Objective To observe the influence of coal burning fluorosis on learning and memory ability in rats and reveal its possible mechanisms. Methods Healthy 48 SD rats were divided into control, low-fluoride and high-fluoride group. All rats in fluoride exposed groups were fed with the eom polluted by drying processes with burning coal containing high level of fluoride obtained from the endemic fluorosis area to produce the animal model of fluorosis. The experiment period were 3,6 mouths, respectively. The ability of leaning and memory was measured by Morris test and cholinesterase activity detected by photometric method at 3 or 6 month after experiment, respectively. Results Fluoride contents signifieantlly influenced the escape latency, the numbers of crossing the platforms and the time of staying the platforms(the value of F was 29.29,6.47,6.50, respectively, P<0.01).In addition, the numbers of crossing the platforms and the time of staying the platforms were influenced by the exposed time(the value of F was 16.11,45.59, P<0.01). Furthermore, the fluoride contents and the exposed time had an interaction between the numbers of crossing the platforms and the time of staying the platforms (the value of F was 4.67,5.68, P<0.05 or<0.01). Three months after the experiment, the mean values of escape latency [(14.71± 4.85)s] of rats in highly fluoride exposed group were significantly prolonged as compared with controls [(9.28±4.22)s]; 6 month after the experiment, the mean values of escape latency[(12.42±8.03)s, (17.48± 8.05)s] of rats in both groups exposed to fluoride were significantly prolonged as compared to controls [(7.04± 3.29)s, P<0.05]. The decreased numbers of crossing the platforms[(1.62±0.87)number] and the declined time of staying the platforms[(16.70±5.02)s] were found in the rats exposed to high fluoride as compared to controls [(3.53±1.67 )number, (23.33±5.35)s, P<0.05]. The fluoride contents obviously influenced the activities of acetylcholinesterase and butylcolinesterase (the value of F was 12.83,13.27, P<0.01). On the other hand, the times of breeding also influnced the activities of butylcolinesterase (the value of F was 16.26, P<0.01). In 3 months of the experiment, the activities of butylcolinesterase [(0.55±0.12)kU/g] in low fluoride exposed group were significantly decreased in comparison with controls[(0.73±0.10)kU/g, P<0.05]. The activities of acetylcholinesterase[(0.62±0.42)kU/g] and butylcolioesterase[(0.58±0.10)kU/g] in high fluoride group were significantly decreased as compared to eontrois[(1.41±0.52), (0.73±0.10)kU/g, P<0.05]. The correlation analysis showed that there was a negative correlation between the cholinesterase and the escape latency(r=-0.68, P< 0.01), and a positive correlation between the cholinesterase and the time of staying the platforms(r=0.57, P< 0.01). Conclusions The ability of learning and memory in rats with coal buring fluorosis was decreased, which might be connected to the decreased activity of cholinesterase in a dose-effect correlation.

ObjectiveTo observe the inhibition effect of docetaxel-loaded lipid microbubbles (DLLM) combined with ultrasound targeted microbubbles destruction (UTMD) on microvessel in rabbit VX2 liver tumor models.MethodsSixty rabbits were randomly divided into 6 groups (n= 10),i.e.Doc group (used docetaxel only),DLLM group (used docetaxel-loaded lipid microbubbles),Doc+US group (used docetaxel combined with ultrasound positioning irradiation),PLM+US group (used microbubbles combined with ultrasound positioning irradiation),DLLM+US group (used docetaxel-loaded lipid microbubbles combined with ultrasound positioning irradiation) and control group.The expression of CD34 and VEGF and microvessel density (MVD) were compared among different groups.ResultsAfter treatment,the expression of CD34 in DLLM+US group was lower,the MVD of DLLM+US group was markedly lower than that of the other groups (P＜0.01),while the expression of VEGF in this group was the lowest among all 6 groups (P＜ 0.01).ConclusionDLLM combined with UTMD can inhibit the generation of microvessels in rabbit VX2 liver tumor,thus inhibit the growth of the tumor.

Objective To explore the relationship between fracture healing and level variation of nerve growth factor (NGF) in the serum of patients with femoral shaft fracture combined with traumatic brain injury. Methods Sixty-four patients with femoral shaft fracture were divided into simple fracture group (n=32) and composited injured group (fracture combined with traumatic brain injury, n=32). Treatments like open reduction and internal fixation, or fixation with unreamed tibial nail were performed according to the patient's condition. Two-site enzyme-linked immunosorbent assy (ELISA) was employed to detect the serum level of NGF 1, 7, 14 and 21 d after injury; and correlation analysis of clinical healing time of fracture and serum level of NGF was performed on the 1 d of injury in the composited injured group. The clinical healing time of fracture between the 2 groups were compared.Results The clinical healing time in the composited injured group (62.88±5.99) was shorter than that in the simple fracture group (82.47±3.07, t=-16.473, P=0.000); and there was a high negative correlation between the clinical healing time of fracture and serum level of NGF on the 1 d of injury in the composited injured group(r=-0.966, P=0.000). The serum level of NGF in the composited injured group was higher than that in the simple group at each time points (P＜0.05), and the serum level of NGF in the 2 groups peaked on the 14th d of injury; and decreased gradually. Conclusion Serum level of NGF increases in the early phase in patients with femoral shaft fracture combined with traumatic brain injury and remains at a high level for the first 2 weeks, which may play an important role in promoting fracture healing.

Objective To explore epithelial ovarian cancer(EOC)antigens that are potentially useful for cancer early detection and therapy.Methods A high quality cDNA library derived from ascites tumor cells of EOC patients(3 cases of serous EOC,1 case of mucinous EOC,and 1 case of endometrial carcinoma of ovary)was constructed,and the method of combining serological analysis of recombinant cDNA expression libraries(SEREX)and suppression subtractive hybridization(SSH)was used for screening cDNA library.All of the positive clones were sequenced and bioinformatics analysis with BLAST software in GenBank was performed.Serological mini-arrays of recombinant tumor antigens(SMARTA)was used to investigate the prevalence of autoantibodies to these antigens in both 96 ovarian cancer patients and 96 cancer-free controls.Results Fifty-five positive clones encoding different antigenic genes of EOC recognized by IgG and(or)IgM were obtained.It showed that these 55 clones derived from 45 distinct genes and these genes could be grouped into 6 classes as following according to homology with known expressed sequence tag(EST):(1)known ovarian carcinoma related genes:BARD1,et al;(2)homologous genes with other tumors:TM4SF1,et al;(3)homologous genes with special tissues:ILF3,FXR1,et al;(4) homologous genes with special function:TIZ,C1 D,et al;(5)embryo originating genes:PKHD1,et al; (6)novel genes:OV-189,et al.SMARTA results showed that the positive ratio of five EOC antigens TM4SF1(28% vs 9%),CID(21% vs 6%),BARD1(23% vs 5%),FXR1(23% vs 8%),OV-189 (31% vs 13%)which reacting with their IgG autoantibodies,three antigens TIZ(26% vs 8%),FXR1 (28% vs 11%),and OV-189(18% vs 7%)which reacting with their IgM autoantibodies in patients was higher than in controls(P

AIMIn order to get high-yield and high-quality cardiomyocytes from adult rat. We needed to clarify the optimal perfusion pressure in a constant flow condition and stopping digestion pressure.

METHODSThe rat was injected with heparin (1 000 IU/kg, i. p.) 20 minutes prior to the experimental protocol. The heart was excised and the aorta was cannulated rapidly. The cannulated heart was mounted on a Langendorff perfusion apparatus with constant flow and perfusion pressure was monitored. The initial perfusion pressure was maintained at 15 kPa or 10 kPa by regulating the flow rate. The heart was digested by 0.08 % collagenase I at 37 degrees C and the enzymatic digestion was terminated immediately when the perfusion pressure was lowered to 10 kPa, 7.5 kPa or 5 kPa.

RESULTSWhile the initial perfusion pressure was 15 kPa (n = 4), enzymatic digestion induced great extent increment in perfusion pressure (maximal pressure > 25 kPa). The left ventricle was enlarged significantly and the contractility of myocytes was decreased. While the initial perfusion pressure was kept at 10 kPa, the digestion only resulted in less extent increment in perfusion pressure (maximal pressure < 18.75 kPa). The left ventricle was enlarged slightly. In the same initial perfusion pressure, if the digestion was stopped at 10 kPa (n = 3), or 5 kPa (n = 4), myocytes appeared over much or insufficiency digestion, respectively. The viability of freshly isolated cardiomyocytes was less than 10%. The most of myocytes died after restoration of extracellular Ca2+ concentration. If the digestion was stopped at 7.5 kPa (n = 15). The viability of cardiomyocytes was 82.6% +/- 4.8% in fresh isolation, 30.4% +/- 4.5% after restoration of normal extracellular Ca2+ concentration, or 24.8% +/- 5.4% after 4 h standing. The most of rod-shaped cardiomyocytes were quiescent and had visible cross striations, sharp edges, and normal contractility.

CONCLUSIONWhen the initial perfusion pressure is kept at 10 kPa (low pressure) and the enzymatic digestion is stopped at 7.5 kPa, high yield and high quality cardiomyocytes can be obtained.

Animals ; Cell Separation ; methods ; Myocytes, Cardiac ; Perfusion ; Pressure ; Rats ; Rats, Sprague-Dawley

In order to culture cardiomyocytes or to observe the contractile function of adult mouse cardiomyocytes, it is necessary to isolate high-yield and high-quality cardiomyocytes at first. The mouse was injected with heparin (5,000 IU/kg, i.p.) 20 min prior to the experimental protocol, then was sacrificed by cervical dislocation. The heart was excised and the aorta was cannulated rapidly. The cannulated heart was mounted on a Langendorff perfusion apparatus with constant flow and perfusion pressure was monitored. The initial perfusion pressure was maintained at 40 mmHg by regulating the flow rate. The heart was digested by 0.05 % crude collagenase I at 37 degrees C and the enzymatic digestion was terminated immediately when the perfusion pressure was lowered to 28 mmHg. The heart was then cut off the cannula and the atria and aorta dissected away. The ventricular tissue was chopped and the single myocyte was dispersed gently by a wide tipped pipette. The viability of freshly isolated cardiomyocytes was more than 70 %. The cardiomyocytes were kept in Joklik's minimum essential medium containing 1 % BSA and 10 mmol/L BDM, then extracellular calcium was restored step-wise to a final concentration of 1.25 mmol/L. The viability of cardiomyocytes reduced to (40-50) % after 4 h standing. More than 90 % of rod-shaped cardiomyocytes were quiescent and had visible cross striations and sharp edges. The amplitude of unloaded shortening in cardiomyocytes was (9.72+/-0.43) % during 1.0 Hz stimulation, (11.28+/-0.43) % at 2.0 Hz and (11.40+/-0.45) % at 5.0 Hz. These results indicate that high yield and high quality cardiomyocytes can be obtained. In addition, the standards of identifying cardiomyocyte quality are concise and are suitable to culture the cardiomyocytes or to study the physiological function of cardiomyocytes.
Animals ; Cell Separation ; methods ; Cells, Cultured ; Female ; Male ; Mice ; Myocardial Contraction ; physiology ; Myocytes, Cardiac ; cytology ; physiology

OBJECTIVETo explore the differential proteomic expression in human liver cells L-02 after exposure to HQ.

METHODSSubcultured L-02 cells were treated by HQ for 24 h at a 1 x 10(-4) mol/L concentration and a blank group was set as the control. Immediately after the treatment, total cellular proteins were extracted and separated by 2-DE, and the images were analyzed by PDQuest software. The experiment was totally repeated 3 times with 3 repetitions for each group every time. The well repeated spots were identified by MALDI-TOF MS and then searched in NCBI human protein database with Mascot.

RESULTSAbout 1,000 spots per gel were found. Compared with the control group, 17, 18 and 24 spots were significantly altered in 3 separate experiments. The 4 well repeated spots were identified by MALDI-TOF MS as Rho GDP dissociation inhibitor GDI alpha, 6-phosphogluconolactonase, erbB3 binding protein EBP1 and lamin A/C, isoform 1 precursor. They were involved in cell skeleton, signal transduction and energy metabolization in functional classification.

CONCLUSIONHydroquinone can change the protein expression in liver cells, which provides clues for exploring the toxic mechanism.

Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Hepatocytes ; drug effects ; metabolism ; Humans ; Hydroquinones ; toxicity ; Mass Spectrometry ; Proteomics ; Reproducibility of Results

Objective To define the maximum-tolerated dnse(MTD)and observe the side effect of escalating cisplatin with 5-fluorouracil in concurrent chemoradiotherapy for esophageal carcinoma in Chinese,with toxicity studied.Methods Previously untreated fifteen Chinese patients suffering from esophageal carcinoma received conventional fractionafiun radiotherapy,with 5 daily fractions of 2.0 Gy per week.The total radiation dose was 60 Gy.Concurrent chemotherapy dose escalation was given by the relatively safe and kidney-sparing modified Fibonacci sequence.The starting dose was cisplatin 37.5 mg/m~2 D1 and 5-fluorouracil 500 mg/m~2 D1-5, respectively.This regimen was repeated 4 times every 28 days.Escalation dose was eisplatin 7.5mg/m~2 and 5- fluorouracil 100mg/m~2.Every cohort contained at least 3 patients.If no dose-limiting toxicity(DLT)was observed, the next dose level was opened for entry.These courses were repeated until DLT appeared.MTD was declared as one dose level below which DLT appeared.Results DLT was defined as grade 3 radiation-induced esophngitis at the level of cisplatin 60 mg/m~2,5-fluorouracil 700 mg/m~2.MTD was defined as eisplafin 52.5 mg,/m~,5- fluorouracil 700 mg/m~2.The major side effect were radiation-induced esophagitis,leucopenia,nausea,vomiting and anorexia.Conclusion Maximun tolerated dose of cisplatin with 5-fluorouracil in concurrent chemoradiotherapy in the Chinese people with esophageal carcinoma were eisplatin 52.5 mg/m~2 D1,5-fluorouracil 700 mg/m~2 D1-5,repeated 4 times every 28 days.

OBJECTIVETo explore the correlation of histologically proven prostatitis with the level of prostate specific antigen (PSA), prostate volume, PSA density (PSAD), international prostate symptom score (IPSS), maximum flow rate (Qmax) and post-void residual volume (PVR) in men with symptoms of benign prostate hyperplasia (BPH).

METHODSTotally 673 patients surgically treated for BPH were divided into Groups A and B in accordance with histological findings, the former including those with histological prostatitis, and the latter without it. Comparisons were made between the two groups in the PSA level, prostate volume, PSAD, IPSS, Qmax and PVR.

RESULTSThe PSA level, prostate volume, IPSS and PVR were significantly higher in Group A ([5.64 +/- 2.48] microg/L, [43.66 +/- 13.11] ml, 24.72 +/- 5.39 and [124.90 +/- 49.80] ml) than in B ([4.97 +/- 1.99] microg/L, [40.41 +/- 11.44] ml, 23.40 +/- 6.21 and [112.73 +/- 50.03] ml) (P<0.05), while Qmax markedly lower in the former ([6.94 +/- 3.23] ml/s) than in the latter ([7.75 +/- 3.52] ml/s) (P<0.05), but PSAD showed no statistically significant difference between the two groups (0.129 +/- 0.048 vs 0.123 +/- 0.034, P>0.05).

CONCLUSIONHistological prostatitis can significantly increase the PSA level, prostate volume, IPSS and PVR, and reduce the Qmax of the patient, but is not correlated with PSAD. It is an important factor influencing the clinical progression of BPH.

Aged ; Humans ; Male ; Organ Size ; Prostate ; metabolism ; pathology ; Prostate-Specific Antigen ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; urine ; Prostatitis ; metabolism ; pathology ; urine

Animals ; Fatty Liver, Alcoholic ; drug therapy ; Female ; Male ; Phytotherapy ; Rats ; Rats, Wistar ; Saponins ; therapeutic use ; Trigonella