Synthetic bone grafting with preserved cartilage flap via a medial malleolus osteotomy approach to treat osteochondral lesion of the talus: technical note and preliminary clinical results.

PURPOSE: Although various surgical procedures are available for osteochondral lesion of the talus (OLT), there is still no consensus on its best treatment. The purposes of this study were to describe a new surgical technique to treat OLT and to analyze its preliminary clinical results. METHODS: Eight patients were enrolled in this retrospective study between March 2019 and May 2022 in the Second Affiliated Hospital of Chongqing Medical University. All patients were treated by synthetic bone grafting with preserved cartilage flap via a medial malleolus osteotomy approach. The patients' characteristics, operative time, and estimated blood loss were evaluated. Intraoperative photos, preoperative and postoperative X-ray and MRI imaging were recorded. The American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and visual analog scale (VAS) score were also recorded before surgery and at each follow-up. RESULTS: At six months after the operation, all patients showed bone ingrowth and remodeling according to X-ray and MRI. No obvious defects or ladder was found on the cartilage surface of all patients according to MRI. The AOFAS score improved from 61.63 ± 8.85 (range, 49-74) to 91.13 ± 4.49 (range, 83-97) (p < 0.001) and VAS score improved from 5.50 ± 1.60 (range, 4-8) before surgery to 1.88 ± 0.83 (range, 1-3) (p < 0.001) at latest follow-up. In all eight patients, no wound infection, skin necrosis, or delayed healing of osteotomy was found. CONCLUSION: We proposed a simple and effective technique that restored the shape of the cartilage surface by preserving the cartilage flap and restoring the natural congruency of the subchondral bone by synthetic bone grafting. We found satisfying clinical outcomes in short-term follow-up. Our new technique might be a new surgical option for the treatment of OLT and its effectiveness should be further evaluated.

The Postoperative Platelet to Creatinine Ratio as A Prognostic Index of In-Hospital Mortality in Patients with Acute Type A Aortic Dissection.

BACKGROUND: The purpose of the investigation was to assess the value of post-operation platelet to creatinine ratio (PCR) in predicting in-hospital mortality among patients with acute type A aortic dissection (TAAAD). METHODS: A retrospective study was carried out from January 2017 to December 2019. The best cutoff value of post-operation PCR was assessed by receiver operating characteristic (ROC) curve. Patients were divided into survivors and nonsurvivors. Univariate and multivariate logistic analyses were carried out to identify independent risk factors influencing in-hospital mortality. RESULTS: A total of 171 patients were included in this investigation, with an in-hospital mortality rate of 18.1%. The optimal cut-off value of post-operation PCR was 0.7242 (area under the ROC curve (AUC): 0.798, 95% confidence interval (CI) 0.730-0.856, p < 0.001), and the sensitivity and specificity were 74.2% and 74.3%. The levels of post-operation PCR were lower in nonsurvivors than in survivors (0.56 ± 0.33 vs. 1.50 ± 1.36, p < 0.001). Multivariate logistic regression analysis displayed that post-operation PCR was positively related to in-hospital survivors when confounding factors were adjusted (HR = 8.850, 95% CI = 2.611-30.303, p < 0.001). CONCLUSIONS: Post-operative PCR is a readily accessible and cost-effective biomarker that is independently associated with in-hospital mortality in TAAAD patients. Furthermore, it exhibits superior performance in predicting patient outcomes following surgery.

The articular cartilage preservative effects of genistein in an experimental model of knees osteoarthritis.

The study aimed to investigate the preservative effects of genistein on articular cartilage in an experimental model of knee osteoarthritis in rats. Thirty male Wistar rats were assigned to 3 equal groups: sham group, osteoarthritis control group (OAG), and genistein-treated osteoarthritis group (GTG). Intra-articular injections of monosodium iodoacetate were used for osteoarthritis induction. After 2 weeks of rest for the induction of the inflammatory process, genistein (30 mg/kg/day) vs. saline gavage was administered for 8 weeks. The expression of matrix metalloproteinase (MMP)-8 and MMP-13, Sox5/Sox6, Indian hedgehog (IHH), and Col2 were evaluated in medial femoral condyle sections by immunohistochemical staining. The number of chondrocytes and cartilage thicknesses were also measured and compared among the groups. No significant change in cartilage thickness was observed in GTG compared with OAG (p = 0.188). Chondrocyte count was significantly higher in the articular cartilage of GTG compared with OAG (p = 0.006). Induction of osteoarthritis significantly increased the expression of MMP-8, MMP-13, and IHH, but decreased Col2, Sox5, and Sox6 expression (p < 0.001); these were partially prevented in the GTG. Our findings support the effectiveness of genistein treatment in the prevention of articular cartilage damage in the experimental model of knee osteoarthritis. The proposed mechanism of action is through the suppression of the MMP, IHH, and Col2 pathways, besides the induction of Sox5 and Sox6 expression. Novelty: Genistein prevents articular cartilage damage in the experimental model of knee osteoarthritis. The osteoprotective effect is manifested by the modulation of expression of MMP, Sox, IHH, and Col2 proteins.

Changes of lipid profiles in human umbilical vein endothelial cells exposed to zirconia nanoparticles with or without the presence of free fatty acids.

Zirconia nanoparticles (NPs) have been widely used in biomedicine, which will likely lead to their interactions with endothelial cells (ECs). However, the toxicity of zirconia NPs to ECs is less investigated and the toxicological data are not consistent. Furthermore, no previous study, to the best of our knowledge, investigated the influence of zirconia NPs on lipid metabolism. This study investigated lipid profiles in human umbilical vein ECs (HUVECs) exposed to zirconia NPs with or without the presence of free fatty acids (FFAs). Incubation with FFA changed the hydrodynamic size, zeta potential, and surface profiles of zirconia NPs, indicating the surface coating effects. Exposure of HUVECs to various concentrations of zirconia NPs with or without the presence of FFA did not significantly decrease cellular viability, but FFA decreased zirconium elemental levels in NP-exposed cells. Oil Red O staining showed that FFA or zirconia NPs and FFA, but not zirconia NPs alone, significantly increased lipid accumulation in HUVECs. Consistently, lipidomic data suggested that exposure to FFA or zirconia NPs and FFA up-regulated most lipid classes in HUVECs. As the mechanisms for increased lipid accumulation, exposure to FFA or zirconia NPs and FFA up-regulated endoplasmic reticulum (ER) stress axis IRE1α-XBP-1, leading to increased FASN and ACSL3, proteins involved in lipid metabolism. Combined, our results demonstrated that zirconia NPs were noncytotoxic and showed minimal impact on ER stress-mediated lipid metabolism in HUVECs under both normal and FFA-challenged conditions, which indicated the relatively high biocompatibility of zirconia NPs to ECs.

GATA1/SP1 and miR-874 mediate enterovirus-71-induced apoptosis in a granzyme-B-dependent manner in Jurkat cells.

Enterovirus 71 (EV71)-induced T lymphocyte apoptosis plays an important role in hand, foot, and mouth disease (HFMD), and granzyme B (GZMB) has been shown to be critical for this process. However, the mechanisms underlying GZMB-mediated apoptosis of T lymphocytes remain unknown. In this study, we investigated whether transcription factors and microRNAs (miRNAs) are involved in GZMB-mediated apoptosis of T lymphocytes in response to EV71 infection. Our findings indicated that EV71 infection significantly induced apoptosis in Jurkat cells, a human T lymphocytes cell line, as revealed in flow cytometric analysis. Furthermore, EV71 increased the expression of pro-apoptosis Bcl-2-associated X (Bax) and cleaved caspase 3 but decreased the expression of anti-apoptosis B-cell lymphoma protein 2 (Bcl2). GZMB knockdown decreased cell apoptosis and prevented EV71-induced changes in the expression of Bax, cleaved caspase 3, and Bcl2 in Jurkat cells, highlighting the role of GZMB as a key factor in EV71-induced apoptosis. Our study also indicated that overexpression of the transcription factors GATA binding factor 1 (GATA1) and specificity protein 1 (SP1) significantly increased luciferase activity when this gene was inserted in the GZMB 3' untranslated region (3'UTR). GATA1/SP1 overexpression induced cell apoptosis, increased the expression of Bax and cleaved caspase 3, and decreased the expression of Bcl2. Finally, our results suggested that miR-874 plays an essential role in GZMB-mediated cell apoptosis, since an miR-874 mimic decreases the expression of GZMB by targeting its 3'UTR. Collectively, these data indicated that GATA1/SP1 and miR-874 mediate EV71-induced apoptosis in a granzyme B-dependent manner. This signaling pathway may provide a new pharmacological target for the prevention and treatment of HFMD.

Wetlands of International Importance: Status, Threats, and Future Protection.

The 2303 Wetlands of International Importance distribute unevenly in different continents. Europe owns the largest number of sites, while Africa has the largest area of sites. More than half of the sites are affected by three or four impact factors (55%). The most significant impact factors are pollution (54%), biological resources use (53%), natural system modification (53%), and agriculture and aquaculture (42%). The main affected objects are land area and environment of the wetlands, occurred in 75% and 69% of the sites, respectively. The types most affected by land area occupation are river wetlands and lake wetlands, the types with the greatest impact on environment are marine/coastal wetlands and river wetlands, the type with the greatest impact on biodiversity is river wetlands, the types most affected by water resources regulation are marsh wetlands and river wetlands, and the types most affected by climate change are lake wetlands and marine/coastal wetlands. About one-third of the wetland sites have been artificially reconstructed. However, it is found that the proportions of natural wetland sites not affected or affected by only one factor are generally higher than that of wetland sites both containing natural wetlands and human-made wetlands, while the proportions of wetland sites both containing natural wetlands and human-made wetlands affected by three or four factors are generally higher than that of natural wetland sites. Wetland sites in the UK and Ireland are least affected among all countries. Wetland management plans in different regions still have large space for improvement, especially in Africa and Asia. The protection and restoration of global wetlands can be carried out in five aspects, including management and policy, monitoring, restoration, knowledge, and funding.

Comparative proteomic analysis of plasma of children with congenital heart disease.

Congenital heart disease is one of the largest class of birth defects. Eight subjects with ventricular septal defect (VSD, a kind of congenital heart disease) and 11 health children were enrolled in tandem mass tags label-based quantitative proteomic analysis to compare plasma proteins differentially abundance. A total of 66 proteins were significantly upregulated or downregulated in VSD patients compared with healthy children. These proteins were involved in pathways linked to platelet activation, fructose and mannose metabolism, complement and coagulation cascades, glycolysis/gluconeogenesis, regulation of actin cytoskeleton, and carbon metabolism. The amount of ten proteins changed significantly (p < 0.05) in newly recruited 30 VSD compared with 15 control children, which were validated by ELISA. The areas under the receiver operating characteristic curve values of fructose-bisphosphate aldolase B (ALDOB) and thymosin beta-4 (Tß4) were higher than those of other candidate proteins. ALDOB and Tß4 might be potential biomarkers applied for identifying VSD in the further works.

Characteristics of NDM-1-producing Klebsiella pneumoniae ST234 and ST1412 isolates spread in a neonatal unit.

BACKGROUND: The emergence of carbapenem-resistant Klebsiella pneumoniae (CR-KP) has become a significant problem worldwide and also being a major threat to children and newborns. Here we report an outbreak of NDM-1-producing K. pneumoniae in a neonatal unit. RESULTS: Six CR-KP strains, isolated from neonates with symptoms of infection, were identified using a VITEK-2 compact system, and the clinical data were retrieved from the electronic case records. In vitro susceptibility testing with broth dilution method showed that all six K. pneumoniae isolates were resistant to carbapenems and susceptible to colistin, aminoglycosides, fluoroquinolones and tigecycline. Based on the polymerase chain reaction results, each isolate was found to be blaNDM-1 gene positive. Clonal relationships were analysed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and showed that two different PFGE patterns were formed, which belonged to sequence types ST234 and ST1412. Plasmids carrying blaNDM-1 were successfully transferred from four of the six isolates to an Escherichia coli recipient through conjugative assays. S1-PFGE and Southern blot hybridization showed that four NDM-1-producing K. pneumoniae were clonal and carried blaNDM-1 on the same plasmid. The outbreak was effectively controlled by reducing the potential infection sources. All the patients were successfully treated and recovered after receiving an increased dose of carbapenems. Although the source of this outbreak was not clear, comprehensive measures were carried out and the outbreak was effectively controlled. CONCLUSIONS: ST234 and ST1412 of NDM-1-producing Klebsiella pneumoniae are the resistant clone spread in the neonatal unit, comprehensive infection control measures and optimized carbapenem therapy played an important role in controlling this NDM-1-producing K. pneumoniae outbreak.

Serum SP70 is a sensitive predictor of chemotherapy response in patients with advanced nonsmall cell lung cancer.

SP70 is a novel tumor biomarker in patients with nonsmall cell lung cancer (NSCLC). However, its role as a marker for predicting the response to chemotherapy for patients with advanced NSCLC has not been investigated. A total of 152 patients were enrolled. Serum SP70, carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1), and neuron-specific enolase (NSE) were detected before and after 2 cycles of chemotherapy. The correlation between serum tumor biomarker levels and chemotherapy responses and their association with epidermal growth factor receptor (EGFR) mutation status and progression-free survival (PFS) were analyzed. Serum SP70 levels were significantly decreased after chemotherapy in the partial remission (PR) group (P < .001) and increased in the progressive disease (PD) group (P < .001), but not significantly changed in the stable disease (SD) group (P = .114). Although similar changes were observed on CEA and CYFRA21-1 levels but not NSE, ROC analysis demonstrated that SP70 is superior to the others. Additionally, patients with EGFR mutation had higher serum SP70 levels and tissue SP70 expression than patients without EGFR mutation (P = .014 and P = .002, respectively). The median PFS of patients with decreased SP70 levels after chemotherapy was longer than that of patients with stable or increased serum SP70 level (24 months vs 12 months vs 2 months, P < .001), and the differences of all other 3 tumor markers were not obvious. Serum SP70 is a sensitive and real-time indicator of chemotherapeutic efficacy in patients with advanced NSCLC and related to PFS.

Treatment of acute achilles tendon rupture with the panda rope bridge technique.

INTRODUCTION: Although nonsurgical methods and many surgical techniques have been developed for repairing a ruptured Achilles tendon, there is no consensus on its best treatment. In this article, a novel minimally invasive technique called the Panda Rope Bridge Technique (PRBT) is described. METHODS: Patient with acute Achilles tendon rupture was operated on in the prone position. The PRBT begin with making the proximal bridge anchor (Krackow sutures in the myotendinous junction), the distal bridge anchor (two suture anchors in the calcaneus bone) and the ropes (threads of the suture anchors) stretched between the anchor sites. Then a small incision was made to debride and reattach the stumps of ruptured tendon. After the surgery, no cast or splint fixation was applied. All patients performed enhanced recovery after surgery (ERAS), which included immediate ankle mobilisation from day 1, full weight-bearing walking from day 5 to 7, and gradually take part in athletic exercises from 8 weeks postoperatively. RESULTS: PBRT was performed in 11patients with acute Achilles tendon rupture between June 2012 and June 2015. No wound infection, fistula, skin necrosis, sural nerve damage, deep venous thrombosis or tendon re-rupture was found. One year after the surgery, all patients reported 100 AOFAS ankle-hindfoot score points and the mean ATRS was 96.6. CONCLUSION: The PRBT is a simple, effective and minimally invasive technique, with no need for immobilisation of the ankle, making possible immediate and aggressive postoperative rehabilitation.

Knee joint changes in patients with neglected developmental hip dysplasia: a prospective case-control study.

BACKGROUND: Few reports are available describing knee changes in neglected developmental dysplasia of the hip (DDH). The purpose of this study was to assess the radiographic morphology of knee joints in adults with neglected DDH. METHODS: Thirty-seven patients (35 females and two males) with neglected DDH were prospectively recruited with an average age of 32.6 years. Twenty-three patients had unilateral and 14 patients had bilateral neglected DDH. Thirty-seven healthy individuals were recruited to form a matched control group. Three groups of knee joints were examined: affected knees (on the same side of the neglected DDH), unaffected knees (contralateral to the neglected DDH in patients with unilateral involvement), and control knees. A series of radiographic parameters of the knee joint were measured in the coronal and sagittal plane, and they were compared between patients and normal controls. RESULTS: In the coronal plane, the affected knees had increased valgus angulation related to increased height of the medial femoral condyle, decreased height of the lateral femoral condyle and decreased lateral distal femoral angle compared to control knees. In the sagittal plane, both distal femoral and proximal tibial joints of the affected knees developed a decrease in posterior angles. Additionally, the unaffected knees also developed radiographic changes compared to control knees. CONCLUSIONS: Patients with neglected DDH may develop changes in both knee joints. These changes should be considered during surgery to the hip, femur and knee to prevent potential complications. LEVEL OF EVIDENCE: Level 2.

Heritability and sibling recurrent risk of developmental dysplasia of the hip in Chinese population.

BACKGROUND: Previous familial segregation studies supported that developmental dysplasia of the hip (DDH) is a multifactorial genetic disease. However, the exact extent of genetic effects has not been fully evaluated, especially in Asian population. The aim of this study is to estimate the sibling recurrent risk and heritability of DDH in a large Chinese cohort. MATERIALS AND METHODS: Four hundred and twenty-nine DDH probands and 534 matched normal controls were recruited from a screening programme for DDH, including 628 siblings in families of probands and 889 siblings in those of controls, respectively. The detailed information of family history was obtained, and the prevalence of DDH among siblings of probands, as well as controls, was determined. The sibling recurrent risk and heritability was estimated using classical liability threshold model. RESULTS: Eighty-seven siblings (13.85%) in families of proband and 14 siblings (1.57%) in normal control families were diagnosed as DDH. The recurrent risk in siblings of probands was at least tenfolds that in siblings of controls. Compared with the normal controls, the sibling recurrent risk was about 12-fold increase in male sib, and 9-fold increase in female sib. Overall, a high heritability of 83.59 ± 4.90% (h(2) ± SE) was observed. When stratified by genders, it was even higher for female siblings (91.02 ± 7.25%). CONCLUSION: This study for the first time exhibits a high sibling recurrent risk and heritability for DDH in Asian population. It also shows there is a high probability to identify the underlying predisposition genes in future genetic study.

Overexpression of GDF5 through an adenovirus vector stimulates osteogenesis of human mesenchymal stem cells in vitro and in vivo.

The use of stem cells combined with gene therapy could be an important way to facilitate bone regeneration. In this study, the aim was to investigate the potential of growth and differentiation factor-5 (GDF5) to genetically manipulate human mesenchymal stem cells (hMSCs) for bone regeneration. Recombinant adenovirus Ad-GDF5 and Ad-GFP were constructed and identified, and the titer of both were determined. Third-passage hMSCs were infected with adenovirus, and the expression of GDF5 was confirmed by detection of GFP-positive cells, GDF5 mRNA levels, Western blotting, and enzyme-linked immunosorbent assay (ELISA). hMSCs at passage 3 were divided into four groups: (1) an experimental group infected with Ad-GDF5, (2) a positive control group cultured with osteogenic differentiation medium, (3) a control group infected with Ad-GFP cultured with standard medium, and (4) a blank control group cultured with standard medium. Evaluation of cell morphology and proliferation, analysis of the expression of genes related to osteogenic differentiation, von Kossa staining, and immunofluorescent staining of collagen I were used to investigate the osteogenesis of cells among the groups. After culturing the cells for 2 days under each corresponding condition, the cells were detached and subcutaneously injected into the backs of nude mice to evaluate bone formation. Samples were collected for histological staining, protein Western blotting, and micro-computer tomography. When infected with Ad-GDF5, hMSCs could overexpress GDF5 for a prolonged period in vitro and reach a concentration of 160 ng/ml. Cells infected with Ad-GDF5 or cultured in osteogenic medium displayed osteogenic differentiation based on their histological and cellular properties and on their gene and protein expression patterns. Furthermore, Ad-GDF5 showed a better ability to upregulate the expression of collagen I, alkaline phosphatase, and osteocalcin mRNA than the osteogenic medium. Furthermore, Ad-GDF5 expression was associated with enhanced bone formation in vivo. Our findings suggest that hMSCs infected with Ad-GDF5 can differentiate in an osteogenic direction and may be a promising cell source for bone regeneration.

[Construction of human GDF-5 on adenovirus vector and its expression in MSCs].

This experimental study was aimed to construct the recombinant adenovirus vector containing human GDF-5 gene, and to use it for infecting human MSCs and detecting the expression of the gene GDF-5. The core sequence of human GDF-5 was amplified by PCR from pCMV-SPORT6, and then was cloned to pAdtrack-CMV. The linearized shuttle plasmid pAdtrack-CMV-GDF-5 was homogenously recombined with pAdeasy-1 in BJ5183. The potential clone was analyzed by restriction endonuclease digestion. The correct clone was linearized and transfected into QBI-293 cells for packing and amplifying so as to obtain adenovirus pAd-GDF-5 and identify it, while the titer was also determined by TCID50. MSCs were infected by the harvested virus, and the expression of GDF-5 was detected by RT-PCR. The recombinant adenovirus vector containing human GDF-5 gene was constructed successfully, its titer was 1 x 10(9) PFU/ml, and it could infect MSCs efficiently. The human MSCs infected by constructed adenovirus vector could continue expressing GDF-5 in a certain time, and the transgenic MSCs would be much potential on tissue regeneration.

[Effect of growth and differentiation factor 5 gene transferred by adenovirus on hBMSCs osteogenic differentiation].

OBJECTIVE: To introduce growth and differentiation factor 5 (GDF-5) gene into hBMSCs using recombinant adenovirus vector and to investigate the effect of GDF-5 gene expression on hBMSCs osteogenic differentiation. METHODS: Recombinant adenovirus GDF-5 (Ad-GDF-5) containing green fluorescent protein (GFP) and Ad-GFP were amplified and titered. hBMSCs at passage 3 were infected with two viruses at different titers. At 2 days after intervention, GFP expression was observed using fluorescence microscope, and GDF-5 expression in hBMSCs was detected by RT-PCR. Adherent hBMSCs at passage 3 were randomly divided into 4 groups: experimental group (GDF-5 gene transfection), osteogenic induction group, Ad-GFP infection group, and control group. Cell differentiation was detected by inverted phase contrast microscope observation, fluorescence microscope observation, reverse transcription fluorescence quantitative PCR, immunofluorescence staining, and von Kossa staining at different time points after intervention. RESULTS: The titer of Ad-GDF-5 and Ad-GFP was 1.0 x 10(9) pfu/mL and 1.2 x 10(9) pfu/mL, respectively, hBMSCs was efficiently infected by Ad-GDF-5 and Ad-GFP, and expressed target gene and GFP gene. At 1-7 days after intervention, morphology and growth pattern of the hBMSCs in the experimental group and the osteogenic induction group were transformed into osteoblast-like cells, whereas the cells in the other two groups were still maintained their original morphology and growth pattern. Reverse transcription fluorescence quantitative PCR detection: at 4 days after intervention, GDF-5 expression in the experimental group was obviously higher than that of other groups (P < 0.05); ALP, Col I, and OC gene expression in the experimental and the osteogenic induction group were superior to those of the Ad-GFP infection and the control group (P < 0.05); Col I gene expression in the osteogenic induction group was greater than that of the experimental group (P < 0.05). Immunofluorescence staining: at 4 days after intervention, the cells in the osteogenic induction group and the experimental group expressed and secreted Col I, and no expression of Col I was evident in the other two groups. At 10 days after intervention, the cells in the osteogenic induction and the experimental group were positive for von Kossa staining, and the results of the other two groups were negative. CONCLUSION: GDF-5 gene can be transferred into hBMSCs via adenovirus vector and be expressed stably. It can facilitate the osteogenic differentiation of the hBMSCs and lay a foundation for the further study of this kind of gene transferred hBMSCs effect on bone tissue repair.