Relationships between aneurysmal wall enhancement and conventional risk factors in patients with intracranial aneurysm: A high-resolution MRI study.

BACKGROUND AND PURPOSE: Aneurysmal wall enhancement (AWE) is thought to reflect wall inflammation and is a novel imaging biomarker for intracranial aneurysm (IA) risk evaluation. However, the relationship between AWE and other conventional risk factors (e.g., size) remains unclear. The aim of this study was to investigate the relationship between AWE and other risk factors. MATERIAL AND METHODS: Seventy-six consecutive patients from February 2016 to April 2017 with 88 unruptured IAs were reviewed. Patients and IAs were divided into with AWE and without AWE groups according to high-resolution magnetic resonance imaging (HRMRI) images. In addition to the patients' clinical characteristics, the features of the IAs (e.g., size and aspect ratio (AR)) were evaluated via computed tomography angiography. Multiple logistic regression analysis was used to identify the association between AWE and other risk factors. A receiver operating characteristic curve analysis was performed for the final model to obtain optimal thresholds. RESULTS: IAs with an irregular shape (OR 12.544) and a high AR (OR 32.891) were associated with AWE. The threshold value of the AR was 1.05. CONCLUSIONS: AWE on contrast-enhanced HRMRI was correlated with IAs with an irregular shape and a high AR. AWE may be a marker of instability and even risk of rupture.

[Efficacy and safety of № I Empirical Prescription for Chronic Prostatitis in the treatment of type ⠢ refractory chronic prostatitis].

OBJECTIVE: To evaluate the efficiency and safety of № I Empirical Prescription for Chronic Prostatitis (№ I EPCP) in the treatment of type Ⅲ refractory chronic prostatitis. METHODS: We randomly assigned 53 cases of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis to an experimental and a control group to receive № I EPCP at 1 dose per day and saw palmetto extract at 160 mg bid), respectively, all for 8 weeks. Before and after 4 and 8 weeks of treatment, we obtained The National Institute of Health Chronic Prostatitis Symptom Index (NIH-CPSI) scores, Traditional Chinese Medicine Syndrome Scores (TCMSS), maximum urinary flow rate (Qmax), average urinary flow rate (Qavg), Hamilton Depression Rating Scale (HAMD) scores and Hamilton Anxiety Rating Scale (HAMA) scores, and compared them between the two groups of patients. RESULTS: Totally 48 of the patients completed the medication and follow-up, 25 in the experimental and 23 in the control group. Compared with the baseline, the NIH-CPSI scores after 8 weeks of treatment were significantly decreased in the experimental (27.82 ± 7.25 vs 15.46 ± 4.77, P <0.05) and the control group (25.98 ± 6.47 vs 21.06 ± 5.74, P <0.05), and so were the TCMSSs (24.64 ± 9.82 vs 16.42 ± 6.33 and 9.15 ± 3.74, P <0.05, and 23.67 ± 8.73 vs 18.55 ± 5.92 and 13.48 ± 4.45, P <0.05); the Qmax at 8 weeks were dramatically increased in the experimental group (ï¼»18.45 ± 7.81ï¼½ vs ï¼»23.44 ± 8.73ï¼½ ml/s, P <0.05) and the control (ï¼»17.58 ± 6.92ï¼½ vs ï¼»21.26 ± 8.32ï¼½ ml/s, P <0.05), and so was the Qavg (ï¼»11.27 ± 5.33ï¼½ vs ï¼»16.51 ± 7.36ï¼½ ml/s, P <0.05 and ï¼»10.66 ± 5.82ï¼½ vs ï¼»13.44 ± 6.16ï¼½ ml/s, P <0.05); the HAMD scores were remarkably reduced in the experimental group (22.74 ± 6.37 vs 17.62 ± 5.71 and 12.54 ± 5.22, P <0.05) and the control (23.55 ± 7.14 vs 22.34 ± 6.88 and 21.62 ± 5.63, P <0.05), and so were the HAMA scores (21.37 ± 7.15 vs 18.42 ± 6.35 and 14.63 ± 7.11, P <0.05 and 20.54 ± 6.77 vs 19.87 ± 6.24 and 19.42 ± 7.04, P <0.05). No obvious adverse reactions were observed in either of the two groups during the medication. CONCLUSIONS: № I EPCP deserves promotion and clinical application for its definite effectiveness and safety in the treatment of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis.

[Pharmacokinetics and MR imaging of SPIO-shRNA dual functional molecular probe in vivo].

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.

[Optimal concentration of superparamagnetic iron oxide-short hairpin RNA dual functional molecular probe transfected into ovarian cancer cells in vitro].

OBJECTIVE: To explore the effects of superparamagnetic iron oxide-short hairpin RNA (SPIO-ShRNA) dual functional molecular probes of different concentrations on morphology and biological behavior of ovarian cancer SKOV3 cells in vitro. METHODS: The dual functional molecular probes at an iron concentration of 5, 15, 30, 45, 75, and 100 mg/L were transfected into SKOV3 cells. The transfection rate of the probe was observed by fluorescence microscope. The distribution and content of iron particles in SKOV3 cells were determined by Prussian blue staining, atomic adsorption spectrometer and electron microscopy. Cell viability was observed by cell counting kit-8 (CCK-8).The apoptosis was detected by flow cytometry. The expression of protein within the cells was detected by Western blot. The changes of the signal intensity were measured by magnetic resonance imaging (MRI). RESULTS: The SPIO-ShRNA dual functional molecular probe was uptaken in aconcentration-dependence manner within a certain range (5-30 mg/L). When the concentration of the probe was 45 mg/L, the labeling rate of the cell was close to 100%; With the increase of the concentration of probe, the cell survival rate decreased gradually. The cell survival rate of each experimental group were 94.626%±1.050%, 93.373%±1.180%, 91.700%±3.122%, 75.100%±4.362%, 72.983%±3.233%, 71.010%±2.910%,5, 15, 30 mg/L cell survival rate was not significantly decreased, the difference was not statistically significant (P=0.226, P=0.068, P=0.475); When the concentration of the probe was greater than or equal to 45 mg/L,the survival rate decreased obviously (P<0.001); Group of 45 mg/L protein expression rate was 68.905%±3.510%, When the concentration of the probe was greater than or equal to 45 mg/L, the inhibition rate of the protein expression level of epidermal growth factor receptor was obviously higher than those of 5, 15, and 30 mg/L groups, the difference was statistically significant (P<0.001, P=0.001, P=0.003, all P<0.01); the MRI displayed that the signal intensity was decreased with increasing concentrations of the probe. The signal intensity of 45 mg/L group was 165.55±4.92, compared with the blank control group(same volume of phosphate buffer saline), normal group (unlabeled ovarian cancer SKOV3 cells), 5, 15, and 30 mg/L groups , the signal intensity of 45 mg/L group decreased significantly (all P<0.001). CONCLUSION: The dual functional molecular probe can effectively transfect and specifically inhibit the expression of SKOV3 cell lines at the iron concentration of 45 mg/L, and can also be detected by MRI. The role of diagnosis and treatment of the dual functional molecular probe has been initially confirmed.

Effects of external magnetic field on the transfection rate of SPIO-shRNADual functional molecular probe into ovarian carcinoma SKOV3 cells in vitro.

OBJECTIVE: To explore the transfection rate of SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells in external magnetic field. METHODS: Dual functional molecular probe at an iron concentration of 45 mg/L was transfected into SKOV3 cells. The cells with coexisting probe and magnetic fields were set as the intervention group,the probe-transfected cells as negative control group, and normally cultured SKOV3 without any transfection as blank control group. The transfection rate was detected by flow cytometry. Cell viability was observed by CCK-8 assay. Epidermal growth factor receptor (EGFR) expression level in SKOV3 cells was determined by real-time quantitative PCR and Western blot analysis. The signal intensity was measured by magnetic resonance imaging (MRI). RESULTS: The transfection rate of the intervention group was (79.20 ± 3.31)%, which was significantly higher than that of negative control group (P=0.001). Compared with the negative control group,the cell viability of the intervention group significantly decreased (P=0.011), protein and mRNA expression levels of EGFR in the intervention group were significantly decreased (both P<0.05). The signal intensity on T2(*)WI in the intervention group also significantly decreased (P=0.0004). CONCLUSION: The external magnetic field can improve the transfection efficiency SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells.

MRCP Combined With CT Promotes the Differentiation of Benign and Malignant Distal Bile Duct Strictures.

OBJECTIVE: To determine whether contrast-enhanced computed tomography (CT) can promote the identification of malignant and benign distal biliary strictures (DBSs) compared to the use of magnetic resonance cholangiopancreatography (MRCP) alone and to identify imaging findings of malignant DBSs. MATERIALS AND METHODS: A total of 168 consecutive patients with confirmed DBSs were reviewed. MRCP alone and MRCP combined with CT images were blindly analyzed by two radiologists (e.g., stricture pattern, margins), and malignant or benign DBSs were identified based on surgical findings, endoscopy findings, or follow-up. The diagnostic accuracy of the two reviewers using MRCP alone and MRCP combined with CT were evaluated. MRCP and CT features of malignant and benign DBSs were compared using multiple logistic regression analysis to identify independent malignant risk factors. RESULTS: MRCP combined with CT examination could improve the diagnostic accuracy, which increased from 70.2% to 81.5% in Doctor A and from 85.1% to 89.3% in Doctor B. The multiple logistic regression model revealed that stricture length [odds ratio (OR) 1.070, P=0.016], angle of the DBS (OR 1.061, P<0.001), double duct sign (OR 4.312, P=0.003) and low density in the arterial phase (OR 0.319, P=0.018) were associated with malignant DBS. A scoring model incorporating these four factors was established; at a threshold value of 1.75, and the sensitivity and specificity for the detection of malignant DBSs were 73.5 and 85.9%, respectively. CONCLUSIONS: Compared to the use of MRCP alone, MRCP combined with contrast-enhanced CT can improve the accuracy of DBS diagnosis. The scoring model accurately predicts malignant DBSs and helps make treatment decisions.

A synthetic MD-2 mimetic peptide attenuates lipopolysaccharide-induced inflammatory responses in vivo and in vitro.

Myeloid differentiation protein-2 (MD-2), a secreted glycoprotein that binds to both lipopolysaccharide (LPS) and toll like receptor 4 (TLR4), contributes to the fine ligand recognition and signaling activation on LPS-induced inflammation. Here we synthesized a novel MD-2 mimetic peptide (MDMP), derived from the putative LPS-binding domain and TLR4-binding domain of MD-2, and found that MDMP dose-dependently bound to LPS and inhibited LPS-activated Limulus amebocyte lysate (LAL). Pretreatment with MDMP dampened LPS-induced inflammatory responses in RAW264.7 cells, including down-regulation of TLR4-MD-2 complex on the cell surface, suppression of LPS binding to the cells, inhibition of mitogen-activated protein kinase (MAPKs) and nuclear factor kappa B (NF-kappaB) activation, reduction of tumor necrosis factor-alpha (TNF-alpha) production. Further, in vivo pretreatment with MDMP markedly protected against LPS-induced acute lung injury and liver injury, as indicated by the notable reduction of lethality, inflammatory responses and TNF-alpha production. These results demonstrate that MDMP attenuates LPS-induced inflammatory responses in vivo and in vitro, and suggests that MDMP may be useful in the treatment of inflammation associated with LPS.

[Construction of human phage antibody library and screening and characterization of phage antibodies against N terminal fragment of human lipopolysaccharide binding protein].

AIM: To construct human phage antibody library and to prepare phage antibodies against N terminal fragment of human lipopolysaccharide binding protein (NH-LBP). METHODS: NH-LBP was expressed in insect cells sf21 cultured with serum-free culture medium (SF-900 II) by BAC-TO-BAC baculovirus expression system. Human antibody library (Fab) was constructed by phage display system (pComb3H/VCSM13) and screened with NH-LBP. RESULTS: NH-LBP was successfully expressed in insect cells sf21 cultured with serum-free culture and effectively purified via affinity chromatography column with TALON resin. About 8 mg NH-LBP was obtained. Human combinatorial phage display antibody library consisting of 5.0 x 10(8) CFU was constructed. After eight rounds of panning with NH-LBP we got 3 phage antibodies with high affinity for NH-LBP (DNA sequences and amino acid sequences of kappa chains of these antibodies were deposited in GenBank (AY337713 and AY337714). CONCLUSION: Phage antibodies against NH-LBP were obtained successfully, which lays the foundation for preparing disulfide stabilized Fv fragments antibody (dsFv) against NH-LBP, understanding the function of LBP in vivo, and prevention and therapy of excessive inflammatory reaction.

[Study on the protective effect of recombinant human N-terminal lipopolysaccharide binding protein in mice challenged with LPS].

OBJECTIVE: To investigate the protective effect of recombinant human N-terminal lipopolysaccharide binding protein in mice challenged with LPS. METHODS: Seventy male Kunming mice were randomly divided into 3 groups, i.e. LPS challenge (Injection of LPS into abdominal cavity, n = 21); tLBP protection (Injection of LPS and tLBP into abdominal cavity, n = 21) and control (Injection of normal saline into abdominal cavity, n = 8) groups. The blood samples and tissue samples of the liver and lungs were harvested on 15 and 30 minutes and 1, 3, 6, 12 and 24 hours after the injection. The serum contents of ALT and TNF-alpha were determined by biochemical velocity analysis and RIA method, respectively. The pathomorphological changes in the liver and pulmonary tissue were examined under light microscope (LM). The mortality rate of ten mice each was observed within 24 hours after the injection of tLBP + 400 ng LPS or 400ng LPS. RESULTS: The ALT content of tLBP group reached the peak level at 12 post-injection hour (PIH) (41.00 +/- 4.58), but it was significantly lower than that in LPS group in which it peaked at 6PIH (99.50 +/- 62.63) (P < 0.01). The TNF-alpha content in tLBP and LPS group was lower than that in LPS group, and both reached the peak level at 3 PIH (35.96 +/- 7.33). Compared with those in LPS, injury to hepatocytes in tLBP group was obviously milder without scattered necrosis. The pulmonary congestion in tLBP group was abated, and the inflammatory exudation in the alveoli was evidently less than that in LPS group. There were 9 out of 10 mice died in the LPS challenge group, while only 3 out of 10 mice died during 24 hours after LPS injection in tLBP protection group. CONCLUSION: Preliminary results indicated that recombinant human tLBP might possess biological activity with a potential protection effect in LPS challenged mice.

[The purification and isolation of a new type of rabbit-origin lipopolysaccharide binding protein and the study of its biological function in vitro].

OBJECTIVE: To isolate and purify a new type of lipopolysaccharide binding protein (LBP) from burn rabbit serum, and to investigate its biological functions. METHODS: Rabbits subjected to burn injury and endotoxemia were employed. The serum from the rabbits was purified by two-steps of ion-exchange chromatography (Bio-Rex 70 Resin, Mono-Q) and gel chromatography. Furthermore, the serum was identified by flow cytometry analysis, agglomeration test with sheep erythrocyte, and amino end amino acid residue sequencing. The obtained protein was applied to cultured human monocytes (U937), and the cytokine secretion such as TNFalpha from the U937 was observed. RESULTS: The molecular weight of the harvested protein was 48 kDa, and the 10 amino acid sequence at N end was arranged as GSQGTFTSEE, which was different to the amino acid sequence in NCBI protein bank and was so named P48. P48 possessed similar function to that of LBP and could promote the binding of LPS in a very low concentration with peripheral blood monocytes (PBMC), and also promote the TNFalpha secretion from U937. CONCLUSION: P48, a new type of LBP, could be isolated and purified from the burn rabbit serum. P48 possessed similar biological activities to that of LBP and could promote the process of inflammatory reaction.