Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.

Objective:To analyze any effect of combining hyperbaric oxygen with qingkailing on the metabolic biomarkers of consciousness disorder (CD) caused by brain trauma using metabolomics technology. Also, to screen out diagnostic markers for clinical treatment and prognosis.Methods:Thirty-six patients with a CD caused by brain trauma formed the observation group, while 40 healthy gender-, age- and ethnicity-matched individuals were the control group. Both groups were given routine supportive treatment, while the observation group additionally received hyperbaric oxygen and oral qingkailing medication. Before and after 14 days consciousness disturbance was evaluated using the Glasgow Coma Scale (GCS). Venous blood was collected for metabolomic analysis using liquid chromatography linked to a mass spectrometer to screen out specific metabolic biomarkers. Metabolic markers associated with the disease and of diagnostic significance were thus identified.Results:After the treatment the average GCS score of the observation group had improved significantly and the degree of coma was significantly relieved. Twenty-one metabolic markers were found to be significant, with creatine, proline, uric acid, acetyl-L-carnitine, histidine, proline leucine, tryptophan and 9E-octagenoic acid potentially of high diagnostic value. After the treatment, all of those markers came close to the levels observed in the healthy control group.Conclusions:Proline, leucine, 9E-octagenoleic acid, uric acid and acetyl-L-carnitine could be used for diagnosis and evaluating efficacy with such patients. Hyperbaric oxygen supplemented with qingkailing can relieve coma, correct metabolic disorders and accelerate patients′ awakening. Metabolomics provides a new method for identifying endogenous metabolic abnormalities in patients with post-traumatic consciousness disorders. It can be useful in prognosis and clinical treatment.

AIM:To study the effect of NEDD8-conjugating enzyme UBE2F on lung adenocarcino-ma metastasis.METHODS:The expression of UBE2F in lung adenocarcinoma was analyzed using TIMER2.0,UALCAN and HPA databases.Kaplan-Mei-er Plotter database was used to analyze the rela-tionship between UBE2F expression and survival rate of lung adenocarcinoma.A UBE2F-knockout lung adenocarcinoma cell line was constructed us-ing CRISPR/Cas9 technology,and a UBE2F-knockout lung adenocarcinoma metastasis model was con-structed in nude mice to verify the effect of UBE2F knockout on lung adenocarcinoma metastasis.The effects of UBE2F knockout on invasion and migra-tion of lung adenocarcinoma cells were examined by cell scratch assay and Transwell invasion and mi-gration assays.The effect of down-regulated UBE2F expression on snail expression,a key marker of epi-thelial-mesenchymal transition(EMT),was detect-ed by Western blot and Real time PCR.RESULTS:Multiple database analysis showed that UBE2F was highly expressed in lung cancer,and Kaplan-Meier Plotter analysis showed that high expression of UBE2F in lung adenocarcinoma had better progno-sis than low expression.In vivo experiments showed that compared with control group,the number of nodules metastasized on the lung sur-face of nude mice after UBE2F knockout was signifi-cantly increased(P<0.05).Cell scratch assay and Transwell assay showed that UBE2F enhanced the migration and invasion ability of lung cancer cells after knockout,and the difference were statistically significant(P<0.05).Western blot and Real time PCR results indicated that the level of EMT tran-scription factor snail protein and mRNA increased after UBE2F knockout.CONCLUSION:In lung ade-nocarcinoma cells,UBE2F down-regulation leads to Snail accumulation and promotes invasion and me-tastasis of lung adenocarcinoma cells.

Abstract Astragalus polysaccharides(APS), as one of the main active components of Astragali Radix, a traditional famous tonic drug in China, has good pharmacological effects in regulating immunity and anti-oxidation. In recent years, with the success of PD-1/PD-L1 inhibitors and anti-CTLA-4 drugs in cancer treatment, immunotherapy has truly become one of the main clinical anti-tumor treatment methods. Aiming at the characteristics of tumor immune escape and tumor microenvironment change, this paper explores the clinical potential of APS as an anti-tumor adjuvant drug, and summarizes the role and mechanism of APS in inhibiting tumor immune escape, regulating tumor microenvironment, and improving immune function. To summarize and analyze the immune regulation of APS on various tumors in order to provide an objective basis for APS as a clinical anti-tumor adjuvant drug.

OBJECTIVE To establish a quantitative analysis method for the quality control components in Tenghuang jiangu capsules， and predict the possible action mechanism of the quality control components. METHODS Seven key quality control components in Tenghuang jiangu capsules were quantitatively analyzed by UHPLC-Q-Orbitrap HRMS. The “component-target” network was constructed based on network pharmacology， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and gene ontology （GO） function enrichment analysis were further conducted to find the key signaling pathways. RESULTS The average contents of succinic acid， hyperoside， gallic acid， kaempferol， naringin， naringenin and protocatechuic acid in 20 batches of Tenghuang jiangu capsules were 520.92， 67.67， 129.48， 4.74， 397.45， 5.66 and 376.62 μg/g， respectively. The results of network pharmacology showed that the 62 key target genes of the quality control components of the drug included AKT1， TNF， VEGFA， MMP9， PTGS2， etc. They were mainly enriched in cytokine receptor interaction， nuclear factor， tumor necrosis factor， interleukin 17， rheumatoid arthritis， Toll-like receptor and other signal pathways， involving inflammatory reaction， signal transduction， protein phosphorylation and other biological processes， kytoplasm， cell membrane and other cell components， as well as enzyme activity， energy activity and other molecular functions. CONCLUSIONS The established UHPLC- Q-Orbitrap HRMS method can be used for the quantitative analysis of the quality control components of Tenghuang jiangu capsule. Its quality control components may be mapped to inflammatory pathways related to bone diseases such as rheumatoid arthritis and Toll-like receptors through AKT1， TNF， VEGFA and other key targets， so as to play a therapeutic role.

Objective:To investigate the abnormal patterns of spontaneous neural activity of patients with type 2 diabetes comorbid depression (T2DD) by using resting-state functional magnetic resonance imaging (rs-fMRI) fractional amplitude of low-frequency (fALFF) analysis, and determine the neuroimaging features of brain damage in T2DD patients.Methods:A perspective study was performed. Fifty-nine type 2 diabetes mellitus (T2DM) patients and 52 T2DD patients, admitted to and accepted treatment in Department of Endocrinology of our hospital from November 2017 to November 2020, were chosen; another 57 healthy controls matched with gender, age and education level, admitted to our hospital at the same time period were enrolled. Their clinical data, neuropsychological test and rs-fMRI data were collected; whole brain fALFF values were calculated, and fALFF values of different brain regions were compared in subjects of the 3 groups. Pearson correlation analysis was used to verify the correlations of fALFF values with clinical variables and neuropsychological scale scores.Results:The fALFF values in bilateral precuneus showed significant difference among the three groups ( P<0.05). The fALFF values in bilateral precuneus of the T2DD and T2DM groups were significantly lower than those in heathy control group ( P<0.05), and those in the T2DD group were lower than those in the T2DM group without significant difference ( P>0.05). Pearson correlation analysis showed that there were no correlations of fALFF values with clinical data and psychometric scale scores in T2DD group and T2DM group ( P>0.05). Conclusion:The abnormal patterns of spontaneous brain activity in the bilateral precuneus may be the neuroimaging markers of brain damage in T2DD patients.

Objective: To investigate the alterations of the cerebral resting-state spontaneous neural activity in colorectal cancer patients with depressive symptoms. Methods: Thirty-three colorectal cancer patients (patient group) with depression and 43 healthy subjects (control group) underwent the resting state functional magnetic resonance imaging (rs-fMRI). The amplitude of low-frequency fluctuations (ALFF) and fractional ALFF (fALFF) were calculated. Two independent samples t test were used to compare the ALFF and fALFF values between two groups by DPABI software, and then correlation analysis was performed between ALFF and fALFF with statistical significance and Patient Health Questionnaire (PHQ-9) scores and Generalized Anxiety Disorder (GAD-7) scores. Results: Compared with the control group, the patient group showed significantly lower ALFF and fALFF values in the bilateral precuneus, calcarine gyrus, lingual gyrus, left cuneus, superior, middle, inferior occipital gyrus and right fusiform gyrus (t=-5.730, P<0.05; t=-4.872, P<0.05). There were no significant correlations between the ALFF and fALFF values in these regions and PHQ-9 or GAD-7 scores (P>0.05). Conclusion Spontaneous decrease of neural activity in occipital and parietal lobes exists in colorectal cancer patients with depression at resting-sate, which may be a potential neurobiological marker.

Objective? To investigate the status of caregiver preparedness and adult attachment among the spouse of young and middle-aged stroke patients,and to analyze the correlation between them. Methods? From January 2017 to May 2018,110 spouses of stroke patients who were treated in Affiliated Hangzhou First People＇s Hospital, Zhejiang University School of Medicine were selected as subjects by convenience sampling. The General Information Questionnaire,Care Preparedness Scale (CPS),and Experience in Close Relationship Scale (ECR) were used in the survey. Totally 110 questionnaires were distributed, and 102 valid ones were retrieved, yielding an effective recovery rate of 92.73%. Results? The total CPS score of 102 young and middle-aged stroke patients was (12.87±5.39). The scores of attachment anxiety and attachment avoidance in ECR were (57.96 ±11.69) and (48.06 ±12.10). There were statistically significant differences in the scores of preparedness of caregivers among spouse caregivers of different ages, self reported health status and nursing experience (P＜ 0.05). Attachment anxiety dimension was negatively correlated with caregiver preparedness score (r=-0.318,P＜0.01), and attachment avoidance dimension was negatively correlated with caregiver preparedness score (r=-0.233,P＜ 0.01). Conclusions? The preparedness status of the spouses caregivers of young and middle-aged stroke patients are at a low level. Medical workers can improve their care preparedness by giving them home care knowledge guidance,reducing their attachment avoidance and attachment anxiety so as to improve their preparedness.

Objective To investigate the changes of microstructure and blood perfusion of thalamus and basal ganglia in patients with untreated depressive disorder using diffusion kurtosis imaging (DKI) and three-dimensional arterial spin labeling (3D ASL) perfusion imaging.Methods Totally 21 untreated patients with depressive disorder (disease group) and 32 healthy volunteers (control group) underwent DKI,3D ASL and routine MR scanning were enrolled.DKI parameters (mean kurtosis [MK],axial kurtosis [Ka],radial kurtosis [Kr],fractional anisotropy [FA],mean diffusivity [MD],axial diffusivity [Da] and radial diffusivity [Dr]) and 3D ASL parameters (cerebral blood flow [CBF]) of bilateral heads of caudate nucleus,thalamus and putamens were measured and compared between the 2 groups.Results Compared with control group,Ka of left caudate head in disease group was significantly higher,while Kr was significantly lower (both P＜ 0.05).Ka of right putamens in disease group was significantly higher than that in control group (P＜0.05).Compared with control group,CBF of left caudate head and bilateral putamens decreased significantly in disease group (all P＜0.05).Conclusion Untreated depressed patients show abnormal DKI parameters and blood perfusion in basal ganglia,which indicating that microstructure integrity is impaired and blood perfusion decreases.The abnormalities in basal ganglia microstructure and blood perfusion may serve as one of the neuroimaging features of depressive disorders.

OBJECTIVE: Bipolar disorder (BD) is often misdiagnosed as unipolar depression (UD), leading to mistreatment and poor clinical outcomes. However, little is known about the similarities and differences in subcorticalgray matter regions between BD and UD. METHODS: Thirty-five BD patients, 30 UD patients and 40 healthy controls underwent diffusional kurtosis imaging (DKI) and three dimensional arterial spin labeling (3D ASL). The parameters including mean kurtosis (MK), axial kurtosis (Ka), radial kurtosis (Kr), fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (Da), radial diffusivity (Dr) and cerebral blood flow (CBF) were measured by using regions-of-interest analysis in the caudate, putamen and thalamus of the subcortical gray matter regions. RESULTS: UD exhibited differences from controls for DKI measures and CBF in the left putamen and caudate. BD showed differences from controls for DKI measures in the left caudate. Additionally, BD showed lower Ka in right putamen, higher MD in right caudate compared with UD. Receiver operating characteristic analysis revealed the Kr of left caudate had the highest predictive power for distinguishing UD from controls. CONCLUSION: The two disorders may have overlaps in microstructural abnormality in basal ganglia. The change of caudate may serve as a potential biomarker for UD.
Anisotropy ; Basal Ganglia* ; Bipolar Disorder ; Cerebrovascular Circulation ; Depressive Disorder ; Diffusion* ; Gray Matter ; Humans ; Perfusion Imaging* ; Perfusion* ; Putamen ; ROC Curve ; Thalamus*