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.

[Modified Chevron osteotomy in the treatment of moderate and severe hallux valgus].

OBJECTIVE: To investigate the method and clinical effect of modified Chevron osteotomy of the distal end of the first metatarsal in the treatment of moderate and severe hallux valgus. METHODS: From January 2015 to January 2019, 28 patients(30 feet) with moderate and severe hallux valgus were treated with modified Chevron osteotomy combined with lateral soft tissue release of the first metatarsophalangeal joint, including 2 males (2 feet) and 26 females (28 feet). The age ranged from 35 to 74 (57.3±9.3) years;10 feet on the left, 16 feet on the right, 2 cases on both sides(4 feet);the course of disease was 3 to 12 (9.32±3.89) years. The changes of hallux valgus angle(HVA), intermetatarsal angle(IMA) between the first and second metatarsals and distal metatarsal articular angle(DMAA) of the first metatarsal were measured and compared before and 6 months after operation. The American Orthopaedic Foot and Ankle Society(AOFAS) thumb joint scoring system was used to evaluate the curative effect. RESULTS: All 28 patients were followed up for 8 to 16 (11.28±3.42) months. The incision healed well in all patients, and there were no complications such as incision infection and metatarsal head necrosis. The healing time of osteotomy site was 6 to 10(7.12±1.34) weeks. Preoperative HVA, IMA, DMAA and AOFAS were (36.06±6.02) °, (21.78±4.16) °, (8.21±2.65) ° and (52.90±10.97) respectively, at six months after operation, they were (8.87±2.46) °, (11.66±2.84) °, (3.65±1.00) ° and (87.45±10.55) respectively, there was significant difference between preoperative and 6 months after operation(P<0.05). At 6 months after operation, AOFAS score was excellent in 20 feet, good in 7 feet and poor in 3 feet. Among the 3 patients with poor scores, 2 were excellent after revision, and 1 was significantly improved after using custom insoles. CONCLUSION: Modified Chevron can effectively correct HVA, IMA and DMAA and improve functional recovery. The modified Chevron osteotomy increases the moving distance and the contact of the osteotomy surface. It can be fixed with multiple screws, has strong correction ability, and can exercise early. It is one of the optional methods for the treatment of moderate and severe hallux valgus.

[Clinical effect modified Chevron osteotomy combined with lateral tissue loosening in treating mild-moderate hallux valgus through internal signal approach].

OBJECTIVE: To explore clinical effect of modified Chevron osteotomy combined with lateral tissue loosening for the treatment of mild-moderate hallux valgus through internal signal approach. METHODS: From July 2015 to June 2016, 26 patients with mild-moderate hallux valgus treated with modified Chevron osteotomy combined with lateral tissue loosening through internal signal approach, including 2 males and 24 females aged from 45 to 65 years old with an average of(54.6±4.8) years old;the courses of diseases ranged from 1 to 5 months with an average of (7.5±3.3) months. Hallux valgus angle(HVA), inter metatarsal angle(IMA) were measured at 12 months after operation, and AOFAS score was applied to evaluate clinical effect before and after operation. RESULTS: All incisions were healed at stage I. No incision occurred infection, metatarsal necrosis and recurrence of hallux valgus deformity. Two patients occurred skin numbness caused by musculocutaneous nerve injury. Twenty-six patients were followed up from 6 to 12 months with an average of(9.12±2.06) months. HVA, IMA were(30.01±3.71)°, (14.00±1.50)° before operation and(9.41±4.16)°, (7.00±0.60)° after operation, which had significant difference. There was statistical significance in AOFAS score before operation 54.77±9.59 and after operation 92.73±5.47, and 19 cases obtained excellent results and 7 moderate. CONCLUSIONS: Modified Chevron osteotomy combined with full thread headless pressure screw fixation and lateral tissue loosening for the treatment of mild-moderate hallux valgus has advantages of excellent exposure, simple operation, stable fixation, rapid recovery. Akin osteotomy with internal capsulorrhaphy were used with lateral loosening and could recover soft tissue balance between lateral and internal, and could receive satisfied clinical effects.

Peripheral nerve injury decreases the expression of metabolic glutamate receptor 7 in dorsal root ganglion neurons.

Group II and III metabolic glutamate receptors (mGluRs) are responsible for the glutamate-mediated postsynaptic excitation of neurons. Previous pharmacological evidences show that activation of mGluR7 could inhibit nociceptive reception. However, the distribution and expression patterns of mGluR7 after peripheral injury remain unclear. Herein we found that mGluR7 was expressed in the rat peptidergic dorsal root ganglion (DRG) neurons and large neurons, but rarely in isolectin B4 positive neurons. Sciatic nerve ligation experiment showed that mGluR7 was anterogradely transported from cell body to the peripheral site. Furthermore, after peripheral nerve injury, mGluR7 expression was down-regulated in both peptidergic and large DRG neurons. Our work suggests that mGluR7 might be involved in the regulation of pathological pain after peripheral nerve injury.

[Effects of Bushen Zhuangjin Decoction containing serum on the apoptosis of chondrocytes induced by mechanics stimulus].

OBJECTIVE: To study the effects of Bushen Zhuangjin Decoction (BZD) containing serum on the apoptosis of chondrocytes induced by mechanics stimulus. METHODS: The BZD containing serum was extracted. The chondrocyte nutritive media was divided into 3 groups, i.e., the common nutritive medium group, the blank rabbit serum medium group, and the BZD nutritive medium group. The apoptosis of chondrocytes was induced by continuing mechanics stimulus in 24 h. Then the chondrocytes were collected. The apoptosis rate of chondrocytes was determined by flow cytometry. The contents of interleukin 1beta (IL-1beta) and nitric oxide (NO) in the corresponding media were determined. RESULTS: The apoptosis of chondrocytes in the BZD nutritive medium group (19.55 +/- 7.98)% was lower than that of the common nutritive medium group (39.32 +/- 13.45)% and the blank rabbit serum medium group (37.87 +/- 9.67)%, showing statistical difference (P < 0.05). The contents of IL-1beta and NO were also lower in the BZD nutritive medium group with statistical difference when compared with those of the other two groups (P < 0.05). CONCLUSION: BZD containing serum could protect mechanics stimulus induced apoptosis of chondrocytes.

[Analysis of perioperation complications of total hip arthroplasty in treating Crowe type IV developmental dysplasia of the hip].

OBJECTIVE: To evaluate the clinical effects of total hip arthroplasty (THA) for Crowe type IV developmental dysplasia of the hip (DDH) and analyze perioperative complications. METHODS: From March 2000 to March 2010, 19 patients (23 hips, of them, 4 patients with bilateral hips) with Crowe type IV DDH underwent THA. There were 5 males and 14 females, with average age of 61.3 years (ranged, 41 to 72 years). All hips were treated with small acetabular components combined with medial protrusion technique in acetabular reconstruction, as well as subtrochanteric shortening osteotomy in femur. Joint function of hips were evaluated according to Harris scoring. RESULTS: All patients were followed up with an average of 4.2 years (ranged, 1 to 8 years). Postoperative X-ray films showed all acetabular prosthesis were in true acetabulum. No loosening and nonunion were found in all patients. Harris scoring improved from preoperative 34.0 +/- 6.9 to postoperative 85.0 +/- 7.5. Complications occurred in 11 cases in the patients, including femoral split fracture in 3 cases, nerve injury in 3 cases, delayed union in 2 cases, dislocation in 3 cases. CONCLUSION: Total hip arthroplasty using small acetabular component, medial protrusion, femoral subtrochanteric shortening osteotomy technique for the Crowe type IV DDH can effectively restore hip function and leg length. But incidence of complications is high. The long-term follow-up is necessary for further study.

[Surgical treatment and the fixator selection for Pilon fractures].

OBJECTIVE: To explore the internal fixation selection and the operative method of Pilon fracture. METHODS: From Aug. 2004 to Aug. 2007,there were 40 patients with Pilon fractures involving 30 males and 10 females. Thirty cases were closed fracture and 10 were open fracture. Seven patients were treated by emergency operation, 3 patients were debrided and suture at first. Twenty-three patients were treated with open reduction and internal fixation (ORIF),the other seventeen patients were treated with LC-DCP,reconstruct plate for fixation or screw, Kirschner wire combine screw for fixation. RESULTS: All patients were followed up for 6 to 32 months (means 19 months). The time of union of fracture was about 3 to 5 months. The outcome was evaluated according to Baired-Jackson criteria, the results were excellent in 27 cases,good in 7, fair in 5, poor in 1. Seven cases were found the operative incision and wound surface difficult to heal,among them 5 cases came from the emergency operation patients, 1 case was closed fracture patient. They were be healed by change of dressing or dermatoplasty or skin flap. CONCLUSION: It is the key factor of treatment for Pilon fractures to correct evaluate before operation and select correct operation time. It is also important to selected suitable internal fixation and good reduction.

[Spiral CT reconstruction for typing of tibial plateau fracture to guide surgical therapy].

OBJECTIVE: To typing of tibial plateau fracture based on spiral CT reconstruction and to explore effect of the typing method for treatment. METHODS: A hundred and twenty-six cases with tibial plateau fracture (male 95, female 31, age from 23 to 58 years old), the fractures were classified based on reconstruction image of spiral CT. Including central compression type in 13 cases, split type in 8, split compression type in 79, comminution type in 26. According to the different typing the suitable incision of operation and fixed method for fracture were select. RESULTS: A hundred and twenty-six cases were followed up for 0.5-4 years with an average of 1.2 years. According to Hohl system score to knee joint function, there were statistical significance in the pain,active movement,active range of motion between before and after operation (P < 0.01) and there were no statistical significance in stability and self-evaluation (P < 0.01). CONCLUSION: Typing of tibial plateau fracture based on spiral CT reconstruction helpful to choose operative approach, reduction and fixed method and obviously improve clinical effect.