OBJECTIVE: Brain-derived neurotrophic factor (BDNF), one of the most abundant and important neurotrophins, is known to be involved in the development, survival, maintenance, and plasticity of neurons in the nervous system. Some studies have suggested that BDNF may play a role in the pathophysiology of several psychiatric illnesses such as depression and schizophrenia. Similarly, it is likely that the alteration of BDNF may be associated with the neuro-modulation that contributes to the development of somatization disorder. METHODS: The purpose of this study was to determine whether there is an abnormality of plasma BDNF levels in patients with somatization disorder, and to analyze the nature of the alteration after pharmacotherapy using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The plasma BDNF levels of the patients with a somatization disorder were significantly lower compared with those of the control volunteers (83.61±89.97 pg/mL vs. 771.36±562.14 pg/mL); moreover, the plasma BDNF levels of those patients who received an antidepressant were significantly increased after the treatment (118.13±91.45 pg/mL vs. 72.92±88.21 pg/mL). CONCLUSION: These results suggest that BDNF may play a role in the pathophysiology of somatization disorder.
Brain-Derived Neurotrophic Factor* ; Depression ; Drug Therapy ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nerve Growth Factors ; Nervous System ; Neurons ; Plasma* ; Plastics ; Schizophrenia ; Somatoform Disorders* ; Volunteers

Purpose: Deodeok (Codonopsis lanceolata) is generally used in conventional medicines and is considered to have remedial properties to cure several diseases. However, application of the C. lanceolata bud as a novel food ingredient has not been fully explored. Hydrogen peroxide (H2O2) is associated with the production of oxidative damage that results in mutagenesis, carcinogenesis, and cell death. This study examines the neuroprotective effect of C. lanceolata bud extracts (CLBE) on H2O2 -stimulated apoptosis in SH-SY5Y cells. Methods: C. lanceolata bud of length 10 to 15 cm was collected and extracted using 70% ethanol. Cytotoxicity was evaluated by the EZ-cytox reagent, measurement of lactic dehydrogenase (LDH) release and reactive oxygen species (ROS). The morphological changes of the nuclei were determined using the Hoechst 33258 dye. Enzyme activities were analyzed using the caspase activity assay kit. Related protein expressions were quantified by the Western blot immunoassay in H2O2 -stimulated SH-SY5Y cells. Results: Cell viability, LDH release and ROS generation, demonstrated neuroprotective effects of CLBE in H2O2-stimulated SH-SY5Y cells. The occurrence of apoptosis in H2O2-stimulated cells was confirmed by caspase activity, which was increased in H2O2-stimulated SH-SY5Y cells compared to the unexposed group. Pretreatment of CLBE was observed to inhibit the H2O2–stimulated apoptosis. In addition, exposure to CLBE resulted in increased expression of the Bcl-2 (B cell lymphoma 2) protein and decreased expression of the Bax (Bcl2 associated X) protein. Conclusion This study shows that exposure to CLBE alleviates the H2O2-stimulated neuronal damage in SH-SY5Y cells. Our results indicate the potential application of CLBE in neurodegenerative disease therapy or prevention.

Purpose: The ginger rhizome (Zingiber officinale) is widely cultivated as a spice for its aromatic and pungent components. One of its constituents, 6-hydroxydopamine (6-OHDA) is usually thought to cross the cell membrane through dopamine uptake transporters, and induce inhibition of mitochondrial respiration and the generation of intracellular reactive oxygen species (ROS). This study examines the neuroprotective effect and acetylcholinesterase (AChE) inhibitory activity of fermented ginger extracts (FGEs) on 6-OHDA induced toxicity in SH-SY5Y human neuroblastoma cells. Methods: Ginger was fermented using 2 species of Bacillus subtilis, with or without enzyme pretreatment. Each sample was extracted with 70% ethanol. Neurotoxicity was assessed by applying the EZ-Cytox cell viability assay and by measuring lactic dehydrogenase (LDH) release. Morphological changes of apoptotic cell nuclei were observed by Hoechst staining. Cell growth and apoptosis of SH-SY5Y cells were determined by Western blotting and enzyme activity analysis of caspase-3, and AChE enzymatic activity was determined by the assay. Results: In terms of cell viability and LDH release, exposure to FGE showed neuroprotective activities against 6-OHDA stimulated stress in SH-SY5Y cells. Furthermore, FGE reduced the 6-OHDA-induced apoptosis, as determined by Hoechst staining. The occurrence of apoptosis in 6-OHDA treated cells was confirmed by determining the caspase-3 activity. Exposure to 6-OHDA resulted in increased caspase-3 activity of SH-SY5Y cells, as compared to the unexposed group. However, pre-treatment with FGE inhibited the activity of caspase-3. The neuroprotective effects of FGE were also found to be caspase-dependent, based on reduction of caspase-3 activity. Exposure to FGE also inhibited the activity of AChE induced by 6-OHDA, in a dose-dependent manner. Conclusion Taken together, our results show that FGE exhibits a neuroprotective effect in 6-OHDA treated SH-SY5Y cells, thereby making it a potential novel agent for the prevention or treatment of neurodegenerative disease.

Objective:To understand the genotyping and molecular epidemiological characteristics of Norovirus (NoV) GⅡ in adult acute gastroenteritis outpatients in Shanghai from 2015 to 2016.Methods:A total of 912 stool specimens from adult patients with acute diarrhea from March 2015 to December 2016 (431 in 2015 and 481 in 2016) were collected. The Norovirus GⅡ type was detected by one-step quantitative reverse transcription PCR assay and the RdRp region of open reading frame (ORF) 1 and 5′end of ORF2 were amplified. The region was sequenced and classified.Results:From March 2015 to December 2016, NoV GⅡ were detected in 17.76% (162/912) of the samples, 15.08% (65 /431) in 2015 and 20.17% (97/481) in 2016. Based on sequence analysis of the RdRp and capsid sequences, 145 identified NoV strains were divided into 10 genotypes: GⅡ.Pe_GⅡ.4 Sydney 2012 (60), GⅡ.P17_GⅡ.17 (45), GⅡ.P16_GⅡ.13 (10), GⅡ.P12_GⅡ.3 (10), GⅡ.P7_GⅡ.6 (9), GⅡ.P21_GⅡ.21 (5), GⅡ.P16_GⅡ.4 Sydney 2012 (2), GⅡ.Pe_GⅡ.17 (2), GⅡ.P2_GⅡ.2 (1), and GⅡ.P7_GⅡ.14 (1).Conclusions:The main epidemic NoV GⅡ genotype in Shanghai was GⅡ.P17_GⅡ.17 in the spring of 2015. GⅡ.Pe_GⅡ.4 Sydney 2012 and GⅡ.P17_GⅡ.17 were identified as predominant genotypes during the winter of 2015 and spring of 2016. The most common genotype was GⅡ.Pe_GⅡ.4 Sydney 2012 in autumn of 2016. Continuous NoV outbreak surveillance is important for identifying changing trends in genotype distribution and emerging new strains.

Objective:To construct mitochondrial antiviral signaling ( MAVS) gene or Nuclear factor kappa B1 ( NFκB1) gene knockout Madin-Darby canine kidney (MDCK) cells, and identify the cell growth and proliferation characteristics of influenza virus (Flu) in these cells. Methods:MAVS or NFκB1 knockout MDCK cells were established using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) technique. Flu was inoculated into these cells, then the hemagglutination (HA) titers and TCID 50 were determined and compared with the original MDCK cells. Results:MDCK cells knocked out of MAVS (MDCK- MAVS-) or NFκB1 (MDCK- NFκB1 -) were obtained stably. The two cells were both adherent epithelioid cells as well as wild type MDCK cells. The HA titers showed no obvious difference among these cells after inoculating Flu. Nevertheless, the TCID 50 titers were respectively increased 9.5 times in MDCK- MAVS-cell culture and 10 times in MDCK- NFκB1 - cell culture, compared to the wild type MDCK cells. Conclusions:The infectivities of Flu were both increased in MDCK- MAVS-cells and MDCK- NFκB1 - cells, illustrating their potential in Flu culture.

BACKGROUND: Intestinal organoids have evolved as potential molecular tools that could be used to study host-microbiome interactions, nutrient uptake, and drug screening. Gut epithelial barrier functions play a crucial role in health and diseases, especially in autoimmune diseases, such as inflammatory bowel diseases (IBDs), because they disrupt the epithelial mucosa and impair barrier function. METHODS: In this study, we generated an in vitro IBD model based on dextran sodium sulfate (DSS) and intestinal organoids that could potentially be used to assess barrier integrity. Intestinal organoids were long-term cultivated and characterized with several specific markers, and the key functionality of paracellular permeability was determined using FITC-dextran 4 kDa. Intestinal organoids that had been treated with 2 lM DSS for 3 h were developed and the intestinal epithelial barrier function was sequentially evaluated. RESULTS: The results indicated that the paracellular permeability represented epithelial characteristics and their barrier function had declined when they were exposed to FITC-dextran 4 kDa after DSS treatment. In addition, we analyzed the endogenous mRNA expression of pro-inflammatory cytokines and their downstream effector genes. The results demonstrated that the inflammatory cytokines genes significantly increased in inflamed organoids compared to the control, leading to epithelial barrier damage and dysfunction. CONCLUSION The collective results showed that in vitro 3D organoids mimic in vivo tissue topology and functionality with minor limitations, and hence are helpful for testing disease models.

BACKGROUND: Intestinal organoids have evolved as potential molecular tools that could be used to study host-microbiome interactions, nutrient uptake, and drug screening. Gut epithelial barrier functions play a crucial role in health and diseases, especially in autoimmune diseases, such as inflammatory bowel diseases (IBDs), because they disrupt the epithelial mucosa and impair barrier function. METHODS: In this study, we generated an in vitro IBD model based on dextran sodium sulfate (DSS) and intestinal organoids that could potentially be used to assess barrier integrity. Intestinal organoids were long-term cultivated and characterized with several specific markers, and the key functionality of paracellular permeability was determined using FITC-dextran 4 kDa. Intestinal organoids that had been treated with 2 lM DSS for 3 h were developed and the intestinal epithelial barrier function was sequentially evaluated. RESULTS: The results indicated that the paracellular permeability represented epithelial characteristics and their barrier function had declined when they were exposed to FITC-dextran 4 kDa after DSS treatment. In addition, we analyzed the endogenous mRNA expression of pro-inflammatory cytokines and their downstream effector genes. The results demonstrated that the inflammatory cytokines genes significantly increased in inflamed organoids compared to the control, leading to epithelial barrier damage and dysfunction. CONCLUSION The collective results showed that in vitro 3D organoids mimic in vivo tissue topology and functionality with minor limitations, and hence are helpful for testing disease models.

Objective To investigate the difference in visual quality between oval and round corneal flaps in femtosecond excimer laser in situ keratomileusis(FS-LASIK).Methods In a prospective randomized controlled clinical study,patients who voluntarily underwent FS-LASIK surgery in Wuhan University Affiliated Aier Ophthalmology Hospital from June 2022 to February 2023 were randomly divided into oval valve group and round valve group,with 36 patients(72 eyes)in each group.During FS-LASIK,oval and round corneal flaps were made,and excimer laser surgery was performed in parallel.Observe intraoperative and postoperative complications.The unaided visual acuity(UCVA)and spherical equivalent(SE)at 1 day,1 week,1 month and 3 months after surgery were compared between the two groups at 3 months after surgery,and the modulation transfer function(MTF),Strel ratio(SR)and total higher-order aberration at 3 months after surgery were compared between the two groups.Results All patients had successfully completed the surgery without serious complications.There was no significant difference between the oval valve group and the round valve group in UCVA at 1 day,1 week after surgery,1 month after surgery,3 months after surgery.There was no significant difference between the two groups on the postoperative SE at 1 day,1 week after surgery,1 month after surgery,and 3 months after surgery.There was no significant difference in MTF,SR and total higher-order aberration between the two groups at 3 months after surgery.Conclusion Ideal visual acuity can be achieved with either oval or round corneal flap during FS-LASIK,and there is no significant difference in visual quality between the two in the early postoperativeperiod.

BACKGROUND: Papillary thyroid carcinoma (PTC) of the thyroid is the most common endocrine malignancy. High prevalence of an activating point mutation of BRAF gene, BRAFV600E, has been reported in PTC. We assessed the efficiency of peptide nucleic acid clamp real-time polymerase chain reaction (PNAcqPCR) for the detection of BRAFV600E mutation in PTC in comparison with direct sequencing (DS). METHODS: A total of 265 thyroid lesions including 200 PTCs, 5 follicular carcinomas, 60 benign lesions and 10 normal thyroid tissues were tested for BRAFV600E mutation by PNAcqPCR and DS. RESULTS: The sensitivity and accuracy of the PNAcqPCR method were both higher than those of DS for the detection of the BRAFV600E mutation. In clinical samples, 89% of PTCs harbored the BRAFV600E mutation, whereas 5 follicular carcinomas, 50 benign lesions and 10 normal thyroid tissues lacked the mutation. The mutation was associated with aggressive clinical behaviors as extrathyroid invasion (p=0.015), lymph node metastasis (p=0.002) and multiple tumor numbers (p=0.016) with statistical significance. CONCLUSIONS: The PNAcqPCR method is efficiently applicable for the detection of the BRAFV600E mutation in PTCs in a clinical setting.
Carcinoma ; Factor IX ; Lymph Nodes ; Neoplasm Metastasis ; Peptide Nucleic Acids ; Point Mutation ; Prevalence ; Real-Time Polymerase Chain Reaction ; Thyroid Gland ; Thyroid Neoplasms