Objective To observe the learning and memory ability of rats after injection of Aβ25-35 protein in different concentrations into the lateral ventricle assessed by Morris water maze test, and to explore the optimal concentration of Aβ25-35 in the preparation of AD model rats.Methods Male SD rats were randomly divided into sham operated group and model group.The rats of model group received Aβ25-35 injection in concentrations of 2 μg/μL, 4 μg/μL and 8 μg/μL, respectively.According to the Rat Brain Stereotaxic Atlas, 5 μL of aggregation of Aβ25-35 was injected into the right lateral ventricle to establish the AD rat model.7 days after successful modeling, Morris water maze was used to test thechanges of learning and memory ability of the rats.Results There was no significant difference in the average swimming speed between the two groups (P > 0.05).The escape latency time of rats in the model group was significantly increasedcompared with the sham group (P < 0.05).In the model group, the escape latency time of rats treated with 4 μg/μL and 8 μg/μL Aβ25-35 was significantly increased compared with the rats injected with 2 μg/μL (P < 0.05), while there was no significant difference between rats treated with 4 μg/μL and 8 μg/μL Aβ25-35 (P > 0.05).The activity time and distance of target quadrant of the rats injected with different concentration of Aβ25-35in the model group were significantly reduced compared with the sham group (P < 0.05), but no significant difference amongthe rats treated with different Aβ25-35 concentrations (P > 0.05).Compared with the sham-operated group, the number of platform-crossing of rats injected with different doses of Aβ25-35in the model group were significantly reduced (P < 0.05).In the model group, the rats treated with 4 μg/μL and 8 μg/μL was significantly reduced compared with the group with 2 μg/μL injection (P < 0.05).There was no significant difference between the rats injected with 4 μg/μL and 8 μg/μL (P > 0.05).Conclusions The recommended dose and concentration of Aβ25-35 to be injected into the unilateral ventricle to establisha rat model of Alzheimer's disease is 4 μg/μL in a volume of 5 μL.

Objective:To explore the expressions of Disabled-1 (Dab1 )in human breast epithelial cells and breast cancer cells,and to clarify its role in cell cycle.Methods:Real-time PCR was used to analyze the Dab1 mRNA expressions in breast epithelial cells MCF-10A and breast cancer cells MCF-7,BT-549,and MDA-MB-231. The Dab1 protein expressions in those cells were tested by Western blotting method. The BT-549 cells at logarithmic growth phase were divided into control,pKH3,and pKH3-Dab1 groups;the cell cycle was investigated by flow cytometry.Results:The Real-time PCR results showed that the Dab1 mRNA expression levels in MCF-7 cells (0.504 ± 0.037),BT-549 cells (0.302 ± 0.027),and MDA-MB-231 cells (0.330 ± 0.031 )were reduced compared with MCF-10A cells (0.998±0.020)(P <0.05).The Western blotting results showed that the Dab1 protein expression levels in breast cancer cells MCF-7 (0.134±0.014),BT-549 (0.076±0.01),and MDA-MB-231 (0.074±0.005)were reduced compared with MCF-10A cells (0.227±0.021)(P <0.05).Compared with control group and pKH3 group,the cell cycle in pKH3-Dab1 group was inhibited at G1 phase detected by flow cytometry analysis. Conclusion:The expression of Dab1 is down-regulated in breast cancer cells,and the over-expression of Dab1 can inhibit the cell cycle at G1 phase.

OBJECTIVETo investigate the effects of Fufang Banmao capusle on the proteome of SMMC-7721 cells and discover potential molecular mechanism of anti-cancer at molecular level.

METHODSMMC-7721 cells were treated by Fufang Banmao capusle serum prepared with serum pharmacological method; proteomic protocol involving 2-DE, image analysis and mass spectrometry were used to detect the proteins in cells influenced by Fufang Banmao capusle.

RESULTApproximately 450 protein spots in SMMC-7721 cells were resolved and detected in 2-D gel maps from pH3-10L IEF. 47 protein plots varied over 2-fold quantitively between treated sample and control sample were uncovered. 13 differentially expressed proteins spots were further identified by MALDI-TOF-MS analysis and four of them were successfully identified. Annexin A5, heatshock 70 x 10(3) protein 8 was significantly up-regulated in treated sample compared with control sample, while Eukaryotic translation initiation factor 5A and Peroxiredoxin-2 was significantly down-regulated in treated sample.

CONCLUSION4 differently expressed proteins associated with the proliferation, apoptosis, immunity of tumor were detected and they might provide clues for the coming research. The protocol of proteomics combined with serum pharmacological method is an effective platform to research complicated formulas in that it is capable of laying out many proteins associated with Fufang Banmao capusle.

Animals ; Annexin A5 ; metabolism ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Coleoptera ; chemistry ; Drug Combinations ; Electrophoresis, Gel, Two-Dimensional ; Female ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Peptide Initiation Factors ; metabolism ; Peroxiredoxins ; metabolism ; Proteome ; drug effects ; metabolism ; Proteomics ; methods ; RNA-Binding Proteins ; metabolism ; Rabbits ; Serum ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective:To compare the effect of chloroquine on apoptosis of normal gastric epithelial cells and gastric cancer cell line HGC-27. Methods:Change of these two kinds of cells were observed by inverted microscope after treating with CQ. HGC-27 cells were detected on the effect of apoptosis by DAPI nuclear staining after treating with CQ. The proliferation of cells were measured by CCK-8. Changes of mitochondrial membrane potential were investigated by JC-1 after treating with CQ. The expression of apoptosis protein effector enzyme Caspase-3 and substrate PARP in these two kinds of cells were tested by Western blot after using chloroquine (CQ) and rapamycin ( rapamycin, RAP ) to treat cells 72 h. Results: After treated with 10 μmol/L CQ 72 h, morphological characteristics of GES-1 cells and HGC-27 cells could be visible under the microscope,CQ induced apoptosis of GES-1 cells,on the contrary,it could make the HGC-27 cell get widened,the number of apoptotic cells gradually increased,the cell density decreased,cell atrophy and gradually turned round,cytoplasm reduced,at last,lose normal cell morphology. After two kinds of cells treated with CQ 72 h,as for GES-1 cell nuclei stained light,nuclear size and shape were not changed,however,HGC-27 nuclei showed pyknosis or granular fluorescence dense concentrated form. CCK-8 results showed that comparing with normal gastric epithelial cells GES-1,the pro-liferation of gastric cancer HGC-27 cells activity could be inhibited by CQ. JC-1 results showed that the change of the red fluorescence to green fluorescence in HGC-27 cells treated by CQ. Western blot showed that after being treated with CQ and RAP in normal gastric epithelial cells and HGC-27 cell line 72 h,the expression of apoptosis protein Caspase-3 and PARP in gastric cancer cell HGC-27 decreased significantly,comparing to that in GES-1 cells. Conclusion:Compared to normal gastric epithelial cells,CQ can inhibit human gastric cancer HGC-27 cell viability and induce apoptosis.

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Objective To examine the in vivo visual imaging of buccal carcinoma with the nearinfrared fluorescent quantum dots. Methods The U14 cells were labeled by endocytosis with QD800 (U14/QD800) which was linked with cell-penetrating peptide. Different number of U14/QD800 was injected under the buccal mucosa of nude mice and Kunming mice separately and imaged at different time to detect the in vivo sensitivity and dynamic imaging of U14/QD800. Results The minimum number of U14/QD800 cells which could be detected by in vivo imaging system was 1 × 104 in nude mice's cheek and 1 × 105 in Kunming mice's. The time for visual imaging of 1 × 104, 1 × 105 and 1 × 106 U14/QD800 cells in nude mice was 3, 7 and 16 d separately, and 3 and 10 d separately in Kunming mice. Conclusions Due to its strong tissue penetration, near-infrared fluorescent quantum dots have great prospects in cancer early diagnosis, visual observation and individual treatment.

Objective:To compare the diagnostic accuracy between multiparametric magnetic resonance imaging (mp-MRI) and biparametric magnetic resonance imaging (bp-MRI) in muscle-invasive bladder cancer (MIBC).Methods:The clinical data of 195 patients with bladder cancer at the First Affiliated Hospital of Nanjing Medical University from July 2020 to June 2022, were retrospectively reviewed. There were 160 males and 35 females, with the median age of 68(61, 76)years old. Mp-MRI was performed on each patient within 6 weeks before transurethral resection of bladder tumor or radical cystectomy. Each patients’ images were divided into two sets. Set 1 (bp-MRI) included the axial, sagittal, coronal T2-weighted images (T2WI), and axial diffusion-weighted images (DWI) or apparent diffusion coefficient maps. Set 2 (mp-MRI) included Set 1 images in addition to dynamic contrast-enhanced images. All images were independently reviewed and evaluated by two radiologists. Mp-MRI was evaluated according to the Vesical Imaging-Reporting and Data System (VI-RADS)guideline, and bp-MRI was evaluated according to two types of criteria. Bp-MRI (Criterion A): VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point. Bp-MRI (Criterion B): VI-RADS scoring is determined 3 when T2WI 3-point with DWI 2-point. VI-RADS scoring ≥ 3 or ≥ 4 was used as the cut-off value to predict MIBC. The sensitivity, specificity, positive predictive value, and negative predictive value of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were calculated, as well as receiver operating characteristic curves and the areas under the curve (AUC).Results:Of 195 patients, 135 patients (69.2%) were pathologically confirmed as NMIBC and 60 patients (30.8%) were MIBC. When the VI-RADS cut-off value was ≥ 3, the sensitivity of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were identical, all at 88.3% (53/60). The specificity of bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 88.9% (120/135), 73.3% (99/13), and 86.7% (117/135), respectively. When the VI-RADS cut-off value was ≥ 4, both bp-MRI (Criterion A) and bp-MRI (Criterion B) were classified as the same criterion. The sensitivity of bp-MRI and mp-MRI were 70.0% (42/60) and 75.0% (45/60), respectively. The specificity of bp-MRI and mp-MRI were identical, at 95.6% (129/135). The AUC for bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 0.927 (95% CI 0.881-0.959), 0.904 (95% CI 0.853-0.941), and 0.927 (95% CI 0.881-0.959), respectively. The AUC for bp-MRI (Criterion A) and mp-MRI were significantly higher than that of bp-MRI (Criterion B) ( P<0.001). There was no significant difference in AUC between bp-MRI (Criterion A) and mp-MRI ( P=0.939). Conclusions:Bp-MRI (Criterion A), VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point, shows comparable diagnostic accuracy in predicting MIBC with mp-MRI. Compared to bp-MRI (Criterion B), the corresponding situation when VI-RADS scoring is determined 3, bp-MRI (Criterion A) may have better diagnostic accuracy than bp-MRI (Criterion B) in predicting MIBC.

OBJECTIVETo investigate the main inhalant allergens and their distribution patterns in children with allergic diseases from Xi'an and the surrounding area and to provide evidence for the prevention and treatment of allergic diseases in children.

METHODSSkin prick test was performed using liquid with 13 standardized allergens (ALK-ABELL, Denmark) on 3085 children from Xi'an and the surrounding area who were treated for allergic diseases between July 2006 and July 2011, to detect inhalant allergens.

RESULTSOf the 3085 patients, 1368 (44.34%) had positive SPT results, with the most prevalent inhalant allergen being Dermatophagoides pteronyssinus (804 cases, 26.06%), followed by Dermatophagoides farinae (793 cases, 25.71%), Blomia tropicalis (440 cases, 14.26%), mugwort (282 cases, 9.14%), and cat hair (204 cases, 6.61%). The positive rates were 28.66% in the <4 years group, 41.85% in the 4-6 years group, and 58.61% in the 7-15 years group (P<0.01). Males had a significantly higher SPT positive rate than females (47.78% vs 38.50%；P<0.05). The SPT positive rate was highest in children with allergic rhinitis (72.41%), followed by bronchial asthma (62-25%), allergic dermatosis (45.83%), and allergic purpura (36.28%).

CONCLUSIONSIn children from Xi'an and the surrounding area, the main inhalant allergens for allergic diseases include Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, mugwort and cat hair. The SPT positive rate increases with age. Male children have a higher SPT positive rate than female children. The SPT positive rate is highest in children with allergic rhinitis.

Adolescent ; Allergens ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Hypersensitivity ; diagnosis ; Infant ; Male ; Skin Tests

Objective:To explore the empathy of patients with cerebral small vessel disease (CSVD) and its relationship with cognitive functions.Methods:A total of 35 patients with CSVD and 26 normal controls with matching gender, age and education were enrolled.The Chinese version of the interpersonal reactivity index (IRI-C) and the multifaceted empathy test(MET) were used to assess the empathy of the participants.The montreal cognitive assessment scale (MoCA), the Hamilton anxiety scale(HAMA), and the Hamilton depression scale (HAMD) were applied to assess participants' overall cognitive function and emotional state.SPSS 19.0 software was used to analyze the differences between the CSVD group and the control group, while the influencing factors of empathy were studied by Pearson correlation analysis.Results:The total scores of IRI, perspective taking(PT), fantasy(FS) and empathy concern(EC) of CSVD patients ((37.25±11.71), (6.94±4.35), (9.45±4.68) and (16.40±4.34)) were lower than those of the control group ((50.61±11.07), (11.84±3.90), (13.23±5.01), (19.69±3.03)), and the differences were statistically significant (IRI score: t=4.506, P<0.05.PT: t=4.539, P<0.05.FS: t=3.021, P<0.05.EC: t=3.308, P<0.05). The personal distress(PD) scores of CSVD group were (4.68±4.16), while(6.00±4.69) in control group, and the difference was not significant ( t=1.154, P>0.05). The MET-C results showed that there was a difference in the correct number of empathic emotion recognition between the two groups(CVSD group: (25.08±6.77), control group: (32.30±3.42), t=4.978, P<0.05), however, there was no significant difference between the two groups in emotional empathy scores ( t=1.390, P>0.05). Pearson correlation analysis showed that the total score of IRI and PT in the CSVD group were positively correlated with education level ( r=0.374, 0.471, both P<0.05). PT was positively correlated with MoCA score ( r=0.458, P=0.006). PD was positively correlated with HAMA score and HAMD score ( r=0.521, 0.541, both P<0.05). The correct number of emotion recognition was positively correlated with education level ( r=0.600, P<0.001) and MoCA score ( r=0.665, P<0.001), and negatively correlated with HAMA score( r=-0.445, P<0.05) and HAMD score ( r=-0.421, P<0.05). Conclusion:The empathy of patients with CSVD is lower than that of the normal group, and it is manifested as a decline in cognitive empathy, which is positively related to the overall cognitive function.