[GAD67-GFP expression and co-localization with bNOS in main olfactory bulb of GAD67-GFP knock-in mouse].

OBJECTIVE: To investigate the distribution of GAD67 and the co-localization with bNOS in the main olfactory bulb of GAD67-GFP knock-in mouse. METHODS: Polymerase chain reaction was applied to identify the genotype of GAD67-GFP knock-in mouse, the animals were sacrificed and frozen sections of olfactory bulb were prepared. The Nissl-staining was performed to show an framework of the neuron in the olfactory bulb. The distribution of GAD67 and co-localization with bNOS were detected by immunofluorescence technique. RESULTS: The proportion of GAD67-positive cells among DAPI-positive cells were (42.98 ± 0.92)% in glomerular layer, (23.64 ± 0.84)% in mitral cell layer and (77.75 ± 0.84)% in granule cell layer; the bNOS-positive cells mainly existed in glomerular layer and mitral cell layer, very few in granule cell layer. No co-localization of GAD67 and bNOS in granule cell layer and mitral cell layer was found, but there was dispersed distribution in glomerular layer. CONCLUSION: GAD67-positive neurons mainly appear in glomerular layer and granule cell layer, and the bNOS is mostly expressed in glomerular layer and mitral cell layer; while the co-localization of GAD67 and bNOS only occurs in glomerular layer of olfactory bulb.

A Comparative Analysis of Two Machine Learning-Based Diagnostic Patterns with Thyroid Imaging Reporting and Data System for Thyroid Nodules: Diagnostic Performance and Unnecessary Biopsy Rate.

Background: The risk stratification system of the American College of Radiology Thyroid Imaging Reporting and Data System (ACR TI-RADS) for thyroid nodules is affected by low diagnostic specificity. Machine learning (ML) methods can optimize the diagnostic performance in medical image analysis. However, it is unknown which ML-based diagnostic pattern is more effective in improving diagnostic performance for thyroid nodules and reducing nodule biopsies. Therefore, we compared ML-assisted visual approaches and radiomics approaches with ACR TI-RADS in diagnostic performance and unnecessary fine-needle aspiration biopsy (FNAB) rate for thyroid nodules. Methods: This retrospective study evaluated a data set of ultrasound (US) and shear wave elastography (SWE) images in patients with biopsy-proven thyroid nodules (≥1 cm) from the Shanghai Tenth People's Hospital (743 nodules in 720 patients from September 2017 to January 2019) and an independent test data set from the Ma'anshan People's Hospital (106 nodules in 102 patients from February 2019 to April 2019). Six US features and five SWE parameters from the radiologists' interpretation were used for building the ML-assisted visual approaches. The radiomics features extracted from the US and SWE images were used with ML methods for developing the radiomics approaches. The diagnostic performance for differentiating thyroid nodules and the unnecessary FNAB rate of the ML-assisted visual approaches and the radiomics approaches were compared with ACR TI-RADS. Results: The ML-assisted US visual approach had the best diagnostic performance than the US radiomics approach and ACR TI-RADS (area under the curve [AUC]: 0.900 vs. 0.789 vs. 0.689 for the validation data set, 0.917 vs. 0.770 vs. 0.681 for the test data set). After adding SWE, the ML-assisted visual approach had a better diagnostic performance than US alone (AUC: 0.951 vs. 0.900 for the validation data set, 0.953 vs. 0.917 for the test data set). When applying the ML-assisted US+SWE visual approach, the unnecessary FNAB rate decreased from 30.0% to 4.5% in the validation data set and from 37.7% to 4.7% in the test data set in comparison to ACR TI-RADS. Conclusions: The ML-assisted dual modalities visual approach can assist radiologists to diagnose thyroid nodules more effectively and considerably reduce the unnecessary FNAB rate in the clinical management of thyroid nodules.

Highly efficient activation of peroxymonosulfate by Co3O4/Bi2WO6 p-n heterojunction composites for the degradation of ciprofloxacin under visible light irradiation.

Photocatalytic technology assisted via peroxymonosulfate (PMS) has good potential in water treatment. In this study, the Co3O4/Bi2WO6 composite was constructed via an in-situ calcination process and used to activate PMS for the degradation of ciprofloxacin (CIP) under visible light irradiation. The obtained 5 wt% Co3O4/Bi2WO6(CBWO-2) can highly effectively remove 86.2% CIP within 5 min visible light irradiation in presence of PMS. The excellent degradation performance of Co3O4/Bi2WO6/PMS system can be attributed to the synergistic effect between p-n heterojunction and PMS activation. The conduction band and valence band deviation between Co3O4 and Bi2WO6 were calculated by XPS techniques. Besides, DFT calculations were performed to further confirm the internal structure between Co3O4 and Bi2WO6. This work not only provides an approach to fabricate heterostructures but also indicated that Co3O4/Bi2WO6/PMS/Vis system is a potential environment remediation alternative for the efficient removal of recalcitrant organic compounds from wastewaters.

[Formaldehyde inhalation may damage olfactory bulb and hippocampus in rats].

OBJECTIVE: To investigate the effects of formaldehyde inhalation on the morphological damage, and Glu, GABA and NOS contents in olfactory bulb and hippocampus of rats. METHODS: Twenty SD rats were equally divided into two groups: rats in the control group inhaled fresh air, while the animals in experimental group were exposed to the air containing formaldehyde (12.5 mg/m(3), 4 h/d) for 7 days. Then rats were sacrificed and frozen sections of olfactory bulb and hippocampus were prepared. The morphological changes were examined and the Glu, GABA and NOS contents were detected using Nissl-staining, immunohistochemistry and Western blot, respectively. RESULT: Compared with the control group, there was a significant confusion and shrink of neuron morphology in experimental group, the number and staining intensity of Glu and NOS positive cells and protein contents were reduced. The protein expression of GABA was also decreased in the formaldehyde group. CONCLUSION: Formaldehyde inhalation can cause a severe morphological damage of olfactory bulb and hippocampus in SD rats,which may further impair memory and learning ability through the reduction of Glu, GABA and NOS expression.

Pregnancy outcomes of women with elevated second-trimester maternal serum alpha-fetoprotein.

OBJECTIVE: The aim of this study was to investigate the overall distribution of pregnancy outcomes in women with elevated second-trimester maternal serum alpha-fetoprotein (MS-AFP), and to determine the risk of adverse pregnancy outcomes (APOs) by MS-AFP level. MATERIALS AND METHODS: We retrospectively analyzed the clinical data of 429 women with elevated MS-AFP (≥2.5 multiple of the median (MOM)) and 1555 women with normal MS-AFP (0.5-2.49MOM) from a total of 46,741 prenatally screened singleton pregnant women. The overall distribution of APOs of the two groups, the risk of APOs by MS-AFP level, and the predictive value of elevated MS-AFP to APOs were analyzed. RESULTS: The incidence rate of APOs in elevated MS-AFP group was significantly higher than that in normal MS-AFP group (42.89 vs. 8.23%). In elevated MS-AFP group, the top three APOs, in term of incidence rate, were structural fetal abnormalities (7.93%), spontaneous abortion (7.46%) and preterm birth (7.23%); regarding to the risk, the top three APOs were stillbirth, spontaneous abortion and early-onset preeclampsia (odds ratio 35.98, 20.81 and 8.58 respectively). For structural fetal abnormalities, MS-AFP had predictive values for fetal open neural tube defects (ONTDs), gastroschisis and multiple malformations. CONCLUSION: Elevated MS-AFP is associated with increased risks of APOs. ONTDs complicate merely a small proportion of pregnancies with elevated MS-AFP, and the rest of them have high risks of obstetric complications. MS-AFP can help to identify these women at high risk of APOs in earlier second-trimester.

Histone deacetylase HDAC4 promotes the proliferation and invasion of glioma cells.

Glioma is the most lethal type of primary brain tumor characterized by aggressiveness and a poor prognosis. Histone deacetylase 4 (HDAC4) is frequently dysregulated in human malignancies. However, its biological functions in the development of glioma are not fully understood. The present study aimed to evaluate HDAC4 expression in human glioma and to elucidate the mechanistic role of HDAC4 in glioma. The results suggested that HDAC4 was significantly upregulated in glioma tissues and a number of glioma cell lines compared with adjacent non-tumor tissues and the non-cancerous human glial cell line SVG p12, respectively (P<0.05). The proliferation, adenosine triphosphate (ATP) levels and invasion ability were substantially enhanced in U251 cells with HDAC4 overexpression, and suppressed in U251 cells with a knockdown of HDAC4 compared with that in U251 cells transfected with the negative control. Knockdown of HDAC4 resulted in cell cycle arrest at the G0/G1 phase and induced the increase of reactive oxygen species level in U251 cells. Furthermore, HDAC4 overexpression was revealed to substantially inhibit the expression of cyclin-dependent kinase (CDK) inhibitors p21 and p27, and the expression of E-cadherin and ß­catenin in glioma U251 cells. Knockdown of HDAC4 substantially promoted the expression of CDK1 and CDK2 and vimentin in glioma U251 cells. Mechanistically, the results of the present study demonstrated that HDAC4 displayed a significant upregulation in glioma, and promoted glioma cell proliferation and invasion mediated through the repression of p21, p27, E-cadherin and ß­catenin, and the potentiation of CDK1, CDK2 and vimentin. Altogether, the present study revealed that HDAC4 overexpression was central for the tumorigenesis of glioma, which may serve as a useful prognostic biomarker and potential therapeutic target for glioma.

[Cultivation and karyotype analysis of the human embryonic stem cells HUES4].

OBJECTIVE: To establish a protocol for culturing of human embryonic stem cells (HUES4) without any animal-derived feeder cells and to investigate the karyotype stabilities of HUES4 cells after long-term cultivation. METHODS: HUES4 cells were cultured on mitomycin C treated MEFs or human foreskin fibroblast feeder cells. The pluripotency of the ES cells was analyzed by immunocytochemistry staining to detect the expression of pluripotent marker, karyotype of the ES cells at passage 27, 34, 41, 44 and short tandem repeat (STR) at passage 27 were analyzed. RESULTS: The HUES4 cells cultured on human feeder cells were positive for alkaline phosphatase activity, SSEA-4, TRA-1-60 and TRA-1-81 staining, but negative for SSEA-1. Analysis of karyotype at different passages suggested an abnormal karyotype 46, XY, t(9;15)(q22;q26) mosaicism occurred in HUES4, and the ratios of abnormal increased with passage. CONCLUSION: HUES4 could be cultured without animal-derived feeder cells and the incidence of abnormal karyotype might be increased with long-term culture.