Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

Objective To explore the effect of repetitive transcranial magnetic stimulation(rTMS)on the sleep disorder and examination results of recruits.Methods At a training base,the Pittsburgh Sleep Quality Index(PSQI)was used to screen the recruits with sleep disorders(total score of PSQI>7).The recruits were randomly assigned to rTMS group or sham rTMS group.Both groups received cognitive and behavioral intervention therapy,including sleep health education and relaxation training.Moreover,the rTMS group was treated with rTMS at the right posterior parietal lobe by continuous theta burst stimulation(cTBS)twice a day at an interval of at least 50 min for 5 consecutive days as a course of treatment with an interval of 2 days for a total of 2 courses of treatment.The coil position and stimulus intensity of sham rTMS group were consistent with the rTMS group,but the head of subjects was perpendicular to the coil plane and there was no effective stimulation.Before and after treatment,PSQI,self-rating depression scale,generalized anxiety disorder-7 and health questionnaire-15 were used to evaluate the sleep,mood and physical state of the recruits.The training result was assessed one month after treatment.The total effective rate of PSQI improvement and examination results were compared between the two groups.The independent influencing factors of excellent examination result were analyzed.Results Among 351 recruits,83 with sleep disorders completed treatment and follow-up.There were 40 patients in the rTMS group and 43 patients in the sham rTMS group.There was no significant difference between the two groups at baseline.After treatment,the total effective rate of PSQI improvement in the rTMS group was higher than that in the sham rTMS group(77.50%vs 53.49%,P=0.022).The average examination score and excellent rate of the rTMS group were higher than those of the sham rTMS group(91.58±3.19 vs 89.47±4.67,P=0.020;85%vs 65.12%,P=0.037).Logistic regression analysis showed that the treatment mode(rTMS group)was the independent influencing factor of excellent examination results(P=0.032).Conclusion rTMS can effectively and safely improve the sleep disorders and examination results of recruits.rTMS may play a positive role in improving the learning and training effect of recruits,which needs to be further proved.

Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

BACKGROUND:Indolepropionic acid has been shown to reduce diabetes-induced central nervous system inflammation.However,there is a lack of research on whether to inhibit microglia M1 polarization for the treatment of spinal cord injury. OBJECTIVE:To investigate the mechanism of indolepropionic acid inhibition of microglial cell M1 polarization for the treatment of spinal cord injury through cell and animal experiments. METHODS:(1)In vitro experiments:BV2 cell viability was assessed using the CCK-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,BV2 cells were categorized into control group,administration group(50 μmol/L indolepropionic acid),lipopolysaccharide group(100 ng/mL lipopolysaccharide),and treatment group(100 ng/mL lipopolysaccharide + 50 μmol/L indolepropionic acid).Nitric oxide content was quantified using the Griess method.Real-time quantitative PCR and western blot assay were employed to measure mRNA and protein levels of pro-inflammatory factors.Cell immunofluorescence staining was conducted to assess inducible nitric oxide synthase expression.The Seahorse assay was employed to assess glycolytic stress levels in BV2 cells.(2)In vivo experiments:30 SD rats were randomly divided into three groups:sham surgery group,spinal cord injury group,and indolepropionic acid group.Motor function recovery in rats after spinal cord injury was assessed using BBB scoring and the inclined plane test.Immunofluorescence staining of spinal cord tissue was conducted to evaluate the expression of inducible nitric oxide synthase in microglial cells.ELISA was employed to measure protein expression levels of the pro-inflammatory cytokines interleukin-1β and tumor necrosis factor-α in spinal cord tissue. RESULTS AND CONCLUSION:(1)In vitro experiments:Indolepropionic acid exhibited significant suppression of BV2 cell viability when its concentration exceeded 50 μmol/L.Indolepropionic acid achieved this by inhibiting the activation of the nuclear factor κB signaling pathway,thereby suppressing the mRNA and protein expression levels of pro-inflammatory cytokines(interleukin-1β and tumor necrosis factor-α),as well as the M1 polarization marker,inducible nitric oxide synthase,in BV2 cells.Additionally,indolepropionic acid notably reduced the glycolytic level in BV2 cells induced by lipopolysaccharides.(2)In vivo experiments:Following indolepropionic acid intervention in spinal cord injury rats,there was a noticeable increase in BBB scores and the inclined plane test angle.There was also a significant decrease in the number of M1-polarized microglial cells in spinal cord tissue,accompanied by a marked reduction in the protein expression levels of pro-inflammatory cytokines(interleukin-1β and tumor necrosis factor-α).(3)These results conclude that indolepropionic acid promotes functional recovery after spinal cord injury by improving the inflammatory microenvironment through inhibition of microglia M1 polarization.

Objective:To investigate the associations of articular depression depth (ADD) and tibial plateau widening (TPW) by pre-operative CT measurement with incidence of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture.Methods:Included in this retrospective study were 131 patients who had been admitted to Emergency Center of Trauma, The Third Hospital Affiliated to Hebei Medical University from January 2016 to January 2020 for Schatzker type Ⅱtibial plateau fractures. They were 88 males and 51 females, aged from 18 to 60 years (average, 41.5 years), with 74 right and 57 left sides injured. All patients were treated with closed reduction and internal fixation assisted by bidirectional traction. Arthroscopy was used to detect the status of lateral meniscus immediately after closed reduction and internal fixation of the fracture fragments. Furthermore, patients were divided into 2 groups according to the integrity of lateral meniscus: meniscal tear group ( n=70) and tear-free group ( n=61). The 2 groups were compared in terms of age, gender, body mass index(BMI), injury side, time interval from injury to surgery, TPW and ADD. The receiver operating curve (ROC) was drafted to calculate the cut-off values of TPW and ADD in complication of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture. Results:The overall incidence of lateral meniscal tear in this cohort was 53.4% (70/131). There was no statistically significant difference in terms of age, gender, injury side, BMI or time interval from injury to surgery between the 2 groups ( P>0.05); TPW and ADD were significantly higher in the meniscal tear group than in the tear-free group ( P<0.05). To predict lateral meniscal tear in patients with Schatzker type Ⅱtibial plateau fracture, the area under ROC was 0.656 (95% CI: 0.562 to 0.750, P=0.002) for TPW and 0.709 (95% CI: 0.619 to 0.800, P<0.001) for ADD, respectively; the cut-off values of TPW and ADD were 4.3 mm and 6.1 mm. Conclusion:TPW and ADD may be effective predictors for prediction of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture.

Objective:To investigate the changes of white matter fiber bundles in patients with bipolar disorder depressive epoch.Methods:Forty-two patients with unmedicated bipolar disorder (BD) depression and 59 age-, sex- and handedness-matched healthy controls who underwent DTI were recruited in the study. According to the Johns Hopkins University human white matter fiber bundle map, the white matter tissue of the brain was segmented into 20 acknowledged large fiber bundles. The PANDA software was used to calculate the four average diffusion properties of each white matter fiber bundle for each subject. Nonparametric substitution test was used to detect the difference of diffusion index between the two groups on these 20 white matter fiber bundles. Pearson correlation analysis was performed between FA values and RD values extracted from significantly different white matter fiber bundles and clinical indices.Results:In comparison to the normal controls, BD patients had a significant decreased fractional anisotropy (FA) values in the left uncinate fasciculus (0.40±0.01 vs. 0.41±0.01, P=0.001) and the forceps minor (0.36±0.02 vs. 0.38±0.02, P<0.001). Additionally, the radial diffusivity values increased in the left uncinate fasciculus (6.57×10 -4±2.41×10 -5vs. 6.40×10 -4±2.42×10 -5, P=0.001 7). Pearson correlation analysis showed that there were no significant correlations among the clinical indices the FA values and AD values in the left uncinate fasciculus and forceps minor. Conclusions:The patients with bipolar disorder in depression possibly have abnormal left uncinate fasciculus and the forceps minor.

Objective:To investigate the changes of white matter fiber bundles in patients with bipolar disorder depressive epoch.Methods:Forty-two patients with unmedicated bipolar disorder (BD) depression and 59 age-, sex- and handedness-matched healthy controls who underwent DTI were recruited in the study. According to the Johns Hopkins University human white matter fiber bundle map, the white matter tissue of the brain was segmented into 20 acknowledged large fiber bundles. The PANDA software was used to calculate the four average diffusion properties of each white matter fiber bundle for each subject. Nonparametric substitution test was used to detect the difference of diffusion index between the two groups on these 20 white matter fiber bundles. Pearson correlation analysis was performed between FA values and RD values extracted from significantly different white matter fiber bundles and clinical indices.Results:In comparison to the normal controls, BD patients had a significant decreased fractional anisotropy (FA) values in the left uncinate fasciculus (0.40±0.01 vs. 0.41±0.01, P=0.001) and the forceps minor (0.36±0.02 vs. 0.38±0.02, P<0.001). Additionally, the radial diffusivity values increased in the left uncinate fasciculus (6.57×10 -4±2.41×10 -5vs. 6.40×10 -4±2.42×10 -5, P=0.001 7). Pearson correlation analysis showed that there were no significant correlations among the clinical indices the FA values and AD values in the left uncinate fasciculus and forceps minor. Conclusions:The patients with bipolar disorder in depression possibly have abnormal left uncinate fasciculus and the forceps minor.

Objective To explore the clinical features and risk factors of poor prognosis in neonatal necrotizing enterocolitis(NEC).Methods A retrospective study was carried out in the infants with NEC admitted to 6 cooperative hospitals in Guangdong Province between January 2005 and December 2014.The clinical features and risk factors of poor prognosis in preterm and full-term infants diagnosed NEC,early onset and late onset NEC were analyzed.Results A total of 449 cases who met the criteria were admitted during the study time.The mortality was 23.6％ (106/449 cases),of which the preterm group was 24.6％ (58/238 cases) while the full-term group was 22.7％ (48/211 cases),the early onset group was 22.1％ (45/204 cases) while the late onset group was 24.3％ (57/235 cases).The median number of NEC onset in preterm group was 11 d after birth while the number of the full-term group was 6 d.Full-term infants who diagnosed NEC were more likely to manifest themselves as abdominal distension (52.1％ vs.42.0％,x2 =4.597,P =0.032),vomiting(36.5％ vs.17.2％,x2 =21.428,P =0.000) and bloody stool(30.3％ vs.21.4％,x2 =4.653,P =0.031);but in the onset of NEC,preterm infants more likely to have feeding intolerance (21.0％ vs.12.8％,x2=5.309,P =0.021).The early onset group of full-term NEC was much common in twins or multiplets(9.4％ vs.1.1％,x2 =6.226,P =0.013),which rate of surgical therapy was much higher (41.0％ vs.27.0％,P =0.036) and the breast-feeding rate before NEC was lower than the late onset group(14.5％ vs.32.6％,x2 =9.500,P =0.002),the differences were statistically significant.The gestational age and birth weight were bigger in the early onset group of preterm NEC[(33.8 ±2.5) weeks vs.(32.2 ±2.8) weeks,t =4.261,P =0.000;(2.1 ±0.5) kg vs.(1.7 ± 0.5) kg,t =4.735,P =0.000)],but length of stay was shorter than the late onset group (18.0 d vs.26.5 d,P =0.000).Logistic regression analysis showed that the risk factors of poor prognosis of full-term NEC were shock,peritonitis and sepsis;while risk factors of poor prognosis of preterm NEC were small for gestational age infant,pulmonary hemorrhage,shock,intestinal perforation and sepsis;the risk factors of poor prognosis of the early onset group of full-term NEC was shock;while those of the late onset group were shock and peritonitis;the risk factors of poor prognosis in the early onset group of preterm NEC were shock and sepsis,while those in the late onset group were pulmonary hemorrhage,shock,intestinal perforation and sepsis.Conclusions Compared to the preterm NEC,the onset time of full-term NEC was earlier and the clinical manifestations were more typical.Early identification and management of shock,peritonitis,intestinal perforation,sepsis and pulmonary hemorrhage can reduce the risk of poor prognosis of neonate NEC.

A 50?year?old woman presented with intermittent dull pain in the forehead and mild dizziness occasionally after her forehead was subjected to a mild bump accidentally 20 days prior to the presentation, and was diagnosed with angioneurotic headache in a local hospital. After the treatment with oral sibelium tablets, the condition wasn′t relieved obviously. Computed tomography (CT) scan showed multiple localized bone destruction and low?density area in the frontal and bilateral parietal bones with adjacent soft tissue swelling. Magnetic resonance imaging(MRI)revealed equal T1 signals and slightly long T2 signals for multiple nodules in the frontal and bilateral parietal bones, high signals on diffusion?weighted imaging (DWI), obvious enhancement on contrast?enhanced MRI, and linear enhancement in adjacent meninges. Whole?body bone scintigraphy showed multiple increased radionuclide uptake in the skull. Laboratory examination demonstrated that specific antibodies to Treponema pallidum (Tp) were positive, and the serum rapid plasma reagin(RPR)titer was 1∶128. Cerebrospinal fluid(CSF)examination showed normal CSF pressure, nucleated cell counts(8 × 106/L)and glucose level(4.0 mmol/L), slightly high chloride flux(129.1 mmol/L), high protein level(0.9 g/L), high CSF?RPR titer of 1∶16 and presence of specific antibodies to Tp. Histopathological examination revealed hyperemia of adjacent tissues in the cranial osteolytic area, hyperplasia of interstitial fibrous tissue, endothelial cell swelling, and infiltration of inflammatory cells mainly containing plasma cells. The treatment regimen for neurosyphilis was given, and headache was relieved after 1 week of treatment, basically disappeared after 2 weeks, and completely disappeared after 4 weeks, and no similar headache occurred thereafter. Finally, the patient was diagnosed with acquired syphilitic skull osteomyelitis complicated by syphilitic meningitis.

Objective To prepare vardenafil hydrochloride orally disintegrating tablets and evaluate their quality. Methods The tablets were prepared by direct power compression method, using crosslinking povidone ( PVPP ) as disintegrants. The preparation method was optimized by response surface test using amount of PVPP, menthol and taste-masking agents as factors with disintegrating time and distance of bitterness as index. The results of taste of orally disintegrating tablets were determined by electronic tongue, comparing to the results of taste tests. At the same time, the properties of the tablets were evaluated using appearance, content uniformity, disintegrating time, et al. as index. Results The optimal formula was as follows:PVPP 13. 26%, menthol 0. 43%, taste-masking agent SGxj 1. 26%. The results on evaluation of electronic tongue were consistent with the results of taste tests. The quality of the prepared tablets was in line with standard. The disintegrating time was (22. 34 ± 0. 34 ) s. Conclusion The preparation technology of orally disintegrating tablets is simple, and controllable in quality.