Objective To explore the optimal ratio of Baitong decoction based on efficacy,clarify its spectrum-effect relationship,and identify its potential quality markers.Methods An ulcerative colitis(UC)model in mice was established using dextran sulfate sodium.The efficacy of Baitong decoction with varying drug ratios was assessed by evaluating the apparent score,pathological score and inflammatory factor changes of UC in each group of experimental animals.The fingerprints of Baitong decoction with different ratios were established by high performance liquid chromatography(HPLC),and the relationship between the content of each substance and its efficacy was analyzed by partial least squares regression to determine the potential quality markers of Baitong decoction.Results Baitong decoction was most effective in relieving ulcerative colitis when the mass ratio of Fuzi,Ganjiang and Congbai was 1∶2∶2.The fingerprinting identified 14 common peaks across 7 ratios,with 9 peaks were found to be associated with the remission of ulcerative colitis by partial least squares regression analysis.Conclusion The optimal ratio of Fuzi,Ganjiang and Congbai for treating UC is 1∶2∶2.The spectrum-effect relationship analysis suggested that the quality markers of Baitong decoction may be the substances represented by peak 2(benzoylaconine),3,5,6,8(mesaconine),9(aconitine),10(hypaconitine),13(10-gingerol)and 14.

Objective:To investigate the potential mechanism of electroacupuncture(EA)at bilateral Fengchi(GB20)in treating cerebral ischemia-reperfusion injury and to provide a scientific basis for future experimental research and clinical applications. Methods:Forty male specific-pathogen-free Sprague-Dawley rats were randomly divided into four groups:a normal group,a normal with EA group,a model group,and a model with EA group,with 10 rats in each group.The normal group received no intervention.The normal with EA group received EA at bilateral Fengchi(GB20).The model group underwent middle cerebral artery occlusion(MCAO)using the suture.The model with EA group underwent MCAO and received EA at bilateral Fengchi(GB20).Cerebral blood flow was monitored using a laser Doppler cerebral blood flow meter.Neurologic damage was assessed using the neurologic deficit score,and motor ability was observed using the CatWalk gait system.The expression of glial fibrillary acidic protein(GFAP)and neuronal nuclei(NeuN)protein,the neuron markers,was detected by Western blotting.The protein expression levels of GFAP and NeuN,as well as the number of positive cells in the motor cortex,were detected using immunofluorescence. Results:Compared to the normal group,the cerebral blood flow values in the model group and the model with EA group decreased by more than 50%during the modeling process(P<0.01)and returned to pre-modeling levels after reperfusion(P>0.05).The neurologic deficit score increased(P<0.05),the average motor velocity decreased(P<0.05),GFAP protein expression and the number of positive cells in the motor cortex increased(P<0.05),and the NeuN protein expression and the number of positive cells decreased(P<0.05)in the model group.Compared to the model group,the neurologic deficit score decreased(P<0.05),the average motor velocity accelerated(P<0.05),GFAP and NeuN protein expression and the number of positive cells in the motor cortex increased(P<0.01)in the model with EA group. Conclusion:EA at bilateral Fengchi(GB20)can reduce neuronal loss and increase GFAP and NeuN protein expression in the motor cortex of rats after ischemia-reperfusion,improve the motor function after ischemic stroke,and accelerate the recovery of balance and stability of the affected limbs.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.

Objective: To observe the effects of electroacupuncture (EA) pretreatment on motor function, cerebral blood flow, cerebral infarction volume, and vascular endothelial growth factor (VEGF) level in middle cerebral artery occlusion (MCAO) model rats. Methods: Twenty-four male Sprague-Dawley rats were randomly divided into a normal group, a model group, and an EA group, with eight rats in each group. The middle cerebral artery ischemia-reperfusion model was established by the suture-occluded method in the model group and the EA group, while not in the normal group. The EA group was pretreated with EA at bilateral Fengchi (GB20) before model preparation, once a day for 30 min each time for a total of 7 d. The changes in the CatWalk gait parameters, modified Bederson neurological deficit score, cerebral blood flow, cerebral infarction volume after ischemia, and VEGF level in the brain tissue of rats in each group were observed. Results: Compared with the normal group, the modified Bederson neurological deficit score in the model group and the EA group increased after modeling (P<0.05), and the CatWalk gait parameters (one-leg stance duration, gait cadence, and gait cycle) were all changed (P<0.05). Compared with the model group, the modified Bederson neurological deficit score in the EA group decreased (P<0.05), and the CatWalk gait parameters improved (P<0.05). Immediately after ischemia, the cerebral blood flow in the normal group was greater than that in the model group and the EA group (P<0.05); after reperfusion, the cerebral blood flow in the EA group was greater than that in the model group (P<0.05). Compared with the normal group, the cerebral infarction volume in the model group and the EA group increased (P<0.05). Compared with the model group, the cerebral infarction volume in the EA group decreased (P<0.05). The expression level of VEGF-positive cells in the rat brain tissue in the model group was higher than that in the normal group (P<0.05), and was higher in the EA group than in the model group (P<0.05). Conclusion: EA pretreatment improves the limb motor function in MCAO model rats, alleviates the symptoms of neurological deficits, promotes the recovery of cerebral blood flow, reduces the cerebral infarction area after MCAO modeling, and increases the VEGF expression in the brain tissue.

Background::Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s dementia. Mitochondrial dysfunction is involved in the pathology of PD. Coiled-coil-helix-coiled-coil-helix domain-containing 2 (CHCHD2) was identified as associated with autosomal dominant PD. However, the mechanism of CHCHD2 in PD remains unclear.Methods::Short hairpin RNA (ShRNA)-mediated CHCHD2 knockdown or lentivirus-mediated CHCHD2 overexpression was performed to investigate the impact of CHCHD2 on mitochondrial morphology and function in neuronal tumor cell lines represented with human neuroblastoma (SHSY5Y) and HeLa cells. Blue-native polyacrylamide gel electrophoresis (PAGE) and two-dimensional sodium dodecyl sulfate-PAGE analysis were used to illustrate the role of CHCHD2 in mitochondrial contact site and cristae organizing system (MICOS). Co-immunoprecipitation and immunoblotting were used to address the interaction between CHCHD2 and Mic10. Serotype injection of adeno-associated vector-mediated CHCHD2 and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration were used to examine the influence of CHCHD2 in vivo.Results::We found that the overexpression of CHCHD2 can protect against methyl-4-phenylpyridinium (MPP+)-induced mitochondrial dysfunction and inhibit the loss of dopaminergic neurons in the MPTP-induced mouse model. Furthermore, we identified that CHCHD2 interacted with Mic10, and overexpression of CHCHD2 can protect against MPP +-induced MICOS impairment, while knockdown of CHCHD2 impaired the stability of MICOS. Conclusion::This study indicated that CHCHD2 could interact with Mic10 and maintain the stability of the MICOS complex, which contributes to protecting mitochondrial function in PD.

Objective To preliminarily reveal the neurovascular effect of electroacupuncture and adrenomedullin (ADM) in cerebral ischemia-reperfusion injury.Methods Rat changes after middle cerebral artery ischemia and reperfusion, and the effect of electroacupuncture and ADM on them were investigated using the neurological deficit score, somatosensory evoked potentials and TTC staining technique.Results Electroacupuncture and ADM can significantly improve the neurological deficit score after cerebral ischemia and reperfusion (P<0.05), but there was no significant difference between the two (P>0.05). The P1-N1 and N1-P2 peak values of somatosensory evoked potentials decreased significantly at 30 min after ischemia (P<0.05), recovered somewhat at 60 minutes and decreased significantly again during reperfusion (P<0.05). Electroacupuncture or ADM during reperfusion could significantly improved somatosensory evoked potentials at the end of experiment (P<0.05). Electroacupuncture and ADM could significantly reduce the size of cerebral ischemia-induced infarct (P<0.05), but there was no significant differencebetween the two (P>0.05).Conclusions Electroacupuncture can reduce neurological impairment and improve brain blood supply after cerebral ischemia and reperfusion. That is similar to the neurovascular effect of ADM.

Objective To investigate the clinical efficacy of Yang’s pricking-cupping bloodletting in treating cervical spondylotic radiculopathy and its effect on trapezius tension.Methods Seventy-eight patients with cervical spondylotic radiculopathy were randomly allocated to treatment and control groups, 39 cases each. The treatment group received pricking-cupping bloodletting and the control group, conventional acupuncture plus cupping. The nerve function was scored, the visual analogue scale (VAS) score was recorded and trapezius tension was measured in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups.Results There were statistically significant pre-/post-treatment differences in the nerve function score, the VAS score and trapezius tension in the two groups (P<0.01). There were statistically significant post-treatment differences in the VAS score and trapezius tension between the treatment and control groups (P<0.05). The total efficacy rate was 94.9% in the treatment group and 89.7% in the control group. There was a statistically significant difference in theclinical therapeutic effect between the two groups (P>0.05).Conclusions Yang’s pricking-cupping bloodletting can effectively improve pain symptoms and trapezius tension in patients with cervical spondylotic radiculopathy.

Objective To investigate the clinical therapeutic effect of electroacupuncture on ischemic stroke by recording the therapeutic effect score and performing transcranial Doppler (TCD) detection in electroacupuncture-treated convalescent stroke patients before and after treatment.Methods Sixty convalescent patients with ischemic stroke were randomly allocated to treatment and control groups, 30 cases each. The control group received routine medication and the treatment group, electroacupuncture in addition. In the two groups, pre- and post-treatment neurological defect scores were recorded and the clinical therapeutic effects were observed after four weeks of treatment. Pre- and post-treatment middle cerebral artery (MCA) pulsation indices (RI), resistance indices (RI) and ratios of systolic to end-diastolic (S/D) blood flow velocity detected by TCD were compared between the two groups. Results The total efficacy rate was 80.0% in the treatment group and 50.0% in the control group; there was a statistically significant difference between the two groups (P<0.01). There was a statistically significant pre-/post-treatment difference in the MESSS score in the two groups (P<0.01). There was a statistically significant post-treatment difference in the MESSS score between the treatment and control groups (P<0.05). There were statistically significant pre-/post-treatment differences in TCD indices (PI, RI and S/D) on the affected side in the two groups (P<0.05). There were statistically significant post-treatment differences in TCD indices on the affected side between the treatment and control groups (P<0.05).Conclusion Electro- acupuncture can promote rehabilitation in convalescent stroke patients.

ObjectiveTo investigate the effects of dopamine D1 receptor (D1R) tool drugs and combined acupuncture and medicine on striatal expressions of D1R and dopamine transporters (DAT) in middle cerebral artery occlusion (MCAO)-reperfusion rats.MethodForty-seven male SD rats were randomly grouped, used to make a model and given corresponding interventions. The materials were taken and fixed six hrs later. Striatal D1R and DAT expressions were detectedby an immunohistochemical method in different groups.ResultThe neurological deficit score was significantly higher in the model group than in the blank group. Electroacupuncture treatment decreased the score significantly (P<0.05). D1R expression was significantly down-regulated in the antagonist, electroacupuncture and electroacupuncture plus antagonist groups compared with the model group (P<0.05). There was no statistically significant difference in D1R expression between the model group and the antagonist or electroacupuncture plus antagonist group (P>0.05). DAT expression was significantly down-regulated in the other groups compared with the model group (P<0.05). There was no statistically significant difference in D1R expression between all groups except the model group (P>0.05).ConclusionCerebral ischemia-reperfusion can result in high D1R and DAT expressions in rat striatum on the ischemic side. Electroacupuncture, D1R antagonists and a combination of the two can significantly down-regulate D1R expression and have a protective effect on the brain. The effects of electroacupuncture and D1R antagonists can not be added to each other. D1R signaling pathway may be one of ways by which electroacupuncture produces a protective effecton the brain.

As a component of traditional Chinese medical therapies, the therapeutic effects of acupuncture for some nervous system diseases have been proven by a large number of clinical and experimental studies. But, the electrophysiological techniques of the commonly used EEG and evoked potentials are still not sufficient to reveal the functional mechanism of acupuncture therapy. The recording technique of microelectrode array (MEA), a kind of electrophysiological technique originated from the overseas biological electrical signal monitoring technique, can be used to record multiple electrical signals of the nervous cells in vivo or in vitro, and maintain the accuracy and stability of the recorded information at the same time, which greatly enriches the means of electrophysiological study. This technique has been already applied in the basic study and clinical treatment abroad, but it is very seldom used in the study of acupuncture field. In order to guide the application of MEA in the research field of acupuncture science, a general survey about the application of MEA technique in vivo was done, and the present situation and prospects of the application of the technique in acupuncture science was briefly analyzed.