OBJECTIVE: To study the regularity of central response to thermal needle stimulation of \"Zusanli\" (ST36) at different temperature, and to analyze the temperature difference of central responses.
METHODS: Six male C57BL/6j adult mice were used in the present study. For observing activities of neurons in the hindlimb region of left primary somatosensory cortex (S1HL, A/P=0.46 mm, M/L=1.32 mm, D/V=-0.14 mm) by using a fast high-resolution miniature two-photon microscopy (FHIRM-TPM), the mice were anesthetized with 3% isoflurane (inhalation), with its head fixed in a stereotaxic apparatus, then, adeno-associated virus (AAV-hSyn-GCaMP6f-WPRE-hGHpA, for showing intracellular calcium transients in neurons transfected) was injected into the left S1HL region using a micro-syringe after scalp surgical operation. The mice\'s right ST36 were stimulated using internal thermal needles with the temperature being 43 ℃, or 45 ℃, or 47 ℃, separately. Image J software and MATLAB 2020b software were used to process the image data of neuronal calcium activity (Ca2+ signaling) in the left S1HL region, including the instant maximum calcium peak value (ΔF/F) in 2 s, instant calcium spike frequency in 2 s, short-term calcium peak value (ΔF/F) in 3.5 min, short-term calcium spike frequency in 3.5 min, calcium peak duration in 3.5 min, maximum calcium peak value (ΔF/F) at the 1st , 2nd and 3rd min, and calcium spike frequency at the 1st, 2nd and 3rd min after thermal needle stimulation.
RESULTS: In comparison with the normal temperature needle stimulation, the instant intracellular maximum calcium peak value, instant calcium spike frequency, short-term maximum calcium peak value, short-term calcium spike frequency, and calcium peak duration of S1HL neurons in response to 43 ℃, 45 ℃ and 47 ℃ internal thermal needle stimulation of ST36 were significantly increased (P<0.001, P<0.01). Comparison among the 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation showed that the 45 ℃ thermal needle stimulation was obviously superior to 43 ℃ and 47 ℃ thermal needle stimulation in increasing instant calcium spike frequency, short-term calcium spike frequency and calcium peak duration of S1HL neurons (P<0.001, P<0.01). The 47 ℃ thermal needle stimulation was stronger than 43 ℃ and 45 ℃ thermal needle stimulation in increasing the instant maximum calcium peak value (P<0.001). The maximum calcium peak value was apparently higher (P<0.001) at the 2nd min than that at the 1st and 3rd min after 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation. No significant differences were found in the short-term maximum calcium peak value among the 3 thermal needle stimulation and in the calcium spike frequency among the 3 time points after 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation.
CONCLUSIONS: S1HL neurons respond to all 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation of ST36 in mice, while more actively to 45 ℃ thermal needle stimulation.
目的: 研究不同温度热刺激“足三里”的中枢响应规律，分析中枢响应的温度差异性。方法: 健康成年C57BL/6j小鼠6只，采用动物脑立体定位仪定位小鼠大脑左侧初级体感皮层后肢区（S1HL），注射0.3 μL腺相关病毒，通过双光子显微镜观察其神经元钙活动荧光成像，以荧光强度显示神经元中的钙离子浓度。分别以43、45、47 ℃热刺激小鼠右侧“足三里”，使用双光子显微镜直接记录小鼠大脑左侧S1HL神经元活动，观察并比较S1HL神经元对3种温度热刺激“足三里”的响应差异性，包括热刺激后2 s内的即时最大钙尖峰值和即时钙尖峰频率，3.5 min内的短时最大钙尖峰值、短时钙尖峰频率和钙尖峰持续时间，以及3 min内的分时段最大钙尖峰值和分时段钙尖峰频率。结果: 与常温比较，43、45、47 ℃热刺激“足三里”均可使小鼠左侧S1HL神经元的即时最大钙尖峰值、即时钙尖峰频率、短时最大钙尖峰值、短时钙尖峰频率和钙尖峰持续时间升高（P<0.001，P<0.01）。不同温度热刺激比较，从即时最大钙尖峰值来看，小鼠S1HL神经元对47 ℃热刺激响应程度最高（P<0.001）；从即时钙尖峰频率来看，其对45 ℃热刺激响应最频繁（P<0.001）。从短时最大钙尖峰值来看，不同温度热刺激对其影响差异无统计学意义；从短时最大钙尖峰频率来看，小鼠S1HL神经元对45 ℃热刺激响应最频繁（P<0.001）。从钙尖峰持续时间来看，小鼠S1HL神经元对45 ℃热刺激响应更为明显（P<0.01）。从神经元响应分时段分析来看，在3 min内，43、45、47 ℃热刺激下，小鼠S1HL神经元最大钙尖峰值都在第2分钟达到最高（P<0.001）；钙尖峰频率与热刺激时段无显著联系。结论: S1HL神经元对43、45、47 ℃热刺激“足三里”均有响应，且响应存在特定温度差异性，小鼠S1HL神经元对45 ℃热刺激“足三里”响应较为活跃。.

OBJECTIVE: To observe the effects of moxibustion at \"Zusanli\" (ST36) on the expression levels of tumor necrosis factor (TNF)-α, TNF receptor 1 (TNF-R1), p38 mitogen-activated protein kinase (P38 MAPK), and transient receptor potential vanilloid 1 (TRPV1) in the colon tissue of mice with chronic ulcerative colitis (UC), so as to explore the underlying mechanisms of moxibustion in improving visceral hypersensitivity in chronic UC.
METHODS: Male C57BL/6J mice were randomly divided into normal group, normal with moxibustion (NM) group, model group, and model with moxibustion (MM) group, with 10 mice in each group. The chronic UC model was established by drinking 2.5% dextran sodium sulfate for 3 cycles. Mice in the NM and MM groups received moxibustion at ST36 for 20 min, 5 days per week with a 2-day break, for a total of 4 weeks. The disease activity index (DAI) score of each group was evaluated before and after treatment. The minimum volume threshold of abdominal wall retraction reflex (AWR) was measured to observe the intestinal sensitivity of mice. The colon length was measured. The pathological changes of colon tissue were observed by HE staining. The expression of mucin in colon goblet cells was detected by periodate Scheff staining. The intestinal fibrosis was observed by Masson staining. The number of trypsin-positive cells (i.e., mast cell) and the expression level of TNF-α in colon tissue were detected by immunofluorescence staining. The expression levels of TNF-R1, P38 MAPK and TRPV1 in colon tissue were detected by immunohistochemistry.
RESULTS: Compared with the normal group after treatment, the model group showed increased DAI score (P<0.001), decreased AWR minimum volume threshold (P<0.01), shortened colon length (P<0.001), significant inflammatory infiltration in the colon tissue, reduced mucin secretion (P<0.01), increased collagen fiber deposition (P<0.001), and elevated expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). Compared with the model group, the MM group showed decreased DAI score (P<0.01), increased AWR minimum volume threshold (P<0.001), elongated colon length (P<0.001), reduced inflammatory cell infiltration, improved integrity of mucosal glandular structure, enhanced mucin secretion (P<0.01), decreased collagen fiber deposition (P<0.001), decreased number of mast cells in the colon tissue (P<0.001), and decreased expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). There were no significant differences in the above index between the NM group and the normal group.
CONCLUSIONS: Moxibustion can reduce visceral hypersensitivity, alleviate inflammatory infiltration and fibrotic damage in the colon tissue of mice with chronic UC. These effects may be associated with the down-regulation of TNF-α, TNF-R1, P38 MAPK, and TRPV1 expression in colon.
目的: 观察艾灸“足三里”对慢性溃疡性结肠炎（UC）小鼠结肠组织中肿瘤坏死因子-α（TNF-α）、肿瘤坏死因子受体1（TNF-R1）、P38丝裂原活化蛋白激酶（P38 MAPK）、瞬时感受器电位香草酸受体1（TRPV1）表达的影响，探讨艾灸改善慢性UC内脏高敏的可能机制。方法: 雄性C57BL/6J小鼠按体质量随机分为正常组、正常+艾灸组、模型组、模型+艾灸组，每组10只。以2.5%的葡聚糖硫酸钠循环3个周期饮用法建立慢性UC模型。正常+艾灸组和模型+艾灸组小鼠予以“足三里”艾灸，每次20 min，每周连续干预5 d，休息2 d，共4周。评价各组小鼠治疗前后的疾病活动指数（DAI）评分；测定各组小鼠腹壁回撤反射（AWR）最小容量阈值以观察小鼠肠道敏感性；测量各组小鼠结肠长度；HE染色观察各组小鼠结肠组织病理变化；过碘酸雪夫染色检测各组小鼠结肠组织杯状细胞中的黏液素分泌；Masson染色观察各组小鼠肠道纤维化情况；免疫荧光染色法检测各组小鼠结肠组织类胰蛋白酶阳性细胞数（即肥大细胞数）及TNF-α表达水平；免疫组织化学法检测各组小鼠结肠组织TNF-R1、P38 MAPK、TRPV1表达水平。结果: 艾灸治疗后，与正常组相比，模型组小鼠DAI评分升高（P<0.001），AWR最小容量阈值降低（P<0.01），结肠长度缩短（P<0.001），结肠组织炎性浸润明显，深入基底层，黏液素分泌减少（P<0.01），胶原纤维增加（P<0.001），结肠组织中肥大细胞数量增加（P<0.001），TNF-α、TNF-R1、P38 MAPK、TRPV1表达水平均明显升高（P<0.001，P<0.01，P<0.05）。与模型组相比，模型+艾灸组小鼠DAI评分降低（P<0.01），AWR最小容量阈值升高（P<0.001），结肠长度增加（P<0.001），炎性细胞浸润减少，黏膜肠腺结构完整性增加，黏液素分泌增多（P<0.01），胶原纤维减少（P<0.001），结肠组织中肥大细胞数量减少（P<0.001），TNF-α、TNF-R1、P38 MAPK、TRPV1表达水平均明显降低（P<0.001，P<0.01，P<0.05）。正常+艾灸组小鼠各项指标与正常组相比差异无统计学意义。结论: 艾灸可减轻慢性UC小鼠内脏高敏，减轻肠道肥大细胞炎性浸润及纤维化损伤，该效应可能与艾灸下调结肠TNF-α、TNF-R1、P38 MAPK、TRPV1表达有关。.

OBJECTIVE: To observe the effect of electroacupuncture (EA) at \"Zusanli\"(ST36) on intestinal mucosal damage, intestinal mucosal oxidative stress injury and apoptosis induced by 5-fluorouraeil (5-FU) chemotherapy in colorectal cancer-bearing mice.
METHODS: Thirty male BALB/c mice were randomly divided into normal control, colorectal cancer (CT26), 5-FU, non-acupoint and ST36 groups, with 6 mice in each group. Except for those of the normal control group, mice of the remaining 4 groups received subcutaneous implantation of colorectal CT26 cell suspension (0.1 mL) in the right armpit for establishing colorectal cancer model. Rats of the 5-FU group, non-acupoint group and ST36 group were given with 5 mg/mL 5-FU solution once every 3 days for a total of 21 days. For mice of the non-acupoint group and ST36 group, EA (2 Hz, 1-2 mA) was applied to bilateral ST36 or non-acupoints (the bilateral sunken spots about 3 mm to the midpoint between the tail root and the anus) for 5 min after each intraperitoneal infusion of 5-FU, once every 3 days, for a total of 21 days. After the intervention, the diarrhea index was assessed. The length of colon (from the endpoint of cecum to the anal orifice) was measured. Histopathological changes of colonic mucosa were observed by H.E. staining, and the length of colonic villi was measured. The content of malondialdehyde (MDA), and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of colonic tissue were detected by thibabituric acid, xanthine oxidase and colorimetric method, respectively. The rate of cell apoptosis in the colonic tissue was measured by TUNEL assay. The positive expressions of Bax and Bcl-2 in colonic tissue were determined by immunohistochemistry.
RESULTS: The CT26 model group didn\'t show any significant changes in the diarrhea index, colon length, colon villus length, MDA content, SOD and GSH-Px activities, colonic cell apoptosis rate, and Bax and Bcl-2 expression levels when compared with the normal group. Compared with the CT26 group, the 5-FU group had a remarkable increase in the diarrhea index, MDA content, colonic cell apoptosis rate and Bax expression level (P<0.01, P<0.05), and a marked decrease in the colon length, colon villus length, SOD and GSH-Px activities and Bcl-2 expression level (P<0.01), suggesting the side effects of administration of 5-FU. Compared with the 5-FU group, the diarrhea index, MDA content, colonic cell apoptosis rate and Bax expression level were markedly decreased (P<0.05, P<0.01) and those of the colon length, colon villus length, SOD and GSH-Px activities and Bcl-2 expression level were obviously increased (P<0.01) in the ST36 group. Compared with the 5-FU group, the non-acupoint group also had an increase in the colon villus length, SOD and GSH-Px activities (P<0.01, P<0.05) and a decrease in the cell apoptosis rate (P<0.01).
CONCLUSIONS: EA at ST36 has a positive effect in reducing intestinal mucosal damage induced by 5-FU chemotherapy in cancer-bearing mice, which may be related to its function in relieving oxidative stress injury and inhibiting apoptosis of colonic tissue.
目的: 观察电针“足三里”对大肠癌荷瘤小鼠5-氟尿嘧啶（5-FU）化疗后肠黏膜损伤的减轻作用及对结肠组织氧化应激及细胞凋亡的影响，探讨电针缓解化疗后肠黏膜损伤的部分机制。方法: BALB/c小鼠随机分为正常组、CT26组、5-FU组、非穴组和足三里组，每组6只。除正常组外，其余4组采用大肠癌CT26细胞皮下注射种植移植瘤；5-FU组、非穴组和足三里组腹腔注射 5-FU（5 mg/mL）溶液，每3 d注射 1次，共7次。足三里组和非穴组小鼠在每次给予5-FU腹腔注射后进行电针，足三里组电针双侧“足三里”，非穴组电针双侧非穴，每次5 min，3 d治疗1次，共治疗21 d。治疗结束后评估小鼠腹泻指数，测量小鼠结肠长度；HE染色法观察小鼠结肠黏膜病理形态，测量结肠绒毛长度；生化分析法检测小鼠结肠组织丙二醛（MDA）含量、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性；TUNEL法检测小鼠结肠组织细胞凋亡水平；免疫组织化学法检测小鼠结肠组织B淋巴细胞瘤-2蛋白（Bcl-2）、B淋巴细胞瘤-2基因相关X蛋白（Bax）阳性表达水平。结果: 与正常组比较，CT26组小鼠腹泻指数、结肠长度、结肠绒毛长度、MDA含量、SOD活性、GSH-Px活性、细胞凋亡率、Bax、Bcl-2阳性表达率差异均无统计学意义。与CT26组比较，5-FU组小鼠腹泻指数升高（P<0.01），结肠长度及结肠绒毛均缩短（P<0.01），结肠组织MDA含量升高（P<0.05），SOD和GSH-Px活性降低（P<0.01），细胞凋亡率升高（P<0.01），Bax阳性表达升高（P<0.05），Bcl-2阳性表达率降低（P<0.01）。与5-FU组比较，足三里组小鼠腹泻指数降低（P<0.01），结肠长度及结肠绒毛均增长（P<0.01），结肠组织MDA含量降低（P<0.05），SOD和GSH-Px活性升高（P<0.01），细胞凋亡率降低（P<0.01），Bax表达降低（P<0.05），Bcl-2表达升高（P<0.01）；非穴组小鼠结肠绒毛长度增长（P<0.01），结肠组织SOD、GSH-Px活性升高（P<0.05，P<0.01），结肠组织细胞凋亡率降低（P<0.01）。结论: 电针“足三里”可减轻大肠癌荷瘤小鼠5-FU化疗后肠黏膜损伤，其作用机制可能与调节结肠组织氧化应激、抑制细胞凋亡有关。.

Gastrointestinal dysfunction is manifested as digestive symptoms. Clinically, Zusanli (ST36) is crucial in the acupoint prescriptions of acupuncture no matter which type of the disease is differentiated in traditional Chinese medicine, but the underlying mechanism remains to be explored. Aiming to summarize the current status of the researches in terms of ameliorating gastrointestinal mucosal damage and regulating gastrointestinal motility disorders, we systematically reviewed the basic researches on the intervention with electroacupuncture (EA) at \"ST36\" in treatment of the diseases related to gastrointestinal dysfunction in the past 5 years, after searching the articles from Chinese and English databases. The results suggest that EA at ST36 may regulate the local gastrointestinal inflammation, oxidative stress and immune microenvironment to relieve gastrointestinal mucosal damage and adjust gastrointestinal motility disorders by means of modulating the central and peripheral nerve signaling as well as the function of mast cells and Cajal interstitial cells.
胃肠功能异常以消化道症状为主，无论何种类型，针灸临床中均选用足三里为重要腧穴处方，但其背后的科学机制仍有待探索。本文通过系统梳理近5年中、英文数据库中使用电针干预单穴“足三里”治疗胃肠功能异常相关疾病的基础研究，旨在从改善胃肠黏膜损伤和调节胃肠动力紊乱两个方面总结其研究现状。电针“足三里”可能以巨噬细胞为中心环节调节胃肠局部炎性反应、氧化应激和免疫微环境来改善胃肠黏膜损伤，并通过调节中枢和外周信号转导及肥大细胞和Cajal间质细胞功能来调节胃肠动力紊乱。.

OBJECTIVE: To observe the similarities and differences of effects of moxibustion at \"Zusanli\" (ST36) on target tissues and macrophages polarization in knee osteoarthritis (KOA) and rheumatoid arthritis (RA) rats, and to summarize its efficacy and characteristics.
METHODS: Thirty rats were equally and randomly divided into control, KOA, RA, KOA+Moxi and RA+Moxi groups. The KOA model and RA model were induced by injection of sodium monoiodoacetate or Freund\'s complete adjuvant into the rats\' knee joints, respectively. Rats of the KOA+Moxi and RA+Moxi groups received moxibustion stimulation at bilateral ST36 for 30 min, once a day for 21 days, beginning from the 7th day on after modeling. The contents of serum interleukin(IL)-1β and IL-10 were detected by ELISA. Histopathological changes (Markin score of the knee cartilage and synovial pathology score) of the knee joints were observed after HE staining. The polarization state of M1 and M2 macrophages in the synovial tissue of the knee joints was assessed by detecting the expression of CD86 and CD206 after immunofluorescence staining.
RESULTS: Compared with the control group, the content of serum IL-1β, synovial pathology score, and synovial CD86 expression were significantly increased (P<0.01, P<0.05), while the content of serum IL-10 and synovial CD206 expression markedly decreased (P<0.01) in both KOA and RA groups；the Markin score was increased (P<0.01) in the KOA group. In comparison with the KOA group, the Markin score was obviously decreased (P<0.01), while the content of serum IL-10 and CD206 expression were apparently increased (P<0.01) in the KOA+Moxi group. No significant changes were found in the content of serum IL-1β, synovial pathology score and CD86 expression in the KOA+Moxi group relevant to the KOA group. In comparison with the RA group, the content of serum IL-1β, synovial pathology score, and CD86 expression were considerably decreased (P<0.01) in the RA+Moxi group. No marked differences were found in the serum IL-10 level, Markin score, and CD206 expression between RA+Moxi and RA model groups. The increased Markin score was significantly higher in the KOA group than in the RA group (P<0.01), but the increased synovial pathology score was significantly lower in the KOA group than in the RA group (P<0.01). Correspondingly, the effect of moxibustion at ST36 was significantly better in RA model than in KOA model in reducing serum IL-1β (P<0.05).
CONCLUSIONS: Moxibustion at ST36 can effectively reduce cartilage injury of knee joint in rats with KOA and reduce synovial injury in rats with RA, which may be related with its effects in lowering IL-1β level in RA model by inhibiting the polarization of M1 macrophages, and up-regulating level of IL-10 in KOA model by promoting the polarization of M2 macrophages. However, the relevant mechanism needs to be further studied.
目的: 通过艾灸膝关节骨关节炎（KOA）与类风湿性关节炎（RA）大鼠“足三里”，观察膝关节滑膜巨噬细胞极化情况的异同，并归纳腧穴功效特点。方法: 雄性SD大鼠随机分为对照组、KOA组、RA组、KOA+艾灸组、RA+艾灸组，每组6只。采用膝关节注射单碘乙酸钠建立KOA模型，膝关节注射弗氏完全佐剂建立RA模型。造模后7 d给予KOA+艾灸组与RA+艾灸组大鼠艾灸“足三里”治疗，30 min/次，1 次/d，连续21 d。治疗结束后，采用ELISA法检测大鼠血清白细胞介素（IL）-1β、IL-10含量；HE染色法观察大鼠膝关节组织形态学变化，分别采用Markin评分和膝关节滑膜病理评分评价大鼠膝关节软骨和滑膜病理情况；免疫荧光染色法检测大鼠膝关节滑膜CD86、CD206表达，分析M1、M2巨噬细胞极化情况。结果: 与对照组相比，KOA组及RA组大鼠血清IL-1β含量、膝关节滑膜病理评分、滑膜CD86表达水平显著升高（P<0.01，P<0.05），血清IL-10含量及滑膜CD206表达水平显著降低（P<0.01）；KOA组膝关节Markin评分显著升高（P<0.01）。与KOA组比较，KOA+艾灸组大鼠血清IL-10含量和滑膜CD206表达显著升高（P<0.01），Markin评分显著降低（P<0.01）。与RA组比较，RA+艾灸组大鼠血清IL-1β含量、滑膜病理评分和CD86表达显著降低（P<0.01）。KOA组膝关节Markin评分显著高于RA组（P<0.01），滑膜病理评分和CD86表达显著低于RA组（P<0.01）。KOA+艾灸组血清IL-1β含量、膝关节Markin评分显著高于RA+艾灸组（P<0.05，P<0.01），滑膜病理评分显著低于RA+艾灸组（P<0.01）。结论: 艾灸“足三里”能有效减轻KOA大鼠与RA大鼠膝关节炎性损伤，调控炎性反应，但对KOA来说主要通过促进M2型巨噬细胞极化以提高抑炎因子IL-10含量，而对RA则是抑制M1型巨噬细胞极化从而降低促炎因子IL-1β，这可能与“足三里”能随着不同机体状态对相应靶器官进行靶向性调节有关。.

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) at \"Zusanli\" (ST36) on improving glucose metabolism disorder in chronic restraint induced depressed rats.
METHODS: A total of 30 male SD rats were randomly divided into control, model and EA groups, with 10 rats in each group. The depression model was established by chronic restraint 2.5 h each day for 4 weeks. For rats in the EA group, EA stimulation (1 mA, 2 Hz, 30 min) was applied to bilateral ST36 during the modeling period, once a day for 4 weeks. The body weight of the rats was recorded before and after modeling. The behavior of rats was observed by sugar-water preference and forced swimming after modeling. The contents of glucose and glycosylated albumin in serum were determined by biochemical method. The histopathological morphology and liver glycogen content were observed by HE and PAS staining. The expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated (p)-PI3K (p-PI3K), protein kinase B (Akt), p-Akt, glycogen synthase kinase-3β (GSK3β) and p-GSK3β proteins in liver were determined by Western blot.
RESULTS: Compared with the control group, the weight increment and sugar-water preference index decreased (P<0.01), the immobile swimming time was prolonged (P<0.01), the glucose and glycosylated albumin contents in serum increased (P<0.05), the expression of p-Akt protein and the ratio of p-Akt/Akt in liver tissues decreased (P<0.001), the expression of p-GSK3β protein and the ratio of p-GSK3β/GSK3β in liver tissues increased (P<0.01,P<0.001) in the model group. Compared with the model group, the weight increment and sugar-water preference index increased (P<0.05), the immobile swimming time was shortened (P<0.05), the glucose and glycosylated albumin contents in serum decreased (P<0.05), the expressions of p-PI3K and p-Akt proteins and the ratio of p-PI3K/PI3K and p-Akt/Akt in liver tissues increased (P<0.05), the expression of p-GSK3β protein and the ratio of p-GSK3β/GSK3β in liver tissues decreased (P<0.01) in the EA group. HE staining showed that the structure of the hepatic lobule was intact, no obvious inflammatory cell infiltration or fibrosis was observed in the lobule and interstitium, and no abnormalities were observed in the small bile duct, portal vein and artery in the portal area. PAS staining showed that the intensity of staining from the center of the hepatic lobule to the periphery of the hepatic lobule was gradually enhanced in the blank group, that is, the glycogen-rich granules in the hepatic cells were gradually increased; most of the hepatocytes were light colored and glycogen was lost significantly in the model group; while the intensity of hepatocyte staining increased, the staining intensity of the perilobular zone was weaker than that in the blank group, and the glycogen particles partially recovered in the EA group.
CONCLUSIONS: EA intervention can regulate glucose metabolism disorder in chronic restraint induced depressed rats through PI3K/Akt/GSK3β signaling pathway.
目的：探讨电针“足三里”改善应激后抑郁大鼠糖代谢紊乱的作用机制。方法：SD大鼠随机分为空白组、模型组、电针组，每组10只。采用慢性束缚应激复制抑郁大鼠模型。电针组造模期间同时进行双侧“足三里”电针干预，每次30 min，每日1次，持续4周。造模前后记录大鼠体质量并计算体质量增加量；造模结束后行糖水偏好实验、强迫游泳实验观察大鼠行为学；生化法检测大鼠血清中葡萄糖含量及糖化白蛋白水平；HE染色、PAS染色观察大鼠肝脏组织病理形态及肝糖原含量；Western blot法检测肝脏组织中磷脂酰肌醇3-激酶(PI3K)、磷酸化(p)-PI3K、蛋白激酶B(Akt)、p-Akt、糖原合成酶激酶-3(GSK3β)、p-GSK3β蛋白表达量。结果：与空白组相比，模型组大鼠体质量增加量、糖水偏好指数降低(P<0.01)，游泳不动时间延长(P<0.01)，血清中葡萄糖含量及糖化白蛋白水平升高(P<0.05)，肝脏组织中p-Akt蛋白表达量及p-Akt/Akt比值降低(P<0.001)，p-GSK3β蛋白表达量及p-GSK3β/GSK3β比值升高(P<0.01，P<0.001)；与模型组相比，电针组大鼠体质量增加量、糖水偏好指数明显升高(P<0.05)，游泳不动时间缩短(P<0.05)，血清中葡萄糖含量及糖化白蛋白水平降低(P<0.05)，肝脏组织中p-PI3K、p-Akt蛋白表达量及p-PI3K/PI3K、p-Akt/Akt比值升高(P<0.05)，p-GSK3β蛋白表达量及p-GSK3β/GSK3β比值降低(P<0.01)。HE染色显示，各组大鼠肝组织小叶结构完整，小叶和间质内未见明显炎性细胞浸润或纤维化，汇管区内小胆管、门脉及动脉未见异常。PAS染色显示，空白组大鼠肝组织自肝小叶中心(即小叶3带)至小叶周围带染色程度渐增强，即肝细胞富含糖原颗粒渐增多；模型组肝细胞大部分着色浅淡，糖原脱失显著；电针组肝细胞着色增加，肝小叶周围带染色强度较空白组弱，糖原颗粒部分恢复。结论：电针“足三里”可以通过PI3K/Akt/GSK3β信号通路调节慢性束缚致抑郁大鼠的糖代谢紊乱。.

Peptic ulcer disease (PUD) is a common disease and frequently encountered in the clinic. Accumulating evidence suggests that PUD is associated with the gastrointestinal microbiota. Electroacupuncture (EA) is an improved version of acupuncture, which can improve the clinical effect by increasing the stimulation and delivering appropriate electrical pulses to needles. This method has been widely used in the treatment of peptic ulcer disease. However, its effect on gastrointestinal microbiota remains unclear. Therefore, in the present study, the ameliorative effect of EA was evaluated on the gastroduodenal mucosa, and the regulatory effect of the gastroduodenal microbiota was assessed in PUD mice. A total of 48 male Kun Ming mice were randomly divided into the following groups: normal control group (NC), PUD model group (PUD), Shousanli group (LI10), and Zusanli group (ST36) (n=12). The mice in groups LI10 and ST36 were treated with EA at LI10 and ST36, respectively. This intervention was continued for 7 days. Subsequently, we evaluated the morphological changes in the gastric and duodenal mucosa, and specific indices were measured, including the contents of serum dopamine (DA), the trefoil factor (TFF), and the vasoactive intestinal peptide (VIP). In addition, the gastric and duodenal microbiota were assessed via 16S ribosomal DNA sequencing. The results indicated that EA at LI10 or ST36 significantly reduced the injury of the gastroduodenal mucosa in PUD mice. The gastric microbial community structure of the groups LI10 and ST36 was similar to that of the NC group following comparison with the microbial community structure of the PUD model group. Moreover, the abundance of Firmicutes in the stomach was decreased, whereas that of Bacteroidetes was increased, and the abundance of Firmicutes in the duodenum was decreased. Furthermore, the microbial diversity and richness of the gastric microbiota in group LI10 were also significantly increased, and the serum dopamine and trefoil factor levels in group ST36 were significantly increased. Therefore, it is suggested that EA ameliorating PUD is in association with improving the levels of DA and TFF and regulating the relative abundances of Firmicutes and Bacteroidetes in the gastric microbiota.

OBJECTIVE: To observe the effect of electroacupuncture (EA) at \"Zusanli\"(ST36) pretreatment on lung functions, inflammatory response, and levels of angiotensin-converting enzyme 2 (ACE2) and angiotensin (1-7) ［Ang (1-7)］ in rats with sepsis-induced acute lung injury (ALI), so as to explore its mechanisms underlying improvement of ALI.
METHODS: Thirty male SD rats were randomly divided into normal, model and EA groups (n=10 in each group). The sepsis-related ALI model was established by intraperitoneal injection of lipopolysaccharide (LPS, 5 mg/kg). Rats of the EA group received EA (4 Hz/20 Hz, 1-3 mA) stimulation at bilateral ST36 for 30 min, once each day, for 7 days before modeling. The lung functions including forced vital capacity (FVC), forced expiratory volume at 0.1 second (FEV0.1) and FEV0.3 were detected using a respiratory function detector for small animals at 3 h after modeling. The bronchoalveolar lavage fluid (BALF) was collected for assaying the contents of Ang (1-7), tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) using ELISA. The lung wet/dry weight (W/D) ratio, FEV0.1/FVC, and FEV0.3/FVC were calculated. The histopathological changes of lung tissues were displayed by hematoxylin-eosin (H.E.) staining. The expression of ACE2 and mitochondrial assembly receptor (MasR) mRNAs and proteins in the lung tissue was detected by fluorescence quantitative real-time PCR and Western blot, separately.
RESULTS: Following modeling, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and the expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly decreased (P<0.01), while the level of W/D ratio and TNF-α and IL-1β contents in the BALF significantly increased (P<0.01) in the model group relevant to the normal group. In comparison with the model group, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly increased (P<0.05, P<0.01), whereas the level of W/D ratio, and TNF-α and IL-1β contents in the BALF were significantly decreased (P<0.05, P<0.01) in the EA group. H.E. staining showed pulmonary interstitial edema and alveolar septum thickening with severe inflammatory cell infiltration in the model group, which was relatively milder in the EA group.
CONCLUSIONS: EA preconditioning at ST36 can improve pulmonary function in sepsis-related ALI rats, which may be related to its effects in inhibiting inflammatory response and up-regulating ACE2 and MasR expression and Ang (1-7) content in the lung tissue.
目的：观察电针“足三里”预处理对脂多糖(LPS)诱导的脓毒症急性肺损伤(ALI)大鼠炎性反应、血管紧张素转化酶2(ACE2)及血管紧张素(1-7)［Ang(1-7)］的影响, 探讨电针对脓毒症ALI的预防作用和可能机制。方法：将30只雄性SD大鼠随机分为正常组、模型组、电针组, 每组10只。采用腹腔注射LPS(5 mg/kg)复制脓毒症ALI大鼠模型。电针组予双侧“足三里”穴电针预处理, 每日1次, 每次30 min, 连续1周后造模。造模3 h后采用小动物呼吸功能检测仪检测大鼠肺功能；ELISA法检测支气管肺泡灌洗液(BALF)中Ang(1-7)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)的含量；计算肺湿/干重比(W/D)；HE染色法观察大鼠肺组织病理变化；Western blot法、实时荧光定量PCR法检测肺组织中ACE2、线粒体组装受体(MasR)蛋白和mRNA的表达。结果：与正常组比较, 模型组肺组织可见明显肺间质水肿、肺泡间隔增厚；用力肺活量(FVC)、第0.1秒用力呼气量(FEV0.1)、第0.3秒用力呼气量(FEV0.3)、FEV0.1占FVC之比(FEV0.1/FVC), FEV0.3占FVC之比(FEV0.3/FVC)均显著降低(P<0.01)；BALF内TNF-α、IL-1β含量显著升高(P<0.01), Ang(1-7)含量显著降低(P<0.01)；W/D显著升高(P<0.01)；肺组织内ACE2、MasR蛋白和mRNA表达水平均显著降低(P<0.01)。与模型组比较, 电针组肺间质水肿及肺泡间隔厚度均有改善；FVC、FEV0.1、FEV0.3、FEV0.1/FVC及FEV0.3/FVC均显著升高(P<0.05, P<0.01)；BALF内TNF-α、IL-1β含量显著降低(P<0.01), Ang(1-7)含量显著升高(P<0.01)；W/D显著降低(P<0.05)；肺组织内ACE2、MasR蛋白和mRNA表达水平均显著升高(P<0.05, P<0.01)。结论：电针“足三里”穴预处理对LPS诱导的脓毒症ALI大鼠具有抗炎、降低肺水肿和对肺功能保护的作用, 其机制可能与电针通过上调ACE2和MasR的表达及Ang (1-7)的含量从而抑制炎性反应有关。.

Background: Electroacupuncture is a common treatment for chronic atrophic gastritis (CAG) in China. We aimed to determine the effects of electroacupuncture at zusanli (ST36) on intestinal microbiota in CAG rats. Methods: In total, 42 SD rats were randomly divided into normal (NC, 10 rats) and model (MG, 32 rats) groups. Rats in the MG group were established as CAG disease models. After that, the rats in the MG group were randomly divided into CAG (10 rats), electroacupuncture (EA, 10 rats), and Vitacoenzyme (Vit, 10 rats) groups. Rats in the NC and CAG groups were subjected to a 30-min/d confinement for 4 weeks. Rats in the EA group were given electroacupuncture at zusanli for 30 min/d for 4 weeks. Rats in the Vit group were given Vitacoenzyme solution 10 ml/(kg d) for 4 weeks. Histopathological changes in the gastric mucosa were observed with hematoxylin and eosin staining, and the gene expression level of p53, Bcl-2, and c-myc was determined using the qPCR method. The 16S rDNA sequencing technique was used to determine structural changes and relative abundance expression of intestinal flora. Results: Compared with the NC group, gastric mucosal pathology in the CAG group revealed significant inflammatory infiltration, and the gastric mucosal lesions in the electroacupuncture group were improved remarkably; the expression of p53 and c-myc genes in the CAG group increased (p < 0.05), while the expression of Bcl-2 genes decreased (p < 0.05) in the EA group, that of p53 and c-myc genes decreased (p < 0.05), and that of Bcl-2 genes increased (p < 0.05). The abundance of bacteria such as Lactobacillus, Desulfobacterota, and Bacteroides pectinophilus group in the CAG group increased (p < 0.05), while that of bacteria such as Gastranaerophilales, Romboutsia, and Blautia decreased (p < 0.05). The relative abundance of Desulfobacterota and Helicobacter in the EA group decreased (p < 0.05), while that of probiotic bacteria such as Oscillospirales, Romboutsia, and Christensenellaceae increased (p < 0.05). Conclusion: Electroacupuncture at zusanli can promote the repair of pathological damage to the gastric mucosa in rats with CAG, and the mechanism might relate to the reduction in the relative abundance of harmful bacteria, increase in the relative abundance of intestinal probiotics, and regulation of the intestinal microbiota.

OBJECTIVE: To observe the effect and safety of injection of vitamin B1 into Zusanli (ST36) and Hegu (LI4) in the treatment of functional dyspepsia (FD).
METHODS: A total of 100 FD patients were equally divided into medication group and acupoint injection group (n=50/group) according to a random number table. Patients in the medication group were ordered to take mosapride citrate tablets (5 mg) orally 30 min before each of the three meals, while those in the acupoint injection group received injection of vitamin B1 into ST36 and LI4, once every other day, three times a week. One week later, the clinical symptom scores, total effective rates, serum gastrin (GAS) and plasma motilin (MTL) contents, and gastric emptying rates between the two groups were compared, followed by the observation of adverse reactions. Two weeks\' follow-up survey was conducted after the end of treatment, and the clinical symptom scores and total effective rates of the two groups were further compared.
RESULTS: Compared with the data before treatment in the same one group, the clinical symptom scores and serum GAS contents of post-treatment as well as the follow-up symptom scores were all significantly decreased (P<0.05，P<0.01), while the plasma MTL levels and gastric emptying rates were obviously increased in both groups (P<0.01). Comparison between the two groups showed that the clinical symptom score, serum GAS content after the treatment and follow-up symptom score were considerably lower (P<0.01), but the total effective rate, plasma MTL, gastric emptying rate after the treatment and total effective rate of follow-up notably higher in the acupoint injection group than those in the medication group (P<0.01,P<0.05).
CONCLUSIONS: Injection of vitamin B1 into ST36 and LI4 is effective in improving symptoms of PD patients, which may be related to its functions in regulating the levels of GAS and MTL in blood, and facilitating gastrointestinal motility.
目的：观察足三里、合谷穴位注射维生素B1治疗功能性消化不良（FD）的疗效及安全性。方法：选择FD患者100例，按照随机数字表法分为西药组和穴位注射组，每组50例。西药组于每日三餐前30 min口服枸橼酸莫沙必利片5 mg，共1周；穴位注射组给予足三里、合谷穴位注射维生素B1，隔日1次，共3次。1周后比较两组临床症状积分、总有效率、血清胃泌素(GAS)含量、血浆胃动素(MTL)含量及胃排空率，观察两组患者的不良反应。结束治疗后2周随访，比较两组临床症状积分及总有效率。结果：与本组治疗前比较，两组治疗后、随访时临床症状积分、血清GAS含量均降低(P<0.05，P<0.01)，血浆MTL含量、胃排空率升高(P<0.01)。与西药组比较，穴位注射组治疗后、随访时临床症状积分、血清GAS含量降低(P<0.01)，血浆MTL含量、胃排空率升高(P<0.01)。穴位注射组治疗后、随访时总有效率分别为91.84%（45/49）、93.88%（46/49），西药组治疗后、随访时总有效率分别为75.51%（37/49）、77.55%（38/49），穴位注射组临床疗效优于西药组（P<0.05）。结论：足三里、合谷穴位注射维生素B1治疗FD疗效显著，机制可能与其调节胃肠道激素GAS、MTL含量，增强胃肠动力有关。.