The DNA fragments of ag85b、esat-6、hsp65、mpb64 and ag85b-esat-6、hsp65-esat-6、mpb64-esat-6 were amplified by PCR and SOE technique.These seven fragments were inserted into pCDNA3.1(+)vector to construct recombinant plasmids pCA、pCE6、pCH、pCM、pCAE、pCHE and pCME.The seven plasmids were transfected into SP2/0 cell in vitro to detect the expression of target genes.BALB/c mice were intramuscularly vaccinated with the seven plasmids and the control vector pCDNA3.1(+)and PBS respectively.The serum antibodies and the spleen lymphocyte proliferation(SLP)and secreted IFN～? of spleen were tested.The results of indirect ELISA showed the levels of antibodies in all recombinant plasmids groups were significantly higher than the two control groups(P

Background Proline hydroxylase 2 (PHD2) is a degrading enzyme of hypoxia-inducible factor-1α(HIF-1α) and can regulate the expression of vascular endothelial growth factor (VEGF) and erythropoietin that promote hypoxia-induced blood vessel generation.Baicalein is an inhibitor of PHD2.Understanding the inhibitory effects of baicalein on overexpression of PHD2 in hypoxia-induced retinal glial cells is helpful for elucidating the pathogenesis of the ocular nevascular diseases.Objective This study aimed to observe the expression change of PHD2 in hypoxia-induced retinal glial cells and explore the inhibition of baicalein on PHD2.Methods The study protocol was approved by Experimental Animal Ethic Committee.Retinal glial cells were isolated from SPF neonatal SD rats (within 48 hours) and primarily cultured in DEME by explant cell culture.Cultured cells were identified by immunofluorescence staining of glial fibrillary acidic protein.The cells were divided into normal control group,CoCl2 group and CoCl2 +baicalein group.The cells in the normal control group were cultured in the conventional medium,and 200 μmol/L CoCl2 was added in the medium to establish the hypoxia cell models in the CoCl2 group,and 200 μmol/L CoCl2 and 50 μmol/L baicalein were added in the medium in the CoCl2 + baicalein group.The proliferation (absorbancy) of the cells was detected by cell counting kit-8 (CCK-8).The expression of PHD2 and VEGF in the cells was detected and located using immunofluorescence staining.The expressions of PHD2 mRNA and VEGF mRNA and their proteins in the cells were assayed by real-time quantitative PCR and Western blot,respectively.Resnlts The proliferation (absorbancy) of the cells was significantly different among the groups (F=132.05,P=0.00),and the absorbancy was considerably increased in the CoCl2 group compared with normal control group and CoCl2 + baicalein group,with significant differences between the groups (both at P＜ 0.01).Immunofluorescence staining showed that the PHD2 and VEGF were weakly expressed in the cells of the normal control group,and the expression intensity was high in the CoCl2 group.The expression intensities of PHD2 and VEGF in the cells were weakened in the CoCl2 +baicalein group compared with those of the CoCl2 group.The relative expression levels of PHD2 mRNA were 0.366±0.034,0.894 ± 0.015 and 0.445 ± 0.017 and those of PHD2 protein were 131.27 ± 2.61,140.12 ± 4.29,133.14±2.11;those of VEGF mRNA were 1.346 ± 0.008,3.465 ± 0.048 and 2.264 ± 0.073 and those of VEGF protein were 532.25 ± 19.92,601.13 ± 10.21,537.34 ± 5.96 in the normal control group,CoCl2 group and CoCl2 + baicalein group,showing significant differences among the three groups (PHD2:F =905.89,43.18,both at P＜0.01.VEGF:F=27.13,185.79,both at P ＜ 0.01),and the relative expression levels of PHD2 and VEGF mRNA and protein in the CoCl2 group were higher than those in the normal control group and CoCl2 +baicalein group (all at P＜ 0.01).Conclusions Hypoxia stimulates the proliferation of retinal glial cells and increase the expression of PHD2 and VEGF.Baicalein can effectively inhibit the overexpression of PHD2 and VEGF in retinal glial cells and proliferation of retinal glial cells induced by hypoxia.PHD2 is a potential provention and treatment target of hypoxiainduced retinal neovascularization.

OBJECTIVE: To propose a sensitivity test method for geometric correction position deviation of cone-beam CT systems. METHODS: We proposed the definition of center deviation and its derivation. We analyzed the influence of the variation of the three-dimensional spatial center of the steel ball point, the projection center and the size of the steel ball point on the deviation of geometric parameters and the reconstructed image results by calculating the geometric correction parameters based on the Noo analytical method using the FDK reconstruction algorithm for image reconstruction. RESULTS: The radius of the steel ball point was within 3 mm. The deviation of the center of the calibration parameter was within the order of magnitude and negligible. A 10% Gaussian perturbation of a single pixel in the 3D spatial coordinates of the steel ball point produced a deviation of about 3 pixel sizes, while the same Gaussian perturbation of the 2D projection coordinates of the steel ball point produced a deviation of about 2 pixel sizes. CONCLUSION The geometric correction is more sensitive to the deviation generated by the three-dimensional spatial coordinates of the steel ball point with limited sensitivity to the deviation generated by the two-dimensional projection coordinates of the steel ball point. The deviation sensitivity of a small diameter steel ball point can be ignored.
Algorithms ; Calibration ; Cone-Beam Computed Tomography ; Steel

BACKGROUND: Textbook outcome (TO) can guide decision-making among patients and clinicians during preoperative patient selection and postoperative quality improvement. We explored the factors associated with achieving a TO for gallbladder carcinoma (GBC) after curative-intent resection and analyzed the effect of adjuvant chemotherapy (ACT) on TO and non-TO patients. METHODS: A total of 540 patients who underwent curative-intent resection for GBC at the Department of Hepatobiliary Surgery of the First Affiliated Hospital of Xi'an Jiaotong University from January 2011 to December 2020 were retrospectively analyzed. Multivariable logistic regression was used to investigate the factors associated with TO. RESULTS: Among 540 patients with GBC who underwent curative-intent resection, 223 patients (41.3%) achieved a TO. The incidence of TO ranged from 19.0% to 51.0% across the study period, with a slightly increasing trend over the study period. The multivariate analysis showed that non-TO was an independent risk factor for prognosis among GBC patients after resection ( P = 0.003). Age ≤60 years ( P = 0.016), total bilirubin (TBIL) level ≤34.1 μmol/L ( P <0.001), well-differentiated tumor ( P = 0.008), no liver involvement ( P <0.001), and T1-2 stage disease ( P = 0.006) were independently associated with achieving a TO for GBC after resection. Before and after propensity score matching (PSM), the overall survival outcomes of non-TO GBC patients who received ACT and those who did not were statistically significant; ACT improved the prognosis of patients in the non-TO group ( P <0.05). CONCLUSION Achieving a TO is associated with a better long-term prognosis among GBC patients after curative-intent resection, and ACT can improve the prognosis of those with non-TO.
Humans ; Middle Aged ; Gallbladder Neoplasms/pathology* ; Retrospective Studies ; Prognosis ; Hepatectomy ; Cholecystectomy

ObjectiveTo observe the effect of enriched environment (EE) combined with acupuncture at head point (HA) on behavior in rats with autism spectrum disorder. MethodsHealthy female Wistar rats were given peritoneal injection of sodium valproate at 12.5 days of gestation. Twenty-four male offspring rats were randomly selected and then randomly divided into model group (n = 6), EE group (n = 6), HA group (n = 6) and EE combined with HA group (the combined group, n = 6). Six male offspring rats born from female mice injected with the same amount of saline intraperitoneally were as control group. After four weeks of treatment, all the five groups were tested with three-chamber test and marble burying test, and the sociability index, the social novelty index and the number of buried marbles were recorded. The levels of interleukin (IL)-1β and IL-6 in peripheral blood were determined by enzyme-linked immunosorbent assay (ELISA). ResultsAfter treatment, compared with the model group, the sociability index and the social novelty index improved (P < 0.05), the number of buried marbles reduced (P < 0.05), and the levels of IL-6 and IL-1β in peripheral blood decreased in EE group, HA group and the combined group (P < 0.05); while the combined group was the best (P < 0.01). ConclusionBoth EE or acupuncture at HA could improve behavioral symptoms, and reduce the expression of inflammatory factors in rats with autism spectrum disorder. The combination of the two methods showed the best result.