Effects of electroacupuncture on liver function in mice with chronic alcoholic liver injury: visual display by in vivo fluorescence imaging.

OBJECTIVE: Acupuncture can improve the symptoms of alcohol-induced bodily injury and has been accepted by the World Health Organization. In this study, in vivo fluorescence imaging (IVFI) was applied to display and evaluate the effect of electroacupuncture (EA) on liver function (LF) in mice with chronic alcoholic liver injury (cALI). METHODS: IVFI of the Cy5.5-galactosylated polylysine (Cy5.5-GP) probe targeting the liver asialoglycoprotein receptor (ASGPR) and liver indocyanine green (ICG) clearance was performed to visually evaluate the effect of EA at ST36 and BL18 on liver reserve function and hepatic metabolism in mice with cALI. In addition, changes in ASGPR expression, serum indexes of LF, and hepatic morphology were observed. RESULTS: After EA at ST36 and BL18, the ASGPR-targeted fluorescence signals (FS) in the liver increased significantly in cALI mice (p < 0.05) and exhibited relationships with liver ASGPR expression, liver ICG clearance, liver histology, and serum marker levels of LF in cALI mice undergoing EA intervention. CONCLUSIONS: As displayed by IVFI, EA at ST36 and BL18 appears to improve liver reserve function and inhibit the development of liver injury in mice with cALI. EA may have potential as a treatment strategy to protect against ALI.

[Sixty years of meridian-collateral exploration: a road of interdisciplinary research].

The theory of traditional Chinese medicine believes that meridian-collateral (jingluo) is an important system to maintain life and participates in the formation and cure of disease. Exploring the biological connotation of meridian-collateral is the key content in the modern research of traditional Chinese meridian. For more than 60 years, scholars at home and abroad have carried out the researches of meridian-collateral from the fields of biomedicine (e.g. physiology, anatomy and histomorphology), imaging, physics, chemistry, computer technology, etc. Meridian-collateral research in modern times demonstrates a multi-disciplinary historical development and provides multi-perspective scientific information for understanding meridian-collateral. The meridian-collateral research in future needs more accurate multidisciplinary intersection, which will be the light of hope to reveal the biological connotation of meridian-collateral.

[Effect of electroacupuncture on the hemorheology and hepatic microcirculation in mice with chronic alcoholic liver injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the hemorheology and hepatic microcirculation in mice with chronic alcoholic liver injury (cALI), so as to explore the microcirculation mechanism of EA underlying regulating liver function in cALI mice. METHODS: Forty Kunming mice were randomly assigned to control, model, acupoint EA and non-acupoint EA groups, with 10 mice in each group. The cALI model was established by gavage with 50% ethanol (15 mL/kg，2 times per day with 8 hours' interval) for 28 d. Mice in the acupoint EA group received EA at bilateral "Zusanli" (ST36) and "Ganshu"(GB11) for 20 min, once daily for 14 d. And mice in the non-acupoint EA group received EA the spot about 1 cm lateral to GB11 and 0.5 cm lateral to ST36 respectively for 20 min, once daily for 14 d. The hepatic blood perfusion (HBP) of mice was detected by laser speckle perfusion imaging. The index of hemorheology, serum markers of liver function and hepatic histology in mice were observed by automatic blood rheometer, automatic bio-analysis machine and H.E. staining, respectively. RESULTS: Compared with the control group, the HBP of the model group was significantly decreased (P<0.05); the low-cut whole blood viscosity, plasma viscosity, and erythrocyte sedimentation level were significantly increased （P<0.05）, and the red blood cell deformation index was significantly decreased (P<0.05); serum alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) increased significantly (P<0.05) in the model group. Hepatocyte steatosis accompanied by inflammatory cell infiltration and focal necrosis was observed in the model group. After EA at ST36 and GB11, and in comparison with the model group showed that the HBP was significantly increased (P<0.05); the low-cut whole blood viscosity, plasma viscosity, erythrocyte sedimentation rate and serum GGT were significantly decreased (P<0.05), and the red blood cell deformation index was significantly increased (P<0.05）; the degree of fatty degeneration in the liver tissue was reduced, and the focal necrosis was reduced. The plasma viscosity, erythrocyte sedimentation rate were significantly decreased (P<0.05), and the red blood cell deformation index was significantly increased in the non-acupoint EA group relevant to the model group (P<0.05). The erythrocyte sedimentation rate and the red blood cell deformation index of the acupoint EA group was significantly lower than that of the non-acupoint EA group (P<0.05). CONCLUSION: EA at ST36 and GB11 could improve the hemorheological disorder and the hepatic microcirculation, and inhibit the deve-lopment of liver injury in cALI mice.

[Analysis on the defensive function of defensive qi in Huangdi Neijing (The Yellow Emperor's Inner Classic)].

The defensive function of defensive qi in Huangdi Neijing (The Yellow Emperor 's Inner Classic) includes the effect of immune system of human body against the pathogen invasion. According to the basic research, it is believed that the circulation of defensive qi in meridian is interpreted as the flow of interstitial fluid in the low hydraulic resistance channel that is overlapped with lymphatic vessels, transporting the antigen information from interstitium to lymph nodes and generating adaptive immune response. Enhancing meridian circulation and promoting the flow of interstitial fluid may improve the immunity, which demonstrating the thought of traditional Chinese meridian as "pathogens failing to invade the body when the anti-pathogenic qi is strong enough inside".

[Display of low hydraulic resistance channels along Conception Vessel by in vivo fluorography in rats].

OBJECTIVE: To display the low hydraulic resistance channel along the running course of the Conception Vessel (CV) by using in vivo fluorography, so as to provide evidence for the existence of meridians of traditional Chinese medicine from the aspect of interstitial fluid channel. METHODS: A total of 24 male SD rats were randomly divided into 3 groups, namely CV-low impedance spots (CV-LIS), CV-low hydraulic resistance points (CV-LHRP) and non-meridian points (NMP) groups (n＝8 in each group). The CV-LIS (the imaged line between the xiphoid process and symphysis pubis), CV-LHRP (lateral and parallel to LIS) and NMP (lateral and parallel to CV-LHRP) were detected by using meridian detector and biofluid resistance meter, respectively. Sodium fluorescein solution (1%) was injected into the spot (A-spot, upper 1/5 of the CV-LIS), and one of the NMP (about 1 cm lateral and parallel to the A-spot, and the mid-point between the Kidney and Stomach meridian running courses) by using a microsyringe for observing the migration trace of sodium fluorescein. RESULTS: After injection of fluorescein, the migration trace of fluorescein appeared along the CV-LIS and CV-LHRP lines, and only local diffusion was found in the NMP group. The migration trace lengths along the CV-LIS and CV-LHRP were significantly longer than that of the NMP group (P<0.01), and the migration width of CV-LIS and CV-LHRP groups was significantly smaller than that of the NMP group (P<0.01). The migration speed of CV-LIS and CV-LHRP groups was significantly higher than that of the NMP group (P<0.01). CONCLUSION: Fluorography is able to be used to display the running course of CV, possibly supporting the existence of meridians from the interstitial fluid channel and providing a new method for visualization of meridians.

Analysis of the Spectral Characteristics of Pure Moxa Stick Burning by Hyperspectral Imaging and Fourier Transform Infrared Spectroscopy.

The objective of this study was to investigate the spectra characteristics (SC) at wavelengths of 400~1000 nm and 2.5~15.5 µm of pure moxa stick (MS) during its 25-minute burning process using new spectral imaging techniques. Spectral images were collected for the burning pure MS at 5, 10, 15, 20, and 25 min using hyperspectral imaging (HSI) and Fourier transform infrared spectroscopy (FTIR) for the first time. The results showed that, at wavelengths of 400~1000 nm, the spectral range of the cross section of MS burning was 750~980 nm; the peak position was 860 nm. At wavelengths of 2.5~15.5 µm, the spectral range of the cross section of MS burning was 3.0~4.0 µm; the peak position was approximately 3.5 µm. The radiation spectra of MS burning include litter red and amount of infrared (but mainly near infrared) wavelengths. The temperature, blood perfusion, and oxygen saturation increase of Shenshu (BL23) after moxibustion radiation were observed too. According to mechanism of photobiological effects and moxibustion biological effects, it was inferred that moxibustion effects should be linked with moxibustion SC. This study provided new data and means for physical properties of moxibustion research.

Evaluation of liver function and electroacupuncture efficacy of animals with alcoholic liver injury by the novel imaging methods.

Imaging methods to evaluate hepatic microcirculation (HM) and liver function (LF) by directly monitoring overall liver tissue remain lacking. This study establish imaging methods for LF that combines Laser speckle perfusion imaging (LSPI) and in vivo optical imaging (IVOI) technologies to investigate changes of hepatic microcirculation and reserve function in the animals gavaged with 50% ethanol (15 ml/kg·bw) for a model of acute alcoholic liver injury (ALI), and for evaluation of electroacupuncture (EA) effect. The liver blood perfusion and indocyanine green (ICG) distribution were observe by LSPI and IVOI separately. After EA, the livers were collected to measure the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), thromboxane A (TXA2), prostacyclin (PGI2) and endothelin (ET). The acquisitions of newly established LSPI of liver and ICG in vivo fluorescence imaging (ICG-IVFI), combining the results of other indexes showed: hepatic microcirculation perfusion (HMP) significantly reduced, ICG metabolism reduced, and ALT/AST increased in animal model with acute ALI. EA can reverse these changes. The use of LSPI of liver and ICG-IVFI, which was novel imaging methods for LF established in this study, could display the LF characteristics of ALI and the EA efficacy.

Effects of Electroacupuncture Stimulation at "Zusanli" Acupoint on Hepatic NO Release and Blood Perfusion in Mice.

The study is to observe the influence of electroacupuncture (EA) stimulation at "Zusanli" (ST36) on the release of nitric oxide (NO) and blood perfusion (BP) in the liver and further explore whether the hepatic blood perfusion (HBP) changes were regulated by EA ST36 induced NO in nitric oxide synthase inhibited mice. The HBP change of the mice was detected by laser speckle perfusion imaging (LSPI) before and after being given interventions, and the NO in liver tissue was detected by nitric acid reductase in each group. The NO levels and HBP in the L-NAME group were significantly lower than those in the control group (P < 0.01). The NO level and HBP increase in EA group were significantly higher than those in control group (P < 0.05). The NO level in the L-NAME EA group was slightly higher than that in the L-NAME group. The HBP increase in the L-NAME EA group was not statistically significant. These results showed that EA could accelerate the synthesis of NO and thereby increase HBP via vasodilation in liver tissue.

A pilot study on effects of acupuncture and moxibustion by hyperspectral imaging technique.

This study was to observe the effects of acupuncture and moxibustion on spectrum features of acupoint using hyperspectral imaging (HSI) technique. HSI of the Neiguan (PC6) in the acupuncture groups, moxibustion groups, and control groups was scanned by the hyperspectral imager to analyze the spectrum features and the variations within the wavelength of 400-1000 nm and explore the relationship between the spectral characteristics and effects of acupuncture and moxibustion. The light absorption intensity was slightly reduced within the wave band of 540-590 nm after acupuncture. The absorption intensity of PC6 before moxibustion was significantly higher than that after moxibustion, and the maximum reduction was found at the wavelength of 580 nm with 20.5% reduction, P < 0.05. There was no significant change of the spectrum of palm and PC6 and the spectrum curves of the acupoint were basically identical in control group. The light absorption intensity of PC6 of human body was weakened after Acu-mox. Specific wavelengths were all exhibited at 580 nm and the effect of moxibustion was more significant. HSI technique can be used to measure the spectral characteristics of the acupoint areas. This first time research would be significant and beneficial for study on the effect of acupuncture and moxibustion.

Investigation of hepatic blood perfusion by laser speckle imaging and changes of hepatic vasoactive substances in mice after electroacupuncture.

The study was conducted to observe the effect of electroacupuncture (EA) on hepatic blood perfusion (HBP) and vascular regulation. We investigated 60 male anesthetized mice under the following 3 conditions: without EA stimulation (control group); EA stimulation at Zusanli (ST36 group); EA stimulation at nonacupoint (NA group) during 30 min. The HBP was measured using the laser speckle perfusion imaging (LSPI). The level of nitric oxide (NO), endothelin-1 (ET-1), and noradrenaline (NE) in liver tissue was detected by biochemical methods. Results were as follows. At each time point, HBP increase in ST36 group was higher than that in the NA group in anesthetized mice. HBP gradually decreased during 30 min in control group. The level of NO in ST36 group was higher than that in NA group. The level of both ET-1 and NE was the highest in control group, followed by NA group and ST36 group. It is concluded that EA at ST36 could increase HBP possibly by increasing the blood flow velocity (BFV), changing vascular activity, increasing the level of NO, and inhibiting the level of ET-1 in liver tissue.

[Effects of electroacupuncture stimulation of "Zusanli" (ST 36) on the regional vascular intercellular Adhesion Molecule-1 mRNA expression and mast cell activity in normal rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Zusanli" (ST 36) on intercellular adhesion molecule-1 (ICAM-1) mRNA expression and mast cell (MC) distribution in the stimulated subcutaneous tissues in nor- mal rats, so as to study the correlation between ICAM-1 mRNA expression and MC activity during EA. METHODS: Twenty Wistar rats were randomly assigned to normal group and EA group. EA(2 Hz/15 Hz, 0. 1 mA) stimulation was applied to the bilateral "Zusanli" (ST 36) for 25 min. The expression of OICAM-1 mRNA in subcutaneous tissues of the left "Zusanli" (ST 36) area was detected by in situ hybridization, and MC activity of the same tissues was observed under microscope after staining with acetylcho- line esterase (AChE) and toluidine blue (TB, counterstaining), respectively. RESULTS: Compared with the normal group, the ICAM-1 mRNA expressed at a markedly higher level in the subcutaneous tissues of the "Zusanli" (ST 36) region in the EA group (P<0. 01), and MC counts were also significantly increased in the EA group (P<0. 01). CONCLUSION: EA stimulation up-regulates the expression of ICAM-1 mRNA and enhances MC release from the stimulated regional subcutaneous tissue in healthy rats, suggesting ICAM-1 played an important role in affecting the movement and recruitment of MCs towards acupoint during EA.

[Blood perfusion in different facial acupoint areas and its changes after acupuncture stimulation of Hegu (LI 4) displayed by laser Doppler imager in healthy volunteers].

OBJECTIVE: To observe the blood perfusion levels in different facial acupoint regions and changes after acupuncture stimulation of Hegu (LI 4). METHODS: Thirty healthy volunteer subjects were equally divided into control group and acupuncture group. Blood perfusion levels of different facial acupoint areas were detected using a Laser Doppler Imager. Acupuncture stimulation was applied to bilateral Hegu (LI 4) by using two filiform needles which were manipulated for a while till "Deqi", followed by keeping them in position for 30 min. The needles were then manipulated once again every 10 min. RESULTS: The blood perfusion levels of facial acupoint regions were from 0.73 to 1.17 PU in healthy volunteers, with relatively higher levels being in Juliao (ST 3), Kouheliao (LI 19), Sibai (ST 2) and Chengqi (ST 1) regions and relatively lower levels in Xiaguan (ST 7), Qianzheng (Ex-HN) and Sizhukong (TE 23) regions, but without significant difference between the isonym acupoint areas of the bilateral face in the control group. Following acupuncture stimulation of bilateral Hegu (LI 4), the blood perfusion volumes were remarkably increased in the acupoint regions on bilateral sides of the face (P < 0.05, P < 0.01). CONCLUSION: Acupuncture of Hegu (LI 4) can effectively increase blood perfusion volume of the acupoint areas of the bilateral face in healthy volunteers.

[Effect of acupoint-injection of stem cell factor antibody on electroacupuncture induced changes of count and degranulation of subcutaneous mast cells in the local region in rats].

OBJECTIVE: To observe the alternations of local mast cells (MCs) in the number and functional activities after electroacupuncture (EA) and injection of stem cell factor (SOF) antibody into "Weishu" (BL 21) and "Zusanli" (ST 36) in rats. METHODS: Thirty Wistar rats were randomly assigned to normal, EA, and EA + SCF antibody injection (EA-ABI) groups (n=10). EA was applied to the left "Weishu"(BL 21) and "Zusanli" (ST 36) for 25 min. For rats of EA+ ABI group, SOF-AB (1:200, 0.1 mL) was respectively injected into BL 21 and ST 36, following by EA. The tissues around the two acupoints areas were sampled to be stained with acetylcholine esterase (AchE) and toluidine blue (TB), respectively. The counts and degranulation ratio of MOs in the acupoint region were recorded and observed under microscope. RESULTS: In comparison with the normal group, MCs count was increased significantly in the EA of "Weishu" (BL 21) group (P < 0.05), mildly in the EA of "Zusanli" (ST 36) group (P > 0.05), and downregulated considerably in both EA-ABI-BL 21 and EA-ABI-ST 36 groups (P < 0.05) . Compared with the normal group, the degranulation ratio of MCs was increased significantly in EA-BL 21, EA-ST 36, EA-ABI-BL 21 and EA-ABI-ST 36 groups (P < 0.5). No significant differences were found between EA-BL 21 and EA-ABI-BL 21 groups, and between EA-ST 36 and EA-ABI-ST 36 groups in the degranulation ratios (P > 0.05). CONCLUSION: Local injection of SCF antibody in BL 21 and ST 36 can significantly suppress EA-induced increase of the number and degranulation ratio of local subcutaneous MCs in rats, suggesting that SCF is an important factor affecting the activity and recruitment of MCs around the acupoint during EA.

[Study on the manifestation of facial infrared thermography induced by acupuncturing Guangming (GB 37) and Hegu (LI 4)].

OBJECTIVE: To manifest the specificity of acupoints objectively through observing the relationship of both Guangming (GB 37) and Hegu (LI 4) with the some areas of face. METHODS: The patients with peripheral facial paralysis were divided into the Guangming (GB 37) group and Hegu (LI 4) group, and punctured respectively. Before and after treatment, the distribution and change of facial temperatures were measured with infrared thermography and the data on each area was comparatively analyzed. RESULTS: After punctured at Guangming (GB 37), the temperature went up in the areas around the eyes of both health and affected sides, especially on the affected side, which was significantly different from the other facial areas (all P<0.05). After punctured at Hegu (IA 4), the temperature obviously went up in the area around the mouth, which was significantly different from other areas of face, such as the areas of Yintag and health eye. The temperature in the area around the affected eye was significantly higher than that of the area around the health eye. CONCLUSION: There is some close coordination between Guangming (GB 37) and eye area, as well as between Hegu (LI 4) and mouth area, suggesting the specificity of acupoints.