Abstract:Objective： To establish a capillary electrophoresis method for the separation and determination of chlorobenzyltetrahydroberberine （CBTHB）enantiomers and to investigate their chiral pharmacokinetics in rabbits. Methods： Racemic CBTHB was separated optimally and measured successfully by capillary electrophoresis, based on comparison of chiral recognition capabilities of carboxymethylated cyclodextrins and selection of other electrophoretic conditions.The plasma concentration data were handled with theory of statistical moment and the pharmacokinetic parameters were obtained. Results： CBTHB enantiomers in rabbit plasma samples were separated under optimal chiral conditions. After a single oral dose of 150 mg/kg racemic CBTHB in six rabbits, plasma level of R-enantiomer was always higher than that of S-enantiomer. The ratio of R-to S-enantiomer was (1.43±0.21), resulting in more significant differences between the plasma concentrations of R-enantiomer and those of S-enantiomer（t = 4.14，P＜0.01）. Furthermore, it was found that AUC0？τ and AUC0？∞ of R-enantiomer were significantly greater than those of S-enantiomer. Conclusion：The new chiral capillary electrophoresis method proves to be suitable for studies on CBTHB enantiomers pharmacokinetics.

Abstract:Objective: To investigate the growth and the immunogenicity-related phenotype of B cell-human hepatoma cell fusion cells after 60Co-radiation. Methods: Growth curve and soft agar colonogenicity of fusion tumor vaccine in vitro, and their tumorigenicity in nude mice in vivo for the radiated cell fusion tumor vaccine were observed. The expression of MHC-Ⅰ,MHC-Ⅱ, B7 and hepatoma-antigen(GP75) that were related to immunogenicity of the radiated fusion tumor vaccine cell were analyzed by flow cytometry. Results: 60Co-radiated fusion cells lost proliferative capacity, partly died on d 4 and almost completely died on d 7,while MHC-Ⅰ,MHC-Ⅱ, B7 and GP75 had no apparent changes in immunogenicity-related phenotype in the first 3 d. Conclusion: 60Co-radiated fusion cells lose proliferative capacity, but they have no apparent changes in immunogenicity-related phenotype in the first 3 d.

Abstract:Objective: To assess the protective mechanism of cold cardioplegia in ischemic hearts by observating effects of different protective methods on ischemic myocardial intracellular energy metabolism and ion homeostasis. Methods: Isolated rat hearts were perfused in a Langendorff preparation with a modified Krebs-Henseleit medium with 3.5 mmol/L Tm(DOTP)5-, a sodium shift agent. Experimental groups underwent 60 min ischemia(groupⅠ) either after a 5 min perfusion of hyperkalemic cardioplegia(groupⅡ) or at 15℃(group Ⅲ), or with both interventions(group Ⅳ), followed by 30 min of reperfusion. Interleaved 31P and 23Na NMR spectra were continuously monitored to measure ATP, Pcr,Phi,Nai and Pi during ischemia and reperfusion. Left ventricular function was monitored with an intraventricular balloon simultaneously. Results: The myocardium showed intracellular acidosis and sodium accumulation severely in group Ⅰ, ATP and Pcr of which being not detected after 15 min of ischemia. After 30 min of ischemia, the changes of group Ⅱ were similar to those of group Ⅰ. The changes of group Ⅲ were more slight compared to groupⅠ and Ⅱ during ischemia. ATP and Nai of group Ⅳ were maintained. The left ventricular function was preserved in group Ⅲ and Ⅳ after reperfusion. Conclusion: Hypothermia as the sole intervention was more effective than hyperkalemic on ischemic myocardial metabolism and function. The combination of hypothermia and hyperkalemia could prevent ischemic myocardium from intracellular acidosis and eliminate sodium accumulation effectively, so it plays a greater protective role in ischemic myocardial function.

Abstract:Objective: In a human leukemic megakaryocytic cell line HIMeg, differentially expressed genes regulated by induced differentiation will be cloned, and from which the key genes in the process of induced differentiation will be identified. Methods: The human megakaryocytic leukemia cell line HIMeg was treated with 13-cis retinoic acid for 4 d, then total cellular RNA was extracted and reverse-transcribed to first strand cDNA. The first strand cDNA was subjected to PCR-based mRNA differential display, cloning and sequence analysis. Results: From 5 cDNA fragment candidates obtained, RNA dot blot analysis demonstrated non-regulation in 3 fragments, undetectable signals in one fragment, and altered gene expression in one cDNA fragment which was designated MDI-1 (mRNA downregulated after induced differentiation).The nucleotide sequence data reported here will appear in the GenBank under the accession number AF026526. Conclusion: A cDNA fragment encoded by a unknown gene was cloned. Further studies are required to determine its full length sequence, gene structure and biological function(s).

Abstract:Objective: To explore whether acycloguanosine (ACV) can inhibit the proliferation and induce the erythrodifferentiation of human myeloid leukemia K562 cells. Methods: K562 cells were cultured with ACV for 4 d to detect cellular changes. Results: When K562 cells grew in a medium containing ACV, the cellular growth and division were gradually suppressed, growth fraction decreased and further differentiation towards the cell producing hemoglobins was found. Conclusion: ACV can inhibit proliferation and induce erythrodifferentiation of K562 cells.

Abstract:Objective: To study the expression of integrins in fibroblasts in patients with scleroderma. Methods: Employing technique of indirect immunocolloidal gold and electron microscope, we observed the expression of integrin α5 ,β1 subunits and the supermicro-structures of sclerderma fibroblasts cultured in medium absorbed with fibronectin. Results: The results showed that the expression of integrin α5 ,β1 subunits in fibroblasts from scleroderma was higher than that in normal fibroblasts (P<0.01), and colloidal golds clustered on the surfaces of scleroderma fibroblasts. The cytoskeletal structure: microfilaments were dense and aggregated into bundles. Conclusion: All of these results suggest that the interactions of fibroblasts with fibronectin mediated by integrin α5 ,β1 subunits increase in scleroderma

Abstract:Objective: To seek evidence for perilymph- cerebrospinal fluid barrier (CSF). Methods: A well-controlled comprehensive comparison of kinetics of hydrocortisone (HC) and methylprednisolone (MP) in perilymph of scala tympani (PST), perilymph of scala vestibuli (PSV), scala media endolymph (SME), CSF and plasma was made in guinea pigs. Results: One hour post intratympanic injection, the mean HC concentrations were exactly the same for both the PST and PSV. The concentrations in the two cochlear partions rapidly decreased from 1 h to 2 h post injection. Thereafter it decreased gradually. HC concentration was undetectable in CSF following direct intratympanic application. Likewise, topical application of MP resulted in the rapid appearance of the substance in both the PST and PSV. The mean value of MP concentration in PSV were similar to those in PSV. There was no significant difference in MP concentrations between the PST and PSV although there appeared to be a trend at 1, 2 and 6 h for concentrations to be greater in PSV than in PST. No detectable concentration of MP in CSF was observed after intratympanic administration. Both the HC and MP levels were significantly higher in SME than in the two perilymphatic compartments throughout the experiment. Conclusion: The findings of the experiment indicate that there is no open communication via the cochlear aqueduct that would allow PST to enter CSF. The CSF cannot directly enter the scala tympani via the cochlear aqueduct, displace and ultimately replace PST. Accordingly, the present study confirms the presence of a tight perilymph-CSF barrier.

Abstract:Objective: To research the huoxuehuayu effects of different cultivated populations of safflower, and probe the intraspecific variance of safflower, and provide the basis for evaluation of the crude drug and breeding of good cultivated populations. Methods: Percentage of restraining platelet aggregation, prothrombin time and partial thromboplastic time were measured to compare the huoxuehuayu effects of different populations of safflower. Results: The huoxuehuayu effects of different populations were significantly different(P<0.01). The effects on restraining platelet aggregation from strong to weak were Jimusa′er population, Jianyang population, Xinxiang population and Weishan population (P>0.05 between the last two populations).The effects on prolonging prothrombin time and partial thromboplastic time from long to short were Weishan population, Jianyang population, Xinxiang population and Jimusa′er population(P>0.05 between the last two populations). Conclusion: The huoxuehuayu effects of different cultivated populations of safflower were different because of the different indexes. Jimusa′er population was best on restraining platelet aggregation. Weishan population was best on prolonging prothrombin time and partial thromboplastic time.

Abstract:Objective: To compare the pharmacokinetics and bioavailability between ambroxol hydrochloride oral solution and tablet. Methods: Volunteers were given oral solution or tablet with a single oral dose of 90 mg ambroxol hydrochloride in a randomized crossover study. The plasma concentrations of ambroxol were determined by HPLC method. Three factors analysis of variance and two one-side t test were adopted in the evaluation of the area under plasma drug concentrations-time curve (AUC ). Results: The maximum plasma concentration(cmax) of ambroxol oral solution and tablet were(169.03±23.42) and (163.17 ± 23.41) ng/ml; the time to reach cmax (tmax) were (1.20±0.22) and (1.44±0.23) h; elimination half-life(t1/2β) were(7.07±1.54) and (7.97 ± 1.17) h, AUC were (1 287.13±250.84) and (1 317.85 ± 151.31) h*ng/ml, respectively. Conclusion: The two preparations of ambroxol are of the same bioavailability. The relative bioavailability of oral solution of ambroxol is (97.61±15.65)%.

Abstract:Objective: To explore the role of the obese protein (OP) of the obese gene expressive product in the genesis and development of noninsulin-dependent diabetes mellitus (NIDDM). Methods: Plasma OP levels were measured by radioimmunoassay in 21 cases of normal subjects, 24 adult obese patients and 20 patients with NIDDM. At the same time, the other parameters were measured, including body mass index, plasma cholesterol, triglyceride, fast blood glucose, hemoglobin A1 and the plasma insulin level during glucose tolerance test. Results: The levels of the plasma OP in NIDDM patients (81.0±17.5) pg/ml were very significantly lower than those in normal subjects (194.3±17.7) pg/ml (P<0.01). The levels of the plasma OP in non-obese NIDDM patients were very significantly lower than those in non-obese normal subjects (P<0.01) and the levels of the plasma OP in obese NIDDM patients were very significantly lower than those in obese patients (P<0.01). Conclusion: The plasma OP may be closely related to the genesis of NIDDM. It is one of the important factors resulting in obesity that the plasma levels of OP were lower.

Abstract:Objective: To observe the metabolism of carnitine in patients with chronic renal failure(CRF).Methods:The plasma free carnitine(FC) concentrations were monitored in the undialyzed and dialyzed patients with CRF and normal subjects.Results: (1)Plasma FC concentration in undialyzed patients with CRF was significantly higher than that in normal subjects(P<0.05),and it was closely correlated with blood urea nitrogen(r=0.666 0) and uric acid(r=0.642 0,P<0.05),not correlated with serum creatinine.(2)Predialysis plasma FC concentration in dialyzed patients was much lower than that in normal subjects and undialyzed patients(P<0.01),postdialysis plasma FC concentration was significantly lower than predialysis concentration(P<0.001). There was a 70%decline in plasma FC concentration during one hemodialysis session.Furthermore,plasma FC concentration was very significantly correlated with dialysis time(r=-0.879 3,P<0.001),not correlated with creatinine clearance rate. Conclusion: There is obvious carnitine metabolism disturbance in patients with CRF,especially in hemodialyzed patients whose plasma FC was deficient.So,it is essential for hemodialyzed patients to be timely supplemented with carnitine after hemodialysis.

Abstract:Objective:To investigate the effect of hypothermia on amino acid neurotransmitters in gerbil models of unilateral cerebral ischemia.Methods:Sixteen Mongolian gerbils were randomly subjected to unilateral internal carotid artery occlusion under conditions of either 37℃ or 32℃ for 30 min.Hippocampal concentrations of glutamate ,glycine and GABA were monitered using in vivo microdialysis combined with high performance liquid chromatography.Results:Hypothermia significantly inhibited the release of glutamate and glycine,therefore the high excitotoxic index seen in the normothermic group was strikingly decreased in the hypothermic group.Conclusion:The neuroprotective effect of hypothermia on cerebral ischemia may reside , in part , in its ability to decrease extracellular excitotoxic indexes.

Abstract:Objective: To study the relation between the changes of five elemental contents in serum and schizophrenia. Methods: The dynamic changes of the trace elements Zn, Cu, Fe and ordinary elements Ca,Mg were examined in 65 patients with schizophrenia by atomic absorption spectrophotometer. Results: (1)The contents of Zn,Mg and Zn/Cu in patients with schizophrenia were significantly lower than those in controls, while Cu,Fe, Ca and Ca/Mg were significantly higher than those in controls.(2)The content of Ca in schizophrenic patients with course of disease ＜3 years was significantly higher than those ≥3 years.(3) After 14 and 21 d treatment with chlorpromazine or clozapine, the contents of Zn, Mg and Zn/Cu increased obviously, while Cu, Fe, Ca and Ca/Mg decreased obviously. Conclusion: These findings suggest that the patients with schizophrenia might have an abnormal serum Zn, Cu, Fe, Ca and Mg metabolism.This would play a role in causing and developing schizophrenia.

Abstract:Objective: To establish a rabbit middle cerebral artery (MCA) embolic stroke model, and to study intra-arterial thrombolysis with recombinant staphylokinase (r-Sak) in this model. Methods: Fifteen rabbits were subjected to a model of RMCA embolic stroke by injected autologous clots into the right internal carotid arterial. Thirty minutes after embolization, the rabbits were treated with saline, r-Sak 0.4 mg/kg or urokinase (UK) 4×104 IU/kg intra-arterial. Digital subtraction angiograms were performed every thirty minutes. Results: RMCA were reopened 100%after injection of r-Sak vs 40%reopened after injection of UK (P<0.01). The ischemia and neurological impairment in both groups were limited. UK prolonged thromb in time, kaolin prothrombin time and prothrombin time, decreased fibrinogen whereas r-Sak did not influence these factors. Conclusion: In the rabbit model of MCA embolic stroke, r-Sak is fibrin-specific and is more effective in lysing embolic clots than UK.

Abstract:目的：了解IFNγ对持续肝外性淤胆所致肝纤维化的作用。方法：制作大鼠持续肝外性淤胆模型，用药组于胆管结扎同时每日肌注IFNγ 5×104 U，用药2周；对照组则仅行胆管结扎而不用药。2周后分别测定血清生化指标、肝纤维化指标及肝能量代谢有关指标。结果：IFNγ可改善血清生化指标(P＜0.01或P＜0.05)。在组织学表现上，用药组肝纤维化程度显著低于单纯结扎组(P＜0.01)。用药组大鼠淤胆后肝羟脯氨酸及血清透明质酸水平升高明显受抑(P＜0.01)。肝ATP及CP测定显示淤胆合并用药时两者均保持于较高水平。结论：IFNγ可有效抑制大鼠持续肝外性淤胆所致肝纤维化