ObjectiveSodium hyaluronate (HA) was used as the research object to modify it with phenolic acid in order to obtain the molecular structure with better antioxidant activity or even new activity. MethodsIn this study, 5 kinds of phenolic acid-sodium hyaluronate was prepared by free radical-mediated grafting method, and the grafts with the highest grafting degree were selected to optimize the synthesis conditions. Then, grafts structure and physicochemical properties were analyzed. The grafts were characterized by IR, UV, ¹H NMR, FESEM and TGA spectra. The in vitro antioxidant capacity of grafts was determined by the scavenging ability of DPPH·, ABTS⁺· and O^2-·. ResultsAmong 5 kinds of phenolic acid-sodium hyaluronate, the grafting rate of ferulic acid-sodium hyaluronate copolymer (FA-HA) was highest , which was chosen as experimental sample in the following tests. Firstly, the reaction conditions were investigated and the highest grafting rate was (16.59±0.31) mg/g at the optimal preparation conditions. Then, FA-HA structure and physicochemical properties were analyzed. Data from UV, IR, ¹H NMR analyses, TGA showed that FA were successfully grafted to HA. Compared with HA, the results of gel permeation chrematography (GPC) showed that the molecular mass distribution ofFA-HA copolymer decreased from 34.4 to 31.5 ku, but the uniformity of molecular distribution was improved. FESEM results showed that the structure of copolymer exhibited a closely connected lamellar structure with a relatively smooth surface. TGA results showed that thermal stability of FA-HA had a little decline. The antioxidant performance in vitro results showed that, during 0.25-10 g/L, FA-HA can eliminate (83.76±4.86)% DPPH·, (76.95±5.06)% ABTS⁺· and (83.08±2.51)% O^2-· respectively at 10 g/L. which were higher than that of native HA and FA. ConclusionFA and HA were successfully grafted together by free radical grafting, and the grafted FA-HA had better antioxidant activity in vitro, which provided a theoretical basis for further research and development of phenolic acid-HA grafts.

Sodium-glucose linked transporter 2 inhibitors（SGLT2i） are novel oral hypoglycaemic agents and are widely used for hypoglycemic therapy in patients with type 2 diabetes mellitus， but differences in the genetic backgrounds of patients often lead to variable responsiveness to drug therapy. By summarizing the pharmacogenomic studies of SGLT2i， the article found that the genetic variants of UGT1A9， UGT2B4， SLC5A2， ABCB1， PNPLA3 and WFS1 may influence the pharmacokinetics of SGLT2i and the external hypoglycemic effects of SGLT2i in improving non-alcoholic fatty liver disease， weight loss and so on， but there is no clinical evidence that genetic polymorphisms affect the hypoglycemic efficacy of SGLT2i.

Objective To investigate the expression of serum Maresin 1 and pro-platelet basic protein(PPBP)in patients with diabetes nephropathy(DN)and their correlation with long-term prognosis.Methods A total of 83 patients with diabetes nephropathy admitted to Tangshan Central Hospital from May 2018 to May 2020 were selected as the diabetes nephropathy group.In the same period,60 patients with simple type 2 diabetes were selected as the diabetes group and 60 healthy people as the control group.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of serum Maresin1 and PPBP.Spearman correlation analysis was used to analyze the correlation between the levels of serum Maresin1 and PPBP and renal pathological damage.COX proportional hazard regression analysis was used to analyze the factors influencing the long-term poor prognosis of patients with diabetes nephropathy.ROC curve was used to evaluate the predictive value of serum Maresin1 and PPBP for the long-term poor prognosis.Results The levels of serum Maresin1(15.90±4.53 ng/ml,12.34±4.29 ng/ml,9.65±4.38 ng/ml)in the control group,diabetes group and diabetes nephropathy group were decreased in turn while the levels of serum PPBP(263.45±85.22 pg/ml,349.28±80.49 pg/ml,435.76±87.21 pg/ml)were increased in turn,and the differences were statistically significant(F=35.159,72.678,all P＜0.05).With the increase of IFTA score,interstitial inflammation score,and glomerular grading,serum Maresin1 level was decreased(F=25.838,25.187,9.751,all P＜0.05),while serum PPBP level was increased(F=56.513,92.702,58.137,all P＜0.05),and the differences were statistically significant,respectively.Serum Maresin1 was negatively correlated with IFTA score,interstitial inflammation score,and glomerular grading(r=-0.637,-0.581,-0.594,all P＜0.05),while serum PPBP was positively correlated with IFTA score,interstitial inflammation score,and glomerular grading(r=0.659,0.664,0.608,all P＜0.05),with significant differences.The course of diabetes nephropathy(HR=1.135,95％CI:1.012～1.370),24-hour urinary protein(HR=1.087,95％CI:1.016～1.164),PPBP(HR=1.208,95％CI:1.119～1.365),and IFTA score(HR=1.139,95％CI:1.024～1.219),interstitial inflammation score(HR=1.122,95％CI:1.006～1.249)and glomerular grading(HR=1.139,95％CI:1.052～1.273)were independent risk factors for long-term prognosis of diabetes nephropathy patients,while eGFR(HR=0.934,95％CI:0.892～0.993)and Maresin1(HR=0.903,95％CI:0.816～0.982)were protective factors for long-term prognosis(all P＜0.05).The areas under the curve(AUC)of serum Maresin1,PPBP and two indicators combined to predict the long-term poor prognosis of patients with diabetes nephropathy were 0.781,0.777 and 0.901,respectively.The AUC of two indicators combined was higher than that,and the differences were significant(Z=3.049,3.258,all P＜0.05).Conclusion In patients with diabetes nephropathy,serum Maresin1 was decreased and PPBP was increased,and the two indexes were closely related to the degree of renal injury.The combined test could effectively predict the long-term poor prognosis of patients.

Objective: To investigate the role of miR-142-3p in alleviation of house dust mite induced allergic airway inflammation among children, so as to provide insights into unraveling the pathogenesis of allergic airway inflammation. Methods: Serum samples were collected from 15 patients with house dust mite induced allergic asthma and 15 healthy children in Jiangnan University Medical Center from September to November 2022, and serum miR-142-3p expression was quantified using a fluorescent quantitative real time PCR (qPCR) assay. The levels of interleukin 6 (IL 6) and tumor necrosis factor α (TNF α) were measured in the cell culture supernatant using enzyme linked immunosorbent assay (ELISA), and the expression of high mobility group box 1 (HMGB1) was detected at transcriptional and translational lvels using qPCR and Western blotting assays. The negative regulation of the HMGB1 gene by miR 142 3p was identified using a dual luciferase gene reporter assay, and the expression of downstream regulatory proteins was determined in human normal lung epithelial cells (BEAS 2B) cells transfected with miR 142 3p using Western blotting. In addition, female C57BL/6 mice at ages of 6-8 weeks were randomly assigned to the phosphate buffer saline (PBS) group, house dust mite sensitized airway inflammation group and house dust mite sensitized airway inflammation + miR 142 3p intervention group. Mouse airway inflammation was evaluated using hematoxylin eosin staining, and the expression of inflammatory cells and inflammatory cytokines were detected in mouse bronchoalveolar lavage fluid (BALF) using Giemsa staining and ELISA. Results: Lower serum miR-142-3p expression was quantified among children with house dust mite induced allergic asthma than among healthy controls (1.33±0.21 vs. 4.74±0.62, t=5.22, P <0.05). Stimulation with dermatophagoides farinae extract (DFE) resulted in a reduction in miR-142-3p expression in BEAS-2B cells (0.82±0.25), while transfection with miR-142-3p mimics resulted in a rise in miR-142-3p expression in BEAS-2B cells (0.55±0.14)( t=3.31, 3.94, P <0.05). Pre treatment with miR-142-3p reduced the expression of IL 6(2.25±0.46)and TNF α(6.58±1.95) ( t=4.86, 3.38, P <0.05) in BEAS 2B cells stimulated with DFE, and treatment with miR-142-3p mimics resulted in a reduction in TLR4 and NF-κB expression in BEAS-2B cells via negative regulation of the HMGB1 expression. In addition, treatment with miR-142-3p was found to alleviate inflammatory cell infiltration in lung tissues of house dust mite sensitized mice, and results in a reduction in interleukin 4 (IL-4)[(107.60±10.43)pg/mL], interleukin 5 (IL 5)[(95.78±13.14)pg/mL] and HMGB1[(2.52±0.87)pg/mL] expression in BALF ( t=10.32, 7.29, 2.90, P <0.05). Conclusion miR-142-3p alleviates house dust mite induced allergic airway inflammation among children via negative regulation of the HMGB1/TLR4/NF-κB pathway.

L-homophenylalanine (L-HPA) is an important non-natural amino acid that has been used as a key intermediate for the synthesis of Puli drugs for the treatment of hypertension. At present, L-HPA is synthesized using chemical methods, which has the disadvantages of expensive raw materials, tedious steps and serious pollution. Therefore, researchers have conducted in-depth research on the enzymatic production of L-HPA. This review summarizes the research progress on the enzymatic synthesis of L-HPA, including the dehydrogenase process, the transaminase process, the hydantoinase process, and the decarboxylase process, with the hope to facilitate the industrial production of L-HPA.
Amino Acids ; Environmental Pollution ; Industry ; Protein Biosynthesis

Succinic acid is an important C4 platform chemical that is widely used in food, chemical, medicine sectors. The bottleneck of fermentative production of succinic acid by engineered Escherichia coli is the imbalance of intracellular cofactors, which often leads to accumulation of by-products, lower yield and low productivity. Stoichiometric analysis indicated that an efficient production of succinic acid by E. coli FMME-N-26 under micro-aeration conditions might be achieved when the TCA cycle provides enough ATP and NADH for the r-TCA pathway. In order to promote succinic acid production, a serial of metabolic engineering strategies include reducing ATP consumption, strengthening ATP synthesis, blocking NADH competitive pathway and constructing NADH complementary pathway were developed. As result, an engineered E. coli FW-17 capable of producing 139.52 g/L succinic acid and 1.40 g/L acetic acid in 5 L fermenter, which were 17.81% higher and 67.59% lower than that of the control strain, was developed. Further scale-up experiments were carried out in a 1 000 L fermenter, and the titer of succinic acid and acetic acid were 140.2 g/L and 1.38 g/L, respectively.
Escherichia coli/genetics* ; NAD ; Succinic Acid ; Acetic Acid ; Adenosine Triphosphate

As a branched chain amino acid, L-valine is widely used in the medicine and feed sectors. In this study, a microbial cell factory for efficient production of L-valine was constructed by combining various metabolic engineering strategies. First, precursor supply for L-valine biosynthesis was enhanced by strengthening the glycolysis pathway and weakening the metabolic pathway of by-products. Subsequently, the key enzyme in the L-valine synthesis pathway, acetylhydroxylate synthase, was engineered by site-directed mutation to relieve the feedback inhibition of the engineered strain. Moreover, promoter engineering was used to optimize the gene expression level of key enzymes in L-valine biosynthetic pathway. Furthermore, cofactor engineering was adopted to change the cofactor preference of acetohydroxyacid isomeroreductase and branched-chain amino acid aminotransferase from NADPH to NADH. The engineered strain C. glutamicum K020 showed a significant increase in L-valine titer, yield and productivity in 5 L fed-batch bioreactor, up to 110 g/L, 0.51 g/g and 2.29 g/(L‧h), respectively.
Valine ; Corynebacterium glutamicum/genetics* ; Metabolic Engineering ; Amino Acids, Branched-Chain ; Bioreactors

OBJECTIVE: To explore clinical effect of repairing anterior talofibular ligament with knot-free anchors under total ankle arthroscopy in treating chronic lateral ankle instability. METHODS: From April 2018 to August 2021, 24 patients with chronic lateral ankle instability were treated with knot-free anchors under total ankle arthroscopy to repair anterior talofibular ligament, including 16 males and 8 females, aged from 22 to 42 years old with an average of(28.6±5.8) years old;the time from injury to opertaion ranged from 6 to 10 months with an average of(7.7±1.3) months. Preoperative and postoperative American Orhopaedic Foot and Ankle Society (AOFAS) score, visual analogue scale (VAS), talar tilt, anterior talar translation(ATT) were recorded and compared. RESULTS: All patients were followed up from 10 to 12 months with an average of (10.2±1.14) months. Incision were healed at stageⅠ, and no infection, nerve injury and lateral ankle instability occurred. AOFAS score improved from(52.79±8.96) before opertaion to (93.00± 4.01) at 6 months after operation, 23 patients got excellent result and 1 good;VAS decreased from (5.50±0.98) before opertaion to (1.04±0.80) at 6 months after operation(P<0.05);talar tilt decreased from(9.16±2.09)° to (3.10±1.72)° at 3 months after operation(P<0.05);ATT decreased from(8.80±2.55) mm to (2.98±1.97) mm at 3 months after operation(P<0.05). Twenty-four patients drawer test and varus-valgus rotation wer negative. CONCLUSION Repairing anterior talofibular ligament with knot-free anchors under total ankle arthroscopy for the treatment of chronic lateral ankle instability has advantages of less trauma, less complications safe and reliable, and good recovery of ankle joint function.
Female ; Male ; Humans ; Young Adult ; Adult ; Ankle Joint/surgery* ; Ankle ; Arthroscopy ; Lateral Ligament, Ankle/surgery* ; Joint Instability/surgery*

Chronic pancreatitis (CP) is a progressive and irreversible fibroinflammatory disorder, accompanied by pancreatic exocrine insufficiency and dysregulated gut microbiota. Recently, accumulating evidence has supported a correlation between gut dysbiosis and CP development. However, whether gut microbiota dysbiosis contributes to CP pathogenesis remains unclear. Herein, an experimental CP was induced by repeated high-dose caerulein injections. The broad-spectrum antibiotics (ABX) and ABX targeting Gram-positive (G⁺) or Gram-negative bacteria (G^-) were applied to explore the specific roles of these bacteria. Gut dysbiosis was observed in both mice and in CP patients, which was accompanied by a sharply reduced abundance for short-chain fatty acids (SCFAs)-producers, especially G⁺ bacteria. Broad-spectrum ABX exacerbated the severity of CP, as evidenced by aggravated pancreatic fibrosis and gut dysbiosis, especially the depletion of SCFAs-producing G⁺ bacteria. Additionally, depletion of SCFAs-producing G⁺ bacteria rather than G^- bacteria intensified CP progression independent of TLR4, which was attenuated by supplementation with exogenous SCFAs. Finally, SCFAs modulated pancreatic fibrosis through inhibition of macrophage infiltration and M2 phenotype switching. The study supports a critical role for SCFAs-producing G⁺ bacteria in CP. Therefore, modulation of dietary-derived SCFAs or G⁺ SCFAs-producing bacteria may be considered a novel interventive approach for the management of CP.

As an essential amino acid, l-tryptophan is widely used in food, feed and medicine sectors. Nowadays, microbial l-tryptophan production suffers from low productivity and yield. Here we construct a chassis E. coli TRP3 producing 11.80 g/L l-tryptophan, which was generated by knocking out the l-tryptophan operon repressor protein (trpR) and the l-tryptophan attenuator (trpL), and introducing the feedback-resistant mutant aroG^fbr. On this basis, the l-tryptophan biosynthesis pathway was divided into three modules, including the central metabolic pathway module, the shikimic acid pathway to chorismate module and the chorismate to tryptophan module. Then we used promoter engineering approach to balance the three modules and obtained an engineered E. coli TRP9. After fed-batch cultures in a 5 L fermentor, tryptophan titer reached to 36.08 g/L, with a yield of 18.55%, which reached 81.7% of the maximum theoretical yield. The tryptophan producing strain with high yield laid a good foundation for large-scale production of tryptophan.
Escherichia coli/metabolism* ; Tryptophan ; Metabolic Engineering ; Bioreactors ; Metabolic Networks and Pathways