A method for extraction of exosomes from breast tumour cells and characterisation by transmission electron microscopy.

Exosomes can not only be used as markers of tumour metastasis but also be used for noninvasive diagnosis of clinical diseases, which holds significant medical research value. Observing the morphology and distribution of exosomes using transmission electron microscopy (TEM) is highly significant. In this study, we use breast tumour cell (MDA-MB-231) exosomes as a representative sample and focus on the extraction and purification of exosomes, as well as the investigation of optimal conditions for the observation of exosomes using TEM. Through comparative tests, we obtained the optimal dilution concentration and staining method for TEM of exosomes, the best dilution concentration is 100 times, the best negative staining time is 1.5 min. Western blotting and scanning electron microscopy (SEM) confirmed the presence of exosome. The particle size experiment shows that the size of exosomes is mainly distributed between 60 nm and 150 nm. This method provides a reference for TEM sample preparation of subcellular structures and small molecular biomaterials other than exosomes.

Bisphosphonate-incorporated coatings for orthopedic implants functionalization.

Bisphosphonates (BPs), the stable analogs of pyrophosphate, are well-known inhibitors of osteoclastogenesis to prevent osteoporotic bone loss and improve implant osseointegration in patients suffering from osteoporosis. Compared to systemic administration, BPs-incorporated coatings enable the direct delivery of BPs to the local area, which will precisely enhance osseointegration and bone repair without the systemic side effects. However, an elaborate and comprehensive review of BP coatings of implants is lacking. Herein, the cellular level (e.g., osteoclasts, osteocytes, osteoblasts, osteoclast precursors, and bone mesenchymal stem cells) and molecular biological regulatory mechanism of BPs in regulating bone homeostasis are overviewed systematically. Moreover, the currently available methods (e.g., chemical reaction, porous carriers, and organic material films) of BP coatings construction are outlined and summarized in detail. As one of the key directions, the latest advances of BP-coated implants to enhance bone repair and osseointegration in basic experiments and clinical trials are presented and critically evaluated. Finally, the challenges and prospects of BP coatings are also purposed, and it will open a new chapter in clinical translation for BP-coated implants.

Development and Application of nanoPCR Method for Detection of Feline Panleukopenia Virus.

Feline panleukopenia (FP) is a severe viral illness caused by the feline panleukopenia virus (FPV), putting sectors like companion cat breeding and endangered feline conservation at risk. The virus has a high morbidity and fatality rate and is found all over the world. We created a novel FPV assay using nanoPCR technology and assessed the method's specificity and sensitivity. The approach amplified a 345 bp nucleic acid fragment with a minimum detection limit of 7.97 × 102 copies/µL, which is about 100 times greater than traditional PCR. We collected anal swabs from 83 cats suspected of FPV infection for practical application, and the FPV-positive rate determined by the nanoPCR approach was 77.1%. In conclusion, the approach is more sensitive than conventional PCR and more convenient and cost-effective than qPCR methodology and may be utilized for the clinical detection of FPV.

Isolation of feline panleukopenia virus from Yanji of China and molecular epidemiology from 2021 to 2022.

BACKGROUND: Feline panleukopenia virus (FPV) is a widespread and highly infectious pathogen in cats with a high mortality rate. Although Yanji has a developed cat breeding industry, the variation of FPV locally is still unclear. OBJECTIVES: This study aimed to isolate and investigate the epidemiology of FPV in Yanji between 2021 and 2022. METHODS: A strain of FPV was isolated from F81 cells. Cats suspected of FPV infection (n = 80) between 2021 and 2022 from Yanji were enrolled in this study. The capsid protein 2 (VP2) of FPV was amplified. It was cloned into the pMD-19T vector and transformed into a competent Escherichia coli strain. The positive colonies were analyzed via VP2 Sanger sequencing. A phylogenetic analysis based on a VP2 coding sequence was performed to identify the genetic relationships between the strains. RESULTS: An FPV strain named YBYJ-1 was successfully isolated. The virus diameter was approximately 20-24 nm, 50% tissue culture infectious dose = 1 × 10-4.94/mL, which caused cytopathic effect in F81 cells. The epidemiological survey from 2021 to 2022 showed that 27 of the 80 samples were FPV-positive. Additionally, three strains positive for CPV-2c were unexpectedly found. Phylogenetic analysis showed that most of the 27 FPV strains belonged to the same group, and no mutations were found in the critical amino acids. CONCLUSIONS: A local FPV strain named YBYJ-1 was successfully isolated. There was no critical mutation in FPV in Yanji, but some cases with CPV-2c infected cats were identified.

The influence of body weight index on initial stability of uncemented femoral knee protheses: A finite element study.

Background and objective: Obesity is one of the risk factors for osteoarthritis. The end-stage treatment for osteoarthritis is total knee arthroplasty (TKA). However, it remains controversial whether a high body mass index (BMI) affects the initial stability of the femoral prosthesis after TKA. Finite element analysis (FEA) was used to investigate this question in this study. Methods: Four femur models that assembled with TKA femoral components were reconstructed and divided into high BMI group and normal BMI group. The three-dimensional femurs were modeled and assigned inhomogeneous materials based on computed tomography (CT) images. Then each FEA model was applied with gait and deep bend loading conditions to evaluate the maximum principal strain on the distal femur and the relative micromotion between the femur and prosthesis. Results: The mean strain of the high BMI group increased by 32.7% (936.9 µÎµ versus 706.1 µÎµ) and 50.9% (2064.5 µÎµ versus 1368.2 µÎµ) under gait and deep bend loading conditions, respectively, compared to the normal BMI group. Meanwhile, the mean micromotion of the high BMI group increased by 41.6% (2.77 µm versus 1.96 µm) and 58.5% (62.1 µm versus 39.2 µm), respectively. Under gait condition, the maximum micromotion for high BMI group was 33.8 µm and would compromise the initial stability. Under deep bend condition, the maximum strain and micromotion exceeded -7300 µÎµ and 28 µm for both groups. Conclusion: High BMI caused higher strain on the bone and higher micromotion between the prosthesis and the femur. Gait activities could be risky for prosthesis stability in high BMI group while be safe in normal group. Deep bend activities were highly dangerous for both groups with high BMI and normal BMI and should be avoided.

Comparison of the relative efficacy of ß-lactam/ß-lactamase inhibitors and carbapenems in the treatment of complicated urinary tract infections caused by ceftriaxone-non-susceptible Enterobacterales: a multicentre retrospective observational cohort study.

BACKGROUND: Treating complicated urinary tract infections (cUTIs) caused by ESBL-producing Enterobacterales represents a significant clinical challenge. The present study was thus developed to explore the relative efficacy of ß-lactam/ß-lactamase inhibitors (BLBLIs) and carbapenems for the treatment of hospitalized patients suffering from cUTIs caused by BLBLI-susceptible ceftriaxone-non-susceptible Enterobacterales. METHODS: Data from 557 patients from four Chinese teaching hospitals diagnosed with cUTIs caused by ceftriaxone-non-susceptible Enterobacterales from January 2017 to May 2022 were retrospectively assessed. RESULT: The 30 day rate of treatment failure, defined by unresolved symptoms or mortality, was 10.4% (58/557). Independent predictors of 30 day treatment failure included immunocompromised status, bacteraemia, septic shock, lack of infection source control and appropriate empirical treatment. When data were controlled for potential confounding variables, BLBLI treatment exhibited a comparable risk of 14 day (OR 1.61, 95% CI 0.86-3.00, P = 0.133) and 30 day treatment failure (OR 1.45, 95% CI 0.66-3.15, P = 0.354) relative to carbapenem treatment for the overall cohort of patients. In contrast, BLBLI treatment in immunocompromised patients was associated with an elevated risk of both 14 day (OR 3.18, 95% CI 1.43-7.10, P = 0.005) and 30 day treatment failure (OR 3.06, 95% CI 1.07-8.80, P = 0.038) relative to carbapenem treatment. CONCLUSIONS: These results suggested that carbapenem treatment may be superior to BLBLI treatment for immunocompromised patients suffering from cUTIs caused by ceftriaxone-non-susceptible Enterobacterales species. However, these results will need to be validated in appropriately constructed randomized controlled trials to ensure appropriate patient treatment.

Development and Evaluation of NanoPCR for the Detection of Goose Parvovirus.

Gosling plague (GP) is an acute and hemorrhagic infectious disease caused by goose parvovirus (GPV). The goose industry suffers significant economic losses as a result of GP, which is found to be widespread worldwide, with high rates of morbidity and mortality. Our group developed a novel technique for detecting GPV nanoparticle-assisted polymerase chain reaction (nanoPCR) and the characterization of its specificity and sensitivity. It was developed by using the traditional polymerase chain reaction (PCR) and nanoparticles. The findings of this study revealed that GPV nanoPCR products were 389 bp in length, and the lower limit of the nanoPCR assay was 4.68 × 102 copies/µL, whereas that of the conventional PCR assay was 4.68 × 104 copies/µL. A total of 230 geese suspected of GPV were detected using nanoPCR, with a positive rate of 83.0% and a specificity of 73%, respectively. Overall, we present a hitherto undocumented method for identifying GPV by using nanoPCR to aid in the evaluation of subclinical illness.

Molecular epidemiology of canine parvovirus 2 from 2014, 2019, and 2021 shows CPV2 circulating and CPV2c increasing in Yanbian, China.

Canine parvovirus 2 (CPV2) causes one of the most serious canine viral infections, with high mortality in young dogs. In 2014, 2019, and 2021, we determined genetic sequences of CPV2 strains obtained from 39 fecal samples collected from the Yanbian Korean Autonomous Prefecture in the Jilin Province of China. Sequence alignments were performed using the major capsid protein (VP2) gene; protein sequences of these samples had high nucleotide (>97.4%) and amino acid (>95.6%) identity. All of the amino acid sequences contained Ser297Ala and Tyr324Ile mutations. Our survey indicated a high prevalence of CPV2 variants in Yanbian Prefecture, with the new CPV2a variant (26 of 39; 67%) being the most frequent. CPV2c, identified in 9 of 39 (23%) samples, had not been detected in this region previously, indicating the potential risk of CPV2 mutation. The sequences of our 39 CPV2 samples were more highly homologous to the published Chinese strains than to the CPV2 variant strains found in other countries.

Novel Design of the Compound Sleeve and Stem Prosthesis for Treatment of Proximal Femur Bone Defects Based on Topology Optimization.

The loosening of traditional prosthetics is among the leading causes of surgical failure of proximal femoral bone defects. A novel compound sleeve and stem prosthesis was designed using an optimization methodology that combined an octet-truss porous structure with density-based topology optimization to improve stability, promote bone ingrowth, and enhance biomechanical properties. Biomechanical changes were assessed using finite element analysis. The distribution of stress, the strain energy density, and the relative micromotion in the optimized group were considered. The optimized sleeve prosthesis achieved a 31.5% weight reduction. The maximum stresses in the optimized group were observed to decrease by 30.33 and 4.74% at the back sleeve and neck part of stem prosthesis, with a 29.52% increase in the femur, respectively. The average stress in most selected regions in the optimized group was significantly greater than that in the original group (p < 0.05). The maximum relative micromotion decreased by 15.18% (from 63.9 to 54.2 µm) in the optimized group. The novel designed compound sleeve and stem prosthesis could effectively improve the biomechanical performance of next-generation prosthetics and provide a microenvironment for bone ingrowth. The presented method could serve as a model for clinical practice and a platform for future orthopedic surgery applications.

Improving the Stability of a Hemipelvic Prosthesis Based on Bone Mineral Density Screw Channel and Prosthesis Optimization Design.

In pelvic reconstruction surgery, the hemipelvic prosthesis can cause significant changes in stress distribution due to its high stiffness, and its solid structure is not suitable for osseointegration. The purpose of this study was to identify a novel bone mineral density screw channel and design the structure of the prosthesis so as to improve the distribution of stress, promote bone growth, and enhance the biomechanical properties of the prosthesis. The mechanical characteristics of bone mineral density screw and traditional screw were compared by finite element analysis method, and redesigned by topology optimization. The direction of the newly proposed screw channel was the posterolateral entrance of the auricular surface, ending at the contralateral sacral cape. Compared to the original group, the maximum stress of the optimized prosthesis was decreased by 24.39%, the maximum stress of the sacrum in the optimized group was decreased by 27.23%, and the average strain energy density of the sacrum in the optimized group was increased by 8.43%. On the surface of screw and connecting plate, the area with micromotion more than 28 µm is reduced by 12.17%. On the screw surface, the area with micromotion more than 28 µm is reduced by 22.9%. The newly determined screw channel and optimized prosthesis design can effectively improve the biomechanical properties of a prosthesis and the microenvironment of osseointegration. This method can provide a reference for the fixation of prostheses in clinical pelvic reconstruction.

Experimental Study on Microstructure and Erosion Mechanisms of Solid Waste Cemented Paste Backfill under the Combined Action of Dry-Wet Cycles and Sulphate Erosion.

Solid waste cemented paste backfill (SWCPB) meets the needs of coal mining area management. SWCPB is a cementitious paste backfill material without added cement and is made only from oil shale residue (OSR), steel slag (SS), soda residue (SR) and water. In this study, mine water characteristics were simulated by combining dry-wet cycling experiments with sulphate erosion experiments. SWCPB was assessed regarding appearance, mass loss, and unconfined compressive strength (UCS), and the erosion products were microscopically analysed with X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The mechanism for erosion of the SWCPB by sulphate-rich mine water was comprehensively analysed and revealed. Research showed that the erosion mechanism was divided into two parts: chemical and physical erosion. Low concentrations of sodium sulphate promoted hydration, thereby contributing to the increased mass and strength of SWCPB. At high sodium sulphate concentrations, the erosion mainly consumed Ca(OH)2 within the material, and the main generated erosion products were gypsum and ettringite (AFt). This was accompanied by the destructive effects of Na2SO4 crystal expansion, which resulted in damage and the reduced workability of the SWCPB. The whole erosion process was continuous, mainly due to transformations of pits, pores and cracks. The conclusions of this study may provide appropriate guidance for application of SWCPB materials in the treatment of coal mine backfills. In addition, the corresponding theoretical analysis of the erosion mechanism for SWCPB materials is provided.

Preparation of Cemented Oil Shale Residue-Steel Slag-Ground Granulated Blast Furnace Slag Backfill and Its Environmental Impact.

A new environmentally friendly cemented oil shale residue-steel slag-ground granulated blast furnace slag backfill (COSGB) was prepared using oil shale residue (OSR), steel slag (SS) and ground granulated blast furnace slag (GGBS) as constituent materials. Based on univariate analysis and the Box-Behnken design (BBD) response surface method, the three responses of the 28 days unconfined compressive strength (UCS), slump and cost were used to optimize the mix ratio. Using a combination of scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and mercury intrusion porosimetry (MIP), the reaction products, microscopic morphology and pore structure of the specimens with the optimal mix ratio at different curing ages were analyzed. The influence of heavy metal ions from the raw materials and the COSGB mixtures on the groundwater environment was studied by leaching tests. The research demonstrates that the optimal mix ratio is GGBS mixing amount 4.85%, mass ratio of SS to OSR 0.82, and solid mass concentration 67.69%. At shorter curing age, the hydration products are mainly calcium alumino silicate hydrate (C-A-S-H) and calcium silicate hydrate (C-S-H) gels. With the increase of curing age, ettringite (AFt) and C-S-H gels become the main source of the UCS. Meanwhile, the porosity of the filler decreases continuously. The leaching concentration of heavy metal ions from the COSGB mixtures is all lower than the leaching concentration of raw materials and meet the requirements of the Chinese groundwater quality standard (GB/T 14848-2017). Therefore, this new COSGB cannot pollute the groundwater environment and meets backfill requirements. The proposed technology is a reliable and environmentally friendly alternative for recycling OSR and SS while simultaneously supporting cemented paste backfill (CPB).