Changes in circadian gene cryptochrome 2 expression in mouse models of psoriasis and HaCaT cells and their underlying mechanisms / 中华皮肤科杂志

Objective:To investigate changes in circadian gene cryptochrome 2 (CRY2) expression in mouse models of psoriasis and HaCaT cells, and to explore underlying mechanisms.Methods:Imiquimod-induced mouse model experiment: 12 C57BL/6 female mice were randomly and equally divided into imiquimod group receiving topical imiquimod treatment for 5 consecutive days and control group receiving no treatment; these mice were sacrificed on day 6, skin tissues were resected from the back of mice, and immunofluorescence staining was performed to determine the CRY2 expression in the epidermis. HaCaT cell transfection experiment: HaCaT cells with small interfering RNA (siRNA) -mediated knockdown of CRY2 served as siRNA-CRY2 group, and siRNA-NC group as control group; 5-ethynyl-2′-deoxyuridine (EdU) staining was performed to evaluate the proliferative activity of the HaCaT cells, real-time fluorescence-based quantitative PCR (qPCR) to determine the mRNA expression of chemokines in the HaCaT cells, and Western blot analysis to determine phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2) . Tumor necrosis factor-α (TNF-α) -stimulated animal and cell experiments: 12 C57BL/6 female mice were randomly and equally divided into TNF-α group subcutaneously injected with TNF-α solution in the ear for 6 days, and phosphate buffered saline (PBS) group subcutaneously injected with the same amount of PBS; the mice were sacrificed on day 7, skin tissues were resected from the ear of mice, and immunofluorescence staining was conducted to determine the CRY2 expression in the epidermis; CRY2-knockdown HaCaT cells stimulated with 50 ng/ml TNF-α for 12 hours served as siRNA-CRY2 + TNF-α group, and siRNA-NC + TNF-α group as control group; qPCR was performed to determine the mRNA expression of chemokines in HaCaT cells in the above groups. Statistical analysis was carried out by using two-independent-sample t test. Results:Immunofluorescence staining showed that the CRY2 protein expression was significantly lower in the mouse dorsal epidermis in the imiquimod group (0.94 ± 0.23) than in the control group (2.30 ± 0.25, t = 3.99, P = 0.016) . Compared with the siRNA-NC group, the siRNA-CRY2 group showed significantly increased proportions of EdU-positive cells (48.13% ± 10.97% vs. 38.23% ± 0.81%, t = 5.00, P = 0.007) , mRNA expression levels of chemokines CXCL1 and CXCL8, as well as significantly increased phosphorylated (p) -ERK1/2 protein expression levels (all P < 0.05) , while there were no significant differences in the CCL20 mRNA expression or ERK1/2 protein expression between the two groups (both P > 0.05) . Immunofluorescence staining showed significantly decreased CRY2 protein expression level in the mouse ear epidermis in the TNF-α group (0.37 ± 0.34) compared with the PBS group (2.04 ± 0.17, t = 4.38, P = 0.012) ; the relative mRNA expression levels of chemokines CXCL1, CXCL8, and CCL20 in HaCaT cells were significantly higher in the siRNA-CRY2 + TNF-α group than in the siRNA-NC + TNF-α group (all P < 0.05) . Conclusion:CRY2 was markedly underexpressed in psoriasis, which might promote the proliferation of keratinocytes and expression of chemokines CXCL1, CXCL8 and CCL20, and TNF-α might be an upstream cytokine that could downregulate CRY2 expression.

Pancreatic cancer cell-secreted exosomes promote apoptosis of β cells via activation of mitochondrial apoptotic pathways / 中国病理生理杂志

AIM:To investigate the effects of exosomes secreted by pancreatic cancer cells on the viability and function of βcells and the possible mechanism .METHODS:ExoQuick-TC kit was used to extract exosomes in the super-natants of mouse pancreatic cancer Pan 02 and MPC-83 cells, and the extracted exosomes were identified by transmission electron microscopy.Fluorescence-labeled exosomes were incubated with mouse insulinoma MIN 6 cells for 48 h to detect whether exosomes secreted by pancreatic cancer cells were uptaken by MIN 6 cells.MTT and glucose-stimulated insulin se-cretion ( GSIS) assays were conducted to examine cell viability and insulin secretion of MIN 6 cells after incubating with ex-osomes.The expression of miR-204 and Bcl-2 mRNA in MIN6 cells was detected by qPCR .The protein expression of Bcl-2, Bax, caspase-3 and cytochrome C (Cyt-C) in MIN6 cells was determined by Western blot .RESULTS:The results of transmission electron microscopy showed that both Pan 02 cells and MPC-83 cells secreted exosomes , and Pan02 cells secre-ted more.The co-incubation results of fluorescence-labeled exosomes and MIN6 cells confirmed that MIN6 cells were able to ingest large amounts of exosomes secreted by pancreatic cancer cells .The results of MTT and GSIS assays showed that the viability and the level of high glucose-stimulated insulin secretion of MIN 6 cells in exosome treatment group significantly decreased compared with nontreatment group (P<0.01).The results of qPCR showed that the exosomes secreted by pan-creatic cancer cells were rich in miR-204, and the mRNA expression of Bcl-2 in MIN6 cells was significantly down-regula-ted by exosome incubation ( P<0.01) .The results of Western blot showed that the protein expression of Bcl-2 in the MIN6 cells treated with exosomes was significantly down-regulated (P<0.05), and the protein levels of Bax, cleaved caspase-3 and Cyt-C in exosomes treatment group were significantly up-regulated ( P<0.01 ) .CONCLUSION: Pancreatic cancer cells secrete exosomes .The exosomes secreted by pancreatic cancer cells are ingested by βcells, and reduce the viability and insulin secretion of βcells.The mechanism may be related to the increase in exosomal miR-204 in the βcells.In-creasing miR-204 may inhibit the expression of Bcl-2 and promote the activation of mitochondrial apoptosis in βcells.

Focusing on MRI-suspected lesions in targeted transrectal prostate biopsy guided by MRI-TRUS fusion imaging for the diagnosis of prostate cancer / 中华男科学杂志

<p><b>Objective</b>To improve the accuracy of prostate cancer (PCa) detection by focusing biopsy on the suspected lesion manifested by MRI with the total number of biopsy cores relatively unchanged.</p><p><b>METHODS</b>A prospective randomized analysis was performed on 262 cases of suspected PCa detected by multi-parametric MRI (mp-MRI), each with a single suspected lesion with 10 μg/L≤ PSA <20 μg/L. All the patients underwent targeted transrectal prostate biopsy guided by fusion imaging of MRI with transrectal ultrasonography (TRUS), using the 6X+6 strategy (6 cores in the suspected region and another 6 in the systematic prostate) for 134 cases and the traditional 12+2X method (12 cores in the systematic prostate and 2 in the suspected region) for the other 128. Comparisons were made between the two methods in the PCa detection rate in the cases of suspected lesion, total PCa detection rate, incidence of post-biopsy complications, and Gleason scores. Analyses were performed on the prostate imaging reporting and data system (PI-RADS) score, location, transverse section, and diameter of the suspected lesion.</p><p><b>RESULTS</b>Both the total PCa detection rate and that in the cases of suspected lesion were significantly higher in the 6X+6 (44.8% and 37.3%) than in the 12+2X group (37.5% and 27.3%) (P<0.05). MRI showed that the suspected lesions were mostly (45%) located in the middle part of the prostate, the mean area of the transverse section was (0.48±0.11) cm2, and the mean diameter of the tumor was (8.51±2.21) mm. The results of biopsy showed that low-grade tumors (Gleason 3+3=6) accounted for 68% in the 6X+6 group and 71% in the 12+2X group. No statistically significant differences were found between the two groups in the incidence rate of post-biopsy complications.</p><p><b>CONCLUSIONS</b>Compared with the traditional 12+2X method, for the suspected lesion manifested by mp-MRI, focusing biopsy on the suspected region with the 6X+6 strategy can achieve a higher PCa detection rate without increasing the incidence of complications.</p>

Factors Affecting the Accuracy of Intraocular Lens Power Calculation with Lenstar / 전남의대학술지

This retrospective study was performed to compare refractive outcomes measured by conventional methods and by use of the Lenstar biometer and to investigate the factors affecting intraocular lens (IOL) power calculation with Lenstar with and without IOL-constant optimization. The study included 100 eyes of 86 patients who underwent cataract surgery. Corneal curvature was measured with a manual keratometer (MK), automated keratometer (AK), and the Lenstar biometer, and axial length (AL) was measured by A-scan and Lenstar. Mean numerical error (MNE) and mean absolute error (MAE) were compared between AK and MK with A-scan, and Lenstar with and without optimization. Factors affecting the accuracy of the IOL power calculation by use of Lenstar with and without optimization were analyzed. No significant differences were observed in the MNE or MAE among the devices. The proportion of MAE within 0.5 D was higher for Lenstar with optimization (62.7%) than without optimization (46.2%). The proportion of MAE within 0.5 D was 62% and 58% for MK and AK with A-scan, respectively. Without optimization, the MAE was smaller in eyes with ALs between 23 mm and 25 mm (p=0.03), whereas it was smaller at higher corneal powers when the IOL constant was optimized (>44 D, p=0.03). The IOL power calculations showed no significant differences among the devices, but the results of MAE within 0.5 D by use of Lenstar without optimization were worse than those of conventional methods. The AL influenced the accuracy of refractive outcomes determined by using Lenstar without optimization, and corneal curvature was shown to affect the accuracy of refractive measurements using Lenstar with optimization.

Factors Affecting the Accuracy of Intraocular Lens Power Calculation with Lenstar / 전남의대학술지

This retrospective study was performed to compare refractive outcomes measured by conventional methods and by use of the Lenstar biometer and to investigate the factors affecting intraocular lens (IOL) power calculation with Lenstar with and without IOL-constant optimization. The study included 100 eyes of 86 patients who underwent cataract surgery. Corneal curvature was measured with a manual keratometer (MK), automated keratometer (AK), and the Lenstar biometer, and axial length (AL) was measured by A-scan and Lenstar. Mean numerical error (MNE) and mean absolute error (MAE) were compared between AK and MK with A-scan, and Lenstar with and without optimization. Factors affecting the accuracy of the IOL power calculation by use of Lenstar with and without optimization were analyzed. No significant differences were observed in the MNE or MAE among the devices. The proportion of MAE within 0.5 D was higher for Lenstar with optimization (62.7%) than without optimization (46.2%). The proportion of MAE within 0.5 D was 62% and 58% for MK and AK with A-scan, respectively. Without optimization, the MAE was smaller in eyes with ALs between 23 mm and 25 mm (p=0.03), whereas it was smaller at higher corneal powers when the IOL constant was optimized (>44 D, p=0.03). The IOL power calculations showed no significant differences among the devices, but the results of MAE within 0.5 D by use of Lenstar without optimization were worse than those of conventional methods. The AL influenced the accuracy of refractive outcomes determined by using Lenstar without optimization, and corneal curvature was shown to affect the accuracy of refractive measurements using Lenstar with optimization.

Determination of flavone for Scutellaria baicalensis from different areas by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To describe the difference between native and nonative herbs by determining contents of seven kinds of flavone for twenty-five samples from seventeen areas.</p><p><b>METHODS</b>HPLC. Fluid phase: MEOH-H2O-CH3COOH(ICE) (41:59:0.2) and (50:50:0.2). Detection wavelength: 275.</p><p><b>RESULTS</b>The contents of baicalin are 6%-9%, wogenin are 2%-8%, baicalein are 0.1%-1.6%, neobaicalein are 0.01%-0.2%, wogonin are 0.01%-0.3%, visidulin and oroxylin are trace amounts or undetected.</p><p><b>CONCLUSION</b>The native and nonative herbs have no distinct differce in absolute component ratio. The ratio of baicalin and wogenin is under three. The ratio of baicalin and baicalein, baicalin and wogonin is between twenty and fifty.</p>