BACKGROUND:In the early development of zebrafish embryos, cels divide and proliferate rapidly, but low concentration of deguelin can delay the development of zebrafish embryos.
OBJECTIVE:To observe the effect of different concentrations of deguelin on cyclin D1 gene expression in zebrafish embryos.
METHODS:Though normal fertilization, zebrafish embryos that incubated to the 2-cel stage (about 0.75 hour after fertilization) and shield stage (6 hours after fertilization) were colected and put into 12-wel plates treated with 100, 200, 400 nmol/L deguelin at 28.5℃in an incubator til the shield period and 24 hours after fertilization, respectively. Simultaneously embroys treated with 1% dimethyl sulfoxide solution were as a control group, cultured in the same conditions. Cyclin D1 RNA probes were prepared for the whole mountin situhybridization, observing staining by an upright fluorescent microscope camera to detect the effect of deguelin on cyclin D1 expression in zebrafish embryos.
RESULTS AND CONCLUSION:Deguelin showed significant negative regulation on the expression of cyclin D1 gene in zebrafish embryos. Cyclin D1 expressed in outsourcing cels in embryos of shield stage, and a significant reduction in the expression of cyclin D1 came up with the increasing concentrations of deguelin. In the 400 nmol/L deguelin treatment group, there was nearly no expression of cyclin D1. Cyclin D1 expressed in the brain, central nervous system, immature eye, somites, trunk, and tail of embryos at 24 hours after fertilization, and reduced significantly in the 100 nmol/L deguelin treatment group, especialy in the proliferative area. In the 200 and 400 nmol/L treatment groups, the embryonic development slowed down signficantly, and cyclin D1 gene mainly expressed in the dorsal ectoderm cels.

· AIM: To evaluate the safety of eye anterior tissue when plasmin (Pm) and hyaluronidase (HS) are injected into pigs'vitreouses to induce posterior vitreous detachment(PVD).· METHODS: 15 pigs without ocular diseases were randomly assigned to groups A,B,C (5 in each group). For each pig, one eye was experimental, the other eye was control. The experimental eye received intravitreal injection with enzyme:group A: 50U(0.1mL) HS; group B: 0.5U(0.1mL) Pm; group C: 0.5U(0.05mL) Pm combined with 50U(0.05mL) HS; while the control eye received intravitreal injection with equivalent dose of balanced salt solution (BSS). Postoperative reactions in the eyes were carefully observed by clinical examinations such as slit-lamp microscopy and direct and indirect ophthalmoscopy and measurement of intraocular pressure (IOP). Eyeballs were extirpated 7 days after the operation and histological examination was carried out. The corneas and irises were observed under light microscope (LM). The epithelia of ciliary bodies and lens were studied with transmission electron microscope (TEM).· RESULTS: In all experimental eyes and control eyes of group A, B and C, histological examination showed: under LM ,the corneas and irises of experimental eyes had no histological abnormality in structure; Under TEM, epithelial cells of ciliary bodies and the lens were tightly arrayed with clear cellular boundaries, clear intracellular structure, intact cell membranes, and intact nuclei; There was no significant difference in the clinical examinations and in the change of IOP between preoperation and postoperation, between the experimental group and the control group.· CONCLUSION: With 0.5U Pm and HS 50U injected into pigs' vitreouses to induce PVD, alone or combined with two enzymes, evaluation revealed no evidence of toxicity on the eye anterior tissue. The dosage was proved to be safe for eye anterior tissue.

· AIM: To investigate the efficacy and safety of intravitreal injection of plasmin, hyaluronidase, or the combination of the two in inducing posterior vitreous detachment (PVD).· METHODS: 15 mini-type pigs were assigned to three groups (Group A, B and C), 5 in each group. One eye of each pig was intravitreally injected with the studying agent,and the fellow eye was used as control. Group A received a vitreous injection of hyaluronidase 50U (0.1mL); group B received plasmin 0.5U (0.1mL); group C received plasmin 0.5U (0.05mL) combined with hyaluronidase 50U (0.05mL). The fellow eyes in each group were injected with 0.1mL balanced salt solution (BSS). All the pigs were examined with slit-lamp biomicroscope, direct and indirect ophthalmoscope, B-scan and electroretinograph (ERG). After 7 days, the animals were killed and the eyes were enucleated and examined with light microscope, transmission electron microscope and scanning electron microscope.· RESULTS: B-scan examination showed that one eye of Group A and two eyes of Group B had partial PVD at 1st day after injection and one eye of Group C at 1 hour after injection. On the 7th day, B-scan, light microscopy and scanning electron microscopy demonstrated that all the eyes of Group A and Group B had partial PVD, while none of the control eyes had PVD. Rank sum test for scanning electron microscopy results of all the groups showed P ＜0.005.Furthermore, the comparisons between every two groups were made. The results of analyses were as follows: P＞0.05 between the drug injected eyes of Group A and Group B, P＜0.05 between Group B and Group C, Group A and Group C.The b-wave and a-wave amplitudes of ERG showed no significant difference either between preinjection and postinjection in all groups or between the drug injected eyes and the control eyes in each group. Light microscopy and transmission electron microscopy revealed no damage to the retinal structure.· CONCLUSION: Intravitreal injection of hyaluronidase 50U or plasmin 0.5U or their combination can produce PVD effectively and quickly without retinal functional or structural toxicity. The combination of the two proteases was proved to be synergetic.

Objective To investigate the reversal effect and molecular mechanisms of NF-κB inhibitor parthenolide (PTL) on insulin resistance ( IR). Methods HepG2 cells were treated with insulin at different concentrations and time points, the glucose consumption of HepG2 cells was measured via glucose oxidase method to determine the best concentrations and time for establishing insulin-induced insulin resistance on HepG2 cells. After modeling, different concentrations of PTL were added in cells for 24 h for determining cell activity and glucose-consumption. Q-PCR was used to detect the expression of NF-κB and IκBα mRNA in cells, and western blot was used to detect the expression of protein IκBα. Results The best reaction time and concentration for insulin inducing resistance of HepG2 cells were 24 h and at 10 μg·mL-1 . The optimum acting dose of PTL was 20 μmol·L-1 . NF-κB activity was significantly reduced (P<0. 05), IκBα degradation was significantly inhibited (P<0. 05) compared to HepG2 cells with insulin resistance upon intervention on insulin resistance HepG2 cells by PTL. Conclusion PTL can inhibit IκBα degradation and disassociation of it from NF-κB, which in turn improves insulin resistance, providing theo-retical basis for preventing and treating diabetes with PTL.

Administration, Inhalation ; Adrenal Cortex Hormones ; pharmacology ; Asthma ; drug therapy ; metabolism ; Child ; Child, Preschool ; Drug Interactions ; Drug-Related Side Effects and Adverse Reactions ; Female ; Humans ; Male ; Theophylline ; pharmacokinetics ; pharmacology ; Treatment Outcome

Objective To investigate the roles of theaflavins(TFs) in airway mucus hypersecretion induced by human neutrophil elastase(HNE). Methods Human lung adenocarcinoma cell A549 was stimulated by HNE for mucus hypersecretion,and TF monomers(TF1,TF2 and TF3) were used for intervention.The effects of TF monomers on viability of A549 cells were examined by MTT method.After the effective doses of TFs were determined,A549 cells were divided into 4 groups for experiment.In control group,A549 cells were cultured with serum-free medium.In HNE treatment group,A549 cells were treated with HNE(50 nmol/L) for 24 h.In TF monomer intervention groups,A549 cells were pre-treated with TF1,TF2 or TF3(50,100 or 200 ?g/mL) for 24 h,and were then treated with HNE for another 24 h.In AG1478 intervention group,A549 cells were pre-treated with AG1478(5 ?mol/L),an epidermal growth factor receptor blocker for 30 min,and were then treated with HNE for another 24 h.The changes in mucin(MUC) after treatment by different doses of TF1,TF2 and TF3,and by TF3(200 ?g/mL) for different time(12 h,24 h and 36 h) were detected.The changes in MUC5AC mRNA expression and MUC5AC protein expression were detected by RT-PCR and ELISA,respectively. Results The MUC5AC mRNA expression and protein expression in HNE treatment group were significantly higher than those in control group(P

Objective: To observe the efifcacy and safety of bivalirudin on primary percutaneous coronary intervention (PCI) in patients with acute ST-elevation myocardial infarction (STEMI).
Methods: A total of 159 patients with acute STEMI treated by emergent PCI in our hospital from 2011-09 to 2014-01 were retrospectively studied. The patients were divided into 2 groups according to procedural bivalirudin application as Bivalirudin group and Heparin group, and the application of GPI (glycoprotein IIb/IIIa inhibitor) was decided by the operator. The baseline condition, coronary artery imaging condition, peri-operative and 30-day post-operative bleeding, the occurrence rate of MACE were compared between 2 groups.
Results: There were 153 patients completed the follow-up study including 72 in Bivalirudin group and 81 in Heparin group. The peri-operative bleeding rates in Bivalirudin group and Heparin group were 6.5% vs 11.0%, the in-stent thrombosis rates were 0% vs 1.2%, 30-day post-operative bleeding rates were 9.7% vs 13.5% and the occurrence of MACE were 1.4% vs 7.4% allP>0.05.
Conclusion: THE application of bivalirudin in emergent PCI is safe and effective in patients with acute STEMI, it has certain trend to reduce bleeding in relevant patients.

· AIM: To explore the effects of C3F8 tamponade on hypotony on or after primary operation and the prognosis of severe ocular trauma.· METHODS: Twenty-six cases (26 eyes) of severe ocular trauma were treated with pure C3F8 tamponade on or after primary operation. IOP was observed, and the curative effect of C3F8 tamponade was observed on secondary operation with prognosis evaluated.· RESULTS: Hypotony improved in 23 eyes postoperatively,in which 18 eyes with edematous and cloudy cornea, 15 eyes had clear cornea after gas tamponade. Retina was reattached under the gas action in 21 eyes during the secondary operation. Visual acuity improved in 22 eyes, remained unchanged in 3 eyes and decreased in 1 eye during the follow-up of 3-12months.· CONCLUSION: Application of pure C3F8 tamponade on or after primary operation can effectively improve hypotony after severe ocular trauma and benefit a better prognosis.

OBJECTIVE: To study ApoB gene genetic polymorphism of Han nationality and Mongolian nationality in midwest area of Inner Mongolia. METHODS: Some unrelated individuals of Han nationality and Mongolian nationality in midwest area of Inner Mongolia were selected. Polymerase chain reaction-restriction fragment length polymorphism technology was used to check the presence of Xba I (X+) and EcoR I (E-) sites of rare alleles. The genotype frequency, allelic frequency and population genetics parameters were calculated. RESULTS: The frequencies of Xba I (X+) and EcoR I (E-) rare alleles were 2% and 4.6% in Han population. There was no Xba I (X+) or EcoR I (E-) rare alleles found in Mongolian nationality. CONCLUSION The allelic frequencies of ApoB gene Xba I and EcoR I sites are very different in different races. These sites may be used in identification of ethnicity.
Alleles ; Apolipoprotein B-100/genetics* ; Asian People/genetics* ; China ; Ethnicity ; Gene Frequency ; Genotype ; Humans ; Mongolia ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

Objective To study the immunoregulatory effects of bone marrow mesenchymal stem cells (MSC) on allogeneic peripheral B lymphocytes in vitro. Methods MSCs were isolated and cultured from bone marrow by gradient centrifugation. Mononuclear cells were isolated routinely from peripheral blood, then monocytes were eliminated by L-leucine methy ester method. Remained T lymphocytes were eliminated by AET-SRBC rosette method. The action of MSCs and its supernatant on B lymphocytes proliferation in the presence of anti-human IgM goat antibodies (anti-IgM) was investigated by MTT. The IgG, IgM in the supernatant were detected by ELISA. The percent of apoptosis B lymphocytes, co-cultured with MSCs for 24 or 48h, was assayed by FACS. Results MSCs and its supernatant inhibited B lymphocytes proliferation and Ig secretion. The inhibitory effect depended on the amount of MSCs and condition of its supernatant. The date of FACS indicated that the apoptosis ratio of B lymphocytes, co-cultured with MSCs for different times, were non-significant. The inhibitory effect of MSCs on B lymphocytes was temporary and reversible. Conclusion MSCs have immunoregulatory effects on B lymphocytes, and its mechanisms are complex, not only correlating with the concentration of MSCs but also the action between cells and the secretory cytokine of MSCs.