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To observe the efficacy of electroacupuncture (EA) plus passive stretch exercise in the treatment of disused atrophy of gastrocnemius and soleus muscles in mice.Fifty C57BL/6 mice were randomly and equally divided into 5 groups: blank control, model, passive stretch exercise (exercise), EA and EA+exercise groups. The muscular atrophy model was established by fixing the gastrocnemius and soleus muscles with plaster immobilization (by putting the right leg into a plastic vial and then twining the vial with medical plaster bandage from the ankle upwards to the thigh and groin to maintain the knee-joint flexion and ankle joint plantar flexion for 7 days). EA (2 Hz/100 Hz, 1 mA)was applied to bilateral "Zusanli"(ST36) for 10 min, once a day for 4 weeks. For mice with the passive exercise, the plastic vial was removed first, followed by pulling out the hindleg to seize the toes to stretch them until the right hindleg is fully extended, then, pushed the leg towards the body. The procedures were repeated once again and again for 10 min. The exercise was conducted once daily, for 4 weeks. The cross-sectional area of fast and slow muscle fibers of the soleus and gastrocnemius was measured under electronic microscope after ATPase histochemical stain and the expression of slow skeletal muscle troponin (TNNI1) and fast skeletal muscle troponin (TNNI2) in the soleus and gastrocnemius was detected by Western blot.Compared with the blank control group, the cross-sectional areas of the fast and slow muscle fibers of the soleus and gastrocnemius muscles were significantly decreased in the model group (EA combined with passive stretch exercise can promote the recovery of the soleus and gastrocnemius muscles in disused muscle atrophy mice, which may be related to its effect in up-regulating the expression of TNNI1 and TNNI2 proteins.

This study is aim to investigate the effect of electro-acupuncture at PC6 (Neiguan-point) on the gene and protein expressions of classical inward-rectifier potassium channels (Kir) in myocardial ischemia (MI) rats induced by isoproterenol (ISO). With ten for each one, 50 rats were divided into 5 groups which were control group, MI group, PC6 group, LU7 (Lieque-point) group and non-acupoint group. The control group was injected normal saline solution (85mg/kg), the other groups were injected ISO (85mg/kg). All the rats were injected once daily for two days and recorded electrocardiograms (ECGs) after every injection. Electro-acupuncture (EA) was operated at PC6, LU7 and non-acupoint respectively in the rats of PC6 group, LU7 group and non-acupoint group after twice injections. EA was performed to these three groups with disperse-dense wave (4-20Hz), pulse amplitude of 14V, 20mins a day remaining 7 days. The gene and protein expressions of Kir2.1, Kir2.2 and Kir2.3 were analyzed by Western Immunoblotting Technology (Western Blot) and Real-time Fluorescence Quantitative Polymerase Chain Reaction (RT-PCR). But it is regrettable that we did not detect meaningful gene and protein expressions Kir2.3, and the expressions of Kir2.1 and Kir2.2 in MI induced groups were lower [The gene and protein decreased 39.4 ± 27.3% and 38.7 ± 17.1% respectively.] than control group (PPP < 0.05). The results show that PC6 has a better positive effect than LU7 on MI rats, and the mechanism is probably that EA at PC6 can significantly increase the gene and protein expressions of Kir2.1 and Kir2.2.

To observe the effect of electroacupuncture (EA) on the hormones derived from the hypothalamus-pituitary-ovary (HPO) axis, so as to explore the neuroendocrine mechanism induced by EA on rats with perimenopausal depression disorder.Sixty female sprague-dawley rats were randomly divided into blank control group, model group, sham-operation (sham) group, clomipramine group, and electroacupuncture (EA) group, with 12 rats in each group. Perimenopausal depression model was established by bilateral ovariectomy combined with chronic unpredictable stimulation.The EA group received continuous treatment at "Baihui" (GV 20), "Shenshu" (BL 23) and "Sanyinjiao" (SP 6) once a day for 28 days. Estrous cycle and sucrose preference test were monitored, and serum estradiol (ECompared to the blank control group, sugar water consumpution rates decreased in the model group and sham group (Electroacupuncture can relieve the symptoms of rat with perimenopausal depression by regulating the hormone secretion in HPO axis.

Objective To examine the effect of electroacupuncture (EA) at bilateral Neiguan (PC 6) on voltage-gated Na+ currents (INa) and channels (Nav) in ischemic ventricular myocytes. Methods EA serum was prepared from six male adult Sprague-Dawley rats that had received EA at bilateral Neiguan (PC 6). Eighteen ventricular myocytes were prepared from six SD rats using an enzy-molysis approach. Myocardial ischemia was mimicked by perfusion of ischemic solution. Whole-cell patch-clamping was used to record three currents evoked from isolated cells. The first current was the control, and recorded in absence of ischemic solution current. The second was the ischemic current, and recorded after perfusion of ischemic solution for 5 min, while the EA current was last, and recorded after perfusion of EA serum for 5 min. Nav kinetic curves were fitted using related formulas. Results Compared with those in controls, in the presence of ischemic solution, peak amplitudes of INa significantly increased from – 40 mV to +30 mV, and half-maximal inactivation potentials of Nav increased significantly, while half-maximal activation potentials, slope factors and the recovery time from inactivation to activation of Nav were unchanged. Compared with those in the ischemic solution, in the presence of EA serum, peak ischemic current amplitudes significantly reduced from −40 mV to +40 mV, and half-maximal inactivation potentials were restored, while half-maximal activation potentials, slope factors and the recovery time from inactivation to activation of Nav were unchanged. Conclusion EA at bilateral Neiguan (PC 6) can reduce enhanced INa via restoration of delayed Nav inactivation in ischemic ventricular myocytes.

In this study, various types of industry corrosion addition practices are investigated with regard to the ultimate strength performance of bulk carriers. Five types of corrosion additions, namely p...

This research explored and identified the protein composition of rat kidneys after acupuncture at the Taixi acupoint (KI3). Twelve adult male Wistar rats were randomly divided into a control group (n = 6) and an acupuncture group (n = 6). Rats in the acupuncture group received electroacupuncture on the bilateral KI3 for seven days. The kidneys were perfused with ice-cold saline and all kidney proteins were isolated. After protein sample preparation, two-dimensional gel electrophoresis (2-DE) was performed. The interesting spots were analyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). There were nine protein spots with three-fold up-regulation in the kidney after the acupuncture. NAD-dependent isocitrate dehydrogenase and quinone reductase, the proteins involved in energy metabolism, the reduction of endogenous quinones, chemoprotection, and electrophilic stress, were identified. The data indicated that acupuncture at the KI3 of the kidney meridian of the foot shaoyin was able to increase NAD-dependent isocitrate dehydrogenase and quinone reductase expression in the kidney, and supported the relationship between the kidney and KI3.

Under circumstances of heat stress, heat shock transcription factor 1 (HSF1) plays important roles in heat shock protein expression. In this study, an increasing concentration of dithiothreitol (DTT) was found to either enhance or inhibit the heat-induced trimerization of HSF1, suggesting the involvement of dual redox-dependent HSF1 activation mechanisms. Our in vitro experiments show that the heat-induced bonding between the cysteine C36 and C103 residues of HSF1 forms an intermolecular disulfide covalent bond (SS-I bond) and that it directly causes HSF1 to trimerize and bond to DNA. Gel filtration assays show that HSF1 can form intermolecular hydrophobic interaction-mediated (iHI-m) noncovalent oligomers. However, the lack of a trimerization domain prevents HSF1 activation, which suggests that iHI-m noncovalent trimerization is a precondition of SS-I bond formation. On the other hand, intramolecular SS-II bond (in which the C153, C373, and C378 residues of HSF1 participate) formation inhibits this iHI-m trimerization, thereby preventing SS-I bond formation and DNA binding. Thus, HSF1 activation is regulated positively by intermolecular SS-I bond formation and negatively by intramolecular SS-II bond formation. Importantly, these two SS bonds confer different DTT sensitivities (the SS-II bond is more sensitive). Therefore, a low concentration of DTT cleaves the SS-II bond but not the SS-I bond and thus improves DNA binding of HSF1, whereas a high concentration DTT cuts both SS bonds and inhibits HSF1 activation. We propose that these interesting effects further explain cellular HSF1 trimerization, DNA binding, and transcription when cells are under stress.

To observe the effect of electroacupuncture (EA) stimulation of "Neiguan" (PC 6), etc. on expression levels of myocardial chloride (CL-) channel-related genes and intracellular protein kinase C (PKC) protein in myocardial ischemia (M) rats.Seventy SD rats were randomly divided into control group (n = 10), model group (n = 15) , Neiguan (PC 6) group (n = 15), Lieque (LU 7) group (n = 15) and non-acupoint group (n = 15). The MI model was established by i. p. of isoproterenol (ISO, a sympathomimetic beta adrenergic agonist). Electroacupuncture stimulation was applied to bilateral "Neiguan" (PC 6), "Lieque" (LU 7), or non-acupoint [the mid-point between "Tianshu" (ST 25) and "Shenque" (CV 8)] for 15 min, once a day for 7 days. Quantitative RT-PCR was employed to detect the expression levels of cystic fibrosis transmembrane conductance regulator (CFTR, a CL-channel) mRNA and chloride channel calcium-activated 1 (CLCa 1, a member of the family of calcium-activated chloride channels, CLCa) mRNA in the left cardiac ventricle tissue, and Western blot was used to detect the expression level of myocardial PKC protein of the left ventricle.Compared with the control group, the expression levels of myocardial PKC protein, and CLCa 1 and CFTR genes were significantly increased in the model group (P0.05). In comparison with the model group, the expression levels of myocardial PKC protein, and CFTR mRNA and CLCa 1 mRNA in the Neiguan group, and PKC protein and CLCa 1 mRNA in the Lieque and non-acupoint groups, as well as CFTR mRNA in the Lieque group were notably down-regulated (P0.05). No significant change was found in the expression of CFTR mRNA in the non-acupoint group (P0.05), and no significant differences were found between Neiguan and Lieque groups in the expression levels of PKC protein (P0.05). The effects of "Neiguan" (PC 6) were obviously superior to those of non-acupoint in down-regulating myocardial PKC protein, CLCa 1 mRNA and CFTR mRNA (P0.05).EA stimulation of "Neiguan" (PC 6) can down-regulate the expression of myocardial PKC protein, CFTR and CLCa 1 genes in Ml rats, which may contribute to its effect in protecting rnyocardium from ischemic injury.

Bisphenol A (BPA), generally known as bisphenols, has been identified as a potential estrogenic substance. BPA must be conjugated to carrier protein and BSA was commonly used. 4,4-Bis(4-hydroxyphenyl) valeric acid (BHPVA) has a bisphenolic structure and a long carbon chain with a reactive carboxyl group on the end. In this study, BHPVA-BSA was used to produce polyclonal antibody against bisphenolic structure, and a modified competitive ELISA method for quantification of BPA was developed. This system was based on BHPVA-BSA for polyclonal antibody production against bisphenolic structure, and BHPVA-HRP for determination of BPA substituting detection antibody in competitive reaction. Recovery was assessed at 10 different concentrations (2-1000 ng/ml) of BHPVA, and the recovery range was from 96.3% to 107.2%. The variation was from 6.2% to 9.8% for intra assay and from 10.1% to 12.6% for inter assay. The quadratic was used to establish the curve regression. The range was found to be between 2 and 1000 ng/ml. This modified competitive ELISA method has proven to be a very useful tool for quantification of BPA without the unexpected interaction of BSA and anti-BSA polyclonal antibody.

To observe the effects of acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) on distribution taxis of paclitaxel in mice with lung cancer to discuss targeted relationship between acupoints and corresponding viscera.According to randomized digital table, 315 SPF-grade BALB/C female mice were divided into 7 groups: blank group (group A), model group (group B), medication group (group C), acupuncture at non-acupoint group (group D), acupuncture at Feishu group (group E), acupuncture at Lingtai group (group F) and acupuncture at Feishu and Lingtai group (group G), 45 mice in each one. Except the blank group, the remaining groups were treated with N-nitroso-tris-chloroethyl urea (NTCU) to establish the model of squamous-cell carcinoma. After model establishment, group A, group B and group C were not treated with acupuncture; group A and group B were treated with intraperitoneal injection of 0.9% sodium chlorvde solution by 6 mL/kg while group C was treated with intraperitoneal injection of paclitaxel by 6 mL/kg. The group D, group E, group F and group G were treated with acupuncture at non-acupoint, "Feishu" (BL 13), "Lingtai" (GV 10) and "Feishu" (BL 13) plus "Lingtai" (GV 10), respectively, then were intraperitoneally injected with paclitaxel by 6 mL/kg. The treatment was all given once a day for continuous 10 days. 15 min, 30 min, 1 h, 2 h, 8 h, 12 h and 24 h after the treatments, 6 mice in each group were randomly selected and sacrificed to collect samples of lung, liver, spleen, kidney and heart, etc. High performance liquid chromatography was applied to measure the concentration of paclitaxel in each organ (lung, liver, spleen, kidney and heart) at different time points.(1) The content of paclitaxel in lung, kidney and heart reached the peak at 2 h, then decreased significantly in group C, group D, group E, group F and group G; the content of paclitaxel in spleen showed downtrend at each time point. The content of paclitaxel in liver reached the peak at 2 h in group C and group D; the content of paclitaxel reached the peak at 8 h in group E, group F and group G. (2) The content of paclitaxel in lung in group E and group G was higher than that in group C and group D at each time point (all P0.01); the content of paclitaxel in lung in group F was higher than that in group C (P0.01) and group D (P0.01) only at time point of 2 h. The content of paclitaxel in lung in group G was higher than that in group F at each time point (all P0.05). There was no statistical difference between group G and group E (all P0.05).Acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) could influ- ence the metabolism of paclitaxel in lung-cancer mice, leading to distribution change in each organ. As a result, it could cause targeting effects, which is more significant at "Feishu" (BL 13) and "Lingtai" (GV 10).

A sandwich enzyme-linked immunosorbant assay (ELISA) was developed to quantitatively detect Bullfrog (Rana catesbeiana) vitellogenin (VTG) levels. This procedure involved a sandwich ELISA using monoclonal antibodies of BVmA1 and BVmA2 against Bullfrog-VTG, and BVmA1 conjugated to horseradish peroxidase as the detection antibody. The assay range was between 9.4 ng/ml and 1200 ng/ml and the recovery of the VTG added to Bullfrog control male serum was 92.0-108.8%. Male Bullfrog was induced by injection 17beta-estradiol (E2) for four weeks and Bullfrog-VTG levels were measured each week. Histological analysis was performed for investigating the correlation of the effect to male reproduction and Bullfrog-VTG level variation depending on E2 dose. After two weeks of E2 exposure, the induced Bullfrog-VTG level was significantly higher than Bullfrog control female (p0.05). After four weeks of E2 exposure, the rupture and fusion of seminiferous tubule in the testes of male Bullfrog were shown and provided direct evidence that the reproduction of male Bullfrog was affected by estrogenic compounds. Bullfrog-VTG bioassay, using the sandwich ELISA, could be a sensitive and useful tool for quantification of estrogenic principles in aquatic environments.

This study explored the mechanism of electro-acupuncture (EA) at PC6 to improve the heart function by regulating the cardiac transient outward potassium current (=Ito) channel in myocardial ischemia (MI). Kv1.4 is the main component of the slow Ito (Ito. s) channel. Kv4.2 and Kv4.3 are the main components of the fast Ito (Ito. f) channel. KChIP2 is a compound protein of Ito channel. In this experiment, MI was induced by injecting isoproterenol in rats, and the gene and protein expressions of Kv1.4, Kv4.2, Kv4.3 and KChIP2 were lower [The gene and protein decreased (42.8±4.3)% and (37.2±4.7)% respectively.] than the control rats significantly (p < 0.05). After MI induction, the MI rats were divided into three groups, including PC6, LU7 (Lieque-point) and Non-acupoint group which were acupunctured at once a day for 7 days respectively. After EA at PC6 and LU7, the gene and protein expressions showed obvious increased [EA at PC6: the gene and protein increased (50.2±5.4)% and (39.1±5.3)% respectively. EA at LU7: the gene and protein increased (24.6±2.8)% and (22.0±5.5)% respectively] and they are all higher than MI rats significantly (p < 0.05). EA at PC6 and LU7 showed the same variation, and the effects of EA at PC6 and LU7 were better than Non-acupuncture-point (p < 0.05). The effects of EA at PC6 were significantly better [The gene and protein increased (19.7±2.7)% and (14.1±4.4)% respectively] than LU7 (p < 0.05). PC6 is an acupoint of the pericardium meridian, and the pericardium meridian which corresponds to adrenal gland according to Omura Y's research, can affect the heart function directly. LU7 belong to the lung meridian, and the lung plays an important factor in blood circulation according to TCM. So PC6 is more effective than LU7 on heart function regulation. The results suggested that PC6 showed the target effect of meridian specificity on regulating the Kv channel in MI.

There were applications of eye acupuncture for stroke patients. Unfortunately, similar to many other Traditional Chinese Medicine (TCM) treatments, it lacks comprehensive evaluation and system review for its effect and safety.Objective. This study is a systematic review to appraise the safety and effectiveness of eye acupuncture for stroke.Methods. “Eye acupuncture therapy” in eleven databases was searched by randomized controlled trials and quasi-randomized controlled trials. The search activity was ended in April 2014. The data were extracted and assessed by three independent authors. Rev Man 5.0 software was used for data analysis with effect estimate presented as relative risk (RR) and mean difference (MD) with a 95% confidence interval.Results. Sixteen trials (1120 patients) were involved with generally poor methodological quality. The study indicated that when eye acupuncture was combined with western medicine compared to western medicine, there was a significant difference in the areas of mental state, swallow function, and NDS. When eye acupuncture was combined with western medicine and rehabilitation compared to western medicine and rehabilitation, there was significant difference in the changes of SSS, FMA, and constipation symptoms evaluation. No adverse events or side effects have been reported.Conclusions. The current evidence is insufficient and the rigorously designed trials are warranted.

To study whether acupuncture affects the tissue distribution of Paclitaxel in mouse lung carcinoma.Totally 90 mice were divided into Paclitaxel group, Paclitaxel + Feishu (BL13) group, and Paclitaxel + Lingtai (DU10) group. Each group was consisted of 30 mice. After Paclitaxel injection, the mice received electro-acupuncture at Feishu or Lingtai acupoints once a day for 8 days. The effect of acupuncture on the tissue distribution of Paclitaxel was evaluated using high-performance liquid chromatography at 1, 2, and 3 h, respectively. The lung, liver, spleen, and kidney were examined for the concentration of Paclitaxel seperately.Paclitaxel was widely distributed in various organs, particularly in the lung, liver, and kidney. Acupuncture at Lingtai or Feishu acupoints resulted in an obvious decrease of Paclitaxel distribution in kidney and delayed Paclitaxel distribution in liver. Meanwhile, it increased the time of metabolism. Acupuncture at Feishu acupoint facilitated the delivery of Paclitaxel to lung more effectively than did acupuncture at Lingtai acupoint.Applying acupuncture at particular acupoints can influence tissue distribution of Paclitaxel. Tissue distribution change might be one of the mechanisms of acupuncture treatment during chemotherapy.

Based on traditional acupuncture-moxibustion treatment ideas, with differentiation of channels and collaterals as main part and feature, the important role of associated symptom and sign-based acupoint selection in acupuncture-moxibustion treatment is explained from angles of philosophy and medicine. Combined with clinical experience of acupuncture and moxibustion, the category of associated symptom and sign-based acupoint selection is explained in detail to make sure the accuracy of acupuncture-moxibustion differentiation. It could show uniqueness and advantage of theory and clinic in acupuncture-moxibustion and provide theoretical references in making acupuncture-moxibustion prescription to improve effectiveness of clinical treatment.

This study investigated the propagated sensation along meridians (PSM) produced respectively by acupuncture at a specific acupoint of right-side Quchi (LI11), a nonacupoint on meridian (control meridian point), and neither meridian nor acupoint (control point). All the stimulated points were on the right brachioradialis along the large intestine meridian of hand Yangming. Surface electromyography (sEMG) was used to reflect the activity of the brachioradialis along the large intestine meridian of hand Yangming. The PSM rate of LI11 (59.21%) and the control meridian point (53.95%) were significantly higher than the control point (38.16%) (P<0.05). After acupuncture, the brachioradialis sEMG amplitude was5.08±2.93 uV at LI11,3.08±1.18 uV at the control point, and2.77±1.36 uV at the control meridian point. The amplitude of LI11 was significantly higher than both the control meridian point and the control point (P<0.05). When the sEMG activity of brachioradialis returned to the stable base line, brachioradialis sEMG duration at LI11 (265±87.87 s) was significantly longer than that at the control meridian point (91.69±42.98 s) and the control point (83.31±32.76 s) (P<0.05). In conclusion, acupuncture activated PSM at all points but showed an acupoint specificity at LI11 and a meridian specificity at the control meridian point.

Recent evidence suggests that chloride (CL−) channels are involved in myocardial ischemia. In this study, the impact of acupuncture on the protein expressions of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) and CLC-2CL−channel of the rats with myocardial ischemia were tested and its mechanism was explored. The rats for experiment were distributed randomly into 5 groups: blank control group, modeling control group, Neiguan (PC-6) treatment group, Lieque (LU-7) control group, and Non-acupoint control group. The rats of all groups, except the blank control group, had myocardial ischemia via multiple subcutaneous injection of isoproterenol (ISO). Electroacupuncture treatment was given to Neiguan (PC-6) treatment group, Lieque (LU-7) control group, and Non-acupoints control group, respectively, once a day for 7 days. The results show that acupuncture can alleviate the myocardial ischemia of cardiac tissue, decrease significantly the activities of serum SOD and MDA, and thereby influence the protein expressions of CFTR and CLC-2 inCL−channels. The results of the study implies that acupuncture suppresses the pathological changes of cardiac tissue of rats with myocardial ischemia and regulates the protein expression of CFTR and CLC-2CL−channels, which may serve as one possible mechanism to reduce myocardial ischemia.

To observed efficacy differences of acupuncture at "Zusanli" (ST 36) in rats with asthma and asthma with spleen-deficency, so as to investigate the therapeutic mechanism.Sixty SD rats were randomly divided into 5 groups according to their weight, named as an asthma with spleen-deficency group (group A), an acupuncture on asthma with spleen-deficency group (group B), an asthma group (group C), an acupuncture on asthma group (group D) and a control group. The rat models with spleen-deficiency in the first two groups were set up by TCM, then the rats of asthma model in the first four groups were induced by egg albumin, but the control group was treated by the same dose of saline. The group B and the group D were both treated with acupuncture at "Zusanli" (ST 36), once each day for 8 days, and the other groups remained unhandled. The mRNA expressions of Fas and Bcl-2 in the lung tissues were detected by hybridization in situ and apoptosis was detected by TUNEL (terminal dexynucleotidyl transferase-mediated dutp nick end labeling).Compared with the control group, in both the group A and the group C, the expression of Fas mRNA significantly decreased, but the expression of Bcl-2 mRNA significantly increased (all P0.01), and eosinophils (EOS) counts significantly increased, but EOS apoptosis rate significantly decreased (all P0.01). Compared with the group C, in the group A, the expressions of Fas mRNA significantly decreased, but the expressions of Bcl-2 mRNA and EOS counts significantly increased (all P0.01). At the same time, compared with the corresponding asthma groups, in both acupuncture groups, Fas mRNA expression obviously increased, Bcl-2 mRNA expression was significantly reduced (all P0.01), EOS counts remarkably decreased and EOS apoptosis rate significantly increased (all P0.01). There were no significant differences in the expressions of Fas mRNA and Bcl-2 mRNA between the two acupuncture groups (both P0.05), but compared with group B,in the group D, EOS counts significantly decreased and EOS apoptosis rate significantly increased (both P0.01).Acupuncture at "Zusanli" (ST 36) can regulate the disorders of Fas mRNA and Bcl-2 mRNA expression in the state of both asthma and asthma with spleen-deficency, promote EOS apoptosis so as to inhibit the development of inflammatory reaction of asthma, showing that acupuncture at "Zusanli" (ST 36) has certain advantages on regulation of related gene of EOS in asthma with spleen-deficency.

To explore the distribution regulars of proximal and distal focus of muscle meridian regions in knee osteoarthritis patients.Seven hundred and sixty-five knees were selected in 516 cases of knee osteoarthritis. Under the guidance of muscle meridian theory, with the anatomical features of muscle meridian focus, the frequency and the location where the proximal and distal focus of muscle meridian regions appeared were calculated by palpation.Of all the points, 11 835 points of proximal focus of muscle meridian regions were found out by palpation, and 9455 points of distal focus of muscle meridian regions were found out. The percentages of the frequency that the focus of muscle meridian of Foot-Yangming, Foot-Taiyang, Foot-Shaoyang and three foot Yin meridians appeared at proximal points of knee were 37.1% (4388/11 835), 34.9% (4127/11 835), 9.5% (1129/11 835) and 18.5% (2191/11 835) respectively; and the percentage of the frequency that the focus of muscle meridian appeared at distal points of knee were 24.7% (2333/9455), 25.2% (2380/9455), 28.5% (2700/9455) and 21.6% (2042/9455).The proximal and distal focus of muscle meridian in knee osteoarthritis patients are closely related with anatomy structure and biomechanical characteristics; the distribution regulars of focus of muscle meridians study provides evidence for the selection of effective treatment points from different clinical acupuncture therapies.

Through analysis on sequencing of meridians and their muscle regions, their pertaining organs, run ning courses, linking and indications described in Zubi Shiyimai (Eleven Meridian of Foot and Hand), Yinyang Shiyimai (Eleven Meridian of Yinyang), Lingshu: Jingmai (Miraculous Pivot: Meridian) and Lingshu: Jinjing (Miraculous Pivot: Muscle Meridian), it is found that most of the indications of acupuncture in ancient time are symptoms of the muscle regions. 62.59% points of the national standard acupoints location close to tender points of the muscle regions, which indicates that the origin of early acupoints are tender points along the running courses of the muscle regions. Thus, it is concluded that meridians and their muscle regions have the same origin, which provides new train of thinking for a better comprehension of origin of meridians and collaterals.

Ghrelin, a 28-amino acid brain-gut peptide expressed in periphery tissues and the central nervous system, has been demonstrated to increase insulin sensitivity in adipocytes. Recent data have indicated that insulin resistance exists in the brain and is related to Alzheimer's Disease (AD). The aim of this study was to investigate whether ghrelin increased high glucose-induced hippocampal neuron insulin sensitivity, and further modulated tau phosphorylation. Hippocampal neurons were cultured in concentrations of 25 mM and 75 mM glucose. The effect of ghrelin on hippocampal neuronal insulin sensitivity was detected by [(3)H]-2-deoxy-D-glucose uptake. The expression of Akt, glycogen synthase kinase-3beta (GSK-3beta) and tau phosphorylation was determined via Western blotting. Culturation in 75 mM glucose resulted in decreased neuronal glucose uptake and an increase in the level of tau phosphorylation at Ser 199. In neurons treated with ghrelin for 1 h, neuronal glucose uptake was increased and tau hyperphosphorylation was improved. Ghrelin activated Akt and GSK-3beta phosphorylation, whereas phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin eliminated ghrelin's effect on neuronal glucose uptake and tau phosphorylation. This study demonstrated that ghrelin increased insulin-stimulated neuronal glucose uptake in 25 mM or 75 mM glucose, raised insulin sensitivity, improved insulin resistance and decreased tau abnormal phosphorylation via the PI3-K/Akt-GSK pathway. Ghrelin is a potential new medicine in the treatment of AD.

Heat shock factor 1 (HSF1) primarily regulates various cellular stress responses. The role of alpha-helix1 (H1) in its DNA-binding domain (DBD) during HSF1 activation remains unknown. Here, HSF1 lacking H1 loses its heat-induced activity, suggesting the importance of the latter. Furthermore, the CD spectra and AMBER prediction show that this H1 deficiency does not change the structure of HSF1 monomer, but does impact its heat-induced trimerization. Point mutation showed that Phe18 in H1 interacts with Tyr60, and that Trp23 interacts with Phe104 by an aromatic-aromatic interaction. Thus, the presence of H1 stabilizes the DBD structure, which facilitates the heat-induced trimerization and DNA-binding of HSF1.

Serum vitellogenin (VTG) contents of wild goldfish (Carassius auratus) were investigated as a sensitive biomarker for artificial estrogenic compounds in aquatic environments. Goldfish was sampled from a pristine area, a river situated 5 km downstream from a sewage treatment works (STW), and also from the Young-San River in Korea. The female yolk precursor protein VTG was not detected when gonadosomatic index (GSI) was less than 0.85%, while VTG levels of10 microg/ml were found in males whose GSI was less than 1.53%. In male goldfish sampled from STW and the Young-San River, the higher VTG corresponded to lower GSI. This study suggested a trend that gonad development was connected to VTG levels in both sexes, and the application of GSI and histological analysis provide an attractive possibility that it could be included in the panel of markers used for estrogenic activity investigation of aquatic environments.

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantitatively detect Carassius auratus vitellogenin (VTG) levels. The protein levels in fish plasma are useful aquatic biomarkers of estrogenic compounds. This procedure involved an ELISA using monoclonal antibodies of CVmA2 and CVmA7 against Carassius auratus VTG, and CVmA7 conjugated to horseradish peroxidase as the detection antibody. The assay range was between 1 and 401.5 ng/ml and the recovery of the VTG added to Carassius auratus plasma was 92.5-109%. An in vitro assay was performed to measure low levels of the VTG, using primary hepatocytes of Carassius auratus induced by 17-beta estradiol (E2). The detection limit was 1 ng/ml and 137 ng/ml at the maximum. Within each sex of wild Carassius auratus, VTG levels from the river next to sewage treatment works (STWs) were much higher than those from the feeding stream. The Carassius auratus VTG bioassay could be a sensitive and useful tool for quantification of estrogenic principles in aquatic environments.

Department of Chemistry and Center for Innovative Bio·Physio Sensor Technology, Department of Molecular Biology, Pusan National University, Busan 609-735, Korea. E-mail: jaspark@pusan.ac.kr Department of Bioengineering and Biotechnology, Pukyong National University, Busan 608-737, Korea Department of Biomedicinal Chemistry, and Institute of Functional Materials, Inje University, Kimhae 621-749, Korea Received May 28, 2007

Eukaryotic replication protein A (RPA) is comprised of a three subunits (70, 32, 14 kDa; p70, p32, p14 respectively). RPA has been identified as a human protein necessary for SV40 DNA replication in vitro and acting as a singlestrand DNA (ssDNA) binding protein with multiple functions in DNA replication, repair, and genetic recombination. Recent studies suggest that RPA functions are regulated in response DNA damage and that its ssDNA binding activity may be involved in this regulatory event. In our previous study, we found that RPA’s ssDNA binding activity is regulated by redox through cysteins in the putative zinc finger domain of the p70 subunit. A number of DNAbinding proteins have been identified in which their DNA binding activity is regulated by redox, although the regulatory role of the zinc finger domain is not clear. RPA70 has an evolutionarily conserved 4 cystein in the zinc-finger domain at 473 through 503 amino acids. Even though previous studies showed that the zinc-finger domain does not participate in DNA binding directly, it is still possible that the zinc-finger domain regulates DNA binding activity. To verify this possibility, we built mutant RPA (ZFD4) by substitution four cysteins for alanine and checked the DNA binding activity. The ZFD4 formed a stable complex with the ssDNA, even under oxidized conditions, and the amounts of DTT had no effect on its binding property (Figure 1a). The ZFD4 also showed strong binding activity in the non-reducing condition without DTT. The addition of H2O2 did not have much of effect on RPA-DNA complex formation (Figure 1a), whereas the wild-type RPA had H2O2 dependent activity. 7-9 These results suggest that the zinc-finger domain could regulate the ssDNA binding activity of RPA and that the cystein residues of the p70 subunit are essential in this process. We examined the effect of phosphorylation on ZFD4’s in regulation of ssDNA binding activity of RPA in the various redox conditions. The results showed phosphorylation of ZFD4 does not affect its DNA binding activity and suggests that phosphorylation could occur at other sites of the RPA complex (Figure 1a). In an effort to understand RPA’s regulatory function, we examined the structural changes of hRPA by the ssDNA binding using protease sensitive digestions. We examined