Protective Mechanism of Paeoniflorin on Mice with Ulcerative Colitis Based on AMPK/mTOR Autophagy Pathway / 中国实验方剂学杂志

ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis （UC） through the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS， and 56 BALB/c male mice were randomly divided into model group， AMPK inhibitor group （20 mg·kg-1）， paeoniflorin （50 mg·kg-1） + inhibitor （20 mg·kg-1） group， and high dose （50 mg·kg-1）， medium dose （25 mg·kg-1）， and low dose （12.5 mg·kg-1） paeoniflorin groups. After seven days of drug intervention， the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight， disease activity index （DAI） changes， and Hematoxylin-eosin （HE） staining results. Enzyme linked immunosorbent assay （ELISA） was used to detect the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the serum of mice in each group， and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 （LC3） content in the colon， AMPK， mTOR proteins， and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot， and the mRNA expression levels of AMPK， mTOR， Beclin1， LC3， and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the model group showed a decrease in body mass， an increase in DAI score， and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum （P<0.01）， while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue， and those of p-mTOR/mTOR were up-regulated （P<0.01）. The mRNA expression levels of AMPK and LC3 were down-regulated， while the mRNA expression levels of mTOR and p62 were up-regulated （P<0.01）. Compared with the model group and the paeoniflorin + inhibitor group， the mice treated with paeoniflorin showed an increase in body mass， a decrease in DAI score， a reduction in pathological damage to colon tissue， and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum （P<0.05）. The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated， while the protein levels of p-mTOR/mTOR were down-regulated （P<0.01）. The mRNA expression levels of AMPK， Beclin1， and LC3 were up-regulated， while the mRNA expression of mTOR and p62 were down-regulated （P<0.01）. The colon tissue of the inhibitor group was severely damaged， and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.

Development and validation of a CT-based radiomics model for differentiating pneumonia-like primary pulmonary lymphoma from infectious pneumonia: A multicenter study / 中华医学杂志(英文版)

BACKGROUND@#Pneumonia-like primary pulmonary lymphoma (PPL) was commonly misdiagnosed as infectious pneumonia, leading to delayed treatment. The purpose of this study was to establish a computed tomography (CT)-based radiomics model to differentiate pneumonia-like PPL from infectious pneumonia.@*METHODS@#In this retrospective study, 79 patients with pneumonia-like PPL and 176 patients with infectious pneumonia from 12 medical centers were enrolled. Patients from center 1 to center 7 were assigned to the training or validation cohort, and the remaining patients from other centers were used as the external test cohort. Radiomics features were extracted from CT images. A three-step procedure was applied for radiomics feature selection and radiomics signature building, including the inter- and intra-class correlation coefficients (ICCs), a one-way analysis of variance (ANOVA), and least absolute shrinkage and selection operator (LASSO). Univariate and multivariate analyses were used to identify the significant clinicoradiological variables and construct a clinical factor model. Two radiologists reviewed the CT images for the external test set. Performance of the radiomics model, clinical factor model, and each radiologist were assessed by receiver operating characteristic, and area under the curve (AUC) was compared.@*RESULTS@#A total of 144 patients (44 with pneumonia-like PPL and 100 infectious pneumonia) were in the training cohort, 38 patients (12 with pneumonia-like PPL and 26 infectious pneumonia) were in the validation cohort, and 73 patients (23 with pneumonia-like PPL and 50 infectious pneumonia) were in the external test cohort. Twenty-three radiomics features were selected to build the radiomics model, which yielded AUCs of 0.95 (95% confidence interval [CI]: 0.94-0.99), 0.93 (95% CI: 0.85-0.98), and 0.94 (95% CI: 0.87-0.99) in the training, validation, and external test cohort, respectively. The AUCs for the two readers and clinical factor model were 0.74 (95% CI: 0.63-0.83), 0.72 (95% CI: 0.62-0.82), and 0.73 (95% CI: 0.62-0.84) in the external test cohort, respectively. The radiomics model outperformed both the readers' interpretation and clinical factor model ( P <0.05).@*CONCLUSIONS@#The CT-based radiomics model may provide an effective and non-invasive tool to differentiate pneumonia-like PPL from infectious pneumonia, which might provide assistance for clinicians in tailoring precise therapy.

Imaging features and clinical outcome in fetus with coronal cleft vertebrae / 中华放射学杂志

Objective:To explore the MRI features of fetal coronal cleft vertebrae, and to compare the efficacy of MRI and ultrasound in the diagnosis of fetal coronal cleft vertebrae, and to analyze the outcome of fetal coronal cleft vertebrae.Methods:From September 2019 to June 2021, 40 fetuses suspected of fetal vertebral deformities by ultrasound were retrospectively collected in Shandong Provincial Hospital Affiliated to Shandong University, who were diagnosed as coronal cleft vertebrae after MRI examination. Five cases of induced labor and 14 cases lost to follow-up were excluded, and 21 fetuses who underwent MRI after delivery were finally included. The gestational weeks were 25-34 (29.1±2.6) weeks, and there were 19 males and 2 females. Fetal spine MRI includes susceptibility weighted imaging (SWI) and T 2-true fast imaging with steady-state (True-FISP). The MRI features and outcome of fetal coronal cleft vertebrae were explored. The image quality scores of SWI, T 2-True-FISP and ultrasound were compared with Friedman test and Wilcoxon signed-rank test. The diagnostic accuracy of fetal coronal cleft vertebrae of SWI, T 2-True-FISP and ultrasound was calculated. Cochran test was used to compare the efficiency of 3 kinds of images, and the modified McNemar test was used for pairwise comparison between groups. Results:There were 10 cases of single and 11 cases of multiple fetal coronal cleft vertebrae, 16 cases of simple lumbar vertebrae, 2 cases of simple thoracic vertebrae, and 3 cases of thoracolumbar vertebrae. The common SWI features of 21 cases show longitudinal strip or dot high signal on the sagittal plane, and transverse fissure like high signal on axial plane. Anterior part of vertebral body was larger than posterior part in 19 cases of them. The image quality scores of SWI, T 2-True-FISP and ultrasound were 4 (3, 4), 2 (2, 2), 2 (2, 2), and the difference was statistically significant in general (χ2=34.24, P<0.001). Pairwise comparison showed that the image quality of SWI was better than those of T 2-True-FISP and ultrasound ( Z=-4.04, P<0.001; Z=-4.11, P<0.001), and there was no statistically significant difference between T 2-True-FISP and ultrasound ( Z=-0.58, P=0.388). The diagnostic accuracy of SWI, T 2-True-FISP and ultrasound was 100% (21/21), 66.7% (14/21), 47.6% (10/21). The diagnostic accuracy of SWI was better than those of T 2-True-FISP and ultrasound (χ2=5.14, P=0.008; χ2=9.09, P<0.001), and there was no statistically significant difference between T 2-True-FISP and ultrasound (χ2=0.75, P=0.194). MRI showed that coronal cleft vertebrae disappeared in all 21 fetuses after birth, including 1 case of syringomyelia and 1 case of fatty filum terminal. Conclusions:MRI, especially SWI, plays an important role in the diagnosis of fetal coronal cleft vertebrae. Fetal coronal cleft vertebrae disappeared in the follow-up after birth, which proved to be a normal physiological variation from the radiographic perspective.

Effect of Shenmai Injection on Long-Term Prognosis of Patients with Chronic Heart Failure: A Multicenter, Large Sample Capacity, Long-Term Follow-Up Retrospective Cohort Study / 中国结合医学杂志

OBJECTIVE@#To explore the effect of Shenmai Injection (SMI) on the long-term prognosis of patients with chronic heart failure (CHF).@*METHODS@#The Hospital Information System was used to extract data of CHF patients, and the retrospective cohort study was conducted for analysis. In non-exposed group, standardized Western medicine treatment and Chinese patent medicine or decoction were applied without combination of SMI while in the exposed group, SMI were applied for more than 7 days. Evaluation indicators are followed with New York Heart Association functional classification (NYHA classification), left ventricular ejection fraction (LVEF), N-terminal brain natriuretic peptide precursor (NT-ProBNP), cardiogenic death and heart failure (HF) readmission. Statistical analysis includes Kaplan-Meier analysis and Cox regression which are used to explore the relationship between SMI and outcome events.@*RESULTS@#A total of 1,211 eligible CHF patients were involved and finally 1,047 patients were followed up successfully. After treatment, the cases of NYHA classification decline in the exposed and non-exposed groups accounted for 64.30% and 43.45%, respectively; the improvement values of LVEF were 8.89% and 7.91%, respectively; the improvement values of NT-ProBNP were 909 pg/mL and 735 pg/mL, respectively. After exposure on SMI, the rates of cardiogenic death and HF readmission reduced from 15.43% to 10.18% and 38.93% to 32.37%. According to Kaplan-Meier analysis, the log-rank P value of SMI and cardiogenic death was 0.014, while the counterpart of SMI and HF readmission was 0.025. Cox regression analysis indicated that for cardiogenic death, age, cardiomyopathy, diabetes, and NYHA classification were risk factors while β-blockers, aldosterone receptor antagonists, Chinese patent medicine/decoction and SMI were protective factors. Likewise, for HF readmission, age, cardiomyopathy, and NYHA classification were risk factors while SMI was a protective factor.@*CONCLUSION@#Combination with SMI on the standardized Western medicine treatment can effectively reduce cardiogenic mortality and readmission rate in CHF patients, and thereby improve the long-term prognosis.

The value of MRI anterior cartilaginous acetabulum-head-index to evaluate hip function after treatment of developmental dysplasia of the hip / 中华放射学杂志

Objective:To evaluate the feasibility of high resolution MRI for the measurement of anterior cartilaginous acetabulum-head-index (A-CAHI) and the value of A-CAHI for predicting hip clinical function after treatment in developmental dysplasia of the hip (DDH).Methods:The imaging data of 92 hips from 61 children with treated DDH were retrospectively reviewed in Shandong Medical Imaging Research Institute from January 2019 to January 2020. All children underwent conservative treatments or surgical interventions 3 years ago. Hip function after treatment was evaluated clinically based on the modified MacKay criteria. The hips were divided into satisfactory clinical function group (McKay rating excellent or good, n=46) and unsatisfactory group (McKay rating fair or poor, n=46). All patients were imaged with conventional MRI, high resolution fat suppressed proton density weighted image (FS-PDWI) of the unilateral hip joint in oblique sagittal view, and anteroposterior hip radiographs. A-CAHI and lateral cartilaginous acetabulum-head-index (L-CAHI) were measured respectively on high-resolution oblique sagittal PDWI and conventional coronal T 1WI. Acetabulum head index (AHI) was also measured on anteroposterior hip radiograph. Mann-Whitney U test or independent-samples t test was used to compare the difference of A-CAHI, L-CAHI and AHI between satisfactory and unsatisfactory clinical function groups. The diagnostic value using A-CAHI, L-CAHI, AHI, or A-CAHI combined with L-CAHI for unsatisfactory clinical function were investigated by the ROC curve. The area under the curve (AUC) and the Z statistic were used to compare diagnostic performance. Results:The values of A-CAHI, L-CAHI and AHI were significantly higher in satisfactory clinical function group compared with the unsatisfactory group ( Z=-7.746, -7.735, t=-7.199, all P<0.001).A-CAHI combined with L-CAHI had the significant highest diagnostic accuracy compared with A-CAHI, L-CAHI and AHI (AUC were 0.994, 0.969, 0.968, 0.861, respectively), with significant differences ( Z=1.975, 2.006, 3.553, P=0.048, 0.051,<0.001). The sensitivity and specificity of A-CAHI combined with L-CAHI for the diagnosis of prognosis were 95.7% and 97.8%, respectively. Conclusions:A-CAHI measured by high resolution MRI was found to have the highest diagnostic accuracy for prediction of hip clinical function in the treated DDH, and combined with L-CAHI can improve the diagnostic accuracy significantly.

The application values of MR quantitative susceptibility mapping in differentiation between knee meniscus degeneration and tear / 中华放射学杂志

Objective:To evaluate the feasibility and the application values of quantitative susceptibility mapping (QSM) for the assessment of meniscal injury and in distinguishing meniscus degeneration and tears.Methods:The clinical and imaging data of 70 patients suspected of meniscus injury and scheduled for arthroscopy in Shandong Medical Imaging Research Institute, Cheeloo College of Medicine, Shandong University from November 2019 to June 2020 were analyzed retrospectively. Thirty age-and sex-matched healthy subjects were also examined as controls. All subjects received knee joint QSM and routine MR imaging. According to the results of arthroscopy, the patients was divided into meniscus degeneration and meniscus tear groups, respectively. The conventional MR was evaluated by two radiologists. The meniscus injury area was delineated on the original QSM magnitude images (the central area of the posterior corner of the lateral meniscus was selected in the healthy controls) and mapped to the corresponding QSM maps, and the magnetic susceptibility values were measured. Kruskal-Wallis H test was used to analyze the magnetic sensitivity values of meniscal degeneration, meniscal tear and healthy control groups; and Bonferroni was used to correct the pairwise comparison. ROC curve was established to evaluate the threshold and efficacy of magnetic susceptibility value in the diagnosis of meniscal tear. The results were compared with those of conventional MRI. Results:The magnetic susceptibility values of meniscus of healthy controls, meniscal degeneration and meniscal tear groups were (0.035±0.016)ppm, -0.031(-0.040,-0.005)ppm, and(-0.122±0.115)ppm, respectively, with significant difference found among the three groups (χ2=44.419, P<0.05). The magnetic susceptibility values of meniscus of healthy controls was significantly higher than those of meniscus degeneration patients and meniscus tear patients (χ2=-23.843, -48.253, P<0.05). The magnetic susceptibility values of meniscus of meniscus tear group was significant lower than those of meniscus degeneration group (χ2=-24.410, P<0.05). Taking magnetic susceptibility values of -0.062 5 ppm as threshold, the area under the ROC curve for the diagnosis of meniscal tears was 0.949, with the sensitivity as 87% and the specificity as 100%. The sensitivity and specificity of conventional MRI in the diagnosis of meniscal tears were 86.8% and 87.5%, respectively. Conclusion:QSM can quantitatively evaluate meniscus injury and can be used as an effective supplement method to conventional MRI, which is helpful to improve the diagnosis of meniscus tear.

Age-related study of brachial plexus MRI contrast to noise ratio and diffusion tensor imaging values in normal adults / 中华放射学杂志

Objective:To investigate the correlation of age with diffusion tensor imaging (DTI) values as fractional anisotropy (FA), apparent diffusion coefficient (ADC) and contrast to noise ratio (CNR) of three-dimensional nerve-sheath signal increased with inked rest-tissue rapid acquisition of relaxation imaging (3D SHINKEI) of the brachial plexus in normal adults.Methods:A total of 54 adult healthy volunteers and 6 patients with Guillain-Barre syndrome were prospectively enrolled from October 2018 to April 2019. Healthy volunteers were divided into 3 groups according to age as 21-40 years old group ( n=20), 41-60 years old group ( n=20), and ≥61 years old group ( n=14). All of them underwent MRI examination of the brachial plexus, including DTI and 3D SHINKEI sequences. The average FA and ADC values of the brachial plexus were measured and calculated through the fusion of DTI and 3D SHINKEI by 2 physicians independently. The contrast to noise ratio (CNR) of brachial plexus nerve was measured in 3D SHINKEI sequence images. Intraclass correlation efficient (ICC) was used to analyze the consistency between the two physicians. A simple linear regression model and Pearson correlation analysis were used to detect the correlation between FA, ADC, CNR and age. One-way analysis of variance was used to compare the differences of FA, ADC and CNR in different age groups. The FA and ADC values in different genders were compared by independent sample t-test. Results:Inter-observer agreements of the 2 physicians were good for FA and ADC values with ICC values of 0.811 and 0.901, respectively. For different groups, FA values were 0.397±0.023, 0.368±0.023, and 0.334±0.018 and ADC values were (1.376±0.072) × 10 -3 mm 2/s, (1.466±0.086) × 10 -3 mm 2/s, (1.486±0.080) × 10 -3 mm 2/s, for 21-40, 41-60, and ≥61 years old groups, respectively with statistical significant difference ( F=25.311, P<0.001; F=9.948, P<0.001). The CNR of the brachial plexus were 510.583±192.846, 502.581±128.821, and 426.782±113.648 for 21-40, 41-60, and ≥61 years old group without statistical difference ( F=1.429, P=0.249). The FA value of brachial plexus was highly negatively correlated with age ( r=-0.745, P<0.001), while the ADC value was moderately positively correlated with age ( r=0.596, P<0.001). The CNR of 3D SHINKEI sequence was negatively correlated with age ( r=-0.292, P=0.033). There was no statistically significant difference in brachial plexus FA and ADC values between male and female subjects ( t=1.496, P=0.141; t=-1.557, P=0.126). The FA value of Guillain-Barre syndrome patients was lower than that of healthy volanteers in the same age group ( t=6.129, P<0.001), and the ADC value had no statistical diference ( t=-1.335, P=0.186). Conclusion:The values of FA, ADC and CNR of brachial plexus in normal adults change with age. Among them, FA value is more significant.

Feasibility of prenatal MRI in the diagnosis of fetal external ear developmental malformation / 中华放射学杂志

Objective:To discuss the feasibility of prenatal MRI in evaluating fetal auricle developmental malformation and atresia of external auditory canal.Methods:Fifteen pregnant women (aged from 22 to 40 years old, mean age 31.3±5.2 years old) with fetal external ear developmental malformation suspected by ultrasound underwent MR scanning between November 2017 and May 2019. All of them were singleton. The gestational age ranged from 23 weeks to 35 weeks, with an average of (27.5±3.5) weeks. The sensitivity, specificity and accuracy of MRI and ultrasound in the diagnosis of fetal auricle malformation and atresia of external auditory canal were calculated and compared, using postnatal follow-up as the gold standard. Fisher exact test was used to compare the efficacy of MRI and ultrasound in diagnosing atresia of external auditory canal.Results:A total of 30 fetal external ears were detected in 15 fetuses, without auricle absence. Totally 19 external ears with developmental malformation were confirmed by postnatal follow-up, including 19 ears with auricle malformation and 15 ears with external auditory canal atresia. The accuracy of MRI and ultrasound in the diagnosis of auricle malformation was both 100% (19/19). For the diagnosis of external auditory canal atresia, the sensitivity, specificity and accuracy of MRI and ultrasound were 93.3% (14/15), 75.0% (3/4), 89.5% (17/19) and 33.3% (5/15), 25.0% (1/4), 31.6% (6/19), respectively. The sensitivity and accuracy of MRI in the diagnosis of atresia of external auditory canal were significantly higher than those of ultrasound, with statistically significant difference ( P=0.004, 0.001). Conclusion:MRI plays an important role in the diagnosis of fetal external ear developmental malformation, which can be used as an effective supplement to ultrasound, especially for the diagnosis of external auditory atresia.

Evaluation of the neurodevelopment outcome of prenatally diagnosed periventricular pseudocysts using MRI / 中华放射学杂志

Objective:To evaluate alterations of periventricular pseudocysts (PVPC) on MRI before and after birth, and to assess the prognosis.Methods:We retrospectively analyzed the data of 67 cases that were diagnosed with PVPC on prenatal MRI, of which 24 cases were lost to follow-up, 2 died after birth. A total of 41 surviving fetuses were included in this prognosis study. The gestational ages in this group were between 23 and 39 weeks, with an average of (33±3) weeks.All the subjects underwent brain MRI examinations and Gesell Developmental Scale (GDS) testing between 0-3 years of age. According to the location of cysts and with or without other intracranial and extracranial malformations (dilated ventricles orcerebella medulla, hypoxic-ischemic encephalopathy, TORCH virus infection, corporal hypoplasia, chromosomal malformations and nodular sclerosis) , the patients were divided into four groups: isolated connatal cysts, connatal cysts with additional findings,isolated subependymal pseudocysts, and subependymal pseudocysts with additional findings.The MR images were independently reviewed by two radiologists blinded to the clinical information. Intraclass correlation efficient (ICC) was used to analyze the consistency between the two reviewers.Chi-square test was used to compare the location of cysts (single/bilateral), the number of cyst cavities (single/multi-chamber), and other abnormalities in the connatal cyst group and subependymal cyst group. The mean anteroposterior diameter and mean height of cysts between the connatal cyst group and subependymal cyst group were compared by independent sample t-test.The ANOVA test was used to compare the differences in GDS outcomes among the groups. Multiple comparisons were conducted using the LSD test. Results:Inter-observer agreements between the two radiologists were good for the collected data (all ICC>0.75). Eleven isolated connatal cysts and 7 connatal cysts with additional findings became smaller or disappeared, and all had good prognosis. Of the 14 isolated subependymal cysts, 12 became smaller or disappeared, 2 had no change in size, and 13 had good prognosis. The subependymal cysts with additional findings group included 9 cases: 6 became smaller or disappeared, only 3 showed no apparent changes, and 7 had an abnormal outcome. Subependymal cysts with additional findings were significantly reduced and patients demonstrated significant differences compared with the those with isolated subependymal cysts in the development quotients (DQ) of adaptability, large movements, fine movements, personal social interaction, and language DQ ( P all<0.05). DQ between patients with isolated connatal cysts and isolated subependymal cysts was comparable ( P all>0.05). When associated with additional findings, connatal cysts and subependymal cysts could induce significant different DQ outcome ( P all<0.05). Conclusions:Isolated PVPC usually become smaller or disappeared and have a benign presentation after birth, whereas patients with subependymal cysts with additional findings usually have a poor prognosis. Connatal cysts usually have a good prognosis.

Re-examination of malaria diagnosis in Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019 / 中国血吸虫病防治杂志

ObjectiveTo analyze the re-examination results of malaria cases captured from the National Notifiable Communicable Disease Reporting System in Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, so as to pro- vide the scientific evidence for improving the malaria control capability in the province. MethodsMicroscopy and nested PCR assay were performed to re-examine the diagnosis of malaria cases registered in the National Notifiable Communicable Disease Reporting System in Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, and the coincidences of ma- laria diagnosis and malaria parasite species were evaluated. Results A total of 410 malaria cases were reported in Hubei Province from 2017 to 2019 according to the data retrieved from the National Notifiable Communicable Disease Reporting System. Among the 407 samples re-examined by Hubei Provincial Malaria Diagnostic Reference Laboratory from 2017 to 2019, the diag- nosis 374 malaria cases were confirmed, with an overall coincidence of 91.89% (374/407) for malaria diagnosis and 89.04% (333/374) for parasite species identification. The coincidence rates of malaria diagnosis and parasite species identification were 50.00% to 100.00% and 66.67% to 100.00% in 16 cities (prefectures) of Hubei Province during the re-examinations, which both varied in regions (χ2 = 40.46 and 42.30, both P values < 0.01). The coincidence rates of Plasmodium falciparum, P. vivax, P. malariae and P. ovale identification were 95.80%, 100.00%, 58.33% and 51.92% during the re-examinations, respectively (χ2 = 76.66, P < 0.01). The consistency rate between microscopic and nested PCR results was 89.83% (362/403). Conclusions The overall diagnostic quality of malaria is high in medical institutions at all levels in Hubei Province; however, the diagnostic capability of malaria remains to be improved in some regions.

Increased glycogen synthase kinase 3β activity involves in the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 in HT22 cells / 中华神经科杂志

Objective@#To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.@*Methods@#HT22 mouse hippocampal cells were divided into control group, Aβ31-35 group and LiCl+Aβ31-35 group by random number table method in the present study. Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)). Cell viability was detected by the cell counting kit-8 assay. The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times. The expression of GSK3β and BMAL1 protein was detected by Western blotting.@*Results@#Compared with the control group, Aβ31-35 induced the decreased expression of Bmal1 mRNA; The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA: 0.38±0.06 vs 0.83±0.08, t=4.549, P=0.001; BMAL1 protein: 0.67±0.04 vs 1.00±0.04, t=5.943, P<0.001). In the Aβ31-35 group, GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14, t=2.217, P=0.025). Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group, t=3.891, P=0.006), and the GSK3β inhibitor LiCl pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group, t=3.412, P=0.010). LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA: 0.72±0.05 vs 0.38±0.06, t=4.378, P=0.001; BMAL1 protein: 0.90±0.04 vs 0.67±0.04, t=4.052, P=0.002).@*Conclusion@#Increased GSK3β activity involved in the decreased expression of Bmal1 induced by Aβ31-35 in HT22 cells.

Increased glycogen synthase kinase 3β activity involves in the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 in HT22 cells / 中华神经科杂志

Objective@#To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.@*Methods@#HT22 mouse hippocampal cells were divided into control group, Aβ31-35 group and LiCl+Aβ 31-35 group by random number table method in the present study. Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)). Cell viability was detected by the cell counting kit-8 assay. The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times. The expression of GSK3β and BMAL1 protein was detected by Western blotting.@*Results@#Compared with the control group, Aβ31-35 induced the decreased expression of Bmal1 mRNA; The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA: 0.38±0.06 vs 0.83±0.08, t=4.549, P=0.001; BMAL1 protein: 0.67±0.04 vs 1.00±0.04, t=5.943, P<0.001). In the Aβ31-35 group, GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14, t=2.217, P=0.025). Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group, t=3.891, P=0.006), and the GSK3β inhibitor LiCl pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group, t=3.412, P=0.010). LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA: 0.72±0.05 vs 0.38±0.06, t=4.378, P=0.001; BMAL1 protein: 0.90±0.04 vs 0.67±0.04, t=4.052, P=0.002).@*Conclusion@#Increased GSK3β activity involved in the decreased expression of Bmal1 induced by Aβ31-35 in HT22 cells.

Increased glycogen synthase kinase 3β activity involves in the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 in HT22 cells / 中华神经科杂志

Objective To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.Methods HT22 mouse hippocampal cells were divided into control group,Aβ31-35 group and LiCl+Aβ 31-35 group by random number table method in the present study.Cells were synchronized to G0/G1 phase by 1％ serum starvation for 1 hour (circadian time 0 (CT0)).Cell viability was detected by the cell counting kit-8 assay.The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times.The expression of GSK3β and BMAL1 protein was detected by Western blotting.Results Compared with the control group,Aβ31-35 induced the decreased expression of Bmal1 mRNA;The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA:0.38±0.06 vs 0.83±0.08,t=4.549,P=0.001;BMAL1 protein:0.67±0.04 vs 1.00±0.04,t=5.943,P＜0.001).In the Aβ31-35group,GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14,t=2.217,P=0.025).Aβ31-35 decreased the viability of HT22 cells (71.85％±6.20％ in the Aβ31-35 group vs 98.14％±2.68％ in the control group,t=3.891,P=0.006),and the GSK3β inhibitor LiC1 pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74％±5.74％ in the LiCl+Aβ31-35 group vs 71.85％±6.20％ in the Aβ31-35 group,t=3.412,P=0.010).LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA:0.72±0.05 vs 0.38±0.06,t=4.378,P=0.001;BMAL1 protein:0.90±0.04 vs 0.67±0.04,t=4.052,P=0.002).Conclusion Increased GSK3β activity involved in the decreased expression of Bmal 1 induced by Aβ31-35 in HT22 cells.

Effect of Fengshi Qutong Capsule on Human Umbilical Vein Endothelial Cells Induced by VEGF / 中国实验方剂学杂志

Objective: To study the effect of Fengshi Qutong capsule on the migration, adhesion, invasion and tube formation of human synovial cells and the phosphorylation and protein expression of vascular endothelial growth factor receptor 2 (VEGFR2). Method: With human umbilical vein endothelial cells (HUVEC) as the research object, low, middle and high-dose Fengshi Qutong capsule(0.02,0.1,0.5 μg·L-1) on HUVEC was determined by methye thiazolye telrazlium (MTT) colorimetric assay for the follow-up experiment. The transwell migration, adhesion and transwell invasion test were used to detect the migration and adhesion of the different concentrations of Fengshi Qutong capsule in HUVEC. The expression of VEGFR2 in HUVEC was detected by Western blot, and Real-time PCR was used to detect the content of VEGFR2 mRNA in cells. Result: Compared with normal group, the proliferation of HUVEC was significantly increased after 24 h and 48 h of VEGF induction (PPP-1 Fengshi Qutong capsule were administered in vitro for 48 h to inhibit HUVEC proliferation activity in a dose-dependent manner (PPPPConclusion: Fengshi Qutong capsule can inhibit the migration, adhesion, invasion and tube formation of HUVEC. This effect may be related to the inhibition of phosphorylation, and protein and mRNA expression level of VEGFR2.

Effect of Fengshi Qutong Capsule on Synovial Akt and MAPK Signaling Pathways in Rheumatoid Arthritis / 中国实验方剂学杂志

Objective:To observe the effect of Fengshi Qutong capsule (FSQTC) on protein kinase B(Akt) and mitogen-activated protein kinase (MAPK) signaling pathways of rheumatoid arthritis (RA). Method:Collagen-induced arthritis (CIA) was induced in SD rats, and the synovial membranes of the knee joints were prepared after 19 days of oral administration of 0.25, 0.5, 1 g·kg-1 FSQTC. MH7A cells were induced by tumor necrosis factor-α (TNF-α, 20 μg·L-1) in vitro, and human umbilical vein endothelial cells (HUVEC) were induced by vascular endothelial growth factor (VEGF). FSQTC (0.02, 0.1, 0.5 μg·L-1) were added to MH7A/HUVEC cells, and then the cells were collected. Proteins of synovial tissue, MH7A and HUVEC cells were extracted, and then were detected the expresstion of p-Akt, p-p38 MAPK, p-extracellular signal-regulated kinase(ERK) and p-Jun n-terminal kinase(JNK) by Western blot. Result:The expression levels of p-Akt, p-p38 MAPK, p-ERK and p-JNK in the synovial membrane of CIA model were significantly increased compared with normal group (P-1·d-1 FSQTC significantly decreased their expression levels (PPα or VEGF were increased (P-1 FSQTC (PPConclusion:FSQTC can down-regulate the abnormal activation of Akt and MAPK signaling pathways in the synovial membrane of CIA rats, fibroblast synovial cells and vascular endothelial cells, which is related to the inhibition of synovial angiogenesis in the treatment of RA.

Effect of Fengshi Qutong Capsule on Function of Human Synovial Cells in Rheumatoid Arthritis Induced by TNF-α / 中国实验方剂学杂志

Objective: To study the effects of Fengshi Qutong capsule (FSQTC) on proliferation, migration, adhesion, invasion and secretion of human synovial cells in rheumatoid arthritis (RA) induced by tumor necrosis factor-α (TNF-α) and explore its mechanism. Method: Human synovial cells (MH7A) in RA patients were induced in vitro by using TNF-α (20 μg·L-1). After treatment with different concentrations of FSQTC (0.02,0.1,0.5 μg·L-1), MTT colorimetric assay, transwell migration, adhesion and invasion tests were used to detect the proliferation, migration, adhesion and invasion of the MH7A, respectively. The expression levels of vascular endothelial growth factor (VEGF) and interleukin-1β (IL-1β) in MH7A supernatant were detected by enzyme linked immunosorbent assay (ELISA). Result: As compared with blank control group, TNF-α (20 μg·L-1) significantly increased the proliferation, migration, adhesion, invasion and secretion of IL-1β and VEGF of MH7A cells (P-1) had no significant effect on proliferation of TNF-α-induced MH7A cells after treatment for 24 hours. After 48 hours of treatment, proliferation of MH7A cells induced by TNF-α was decreased in a concentration-dependent manner (PPPPConclusion: FSQTC can inhibit the proliferation, migration, adhesion, invasion and secretion of IL-1β and VEGF in MH7A cells.

Effect of Fengshi Qutong Capsule on Collagen-induced Arthritis of Rats / 中国实验方剂学杂志

Objective:To observe the intervention effect of Fengshi Qutong capsule on collagen-induced arthritis (CIA) in rats. Method:SD rats were randomly divided into normal group, model group, low, medium and high-dose Fengshi Qutong capsule groups (0.25, 0.5, 1 g·kg-1·d-1), and methotrexate group(0.2 mg·kg-1·d-1).Except for normal group, the other groups were immunized with type Ⅱ collagen and incomplete Freund's adjuvant to establish a CIA model. On the 1st day after the first immunization, the administration group was given intragastric administration, once a day, for 19 days; on the 8th day after the first immunization, the symptoms of joint swelling and malformation of the rats were observed, and the clinical scores and incidence of arthritis were evaluated. On the 19th day, micro-computed tomography and bone metrology were performed, and histopathological examination of inflammatory joints was performed,andsynovial inflammation,vasospasm, cartilage erosion and bone destruction by pathological severity scores, serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF),matrix metalloproteinase-3 (MMP-3) and receptor activator of nuclear factor kappa B ligand(RANKL)were detected by enzyme linked immunosorbent assay. Result:Fengshi Qutong capsule could improve the symptoms of inflammatory joint redness, swelling and deformity in CIA rats in a dose-dependent manner. Compared with normal group, clinical score and incidence, joint synovial inflammation, vasospasm, cartilage erosion and pathological score of bone destruction in joint group were significantly increased (PPPβ, TNF-α, VEGF, MMP-3 and RANKL in serum were increased (PPPPPPPβ, TNF-α, VEGF, MMP-3 and RANKL were significantly decreased (PPConclusion:Fengshi Qutong capsule can effectively alleviate the clinical symptoms and conditions of experimental rheumatoid arthritis in rats, reduce the incidence, and relieve the histopathology and imaging severity, while inhibiting the inflammatory cytokines.

Effect of Fengshi Qutong Capsules on synovial angiogenesis in rats with type Ⅱ collagen induced arthritis / 中国中药杂志

To observe the effect of Fengshi Qutong Capsules(FSQTC) on angiogenesis of rat aortarings and in knee joint synovium of type Ⅱ collagen-induced arthritis(CIA) rats. The blood vessel of aorta rings of normal SD rats were induced by vascular endothelial growth factor(VEGF) 20 μg·L~(-1 )in vitro, and were treated with FSQTC(0.02, 0.1 and 0.5 μg·L~(-1)) continuously for 9 days. The number, length and area of neovascularization of the vascular ring were measured. SD rats were immunized to establish collagen-induced arthritis. CIA rats were treated with FSQTC(0.25, 0.5, 1 g·kg~(-1)·d~(-1)) and methotrexate(0.2 mg·kg~(-1)·d~(-1)) daily for 19 days. Histopathological examination(HE) was performed to observe the vascular morphology and vascular density in the synovial membrane of the inflamed joint. Immunohistochemistry was performed to observe the expression of platelets-endothelial cell adhesion molecule(CD31), VEGF and VEGF receptor 2(VEGFR_2)in the synovium. Immunofluorescence was performed to observe the expression of CD31 and α smooth muscle actin(αSMA) in synovial membrane.TGF-β, PDGF and VEGFR_2 in serum were detected by enzyme-linked immunosorbent assay. The number, branch length and area of blood vessels of aorta rings were significantly increased induced by VEGF, and FSQTC could significantly reduce the number, branch length and area of blood vessels. Compared with the normal group, the vascular density, CD31 positive expression, CD31~+/αSMA~- immature and total vascular positive expression in the synovial membrane of the model group were significantly increased, and so as VEGF and VEGFR_2 in the synovium. The VEGFR_2, TGF-β and PDGF in sera were also significantly increased in model group. FSQTC reduced the synovial vascular density and inhibited the positive expression of CD31, CD31~+/αSMA~- immature blood vessels and total vascular. FSQTC has no significant effect on CD31~+/αSMA~+mature blood vessels. FSQTC also negatively inhibited the expression of VEGF, VEGFR_2, TGF-β and PDGF in synovial membrane and/or sera. The effect of methotrexate is similar with to the high dose group. Our results demonstrated that FSQTC could inhibit the angiogenesis of synovial tissue in CIA rats and of aortaring in rats, which is related to the reduction of angiogenesis regulatory factor.

Susceptibility weighted imaging in evaluation of the fetal vertebra and vertebral anomalies / 中华放射学杂志

Objective To explore the advantages of susceptibility weighted imaging (SWI) in depiction of normal fetal vertebra and vertebral anomalies.Methods This prospective study was approved by our institutional review board, and written informed consent was obtained from every participant, Fifty-eight pregnant women (gestation age 22 to 39 weeks, average 29 ± 3 weeks) who were suspected of carrying babies with vertebral anomalies by ultrasound screening underwent 1.5 T fetal spine MRI[including half-fourier acquisition single-shot turbo spin-echo(HASTE),true fast imaging with steady-state(True FISP) and SWI sequences]. MR images were reviewed for their quality by two radiologists independently. The image scores in HASTE, True FISP and SWI were compared by using Kruskal-Wallis test and Mann-Whitney U test. Three segments (cervical, thoracic and lumbosacral segments, respectively) of 15 fetuses were, at random, collected to compare among HASTE,True FISP and SWI and then evaluated by ANOVA analysis.The diagnostic accuracy of the three sequences among 32 cases with follow-up results was calculated respectively and compared by using Chi-square test. Results There was statistical differences among three sequences(χ2=50.685,P<0.05).The scores of SWI was higher than that of True FISP, and the scores of True FISP was higher than that of HASTE in the evaluation of the fetal vertebra(P all<0.05).The differences among cervical,thoracic and lumbosacral segments on True FISP and HASTE showed significant difference statistically (P all<0.05), also the image quality of cervical segment could not meet the requirement of clinical diagnosis. The image quality of SWI was favorable clinically and no statistical difference among three segments was found(P>0.05).A total of 32 fetal vertebral anomalies were identified by follow-up after birth including hemivertebra (n=14), fusion of vertebrae (n=1), butterfly vertebra (n=1), multiple vertebral malformations(n=9),spinal bifida(n=5),caudal regression syndrome(n=2).The diagnostic accuracy of SWI, True FISP and HASTE was 93.75% (30/32), 56.25% (18/32) and 37.50% (12/32) respectively.The diagnostic accuracy of SWI was the best compared to that of True-FISP and HASTE(χ2=10.083,20.017;P<0.01). Conclusion SWI proved to be the optimal technique in depiction of fetal vertebra and vertebral anomalies than True FISP and HASTE,especially in depiction of cervical vertebra.

Deubiquitinases as pivotal regulators of T cell functions / 医学前沿

T cells efficiently respond to foreign antigens to mediate immune responses against infections but are tolerant to self-tissues. Defect in T cell activation is associated with severe immune deficiencies, whereas aberrant T cell activation contributes to the pathogenesis of diverse autoimmune and inflammatory diseases. An emerging mechanism that regulates T cell activation and tolerance is ubiquitination, a reversible process of protein modification that is counter-regulated by ubiquitinating enzymes and deubiquitinases (DUBs). DUBs are isopeptidases that cleave polyubiquitin chains and remove ubiquitin from target proteins, thereby controlling the magnitude and duration of ubiquitin signaling. It is now well recognized that DUBs are crucial regulators of T cell responses and serve as potential therapeutic targets for manipulating immune responses in the treatment of immunological disorders and cancer. This review will discuss the recent progresses regarding the functions of DUBs in T cells.