Investigating the synergy of Shikonin and Valproic acid in inducing apoptosis of osteosarcoma cells via ROS-mediated EGR1 expression.

BACKGROUND: Osteosarcoma is the most prevalent malignant bone tumour with a poor prognosis. Shikonin (SHK) is derived from the traditional Chinese medicine Lithospermum that has been extensively studied for its notable anti-tumour effects, including for osteosarcoma. However, its application has certain limitations. Valproic acid (VPA) is a histone deacetylase inhibitor (HDACI) that has recently been employed as an adjunctive therapeutic agent that allows chromatin to assume a more relaxed state, thereby enhancing anti-tumour efficacy. PURPOSE: This study was aimed to investigate the synergistic anti-tumour efficacy of SHK in combination with VPA and elucidate its underlying mechanism. METHODS/STUDY DESIGN: CCK-8 assays were utilized to calculate the combination index. Additional assays, including colony formation, acridine orange/ethidium bromide double fluorescent staining, and flow cytometry, were employed to evaluate the effects on osteosarcoma cells. Wound healing and transwell assays were utilized to assess cell mobility. RNA sequencing, PCR, and Western blot analyses were conducted to uncover the underlying mechanism. Rescue experiments were performed to validate the mechanism of apoptotic induction. The impact of SHK and VPA combination treatment on primary osteosarcoma cells was also assessed. Finally, in vivo experiments were conducted to validate its anti-tumour effects and mechanism. RESULTS: The combination of SHK and VPA synergistically inhibited the proliferation and migration of osteosarcoma cells in vitro and induced apoptosis in these cells. Through a comprehensive analysis involving RNA sequencing, PCR, Western blot, and rescue experiments, we have substantiated our hypothesis that the combination of SHK and VPA induced apoptosis via the ROS-EGR1-Bax axis. Importantly, our in vivo experiments corroborated these findings, demonstrating the potential of the SHK and VPA combination as a promising therapeutic approach for osteosarcoma. CONCLUSION: The combination of SHK and VPA exerted an anti-tumour effect by inducing apoptosis through the ROS-EGR1-Bax pathway. Repurposing the old drug VPA demonstrated its effectiveness as an adjunctive therapeutic agent for SHK, enhancing its anti-tumour efficacy and revealing its potential value. Furthermore, our study expanded the application of natural compounds in the anti-tumour field and overcame some of their limitations through combination therapy. Finally, we enhanced the understanding of the mechanistic pathways linking reactive oxygen species (ROS) accumulation and apoptosis in osteosarcoma cells. Additionally, we elucidated the role of EGR1 in osteosarcoma cells, offering novel strategies and concepts for the treatment of osteosarcoma.

Age differences in the impact of dietary salt on metabolism, blood pressure and cognitive function in male rats.

The influence of salt consumption on physiological processes, especially blood pressure (BP), metabolism, and cognition, remains a topical concern. While guidelines endorse reduced salt diets, there are gaps in understanding the age-specific implications and challenges in adherence. The present study delved into the differential effects of salt intake on young adult and aged male rats over a 12-week period, using control, low-, and high-salt diets. Key metrics, such as BP, cognition, and general parameters, were monitored. Our findings revealed significant age-dependent effects of salt intake on survival rates, body weight, blood sodium, blood glucose, blood lipids, BP, heart rates, and cognition. Notably, young adult rats did not show significant sodium level changes on a high-salt diet, whereas aged rats experienced increased sodium levels even on a normal salt diet. Blood glucose levels decreased significantly in aged rats on a high-salt diet but remained stable in young adults. Aged rats had the highest survival rates on low-salt diets. Low-salt diets led to reduced BP in both age groups, more significantly in young adults. Young adult rats displayed increased BP variability on both high- and low-salt diets, while a decrease in BP variability was exclusive to aged rats on a low-salt diet. There were significant differences across age groups in short-term memory, but not in long-term memory. The study provides a nuanced understanding of the age-dependent physiological effects of salt intake, suggesting the necessity of age-specific guidelines for public health.

Review: Predictive approaches to breast cancer risk.

Despite the deployment of specific breast cancer screening strategies, breast cancer incidence rates have escalated significantly over recent decades. In a bid to reverse this trend, scientists have engaged in extensive epidemiological research into breast cancer prevalence, identifying numerous individual risk factors and promoting population-wide health education. Coupled with advances in genetic testing, risk prediction models based on breast cancer genes have been developed, albeit with inherent limitations. In the new millennium, the emergence of artificial intelligence (AI) as a dominant technological force suggests that breast cancer prediction models developed with AI may represent the next frontier in research.

[Effect mechanism of pricking and cupping therapy combined with Chinese herbal wet compress in treatment of herpes zoster based on Th1/Th2 imbalance and serum substance P].

OBJECTIVE: To observe the effects of pricking and cupping therapy combined with Chinese herbal wet compress on Th1/Th2 balance and the concentration of substance P (SP) in treatment of patients with herpes zoster at acute stage and explore the mechanism of clinical efficacy of this combined therapeutic method. METHODS: The patients of herpes zoster at acute stage were randomly divided into a treatment group (58 cases), a control group No.1 (57 cases), a control group No.2 (58 cases) and a control group No.3 (59 cases). In the treatment group, the therapeutic regimen was pricking and cupping+Chinese herbal wet compress+basic treatment. In the control group No.1, pricking and cupping + basic treatment were administered. In the control group No.2, Chinese herbal wet compress+basic treatment were provided. In the control group No.3, only basic treatment was delivered. The treatment duration was 9 days in each group. The score of pain degree, the score of sleep quality and the rehabilitation conditions of pain and herpes were observed in 4 groups. The concentration of serum interleukin 4 (IL-4), interferon gamma (IFN-γ) and SP were detected by ELISA. The clinical efficacy was evaluated. The occurrence of adverse reaction during treatment was observed. RESULTS: After treatment, the score of pain degree and the concentrations of serum IL-4 and SP were all lower than those before treatment in 4 groups (P<0.05); and the score of subjective sleep quality, the concentrations of serum IFN-γ and IFN-γ/IL-4 were all increased in comparison with those before treatment (P<0.05). In the treatment group and the control groups No.1 and No.2, the score of pain degree and the concentrations of serum IL-4 and SP were all lower than those in the control group No.3 (P<0.05); and the score of subjective sleep quality, the concentrations of serum IFN-γ and IFN-γ/IL-4 were higher than those in the control group No.3, respectively (P<0.05). Besides, every indicator in the treatment group was improved more significantly than that either in the control group No.1 or the control group No.2 (P<0.05). After treatment, compared with the control group No.3, the time of pain relief and disappearance, and the time of blister termination, incrustation and decrustation were all shorter in the treatment group, the control group No.1 and the control group No.2 separately (P<0.05); and every indicator in the treatment group was reduced more significantly in comparison with either the control group No.1 or the control group No.2 (P<0.05). The total effective rate in the treatment group (96.55%, 56/58), the control group No.1 (92.98%, 53/57) and the control group No.2 (91.38%, 53/58) was higher than that in the control group No.3 (74.58%, 44/59, P<0.05). No any adverse reaction occurred in the 4 groups. CONCLUSION: The pricking and cupping therapy combined with Chinese herbal wet compress effectively promotes the recovery of Th1/Th2 balance and reduces the concentration of serum SP in patients with herpes zoster at acute stage. This combined therapeutic regimen is conductive to the recovery of patients, the improvement of therapeutic effects and the decrease of the risk of pos-therpetic neuralgia.

[Fire needling stripping after local anesthesia for verruca vulgaris: a multi-center randomized controlled trial].

OBJECTIVE: To compare the clinical therapeutic effect of fire needling stripping after local anesthesia, simple fire needling and liquid nitrogen cryotherapy on verruca vulgaris. METHODS: A total of 900 patients with verruca vulgaris were randomized into a fire needling stripping group (300 cases, 2 cases dropped off), a fire needling group (300 cases, 4 cases dropped off) and a liquid nitrogen cryotherapy group (300 cases, 5 cases dropped off). After local anesthesia of compound lidocaine cream, fire needling therapy was adopted, and the necrotic tissue of verruca was stripped in the fire needling stripping group. Simple fire needling therapy was adopted in the fire needling group, without local anesthesia and stripping. Liquid nitrogen cryotherapy was adopted in the liquid nitrogen cryotherapy group. The treatment was given once a week, and totally 3 weeks were required in the 3 groups. The skin lesion scores of number, area, thickness, color, pruritus, isomorphism and the level of T lymphocyte (CD+3、CD+4、CD+8、CD+4/ CD+8) in peripheral blood were observed before and after treatment, and the adverse reaction was recorded in the 3 groups. Five weeks after treatment, the therapeutic effect was evaluated. RESULTS: Compared before treatment, the skin lesion scores were decreased (P<0.05), the levels of T lymphocyte in peripheral blood were increased in the 3 groups (P<0.05). After treatment, all the items of the skin lesion score in the fire needling stripping group were lower than those in the fire needling group and the liquid nitrogen cryotherapy group (P<0.05), the levels of T lymphocyte in peripheral blood were higher than those in the fire needling group and the liquid nitrogen cryotherapy group (P<0.05); all the items of the skin lesion score in the liquid nitrogen cryotherapy group were lower than those in the fire needling group (P<0.05). At the follow-up, the total effective rate was 88.6% (264/298) in the fire needling stripping group, which was superior to 81.4% (241/296) in the fire needling group and 81.4% (240/295) in the liquid nitrogen cryotherapy group (P<0.05). The cases of infection, causalgia and cicatrix in the liquid nitrogen cryotherapy group were more than those in the fire needling stripping group and the fire needling group (P<0.05). CONCLUSION: Fire needling stripping after local anesthesia can effectively treat the verruca vulgaris, improve the skin lesion and immunity, its therapeutic effect is superior to simple fire needling and liquid nitrogen cryotherapy.

Lipoxin A4 inhibits lipopolysaccharide-induced production of inflammatory cytokines in keratinocytes by up-regulating SOCS2 and down-regulating TRAF6.

Liopxin A4 (LXA4) is considered to be a crucial modulator in the inflammatory responses. In the present study, we aimed to study the effect of LXA4 on the inflammatory cytokines production induced by lipopolysaccharide (LPS) and the possible mechanism in normal human epidermal keratinocytes (NHEKs). NHEKs were isolated and cultured. The expression of toll-like receptor 4 (TLR4), LXA4 receptor (ALXR) and aryl hydrocarbon receptor (AhR) in NHEKs was detected by reverse transcription polymerase chain reaction (RT-PCR). The mRNA and protein levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1ß (IL-1ß) were determined in NHEKs stimulated by LPS (10 µg/mL) with or without preincubation with LXA4 (100 nmol/L) for 30 min by real-time quantitative PCR (real-time qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The expression levels of tumor necrosis factor receptor-associated factor 6 (TRAF6) and suppressors of cytokine signaling 2 (SOCS2) mRNAs and proteins, and nuclear translocation of NF-kB-p65 were measured by real-time qPCR and Western blotting, respectively. The results showed that NHEKs expressed TLR4, ALXR and AhR. LXA4 significantly inhibited the mRNA and protein expression levels of TNF-α, IL-1ß and TRAF6 induced by LPS in NHEKs, and LXA4 obviously increased the expression of SOCS2 at mRNA and protein levels. The nuclear NF-kB-p65 protein expression induced by LPS was inhibited after preincubation with LXA4 in NHEKs. It was concluded that LXA4 inhibits the LPS-induced production of TNF-α and IL-1ß in NHEKs by up-regulating SOCS2 and down-regulating TRAF6.

Lipoxin A4 inhibits proliferation and inflammatory cytokine/chemokine production of human epidermal keratinocytes associated with the ERK1/2 and NF-κB pathways.

BACKGROUND: Current in vitro studies show that lipoxin A4 (LXA4) has multiple biological functions including inhibiting cell proliferation and inflammatory cytokine production. Our previous studies showed LXA4 could inhibit the expression of IL-6 and IL-8 in normal human epidermal keratinocytes (NHEKs). However, more specific effects including regulation of cell proliferation and anti-inflammatory mechanisms of LXA4 in NHEKs have not been previously studied. OBJECTIVE: We proposed to investigate the effects of LXA4 on cell proliferation and inflammatory cytokine/chemokine production in NHEKs, and the possible molecular mechanisms of cell cycle and anti-inflammatory signal transduction pathway. METHODS: NHEKs were stimulated with LPS, with or without preincubation with LXA4. Cell proliferation and cell cycle of NHEKs were examined by WST-8, CFSE assay and DNA staining, respectively. The mRNA and protein levels of inflammatory cytokines were quantified by real-time quantitative PCR and ELISA. The expressions of signaling proteins cyclin D1, P16INK4A, ERK1/2 and NF-κB-p65 were analyzed using Western blotting. RESULTS: Cell proliferation and inflammatory cytokine/chemokine production of NHEKs were suppressed by LXA4, which caused G0/G1 phase cell cycle arrest in NHEKs. The expression of cyclin D1 was down-regulated by LXA4, contrary to the results of P16INK4A. The ERK1/2 phosphorylation and NF-κB-p65 nuclear translocation of NHEKs were both suppressed by LXA4. CONCLUSION: Cell growth and inflammatory cytokine/chemokine production of NHEKs were inhibited by LXA4, and the inhibitory effects might be associated with the mechanisms of cyclin D1/P16INK4A, ERK1/2 and NF-κB signal transduction pathway.

[Meta-analysis of studies on mortality of early surgery vs. delayed surgery for patients with femoral neck fractures].

OBJECTIVE: Current guidelines indicate that surgery for femoral neck fracture should be performed within 24 hours. But the effect of early surgery on mortality was not consistent with each other. This study is to explore the effect of early surgery on the risk of death among elderly patients with femoral neck fracture. METHODS: By searching Medline, PubMed, CENTRAL (the Cochrane central register of controlled trials), CBM, CNKI and VIP et al, published documents about studies of early surgery VS delayed surgery on mortality for femoral neck fractures were collected. The data were analysised by using statistic software Stata 11.0. Random or fixed effect models were applied to conduct meta analysis on the study results, the combined relative risk (RR) and the 95% confidence interval (CI) were calculated as well. RESULTS: Six cohort studies and 8 430 patients were included in the combined analysis,which indicated that 64% death risk was observed in the early surgery group compared to the delayed surgery group (RR = 0.64, 95% CI: 0.55-0.73, P = 0.000). The death was divided into two subgroups: one was early death with 30 days and the other was late death after 6 months. For the early death, there was no statistic significance between early surgery and delayed surgery (RR = 0.93, 95% CI: 0.69-1.18, P = 0.076). And for the late death, only 59% death risk was observed in the early surgery group (RR = 0.59, 95% CI: 0.49-0.69, P = 0.000) compared to the delayed surgery group. CONCLUSION: Early surgery was associated with a lower risk of death for femoral neck fracture patients.

[Micro machining of micro-cantilever probes for efficient deposition for biochips].

Biochip technology will bring a tremendous revolution to life science and medical research in 21 century. Microarray assays represent an essential technical advance in biomedical research. Recently, the demand for microarray assay technology has spring up. Therefore, low cost and flexible techniques are needed to meet specific requirements for increasingly integrated biochips. Also performance must be improved in terms of speed and sensitivity. To this end, promising approaches, mainly based on micro and nanotechnologies, have been developed. In this paper, the design and microfabrication of a novel type of micro-cantilever probe are introduced. These probes were fabricated using silicon dioxide by Micro-electromechanical System (MEMS) techniques, and they featured one micron split gap, microchannels and self-replenishing reservoirs. All fabricated micro-cantilever probe were tested on Nanoarrayer instrumentation. Cy3-streptavidin was loaded as biological sample and patterned on DSU gold surface. Results showed these probes were capable of generating high quality biological arrays with routine spot sizes of 2 - 3 microns and could deposit at least three thousand spots without reloading. The spot size could potentially achieve sub-micron when probe size was further shrunk down by the high-resolution lithography technique or more precise microfabrication technologies, such as E-beam lithography. To further improve sample loading efficiency, it is needed to modify the cantilever surface in order to better confine sample inside the microchannel and reservoir, which will be researched in the future.