Effect of mutation at c.493T>C locus of transcription factor HNF1α gene on its protein level.

Hepatocyte nuclear factor 1α (HNF1α) is a transcription factor that is crucial for the regulation to maintain the function of pancreatic ß-cell, hepatic lipid metabolism, and other processes. Mature-onset diabetes of the young type 3 is a monogenic form of diabetes caused by HNF1α mutations. Although several mutation sites have been reported, the specific mechanisms remain unclear, such hot-spot mutation as the P291fsinsC mutation and the P112L mutation and so on. In preliminary studies, we discovered one MODY3 patient carrying a mutation at the c.493T>C locus of the HNF1α gene. In this study, we analyzed the pathogenic of the mutation sites by using the Mutation Surveyor software and constructed the eukaryotic expression plasmids of the wild-type and mutant type of HNF1α to detect variations in the expression levels and stability of HNF1α protein by using Western blot. The analyses of the Mutation Surveyor software showed that the c.493T>C site mutation may be pathogenic gene and the results of Western blot showed that both the amount and stability of HNF1α protein expressed by the mutation type plasmid were reduced significantly compared to those by the wild type plasmid (P<0.05). This study suggests that the c.493T>C (p.Trp165Arg) mutation dramatically impacts HNF1α expression, which might be responsible for the development of the disease and offers fresh perspectives for the following in-depth exploration of MODY3's molecular pathogenic process.

The Treatment of Subtrochanteric Fracture with Reversed Contralateral Distal Femoral Locking Compression Plate (DF-LCP) Using a Progressive and Intermittent Drilling Procedure in Three Osteopetrosis Patients.

OBJECTIVE: To describe the application of reversed contralateral distal femoral locking compression plate (DF-LCP) inserted through a progressive and intermittent drilling procedure in the treatment of osteopetrotic subtrochanteric fracture (OSF). METHODS: Three patients (one male and two females with an average age of 45.33 ± 11.09 years) with OSF hospitalized between September 2015 and September 2020, were included in this present study. Lateral approach was applied in all patients who accepted open reduction and internal fixation (ORIF) with a reversed contralateral DF-LCP inserted through a progressive and intermittent drilling procedure. The operation time and intraoperative blood loss were recorded to evaluate the efficiency of this surgical method. Physical examination and imaging examination of the fracture site were used to evaluate the fracture union status, the position and stability of the implant, and the alignment of the injured limb at 1, 3, 6, and 12 months after operation, then a subsequent visit was conducted at least once a year. Harris Hip Score (HHS) was used to evaluate the hip joint function at 6 and 12 months after operation. RESULTS: The average operation time was 140 ± 21.60 min (110, 160, and 150 min); The average intraoperative blood loss was about 333.33 ± 23.57 ml (300, 350, and 350 ml). The average follow-up time was 22.33 ± 7.41 months (29, 26, and 12 months). All patients achieved bone union with an average time of 6.67 ± 0.94 months (6, 8, and 6 months). At the time of 6 months after operation, case 1 and 3 were almost pain-free and could walk with full weight bearing while case 2 could walk only with partial weight bearing using a crutch. The HHS scores of cases 1, 2, and 3 were 84/100, 74/100, and 92/100, respectively. At the follow-up at 12 months after operation, the HHS score improved to 91/100, 81/100, and 96/100, respectively. The contralateral incomplete old subtrochanteric fracture was deteriorated in case 1 at 26 months after operation. After 3 months of limited weight bearing using a crutch, bone union was verified in radiograph imaging. Fresh contralateral subtrochanteric fracture occurred in case 2 at 26 months after operation, which was treated using a similar surgical approach, and its clinical outcome is under follow-up. Moreover, no perioperative complications including operation-related death, vascular/nerve injury, deep venous thrombosis, pulmonary embolism, and incision infection, and long-term complications involving malunion, nonunion, implant failure, ankylosis, heterotopic ossification, osteonecrosis, and osteomyelitis were identified. CONCLUSION: The application of reversed contralateral DF-LCP in OSF is practicable and reliable. Progressive and intermittent drilling is a safe and efficient method for implant insertion in this complicated situation.

Treatment of Chronic Anterior Shoulder Dislocation by Coracoid Osteotomy with or without Bristow-Latarjet Procedure.

OBJECTIVE: To investigate the clinical efficacy and outcomes of the coracoid osteotomy with or without Bristow-Latarjet procedures in the treatment of chronic anterior shoulder dislocation (CASD). METHODS: Between January 2013 and January 2019, 20 shoulders of 18 patients who were diagnosed with chronic anterior dislocation and underwent open reduction in our trauma center were retrospectively studied. Open coracoid osteotomy with Bristow-Latarjet procedures were performed on 16 shoulders and open coracoid osteotomy without Bristow-Latarjet procedures were performed on four shoulders. Open coracoid osteotomy with or without Bristow-Latarjet procedures were chosen on the basis of the stability of the shoulder after reduction. Outcomes were assessed preoperatively and postoperatively with the visual analog scale (VAS) for pain, the American Shoulder and Elbow Surgeons (ASES) score, the University of California Los Angeles (UCLA) shoulder rating scale, and the range of motion (ROM) for shoulder activity. RESULTS: There were three males and 15 females with an average age of 60.94 ± 2.69 years. The time between dislocation and treatment ranged from 21 to 240 days with an average of 73.3 ± 14.4 days. All patients were available for a mean follow-up of 15.2 ± 4.3 months. No procedure-related death or incision-related superficial or deep tissue infection was identified in all cases. No iatrogenic neurovascular injuries or fractures were found in this study. At the time of 12 months follow-up, the range of motion and the shoulder functional evaluation (VAS [P < 0.001], ASES [P < 0.001], and UCLA score [P < 0.001]) in patients who underwent Bristow-Latarjet procedures were significantly improved. Subluxation after surgical procedure was found and confirmed in one patient and this patient refused to undergo revision surgery. According to the Samilson and Prieto classification system, 16 shoulders were assessed as grade 0, three shoulders were grade 1, one shoulder was grade 2. CONCLUSIONS: Coracoid osteotomy with or without Bristow-Latarjet procedure yielded an acceptable clinical result in this study. This method has the advantages of enlarging the exposure of surgical field, assisting reduction of shoulder, and convenient conversion to Bristow-Latarjet procedure. It is an efficient and reliable method for treatment of chronic anterior shoulder dislocation. A 69-year-old woman diagnosed with right chronic anterior shoulder dislocation with large Hill-Sachs lesion. The latarjet procedure with remplissage technique was applied for this patient.

Surgical and Non-Surgical Treatment for 3-Part and 4-Part Fractures of the Proximal Humerus: A Systematic Review of Overlapping Meta-Analyses.

This systematic review of overlapping meta-analyses was conducted to propose a principle to make decisions for comparing clinical safety and efficacy of surgical and non-surgical treatment for displaced 3-part and 4-part fractures of the proximal humerus. Three electronic databases (PubMed, EMBASE, and the Cochrane Library) were systematically searched to retrieve available published systematic reviews and meta-analyses comparing surgical versus non-surgical treatment for displaced 3-part and 4-part fractures of the proximal humerus. Ten meta-analyses were identified for this investigation. Based on the application of selection with the Jadad algorithm, a meta-analysis including 6 randomized clinical trials was selected for this systematic review, which demonstrated the best available evidence that no statistically significant differences were found in the Constant score, health-related quality of life, and mortality between surgical and non-surgical treatments for displaced 3-part and 4-part fractures of the proximal humerus; however, surgical treatment was associated with a significant increase in the incidence of reoperation. This systematic review of  overlapping meta-analyses reveals that although surgical treatment is more advantageous than and superior to non-surgical treatment for displaced 3-part and 4-part fractures of the proximal humerus, the former leads to a higher incidence of postoperative complications.

Retracted: Downregulation of microRNA-367 promotes osteoblasts growth and proliferation of mice during fracture by activating the PANX3-mediated Wnt/ß-catenin pathway.

A majority of people suffering from bone fractures fail to heal and develop a nonunion, which is a challenging orthopedic complication requiring complex and expensive treatment. Previous data showed the inhibition of some microRNAs (miRNAs or miRs) can enhance fracture healing. The objective of the present study is to explore effects of miR-367 on the osteoblasts growth and proliferation of mouse during fracture via the Wnt/ß-catenin pathway by targeting PANX3. Primarily, the femur fracture model was successfully established in 66 (C57BL/6) 6-week-old male mice. To verify whether miR-367 target PANX3, we used the target prediction program and performed luciferase activity determination. Subsequently, to figure out the underlying regulatory roles of miR-367 in fracture, osteoblasts were elucidated by treatment with miR-367 mimic, miR-367 inhibitor, or siRNA against PANX3 to determine the expression of miR-367, siPANX3, ß-catenin, and Wnt5b as well as cell proliferation and apoptosis. The results demonstrated that PANX3 was verified as a target gene of miR-367. MiR-367 was found to highly expressed but PANX3, ß-catenin, and Wnt5b were observed poorly expressed in fracture mice. downregulated miR-367 increased the mRNA and protein expression of PANX3, ß-catenin, and Wnt5b, increased cell growth, proliferation, and migration, while decreased cell apoptosis in osteoblasts. Altogether, our study demonstrates that the downregulation of miR-367 may promote osteoblasts growth and proliferation in fracture through the activation of the PANX3-dependent Wnt/ß-catenin pathway.

Optimization of ultrasound-assisted aqueous two-phase system extraction of polyphenolic compounds from Aronia melanocarpa pomace by response surface methodology.

Aronia melanocarpa berries are abundant in polyphenolic compounds. After juice production, the pomace of pressed berries still contains a substantial amount of polyphenolic compounds. For efficient utilization of A. melanocarpa berries and the enhancement of polyphenolic compound yields in Aronia melanocarpa pomace (AMP), total phenolics (TP) and total flavonoids (TF) from AMP were extracted, using ultrasound-assisted aqueous two-phase system (UAE-ATPS) extraction method. First, the influences of ammonium sulfate concentration, ethanol-water ratio, ultrasonic time, and ultrasonic power on TP and TF yields were investigated. On this basis, process variables such as ammonium sulfate concentration (0.30-0.35 g mL-1), ethanol-water ratio (0.6-0.8), ultrasonic time (40-60 min), and ultrasonic power (175-225 W) were further optimized by implementing Box-Benhnken design with response surface methodology. The experimental results showed that optimal extraction conditions of TP from AMP were as follows: ammonium sulfate concentration of 0.324 g mL-1, ethanol-water ratio of 0.69, ultrasonic time of 52 min, and ultrasonic power of 200 W. Meanwhile, ammonium sulfate concentration of 0.320 g mL-1, ethanol-water ratio of 0.71, ultrasonic time of 50 min, and ultrasonic power of 200 W were determined as optimum extraction conditions of TF in AMP. Experimental validation was performed, where TP and TF yields reached 68.15 ± 1.04 and 11.67 ± 0.63 mg g-1, respectively. Close agreement was found between experimental and predicted values. Overall, the present results demonstrated that ultrasound-assisted aqueous two-phase system extraction method was successfully used to extract total phenolics and flavonoids in A. melanocarpa pomace.

Spatial distribution of polygalacturonase-inhibiting proteins in Arabidopsis and their expression induced by Stemphylium solani infection.

Disease-induced polygalacturonase-inhibiting proteins (PGIPs) are the major defense proteins which play an important role in resistance to infection of pathogens. To date, the AtPGIP expression in Arabidopsis induced by Stemphylium solani (S. solani) was not described. Here the distribution of AtPGIPs and their expression induced by S. solani infection in Arabidopsis was reported. Notably, immunofluorescence localization showed that the AtPGIPs were distributed in leaves, petioles, stems and roots of 5 week old Arabidopsis, but they were mainly localized in epidermis, vascular bundles and vascular cylinder. Further studies indicated that the transcription level of AtPGIP1 and AtPGIP2 was both up-regulated in response to infection with S. solani which caused hypersensitive cell death, but the transcription level of AtPGIP2 was less induced than AtPGIP1. Consistently, the bulk AtPGIPs of Arabidopsis showed a higher activity in leaves infected by S. solani. Taken together, our preliminary results showed that AtPGIPs were spatially distributed and AtPGIP expression might take part in resistance to infection of S. solani. This study might highlight the potential importance of AtPGIPs and plant disease resistance.