Effects of transparent online open procurement on prices, volumes, and costs of medicines: an interrupted time series study in Ningxia, China.

Objectives: To assess the effects of the transparent online open procurement arrangement on the prices, volumes, and costs of medicines in Ningxia, China. Methods: Data were extracted from the Ningxia pharmaceutical procurement platform, covering 16 months of purchase orders (December 2019 to March 2021) prior to the implementation of the transparent online open procurement policy and 20 months of purchase orders after the implementation of the policy (April 2021 to November 2022). Interrupted time series (ITS) analysis was performed to evaluate the effects of the transparent online open procurement policy on the prices, volumes, and total costs of the purchase orders. Results: After implementation of the transparent online open procurement policy, the average price of purchased medicines showed a declining trend by 0.012 Yuan per month, while the total volume of purchase orders declined at a rate by 1.741 million per month measured by the smallest formulation units and the total costs of the purchase orders decreased at a rate by 5.525 million Yuan per month. Conclusion: The transparent online open procurement policy resulted in reduced prices, lowered volumes, and lowered total costs of purchased orders of medicines.

Policy text analysis of antimicrobial resistance governance in China: A focus on national-level policies.

OBJECTIVE: To explore the structure and characteristics of China's national policies regarding antimicrobial resistance (AMR) governance. METHOD: This research constitutes a quantitative content analysis of AMR policies issued by the central government from 2004 to 2023. A systematic search identified 112 policy documents, which were analysed using a three-dimensional framework. This framework included policy objectives, policy instruments (The supply-focused instrument aims to promote rational and prudent antibiotic prescriptions. The demand-focused instrument affects consumer use of antibiotics, either directly or indirectly. The environment-focused instrument provides a favourable and systematic policy environment for the prudent use of antibiotics.) and policy evolvement. Sub-themes under the framework were identified through a deductive process, followed by descriptions of frequency distributions of the sub-themes and categories. RESULTS: The majority of policy documents originated from individual governmental departments, with only 8 (7.14%) being jointly issued. The National Health Commission (NHC) remained the predominant policy maker, issuing 56 (48.21%) policies. A clear preference emerged for utilizing environment-focused policy instruments (69.70%), compared with the demand-focused (18.45%) and supply-focused (11.85%) instruments. 'Optimizing the use of antimicrobial medicines' ranked on top of the policy objectives, with 185 (31.25%) citations extracted across 74 (30.58%) policy documents. In addition to increasing numbers of policies over the three stages (2004-2011, 2012-2015, 2016-2023) of development, the use of various instruments became more comprehensive and balanced in the third stage. CONCLUSIONS: AMR governance has become increasingly comprehensive in China, despite a deficit in inter-sectoral collaborations. A whole-of-government approach is required to maximize the value of various policy initiatives.

Metabolomics and its applications in assisted reproductive technology.

Metabolomics, an emerging omics technology developed in the post-gene age, is an important part of systems biology. It interprets the pathophysiological state of the subject by quantitatively describing the dynamic changes of metabolites through analytical methods, mainly mass spectrometry (MS) and nuclear magnetic resonance (NMR). Assisted reproductive technology (ART) is a method used to manipulate sperm, oocytes, and embryos to achieve conception. Recently, several studies have reported that metabolomics methods can be used to measure metabolites in ART samples; these metabolites can be used to evaluate the quality of gametes and embryos. This article reviews the progress of research on metabolomics and the application of this technology in the field of ART, thus providing a reference for research and development directions in the future.

Oxymatrine inhibits the pyroptosis in rat insulinoma cells by affecting nuclear factor kappa B and nuclear factor (erythroid-derived 2)-like 2 protein/heme oxygenase-1 pathways.

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1ß and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1ß, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

Voltage-gated potassium channels are involved in oxymatrine-regulated islet function in rat islet ß cells and INS-1 cells.

OBJECTIVES: Oxymatrine can regulate glucose metabolism. But the underlying mechanisms remain unclear. We investigated the relationship of oxymatrine and voltage-gated potassium (Kv) channel in rat islet ß cells and INS-1 cells. MATERIALS AND METHODS: Insulin secretion and Kv channel currents were tested by radioimmunoassay and patch-clamp technique, respectively. The INS-1 cell viability was detected using cell counting kit-8 experiments. Flowcytometry analysis and western blot were employed for cell apoptosis and protein levels, respectively. INS-1 cell proliferation was assessed by the 5-Ethynyl-2'- deoxyuridine method. RESULTS: Oxymatrine potentiated insulin secretion at high glucose (P<0.01 vs 11.1 G, P<0.01 vs 16.7 G) and inhibited KV currents at 40 mV (45.73±15.34 pA/pF for oxymatrine, 73.80±19.23 pA/pF for control, P<0.05). After the INS-1 cells were treated with oxymatrine for 12 and 24 hr, KV2.1 channel protein was up-regulated (P<0.01 vs Control). At the same time, compared with the high glucose and high fat group, cell viability and proliferation ability were increased (P<0.01). The cell apoptotic rate was reduced, reaching 17.30%±1.00% at 12 hr and 10.35%±1.52% at 24 hr (P<0.01). These protective effects of oxymatrine were reversed by using Stromatoxin-1, a kv channel inhibitor. CONCLUSION: The results indicate that oxymatrine can stimulate insulin secretion and decrease kv channel currents in islet ß cells. Besides, oxymatrine also increases cell viability, proliferation, and reduces cell apoptosis in INS-1 cells. The effects of oxymatrine are related to kv channels. This finding provides new insight into the mechanisms of oxymatrine-regulated islet function.

The PLC/PKC/Ras/MEK/Kv channel pathway is involved in uncarboxylated osteocalcin-regulated insulin secretion in rats.

Uncarboxylated osteocalcin, a bone matrix protein, has been proposed to regulate glucose metabolism by increasing insulin secretion, improving insulin sensitivity and stimulating ß cell proliferation. Our previous study also indicated that uncarboxylated osteocalcin stimulates insulin secretion by inhibiting voltage-gated potassium (KV) channels. The goal of this study is to further investigate the underlying mechanisms for the regulation of Kv channels and insulin secretion by uncarboxylated osteocalcin. Insulin secretion and Kv channel currents were examined by radioimmunoassay and patch-clamp technique, respectively. Calcium imaging system was applied to measure intracellular Ca2+ concentration ([Ca2+]i). The protein levels were detected by western blot. The results showed that uncarboxylated osteocalcin potentiated insulin secretion, inhibited Kv channels and increased [Ca2+]i compared to control. These effects were suppressed by phospholipase-C (PLC)/protein kinase C (PKC)/Ras/MAPK-ERK kinase (MEK) signaling pathway, indicating that this signaling pathway plays an important role in uncarboxylated osteocalcin-regulated insulinotropic effect. In addition, the results also showed that adenylyl cyclase (AC) did not influence the effect of uncarboxylated osteocalcin on insulin secretion and Kv channels, suggesting that AC is not involved in uncarboxylated osteocalcin-stimulated insulin secretion. These findings provide new insight into the mechanism of uncarboxylated osteocalcin-regulated insulin secretion.

Geniposide acutely stimulates insulin secretion in pancreatic ß-cells by regulating GLP-1 receptor/cAMP signaling and ion channels.

Geniposide, an iridoid glycoside, has antidiabetic effects. The present study aimed to evaluate whether geniposide has direct effects on insulin secretion from rat pancreatic islets. The results demonstrated that geniposide potentiated insulin secretion via activating the glucagon-like-1 receptor (GLP-1R) as well as the adenylyl cyclase (AC)/cAMP signaling pathway. Inhibition of protein kinase A (PKA) suppressed the insulinotropic effect of geniposide. Geniposide also inhibited voltage-dependent potassium (Kv) channels, and this effect could be attenuated by inhibition of GLP-1R or PKA. Current-clamp recording showed that geniposide prolonged action potential duration. These results collectively imply that inhibition of Kv channels is linked to geniposide-potentiated insulin secretion by acting downstream of the GLP-1R/cAMP/PKA signaling pathway. Moreover, activation of Ca(2+) channels by geniposide was observed, indicating that the Ca(2+) channel is also an important player in the geniposide effects. Together, these findings provide new insight into the mechanism underlying geniposide-regulated insulin secretion.

Inhibition of voltage-gated potassium channels mediates uncarboxylated osteocalcin-regulated insulin secretion in rat pancreatic ß cells.

Insulin secretion from pancreatic ß cells is important to maintain glucose homeostasis and is regulated by electrical activities. Uncarboxylated osteocalcin, a bone-derived protein, has been reported to regulate glucose metabolism by increasing insulin secretion, stimulating ß cell proliferation and improving insulin sensitivity. But the underlying mechanisms of uncarboxylated osteocalcin-modulated insulin secretion remain unclear. In the present study, we investigated the relationship of uncarboxylated osteocalcin-regulated insulin secretion and voltage-gated potassium (KV) channels, voltage-gated calcium channels in rat ß cells. Insulin secretion was measured by radioimmunoassay. Channel currents and membrane action potentials were recorded using the conventional whole-cell patch-clamp technique. Calcium imaging system was used to analyze intracellular Ca(2+) concentration ([Ca(2+)]i). The data show that under 16.7mmol/l glucose conditions uncarboxylated osteocalcin alone increased insulin secretion and [Ca(2+)]i, but with no such effects on insulin secretion and [Ca(2+)]i in the presence of a KV channel blocker, tetraethylammonium chloride. In the patch-clamp experiments, uncarboxylated osteocalcin lengthened action potential duration and significantly inhibited KV currents, but had no influence on the characteristics of voltage-gated calcium channels. These results indicate that KV channels are involved in uncarboxylated osteocalcin-regulated insulin secretion in rat pancreatic ß cells. By inhibiting KV channels, uncarboxylated osteocalcin prolongs action potential duration, increases intracellular Ca(2+) concentration and finally promotes insulin secretion. This finding provides new insight into the mechanisms of osteocalcin-modulated insulin secretion.

PI3K is involved in P2Y receptor-regulated cAMP /Epac/Kv channel signaling pathway in pancreatic ß cells.

P2Y receptors (P2YR) are a family of purinergic G protein-coupled receptors, which could be stimulated by extracellular nucleotides. In pancreatic ß cells, activation of P2YR has long been shown to stimulate insulin secretion in a glucose-dependent manner. Previously, we reported that P2YR-modulated insulin secretion is mediated by a cAMP/Epac/Kv channel pathway. However, the interaction between Epac and the Kv channel in P2YR-modulated insulin secretion remains unclear. In this study, we used patch-clamp technique and insulin secretion assay to investigate the potential molecules that may link Epac to Kv channel inhibition induced by P2YR activation. We identified that phosphatidylinositide 3-kinase, which mediates P2YR-regulated insulin secretion, is a critical mediator between Epac and the Kv channel.

P2Y purinergic receptor-regulated insulin secretion is mediated by a cAMP/Epac/Kv channel pathway.

Enhancement of insulin secretion is a major therapeutic approach for type 2 diabetes (T2D). Activation of P2Y purinergic receptor (P2YR) causes potentiation of insulin secretion in a glucose-dependent manner, making it a promising therapeutic target for T2D. Here we show that activation of P2YR to potentiate insulin secretion is mediated by adenylyl cyclase/cyclic AMP (cAMP) and the downstream effector, exchange protein directly activated by cAMP (Epac), leading to inhibition of voltage-dependent potassium (Kv) channels. P2YR-mediated Kv channel inhibition results in prolongation of action potential duration, and in turn elevates intracellular Ca(2+) level and insulin secretion. Taken together, the data indicate that cAMP/Epac/Kv channel pathway mediates P2YR-regulated insulin secretion, which may have important therapeutic implications for T2D.

The adenylyl cyclase inhibitor MDL-12,330A potentiates insulin secretion via blockade of voltage-dependent K(+) channels in pancreatic beta cells.

OBJECTIVE: Adenylyl cyclases (ACs) play important role in regulating pancreatic beta cell growth, survival and secretion through the synthesis of cyclic AMP (cAMP). MDL-12,330A and SQ 22536 are two AC inhibitors used widely to establish the role of ACs. The goal of this study was to examine the effects of MDL-12,330A and SQ 22536 on insulin secretion and underlying mechanisms. METHODS: Patch-clamp recording, Ca(2+) fluorescence imaging and radioimmunoassay were used to measure outward K(+) currents, action potentials (APs), intracellular Ca(2+) ([Ca(2+)]i) and insulin secretion from rat pancreatic beta cells. RESULTS: MDL-12,330A (10 µmol/l) potentiated insulin secretion to 1.7 times of control in the presence of 8.3 mmol/l glucose, while SQ 22536 did not show significant effect on insulin secretion. MDL-12,330A prolonged AP durations (APDs) by inhibiting voltage-dependent K(+) (KV) channels, leading to an increase in [Ca(2+)]i levels. It appeared that these effects induced by MDL-12,330A did not result from AC inhibition, since SQ 22536 did not show such effects. Furthermore, inhibition of the downstream effectors of AC/cAMP signaling by PKA inhibitor H89 and Epac inhibitor ESI-09, did not affect KV channels and insulin secretion. CONCLUSION: The putative AC inhibitor MDL-12,330A enhances [Ca(2+)]i and insulin secretion via inhibition of KV channels rather than AC antagonism in beta cells, suggesting that the non-specific effects is needed to be considered for the right interpretation of the experimental results using this agent in the analyses of the role of AC in cell function.