Computational design of orthogonal membrane receptor-effector switches for rewiring signaling pathways.

Membrane receptors regulate numerous intracellular functions. However, the molecular underpinnings remain poorly understood because most receptors initiate multiple signaling pathways through distinct interaction interfaces that are structurally uncharacterized. We present an integrated computational and experimental approach to model and rationally engineer membrane receptor-intracellular protein systems signaling with novel pathway selectivity. We targeted the dopamine D2 receptor (D2), a G-protein-coupled receptor (GPCR), which primarily signals through Gi, but triggers also the Gq and beta-arrestin pathways. Using this approach, we designed orthogonal D2-Gi complexes, which coupled with high specificity and triggered exclusively the Gi-dependent signaling pathway. We also engineered an orthogonal chimeric D2-Gs/i complex that rewired D2 signaling from a Gi-mediated inhibitory into a Gs-dependent activating pathway. Reinterpreting the evolutionary history of GPCRs in light of the designed proteins, we uncovered an unforeseen hierarchical code of GPCR-G-protein coupling selectivity determinants. The results demonstrate that membrane receptor-cytosolic protein systems can be rationally engineered to regulate mammalian cellular functions. The method should prove useful for creating orthogonal molecular switches that redirect signals at the cell surface for cell-engineering applications.

[On-pump total arterial revascularization in coronary artery disease patients with left ventricular dysfunction: a multi-center retrospective study].

Objective: To evaluate the clinical outcomes of on-pump total arterial revascularization with bilateral radial artery (BRA) and left internal mammary artery (LIMA) as conduits in coronary artery bypass grafting (CABG) patients with left ventricular dysfunction (LVD). Methods: All the perioperative medical records and follow-up results of coronary artery disease patients with left ventricular ejection fraction (LVEF) ≤ 40% undergoing CABG from 24 heart centers of 15 provinces and autonomous regions in China between July 2015 and December 2019 were retrospectively analyzed. Results: A total of 87 consecutive patients (55 males and 32 females) underwent on-pump CABG with BRA and LIMA, with a mean age of (57.5±9.1) years old. There were 22 patients complicated with primary hypertension, 12 with diabetes mellitus, 8 with peripheral vascular disease, 7 with chronic obstructive lung disease, 12 with mild renal injury and 3 with partial aortic calcification. There were 43 cases with in-stent stenosis, and 21 had left main disease. The mean LVEF and left ventricular end-diastolic diameter (LVEDD) was (35.5±7.3)% and (65.5±2.6) mm, respectively. The mean graft number, aortic cross-clamp time and cardiopulmonary bypass duration was 3.2±0.9, (90.5±22.7) min and (113.4±19.2) min, respectively. There were 32 mitral and 9 aortic valve replacements, and 5 tricuspid annuloplasties. Prophylactic intra-aortic balloon pumps were implanted in 27 patients. There were 2 operative deaths from acute heart failure. After surgery, there were 15 cases of atrial fibrillation, 1 case of acute kidney injury, 1 case of acute myocardial infarction, and 1 cases of stroke. All the patients fulfilled the follow-up, with a mean time of (39.5±7.7) months. At 3 months after surgery, LVEDD was decreased and LVEF was improved significantly compared with pre-operative indicators [(53.0±1.5) mm vs (65.5±2.6) mm, t=9.51 P=0.02; (45.2±3.3)% vs (35.5±7.3)%, t=13.79, P=0.001]. No major cardiac events were reported during the follow-up. At (30.5±7.4) months after surgery, 62.4% of patients (53/85) underwent coronary CT angiography examination, and the results indicated that the graft patency was 98.8%, with only one case of RA occlusion occurred. Conclusion: In selected patients of LVD, on-pump total arterial revascularization with BRA and LIMA conduits was proved to be safe and effective.

[Pulsed electromagnetic fields stimulating osteogenic differentiation and maturation involves primary cilia-PI3K/AKT pathway].

OBJECTIVE: To study whether the pulsed electromagnetic fields (PEMF) promoting rat osteoblasts differentiation and maturation is related to the primary cilia and PI3K/AKT pathway, and to explore the mechanism of PEMF in promoting bone differentiation. METHODS: Enzyme solution was used to obtain newborn SD rats calvarial osteoblasts (ROB), which were processed by 50 Hz 0.6 mT PEMF for 0, 0.5, 1, 1.5 and 2 h, detecting PI3K and AKT protein expression and changes in primary cilia length and incidence; with LY294002 blocking PI3K/AKT signaling pathways we observed whether PEMF promoted osteogenic differentiation of ROB was affected; by interfering IFT88 gene expression by RNAi to inhibit primary cilia we observed whether PI3K/AKT signaling pathway and osteogenic differentiation of ROB was affected. Osteogenic differentiation indexes included alkaline phosphatase (ALP) activity, Real-time PCR and Western blot detection of osteogenic related genes of BMP-2, COL-1 and OSX and calcified nodules number, etc.. RESULTS: After exposure to PEMF for 0, 0.5, 1, 1.5, and 2 h, the protein expression of PI3K and AKT in ROB were increased significantly (P<0.01) and the primary cilia became longer; and the protein expression of PI3K reached the highest level at 0.5 h, as the treatment time of PEMF increased, the PI3K protein expression decreased. AKT showed higher protein expression at 0.5 h and 1.5 h. After blocking the PI3K/AKT signaling pathway with the PI3K blocker LY294002, PEMF could no longer increase ALP activity and the gene expressions of BMP-2, COL-1, OSX which were osteogenically related. However, PEMF could increase the ALP activity and the osteogenically related gene expression in ROB before blocking. After RNAi interfered the primary cilia, PEMF could no longer increase the protein expression of PI3K, which indicated that PEMF could not activate the PI3K/AKT signaling pathway after primary cilia interfering; secondly, the effect of PEMF on enhancing ALP activity disappeared, it also decrease the gene expressions of BMP-2, COL-1, and OSX, and the ability of increasing the calcification nodule formation also disappeared, indicating that the ability of PEMF to promote osteoblast maturation and mineralization disappeared after primary cilia interference. CONCLUSION: PEMF activated the PI3K/AKT signaling pathway through primary cilia on the surface of osteoblasts, then promoted bone formation activity and differentiation.

Exposure Duration Is a Determinant of the Effect of Sinusoidal Electromagnetic Fields on Peak Bone Mass of Young Rats.

We proposed a three-step strategy to obtain the optimal therapeutic parameters, which is composed of large-scale screening at cellular level, verification in animal experiments, and confirmation by a clinical trial. The objective of the current study was to test the feasibility of our strategy. Newborn rat calvarial osteoblasts were treated by 50 Hz 1.8 mT sinusoidal electromagnetic fields (SEMFs) with 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 h/days, respectively. The osteogenic differentiation and maturation of the osteoblast were assayed and compared to obtain the optimal duration. One-month-old growing rats were then treated by the same SEMFs with 0.5, 1.5, and 2.5 h/days, respectively, and the peak bone mass was analyzed after 2 months. It was found that the optimal exposure duration to promote the osteogenic differentiation and maturation of osteoblasts was 1.5 h/days, judging by the increasing degrees of ALP activity, calcified nodules formed, the gene and protein expression levels of Runx-2, BMP-2, and Col-I, as well as the expression levels of signaling proteins of the BMP-2/Smad1/5/8 pathway. The highest increase of peak bone mass after 2 months was also obtained by 1.5 h/days, judging by the results of X-ray dual-energy absorptiometry, mechanical property analysis, micro-CT scanning, and serum bone turnover marker examinations. The above results indicated that exposure duration is a determinant for the therapeutic effect of EMFs, and the optimal therapeutic effects only can be obtained by the optimal exposure duration.

Methylation markers differentiate thyroid cancer from benign nodules.

PURPOSE: The incidence of thyroid cancer (TC) is increasing. Cytology by itself cannot distinguish TC from some benign nodules especially in certain subtypes of TC. Our immediate goal is to identify DNA methylation markers for early detection of TC and to molecularly differentiate TC subtypes from benign nodules. METHODS: Promoter methylation status of 21 candidate genes was examined on formalin-fixed paraffin-embedded tissue (FFPE) utilizing quantitative methylation-specific polymerase chain reaction (QMSP) in a retrospective cohort of 329 patients (56% white, 29% African American, 61% female) comprising 71 normal thyroid, 83 benign nodules [follicular adenomas (FA)], 90 follicular TC (FTC) and 85 papillary TC (PTC). All genes were analyzed individually (Kruskal-Wallis and Wilcoxon rank sum tests) and in combination (logistic regression models) to identify genes whose methylation levels might best separate groups. RESULTS: Combination gene panels TPO and UCHL1 (ROC = 0.607, sensitivity 78%) discriminated FTC from FA, and RASSF1 and TPO (ROC = 0.881, sensitivity 78%) discriminated FTC from normal. Methylation of TSHR distinguished PTC from FTC (ROC = 0.701, sensitivity 84%) and PTC from FA (ROC = 0.685, sensitivity 70%). The six gene panel of TIMP3, RARB2, SERPINB5, RASSF1, TPO and TSHR, which differentiates PTC from normal thyroid, had the best combination sensitivity (91%) and specificity (81%) of the panels addressing discrimination of cancer tissue. CONCLUSIONS: Aberrant gene methylation used in combination panels may be useful clinically in differentiating FTC and PTC from benign nodules. If confirmed in additional studies, these findings could help reduce the over diagnosis of thyroid cancer and surgeries related to over diagnosis.

Primary cilium is required for the stimulating effect of icaritin on osteogenic differentiation and mineralization of osteoblasts in vitro.

OBJECTIVE: Icaritin, one effective metabolite of Herba Epimedii-derived flavonoid icariin, has a strong osteogenic activity. However, its action mechanism remains unclear. Since primary cilia have been shown to play a pivotal role in regulating the osteogenesis, we hypothesized primary cilia are indispensable in mediating icaritin osteogenic effect. MATERIALS AND METHODS: Primary rat calvarial osteoblasts were transfected with siRNA1 targeting intraflagellar transport protein 88 (IFT88), a protein required for ciliogenesis, to prevent formation of primary cilium and were treated with 10-6 M icaritin. RESULTS: Alkaline phosphatase (ALP) activity was significantly increased after 3 days in cells transfected with scrambled siRNA control and treated by icaritin (SC+I group) compared to cells transfected with scrambled siRNA control only (SC group). ALP activity after IFT88 siRNA1 transfection and icaritin treatment (siRNA1+I group) was significantly lower than that of SC+I group. Formation of ALP positively stained colonies after 6 days, osteocalcin secretion after 9 days and formation of calcified nodules after 12 days displayed a similar tendency among the three groups. mRNA expression of osteogenesis-related genes ALP, BMP-2, COL1α, RUNX-2 and OSX after 24 h was significantly increased in SC+I group, but was not different with SC group in siRNA1+I group. Protein levels of BMP-2, COL1α, RUNX-2 and OSX after 48 h showed the similar tendency with gene expression. CONCLUSION: Primary cilia are important in mediating icaritin-stimulated osteogenic differentiation and may be a novel target for pharmacological therapies for bone loss.

[Effect and related mechanism of microRNA-181 attenuates oxidized low density lipoprotein induced vascular endothelial cell injury].

Objective: To observe the expression level of microRNA-181 (miR-181) and importin-α3 in oxidized low density lipoprotein (ox-LDL) induced vascular endothelial cell injury models, and explore the effect and mechanism of miR-181 on endothelial cell injury. Methods: Human vein endothelial cell line CRL-1730 were cultured and vascular endothelial cell injury model was established by intervention with ox-LDL. The cells were divided into control group (intervened by double distilled water), low-dose group (intervened by 10 µg/ml ox-LDL) and high-dose group (intervened by 20 µg/ml ox-LDL). In addition, cells of low-dose group were divided into miR-181 mimic group (miR-181 mimic was transfected) and mimic control group (miR-181 mimic control was transfected). Cell viabilities, mRNA and protein expression level of interleukin-6 (IL-6), miR-181, importin-α3, and nuclear transcription factor-κB (NF-κB) were measured by methyl thiazolyl tetrazolium (MTT), real-time PCR and Western blot, respectively. Results: (1) The cell viabilities in low-dose group and high-dose group were lower than control group (0.207±0.012 and 0.204±0.007 vs. 0.323±0.018, all P<0.01). The relative IL-6 mRNA expression in low-dose group and high-dose group were higher than control group (1.24±0.16 and 1.36±0.23 vs. 0.22±0.03, all P<0.01). The relative miR-181 mRNA expression in low-dose group and high-dose group were lower than control group (0.91±0.11 and 0.88±0.07 vs. 2.20±0.13, all P<0.01). The relative importin-α3 mRNA expression in low-dose group and high-dose group were higher than control group (1.23±0.22 and 0.55±0.03 vs. 0.44±0.06, all P<0.01). The relative NF-κB mRNA expression in low-dose group and high-dose group were higher than control group (1.67±0.34 and 0.41±0.11 vs. 0.11±0.04, all P<0.01). The relative importin-α3 protein expression in low-dose group and high-dose group were higher than control group (1.44±0.23 and 1.31±0.22 vs. 0.29±0.08, all P<0.01). The relative NF-κB protein expression in low-dose group and high-dose group were higher than control group (0.43±0.05 and 0.37±0.04 vs. 0.16±0.03, all P<0.01). (2)The cell viabilities in miR-181 mimic group was higher than in mimic control group (0.262±0.008 vs. 0.211±0.021, P<0.01). The relative miR-181 mRNA expression level in miR-181 mimic group was higher than in mimic control group (4.23±0.34 vs. 0.88±0.16, P<0.01). The relative importin-α3 mRNA expression level in miR-181 mimic group was lower than in mimic control group (0.24±0.03 vs. 1.08±0.13, P<0.01). The relative NF-κB mRNA expression level was lower in miR-181 mimic group than in mimic control group (0.13±0.03 vs. 0.51±0.06, P<0.01). The relative importin-α3 protein expression level was lower in miR-181 mimic group than in mimic control group (0.34±0.06 vs. 1.67±0.26, P<0.01). The relative NF-κB protein expression level was lower in miR-181 mimic group than in mimic control group (0.43±0.02 vs. 1.53±0.36, P<0.01). Conclusions: Lower miR-181 expression but higher importin-α3 and its downstream NF-κB signaling are associated with ox-LDL induced vascular endothelial cell injury and up-regulation of miR-181 could alleviate ox-LDL induced vascular endothelial cell injury possibly via importin-α3/NF-κB pathway.

Differential diagnosis of pancreatic serous oligocystic adenoma and mucinous cystic neoplasm with spectral CT imaging: initial results.

AIM: To investigate the imaging characteristics of pancreatic serous oligocystic adenoma (SOA) and mucinous cystic neoplasms (MCNs) using spectral computed tomography (CT) and to evaluate whether quantitative information derived from spectral imaging can improve the differential diagnosis of these diseases. MATERIALS AND METHODS: From February 2010 to June 2013, 44 patients (24 SOAs and 20 MCNs) who underwent spectral CT imaging were included in the study. Conventional characteristics and quantitative parameters were compared between the two disease groups. Logistic regression was used for multiparametric analysis. The receiver-operating characteristic curve was used to evaluate the diagnostic performance of single parameter and multiparametric analysis. Two radiologists diagnosed the diseases blinded and independently, without and with the information of the statistical analysis. RESULTS: Tumour location, contour, size, and monochromatic CT values at 40 keV to 70 keV, iodine concentration, and effective atomic number (effective-Z) in the late arterial phase were the independent factors correlated with category. Multiparametric analysis with logistic regression showed that tumour size, location, and contour were the most effective variations, and obtained an area under the ROC curve (AUC) of 0.934. With the knowledge of statistical analysis, the accuracy of the first reader increased from 70.5% to 86.4%, and the accuracy of the second reader increased from 81.8% to 90.9%. CONCLUSIONS: Although CT spectral imaging provided additional information and multiparametric analysis obtained better performance than single-parameter analysis in differentiating MCNs from SOAs, multiparametric analysis with the combination of quantitative parameters derived from CT spectral imaging did not improve the diagnostic performance. Tumour size, location, and contour played an important role in differentiating MCNs from SOAs.

Evaluation of coronary artery in-stent restenosis with prospectively ECG-triggered axial CT angiography versus retrospective technique: a phantom study.

PURPOSE: This study compared the performance of prospectively electrocardiographically (ECG)-triggered axial computed tomography (CT) angiography with retrospective technique in evaluating coronary artery stent restenosis by 64-slice CT. MATERIALS AND METHODS: A pulsing cardiac phantom with artificial coronary artery in-stent restenosis was examined by CT angiography with different types of scan modes. The visibility of in-stent restenosis was evaluated with a three-point score. Artificial lumen narrowing [(inner stent diameter-measured lumen diameter)/inner stent diameter], lumen attenuation increase ratio [(in-stent attenuation-coronary artery lumen attenuation)/coronary artery lumen attenuation], measurement error of restenosis percent [(known restenosis percent-measured restenosis percent)/known restenosis percent] and imaging noise were analysed. RESULTS: Prospective acquisition showed better visibility than retrospective acquisition (p<0.05): 61% of in-stent restenoses had good visibility on the prospective acquisition compared with 17% on the retrospective acquisition. Furthermore, the effective dose was 6.2 ± 0.3 mSv for the prospective technique compared with 18.8 ± 1.1 mSv for the retrospective technique. Artificial lumen narrowing (mean 40%), lumen attenuation increase ratio (mean 33%) and measurement error of restenosis percent were not different between types of CT acquisitions. CONCLUSIONS: Compared with the traditional retrospective technique, prospective coronary CT angiography offers improved image quality and reduces effective radiation dose in evaluating in-stent restenosis.

FlowCytomix analysis for Toxoplasma gondii infection in pregnant women in central Taiwan.

The study was to determine the seroprevalence of toxoplasmosis in the sera of pregnant women in central Taiwan and to investigate the levels of cytokine in the sera of pregnant women with Toxoplasma gondii infection. The 220 blood samples were collected from pregnant women. The haematological parameters of peripheral blood were analysed by a haematology analyser. Serum samples of the pregnant women were analysed by a commercially available anti-T. gondii IgM/IgG antibody enzyme-linked immunosorbent assay (ELISA) kit and FlowCytomix assays. Six (2.7%) of the sera samples had IgM anti-T. gondii antibodies, and twenty (9.1%) had T. gondii IgG seropositive. All six IgM seropositive samples had low IgG avidity, indicative of acute infection. Total white blood cells and eosinophils were statistically significantly increased (p<0.05) in pregnant women with T. gondii infection, as compared with healthy pregnant women. Th1 cytokines IFN-γ, IL-1ß, IL-2 and IL-12 p70, and Th2 cytokines IL-10 in pregnant women with T. gondii IgM/IgG seropositive were significantly increased (p<0.05), as compared with healthy pregnant women. These results showed that both of Th1 and Th2 cytokines play an important role in the toxoplasmosis of pregnant women.

Induction of plasminogen activators in pregnant women with Toxoplasma gondii infection.

OBJECTIVE: To determine whether plasminogen activators (PAs) are involved in the pathologic process of toxoplasmosis. MATERIALS AND METHODS: Out of 220 pregnant women the study included 26 with a diagnosis of toxoplasmosis: six based on seropositivity for Toxoplasma gondii IgM and 20 based on seropositivity for T. gondii IgG. We measured serum activities and protein levels of PAs by casein zymography and Western blotting, respectively. RESULTS: Serum PAs were higher in healthy pregnant women than in their healthy nonpregnant counterparts. Furthermore, serum PAs were significantly higher in pregnant women infected with T. gondii than in their healthy counterparts. CONCLUSION: PAs participate in the pathogenesis of toxoplasmosis in pregnant women and may be useful markers of T. gondii infection.

MiR-101 affects proliferation and apoptosis of cervical cancer cells by inhibition of JAK2.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-101 affects proliferation and apoptosis of cervical cancer cells by inhibition of JAK2, by H. Wei, W.-R. He, K.-M. Chen, X.-W. Wang, C.-J. Yi, published in Eur Rev Med Pharmacol Sci 2019; 23 (13): 5640-5647-DOI: 10.26355/eurrev_201907_18299-PMID: 31298355" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18299.

Prospective study comparing two iodine concentrations for multidetector computed tomography of the pancreas.

PURPOSE: The authors sought to determine the influence of two different iodine concentrations of nonionic contrast media (cm) on contrast enhancement in pancreatic computed tomography angiography (CTA). MATERIALS AND METHODS: Sixty patients with clinically suspected or known pancreatic disease underwent pancreatic CTA. The patients were randomly assigned to group A (n = 30) and group B (n = 30). The contrast agent was injected with iodine concentrations of 400 mg I/ml (Iomeron 400) in group A and 300 mg I/ml (Iopamidol 300) in group B with the same total iodine dose (36 g). Arterial and portal venous phase contrast enhancement of the vessels, organs and pancreatic masses was measured, and blinded qualitative image assessment was performed by two expert radiologists. RESULTS: In the arterial and portal venous phase, the highly concentrated cm led to significantly greater enhancement in the abdominal main vessels, pancreas and pancreatic carcinoma than did the low concentrated cm. No statistically significant attenuation differences were measured between pancreatic carcinomas and the pancreatic parenchyma in the arterial and portal venous phase between group A and B. The overall trend for both readers was to assign higher scores to group A than group B. CONCLUSIONS: The higher iodine concentration leads to greater contrast enhancement of abdominal vessels and organs in pancreatic CTA. Detection and demarcation of hypovascular pancreatic carcinoma was not found to be improved by the higher iodine concentration.

Cumulative survival in early-onset unilateral and bilateral breast cancer: an analysis of 1907 Taiwanese women.

As the epidemiological pattern of breast cancer in modernising Asian countries differs greatly from that in Western countries, it is worthwhile to investigate the long-term prognoses of unilateral and bilateral breast cancer in these nations. A retrospective cohort study composed of 1907 Taiwanese women was conducted to follow 1863 unilateral and 44 bilateral cases of breast cancer. Time-dependent Cox regression was used to assess the risk of breast cancer death by considering the time course of unilateral and bilateral tumour development. The 15-year survival rates were 68.37, 62.63, and 26.42% for unilateral, synchronous bilateral, and metachronous bilateral breast cancer, respectively. Differences among types were most apparent after 5 years of follow-up. After adjusting for significant prognostic factors, the risk of death for overall bilateral breast cancer was 2.50-fold greater (95% CI, 1.43-4.37) compared to unilateral breast cancer. The corresponding figures were 1.12-fold (95% CI, 0.42-3.02) and 6.11-fold (95% CI, 3.14-11.89) for synchronous and metachronous bilateral breast cancer, respectively. Taiwanese women, who are frequently diagnosed with breast cancer before 50 years of age, showed poorer survival for metachronous bilateral than for synchronous bilateral or unilateral breast cancer. Survival was markedly poorer compared to recent data from Sweden.

Crack-like sources of dislocation nucleation and multiplication in thin films.

With the combination of the height sensitivity of atomic force microscopy and the strain sensitivity of transmission electron microscopy, it is shown that near singular stress concentrations can develop naturally in strained epitaxial films. These crack-like instabilities are identified as the sources of dislocation nucleation and multiplication in films of high misfit. This link between morphological instability and dislocation nucleation provides a method for studying the basic micromechanisms that determine the strength and mechanical properties of materials.

The value of multidetector-row CT in the preoperative detection of pancreatic insulinomas.

PURPOSE: The authors sought to quantitatively analyse enhancement characteristics of pancreatic insulinomas in different phases and determine the value of multidetector-row computed tomography (CT) for detecting insulinomas. MATERIALS AND METHODS: Forty-six patients with surgically proven insulinomas diagnosed between 2002 and 2007 were retrospectively reviewed. These patients underwent single-phase (group 1) or dual-phase (group 2) helical CT scanning. RESULTS: Sensitivity for detecting insulinomas in group 2 was superior to that in group 1 (p<0.05).The sensitivity for insulinoma detection in the arterial phase was superior to that in the portal-venous phase (p<0.05). The mean attenuation values of the insulinomas and normal pancreas during the unenhanced arterial and portal-venous phases were, respectively, 40.5+/-8.75 HU (Hounsfield units), 114.48+/-27.30 HU, 112.19+/-19.52 HU and 44.56+/-6.48 HU, 81.16+/-15.22 HU, 90.54+/-13.80 HU, and there was statistical difference between them (p=0.000). The contrast enhancement of insulinomas in the arterial and portal-venous phases was 74.03+/-29.51 HU and 70.90+/-21.93 HU, respectively, and there was no statistical difference between them (p=0.499). The tumour to normal-pancreas attenuation differences in the arterial and portal-venous phases were respectively 33.32+/-20.96 HU and 20.58+/-16.32 HU, respectively, and there was statistical difference between them (p=0.011). CONCLUSIONS: Dual-phase CT has a promising sensitivity in detecting pancreatic insulinomas. The acquisition of images in the arterial phase is more helpful for detecting insulinomas.

MiR-101 affects proliferation and apoptosis of cervical cancer cells by inhibition of JAK2.

OBJECTIVE: JAK2 expression and dysfunction play a role in tumor pathogenesis. Bioinformatics analysis revealed a targeted binding site between miR-101 and the 3'-UTR of JAK2 mRNA. This study investigated the role of miR-101 in regulating JAK2 expression and affecting the proliferation and apoptosis of cervical cancer cells. PATIENTS AND METHODS: The tumor tissues and adjacent tissues of patients with cervical cancer were collected. The expression of miR-101 and JAK2 was detected by qRT-PCR. The dual luciferase reporter gene assay validated the targeting relationship between miR-101 and JAK2. The cervical cancer Caski cells were cultured in vitro, and divided into miR-NC group and miR-101 mimic group. The expression of JAK2 and p-JAK2 was detected by Western blot, cell apoptosis was detected by flow cytometry, and cell proliferation was detected by EdU staining. RESULTS: Compared with adjacent tissues, miR-101 expression was significantly decreased, and JAK2 expression was increased in cervical cancer tissues. There was a targeted regulatory relationship between miR-101 and JAK2. Compared with HcerEpic cells, miR-101 expression in HeLa and Caski was significantly decreased, and the expression of JAK2 and p-JAK2 was significantly increased. Transfection of miR-101 mimic significantly reduced the expression of JAK2 and p-JAK2 in Caski cells, reduced cell proliferation and increased cell apoptosis. CONCLUSIONS: The decrease of miR-101 expression and the increase of JAK2 expression play a role in cervical cancer, while the increase of miR-101 expression can inhibit the proliferation and promote the apoptosis of cells by inhibiting the expression of JAK2.

Analysis of gene mutation associated with tyrosine kinase inhibitor sensitivity of epidermal growth factor receptor in cervical cancer patients.

OBJECTIVE: Cervical cancer is frequent in females. Epidermal growth factor receptor has a prominent expression in certain malignant tumors. This study aims to observe the expressional profile of epidermal growth factor receptor (EGFR) in cervical cancer patients, and mutation of EGFR gene related with its sensitivity towards tyrosine kinase inhibitor. MATERIALS AND METHODS: Cervical cancer patients from our hospital were recruited as the experimental group, in parallel with chronic cervicitis patients as control group. Serum EGFR level was measured by enzyme-linked immunosorbent assay (ELISA), and EGFR levels in cervical tissues were quantified by immunohistochemistry assay (IHC) staining. Real Time-PCR (RT-PCR) examined mutations of exon 18, 19, and 21 of the EGFR gene, to analyze their correlation with clinical or pathological features. RESULTS: Serum EGFR in experimental group was 1.16 ± 0.04 ng/ml, significantly higher than control group (p < 0.05). EGFR positive rate was 71.1% in cancer tissues, significantly higher compared to controlled or adjacent tissues (p < 0.05). Mutation rats of EGFR exon 19 and exon 21 were 3.3% and 5%, respectively. No mutation was found in exon 18. Such mutations of EGFR gene were related with cancer differentiation grade, tumor-lymph-node-metastasis (TNM) stage, lymph node or distal metastasis (p < 0.05), but not age, Karnofsky performance score (KPS) score or infiltration depth. CONCLUSIONS: EGFR is highly expressed in serum and tumors of cervical cancer patients, some of which showed mutations of exon 19 and 21 of EGFR gene with relatively lower frequency. Mutation rates were significantly higher in patients with highly differentiated grade, early TNM stage, and those without lymph node or distal metastasis.

Changes to plasminogen activators and matrix metalloproteinase-9 in dogs with toxocarosis.

The relationship between proteolytic enzymes and hematological response to infection was studied in five 1-month-old dogs inoculated experimentally with 2000 eggs of Toxocara canis. Moderate leukocytosis and marked eosinophilia occurred 14 days post-infection with T. canis. Plasminogen activators (PAs) and matrix metalloproteinase-9 (MMP-9) activity in serum was significantly different in dogs infected with T. canis, compared with controls. Urokinase-type PA (uPA) activity was positively correlated with eosinophilia, and tissue-type PA (tPA) and MMP-9 activity was negatively correlated with eosinophilia. However, there was no correlation between inflammation and MMP-2. The use of uPA, tPA or MMP-9 proteolytic enzymes as laboratory reference markers for toxocarosis requires further study.

Icariin enhances the osteogenic differentiation of bone marrow stromal cells but has no effects on the differentiation of newborn calvarial osteoblasts of rats.

Since the total flavonoid extract (TFE) of Epimedium herb was found to prevent osteoporosis induced by ovariectomy in rats, we have been attempting to identify the exact compound responsible for the bone-strengthening activity. In this experiment, four flavonoid extracts were obtained from Epimedium sagittatum (Siebold & Zucc.) Maxim, which contained 25.3%, 51.2%, 82.3% and 99.2% icariin respectively. They were separately supplemented into the culture media of newborn rat calvarial osteoblasts (ROB) or primary rat bone marrow stroma cells (rMSCs) at 0.1, 1, 10 and 100 microg/ml respectively, in order to observe their effects on the cells. Not any appreciable effect was found on the differentiation of ROB, but an enhancing effect on the osteogenic differentiation of rMSCs was found, and the enhancing degree was icariin-dependent, that is, a higher concentration of icariin in the extract caused more mineralized bone nodules and higher calcium deposition levels. The gene expressions involved in osteogenesis were also improved which was revealed by RT-PCR, including alkaline phosphatase, bone matrix protein (osteocalcin, osteopontin, bone sialoprotein) and cytokines (TGF-beta1 and IGF-I). The effect of icariin on cell proliferation was assayed by the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Icariin inhibited the proliferation of rMSCs and ROB when its concentration was higher than 10(-5) microM (6.7 microg/ml), no stimulative effect was found. The above results indicated that icariin may exert bone-strengthening activity by enhancing the osteogenic differentiation of MSCs, which partially explains the anti-osteoporosis action of Epimedium herb.