Animal models of mpox virus infection and disease.

Mpox (monkeypox) virus (MPXV), which causes a mild smallpox-like disease, has been endemic in Africa for several decades, with sporadic cases occurring in other parts of the world. However, the most recent outbreak of mpox mainly among men that have sex with men has affected several continents, posing serious global public health concerns. The infections exhibit a wide spectrum of clinical presentation, ranging from asymptomatic infection to mild, severe disease, especially in immunocompromised individuals, young children, and pregnant women. Some therapeutics and vaccines developed for smallpox have partial protective and therapeutic effects against MPXV historic isolates in animal models. However, the continued evolution of MPXV has produced multiple lineages, leading to significant gaps in the knowledge of their pathogenesis that constrain the development of targeted antiviral therapies and vaccines. MPXV infections in various animal models have provided a central platform for identification and comparison of diseased pathogenesis between the contemporary and historic isolates. In this review, we discuss the susceptibility of various animals to MPXV, and describe the key pathologic features of rodent, rabbit and nonhuman primate models. We also provide application examples of animal models in elucidating viral pathogenesis and evaluating effectiveness of vaccine and antiviral drugs. These animal models are essential to understand the biology of MPXV contemporary isolates and to rapidly test potential countermeasures. Finally, we list some remaining scientific questions of MPXV that can be resolved by animal models.

14-3-3ζ inhibits maladaptive repair in renal tubules by regulating YAP and reduces renal interstitial fibrosis.

Acute kidney injury (AKI) refers to a group of common clinical syndromes characterized by acute renal dysfunction, which may lead to chronic kidney disease (CKD), and this process is called the AKI-CKD transition. The transcriptional coactivator YAP can promote the AKI-CKD transition by regulating the expression of profibrotic factors, and 14-3-3 protein zeta (14-3-3ζ), an important regulatory protein of YAP, may prevent the AKI-CKD transition. We established an AKI-CKD model in mice by unilateral renal ischemia-reperfusion injury and overexpressed 14-3-3ζ in mice using a fluid dynamics-based gene transfection technique. We also overexpressed and knocked down 14-3-3ζ in vitro. In AKI-CKD model mice, 14-3-3ζ expression was significantly increased at the AKI stage. During the development of chronic disease, the expression of 14-3-3ζ tended to decrease, whereas active YAP was consistently overexpressed. In vitro, we found that 14-3-3ζ can combine with YAP, promote the phosphorylation of YAP, inhibit YAP nuclear translocation, and reduce the expression of fibrosis-related proteins. In an in vivo intervention experiment, we found that the overexpression of 14-3-3ζ slowed the process of renal fibrosis in a mouse model of AKI-CKD. These findings suggest that 14-3-3ζ can affect the expression of fibrosis-related proteins by regulating YAP, inhibit the maladaptive repair of renal tubular epithelial cells, and prevent the AKI-CKD transition.

Synergistic photothermal-photodynamic-chemotherapy toward breast cancer based on a liposome-coated core-shell AuNS@NMOFs nanocomposite encapsulated with gambogic acid.

A multifunctional nanoplatform with core-shell structure was constructed in one-pot for the synergistic photothermal, photodynamic, and chemotherapy against breast cancer. In the presence of gambogic acid (GA) as the heat-shock protein 90 (HSP90) inhibitor and the gold nanostars (AuNS) as the photothermal reagent, the assembly of Zr4+ with tetrakis (4-carboxyphenyl) porphyrin (TCPP) gave rise to the nanocomposite AuNS@ZrTCPP-GA (AZG), which in turn, further coated with PEGylated liposome (LP) to enhance the stability and biocompatibility, and consequently the antitumor effect of the particle. Upon cellular uptake, the nanoscale metal - organic framework (NMOF) of ZrTCPP in the resulted AuNS@ZrTCPP-GA@LP (AZGL) could be slowly degraded in the weak acidic tumor microenvironment to release AuNS, Zr4+, TCPP, and GA to exert the synergistic treatment of tumors via the combination of AuNS-mediated mild photothermal therapy (PTT) and TCPP-mediated photodynamic therapy (PDT). The introduction of GA serves to reduce the thermal resistance of the cell to re-sensitize PTT and the constructed nanoplatform demonstrated remarkable anti-tumor activity in vitro and in vivo. Our work highlights a facile strategy to prepare a pH-dissociable nanoplatform for the effective synergistic treatment of breast cancer.

A new nairo-like virus associated with human febrile illness in China.

ABSTRACTSeveral nairo-like viruses have been discovered in ticks in recent years, but their relevance to public health remains unknown. Here, we found a patient who had a history of tick bite and suffered from a febrile illness was infected with a previously discovered RNA virus, Beiji nairovirus (BJNV), in the nairo-like virus group of the order Bunyavirales. We isolated the virus by cell culture assay. BJNV could induce cytopathic effects in the baby hamster kidney and human hepatocellular carcinoma cells. Negative-stain electron microscopy revealed enveloped and spherical viral particles, morphologically similar to those of nairoviruses. We identified 67 patients as BJNV infection in 2017-2018. The median age of patients was 48 years (interquartile range 41-53 years); the median incubation period was 7 days (interquartile range 3-12 days). Most patients were men (70%), and a few (10%) had underlying diseases. Common symptoms of infected patients included fever (100%), headache (99%), depression (63%), coma (63%), and fatigue (54%), myalgia or arthralgia (45%); two (3%) patients became critically ill and one died. BJNV could cause growth retardation, viremia and histopathological changes in infected suckling mice. BJNV was also detected in sheep, cattle, and multiple tick species. These findings demonstrated that the newly discovered nairo-like virus may be associated with a febrile illness, with the potential vectors of ticks and reservoirs of sheep and cattle, highlighting its public health significance and necessity of further investigation in the tick-endemic areas worldwide.

Identification of a new orthonairovirus associated with human febrile illness in China.

The genus Orthonairovirus, which is part of the family Nairoviridae, includes the important tick-transmitted pathogens Crimean-Congo hemorrhagic fever virus and Nairobi sheep disease virus, as well as many other poorly characterized viruses found in ticks, birds and mammals1,2. In this study, we identified a new orthonairovirus, Songling virus (SGLV), from patients who reported being bitten by ticks in Heilongjiang Province in northeastern China. SGLV shared similar genomic and morphological features with orthonairoviruses and phylogenetically formed a unique clade in Tamdy orthonairovirus of the Nairoviridae family. The isolated SGLV induced cytopathic effects in human hepatoma cells in vitro. SGLV infection was confirmed in 42 hospitalized patients analyzed between 2017 and 2018, with the main clinical manifestations being headache, fever, depression, fatigue and dizziness. More than two-thirds (69%) of patients generated virus-specific antibody responses in the acute phase. Taken together, these results suggest that this newly discovered orthonairovirus is associated with human febrile illness in China.

Magnetic resonance imaging of the zone of calcified cartilage in the knee joint using 3-dimensional ultrashort echo time cones sequences.

BACKGROUND: The zone of calcified cartilage (ZCC) plays an important role in the pathogenesis of osteoarthritis (OA) but has never been imaged in vivo with magnetic resonance (MR) imaging techniques. We investigated the feasibility of direct imaging of the ZCC in both cadaveric whole knee specimens and in vivo healthy knees using a 3-dimensional ultrashort echo time cones (3D UTE-Cones) sequence on a clinical 3T scanner. METHODS: In all, 12 cadaveric knee joints and 10 in vivo healthy were collected. At a 3T MR scanner with an 8-channel knee coil, a fat-saturated 3D dual-echo UTE-Cones sequence was used to image the ZCC, following with a short rectangular pulse excitation and 3D spiral sampling with conical view ordering. The regions of interests (ROIs) were delineated by a blinded observer. Single-component T2* and T2 values were calculated from fat-saturated 3D dual-echo UTE-Cones and a Carr-Purcell-Meiboom-Gill (T2 CPMG) data using a semi-automated MATLAB code. RESULTS: The single-exponential fitting curve of ZCC was accurately obtained with R2 of 0.989. For keen joint samples, the ZCC has a short T2* ranging from 0.62 to 2.55 ms, with the mean ±â€Šstandard deviation (SD) of 1.49 ±â€Š0.66 ms, and with 95% confidence intervals (CI) of 1.20-1.78 ms. For volunteers, the short T2* ranges from 0.93 to 3.52 ms, with the mean ±â€ŠSD of 2.09 ±â€Š0.56 ms, and the 95% CI is 1.43 to 2.74 ms in ZCC. CONCLUSIONS: The high-resolution 3D UTE-Cones sequence might be used to directly image ZCC in the human knee joint on a clinical 3T scanner with a scan time of more than 10 min. Using this non-invasive technique, the T2* relaxation time of the ZCC can be further detected.

Relationship of distraction rate with inferior alveolar nerve degeneration-regeneration shift.

Distraction osteogenesis is an important technique for the treatment of maxillofacial abnormities and defects. However, distraction osteogenesis may cause the injury of the inferior alveolar nerve. The relationship between distraction rate and nerve degeneration-regeneration shift remains poorly understood. In this study, 24 rabbits were randomly divided into four groups. To establish the rabbit mandibular distraction osteogenesis model, the mandibles of rabbits in distraction osteogenesis groups were subjected to continuous osteogenesis distraction at a rate of 1.0, 1.5 and 2.0 mm/d, respectively, by controlling rounds of screwing each day in the distractors. In the sham group, mandible osteotomy was performed without distraction. Pin-prick test with a 10 g blunt pin on the labium, histological and histomorphometric analyses with methylene blue staining, Bodian's silver staining, transmission electron microscopy and myelinated fiber density of inferior alveolar nerve cross-sections were performed to assess inferior alveolar nerve conditions. At 28 days after model establishment, in the pin-prick test, the inferior alveolar nerve showed no response in the labium to a pin pricks in the 2 mm/d group, indicating a severe dysfunction. Histological and histomorphometric analyses indicated that the inferior alveolar nerve suffered more degeneration and injuries at a high distraction rate (2 mm/d). Importantly, the nerve regeneration, indicated by newborn Schwann cells and axons, was more abundant in 1.0 and 1.5 mm/d groups than in 2.0 mm/d group. We concluded that the distraction rate was strongly associated with the inferior alveolar nerve function, and the distraction rates of 1.0 and 1.5 mm/d had regenerative effects on the inferior alveolar nerve. This study provides an experimental basis for the relationship between distraction rate and nerve degeneration-regeneration shift during distraction osteogenesis, and may facilitate reducing nerve complications during distraction osteogenesis.

Detection of Repair of the Zone of Calcified Cartilage with Osteoarthritis through Mesenchymal Stem Cells by Ultrashort Echo Time Magnetic Resonance Imaging.

OBJECTIVE: Currently, magnetic resonance imaging (MRI) is the most commonly used imaging modality for observing the growth and development of mesenchymal stem cells (MSCs) after in vivo transplantation to treat osteoarthritis (OA). However, it is a challenge to accurately monitor the treatment effects of MSCs in the zone of calcified cartilage (ZCC) with OA. This is especially true in the physiological and biochemical views that are not accurately detected by MRI contrast agents. In contrast, ultrashort time echo (UTE) MRI has been shown to be sensitive to the presence of the ZCC, creating the potential for more effectively observing the repair of the ZCC in OA by MSCs. A special focus is given to the outlook of the use of UTE MRI to detect repair of the ZCC with OA through MSCs. The limitations of the current techniques for clinical applications and future directions are also discussed. DATA SOURCES: Using the combined keywords: "osteoarthritis", "mesenchymal stem cells", "calcified cartilage", and "magnetic resonance imaging", the PubMed/MEDLINE literature search was conducted up to June 1, 2017. STUDY SELECTION: A total of 132 published articles were initially identified citations. Of the 132 articles, 48 articles were selected after further detailed review. This study referred to all the important English literature in full. RESULTS: In contrast, UTE MRI has been shown to be sensitive to the presence of the ZCC, creating the potential for more effectively observing the repair of the ZCC in OA by MSCs. CONCLUSIONS: The current studies showed that the ZCC could be described in terms of its histomorphology and biochemistry by UTE MRI. We prospected that UTE MRI has been shown the potential for more effectively observing the repair of the ZCC in OA by MSCs in vivo.

Saikosaponin D relieves unpredictable chronic mild stress induced depressive-like behavior in rats: involvement of HPA axis and hippocampal neurogenesis.

RATIONALE: Saikosaponin D (SSD), a major bioactive component isolated from Radix Bupleuri, has been reported to exert neuroprotective properties. OBJECTIVES: The present study was designed to investigate the anti-depressant-like effects and the potential mechanisms of SSD. METHODS: Behavioural tests including sucrose preference test (SPT), open field test (OFT) and forced swim test (FST) were performed to study the antidepressant-like effects of SSD. In addition, we examined corticosterone and glucocorticoid receptor (GR) levels to evaluate hypothalamic-pituitary-adrenal (HPA) axis function. Furthermore, hippocampal neurogenesis was assessed by testing doublecortin (DCX) levels, and neurotrophic molecule levels were also investigated in the hippocampus of rats. RESULTS: We found that unpredictable chronic mild stress (UCMS) rats displayed lost body weight, decreased sucrose consumption in SPT, reduced locomotive activity in OFT, and increased immobility time in FST. Chronic treatment with SSD (0.75, 1.50 mg/kg) remarkably ameliorated the behavioral deficiency induced by UCMS procedure. SSD administration downregulated elevated serum corticosterone levels, as well as alleviated the suppression of GR expression and nuclear translocation caused by UCMS, suggesting that SSD is able to remit the dysfunction of HPA axis. In addition, Western blot and immunohistochemistry analysis showed that SSD treatment significantly increased the generation of neurons in the hippocampus of UCMS rats indicated by elevated DCX levels. Moreover, hippocampal neurotrophic molecule levels of UCMS rats such as phosphorylated cAMP response element binding protein (p-CREB) and brain-derived neurotrophic factor (BDNF) were raised after SSD treatment. CONCLUSIONS: Together, Our results suggest that SSD opposed UCMS-induced depressive behaviors in rats, which was mediated, partially, by the enhancement of HPA axis function and consolidation of hippocampal neurogenesis.

Role of Diffusion-weighted and Contrast-enhanced Magnetic Resonance Imaging in Differentiating Activity of Ankylosing Spondylitis.

BACKGROUND: Previous studies showed that combining apparent diffusion coefficient (ADC) value with the Spondyloarthritis Research Consortium of Canada (SPARCC) index value might provide a reliable evaluation of the activity of ankylosing spondylitis (AS), and that contrast-enhanced (CE) magnetic resonance imaging (MRI) is unnecessary. However, the results were based on confirming only a small random sample. This study aimed to assess the role of CE-MRI in differentiating the disease activity of AS by comparing ADC value with a large sample. METHODS: A total of 115 patients with AS were enrolled in accordance with Bath AS Disease Activity Index and laboratory indices, and 115 patients were divided into two groups, including active group (n = 69) and inactive group (n = 46). SPARCC, ΔSI, and ADC values were obtained from the short tau inversion recovery (STIR), diffusion-weighted imaging (DWI), and CE-MRI, respectively. One-way analysis of variance and receiver operating characteristic analysis were performed for all parameters. RESULTS: The optimal cutoff values (with sensitivity, specificity, respective area under the curve, positive likelihood ratio, and negative likelihood ratio) for the differentiation between active and inactive groups are as follows: SPARCC = 6 (72.06%, 82.61%, 0.836, 4.14, 0.34); ΔSI (%) = 153 (80.6%, 84.78%, 0.819, 5.3, 0.23); ADC value = 1.15 × 10-3 mm2/s (72.73%, 81.82%, 0.786, 4, 0.33). No statistical differences were found among the predictive values of SPARCC, ΔSI, and ADC. Multivariate analysis showed no significant difference between the combination of SPARCC and ADC values with and without ΔSI. CONCLUSIONS: Using large sample, we concluded that the combination of STIR and DWI would play significant roles in assessing the disease activity, and CE-MRI sequence is not routinely used in imaging of AS to avoid renal fibrosis and aggravation of kidney disease.

Detection of Active Sacroiliitis with Ankylosing Spondylitis through Intravoxel Incoherent Motion Diffusion-Weighted MR Imaging.

OBJECTIVE: To confirm feasibility and assess intravoxel incoherent motion (IVIM) to differentiate active sacroiliitis and ankylosing spondylitis.. METHODS: Forty-one patients were divided into two groups, an active group (n = 20) and a chronic group (n = 21), according to the Bath Ankylosing Spondylitis (AS) Disease Activity Index (BASDAI) and laboratory parameters. In addition, 21 healthy volunteers were chosen as the control group. Tissue diffusivity (Dslow), perfusion fraction (f), and pseudo-diffusion coefficient (Dfast) values were obtained for all three groups. One-way analysis of variance and receiver operating characteristic analysis were performed for all parameters. RESULTS: There was good interobserver agreement on the measurements between the two observers. The optimal cut-off values (with respective AUC, sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio) between active and chronic groups were Dslow = 0.53 × 10(-3) mm(2)/s (0.976, 90%, 95.2%, 18.9, 0.10) and f = 0.09 (0.545, 20%, 95.5%, 4.2, 0.84), and between chronic and control groups were Dslow = 0.22 × 10(-3) mm(2)/s (0.517, 9.52%, 100%, no number, 0.9) and f = 0.09 (0.935, 95.24%, 80.95%, 5, 0.059). CONCLUSION: Dslow and f of IVIM diffusion-weighted (DW)-MRI in AS show a significant difference in the values of diffusion of water molecules and fractional perfusion-related volume among the three groups. KEY POINTS: • D slow can be used to differentiate the activity of AS. • With perfusion fraction, the sensitivity of differentiating the AS activity is improved. • IVIM DWI plays an important role in detecting the activity in patients with AS.

Local injection of substance P increases bony formation during mandibular distraction osteogenesis in rats.

Substance P is a neuropeptide that is distributed in those sensory nerve fibres that innervate the medullary tissues of bone. It is a potent accelerator of proliferation and differentiation of osteoblasts in vitro. However, its capacity for promoting repair of mandibular defects is not known. We have investigated the osteogenic effects of local injections of substance P during mandibular distraction osteogenesis in rats. Twenty Sprague-Dawley rats were randomly assigned to 2 groups (n = 10 in each): substance P 10(-7) mmol/l in normal saline 0.2ml was injected into the experimental group, and saline alone into the controls. The mandibular distraction rate was 0.2mm every 12hours for 10 days. Daily injections of substance P or saline were given during the distraction period. Regeneration of bone was assessed quantitatively on days 15 and 29 using microcomputed tomography (microCT), and histological analysis. The rate of bony union in the group treated with substance P was significantly higher than that in the saline alone group on day 29 (p=0.001) The microCT images and quantitation showed more callus and more mature cortical bone when substance P was given than with control. Histological examination showed that cartilaginous tissues had formed in the middle of the distraction gaps in both groups. Bony bridges were seen only in the substance P group at the final time point (day 29). Injection of substance P into the gap of a rat mandible during mandibular distraction improved formation of good-quality bone and accelerated bony union.

Stability of the recombinant anti­erbB2 scFv­Fc­interleukin­2 fusion protein and its inhibition of HER2­overexpressing tumor cells.

The anti­erbB2 scFv­Fc­IL­2 fusion protein (HFI) is the basis for development of a novel targeted anticancer drug, in particular for the treatment of HER2­positive cancer patients. HFI was fused with the anti­erbB2 antibody and human IL­2 by genetic engineering technology and by antibody targeting characteristics of HFI. IL­2 was recruited to target cells to block HER2 signaling, inhibit or kill tumor cells, improve the immune capacity, reduce the dose of antibody and IL­2 synergy. In order to analyse HFI drug ability, HFI plasmid stability was verified by HFI expression of the trend of volume changes. Additionally, HFI could easily precipitate and had progressive characteristics and thus, the buffer system of the additive phosphate­citric acid buffer, arginine, Triton X­100 or Tween­80, the establishment of a microfiltration, ion exchange, affinity chromatography and gel filtration chromatography­based purification process were explored. HFI samples were obtained according to the requirements of purity, activity and homogeneity. In vivo, HFI significantly delayed HER2 overexpression of non­small cell lung cancer (Calu­3) in human non­small cell lung cancer xenografts in nude mice, and the inhibition rate was more than 60% (P<0.05) in the group treated with 1 mg/kg the HFI dose; HFI significantly inhibited HER2 expression of breast cancer (FVB/neu) transgenic mouse tumor growth in 1 mg/kg of the HFI dose group, and in the following treatment the 400 mm3 tumors disappeared completely. Combined with other HFI test data analysis, HFI not only has good prospects, but also laid the foundation for the development of antibody­cytokine fusion protein­like drugs.

Effects of breviscapine on pulmonary inflammatory response and lung injury in children undergoing open heart surgery.

This study examined the effects of breviscapine (1) on pulmonary inflammatory response and lung function in pediatric patients undergoing open heart surgery. Forty-five children (ASA II or III, aged 2-72 months) were randomly assigned to control group (saline, Group C), low dose 1 group (0.5 mg/kg, Group Bre0.5), and high dose 1 group (1.0 mg/kg, Group Bre1.0), 15 cases each group. Plasma concentrations of procalcitonin (PCT) and neutrophil elastase (NE) were measured and compared at different time points. Plasma concentrations of PCT and NE were increased after cardiopulmonary bypass (CPB) induction, and the concentrations were lower in 1-treated groups. The present results indicated that continuous infusion of 1 before the CPB suppressed the production of PCT and NE attenuated systemic inflammatory response, which could result in lung protective effect in children undergoing open heart surgery.

Effects of nerve growth factor delivery via a gel to inferior alveolar nerve in mandibular distraction osteogenesis.

Inferior alveolar nerve (IAN) injury is a concern in mandible distraction osteogenesis (DO). We have previously demonstrated that repeated local injections of human nerve growth factor beta (NGF-beta) have significantly enhanced the histologic recovery of the IAN in a rabbit model of DO. This study was to further test the effect of a single injection of human NGF-beta delivered via a collagen/nanohydroxyapatite/kappa-carrageenan gel to the recovery of the IAN in DO. Rabbits underwent mandibular DO at a rate of 0.75 mm/12 h for 6 days. At the end of the distraction period, injections were performed near the IAN percutaneously as follows: group 1, human NGF-beta in the gel; group 2, human NGF-beta in saline; group 3, the gel alone; and group 4, saline alone. At 14 days after the end of distraction, IAN histologic findings and histomorphometric parameters were evaluated. Histologically, there were less myelin debris and more abundant regenerating nerve fibers in group 1 than the other groups. Both the myelinated fiber density and the myelinated axon area in group 1 were significantly higher than groups 3 and 4 (P < 0.01); the myelinated axon area in the group 1 was significantly higher than group 2 (P < 0.01). In conclusion, the delivery of human NGF-beta in the gel leads to a better acceleration of the IAN injury recovery over the saline delivery. It provides a possible way to enhance the recovery of nerve injuries in craniofacial DO clinically.

[Machinable property of a novel dental mica glass-ceramic].

OBJECTIVE: To investigate the machinability of a novel dental mica glass-ceramic and analyze the effect of heat-treatment on its ductile machinable behavior. METHODS: The drilling and turning experiment were used to measure the machinabilities of the control group (feldspar ceramic: Vita Mark II) and 7 experiment groups treated with different crystallization techniques. The microstructures were analyzed using scanning electron microscopy (SEM) and X-ray diffraction (XRD). RESULTS: The average drilling depths in 30 s of the experimental groups ranged from (0.5 +/- 0.1) mm to (7.1 +/- 0.8) mm. There were significant differences between the control [(0.8 +/- 0.1) mm] and the experimental groups (P < 0.05) except the group crystallized at 740 degrees C for 60 min. When crystallized at 650 degrees C (60 min), continuous band chips could formed in machining at a high velocity and cut depth. The crystal portion of this group is only about 40%. CONCLUSIONS: This material has a satisfactory machinability. The mechanism could be attributed to a combination of the interlocked structure of mica crystals and the low viscosity of glassy phase.

Lead zinc borate, PbZn2(BO3)2.

PbZn2(BO3)2 crystallizes in the space group Pccn, with the Pb cation at a site with imposed twofold symmetry. The compound represents a new structure type in which ZnBO3 layers are bridged by Pb2+ cations, giving rise to a three-dimensional framework. Channels parallel to the [010] direction accommodate the stereochemically active lone pairs of the Pb2+ cations.