Abstract:Objective: Apoptosis of human mast cell line (HMC-1) could be i nduced by tripterine, one of the pure ingredients of tripterygium wilfordii hook . The paper is to explore the cell changes in morphology of apoptotic cells. Methods: On or after exposure to tripterine, cells were observed with microscopy and electromicroscopy. Apoptosis was detected by flowcytometry and T UNEL. Results: Sunflower-like cell (SLC), cell surrounded by formed or forming apoptotic bodies with chromatin condensed or not, were seen in the apoptotic process, which was negtive to trypan blue dye. SLC could occur in 20 minutes after incubated with tripterine and survive for several hours. The f ormation of SLC was at a time- and dose- dependent manner. The results of apop tosis assayed by SLC count were significantly correlated with that mearsured by TUN EL or PI/FCM in first 8 h. The SLC could easily be determined by microscopy in t he condition of culture. The results of SLC counted by inter- or intra-examine r were consistent. Conclusion: The results suggest that SLC is a characteristic morphologic change of apoptotic cells, and can be a good indicat or for HMC-1 cell apoptosis.

Abstract:Objective : To study a new method for treatment of spastic bladd er after spinal cord injury. Methods: By utilizing the characteris tics of sacral plexus root and its physiological anatomy, a new way called diffe rential sacral nerve anterior rhizotomy was designed to treat spastic bladder af ter spinal cord injury. Results: The spasticity of bladder was reduced and the storage and evacuation function of bladder was improved significantly. The bladder function became normal in the follow-up investigation after 6 month s. Conclusion: Differential sacral nerve anterior rhizotormy is an effective method to treat spastic bladder after spinal cord injury

Abstract:Objective: To study the characteristic of primary cells of human renal cell carcinoma(RCC) and expression of the oncogenes. Methods: An improved isolation method was employed to isolate primary RCC cells from 7 fresh human RCC specimens. The expression of some oncogenes in the cells was examined by means of chromosomal pattern analysis and immunohistochemistry. Results:It was found that the primary cells obtained by using this met hod was more and purer than that of the routine methods and bcl-2, c-erbB-2 , c-met, EGFR had low to medium level expression, but p53 had no expression. Conclusion: The study provides a basis for the ex vivo gene th erapy for RCC.

Abstract:Objective: To clone the rat augmentor of liver regeneration (AL R) cDNA and analyse its structure. Methods: The rat ALR cDNA was o btained by using RT-PCR method with total RNA extracted from the regenerating hepatic tissue,then was subcloned into the pGEM-T vector.The sequence of rat A LR cDNA was proved to be correct by sequencing.The rat ALR cDNA and its deduced amino acids sequence were both compared with Genebank database to analyse homol ogy. Results and Conclusion: There isn′t any mutation in cloned r at ALR cDNA.The rat ALR cDNA has higher homology with yeast ERV1 cDNA ,and their homology of DNA and protein is 54.99%and 47.01%respectively. We also find tha t a portion of the rat ALR protein has identity with the binding motif in zinc f inger proteins.

Abstract:Objective: To determine the HGV infection rate and status of sup er-infection with other hepatitis virus in hemodialysis patients. Metho ds: Fifty seven serum samples were collected from hemodialysis patients. T wo sets of primers were designed and synthesized according to the conserved regi on at 5′ non-coding region of HGV RNA sequence. HGV RNA fragment was amplified by reverse transcript-nested PCR, and IgG antibody to HGV was tested by EIA. M eanwhile, HBV DNA and HCV IgG antibody were tested in all samples. Results : Among 57 serum samples detected, 4 samples were positive for HGV RNA(7.0%) , and 2 were positive for HGV IgG antibody (3.5%).Among samples positive for HGV RNA, 2 were positive for HBV DNA, and 2 were positive for HCV IgG antibody, and one of which was positive for both HBV and HCV markers. Those samples positive for HGV RNA were negative for HGV IgG antibody and vice versa. Concl usion: The infection rate of HGV in hemodialysis patients may be higher th an i n healthy blood donors and healthy population. Patients with HGV infection can b e super-infected with HBV and HCV.

Abstract:Objective: To investigate the nongenomic effects of steroids on the glycine uptake in synaptosomes and to further elaborate on the physiological functio n of steroids. Methods: In synaptosomes of cerebellum or spinal co rd loaded with labeled glycine, the uptakes of glycine without and with steroids were measured by liquid scintillation technique. Results: Cortico sterone and progesterone stimulated the glycine uptake in synaptosomes of cerebe llum, but estradiol and deoxycorticosterone could not, which demonstrated that the effects of steroids were specific. Corticosterone, progesterone, estradiol a nd deoxycorticosterone showed no effects on the glycine uptake in spinal cord sy naptosomes. Conclusion: The mechanisms of the rapid stimulating ef fects of corticosterone and progesterone on the glycine uptake in synaptosomes o f cerebellum are nongenomic.

Abstract:Objective: To investigate the effect of transforming growth fact or β1 gene transfer on the oncogene expression. Methods: Positi ve and reverse pL(TGFβ1 )SN were used to transfect PA317 and Ψ2 packaging cel l line by lipofectamine. After G418 selection, the positive cloning and serum of retroviral were obtained. Meanwhile, retroviral serum and positive, reverse TGF β1 gene were used for transfecting hepatocellular carcinoma cell line. Pos itive cell slide for immunohistochemical staining was prepared to observe change s in oncogene expression. Results: PL(TGFβ1 )SN was digested b y restriction enzyme. Positive and reverse fragments were formed. 290 bp Neo gene band was discovered by nested primer RT-PCR in all the samples. Conc lusion: Positive TGFβ1 gene reduces the c-myc, c-met and ras gene exp ression suggesting that TGFβ1 could act as a negative growth regulator for hu man hepatocellular carcinoma cells in vitro.

Abstract:Objective: To observe inhibitory effect of intercellular adhesio n molecule-1(ICAM-1) antisense DNA on ICAM-1 expression in endothelial cells. Methods: ICAM-1 antisense DNA vector was constructed and transfe cted into endothelial cells by lipofectamine reagents. ICAM-1 expression was me asured by flow cytometry in endothelial cells and transfected cells with or with out TNFα. Results: Agarose gel electrophoresis showed 1.4 kb ICAM -1 DNA was inserted into pcDNA3 expression vector in reverse direction. ICAM-1 expression in normal cells with the stimulation of TNFα was significantly high er than without the stimulation(P<0.01). There was no significant increase in ICAM-1 expression in transfected cells with the stimulation of TNFα(P> 0.05). ICAM-1 expression in transfected cells was lower than that in normal cel ls(P<0.05). Conclusion: ICAM-1 expression in endothelial cell s can be inhibited by ICAM-1 antisense DNA in vitro.

Abstract:Objective: To study the main distrib ution and the influence of S2？S4 nerves on urethral sphincter, detrusor u rinae and penis. To supply the theoretical guidance for the treatment of spas tic bladder after spinal cord injury with differential sacral rhizotomy. Methods: Twelve dogs were respectively given electrical stimulation to S2？S4 nerve roots. The contraction of urethral sphincter and detrusor uri nae, the change of bladder pressure and penis erection were examined and recorded. The peak value of evoked motor unit potentials of the corresponding muscles w ere compared. Results: Electrical stimulation to S2 nerve root induc ed peak value of evoked motor unit potential of detrusor urinae and change of bladder pressure which amounted to 60%of that under normal condition. Electrica l stimulation to S3 nerve root induced peak value of evoked motor unit potential of musculus sphincter urethrae which amounted to 57%of that under no rmal condition. Electrical stimulation to S4 nerve root induced improved penis er ection. Conclusion: S2？S4 nerves are the main innervating nerves to detrusor urinae, urethral sphincter and penis respectively. This finding helps to further rehablitate and recover the function of the lower urinary tract a fter spinal cord injury and avoid unnessesary functional damage in the course of treatment.

Abstract:Objective: To investigate the relationship between constituents and pharmacological effects of Scrophularia ningpoensis (XuanSheng). Methods: The isolation and identification of compounds were done mainly by no n-polar macroreticular resin, sephadex LH-20, ODS, silica gel column chromatography and FABMS, NMR etc, spectra. Results: The inhibition of the LTB 4 f ormation in the rat abdominal neutral leukocytes by drugs isolated from S. nin gpoensis were observed. The iridoid glycosides XS-6, XS-7 showed weak action , but the phenylpropanoid glycosides XS-8, XS-10 showed potent action at the c oncentration of 0.5 mmol/L. In the inhibition of the rabbit platelet aggregation, iridoid and phenylpropanoid glycosides were active in inhibiting the rabbit threshold ADP-induced and AA-induced but not 20 nmol/L PAF-induced pl atelet aggregation at the concentration of 0.5 mmol/L in vitro. Conclusion: The phenylpropanoid and iridoid glycosides may be associated with anti-inflammation and anti-platelet aggregation effects of S. ningpoensis.

Abstract:Objective: To study the effects of fisetin on proliferation and collagen synthesis of immortalized rat hepatic stellate cell line：HSC-T6 cells . Methods: Cell proliferation was measured by crystal violet stain ing assay. Collagen synthesis was determined by 3H-proline incorporation. Results: Fisetin (6.25-50.00 μmol/L)markly inhibited PDGF-stim ulat ed proliferation and TGFβ1-stimulated collagen synthesis of HSC-T6 cells in a concentration-dependent manner. Conclusion: Fisetin could inhi bit actived hepatic lipocyte proliferation and collagen synthesis

Abstract:Objective: To investigate resource of Aralia taibaiensis and study its reproductive methods. Methods: Resource survey and cu ltivation test. Results: Abundant resource of A.taibaiensis was found to grow indigenously in China, and was distributed in mountainous area of Qinling Mountain, Dabashan Mountain and their range in the midwest. Mean while, the natural multiplicative difficulty to its wild population was als o revealed,which may be artificially settled by either seed reproduction or vege tative propagation. Conclusion: Replacing natural reproduction by seed reproduction and vegetative propagation is an effective method to deal with the shortness of its natural resource.

Abstract:Objective: To evaluate the antipsychotic effects of olanzapine. Methods: The effects of olanzapine on amphetamine induced forced action and 5-hydroxytryptophan (5-HTP) induced head twitches were observed in mice. The experiments of catalepsy induced by olanzapine were carried out in r ats. The results were compared with those of clozapine and haloperidol. Results: Olanzapine (2.4-9.6 mg/kg, ig, ED50 5.88 mg/kg), clozapin e (12-48 mg/kg, ig, ED50 37.16 mg/kg), haloperidol (0.225-0.900 mg/kg , ig, ED50 0.74 mg/kg) could significantly antagonize amphetamine-induced f orced action in mice in dose-dependent manner. Olanzapine (0.3-1.2 mg/kg, ig, ED50 0.54 mg/kg), clozapine (1.5-6.0 mg/kg, ig, ED50 3.10 mg/kg), haloperidol (0.225-0.900 mg/kg, ig, ED50 0.71 mg/kg) coul d significantly antagonize 5-HTP induced head twitches in mice. In rats cataleps y tests, the ig, ED50 of olanzapine, clozapine, haloperidol were 26, 149, 0.54 mg/kg, respectively. Conclusion: Olanzapine possesses dopa mine and 5-HT antagonist activity in vivo, and was a more potent 5-HT anta gonist than dopamine antagonist. Olanzapine has a low property for producing ex trapyramidal symptoms. Olanzapine is expected to find a place as a valuable alt ernative in classical antipsychotics. The combination of olanzapine and haloper idol may be a good recipe for some patients.

Abstract:Objective: To study the effect of fluconazole on ergosterol bios ynthesis in fungi by the method of thin-layer chromatography (TLC). Methods: Saponification and extraction of the nonsaponifiable lipids （NSLs） of fungi were carried out after 24h of incubation with varied concentrations of fluconazole. The components of the NSLs were separated by TLC and the concentr ations of ergosterol and lanosterol were determined. Results: Fluc onazole inhibited ergosterol biosynthesis and caused the concentration of lanost erol increased in a dose-dependent manner in fungi. When fluconazole inhibited ergosterol biosynthesis by 85%, the growth of fungi was also inhibited by about 85%. Conclusion: The effect of antimycotic agents on ergosterol biosynthesis in fungi can be studied by determining the concentrations of ergoster ol and lanosterol by TLC. The method can be used to study the activity and mecha nism of antimycotic agents because of its accuracy and easy operation.

Abstract:Objective: To improve positive rate of thymoma diagnosis. Methods: Antibodies to CAE (citric acid extract of human skeletal muscle ) were evaluated by Western blotting and ELISA respectively in sera of myastheni a gr avis patients (16 cases with and 17 cases without thymoma),normal adults(10 case s) and other disease patients (18 cases). Results: The rates of CA E-Abs to 3.4×104, 5.0×104, 6.0×104 ingredients of CAE-antigen in MGT patients were 93.7%(15/16),75.0%(12/16), 75.0%(12/16) r espect ively. None of other groups was binding to each of these 3 ingredients. The posi tive rates of MGT patients were 16/16,12/16 respectively. The false positive rat e s were 0/17,1/17 respectively. Conclusion: Western blotting assay was more sensitive and specific than ELISA in evaluating CAE-Abs.But the operation of Western blotting assay is more complicated and expensive than ELISA.

Abstract:Objective: To evaluate the effect of extra-panretinal photocoagulation with argon laser for neovascular glaucoma indu ced by central retinal vein occlusion. Methods: Twelve cases (twelve eyes) of neovascular glaucoma were treated by extra-panretinal photocoagulat ion with argon laser. The average number of photocoagulation spots of each eye was 2 050. Results: After photocoagulation, the iris and anterior chamber angle neovascularizations of 8 cases disappeared. The IOPs of 4 cases became normal. Though the IOPs of another 8 cases were still high and anti-glaucoma medicines or other procedures were needed, the IOPS were much lower than that before laser treatment and the ocular pain and headache were disappeared or reduced.Conclusion: The extra-p anretinal photocoagulation appears to be the first choice for neovascular glauco ma induced by central retinal vein occlusion. The earlier and more extensive the photocoagulation is, the better the effect is.

Abstract:Objective: To study the electrophysiologic characteristics and the radiofrequency catheter ablation (RFCA) efficacy of permanent junctional re ciprocating tachycardia(PJRT). Methods: Seven patients (3 men , 4 women) with diagnosis of PJRT were confirmed at electrophysiologic study. Radiofrequency energy was applied at the site of atrioventricular annulus with the earliest retrograde atrial activation during tachycardia or right ventricular pacing. The accessory pathway was ablated to block its conduction using routine RFCA method. Results: All patients had typical attacks of sup raventricular tachycardia with normal heart function, some of them were incessant. During tachycardia long R-P′ intervals, (196±24) ms, were found, and R-P ′/ P′-R ratios were mostly greater than 1 with retrograde Pwaves negative in the inferior leads. Electrophysiologic studies demonstrated all patients with concealed slow conducting accessory pathway for retrograde conduction. At the abl ation sites V-A intervals were (163±22) ms, and decremental conducting properties could be found. Seven accessory pathways were located in right posteroseptal in 4 patients, right midseptal in 1, left posteroseptal in 1 and left posterior in 1. All ablations were successful at the first operation with a mean of (11± 3) min of X-ray time, and without adverse effects. During the follow-up, all patients remained free from tachycardia. Conclusion: PJRT is a special type of atrioventricular reentry tachycardia through concealed slow and decremental conducting accessory pathway. Catheter ablation is an effective and safe curative treatment.

Abstract:Objective: To study the changes of ven tricular myosin light chain (VMLC) and its relation with heart remodeling in rhe umatic heart disease (RHD). Methods: Using Western blotting hybrid izat ion assay, we detected VMLC-1，VMLC-2 in 13 patients with RHD and 7 cases of accidental deathes. Results: As compared with the normal group, the contents of VMLC-1 , VMLC-2 were all decreased significantly in RHD patients, especially the VMLC-2. The ratio of VMLC-1/VMLC-2 in RHD patients varied from 1∶0.95 to 1∶0.61. Conclusion: The values of VMLC-1, VMLC-2 and the ratio of VMLC-1/VMLC-2 are all changed during the development of heart remodeling i n RHD patients, suggesting that these changes may exert a direct effect on the progressive deterioration of heart function.

Abstract:Objective: To study correlation between CR1 genomic density polymorphism of erythrocytes and ability of erythrocytes adhering to tumour cells in variant population(normal people, patient with HBV infection, with hepatoc irrhosis and with hepatocarcinoma).Methods: Genomic determination of the CR1 density polymorphism (HH type, HL type, LL type) of erythrocytes was p erformed using polymerase chain reaction amplification and Hind Ⅲ retriction e nzyme digestion. Ability of erythrocytes adhering to tumour cells using tumour erythrocyte rosetest. Results: In different population,ability of HH type erythrocytes adhering to tumour cells was significantly higher than that of HL type erythrocytes. Ability of HL type erythrocytes adhering to tumour cells was significantly higher than that of LL type erythrocytes. In same CR1 genomic type population, ability of erythrocytes adhering to tumour cells in normal p eople significantly higher than that in patients with HBV infection and patients with hepatocirrhosis and patients with hepatocarcinoma. Conclusion: These results indicate that activity of erythrocytes adhering to tumour cells may be related to CR1 genomic density polymorphism type.

Abstract:Objective: To investigate the rule of allicin induced apoptosis.Methods: Terminal deoxylnucleotide transferase directed X-UTP nick and end labelling (TUNEL) assay, DNA gel electrophoresis and light microscopy were used to observe apoptosis. Results: Trypan Blue staining positive rates of OVCA-3 cell were under 18.5%after being exposed to allicin at t he d osage 0.01-100 μg/ml for 2 h and 10 μg/ml for different time. Allicin induced apoptosis of OVCA-3 cells at 1-100 μg/ml for 2 h. Apoptosis peak was shown at 50 μg/ml. Apoptotic cells accounted for 65.9%of the population. Conclusion: Allicin of appropriate dosage induces apoptosis of OVCA-3 cel l, while has little toxicity on OVCA-3 cell. That is important for clinical app lication of allicin.