Objective To study the morphological character of the high endothelial venules(HEVs) and the expression of lymphocyte function-associated antigen-1(LFA-1),platelet endothelium cell adhesion molecular-1(PECAM-1) in the rat mesenteric lymph node,inquire into their morphological representation when lymphocytes pass through the HEVs and the regulative action of LFA-l and PECAM-1. Methods To observe their morphological representation when lymphocyte pass through the HEVs wall and analyze the expression of LFA-1,PECAM-1 with the light microscope,electron microscope and immunohistochemistry method in the rat mesenteric lymph node. Results Lymphocytes contacted the endothelial cells by protuberances or membrane protuberances like welding spot,lymphocytes located in and between the endothelial cells or in the junction space of them in HEVs,there were lymphocytes in the lucid layer between inner and outer layers of basement membranes;LFA-1 mainly expressed at lymphocytes that adhered to endothelial cell in HEVs;PECAM-1 mainly expressed at the endothelial cells and the migrating lymphocytes.Conclusion 1.Lymphocytes contacted the endothelial cells by protuberances,then they got across the endothelial cells into the cellular junction space by the intraendothelial and interendothelial,and at last they emigrated from the basement membranes into the lymph tissue;2.LFA-1 involved in firm adheresion between lymphocytes and the endothelial cells;3.PECAM-1 probably had a relationship with that lymphocytes migrated endothelial cells.

In the background of the crisis of doctor-patient relationship and the effectiveness of medical humanities education doubts we need to think about how to achieve,transform and reflect the value of doctors' understanding on their professional meaning,how to realize the functional significance of doctorpatient cooperation and how to make doctors put themselves in patients' place by the curriculum and its operating mechanism.This paper discussed the external environment and the microcosmic environment from the externality and connotation of the humanities curriculum.And the way to reflect and achieve the value transformation of the medical humanistic curriculum in the mechanism of education was considered.This paper put forward combining the institutional mechanism of policy leading,the subject mechanism that is the basis of the curriculum,the constitutive mechanism of curriculum system and the cultural mechanism to make students study and internalize,and made some suggestions.

Objective:To observe the curative effect and security of nicardipine combined with metoprolol succinate sustained-release tablet on renal hypertension.Method:65 renal hypertension patients were randomly divided into two groups.The patients in the nicardipine group were treated with nicardipine 80 mg q8h,while the patients in the therapy group were treated with nicardipine 40mg qd and metoprolol succinate sustained-release tablet 47.5-95mg qd for 8 weeks.Their blood pressure,heart rate,hepatic function,renal function,blood glucose and lipid were observed and recorded before and after the treatment.Result:The total efficiency was 74.4%in the controlled group and 88.5%in the therapy group,respectively. There were no obvious changes or obvious adverse reactions in their heart rate,hepatic function,renal function,blood glucose and lipid after the treatment.Conclusion:Nicardipine combined with metoprolol succinate sustained-release tablet is efficient and safe but more efficient than nicardipine alone in the treatment of renal hypertension.

AIM: To establish an analytical method for detecting 14 kinds of amino acid in Wuling Capsule(xylaria) by HPLC-FLD. METHODS: After being derivated in precolumn with o-phthalaldehyde and ?-mercaptoethanol,the amino acids were eluted by gradient elution and were detected with a fluorescence detector (Ex = 340 nm,Em = 450 nm). The mobile phase consisted of 0. 1 mol/L NaH2PO4 ( adjusted to pH 6. 8 by phosphoric acid) and methanol with a flow rate of 1. 0 mL/min,column temperature was set at 30 ℃. RESULTS: The deriva-tives of 14 amino acids with o-phthalaldehyde could be separated with a stable base line within 40 min. The linear range of the detection was 5-200 nmol/mL and correlation coefficients were over 0. 999 0. CONCLUSION: The method is simple,accurate and well reproduced for the determination of amino acids in Wuling Capsule.

This paper introduces the rapid & mobile first-aid unit from brigade or regiment medical aid station,especially discusses its equipment,performance,personnel structure,and working features.The rapid & mobile first-aid unit can perform emergency operation and anti-shock treatment for the serious wounded in frontier at any moment.It meets the requirements in such aspects as fast & effective first-aid,mobile & flexible treatment,high cure rate and good practicality.

Objective To express the major allergen of Blattella germanica (Bla g 2) in Pichia pastoris and obtain the soluble protein. Methods The known Bla g 2 gene was used to design the primers which had the restriction enzyme sites. PCR method was applied to obtain the Bla g 2 gene. The gene fragment was then cut and ligated with the Pichia expression vector pGAPZaA, resulting in a recombinant plasmid pGAPZaA-Bla g 2. The linearized pGAPZaA-Bla g 2 was transformed into Pichia pastoris GS115 through electroporation, then screened to positive transformants, and the protein was expressed in YPD medium. Purification of the recombinant protein was achieved by metal (Ni2+) chelating affinity chromatography and Western-blotting assay indicated its IgE binding capacity. Results With the expressed reeombinanl protein, SDS-PAGE showed the presence of the product in the supernatant of the culture with Mr 45 000. After 3 days culture, the recombinant protein occupied 50% of the total proteins in the supernatant. The recombinant protein was purified and Western-blot demonstrated an adequate IgE binding capacity of the product. Conclusion A recombinant protein of Bla g 2 has been obtained, which is soluble in the supernatant and therefore can avoid a process of denaturalization and renaturation of the recombinant.

BACKGROUND: Osteosarcoma cell OS-9607 is a cell line cultured from human osteosarcoma tissue. To construct cDNA library of this cell line so as to clone target gene from it.OBJECTIVE: To construct cDNA library of osteosarcoma cell OS-9607 by switching mechanism at 5' end of RNA transcript (SMART) technique and analyze its quality.DESIGN: Verified experimental study SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA in May 2005. Osteosarcoma cell OS-9607 was cultured in CO2 incubator. Cells were pre-treated with DM for 30 minutes, recovered for 24 hours in standard culture medium. The whole RNA of osteosarcoma cell OS-9607 was isolated by RNAease reagent kit. The quantity and integrity of total RNA was detected by ultraviolet spectrometer and electrophoresis on a denaturing formaldehyde agarose gel stained by ethidium bromide (EtBr). METHODS: Total RNA was extracted from human osteosarcoma cell OS9607 and mRNA was isolated, cDNA of OS-9607 cells was acquired by in situ polymerase chain reaction (PCR) and long-distance PCR (LD-PCR),then the cDNA was ligated with dephosphorylated λ phage vector. The recombinants were constructed with SMART cDNA library construction kit.The size of cDNA inserts and the diversity of library were detected by PCR.MAIN OUTCOME MEASURES: The quality and integrity of total RNA, the titer of un-amplified and amplified library and recombination fraction; analysis of cDNA length of library RESULTS: ①The concentration of the isolated total RNA was determined and the measuring absorbance was between 260 nm and 280 nm. Its absorptance was about 1.9 A. 0.8％ denaturing formaldehyde agarose gel electrophorosis showed clear 28S rRNA and 18S rRNA,and the brightness rate of 28S rRNA and 18S rRNA (28S/18S) was 2:1. ②After linked to λ phage in vitro, packed and transfected host bacteria, the titer of unamplified library was 2.2×107 pfu/mL, and the titer of amplified 1 ibrary arrived to 4.5×1010 pfu/mL. The recombination fraction analysis showed that there were 8 blue plaques and 1 360 colorless plaques in one plate, so the recombination fraction was 99.5％. ③ The obtained cDNA library consisted of 1.6×106 recombinant bacteriophages and the average exogenous inserts of the recombinants was 1.5 kb.CONCLUSION: These results suggest that the obtained osteosarcoma cell OS-9607 cDNA library has high quality, which sets up a firm foundation for the further research on osteosarcoma cell OS-9607 and its elated genes screen.

Objective To study the expression-mode and dynamic transmutation of high mobility group box-1 (HMGB1) in hepatocytes of the mouse model with acute hepatic failure and to study the interaction beween HMGB1 and tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β). Methods The mouse model of acute hepatic failure was established by injecting D-galactosamine (D-GalN) and lipopolysaccharide(LPS). Immunohistochemistry SABC method was used to detect the HMGB1 expression at 6 time points. The enzyme linked immunosorbent assay (ELISA) was used to determine serum TNF-α. IL-1β levels. Paired t test was used for statistical analysis. Results The HMGB1 expression was detectable at 2 hours after injection, which dramatically increased over time and peaked at 24 hours after injection. The serum TNF-a level and IL-1β level increased right after injection. The TNF-a level peaked at 8 hours after injection with a maximum value of (473.42±22. 99) pg/mL. The IL-1β level peaked at 2 hours after injection with a maximum value of (724. 49±34. 24) pg/mL. Both cytokine levels slowly decreased after peaking. IL-1β level returned normal with (51. 49±36. 87) pg/mL. Conclusions HMGB1 is one of the most important factors during the development of acute hepatic failure, which can promote the secretion of TNF-α and IL-1β at early stage and be abundantly expressed under the effect of these cytokines at middle and late stages with the result of liver damage. This process is directly correlated with the development and severity of hepatic failure.

Injury of choledocho-pancreatico-duodenal junction refers to the penetrating injury of the bile duct, pancrea-tic duct or duodenal wall in the region of ampulla of Vater. It is often caused by improper operation of surgical instruments, and may lead to leakage of bile, pancreatic or duodenal contents into retroperitoneal space and chemical corrosion in a wide range of retroperitoneal soft tissue, which result in severe secondary infection or even death. Leakage of contrast media, hypertrophy of tissue and anatomical changes were the evidences for injury of choledocho-pancreatico-duodenal junction. Injury of choledocho-pancreatico-duodenal junction can be. divided into 4 types, and treatment selected according to different types of injury is neces-sary for the prognosis of patients.