AIM:To study chromosome aberration due to ethidium bromide (EB), a heterocyclic organic compound and an organic fluorescence dye commonly used in biochemical experiment, and to help further understanding the molecular mechanism of tumor or cancer induced by EB and other heterocyclic organic compounds. METHODS: The toxicity action of EB was evaluated from three aspects including DNA, chromosome and embryo stem cells (ESCs) using atomic force microscopy (AFM), and thereinto, the morphology structural difference of ESCs treated with two EB doses was also valuated. RESULTS: The morphological structures of DNA, chromosome and ESCs were dramatically damaged. The average height of DNA decreased 0.5 nm; chromosomal arms were ruptured from centromere location; molecules of cellular membrane congregated and loop-like structure formed, and ES cell masses were collapsed and became dead after large EB doses treatment and mesh-like morphological structure was discernable. CONCLUSION: The toxicity action of EB is strong and destroys the surface structure of DNA and chromosome. EB induces structural aberration of ES cellular membrane and cell death. The results indicate that the action of EB is externalized at gene level and cell level, which is important to study the carcinogenicity of EB.

Normal bacteria purification method was used in this study. Pseudomonas trivialis and Neurospora te-trasperma were screened from residues of traditional Chinese medicine (TCM). The fermentation conditions and flocculating function were also studied. The results showed that the selected strains grow well in the residues of TCM and their products can flocculate kaolin suspension solution. Based on these, we explored effects of various factors on the flocculated results in order to reveal the best fermentation condition among different combinations.

Scanning near-field optical microscopy (SNOM) is developing on the base of near-field optical theory and SPM technique, and producing the highest optical resolution to break through the diffraction limit. It is able to study cellular ultrastructure, function and interaction between biomolecules without destructing the living cells. Recent advances in applying SNOM to cellular and molecular biology were introduced in this review.

Objective To explore the association between rs3758539G-803A and rs10882283 T-179G polymorphism of retinol binding protein 4 (RBP4) and rosiglitazone response in Chinese type 2 diabetes mellitus (T2DM) patients.Methods A total of 472 Chinese T2DM patients and 198 healthy subjects were enrolled to identify G-803A and T-179G genotypes using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP ).assay.Forty-two T2DM patients with different G-803A or T-179G genotypes were selected to undergo a 12-week rosiglitazone treatment (4 mg/d).Serum fasting plasma glucose (FPG),postprandial plasma glucose (PPG),fasting serum insulin (FINS),glycated hemoglobin (HbAlc),postprandial serum insulin ( PINS),triglyceride (TG),low-density lipoprotein-cholesterol ( LDL-c),and high-density lipoprotein-cholesterol (HDL-c) were determined before and after the rosiglitazone treatment.Results T2DM patients with RBP4 G-803A GG genotype showed lower TG and LDL-c concentrations compared with that in the GA +AA genotype subjects.T2DM patients with RBP4 T-179G TT genotype showed lower waist-to-hip ratio (WHR),FPG and FINS values compared with that in the TG + GG genotype individuals.Patients with GG genotype of RBP4 G-803A had an enhanced rosiglitazone efficacy on FPG and FINS compared with that in the GA + AA genotype group.Patients with RBP4 T179G TG + GG genotype showed an enhanced rosiglitazone efficacy on HbAlc level compared with that in the TT genotype group.Conclusion RBP4 G-803A and T-179G polymorphism might be associated with the development of T2DM and affect the therapeutic efficacy of rosignitazone in Chinese T2DM patients.

AIM: To study the cytoskeleton of mesench ymal stem cells (MSCs), the ultrastructure and function relationship by using atomic force microscope (AFM). METHODS: The ultrastructures and morphological feature of MSCs c ultured for 1 d and 5 d were studied by AFM. RESULTS: The special structures that possess peculiar morphologi cal characteristic of MSCs such as cytoskeleton, pseudopod, microfilament etc we re identified by AFM, and these special structures are difficult to observe unde r electronic microscopy or other conventional optical microscopy. CONCLUSIONS: AFM is a powerful tool to study ultrastructures, mo rphological features, and cytoskeleton of stem cells in near physiological condi t ions. Its application prospect in cellular biology is extensive. The special cyt oskeleton and other structures of MSCs observed above may represent the structur al base of multi-differentiation potential of MSCs.

Atomic force microscope (AFM) was used to study biotoxicity of food preservative sodium benzoate (SB) at the single cellular level. Lymphocyte morphology and membrane ultrastructure treated with SB at different concentrations and time were analyzed visually. As compared to the normal lymphocyte, the cell morphology and membrane was significantly changed and its ultrastructure was also complicated. After treated with SB, the Rp-v, Rq, Ra and Z values were changed. The statistical analysis of lymphocytes after treated with SB was studied, and discussed its mechanism.
Animals ; Cell Membrane ; ultrastructure ; Lymphocytes ; drug effects ; pathology ; Mice ; Mice, Inbred BALB C ; Microscopy, Atomic Force ; Sodium Benzoate ; toxicity

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.

Objective To investigate the clinical manifestations,imaging features and prognosis of mixed liver cancer.Methods The clinical and pathological data of 26 patients with combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CC) pathologically diagnosed after liver resection were retrospectively analyzed,and the relevant factors affecting the prognosis were statistically analyzed.Results Of the 26 cHCC-CC patients,19 were on background of chronic viral hepatitis,including 17 chronic viral hepatitis B and 2 chronic viral hepatitis C.There were 18 cases having AFP≥20ng/ml,9 cases were with carbohydrate antigen 199 (CA199) ≥ 37U/ml,and 5 cases with carcinoembryonic antigen (CEA) ≥5ng/ml.The 1-,3-,and 5-year postoperative survival rates were 68.8％,34.4％,and 17.4％.Multivariate analysis showed that serum CA199≥37U/ ml(x2 =5.687,P =0.019) was an independent risk factor for patients' survival.Conclusion Most cHCC-CC is found in association with chronic viral hepatitis.Serum CA199 ≥37U/ml is an independent risk factor affecting patients survival.

Objective:To evaluate the clinical efficacy and changes between Dynesys and Posterior lumbar interbody fusion (PLIF) in the treatment of two-level lumbar degenerative disease.Methods:43 consecutive patients with lumbar degenerative disease were treated using the Dynesys or PLIF between June 2010 and June 2012. In all patients, 23patients were implanted Dynesys and other patients for PLIF. The follow-up period was at least 60 months. Used the Visual analogue scale (VAS) and Oswestry disability index (ODI) to evaluate the clinical outcomes. And during the follow-up period, collected the data for the intervertebral height and the range of motion (ROM), for the operation section and the upper adjacent vertebral. MRI and Grading Scale from the University of California at Los Angeles (UCLA) were used to define the change of intervertebral disc signal.Results:The ODI index and VAS score both improved significantly at the final follow-up evaluation ( P<0.05), as compared to the basal line values. And in terms of imaging, there were no significant difference in the vertebral height of the operation section and the upper adjacent vertebral. The range of motion (ROM) for the operative section in the group of PLIF was significantly lower than that in Dynesys group ( P<0.05). And when it comes to the upper adjacent vertebral space, it was higher at the same time ( P<0.05). According to the UCLA Grading Scale, there were 3 cases in the Dynesys group and 11 in the PLIF group had radiological adjacent degeneration, the difference was significant between two groups ( P<0.05). And when it comes to the second intension, there were 3 patients in PLIF groups (TLIF 1; PLIF 2) but only 1 in Dynesys groups. Conclusion:Dynesys and PLIF are both effective for lumbar degenerative disease and show good medium and long-term clinical and radiographic results. But when it comes the risks in developing ASD and the ROM of adjacent segments, Dynesys stabilization can be the better choice.

Background Bromadiolone is the second-generation anticoagulant rodenticide widely used all over the world. Exposure to bromadiolone in early life stage can lead to neurodevelopmental toxicity, but its toxic mechanism of neurodevelopment is not clear so far. Objective To investigate the developmental neurotoxicity and mechanism of bromadiolone to zebrafish embryos. Methods Zebrafish embryos were randomly divided into four groups: a solvent control group (dimethylsulphoxide) and three bromadiolone exposure groups (0.39, 0.78, and 1.18 mg·L⁻¹). The exposure period was from 4 h to 120 h post-fertilization. The number of spontaneous movement per minute was recorded at 24 h post-treatment. The locomotor ability of zebrafish larvae and the activity of acetylcholinesterase (AChE) were tested at 120 h post-treatment. The relative expression levels of neurodevelopment-related genes (elavl3, gap43, mbp, and syn2a) were measured by fluorescence quantitative PCR. Results Compared with the control group, the number of spontaneous movement per minute at 24 h decreased significantly in the 1.18 mg·L⁻¹ bromadiolone exposure group (P<0.05). Compared with the control group, the total distance travelled of the zebrafish larvae in the 0.78 and 1.18 mg·L⁻¹ bromadiolone exposure groups decreased by 60% and 69% respectively (P<0.05, P<0.01), and the total movement time decreased by 34% and 65% respectively (P<0.05, P<0.01). The AChE activity in the 1.18 mg·L⁻¹ bromadiolone exposure group increased by 36% when compared with the control group (P<0.05). The fluorescence quantitative PCR results showed that compared with the control group, the expression levels of neurodevelopment-related genes elavl3, syn2a, and mbp were significantly down-regulated by 66%, 69%, and 65% in the 1.18 mg·L⁻¹ bromadiolone exposure group respectively (P<0.01), the expression level of gap43 was up-regulated by 56% in the 0.78 mg·L⁻¹ bromadiolone exposure group (P<0.01) and down-regulated by 34% in the 1.18 mg·L⁻¹ bromadiolone exposure group (P<0.05). Conclusion Bromadiolone exposure could inhibit spontaneous movement and locomotive behavior, down-regulate the expression levels of neurodevelopment-related genes, hinder the release of neurotransmitters, and result in neurodevelopmental toxicity in the early-staged zebrafish.