Inactivation of Tnf-α/Tnfr signaling attenuates progression of intervertebral disc degeneration in mice.

Background: Intervertebral disc degeneration (IVDD) is a major cause of low back pain (LBP), worsened by chronic inflammatory processes associated with aging. Tumor necrosis factor alpha (Tnf-α) and its receptors, Tnf receptor type 1 (Tnfr1) and Tnf receptor type 2 (Tnfr2), are upregulated in IVDD. However, its pathologic mechanisms remain poorly defined. Methods: To investigate the role of Tnfr in IVDD, we generated global Tnfr1/2 double knockout (KO) mice and age-matched control C57BL/6 male mice, and analyzed intervertebral disc (IVD)-related phenotypes of both genotypes under physiological conditions, aging, and lumbar spine instability (LSI) model through histological and immunofluorescence analyses and µCT imaging. Expression levels of key extracellular matrix (ECM) proteins in aged and LSI mice, especially markers of cell proliferation and apoptosis, were evaluated in aged (21-month-old) mice. Results: At 4 months, KO and control mice showed no marked differences of IVDD-related parameters. However, at 21 months of age, the loss of Tnfr expression significantly alleviated IVDD-like phenotypes, including a significant increase in height of the nucleus pulposus (NPs) and reductions of endplates (EPs) porosity and histopathological scores, when compared to controls. Tnfr deficiency promoted anabolic metabolism of the ECM proteins and suppressed ECM catabolism. Tnfr loss largely inhibited hypertrophic differentiation, and, in the meantime, suppressed cell apoptosis and cellular senescence in the annulus fibrosis, NP, and EP tissues without affecting cell proliferation. Similar results were observed in the LSI model, where Tnfr deficiency significantly alleviated IVDD and enhanced ECM anabolic metabolism while suppressing catabolism. Conclusion: The deletion of Tnfr mitigates age-related and LSI-induced IVDD, as evidenced by preserved IVD structure, and improved ECM integrity. These findings suggest a crucial role of Tnf-α/Tnfr signaling in IVDD pathogenesis in mice. Targeting this pathway may be a novel strategy for IVDD prevention and treatment.

Safety and efficacy assessment of an mRNA rabies vaccine in dogs, rodents, and cynomolgus macaques.

Rabies is a lethal disease caused by the rabies virus (RABV), which causes acute neurological infections in mammals, including human beings. We previously reported that an mRNA vaccine (LVRNA001) encoding the rabies virus's glycoprotein induced strong protective immune responses to rabies in mice and dogs. Here, we further evaluate the safety of LVRNA001. First, we performed a confirmative efficacy study in dogs, which showed that LVRNA001 fully protected the animals from the virus, both pre- and post-infection. Moreover, using pre- and post-exposure prophylaxis murine models, we showed that LVRNA001, built from the CTN-1 strain, was able to protect against various representative RABV strains from the China I-VII clades. To evaluate the safety of the vaccine, chronic and reproductive toxicity studies were performed with cynomolgus macaques and rats, respectively. In a repeated-dose chronic toxicity study, vaccinated monkeys displayed no significant alterations in body weight, temperature, or hematological and biochemical markers. Lymphocyte subset measurement and histopathological examination showed that no toxicity was associated with the vaccine. The immunogenicity study in cynomolgus macaques demonstrated that LVRNA001 promoted the generation of neutralizing antibodies and Th1-biased immune response. Evaluation of reproductive toxicity in rats revealed that administration of LVRNA001 had no significant effects on fertility, maternal performance, reproductive processes, and postnatal outcomes. In conclusion, LVRNA001 can provide efficient protection against rabies virus infection in dogs and mice, and toxicity studies showed no significant vaccine-related adverse effects, suggesting that LVRNA001 is a promising and safe vaccine candidate for rabies prophylaxis and therapy.

Advancements in health informatics: finite element insights into medial open-wedge high tibial osteotomy and lateral meniscal tears.

Knee medial compartment osteoarthritis is effectively treated by a medial open-wedge high tibial osteotomy (MOWHTO). The feasibility and safety of MOWHTO for mild lateral meniscal tears are unknown. This study examined the feasibility and safety of knee joint weight-bearing line ratio (WBLr) adjustment during MOWHTO with lateral meniscal injuries. We used a healthy adult male's lower extremities computed tomography scans and knee joint magnetic resonance imaging images to create a normal fine element (FE) model. Based on this model, we generated nine FE models for the MOWHTO operation (WBLr: 40-80%) and 15 models for various lateral meniscal injuries. A compressive load of 650N was applied to all cases to calculate the von Mises stress (VMS), and the intact lateral meniscus' maximal VMS at 77.5% WBLr was accepted as the corrective upper limit stress. Our experimental results show that mild lateral meniscal tears can withstand MOWHTO, while severe tears cannot. Our findings expand the use of MOWHTO and provide a theoretical direction for practical decisions in patients with lateral meniscal injuries.

Highly Sensitive Force Sensor Based on High-Q Asymmetric V-Shaped CaF2 Resonator.

Whispering gallery mode (WGM) resonators have high-quality factors and can be used in high-sensitivity sensors due to the narrow line width that allows for the detection of small external changes. In this paper, a force-sensing system based on a high-Q asymmetric V-shaped CaF2 resonator is proposed. Based on the dispersion coupling mechanism, the deformation of the resonator is achieved by loading force, and the resonant frequency is changed to determine the measurement. By adjusting the structural parameters of the asymmetric V-shaped resonator, the deformation of the resonator under force loading is improved. The experimental results show that the sensitivity of the V-shaped tip is 18.84 V/N, which determines the force-sensing resolution of 8.49 µN. This work provides a solution for force-sensing measurements based on a WGM resonator.

Efficacy of cervical perivascular sympathectomy in improving upper limb motor function in children with cerebral palsy and construction of a predictive model.

BACKGROUND: The effectiveness of cervical perivascular sympathectomy (CPVS) in enhancing upper limb motor function in children with cerebral palsy is unclear, and the factors that influence the effectiveness of the surgery have not been documented. OBJECTIVE: To investigate the effectiveness of CPVS in enhancing upper limb motor function in children with cerebral palsy and develop a predictive chart for potential associated adverse outcomes METHODS: The study included 187 children with cerebral palsy who underwent CPVS at the Cerebral Palsy Center, Second Affiliated Hospital of Xinjiang Medical University, between January 2018 and January 2022. Patients were categorized into two groups based on prognostic outcomes: those with adverse and favorable prognoses. Demographic and laboratory data were collected and analyzed from both groups. To identify independent predictors of poor post-CPVS upper limb motor function outcomes, statistical techniques, including univariate analysis and binary logistic regression, were applied. Subsequently, these predictors were integrated to formulate a comprehensive predictive model. RESULTS: In this cohort of 187 children with cerebral palsy undergoing CPVS, 68 (36.36%) exhibited a favorable prognosis for upper limb motor function and 119 (63.64%) demonstrated an adverse prognosis. Age, motor function, and serum albumin levels were identified as significant prognostic factors via logistic regression analysis. To develop the model, we divided the sample into a training set (70%, n = 131) and a validation set (30%, n = 56). Employing motor function, serum albumin levels, and age as variables, we crafted a predictive model. The model's performance, reflected by the area under the curve was 0.813 (0.732, 0.894) in the training set and 0.770 (0.647, 0.892) in the validation set, demonstrating its robust predictive capability for post-CPVS adverse outcomes. Furthermore, the consistency curve and Hosmer-Lemeshow test (χ2 = 8.808, p = 0.359) illustrated a strong concordance between the model's predictions of poor prognosis and the actual incidence rate. CONCLUSION: CPVS has been shown to be effective in improving upper limb motor function in patients with cerebral palsy. Independent prognostic factors identified encompass motor function, age, and serum albumin levels. The composite predictive model shows potential for clinical applications.

Development and validation of a predictive model for poor prognosis of communication disorders in children with cerebral palsy after cervical perivascular sympathectomy.

Cervical perivascular sympathectomy (CPVS) can improve communication disorders in children with cerebral palsy (CP); however, there are no research reports on the factors affecting surgical efficacy. This study aimed to establish a nomogram for poor prognosis after CPVS. We collected data from 313 CP patients who underwent CPVS at the Neurosurgery Cerebral Palsy Center of the Second Affiliated Hospital of Xinjiang Medical University from January 2019 to January 2023. Among them, 70% (n = 216) formed the training cohort and 30% (n = 97) the validation cohort. The general data and laboratory examination data of both groups were analyzed. In training cohort, 82 (37.96%) showed improved postoperative communication function. Logistic analysis identified motor function, serum alkaline phosphatase, serum albumin, and prothrombin activity as the prognostic factors. Using these four factors, a prediction model was constructed with an area under the curve (AUC) of 0.807 (95% confidence interval [CI], 0.743-0.870), indicating its ability to predict adverse outcomes after CPVS. The validation cohort results showed an AUC of 0.76 (95% CI, 0.650-0.869). The consistency curve and Hosmer-Lemeshow test (χ2 = 10.988 and p = 0.202, respectively) demonstrated good consistency between the model-predicted incidence and the actual incidence of poor prognosis. Motor function, serum alkaline phosphatase, serum albumin, and prothrombin activity are independent risk factors associated with the prognosis of communication disorders after CPVS. The combined prediction model has a good clinical prediction effect and has promising potential to be used for early prediction of prognosis of CPVS.

Preparation of Resistant Starch Types III + V with Moderate Amylopullulanase and Its Effects on Bread Properties.

The potential of PulY103A (a moderate amylopullulanase originating from Bacillus megaterium) for resistant starch production under moderate conditions (40 °C; a pH of 6.5) was investigated. PulY103A was much more suitable for pea resistant starch production with a high growth rate of 3.63. The pea resistant starch (PSpa) produced with PulY103A had lower levels of swelling power and solubility and a better level of thermostability than native pea starch (PSn) and autoclaved PS (PSa). The starch crystallinity pattern was B + V, which indicated that the PSpa belonged to RS types III + V. In addition, PSpa was used for breadmaking. The results showed that the bread quality was not significantly influenced compared to the control group when the content of PSpa was under 10% (p > 0.05). The bread supplemented with 10% PSpa had a significantly increased TDF content compared to that of the control (p < 0.05). Moreover, the in vitro mineral bioavailability of the bread sample was influenced gently compared to other dietary fibers, and the bread sample changed from a high-glycemic-index (GI) food to a medium-GI food corresponding to white bread at the same concentration of PSpa. These results indicated that PSpa is a good candidate for the production of dietary foods.

N-Acetyltransferase 10 represses Uqcr11 and Uqcrb independently of ac4C modification to promote heart regeneration.

Translational control is crucial for protein production in various biological contexts. Here, we use Ribo-seq and RNA-seq to show that genes related to oxidative phosphorylation are translationally downregulated during heart regeneration. We find that Nat10 regulates the expression of Uqcr11 and Uqcrb mRNAs in mouse and human cardiomyocytes. In mice, overexpression of Nat10 in cardiomyocytes promotes cardiac regeneration and improves cardiac function after injury. Conversely, treating neonatal mice with Remodelin-a Nat10 pharmacological inhibitor-or genetically removing Nat10 from their cardiomyocytes both inhibit heart regeneration. Mechanistically, Nat10 suppresses the expression of Uqcr11 and Uqcrb independently of its ac4C enzyme activity. This suppression weakens mitochondrial respiration and enhances the glycolytic capacity of the cardiomyocytes, leading to metabolic reprogramming. We also observe that the expression of Nat10 is downregulated in the cardiomyocytes of P7 male pig hearts compared to P1 controls. The levels of Nat10 are also lower in female human failing hearts than non-failing hearts. We further identify the specific binding regions of Nat10, and validate the pro-proliferative effects of Nat10 in cardiomyocytes derived from human embryonic stem cells. Our findings indicate that Nat10 is an epigenetic regulator during heart regeneration and could potentially become a clinical target.

Efficacy and influencing factors of cervical perivascular sympathectomy in children with cerebral palsy.

BACKGROUND: There is a lack of research to determine the efficacy of cervical perivascular sympathectomy (CPVS) in children with cerebral palsy (CP). OBJECTIVE: This study aimed to evaluate the efficacy of CPVS in children with CP and analyze the associated influential factors. METHODS: Using the method of retrospective cohort studies, children who underwent CPVS were included in the CPVS group, whereas those who underwent selective posterior rhizotomy (SPR) were included in the SPR group. The Communication Function Classification System (CFCS) and Teacher Drooling Scale (TDS) were used to evaluate the communication function and salivation in the two groups before and 12 months after surgery and compare the surgical efficiency between the two groups, and the factors affecting the efficacy were screened by binary logistic regression. RESULTS: The study included 406 patients, 202 in the CPVS group and 204 in the SPR group. No significant differences were observed in the baseline characteristics (p > 0.05). The surgical efficacy of the CPVS group (47.01%) was significantly higher than that in the SPR group (9.81%) (χ2 = 71.08, p < 0.001). Binary logic regression analysis showed that preterm birth and Gross Motor Function Classification System (GMFCS) grade were influencing factors of surgical efficacy. Eighteen patients developed postoperative complications. CONCLUSION: CPVS is a safe and effective surgery for cerebral palsy. Preterm birth and GMFCS grade are independent factors affecting the efficacy of surgery.

An ultrahigh sensitivity acoustic sensor system for weak signal detection based on an ultrahigh-Q CaF2 resonator.

Acoustic sensors with ultrahigh sensitivity, broadband response, and high resolution are essential for high-precision nondestructive weak signal detection technology. In this paper, based on the size effect of an ultrahigh-quality (Q) calcium fluoride (CaF2) resonator, a weak acoustic signal is detected by the dispersive response regime in which an acoustic, elastic wave modulates the geometry and is converted to a resonance frequency shift. Through the structural design of the resonator, the sensitivity reaches 11.54 V/Pa at 10 kHz in the experiment. To our knowledge, the result is higher than that of other optical resonator acoustic sensors. We further detected a weak signal as low as 9.4 µPa/Hz1/2, which greatly improved the detection resolution. With a good directionality of 36.4 dB and a broadband frequency response range of 20 Hz-20 kHz, the CaF2 resonator acoustic sensing system can not only acquire and reconstruct speech signals over a long distance but also accurately identify and separate multiple voices in noisy environments. This system shows high performance in weak sound detection, sound source localization, sleep monitoring, and many other voice interaction applications.

Development of Bivalent mRNA Vaccines against SARS-CoV-2 Variants.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected billions of individuals and is the cause of the current global coronavirus disease 2019 (COVID-19) pandemic. We previously developed an mRNA vaccine (LVRNA009) based on the S protein of the Wuhan-Hu-1 strain; the phases I and II clinical trials showed that LVRNA009 has a promising safety and immunogenicity profile. In order to counteract the immune escape by SARS-CoV-2 variants of concern, a panel of mRNA vaccines was developed based on the S proteins of the Wuhan-Hu-1, Delta, Omicron BA.1, BA.2, and BA.5 strains, and each vaccine's protective potency against the virus variants was evaluated. Furthermore, to achieve excellent neutralization against SARS-CoV-2 variants, bivalent vaccines were developed and tested against the variants. We found that the monovalent Wuhan-Hu-1 or the Delta vaccines could induce high level of neutralization antibody and protect animals from the infection of the SARS-CoV-2 Wuhan-Hu-1 or Delta strains, respectively. However, serum samples from mice immunized with monovalent Delta vaccine showed relatively low virus neutralization titers (VNTs) against the pseudotyped virus of the Omicron strains. Serum samples from mice immunized with bivalent Delta/BA.1 vaccine had high VNTs against the pseudotyped Wuhan-Hu-1, Delta, and BA.1 strains but low VNTs against BA.2 and BA.5 (p < 0.05). Serum samples from mice immunized with Delta/BA.2 vaccine had high VNTs against the pseudotyped Wuhan-Hu-1, Delta, BA.1 and BA.2 strains but low VNTs against BA.5. Finally, serum samples from mice immunized with Delta/BA.5 vaccine had high VNTs against all the tested pseudotyped SARS-CoV-2 strains including the Wuhan-Hu-1, Delta, and Omicron variants (p > 0.05). Therefore, a bivalent mRNA vaccine with Delta/BA.5 combination is promising to provide broad spectrum immunity against all VOCs.

An mRNA-based rabies vaccine induces strong protective immune responses in mice and dogs.

Rabies is a lethal zoonotic disease that is mainly caused by the rabies virus (RABV). Although effective vaccines have long existed, current vaccines take both time and cost to produce. Messenger RNA (mRNA) technology is an emergent vaccine platform that supports rapid vaccine development on a large scale. Here, an optimized mRNA vaccine construct (LVRNA001) expressing rabies virus glycoprotein (RABV-G) was developed in vitro and then evaluated in vivo for its immunogenicity and protective capacity in mice and dogs. LVRNA001 induced neutralizing antibody production and a strong Th1 cellular immune response in mice. In both mice and dogs, LVRNA001 provided protection against challenge with 50-fold lethal dose 50 (LD50) of RABV. With regards to protective efficiency, an extended dosing interval (14 days) induced greater antibody production than 3- or 7-day intervals in mice. Finally, post-exposure immunization against RABV was performed to evaluate the survival rates of dogs receiving two 25 µg doses of LVRNA001 vs. five doses of inactivated vaccine over the course of three months. Survival rate in the LVRNA001 group was 100%, whereas survival rate in the inactivated vaccine control group was only 33.33%. In conclusion, these results demonstrated that LVRNA001 induced strong protective immune responses in mice and dogs, which provides a new and promising prophylactic strategy for rabies.

Large-scale flexible-resonators with temperature insensitivity employing superoleophobic substrates.

Whispering gallery mode polymer resonators are becoming competitive with devices made of other materials, however, the inherent thermal sensitivity of the materials and the small size limit their applications, such as high-precision optical gyroscope. Here, a method is proposed for fabricating large-scale NOA65 resonators with quality factors greater than 105 on a chip employing superoleophobic. The sandwich structure as the core layer of resonator is used to present the flexible remodeling characteristics, the surface roughness remains below 1 nm when the diameter changes by more than 25%. Importantly, theoretical and experimental results show that under the tuning action of external pressure, the equivalent thermal expansion coefficient of the resonator gradually approaches the glass sheet on both sides with the variation of 2 × 10-4 /°C∼0.9 × 10-4 /°C, and the corresponding temperature response range of 0.12 nm/°C∼-0.056 nm/°C shows the promise of temperature insensitivity resonators on a chip.

Causal association between metro transits and air quality: China's evidence.

In policy remedies, transportation infrastructure such as metro transit is widely considered to be an important and effective means to reduce air pollution. However, the policy prediction that metro transits reduce air pollution depends on driver responses. China provides an appropriate context to explore the answer since its major cities have expanded their metro transit systems in recent years, which enables us to exploit a natural experiment. Accordingly, a sharp regression discontinuity is employed to evaluate the impact of 112 metro lines (with an accumulation of 3286 km) on air quality. Evidence shows that the opening of metro transits has a negative causal effect on air pollution. The results are robust to several alternative specifications. Furthermore, a heterogeneity analysis demonstrates that metro openings have a time-varying impact on air pollution, which is larger during rush hours. We anticipate that the air quality improvement in China caused by metro transits can generate large welfare gains.

miR­125a­5p reverses epithelial­mesenchymal transition and restores drug sensitivity by negatively regulating TAFAZZIN signaling in breast cancer.

MicroRNA (miR)­125a­5p represses tafazzin phospholipid­lysophospholipid transacylases (TAFAZZIN) expression and inhibits the epithelial­mesenchymal transition (EMT) of ovarian cancer cells. EMT was found to have a crucial role in the acquisition of chemoresistance. Thus, the present study aimed to determine whether miR­125a­5p reverses EMT and restores drug sensitivity by negatively regulating TAFAZZIN in breast cancer. The expression of miR­125a­5p/TAFAZZIN and its association with chemotherapy response were determined in tissue samples from patients with breast cancer. Furthermore, the effects of miR­125a­5p on breast cancer cells were elucidated using cell proliferation and cell apoptosis assays. Then, the regulatory mechanism of miR­125a­5p in breast cancer was investigated by reverse transcription­quantitative PCR, western blotting, dual­luciferase reporter and RNA immunoprecipitation assays. The results demonstrated that miR­125a­5p inhibited the EMT of MCF­7/adriamycin (Adr) breast cancer cells, as well as decreased the proliferation and increased the apoptosis of breast cancer cells treated with Adr/docetaxel. In addition, miR­125a­5p downregulated the expression levels of TAFAZZIN, Transglutaminase 2, phosphorylated­AKT, N­cadherin, vimentin and proliferating cell nuclear antigen, and significantly increased those of E­cadherin, cleaved caspase-3 and Bax in MCF7/Adr cells. Similar results were obtained with small interfering RNA­TAFAZZIN. Moreover, TAFAZZIN was identified as a direct target of miR­125a­5p in MCF7/Adr breast cancer cells. In addition, increased miR­125a­5p expression was observed in breast tumors from patients exhibiting a chemotherapy response, and TAFAZZIN mRNA expression was elevated in patients with no chemotherapy response. Hence, miR­125a­5p expression was negatively correlated with TAFAZZIN mRNA expression in breast cancer tissues. All these data suggested that miR­125a­5p reverses EMT and restores drug sensitivity by negatively regulating TAFAZZIN in breast cancer and, therefore, has potential as a novel therapeutic target for this disease.

Immunization with a recombinant fusion of porcine reproductive and respiratory syndrome virus modified GP5 and ferritin elicits enhanced protective immunity in pigs.

Porcine reproductive and respiratory syndrome (PRRS) has caused huge economic losses in the swine industry worldwide. Live and inactivated vaccines have only been partially successful in generating protective immune responses. The PRRS virus (PRRSV) glycoprotein 5 (GP5) is a major viral antigenic target and is thus suitable for development of genetically engineered PRRSV vaccines. Here, a modified GP5 and ferritin were fused and expressed using a baculovirus system to generate a GP5m-ferritin nanoparticle vaccine. We demonstrated that the GP5m-ferritin vaccine elicited higher serum antibody titers in pigs than inactivated PRRSV. Moreover, immunization with GP5m-Ft promoted a Th1-dominant cellular immune response and enhanced specific T-lymphocyte immune responses. GP5m-ferritin-vaccinated pigs had significantly lower mean rectal temperatures, respiratory scores, viremia, and macroscopic and microscopic lung lesion scores post-challenge compared with unvaccinated pigs. These results indicated that GP5m-ferritin subunit vaccines can elicit specific protective immune responses and represent promising vaccine candidates.

The dynamic linkage among urbanisation, industrialisation and carbon emissions in China: Insights from spatiotemporal effect.

Understanding how urbanisation and industrialisation interact with carbon emissions (CEs) among different regions informs the approaches for achieving emissions reduction targets and promoting regional economic development. However, this topic has not been adequately addressed in previous studies. Therefore, it is of great significance to conduct an empirical investigation of the interactions between urbanisation, industrialisation and CEs. Aided by the gravity model, the barycentre trajectories of urbanisation, industrialisation and CEs are analysed. Additionally, the total transfer amounts of industry and CEs are quantitatively examined by using an improved shift-share analysis method, and the spatial aggregation features of urbanisation, industrialisation and CEs are elucidated by constructing a three-dimensional (3D) contribution matrix model. The key conclusions are as follows: (1) The barycentre of industrialisation and urbanisation migrated to the southwest, while that of CE migrated to the northwest and the speed of this movement was faster than that of urbanisation and industrialisation. (2) Industrial transfer mainly occurred in 2005-2012, while the turning point of industrial upgrading appeared in 2013. Furthermore, midwestern regions mainly underwent industrial transfer, while northwest regions always showed CEs transfer from 2005 to 2016. (3) To achieve coordination among urbanisation, industrialisation and emission reductions, 30 provinces are grouped into 8 types according to the 3D contribution matrix model, and optimisation strategies are proposed to highlight regional disparities. These findings have significant implications for making informed decisions regarding urbanisation and industrialisation development as well as emissions-reduction policies.

Efficient mucosal vaccination of a novel classical swine fever virus E2-Fc fusion protein mediated by neonatal Fc receptor.

Classical swine fever (CSF) remains one of the most important highly contagious and fatal viral disease of swine with high morbidity and mortality. CSF is caused by classical swine fever virus (CSFV), a small, enveloped RNA virus of the genus Pestivirus. The aim of this study was to construct the a novel CSFV Fc-fusion recombinant protein and evaluate the efficacy as a vaccine against CSFV. Here, we obtained a novel subunit vaccine expressing CSFV E2 recombinant fusion protein in CHO-S cells. Functional analysis revealed that CSFV Fc-fusion recombinant protein (CSFV-E2-Fc) could bind to FcγRI on antigen-presenting cells (APCs) and significantly increase IgA levels in serum and feces, inducing stronger mucosal immune response in swine. Additionally, CSFV-E2-Fc immunization enhanced CSFV-specific T cell immune response with a Th1-like pattern of cytokine secretion, remarkably stimulated the Th1-biased cellular immune response and humoral immune response. Further, the protective effects of CSFV-E2-Fc subunit vaccines were confirmed. The data suggest that CSFV E2-Fc recombinant fusion protein may be a promising candidate subunit vaccine to elicit immune response and protect against CSFV.

Comparison of gE/gI- and TK/gE/gI-Gene-Deleted Pseudorabies Virus Vaccines Mediated by CRISPR/Cas9 and Cre/Lox Systems.

Pseudorabies (PR), caused by pseudorabies virus (PRV), is an acute and febrile infectious disease in swine. To eradicate PR, a more efficacious vaccine needs to be developed. Here, the gE/gI- and TK/gE/gI-gene-deleted recombinant PRV (rGXΔgE/gI and rGXΔTK/gE/gI) are constructed through CRISPR/Cas9 and Cre/Lox systems. We found that the rGXΔTK/gE/gI was safer than rGXΔgE/gI in mice. Additionally, the effects of rGXΔgE/gI and rGXΔTK/gE/gI were further evaluated in swine. The rGXΔgE/gI and rGXΔTK/gE/gI significantly increased numbers of IFN-γ-producing CD4+ and CD8+ T-cells in swine, whereas there was no difference between rGXΔgE/gI and rGXΔTK/gE/gI. Moreover, rGXΔgE/gI and rGXΔTK/gE/gI promoted a PRV-specific humoral immune response. The PRV-specific humoral immune response induced by rGXΔgE/gI was consistent with that caused by rGXΔTK/gE/gI. After the challenge, swine vaccinated with rGXΔgE/gI and rGXΔTK/gE/gI showed no clinical signs and viral shedding. However, histopathological detection revealed that rGXΔgE/gI, not rGXΔTK/gE/gI, caused pathological lesions in brain and lung tissues. In summary, these results demonstrate that the TK/gE/gI-gene-deleted recombinant PRV was safer compared with rGXΔgE/gI in swine. The data imply that the TK/gE/gI-gene-deleted recombinant PRV may be a more efficacious vaccine candidate for the prevention of PR.