Objective The present study was to elucidate whether there was a local renin-angiotensin-aldosterone system (RAAS)in rat hepatic stellate cells(HSC)and the immortalized rat HSC line—HSC-T6. Methods HSC was isolated by perfusion of collagenase and pronase to liver. The renin activity, angiotensin Ⅱ(Ang Ⅱ) and aldosterone(ALD) concentration in the cells were assayed by radio-immunoassay. Angiotensin converting enzyme(ACE) activity was detected by ultra-violet spectrophotography. The expression of angiotensin Ⅱ type 1 receptor(AT1R) was determined by immunohistochemistry. RT-PCR method was used to assay the mRNA expression of renin, angiotensinogen, ACE, AT1R and CYPⅡB2(the critical enzyme of aldosterone synthesis). Results Ang Ⅱ, ALD, renin and ACE activity could be detected in HSC and HSC-T6. Immunohistochemistry showed that both of them could express AT1R. mRNAs of renin, ACE, AT1R and CYPⅡB2 were also detected in both of them. Conclusions Both rat HSC and the immortalized rat HSC line HSC-T6 possessa a local RAAS.

AIM　Diclofenac potassium sustained-release tablets were developed as a gel matrix for using twice daily and evaluated in vitro release characteristics. METHODS　The formulations were screened according to the designed in vitro release rate by the use of HPMC as the gel matrix and the hydrophobic retarding agent to modify the release. RESULTS　The release was properly characterized by the diffusion mechanism, influenced by pH of the media, slightly affected by the basket rotating speed, without the influences from the pressure exerted during the tabletting procedure. CONCLUSION　The selected formulation of diclofenac potassium sustained-release tablets could ensure the desired in vitro release rate. In addition, it proved a good manufacturing reproducibility.

Humans ; Integrative Medicine ; Lupus Vasculitis, Central Nervous System ; therapy

Objective To explore whether CCL18 is involved in regulating the expression of miRNAs in breast cancer.Methods The expression profile of miRNAs in the breast cancer cell following CCL18 treatment was determined by miRNAs microarray analysis.Then we performed QRT-PCR and Luciferase Reporter Assay to validate the results from the miRNAs microassay.We used transient transfection to change the expression of miR98 and c-myc in breast cancer cells.We then used QRT-PCR and Western blot to analyze the mechanism by which CCL18 downregulates the expression of miR98 in breast cancer cells.Results miRNAs microarray analysis showed that cells treated with CCL18 differentially expressed 20 miRNAs genes compared with those in the control group. Our QRT-PCR and Luciferase Reporter Assay confirmed the result.The mRNA and protein expressions of C-myc and lin28 were increased after CCL18 stimulation in breast cancer cells.Transfection with c-myc siRNAs rescued the increase of lin28 and loss of miR98 expression caused by CCL18 stimulation.Our results also showed that CCL18 could upregulate the expression of N-Ras at post-transcription level.Conclusion CCL18 downregulates the expression of miR98 via N-Ras/c-myc/lin28 pathway.The downregulated miR98 increases the expression of N-Ras after transfection,which further activates c-myc/lin28 pathway and forms a positive feedback loop.

Objective To compare the effects of different ischemic preconditioning (IP) protocols on the viability of primary cultured sinoatrial node cells from neonatal rats for investigating the protective effect of IP. Methods The cells were randomized into eleven groups: control, ischemia/reperfusion (I/R), IP1 (preconditioned with ischemia/reperfusion for 5 min), IP2 (preconditioned with 2 cycles of ischemia/ reperfusion for 5 min), IP3 (preconditioned with 3 cycles of ischemia/reperfusion for 5 min), IP4 (preconditioned with ischemia/reperfusion for 10 min), IP5 (preconditioned with 2 cycles of ischemia/reperfusion for 10 min), IP6 (preconditioned with 3 cycles of ischemia/reperfusion for 10 min), IP7 (preconditioned with ischemia/reperfusion for 20 min), IP8 (preconditioned with 2 cycles of ischemia/reperfusion for 20 min), and IP9 (preconditioned with 3 cycles of ischemia/reperfusion for 20 min). PI positive staining rate and changes of D 490 after 3 h ischemia/4 h reperfusion were determined by PI staining and MTT chromatometry. Results ① D 490 value in sinoatrial node cells in each experimental group was significantly lower than that in the control group, but that in IP2, IP3, IP4, IP5, and IP7 groups was significantly higher than that in I/R group (P

ObjectiveTo investigate the effect of volume embolization ratio(VER) in evaluating the embolization efficacy of aneurysms.MethodsFifty-three patients who received intracranial aneurysm embolization surgery were all treated with detachable coils endovascular embolization,While fifty-seven intracranial aneurysms were treated.The VER was calculated and the relationships between the VER values and the size,the neck,embolization degree of intracranial aneurysm were analyzed.ResultsThe VER of small,medium,large,and giant aneurysms were (40.8 ± 26.5 )％,( 18.6 ± 16.1 )％,(2.3 ± 2.1 )％,(0.4 ±0.1 )％,respectively,which had statistical significance (F =7.091,P ＜ 0.01 ).The VER of wide-necked and narrow-necked aneurysms were (27.5 ± 23.1 )％,(29.4 ± 26.6)％,which had no statistical significance(P ＞0.05 ).The VER of complete embolization,neck residual and partial embolization were (41.8 ± 29.3 )％,(31.4 ± 21.2)％,( 12.3 ± 15.1 )％,respectively,which had statistical differences (F =7.970,P ＜ 0.01 ).ConclusionsThe VER is an objective index to evaluate the embolization degree of aneurysms.It has certain prediction significance for the efficacy judgement of embolization of aneurysms.

Objective To evaluate the efficacy and safety of endoscopic mucosal resection (EMR) for rectal carcinoid tumors. Methods From January 2006 to January 2009, EMR was performed in 28 patients with rectal carcinoids, who were followed up to evaluate the therapeutic effect and safety. Results Tumor diameters varied from 0.4 cm to 1. 2 cm (mean 0.7± 0. 2 cm). Negative resection margin was a-chieved in 26 cases (92. 9% ), tumor margin within 0. 1 cm of resection margin in 1 (7. 1% ) , and two margins coincided in 1 patient (7. 1% ). Hemostasis was performed with metal clips in 14 patients (50% ) and argon plasma coagulation (APC) in 9 (32. 1% ). Except for rectal bleeding in 1 patient (3. 6% ) , no other complications were observed. There was no recurrence in any patients during a follow-up of 6-36 months. Conclusion EMR is a useful and safe method for treatment of small rectal carcinoid tumor which does not cross submucosal layer.

Aim To establish a LC-MS/MS method for the determination of pirfenidone(BT)and its major metabolite 5-carboxy-pirfenidone(SBT)in human plasma.Methods Human plasma samples containing BT and SBT,as well as their corresponding deuterium-labeled internal standards pirfenidone-d5(dBT)and 5-carboxy-pirfenidone-d5(dSBT),were precipitated using methanol.Chromatographic separation was carried out on an Agilent ZORBAX SB C18(3.0 mm×100 mm,3.5 μm)column with the mobile phase of water(0.5％formic acid)and acetonitrile(50/50).The detection of analytes was performed on a tandem mass system equipped with an electrospray ionization source in positive ion mode using multiple-reaction monitoring.The MS/MS ion transitions monitored were m/z 185.958→77.1 for BT,m/z 215.944→77.0 for SBT,m/z 190.965→81.1 for dBT and m/z 220.948→99.1 for dSBT.Results There was no remarkable interference in blank solvent,plasma,and there was no mutual interference between analytes or internal standards.The proposed method showed good linearity over the concentration range of 0.020 59～25.14 mg·L-1 for BT and 0.016 73～20.42 mg·L-1 for SBT.The intra-batch and inter-batch precision and accuracy were proved to be acceptable.Human samples kept stable after 4 h at room temperature,the three freeze-thaw cycles and 10,29 and 52 days at-70 ℃,and the processed samples remained stable after 24 h in the autosampler.The average extraction recovery and matrix effect were precise,reproducible and acceptable.Conclusion Our current LC-MS/MS method is proved to be sensitive,accurate and convenient,and could be suitable for the clinical pharmacokinetic studies of BT-related preparations.

Background In the past few decades,the balance of Th1/Th2 is often used to explain the immune mechanisms of fungal infection and fungal disease.More recently,a novel subset of CD4+ effector Th cells has been found to participate in anti-fungal infection response.However,whether Th17 is involved in the immune response in fungal keratitis is unclear up to now.Objective Present study was to investigate the expression change of Th17 type cytokine and its specific transcription factor,retinoid-related orphan nuclear receptor gamma t (RORγt),in the cornea of Fusarium solani keratitis.Methods Ninety-six clean BALB/c mice were divided into Fusarium solani keratitis model group and control group by randomized digital table.Fusarium solani keratitis models were established by epikeratophakia-assisted corneal epithelial erasion and interlayerly injection of 5 μl (1 × 106 CFU/ml) Fusarium solani solution in the right eyes,and the equal volume of PBS was injected in the same way in the control group.10％ KOH wet film was used to examine the fungal hyphea and funga strain was identified by inoculation.The corneas were examined under the slit lamp microscope 1 day,3,5,7 days after modeling and the inflammatory response was scored based on the criteria of Wu and Hu.The histopathological examination of corneas was performed in the time points above.Real time fluorescence quantitative PCR was used to detect the expression levels of interleukin-17 (IL-17) mRNA and RORγt mRNA in the corneas.The expression of IL-17 protein in the corneas was detected by ELISA.The use and raise of the mice followed the Statement of Association for Research in Vision and Ophthalmology.Results The inflammatory scores were 3.2±0.8,6.6± 1.1,9.4± 1.1 and 6.8±0.8 in 1 day,3,5,7 days after modeling,showing a significant difference among them (F =89.786,P =0.010).The inflammatory scores were higher in the third and seventh day than that in the first day (P＜0.05),but they were significantly lower than that in the fifth day (P＜0.05).The infiltration of inflammatory cells showed a coincident tendency with the score.The expressing levels of IL-17 mRNA (2-ΔΔCt) in the corneas were 4.12±0.73,20.72±1.81 and 14.16±1.88 in 3,5,7 days after modeling,with statistically significant differences in comparison with those in the control group (P＜0.01),and the expression level was significantly higher in the fifth day than those in the first,third and seventh day in the model group(P＜0.01).The expression levels of IL-17 protein (ng/g) were significantly increased 1 day,3,5,7 days in the model group compared with the control group (P＜0.01).A similar change was found in the expression of RORγt mRNA to that of IL-17 mRNA.Conclusions Expressions of IL-17 and its transcription factor RORγt upregulate in the fungal keratitis and has an association with inflammatory degree,which suggests that Th17 subset may play an important role in the immune responses of fungal keratitis.

Objective To analyze the views and perceptions of practicing physicians on the practicing environment in the province,in order to provide policy reference for improvement.Methods By means of equi-probability and multi-stage sampling, 3 570 valid questionnaires were collected and analyzed from site survey which consist of the subj ective evaluation,and measures to identify and prevent medical professional risk.Results Overall assessment of the practice environment of physicians in Shanxi province is low,as those holding the environment asniceandacceptableaccounting for 4.8% and 21.7%,only 47.0% of physicians in tertiary hospitals hold the environment as very bad,while only 6.1% of those in level-1 hospitals share this view.92.4% of the physicians surveyed hold compliance of laws and regulations and technical specifications as key to prevention of medical dispute,yet most of them do not expect the media to be obj ective,fair and accurate in their coverage of events.Conclusions It is recommended to strengthen communication with the media for obj ective coverage of the limits of medicine,and explore the mechanism to take care of medical disputes by physicians,and improve their medical risk control capabilities,for a better practicing environment.