[Effects of surgical operation on helper-inducer T-lymphocytes in sino-nasal neoplasms patients].

OBJECTIVE: To explore the effects of surgical operation on helper-inducer T-lymphocytes (Th1/Th2) in sino-nasal neoplasms. METHODS: From January 2004 to December 2011, 80 patients with malignant tumor in nasal cavities or sinuses were enrolled as experimental group and another 80 subjects with deflection of nasal septum as control group. The phorbol-12-myristate-13-acetate (PMA)-stimulated peripheral blood mononuclear cells (PBMC) were obtained from both malignant tumor and normal control tissues. And flow cytometry was used to detect the expression percentages of interleukin-4 (IL-4) in Th2 cell and interferon-gamma (IFN-γ) in Th1 cell at pre and post-operation. RESULTS: The preoperative expression percentage of CD3(+) CD8(-) IL-4(+) cell in malignant tumor patients was higher than that in controls (6.6% ± 1.7% vs 2.8% ± 1.7%, P < 0.05) while the expression percentage of CD3(+) CD8(-) IFN-γ(+) cell was lower (18.7% ± 5.7% vs 59.3% ± 1.5%, P < 0.05). The preoperative expression percentage of CD3(+) CD8(-) IL-4(+) cell was higher than that at postoperation (6.6% ± 1.7 %vs 2.8% ± 1.5%, P < 0.05). And the postoperative expression percentage of CD3(+) CD8(-) IFN-γ(+) cell was higher than that at preoperation (54.0% ± 4.0% vs 18.7% ± 5.7%, P < 0.05). CONCLUSION: Surgery may restore the immune balance in patients with malignant tumors in nasal cavities or sinuses.

[Electroacupuncture at "Baihui"（GV20） and "Yongquan"（KI1） improves learning-memory ability of APP/PS1 double transgenic mice by regulating endosomal-lysosomal system].

OBJECTIVE: To explore the mechanism of electroacupuncture（EA） in improving learning-memory ability in Alzheimer's disease （AD） mice from the perspective of endosomal-lysosomal system. METHODS: Male APP/PS1 transgenic mice were randomly divided into model group and EA group （n=10 in each group） and 10 male C57BL/6 wild mice were taken as the normal group. EA （1 Hz/50 Hz, 1 mA） was applied at bilateral "Yongquan"（KI1） and acupuncture was applied at "Baihui" （GV20） for 15 min. The mice of the model and normal groups were subjected to restriction with the same method as those of the EA group for 15 min. The treatment was conducted once every other day for 6 weeks. The spatial learning-memory ability （shown by escape latency of place navigation test and the time of crossing the target platform and total swimming distance in the target quadrant in 1 min of spatial probe test ） was detected by Morris water maze test. The immunoactivity of senile plaques （SP） in the hippocampus tissue was detected by immunohistochemistry. The ultrastructural characters of hippocampal neurons were observed by transmission electron microscope, and the expression levels of Ras-related protein 5 （Rab5）, Ras-related protein 7 （Rab7） and cathepsin D （CTSD） in the hippocampus were detected by Western blot, separately. RESULTS: Compared with the normal group, the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD were significantly increased （P<0.05, P<0.01）, while the number of crossing the original platform and the total swimming distance in the platform quadrant were considerably reduced （P<0.05） in the model group. In contrast to the model group, the EA group had a marked decrease in the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD （P<0.05, P<0.01）, and a striking increase in the number of crossing the original platform and the swimming distance in the platform quadrant （P<0.05）. Results of transmission electron microscope showed an accumulation of endosome, lysosome, and endolysosomes in the hippocampal neurons in the model group, which was evidently milder in the EA group. CONCLUSION: EA of GV20 and KI1 can improve the learning-memory ability of AD mice, which may be related to its function in reducing hippocampal Aß deposition and down-regulating endosomal-lysosomal system activity.

[Effects of electroacupuncture on the expression of ß-amyloid and autophagy-related proteins in hippocampal cells of mice with Alzheimer's disease].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on the expression of ß-amyloid (Aß) and autophagy-related proteins in hippocampal cells of Alzheimer's disease (AD) model mice, so as to explore its underlying mechanisms. METHODS: Eighteen male APP/PS1 transgenic mice (6 months old) were randomly divided into model and EA groups, with 9 mice in each group. Nine male C57BL/6 wild-type mice of the same age were chosen as the normal group. Mice in the EA group were treated with acupuncture on "Baihui" (GV20) and EA (1 Hz/50 Hz, 1 mA) on bilateral "Yongquan" (KI1), once every other day, 20 min each time for a total of 21 times. After the interventions, the spatial learning and memory ability were observed by Morris water maze test. The autophagy-related pathological changes in hippocampus were observed by transmission electron microscopy. The expressions of microtublue associated protein 1 light chain 3 (LC3) and Aß in hippocampus were observed by immunofluorescence and the expression levels of LC3 and p62 proteins were detected by Western blot. RESULTS: Compared with the normal group, the escape latency was prolonged (P<0.01), the residence time in the original quadrant platform was shor-tened (P<0.05), the positive expressions of LC3 and Aß, the expression levels of LC3Ⅱ and p62 proteins, and the ratio of LC3Ⅱ/LC3Ⅰ proteins in hippocampus were increased (P<0.01, P<0.05) in the model group. Compared with the model group, the escape latency was shortened (P<0.05), the residence time in the original quadrant platform was prolonged (P<0.05), the positive expressions of LC3 and Aß, the expression levels of LC3Ⅱ and p62 proteins, and the ratio of LC3Ⅱ/LC3Ⅰ proteins in hippocampus were decreased (P<0.05) in the EA group. The transmission electron microscopy showed that the structure of neurons was normal in the normal group, a large number of autolysosomes and autophagosomes existed in hip-pocampal nerve cells in the model group, and only a small number of autophagosomes were observed in the EA group. CONCLUSION: EA can reduce the expression levels of autophagy-related proteins LC3 and p62 in APP/PS1 transgenic mice, improve the hip-pocampal autophagy state, reduce intracellular Aß aggregation, and thus improve the learning and memory ability.

[Effects of electroacupuncture at "Baihui" and "Yongquan" on the levels of synaptic plasticity related proteins postsynaptic density-95 and synaptophysin in hippocampus of APP/PS1 mice].

OBJECTIVE: From the perspective of ß-amyloid (Aß) toxicity and synaptic plasticity, the mechanism of electroacupuncture to improve learning and memory ability in the early pathological stages of Alzheimer's disease was explored. METHODS: Twelve male amyloid-protein precursor (APP)/γ-secretase (PS1) double transgenic AD mice were randomly and equally divided into electroacupuncture (EA) group and model group, and other 6 male C57BL/6 mice were used as the normal group. EA (1 Hz/50 Hz, 0.5 mA) was applied to "Baihui" (GV20) and bilateral "Yongquan"(KI1) for 15 min, once every other day for 6 weeks. Immunofluorescence was used to observe the positive expression of Aß in the left hippocampus. Immunohistochemistry was used to observe the positive expression of postsynaptic density-95 (PSD-95) in the left hippocampus. Western blot was used to detect the expression of PSD-95 and synaptophysin (SYN)in the right hippocampus. RESULTS: Immunofluorescence results showed that extracellular Aß was seen in the model group and electroacupuncture group, but no senile plaques were seen. Compared with the normal group, the expression level of Aß in the hippocampus of the model group increased significantly (P<0.01). Compared with the model group, the expression of Aß in the hippocampus of the EA group decreased (P<0.05). Immunohistochemical results showed that compared with the normal group, the PSD-95 positive expression in the model group was decreased(P<0.05). Compared with the model group, the expression of PSD-95 in the EA group was increased (P<0.05). Western blot results showed that compared with the normal group, the expression levels of PSD-95 and SYN in the hippocampus of the model group were decreased (P<0.05, P<0.01). Compared with the model group, the expression levels of PSD-95 and SYN in the EA group were increased (P<0.05，P <0.01). CONCLUSION: EA can reduce the expression of Aß in the hippocampus of APP/PS1 mice and increase the expression of PSD-95 and SYN, which may contribute to its effect in improving the synaptic plasticity.

[Effect of electroacupuncture stimulation of "Baihui" (GV 20) and " Yongquan" (KI 1) on expression of hippocampal amyloid-ß and low density lipoprotein receptor-related protein-1 in APP/PS 1 transgenic mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) treatment on the level of hippocampal amyloid-beta peptide (Aß) and its key transport receptor low density lipoprotein receptor-related protein-1 (LRP 1) in APP/PS 1 transgenic mice so as to explore its mechanism underlying improvement of Alzheimer's disease (AD). METHODS: Twenty-four male APP/PS 1 transgenic mice were equally and randomly divided into model group and EA treatment group, and 12 C 57 BL/6 mice were used as the normal control group. EA (1 Hz/50 Hz, 0.3 mA) was applied to "Baihui" (GV 20) and "Yongquan" (KI 1) for 15 min, once every other day for 6 weeks. The learning-memory ability was detected by using Morris water maze testing, left hippocampal Aß 1-40 and Aß 1-42 contents were assayed by ELISA, and right hippocampal LRP 1 expression was detected using Western blot (WB). RESULTS: Results of Morris water maze test showed no significant differences among the three groups in the escape latency, the times of the platform-site crossovers, the time spent in the target platform quadrant (P>0.05). Compared with the model group, the moderately increased escape latency had a decreasing tendency in the EA treatment group. ELISA assaying showed that hippocampal Aß 1-42, Aß 1-40, and ratio of Aß 1-42/Aß 1-40 of the model group were significantly higher than those of the normal control group (P<0.01). After EA intervention, the increased Aß 1-42 , Aß 1-40, and ratio of Aß 1-42/Aß 1-40 were remarkably down-regulated in the EA treatment group (P<0.01). WB detection displayed that the right hippocampal LRP 1 expression level of the model group was markedly lower than that of the normal control group (P<0.05). After EA treatment, LRP 1 expression level was moderately up-regulated but without significant difference between the model and EA treatment groups (P>0.05). CONCLUSION: EA intervention can lower the level of hippocampal Aß in APP/PS 1 transgenic mice, but its effects on Aß transport receptor LRP 1 expression and learning-memory ability need being confirmed further.

[Effect of electroacupuncture intervention on autophagy pathway in APP 695 V 717 I transgenic mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) therapy on the intraneuronal Abeta1-42 and dysfunction of autophagy pathway, so as to reveal its mechanism underlying improvement of Alzheimer's disease (AD). METHODS: APP 695 V 717 I transgenic female mice were randomly divided into model group (n = 6) and EA group (n = 6); and C 57 BL/6 mice were used as the control group (n = 6). After 3 months' treatment by EA therapy at "Baihui" (GV 20) and "Yongquan" (KI 1) (15 min, once every other day, 2 Hz/100 Hz, 1-2 mA), the expression level of Abeta1-42 of the striate cortex was detected by immunohistochemistry. TUNEL staining was used to detect the degree of apoptosis of the striate cortex, and ultrastructural changes of autophagosome in the cortex were observed using electron microscope. RESULTS: In comparison with the control group, Abeta1-42 expression level and the apoptotic neurons in the striate cortex were significantly up-regulated in the model group (P < 0.01). Following EA intervention for 3 months, the Abeta1-42 expression level and the number of apoptotic neurons were significantly decreased in the EA group (P < 0.01, P < 0.05). Accordingly, transgenic induced dark degenerated neurons exhibiting irregular body deformation, analosis, and abundant secondary lysosomes and autophagosomes were reduced in the EA group. CONCLUSION: EA intervention can effectively down-regulate Abeta1-42 expression and number of the apoptotic neurons in the striate cortex in APP transgenic model mice, which may contribute to its effect in improving pathological changes of ultrastructure of neurons.

[Effect of electroacupuncture on learning-memory ability, and Abeta and LRP1 immunoactivity in hippocampal sulcus microvessels in APP transgenic mice].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) of "Baihui" (GV 20) and "Yongquan" (KI 1) on the expression of amyloid beta protein (Abeta) and low density lipoprotein (LDL) receptor-related protein 1 (LRP 1) in the hippocampal sulcus microvessels in amyloid precursor protein (APP) 695 V717 I transgenic mice, so as to study its mechanism underlying relief of Alzheimer Disease (AD). METHODS: Twelve APP 695 V717 I transgenic mice were randomly divided into model group and EA group. Six C 57 BL/6 mice were used as the control group. EA (2 Hz/100 Hz, 3-5 mA) was applied to "Baihui" (GV 20) and "Yongquan" (KI 1) for 15 min, once every other day for three months. The learning-memory ability of mice was detected by using Lashley III water maze system. The expression level of Abeta(1-42), and LRP 1 in the hippocampal sulcus microvessels were measured by immunohistochemistry. RESULTS: Water maze test showed that the swimming duration from the start to the goal box (terminal) in the Lashley III water maze was significantly longer in the model group than in the control group (P < 0.05), suggesting a markedly lower learning-memory capacity of APP 695 V717 I transgenic mice. Compared with the model group, the swimming duration in the EA group was decreased considerably (P < 0.05). The integrated optical density (IOD) value of hippocampal Abeta(1-42) immunoreaction (IR) positive products in the model group was significantly higher than that in the control group (P < 0.01) and the IOD value of hippocampal LRP 1 IR-positive products in the model group was apparently lower than that in the control group (P < 0.01). In comparison with the model group, the IOD value of Abeta(1-42) IR-positive products in the EA group was obviously lower than that in the model group (P < 0.05), while that of LRP 1 IR-positive products in the EA group was significantly higher than that in the model group (P < 0.05), suggesting down-regulation of hippocampal Abeta(1-42) expression and up-regulation of LRP 1 expression after EA, and reduction of deposition of Abeta in the cerebral microvessels after EA. CONCLUSION: EA can improve the learning-memory capacity of APP transgenic mice, which is closely related to its effects in up-regulating hippocampal LRP 1 expression and down-regulating hippocampal Abeta(1-42) expression.

[Effect of electroacupuncture on the behavior and hippocampal ultrastructure in APP 695 V 717 I transgenic mice].

OBJECTIVE: To investigate the ultrastructural basis underlying electroacupuncture (EA) induced improvement of Alzheimer disease (AD) in transgenic mice. METHODS: Twelve APP 695 V 717 I transgenic mice were randomly divided into model group and EA group; and other 6 negative transgenic mice (C 57 BL/6 J) were made up of normal control group. After 3 months treatment by EA (15 min per other day, 2 Hz/100 Hz, 3-4 mA) applied to "Baihui" (GV 20) and "Yongquan" (KI 1), the learning and memory ability of mice was measured by Lashley III water maze test, and the ultrastructural changes of hippocampal CA 1 region was observed by electronic microscopy. RESULTS: The swimming escape latency and the number of navigating errors (dead-end forward swimming) in model group were significantly longer and more than those in normal control group (P < 0.05); and those in EA group were considerably shorter and fewer than those in model group (P < 0.05), suggesting an improvement of learning-memory ability after EA. Comparison of the ultrastructure of the neurons in the hippocampal CA 1 region showed swelling of the mitochondria, broken or disappearance of the mitochondrial cristae, degeneration of the synapses, breakage and vague outline of the basement membrane of the blood capillaries in mice of model group; and basically distinct outline of the mitochondrial cristae and microvessels, and more synaptic vesicles in EA group. CONCLUSION: EA may effectively improve the learning-memory capacity of the APP transgenic AD mice and alleviate the pathological changes of neurons of the hippocampal CA 1 region, which may be one of the mechanisms underlying the improvement of AD by EA.

[Effect of electroacupuncture on the behavior and the expression of amyloid beta-protein, amyloid precursor protein and ChAT in APP 695 V 717 I transgenic mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of amyloid precursor protein (APP) and amyloid beta-protein (Abeta) in hippocampal CA 1 area and cerebral cortex in APP 695 V717 I transgenic mice, so as to investigate its underlying mechanism in improving Alzheimer's disease (AD). METHODS: APP 695 V 717 I transgenic mice were randomly divided into model and EA groups, with 6 cases in each. The other 6 negatively-transgenic mice (C 57 BL/6 J) were used as the normal control group. EA (2 Hz/100 Hz, 3 mA) was applied to "Baihui" (GV 20) and "Yongquan" (KI 1) for 15 min, once every other day for 3 months. The learning-memory ability of mice was measured by Y-type maze test and the expression levels of APP, Abeta, and choline acetyltransferase (ChAT) in the cerebral cotex and hippocampal CA 1 area were detected by immunohistochemistry. RESULTS: In comparison with control group, the times of training for gaining correct reaction and the expression levels of Abeta and APP in both cerebral cortex and hippocampal CA 1 area in model group increased significantly (P < 0.05, P < 0.01), while the expression level of ChAT of cerebral cortex in model group decreased considerably (P < 0.05). Compared to model group, +he times of training for gaining correct reaction and the expression levels of APP in both cerebral cortex and hippocampal CA 1 area, and Abeta in CA 1 area of EA group lowered remarkably (P < 0.05), while those of ChAT in cerebral cortex and hippocampal CA 1 area of EA group increased evidently (P < 0.05, P < 0.01). CONCLUSION: Electroacupuncture can improve the learning-memory capacity of the APP transgenic mice, which may be related to its effects in increasing the production of acetylcholine and lowering the levels of APP and Abeta in the brain.