Objective: To observe the influence of spent culture supernatant (SCS) of Lactobacillus acidophilus strain LA14 on the contraction of isolated intestinal smooth muscle and to discuss the related mechanism. Methods: The ileum samples of rabbits were prepared and the contraction frequency and amplitude of intestinal smooth muscle were observed as the normal control. Then the SCS, bacterium suspension, and SCS with bacterium suspension were added by an accumulative dose to the culture media (0.3 ml per times, at an interval of 6 min), respectively. Four minutes after each administration, the contractive curves were recorded for 2 min. The influences of various groups of Lactobacillus acidophilus on the contraction of isolated intestinal smooth muscle were observed. The effect of SCS on M cholinoceptor was observed by adding in order pilocarpine, atropine or SCS, and pilocarpine. Results: After continuous administration of SCS or SCS with bacterium suspension (0.6-1.5 ml), the contraction frequency of the intestinal smooth muscle was significantly lowered compared with before administration (P<0.05 or P<0.01), and there were no significant differences between before administration and other volume groups (P>0.05). Within the range of 0.3-1.5 ml, the SCS, bacterium suspension, and SCS with bacterium suspension resulted in no significant difference in reducing the contraction amplitude, except for SCS with bacterium suspension at 1.5 ml(P<0.05). SCS or atropine significantly inhibited pilocarpine-induced increase of contraction amplitude(P<0.05 or P<0.01). SCS also reduced the contraction frequency of the intestinal smooth muscle(P<0.01). Conclusion: SCS of Lactobacillus acidophilus may inhibit the peristalsis of the intestinal smooth muscle of rabbits by blocking M cholinoceptor.

Child, Preschool ; Humans ; Hypercalcemia ; etiology ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

Objective To perform comparative proteomic analysis of human ovarian cancer cell lines for detecting platinum-resistance associated proteins.Methods The total proteins of two sensitive (SKOV3 and A2780)and four resistant(SKOV3/CDDP,SKOV3/CBP,A2780/CDDP and A2780/CBP) human ovarian cancer cell lines were separated by two-dimensional gel electrophoresis(2-DE).The differentially expressed proteins were analyzed using image analysis software,stained with Coomassie Brilliant Blue,then identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and database searching.The mRNA and protein levels of the differentially expressed protein which was most significant in all of the four resistant cell lines were validated by RT-PCR and western blotting,respectively.Results Five proteins were found to be significant in four cell lines. Annexin A3 and destrin were up-regulated and nicotinamide-adenine dinucleotide phosphate(NADP)- dependent isocitrate dehydrogenase 1 was down-regulated in all the four resistant samples.Glutathione transferase omega 1 had an increased expression in the other three resistant cell lines except for SKOV3/CBP in which its expression was not changed.However,cofilin 1 represented a different trend.In the two resistant sublines of SKOV3,eofilin 1 had a down-regulation,but it had an up-regulation in the cell lines induced from SKOV3.The expression of annexin A3 was up-regulated by 3-20 fold and the results of RT- PCR and western blotting showed complete consistency with that by 2-DE.Conclusions Proteomic techniques are useful to the identification of the resistance-associated proteins in ovarian cancer platinum- resistant cell lines and five candidates have been found.The five differential proteins might become hopeful candidate biomarkers for resistance.

Background Scarring of surgical area,the most important factor,leads to the failure of glaucoma filtering surgeries.Therefore,more and more attentions are paid to the causes and process of scar formation.Objective This study was to compare the differences of proliferation and migrating abilities of fibroblasts between filtering bleb scar tissue and normal Tenon capsular tissue,and to investigate the inhibitory effects of miRNA-200a (miR-200a) on biological behavior of conjunctival fibroblasts.Methods Normal Tenon capsular tissue and filtering bleb scar tissue were collected during the strabismus surgery and glaucoma filtering surgery,respectively for the primarily culture of fibroblasts.The proliferation (absorbency,A) of the cells was assayed by cell counting kit-8 (CCK8) method;the relative migrating distance of the cells was measured by cell scratch test;and the relative expressions of transforming growth factor-β1 (TGF-β1)mRNA and miR-200a mRNA in the cells were detected by realtime fluorescence quantitative PCR.TGF-β1 mimic of 0,1,2 and 5 ng/ml was added in the medium of human normal Tenon capsular-derived fibroblasts (HTFs),and 0.00,0.25,0.50,1.00 μg/ml TGF-β1 inhibitor was added in the medium of human scarring-derived fibroblasts (HSFs) for 24 hours,respectively,and CCK8 was used to evaluate the proliferation of the cells.The relative migrating distance as well as the relative expressions of miR-200a mRNA were analyzed in the 2 ng/ml TGF-β1 mimic-or 1.00 μg/ml TGF-β1 inhibitor-treated cells.Results The primary conjunctival presented the spindle and star-like in shape with large body and oval nuclei.The cells showed the positive response for keratin and vimentin antibodies.The A values were 1.476±0.110 in the HSFs and 0.958±0.074 in the HTFs,with a significant difference between them (t =24.900,P=0.016).The relative expressions of TGF-β1 mRNA were significantly higher in the HSFs than those in the HTFs,and the relative expressions of miR-200a were evidently lower in the HSFs than those in the HTFs,showing significant differences between them (t =6.358,P =0.024;t=7.394,P =0.018).Compared with the 2 ng/ml TGF-β1 mimic-treated HTFs,the relative migrating distance increased,while the expression level of miR-200a mRNA was significantly reduced in the 2 ng/ml TGF-β1 mimictreated HSFs (all at P＜0.05);Compared with the 1.00 μg/ml TGF-β1 inhibitor-treated HTFs,the relative migrating distance decreased,but the expression level of miR-200a mRNA was significantly elevated in the 1.00 μg/ml TGF-β1 inhibitor-treated HSFs (all at P＜0.05).Conclusions The proliferation and migrating abilities are stronger in the HSFs than those in the HTFs,which probably is regulated by the expression of miR-200a in the cells.The miR-200a plays a negative feedback for the effect of TGF-β1 promoting proliferation and migration of fibroblasts.

Recent work has shown that nitric oxide (NO) induction by nitric oxide synthase(NOS) is the physiological mediator of bone cell function and demonstrated that it may be possible to exert differential effects on osteoblast (OB) and osteoclast (OC) activity in vivo. The proinflammatory cytokines, such as tumor necrosis factor-? (TNF-?), interleukin-1? (IL-1?), interferon-? (IFN-?), will stimulate bone resorption by NOS-induced low levels of NO. These findings confirm NO as a potentially important osteoblast-osteoclast coupling factor, indicating that cytokine-induced NO was largely responsible for the mechanisms of osteoporosis. Pharmacological modulation of NO may therefore represent a new approach in the treatment of bone diseases characterized by increased bone resorption, such as osteoporosis (OP).

Acupressure ; Acupuncture, Ear ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Middle Aged ; Phytotherapy ; Postmenopause ; Premenopause ; Syndrome

In order to improve the expression level of segment of GABAA receptor a1 subunit in E. coli, growth conditions of the recombatant, which influence the final yield of protein expression, including growth medium, inoculation ratio, temperature, pH, rotation speed, inducing time and concentration of IPTG and so on, were studied in shaking flasks. The results indicated that, with 3% inoculation ratio, cultured 3.5 hours at 37℃, and then induced 5 hours by IPTG at 32℃, the yield of GABAA receptor protein was 95mg/L and the biomass was 3.25 g/ L. In contrast, using a 16 L stirred fermentor instead of shaking flasks, the highest level of the protein expression, 136mg/L with 4.95g/L of biomass, was achieved after fermenting 5.5 hours.

Adult ; Dimethylformamide ; poisoning ; Female ; Humans ; Occupational Exposure ; Young Adult

Objective To observe the changes of the intercellular spaces of squamous epithelium of lower esophagus in gastroesophageal reflux disease(GERD).Methods Eleven outpatients with GERD symptoms more than 3 months [6 with nonerosive reflex disease(NERD)and 5 with erosive esophagitis(EE)]and 5 healthy volunteers were recruited.All of them underwent endoscopy and 24-hr ambulatory pH monitoring.Biopsies were taken in lower esophagus(2 cm above Z-line)for electron microscope examination.Results Intercellular spaces of esophageal epithelial cell in volun teers,NERD patients and EE patients were (0.374?0.073)?m,(1.308?0.079)?m and (1.332?0.144)?m respectively,with significant differences between the control group and the NERD or EE group.There was no difference between NERD group and EE group.Conclusions Dilated intercellular spaces were seen in both NERD and EE cases,which was significantly different from the control cases.

AIM:To discuss the related factors that affected the stability of posterior corneal surface after laser in situ keratomileusis ( LASIK) .
METHODS:About 64 patients (64 eyes) were enrolled. The correlation among the changes in posterior corneal surface 6 month after LASIK, surgery method, corneal flap thickness ( FT ) , ablation thickness ( AT ) , postoperative residual corneal stroma thickness ( RCST ) , preoperative thinnest corneal thickness ( CT ) , flap thickness/preoperative thinnest corneal thickness ( FT/CT ) , ablation thickness/preoperative thinnest corneal thickness ( AT/CT) , postoperative residual corneal stroma thickness/preoperative thinnest corneal thickness ( RCST/CT) , anterior and posterior preoperative corneal height, the difference of the forward shift in posterior corneal surface ( diff value ) of preoperative and preoperative intraocular pressure were analyzed.
RESULTS: The changes of diff value between preoperative and postoperative were related with diopter (r=0.419, P=0.014), AT (r=0.394, P=0.023), AT/CT (r=0.501, P=0.004), Diff value of preoperative (r=0.501, P=0. 004), RCST (r=-0. 385, P=0. 033) and RCST/CT (r=-0. 401, P=0. 025). The changes of height value from posterior corneal surface between preoperative and postoperative were related with diopter (r=0. 520, P=0. 002), AT (r=0.504, P=0. 003), AT/CT (r=0. 442, P=0. 013), Diff value of preoperative (r=0. 624, P=0. 000) and RCST/CT (r=-0. 394, P=0. 028).
CONCLUSION: AT, RCST, AT/CT, RCST/CT and diff value of preoperative should be the key index that predicted the stability of posterior corneal surface after LASIK,the further research will give the range of safety value.