Objective To evaluate the clinical diagnostic role of dental computed tomography (CT) and three-dimensional reconstruction (3DR) compared with conventional dental periapical radiography (DPR) in the diagnosis of molar furcation involvement (FI). Methods Dental CT and 3DR were performed about a maxillary first molar and its alveolar bone diagnosed as FI II? by conventional probe technique, and the images were compared with that of DPR. Results The images of dental CT scans and 3DR showed not only the area, type and quantity of periodontal destruction, but also the complex stereoscopic anatomical vision and ambient spatial relationship around the molar. Especially, the images displayed the alveolar destructions of the dental buccal and palate site that could not always be demonstrated by DPR. The degree of the molar FI was defined as III? in fact. Conclusion Dental CT and 3DR have many superiorities and value, which can not be replaced by DPR, in the clinical diagnosis and prognostic judgment of molar furcation involvement.

We studied the actions of diffusion weighted imaging (DWI) and dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI) in differentiating breast tumors. From January 2010 to February 2012, we retrospectively analyzed data of 95 cases with breast tumor pathologically confirmed from DWI and DCE-MRI. We compared the ADC value, time-intensity curve (TIC) and DCE-MRI parameters between breast tumors, and calculated the sensitivity and specificity for differentiating breast tumors. The results were as follows: (1) On DWI, mean ADC value of malignant tumor was lower than that of benign tumor (P < 0.05). For differentiating breast malignant tumors from benign neoplasm, a cut-off ADC value of 1.2 x 10(-3) mm2/s achieved a sensitivity of 74.1% and specificity of 70.3%. (2) On DCE-MRI, early enhancement ratio (EER) value of malignant tumor was higher than that of benign tumor whereas value of time to peak (Tpeak) and maximal enhancement ratio (SImax) were lower than that of benign tumor (all P < 0.05). As for TIC, type II and III were more frequently seen in malignant tumor than in benign tumor whereas type I was more common in benign tumor than in malignant tumor (all P < 0.05). For differentiating breast malignant tumors from benign neoplasm, DCE-MRI obtained a sensitivity of 89.7% and specificity of 70.3%. (3) For differentiating breast malignant tumors from benign neoplasm, ADC value together with TIC obtained a sensitivity of 79.3% and specificity of 78.4%. Malignant or benign breast tumors could have their own unique characteristics on DWI and DCE-MRI. These characteristics might be helpful for differentiating these tumors.
Breast Neoplasms ; classification ; diagnosis ; Diagnosis, Differential ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Retrospective Studies ; Sensitivity and Specificity

This paper aims to investigate the value of diffusiion weighted imaging (DWI) and different apparent diffusion coefficient (ADC) methods to predict the curative effects of neoadjuvant chempotherapy (NAC) for breast cancer. From March 2010 to December 2012, seventy-one patients were pathologically confirmed invasive breast cancer by needle puncture biopsy received before surgery, and underwent magnetic resonance before and after NAC, the ADC were measured by mean ADC method and lower ADC method. The pathologic response after NAC was divided to major histological response (MHR) group and non-major histological response (NMHR) group according to Miller & Payne system. Results displayed that ADC values obtained before NAC, at the end of the second cycle of NAC, and after whole course of treatment, had good correlations between mean and lower ADC methods (the Pearson's correlation=0.699, 0.749 and 0.895, respectively). Significant difference in ADC obtained both with mean and lower ADC methods could be found between MHR and NMHR groups after the second cycle of NAC (P< 0.05). After the second cycle of NAC, significant difference in the change rate of ADC could be found between MHR and NMHR groups by using lower ADC method (P<0.05), but not be found by using mean ADC method (P >0.05). In conclusion, DWI could monitor the pathologic changes of breast cancer after NAC, and the lower ADC method might be used to evaluate the curative effect of NAC with the change rate of ADC.
Breast Neoplasms ; drug therapy ; pathology ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Neoadjuvant Therapy

Objective To construct dual-luciferase reporter plasmids containing the wild type and mutant rat extracellular signal-regulat-ed kinase 1 (ERK1) gene 3' untranslated regions (UTR) which were used to detect rno-miR-15b-5p's putative target gene. Methods The rat ERK1 gene 3' UTR fragment was amplified by polymerase chain reaction (PCR) from PC12 cell cDNA and cloned into pmiR-RB-ReportTM vector. The mutant rat ERK1 gene 3' UTR fragment was obtained by overlap PCR and inserted into pmiR-RB-ReportTM vector. Successful wild type and mutant recombinant plasmids were confirmed by DNA sequencing. PC12 cells were co-transfected with rno-miR-15b-5p mim-ic and pmiR-ERK13' UTR or pmiR-ERK1-mut 3' UTR and then analyzed by dual-luciferase reporter assay system. The achieved mutation sequence of the target site TGCTGCT was mutated to CGAACGT and GTACACG, respectively. Results The wild-type reporter vector pmiR-ERK13' UTR and the mutant reporter vector pmiR-ERK1-mut 3' UTR were successfully constructed. The rno-miR-15b-5p mimic de-creased the activity of pmiR-ERK13' UTR plasmid (P<0.001) but did not decrease the activity of pmiR-ERK1-mut 3' UTR plasmid. Conclu-sion The recombinant pmiR-ERK13' UTR and pmiR-ERK1-mut 3' UTR plasmids were constructed successfully, and luciferase activities demonstrated that the 3' UTR of ERK1 gene might be a potential target of rno-miR-15b-5p.

Objective:To investigate the stability of α-synuclein (α-Syn) aggregates formed by incubation with blood plasma of patients with Parkinson's disease (PD).Methods:Peripheral blood samples were collected from 10 patients diagnosed as having PD in our hospital from June 2020 to December 2020 and 10 healthy control subjects (HC) at the same time period. The 1 mg recombinant human α-Syn was dissolved in 140 μL 0.01 mol/L phosphate buffer solution (PBS), and then, incubated with plasma from HC and PD patients and PBS at 37 ℃ for 7 d (HC group, PD group and PBS group). Preformed fiber (PFF) group was used for subsequent experiment with 10 μg PFF. After digestion with different concentrations of trypsin (concentration ratios of trypsin/α-Syn=1:80, 1:40, and 1:20) and protease K (PK, 1.0, 1.5, and 2.0 μg/mL), the differences of α-Syn levels before and after digestion were detected by Western blotting.Results:(1) Effect of trypsin on PFF digestion: PFF gradually decreased with the increase of trypsin doses; when trypsin/α-Syn ratio=1:20, α-Syn aggregates with molecular weight greater than 35 000 were almost completely digested, and its digestion was significantly different as compared with that in other concentration ratios ( P<0.05). (2) Effect of trypsin on digestion of α-Syn aggregates: at the relative molecular weight<25 000, when the concentration ratio of trypsin/α-Syn=1:20, as compared with HC and PD groups, the PBS group had significantly more obvious decrease ( P<0.05). At the relative molecular weight 35 000-40 000, the α-Syn levels in PBS, HC and PD groups were significantly decreased as compared with those before digestion at all concentration ratios; as compared with HC and PD groups, the PBS group had significantly more obvious decrease ( P<0.05). At molecular weight>40 000, α-Syn decreased significantly in PD group only when the concentration ratio of trypsin/α-Syn=1:20, and the degrees of digestion was PBS group>HC group>PD group, with significant differences among groups ( P<0.05). (3) Effect of PK on PFF digestion: when PK concentration was 1.0, 1.5 or 2.0 μg/mL, α-Syn was basically digested at relative molecular weight>35 000, and α-Syn monomer was reduced and small fragment appeared at relative molecular weight<25 000; as compared with negative controls (0 μg/mL PK), these changes in groups of PK concentration of 1.0, 1.5 or 2.0 μg/mL were significantly different ( P<0.05). (4) Effect of PK on digestion of α-Syn aggregates: at relative molecular weight<25 000, α-Syn in PBS group was digested into smaller fragments while α-Syn in HC and PD groups was not digested; significant differences was noted among groups at the same concentration ( P<0.05). At relative molecular weight of 35 000-40 000, with the increase of PK concentration, the amount of α-Syn dimer in PBS group decreased (increased digestibility), that in HC group increased, however, that in PD group did not change obviously; significant difference was noted at the same concentration among the three groups ( P<0.05). At relative molecular weight>40 000, with the increase of PK concentration, α-Syn in PBS, HC and PD groups decreased to a certain extent, and significant difference was noted among groups at the same concentration ( P<0.05). Conclusion:The stability of α-Syn aggregates formed by incubation with plasma from PD patients is higher than that formed by incubation with HC plasma and PBS.

Objective:To explore the mechanism of miR-15b-5p involving in depression-like behavior in Parkinson's disease (PD).Methods:(1) Eighteen C57BL/6N mice were randomly divided into PD group, intervention group and control group ( n=6). PD models in PD group were established by stereotaxically injecting 0.25 mg/kg rotenone into the right striatum; mice in the intervention group were injected with 0.25 mg/kg rotenone and miR-15b-5p inhibitor lentivirus, while mice in the control group were injected with equal volume of PBS. Four weeks after that, open field test and rotarod test were performed to evaluate the motor ability, and sucrose preference and forced swimming tests were performed to evaluate the depression-like behaviors. And then, proteins and miRNAs in the substantia nigra were extracted; real-time fluorescent quantitative reverse transcription PCR (qRT-PCR) was used to detect the miR-15b-5p expression,and Western blotting and immunofluorescent staining were used to detect the tyrosine hydroxylase (TH), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and postsynaptic density protein 95 (PSD95) protein expressions. (2) The 100 nmol/L miR-15b-5p mimic/inhibitor and their negative control sequences were transfected into SH-SY5Y cells on 6-well plates (named miR-15b-5p mimic group, miR-15b-5p mimic control group, miR-15b-5p inhibitor group and miR-15b-5p inhibitor control group, respectively); 48 h after that, BDNF, TrkB and PSD95 protein expressions were detected by Western blotting and miR-15b-5p expression by qRT-PCR. (3) The 100 ng BDNF 3'-UTR wild-type or mutant luciferase reporter vector plasmids and 100 nmol/L miR-15b-5p mimic/inhibitor or their negative control sequences were co-transfected into SH-SY5Y cells on 24-well plates, and luciferase reporter activity assay was performed 48 h after co-transfection to detect the luciferase activity. Results:(1) Compared with the control group, the PD group had significantly reduced movement speed, shortened rotarod drop latency, decreased percentage of sucrose preference, and prolonged immobility time ( P<0.05); compared with the PD group, the intervention group had significantly increased movement speed, prolonged rotarod drop latency, increased percentage of sucrose preference, and shortened immobility time ( P<0.05). (2) Compared with the miR-15b-5p mimic control group, the miR-15b-5p mimic group had significantly increased miR-15b-5p expression, and decreased BDNF, TrkB and PSD95 protein expressions (100.00±5.75 vs. 66.79±5.90; 100.00±5.95 vs. 84.46±5.77; 100.00±7.02 vs. 80.43±3.25, P<0.05). Compared with the miR-15b-5p inhibitor control group, the miR-15b-5p inhibitor group had significantly decreased miR-15b-5p expression, and increased BDNF, TrkB and PSD95 expressions (100.00±6.81 vs. 119.90±5.66; 100.00±2.88 vs. 110.10±4.15; 100.00±2.19 vs. 124.60±11.69, P<0.05). Compared with the control group, PD group had significantly increased miR-15b-5p expression, and significantly decreased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (100.00±9.20 vs. 63.60±12.80; 100.00±9.88 vs. 71.95±10.00; 100.00±5.16 vs. 70.37±8.43; 100.00±7.01 vs. 68.12±10.22; 100.00±12.99 vs. 48.23±12.58) in the substantia nigra ( P<0.05); compared with the PD group, the intervention group had significantly lower miR-15b-5p expression and increased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (63.60±12.80 vs. 90.69±9.84; 71.95±10.00 vs. 93.31±4.50; 70.37±8.43 vs. 88.11±4.10; 68.12±10.22 vs. 89.59±5.93; 48.23±12.58 vs. 83.65±10.52) in the substantia nigra ( P<0.05). (3) Compared with the BDNF 3'-UTR wild-type+miR-15b-5p mimic control group, the BDNF 3'-UTR wild-type+miR-15b-5p mimic group had significantly decreased luciferase activity (100.00±5.07 vs. 90.59±1.75, P<0.05); compared with the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor control group, the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor group had significantly increased luciferase activity (100.00±5.08 vs. 152.20±31.87, P<0.05). Conclusion:MiR-15b-5p is involved in depression-like behavior in PD by down-regulating the BDNF/TrkB/PSD95 expressions.

Objective:To explore the role of α-synuclein (α-Syn) in pathogenesis of Parkinson's disease (PD) and multiple system atrophy (MSA) by observing the ability of α-Syn aggregates incubated with PD and MSA patients' plasma to destroy cell membrane.Methods:Peripheral blood samples were collected from 5 PD patients and 5 MSA patients diagnosed in Department of Neurology, Affiliated Hospital of Guilin Medical University from January 2018 to January 2022, as well as 5 physical examination healthy control subjects (HCs) during the same period. The α-Syn was dissolved in 0.01 mol/L PBS and then incubated with PBS, and plasma from HCs, PD patients and MSA patients at 37 ℃ for 4 d, respectively (named as PBS group, HC group, PD group and MSA group). Small unilamellar vesicles (SUVs) containing calcein were prepared with acidic phospholipid, 1-palmityl-2-oleoyl-Sn-glycerol-3-phospho-L-serine (POPS), by membrane dispersion-ultrasonic method; the particle size and morphology of SUVs were analyzed by Malvern Zetasizer Nano ZS and transmission electron microscope. SUVs and human neuroblastoma cells (SH-SY5Y) were treated with different concentrations of α-Syn aggregates (0.5, 1.0, 2.0, 4.0, 8.0 and 16.0 μmol/L). The abilities of α-Syn aggregates formed under different conditions to destroy liposomes and cell membrane were evaluated by measuring the calcein relative release and intracellular calcein fluorescent value after dialysis.Results:(1) Destructive effect of α-Syn aggregates on SUVs: calcein induced by α-Syn aggregates in each group increasingly released with the increase of protein concentration; at protein concentration of 8 μmol/L, the calcein released from SUVs in PD and MSA groups was significantly higher than that in PBS and HC groups, and the calcein released from SUVs in MSA group was further significantly higher than that in the PD group ( P<0.05). (2) Destructive effect of α-Syn aggregates on cell membrane in SH-SY5Y: the calcein fluorescent value in each group decreased with the increase of protein concentration; at protein concentration of 8 μmol/L, the intracellular calcein fluorescent value in PD and MSA groups were significantly decreased compared with that in PBS and HC groups ( P<0.05); the intracellular calcein fluorescent value in MSA group was further significantly decreased compared with that in PD group ( P<0.05). (3) Effect of α-Syn aggregates on SH-SY5Y cell survival: at protein concentration of 8 μmol/L, the viability of SH-SY5Y cells in each group decreased obviously; PD and MSA groups had significantly decreased cell viability compared with PBS and HC groups ( P<0.05); and the viability in MSA group was further statistically decreased compared with that in PD group ( P<0.05). Conclusion:The ability of α-Syn aggregates incubated with PD and MSA patients' plasma to destroy cell membrane is greater than that with HCs' plasma, especially those with MSA patients' plasma.

ETS-1 is a transcription factor that is a member of the E26 transformation-specific (ETS) family. Galanin receptor 2 (GalR2), a subtype of receptors of the neuropeptide galanin, has been shown to have an antidepressant-like effect after activation in rodents. Our previous study has shown that overexpression of ETS-1 increases the expression of GalR2 in PC12 phaeochromocytoma cells. However, whether ETS-1 has an antidepressant-like effect is still unclear. In this study, we found that chronic mild stress (CMS) decreased the expression of both ETS-1 and GalR2 in the ventral hippocampus of rats. Meanwhile, we demonstrated that overexpression of ETS-1 increased the expression of GalR2 in primary hippocampal neurons. Importantly, we showed that overexpression of ETS-1 in the ventral hippocampus counteracted the depression-like behaviors of CMS rats. Furthermore, we found that overexpression of ETS-1 increased the level of downstream phosphorylated extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2) of GalR2 in the ventral hippocampus of CMS rats. Taken together, our findings suggest that ETS-1 has an antidepressant-like effect in rats, which might be mediated by increasing the level of GalR2 and its downstream p-ERK1/2 in the ventral hippocampus.

We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpi, tcdA, tcdB, cdtA, cdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C. difficile infection and 45 isolates. The results were compared with a reference standard combined with BD MAX Cdiff, real-time cell analysis assay (RTCA), and mPCR-CE. The toxigenic C. difficile species were detected in 36 isolates and 45 stool specimens by the mPCR-CE, which provided a positive rate of 20.3% (81/399). The mPCR-CE had a specificity of 97.2% and a sensitivity of 96.0%, which was higher than RTCA (x = 5.67, P = 0.017) but lower than BD MAX Cdiff (P = 0.245). Among the 45 strains, 44 (97.8%) were determined as nonribotype 027 by the mPCR-CE, which was fully agreed with PCR ribotyping. Even though ribotypes 017 (n = 8, 17.8%), 001 (n = 6, 13.3%), and 012 (n = 7, 15.6%) were predominant in this region, ribotype 027 was an important genotype monitored routinely. The mPCR-CE provided an alternative diagnosis tool for the simultaneous detection of toxigenic C. difficile in stool and potentially differentiated between RT027 and non-RT027.
Clostridium Infections ; diagnosis ; Clostridium difficile ; genetics ; Electrophoresis, Capillary ; Feces ; microbiology ; Genes, Bacterial ; Humans ; Polymerase Chain Reaction ; Ribotyping ; Sensitivity and Specificity