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In order to understand the genetic variation and whole genome characteristics of goose as-trovirus(GoAstV)in Jiangxi Province,a strain of GoAstV was successfully isolated from a typical gosling gout case sample and the whole genome sequencing and genetic characteristics of the isola-ted strain were analyzed.The results showed that the JXNC1 strain could be stably passaged on LMH cells and could cause mild cytopathic effects in LMH cells.Sequencing and analysis showed that the full-length genome of the strain was 7 173 bp,and its genetic relationship was the closest to the GZ2301(PP966939)reference strain,belonging to the GoAstV-Ⅱ genotype.The complete genome of JXNC1 strain shared 98.1％-98.8％nucleotide similarity with 22 GoAstV-Ⅱ reference strains,and the amino acid similarity of ORF2 gene was 97.6％-99.6％.At the same time,the analysis results showed that the mutation of the strain mainly occurred in the ORF2 gene,and there were 13 amino acid site mutations,of which T630I was a unique mutation.Animal regression experiments showed that the inoculation of JXNC1 strain could cause urate deposition in the or-gans of goslings,congestion and dilatation of hepatic sinusoids,and small focal necrosis of some hepatocytes.Renal tissue tubular dilatation,renal interstitial connective tissue hyperplasia.The re-sults of this study laid a foundation for accurate prevention and control of the disease.

In order to understand the genetic variation and whole genome characteristics of goose as-trovirus(GoAstV)in Jiangxi Province,a strain of GoAstV was successfully isolated from a typical gosling gout case sample and the whole genome sequencing and genetic characteristics of the isola-ted strain were analyzed.The results showed that the JXNC1 strain could be stably passaged on LMH cells and could cause mild cytopathic effects in LMH cells.Sequencing and analysis showed that the full-length genome of the strain was 7 173 bp,and its genetic relationship was the closest to the GZ2301(PP966939)reference strain,belonging to the GoAstV-Ⅱ genotype.The complete genome of JXNC1 strain shared 98.1％-98.8％nucleotide similarity with 22 GoAstV-Ⅱ reference strains,and the amino acid similarity of ORF2 gene was 97.6％-99.6％.At the same time,the analysis results showed that the mutation of the strain mainly occurred in the ORF2 gene,and there were 13 amino acid site mutations,of which T630I was a unique mutation.Animal regression experiments showed that the inoculation of JXNC1 strain could cause urate deposition in the or-gans of goslings,congestion and dilatation of hepatic sinusoids,and small focal necrosis of some hepatocytes.Renal tissue tubular dilatation,renal interstitial connective tissue hyperplasia.The re-sults of this study laid a foundation for accurate prevention and control of the disease.

Parabacteroides distasonis is a gram-negative bacterium initially isolated from a clinical specimen in the 1930s. The strain was re-classified to form the new genus Parabacteroides in 2006. P. distasonis can regulate intestinal barrier function and plays a key role in immune response and metabolic regulation of bodies. Traditional Chinese medicine(TCM) is closely related to the intestinal microbiota. Polysaccharides, saponins, and other ingredients of TCM can treat diseases by interacting with P. distasonis, but the specific mechanisms underlying these processes are still unclear, requiring further exploration. This study reviewed the roles and related mechanisms of P. distasonis in inflammatory-immune diseases, metabolic diseases, cardiovascular disease, neuropsychiatric diseases, cancer, and other diseases and summarized the relevant research results of TCM to prevent and treat diseases by regulating P. distasonis. This study provides a reference for subsequent exploration of P. distasonis and research on the interaction between TCM and intestinal microbiota.
Humans ; Gastrointestinal Microbiome/drug effects* ; Medicine, Chinese Traditional ; Animals ; Bacteroidetes ; Drugs, Chinese Herbal/pharmacology*

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGAD^{S24R/D88R/Y309K} was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and Lpgad^{S24R/D88R/Y309K} genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD₆₀₀) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.
Glutamate Decarboxylase/genetics* ; Lactobacillus plantarum/genetics* ; Catalysis ; gamma-Aminobutyric Acid ; Hydrogen-Ion Concentration ; Glutamic Acid

OBJECTIVE: To investigate the value of coronary computed tomographic angiography (CCTA)-based fractional flow reserve (CT-FFR) and plaque quantitative analysis in predicting adverse outcomes in patients with non-obstructive coronary heart disease (CAD). METHODS: Clinical data of patients with non-obstructive CAD who underwent CCTA at the Affiliated Hospital of Jiangnan University from March 2014 to March 2018 were retrospectively analyzed and followed up, and the occurrence of major adverse cardiovascular event (MACE) was recorded. The patients were divided into MACE and non-MACE groups according to the occurrence of MACE. The clinical data, CCTA plaque characteristics including plaque length, stenosis degree, minimum lumen area, total plaque volume, non-calcified plaque volume, calcified plaque volume, plaque burden (PB) and remodelling index (RI), and CT-FFR were compared between the two groups. Multivaritate Cox proportional risk model was used to evaluate the relationship between clinical factors, CCTA parameters and MACE. The receiver operator characteristic curve (ROC curve) was used to assess the predictive power of outcome prediction model based on different CCTA parameters. RESULTS: Finally 217 patients were included, of which 43 (19.8%) had MACE and 174 (80.2%) did not. The median follow-up interval was 24 (16, 30) months. The CCTA showed that patients in the MACE group had more severe stenosis than that in the non-MACE group [(44.3±3.8)% vs. (39.5±2.5)%], larger total plaque volume and non-calcified plaque volume [total plaque volume (mm³): 275.1 (197.1, 376.9), non-calcified plaque volume (mm³): 161.5 (114.5, 307.8) vs. 117.9 (77.7, 185.5)], PB and RI were larger [PB: 50.2% (42.1%, 54.8%) vs. 45.1% (38.2%, 51.7%), RI: 1.19 (0.93, 1.29) vs. 1.03 (0.90, 1.22)], CT-FFR value was lower [0.85 (0.80, 0.88) vs. 0.92 (0.87, 0.97)], and the differences were statistically significant (all P < 0.05). Cox regression analysis showed that non-calcified plaques volume [hazard ratio (HR) = 1.005. 95% confidence interval (95%CI) was 1.025-4.866], PB ≥ 50% (HR = 3.146, 95%CI was 1.443-6.906), RI ≥ 1.10 (HR = 2.223, 95%CI was 1.002-1.009) and CT-FFR ≤ 0.87 (HR = 2.615, 95%CI was 1.016-6.732) were independent predictors of MACE (all P < 0.05). The model based on CCTA stenosis degree+CT-FFR+quantitative plaque characteristics (including non-calcified plaque volume, RI, PB) [area under the ROC curve (AUC) = 0.91, 95%CI was 0.87-0.95] had significantly better predictive efficacy for adverse outcomes than the model based on CCTA stenosis degree (AUC = 0.63, 95%CI was 0.54-0.71) and the model based on CCTA stenosis degree+CT-FFR (AUC = 0.71, 95%CI was 0.63-0.79; both P < 0.01). CONCLUSIONS CT-FFR and plaque quantitative analysis based on CCTA are helpful in predicting adverse outcomes in patients with non-obstructive CAD. Non-calcified plaque volume, RI, PB and CT-FFR are important predictors of MACE. Compared with the prediction model based on stenosis degree and CT-FFR, the combined plaque quantitative index can significantly improve the prediction efficiency of adverse outcomes in patients with non-obstructive CAD.
Humans ; Fractional Flow Reserve, Myocardial ; Coronary Angiography/methods* ; Constriction, Pathologic ; Retrospective Studies ; ROC Curve ; Predictive Value of Tests ; Plaque, Atherosclerotic/diagnostic imaging* ; Coronary Stenosis/diagnostic imaging* ; Tomography, X-Ray Computed ; Coronary Artery Disease/diagnostic imaging*

Objective:To explore the effect of sciatic nerve derived exosomes(SN-EXO) on axon regeneration and functional recovery after peripheral nerve injury(PNI).Methods:From March 2021 to October 2022, the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University studied the effect of SN-EXO on the proliferation of Schwann cells(SCs) through EdU cell proliferation experiment. Twenty-one healthy male SD rats were randomly divided into 3 groups of sham operation, peripheral nerve injury(PNI) and SN-EXO treatment, with 7 rats in each group. The right sciatic nerves of rat models in sham group were exposed without injury. In the rat in PNI group and SN-EXO treatment group, PBS and SN-EXO were injected under the epineurium of right sciatic nerves following sciatic nerve crush. Sciatic nerve function index(SFI) was performed at 28 days after operation, and then sacrificed. Right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerves were evaluated by HE staining. Regeneration efficiency of neurofilaments and SCs were obserred by NF200 and S100β double staining of sciatic nerve. The data obtained were statistically analyzed, and P<0.05 was statistically significant. Results:It was found that SN-EXO can significantly enhance the proliferation ability of SCs, with statistically significant difference( P<0.05). SFI in SN-EXO treatment group and PNI group were(-27.65±4.36) and(-57.33±7.49), respectively, and the difference was statistically significant( P<0.05). Axons in SN-EXO treatment group were arranged more closely and orderly than those in the PNI group at 28 days after operation, and there were less injury induced axon disintegration and vacuolation. Immunofluorescence assay indicated that NF200 and S100β fluorescence intensity in SN-EXO treatment group was significantly higher than that in the PNI group, and the difference was statistically significant( P<0.05). Conclusion:SN-EXO could enhance the proliferation of SCs to promote axon regeneration following peripheral nerve injury.

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

Limonene and its derivative perillic acid are widely used in food, cosmetics, health products, medicine and other industries as important bioactive natural products. However, inefficient plant extraction and high energy-consuming chemical synthesis hamper the industrial production of limonene and perillic acid. In this study, limonene synthase from Mentha spicata was expressed in Saccharomyces cerevisiae by peroxisome compartmentalization, and the yield of limonene was 0.038 mg/L. The genes involved in limonene synthesis, ERG10, ERG13, tHMGR, ERG12, ERG8, IDI1, MVD1, ERG20ww and tLS, were step-wise expressed via modular engineering to study their effects on limonene yield. The yield of limonene increased to 1.14 mg/L by increasing the precursor module. Using the plasmid with high copy number to express the above key genes, the yield of limonene significantly increased up to 86.74 mg/L, which was 4 337 times higher than that of the original strain. Using the limonene-producing strain as the starting strain, the production of perillic acid was successfully achieved by expressing cytochrome P450 enzyme gene from Salvia miltiorrhiza, and the yield reached 4.42 mg/L. The results may facilitate the construction of cell factory with high yield of monoterpene products by S. cerevisiae.
Saccharomyces cerevisiae/metabolism* ; Limonene/metabolism* ; Metabolic Engineering ; Monoterpenes/metabolism*

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.