Polydatin protects against articular cartilage degeneration by regulating autophagy mediated by the AMPK/mTOR signaling pathway.

BACKGROUND: Knee osteoarthritis (KOA) is one of the leading causes of disability. Polydatin has a potential effect on KOA treatment but the therapeutic mechanism is not clear. This study aims to investigate the therapeutic action of polydatin in KOA and its mechanism in activating autophagy via the AMP-activated protein kinase (AMPK)/mTOR signaling pathway. METHODS: After a KOA rat model was established by anterior cruciate ligament transection surgery, model rats were treated with polydatin 40 mg/kg for 30 days. Subsequently, cartilage tissues were collected, and hematoxylin-eosin (HE), Safranin-O, and TUNEL staining, and western blotting were performed to evaluate the pathological damage and autophagy-related protein expression. Then, human chondrocyte C28/I2 cells were stimulated with lipopolysaccharide (LPS), and the effects of polydatin on C28/I2 cell viability, apoptosis, and autophagy-related protein expression were detected by MTT, Flow Cytometry, and western blot. In addition, an AMPK inhibitor (Dorsomorphin 2HCl) was used to probe the cell proliferation and apoptosis of polydatin-administered C28/I2 cells. RESULTS: Polydatin ameliorated the pathological damage in rat cartilage tissues and inhibited cell apoptosis in KOA rats. Meanwhile, in C28/I2 cells, polydatin promoted viability and reduced apoptosis. In addition, the protein expression of collagen II, LC3II/LC3I, Beclin-1, and p-AMPK/AMPK were upregulated, and p62 and p-mTOR/mTOR were downregulated by polydatin treatment. Interestingly, relative results showed that the protective effect of polydatin in LPS-stimulated-C28/I2 cells was blocked by the AMPK/mTOR inhibitor, dorsomorphin 2HCl. CONCLUSION: Our research showed that polydatin reduced apoptosis and activated autophagy both in vivo and in vitro by the AMPK/mTOR signaling pathway to protect against KOA, which provided the basis for further investigation into the potential therapeutic impact of polydatin on KOA.

Platelet-rich plasma-derived exosomes promote rotator cuff tendon-bone healing.

BACKGROUND: Rotator cuff tear (RCT) is the most common type of shoulder joint injury, platelet-rich plasma-derived exosomes (PRP-exos) are highly promising in tissue repair and regeneration. The purpose of this study was to determine the function of PRP-exos in rotator cuff tendon-bone healing. METHODS: PRP-exos were isolated from the rabbit whole blood by differential ultracentrifugation and characterized through transmission electron microscopy assay, nanoparticle tracking analysis, and western blotting. Alkaline phosphatase and Von Kossa staining were used to show tendon-derived stem cell (TDSC) differentiation. RT-qPCR and western blotting were performed to detect COL II, SOX-9, and TIMP-1. To determine the therapeutic effects of PRP-exos in vivo. Thirty New Zealand white rabbits were divided into control, model, and PRP-exos groups. The RCT animal model was constructed. The changes in tendon-bone tissue were determined by HE staining. Contents of COL-II, SOX-9, and TIMP-1 were determined by immunohistochemistry staining. RESULTS: PRP-exos were successfully isolated from rabbit blood. PRP-exos promoted TDSC proliferation and differentiation and also induced tendon-specific markers COL II, SOX-9, and TIMP-1 production. In vivo study revealed that PRP-exos promoted early healing of injured tendons. Rabbits treated with PRP-exos had better tissue arrangement in the tear site. Additionally, the contents of COL II, SOX-9, and TIMP-1 were also increased in the RCT rabbit model after PRP-exos treatment. CONCLUSIONS: PRP-exos enhanced tendon-bone healing by promoting TDSC proliferation and differentiation. This finding indicates that PRP-exos can serve as a promising strategy to treat rotator cuff tendon-bone healing.

Total flavonoids of Rhizoma drynariae improves tendon-bone healing for anterior cruciate ligament reconstruction in mice and promotes the osteogenic differentiation of bone mesenchymal stem cells by the ERR1/2-Gga1-TGF-ß/MAPK pathway.

BACKGROUND: Total flavonoids of Rhizoma drynariae (TFRD) is broadly used in the treatment of orthopedic diseases. Nevertheless, the effects and underlying mechanism of TFRD on tendon-bone healing after anterior cruciate ligament reconstruction (ACLR) remain unclear. METHODS: The ACLR mouse model was established. Hematoxylin and Eosin (HE) staining was used for histological analysis of tendon-bone healing. Western blot was utilized to detect the levels of osteogenic related factors (ALP, OCN, RUNX2). The viability and alkaline phosphatase (ALP) activity of bone mesenchymal stem cells (BMSCs) were determined by Cell Counting Kit-8 (CCK-8) and ALP assays. The interaction of estrogen related receptor alpha (ESRRA), estrogen related receptor beta (ESRRB), and golgi-localized γ-ear containing ADP ribosylation factor-binding protein 1 (Gga1) was detected by luciferase reporter assays. The levels of important proteins on the TGF-ß/MAPK pathway were measured by western blot. RESULTS: TFRD improved tendon-bone healing, restored biomechanics of ACLR mice and activated the TGF-ß/MAPK pathway. TFRD treatment also enhanced the viability and osteogenic differentiation of BMSCs in vitro. Then, we demonstrated that TFRD targeted ESRRA and ESRRB to transcriptionally activate Gga1 expression. Knockdown of ESRRA, ESRRB, or Gga1 suppressed the viability and osteogenic differentiation of TFRD-induced BMSCs, which was revealed to be restored by Gga1 overexpression. The overexpression of ESRRA, ESRRB, or Gga1 was demonstrated to promote the BMSC viability and osteogenic differentiation. TGF-ß1 treatment can reverse the impact of Gga1 inhibition on osteogenic differentiation in TFRD-induced BMSCs. CONCLUSION: TFRD improves tendon-bone healing in ACLR mouse models and facilitates the osteogenic differentiation of BMSCs through the ERR1/2-Gga1-TGF-ß/MAPK pathway, which might deepen our understanding of the underlying mechanism of TFRD in tendon-bone healing.

Application of Transumbilical Laparoscopic Surgery on Low/Ultralow Rectal Cancer for Anal Sphincter Preservation.

Background: Rectal cancer is a common malignant tumor in the gastrointestinal tract. This work compares the effects of transumbilical laparoscopic surgery (TULS) and laparoscopic-assisted surgery on the anus-preserving effect of low/ultralow rectal cancer. Materials and Methods: Eighty patients with rectal cancer admitted to our hospital from February 2011 to July 2016 were randomly selected and divided into the laparoscopic group and TULS group, 40 cases in each group, all underwent radical anorectal cancer radical surgery. Statistical analysis was performed on surgical-related indicators in the two groups. Results: Two patients converted to open surgery were excluded. Five patients were excluded because of radical abdomen perineal resection for rectal cancer. Six patients were converted to TULS from laparoscopic surgery. Sixty-seven patients in the experimental group successfully completed anus-sparing surgery, and none died during the operation. The compliance rate of the distance between the lower edge of the tumor and the incision edge of the specimen in the TULS group was better than that in the laparoscopic group (P < .05). There were no significant differences between the two groups in terms of surgical time, blood loss, number of lymph node dissections, functional time of voluntary defecation and postoperative complications, tumor-free recurrence rate at 3 years, and 3-year survival rate after surgery (P > .05). Conclusions: The TULS method is safe and feasible in low and ultralow rectal cancer surgery. It has more advantages than laparoscopic-assisted surgery for anus preservation.

Chondroprotective Effect of Wufu Decoction on Tumor Necrosis Factor-α-Induced Chondrocytes via the Extracellular Signal Regulated Kinase 1/2 Signaling Pathway.

OBJECTIVE: Wufu Decoction (WFD) is a herbal formulation composed of five traditional Chinese herbs that is used clinically for arthritis treatment in China. The current study investigated the chondroprotective effects and the underlying mechanism of WFD for osteoarthritis (OA) therapy. METHODS: The chondroprotective effects of WFD were investigated based on vitro study. Following the successful isolation of chondrocytes from rat cartilage tissues and the identification of collagen II expression with immunofluorescence staining, chondrocytes were co-incubated with tumor necrosis factor-α (TNF-α) to induce an inflammation model; WFD was also administered. After the treatment, cell viability was determined by MTT assay, cell apoptosis was assessed by DAPI staining, the concentration of inflammation cytokines interleukin (IL)-1ß and IL-6 were detected with ELISA assay, the expression of collagen II, MEK1/2-ERK1/2 signaling pathway proteins was detected using western blotting, and mRNA expression of MMP-1, MMP-9 and MMP-13 were determined with quantitative real-time polymerase chain reaction. RESULTS: Wufu Decoction significantly restored the cell viability suppressed by TNF-α and inhibited the cell apoptosis induced by TNF-α in chondrocytes. The high concentrations of IL-1ß and IL-6 in TNF-α-induced model cells were significantly decreased in WFD-treated chondrocytes, and the immunofluorescence staining and western blot results showed that the inhibited expression of collagen II in the TNF-α-induced model group was significantly increased in WFD-treated chondrocytes. The protein expressions of MEK1/2, p-ERK1/2, and P53 were significantly reduced in the WFD-treated group compared with those in the model group, and the mRNA expressions of MMP-1, MMP-9, and MMP-13 were also significantly reduced with WFD treatment. CONCLUSION: The present study indicated that WFD exerted a chondroprotective effect in TNF-α-induced chondrocytes via the regulation of the ERK1/2 signaling pathway, suggesting that WFD has therapeutic potential for OA therapy.

[Reverse total shoulder arthroplasty for the treatment of old fracture of proximal humerus in elderly patients].

OBJECTIVE: To investigate clinical results of reverse total shoulder arthroplasty in treating old fracture of proximal humerus in elderly patients. METHODS: From January 2012 to December 2017, 12 elderly patients with old proximal humeral fractures were treated with reverse total shoulder arthroplasty. There were 5 males and 7 females with an average age of 70.2 years old (ranged from 63 to 81 years old) and an average course of 12.3 months (ranged from 9 to 18 months). VAS score was used to evaluate the degree of pain relief of shoulder joint, Constant-Murley score was used to evaluate the improvement of shoulder joint function and observe the complications during and after operation. RESULTS: All the 12 patients were followed up with an average duration of 22.3 months (ranged from 15 to 56 months). At the latest follow-up, the VAS score was 1.9±1.4, and Constant-Murley score was 83.4±8.4. Among them, shoulder joint flexion was (92.5±7.7)°, abduction was (90.4±14.3)°, external rotation was(31.9±10.0)°, and internal rotation was(58.9±13.1)°. There was statistical significance before and after treatment(P<0.05). There were 2 cases with glenoid notch. The imaging findings were grade I and grade II respectively. No loosening was found during follow-up. No infection, acromial fracture, prosthesis loosening, axillary nerve injury and other complications occurred. CONCLUSIONS: Reverse total shoulder arthroplasty in treating old proximal humeral fractures in elderly patients can achieve satisfactory results, however, indications and complications should be noted.

[Comparison of long Gamma 3 nail and proximal femur locking plate for the treatment of femoral subtrochanteric fractures].

OBJECTIVE: To compare clinical efficacy of long Gamma 3 nail and proximal femur locking plate (PFLP) in treating femoral subtrochanteric fractures. METHODS: From January 2010 to January 2017, clinical data of 58 patients with subtrochanteric fractures followed more than 12 months were retrospective analyzed. Among them, 35 patients were treated with long Gamma 3 nail including 18 males and 17 females aged from 25 to 78 years old with an average of(66.5±23.5) years old;Causes of injury included fall on the ground in 18 cases, traffic accidents in 7 cases, and fall from height in 10 cases. The other 23 patients were treated with PFLP fixation including 8 males and 15 females aged from 31 to 81 years old with an average of (63.4±22.4) years old;Causes of injury included fall on the ground in 12 cases, traffic accidents in 6 cases, and fall from height in 5 cases. Operative time, blood loss (intraoperative and hidden blood loss), hospital stays, bone healing and complications were observed and compared. Harris hip score after 1-year following-up was used to evaluate postoperative clinical effect. RESULTS: Fifty-one patients were followed up from 14 to 36 months with an average of 24.8 months, including 31 patients were treated with long Gamma 3 and 20 patients were treated with PFLP. Blood loss(intraoperative and hidden blood loss) in PFLP group was less than that of long Gamma 3 nail group(P<0.05). There was no significant in operative time, hospital stays and complications between two groups(P>0.05). There was no significant differences in healing time of fractures between long Gamma 3 nail group (17.2±2.4) weeks and PFLP group (18.1±2.6) weeks(P<0.05). At 1-year following-up, there was no significant differences in Harris hip score between long Gamma 3 nail group(80.29±10.28) and PFLP group (76.49±12.28)(P<0.05). No complications such as pulmonary embolism and nonunion occurred. Two patients were treated with fitler whose occurred deep vein thrombosis. Postoperative pulmonary infection curred in 4 cases and was cured by anti-infection therapy. CONCLUSIONS: Both of long Gamma 3 nail and PFLP in treating patients with femoral subtrochanteric fractures can receive good clinical effects, long Gamma 3 nail is not suitable for the patients of the narrow medullary cavity and prominent anterior arch. PFLP is eccentric fixation, so early weight-bearing was not stress.