Transcriptomic Profiling of Electroacupuncture Regulating the Molecular Network in Hippocampus of Rats with Cerebral Ischemia-Reperfusion Injury.

Objective: To explore the mechanism of electroacupuncture stimulation of the hand-taiyin meridian in regulating the molecular network of rats with cerebral ischemia-reperfusion injury based on transcriptomics. Methods: Male SD rats were randomly divided into sham operation group, model group, and electroacupuncture (EA) group. Middle cerebral artery embolization/reperfusion injury (MCAO/R) was used to establish the model group and EA group. The sham operation group only performed sham operation without modeling and any intervention, and the model group was bound daily. The EA group received electroacupuncture to stimulate the acupoints of hand-taiyin meridian for 14 days. Then, neurological scores, pathomorphological observations, and Tunel staining were performed. Finally, the affected hippocampus of the rat was used for transcriptome sequencing and RT-PCR detection. Results: After electroacupuncture intervention in rats, neurological function scores were improved, and neuronal apoptosis was reduced. The results of transcriptomics showed that a total of 1097 differentially expressed genes were obtained, of which 422 were upregulated and 675 were downregulated. The bioinformatics analysis showed that those differentially expressed genes were related to axon development, neuron projection development, neuron projection morphogenesis, plasma membrane cell projection morphogenesis, cell part morphogenesis, notch signaling pathway, long-term potentiation, MAPK signaling pathway, Hedgehog signaling pathway, and so on. The results of RT-PCR showed that Caspase 9 mRNA increased and BDNF, Grin2a, and PlexinD1 mRNA decreased after electroacupuncture intervention (P < 0.05). Conclusion: Electroacupuncture intervention on hand-taiyin meridian may reduce neurological function scores, inhibit neuron apoptosis, and enhance neuronal repair neuroreparation in MCAO/R rats, which may be related to the regulation of genes such as Caspase 9, BDNF, Grin2a, and PlexinD1.

[Observation of electroacupuncture intervention on cell apoptosis and Bax and Bcl-2 expression in cerebral and myocardial tissues in cerebral ischemia rats based on "heart-brain correlation" theory].

OBJECTIVE: To compare the effect of electroacupuncture (EA) of "Neiguan" (PC6) of the Pericardium Meridian, "Shenmen" (HT7) of the Heart Meridian，"Shuigou" (GV26) of the Governor Vessel and "Zhaohai" (KI6) of the Kidney Meridian on myocardial and cerebral cell apoptosis in cerebral ischemia (CI) rats, so as to explore its mechanism underlying improvement of CI based on the theory of "Heart-brain Correlation". METHODS: Forty-eight SD rats (half male and half female) were randomly divided into normal control, model, PC6, HT7, GV26 and KI6 groups (n＝8 in each one). The CI model was established by occlusion of the middle cerebral artery (MCAO). EA (4 Hz/20 Hz, 1.5 mA) was applied to the right PC6, HT7, GV26 or KI6 respectively for 30 min, once every 12 h for 5 times. The cell apoptosis of the ischemic myocardial and cerebral tissues was detected by TUNEL method, and the expression of cerebral and myocardial Bax and Bcl-2 was determined by using immunohistochemistry. RESULTS: Following modeling, the cell apoptosis percentages and Bax-positive cells of both myocardial and cerebral tissues were significantly increased in the model group in comparison with the control group (P<0.01). After EA intervention, the cerebral apoptotic percentage and cerebral Bax-positive cells in the cerebral tissue of the PC6, HT7 and GV26 groups, and the myocardial apoptosis percentage in the PC6 and HT7 groups, as well as the myocardial Bax-positive cells in the PC6 group were obviously decreased (P<0.05, P<0.01), while the cerebral Bcl-2 positive cells in the PC6, HT7 and GV26 groups, and the myocardial Bcl-2 positive cells in the PC6 and HT7 groups were significantly increased relevant to the model group (P<0.01, P<0.05). No significant changes were found in the KI 6 group in the cell apoptosis index and percentages and Bax- and Bcl-2-positive cells of both myocardium and cerebral cortex tissues compared with the model group (P>0.05)．. CONCLUSION: EA stimulation of PC6 and HT7 can inhibit CI injury induced cell apoptosis of cerebral and myocardial tissues in CI rats, which is possibly associated with its effects in down-regulating Bax expression and up-regulating Bcl-2 expression of both myocardial and cerebral tissues.

[Clinical observation of warm acupuncture combined with yoga posture method in the treatment of periarthritis with frozen period].

OBJECTIVE: To observe the clinical efficacy of warm acupuncture combined with yoga posture method in the treatment of periarthritis with frozen period. METHODS: Ninety patients with periarthritis who met the inclusion criteria were randomly divided into a control group 1, a control group 2 and an observation group, 30 cases in each group. Warm acupuncture was applied in the control group 1 (Jianzhen (SI 9), Jianyu (LI 15), Jianliao (TE 14), etc were selected), yoga posture method was applied in the control group 2, warm acupuncture combined with yoga posture method were given in the observation group, the treatment was given once a day, 10 times as a course with 2 days between courses and continuous for 2 courses. After 2 courses of treatment, the shoulder joint pain score and shoulder function grading were used to evaluate the clinical efficacy, and the clinical efficacy was observed. RESULTS: ①The pain scores of the three groups were significantly lower after treatment (all P<0.01), and scores in the observation group was better than that in the control group 1 and the control group 2 (P<0.05, P<0.01). There was no significant difference between the control group 1 and the control group 2 (P>0.05). ②After treatment, the functional classification of shoulder joints were significantly improved in the three groups (all P<0.01), and the functional classification of shoulder joint in the observation group and the control group 2 were better than that in the control group 1 (P<0.01, P<0.05). There was no significant difference between the observation group and the control group 2 (P>0.05). ③After 2 courses of treatment, the effective rate of the observation group was 86.7% (26/30), which was better than 70.0% (21/30) in the control group 1 and 76.7% (23/30) in the control group 2 (both P<0.05). CONCLUSION: Warm acupuncture combined with yoga posture method can effectively relieve shoulder pain and improve dysfunction. The clinical comprehensive effect is better than simple acupuncture and yoga posture method.

[Effect of Electroacupuncture Stimulation of Acupoints of Heart Meridian on Brain Connectivity in 27 Healthy Volunteer Subjects].

OBJECTIVE: To observe the changes of brain rest-state functional connectivity following electroacupuncture (EA) stimulation of right Xia-Jiquan (HT 1), Shaohai (HT 3), Lingdao (HT 4) and Shenmen (HT 7) of the Heart Meridian and non-acupoints in healthy volunteers so as to provide an imaging basis for clinical treatment of some related disorders. METHODS: A total of 20 healthy volunteers who met the inclusion criteria were randomly divided into HT-acupoint group (n=10 cases) and non-acupoint group (n=10 cases) according to the visiting sequence. EA (2 Hz, 2-5 mA) was applied to the right HT 1-HT 3, and HT 4-HT 7 (two outputs of the EA stimulator being connected to two acupuncture needle handles) and non-acupoints (4 spots on the imaginary mid-line between the HT and the Pericardium Meridian apart from the 4 acupoints HT 1-HT 3, and HT 4-HT 7) for 6 min. All the subjects underwent resting-state (rs) fMRI using an Achieva 1.5 T magnetic resonance image (MRI) scanner before and after EA stimulation. The bilateral posterior cingulate was used as the seed point and the functional connectivity data was analyzed by using DPARSF software and rs-fMRI data analysis toolkit. RESULTS: After EA stimulation of both acupoints and non-acupoints, a similar functional connectivity change was found in the bilateral postcentral gyrus, right inferior parietal lobule, and right inferior frontal gyrus. After EA of acupoints of the HT Meridian, an enhancement of brain functional connectivity was found in the left inferior temporal gyrus and right parahippocampal gyrus, and the newly activated brain regions were the left precentral gyrus, right precuneus, left angular gyrus, left inferior occipital gyrus and the left insula. After EA stimulation of non-acupoints, an increased functional connectivity was found in the left superior temporal gyrus, which was different from EA of acupoints of HT Meridian. CONCLUSIONS: EA stimulation of acupoints of the HT Meridian can enhance the rs functional connectivity in some brain regions as the postcentral gyrus, inferior temporal gyrus, etc. which is related to mediating information of somatosensory, cognitive, affective, memory, cardiovascular activities, etc. in healthy volunteer subjects. The only difference between the two EA groups is an enhanced functional connectivity found in the left superior temporal gyrus after EA of non-acupoints.

[Effect of Electroacupuncture stimulation of acupoints of the Pericardium Meridian on serum NGF and Nogo-A contents and cerebral NGF and Nogo-A expression in cerebral ischemia rats].

OBJECTIVE: To observe the effect of Electroacupuncture (EA) stimulation of "Tianquan"(PC 2), "Quze" (PC 3), "Neiguan" (PC 6), "Daling" (PC 7) of the Pericardium Meridian on cerebral angiogenesis in cerebral ischemia (CI) rats, so as to reveal its mechanisms underlying improvement of stroke. METHODS: A total of 50 SD rats were equally randomized into normal control, sham, model, EA-Pericardium-Meridian acupoints (EA-PCM) and EA-Lung-Meridian acupoint (EA-LUM) groups. The CI model was established by occlusion of the middle cerebral artery. EA (2-4 V, 20 Hz) was applied to PC 2, PC 3, PC 6, PC 7 and "Tianfu"(LU 3), "Chize" (LU 5), "Lieque" (LU 7), "Taiyuan" (LU 9) of the Lung Meridian for 30 min, once at time-points of 0 h, 6 h, 24 h, 48 h and 72 h, respectively after modeling. Serum nerve growth factor (NGF) and Nogo protein-A (Nogo-A) contents were assayed by enzyme linked immunosorbent assay (ELISA), and cerebral NGF and Nogo-A immunoactivity levels in the ischemic cerebral tissue were detected by immunohistochemistry. RESULTS: (1) Compared to the normal control group, serum NGF and Nogo-A contents, and cerebral NGF immunoactivity level in the model group were significantly increased (P < 0.01). Following EA interventions, serum and cerebral NGF levels were further significantly up-regulated in the EA-PCM and EA-LUM groups (P < 0.01), while serum Nogo-A contents were down-regulated in the two EA groups (P < 0.01). The effect of EA-PCM was markedly superior to that of EA-LUM in up-regulating serum and cerebral NGF levels and down-regulating serum No- go-A level (P < 0.01). No significant differences were found between the normal control and sham groups in serum and cerebral NGF and Nogo-A levels (P > 0.05) , and among the 5 groups in cerebral Nogo-A levels (P > 0.05). CONCLUSION: EA stimulation of acupoints of both Pericardium Meridian and Lung Meridian can up-regulate serum NGF, cerebral NGF expression and down-regulate serum Nogo-A in CI rats, and the effect of Pericardium Meridian is markedly superior to that of Lung Meridian, suggesting a possible better nerve repair effect of EA-PCM acupoints on ischemic brain.

[Efficacy observation on chrondromalacia patellae treated with fire needling technique at high stress points].

OBJECTIVE: To compare the clinical efficacy differences between fire needling technique of filiform needle at high stress points and regular acupuncture on chrondromalacia patellae so as to provide the better therapy for the treatment of this disease. METHODS: Sixty cases of chrondromalacia patellae were randomized into a fire needling group (28 cases) and a routine acupuncture group (32 cases). In the fire needling group, 5 to 6 high stress points were localized according to the symptoms, palpation and imaging condition and were stimulated with fire needling technique of filiform needle. The treatment was given once every two days, 5 treatments made one session. In the routine acupuncture group, the regular acupuncture was applied at Dubi (ST 35), Xiguan (LR 7), Yanglingquan (GB 34), Yinlingquan (SP 9), Zusanli (ST 36), etc. The treatment was given once every day, 5 treatments made one session. Lysholm score, VSA score, patella title angle (PTA) and lateral patella angle (LPA) of the affected knees before and after treatment, as well as the clinical efficacy after treatment were observed in the two groups. RESULTS: After treatment, Lysholm score, VSA score, PTA and LPA were all improved apparently in the two groups (all P < 0.01). After the treatments, the improvements in Lysholm score, VSA score, PTA and LPA in the fire needling group were more obvious than those in the routine acupuncture group (all P < 0.05). The effective rate was 92.9% (26/28) in the fire needling group, better than 87.5% (28/32) in the routine acupuncture group (P < 0.01). CONCLUSION: The fire needling technique of filiform needle at the high stress points relieves the clinical symptoms of chrondromalacia patellae and recovers the biodynamical structure of patellae.

[Effect of muscle-tension-balance acupuncture therapy on the motor function and living ability of patients with drop foot and strephenopodia after stroke].

OBJECTIVE: To compare the difference between muscle-tension-balance acupuncture and conventional acupuncture in the impacts on motor function and living ability of patients with drop foot and strephenopodia after stroke. METHODS: Seventy cases were randomly divided into a muscle-tension-balance acupuncture group (group A) and a conventional acupuncture group (group B), 35 cases in each one. In group A, firstly the acupuncture of weakening technique was applied to Sanyinjiao (SP 6) and Taixi (KI 3) on the musculus extensor side; secondly, the strengthening technique was adopted at Jiexi (ST 41), Shenmai (BL 62) and Yanglingquan (GB 34) on musculus flexor side. In group B, the conventional needling technique was applied to Zusanli (ST 36), Sanyinjiao (SP 6), Xuanzhong (GB 39), etc. The treatment was given once every day in either group, 10 treatments made one session and 3 sessions of treatment were required. The score of Fugl-Meyer motor function of the lower limb on the affected side and Barthel index score were assessed before and after treatment in two groups. RESULTS: After treated for 3 sessions, Fugle-Meyer motor function score and Barthel index score of the patients all increased in two groups (all P < 0.01). The results in group A were better than those in group B (both P < 0.05). The improvements were apparent in the 2nd session of treatment in group A (both P < 0.01), which were superior to those in group B (both P < 0.05). CONCLUSION: Either acupuncture therapy can improve the motor function and living ability of patients with drop foot and strephenopodia after stroke. The muscle-tension-balance acupuncture achieves the efficacy quickly and its efficacy is much better.

[Impacts on repeated common cold for the adults with different constitutions treated by acupoint application in the dog days and the three nine-day periods after the winter solstice].

OBJECTIVE: To observe the impacts on repeated common cold for the adults with different constitutions treated by acupoint application in the dog days (the three periods of the hottest days) and the three nine-day periods after the winter solstice (the three periods of the coldest days). METHODS: One hundred and fifty-two cases of repeated common cold were divided into four zones according to the body constitution. Each zone was sub-divided into a group of the dog days + the three nine-day periods of the coldest days (group A), and a simple group of the dog periods (group B). In both groups, Dazhui (GV 14), Feishu (BL 13), Tiantu (CV 22), Danzhong (CV 17), Zhongfu (LU 1) and Shenshu (BL 23) were selected. In group A, the acupoint application was given on the 1st or 2nd day of the first, second and third periods of the hottest days in 2010, as well as the 1st or 2nd day of the first, second and third periods of the coldest days in 2010 separately. In group B, the acupoint application was only given on the 1st or 2nd day of the first, second and third periods of the hottest days in 2010. The follow-up visit was conducted before the acupoint application in the three periods of the coldest days in 2010 and before the acupoint application in the three periods of the hottest days in 2011. Additionally, the frequency of disease attack and the symptom score in sickness were taken as the observation indices for the efficacy assessment in both groups. RESULTS: (1) In both groups, the attack frequency was reduced obviously in half a year after the three periods of the hottest days for the patients of qi deficiency constitution, yang deficiency constitution and qi stagnation constitution and the clinical symptom score were reduced apparently (all P<0.01), which were superior to those for the patients of phlegm damp constitution (P<0.01, P<0.05). For the patients of phlegm damp constitution, only the clinical symptom score was reduced (P<0.01). (2) In group A, the improvements were received in the attack frequency and the clinical symptom score in half a year after the three periods of the coldest days for the patients of those four constitutions as compared with those before treatment (all P<0.01). In group B, the attack frequency and the clinical symptom score were all back to the level as those before treatment. (3) In group A, in half a year after the three periods of the coldest days, the efficacy for reducing the attack frequency and the improvements in the clinical symptoms were better than those in group B (all P<0.01). CONCLUSION: The acupoint application in the dog days and the three nine-day periods after the winter solstice reduces the attack frequency and relieves the clinical symptoms of common cold for the adults of individual constitutions to different extents. The efficacy can be further improved in the treatment with the acupoint application in the three nine-day periods after the winter solstice half a year after the acupoint application in the dog days.

[Observation of the therapeutic effect on COPD of cold phlegm blocking the lung type at stable stage treated with acupoint sticking therapy in different season].

OBJECTIVE: To systematically observe the clinical effect on chronic obstructive pulmonary disease (COPD) at the stable stage, differentiated as cold phlegm blocking the lung type, treated with acupoint sticking therapy during the dog days and the three nine-day periods after the winter solstice so as to propose the latest clinical idea and theoretic evidence for the treatment of COPD. METHODS: One hundred and fifty cases of COPD at stable stage, which were in accordance with the inclusive standard were randomly divided into three groups, named group A (treatment in dog days and the three nine-day periods after the winter solstice), group B (treatment in dog days) and group C (treatment in the three nine-day periods after the winter solstice), 50 cases in each group. The ingredients (Semen Brassicae, Euphoribia Kansui, Asarum, Rhizome Corydalis, Cinnamon, ginger juice) and doses of herbal medicine plaster were same in each group. The herbal plaster was applied to Feishu (BL 13), Shenshu (BL 23), Dazhui (GV 14), Tiantu (CV 22), Danzhong (CV 17) and Zhongfu (LU 1). In group B, the treatment was given once on the 1st day of each dog-day period, totally 3 treatments were included. In group C, the treatment was given once on the 1st day of each nine-day periods after the winter solstice, totally, 3 treatments were involved. In group A, the treatment was given once on the 1st day of each dog-day period and each nine-day periods after the winter solstice separately, totally 6 treatments were required. The therapeutic effect was evaluated in 4 aspects, named comprehensive clinical efficacy, survival quality (the scores for symptoms, activity limitation and influence on daily life), the attach frequency and pulmonary function. RESULTS: The total effective rate was 88.0% (46/50) in group A, which was superior to 76.0% (38/50) in group B and 70.0% (35/50) in group C separately (P < 0.01, P < 0.001). The results of the attack frequency, clinical symptom score and pulmonary function indices after treatment were all improved apparently as compared with those before treatment in each group (all P < 0.01). All the above indices in group A were improved much apparently as compared with the other two groups (P < 0.01, P < 0.001). Except for the level of forced vital capacity (FVC), the results of clinical symptom score and the other pulmonary function indices in group B were all improved significantly as compared with group C (P < 0.05, P < 0.001). CONCLUSION: Acupoint sticking therapy during different season of the year achieves a superior clinical efficacy for the patients with COPD at stable stage. This therapy can reduce the attack frequency and improve the survival quality and pulmonary function for the patients. It is concluded that the efficacy of the treatment in dog days and the three nine-day periods after the winter solstice is superior to simple dog-day treatment and the treatment in the three nine-day periods after the winter solstice, and the efficacy of dog days treatment is better than that in the three nine-day periods after the winter solstice.

[Clinical evaluation on balanced muscular tension needling method for improving disabled function of stroke patients with spastic paralysis].

OBJECTIVE: To observe the effect of balanced muscular tension needling method for improving disabled function of stroke patients with spastic paralysis. METHODS: One hundred and six cases with spastic paralysis were randomly divided into an observation group (54 cases) with balanced muscular tension needling method and a control group (52 cases) with routine acupuncture method. The observation group was treated by acupuncture at the side of extensor and flexor of limbs; while the control group was treated by acupuncture at Jianyu (LI 15), Quchi (LI 11), Waiguan (TE 5) etc. And the change of muscle strength, muscle tonus, muscle spasticity and range of joint motion were evaluated hefore and after treatment. RESULTS: After 30 days of treatment, the total effective rate of 96.3% (52/54) in the observation group was superior to that of 84.6% (44/32) in the control group (P < 0.01). After treatment muscle strength, muscle tonus, muscle spasticity and range of joint motion were improved in the two groups, hut the observation group was superior to the control group (all P < 0.05). CONCLUSION: Balanced muscular tension needling method can significantly improve the muscle strength, muscle tonus, muscle spasticity and the range of joint motion of the stroke patients with spastic paralysis.

[Blood-letting puncture at 12 Jing-well points of the hand combined with fuming and washing with Chinese herb lotion for treatment of edema of the affected limb in patients of hemiplegia after stroke].

OBJECTIVE: To search for a therapy for edema of the affected limb in patients of hemiplegia after stroke. METHODS: Eighty patients were randomly divided into a treatment group and a control group, 40 cases in each group. The treatment group were treated by blood-letting puncture at 12 Jing-well points of the hand combined with fuming and washing with herbal lotion, and the control group were treated by oral administration of diuretic. The therapeutic effect was observed after treatment for 10 days. RESULTS: The total effective rate was 85.0% in the treatment group and 70.0% in the control group with a significant difference between the two groups (P < 0.05). The cumulative score of symptoms of edema, improvement of skin pitting, damp and cool sensation, the function of limbs and the total score in the treatment group were better than the control group (P < 0.05); the cumulative score of TCM syndrome after treatment, improvement of the symptoms of limb flaccid paralysis, shortness of breath, weakness, dark-pale tongue proper and the total cumulative score were better than the control group (P < 0.05). CONCLUSION: Blood-letting puncture at 12 Jing-well points of the hand combined with fuming and washing with Chinese herbal lotion is effective for edema of the affected limb in patients of hemiplegia after stroke.

[Effects of different point association needling methods on IL-1, sICAM-1 and sVCAM-1 in the patient of cerebral infarction].

OBJECTIVE: To observe effects of acupuncture on serum interleukin-1 (IL-1) and soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in the patient of cerebral infarction. METHODS: Eighty-two cases of cerebral infarction were randomly divided into a treatment group treated by balanced muscular tension needling method, and a control group by traditional needling method. The levels of serum IL-1, sICAM-1 and sVCAM-1 were determined with enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of serum IL-1, sICAM-1 and sVCAM-1 in the patient of cerebral infarction were significantly higher than those in the healthy persons (P < 0.01); there were positive correlation between sICAM-1 or sVCAM-1 level and IL-1 level (r = 0.743, P < 0.001; r = 0.862, P < 0.001). The three indexes improved to a certain extent in the two groups, with the treatment group superior to the control group (P < 0.01). CONCLUSION: The improvement of nervous function by the balanced muscular tension needling method in the patient of cerebral infarction is possibly related with decrease of IL-1, sICAM-1 and sVCAM-1 levels.