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Additional file 1. All original and unprocessed images of Western blots.

Doxorubicin (DOX) remains the most effective anticancer agent which is widely used in several adult and pediatric cancers, but its application is limited for its cardiotoxicity and hepatotoxicity. Hydrogen, as a selective antioxidant, is a promising potential therapeutic option for many diseases. In this study, we found that intraperitoneal injection of hydrogen-rich saline (H

CDKL5 disorder is a severe neurodevelopmental disorder caused by mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene. It predominantly affects females that typically present with severe epileptic encephalopathy, intellectual disability, autistic features and motor dysfunction. Currently, there is no therapy apart from anti-epileptic drugs providing poor seizure management. To develop a gene replacement therapy, we initially characterised the CDKL5 isoforms expressed in human brain, neuronal cell lines, primary astrocytes and hESC-derived cortical interneurons, in which hCDKL5_1 and to a lesser extent hCDKL5_2 isoforms were found ubiquitously expressed. We cloned their coding regions downstream of the CBh promoter into ssAAV2 vector genome and produced high-titre rAAV vectors: AAV9, AAV-DJ for in vitro and AAV-PHP.B for in vivo applications. AAV-DJ vectors efficiently transduced CDKL5-mutant iPSC-derived neural progenitors, which were subsequently differentiated into neurons. hCDKL5_1 expression in CDKL5-mutant neurons led to an increased density of synaptic puncta, whilst hCDKL5_2 ameliorated the calcium signalling defect, implying distinct functions of these isoforms in neurons. Intrajugular delivery of AAV-PHP.B-GFP vectors in WT mice transduced neurons and astrocytes throughout the brain more efficiently than AAV9, and transduced spinal cord, DRGs, kidney and retina. Cdkl5 KO mice exhibited deficits in motor and cognitive behaviours compared to WT. Cdkl5 KO mice treated with AAV-PHP.B-hCDKL5_1 via intrajugular injection at age 28-30 days exhibited significant behavioural improvements compared to GFP-treated controls. Brain expression of hCDKL5_1 was more abundant in the hindbrain than the forebrain and midbrain. At the cellular level, transgene expression was sporadic in the brain and most prominent in CA1 hippocampal neurons and cerebellar Purkinje cells. Correction of PSD95 cerebellar misexpression, a major fine cerebellar structural abnormality in Cdkl5 KO mice, was found in regions of high transgene expression within cerebellum. This study provides the first evidence that AAV-mediated gene therapy can be effective in treating CDKL5 disorder. Open Access

A new zeolite molecular sieve modified with chitosan was prepared and employed for the adsorption of ammonia and nitrate from underground water. Batch of experiments were performed to investigate the effects of various conditions on chitosan/ zeolite molecular sieve(CTS-Z) preparation, including the concentration of acetic acid, the concentration of chitosan, the stirring time and the stirring temperature. Characteristic of CTS-Z adsorption particles were analyzed by Scanning electron microscopy (SEM), Specific surface area (BET), Fourier infrared spectroscopy (FTIR) and X ray photoelectron spectroscopy (XPS). Experimental results showed that the optimum preparation processes for the chitosan / zeolite molecular sieve were as follows: the concentration of acetic acid was 4%, the concentration of chitosan was 7g / L, the stirring time was 10h and the stirring temperature was 30°C. And adsorption capacity of adsorption particles for ammonia and nitrate were 0.636 mg/g and 1.952 mg/g, and the removal rates were 81.60 % and 40.28 % respectively. The surface morphology of the particles showed more protrusions and micropores, the specific surface area of the particle was 391.516 m2/g. FTIR analysis showed that CH3- and NH2- have been loaded into the basic framework of zeolite molecular sieve and XPS analysis showed that the element O 1s played an important role in the attachment of chitosan to zeolite molecular sieves. Therefore, the results of the research can provide a theoretical basis for the upgrading of the water purification plants in the cold region of northern China.

Cyclin-dependent-like kinase 5 (CDKL5) gene mutations lead to an X-linked disorder that is characterized by infantile epileptic encephalopathy, developmental delay, and hypotonia. However, we found that a substantial percentage of these patients also report a previously unrecognized anamnestic deficiency in pain perception. Consistent with a role in nociception, we found that CDKL5 is expressed selectively in nociceptive dorsal root ganglia (DRG) neurons in mice and in induced pluripotent stem cell (iPS)-derived human nociceptors. CDKL5-deficient mice display defective epidermal innervation, and conditional deletion of CDKL5 in DRG sensory neurons impairs nociception, phenocopying CDKL5 deficiency disorder in patients. Mechanistically, CDKL5 interacts with calcium/calmodulin-dependent protein kinase II α (CaMKIIα) to control outgrowth and transient receptor potential cation channel subfamily V member 1 (TRPV1)-dependent signaling, which are disrupted in both CDKL5 mutant murine DRG and human iPS-derived nociceptors. Together, these findings unveil a previously unrecognized role for CDKL5 in nociception, proposing an original regulatory mechanism for pain perception with implications for future therapeutics in CDKL5 deficiency disorder.