High-resolution epitope mapping of commercial antibodies to ANCA antigens by yeast surface display.

Epitope mapping provides critical insight into antibody-antigen interactions. Epitope mapping of autoantibodies from patients with autoimmune diseases can help elucidate disease immunogenesis and guide the development of antigen-specific therapies. Similarly, epitope mapping of commercial antibodies targeting known autoantigens enables the use of those antibodies to test specific hypotheses. Anti-Neutrophil Cytoplasmic Autoantibody (ANCA) vasculitis results from the formation of autoantibodies to multiple autoantigens, including myeloperoxidase (MPO), proteinase-3 (PR3), plasminogen (PLG), and peroxidasin (PXDN). To perform high-resolution epitope mapping of commercial antibodies to these autoantigens, we developed a novel yeast surface display library based on a series of >5000 overlapping peptides derived from their protein sequences. Using both FACS and magnetic bead isolation of reactive yeast, we screened 19 commercially available antibodies to the ANCA autoantigens. This approach to epitope mapping resulted in highly specific, fine epitope mapping, down to single amino acid resolution in many cases. Our study also identified cross-reactivity between some commercial antibodies to MPO and PXDN, which suggests that patients with apparent autoantibodies to both proteins may be the result of cross-reactivity. Together, our data validate yeast surface display using maximally overlapping peptides as an excellent approach to linear epitope mapping.

Oxidative Stress Contributes to Slit Diaphragm Defects Caused by Disruption of Endocytosis.

Introduction: Podocyte slit diaphragms are an important component of the glomerular filtration barrier. Podocyte injury frequently includes defects in slit diaphragms, and various mechanisms for these defects have been described, including altered endocytic trafficking of slit diaphragm proteins or oxidative stress. However, the potential relationship between endocytosis and oxidative stress in the context of slit diaphragm integrity has not been extensively considered. Methods: To examine the potential relationships between endocytosis, oxidative stress, and slit diaphragm integrity, we induced genetic or pharmacological disruption of endocytosis in Drosophila nephrocytes (the insect orthologue of podocytes) and cultured human podocytes. We then employed immunofluorescence microscopy to analyze protein localization and levels, and to quantify signal from reactive oxygen species (ROS) dyes. Immunoprecipitation from podocyte cell lysates was used to examine effects on slit diaphragm protein complex formation (i.e., nephrin/podocin and nephrin/ZO-1). Results: Disruption of endocytosis in nephrocytes and podocytes led to slit diaphragm defects, elevated levels of ROS (oxidative stress), and activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant pathway. In nephrocytes with defective endocytosis, perturbation of Nrf2 signaling exacerbated slit diaphragm defects. Conversely, overexpression of Nrf2 target genes catalase or glucose-6-phosphate dehydrogenase (G6PD) significantly ameliorated slit diaphragm defects caused by disruption of endocytosis. Conclusion: Oxidative stress is an important consequence of defective endocytosis and contributes to the defects in slit diaphragm integrity associated with disruption of endocytic trafficking.

Donor-Derived Engineered Microvessels for Cardiovascular Risk Stratification of Patients with Kidney Failure.

Cardiovascular disease is the cause of death in ≈50% of hemodialysis patients. Accumulation of uremic solutes in systemic circulation is thought to be a key driver of the endothelial dysfunction that underlies elevated cardiovascular events. A challenge in understanding the mechanisms relating chronic kidney disease to cardiovascular disease is the lack of in vitro models that allow screening of the effects of the uremic environment on the endothelium. Here, a method is described for microfabrication of human blood vessels from donor cells and perfused with donor serum. The resulting donor-derived microvessels are used to quantify vascular permeability, a hallmark of endothelial dysfunction, in response to serum spiked with pathophysiological levels of indoxyl sulfate, and in response to serum from patients with chronic kidney disease and from uremic pigs. The uremic environment has pronounced effects on microvascular integrity as demonstrated by irregular cell-cell junctions and increased permeability in comparison to cell culture media and healthy serum. Moreover, the engineered microvessels demonstrate an increase in sensitivity compared to traditional 2D assays. Thus, the devices and the methods presented here have the potential to be utilized to risk stratify and to direct personalized treatments for patients with chronic kidney disease.

Effects of microbial agents and corn protein ferment on physiological characteristics in leaves and yield of tomato.

Premature senescence in greenhouse tomato is a significant challenge under long-season cultivation, due to suboptimal nutrient management during growth periods. We investigated the effects of microbial agents (T1), corn protein ferment (T2), and their combined application (T3) on photosynthetic characteristics and antioxidant enzyme activities in 'Saint Laurent 3689' tomato leaves, normal management served as the control (CK). We explored the physiological mechanism of delaying leaf senescence. Results showed that applying microbial agents or corn protein ferment individually led to improvements in leaf photosynthetic characteristics and antioxidant enzyme activities. The combined application yielded superior outcomes. Eighty days post the combined application of microbial agents and corn protein ferment (T3), chlorophyll (a+b) content, net photosynthetic rate, and the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in leaves increased by 16.4%, 30.9%, 23.4%, 33.0% and 40.3%, respectively, compared with the CK. Furthermore, plant height and stem diameter increased by 8.2% and 7.0%, while the total yield exhibited a significant increase of 9.9% compared with the CK 210 days post-treatment. In conclusion, the combined application of microbial agents and corn protein ferment has promising potential in enhancing chlorophyll content, net photosynthetic rate, and the activities of SOD, POD and CAT in tomato leaves. This approach effectively delayed leaf senescence, thereby promoting tomato growth and remarkably increasing the yield.

Prognostic value of quantitative flow ratio in patients with coronary heart disease after percutaneous coronary intervention therapy: a meta-analysis.

Background: Coronary atherosclerotic heart disease is one of the most serious health and life-threatening diseases. There is no doubt that despite the increasing number of assessment methods used clinically, the prognosis assessment is still not ideal, and newer assessment methods are needed. Objective: To investigate the predictive value of quantitative flow ratio (QFR) for adverse events (vessel-oriented composite endpoint events/target lesion failure) in patients after percutaneous coronary intervention (PCI). Method: Eight studies involving 4,173 patients (5,688 vascular lesions) were included. These are studies on the relationship between QFR values and prognosis of adverse cardiac events after PCI. This meta-analysis was performed after quality assessment and data extraction of clinical trials data that met the inclusion criteria. Result: Each of the eight studies described the cut-off values for the best predictive ability of post-PCI QFR and the hazard ratio (HR) between QFR values and adverse events, respectively. The pooled HR of these studies was 4.72 (95% CI: 3.29-6.75). Concurrently, lower post-PCI QFR values were associated with the occurrence of individual clinical events (cardiac death/myocardial infarction/target vessel revascularization), with relative risk values of 6.51 (95% CI: 4.96-8.53), 4.83 (95% CI: 3.08-7.57), and 4.21 (95% CI: 2.66-6.68), respectively. Conclusion: QFR may have great potential in the assessment of prognosis. It is necessary to measure QFR value after PCI. A lower QFR value after PCI was an important predictor for experiencing adverse events.

Development and validation of a postoperative pulmonary infection prediction model for patients with primary hepatic carcinoma.

BACKGROUND: There are factors that significantly increase the risk of postoperative pulmonary infections in patients with primary hepatic carcinoma (PHC). Previous reports have shown that over 10% of patients with PHC experience postoperative pulmonary infections. Thus, it is crucial to prioritize the prevention and treatment of postoperative pulmonary infections in patients with PHC. AIM: To identify the risk factors for postoperative pulmonary infection in patients with PHC and develop a prediction model to aid in postoperative management. METHODS: We retrospectively collected data from 505 patients who underwent hepatobiliary surgery between January 2015 and February 2023 in the Department of Hepatobiliary and Pancreaticospleen Surgery. Radiomics data were selected for statistical analysis, and clinical pathological parameters and imaging data were included in the screening database as candidate predictive variables. We then developed a pulmonary infection prediction model using three different models: An artificial neural network model; a random forest model; and a generalized linear regression model. Finally, we evaluated the accuracy and robustness of the prediction model using the receiver operating characteristic curve and decision curve analyses. RESULTS: Among the 505 patients, 86 developed a postoperative pulmonary infection, resulting in an incidence rate of 17.03%. Based on the gray-level co-occurrence matrix, we identified 14 categories of radiomic data for variable screening of pulmonary infection prediction models. Among these, energy, contrast, the sum of squares (SOS), the inverse difference (IND), mean sum (MES), sum variance (SUV), sum entropy (SUE), and entropy were independent risk factors for pulmonary infection after hepatectomy and were listed as candidate variables of machine learning prediction models. The random forest model algorithm, in combination with IND, SOS, MES, SUE, SUV, and entropy, demonstrated the highest prediction efficiency in both the training and internal verification sets, with areas under the curve of 0.823 and 0.801 and a 95% confidence interval of 0.766-0.880 and 0.744-0.858, respectively. The other two types of prediction models had prediction efficiencies between areas under the curve of 0.734 and 0.815 and 95% confidence intervals of 0.677-0.791 and 0.766-0.864, respectively. CONCLUSION: Postoperative pulmonary infection in patients undergoing hepatectomy may be related to risk factors such as IND, SOS, MES, SUE, SUV, energy, and entropy. The prediction model in this study based on diffusion-weighted images, especially the random forest model algorithm, can better predict and estimate the risk of pulmonary infection in patients undergoing hepatectomy, providing valuable guidance for postoperative management.

An Effective Surgical Strategy for Anteromedial Osteoarthritis of the Knee with Simultaneous Popliteal Cysts Drainage.

Unicompartmental knee arthroplasty (UKA) is an established treatment option for anteromedial osteoarthritis, and popliteal cysts are a common finding in the knee among patients with chronic osteoarthritis pain. The two are so closely related that popliteal cysts are commonly discovered during the unicompartmental knee arthroplasty preoperative examination. However, only a few reports exist on the management and outcome of popliteal cysts in the patients receiving UKA for knee osteoarthritis (OA) and popliteal cysts. As such, it is crucial to evaluate different treatment strategies and their management of popliteal cysts. In this paper, we evaluate a surgical strategy for patients with knee anteromedial osteoarthritis and symptomatic popliteal cysts. These patients were treated with UKA and internal drainage of the popliteal cyst. The results shown here, spanning 1-year post-operation follow-up, demonstrated that UKA and internal drainage is an effective surgical protocol for treating anteromedial osteoarthritis with symptomatic popliteal cysts.

Pharmacokinetic Analysis of Diosgenin in Rat Plasma by a UPLC-MS/MS Approach.

Diosgenin, a steroidal sapogenin, has attracted attention worldwide owing to its pharmacological properties, including antitumor, cardiovascular protective, hypolipidemic, and anti-inflammatory effects. The current diosgenin analysis methods have the disadvantages of long analysis time and low sensitivity. The aim of the present study was to establish an efficient, sensitive ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach for pharmacokinetic analysis of diosgenin amorphous solid dispersion (ASD) using tanshinone IIA as an internal standard (IS). Male Sprague-Dawley rats were orally administered diosgenin ASD, and orbital blood samples were collected for analysis. Protein precipitation was performed with methanol-acetonitrile (50 : 50, v/v), and the analytes were separated under isocratic elution by applying acetonitrile and 0.03% formic acid aqueous solution at a ratio of 80 : 20 as the mobile phase. MS with positive electron spray ionization in multiple reaction monitoring modes was applied to determine diosgenin and IS with m/z 415.2⟶271.2 and m/z 295.2⟶277.1, respectively. This approach showed a low limit of quantification of 0.5 ng/ml for diosgenin and could detect this molecule at a concentration range of 0.5 to 1,500 ng/ml (r = 0.99725). The approach was found to have intra- and inter-day precision values ranging from 1.42% to 6.91% and from 1.25% to 3.68%, respectively. Additionally, the method showed an accuracy of -6.54 to 4.71%. The recoveries of diosgenin and tanshinone IIA were 85.81-100.27% and 98.29%, respectively, with negligible matrix effects. Diosgenin and IS were stable under multiple storage conditions. Pharmacokinetic analysis showed that the C max and AUC0⟶t of diosgenin ASD were significantly higher than those of the bulk drug. A sensitive, simple, UPLC-MS/MS analysis approach was established and used for the pharmacokinetic analysis of diosgenin ASD in rats after oral administration.

Personal predictive model based on systemic inflammation markers for estimation of postoperative pancreatic fistula following pancreaticoduodenectomy.

BACKGROUND: Postoperative pancreatic fistula (PF) is a serious life-threatening complication after pancreaticoduodenectomy (PD). Our research aimed to develop a machine learning (ML)-aided model for PF risk stratification. AIM: To develop an ML-aided model for PF risk stratification. METHODS: We retrospectively collected 618 patients who underwent PD from two tertiary medical centers between January 2012 and August 2021. We used an ML algorithm to build predictive models, and subject prediction index, that is, decision curve analysis, area under operating characteristic curve (AUC) and clinical impact curve to assess the predictive efficiency of each model. RESULTS: A total of 29 variables were used to build the ML predictive model. Among them, the best predictive model was random forest classifier (RFC), the AUC was [0.897, 95% confidence interval (CI): 0.370-1.424], while the AUC of the artificial neural network, eXtreme gradient boosting, support vector machine, and decision tree were between 0.726 (95%CI: 0.191-1.261) and 0.882 (95%CI: 0.321-1.443). CONCLUSION: Fluctuating serological inflammatory markers and prognostic nutritional index can be used to predict postoperative PF.

Cellular angiofibroma of the vagina: A case report and literature review.

BACKGROUND: Cellular angiofibroma (CAF), a rare benign mesenchymal tumor, is histologically characterized by abundant thick-walled vessels with a spindle cell component. As one of the female reproductive system tumors, its clinical and pathological features are not well characterized. METHODS: A 47-year-old woman presented for the removal of intrauterine device on October 28, 2021, as she had achieved menopause one year back. The patient had no discomfort or awareness of any mass in her vagina. She has history of breast cancer and papillary thyroid cancer. Till date, no progression of thyroid cancer or breast cancer has been observed. Her menstrual cycle was regular, and she had one child delivered vaginally. RESULTS: Pelvic examination revealed a mass sized 2.5 × 2.0 cm located near the fornix in the upper segment of the left vaginal wall. Thin prep cytologic test (TCT) revealed negative intraepithelial lesion or malignancy (NILM). HPV test was negative and leucorrhea routine inspection cleanliness II degree. No cervical mass was detected by ultrasound examination. The patients underwent the operation for intrauterine device removal plus vaginal tumor resection on November 1, 2021. Postoperative antibiotics (intravenous cefuroxime sodium 0.75 g bid for 1 day) were administered to prevent infection. The patient showed no signs of recurrence at one-month follow-up. CONCLUSION: In summary, CAF is a rare benign soft tissue tumor. Surgery is the only treatment method, and the definitive diagnosis of CAF is based on histopathological examination of surgical specimen. Long-term follow-up is needed for surveillance of recurrence.

Survival outcome assessment for triple-negative breast cancer: a nomogram analysis based on integrated clinicopathological, sonographic, and mammographic characteristics.

OBJECTIVE: This study aimed to incorporate clinicopathological, sonographic, and mammographic characteristics to construct and validate a nomogram model for predicting disease-free survival (DFS) in patients with triple-negative breast cancer (TNBC). METHODS: Patients diagnosed with TNBC at our institution between 2011 and 2015 were retrospectively evaluated. A nomogram model was generated based on clinicopathological, sonographic, and mammographic variables that were associated with 1-, 3-, and 5-year DFS determined by multivariate logistic regression analysis in the training set. The nomogram model was validated according to the concordance index (C-index) and calibration curves in the validation set. RESULTS: A total of 636 TNBC patients were enrolled and divided into training cohort (n = 446) and validation cohort (n = 190). Clinical factors including tumor size > 2 cm, axillary dissection, presence of LVI, and sonographic features such as angular/spiculated margins, posterior acoustic shadows, and presence of suspicious lymph nodes on preoperative US showed a tendency towards worse DFS. The multivariate analysis showed that no adjuvant chemotherapy (HR = 6.7, 95% CI: 2.6, 17.5, p < 0.0005), higher axillary tumor burden (HR = 2.7, 95% CI: 1.0, 7.1, p = 0.045), and ≥ 3 malignant features on ultrasound (HR = 2.4, CI: 1.1, 5.0, p = 0.021) were identified as independent prognostic factors associated with poorer DFS outcomes. In the nomogram, the C-index was 0.693 for the training cohort and 0.694 for the validation cohort. The calibration plots also exhibited excellent consistency between the nomogram-predicted and actual survival probabilities in both the training and validation cohorts. CONCLUSIONS: Clinical variables and sonographic features were correlated with the prognosis of TNBCs. The nomogram model based on three variables including no adjuvant chemotherapy, higher axillary tumor load, and more malignant sonographic features showed good predictive performance for poor survival outcomes of TNBC. KEY POINTS: • The absence of adjuvant chemotherapy, heavy axillary tumor load, and malignant-like sonographic features can predict DFS in patients with TNBC. • Mammographic features of TNBC could not predict the survival outcomes of patients with TNBC. • The nomogram integrating clinicopathological and sonographic characteristics is a reliable predictive model for the prognostic outcome of TNBC.

Short-term outcomes of Oxford unicompartmental knee arthroplasty with coronal subluxation of the knee: a retrospective case-control study.

BACKGROUND: The goal of this study was to assess short-term outcomes in single compartment osteoarthritis patients associated with the coronal tibiofemoral subluxation (CTFS) of the knee joint after Oxford unicompartmental knee arthroplasty (OUKA), and to establish the potential impact of the degree of CTFS on operative outcomes. METHODS: Data pertaining to 183 patients with medial compartment osteoarthritis that underwent OUKA treatment between February 2016 and June 2019 were retrospectively analyzed. The presence and degree of severity of CTFS were assessed using preoperative weight-bearing anteroposterior X-ray images of the knee. Patients were stratified into three subgroups based upon the observed degree of subluxation: a normal group, a mild subluxation group (CTFS < 0.5 cm), and a severe subluxation group (CTFS ≥ 0.5 cm). Anterior and posterior X-ray examination of the knee was conducted at the time of most recent follow-up for each patient to assess the degree of CTFS correction following OUKA. Clinical function was assessed using Oxford knee score (OKS) and Hospital for Special Surgery score (HSS) values, while pain was rated using visual-analog scale (VAS) scores. The mechanical femoral tibial angle (mFTA), range of motion (ROM), and complication rates in these three groups were additionally compared. RESULTS: The average follow-up duration for patients in this study was 24.1 months (range: 17-32 months). There were no significant differences in patient age, sex, body mass index (BMI), follow-up duration, mFTA, ROM, OKS, HSS, or VAS scores among these three groups (P > 0.05). After surgery, OKS and HSS scores declined significantly, but no differences in these scores were observed among groups (P > 0.05). Of these patients, 135 (73.8%) were satisfied with the operation, of whom 80 (43.7%) were very satisfied. There were no significant differences in ROM or VAS scores among groups (P > 0.05). The degree of CTFS for patients in the mild and severe subluxation groups was significantly improved following OUKA relative to preoperative values such that the degree of postoperative CTFS did not differ significantly among these groups (P > 0.05). Postoperative mFTA was also significantly improved in these three patient subgroups (P < 0.05). No patients experienced operative complications over the follow-up period. CONCLUSIONS: OUKA can successfully improve clinical symptoms in patients with single compartmental osteoarthritis. Moreover, OUKA can effectively correct CTFS of the knee in these patients, and the degree of preoperative CTFS has no impact on surgical efficacy. LEVEL OF EVIDENCE: III.

Pediatric asthma control during the COVID-19 pandemic: A systematic review and meta-analysis.

BACKGROUND: With the onset of the coronavirus disease 2019 (COVID-19) pandemic, many experts expected that asthma-associated morbidity because of severe acute respiratory syndrome coronavirus 2 infection would dramatically increase. However, some studies suggested that there was no apparent increasing in asthma-related morbidity in children with asthma, it is even possible children may have improved outcomes. To understand the relationship between the COVID-19 pandemic and asthma outcomes, we performed this article. METHODS: We searched PubMed, Embase, and Cochrane Library to find literature from December 2019 to June 2021 related to COVID-19 and children's asthma control, among which results such as abstracts, comments, letters, reviews, and case reports were excluded. The level of asthma control during the COVID-19 pandemic was synthesized and discussed by outcomes of asthma exacerbation, emergency room visit, asthma admission, and childhood asthma control test (c-ACT). RESULTS: A total of 22,159 subjects were included in 10 studies. Random effect model was used to account for the data. Compared with the same period before the COVID-19 pandemic, asthma exacerbation reduced (odds ratio [OR] = 0.26, 95% confidence interval [CI] = [0.14-0.48], Z = 4.32, p < 0.0001), the odds of emergency room visit decreased as well (OR = 0.11, 95% CI = [0.04-0.26], Z = 4.98, p < 0.00001). The outcome of asthma admission showed no significant difference (OR = 0.84, 95% CI = [0.32-2.20], Z = 0.36, p = 0.72). The outcome of c-ACT scores were not analyzed because of the different manifestations used. Overall, c-ACT scores reduced during the pandemic. CONCLUSION: Compared to the same period before the COVID-19 pandemic, the level of asthma control has been significantly improved. We need to understand the exact factors leading to these improvements and find methods to sustain it.

Arthroscopic Internal Drainage with Cyst Wall Resection and Arthroscopic Internal Drainage with Cyst Wall Preservation to Treat Unicameral Popliteal Cysts: A Retrospective Case-Control Study.

OBJECTIVE: To compare the clinical efficacy and safety of arthroscopic internal drainage for the treatment of unicameral popliteal cysts with or without cyst wall resection. METHODS: This was a retrospective case-control study of 73 patients diagnosed with unicameral popliteal cysts from January 2012 to January 2019 who received arthroscopic treatment. The study included 38 cases with cyst wall resection (CWR group) and 35 cases with cyst wall preservation (CWP group). The CWR group consisted of 14 men and 24 women with an average age of 51.8 years, while the CWP group consisted of 13 men and 22 women with an average age of 52.0 years. All patients were examined for intra-articular lesions and communicating ports by magnetic resonance imaging (MRI) prior to surgery, and recurrence of cysts was evaluated at the last follow-up examination. Rauschning and Lindgren grade (R-L grade) and Lysholm score were used to evaluate clinical outcomes. In addition, operation time and complications were recorded. RESULTS: The average length of follow-up was 24.2 months (range, 16 to 32 months). There were no considerable differences in age, gender, cyst size, Lysholm score, R-L grade and concomitant intra-articular cases between the CWR group and CWP group prior to surgery (P > 0.05). The last follow-up MRI scans showed that in the CWR group, the cyst disappeared in 25 cases and shrunk in 13 cases. In the CWP group, the cyst disappeared in 22 cases, shrunk in 12 cases and persisted in one case. There was no obvious difference in recurrence rate between the two groups (0% vs 2.9%, P = 0.899). At the last follow-up, there were no differences in the R-L grade (P = 0.630) and Lysholm score (88.3 ± 5.6 points vs 90.1 ± 3.8 points, P = 0.071) between the two groups. Compared with the CWP group, operation time was significantly prolonged in the CWR group (38.3 ± 3.1 min vs 58.3 ± 4.4 min, P < 0.05). In the CWR group, three cases occurred fluid infiltration under the gastrocnemius muscle, which improved after pressure bandaging and cold compress. In another three cases, hematoma was found. The incidence of complications in the CWR group was markedly higher than that in the CWP group (15.8% vs 0%, P < 0.05). During the follow-up period, none of the patients developed serious complications such as neurovascular injury, deep venous thrombosis, or infection. CONCLUSION: For unicameral popliteal cysts, arthroscopic internal drainage combined with resection of the cyst wall did not further improve the clinical outcomes or reduce the recurrence rate, while prolonging the operation time and increasing the possibility of complications.

Soluplus-Mediated Diosgenin Amorphous Solid Dispersion with High Solubility and High Stability: Development, Characterization and Oral Bioavailability.

BACKGROUND AND PURPOSE: The traditional Chinese medicine, diosgenin (Dio), has attracted increasing attention because it possesses various therapeutic effects, including anti-tumor, anti-infective and anti-allergic properties. However, the commercial application of Dio is limited by its extremely low aqueous solubility and inferior bioavailability in vivo. Soluplus, a novel excipient, has great solubilization and capacity of crystallization inhibition. The purpose of this study was to prepare Soluplus-mediated Dio amorphous solid dispersions (ASDs) to improve its solubility, bioavailability and stability. METHODS: The crystallization inhibition studies were firstly carried out to select excipients using a solvent shift method. According to solubility and dissolution results, the preparation methods and the ratios of drug to excipient were further optimized. The interaction between Dio and Soluplus was characterized by differential scanning calorimetry (DSC), fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), powder X-ray diffraction (PXRD) and molecular docking. The pharmacokinetic study was conducted to explore the potential of Dio ASDs for oral administration. Furthermore, the long-term stability of Dio ASDs was also investigated. RESULTS: Soluplus was preliminarily selected from various excipients because of its potential to improve solubility and stability. The optimized ASDs significantly improved the aqueous solubility of Dio due to its amorphization and the molecular interactions between Dio and Soluplus, as evidenced by dissolution test in vitro, DSC, FT-IR spectroscopy, SEM, PXRD and molecular docking technique. Furthermore, pharmacokinetic studies in rats revealed that the bioavailability of Dio from ASDs was improved about 5 times. In addition, Dio ASDs were stable when stored at 40°C and 75% humidity for 6 months. CONCLUSION: These results indicated that Dio ASDs, with its high solubility, high bioavailability and high stability, would open a promising way in pharmaceutical applications.

One-Stage Repair of Posterior Oblique Ligament Avulsion Fracture Combined with Medial Collateral Ligament Injury.

OBJECTIVE: To evaluate the clinical effect of the one-stage repair of a posterior oblique ligament avulsion fracture combined with a medial collateral ligament injury. METHODS: This study was a retrospective trial. From February 2007 to May 2017, five patients with posterior oblique ligament avulsion fracture combined with medial collateral ligament injury were included in this study. The patients were aged 37-58 years old with a mean of 45.2 years. All patients underwent the primary repair of a posterior oblique ligament avulsion fracture and medial collateral ligament injury. The main observational index included Lysholm score, International Knee Documentation Committee (IKDC) score, Visual Analogue Scale (VAS) score, and range of motion (ROM). RESULTS: The results showed that the average time of follow-up was 53.6 months (range, 20-86 months). When compared to preoperative scores, the preoperative Lysholm score was significantly increased (47.8 ± 5.1 vs 95.0 ± 3.7, P < 0.05), the IKDC score was significantly increased (51.2 ± 5.6 vs 88.6 ± 4.2, P < 0.05), the VAS score was significantly decreased (7.0 ± 0.7 vs 0.4 ± 0.5, P < 0.05), and the ROM was significantly increased (91.6° ± 8.4° vs 129.9° ± 4.4°, P < 0.05). CONCLUSION: Our study found that with the combination of the one-stage repair of a posterior oblique ligament (POL) avulsion fracture and medial collateral ligament injury, the patient's postoperative function recovered well, their pain was relieved, and their knee joint stability was reliable.

Estrogen Stimulation of Pleiotrophin Enhances Osteoblast Differentiation and Maintains Bone Mass in IGFBP-2 Null Mice.

Insulin-like growth factor binding protein-2 (IGFBP-2) stimulates osteoblast differentiation but only male Igfbp2 null mice have a skeletal phenotype. The trophic actions of IGFBP-2 in bone are mediated through its binding to receptor tyrosine phosphatase beta (RPTPß). Another important ligand for RPTPß is pleiotrophin (PTN), which also stimulates osteoblast differentiation. We determined the change in PTN and RPTPß in Igfbp2-/- mice. Analysis of whole bone mRNA in wild-type and knockout mice revealed increased expression of Ptn. Rptpß increased in gene-deleted animals with females having greater expression than males. Knockdown of PTN expression in osteoblasts in vitro inhibited differentiation, and addition of PTN to the incubation medium rescued the response. Estradiol stimulated PTN secretion and PTN knockdown blocked estradiol-stimulated differentiation. PTN addition to IGFBP-2 silenced osteoblast stimulated differentiation, and an anti-fibronectin-3 antibody, which inhibits PTN binding to RPTPß, inhibited this response. Estrogen stimulated PTN secretion and downstream signaling in the IGFBP-2 silenced osteoblasts and these effects were inhibited with anti-fibronectin-3. Administration of estrogen to wild-type and Igfbp2-/- male mice stimulated an increase in both areal bone mineral density and trabecular bone volume fraction but the increase was significantly greater in the Igfbp2-/- animals. Estrogen also stimulated RPTPß expression in the null mice. We conclude that loss of IGFBP-2 expression is accompanied by upregulation of PTN and RPTPß expression in osteoblasts, that the degree of increase is greater in females due to estrogen secretion, and that this compensatory change may account for some component of the maintenance of normal bone mass in female mice.