BACKGROUND: Epoxyeicosatrienoic acids (EETs), which are metabolites of arachidonic acid catalyzed by cytochrome P450 epoxygenase, are degraded into inactive dihydroxyeicosatrienoic acids by soluble epoxide hydrolase (sEH). Many studies have revealed that sEH gene deletion exerts protective effects against diabetes. Vascular calcification is a common complication of diabetes, but the potential effects of sEH on diabetic vascular calcification are still unknown. METHODS: The level of aortic calcification in wild-type and Ephx2-/- C57BL/6 diabetic mice induced with streptozotocin was evaluated by measuring the aortic calcium content through alizarin red staining, immunohistochemistry staining, and immunofluorescence staining. Mouse vascular smooth muscle cell lines (MOVAS cells) treated with β-glycerol phosphate (0.01 mol/L) plus advanced glycation end products (50 mg/L) were used to investigate the effects of sEH inhibitors or sEH knockdown and EETs on the calcification of vascular smooth muscle cells, which was detected by Western blotting, alizarin red staining, and Von Kossa staining. RESULTS: sEH gene deletion significantly inhibited diabetic vascular calcification by increasing levels of EETs in the aortas of mice. EETs (especially 11,12-EET and 14,15-EET) efficiently prevented the osteogenic transdifferentiation of MOVAS cells by decreasing nidogen-2 (NID2) expression. Interestingly, suppressing sEH activity by small interfering ribonucleic acid or specific inhibitors did not block osteogenic transdifferentiation of MOVAS cells induced by β-glycerol phosphate and advanced glycation end products. NID2 overexpression significantly abolished the inhibitory effect of sEH gene deletion on diabetic vascular calcification. Moreover, NID2 overexpression mediated by adeno-associated virus 9 vectors markedly increased insulin-like growth factor 2 (IGF2) and phospho-ERK1/2 expression in MOVAS cells. Overall, sEH gene knockout inhibited diabetic vascular calcification by decreasing aortic NID2 expression and, then, inactivating the downstream IGF2-ERK1/2 signaling pathway. CONCLUSIONS sEH gene deletion markedly inhibited diabetic vascular calcification through repressed osteogenic transdifferentiation of vascular smooth muscle cells mediated by increased aortic EET levels, which was associated with decreased NID2 expression and inactivation of the downstream IGF2-ERK1/2 signaling pathway.
Animals ; Mice ; Vascular Calcification/metabolism* ; Mice, Inbred C57BL ; Epoxide Hydrolases/metabolism* ; Diabetes Mellitus, Experimental/genetics* ; Male ; Gene Deletion ; MAP Kinase Signaling System/genetics* ; Cell Line ; Immunohistochemistry ; Muscle, Smooth, Vascular/metabolism* ; Signal Transduction/genetics* ; Mice, Knockout

OBJECTIVES: We present a new low-dose CT reconstruction method using sub-pixel and anisotropic diffusion. METHODS: The sub-pixel intensity values and their second-order differences were obtained using linear interpolation techniques, and the new gradient information was then embedded into an anisotropic diffusion process, which was introduced into a penalty-weighted least squares model to reduce the noise in low-dose CT projection data. The high-quality CT image was finally reconstructed using the classical filtered back-projection (FBP) algorithm from the estimated data. RESULTS: In the Shepp-Logan phantom experiments, the structural similarity (SSIM) index of the CT image reconstructed by the proposed algorithm, as compared with FBP, PWLS-Gibbs and PWLS-TV algorithms, was increased by 28.13%, 5.49%, and 0.91%, the feature similarity (FSIM) index was increased by 21.08%, 1.78%, and 1.36%, and the root mean square error (RMSE) was reduced by 69.59%, 18.96%, and 3.90%, respectively. In the digital XCAT phantom experiments, the SSIM index of the CT image reconstructed by the proposed algorithm, as compared with FBP, PWLS-Gibbs and PWLS-TV algorithms, was increased by 14.24%, 1.43% and 7.89%, the FSIM index was increased by 9.61%, 1.78% and 5.66%, and the RMSE was reduced by 26.88%, 9.41% and 18.39%, respectively. In clinical experiments, the SSIM index of the image reconstructed using the proposed algorithm was increased by 19.24%, 15.63% and 3.68%, the FSIM index was increased by 4.30%, 2.92% and 0.43%, and the RMSE was reduced by 44.60%, 36.84% and 15.22% in comparison with FBP, PWLS-Gibbs and PWLS-TV algorithms, respectively. CONCLUSIONS The proposed method can effectively reduce the noises and artifacts while maintaining the structural details in low-dose CT images.
Tomography, X-Ray Computed/methods* ; Algorithms ; Phantoms, Imaging ; Anisotropy ; Image Processing, Computer-Assisted/methods* ; Humans ; Radiation Dosage

ObjectiveTo explore the potential molecular mechanism of Qizhu Kang'ai Formula （芪术抗癌方， QZKAF） for the treatment of colorectal cancer （CRC）. MethodsNetwork pharmacology was used to analyze the active ingredients and targets of QZKAF for CRC， and analyze the key targets of QZKAF for the treatment of CRC by gene function annotation （GO） and Kyoto Encyclopedia of Genomes （KEGG） pathway enrichment analysis. Molecular docking was applied to predict the binding activity of the core active ingredients to the key targets. A orthotopic transplantation tumor mice model of CRC was established to validate the key targets of QZKAF for CRC obtained from network pharmacology analysis. Forty-eight mice were randomly divided into the sham operation group， the model group， the 5-fluorouracil （5-Fu） group， and the QZKAF low-， medium-， and high-dose groups， with 8 mice in each group. Except for the sham operation group， the remaining groups underwent colon cancer orthotopic transplantation tumor modeling. The 5-Fu group was given 30 mg/kg of 5-Fu by intraperitoneal injection once every 3 days on the alternate day after modeling， while the QZKAF low-， medium-， and high-dose groups were given 2.925， 5.85， and 11.7 g/（kg·d） of QZKAF by gastric gavage， respectively， and the sham-operation group and the model group were gavaged with 0.1 ml/10 g of normal saline every day， all for 21 days. The in situ tumors mass and the number of liver metastases were compared between the groups. The pathological changes of colon tumor tissues were observed by HE staining， and the protein expression of protein tyrosine phosphatase nonreceptor type 1 （PTPN1）， vinculin， integrin subunit αν， integrin subunit β3， and E-cadherin were detected in colon tumor tissues by Western blot. ResultsNetwork pharmacology screening yielded that the top six core active ingredients of QZKAF intervening in CRC were quercetin， kaempferol， apigenin， luteolin， baicalein and ursolic acid. There were 212 targets of action， and the ranked top three were prostaglandin endoperoxide synthase 1 （PTGS1）， prostaglandin endoperoxide synthase 2 （PTGS2）， and PTPN1， which may be the key targets of QZKAF in the treatment of CRC. These key targets were significantly enriched mainly in phosphatidylinositol 3-kinase/protein kinase B （PI3K-Akt） signaling pathway， focal adhesion and adhesion junction. Molecular docking results： except for PTGS1 with better binding activity to quercetin， kaempferol， and apigenin （binding energy ≥-7.0 kcal/mol）， PTGS1 showed strong binding activity to lignans， baicalein， ursolic acid， as well as PTGS2 and PTPN1 to the six core active ingredients （binding energy ＜-7.0 kcal/mol）. Experimental validation results： the protein expression of PTPN1， vinculin， integrin subunit αν， integrin subunit β3 in the colon tumor tissues of mice in the model group significantly increased， and the expression of E-cadherin significantly decreased compared to those in the sham operation group （P＜0.05）. Compared to those in the model group， the mass of the in situ tumors was reduced， and the number of hepatic metastasis nodules decreased in the high- and medium-dose QZKAF groups （P＜0.05）； the expression levels of PTNP1， vinculin， integrin subunit αν， integrin subunit β3 and E-cadherin in all QZKAF groups and 5-Fu group showed different degrees of retracement， and the changes of the indicators in all QZKAF groups showed a certain degree of dose-dependence （P＜0.05）. HE staining showed that the nuclei of tumor cells in the model group were mostly schizophrenic， and there were different degrees of nuclear fragmentation of tumor cells in all QZKAF groups with more in the medium- and high-dose groups. ConclusionQZKAF could inhibit the growth of in situ tumors and liver metastasis of CRC. Its mechanism might be related to the regulation of tumor cell-cell and tumor cell-extracellular matrix adhesion by PTPN1.

Objective:To investigate the impact of total cerebral small vessel disease (CSVD) burden on cognitive function in patients with minor ischemic stroke.Methods:Patients with first-ever acute minor ischemic stroke (AMIS) admitted to the Department of Neurology, Shenzhen Luohu District People's Hospital from January 2021 to December 2023 were included prospectively. According to the total CSVD burden score, they were divided into 0-2-point group, 3-point group, and 4-point group. According to the Clinical Dementia Rating (CDR) score at 6 months after onset, they were divided into a cognitively normal group and a post-stroke cognitive impairment (PSCI) group. The cognitive function assessment was conducted at 6 months after onset. The Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used to assess the overall cognitive function. The Auditory Verbal Learning Test-Huashan version (AVLT-H) was used to assess memory, the Trail Making Test (TMT) was used to assess executive function and attention, the Boston Naming Test (BNT) was used to assess language ability, and the Clock Drawing Test (CDT) was used to assess visual spatial function. Multivariate logistic regression analysis was used to determine the independent influencing factors of PSCI. Results:A total of 111 patients with AMIS were enrolled, including 53 males (47.7%), aged 69.18±7.41 years. 77 cases (69.4%) in 0-2-point group, 18 cases (16.2%) in 3-point group, and 16 cases (14.4%) in 4-point group; 39 cases (35.1%) experienced PSCI. Univariate analysis showed that age, and the proportion of patients with hypertension, diabetes, smoking, total CSVD burden score of 3 and 4 in the PSCI group was significantly higher than those in the cognitive normal group (all P<0.05), and the years of education was significantly lower than that in the cognitive normal group ( P<0.05). Multivariate logistic regression analysis showed that after adjusting for confounding factors (age, years of education, hypertension, diabetes, smoking), the total CSVD burden score 3 (compared with 0-2 points: odds ratio [ OR] 16.627, 95% confidence interval [ CI] 3.548-77.925; P<0.001) and 4 (compared to 0-2 points: OR 4.435, 95% CI 1.786-11.011; P=0.001) were the independent risk factors for PSCI. There were significant differences in MMSE, MoCA, AVLT-H, TMT-A, TMT-B, CDT, and CDR scores among different total CSVD burden score groups at 6 months after onset (all P<0.05). The overall cognitive function and functional scores of various cognitive domains gradually decreased with the increase of total CSVD burden score in patients with AMIS at 6 months after onset. Conclusions:In patients with AMIS with higher total CSVD burden at 6 months after onset, their overall cognitive function, memory, executive function, attention, and visual spatial function are decreased, and the incidence of PSCI is significantly increased.

The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases, such as depression and anxiety. Meanwhile, the endocannabinoid system plays a crucial role in regulating emotions and mainly functions through the cannabinoid type-1 receptor (CB₁R), which is strongly expressed in the amygdala of non-human primates (NHPs). However, it remains largely unknown how the CB₁Rs in the amygdala of NHPs regulate mental diseases. Here, we investigated the role of CB₁R by knocking down the cannabinoid receptor 1 (CNR1) gene encoding CB₁R in the amygdala of adult marmosets through regional delivery of AAV-SaCas9-gRNA. We found that CB₁R knockdown in the amygdala induced anxiety-like behaviors, including disrupted night sleep, agitated psychomotor activity in new environments, and reduced social desire. Moreover, marmosets with CB₁R-knockdown had up-regulated plasma cortisol levels. These results indicate that the knockdown of CB₁Rs in the amygdala induces anxiety-like behaviors in marmosets, and this may be the mechanism underlying the regulation of anxiety by CB₁Rs in the amygdala of NHPs.
Animals ; Callithrix ; Receptors, Cannabinoid ; Anxiety ; Amygdala ; Cannabinoids ; Phenotype

OBJECTIVETo confirm the genetic diagnosis of two patients with ring chromosome 22 syndrome and investigate the mechanism underlying the formation of r(22) and potential genetic causes for the clinical phenotypes.

METHODSCytogenetic and molecular analyses using standard G-banding, fluorescence in situ hybridization and single nucleotide polymorphism array (SNP array) were performed.

RESULTSFor case 1, the karyotype was 46,XY,r(22)(p11q13). SNP array has identified a 7.0 Mb heterozygous deletion at 22q13.2q13.33. For case 2, the karyotype was 46,XY,r(22)(p11q13)[84]/45,XY,-22[6]; SNP array has detected a heterozygous microdeletion of 1.6 Mb at 22q13.33.

CONCLUSIONWith combined application of genetic testing, 2 cases of r(22) syndrome were diagnosed, which has improved the understanding of the genotype-phenotype correlation of r(22).

Child, Preschool ; Chromosome Banding ; Chromosomes, Human, Pair 22 ; genetics ; Genetic Testing ; Humans ; Male ; Nerve Tissue Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Ring Chromosomes

Objective Comparison of the levator ani muscles in three-dimensional (3D) MRI-based models in women with and without pelvic organ prolapse at rest to analyze the morphological characteristics of levator ani muscles in women with POP. Methods Twenty-five women with POP and 22 women with normal pelvic support were selected from Nanfang Hospital of Southern Medical University. Axial, sagittal, and coronal T2-weighted pelvic magnetic resonance scans were obtained with the women in the supine position.The 3D models were reconstructed from the source images. Morphological changes was compared within the two groups of levator ani muscles, and the 3D models were measured to determine the levator ani muscle volume (LVOL), levator plate angle (LPA), levator hiatus width (LH-W) and length (LH-L), distance between symphysis and levator sling muscle (LSG). Results There were no puborectalis avulsions in control, in POP, 3 cases of avulsions just in left, 3 cases of avulsions just in right, 7 cases in bilateral. The shape of iliococcygeus were all dome-shaped in control, 11 cases were U-shaped and 14 cases were dome-shaped in POP. The shape of levator hiatus were 7 cases of U-shape, 12 cases of V-shape, 3 cases of irregular in control; 5 cases of U-shape, 4 cases of V-shape, 16 cases of irregular in POP. POP versus control: LH-L: (68.0 ± 8.9) versus (61.6 ± 7.2) mm (P<0.05); LH-W: (41.4 ± 3.9) versus (38.0 ± 3.2) mm (P<0.05); LSG-L: (29.6 ± 7.4) versus (24.6 ± 3.7) mm (P<0.05); LSG-R: (28.4 ± 6.8) versus (23.9 ± 3.2) mm (P<0.05); LPA: (51.0 ± 11.3)° versus (40.6 ± 6.3)° (P<0.05); LVOL: (23.7 ± 5.8) versus (24.6 ± 5.0) cm3 (P>0.05). Conclusions It is possible to assess the morphologic changes of levator ani by using 3D MRI models objectively, our 3D data demonstrate larger in LVOL, LPA, LH-W, LH-L, LSG, and the changes in shape. It is helpful to diagnose and assess the specific situation of patients POP in clinic.

Objective To explore the application of 3D printing technology in individual pelvic structure of female and its value in obstetrics and gynecology based on the CT datasets of the pelvic structure and digital three-dimensional reconstruction. Methods CTA image dataset of a patient from gynecology department was obtained for constructing three-dimensional models of each organ using the digital three-dimensional reconstruction technology , then the digital 3D model with the same size as the model displayed was printed with Z510 3D printer. Results 3D models of patient′s lumbosacral vertebrae, aorta artery, common iliac artery, internal and external iliac artery , postcava , common iliac vein , internal and external iliac vein , pelvis ureter , uterus and uterine artery were printed out in the same size replica of the virtual reality model. Conclusion 3D printed model has all the features of 3D vision and can be touched and felt by people , which can provide new insights for medical education, clinical and medical research.