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.

[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.