The mechanisms of paeonol reversing multidrug resistance in ovarian cancer SKOV3/DDP cells / 药学学报

The purpose of this research is to investigate the effects and mechanisms of paeonol (PL), a phenolic compound found in many traditional Chinese formulations, on reversing drug resistance in the ovarian cancer resistant SKOV3/DDP cells. The results showed that PL had significant drug-resistant reversal effect on SKOV3/DDP cells. Flow cytometry showed that PL could inhibit P-glycoprotein (P-gp) function in a concentration-dependent manner. Fluorescent quantitative PCR and cell immunofluorescence techniques were used to detect mechanisms of action. Results revealed that both the inhibitory effect on MDR1/P-gp and metadherin (MTDH) expression and the induction effect on phosphatase and tensin homolog (PTEN), by 15, 30, and 60 μmol·L-1 PL, were increased with increased concentrations of PL (P P -1 PL treated group; however, the inhibition or induction effect on MTDH or PTEN protein were only comparable to the 15 μmol·L-1 PL treated group. The present study shows that the effect of PL on SKOV3/DDP cells may be related to the inhibition of P-gp function and expression, the inhibition of MDR1, MTDH expression, and the induction of PTEN expression, all which can provide a theoretical foundation for PL as a drug resistance reversal agent on the treatment of ovarian cancer chemotherapy resistance.

Guizhi Fuling Wan reverses drug resistance by regulating PTEN and MTDH in ovarian cancer SKOV3/DDP Cells / 中国药理学与毒理学杂志

OBJECTIVE To detect the reversal effect of Guizhi Fuling Wan on cisplatin-resistant ovarian cancer SKOV3/DDP cells and its relationship with protein expression of phosphatase and tensin homolog (PTEN) and metadherin (MTDH). METHODS Guizhi Fuling Wan (GFW) concentrated solution was prepared according to the Chinese Pharmacopoeia 2015 edition, Wistar rats were given GFW viagavage at 4 g·kg-1·d-1,8 g·kg-1·d-1,16 g·kg-1·d-1,or given saline as blank control for 5 days.Blood samples were taken and the corresponding drug-containing low-dose sera, medium-dose sear, high-dose sera and blank sera were prepared.The XCELLigence RTCA S16 real-time label-free cell analyzer was used to detect the reversal effect by the sera combined with cisplatin or paclitaxel in SKOV3/DDP cells. Annexin V-FITC and PI double-staining were used to detect the apoptosis-inducing effect of the sera in the cells. RT-qPCR and western blot were used to detect the mRNA and protein expression of PTEN and MTDH after the cells treated with the sera. RESULTS The inhibition rate of low-dose sera against SKOV3/DDP cells was less than 5%.After the low-dose sera combined with cisplatin or pacli-taxel, the IC50 of SKOV3/DDP cells against cisplatin and paclitaxel decreased by 3.01 and 1.79-fold, respectively.The total apoptosis rates induced by the low-dose sera,medium-dose sear,high-dose sera and blank sera in SKOV3/DDP cells were 11.08±0.13,19.42±0.30,24.23±0.31,and 3.21±0.24,respec-tively; there was a significant difference between the groups (P<0.01). RT-qPCR results showed that, compared with the blank serum, the sera can up-regulate the expression of PTEN mRNA and down-regulate the expression of MTDH mRNA in a dose-dependent manner. Western blot results showed that the induction effect to PTEN protein and the inhibition effect to MTDH protein by the sera were gradually enhanced with thesera dose increasement. CONCLUSION The resistance reversal effect of Guizhi Fuling Wan on ovarian cancer SKOV3/DDP cells may be related to the inhibition of MTDH, up-regulation of PTEN and induction of apoptosis, providing with an experiment basis for the applica-tion of Guizhi Fuling Wan as a reversal agent for chemotherapy resistance of ovarian cancer.

Incidence of pleural effusion in patients with pulmonary embolism / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>No data on the incidence of pleural effusion (PE) in Chinese patients with pulmonary embolism are available to date. The aim of the current study was to investigate the frequency of PE in a Chinese population of patients with pulmonary embolism.</p><p><b>METHODS</b>This was a retrospective observational single-center study. All data of computed tomography pulmonary angiography (CTPA) performed over 6-year period on adult patients with clinically suspected pulmonary embolism were analyzed.</p><p><b>RESULTS</b>From January 2008 until December 2013, PE was identified in 423 of 3141 patients (13.5%) with clinically suspected pulmonary embolism who underwent CTPA. The incidence of PE in patients with pulmonary embolism (19.9%) was significantly higher than in those without embolism (9.4%) (P < 0.001). Majority of PEs in pulmonary embolism patients were small to moderate and were unilateral. The locations of emboli and the numbers of arteries involved, CT pulmonary obstruction index, and parenchymal abnormalities at CT were not associated with the development of PE.</p><p><b>CONCLUSIONS</b>PEs are present in about one fifth of a Chinese population of patients with pulmonary embolism, which are usually small, unilateral, and unsuitable for diagnostic thoracentesis.</p>

Predicting value of various clinical probability scores for diagnosis of lower limb deep venous thrombosis in Chinese patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the predicting value of Wells, Kahn, St. André and Constans scores for the diagnosis of deep venous thrombosis in Chinese patients.</p><p><b>METHODS</b>A total of 274 patients suspected with deep venous thrombosis was prospectively blinded evaluated with the four clinical-score systems. Sensitivity, specificity, positive predictive value, negative predictive value and receiver operation curves were calculated for four clinical scores according sonography results.</p><p><b>RESULTS</b>Sonography evidenced deep venous thrombosis in 88 out of 274 patients. The sensitivity, specificity, positive predictive value and negative predictive value was 77.3%, 65.6%, 51.5% and 85.9%, respectively, for Wells score; 58%, 55.9%, 38.3% and 73.8%, respectively, for Kahn score; 64.8%, 55.4%, 40.7% and 76.9%, respectively, for St. André score and 86.4%, 37.6%, 39.6% and 85.4%, respectively, for Constans score. ROC was 0.761 for Wells score, which was similar as that of Constans score (0.759), then followed by St. André score (0.627) and Kahn score (0.591).</p><p><b>CONCLUSION</b>Our results showed that Wells score and Constans score are superior to Kahn score or St. André score for diagnosing patients with suspected deep venous thrombosis in terms of sensitivity, negative prediction value and ROC values.</p>

SKOV-3 cell imaging by paramagnetic particles labeled with hairpin cell-penetrating peptides / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The hairpin cell-penetrating peptides (hCPPs) demonstrate an interesting characteristic of conditioned activation by molecules. We hypothesized that hCPPs have the potential to selectively deliver a paramagnetic gadolinium probe into the matrix metalloproteinase 2 (MMP-2) positive human ovary adenocarcinoma cell lines, SKOV-3.</p><p><b>METHODS</b>hCPPs were synthesized and labeled with 1,4,7,10-tetraazacyclododecane-N,N',N'',N''' tetraacetic acid gadolinium (III) (Gd-DOTA) and fluorescein isothiocyanate (FITC) by f-moc strategy using a standard solid phase peptide synthesis protocol. MMP-2 expression and activity were demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Internalization and location of hCPPs in SKOV-3 cells were observed by fluorescein imaging and flow cytometery. Selective delivery of Gd-DOTA in SKOV-3 cells was observed by magnetic resonance imaging (MRI) and transmission electron microscopy (TEM).</p><p><b>RESULTS</b>The uptake of hCPPs by SKOV-3 cells depended on the activity of MMP-2. T1WI signals of SKOV-3 cells treated with Gd-DOTA-hCPPs suggested the uptake of Gd-DOTA-hCPPs increased in a time- (r = 0.990, P < 0.01) and concentration-dependent manner (r = 0.964, P < 0.001), but was inhibited by a MMP-2 inhibitor. Electron-dense particles observed in the cytoplasm and nucleus by transmission electron microscopy proved the intracellular penetration of gadolinium.</p><p><b>CONCLUSIONS</b>hCPPs can be used as an effective vector for an MRI molecular probe to assess the activity of MMP-2.</p>

Value of combined wells score and D-dimer test on diagnosing patients with deep venous thrombosis / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the value of Wells score or/and D-dimer test on diagnosing or excluding deep venous thrombosis (DVT).</p><p><b>METHODS</b>Patients with suspected DVT were retrospectively analyzed. All patients underwent clinical assessment, D-dimer assay and bilateral lower extremity compression sonography within 48 hours of admission. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for diagnosing DVT by Wells score, D-dimer test, and combined Wells score and D-dimer were compared.</p><p><b>RESULTS</b>A total of 274 patients were analyzed. If low probability was defined as negative and moderate and high probabilities were defined as positive, the sensitivity, specificity, PPV and NPV of the Wells score were 78.4%, 66.1%, 52.3% and 86.6%, respectively. At a cut-off of 500 microg/L, the sensitivity, specificity, PPV and NPV of D-dimer test were 73.9%, 66.1%, 50.8% and 84.2%, respectively. If low probability and D-dimer < 500 microg/L were defined as negative, moderate and high probabilities and D-dimer > or = 500 microg/L were defined as positive, the sensitivity, specificity, PPV and NPV of the combined Wells score and D-dimer test were 88.3%, 76.8%, 67.1% and 92.5%, respectively.</p><p><b>CONCLUSION</b>For clinical suspected DVT patients, DVT diagnosis could be reliably obtained by combined Wells score and D-dimer test.</p>

Study on the risk factors regarding deep venous thrombosis in acute pulmonary thromboembolism / 中华流行病学杂志

<p><b>OBJECTIVE</b>To identify the risk factors of deep venous thrombosis(DVT) in patients with acute pulmonary thromboembolism (PTE).</p><p><b>METHODS</b>Post hoc analysis on data from a prospective multicenter through a randomized control trial on thrombolysis and anticoagulant among 454 patients with acute symptomatic PTE from June 2002 to December 2004. All patients were confirmed PTE by CT pulmonary angiography (CTPA) and/or ventilation perfusion scan or by magnetic resonance pulmonary angiography. Data of PTE patients with DVT was compared with those without DVT to identify the DVT risk factors. The effect of other variable on DVT was assessed by logistic regression analysis.</p><p><b>RESULTS</b>When comparing the age, sex body weight index and underlying diseases including hypertension, coronary heart disease, chronic obstructive pulmonary disease, cerebral accident and malignant tumor between groups of DVT and no DVT,data showed that there was no statistically significant difference except for diabetes (chi2 = 4.481, P = 0.034) among them.However,results from multi-analysis showed that edema of lower-limb [odd ratio (OR), 2.255; 95% CI, 1.493 to 3.408], phlebitis (OR, 2.380; 95% CI, 1.426 to 3.973) and the entire swollen (calf swelling > or =1 cm) larger than asymptomatic side (OR, 3.834; 95% CI, 2.561 to 5.738) were independent risk factors for DVT.</p><p><b>CONCLUSION</b>Edema of lower-limb, phlebitis and calf swelling > or =1 cm seemed to be risk factors for DVT.</p>

A Meta-Analysis of the Accuracy of Prostate Cancer Studies Which Use Magnetic Resonance Spectroscopy as a Diagnostic Tool

OBJECTIVE: We aimed to do a meta-analysis of the existing literature to assess the accuracy of prostate cancer studies which use magnetic resonance spectroscopy (MRS) as a diagnostic tool. MATERIALS AND METHODS: Prospectively, independent, blind studies were selected from the Cochrane library, Pubmed, and other network databases. The criteria for inclusion and exclusion in this study referenced the criteria of diagnostic research published by the Cochrane center. The statistical analysis was adopted by using Meta-Test version 6.0. Using the homogeneity test, a statistical effect model was chosen to calculate different pooled weighted values of sensitivity, specificity, and the corresponding 95% confidence intervals (95% CI). The summary receiver operating characteristic (SROC) curves method was used to assess the results. RESULTS: We chose two cut-off values (0.75 and 0.86) as the diagnostic criteria for discriminating between benign and malignant. In the first diagnostic criterion, the pooled weighted sensitivity, specificity, and corresponding 95% CI (expressed as area under curve [AUC]) were 0.82 (0.73, 0.89), 0.68 (0.58, 0.76), and 83.4% (74.97, 91.83). In the second criterion, the pooled weighted sensitivity, specificity, and corresponding 95% CI were 0.64 (0.55, 0.72), 0.86 (0.79, 0.91) and 82.7% (68.73, 96.68). CONCLUSION: As a new method in the diagnostic of prostate cancer, MRS has a better applied value compared to other common modalities. Ultimately, large scale RCT (randomized controlled trial) randomized controlled trial studies are necessary to assess its clinical value.

Intercellular imaging by a polyarginine derived cell penetrating peptide labeled magnetic resonance contrast agent, diethylenetriamine pentaacetic acid gadolinium / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro.</p><p><b>METHODS</b>Fluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT).</p><p><b>RESULTS</b>The molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group.</p><p><b>CONCLUSIONS</b>The newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.</p>