Objective To establish a rat model of coal-burning-borne fluorosis,and to observe the expression changes of bone morphogenetic protein-2 (BMP-2) and bone morphogenetic protein-3 (BMP-3) in the serum of rat treated with different dose of fluoride and different treatment duration.Methods A total of 120 clean grade SD rats(body mass between 80 to 120 g) weaned for 4 weeks were randomly assigned into four groups,which were control,low-dose fluorid,medium-dose fluorid and high-dose fluorid groups,respectively,and 30 rats in each group (female 15,male 15).All of the rats were fed with coal drying corn from fluorosis area.Ten rats were killed by femoral artery bleeding 30 d,90 d and 180 d after exposed to fluoride,respectively.Serum BMP-2 and BMP-3 level was tested by enzyme-linked immunosorbent assay (ELISA).Results ①Results of BMP-2:after exposed to fluoride for 90 d and 180 d,the differences of serum BMP-2 level between groups were statistically significant(F=385.08,173.98,all P ＜ 0.01).In low-dose fluorid,medium-dose fluorid and high-dose fluorid groups,the expression of serum BMP-2 at 90 d[(18.80 ± 0.43),(22.22 ± 0.85),(25.14 ± 0.69)μg/L] and 180 d[(7.98 ± 0.68),(8.97 ± 0.78),(15.04 ± 0.89)μg/L] was higher than that of control group[(12.54 ± 1.29),(7.53 ± 0.97)μg/L,all P ＜ 0.05],and the level of BMP-2 increased with increasing dose of fluoride (all P ＜ 0.05).Within each group,the difference of serum BMP-2 was statistically significant(F =55.42,511.58,686.35,671.64,all P ＜ 0.01).The expression of BMP-2 in each group at 90 d [(12.54 ± 1.29),(18.80 ± 0.43),(22.22 ± 0.85),(25.14 ± 0.69)μg/L] was higher than that at 30 d[(11.75 ± 1.15),(11.42 ± 1.07),(11.38 ± 0.92),(11.15 ±1.03)μg/L,all P ＜ 0.05].The expression of BMP-2 in each group at 180 d[(7.53 ± 0.97),(7.98 ± 0.68),(8.97 ± 0.78),(15.04 ± 0.89) μg/L] was lower than that at 90 d.②Results of BMP-3:the difference between groups was not statistically significant at every experimental stage(F =0.7215,1.2951,0.0964,all P ＞ 0.05).Conclusions Longer excessive fluoride intake stimulates the expression of BMP-2 in rats,but with prolonged fluoride intake,the stimulation becomes weak.The effect of fluoride on BMP-3 is not as sensitive as that on BMP-2.

OBJECTIVETo study the molecular mechanism of curcumin in human esophageal carcinoma cell line (EC109).

METHODSEC109 cells were cultivated in vitro. When 80%-90% confluence was reached, they were treated with curcumin in different concentrations (15-120 µmol/L). The effects on cell proliferation were examined by CCK-8 colorimetry. The ultrastructure of EC109 cells were detected with transmission electron microscope(TEM). The cells apoptosis was observed with laser confocal microscope(LCM) by AnnexinV-FITC/PI double staining. The proteins level of PTEN, AKT, GSK3β and Caspase 3 were tested by flow cytometry(FCM) .

RESULTSCCK-8 test showed that curcumin could inhibit the proliferation of EC109 cells in a time- and concentration-dependent manner. TEM and LCM examinations indicated that curcumin could make EC109 cells apoptosis. The data of FCM showed that curcumin could increase the expression of PTEN, GSK3β and Caspase 3, decreased the expression of AKT.

CONCLUSIONThe effects of curcumin on inhibiting proliferation and promoting apoptosis of EC109 cells were related with increased expression of PTEN and inhibition of PI3K/AKT signaling pathway.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Curcumin ; pharmacology ; Esophageal Neoplasms ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction

Adolescent ; Adult ; Aged ; Creatine Kinase ; blood ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thymidine ; analogs & derivatives ; therapeutic use ; Young Adult

OBJECTIVETo explore the distribution, combination and evolution of various syndromic etiologies of erectile dysfunction (ED) based on the syndrome etiology theory.

METHODSUsing the ED Syndromic Etiology Scale, we collected the clinical data on the Chinese medicine diagnoses of 297 cases of ED, extracted the core syndromic etiologies by analysis of principal components and factors, and analyzed the patterns of distribution, combination, and evolution of ED syndromic etiologies according to the general information of the patients.

RESULTSThrough analysis of principal components and factors, 9 core syndromic etiologies were extracted, i. e. , liver constraint with qi stagnation, kidney yin deficiency, damp-heat, liver constraint transforming into liver-fire, blood stasis, kidney yang deficiency, heart-spleen paired deficiency, qi-yin paired deficiency, and phlegm-damp. Each of these syndrome etiologies exhibited its own specific distribution patterns. Of the total number of cases studied, 51.52% had 2 or 3 core syndromic etiologies and 36.03% had only one.

CONCLUSIONIn the early stage of ED, its syndromic etiologies are usually liver constraint with qi stagnation, kidney yin deficiency, damp-heat, liver constraint transforming into liver-fire, and blood stasis. With the natural progres- sion of the disease, its syndromic etiologies gradually evolve into kidney yang deficiency, heart-spleen paired deficiency, qi-yin paired deficiency, phlegm-damp, and blood stasis, and finally into yin-yang deficiency of the heart, spleen and kidneys, combined with phlegm-damp and blood stasis.

Adult ; Erectile Dysfunction ; diagnosis ; drug therapy ; etiology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged

OBJECTIVETo investigate the inhibition effect of curcumin on the proliferation of the human esophageal carcinoma cell line Ec109 and its impact on PEN/PI3K/Akt signaling pathway.

METHODSEsophageal carcinoma Ec109 cells were cultured in vitro conventionally and were treated with curcumin at different concentrations. The cell proliferation level was examined by MIT colorimetry, the ultrastructure of curcumin-treated Ec109 cells were detected with transmission electron microscope (TEM) and cell apoptosis was observed by FCM with AnnexinV-FITC/PI double staining. The protein levels of PTEN, Akt, GSK3P and Caspase 3 of curcumin-treated Ec109 cells were detected by Western blot.

RESULTSMTT test showed that curcumin could inhibit the proliferation of Ec109 cells in a time and concentration-dependent manner. TEM examination indicated that curcumin could induce Ec109 cell apoptosis. FCM detection showed that Ec109 cell apoptotic rate increased significantly with the increase of drug concentration. On the other hand, curcumin could promote the expression of PTEN, GSK3beta and Caspase 3 yet reduce the expression of Akt.

CONCLUSIONCurcumin could obviously up-regulate the expression of PTEN, GSK3beta and Caspase 3, surpress PI3K/Akt signaling pathway and hence inhibit the proliferation of Ec109 cells.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Curcumin ; pharmacology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Oncogene Protein v-akt ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; drug effects

The present study was designed to observe the influence of cerebral ischemia/reperfusion injury on learning and memory in hyperlipidemic rats and estimate the changes of activity of autonomic nervous system. Twenty-three male Wistar rats were randomly divided into three groups, named control group (C group, n=10), hyperlipidemia group (H group, n=6) and hyperlipidemia-ischemia group (HI group, n=7), respectively. The rats in H and HI group were fed a high-fat diet for 2 weeks and the rats in all groups were examined through Morris water maze (MWM) task. The rats in HI group underwent ischemia/reperfusion by 2-vessel occlusion (2-VO) method, and had electrocardiogram (ECG) recording simultaneously. The MWM task and ECG recording were taken again after 7 d of recuperation. The following results were obtained: (1) In the second place navigation performance and probe trial performance, the frequency of memory in quadrant of hidden-platform and memory score decreased significantly in HI group compared to that in C and H groups. (2) The heart rate in HI group decreased slowly after ischemia; the power at high frequency band (HF) reduced gradually, meanwhile the power at middle frequency band (MF) and the ratio of power at MF and HF decreased clearly compared to baseline value. (3) After 7 d of ischemia/reperfusion, the heart rate in HI group was significantly higher than that in H group (P<0.05). While there was no statistical change in the power at MF, the power at HF decreased and the ratio of MF/HF increased significantly (P<0.05). The data demonstrated that ischemia/reperfusion decreased the activity of autonomic nervous system, and the reduction of sympathetic nerve activity was much more than that of vagus nerve activity. The results suggest that the hippocampus neuron injury caused by ischemia induces cognitive disorder and imbalance of vago-sympathetic nerve activity accompanied by vagus nerve suppression.
Animals ; Autonomic Nervous System ; physiopathology ; Brain Ischemia ; etiology ; physiopathology ; Heart Rate ; physiology ; Hippocampus ; physiopathology ; Hyperlipidemias ; complications ; Learning Disorders ; etiology ; physiopathology ; Male ; Memory Disorders ; etiology ; physiopathology ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; physiopathology

OBJECTIVETo observe the hair follicle regeneration after subcutaneous implantation of hair follicle cell mixture in nude mice.

METHODSThe hair papilla cells, dermal sheath cells, outer root sheath and fibroblasts of human scalp were mixed with the hair follicle epithelial cells and implanted subcutaneously in nude mice to observe the regeneration of the hair follicle.

RESULTS AND CONCLUSIONFormation of intact hair follicle-like structures was observed in the skin sections of the recipient nude mice, suggesting the feasibility of this approach for hair follicle regeneration in vivo.

Animals ; Cell Transplantation ; Hair Follicle ; cytology ; metabolism ; physiology ; transplantation ; Humans ; Injections, Subcutaneous ; Mice ; Mice, Nude ; Regeneration ; Skin Pigmentation ; Time Factors

OBJECTIVETo gain stable genetic modification of neural stem cells (NSC) that constitutively secrete brain-derived neurotropic factor (BDNF) and to explore the biological role on the survival and neurite outgrowth of cultured dorsal root ganglion (DRG) neurons.

METHODSBDNF gene fragment from human brain cDNA libraries was obtained by using PCR. With molecular cloning technique, the recombinant stem cell viral vector with report gene was constructed, which is that MSCV-BDNF-IRES(2)-EGFP vector could encode Polycistronic mRNA. Viral particle was packaged by PT67 cell line and infected neural stem cells (mouse clone: C17.2). After selection with cloning cylinder, the expression of BDNF was assessed by immunohistochemistry enzyme linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). The effects of stable gene-modified neural stem cells on embryonic mouse DRG neurons were evaluated in a dual-chambered cocultivation system at 3th, 10th day.

RESULTSRT-PCR analysis demonstrated expression of mRNA for human-BDNF. ELISA confirmed the presence of secreted BDNF 24 h after transfection and showed that the level of BDNF production by NSC-BDNF transfected C17.2 was at a rate of (14.6 +/- 0.8) ng x d(-1) x 10(-6) cells even after 3 months. With immunohistochemical analysis, compared with the control, the longer neurite outgrowth of cultured DRG cells and the more survival neurons were observed in NSC-BDNF transfected cells group.

CONCLUSIONSBDNF gene could be stably expressed in C17.2 cell line by MSCV, and the BDNF gene-modified NSC markedly enhances the survival and neurite outgrowth of cultured DRG neurons.

Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Cell Culture Techniques ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Ganglia, Spinal ; cytology ; Humans ; Mice ; Neurons ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; metabolism ; Transfection

OBJECTIVETo study the X-ray and CT findings of traumatic bronchial rupture for early radiographic diagnosis and treatment.

METHODSThe chest plain X-ray films and CT images of 21 patients with traumatic bronchial rupture confirmed by operations or bronchoscopy were retrospectively analyzed.

RESULTSThe main radiographic findings of traumatic bronchial rupture included interrupted tracheobronchial air column, atelectasis, lung ptosis, pneumomediastinum and subcutaneous emphysema, pneumothorax or hydropneumothorax. CT scanning also revealed tracheobronchial wall defect, bronchostenosis, and bronchial occlusion, displacement and angulation.

CONCLUSIONChest plain X-ray film combined with CT scanning has important values for early diagnosis of traumatic bronchial rupture.

Adolescent ; Adult ; Aged ; Bronchi ; injuries ; surgery ; Bronchoscopy ; Early Diagnosis ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Rupture ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo explore the relationship between α-actinin content and cardiac function in rats during myocardial ischemia-reperfusion.

METHODSThirty-two rats were randomized equally into sham-operated group, 30 min ischemia group, 1 h ischemia group, and 1 h ischemia with 2 h reperfusion group. Acute myocardial ischemia was induced in the 3 ischemia groups by ligation of the left anterior descending coronary artery, and the cardiac functions were evaluated. The myocardial contents of α-actinin was measured by immunohistochemistry, and phospholipase C (PLC) and phosphatidylinositol-3-kinase (PI3K) contents were determined by ELISA after the operations.

RESULTSThe left ventricular systolic pressure (LVSP), +dp/dt max, and -dp/dt max tended to decrease during myocardial ischemia, and increased after reperfusion, and the left ventricular end-diastolic pressure (LVEDP) showed reverse changes. The levels of α-actinin decreased with prolonged ischemia, showing a significant difference in 1 h ischemia group from those in the other 3 groups. PI3K and PLC contents were significantly increased with prolonged myocardial ischemia. Stimulation by LY-294002 and U-73122 caused enhanced contraction of single cardiomyocytes, and also increased the fluorescence intensity of α-actinin in the cardiomyocytes compared with that in 1 h ischemia group.

CONCLUSIONSThe cardiac dysfunction during acute ischemia-reperfusion in rats may be related with the changes of myocardial α-actinin content, which are probably a result of increased PI3K and PLC contents in the ischemic myocardium.

Actinin ; metabolism ; Animals ; Myocardial Ischemia ; metabolism ; physiopathology ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Myocardium ; metabolism ; Phosphatidylinositol 3-Kinase ; metabolism ; Rats ; Rats, Wistar ; Type C Phospholipases ; metabolism