring the possibility of CDK5 inhibitors as a safe and effective therapy. This paper provides a brief overview of the structure and function of CDK5 protein as well as its relationship with diseases. In addition, the structural types and binding modes of CDK5 inhibitors targeting ATP active sites are also highlighted. Finally, we summarize and prospect the strategies to improve the selectivity of CDK5 inhibitors.

Brasilicardin A, a diterpene glycoside isolated from pathogenic actinomycete Nocardia brasiliensis IFM 0406, has become a novel immunosuppressant candidate due to its significant immunosuppressive activity, low toxicity and unique mechanism of action. However, brasilicardin A and its analogues have become a research hotspot to the development of this promising immunosuppressant because of the low-yield production in the natural pathogenic producer and the synthetically challenging skeleton. According to the reported biosynthetic pathway of brasilicardin A, the function of involved diterpene synthase was analyzed by bioinformatics. Then the genes bra1-5 that synthesize the brasilicardin A skeleton were directionally amplified from the pathogenic strain N. brasiliensis IFM 0406, and heterologous expression was achieved successfully in Streptomyces albus R1. The compounds were isolated and purified by using various column chromatographies including silica gel column chromatography and semi-preparative HPLC. Six new brasilicardins were established and named brasilicardin H-M. The activity of brasilicardins was screened using lipopolysaccharide (LPS)-activated mouse primary macrophage inflammation model. Brasilicardin H-M exhibited good inhibitory activity on nitric oxide (NO) release with IC₅₀ values of 28.24 ± 3.70, 37.44 ± 2.00, 39.85 ± 4.02, 26.77 ± 4.40, 65.25 ± 1.48 and 15.24 ± 2.72 μmol·L^-1, respectively (indomethacin as the positive control with IC₅₀ value of 34.28 ± 4.10 μmol·L^-1). The results indicated that six compounds had potential anti-inflammatory activity. This study laid a foundation for the elucidation of the brasilicardin A biosynthetic pathway and evaluation of the structure-activity relationship as well as new drug developments.

Objective To explore root cause of poor prognosis after successful endovascular treatment(EVT)in patients with acute ischemic stroke with large vascular occlusion(AIS-LVO)of anterior circulation.Methods Patients with AIS-LOV of anterior circulation who received successful EVT(postoperative modified thrombolysis incerebral infarction[mTICI]grade≥2b)were retrospectively and continuously collected in the Department of Neurology of Bozhou People's Hospital from January 2022 to March 2024.The baseline and clinical data of the patients were collected,including gender,age,vascular risk factors(hypertension,diabetes,coronary heart disease,hyperlipidemia,valvular heart disease,atrial fibrillation,smoking,and alcohol consumption),prior stroke or transient ischemic attack,baseline blood pressure,baseline National Institutes of Health Stroke scale(NIHSS)score,laboratory test indicators(pre-operative C-reactive protein and D-dimer,post-operative fasting blood glucose,lipid levels,homocysteine,etc).Meanwhile,the data of perioperative indicators was collected,including the time from onset to admission,the time from admission to puncture,the time from puncture to revascularization,the time from onset to puncture,the time from onset to revascularization,remedial measures(balloon dilation,stent placement,arterial thrombolysis)during the surgery or not,using tirofiban or not,postoperative complications(stroke-related pneumonia,stress ulcers,deep vein thrombosis,acute heart failure or renal failure,etc)or not.The patient's medical history and imaging data were collected,and these indicators were defined and collected,including Alberta stroke program early CT score(ASPECTS),location of occlusion(C1 segment of the internal carotid artery,C2 segment to C7 segment of the internal carotid artery,M1 segment of the middle cerebral artery),and the trial of org 10172 in acute stroke treatment(TOAST)classification and a postoperative transformation of cerebral infarction after ischemic stroke and symptomatic intracranial hemorrhage or not.According to the modified Rankin scale(mRS)score at 90 d after surgery,all patients were divided into poor prognosis group(mRS score≥ 3)and good prognosis group(mRS score≤2).The baseline and clinical data of two groups were compared using univariate analysis.Variables with P＜0.1 in the univariate analysis were selected as independent variables,and the poor prognosis was used as the dependent variable.Further,multivariate Logistic regression analysis was performed to identify the influencing factors of poor prognosis after EVT.Results Finally,a total of 192 patients with AIS-LVO of anterior circulation who received successful revascularization were included in this study.There were 101 male patients and 91 female patients.The poor prognosis group had 102 cases and the good prognosis group had 90 cases.Univariate analysis showed that the poor prognosis group had statistically significant differences with the good prognosis group in terms of age(Z=-3.088,P=0.002)and age distribution(x2=13.457,P=0.001),fasting blood glucose(Z=-3.347,P=0.001),baseline NIHSS score(Z=-4.469,P＜0.01),location of occlusion(x2=10.488,P=0.005),transformation of hemorrhage after ischemic stroke(x2=16.943,P＜0.01),and symptomatic intracranial hemorrhage(X2=25.449,P＜0.01),and the baseline ASPECTS of the poor prognosis group was significantly lower than that of the good prognosis group(Z=-4.547,P＜0.01).There were no significant differences in other baseline and clinical data(all P＞0.05).Further multivariate Logistic regression analysis showed that age＞80 years(OR,3.224,95％CI 1.033-10.058,P=0.044),baseline NIHSS score(OR,1.102,95％CI 1.013-1.199,P=0.023),baseline ASPECTS(OR,0.375,95％CI 0.212-0.665,P=0.001),and symptomatic intracranial hemorrhage(OR,7.127,95％CI 1.296-39.203,P=0.024)were independent influencing factors of poor prognosis.Conclusion The independent factors of 90 d poor prognosis after successful EVT in patients with AIS-LVO of anterior circulation are age＞80 years,baseline NIHSS score,baseline ASPECTS,and symptomatic intracranial hemorrhage.

Objective To study the effect of ligustilide regulating stimulator of interferon genes(STING)/Tank-binding kinase 1(TBK1)/nuclear factor-κB(NF-κB)signaling on lung tissue damage in mouse model of lipopolysaccharide(LPS)-induced acute lung injury.Methods Mice were divided into control group,model group(LPS lung injury mouse model),experimental-L group(LPS lung injury mouse model,20 mg·kg-1 ligustilide treatment),experimental-M group(LPS lung injury mouse model,60 mg·kg-1 ligustilide treatment),experimental-H group(LPS lung injury mouse model,100 mg·kg-1 ligustilide treatment),DMXAA group(LPS lung injury mouse model,100 mg·kg-1 ligustilide,STING signal agonist DMXAA treatment),Western blot was used to detect the STING,p-TBK1,TBK1,p-p65,p65 protein expression,bicinchoninic acid(BCA)method was used to detect total protein in bronchoalveolar lavage fluid,Swiss Giemsa staining was used to count the number of inflammatory cells in bronchoalveolar lavage fluid,the content of tumor necrosis factor-alpha(TNF-α)was detected by enzyme-linked immunosorbent assay(ELISA),and the content of malondialdehyde(MDA)in lung tissue was detected by thiobarbituric acid method.Results The relative expression levels of STING protein in the control,model,experimental-L,experimental-M,experimental-H groups,and DMXAA group were 0.13±0.03,0.46±0.02,0.36±0.04,0.27±0.02,0.20±0.03 and 0.38±0.04,p-TBK1/TBK1 protein ratios were 0.15±0.02,0.56±0.02,0.43±0.01,0.31±0.01,0.20±0.02 and 0.30±0.02,p-p65/p65 protein ratios were 0.24±0.01,0.66±0.03,0.41±0.02,0.32±0.04,0.22±0.02 and 0.42±0.03,the total protein contents in bronchoalveolar lavage fluid were(173.79±15.44),(304.21±19.29),(243.59±17.60),(212.51±18.24),(187.11±11.59)and(241.69±30.12)μg·mL-1;the number of macrophages were(0.11±0.02)×108,(0.94±0.09)×108,(0.72±0.08)×108,(0.57±0.04)×108,(0.41±0.04)×108 and(0.71±0.08)×108 cell·mL-1.The difference of the above indicators between model group and control group were all statistically significant(all P＜0.05);the difference between experimental-L,experimental-M,experimental-H groups and model group were all statistically significant(all P＜0.05);there were statistically significant difference between DMXAA group and experimental-H group(all P＜0.05).Conclusion Ligustolide inhibits STING/TBK1/NF-κB signaling to alleviate lung tissue damage in a mouse model of LPS-induced acute lung injury.

Dihydroflavonol 4-reductase (DFR) plays an essential role in the biosynthesis of anthocyanin and regulation of plant flower color. Based on the transcriptome data of Cistanche tubulosa (Schenk) Wight, a full-length cDNA sequence of CtDFR gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR). CtDFR contains an open reading frame (ORF) of 1 263 bp which encodes 420 amino acids with a predicted molecular weight of 47.5 kDa. The sequence analysis showed that CtDFR contains a nicotinamide adenine dinucleotide phosphate (NADPH) binding domain and a specific substrate binding domain. The expression analysis indicated that CtDFR was highly expressed in red and purple flowers, and the relative expression levels were 4.04 and 19.37 times higher than those of white flowers, respectively. The recombinant CtDFR protein was expressed in E.coli BL21 (DE3) using vector pET-28a-CtDFR and was purified. In vitro enzyme activity analysis, CtDFR could reduce three types of dihydroflavonols including dihydrokaempferol, dihydroquercetin, and dihydromyricetin to leucopelargonidin, leucocyanidin and leucodelphinidin. Subcellular localization analysis showed that CtDFR was mainly localized in the cytoplasm. These results demonstrate that CtDFR plays an important role in regulation of flower color in C. tubulosa and make a valuable contribution for the further investigation on the regulation mechanism of C. tubulosa (Schenk) Wight flower color.

As a biocatalyst, enzyme has the advantages of high catalytic efficiency, strong reaction selectivity, specific target products, mild reaction conditions, and environmental friendliness, and serves as an important tool for the synthesis of complex organic molecules. With the continuous development of gene sequencing technology, molecular biology, genetic manipulation, and other technologies, the diversity of enzymes increases steadily and the reactions that can be catalyzed are also gradually diversified. In the process of enzyme-catalyzed synthesis, the majority of common enzymatic reactions can be achieved by single enzyme catalysis, while many complex reactions often require the participation of two or more enzymes. Therefore, the combination of multiple enzymes together to construct the multi-enzyme cascade reactions has become a research hotspot in the field of biochemistry. Nowadays, the biosynthetic pathways of more natural products with complex structures have been clarified, and secondary metabolic enzymes with novel catalytic activities have been identified, discovered, and combined in enzymatic synthesis of natural/unnatural molecules with diverse structures. This study summarized a series of examples of multi-enzyme-catalyzed cascades and highlighted the application of cascade catalysis methods in the synthesis of carbohydrates, nucleosides, flavonoids, terpenes, alkaloids, and chiral molecules. Furthermore, the existing problems and solutions of multi-enzyme-catalyzed cascade method were discussed, and the future development direction was prospected.
Biological Products/chemistry* ; Catalysis ; Alkaloids ; Biocatalysis

Objective To explore the effect of improving cleaning and disinfection methods on the prevention,con-trol and disinfection of carbapenem-resistant Enterobacterales(CRE)in burn plastic surgery ward.Methods 297 patients who admitted to the department of burn plastic surgery in a hospital from February 1 to August 31,2021 were selected as the control group,and 210 patients who admitted to the hospital from September 1,2021 to Febru-ary 28,2022 after cleaning and disinfection methods improved were selected as the intervention group.Detection rate of CRE from patients,incidence of healthcare-associated infection(HAI)with CRE,and detection rate of envi-ronmental CRE before and after intervention were statistically analyzed and compared.Results The incidence of HAI and detection rate of CRE from patients in the intervention group were 0.95％and 0,respectively,lower than 4.04％and 2.02％in the control group(both P＜0.05).Compared to the control group,qualified rates of detec-tion of air and surface microbiology,adenosine triphosphate(ATP)bioflorescence and fluorescence labeling in the intervention group were all higher(x2=5.52,13.08,6.66,and 15.01,respectively,all P＜0.05).Conclusion Improving cleaning and disinfection method can reduce the incidence of HAI and the detection rate of CRE in burn wards,improve the surface cleanliness of environmental objects,as well as the effectiveness of HAI prevention and control.

OBJECTIVE: To observe the effect of needle-knife on the chondrocyte apoptosis of knee joint in rabbits with knee osteoarthritis (KOA) based on the CircSERPINE2-miR-1271-5P-E26 specific transformation-related gene (ERG) axis, and to explore the mechanism of needle-knife for KOA. METHODS: Thirty-six New Zealand white rabbits were randomly divided into a normal group, a model group, a needle-knife group and a sham needle-knife group, 9 rabbits in each group. The rabbits in the model group, the needle-knife group and the sham needle-knife group were treated with modified Videman method to prepare KOA model. After successful modeling, the rabbits in the needle-knife group were treated with needle-knife at cord adhesion and nodules near quadriceps femoris tendon and internal and external collateral ligament on the affected knee joint; the rabbits in the sham needle-knife group were treated with sham needle-knife baside the needle insertion point of the needle-knife group (needle-knife was only inserted, without any operation). The treatment was given once a week, 3 times in total. The Lequesne MG behavioral score was used to evaluate the knee joint damage in each group before and after intervention. After intervention, HE staining and transmission electron microscopy were used to observe the cartilage tissue morphology and ultrastructure of chondrocytes in the knee joint in each group; TUNEL method was used to detect the level of chondrocyte apoptosis in the knee joint; real-time fluorescence quantitative PCR was used to detect the expression of CircSERPINE2, miR-1271-5P and ERG mRNA in knee cartilage tissue in each group. RESULTS: After intervention, compared with the normal group, the Lequesne MG behavioral score in the model group was increased (P<0.01). Compared with the model group and the sham needle-knife group, the Lequesne MG behavioral score in the needle-knife group was decreased (P<0.01). In the model group and the sham needle-knife group, the number of chondrocytes and organelles was decreased, the cell nucleus was shrunk, mitochondria was swelling or disappeared; in the needle-knife group, the number of chondrocytes and organelles was increased, the cell nucleus was not obviously shrunk and the mitochondria was not obviously swelling. Compared with the normal group, the level of chondrocyte apoptosis in the model group was increased (P<0.01); compared with the model group and the sham needle-knife group, the level of chondrocyte apoptosis in the needle-knife group was decreased (P<0.01, P<0.05). Compared with the normal group, the expression of CircSERPINE2 and ERG mRNA in the model group was decreased (P<0.01), and the expression of miR-1271-5P mRNA was increased (P<0.01); compared with the model group and the sham needle-knife group, the expression of CircSERPINE2 and ERG mRNA in the needle-knife group was increased (P<0.01), and the expression of miR-1271-5P mRNA was decreased (P<0.01). CONCLUSION Needle-knife could reduce the knee joint damage and chondrocyte apoptosis in KOA rabbits, which may be related to up-regulating the expression of CircSERPINE2 and ERG mRNA, and inhibiting the expression of miR-1271-5P mRNA.
Rabbits ; Animals ; Osteoarthritis, Knee/metabolism* ; Chondrocytes/metabolism* ; Knee Joint/surgery* ; Apoptosis ; MicroRNAs/genetics*