Objective To study the possible mechanisms of aprotinin in the protection of platelets. Methods Cytosolic free calcium concentration ([Ca 2+ ]i) was determined in calcium fluorescent indicator Fura 2/AM loaded washed human platelets by using dual wavelength spectrofluorophotometer. The mean value of resting [Ca 2+ ]i and the changes of [Ca 2+ ]i response to thrombin and aprotinin were observed. Results In the presence of extracellular Ca 2+ at the dose of 1 mmol/L, the resting level of [Ca 2+ ]i in platelets of human was (151.840?28.719) nmol/L. Thrombin stimulated the rise in [Ca 2+ ]i in the presence of Ca 2+ at the dose of 1 mmol/L, and the effects were inhibited by aprotinin in a concentration dependent manner ( P

BACKGROUND: Besides being a basic growth factor crucial to maintain and promote the development, differentiation and survival of the central nervous system, nerve growth factor(NGF) also plays an important role in the repair of injured peripheral nerves.OBJECTIVE: To investigate the effect of the muscular injection of NGF on the regeneration and functional recovery of rat sciatic nerve after crush injury.DESIGN: A randomized controlled pilot study in rats with repeated observation and measurement.SETTING: Center for new drug evaluation in a military medical university.MATERIALS: This study was performed in the Center for New Drug Evaluation, Department of Basic Medicine, Second Military Medical University during the period from July 1999 to March 2000, using 40 SD rats weighing 200 to 250 g(of either sex of half number) provided by the Sino-British SIPPR/BK Lab Aninal Ltd (Shanghai).METHODS: Forty rats were randomized into high-, mid- and low-dose NGF treatment groups, normal control group and model control group. The sciatic nerves were clamped at 6 nm distal to the sciatic notch to induce a 4-mm-wide area of crush injury. In the high-, mid-; and low-dose NGF groups, the rats were given NGF at 8, 4 and 2 μg/kg per day(corresponding to 1.6 × 10 3, 8 × 10 2 and 4 × 10 2 IU/kg per day) respectively via the muscular injection for 56 consecutive days.(NCVs) and sciatic function index(SFI) at different time points after the RESULTS: Compared with that of the model control group, the NCVs significantly increased in the high-dose NGF group 35 and 56 days after the injury,and in the mid-dose NGFgroup at 35 days(t=2.32-5.14, P ＜0.05-0.01 ). The SFIs significantly increased in all NGF-treated groups at 14 days ( t = 2. 29-6.28, P ＜ 0.05-0.01 ), with the recovery most conspicuous in high-dose NGF group; No significant difference in the SFIs was found between the NGF-treated groups on the 56th day. Morphological examination of the tissues identified no significant difference in the nerve myelin sheaths and axons in NGF-treated groups as compared with the normal control group,while in the model control group, myelin sheath dislocation with unclear microstructure was observed, accompanied by Schwann cell degeneration and necrosis.CONCLUSION: NGF promotes the repair of the damaged nerve myelin sheath and axon and stimulates nerve fiber regeneration and function recovery of the crushed rat sciatic nerves.

Objective To investigate the effect of isoflurane on the spatial learning and memory in aged mice,and whether this is associated with the changes of Caspase-3 expression and apoptosis in brain.Methods Twenty-four CO57BL/6 aged mice(16 months)were randomly divided into isoflurane treatment group(Iso Group,n=12) and sham control group (Con Group,n=12).Mice in Iso group were exposed to 1% isoflurane in carrying gas of 30% oxygen,balance nitrogen in a warmed,humidified chamber for4 h per day for2 days.For Con group,animals were treated at the same condition with only carrying gas.After anesthetic exposures,behavioral testing was performed using the Morris water maze(MWM),and then changes of Caspase-3 expression and apoptosis in hippocampus CAI,dentate gyrus(DG) and cortex(CX) in brain were determined by using immunofluorecence staining and TUNEL staining.Results In hidden-platform training of MWM,the mean escape latency to platform showed no significant difference between the two groups (F=0.007,P=1.235),but the mice in Iso group showed obviously impaired retention of memory by spending more percentage of time swimming in the probe quadrant as compared to the control animals in the probe test((34.5±5.0)%vs(45.1±4.9)%.P<0.01).There was no significant difference in average swimming speed during the MWM testing trials between the two groups (F=1.537,P=0.241).A few Caspase-3 and apoptosis positive cells were found in hippocampus Cal,DG and CX regious,while no difference was found in the density of positive cells between the two groups(P>0.05).Conclusion 1% isoflurane repeatedly exposure significantly impaires the spatial reference memory in aged mouses,however does not significantly change the expression of easpase-3 and apoptosis in brain.

Ketoconazole(KET) is a new imi-dazole derivative with broad antimycotic spectrum. In order to verify the clinical toxic and side effect and its properties in animals, we made a long-term toxicity test for 30 days. Dosages of 70, 35 and 17. 5 mg?kg-1?d-1(e-quivalent to 21, 10. 5 and 5. 2 times of the clinical dosage) were given ig to dogs. The salivation , vomiting, anorexia, decrease in heart rate and loss of weight occurred in the large dosage group. Half of the dogs died from toxicosis within ig 15 days. Laboratory examination showed that the activities of ALT, LDH and ALP, the content of T-BIL, BUN in serum in-creased in this group. Pathological examination revealed that there were some pathological changes in the liver, kidneys, adrenal glands and sex gland in the group. There were no significant changes in other dosage groups compared with the normal control group. After withdrawal of KET, all toxic symptoms disappeared and the abnormal indexes were restored. The results indicated that toxic target organs of KET were liver, kidney, adrenal gland and sex glands. The safe dosage for dogs was about 17. 5 mg?kg-1?d-1.

Objective To analyze the rules for acupoint selection of warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis based on data mining technology. Methods CNKI, Wanfang Data, CBM, VIP, PubMed and Cochrane Library were searched for relevant literature about warming acupuncture and moxibustion in treatment of knee osteoarthritis. Database was built according to the search results. Association rules were used to analyze the rules for acupoint selection. Results Totally 189 papers were included, involving 189 prescriptions of acupuncture and moxibustion, 46 acupoints, 1270 times of acupoint selection. There were 1119 times of neighboring acupoint selection (88.1%), 151 times of distant acupoint selection (11.9%), 654 times of acupoint selection in yang meridian (51.5%) and 283 times of acupoint selection in yin meridian (22.3%). Conclusion Warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis focuses on acupoints in Stomach Meridian of Foot-Yangming, Spleen Meridian of Foot-Taiyin; the neighboring acupoint selection is the main method, combined with evidence-based distant acupoint selection; acupoint selection from yang meridian was emphasized.

Objective:To investigate the long-term toxicity of recombinant human interleukin-11(rhIL-11) in cynomolgus. Methods: Eighteen cynomolgus were randomized into 4 groups: control group(2/sex), low dose group(2/sex), medium dose group(2/sex)， and high dose group(3/sex). The drug groups were sc adminstered 0.1, 0.3 and 1.0 mg/kg of rhIL-11 for 90 days with a 30-day recovery period. The clinical signs were observed, electrocardiogram, hematological, biochemical, urinary and immunological parameters were measured, organ masses were weighed, bone marrow and pathological histology were observed. Results: The food consumption, body mass of the drug groups were decreased, the body temperature was increased transiently. One of the low dose group showed restricted movements and tremors. One of the high dose group vomited and another died. Reduced red blood cell(RBC) count, hemoglobin(Hb) concentration, hematocrit(Hct), mean corpuscular volume(MCV), mean corpuscular hemoglobin(MCH), and mean corpuscular hemoglobin concentration(MCHC), dose-related increase of platelet(Plat) counts were present in drug groups. Biochemical examinations revealed dose-related decreases in serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH), total proteins(TP) and albumin(Alb) increases in serum alkaline phosphatase(ALP) levels. Positive antibody responses were seen and circulatory immune complex(CIC) was significantly increased in all drug groups. Hypertropy of marrow megakaryocyocytes was noted in the medium and high dose groups. The heart and liver masses were slightly increased in all treatment groups. Treatment-related microscopic findings included dose-related degeneration in the liver and the kidney. The adverse effects were reversed by the end of the recovery period. Conclusion: The target organs and systems are blood, liver, kidney, immmue system and bone marrow. The toxicity injuries were reversible and the no-toxic-effect level is 0.1 mg/kg.

Objective To investigate the expressions of CDX2 and β-catenin in acute lymphoblastic leukemia (ALL) and their correlation. Methods Real-time PCR was used to determine the expressions of CDX2 and β-catenin mRNA in 43 de novo ALL and 30 non-malignant patients (used as control). Results The positivity rate of CDX2 mRNA expression in ALL group was 93%, but CDX2 mRNA expression couldn′t be detected in the control group (P<0.01). The mRNA expression of β-catenin could be detected in patients in both two groups, butβ-catenin mRNA expression in the ALL group was significantly higher than in the control group (P < 0.01). And mRNA expressions of CDX2 andβ-catenin were significantly correlated with WBC counts and LDH level (P<0.01). When the ALL patients acquired complete remission (CR), the mRNA expressions of CDX2 and β-catenin were significantly decreased compared with their newly diagnosed status , while disease-relapsed the mRNA expressions of CDX2 andβ-catenin were increased again. There was significantly positive correlation between CDX2 and β-catenin mRNA expressions (r = 0.835, P = 0.000). Conclusion Up-regulation of CDX2 and activation of Wnt/β-catenin pathway coexist in the ALL patients and the mRNA expressions of CDX2 and β-catenin are positively correlated.

Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.

Objective:In order to enhance the early dignosis level and strengthen the understanding of pathogeny、pathology by stage and prognosis of primary gallbladder car-cinoma(PGC). Methods : The results of 679 patients of PGC from 1956 to 1998 in first affiliated hospital of Xi'an medical university were retrospectively analyzed. Results: Most of the patients belong to advanced stage when dignosised. The PGC were usually found among the old women patients. The sex ratio(female/male) was 3:1. There is a intimate relatioship between gallstone and PGC. About sixty percents of PGC might be found gallstone.The dignostic accordance rate was low between preoperation and postopera-tion.Most patients with PGC were found unexpectedly in operation due to gallstone or a-cute cholecystitis. To worse more, many of patiens with PGC were missed the opportunities of dignosis and treatment because of only paying attention to gallstone. Most of the pathological types were adenocarcinoma. The development of imaging examination is hoped to find early stage cases and improve its prognosis. Conclusion : It is a basic way for early dignosis and improving prognosis to strengthen the understanding of pathogeny, pathology by stage and prognosis and rationalize application of all kinds of exzamination means in PGC.

Objective To investigate the clinical effect of human neural stem cells transplantation on severe visual disability infants after cerebral palsy. Methods Cells obtained from the forebrain of an 11-week-old abortive fetus were cultured and expanded for 15 days, then injected into cerebral ventricle of 7 patients. Results Their vision of 4 patients improved, as well as changes of flash visual evoked potential and functional magnetic resonance imaging in a few days after transplantation. Conclusion Neural stem cells transplantation may benefit in some CP children with severe visual disability.