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Objective To observe the consistency of the effect directions of electroacupuncture(EA)at the"Ciliao"(BL32)and EA on the auricular concha(AC)in regulating intragastric pressure(IGP)and heart rate variability(HRV),and to observe whether the combined EA of"Ciliao"(BL32)and AC has a synergistic effect.Methods SD male rats were divided into EA-AC group,EA-BL32 group and EA-"AC+BL32"group,with 8 rats in each group.And the IGP and HRV were measured before and during EA(30 Hz,2 mA,pluse width 0.2 ms±30%,continuous wave)for 2 min in each group.Results ①Changes in IGP compared with before EA:the IGP in EA-AC group significantly increased especially during the 30th to 60th(P<0.05)and 60th to 90th second of EA(P<0.05);The IGP in EA-BL32 group significantly increased especially during the 60th to 90th second of EA(P<0.05);The IGP in EA-"AC+BL32"group significantly increased especially during the 60th to 90th(P<0.01)and 90th to 120th second of EA(P<0.05).The change rate of IGP of EA-"AC+BL32"group was significantly higher than that of the EA-AC group especially during the 60th to 90th(P<0.01)and 90th to 120th second of EA(P<0.01),and EA-BL32 group especially during the 60th to 90th(P<0.05)and 90th to 120th second of EA(P<0.05).②Changes in HRV compared with before EA:the high frequency(HF)significantly increased(P<0.01,P<0.05,P<0.01,P<0.05),low frequency(LF)significantly increased(P<0.01,P<0.01,P<0.05,P<0.05),and very low frequency(VLF)significantly increased(P<0.05,P<0.05,P<0.05,P<0.05)throughout the EA in EA-AC group;The HF significantly increased(P<0.05,P<0.05,P<0.05,P<0.05),LF significantly increased(P<0.05,P<0.05,P<0.05,P<0.05),and VLF significantly increased(P<0.05,P<0.05,P<0.05,P<0.05)throughout the EA in EA-BL32 group;The HF significantly increased(P<0.05,P<0.05,P<0.05,P<0.05),LF significantly increased(P<0.001,P<0.05,P<0.01,P<0.01),and VLF significantly increased(P<0.05,P<0.01,P<0.05,P<0.05)throughout the EA in EA-"AC+BL32"group.The LF/HF in EA-AC group significantly decreased especially during the 0 to 30th(P<0.01),30th to 60th(P<0.05)and 60th to 90th second of EA(P<0.05);The LF/HF in EA-BL32 group significantly decreased especially during the 0 to 30th second of EA(P<0.05);The LF/HF in EA-"AC+BL32"group significantly decreased especially during the 30th to 60th(P<0.05),60th to 90th(P<0.01)and 90th to 120th second of EA(P<0.01).The change rate of LF/HF of EA-"AC+BL32"group was significantly lower than that of the EA-AC group especially during the 90th to 120th second of EA(P<0.05),and EA-BL32 group especially during the 60th to 90th(P<0.05)and 90th to 120th second of EA(P<0.05).Conclusion The effect direction of EA at"Ciliao"(BL32)and EA on the AC in regulating the vagus nerve is basically the same,and the combined EA of"Ciliao"(BL32)and AC can produce a synergistic effect.
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Objective To evaluate the effect of intracerebroventricular injection of 7-nitroindazole (7-NI) on the depression-like behaviors in normal rats.Methods According to body weights,48 SD rats were randomly divided into normal group,model group,sham-operation group and 7-NI groups at different concentrations (n=8).The model group was treated with chronic and unpredictable mild stress.The 7-NI groups received intracerebroventricular injection with 7-NI solutions at different concentrations,once every 3 days for 3 times in total.The sham-operation group was injected with DMSO of the same volume.The rat behaviors were then subjected to the open field test (OFT).The hippocampal nNOS protein levels were detected by Western blot.Results Compared with the normal group((132.47±31.72) m),the total movement distances of model group ((15.04±8.61) m) and 200 nmol/0.5 μl surgery group((18.18± 11.82) m) decreased significantly (P＜ 0.05).Compared with the model group,such distances of sham-operation group ((107.33±20.35)m) and 7-NI groups(50 nmol/0.5 μl:(138.40±56.85)m,(86.97±36.20)m);100 nmol/0.5 μl:(86.97±36.20)m) increased significantly (P＜ 0.05).The normal group entered the central area significantly more times(2.25±2.05) than model group (0.25±0.46)and 200nmol/0.5μl 7-NI group (0.25± 0.46) did (P＜0.05),and the number of times entering the central area of the model group (0.25±0.46)was significantly lower than that of the sham-operation group (1.00 ± 1.07,P＜ 0.05) and 50 nmol/0.5 μl group (0.75 ± 1.16).Compared with the normal group ((46.53 ±41.16) s),the durations of stay in the central area of model group ((1.27 ± 1.92) s) and 200 nmol/0.5 μl 7-NI group ((1.53 ± 2.90) s) were shortened significantly (P＜0.05).Compared with the model group,the durations of stay in the central area of 100 nmol/0.5μl group ((36.54±67.80) s) was lengthened significantly (P＜ 0.05).Western blotting showed that the hippocampal nNOS protein levels of model group (0.43±0.11) and 200 nmol/0.5μl 7-NI group(0.56±0.08) significantly exceeded that of the normal group (0.04±0.02,P＜0.05).The levels of nNOS in sham-operation group (0.04 ±0.02) and 50 nmol/0.5 μl 7-NI group (0.22± 0.08),which were significantly lower than that of the model group (0.43 ± 0.11,P＜ 0.05),were similar to that of the normal group (0.04 ± 0.02,P＞ 0.05).Conclusion Intracerebroventricular injection 200 nmol/0.5 μl 7-NI solution results in depression-like behaviors and increased the expression of nNOS protein reflexively in rats.
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To analyze the structural specificity of Shenque (CV 8) in terms of vascular biology from the three aspects of structure, tissue and molecular anatomies. On structural anatomy Shenque (CV 8) possessed defined vascular structure and was the only acupoint directly affected vascular intima. The basis of the specific therapeutic effects was owing to its relation with vessel and abundant microcirculation. Endothelial cells and microvascular endothelial cells were the tissue basis of the starting of therapeutic specificity. Molecular anatomy involved in the functions of transient receptor potential vanilloid family (TRPV) pathway and neuronal peptide secretion of endothelial cells in the structural function of the point. The "restoringfor collapse" effect of salt-partition moxibustion at Shenque (CV 8) was analyzed based on the local biological specificity of blood vessels. The mechanisms were concluded as effectively targeting the dysfunction of microvascular endothelial cells and acquiring the maximum quantity of moxibustion by repeated warm-heat stimulation. The vascular biological structural features of Shenque (CV 8) may contribute to direct therapeutic effects on endothelial cells by the point.
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Objective To observe the temperature-time curve of Shenque(CV8) salt moxibustion and explore the temperature starting mechanism of Shenque moxibustion and itsclinical significance.Method In the subjects treated with Shenque salt moxibustion, periumbilical and umbilical bottom temperatures were measured simultaneously and recorded using a two-channel temperature logger, and the temperature-time curve was drawn and analyzed to preliminarily explore its temperature starting mechanism according to the experimental results.Result Periumbilical temperaturemeasurement showed that the temperature that every cone of moxa produced reached more than 43℃and the maximum temperature reached 44.9℃. The time that 43℃or more lasted was (4.50±2.36) min. The time that 40℃or more lasted was (8.03±2.63) min. Umbilicalbottom temperature measurement showed that the temperature reached more than 43℃at the seventh cone of moxa on average and the maximum temperature reached 44.7℃. The mean value of the maximum temperatures at the seventh to tenth cones of moxa was (43.44±1.05)℃. The time that 43℃or more lasted was (5.53±2.13) min. The time that 40℃or more lasted was(18.44±10.39) min.Conclusion Shenque salt moxibustion has the property of repeated warm stimulations, which can avoid the platform period brought about by stable warm stimulation and promote the maximization of moxibustion intensity to effectively increase clinical moxibustion quantity finally.
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BACKGROUND:Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cel transplantation, but the mechanism is stil unclear. OBJECTIVE:To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions. METHODS:(1) Bone marrow mesenchymal stem cel s were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem cel s was tested by x-gal staining. (2) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cel scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions for 6 hours. RESULTS AND CONCLUSION:(1) Transfection efficiency of bone marrow mesenchymal stem cel s was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradual y increased with hypoxia time (P<0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P<0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P<0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cel s was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P<0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cel s can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cel s under hypoxia and serum-free conditions.
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BACKGROUND:Adrenomedulin gene transfection can strength the anti-apoptotic ability of bone marrow mesenchymal stem cels under ischemia and hypoxia, but its mechanism is not yet clear. OBJECTIVE:To investigate the effect of adrenomedulin on the expression of apoptosis-related proteins, Bax, Bcl-2 and Caspase-3, in bone marrow mesenchymal stem cels under hypoxia and ischemia. METHODS:Bone marrow mesenchymal stem cels of Sprague-Dawley rats were isolated, cultured and purified, and then cultured in serum-free medium under hypoxic condition for 0, 3, 6, 9, 12 hours. Then, western blot assay was employed to detect the expression of Bax, Bcl-2 and Caspase-3 so as to determine the optimal hypoxia time that was determined at 6 hours of hypoxia. Depending on whether adrenomedulin pretreatment was done, the cels were divided into control group (with no adrenomedulin pretreatment before hypoxia and ischemia) and adrenomedulin groups with different concentrations (1, 10, 100 μg/L). Afterwards, the expression of Bax, Bcl-2 and Caspase-3 was detected by using western blot assay. RESULTS AND CONCLUSION:(1) After cultured in serum-free medium under hypoxia for 0, 3, 6, 9, 12 hours, the expression of Bax, Bcl-2 and Caspase-3 in bone marrow mesenchymal stem cels were increased (P < 0.05);at 6 hours of hypoxia, the Bax/Bcl-2 ratio and Caspase-3 expression reached the minimum value (P < 0.05). (2) At 6 hours of hypoxia, the expression of Bax and Caspase-3 protein as wel as Bax/Bcl-2 ratio became the lowest in the 100 μg/L group compared with the 1 and 10 μg/L groups, but the expression of Bcl-2 protein reached the peak (P < 0.05). These findings indicate that adrenomedulin can reduce the expression of Bax/Bcl-2 ratio and Caspase-3 protein in bone marrow mesenchymal stem cels cultured in serum-free medium under hypoxic conditions, which is in a dose-dependent manner.
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@# Firstly effect of opposing needling was underlying acupoints of uninjured side facilitate injured side, similar as the nature of association reaction. That hinted association reaction may be the mechanism of opposing needling. Then opposing needling about its clinical application indication, therapeutic effect, stimulus intensity and strategy was discussed and interpreted on the basis of apprehension association reaction conception, essential characteristic and its role in hemiplegia rehabilitation.