Electroacupuncture preconditioning protects against focal cerebral ischemia/reperfusion injury via suppression of dynamin-related protein 1.

Electroacupuncture preconditioning at acupoint Baihui (GV20) can reduce focal cerebral ischemia/reperfusion injury. However, the precise protective mechanism remains unknown. Mitochondrial fission mediated by dynamin-related protein 1 (Drp1) can trigger neuronal apoptosis following cerebral ischemia/reperfusion injury. Herein, we examined the hypothesis that electroacupuncture pretreatment can regulate Drp1, and thus inhibit mitochondrial fission to provide cerebral protection. Rat models of focal cerebral ischemia/reperfusion injury were established by middle cerebral artery occlusion at 24 hours after 5 consecutive days of preconditioning with electroacupuncture at GV20 (depth 2 mm, intensity 1 mA, frequency 2/15 Hz, for 30 minutes, once a day). Neurological function was assessed using the Longa neurological deficit score. Pathological changes in the ischemic penumbra on the injury side were assessed by hematoxylin-eosin staining. Cellular apoptosis in the ischemic penumbra on the injury side was assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling staining. Mitochondrial ultrastructure in the ischemic penumbra on the injury side was assessed by transmission electron microscopy. Drp1 and cytochrome c expression in the ischemic penumbra on the injury side were assessed by western blot assay. Results showed that electroacupuncture preconditioning decreased expression of total and mitochondrial Drp1, decreased expression of total and cytosolic cytochrome c, maintained mitochondrial morphology and reduced the proportion of apoptotic cells in the ischemic penumbra on the injury side, with associated improvements in neurological function. These data suggest that electroacupuncture preconditioning-induced neuronal protection involves inhibition of the expression and translocation of Drp1.

miRNA-186 inhibits prostate cancer cell proliferation and tumor growth by targeting YY1 and CDK6.

microRNAs (miRNAs) are known to be important in tumor initiation and progression. Recent studies have demonstrated that miR-186 is critical in several types of cancer, including human non-small cell lung cancer, bladder cancer and pancreatic ductal adenocarcinoma. However, the functions of miR-186 in prostate cancer (PCa) are still unclear. In the present study, downregulation of miR-186 in PCa cells was detected when compared with the normal prostate cell line. When miR-186 overexpressed in PCa cells, cell proliferation in vitro was evidently inhibited as shown using cell counting kit-8 assays and cell-cycle analysis, and tumor growth in vivo was decreased as shown by tumor growth assays in nude mice. Furthermore, through bioinformatics prediction and biochemical analyses, Yin Yang 1 (YY1) and cyclin-dependent kinase 6 (CDK6) have been proven to act as direct targets of miR-186. These results indicate that miR-186 is a negative regulator in PCa by inhibiting PCa cell proliferation via targeting YY1 and CDK6.

Electroacupuncture preconditioning attenuates ischemic brain injury by activation of the adenosine monophosphate-activated protein kinase signaling pathway.

Electroacupuncture has therapeutic effects on ischemic brain injury, but its mechanism is still poorly understood. In this study, mice were stimulated by electroacupuncture at the Baihui (GV20) acupoint for 30 minutes at 1 mA and 2/15 Hz for 5 consecutive days. A cerebral ischemia model was established by ligating the bilateral common carotid artery for 15 minutes. At 72 hours after injury, neuronal injury in the mouse hippocampus had lessened, and the number of terminal deoxynucleotide transferase-mediated dUTP nick-end labeling-positive cells reduced after electroacupuncture treatment. Moreover, expression of adenosine monophosphate-activated protein kinase α (AMPKα) and phosphorylated AMPKα was up-regulated. Intraperitoneal injection of the AMPK antagonist, compound C, suppressed this phenomenon. Our findings suggest that electroacupuncture preconditioning alleviates ischemic brain injury via AMPK activation.

[Effect of electroacupuncture pretreatment on expression of glucose-regulated protein 78 in hippocampus of rats with cerebral ischemia/reperfusion].

OBJECTIVE: To explore the effects and action mechanism of electroacupuncture (EA) pretreatment on rats with transient cerebral ischemia/reperfusion. METHODS: A total of 144 healthy SD male rats were randomly divided into a sham operation group (group S), an ischemia/reperfusion group (group I/R) and an EA pretreatment group (group EA), 48 rats in each one. The model of cerebral ischemia/reperfusion was established by using 4-vessel occlusion method in the group I/R; after 5 min of cerebral ischemia, the reperfusion was performed. The group EA was treated with EA at "Dazhui" (GV 14) and "Baihui" (GV 20) 5 days before model establishment, 30 min per time, once a day. In group S, bilateral foramen alares were exposed without burning on the vertebral arteries, and bilateral common carotid arteries were unfolded and not occluded. The rats in the group I/R and group EA were sacrificed 6 h, 12 h, 24 h and 48 h after reperfusion and those in the group S were sacrificed at corresponding time to collect hippocampus example. The Western-blot method was used to measure the expression of glucose-regulated protein 78 (GRP 78), and HE staining method was used to count the number of surviving neurons, and TUNEL method was used to measure the number of apoptotic neurons. RESULTS: Compare with the group S, the number of surviving neurons in hippocampus was reduced at each reperfusion time point and the number of apoptotic neurons was increased (all P<0.05) in the group I/R and the group EA; the expression of GRP 78 at each reperfusion time point in group I/R and group EA was increased (P<0.05). Compared with the group I/R, the number of surviving neurons in hippocampus was increased at each reperfusion time point and the number of apoptotic neurons was reduced in the group EA (P<0.05); the expression of GRP 78 at each reperfusion time point was further increased (P<0.05). CONCLUSION: The electroacupuncture pretreatment has obvious cerebral protection on rats with ischemia/reperfusion, which is related with further increasing the expression of GRP 78 in ischemia area, leading to relieved endoplasmic reticulum stress.

[Effect of electroacupuncture pretreatment on apoptotic neurons and expression of GRP 78 and GADD 153 in the hippocampus in rats with global cerebral ischemia/reperfusion injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment on the number of survival neurons and the expression of glucose regulated protein 78 (GRP 78) and growth arrest and DNA damage-inducible gene 153 (GADD 153) in the hippocampus in rats with global cerebral ischemia /reperfusion injury (CI/R), so as to study its underlying mechanism in neuroprotective action. METHODS: SD male rats were randomly divided into 3 groups (n =48 each):sham operation,Cl/R model and EA pretreatment group. Global CI/R model was induced by 4-vessel occlusion (bilateral vertebral artery cauterization and bilateral carotid artery ligation for 5 min, followed by reperfusion). Before modeling, EA pretreatment of "Baihui" (GV 20) and "Dazhui" (GV 14, 2 Hz/15 Hz, 1 mA) was given to rats of the EA pretreatment group for 30 min, once daily for 5 days. At 6, 12, 24 and 48 h after CI/R, the hippocampus tissues of rats in different subgroups were separately sampled to be stained with H. E. method for detecting the number of the survived neurons, stained with TUNEL method for assaying the apoptotic neurons in the CA 1 region, and processed with Western blot (WB) for assaying the expression of GRP 78 and GADD 153 proteins. RESULTS: Compared with the sham group, the number of hippocampal survival neurons was significantly decreased at the time-points of 12 h, 24 h and 48 h after CI/R in the model group (P<0. 05) and was apparently increased by EA pretreatment at 24 h and 48 h (P<0. 05). The number of the apoptotic neurons in the hippocampal CA 1 region at the time-points of 6 h, 12 h, 24 h and 48 h after CI/R was significantly bigger in the model group than in the sham group (P<0. 05), and was obviously decreased at 12 h, 24 h, 48 h after CI/R in the EA pretreatment group (P<0.05). WB detection showed that the expression levels of hippocampal GRP 78 and GADD 153 proteins at the four time-points after CI/R were significantly higher in the model group than in the sham group (P<0. 05). Compared with the model group, hippocampal GRP 78 protein expression levels at the 4 time-points were further markedly up-regulated (P<0.05), while GADD 153 protein expression levels at the 4 time-points were significantly suppressed in the EA pretreatment group (P<0.05). CONCLUSION: EA pretreatment can effectively suppress the number of hippocampal apoptotic neurons and increase survival rate of neurons in CI/R rats, which may be closely associated with its effects in up-regulating the expression of GRP 78 protein and down-regulating the expression of GADD 153 protein in the hippocampus.

[Influence of auricular point sticking on incidence of nausea and vomiting and analgesia effect after gynecological laparoscopy].

OBJECTIVE: To observe the influence of auricular point sticking on incidence of nausea and vomiting and analgesia effect after gynecological laparoscopy, and provide evidence for clinical application of auricular point sticking. METHODS: One hundred and twenty cases of selective gynecological laparoscopy under general anesthesia were randomly divided into an auricular point sticking group and a placebo group, 60 cases in each group. In the auricular point sticking group, the auricular point sticking with vaccaria seeds was applied at Shenmen (TF 4), Wei (CO 4) and Jiaogan (AH 6a) before the operation and 1, 5, 9, 23 h after the operation, which were pressed 5 min each point each time. The two ears were proceeded at the same time. In the placebo group, the same point selection, sticking paste was used as the auricular point sticking group, but no sticking or pressing with vaccaria seeds was adopted. The incidence of nausea and vomiting, the usage rate of tropisetron and morphine within 24 hours of the operation, as well as the score of visual analogue scale (VAS) and other adverse reactions at 2, 6, 10, 24 h after the operation were observed respectively. RESULTS: Compared with the placebo group, the incidence of nausea and vomiting [31.7% (19/60), 16.7% (10/60) vs 58.3% (35/60), 35.0% (21/60)], the usage rate of tropisetron [21.7% (13/60) vs 48.3% (29/60)] and morphine [18.3% (11/60) vs 38.3% (23/60)], the VAS scores at all different time points in the auricular point sticking group were all decreased (all P < 0.05), and no adverse reaction was observed. CONCLUSION: The auricular point sticking could significantly decrease the incidence of nausea and vomiting in patients of gynecological laparoscopy and has positive analgesic effect.

[Effects of transcutaneous electrical stimulation of auricular Shenmen point on postoperative nausea and vomiting and patient-controlled epidural analgesia in cesarean section].

OBJECTIVE: To investigate the effects of transcutaneous electrical stimulation of auricular Shenmen point on postoperative nausea and vomiting and patient-controlled epidural analgesia in cesarean section. METHODS: After IRB approval and informed consent, one hundred and eighty singleton primiparas undergoing elective cesarean section, in Qingdao Municipal Hospital, and Qingdao Hiser Medical Center, from November 2011 to March 2012, were randomly assigned to three groups: transcutaneous electrical stimulation of auricular Shenmen point group (group A, n = 60), transcutaneous electrical stimulation of auricular Eye point group (group B, n = 60) and control group (group C, n = 60). Women of group A received transcutaneous electrical stimulation of auricular Shenmen point (frequency 1.5 HZ) at the time of preoperation, 4, 10 and 22 hours of postoperation for 30 minutes. The strength was controlled by themselves. Women of group B received stimulation of auricular Eye point as group A. Women of group C received pressurization and connected line were the same with group A, but without electrical stimulation. The following indexes was observed: the incidence of postoperative nausea and vomiting (PONV) for 48 hours; the rate of metoclopramide; the visual analogue scale (VAS) score of rest pain, uterine contration pain and dynamic pain at the time of postoperation for 6, 12, 24 and 48 hours (T(1)-T(4)); the total number and effective compressions number of patient-controlled epidural analgesia(PCEA);the dose of analgesia mixture; the anal exhaust time; the volume of postoperative bleeding for 6 hours of postoperation and the other side effects. RESULTS: Compared with group B and group C, the incidence of PONV, the rate of metoclopramide, the VAS score at the time T(1)-T(4), the total number and effective compressions number of PCEA, the ratio of the total number with effective compressions number and the dose of analgesia mixture were decreased in group A (P < 0.05), but no difference compared group B with group C (P > 0.05). The anal exhaust time and the volume of postoperative bleeding for 6 hours of postoperation were no difference in the three groups (P > 0.05). No other side effects were observed. CONCLUSION: Transcutaneous electrical stimulation of auricular Shenmen point can reduce the incidence of PONV and improves analgesia effect of PCEA in postoperation of cesarean section.

[Efficacy and safety of patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in cesarean section].

OBJECTIVE: To study the efficacy and safety of patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in cesarean section. METHODS: A randomized controlled clinical trail was conducted on 180 singleton primiparas (SAS > 30) undergoing selective cesarean section. They were randomly assigned to three groups, i. e., the patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) group (Group A, 60 cases), the patient-controlled sedation with transcutaneous electrical stimulation of auricular eye point group (Group B, 60 cases), and the control group (Group C, 60 cases). Patients in Group A received patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in the operating room. The strength was controlled by patients themselves. The stimulation lasted for 30 min before the epidural puncture till ending the surgery. Patients in Group B received stimulation of auricular eye point. Patients in Group C received pressurization with the same connected line as Group A, but without electric stimulation. The following indices were observed: (1) the bispectral index (BIS), heart rate (HR), mean arterial pressure (MAP), Ramsay sedation score when the women entered the operating room (T0), 30 min after stimulation (T1), at the time after removing the fetus (T2), and by the end of surgery (T3); (2) the concentrations of plasma angiotensin II (AngII) and cortisone (Cor) at the aforesaid time points; (3) the use rates of oxytocin, atropine, and ephedrine; the hemorrhage amount, and the neonatal Apgar score. RESULTS: Compared with Group A, the BIS, the plasma concentrations of AngII and Cor increased at T1, T2, and T3 (P < 0.05), and the Ramsay sedation score decreased (P < 0.05). The HR and MAP increased at T1 (P < 0.05) in Group B and Group C. Compared with T0, the BIS, HR, MAP, and Ramsay sedation score, the plasma concentrations of AnglI and Cor were lowered in Group A at T1 (P < 0.05). There was no statistical difference in the use rates of oxytocin, atropine, and ephedrine; the hemorrhage amount, and the neonatal Apgar score (P > 0.05). CONCLUSIONS: Patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in cesarean section had obvious sedative effects. It had no adverse effects on puerperal or neonates.

[Effects of acupuncture pre-conditioning on apoptosis in hippocampal neurons following ischemia-reperfusion injury in aged rats].

OBJECTIVE: To investigate the effects of acupuncture pre-conditioning on apoptosis in hippocampal neurons following ischemia-reperfusion injury in aged rats. METHODS: A total of 120 senile male Wistar rats aged 19 - 21 months (corresponding to 60-year-old human being) weighting 550 - 710 g were randomly divided into 4 groups (n = 30 each). Cerebral ischemic group: 4-vessel-block was conducted for 4 minutes to establish cerebral ischemic models; Acupuncture pre-conditioning group: electroacupuncture was applied at acupoint Baihui (GV20) with a frequency of 15 Hz and 2 mA for 30 minutes once daily for 5 days. Then the rats received 4-vessel-block for 4 minutes; Sham-operation group: 4 vessels were exposed; Sham-acupuncture group: only electroacupuncture for 5 days without operation. The rats were sacrificed at the end of predetermined duration of reperfusion 12 h, 1, 2, 3 and 7 d respectively. The brains were immediately harvested and hippocampal CA1 region was isolated for (1) light and electron microscopic examinations of hippocampal neurons; (2) detection of apoptotic neurons (TUNEL); (3) determination of caspase-3 protein expression with SABC (streptavidin-biotin-peroxidase complex) immuno-histochemical technique. RESULTS: There were apoptotic neurons in all groups. The numbers of apoptotic neurons and positive neurons of caspase-3 significantly increased in the acupuncture pre-conditioning and cerebral ischemic groups versus the sham-acupuncture and sham-operation groups (P < 0.01). And the numbers of apoptotic neurons and positive neurons of caspase-3 significantly decreased in the acupuncture pre-conditioning group versus the cerebral ischemic group (P < 0.01). CONCLUSION: Acupuncture pre-conditioning can decrease the neuronal apoptosis after ischemia-reperfusion injury through a lowered expression of caspase-3 protein in senile rats.

[Therapeutic effect of compound danshen solution on hemorrhagic shock combined with coagulopathy in rats].

To investigate the effects of complex danshen solution and heparin on the changes of blood coagulation factors in rats with hemorrhagic shock, and to explore the therapy of coagulopathy by compound danshen solution, the rat model of hemorrhagic shock was set up, 40 SD rats were randomized into four groups: sham operation, shock, compound danshen solution and heparin groups, each group was composed of 10 SD rats. Plasma SFMC, TM, ATIII, D-D, t-PA, PAI levels and APTT were detected, incidences of bleeding complications between heparin and danshen group were compared. The results showed that plasma SFMC, D-D levels in shock group were higher but ATIII level in shock group was lower than that in sham operation group, compound danshen solution group and heparin group (P < 0.001), TM levels obviously increased in shock group and heparin group (P < 0.001). There was no significant difference between compound danshen solution and sham-operation groups. Plasma t-PA, D-D levels obviously increased after shock for 2 hours, PAI level reached the peak after shock for 4 hours, but t-PA decreased. After shock for 6 hours, plasma PAI descended, t-PA continually drop in, but PAI and D-D remained in higher levels. Plasma D-D level in heparin group was lower than that in shock group, t-PA level was higher than that in shock group, but there was no significant difference between in heparin and shock groups. Plasma t-PA, PAI and D-D levels in compound danshen solution group were lower than that in shock group. APTT of danshen group was lower than that of shock group and heparin group. Bleeding incidences was 30% in heparin group and 0% in danshen group, respectively. It is concluded that compound danshen solution may used to treat hypercoagulation and hyperfibrinolysis. In comparsion with heparin, danshen posses-ses advantages of safety with less bleeding complication and needs not tight monitor.