BACKGROUND:As the bone and marrow tissue have very special structure, it is difficult to simultaneously display the bone with tough hard tissue and bone marrow tissues containing various immature hematopoietic cel s in the conventional process of pathological section preparation. OBJECTIVE:To choose the best decalcifying solution that cannot only completely remove the calcium in the bone tissue but also protect the structure of bone marrow tissues and cel s from damage. METHODS:Bone marrow tissues from the long bone of dogs were randomly divided into four groups. Under the same conditions, the bone marrow tissues were decalcified with 14%formaldehyde saline solution of nitric acid (group A), 14%nitric acid solution (group B), 20%A saline solution of hydrochloric acid formaldehyde (group C) and 20%A hydrochloric acid aqueous solution (group D). Decalcified time was recorded, fol owed by routine dehydration, section, hematoxylin-eosin staining and microscopic observation. Pathological section quality and hematoxylin-eosin staining were compared among the four groups. RESULTS AND CONCLUSION:Group A had the best sections and hematoxylin-eosin staining, strongest decalcified ability, shortest decalcified time and minimum damage to the bone marrow. Group B had the worst results of section and hematoxylin-eosin staining, in which, the bone tissues were loose and became yel ow and the bone marrow tissue were damaged greatly, and the decalcified effect was worse. Group C was worse than group A in decalcified ability, damage degree, section quality and hematoxylin-eosin staining results. Group D also had a better result of section and hematoxylin-eosin staining as wel as exhibited uniform decalcification effect and less damage to the bone marrow, which was ranked between group B and group C. Al the four kinds of decalcifying solutions have a good decalcification ability, but the section quality and hematoxylin-eosin staining results rank as fol ows:Group A>Group C>Group D>Group B. Taken together, 14%formaldehyde saline solution of nitric acid is ideal for the clinical preparation of pathological sections.

The in-vitro fertilization and embryo transfer technique has been widely applied in human insemination. The rate of successful insemination is gradual y rising, and the in-vitro fertilization directly determine the insemination outcome.
OBJECTIVE:To evaluate the difference between the two common using insemination methods, microdrop and open, in in-vitro fertilization and embryo development.
METHODS:A randomized study was conducted to compare microdrop and open insemination methods among non-male factor patients undergoing in-vitro fertilization and embryo transfer. A total of 1 175 cases were enrol ed in the research. There were 573 cases in the microdrop group, and 602 cases in open insemination group. The fertilization rate and embryo development in the two groups were compared.
RESULTS AND CONCLUSION:The fertilization failure rate [total fertilization failure rate+low fertilization rate (<25%oocytes fertilized)] in the microdrop insemination group was higher than in the open insemination group (11.9%, 3.3%, P<0.001), while the good quality embryo rate and pregnancy rate did not differ significantly between the two groups (al P>0.05). The open insemination method is a simple insemination method with a lower fertilization failure rate. As the fertilization is a highly complicated process involving many extrinsic and intrinsic factors, further study is needed to confirm the effects of the two insemination methods on in-vitro fertilization outcome.

OBJECTIVE: To investigate the pattern and value of the expression of T-bet in nasopharyngeal-associated lymphatic tissue (NALT) of patients with or without allergic rhinitis (AR). METHOD: T-bet and CD4/T-bet expression in adenoids, tonsils and nasal mucosa was respectively detected with single stain and double stain of immunohistochemistry. RESULT: The positive ratio of T-bet expression in tonsils, adenoids and nasal mucosa between AR group and the control group had statistically different (P < 0.05). There was statistical difference of T-bet expression in tonsils from AR group in three different age groups (P < 0.05). The difference was that T-bet expression in adults group was significantly lower than that in children group and adolescent group. There was no difference of T-bet expression among different age groups in non-AR group. There were some positive expression of CD4 and T-bet in tonsils, adenoids and nasal mucosa from two groups and most positive expression of T-bet on CD4 positive cell. CONCLUSION T-bet expression was down-regulated in NALT of patients with AR. T-bet expression of NALT is associated with allergic mucosal inflammation and functional status of NALT, as well as the weak Th1 response at the level of transfer factor in local mucosa of respiratory tract in AR patients.
Adolescent ; Adult ; Child ; Humans ; Lymphoid Tissue ; metabolism ; Palatine Tonsil ; metabolism ; Rhinitis, Allergic, Perennial ; metabolism ; T-Box Domain Proteins ; metabolism ; Young Adult

Objective To study the expressions of Nodal in normal gallbladder,and gallbladders with cholelithiasis,cholecystitis and carcinoma;and to study the impact of inhibiting or promoting Nodal expressions in gallbladder carcinoma on the Smad2/4 pathway and epithelial-mesenchymal transition.Methods Immunohistochemistry was used to detect the expressions and distributions of Nodal protein in 30 normal gallbladders,96 simple cholecystitis/calculous cholecystitis specimens and 42 gallbladder carcinoma specimens.The mRNA and protein expressions of Nodal in normal and malignant gallbladdcr mucosal epithelium cells and breast cancer cells were detected by RT-PCR,western blotting and wound healing tests.The impact of activating agents and inhibitors on the expression levels of Nodal and its signaling pathway Smad2/4 and EMT-related proteins were analyzed.Results Immunohistochemistry showed that the positive rates of Nodal in the gallbladder cancer group was significantly higher than that in the gallbladder stone group and the normal gallbladder group.The results were 83.3％ (35/42),44.8％ (43/96),6.7％ (2/30) respectively (P＜ 0.01).RT-PCR and Western blotting showed the expressions of Nodal in gallbladder carcinoma cells were higher than normal gallbladder cells (P＜0.05).After using rhNodal to up regulate the Nodal expression,the Smad2 protein phosphorylation was promoted and the EMT associated proteins were up-regulated.After using the inhibitor SB431542 to suppress the Nodal expression,the Smad2 protein phosphorylation decreased and the EMT associated proteins were down-regulated.Conclusions The expression of Nodal was closely related to cell proliferation and metastasis in gallbladder carcinoma.Tumor progression was promoted via the smad2/4 pathway through epithelial-mesenchymal transition.