Severe Acute Respiratory Syndrome(SARS)is a deadly infectious disease caused by SARS Coronavirus(SARS-CoV).Inactivated SARS-CoV has been explored as a vaccine against SARS-CoV.However,safe and potent adjuvants,especially with more efficient and economical needle-free vaccination are always needed more urgently in a pandemic.The development of a safe and effective mucosal adjuvant and vaccine for prevention of emergent infectious diseases such as SARS will be an important advancement.PIKA,a stabilized derivative of Poly(I:C),was previously reported to be safe and potent as adjuvant in mouse models.In the present study,we demonstrated that the intraperitoneal and intranasal co-administration of inactivated SARS-CoV vaccine together with this improved Poly(I:C)derivative induced strong anti-SARS-CoV mucosal and systemic humoral immune responses with neutralizing activity against pseudotyped virus.Although intraperitoneal immunization of inactivated SARS-CoV vaccine alone could induce a certain level of neutralizing activity in serum as well as in mucosal sites,co-administration of inactivated SARS-CoV vaccine with PIKA as adjuvant could induce a much higher neutralizing activity.When intranasal immunization was used,PIKA was obligatorily for inducing neutralizing activity in serum as well as in mucosal sites and was correlated with both mucosal IgA and mucosal IgG response.Overall,PIKA could be a good mucosal adjuvant candidate for inactivated SARS-CoV vaccine for use in possible future pandemic.

Objective To investigate the clinical manifestations,imaging features,molecular genetic characteristics and possible pathogenic mechanisms of hereditary cerebral small vessel disease (CSVD) caused by heterozygous mutation of HtrA serine protease-1 (HTRA1) gene.Methods The clinical data of a Chinese Han family with CSVD carrying a heterozygous mutation of HTRA 1 gene,which came from the Department of Neurology,Henan Provincial People's Hospital in March 2018,were analyzed retrospectively.The clinical and radiographic features were summarized.Several high-throughput whole exon high-throughput sequencing was used to capture the mutation sites and the Sanger sequencing was used to validate the results.The family diagram was drawn and the 3D model construction and mutation function prediction were performed using silico tools.The relevant literature was reviewed and the pathogenesis was explored.Results The pedigree map showed that the family had an autosomal dominant inheritance pattern.Three generations of the family were investigated,and three family members in the same generation suffered from the disease.The first symptom of the proband was diplopia at the age of 39,accompanied by recurrent stroke,cognitive impairment and mood disorders,without alopecia.Head magnetic resonance imaging revealed bilateral diffuse,symmetric lesions,multiple lacunar infarcts,perivascular space,and microbleeds.The elder sister of the proband developed symptoms of left limb weakness at the age of 46,whose other clinical and imaging features were similar to those of the proband.The proband's mother died at the age of 59 due to repeated strokes.Whole exon sequencing indicated heterozygous missense mutation at c.821G＞A locus of HTRA1 gene in the proband and her 4th elder sibling,which was a new pathogenic mutation after consulting several mutation sites of databases.Function prediction suggested pathogenicity.Conclusions The heterozygous mutation of c.821G＞A in HTRA1 gene may lead to autosomal dominant CVSD.This genetic type should be given clinical attention.

Objective:To summarize the clinical and imaging features of five patients of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) with cysteine-sparing NOTCH3 gene missense mutations and explore potential pathogenicity of gene mutations.Methods:The clinical data from five patients who were admitted to the People′s Hospital of Zhengzhou University from March 2017 to November 2018 were collected. The patients were found to carry cysteine-sparing NOTCH3 gene mutations through genetic testing and diagnosed pathologically. They were probands confirmed from five unrelated family and all five patients were performed full exon detection and skin biopsy.Results:Genetic testing identified five patients with cysteine-sparing NOTCH3 gene missense mutations, a total of five different mutations, including p.R75Q, p.D80G, p.V237M, p.S1418L and p.R1761H. The first three mutations were found in the epidermal growth factor-like repeats (EGFr), the latter two mutations near the transmembrane domain. Granular osmiophilic material was identified in all cases examined with skin biopsy. The age at initial symptom onset of these five cases was ranged from 22 to 58 years and three cases presented cardiovascular risk factors. The primary clinical manifestations included migraine in one case, ischemic stroke in three cases, psychiatric disturbances in four cases, cognitive dysfunction in five cases, while gait disturbance, pseudobulbar palsy, and seizures accounted for only one case each. Magnetic resonance imaging of five patients all showed white matter hyperintensities (WMLs) and lacunar infarcts, and WMLs involved the anterior temporal pole and external capsules in three cases separately. According to the criteria proposed by Mui?o et al for evaluating the pathogenicity of cysteine-sparing NOTCH3 mutations, all five mutations are potentially pathogenic.Conclusions:Most characteristics of CADASIL patients with cysteine-sparing NOTCH3 gene mutations are similar to those of CADASIL patients with cysteine NOTCH3 gene mutations. Mutations not involving the EGFr may also have potential pathogenicity, and the specific mechanism still needs further study.

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV). RVF is mainly prevalent on the Arabian Peninsula, the African continent, and several islands in the Indian Ocean near southeast Africa. RVFV has been classified by the World Organisation for Animal Health (OIE) as a category A pathogen. To avoid biological safety concerns associated with use of the pathogen in RVFV neutralization assays, the present study investigated and established an RVFV pseudovirus-based neutralization assay. This study used the human immunodeficiency virus (HIV) lentiviral packaging system and RVFV structural proteins to successfully construct RVFV pseudoviruses. Electron microscopy observation and western blotting indicated that the size, structure, and shape of the packaged pseudoviruses were notably similar to those of HIV lentiviral vectors. Infection inhibition assay results showed that an antibody against RVFV inhibited the infective ability of the RVFV pseudoviruses, and an antibody neutralization assay for RVFV detection was then established. This study has successfully established a neutralization assay based on RVFV pseudoviruses and demonstrated that this method can be used to effectively evaluate antibody neutralization.
Africa ; Animals ; Blotting, Western ; HIV ; Indian Ocean ; Islands ; Methods ; Microscopy, Electron ; Product Packaging ; Rift Valley fever virus ; Rift Valley Fever ; Zoonoses

Objective:To explore the diagnostic value of 68Ga-fibroblast activation protein inhibitor (FAPI) PET for the restaging of patients with colorectal cancer and its impact on treatment strategy. Methods:Patients with colorectal cancer who underwent 68Ga-FAPI PET imaging in the PET Center of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June 2020 to June 2021 were retrospectively analyzed. PET images were evaluated by 3 experienced imaging physicians. Biopsy or postoperative pathology, clinical and imaging follow-up results were as the gold standard. The diagnostic value of PET was compared with conventional imaging (CT/MR), and the impact of 68Ga-FAPI PET on guiding treatment was evaluated. χ2 test and Fisher exact test were used to compare the differences between groups. Results:A total of 33 patients were included (17 males, 16 females, age (52.8±12.3) years), of which 24 were finally diagnosed as recurrence/metastases/progression. The accuracy, sensitivity, specificity, positive predictive value and negative predictive value of 68Ga-FAPI PET in detecting recurrence/metastases/progression of colorectal cancer were 93.9%(31/33), 100%(24/24), 7/9, 92.3%(24/26) and 7/7, respectively. Its accuracy, sensitivity and negative predictive value were significantly higher than those of conventional imaging (64.5%(20/31), 56.5%(13/23) and 7/17; χ2 values: 8.549 and 10.786, all P<0.05). Compared with the clinical or pathological stage before examination, 68Ga-FAPI PET led upstaging to stage Ⅳ in 12 patients (50.0%, 12/24). Of the 31 patients who were correctly diagnosed by 68Ga-FAPI PET, the treatment regimen of 22 patients (71.0%) was changed because of 68Ga-FAPI PET imaging. Conclusion:68Ga-FAPI PET has good diagnostic performance in the restaging of colorectal cancer, which is helpful to further guide clinical treatment strategy.

Objective:To investigate the diagnostic and prognostic value of 68Ga-fibroblast activation protein inhibitor (FAPI) PET for hepatobiliary malignancies. Methods:From July 2020 to February 2023, 33 patients (23 males, 10 females; age (55.4±13.5) years) with suspected or confirmed liver or biliary tract malignancies who underwent 68Ga-FAPI PET in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology were retrospectively analyzed. PET images were evaluated by 3 experienced nuclear medicine physicians, and the results of biopsy or postoperative pathology, clinical and imaging follow-up were used as the gold standard. One-way analysis of variance and least significant difference t test were used to compare the differences among groups. Survival analysis was performed using Kaplan-Meier curves and the log-rank test. Results:Of 33 patients, 14 performed PET for initial diagnosis and staging, and 19 for restaging. There were 14 patients with hepatocellular carcinoma (HCC), 13 patients with cholangiocarcinoma (CCA), and 6 patients with gallbladder carcinoma (GBC). The primary tumor of HCC, CCA and GBC all showed significant 68Ga-FAPI uptake, with no statistically significant difference in SUV max among groups ( F=1.58, P=0.250). The sensitivities of 68Ga-FAPI PET for initial diagnosis and restaging of hepatobiliary malignancies were 14/14 and 15/15, respectively. Compared with conventional imaging, 68Ga-FAPI PET changed the diagnosis and staging in 29.2%(7/24) patients. The treatment strategy was changed in 30.3%(10/33) patients with malignant tumors due to 68Ga-FAPI PET findings. Follow-up showed 22 cases survived and 11 cases died, with the overall survival of 355.56(80.00, 516.97) d, and 1- and 2-year survival rates were 68.2% and 57.9%, respectively. Semi-quantitative 68Ga-FAPI PET parameters such as SUV max, target-liver ratio (TLR), and target-blood ratio (TBR) had no significant prognostic value, but the prognosis of the group without distant metastases diagnosed by 68Ga-FAPI PET was significantly better than that of the group with distant metastasis ( P=0.032). Conclusion:68Ga-FAPI PET has high sensitivity for the diagnosis of hepatobiliary malignancies, which can help guide treatment decisions and prognosis evaluation.

Objective:To explore the feasibility of statistical parametric mapping (SPM) aided semi-quantitative analysis in 11C-Pittsburgh compound B (PIB) β-amyloid (Aβ) PET imaging acquired by hybrid PET/MR, and evaluate its possibility in assisting the diagnosis or differential diagnosis for cognitive impairment. Methods:From January 2018 to September 2019, 13 Alzheimer′s disease (AD) patients (4 males, 9 females; age (59.2±5.8) years) and 10 vascular cognitive disorders (VCD) patients (9 males, 1 female; age (59.5±11.5) years) who underwent 11C-PIB PET/MR in PET center of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology were retrospectively analyzed. The standardized uptake value ratio (SUVR) of eight key brain regions (cerebral white matter, striatum, thalamus, posterior cingulate gyrus, frontal cortex, posterior parietal cortex, lateral temporal cortex and occipital cortex) to cerebellum cortex were obtained by manual delineation and SPM-aided semi-automatic segmentation with the help of synchronous three-dimensional T 1 weighted imaging (3D T 1WI). Pearson correlation analysis was carried out on the SUVR obtained by the two methods. Independent-sample t test and paired t test were used to analyze the data. Results:There was no significant difference between AD group and VCD group in age and Mini-Mental State Examination (MMSE) score (19.7±4.7 vs 21.7±3.8; t values: 0.095 and 1.098, both P>0.05). Except thalamus( r=0.179, P=0.413), there were good correlations between SUVR obtained by segmentation and delineation in the other 7 key regions ( r values: 0.678-0.893, all P<0.05). The SUVR of 8 key regions obtained by the two methods in AD group was significantly higher than that in VCD group (1.519-2.055 vs 1.105-1.618; t values: 2.799-11.582, all P<0.01). The SUVR of striatum (1.942±0.205), posterior cingulate gyrus (1.915±0.249), frontal lobe (1.983±0.264), parietal lobe (2.008±0.296) and temporal cortex (1.931±0.254) in AD group was significantly higher than that of cerebral white matter (1.746±0.192; t values: 3.793-6.992, all P<0.01). But in VCD group, there was no region with the SUVR higher than that of cerebral white matter. Conclusions:Hybrid PET/MR can acquire the PET and MRI images synchronously, which can realize the accurate brain segmentation and obtain the semi-quantitative data of key brain regions aided by SPM. The method can analyze the characteristics and differences of amyloid imaging in AD and VCD, which is expected to provide an accurate imaging analysis method for the diagnosis and differential diagnosis of cognitive disorders.

School-age children are in a specific development stage corresponding to juvenility, when the white matter of the brain experiences ongoing maturation. Diffusion-weighted magnetic resonance imaging (DWI), especially diffusion tensor imaging (DTI), is extensively used to characterize the maturation by assessing white matter properties in vivo. In the analysis of DWI data, spatial normalization is crucial for conducting inter-subject analyses or linking the individual space with the reference space. Using tensor-based registration with an appropriate diffusion tensor template presents high accuracy regarding spatial normalization. However, there is a lack of a standardized diffusion tensor template dedicated to school-age children with ongoing brain development. Here, we established the school-age children diffusion tensor (SACT) template by optimizing tensor reorientation on high-quality DTI data from a large sample of cognitively normal participants aged 6-12 years. With an age-balanced design, the SACT template represented the entire age range well by showing high similarity to the age-specific templates. Compared with the tensor template of adults, the SACT template revealed significantly higher spatial normalization accuracy and inter-subject coherence upon evaluation of subjects in two different datasets of school-age children. A practical application regarding the age associations with the normalized DTI-derived data was conducted to further compare the SACT template and the adult template. Although similar spatial patterns were found, the SACT template showed significant effects on the distributions of the statistical results, which may be related to the performance of spatial normalization. Looking forward, the SACT template could contribute to future studies of white matter development in both healthy and clinical populations. The SACT template is publicly available now ( https://figshare.com/articles/dataset/SACT_template/14071283 ).

BACKGROUND: Liver cancer is largely resistant to chemotherapy. This study aimed to identify the effective chemotherapeutics for β-catenin-activated liver cancer which is caused by gain-of-function mutation of catenin beta 1 ( CTNNB1 ), the most frequently altered proto-oncogene in hepatic neoplasms. METHODS: Constitutive β-catenin-activated mouse embryonic fibroblasts (MEFs) were established by deleting exon 3 ( β-catenin Δ(ex3)/+ ), the most common mutation site in CTNNB1 gene. A screening of 12 widely used chemotherapy drugs was conducted for the ones that selectively inhibited β-catenin Δ(ex3)/+ but not for wild-type MEFs. Untargeted metabolomics was carried out to examine the alterations of metabolites in nucleotide synthesis. The efficacy and selectivity of methotrexate (MTX) on β-catenin-activated human liver cancer cells were determined in vitro . Immuno-deficient nude mice subcutaneously inoculated with β-catenin wild-type or mutant liver cancer cells and hepatitis B virus ( HBV ); β-catenin lox(ex3)/+ mice were used, respectively, to evaluate the efficacy of MTX in the treatment of β-catenin mutant liver cancer. RESULTS: MTX was identified and validated as a preferential agent against the proliferation and tumor formation of β-catenin-activated cells. Boosted nucleotide synthesis was the major metabolic aberration in β-catenin-active cells, and this alteration was also the target of MTX. Moreover, MTX abrogated hepatocarcinogenesis of HBV ; β-catenin lox(ex3)/+ mice, which stimulated concurrent Ctnnb1- activated mutation and HBV infection in liver cancer. CONCLUSION MTX is a promising chemotherapeutic agent for β-catenin hyperactive liver cancer. Since repurposing MTX has the advantages of lower risk, shorter timelines, and less investment in drug discovery and development, a clinical trial is warranted to test its efficacy in the treatment of β-catenin mutant liver cancer.
Mice ; Animals ; Humans ; Methotrexate/therapeutic use* ; Mice, Nude ; beta Catenin/metabolism* ; Fibroblasts/metabolism* ; Liver Neoplasms/metabolism* ; Hepatitis B virus ; Nucleotides