Objective To investigate the penetrative ability of amlodipine gels and to evaluate their effect on the survival of rat dorsal ischemic random skin flaps.Methods 0.5 ％,1.0 ％,1.5 ％,2.0 ％,and 2.5 ％ amlodipine gels were made.The accumulative penetrative quantities of amlodipine through rat skin were assayed with a modified Franz's diffusion cell in vitro.Pure gel without amlodipine,0.5 ％ and 1.5 ％ amlodipine gel were respectively applied on rat random ischemic skin flap once a day for 7 days.The viable area was measured on the seventh postoperative day and the quantities of amlodipine within skin flap were also detected at 2 and 6 hours after application of amlodipine gel.Results The accumulative penetrative quantities of amlodipine increased in time-and concentration-dependent manner (P＜0.05).Accumulative quantities of 0.5 ％ and 1.0 ％ amlodipine gel were lower than those of 1.5 ％,2.0 ％,and 2.5％ gel,respectively (P＜0.05).The quantities of amlodipine within flap tissue in 1.5 ％ amlodipine gel was higher than that of 0.5 ％ amlodipine gel (P＜0.05).The survival area of flap in 0.5 ％ amlodipine gel group (391.4±65.4) mm2 was higher than those of the pure percutaneous gel group (192.9±56.8) mm2 and the control group (191.0±50.2) mm2(P＜0.05),but no significant difference was seen between 1.5 ％ amlodipine gel group (265.7+88.3) mm2 and control group (P＞0.05).Conclusions Amlodipine could penetrate into skin tissues.0.5 ％ amlodipine gel could significantly increase survival area of ischemic random skin flap.

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Objective To investigate the influence of preoperative chemotherapy of S-1 or capecitabine on the serum CK20mRNA expres-sion in peripheral serum in patients with colorectal cancer.Methods Eighty -seven patients undergoing colorectal cancer radical operation from Feb 2010 to Aug 2013 were retrospectively analyzed.Of the 87 subjects, 31 subjects were given S-1 orally ( S-1 group) , 26 subjects given capecitabine ( capecitabine group ) orally and 30 not given any of them (control group).The data of CK20mRNA level of peripheral blood and positive rates of the three groups were tested and compared.Results The serum CK20mRNA positive rate in the S -1, capecitabine and control group were 51.6%( 16/31 ) , 46.2%( 12/26 ) and 83.3%(25/30), which suggested that the positive rate in control group was much higher than those in S-1 and capecitabine group ( P<0.05 ) , and there was no statistical difference found between S-1 group and capecita-bine group.The average serum CK20 mRNA expression level were (0.15 ±0.11), (0.14 ±0.09) and (0.89 ±0.46), respectively, which indicated that the data of control group was much higher than those in other two groups ( P<0.05 ) , and there was no statistical difference found between S -1 group and capecitabine group ( P >0.05 ). Conclusion Preoperative administration of S-1 and capecitabine in patients with colorectal cancer can significantly decrease the relative copy number and positive rate of CK20mRNA.

ObjectiveTo investigate the transcriptional changes of nitochondria fission and fusion gene loci and reactive oxygen species (ROS) level in cortical neurons of rats with chronic fluorosis,and to reveal their roles in mitochondria damage due to chronic fluorosis.MethodsSD rats were fed with different doses of fluoride through drinking water[＜ 0.5(control),10,50 mg/L,respectively] for 3 and 6 months.The level of ROS and mRNA contents of mitochondria fission gene loci Drp1/Fis1 and fusion gene locus Mfn1 in the cortical neurons of rat brains were detected with ROS fluorescent probe and real-time PCR,respectively.ResultsAs compared with control group [10.43 ± 5.98,(3.4 ± 0.6) × 103,(8.8 ± 1.4) × 10,(1.2 ± 0.2) × 102] at the experiment period of 3 months,the level of ROS and mRNA contents of mitochondria fusion gene locus Mfn1 and fission gene loci Drp1/Fis1 in the cortical neurons were obviously increased in the rats fed with 50 mg/L fluoride through drinking water[25.48 ± 6.09,(1.0 ± 0.2) × 1011,(3.0 ± 1.6) × 103,(8.9 ± 3.6) × 102,all P ＜ 0.05],whereas no significant changes were found in the rats fed with 10 mg/L fluoride[11.67 ± 3.49,(3.1 ± 0.3) × 104,(6.7 ± 2.7) × 10,(5.0 ± 0.9) × 10,all P ＞0.05].Furthermore,at 6 months of the experiment the increases in ROS level(63.02 ± 8.15,65.60 ± 7.40) and mRNA contents of mitochondria fission gene loci Drp1/Fis1 [(2.0 ± 0.8) × 106,(4.0 ± 0.6) × 105,(3.8 ± 1.3) × 103,(1.3 ± 0.2) × 103] and the decrease in mitochondrial fusion gene locus Mfn1[(3.0 ± 0.4) × 106、(4.0 ± 0.9) × 104]were observed in the cortical neurons of the rats fed with 10 mg/L and 50 mg/L fluoride as compared with the control group[25.26 ± 6.41,(3.0 ± 0.8) × 109,(5.1 ± 0.8) × 103,(2.8 ± 0.7) × 102,all P ＜ 0.05].Conclusions Excessive intake of fluorine leads to elevated ROS levels,and decreased transcription of mitochondrial fusion gene loci Mfn1,which is positively correlated with the time and dose-exposed to fluoride.The changes of mitochondrial fission and fusion gene loci in the cortical neurons may be related to high level of oxidative stress induced by chronic fluorosis.

Objective To investigate the deletion pattern of 4.8 kb mitochondrial DNA(mito-DNA) in liver,kidney,and brain of rats with chronic fluorosis and to explore the significance of mitochondria in the pathogenesis of chronic fluorosis.Methods Sixty SD rats were randomly divided into 3 groups according to body mass (20 in each group):control,low-fluoride and high-fluoride groups,and they were fed with different concentrations of fluoride in drinking water (0,10,50 mg/L,respectively) for 6 months.Mito-DNA in liver,kidney and brain was detected by real-time PCR.Results The amounts of 4.8 kb mito-DNA in liver(2.1 × 10-3,1.6 × 10-3),kidney (1.7 × 10-3,1.4 × 10-4) and brain cortex (1.5 × 10-5,1.3 × 10-5) in low-and high-fluoride groups were significantly reduced,as compared with that of control group (2.9 × 10-3,2.0 × 10-3,1.1 × 10-4,all P ＜ 0.05).The amount of 4.8 kb mito-DNA in kidney in high-fluoride group was lower than that in low-fluoride group (P ＜ 0.05).Conclusions Excessive fluoride intake can result in missing of 4.8 kb mito-DNA in liver,kidney and brain cortex.The abnormal of mito-DNA might be related to the dysfunction of mitochondrial respiratory chain.

ObjectiveTo explore the normal range of the volume of internal capsules in Chinese adults of the Han nationality and its relationship with age,body habitus,and craniocerebral volume.Methods One thousand healthy volunteers (age range =18 to 80 years) were divided into 5 groups according to their age;Group A ( 18 to 30 years old),group B (31 to 40 years),group C (41 to 50 years),group D (51 to 60 years),and group E (61 to 80 years).Each group consisted of 100 males and 100 females.MR imaging was performed in all of the volunteers using T1 weighted three-dimensional nagnetization prepared rapid acquisition gradient echo sequence. After three dimension data reconstruction,the volumes of bilateral internal capsules were manually measured. The volumes of bilateral internal capsules were compared by paired sample t test.The internal capsule volumes were compared between male and female by independent sample t test,and the differences among 5 age groups were compared by one-way ANOVA.The relationship between the volumes of internal capsule and age,body habitus or cerebral volume were analyzed using bivariate correlation.ResultsThe left and right internal capsule volumes were (2809 ± 393) and (2677 ± 343 ) mm3 respectively.The left internal capsule volumes were significantly larger than that of right (t =12.078,P ＜ 0.05 ).The left and right side of internal capsule volumes in male were (2863 ± 396) and (2744 ±358) mm3 respectively,and (2754 ±385) and (2609 ±314) mm3 in female.The left and right internal capsule volumes were larger in males than in female (t =1.982,2.851 ;P ＜ 0.05 ).The left internal capsule volume of the 5 age groups were ( 3273 ± 361 ),( 2943 ± 299 ),( 2777 ± 255 ),( 2607 ± 199 ),(2444 ±213) mm3,and the right were (2993 ± 361 ),(2814 ± 270),(2682 ± 239),(2543 ± 219),(2351 ±210) mm3.There were significant differences among 5 age groups between left and right internal capsule volume ( F =55.244,34.493 ; P ＜ 0.05 ).There was significant negative correlation between the volume of left and right internal capsule and age ( r =- 0.718,- 0.637 ; P ＜ 0.05 ).Conclusions1.5 T MR scanner can be used to accurately measure the internal capsule volumes.There is a significant negative correlation between age and internal capsule volumes.

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To evaluate the accuracy and value of measurement of left ventricular systolic function in dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM) patients with stereo three-dimensional echocardiography (S3 DE). Methods End-diastolic volume (EDV), end-systolic volume (ESV),stroke volume(SV) and ejection fraction(EF) of the left ventricle were measured with M-mode echocardiography(ME),two-dimensional echocardiography(2DE) and S3DE in DCM patientsC20 cases). HCM patients(20 cases),and normal controls(20 cases). The different results among the three groups or three methods were analyzed. Results (①In all the three groups,the results of EDV,ESV,and SV obtained with ME were significantly higher than those obtained with S3DE( P <0. 01). Only in normal group( P <0. 01) and HCM group ( P <0. 05) ,the results of EF obtained with ME and 2DE were significantly higher than that obtained with S3DE. ②By S3DE,compared with normal group,EDV,ESV were increased and EF was decreased obviously in DCM group (all P <0. 01); while in HCM group, only SV was significantly higher( P < 0. 01). ③EDV, ESV, and EF measured by S3DE were correlated and fit well with those measured by 2DE(r = 0.778,0.876, 0.932;R2 =0.605,0.767,0.869;all P <0.01). ④Within HCM group,excluding the impact of heart rate,cardiac output (CO) was highly correlated with SV( r = 0. 987,P < 0. 01). Conclusions S3DE can real-time display the stereo structure of the heart, and accurately and reliably assess the left ventricular systolic function, with a priority over traditional ME and 2DE methods. EDV,ESV, and EF are still effective indicators for the clinical assessment of left ventricular systolic function. SV obtained with S3DE will be expected to be the more sensitive and accurate value in assessing left ventricular systolic function in patients with early-stage cardiomyopathy.

OBJECTIVETo investigate the methods of preoperative diagnosis and differentiation of different pathological tissue in middle ear and mastoid.

METHODSThe temporal bone lamellar CT findings in 106 patients with chronic suppurative otitis media (including cholesteatoma) were retrospectively analyzed. The CT value of pathological tissue were measured for 183 times and were compared with the surgical findings and postoperative pathological findings to definitude the CT value range of different pathological tissue. Sixty patients taken from 106 patients at random were analyzed and made the diagnosis again by the same doctor team according to the CT value of the different pathological tissue and surrounding histoclasia resulted by pathological tissue. The diagnose accordance rate was compared with the routine diagnose report from radiology department. The predetective diagnosis was made in 10 patients with chronic suppurative otitis media according to clinical manifestation (pathological changes of tympanic membrane, nature of otorrhea, character of hearing), temporal bone lamellar CT finding (CT value of pathological tissue, surrounding histoclasia) to validate the value of this study for preoperative diagnosis and differentiation of different pathological tissue in middle ear and mastoid.

RESULTSThe CT value of cholesteatoma, granulation tissue, cholesteatoma combined with granulation tissue, effusion, calcified tissue, thickened and polypoid membrane was respectively (46.6 +/- 10.3) Hu, (26.6 +/-7.4) Hu, (42.1 +/- 11.4) Hu, (- 24.6 +/- 9.2) Hu, (223.6 +/- 63.7) Hu, (23.8 +/- 8.5) Hu. The diagnose accordance rate in 60 patients who were analyzed and made diagnosis again according to the CT value of the different pathological tissue and surrounding histoclasia resulted by pathological tissue raised from 68. 3% to 81.7% ( P < 0.05) . The predetective diagnose accordance rate reached at 90% according to clinical manifestation, temporal bone lamellar CT.

CONCLUSIONSIt was not reliable to diagnose and differentially diagnose different pathological tissue in middle ear and mastoid only by the CT value, however, the CT value could still be considered to be a very significant information. The accurate rates of diagnosis and differentiation of different pathological tissue in middle ear and mastoid obviously raised by synthetically analyzing various kinds of pathological tissues in middle ear and mastoid according to clinical manifestation, temporal bone lamellar CT finding.

Adolescent ; Adult ; Aged ; Child ; Chronic Disease ; Female ; Humans ; Male ; Mastoid ; diagnostic imaging ; Middle Aged ; Otitis Media, Suppurative ; diagnostic imaging ; Retrospective Studies ; Temporal Bone ; diagnostic imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo observe the mitochondrial fragmentation and the expression of mito-fusion 1 gene in the cortical neurons of rats with chronic fluorosis, and to reveal their roles in mitochondria damage to neurons due to chronic fluorosis.

METHODSSD rats were divided randomly into three groups of 20 each (a half females and a half males housed individually in stainless-steel cages), and fed with the different doses of fluoride containing in drinking water (untreated control containing 0 mg/L fluoride, and low-fluoride and high supplemented with 10 and 50 mg/L fluoride, respectively). After 3 or 6 months exposure, the mitochondrial morphology of the neurons in rat brains were observed by transmission electron microscopy (TEM), then the expression of mitochondrial fusion gene, Mfn1, were detected by immunohistochemistry and western-blotting, respectively.

RESULTSDental fluorosis was obvious in the rats exposed to excessive fluoride in their drinking water, that is, (16 rats out of 20) numbers of I° detal fluorosis in the low-fluoride group, and (11 rats out of 20) numbers of I° and (9 rats out of 20) numbers of II° detal fluorosis in the high-fluoride group were observed after 3 months exposure. Moreover, (14 rats out of 20) numbers of I° and (6 rats out of 20) numbers of II° detal fluorosis in the low-fluoride group and (6 rats out of 20) numbers of Io, (13 rats out of 20) numbers of II°, and (1 rats out of 20) numbers of III° detal fluorosis in the high-fluoride group were observed after 6 months exposure. And both of untreated controls without detal fluorosis were also observed. The urinary level of fluoride in the low-fluoride group (3.30 ± 1.18) mg/L and in the high-fluoride group (5.10 ± 0.35) were observed after 3 months exposure (F = 3.18, P < 0.05). Moreover, the urinary level of fluoride in the low-fluoride group (4.16 ± 1.39) mg/L and in the high-fluoride group (5.70 ± 1.70) mg/L were also observed after 6 months exposure (F = 3.17, P < 0.05). The normal mitochondrial morphology of neurons in rats without fluorosis was observed after 3 and 6 months, while the abnormal mitochondrial morphology of neurons with fluorosis was shown, presenting mitochondrial fragmentation with swollen cristae and even the fragmented, shortened or stacked punctuate membranes (section observation of three bullous mitochondrial-mitochondrial fission process) by TEM. As compared with controls (53.0 ± 4.54 and 1.21 ± 0.18) at the experiment period of 3 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 51.09 ± 6.25) and western-blotting (1.22 ± 0.26) were no significant difference for low fluoride group (t = 1.7, 1.1, P > 0.05); Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 59.71 ± 5.64) and western-blotting (1.66 ± 0.20) were significantly increasing for high fluoride group (t = 2.1, 2.1, P < 0.05). As compared with controls (36.43 ± 4.04 and 1.00 ± 0.13) at the experiment period of 6 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells 20.05 ± 4.55 and 17.10 ± 3.86) and western-blotting (0.64 ± 0.08 and 0.39 ± 0.06) were significantly decreasing for the two fluoride group (t = 2.1, 2.2; 2.2, 2.2 respectively, all P value were < 0.05).

CONCLUSIONSTaking excessive amount of fluoride might result in the mitochondrial fragmentation for the changed expression of Mfn1, and the neurons damage from the chronic fluorosis might be associated with the dysfunction of mitochondrial fusion.

Animals ; Drinking Water ; chemistry ; Female ; Fluoride Poisoning ; metabolism ; pathology ; Fluorosis, Dental ; metabolism ; Male ; Membrane Proteins ; metabolism ; Mitochondria ; pathology ; Mitochondrial Proteins ; metabolism ; Neurons ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley