Recommendation of SLM Process Parameters Based on Analytic Hierarchy Process and Weighted Particle Swarm Optimization for High-Temperature Alloys.

Selective laser melting (SLM) of high-temperature alloys involves intricate interdependencies among key process parameters, such as laser power and scanning speed, affecting properties such as density and tensile strength. However, relying solely on experiential knowledge for process parameter design often hampers the precise attainment of target requirements. To address this challenge, we propose an innovative approach that integrates the analytic hierarchy process (AHP) and weighted particle swarm optimization (WPSO) to recommend SLM process parameters for high-temperature alloy fabrication. Our proposed AHP-WPSO model consists of three main steps. First, a comprehensive historical database is established, capturing the process parameters and performance metrics of high-temperature alloy SLM parts. Utilizing an AHP framework, we compute the performance similarity between target and historical cases, applying rational thresholds to identify analogous cases. When suitable analogs are elusive, the model seamlessly transitions to the second step. Here, the WPSO model optimizes and recommends process parameters according to target specifications. Lastly, our experimental validation of the GH4169 high-temperature alloy through SLM experiments corroborates the effectiveness of our AHP-WPSO model in making process parameter recommendations. The outcomes underscore the model's high accuracy, attaining a recommendation precision of 99.81% and 96.32% when historical analogs are present and absent, respectively. This innovative approach offers a robust and reliable solution to the challenges posed in SLM process parameter optimization for high-temperature alloy applications.

CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus Glarea lozoyensis and Its Application in Manipulating gloF.

Glarea lozoyensis is an important industrial fungus that produces the pneumocandin B0, which is used for the synthesis of antifungal drug caspofungin. However, because of the limitations and complications of traditional genetic tools, G. lozoyensis strain engineering has been hindered. In this study, we established an efficient CRISPR/Cas9-based gene editing tool in G. lozoyensis SIPI1208. With this method, gene mutagenesis efficiency in the target locus can be up to 80%, which enables the rapid gene knockout. According to the reports, GloF and Ap-HtyE, proline hydroxylases involved in pneumocandin and Echinocandin B biosynthesis, respectively, can catalyze the proline to generate different ratios of trans-3-hydroxy-l-proline to trans-4-hydroxy-l-proline. Heterologous expression of Ap-HtyE in G. lozoyensis decreased the ratio of pneumocandin C0 to (pneumocandin B0 + pneumocandin C0) from 33.5% to 11% without the addition of proline to the fermentation medium. Furthermore, the gloF was replaced by ap-htyE to study the production of pneumocandin C0. However, the gene replacement has been hampered by traditional gene tools since gloF and gloG, two contiguous genes indispensable in the biosynthesis of pneumocandins, are cotranscribed into one mRNA. With the CRISPR/Cas9 strategy, ap-htyE was knocked in and successfully replaced gloF, and results showed that the knock-in strain retained the ability to produce pneumocandin B0, but the production of pneumocandin C0 was abolished. Thus, this strain displayed a competitive advantage in the industrial production of pneumocandin B0. In summary, this study showed that the CRISPR/Cas9-based gene editing tool is efficient for manipulating genes in G. lozoyensis.

Danzhi Jiangtang Capsule Mediates NIT-1 Insulinoma Cell Proliferation and Apoptosis by GLP-1/Akt Signaling Pathway.

OBJECTIVE: This study aimed to investigate the effects of Danzhi Jiangtang Capsule (DJC) on the proliferation and apoptosis functions of NIT-1 pancreatic ß-cells exposed to high-glucose load through GLP-1 activated Akt/ FoxO1 signaling pathway. METHODS: Cellular apoptosis of NIT-1 pancreatic ß-cells was induced by culturing in medium with 33.3mmol/L high glucose (HG). Then low-dose DJC (HG +LD), high-dose DJC (HG +HD), high-dose DJC+ GLP-1 inhibition (HG +HD +GI), and high-dose DJC+AKT inhibition (HG +HD+AI) were added, respectively. Cellular proliferation was accessed by cell counting kit (CCK-8) and cellular apoptosis was measured by Annexin V-FITC/PI staining. The protein levels of phosphorylated phosphatidylinositol-3-kinase (p-PI3K), phosphorylated AKT (p-AKT), phosphorylated Forkhead box protein O1 (p-FoxO1), and cleaved caspase-3 were detected by Western blotting. The mRNA expression of pancreatic duodenal homeobox-1 (PDX-1), CyclinD1, Bcl-2, and insulin was tested by Q-PCR. RESULTS: Comparing to HG group, (HG+HD) group showed a significantly increased cellular proliferation. The apoptosis of NIT-1 cells also was obviously reduced, with downregulated cleaved caspase-3 protein level and upregulated PDX-1, CyclinD1, and Bcl-2 mRNA levels (P<0.05). Additionally, (HG+HD) group manifested increased insulin mRNA expression; the protein levels of p-PI3K and p-AKT were markedly increased and p-FoxO1 was decreased. All of the above therapeutic effects by DJC intervention had been reversed by GLP-1 inhibition in (HG+HD+GI) group or AKT inhibition in (HG+HD+AI) group. CONCLUSION: DJC was able to attenuate the toxicity of high-glucose load in NIT-1 pancreatic ß-cells, ascribed to the improvement of cellular proliferation and apoptosis by GLP-1/Akt signaling pathway. This study could supply a new mechanism of DJC effects on type 2 diabetes mellitus (T2DM) treatment.

Novel Polysaccharide H-1-2 from Pseudostellaria Heterophylla Alleviates Type 2 Diabetes Mellitus.

BACKGROUND/AIMS: To investigate the potential therapeutic effect of novel polysaccharide H-1-2 from pseudostellaria heterophylla against type 2 Diabetes Mellitus (T2DM) and elucidate the underling molecular mechanisms. METHODS: Relative expression of HIF1α and Sirt1 in T2DM patients was determined via real-time PCR. The direct binding of HIF1α on Sirt1 promoter was validated by ChIP assay. The inhibitory regulation of Sirt1 by HIF1α was analyzed using luciferase reporter assay. The endogenous protein of HIF1α and Sirt1 in response to H-1-2 treatment was quantified by western blotting. The blood glucose, secreted insulin and serous lipid profiles were measured with ELISA kits. RESULTS: We consolidated that HIF1α and Sirt1 was dysregulated in T2DM patients and subjected to H-1-2 modulation. H-1-2 significantly inhibited hypoxia and up-regulated Sirt1 expression in EndoC-ßH1 cells. Accordingly, H-1-2 enhanced glucose-stimulation insulin secretion and improved blood glucose and lipid profiles in T2DM cells, and elevated the glucose and insulin tolerance simultaneously. Furthermore, we demonstrated that H-1-2 alleviated T2DM via inhibition of hypoxia and up-regulation of Sirt1 in isolated pancreatic ß-cells from T2DM rats. CONCLUSION: Our data unambiguously demonstrated H-1-2 administration alleviated T2DM by enhancing Sirt1 expression through inhibition of hypoxia.

Enhancement of A82846B yield and proportion by overexpressing the halogenase gene in Amycolatopsis orientalis SIPI18099.

The glycopeptide antibiotic A82846B (chloroeremomycin) produced by Amycolatopsis orientalis is the precursor of the semi-synthetic antibiotic oritavancin. However, during the industrial production of A82846B, two major impurities, A82846A (63.6%) and A82846C (12%) which are structurally similar to A82846B (24.4%), are also produced. In this study, to improve the ratio of A82846B to A and C, the genes encoding halogenase in A82846B and vancomycin synthesis were integrated into A. orientalis SIPI18099 to test their halogenation ability, respectively. The results indicated that chal from the A82846B biosynthesis pathway was more efficient in reducing A and C factors. Moreover, by increasing the chal copy number, the proportion of A and C were gradually reduced while the titer and proportion of A82846B were improved. In a scaled-up industrial process, the proportion of A and C were decreased to 11.6% and 0.2% in the recombinant strain A.orientalis chal-3 with three gene copies of chal and the titers of A82846B (2.2 g/L) has increased by 2.8-folds compared to 780 mg/L produced by the parental strain, suggesting that the recombinant strain was suitable for the industrial production of A82846B with lower impurities.

[The effect of Salvianolic Acid B on the Apoptosis of HUVECs Induced by Intermittent High Glucose].

Objective: To investigate the effect of Salvianolic acid B (Sal B) on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by intermittent high glucose and to explore the possible mechanisms. Methods: HUVECs were preincubated with Sal B for 24 h, followed by incubation with intermittent high glucose (IHG, 5.5 mmol/L 12 h, 33.3 mmol/L 12 h) for 72 h. The viability of the HUVECs was determined by MTT assay, and the cells apoptosis was measured flow cytometry, respectively. The levels of nitric oxide (NO), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and Caspase-3 activity were determined by colorimetric method. Intracellular ROS was evaluated by fluorescent microscopy. The protein levels of NOX4, p-eNOS, BAX, and BCL-2 were determined by Western-blot. Results: Pretreatment with Sal B significantly ameliorated IHG-induced cells injury as was manifested by increased cell viability, up-regulated eNOS activation, and promoted the release of NO in HUVECs (P < 0.05 or P < 0.01). Sal B evidently suppressed IHG-induced cell apoptosis, down-regulated the expression of BAX protein and up-regulated the expression of BCL-2 protein. The activity of Capase-3 was also significantly reduced. Pre-incubation with Sal B led to a significant enhancement of antioxidant capacity and a reduction of NOX4 protein expression, accompanied by a remarkable decrease of intracellular ROS and MDA content (P < 0.05 or P < 0.01). Conclusion: Sal B is capable of suppressing IHG-induced injury and apoptosis in HUVECs, which might be attributed to the attenuation of oxidative stress, regulation of BCL-2/BAX protein expression, and subsequent suppression of Caspase-3 activity.

[Effect of Danzhi Jiangtang Capsule on Myocardial Fibrosis in Diabetic Rats].

OBJECTIVE: To investigate the effect of Danzhi Jiangtang Capsule on myocardial fibrosis in diabetic rats with fluctuated blood glucose and the possible mechanisms implicated. METHODS: Following induction of diabetes with intraperitoneal injection of streptozotocin (STZ), rats were administered with insulin or glucose at different time during a day to induce blood glucose fluctuation. After treatment with Danzhi Jiangtang Capsule for six weeks, rats were sacrificed and the hearts were collected for the determination of cardiac mass index. Cardiac levels of angiotensin II (Ang II), type I and type III collagens and transforming growth factor-ß1 (TGF-ß1) were assayed by ELISA. Levels of Smad3 phosphorylation and protein expression of matrix metalloproteinase-2 ( MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) were determined by Western blot analysis. Total antioxidant capacity and malondialdehyde (MDA) content in cardiac tissues were measured colorimetrically. RESULTS: Treatment with Danzhi Jiangtang Capsule for six weeks significantly reduced cardiac mass index and cardiac levels of type I and type III collagens (P < 0.05 or P < 0.01). Levels of Ang II, TGF-ß1 and Smad3 phosphorylation in cardiac tissues were also decreased markedly (P < 0.05 or P < 0.01). Supplementation with Danzhi Jiangtang Capsule resulted in an evident up-regulation of MMP-2 protein and down-regulation of TIMP-2 protein expression in cardiac tissues (P < 0.05 or P < 0.01). In addition, Danzhi Jiangtang Capsule significantly enhanced total antioxidant capacity in diabetic rats, while cardiac content of MDA was decreased markedly( P < 0.05 or P < 0.01). CONCLUSION: Danzhi Jiangtang Capsule significantly ameliorated myocardial fibrosis in diabetic rats with fluctuated blood glucose, which might be derived from enhancement of antioxidant capacity, suppression of RAS and TGF-ß1/Smad3 signaling pathway and regulation of MMP-2/TIMP-2 protein expression.

[Medication rule for treatment of functional dyspepsia: an analysis of traditional Chinese medicine literature based on China National Knowledge Internet].

By retrieving the clinical research literature of treatment functional dyspepsia by traditional Chinese medicine (TCM) from January 2004 to December 2014 based on China National Knowledge Internet (CNKI), we would establish a TCM decoction database for treating functional dyspepsia in this study. One hundred and sixty-four literature were included, involving 159 prescriptions, 377 medicines, in a total of 1 990 herbs. These herbs can be divided into 18 categories according to the effectiveness; and qi-regulating herbs, blood circulation herbs, and antipyretic herbs ranked top three ones according to the frequency of usage of the herbs, whose medicine usage frequency accounted for 51.81%. Usage frequency of 16 herbs was over 30, and Atractylodes, Radix, Poriaranked top three according to the usage frequency. Medicinal properties were divided into 9 kinds according to the frequency statistics, and the top three were warm, flat, and cold. Taste frequency statistics were classifiedinto 9 kinds, and the top three were acrid, sweet, and bitter. In frequency statistics of the meridian tropism of herbs, it was classifiedinto 11 kinds, and the top three were spleen, stomach, lung. The analysis can provide a reference for treatment and study of TCM of functional dyspepsia.

[Effect of danzhi jiangtang capsule combined exercise on the protein expression of NADPH oxidase p22phox in pancreatic tissues of diabetic rats].

OBJECTIVE: To explore effects of exercise combined Danzhi Jiangtang Capsule (DJC) on the protein expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase p22phox in pancreatic tissues of diabetic rats. METHODS: Sixty male Wistar rats were injected with low dose of streptozotocin and fed with high fat diet to establish a diabetic rat model. The levels of p22phox and 8-hydroxy-2-de-oxyguanosine (8-OHdG) protein in pancreatic tissues were detected by immunohistochemical method, and the level of p22phox protein was also detected by Western blot in the normal group, the model group, the excise group, the DJC group, and the DJC +excise group, respectively. RESULTS: The expression levels of p22phox and 8-OHdG protein in pancreatic tissues were significantly higher in the model group than in the normal group (P <0.01). p22phox and 8-OHdG were mainly expressed in the cytoplasm of pancreatic cells. After administration of exercise or DJC, the expression lev- els of p22phox or 8-OHdG protein in pancreatic tissues decreased significantly (P <0. 01). Exercise combined DJC had synergistic effects in decreasing expressions of p22phox and 8-OHdG (P <0.05). CONCLUSION: Exercise, DJC, and exercise combined DJC could protect islet beta cells by decreasing the expression of NADPH oxidase in beta cells and reducing sources of oxidative stress.

[Effects of Chinese herbal medicine Danzhi Jiangtang Capsule and exercise on JNK signaling pathway in pancreatic tissues of diabetic rats].

OBJECTIVE: To explore the effects of exercise and Danzhi Jiangtang Capsule (DJC), a compound traditional herbal medicine, on the JNK signaling pathway in pancreatic tissues of diabetic rats and to investigate the possible mechanisms of exercise and DJC in treating diabetes. METHODS: Seventy-eight male Wistar rats were injected with low dose of streptozotocin and fed a high-fat diet to establish a diabetic model in rats. Then 60 diabetic rats were divided into diabetes group, exercise group, DJC group and exercise combined with DJC group. Another twelve rats were used as normal control. After eight months of treatment, the expression levels of phosphor-c-Jun N-terminal kinase (p-JNK), pancreatic and duodenal homeobox-1 (PDX-1), and insulin protein in pancreatic tissues from rats were detected by immunohistochemical method and Western blotting. RESULTS: In pancreatic tissues of diabetes group, the expression level of p-JNK protein was significantly higher than that in the normal group (P<0.01), and the expression levels of PDX-1 and insulin protein were significantly decreased (P<0.01). After administration of exercise and DJC, the expression level of p-JNK protein in pancreatic tissues of the diabetes group was decreased significantly, while the expression levels of PDX-1 and insulin protein were increased significantly (P<0.05 or P<0.01). CONCLUSION: Exercise and DJC effectively protect isletß-cell function in diabetic rats, which might be due to a decreased JNK signaling pathway.

Protective effect of gliclazide on diabetic peripheral neuropathy through Drp-1 mediated-oxidative stress and apoptosis.

OBJECTIVE: To investigate the protective effect of gliclazide and the role of dynamin-related protein l (Drp-1)-mediated oxidative stress and apoptosis in diabetic peripheral neuropathy (DPN). METHODS: Diabetic rats developed through intra-peritoneal injection of streptozotocin were randomly assigned to treatment group receiving gliclazide or non-treatment group without gliclazide treatment. Rats in control group received intra-peritoneal injection of vehicle and no gliclazide treatment. Eight weeks later, the nerve conduction velocity (NCV) of sciatic nerve was measured and the morphological alterations, the malondialdehyde (MDA) level and superoxide-dismutase (SOD) activity, the expressions of Drp-1, caspase-3, Bax and Bcl-2 in sciatic nerve were evaluated. RESULTS: When compared to rats in control group, rats in non-treatment group showed significantly decrease of NCV, obvious demyelinative alteration of sciatic nerve, increased expressions of Drp-1, caspase-3, Bax, Bcl-2 and MDA, and decreased SOD activity. Compared to rats in non-treatment group, rats in treatment group showed significantly increase of NCV, less demyelination of sciatic nerve, decreased expressions of Drp-1, caspase-3, Bax and MDA, and increased activity of SOD. The expression of Bcl-2 was not significantly different between treatment and non-treatment groups. CONCLUSION: Gliclazide showed protective effect on DPN through modulating Drp-1-mediated oxidative stress and apoptosis.

[Effects of danzhi jiangtang Capsule combined exercise on pancreatic oxidative stress and islet beta-cell function in diabetic rats].

OBJECTIVE: To explore the effects of Danzhi Jiangtang Capsule (DJC) and exercise on islet beta-cell function index (HOMA-% beta), blood glucose, and oxidative stress of diabetic rats. METHODS: A diabetic rat model was established using low dose streptozotocin and high fat forage in 60 male Wistar rats. The effects of exercise, DJC, and DJC combined exercise on the serum levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), total cholesterol (TC), as well as the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) in the pancreatic tissue were observed. The HOMA-%beta was also calculated. The main factors that affecting HOMA-%beta were explored using multi-factor regression analysis. RESULTS: Compared with model group, the levels of FBG, TG, TC, and pancreatic MDA were significantly reduced after intervention of exercise or DJC (P < 0.05, P < 0.01), while HOMA-% beta obviously increased (P < 0.01). The pancreatic GSH-Px activity significantly increased in the exercise group (P < 0.01). Exercise and DJC had synergistic effects on FBG, TG, HOMA-% beta, pancreatic SOD, and GSH-Px activities (P < 0.05). There was a negative and linear regression correlation between FBG and pancreatic SOD and GSHPx activities. HOMA-%beta was negatively correlated with FBG, TG, TC, and pancreatic MDA content, and positively correlated with SOD and GSH-Px activities. Besides, there was a linear regression correlation between HOMA-%beta and FBG. CONCLUSION: Exercise and DJC played synergistic effects, could improve the glucose and lipid metabolisms and enhance antioxidant activities, thus relieving the injury of pancreatic beta cells.