Adult ; Aged ; Aqueous Humor ; virology ; Female ; Humans ; Immunocompetence ; immunology ; Inflammation ; virology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective:To explore the mechanism of Sailuotong capsules in treating acute cerebral ischemia from the perspective of metabonomics. Method:A total of 24 SD rats were randomly divided into 3 groups, including sham-operated group, model group and Sailuotong group (33 mg·kg^-1). The rat model of acute multiple cerebral infarction was established by injecting fluorescent microspheres into internal carotid artery. After the successful operation, rats in Sailuotong group were administered by duodenal injection immediately, and the dosage volume was 2 mL·kg^-1. Endogenous metabolites in rat brain tissues of each group were determined by UPLC-Q-TOF-MS. The relevant data and biomarkers were analyzed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Result:The analysis of pattern recognition indicated that the metabolite profiles in model group and sham-operated group were separated obviously, and ten biomarkers related to acute cerebral ischemia were also identified. Compared with the sham-operated group, contents of N-acetylaspartate (NAA), fumaric acid, glutathione, dehydroascorbic acid, aspartic acid and S-adenosylhomocysteine were decreased, while the contents of arginine, citrulline, saccharopine and hydantoin-5-propionic acid were increased in the model group. Meanwhile, the ten abnormal biomarkers mentioned above got restoration in Sailuotong group. Conclusion:The main regulated metabolic pathways of Sailuotong capsules are NAA metabolism, arginine metabolism, energy metabolism, oxidative stress, etc.

Objective: To evaluate the utility of the BRAF^V600E mutation detection in different cytoloy categories and to determine if the VE1 antibody can serve as a screening tool for the detection of BRAF^V600E mutation in thyroid fine needle aspiration (FNA) specimens. Methods: A total of 273 FNA residual specimens were collected. BRAF^V600E testing was performed on these liquid-based specimens. And also 78 specimens with enough residual cells were stained with VE1 antibody. Comparisons of molecular and immunocytochemistry results with clinicopathological outcomes were performed. SPSS 17.0 software was used to analyze the data. Results: There were 70 indeterminated diagnoses in 273 cases with FNAs. Fifty-eight cases were proven to be papillary thyroid carcinoma (PTC) by histology, including 9 cases of atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS), 3 cases of follicular neoplasm/suspicious for follicular neoplasm (FN/SFN), and 46 cases of suspicious for malignancy (SM). BRAF^V600E analysis detected PTC in 3 of 9 cases with AUS/FLUS, and in 31 (67.4%) of 46 cases with SM. The sensitivity of immunostaining with VE1 antibody was 62.8%(27/43) and the specificity was 91.4% (32/35). VE1 expression showed moderately concordance with the molecular mutation (κ=0.524, P<0.001). Conclusions Additional BRAF^V600E detecting can improve the diagnostic efficacy for PTC in AUS/FLUS and SM. VE1 expression may be an alternative method for BRAF^V600E detecting when molecular detecting is unavailable.

Objective: To explore the feasibility of bronchoscopic brushing liquid-based slide cytology combined with automatic immunocytochemistry (ICC) for pathological typing of lung cancer. Methods: A liquid-based thin-prep was prepared from 171 bronchoscopic brushing specimens of patients with pulmonary lesions. ICC was detected by automatic immunohistochemistry instrument while cytomorphological diagnosis was made. The results were compared with those of histopathological diagnosis. Results: Among 171 patients, 130 (76.0%) could be classified by cell morphology alone, including 31 squamous cell carcinomas, 44 adenocarcinomas and 55 small cell carcinomas; 162 (94.7%) could be classified by cell morphology combined with ICC, including 38 squamous cell carcinomas, 61 adenocarcinomas and 63 small cell carcinomas (P<0.001). According to the gold standard of histopathological diagnosis, the coincidence rate of cytomorphology combined with ICC was higher than that of cell morphology alone. The coincidence rate of squamous cell carcinoma was increased from 85.2% to 97.1% (P=0.093), adenocarcinoma from 92.5% to 98.0% (P<0.001), and small cell carcinoma from 96.1% to 98.3% (P=0.465). Conclusion The combination of liquid-based thin-prep cytology and automatic immunohistochemistry can effectively improve the accuracy of pathological typing of brushing specimens under fiberoptic bronchoscopy, and provide more objective diagnostic results for clinical treatment.

Objective To explore developmental states on the preterm and term neonatal brain white matter based on the co-variation of metrics derived from diffusion tensor imaging(DTI).Methods This work enrolled 66 neonates,consisting of 33 preterm(21 males and 12 females;gestational age:30.143-36.286 weeks)and 33 full-term neonates(22 males and 11 females;gestational age:37.000 -41.429 weeks).DTI derived metrics included axial diffusivity(AD),radial diffusivity(RD),and fractional anisotropy(FA).Spearman's rank correlation between metrics and postmenstrual age was analyzed in the preterm and term neonates.Developmental states were evaluated according to the method proposed by Dubois:fiber organization was associated with increased AD, decreased RD and increased FA;pre-myelination was related to decreased AD,decreased RD and unchanged/increased FA;myelination was revealed by unchanged AD,decreased RD and increased FA.Mann-Whitney U test was used to compare DTI metrics between the preterm and term neonates.Results According to the method proposed by Dubois,posterior limb of internal capsule and corpus callosum underwent myelination in the preterm-neonate period.Cerebral peduncle started myelination in the term-neonate period.Superior corona radiate,inferior fronto-occipital fasciculus and external capsule underwent pre-myelination on preterm and term neonates.FA values were higher in term neonates than those in preterm neonates in all the selected regions(P＜0.05).AD and RD values were lower in term neonates than those in preterm neonates in the selected regions except for cerebral peduncle(P＜0.05).Conclusion Changes in DTI metrics and the method of Dubois can be used to quantitatively evaluate developmental states of the neonatal brain white matter.The changes coincided with DTI variations associated with pre-myelination and myelination.White matter development is delayed in preterm neonates compared with term neonates.

Objective To investigate the reliability of MRI classification and clinical significance of deep gray matter injury(DGMI)in children with cerebral palsy(CP).Methods We made a retrospective assessment of 14 children with gross motor function classification system(GMFCS),manual ability classification system(MACS)and MRI classification system of deep gray matter injury.Based on T2WI,two radiologists worked independently and graded MRI pictures according to three-grading system and four-grading system.To evaluate the reliability of different grading systems,intra-observer and inter-observer agreements were tested by Kappa test.Spearman correlation analysis was performed to analyze the MRI classification system with GMFCS and MACS.Results The Kappa value of the intro-observer and inter-observe agreement of three-grading system was 0.873 and 0.873,respectively (P＜0.001).The Kappa value of the intro-observer and inter-observe agreement of four-grading system was 0.901 and 0.611(P＜0.001).Three-grading system had no significant correlation with GMFCS(r=0.053,P＞0.05)or MACS(r=0.128,P＞0.05).Four-grading system had a significant positive correlation with GMFCS(r=0.605, P＜0.05)and MACS(r=0.779,P＜0.05).Conclusion In the two grading systems,four-grading system is a more repeatable approach for detecting deep gray matter,gross motor function and manual function injuries in children with cerebral palsy.

OBJECTIVETo explore the role of the interaction between glycogen synthase kinase-3β (GSK-3β) and endoplasmic reticulum stress (ERS) in the high glucose (HG)-induced injury in human umbilical vein endothelial cells (HUVECs).

METHODSHUVECs treated with 40 mmol/L glucose for 24 h were examined for expression levels of GSK-3β, GRP78, CHOP and cleaved caspase-3 protein using Western blotting. The cell viability was examined using CCK-8 assay and cell apoptosis was detected with Hoechst 33258 nuclear staining and photofluorography. The intracellular level of reactive oxygen species (ROS) was measured with dichlorfluoresein staining and photofluorography. Mitochondrial membrane potential (MMP) was tested by rhodamine 123 (Rh123) staining and photofluorography.

RESULTSTreatment of HUVECs with 40 µmol/L glucose for 3-24 h activated GSK-3β in a time-dependent manner, leading to significantly down-regulated expression of phosphorylated (p)-GSK-3β (P<0.05). HG exposure of the cells for 1-24 h induced ERS, evidenced by time-dependently up-regulated expression of GRP78 and CHOP (P<0.05). LiCl, an inhibitor of GSK-3β, attenuated HG-induced ERS and significantly lowered the expression levels of GRP78 and CHOP (P<0.01). 4-PBA, an inhibitor of ERS, obviously ameliorated the activation of GSK-3β by HG as shown by the increase in p-GSK-3β expression level (P<0.01). HG exposure for 24 h induced obvious injuries in HUVECs, which exhibited decreased cell viability, increased cell apoptosis, increased expression of cleaved caspase-3 and ROS generation, and loss of MMP. Pretreatment of the cells with LiCl or 4-PBA for 60 min before HG exposure significantly lessened the cell injuries (P<0.01).

CONCLUSIONInteractions between GSK-3β and ERS occur in HUVECs exposed to HG and participate in HG-induced cell injuries.

The fragmentation pathways of the three ginkgolides (ginkgolides A, ginkgolides B, ginkgolides C) have been studied with high resolution and high mass accuracy using quadrupole time-of-flight mass spectrometry in negative ion mode in this paper. The results indicate that the three ginkgolides have similar fragmentation pathways, including four kinds of common cleavage pathways and one common characteristic ion. In high quality regions, the typical fragmentation pathways of the three ginkgolides are lactone ring opening with continuous loss of CO, CO₂,and loss of H₂O. In low quality regions, the common characteristic fragment ion of the three ginkgolides at 72.993 6 is formed by C rings cleavage. Also, the common fragment ions of ginkgolides A and ginkgolides B at 141.018 8, 125.023 8, 113.024 0, 97.029 1 are formed by A rings cleavage. The study of fragmentation pathways could be adopted for the structural identification of the ginkgolides and their metabolites.
Ginkgolides ; chemistry ; Mass Spectrometry ; Spectrometry, Mass, Electrospray Ionization

PURPOSE: We have found that expression of γδT cells is increased in pathological mucosa of chronic rhinosinusitis with nasal polyps (CRSwNP) compared with normal nasal mucosa. This increase is correlated with the infiltration of eosinophils in CRSwNP. Here, we investigated the expression of γδT cells, inflammation and tissue remodeling factors as well as their probable relationships in different types of chronic rhinosinusitis (CRS) in China. METHODS: A total of 76 surgical tissue samples that included 43 CRSwNP samples (15 eosinophilic and 28 non-eosinophilic), 17 CRS samples without nasal polyps (CRSsNP), and 16 controls were obtained. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expression levels of Vγ1⁺γδT cells, Vγ4⁺γδT cells, eosinophil cationic protein (ECP), interleukin (IL)-8, transforming growth factor (TGF)-β2, metalloproteinase (MMP)-7, tissue inhibitor of metalloproteinase (TIMP)-4 and hypoxia-inducible factor (HIF)-1α. Enzyme linked immunosorbent assay (ELISA) was used to measure the protein level of ECP and MMP-7 in CRSwNP. The eosinophils were counted and the level of edema was analyzed with HE staining. RESULTS: The mRNA expression levels of the Vγ1 subset, ECP and MMP-7 were significantly increased in CRSwNP with histological characteristics of eosinophilic infiltration and edema. The expression of the Vγ1 gene in CRSwNP correlated positively with the expression of both ECP and MMP-7. No significant decreases in the mRNA expression levels of TGF-β2, TIMP-4 or HIF-1α were observed in the CRSwNP samples. The expression levels of Vγ1 gene, ECP and MMP-7 were significantly increased in eosinophilic CRSwNP compared to non-eosinophilic CRSwNP. CONCLUSIONS: Our results suggest the associations between Vγ1⁺γδT cells, ECP and MMP-7 in CRSwNP, indicating that Vγ1⁺γδT cells can induce the eosinophilic inflammation, which has a further effect on the formation of edema.
China ; Edema* ; Enzyme-Linked Immunosorbent Assay ; Eosinophil Cationic Protein* ; Eosinophils* ; Inflammation ; Interleukins ; Mucous Membrane ; Nasal Mucosa ; Nasal Polyps* ; RNA, Messenger ; Transforming Growth Factors

The contents of dihydromyricetin and total flavonoids of Hovenia acerba seeds were detected by HPLC-DAD and UV spectrophotometer. And then the correlation between the habitat, phenotype and quality of H. acerba seeds were deeply studied. There were big differences in both appearance and quality among the H. acerba seeds from different places. It showed that the content of dihydromyricetin in H. acerba seeds was 0.41-9.81 mg•g⁻¹, and the content of total flavonoids was 5.52-21.98 mg•g⁻¹. The cluster analysis showed that the quality of H. acerba seeds was related to the habitat. The samples from Jianghan Plain Area showed relatively stable and excellent quality. According to the correlation analysis, there was a significant positive correlation between 1 000-seeds weight, red-black seeds ratio and the content of dihydromyricetin, while the content of total flavonoids was not related to the 1 000-seeds weight and red-black seeds ratio. So the 1 000-seeds weight and red-black seeds ratio could be used as an initial judgment of the quality of H. acerba seeds. As dihydromyricetin and total flavonoids content was not related, both dihydromyricetin and total flavonoids should be taken into account when the quality of H. acerba seeds was studied.