Hematuria after nocturnal exercise of a man: A case report.

BACKGROUND: A man experienced multiple episodes of macroscopic hematuria following nocturnal exercise. Urinary stones and tumors were considered the two most likely causes. The patient had two hobbies: Consuming health care products in large quantities and engaging in late-night running. CASE SUMMARY: Health care products contain a large amount of calcium phosphate, and we hypothesize that this could induce the formation of small phosphate stones. After exercise, the urinary system is abraded, resulting in bleeding. The patient was advised to stop using the health care products. Consequently, the aforementioned symptoms disappeared immediately. However, the patient resumed the above two habits one year later; correspondingly, the macroscopic hematuria reappeared. CONCLUSION: This finding further confirmed the above inference and allowed for a new avenue to determine the cause of the patient's hematuria.

High hemodynamic stresses induce aneurysms at internal carotid artery bends.

To investigate the role of hemodynamic stresses in initiating cerebral aneurysms at bends of internal carotid artery (ICA). Sixty-one patients with 68 aneurysms at ICA bends were retrospectively enrolled as the experiment group. Among the 61 patients, 30 normal ICAs without aneurysms were chosen as the control. All patients had 3-dimensional angiography and CFD analysis. The bending angle was significantly (P < .0001) smaller in the experiment than control group (131.2º ± 14.9º vs 150.3º ± 9.5º). The dynamic pressure, shear stress, vorticity magnitude and strain rate were the least at direct flow impinging center where the total pressure was very high. The dynamic stress, shear stress, strain rate and gradients of total pressure except for gradient 1 were significantly (P < .05) greater at the aneurysm site than at all the other sites. The total pressure at the aneurysm site was greater (P < .05) than at 1 lateral location and at the distal area but smaller (P < .05) than at the proximal area. The dynamic pressure, shear stress, strain rate and gradient of total pressure at the aneurysm site were significantly (P < .001) greater than on the aneurysm dome. The hemodynamic stresses were all significantly (P < .01) greater at the aneurysm site in the experiment group than at the site corresponding to the aneurysm in the control group. Aneurysms at the ICA bends are caused by direct flow impingement and increased hemodynamic stresses, and smaller arterial bending angles result in abnormally enhanced hemodynamic stresses to initiate an aneurysm near the flow impingement area.

Transcriptome analysis of sweet potato responses to potassium deficiency.

BACKGROUND: As one of three essential nutrients, potassium is regarded as a main limiting factor for growth and development in plant. Sweet potato (Ipomoea batatas L.) is one of seven major food crops grown worldwide, and is both a nutrient-rich food and a bioenergy crop. It is a typical 'K-favoring' crop, and the level of potassium ion (K+) supplementation directly influences its production. However, little is known about the transcriptional changes in sweet potato genes under low-K+ conditions. Here, we analyzed the transcriptomic profiles of sweet potato roots in response to K+ deficiency to determine the effect of low-K+ stress on this economically important crop. RESULTS: The roots of sweet potato seedlings with or without K+ treatment were harvested and used for transcriptome analyses. The results showed 559 differently expressed genes (DEGs) in low and high K+ groups. Among the DEGs, 336 were upregulated and 223 were downregulated. These DEGs were involved in transcriptional regulation, calcium binding, redox-signaling, biosynthesis, transport, and metabolic process. Further analysis revealed previously unknow genes involved in low-K+ stress, which could be investigated further to improve low K+ tolerance in plants. Confirmation of RNA-sequencing results using qRT-PCR displayed a high level of consistency between the two experiments. Analysis showed that many auxin-, ethylene- and jasmonic acid-related genes respond to K+ deficiency, suggesting that these hormones have important roles in K+ nutrient signaling in sweet potato. CONCLUSIONS: According to the transcriptome data of sweet potato, various DEGs showed transcriptional changes in response to low-K+ stress. However, the expression level of some kinases, transporters, transcription factors (TFs), hormone-related genes, and plant defense-related genes changed significantly, suggesting that they have important roles during K+ deficiency. Thus, this study identifies potential genes for genetic improvement of responses to low-K+ stress and provides valuable insight into the molecular mechanisms regulating low K+ tolerance in sweet potato. Further research is required to clarify the function of these DEGs under low-K+ stress.

Endovascular Retreatment of Cerebral Aneurysms Previously Treated with Endovascular Embolization.

OBJECTIVE: Intracranial aneurysms treated with endovascular coil embolization may recur. We investigated the factors affecting aneurysmal recurrence after embolization and effects of endovascular retreatment within 1 year. METHODS: In 3 years, 1,335 patients with 1,385 intracranial aneurysms were treated with coil embolization. Factors affecting aneurysm recurrence and the effects of endovascular retreatment were analyzed. RESULTS: Angiography immediately following embolization showed total occlusion in 1,030 aneurysms (74.4%), neck remnant in 207 (14.9%), and partial occlusion in 148 (10.7%), with a total peri-procedure complication rate of 4.2%. Overall, 145 patients with 151 aneurysms recurred within 1 year and the other 1,234 aneurysms remained occluded (89.1%). A significant (p < 0.05) difference existed in aneurysm size, rupture status, use of stent and immediate occlusion outcome between the two groups, with significantly (p < 0.05) lower recurrence rates in aneurysms with smaller sizes, no rupture and stent-assistance coiling. Neck remnant, partial occlusion, coiling without stent assistance, large and giant aneurysms were significant (p < 0.05) risk factors for aneurysm recurrence during the first year. The rate of recurrence was 4.7% (11/232) in aneurysms with total occlusion and 35.9% (23/64) in aneurysms with neck remnant and partial occlusion. Of the 34 recurrent aneurysms, 6 were re-embolized with detachable coils alone, 12 with stent-assisted coiling, 8 with balloon-assisted embolization, and the remaining 8 aneurysms with covered stents, resulting in total occlusion in 28 aneurysms and neck remnant in 6. CONCLUSION: Recurrence of previously-coiled cerebral aneurysms is significantly affected by aneurysm size, use of stent and degree of immediate occlusion. Endovascular retreatment with balloon-or stent-assisted techniques or with covered stents can be safe and effective for recurrent cerebral aneurysms.

Can we predict who will develop postoperative hyperkalaemia after parathyroidectomy in dialysis patients with secondary hyperparathyroidism?

BACKGROUND: Hyperkalaemia occurs frequently in many maintenance haemodialysis (MHD) patients after parathyroidectomy (PTX) with secondary hyperparathyroidism (SHPT). However, the clinical risk factors that predict postoperative hyperkalaemia are uncertain. METHODS: This retrospective cohort study included 90 maintenance haemodialysis patients aged ≥18 years who underwent PTX between April 2011 and April 2016 at Aerospace Center Hospital (Peking University Aerospace School of Clinical Medicine). Pre- and post-PTX surgery venous samples were measured in quadruplicate. We examined univariate associations with demographics, dialysis characteristics, laboratory values and medications. Hyperkalaemia was defined as serum potassium >5.3 mmol/L. RESULTS: Out of nighty patients, twenty-two (24.4%) developed postoperative hyperkalaemia, of whom sixteen (18.1%) developed hyperkalaemia on postoperative day 3. The univariate analysis showed that weight, dialysis duration, preoperative serum potassium, alkaline phosphate, triglyceride, and postoperative alkaline phosphate were independently associated with hyperkalaemia after parathyroidectomy. The univariate logistic regression model showed that preoperative serum potassium was the only independent factor that could predict hyperkalaemia after parathyroidectomy (odds ratio, 1.59; 95% confidence interval, 1.24-2.05). The optimal cut-off for pre-operative K was 3.9 mmol/L according to the receiver operating characteristic (ROC) curve. A higher incidence of postoperative hyperkalaemia was found in male and younger patients, but the difference was not statistically significant (p>0.05). CONCLUSIONS: Pre-operative serum potassium less than 3.9 mmol/L was associated with less hyperkalaemia post-operatively in end-stage renal disease (ESRD) patients undergoing PTX.

Construction of Aneurysmal Models on a Curved Vascular Segment of a Carotid Siphon Model for Testing Endovascular Devices.

OBJECTIVE: To investigate construction of an aneurysm on a curved vascular segment of a carotid siphon model for testing endovascular devices. METHODS: Preshaped carotid siphon models of polytetrafluoroethylene were constructed from a human cadaver for confining canine common carotid artery (CCA). The canine right external jugular vein was isolated and harvested to make a venous pouch by suturing 1 end. The right CCA was isolated, and the venous pouch was sutured onto the right CCA to make an aneurysm. The right CCA segment containing the aneurysm was excised and guided through the preshaped polytetrafluoroethylene carotid siphon model using a guidewire with the aneurysm adjusted to protrude out of the round window of the siphon model. The siphon model together with the aneurysm was sutured end-to-end onto the left CCA to form a carotid siphon model in vivo. RESULTS: Five canine models were successfully constructed; the average construction time was 120 minutes. All aneurysms and siphon models remained patent 7 days and 2 weeks later. Five covered stents for intracranial use were tested for flexibility and apposition to the vascular wall in the curved segment of the carotid siphon model in vivo. All the covered stents passed the tortuous siphon model without much difficulty and were deployed successfully to cover the aneurysm orifice without endoleak. CONCLUSIONS: The carotid siphon model in vivo can simulate well the geometry of the human carotid siphon segment and can be used to test endovascular devices for interventions.

Hemodynamic Factors Affecting Carotid Sinus Atherosclerotic Stenosis.

PURPOSE: Carotid atherosclerotic plaque occurs predominantly at the outer wall of the carotid sinus, and computational fluid dynamics (CFD) plays an important role in explaining plaque formation. The present study investigated the hemodynamic factors affecting carotid atherosclerotic stenosis. METHODS: Sixteen patients with normal carotid arteries and 16 with symptomatic stenotic carotid sinus underwent 3-dimensional angiographic imaging evaluations and were studied with CFD to simulate the complete 3-dimensional blood flow and hemodynamic parameter distribution in the carotid bifurcations. The hemodynamic parameters, including wall shear stress (WSS), dynamic and total pressure, total pressure gradient, strain rate, velocity, and velocity angle, were investigated. RESULTS: The atherosclerosis-prone outer lateral walls of the carotid sinus and the external carotid artery at its start had significantly (P < 0.05) low dynamic pressure, WSS, strain rate, and total pressure gradient but high static pressure. The blood flow near these walls with flow separation had significantly (P < 0.05) decreased velocity and dynamic pressures but a high velocity angle. The carotid divider had significantly (P < 0.05) elevated dynamic and total pressure, WSS, strain rate, and total pressure gradient but reduced static pressure. Additional stenosis occurred at the downstream area of stenosis with significantly (P < 0.05) decreased dynamic pressure, WSS, strain rate, and total pressure gradient similar to the wall at the sinus and the start of the external carotid artery. CONCLUSION: Significantly decreased vascular WSS, dynamic pressure, strain rate, and total pressure gradient are key to atherosclerotic plaque formation at the carotid sinus.

RETRACTED: Endovascular Stent Deployment in the Management of Lesions Related to Internal Carotid Artery Redundancy.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of of the Editor-in-Chief because this article has included both figures and data from patients that were cared for by Dr. Malek at the Cerebrovascular Hemodynamics laboratory in the Department of Neurosurgery, at Tufts Medical Center. As we understand, Dr. Gao did not have clinical privileges and played no clinical role in the care of Dr. Malek's patients while at Tufts Medical Center.

Small Tenuous Intracranial Arteries Can Well Tolerate the Deployment of 2 Stents in Y Configuration or an Overlapping Manner in Treating Intracranial Aneurysms.

OBJECTIVE: To investigate parent vessel response to deployment of 2 stents for treatment of cerebral aneurysms. METHODS: Fifteen patients (11 women and 4 men; age range, 25-83 years) with 18 wide-necked intracranial aneurysms were treated with 2 stents with or without subsequent coiling. The vascular diameter was measured and compared within the native parent artery, and the single stent and double stent were measured and compared before and immediately after stenting and at angiographic follow-up. RESULTS: Thirty stents were deployed. Before stenting, the mean vessel diameter was 3.4 ± 0.21 mm at point A, 3.06 ± 0.18 mm at point B, 3.16 ± 0.21 mm at point C, 2.67 ± 0.27 mm at point D, and 2.56 ± 0.23 mm at point E. The deployment of 2 stents resulted in statistically significant increases in both the average vascular diameter and cross-sectional area at points C (3.51 ± 0.22 mm, P = 0.0006; and 9.76 ± 1.17 mm2, P = 0.001, respectively) and E (2.88 ± 0.32 mm, P = 0.01; and 7.28 ± 1.46 mm2, P = 0.02, respectively) compared with prestenting. At angiographic follow-ups, compared with before stenting, significant increases were documented at point C (3.42 ± 0.22 mm and 9.42 ± 1.37 mm2, respectively) at first angiographic follow-up but at points A (3.62 ± 0.45 mm and 10.51 ± 2.37 mm2, respectively) and B (3.26 ± 0.24 mm and 8.47±1.26 mm2, respectively) at second angiographic follow-up. No significant vascular stenosis was demonstrated at the double-stent segment compared with the single-stent or native artery segments. CONCLUSIONS: The small tenuous cerebral arteries can well tolerate the deployment of 2 stents for the treatment of intracranial aneurysms.

Comparison of surgical and endovascular approaches in the management of multiple intracranial aneurysms.

OBJECTIVE: To investigate the outcomes and safety of endovascular compared with surgical clipping for multiple intracranial aneurysms. MATERIAL AND METHODS: 98 patients with 260 multiple intracranial aneurysms were treated with endovascular, surgical clipping, combined treatment, and observation. Data were retrospectively studied following treatment and at follow-up. RESULTS: In the endovascular group, 44 aneurysms were treated with coils only and 29 aneurysms were treated with stent deployment. The complete occlusion rate was 65%, and the total complication rate was 12% with no permanent deficit. After angiographic follow-up for 1-90 (mean 62) months, the total recurrence rate was 18.3%. In the clipping group, 65 aneurysms were clipped. The complete occlusion rate was 90.8%, and the complication rate was 10.9% with 1 permanent deficit. After follow-up for 11-71 (mean 49) months, the angiographic recurrence rate was 1.5%. In the combination group, 20 aneurysms were treated endovascularly. The complete occlusion rate was 78.9%, and the complication rate was 15.8% with no permanent deficit. Twenty-eight aneurysms were treated surgically with the complete occlusion rate of 89.3%, the complication rate of 20% and 3 permanent deficits. After follow-up for 1-93 (mean 58) months, the angiographic recurrence rate was 33.3% for embolization and 3.6% for clipping. Seventy-four aneurysms for observation had 2.7% regrowth rate within 1-3 years. CONCLUSION: Endovascular embolization has an accepted complication rate but no neurological deficits compared with surgical clipping and may be a better approach for multiple intracranial aneurysms than surgical clipping.

Genetic diversity and evolutionary dynamics of Ebola virus in Sierra Leone.

A novel Ebola virus (EBOV) first identified in March 2014 has infected more than 25,000 people in West Africa, resulting in more than 10,000 deaths. Preliminary analyses of genome sequences of 81 EBOV collected from March to June 2014 from Guinea and Sierra Leone suggest that the 2014 EBOV originated from an independent transmission event from its natural reservoir followed by sustained human-to-human infections. It has been reported that the EBOV genome variation might have an effect on the efficacy of sequence-based virus detection and candidate therapeutics. However, only limited viral information has been available since July 2014, when the outbreak entered a rapid growth phase. Here we describe 175 full-length EBOV genome sequences from five severely stricken districts in Sierra Leone from 28 September to 11 November 2014. We found that the 2014 EBOV has become more phylogenetically and genetically diverse from July to November 2014, characterized by the emergence of multiple novel lineages. The substitution rate for the 2014 EBOV was estimated to be 1.23 × 10(-3) substitutions per site per year (95% highest posterior density interval, 1.04 × 10(-3) to 1.41 × 10(-3) substitutions per site per year), approximating to that observed between previous EBOV outbreaks. The sharp increase in genetic diversity of the 2014 EBOV warrants extensive EBOV surveillance in Sierra Leone, Guinea and Liberia to better understand the viral evolution and transmission dynamics of the ongoing outbreak. These data will facilitate the international efforts to develop vaccines and therapeutics.

Microcatheter looping facilitates access to both the acutely angled parent artery and cerebral aneurysms for effective embolization.

Aneurysms with an acutely angled parent artery are difficult to access for coiling. This study aimed to investigate the safety and effectiveness of microcatheter looping for embolization of cerebral aneurysms with access difficulty. Ten patients (male:female=5:5) with cerebral aneurysms treated with the microcatheter looping technique were analyzed retrospectively. The parent artery formed an acute angle with the major artery in five aneurysms. The microcatheter was looped into a "α" loop for treatment in the anterior temporal artery aneurysm and a "U" loop in the remaining nine aneurysms. All ten aneurysms were successfully treated with the microcatheter looping technique. The microcatheter tip was successfully navigated into the aneurysm sac and remained stable throughout the embolization process. All aneurysms were occluded with total occlusion in five and near-total occlusion in five, and the parent artery remained patent in all cases. No complications occurred peri-procedurally. The Glasgow Outcome Scale was 5 in all patients before discharge. Follow-up angiography six to 12 months later revealed a good occlusion status of the aneurysms. The microcatheter looping technique is effective when the conventional embolization technique fails to treat cerebral aneurysms with difficult access especially when the parent artery forming an acute angle with the major artery exacerbates difficult access to the aneurysms.

The stent-assisted coil-jailing technique facilitates efficient embolization of tiny cerebral aneurysms.

OBJECTIVE: Tiny cerebral aneurysms are difficult to embolize because the aneurysm's sac is too small for a single small coil, and coils within the aneurysm may escape from the confinement of a stent. This study was performed to introduce the stent-assisted coil-jailing technique and to investigate its effect on the coil embolization of tiny intracranial aneurysms. MATERIALS AND METHODS: Sixteen patients with tiny intracranial aneurysms treated with the stent-assisted coil-jailing technique between January 2011 and December 2013 were retrospectively reviewed and followed-up. RESULTS: All aneurysms were successfully treated with the coil-jailing technique, and at the end of embolization, complete occlusion of the aneurysm was achieved in 9 cases (56.3%), incomplete occlusion in 6 (37.5%), and partial occlusion in 1 (6.3%). Intraprocedural complications included acute thrombosis in one case (6.3%) and re-rupture in another (6.3%). Both complications were managed appropriately with no sequela. Follow-up was performed in all patients for 3-24 months (mean, 7.7 months) after embolization. Complete occlusion was sustained in the 9 aneurysms with initial complete occlusion, progressive thrombosis to complete occlusion occurred in the 6 aneurysms with initial near-complete occlusion, and one aneurysm resulted in progressive thrombosis to complete occlusion after initial partial occlusion. No migration of stents or coils occurred at follow-up as compared with their positions immediately after embolization. At follow-up, all patients had recovered with no sequela. CONCLUSION: The stent-assisted coil-jailing technique can be an efficient approach for tiny intracranial aneurysms, even though no definite conclusion regarding its safety can be drawn from the current data.

Production and characterization of a fusion peptide derived from the rabies virus glycoprotein (RVG29).

Gene therapy targeting the brain holds great promise in curing nervous system degenerative diseases in clinical applications. With this in mind, in a previous study a 29 amino-acid peptide derived from the rabies virus glycoprotein (RVG29) with a nonamer stretch of arginine residues (RVG29-9R) at its carboxy-terminus was exploited as a ligand for brain-targeting gene delivery. Importantly, the report demonstrated that the RVG29-9R vector was able to cross the blood-brain barrier. RVG29-9R is currently synthesized by commercial companies with high associated costs. In this study, in order to reduce the costs of producing RVG29-9R, we have expressed and purified 6mg of a recombinant peptide (RVG29-9R-6His) from 0.4g of cultured Escherichia coli. We assessed the physiochemical properties of RVG29-9R-6His, its cytotoxicity, and the in vitro transfection efficiency in Neuro 2a cells (which express the acetylcholine receptor). Our results reveal that the RVG29-9R-6His peptide recognized Neuro 2a cells in a dose-dependent manner and it was also able to bind plasmid DNA and deliver it into the Neuro 2a cells effectively. Therefore, our study has demonstrated that the recombinant RVG29-9R-6His peptide retains the functions of RVG29-9R and so may provide an economically viable and alternative production method for the manufacture of RVG29-9R.

Aptamers targeting rabies virus-infected cells inhibit street rabies virus in vivo.

Rabies is a viral infection of the CNS that is almost always fatal once symptoms occur. No effective treatment of the disease is available and novel antiviral strategies are urgently required. Street rabies viruses are field isolates known to be highly neurotropic. Aptamers are single-stranded oligonucleotides that bind their targets with high affinity and specificity and thus have potential for use in diagnostic and therapeutic applications. In this study, we demonstrate that the aptamers FO24 and FO21, which target RABV-infected cells, can significantly protect mice from a lethal dose of the street rabies virus FJ strain in vivo. Groups receiving preexposure prophylaxis had higher survival rates than the groups receiving postexposure prophylaxis. When mice were inoculated with aptamers (4 nmol) for 24h by intracranial or intramuscular injection prior to intramuscular inoculation with the FJ strain, approximately 60% of the mice survived. These results indicate that the FO21 and FO24 aptamers may be used to develop preventative antiviral therapy against rabies disease.

An inactivated recombinant rabies CVS-11 virus expressing two copies of the glycoprotein elicits a higher level of neutralizing antibodies and provides better protection in mice.

The rabies virus (RABV) G protein is the primary contributor to the pathogenicity and protective immunity of RABV. In this study, we generated a recombinant rCVS-11-G strain containing two copies of the G protein derived from the pathogenic wild-type (wt) CVS-11 strain and based on its infectious clone. Compared with the wtCVS-11 strain, the rCVS-11-G strain possessed a larger virion and 1.4-fold more G protein, but it exhibited a similar growth property to the rCVS-11 strain, including passaging stability in vitro. qPCR results showed that the two G genes were over-expressed in BHK-21 cells infected with the rCVS-11-G strain. However, the rCVS-11-G strain presented an 80 % lower LD50 than the wtCVS-11 strain when intracranially (i.c.) inoculated in adult mice. Adult mice that were either intracranially (i.c.) or intramuscularly (i.m.) inoculated with rCVS-11-G strain developed more acute neurological symptoms and greater mortality than those inoculated with the wtCVS-11 strain. Furthermore, the rCVS-11-G strain was more easily and rapidly taken up by neuroblastoma cells. These data indicated that the rCVS-11-G strain might have increased neurotropism because of the over-expression of the pathogenic G protein. The inactivated rCVS-11-G strain induced significantly higher levels of virus neutralization antibodies and provided better protection from street rabies virus challenge in mice. Therefore, the rCVS-11-G strain may be a promising inactivated vaccine strain due to its better immunogenicity.

Selection of an aptamer against rabies virus: a new class of molecules with antiviral activity.

Rabies is a fatal central nervous system (CNS) disease caused by the neurotropic rabies virus (RABV). The therapeutic management of RABV infections is still problematic, and novel antiviral strategies are urgently required. We established the RVG-BHK-21 cell line, which expresses RABV glycoprotein on the cell surface, to select aptamers. Through 28 iterative rounds of selection, single-stranded DNA (ssDNA) aptamers were generated by exponential enrichment (SELEX). A virus titer assay and a real-time quantitative reverse transcription PCR (qRT-PCR) assay revealed that four aptamers could inhibit the replication of RABV in cultured baby hamster kidney (BHK)-21 cells. However, the aptamers did not inhibit the replication of other virus, e.g., canine distemper virus (CDV) and canine parvovirus (CPV). In addition, the GE54 aptamer was found to effectively protect mice against lethal RABV challenge. After inoculation with aptamers for 24h or 48h, followed by inoculation with CVS-11, approximately 25-33% of the mice survived. In summary, we selected aptamers that could significantly protect from a lethal dose of RABV in vitro and in vivo.

Canine parvovirus VP2 protein expressed in silkworm pupae self-assembles into virus-like particles with high immunogenicity.

The VP2 structural protein of parvovirus can produce virus-like particles (VLPs) by a self-assembly process in vitro, making VLPs attractive vaccine candidates. In this study, the VP2 protein of canine parvovirus (CPV) was expressed using a baculovirus expression system and assembled into parvovirus-like particles in insect cells and pupae. Electron micrographs of VLPs showed that they were very similar in size and morphology when compared to the wild-type parvovirus. The immunogenicity of the VLPs was investigated in mice and dogs. Mice immunized intramuscularly with purified VLPs, in the absence of an adjuvant, elicited CD4(+) and CD8(+) T cell responses and were able to elicit a neutralizing antibody response against CPV, while the oral administration of raw homogenates containing VLPs to the dogs resulted in a systemic immune response and long-lasting immunity. These results demonstrate that the CPV-VLPs stimulate both cellular and humoral immune responses, and so CPV-VLPs may be a promising candidate vaccine for the prevention of CPV-associated disease.