ABSTRACT
OBJECTIVE@#To select the optimal method for developing experimental animal model of viral facial paralysis by comparing several inoculation methods.@*METHOD@#One hundred and twenty Balb/c mice were divided into 4 groups, with each group having 30 mice. Group A, the posterior auricular branch of right facial nerve were incised and inoculated with 25 microl HSV-1; group B, 25 microl HSV-1 were inoculated into the posterior aspect of the right auricle by cutaneous scarification; group C, 25 microl HSV-1 were injected into subcutaneous tissue of the posterior aspect of the right auricle; group D, 100 microl HSV-1 were inoculated in the way similar to that of group C. The symmetry of mouse face was observed, and the incidence of paralysis and death were analyzed. The temporal bones of paralyzed mice were serially sectioned and stained with hematoxylin and eosin.@*RESULT@#Thirteen (43.33%) mice developed the right facial paralysis and recovered from it 3-7 days later in group A. Six (20%) mice developed the paralysis and recovered from it 2-9 days later in group B. Group C had no signs of facial paralysis and group D had 1 paralyzed animal. Except for 12 mice in group D, there was no death in the other groups. Nerve swelling was observed in right temporal facial nerve of paralyzed mice. Facial nerve to facial canal cross-sectional area ratio (FN/FC) of the right side was much higher than that of the left side.@*CONCLUSION@#Inoculating HSV-1 into the posterior auricular branch of facial nerve can produce an acute and transient facial paralysis in mice. With the advantage of higher morbidity of facial paralysis and lower mortality in comparison to the other methods, it is an optimal method.
Subject(s)
Animals , Female , Mice , Disease Models, Animal , Facial Nerve , Virology , Facial Paralysis , Virology , Herpesvirus 1, Human , Mice, Inbred BALB CABSTRACT
BACKGROUND: Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE: To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes (CD71) to explore the mechanism of TBL on the modulation of cell immunity.DESIGN: A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING: Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS: The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003. Ten clean male SD rats were selected.METHODS: Lymphocyte was separated from rat inguinal lymph node for culture. Concanavalin(ConA) was used for 72-hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT.The expression of T lymphocyte CD71 was tested by flow cytometer after 48-hour stimulation of phorbol 12,13 -dibutyrate(PDB) or ConA.MAIN OUTCOME MEASURES: The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS: Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation. PDB and ConA-activated T lymphocyte CD71 + expressions were significantly higher than that of blank control group(P<0.01) . CD3+ CD71 + expressions [(62.03±1.51) %,(25.28±1.57) %,(20. 29±1.72)%] activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L)were significantly lower than(72.03±1.28)% of BPS-positive control group (P<0. 05). CD3 + CD71 + expressions activated by PDB under 100 mg/Land 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS-)positive control group(P<0.05). Different concentration of overall alkali TBL had significant down-regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose-effect relationship(P<0. 05). The inhibition on ConA-activated CD71 expression was stronger than that of PDB.CONCLUSION: Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.
ABSTRACT
[Objective]To observe the effects of total alkaloids (TA) of Tongbiling (TBL) prescription on proliferation of T lymphocytes and transferring receptor of T lymphocytes (CD71) in rats and to explore the mechanism of TBL in regulating cellular immunity. [Methods]Isolated lymphocytes from mesenteric lymph node were cultured with various concentrations of TBL TA and canavaline A (ConA) for 72 hours; the proliferation of T lymphocytes was observed by MTT method and CD71 expression rate of T lymphocyte activated by phorbol 12,13-dibutyrate (PDB) or ConA for 48 hours was detected by flow cytometry.[Results]Various concentrations of TBL TA inhibited the proliferation of T lymphocytes activated by ConA and decreased CD71 expression rate of T lymphocyte activated by ConA or PDB in a dose-effect manner. Inhibitory effect of TBL TA on CD71 expression rate of T lymphocyte activated by ConA was superior to that activated by PDB. [Conclusion]TBL TA can inhibit the abnormal proliferation of T lymphocytes and its mechanism may be related to the expression of CD71.
ABSTRACT
AIM: To investigate the effect of cycloheximide on the T cells activation by mitogen in vitro with CD69 expression as activation marker for the application of this drug clinically. METHODS:Lymphocytes were isolated from lymphoid nodes of C57BL/6 mouse. The cells were preincubated with cycloheximide(CHX), 5% serum containing CHX respectively for an hour, then further incubated with polyclonal activators (Con A or PDB). Harvesting the cells after whole incubation for 24 h, we estimated the expression rates of CD69 on T cells by flow cytometry following two-color immunofluorescent staining. RESULTS: The expression rates of CD69 on the T cells preincubated with CHX, serum containing CHX after the stimulation in response to Con A or PDB all showed significant difference with the expression rates of control group, respectively ( P