[A quantitative MRI comparative study of imaging markers for cerebral small vessel disease in the middle-aged and elderly patients with and without hypertension].

Objective: To explore the difference of MRI markers of small cerebral vascular disease in middle-aged and elderly patients with hypertension and non-hypertension. Methods: A cross-sectional study. The clinical data of 316 patients who underwent head MRI with susceptibility weighted imaging scans at the Affiliated Zhongda Hospital of Southeast University from November 2013 to August 2022 were retrospectively analyzed, including 190 males and 126 females, with the age of (71.6±8.9)years. According to the history of hypertension, the patients were divided into hypertension group(n=259) and the non-hypertension group(n=57). The patients in the non-hypertension group were further divided into abnormal blood pressure group on admission (n=19) and normal blood pressure group on admission (n=38). The imaging features of different CSVD dimensions in the patient's images were quantified or graded and compared between hypertensive and non-hypertensive patient groups. Deep learning methods were employed to segment white matter lesions, and voxel-wise analysis was conducted to investigate the differences in whole-brain white matter lesion probability between patients in both groups. Spearman correlation analysis was used to analyze the correlation between hypertension and small cerebral vascular disease. Results: Compared with the non-hypertensive group, the cerebral microhemorrhage count, deep microhemorrhage count, basal ganglia level lacunae count, perivascular space (EPVS) grade of hemioval center level and EPVS grade of basal ganglia level were higher in the hypertensive group (all P<0.05). The cerebral microhemorrhage count [3.0(1.0, 15.0) vs 1.0 (0, 4.2)], deep microhemorrhage count [1.0 (0, 7.0) vs 1.0 (0, 4.2)] and EPVS classification at basal ganglium level [2.0(1.0, 3.0) vs 1.0(1.0, 2.0)] in the group with history of hypertension were higher than those in the group with normal blood pressure at admission (all P<0.05). The EPVS grade at the central level of the semiovale in the hypertension group was higher than that of the group with normal blood pressure at admission [2.0(1.0, 2.0) vs 1.0 (1.0, 2.0)], and also higher than that of the group with abnormal blood pressure at admission [2.0(1.0, 2.0) vs 1.0(1.0, 2.0)](both P<0.05). Voxel-by-voxel analysis showed no significant difference in the probability of white matter lesions in the whole brain between patients with and without a history of hypertension, but patients with a history of hypertension showed more extensive para-ventricular white matter hypersignaling than those without a history of hypertension. Spearman correlation analysis showed that hypertension grade was correlated with the number of microbleeding lesions in depth (r=0.149), the number of lacunae lesions in the center of the hemioval (r=0.209), and the number of lacunae lesions in the basal ganglia (r=0.204) (all P<0.05). Conclusions: Chronic hypertension can affect different dimensions of small vessel disease imaging, primarily manifested in the increases of deep microbleed counts and the EPVS grade.

[Analysis of the effect of percutaneous vertebroplasty combined with (125)I seed implantation in the treatment of spinal metastatic epidural spinal cord compression].

Objective: To investigate the clinical efficacy of percutaneous vertebroplasty (PVP) combined with iodine-125 ((125)I) seed brachytherapy in the treatment of spinal metastatic epidural spinal cord compression (MESCC) and toassess the changes inthe grade of epidural spinal cord compression (ESCC) by magnetic resonance imaging (MRI). Methods: A total of 37 MESCC patients treated with PVP combined with (125)I seed brachytherapy in the interventional and vascular surgery department of Zhongda Hospital affiliated to Southeast University from January 2014 to June 2019 were retrospectively analyzed, including 23 cases of bilateral lower limbs paralysis. Total diseased vertebrae are 39 segments. Visual analogue scale (VAS) and paralysis of lower extremities were evaluated regularly before and after treatment, and VAS values at different follow-up time points were compared. At the same time, MRI was used to evaluate the changes of ESCC grade in the spinal canal and calculate the local lesion efficiency after operation. The postoperative local lesion efficiency at different follow-up times was compared. Results: PVP combined with (125)I seed implantation in all diseased vertebral bodies was successful. The average injection volume of polymethylmethacrylate (PMMA) was (3.2±1.3) ml/segment, the average number of (125)I seed implanted was (25.0±8.6) seeds/segment and the average radiation dose was (15.0±5.1) mCi/segment. The VAS before operation was 8.5, and postoperative VAS were respectively 3.6±1.3, 3.8±1.5, 3.4±1.4, 5.5±1.0, 5.9±1.4 at 5 days, 1 month, 3 months, 6 months, and 1 year after operation. The differences between all follow-up time points and preoperative VAS values were statistically significant (all P<0.001). Compared with 5 days, 1 month and 3 months after operation, VAS increased significantly at 6 months and 1 year after operation, and the difference was statistically significant (all P<0.001); there was no significant difference between the VAS value at 6 months after operation and 1 year after operation (P=0.405). At a follow-up of 3 months, 22 of 23 patients with paralysis of bilateral lower limbs regained the functions of autonomous walking and voiding; the effective rates of MESCC local lesions evaluated by MRI at 1 month, 3 months, 6 months, and>1 year were 89.7%, 91.9%, 90.6%, and 94.7%, respectively, and there was no statistically significant differences among those follow-up time points (all P>0.05). Conclusions: PVP combined with (125)I seed brachytherapy in the treatment of MESCC has significant improvement in immediate pain relief and spinal cord function. After combined treatment, MRI showed that the tumors around the spinal cord regressed dramatically, which could considerably reduce the MESCC grade and remain stable for a long time.

[Occlusal changes before and after orthodontic treatment in patients with aggressive periodontitis].

OBJECTIVE: To evaluate the efficacy of occlusal improvement in the labial fixed orthodontic treatment in aggressive periodontitis patients and to explore the relationship between occlusal improvement and inflammation control. METHODS: Twenty-two aggressive periodontitis patients who underwent combined periodontal-orthodontic treatment were included in this study. The patient's photos were matched to the dental models and digital three dimentional models were acquired using 3Shape R700 laser scanner. The occlusal force distribution maps were generated in the OrthoAnalyzer software. The newly established occlusal force distribution score (OFDS) and proximal contact score (PCS) were used to evaluate the occlusal distribution changes before and after labial fixed orthodontic treatment for assessing the effectiveness of orthodontic treatment. The multi-level linear regression analysis was used to explore the relationship between the probing depth changes and OFDS or PCS changes to screen out the favorable orthodontic strategy for inflammation control, which would provide clinical strategy for combined periodontal-orthodontic treatment in aggressive periodontitis patients. RESULTS: At the patient level, OFDS was improved significantly after orthodontic treatment compared with the score before orthodontic treatment (84.5±20.9 vs.105.3±22.6, P <0.001) and PCS was improved significantly after orthodontic treatment compared with the score before orthodontic treatment (68.9±9.1 vs. 83.7±6.3, P <0.001).At the tooth level, the OFDS was significantly increased in the maxillary anterior teeth (P <0.001) while the PCS of the anterior teeth in both maxillary and mandible arches were significantly increased significantly (P <0.01). No significant changes were found in other tooth positions. The multilevel linear regression model showed that no significant correlation was found between age and gender and probing depth decrease (P >0.05). The baseline probing depth,OFDS improvements and PCS improvements (P <0.001) were positively correlated with probing depth decrease. CONCLUSION: This study showed that the distribution of occlusal force was more reasonable and the proximal contacts were more ideal in aggressive periodontitis patients. Orthodontic treatment was effective in improving occlusal force distribution by the above two ways. Especially, the OFDS and PCS improvements were both positively correlated with probing depth decrease, indicating that in the combined periodontal-orthodontic treatment for aggressive periodontitis patients, occlusal force distribution and proximal contact should be improved in order to facilitate periodontal improvement.

[Cement augmentation for vertebral osteolytic metastatic lesions: an evaluation on postoperative CT].

Objective: To evaluate the effectiveness of cement augmentation on the osteolytic lesion in patients with vertebral metastasis. Methods: A total of consecutive 132 patients with 268 vertebral metastatic lesions treated with PVP from January 2008 to December 2016 in Zhongda Hospital were enrolled in this study. Retrospective analysis of preoperative, postoperative 3 days, 3 months, 6 months, 12 months and ≥ 18 months imaging data on CT, the local control and progression of the tumor were evaluated by MDA response criteria. The local control rates were compared between the groups with the different rate of cement filling by Chi2-test. Results: Vertebroplasty procedures were performed successfully in all 268 vertebrae under DSA guidance, and the mean volume of PMMA injected in each vertebra was 0.7-8.5(3.9±1.5)ml.The rate of local control at 3 months, 6 months, 12 months and ≥18 months after PVP was respectively 98.9%, 95.1%, 91.8%, and 85.2%, the difference was statistically significant(all P<0.05). The local control rate showed a statistically significant relationship to the groups with the rate of cement filling at 6 months, 12 months and ≥18 months after PVP, but there was no statistical difference at postoperative 3 months. The rate of local control was higher in 68 patients with lung or gastrointestinal cancer than in 17 patients with liver or kidney cancer at 3 months, 6 months and 12 months, the difference was statistically significant (P<0.05). Conclusion: Cement augmentation has a local anti-tumor effect on vertebral osteolytic metastatic lesion, and the anti-tumor effect will decrease as the follow-up time extended.

[Oblique puncture approach via anterolateral to C2: a study of imaging anatomy on corpses].

Objective: To determine the anatomical relation between the carotid artery or laryngopharynx and the needle path via anterolateral oblique approach to C2 vertebral body, and whether the simulated path can direct the actual procedure. Methods: Contrast agent was injected into bilateral carotid arteries of 10 fresh cadavers (20 sides), and then CT scans were performed on each subject. Bilateral simulated puncture paths were simulated on the image of two-dimensional reconstruction CT. The distance between the puncture site and middle line(AC), the angle between the simulated puncture paths and the median sagittal plane (AOB), the distance between the simulated puncture paths and the side wall of hypopharynx, or the carotid artery was measured respectively. Under the guidance of fluoroscopy, oblique puncture via anterolateral to C2 was performed according to the simulated parameters, and then CT was done. The actual needle path was measured on the image of two-dimensional reconstruction CT. The outcomes were compared by using paired t test between groups. Results: Bilateral simulated puncture paths in all 10 cadavers were achieved on the image of CT. The bilateral punctures via anterolateral to C2 were successfully performed under fluoroscopic guidance in 10 cadavers, and the actual needle path could be showed on the image of two-dimensional reconstruction CT. The AC, angle AOB, the distance to carotid artery and side wall of laryngopharyngeal of the left simulated path was(4.7±1.0)cm, (33±8)°, (6.7±2.6)mm, (6.6±2.8)mm, respectively; and those in the right side was(5.2±1.3) cm, (36±8)°, (7.5±2.2) mm, (7.2±2.5) mm, respectively.The AC, angle AOB, the distance to carotid artery and side wall of laryngopharyngeal of the left actual needle was(4.5±1.1) cm, (33±9) °, (6.7±3.0) mm, (7.1±3.9) mm, respectively; and those in the right side was(5.1±1.5) cm, (35±10) °, (7.4±2.1) mm, (7.2±2.6) mm, respectively.And the actual and simulation data of both sides were compared and no significant differences was found by t test (t=0.104-1.882, all P>0.05). Conclusion: The simulated approach via anterolateral to C2 can be achieved on the image of two-dimensional reconstruction CT, it is consistent with the real needle path and can be used to direct the actual practice to improve the accuracy and safety.

[Expression of hepatitis E virus ORF3 gene fragment in baculovirus system and its immunological character].

OBJECTIVE: To express hepatitis E virus (HEV) ORF3 protein by baculovirus system and provide basis for immunological character research. METHODS: Hepatitis E Virus ORF3 gene fragment was obtained by RT-PCR, ligated with vector pThioHisA for sequencing and then inserted into transfected vector pVL1393 to construct recombinant plasmid. Mediated by Lipofectin Reagent, the recombinant vector and baculovirus linearized DNA (BaculoGold) co-transfected insect cell Sf9 to make recombinant baculovirus. Expressed ORF3 was analyzed for its immunological character by Western blotting, and immunized Kunming Mice. RESULTS: Recombinant ORF3 protein could be recognized by the known positive serum and promoted organism to produce HEV-specific antibody. CONCLUSIONS: Recombinant baculovirus can express effectively HEV ORF3, which has HEV specific immunogenic character.

[Purification and immunological characterization of hepatitis E virus recombinant chimeric antigen encoded by ORF2 fragments and ORF3].

OBJECTIVE: To study immunological characteristics of recombinant chimeric HEV antigen. METHODS: Constructed recombinant plasmids pThioHisORF(2.1 + 2.2 + 3), which contains three HEV antigen gene fragments (ORF2.1:6287-6403nt, ORF2.2:6743-7126nt, ORF3), was transformed into E. coli and induced with IPTG. Expressed product P(2.1 + 2.2 + 3) existed in inclusion bodies, was purified by denature SP Sepharose FF cation exchange chromatography. Rabbits and rats were immunized with renatured P(2.1 + 2.2 + 3). The level of IgG in sera from experimental animals and clinical patients were examined with P(2.1 + 2.2 + 3) by ELISA. The characteristics of IgG of immunized animals interacted with recombinant antigen expressed by baculovirus system as well as recombinant chimeric antigen interacted with clinical patients sera were evaluated by Western-blotting. RESULTS: High titer of IgG antibodies, 1:25,600 in rabbits and 1:12,800 in rats, were detected after immunized with P(2.1 + 2.2 + 3). Furthermore, recombinant antigen expressed by baculovirus system was specifically recognized by IgG of experimental animal immunized with P(2.1 + 2.2 + 3), and the purified recombinant chimeric antigen P(2.1 + 2.2 + 3) was specifically reacted with the IgG of clinical patients. CONCLUSIONS: Recombinant chimeric antigen appears a promising strategy for detection of and prevention from HEV infection.

[Propagation of hepatitis E virus in several cell lines including human embryo lung diploid cell KMB17].

OBJECTIVE: To investigate the hepatitis E virus (HEV) sensitive cells and its tissue culture conditions. METHODS: The HEV from dejecta supernatant of patients with acute hepatitis E was amplified and activated by passaged in Rhesus. Then, the positive dejecta samples of infected monkeys were dealt with super-centrifugation and virus for culture was obtained. Various human-derived (including KMB17, A549, BEL7402, and Hela) and non-human primates derived cells (Vero) were inoculated with HEV. Sensitivity of cells to HEV was measured by CPE (cytopathic effect), RT-PCR and immunofluorescence. RESULTS: CPE in KMB17, A549 and BEL7402 cells appeared during 7-9 days, meanwhile, cells shelled during 11-13 days on the first filial generation. The existence of HEV genome +RNA and replicated -RNA was still detectable by RT-PCR after the tenth filial generation. Neither CPE nor amplification of HEV genome RNA could be detected in Hela and Vero cells after the second to fourth filial generation. HEV could also be detected from inoculated KMB17 cells by immunofluorescence and RT-PCR. CONCLUSIONS: It indicates that KMB17, A549 and BEL7402 cells are sensitive to HEV under the experimental culture conditions, while Hela and Vero cells are insensitive. Tissue culture system of HEV in certain filial generation is established.

Deterioration of mechanical properties of discs in chronically inflamed TMJ.

Temporomandibular joint (TMJ) discs frequently undergo degenerative changes in arthritis. However, the biomechanical properties of pathogenic discs remain to be explored. In this study, we evaluated the effects of chronic inflammation on the biomechanical properties of TMJ discs in rats. Chronic inflammation of TMJs was induced by double intra-articular injections of complete Freund's adjuvant for 5 weeks, and biomechanical properties and ultrastructure of the discs were examined by mechanical testing, scanning electron microscopy, and transmission electron microscopy. The instantaneous compressive moduli of the anterior and posterior bands of discs in inflamed TMJs were decreased significantly compared with those in the control group. The instantaneous tensile moduli of the discs of inflamed TMJs also showed significant decreases in both the anterior-posterior and mesial-lateral directions. The relaxation moduli of the discs of inflamed TMJs showed nearly the same tendency as the instantaneous moduli. The surfaces of the discs of inflamed TMJs became rough and porous due to the loss of the superficial gel-like stratum, with many collagen fibers exposed and degradation of the sub-superficial collagen fibrils. Our results suggested that chronic inflammation of TMJ could lead to deterioration of mechanical properties and alteration of disc ultrastructure, which might contribute to TMJ disc displacement.

A novel serine protease inhibitor acts as an immunomodulatory switch while maintaining homeostasis.

Serine protease cascades boost immune responses while maintaining homeostasis. These crucial actions are intricately regulated by cognate serine protease inhibitors. However, the mechanism underlying such a dynamic immunomodulation during acute phase infection remains obscure, particularly where the pathogen's serine protease adds a new challenge to the host. Here, we found that infection of horseshoe crab, Carcinoscorpius rotundicauda, induced reciprocal profiles of CrSPI (serine protease inhibitor) and CrFurin (serine protease) with respect to their transcription and protein activities. Using recombinant rCrSPI, we explored its inhibitory activity against various microbial proteases and found it most efficacious against a model serine protease, subtilisin A. rCrSPI inhibited subtilisin at Ki 10(-9)M with a molar ratio of 1 rCrSPI:2 subtilisin. The rCrSPI also inhibited plasma CrFurin, suppressed subtilisin-mediated activation of prophenoloxidase (PPO) and interacted with complement C3. Taken together, CrSPI acts as a key immunomodulatory 'on-off' switch in a 2-way regulation of serine protease microbial subtilisin and host serine proteases (CrFurin and CrC3), thereby controlling immune responses involving the complements and the PPO-mediated antimicrobial activities, while maintaining homeostasis.