Abstract:Objective: To demonstrate whether some important inflammatory markers rise in degenerative cervical intervertebral disc, and to investigate pathogenesis of cervical spondylosis in biochemistry. Methods: In this experiment,20 white New Zealand rabbits were used and divided into 2 groups: control group and model group. Each group consisted of 2 subgroup: 3 month group and 8 month group. The experimental model of degenerative cervical disc in the rabbit was established by resection of the cervical supraspinous and interspinous ligaments and detachment of the posterior paravertebral muscles from the cervical vertebrae. Then animals in each group were killed 3 months and 8 months after operation, respectively. The concentrations of histamine,5-OH-tryptamine,PGE2 and 6-keto-PGF1α were assayed in all animal disc samples. Results: The contents of 4 inflammatory mediators from the discs of model groups were overtly higher than that of control groups, respectively. The contents of 4 inflammatory mediators had no statistical difference between model groups of 3 month and 8 month. Conclusion: The degenerated cervical disc is inflammatory, and the inflammatory mediators produced by the degenerated cervical intervertebral disc may be directly related to the pathogenesis of cervical spondylosis

Abstract:Objective: To express recombinant human IL-12 which has the activity of stimulating the proliferation of T cells in vitro by baculovirus/insect cell system. Methods: Insert p35 and p40 cDNA into baculovirus transfer vector pAcUW42, then co-transfect insect Sf9 cells with AcUW1.lacZ baculobvirus DNA for constructing IL-12 expression recombinant baculovirus by homologous recombination. Results: Culture supernatant of Sf9 cells infected by IL-12 recombinant baculovirus which selected by a series of assay of baculovirus plaques, ELISA and Western Blotting, could stimulate the proliferation of T cells from normal human peripheral blood as standard IL-12 did. Conclusion: We have established the IL-12 baculovirus/insect cell expression system whose product has the activity of stimulating the proliferation of T cells in vitro

Abstract:Objective: To construct pMCLacⅠ/Neo plasmid as an effective vector for studying the different mechanisms of mutation between expressed and non-expressed genes in mammalian cells. Methods and Results: A pMCLacⅠ/Neo plasmid, containing 2 copies of lacⅠ gene as the mutation target genes, was designed and constructed by common molecular cloning techniques. In this plasmid, only one of the lacⅠ genes was under the control of CMV promoter and could be transcribed in human and mammalian cells. After structure identification by restriction analysis, pMCLacⅠ/Neo plasmid was further studied with its function. It was first introduced into E.coli cells to test the function of the 2 lacⅠ target genes. The results indicated that both lacⅠ genes could regulate the expression of lacZα reporter gene in DH5α host cells. Whether the 2 target genes could imitate the functional states of expressed and non-expressed genes in mammalian cells was also analyzed. A NIH3T3 cell line containing copies of a stably integrated pMCLacⅠ/Neo plasmid was established through liposome-mediated transfection, and the functional states of the 2 lacⅠ target genes in the cell line were analyzed with RT-PCR. The results showed that only one of the 2 lacⅠ genes could be transcribed in the NIH3T3 cells. Conclusion: A new plasmid as a vector for studying gene mutation, in which 2 lacⅠ target genes could imitate the functional states of expressed and non-expressed genes in human and mammalian cells respectively, is constructed.

Abstract:Objective: To amplify large number of dendritic cells (DCs) from murine bone marrow and assay their morphological, immunological characteristics. Methods: Lymphocytes, monocytes-macrophages and granulocytes of murine bone marrow were depleted using immuno-Minimagnetic cell sorter. The selected cells were cultured with granulocyte-macrophage colony-stimulating factor, and the floating cells were collected 8 d after culture. The phenotype of collected DCs was analyzed, and their morphology was observed under electron microscopy. Results: (2-5)×106 DCs were obtained from bone marrow of one mice, and the purity of DCs was more than 80%. The cells expressed specific antigens (DEC205) of DCs and had typical morphology of DCs. Conclusion: A method to generate large number of DCs from murine bone marrow is established, which contribute to further studying immunological functions of DCs.

Abstract:Objective:To study the apoptosis of bone marrow hematopoietic cells and Fas expression in 60Coγ irradiated mice. Methods: Mice were whole-body-irradiated with 4.0 Gy γ-rays.The bone marrow hematopoietic cells were analyzed by electron microscopy(EM),electrophoretic analysis of DNA and flow cytometry. Results:EM showed that there were nuclear contraction and chromatin condensation.Agarose gel electrophoresis of DNA showed ladder pattern.The peak percentage of apoptotic cells occurred 8 h after irradiation. Fas expression significantly increased in bone marrow hematopoietic cells than that of control group 4 h after irradiation. Conclusion: The results indicate that apoptosis occurs in bone marrow hematopoietic cells of mouse with γ-irradiation.The expression of Fas may play a role in raidation-induced apoptosis of hematopoietic cells.

Abstract:Objective: To observe the effect of endothelin receptor antagonist on blood flow and function of injuried spinal cord in rats and explore mechanisms of ET in spinal cord injury (SCI). Methods: The thoracic spinal cord of rats was compressed by a load of 35 g for 5 min, which caused moderate injury. The rats were assigned into 3 groups: control group, injuried group, PD142893 group. The changes of spinal cord blood flow (SCBF) were measured by way of Laser Doppler flowmetry during 8 hours and the neurological function of injuried rats was evaluated one week after spinal cord injury. Results: PD142893 significantly increased early SCBF at compression sites after SCI by intrathecal injection and partly restored the motor function of the damaged spinal cord. Conclusion: ET plays a key role in secondary injury after acute spinal cord injury and that intrathecal injection of ET receptor antagonist PD142893 can partly reverse the progression of SCI.

Abstract:Objective: To understand the effects of rectangle-shaped intramedullary nailing on the bone callus and the bone morphogenetic protein content in osteoblast. Methods: Twenty-four New Zealand rabbits tibial fractures were internally fixed by RIM and plates, animals were killed in groups successively at 14, 21, and 28 d and samples from bone callus of the fracture site were reacted with BMP-Ab by immunochemistry method. And the results were quantified by IAS. X-ray films were taken of both tibia 56 d after operation, and the X-ray films were also quantified by IAS. Results: BMP content in single cell from flexible fixation group is significantly higher than that of plating group, and bone callus content from the RIM fixation group is also richer than that of the plating group. Conclusion: After RIM internal fixation, there is micromovement between the bone fragments, and this may stimulates the secretion of BMP. And the increment of BMP secretion may lead to richer bone callus formation.

Abstract:Objective:To study the protecive effect of ultraprofound hypothermia on canine ischemic brain. Methods:Seven adult dogs underwent closed-chest extracorporeal circulation with both external and internal body cooling. As they were cooled, progressive hemodilution with Haemaccel was employed until the hematocrit decreased to 5%or so. Continuous circulation and a core temperature at 7-10℃ were maintained for 90 min. During rewarming, the animals were autotransfused and allowed to recover. Results:Of the 7 animals, one died intraoperatively due to technical factors related to closed-chest extracorporeal circulation, 6 surviving animals showed normal neurological functions, but died of dysfunction of lungs 2-4 h after resuscitation. Pathological examination revealed mild ischemic lesions of myopcardiac structure, pulmonary edema with microscopic alveolar hemorrhage and no histological abnormality of brain, liver or kidney. Conclusion:The canine brain, liver and kidney can tolerate ischemia for 90 min under ultraprofound hypothermia, despite the dysfunction of lungs after resuscitation.

Abstract:Objective:To investigate airway ciliary ultrastructure and mucociliary clearance(MCC) in COPD patients with heavy smoking. Methods:Bronchial biopsy from 10 patients and 6 healthy volunteers during fiberoptic bronchoscopy was performed and submitted to ultrastructural study of cilia.Using Emisson Computed Tomography of radioactive particles inhalation technique,MCC of 7 cases and 7 normal control subjects was evaluated. Rusults: The percentage of axonemal ultrastructure abnormalities(AUA) was markedly higher in patients group［(19.2±2.8)%］ than that in control group［(11.6±2.3)%,P<0.01］. The half time of radioactivity aerosol particles being cleared(MCC marker) in right lung center-region　of interesting（C-ROI） was significantly higher in patients group［(1943±629)s］than that in control group［(1207±276)s,P<0.05］.There was a significant　correlation between AUA and MCC (r=0.704 5,P<0.05). Conclusion: Heavy smoking and chronic airway infection cause significant increase in airway AUA percentage and decrease in MCC of respiratory tract, and Ciliary ultrastructure is coincided with mucociliary clearance of respiratory tract.

Abstract:Objective:To evaluate the diagnostic accuracy,complications, and usefulness of CT-guided transthoracic needle biopsy of small pulmonary nodules. Methods: Between August 1991 and May 1997,158 patients underwent transthoracic needle biopsy of pulmonary lesions.Thirty-five lesions were presented small nodules,defined as lesions of 2.0 cm or less maximum diameter.A total of 38 biopsies were performed in the 35 lesions. Results: There were 11 benign lesions and 24 cancers (19 primary， 5 metastatic).Transthoracic needle biopsy findings were positive for cancers in 21 cases.In all 3 false negative lesions, only one biopsy was performed.There were 11 true-negative results and no false-positive results. Positive and negative predictive value were 100％ and 79％，respectively. Pneumothorax occurred in 5(13％)of 38 biopsies,but no chest tube placement was necessary. Conclusion:CT-guided transpulmonary biopsy of small pulmonary nodules can produce diagnostic yields comparable with that of larger lesions, and has a definitive diagnostic value of clinical utility.

Abstract:Objective: To provide the basis for determining the suitable collecting time and processing methods of safflower. Methods: Based on the introduction and cultivation of safflower, the crude drug was collected in different flowering days, and dried in the shade, in the sun, at 45℃ and 60℃. The contents of the components in the crude drug were measured by UV and HPLC. Results: The contents of the components in safflower crude drug were significantly affected by collecting time (P < 0.01), while they were not affected by the processing methods. Conclusion: The suitable collecting time of safflower crude drug are the third flowering day. The quality of the crude drug dried bellow 60℃ is not affected.

Abstract:Objective: To study the influence of mannitol on the pharmacokinetics of fluconazole. Methods:After oral administration the concentration of fluconazole in serum was determined by HPLC. The concentration of fluconazole in serum-time profiles were conformed by 3P87 program. Results and conclusion: The pharmacokinetic parameters of fluconazole alone and combined with mannitol were: ka=(0.19±0.08) and (0.40±0.13) h-1 (P<0.05), t1/2ka =( 3.97±1.21) and (2.05±1.08) h (P<0.05). No significant differences were noted between fluconazole alone and in combination with mannitol in cmax and AUC (P>0.05).

Abstract:Objective: To discuss the therapeutic effect of immunosuppresant for adult polycystic kidney disease (APDK). Methods: Collecting 24 cases of APKD patients undergone the renal transplantation for 1-8 years (mean time is 3.18 years). The checking items were waist pain,haematuria,hypertension and polycystic kidney size before and after renal transplantation. Immunosuppressants include cyclosporine, azathioprine and prednisone. Results: After transplantation, release of waist pain was most obvious; occurrence rate of haematuria decreased from 83.3%to 12.5%(P<0.01); hypertention did not change in 1 month but improved in 1 year (P<0.01); polycystic kidney size examined by B-ultrasound showed no difference and stayed in “steady” state. Conclusion: Immunosuppressants contribute to all the above improvements. APKD may be gene-depended immunodisease or immunity plays an important role in its pathogenesis. So immunosuppressant may be therapeutically effective for APKD.

Abstract:Objective: To investigate inhibitory effect of recombinant TGFα-PE40 (TP40) on cultured vascular smooth muscle cells (SMC). Methods: Expression of epidermal growth factor receptor (EGFR) in SMC was analysed with immunohistochemistry. Inhibitory effects of TP40 on SMC proliferation and protein synthesis were analysed by crystal violet staining and 3H-leucine incorporation. Competition assays were performed by the addition of excess of EGF. Results: Expression of EGFR was found in rapidly proliferating SMC, not found in quiescent SMC. Inhibitory effects of TP40 on rapidly proliferating SMC proliferation and protein synthsis was much higher than on quiescent SMC. Excess EGF can completely block inhibitory effects of TP40. Conclusion: It indicates that TP40 can significantly inhibit the proliferation of rapidly proliferating SMC, but less confluence quiescent SMC. The cytotoxic effects of TP40 are specifically mediaed by EGFR.

Abstract:Objective: To investigate the relationship between age or period of hypertension and morphological remodeling of renal arterioles. Methods: Under dilated conditions, the morphometry of the renal arterioles were studied microscopically on kidney slices combining computer-assistant image analysis in 16, 26, 40 and 55 weeks of SHR. Results: Compared with age-matched Wistar-Kyoto, the wall thickness and ratio of wall thickness to inner diameter were much increased in the renal arterioles, while the wall area in the arterioles of 50 to 200 μm outer diameter increased. The inner diameter decreased in the arterioles of 20 to 50 μm outer diameter in 40 and 55 weeks of SHR. Hypertension and age were both related to the morphological remodeling of renal arterioles in SHR. Conclusion: Hypertension is the main reason of the remodeling of the renal arterioles in SHR from 16 to 55 weeks. The remodeling of the renal arterioles less than 50 μm outer diameter is due to the rearrangement of medial smooth muscle cells (SMC), but the remodeling of the renal arterioles more than 50 μm outer diameter is due to proliferation of the medial SMC.

Abstract:Objective: To assess the effect of high hepatic activity on the inferior myocardium in 99mTc-nitriles SPECT myocardial perfusion imaging. Methods :A heart and liver phantom was used. Images were performed in different heart-to-liver activity ratios(H/L) and heart-to-liver distances. The acquired projections were reconstructed with both contrast-enhancement(standard) and quantitative Ramp filters(Ramp-C and Ramp-Q). Considering the inferior-to-anterior (I/A) wall activity ratio in the no-liver phantom study reconstructed with Ramp-C as “normal”, the rate of decrease of I/A in other images was calculated. The relations between the rates of decrease of I/A and the various H/L and heart-to-liver distances were analyzed by linear correlation. Results: The decrease rate of I/A was linearly negatively correlated with the H/L(r=-0.986,P<0.01) and the heart-to-liver distances (r=-0.970,P<0.01).The I/A reconstructed with Ramp-Q were higher for 2.2%？10.0%than that with Ramp-C.Conclusion: High hepatic activity may create artifactual count loss in inferior wall in 99mTc-nitriles SPECT myocardial perfusion imaging.The degree of count loss is proportional to the activity level of a liver and inversely proportional to heart-to-liver distance.The extent of count loss, while tomographic images are reconstructed with Ramp-Q, is smaller than that with Ramp-C.

Abstract:Objective:To compare the effects of intermittent cold blood cardioplegin, continuous cold blood cardioplegin and continuous warm blood cardioplegin on the recovery of cardiac performance using the cat cardiopulmonary bypass, after longtime cardiac arrest. Methods:twenty-four cats weighting 3-3.5 kg were divided at random into 4 groups. Hearts in group Ⅰ were not subjected to ischemic arrest, hearts in group Ⅱprotected with intermittent cold blood cardioplegia, hearts in group Ⅲ with continuous cold blood cardioplegia, hearts in group Ⅳ with continuous warm blood cardioplegia. After 180 min cardiac arrest, all hearts were reperfused for 60 min. Results:the cardiac perfomance was significantly reduced in group Ⅱ and Ⅲ, especially in group Ⅱ protected with intermittent cold blood cardioplegia, and kept almost normal in group Ⅳ with continuous warm blood cardioplegia. LDH-1 and MDA content rose to a higher level in group Ⅱ and Ⅲ after reperfusion, but almost normal in group Ⅳ protected with continuous warm blood cardioplegia. Conclusion: Our data indicate that the method of continuous warm blood cardioplegia is safe and effective and represents a new conceptual approach to maintaining excellent myocardial preservation during cardiac arrest. It can enhance the recovery of cardiac performance after reperfusion

Abstract:Objective:To observe the kinetics of endogenuous opiate system activation and study the roles of opiate polypeptides in the pathogenesis of immunoinjury in virul myocarditis. Methods: Balb/c mice with myocarditis induced by 103 TCID50 coxsackie B3 virus were used as experimental model. Plasma β endorphin levels were determined by radioimmunoassay, endorphin receptor on membrane of cardiac myocyte were measured with radioligand binding technique, cardiac pathology and organ mass (heart,lung) were observed at days 3,7,10,15,25 and 35 after infection. Results:Infection with 103 TCID50 coxsackie B3 virus produced a classic pathological picture of myocarditis in 92.8%(65/70) infected mice. Plasma endorphin concentration was significantly increased at days 3 to 25 postinfection compared to the normal controls. The ratios of liver mass and lung mass to body mass were obviously greater at days 10 to 25 after infection in myocarditis animals than in normal animals, the results might reflect more severe congective heart failure caused by myocardial damage in myocarditis animals. Simultaneously, the increase of binding for endorphin on membrane of cardiac myocytes were detected, but the change in the Kd of receptor were not found in myocarditis mice. Conclusion:The endogenous opiate system is activated in congestive heart failure caused by myocardial damage in murine coxsackievirus B3 my ocarditis, which might affect the cardiac pathogenesis because of the neuroimmunoical modulating effects by opiate polypeptides.

Abstract:Objective: To prepare the monoclonal antibodies against ciliary neurotrophic factor(CNTF). Methods: Cell lines of hybrids secreting monoclonal antibodies against CNTF were established by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with CNTF and cloning 3 times; asitic fluid and serum containing abundant monoclonal antibodies against CNTF were obtained by in vivo induction; then identification of the monoclonal antibodies against CNTF and detection of antibody titer were undergone. Results: Five cell lines of hybrids secreting monoclonal antibodies against CNTF were established. Subtypes of these antibodies were all IgG2a. Antibody titers in asitic fluid and serum were 1∶107-1∶108 and 1∶106, respectively. These monoclonal antibodies showed specificity in demonstrating neurons secreting CNTF in CNS, and one kind of them inhibited the enhancing effect of CNTF on the proliferation of neonal SD rats skeletal muscle cells in culture. Conclusion: The monoclonal antibodies against CNTF with high activity and specificity have been established successfully, and will be a useful tool in the studies of CNTF"s structure,function and mechanism

Abstract:Objectve: To study the relationship between the drugs and blood sugar(BS) after brain injury. Methods: The effect of usual drugs which treating brain injury(BI) on BS was observed in rats after head injury.Results: (1)The group of dexamethasonum ＋ 10％ glucose ＋ insulin, and the group of dexamethasonum ＋ 20％ mannitolum ＋ insulin were effective for treating high BS after drug injection. (2)20％ mannitolum was effective for treating high BS after drug injection. (3)Naloxone could markedly decrease BS. Conclusion: The drugs of mannitolum, naloxone and certain pH could decrease blood sugar in brain injury animals

Abstract:Objective: To explore the effect of 21-aminosteroid (U-74389G) on brain edema post traumatic brain injury (TBI) in rats. Methods: Water content of brain after TBI was assayed by means of dry/wet weight methodology, and the content of ion was measured by an atom absorption spectrum analysis apparatus. Results: The turbulence of sodium, potassium, calcium and magnesium after TBI could be alleviated by U-74389G. The increased water content of injured brain tissue could also be alleviated by the administration of U-74389G. Conclusion: The study indicates that U-74389G could effectively improve brain edema after TBI

Abstract:Objective: To investigate the characteristics of the electron encephalogram (EEG) of the sella turcica region tumor (STRT). Methods: Two hundred and 24 patients with STRT, confirmed by surgery, pathology, CT or MRI, were divided into 3 groups: (1) Pituitary adenoma (153 cases). (2) Craniopharyngioma(46 cases). (3) Other tumors in sella turcica region (25 cases). EEG were recorded with conventional method 2-30 days before operation. Results: Over half of the patients (56.7%) had normal or mild abormal EEGs. The rest of the patients had diffuse abnormal EEGs (29.0%), paroxysmal abnormal EEGs (8.5%) or local abnormal EEGs(4.9%). 24 cases displayed intermittent rhythmic delta activities (IRDA, 10.7%). Seventy-seven cases showed an inhibition of physiological waves. IRDA and inhibition of physiological waves frequently appeared in the EEGs of the craniopharyngioma group. The rate of the abnormality of EEG in the patients with large tumor (D>2 cm) was higher than that of the patients with small tumor (D<2 cm). Conclusion: Non -specific changes in EEG are the predominant manifestations in STRT. The main characteristics of the abnormal EEGs in STRT are diffuse or paroxysmal abnormality. Only a few cases have local abnormal EEGs. The extent of abnormality in EEGs correlates to the size of tumor. EEG is helpful in qualitative diagnosis of STRT

Abstract:Objective：To study the relationship between diet nutrition and the prevalence rate of stroke. Methods：By force of stepwise regression analysis，we analyse the data of prevalence of stroke in 1986 and the cross-sectional study data of diet intake from 1984 to 1986 in 143 survey areas in China. Results: There was a negative association between protein, carbohydrate, calcium and the prevalence rate of stroke. By contrary, there was a positive association between other nutrition factors intake and the prevalence rate of the stroke. Conclusion: The dietary factors have some influences on the prevalence of stroke. High protein and high calcium intake are protective factors of stroke, on the contrary, vitamin A may be a risk factor of stroke.

Abstract:Objective:To determine calcitonin gene related peptide (CGRP) levels in cord plasma and amniotic fluid of pregnancy induced hypertension (PIH) patients. Methods: CGRP levels in cord plasma and amniotic fluid of 26 normal pregnant women (NP group) were measured with radioimmunoassay (RIA).Results: Plasma CGRP levels in umbilical artery of PIH patients were significantly higher than that of NP women (P＜0.01),and it increased as the disease became worse, the severe increased markedly when compared with the mild (P＜0.05).CGRP levels in amniotic fluid of PIH patients were significantly higher than that of NP women(P＜0.05).There was a negative correlation between neonatal 1 min Apgar Scores after delivery and CGRP levels in umbilical artery(P＜0.01),as well as that in amniotic fluid(P＜0.05).Conclusion: The changes of CGRP levels in umbilical artery and amniotic fluid are correlated with intrauterine fetal distress

Abstract:Objective: To use analogtodigital conversion technique to mensurate the time interval in skeletal muscle during excitation-contraction coupling. Methods: Two way infrared detectors, analogtodigital conversion card, Rechert Jung KF-80 quick freezing fixation were employed in this test. Results: The time internal of 0.8,5.6,8.4 ms during the skeletal muscle function change was measured, and the morphological images were synchronously obtained. Conclusion: Analogtodigital conversion technique provides the methodology for the function-morphological study

Abstract:Objective: To study the relation between medicine consumption and the injury severity. Methods: The use of medicine and the injury conditions of soldiers in the rear hospital in the Two Mountains Fight were investigated. And the Abbreviated Injury Scale and Injury Severity Score(AIS-ISS) was applied to quantify the condition of the injuries. Defined Daily Dose(DDD) was introduced as the consumption unit. Results: Analyzed by SAS software, the model between ISS(X) and DDDs(Y) was established: Y=16.99lnX-0.53. r2=0.663 7, P＜0.01.It showed that in the particular scope of ISS, the frequency of antibiotics administration rose up with the increase of the injury severity, and it tended to saturate as the severity went worse. Conclusion: For the first time ISS is used to analyze medical supplies consumption, the multi-factors are transformed into mono-factor affecting medical supplies in wartime. It is a good solution to the problem of how to measure injury severity. The general rule of medicine consumption is discovered in rear hospital. The method is easy use.