[Expression and significance of c-fos in resistant cell line TU177/VCR of larynx squamous cell carcinoma].

Objective: To explore the effect of c-fos on multidrug resistance of laryngeal cancer TU177 cells. Method: Increasing drug concentration gradient is adopted to establish the stability of the laryngeal cancer drug resistance in cell line; RT-PCR and Western blot were used to detect difference of the c-fos between TU177 and TU177/VCR cells; plasmids with human c-fos knockdown or over expression were transfected into TU177/VCR and TU177 cells respectively, and the effects of different treatment on cell proliferation were investigated with MTT. Results: The drug resistance of TU177/VCR cells was 26.25-fold in vincristine (VCR), 7.33-fold in Paclitaxel (TAX), 2.41 in cisplatin (DDP), and 5.50 in 5-fluorouracil (5-FU), comparing with TU177( P<0.05). The TU177/VCR cells had significantly higher c-fos expression compared to TU177 cells( P<0.05). The results showed that the IC(50) values of 5-FU for the NC group and c-fos shRNA group were (306.2±6.3)µmol/L and (81.3±3.9)µmol/L, respectively, which was decreased by 73% in the c-fos shRNA group compared to that in the NC group (P<0.05). Similarly, the results showed that the IC(50) values for 5-FU were (55.3±9.4) µmol/L in NC group and (288.1±7.3)µmol/L in c-fos WT group, which was increased 5.21-fold in c-fos WT cells. Conclusion: C-fos plays important role in multidrug resistance of larynx cancer cell TU177/VCR, and might become a new molecular target for laryngeal cancer treatment.

[FAT1 inhibits cell proliferation of esophageal squamous cell carcinoma through regulating the expression of CDK4/CDK6/CCND1 complex].

Objective: To explore the expression of FAT1 in esophageal squamous cell carcinoma (ESCC) tissues, and its effect on cell proliferation. Methods: The expression levels of FAT1 protein in human ESCC tissues and matched adjacent normal tissues were determined by immunohistochemistry (IHC). Lentivirus based knockdown of FAT1 was carried out in YSE2 and Colo680N cell lines and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assays was performed to examine the effect of FAT1 on the proliferation of these ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle and apoptosis. The expression levels of cell cycle markers in FAT1 knock out ESCC cell lines were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR) and Western blot. Results: The relative expression of FAT1 in ESCC tissues was 66.97±21.53, significantly lower than 78.13±16.76 of adjacent normal tissues(P<0.05). Knockdown of FAT1 promoted cell proliferation and colony formation. In YSE2 cell, the division time in negative control (NC) group was (1 570±51) min, significantly longer than (1 356±31) min in shFAT1 group. In Colo680N cell, division time in NC group was (1 532±53) min, significantly longer than (1 290±30) min in shFAT1 group (P<0.05). Knockdown of FAT1 promoted G1-to S-phase transition and resulted in the upregulation of CDK4/CDK6/CCND1. Conclusion: FAT1 inhibits the proliferation and G1-to S-phase transition of ESCC cells through regulating the protein expression of CDK4/CDK6/CCND1 complex.

An experimental study: quantitatively evaluating the change of the content of collagen fibres in penis with two-dimensional ShearWave™ Elastography.

The purpose of this study was to explore the value of two-dimensional ShearWave™ Elastography (2D-SWE) on quantitatively evaluating the change of the content of collagen fibres in penis. Twenty male Sprague Dawley rats were divided into the pre-sexual maturity group (Group 1) and the sexual decline group (Group 2) according to age. The ultrafast ultrasound device Aixplorer® (SuperSonic Imagine, Aix-en-Provence, France) was used for 2D-SWE imaging of penis, and the measurement index was shear wave stiffness (SWS). The immunohistochemistry was used to analyse the content of collagen fibres in penis, and the measurement index was positive area percentage (PAP). The differences of SWS between the two groups and PAP between the two groups were analysed. SWS of Group 1 and Group 2 was 10.18 ± 1.09 and 8.02 ± 1.34 kPa, and SWS of Group 2 was significantly lower than Group 1 (p < .01). PAP of Group 1 and Group 2 was 4.83 ± 3.61% and 16.41 ± 10.02%, and PAP of Group 2 was significantly higher than Group 1 (p < .01). Our results indicate that when the content of collagen fibres changes, SWS of penis measured with 2D-SWE would change significantly as well. Two-dimensional SWE can be used to quantitatively evaluate the change of the content of collagen fibres in penis.

An experimental study: evaluating the tissue structure of penis with 2D-ShearWave™ Elastography.

The aim of this study was to investigate the feasibility of two-dimensional-ShearWave™ Elastography (2D-SWE) on evaluating the change of tissue structure of penis. Twenty healthy male Sprague Dawley rats were divided into penis-developed group (PDG, 52 weeks) and penis-underdeveloped group (PUDG, 5 weeks). The ultrafast ultrasound device-Aixplorer® (SuperSonic Imagine) was used for 2D-SWE imaging of the penis, the measurement index was shear wave stiffness (SWS, kPa). All rat penises were cut off immediately after ultrasonic examination. After paraffin embedding, slicing and hematoxylin-eosin staining, the tissue structure of the penis was observed under light microscope. SWS of all rat penises were measured successfully. The results showed that SWS of PDG was significantly lower than PUDG (P=0.008). At the same time, the pathological results found that there were significant differences in the tissue structures (sinusoids, smooth muscle cells and fibrocytes) of the penises between the two groups. These results suggest that there are significant differences in SWS between different tissue structures of penis. 2D-SWE is expected to be used on the etiological diagnosis of erectile dysfunction by serving as a new noninvasive method of evaluating the change of tissue structure of penis.

A new method of measuring the stiffness of corpus cavernosum penis with ShearWave™ Elastography.

OBJECTIVE: To evaluate the feasibility of measuring the stiffness of corpus cavernosum penis (CCP) with ShearWave™ Elastography (SWE; SuperSonic Imagine, Aix-en-Provence, France). METHODS: 40 healthy volunteers with ages ranging from 19 to 81 years (mean, 36 years; standard deviation, 17 years) were selected in this study. The ultrafast ultrasound device Aixplorer(®) (SuperSonic Imagine) was used for the research and the probe selected was SuperLinear™ SL15-4 (SuperSonic Imagine). The shear wave stiffness (SWS) of CCP was measured using SWE images. The measurement indexes of SWS included (1) SWS of CCP measured in the transverse section (SWS-T), (2) SWS of CCP measured in the longitudinal section (SWS-L) and (3) mean of SWS-T and SWS-L (SWS-M). The interval between hormone test and SWE examination of each subject was less than 7 days. The paired t-test was used to analyse the differences between SWS-T and SWS-L. The Pearson correlation was used to analyse the correlation of SWS of CCP with age as well as with sex hormone levels. RESULTS: There was no significant difference between SWS-T and SWS-L (p > 0.05). SWS (SWS-T, SWS-L, SWS-M) was negatively correlated with age and oestradiol value, and SWS (SWS-T, SWS-L, SWS-M) was positively correlated with testosterone value. CONCLUSION: SWE could serve as a new non-invasive method of evaluating the stiffness of CCP. ADVANCES IN KNOWLEDGE: It is the first time that we have discussed the feasibility of measuring the stiffness of CCP with SWE and analysed the correlation of SWS of CCP with age as well as with sex hormone levels.

An assessment of teicoplanin use and monitoring serum levels in a Chinese teaching hospital.

OBJECTIVE: To validate a high performance liquid chromatography (HPLC) method for serum teicoplanin measurement and use the method for clinical monitoring of teicoplanin levels to analyze the clinical application of teicoplanin. METHODS: 55 patient profiles were collected and analyzed for the clinical teicoplanin application. 10 critically ill patients of the 55 cases were monitored for teicoplanin trough concentration using the HPLC method. RESULTS: The modified HPLC method exhibited excellent linearity, with correlation coefficient r = 0.9995. The intra-day and inter-day coefficients of variation were less than 10%. The lower limit of detection of teicoplanin was 5.63 mg/l. The recovery of teicoplanin was above 90%. Of the 55 patients in this study, there were 42 patients without load-dosing. There were only 29 patients treated with teicoplanin documented Gram-positive infections by etiological diagnoses. In the 10 patients with teicoplanin serum trough concentration monitoring, all cases received a loading dose of 400 mg every 12 h for 3 doses, and the mean trough concentration of teicoplanin was 10.82 ± 4.51 mg/l. The mean trough levels were 13.04 ± 6.23 mg/l in 4 patients with microbiological eradication and improvement of symptoms of diseases and 9.34 ± 2.61 mg/l in 6 patients with persistence of previous clinical infectious symptoms, respectively. CONCLUSION: The modified HPLC method is robust, highly reproducible and suited to monitor the concentration of teicoplanin. In critically ill Chinese patients, we should consider more appropriate loading doses and evaluate the relationship between teicoplanin trough concentration and the efficacy using microbiological and clinical parameters.

[Photodegradation assay method of nifedipine and its application to studies on percutaneous absorption].

A new method of analysis for low concentrations nifedipine was developed according to the principle of a photodegradation analytical method that has been reported by the authors previously, and was used to study percutaneous absorption. The absorbance of sample solution was measured before and after light irradiation at 237 nm for 2 h. In this method, calibration graph was linear in the range of 1-20 micrograms/ml for delta A237. The average recovery for nifedipine was 98.80%. No interference from propylene glycol, azone, m-nifedipine, nitrendipine, verapamil and propranolol was observed. It is shown that azone can promote markedly percutaneous absorption of nifedipine.

[Determination of m-nifedipine and its pharmacokinetic study in rabbits by high-pressure liquid chromatography].

A high-pressure liquid chromatographic method was developed for determination of m-nifedipine in plasma using a chemical bonded C-18 phase column (YWG-C18 10 microns, made in China) with nitrendipine as internal standard. To increase life of the YWG-C18 column a mixture of methanol and 5 mmol.L-1 phosphate buffer (70:30 vol/vol) was selected as mobile phase with a flow rate of 0.8 ml.min-1. The method was sensitive to m-nifedipine 3 ng.ml-1 plasma and the standard curve was linear from 10 to 1000 ng.ml-1 with correlation coefficient of 0.99. The within-day and day-to-day precisions (CV) of this method were 4.5% and 7.0%, respectively, with recoveries of 95-102% (10-1000 ng.ml-1). There was no interference with nifedipine, amiodarone, propranol, and verapamil. A pharmacokinetic study on m-nifedipine was carried out in 8 rabbits. A better computer fitted to a two-compartment model was observed using 3P87 program. The parameters obtained were as follow: Vc 6.3 L.kg-1, Cl 0.021 L.kg-1.min-1, T1/2 alpha 30 min, T1/2 beta 230 min, AUC 102 micrograms.min.ml-1.