HSPA8 inhibitors augment cancer chemotherapeutic effectiveness via potentiating necroptosis.

Our recent work has uncovered a novel function of HSPA8 as an amyloidase, capable of dismantling the RHIM-containing protein fibrils to suppress necroptosis. However, the impact of HSPA8 inhibitors on cancer regression via necroptosis remains unexplored. In this study, we conducted a comprehensive investigation to assess the potential of HSPA8 inhibitors in enhancing necroptosis both in vitro and in vivo. Our findings indicate that pharmacologic inhibition of HSPA8, achieved either through VER (VER-155008) targeting the NBD domain or PES (pifithrin-µ) targeting the SBD domain of HSPA8, significantly potentiates necroptosis induced by diverse treatments in cellular assays. These inhibitors effectively disrupt the binding of HSPA8 to the RHIM protein, impeding its regulatory function on RHIM amyloid formation. Importantly, HSPA8 inhibitors significantly enhanced cancer cell sensitivity to microtubule-targeting agents (MTAs) in vitro, while reversing chemoresistance and facilitating tumor regression by augmenting necroptosis in vivo. Our findings suggest a promising therapeutic approach to cancer through necroptosis modulation via HSPA8 targeting, particularly in combination with MTA drugs for enhanced treatment efficacy.

Long-Acting ß2 Adrenergic Receptor Agonist Ameliorates Imiquimod-Induced Psoriasis-Like Skin Lesion by Regulating Keratinocyte Proliferation and Apoptosis.

Psoriasis is a chronic inflammatory disease that affects approximately 1%-5% of the population worldwide. Considering frequent relapse, adverse drug reactions, and large costs of treatment, it is urgent to identify new medications for psoriasis. Keratinocytes play an essential role during psoriasis development, and they express high levels of ß2-Adrenergic receptor (ß2-AR), which increases intracellular cAMP levels when activated. Increased level of cAMP is associated with the inhibition of epidermal cell proliferation. In the present study, we observed the effect of salmeterol, a long-acting ß2-AR agonist, on the proliferation and apoptosis of keratinocytes as well as imiquimod-induced psoriasis-like skin lesions in mice. As phosphodiesterase 4 (PDE4) inhibitors increases intracellular cAMP concentration by inhibiting its inactivation, we further explored the synergetic effect of a PDE4 inhibitor and salmeterol on psoriasis-like skin lesions in mice. Our results indicated that salmeterol effectively inhibited the proliferation of HaCaT cells induced by TNF-α and serum, and this effect was accompanied by significantly increased apoptosis and CREB phosphorylation, which were reversed by the PKA inhibitor, H89. Salmeterol ameliorated imiquimod-induced psoriasis-like skin lesions in mice, but salmeterol combined with a PDE4 inhibitor had no synergetic effect in improving skin lesions in mice. Of note, the synergistic effects of anti-proliferation and induction of apoptosis in HaCaT cells appeared by inhibiting ERK signaling. In summary, salmeterol, a long-acting ß2-AR agonist, alleviates the severity of psoriasis via inhibiting the proliferation and promoting apoptosis of keratinocytes, partially by activating the cAMP/PKA signaling pathway.

PDE4D binds and interacts with YAP to cooperatively promote HCC progression.

The role of cAMP in the development of hepatocellular carcinoma (HCC) is controversial and the biological function of cAMP-hydrolysing enzyme phosphodiesterase 4D (PDE4D) in HCC remains unclear. In this study, we observed markedly higher PDE4D expression in HCC patients with poor survival. PDE4D bound to yes-associated protein (YAP), and PDE4D expression positively correlated with YAP expression in HCC. Overexpression of PDE4D increased YAP dephosphorylation and activity and promoted HCC cell growth in vitro and in vivo, which was attenuated by the YAP inhibitor verteporfin. In contrast, silencing PDE4D reduced YAP expression and HCC cell growth. Notably, forced expression of YAP promoted PDE4D and YAP target gene expression and cell growth, which were abrogated by the PDE4D inhibitor roflumilast. Mechanistically, silencing of YAP caused PDE4D downregulation and HCC cell apoptosis via extracellular signal-regulated kinase (ERK) activation. Roflumilast activated cAMP-PKA signaling and induced cAMP-PKA-dependent YAP phosphorylation at serine 127, resulting in YAP degradation and suppression of HCC growth, which were reversed by the PKA inhibitor PKI. Additionally, transfection of the YAP-S127A mutant reversed roflumilast-mediated suppression of YAP and cell growth. Taken together, our findings indicate that PDE4D binds to and interacts with YAP to promote HCC progression. Targeting the PDE4D-YAP interaction with roflumilast may be an effective strategy for HCC treatment.

Analysis on Population Differences in Motivation of Healthy Subjects to Participate in Phase I Clinical Trials / 中国医学伦理学

【Objective:】 To understand the differences in motivation of different groups to participate in phase I clinical trials, and to provide suggestions for the recruitment and management of healthy subjects in research institutions, so as to improve the quality of clinical trials and protect the subjects’ rights and interests. 【Methods:】 Questionnaire survey was conducted among healthy subjects and potential community subjects in phase I clinical trials being carried out in a tertiary A hospital in Beijing from 2020 to 2021. SAS 9.2 was used for statistical analysis of data. 【 Results:】 There were many dimension differences between phase I ward subjects and community potential subjects in demographic and sociological characteristics, motivation to participate in the trials and influencing factors of motivation to participate in the experiment. Subjects in phase I ward paid more attention to the trial compensation, health care, social interaction and making friends, and the attitude of family or friends towards their participation in the trials. Community potential subjects were more concerned about trial quality, future potential benefits (contributing to the medical profession), and the impact on their own health and life. 【Conclusion:】 Research institutions should be aimed at different social background population stratification for recruitment, informed consent, personnel management system and the standard operating procedure, and pay attention to the subjects’ psychological and physiological needs from several aspects to standardize the management of subjects, improve the compliance of subjects to participate in the trial and the quality of clinical trials, and give more humanistic care and help to subjects.

D-RADA16-RGD-Reinforced Nano-Hydroxyapatite/Polyamide 66 Ternary Biomaterial for Bone Formation

BACKGROUND: Nano-hydroxyapatite/polyamide 66 (nHA/PA66) is a composite used widely in the repair of bone defects. However, this material is insufficient bioactivity. In contrast, D-RADA16-RGD self-assembling peptide (D-RADA16-RGD sequence containing all D-amino acids is Ac-RADARADARADARADARGDS-CONH2) shows admirable bioactivity for both cell culture and bone regeneration. Here, we describe the fabrication of a favorable biomaterial material (nHA/PA66/D-RADA16-RGD). METHODS: Proteinase K and circular dichroism spectroscopy were employed to test the stability and secondary structural properties of peptide D-RADA16-RGD respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the surface of these materials. Confocal laser scanning (CLS), cell counting kit-8 tests (CCK-8), alizarin red S staining, cell immunofluorescence analysis and Western blotting were involved in vitro. Also biosafety and bioactivity of them have been evaluated in vivo. RESULTS: Proteinase K and circular dichroism spectroscopy demonstrated that D-RADA16-RGD in nHA/PA66 was able to form stable-sheet secondary structure. SEM and TEM showed that the D-RADA16-RGD material was 7–33 nm in width and 130–600 nm in length, and the interwoven pore size ranged from 40 to 200 nm. CLS suggests that cells in nHA/PA66/D-RADA16-RGD group were linked to adjacent cells with more actin filaments. CCK-8 analysis showed that nHA/PA66/D-RADA16-RGD revealed good biocompatibility. The results of Alizarin-red S staining and Western blotting as well as vivo osteogenesis suggest nHA/PA66/D-RADA16-RGD exhibits better bioactivity. CONCLUSION: This study demonstrates that our nHA/PA66/D-RADA16-RGD composite exhibits reasonable mechanical properties, biocompatibility and bioactivity with promotion of bone formation.

Preliminary study on spinal nerve coding and injury typing / 中华创伤杂志

Currently,there is no study on the unified coding of the spinal nerve and its main branches.The positions of spinal nerve were encoded according to the basic anatomical principles from top to bottom,from inside to outside and from front to back,with reference to AO bone classification and somatic artery coding and injury classification system.The segmental coding of spinal nerves was decided by its branches and running characteristics.The spinal nerve injuries were encoded by a combination of numbers and letters.The first number represented the region,the second represented the injured nerve,and the third represented the specific segment of injured nerve.The injuries of spinal nerve were divided into 5 categories according to severity.The first letter indicated the category of injury,and the second indicated the orientation of the injured nerve.The functional score scale of the innervation area was prepared based on the evaluation of motor function,sensory function and the results of neurophysiological examination.This scale was used to classify the neurological injuries,guide the clinical treatment and evaluate the prognostic outcomes.This coding and classification system can clearly and comprehensively describe the location and type of spinal nerve injuries,and it is convenient for the diagnosis of nervous system damage.It also has important reference value for treatment and prognosis evaluation.

Dermoscopic features and patterns of onychomycosis / 中华皮肤科杂志

Objective To analyze the dermoscopic features and patterns of onychomycosis.Methods From February to July in 2017,hospitalized patients from the Department of Endocrine Dermatology of the People's Hospital of Xuancheng City was enrolled and subjected to direct microscopic examination of fungi in nails,and the patients with positive results for fungi were diagnosed with onychomycosis.And then,dermoscopic photographs of infected nails were taken to analyze the dermoscopic characteristics and patterns of onychomycosis.Results A total of 205 patients with 634 infected nails were investigated.The dermoscopic characteristics on the nail plates included marble-like turbid areas (47.3％,300/634),pigmentation (30.9％,196/634),splinter hemorrhages (18.3％,116/634) and onycholysis (13.2％,84/634).The prevalence rates of marble-like turbid areas,pigmentation and onycholysis all significantly differed between distal and lateral subungual onychomycosis (DLSO) group and total dystrophic onychomycosis (TDO) group (X2 =42.09,31.23,18.19,respectively,all P ＜ 0.01).The dermoscopic characteristics at the free edge of the nails included nail thickening (33.1％,210/634) and subungual deposition of keratin and debris (26.5％,168/634).The TDO group showed significantly higher prevalence rates of nail thickening and subungual deposition of keratin and detritus compared with the DLSO group (x2 =44.3,18.52,respectively,both P ＜ 0.01).Periungual skin dryness and desquamation occurred more frequently in the TDO group than in the DLSO group (X2 =16.07,P ＜ 0.01).Of the 634lesional nails,141 (22.2％) showed a short spiked pattern,210 (33.1％) showed a longitudinal striated pattern,202 (31.9％) showed a linear edge pattern,and 193 (30.4％) showed a distal irregular termination pattern.Conclusion The main characteristic dermoscopic manifestations of onychomycosis are marblelike turbid areas,subungual deposition of keratin and debris,periungual skin dryness and desquamation,and characteristic dermoscopic patterns of onychomycosis are short spiked pattern,longitudinal striated pattern,linear edge pattern and distal irregular termination pattern.

CRISPR/Cas9 therapeutics: a cure for cancer and other genetic diseases.

Cancer is caused by a series of alterations in genome and epigenome mostly resulting in activation of oncogenes or inactivation of cancer suppressor genes. Genetic engineering has become pivotal in the treatment of cancer and other genetic diseases, especially the formerly-niche use of clustered regularly interspaced short palindromic repeats (CRISPR) associated with Cas9. In defining its superior use, we have followed the recent advances that have been made in producing CRISPR/Cas9 as a therapy of choice. We also provide important genetic mutations where CRISPRs can be repurposed to create adaptive immunity to fight carcinomas and edit genetic mutations causing it. Meanwhile, challenges to CRISPR technology are also discussed with emphasis on ability of pathogens to evolve against CRISPRs. We follow the recent developments on the function of CRISPRs with different carriers which can efficiently deliver it to target cells; furthermore, analogous technologies are also discussed along CRISPRs, including zinc-finger nuclease (ZFN) and transcription activator-like effector nucleases (TALENs). Moreover, progress in clinical applications of CRISPR therapeutics is reviewed; in effect, patients can have lower morbidity and/or mortality from the therapeutic method with least possible side-effects.