BAPTA-AM is a standard tool agent of calcium chelating.After entering into cells,BAPTA-AM is metabolized by lipase and forms BAPTA.The latter combines with calcium ion rapidly and high selectivity.The effects of BAPTA-AM can effectively protect nerve cells against hypoxic/ischemic injury and protect hepatocytes against cytotoxins attack.BAPTA-AM is a hopeful candidate for treating stroke,liver failure and other clinical critical diseases.

The blood-brain barrier(BBB) is formed by the brain capillary endothelium and excludes from the brain approximately 100% of large-molecule neurotherapeutics and more than 98% of all small-molecule drugs.There are three routines to increase drug transport into brain,ie,neurosurgical,pharmaceutical chemical,and physiological ways and the later is most hopeful.The receptor-mediated transcytosis system on the brain capillary endothelium could be used as multifunctional vectors for drug active transport into brain.This provides the platform for CNS drug delivery programs.

Aim To investigate the transcytosis mechanism of chlorogenic acid(CGA)by using Caco-2 and MDCK(Madin Darby canine kidney) monolayers models.Method ① Caco-2 and MDCK cell models:Caco-2 cell(105 cells/cm2) and MDCK cell(5?104 cells/cm2) were inoculated in Millicell-CM culture plate inserts,and the TEER of cell monolayer were detected to make sure the models are available for experiments.② Permeating experiments: to measure the value of OD of CGA and calculate the cumulative amount.Result CGA could be Absorbed and secreted on two monolayer models.Verapamil could inhibit the secretion at lower concentration of CGA on MDCK monolayer model.P-pg could partly act on the secretion of CGA on Caco-2 and MDCK cell models.Conclusion CGA can secrete and Absorb at the same time across Caco-2 and MDCK cell monolayers,P-pg partly involving in the secretion of CGA.

Calpains are the family of Ca2+-activated cysteine pro-teases. Although calpains are considered to be cytoplasmic en-zymes, recent research has demonstrated that μ-calpain, m-cal-pain, calpain 10 and their endogenous inhibitor calpastatin are present in the mitochondria and play important roles both in caspase -dependent and-independent pathways in cell death phe-nomena. Calpains exert direct and indirect effects on the caspases
and regulators of apoptosis pathway such as Bcl-2, Bax, Bid, promoting the release of Cyt-C, AIF, then result in cellular ap-optosis. To allow pharmacological targeting of these enzymes, thorough knowledge of their patterns of activation and further in-teractions with already known apoptotic pathways is necessary.

AIM　To study the role of 5-HT in antinocieption produced by intrathecal indomethacin (Ind) in mice. METHOD　The antinocieption of indomethacin was investigated on the tail immersion test in mice；the contents of 5-HT, 5-HAA were assayed with fluorescent method. RESULT　Dose-dependent antinocieption was observed following intrathecal administration of Ind 1.8 mg*kg-1, the effect could be obliterated when the animals were pretreated with cyproheptadine. After intrathecal administration of Ind, the content of 5-HIAA in the spinal cord of mice was significantly increased, but PG had no effect. CONCLUSION　The result imply that intrathecal indomethacin could produce antinocieption; this effect is mediated by 5-HT. PG does not participate in the action of 5-HT.

Skeletal muscle injury is a common disease in clinical practice,and an in-depth understanding of its repair mechanisms is crucial for the development of effective therapeutic strategies.This paper focuses on the key role of TGF-β in skeletal muscle injury repair,introduces the diversity of its family members and signaling pathways,explores the expression and regulation part of TGF-β after skeletal muscle injury,analyzes its early expression dynamics and regulatory factors,and thoroughly investigates the effects of TGF-β on skeletal muscle repair,revealing its inflammatory regulation,cellular activation and proliferation as well as fibrosis.Key role.Special attention was paid to its mechanism of action in muscle regeneration and its regulatory mechanism at the cellular level.In addition,the potential clinical applications of TGF-β in the repair of skeletal muscle injury were discussed,and the development and application of it as a therapeutic target and modulator were explored.However,controversies and shortcomings still exist in the current study,such as the dual roles of TGF-β and the impact of individual differences on treatment.Future research directions should include digging deeper into the details of signaling pathways and biomarker discovery.By overcoming these challenges,the potential clinical application of TGF-β in skeletal muscle injury repair is expected to usher in new breakthroughs and provide patients with more individualized and effective treatment strategies.

Objective To investigate the neuroprotective effect of methylene blue on diabetic retinopathy in rats. Methods Thirty SD rats were randomly divided into blank, control and experimental groups. The control and experimental groups were induced with diabetes by streptozotocin (STZ) intraperitoneal injection. After 6 weeks of successful modeling, the experimental group received intravitreal injection of methylene blue at a dose of [0.2 mg/(kg.d)], while the control group received an equal amount of dimethyl sulfoxide (DMSO) intravitreal injection, both continuously injected for 7 days. ELISA was used to detect the levels of retinal superoxide dismutase (SOD), 8-iso-prostaglandin F2alpha (iPF2α) and interleukin-1β (IL-1β) in rats. Western blot analysis was used to detect the expression of retinal extracellular signal-regulated kinase 1/2 phosphorylation (p-ERK1/2) and phosphorylated protein kinase B (p-AKT), and PAS staining was used to detect retinal morphological changes. Results Compared with the blank group rats, the retinal SOD activity in the control and experimental group rats was significantly reduced. iPF2α, IL-1β and p-ERK1/2 level increased, while p-AKT level decreased. Compared with the control group, the SOD activity of the experimental group rats increased. iPF2α and IL-1β level went down, while p-ERK1/2 and p-AKT level went up significantly. The overall thickness of the retinal layer and the number of retinal ganglion cells were significantly reduced. Conclusion Methylene blue improves diabetic retinopathy in rats by reducing retinal oxidative stress and enhancing ERK1/2 and AKT phosphorylation.
Rats ; Animals ; Diabetic Retinopathy/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Mitogen-Activated Protein Kinase 3/metabolism* ; Interleukin-1beta/metabolism* ; Methylene Blue/pharmacology* ; Phosphorylation ; Rats, Sprague-Dawley ; MAP Kinase Signaling System ; Diabetes Mellitus, Experimental/drug therapy* ; Superoxide Dismutase/metabolism*