Search

Objectives: To observe the protective effect of acupuncture on ovarian function and its impact on key molecules (Yes associated protein ［YAP］ and transcriptional coactivator with PDZ binding motif ［TAZ］) in the Hippo signaling pathway in poor ovarian response (POR) mice, in order to explore its mechanisms underlying improvement of POR by inhibiting ovarian cell apoptosis., Methods: The mice with regular motility cycle after screening were randomly divided into normal control, model and acupuncture groups, with 12 mice in each group. The POR model was established by gavage of tripterygium wilfordii polyglycoside suspension (50 mg · kg -1 ·d -1 ) for 2 weeks, and the mice of the normal control group was received an equal volume of 0.9% sodium chloride solution by gavage. The mice in the acupuncture group received manual acupuncture stimulation of bilateral "Taichong" (LR3) and "Sanyinjiao" (SP6), and "Zhongji" (CV3) and "Guanyuan" (CV4), with the filiform needles retained for 20 min, once daily for successive 10 days. The estrous cycle was determined by using vaginal exfoliated cell smear, and the body mass was detected weekly. The levels of serum anti Mullerian hormone (AMH), follicle stimulating hormone (FSH), estradiol (E2) and luteinizing hormone (LH) were measured using enzyme-linked immunosorbent assay (ELISA). Histopathological changes of the ovarian tissue were observed after H.E. staining, and the apoptosis of ovarian granulosa cells was measured using terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. The expression levels of YAP, TAZ, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), cysteine aspartic acid specific protease-3 (Caspase-3) mRNAs and proteins were detected using real-time PCR and Western blot, respectively., Results: Compared with the normal control group, the model group had an increase in the rate of estrous cycle disorder, estrous cycle, serum FSH and LH content, and apoptosis of ovarian granulosa cells, and expression levels of Bax and Caspase-3 mRNAs and p-YAP, Bax and Caspase-3 proteins ( P <0.01), and a decrease in the body mass, number of retrieved oocytes, ovarian wet weight and ovarian index, serum AMH and E2 contents, and the expression levels of YAP, TAZ, Bcl-2 mRNAs and proteins ( P <0.01). After acupuncture intervention, modeling induced increase and decrease of indexes mentioned above were completely reversed ( P <0.05, P <0.01). H.E. staining showed deformed ovarian structure, reduced number of normal developing follicles and increased number of atretic follicles, disordered arrangement of the granulosa cells with fewer hierarchy in the model group, which was improved in the acupuncture group, such as increase in the number and improvement in the shape of normal ovarian follicles and reduction of the atretic follicles., Conclusions: Acupuncture intervention can inhibit the apoptosis of ovarian granulosa cells and improve the ovarian function of POR mice, which may be related to its effects in up-regulating the expressions of YAP, TAZ (key molecules of Hippo signaling pathway).

Objectives: To observe the effect of moxibustion with seed-size moxa cones on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factor-κB(NF-κB) signaling pathway in mice with ulcerative colitis(UC), so as to explore the therapeutic mechanism of moxibustion with seed-size moxa cones on colonic injury in UC., Methods: Forty male C57BL/6 mice were randomly divided into blank group, model group, moxibustion group, and western medicine group, with 10 mice in each group. The UC mouse model was established by 3% DSS solution by free drinking for 7 consecutive days. Mice in the moxibustion group were treated with seed-size moxa cones at "Zhongwan"(CV12), "Tianshu"(ST25) and "Shangjuxu"(ST37), 3 moxa cones per point, with each cone applied for approximately 30 s, while mice in the western medicine group were orally administered with 300 mg/kg mesalazine solution, which were both conducted once a day for 7 consecutive days. The general condition of mice was observed every 2 days, and the disease activity index (DAI) score was calculated. HE staining was used to observe the morphology of colonic tissue in mice. ELISA was used to detect the serum interleukin(IL)-1β, tumor necrosis factor(TNF)-α, IL-6, and IL-8 contents. Immunohistochemistry was used to detect the positive expression of TLR4 and MyD88 in colonic tissue of mice. Real-time fluorescence quantitative PCR was used to detect the expression levels of TLR4, MyD88, and NF-κB p65 mRNAs in colonic tissue., Results: Compared with the blank group, varying degrees of soft or watery stools were observed, colon length and body weight were decreased( P <0.01) in mice of the model group, while DAI score, colon weight index, mucosal damage score, colonic pathological score, serum IL-1β, TNF-α, IL-6, and IL-8 contents, positive expressions of TLR4 and MyD88, and TLR4, MyD88, and NF-κB p65 mRNA expressions in colonic tissue were increased( P <0.01). Compared with the model group, improved fecal characteristics were observed, colon length and body weight were increased( P <0.01) in mice of the moxibustion group and western medicine group, while DAI scores, colon weight indexes, mucosal damage scores, colonic pathological score, serum contents of IL-1β, TNF-α, IL-6, and IL-8, positive expressions of TLR4 and MyD88, and TLR4, MyD88, and NF-κB p65 mRNA expressions in colonic tissue were decreased( P <0.01, P <0.05). There was no significant difference in the above indicators between the moxibustion group and the western medicine group., Conclusions: Moxibustion with seed-size moxa cones may alleviate colonic injury in UC mice by regulating the TLR4/MyD88/NF-κB signaling pathway and reducing the release of inflammatory factors.

Objectives: To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC., Methods: Twenty-six male SD rats were randomly divided into 4 groups：normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1β, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions., Results: Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions( P <0.01, P <0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1β, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions( P <0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group( P <0.01, P <0.05)., Conclusions: EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.

Objective: To observe the anti-tumor effect of moxibustion with seed-sized moxa cones on Hepa1-6 liver cancer bearing (HLCB) mice and its regulatory mechanism on cell apoptosis., Methods: A total of 40 male C57BL/6 mice were randomly divided into control, moxibustion, cyclophosphamide (CTX) and moxibustion+CTX groups, with 10 mice in each group. The HLCB model was established by subcutaneous inoculation of Hepa1-6 cancer cells into the right armpit. Mice of the CTX and moxibustion+CTX groups were given intraperitoneal injection of CTX (30 mg/kg), once daily for 3 days. Moxibustion with seed-sized moxa cones were applied to "Dazhui" (GV14), bilateral"Zusanli" (ST36) and "Sanyinjiao" (SP6), with 5 moxa cones for each acupoint, once daily for 10 consecutive days. The survival status scores and body weight of HLCB mice were observed, and the tumor weight and tumor inhibition rate were detected. HE staining was used to observe the morphological changes of tumor tissue. ELISA was used to detect the levels of serum interleukin (IL)-2, IL-4 and tumor necrosis factor-α (TNF-α). Western blot and fluorescent quantitative real-time PCR were used to detect the protein and mRNA expressions of cysteine aspartate protease (Caspase) -3 and Caspase-9 in tumor tissues, separately., Results: Compared with the control group, the survival status scores, body weight, serum IL-2 and TNF-α levels were significantly increased ( P< 0.05, P< 0.01), the tumor weight and serum IL-4 levels were significantly decreased ( P< 0.05) in the moxibustion group；while the survival status, body weight, tumor weight, serum IL-2 and IL-4 levels were significantly decreased ( P< 0.01, P< 0.05), the content of TNF-α was significantly increased ( P< 0.01) in the CTX group. The protein and mRNA expressions of Caspase-3 and Caspase-9 in the 3 trentment groups were significantly increased ( P< 0.05, P< 0.01). In comparison with the moxibustion group, the survival status scores, body weight and tumor weight, serum content of IL-2 were significantly decreased ( P< 0.01, P< 0.05). In contrast to the CTX group, the survival status scores, body weight, serum IL-2 and TNF-α content, and the expressions of Caspase-3 and Caspase-9 in tumor tissue were significantly increased ( P< 0.01, P< 0.05), and the tumor weight and serum IL-4 content were significantly decreased ( P< 0.05) in the moxibustion+CTX group. Results of HE staining showed that the tumor cells in the control group had clear nuclear membranes and nucleoli, with more dividing cells； while less nuclear division and an increase in tumor necrosis areas were found in the 3 treatment groups., Conclusion: Moxibustion with seed-size moxa cone can enhance the anti-tumor effect of CTX and improve the quality of life of HLCB mice, which may be related with its effect in activating the expressions of Caspase-3 and Caspase-9 in tumor tissue.

Objective: To observe the effect of electroacupuncture （EA） at "Zhongwan" （CV12）, "Tianshu" （ST25） and "Shangjuxu" （ST37） （an acupoint prescription "Changbingfang" for treatment of intestinal disorders） on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis （UC）, so as to explore its mechanism underlying improvement of UC., Methods: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation （10 Hz/50 Hz） was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension （200 mg/kg） once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index （DAI） score （0-4 points）. Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 （IL-6）, tumor necrosis factor- α （TNF- α ） and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues., Results: Compared with the control group, the DAI score, contents of serum IL-6 and TNF- α , the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased （ P <0.01）； while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were decreased （ P <0.01, P <0.05） in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF- α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were reversed in both medication and EA groups （ P <0.01, P <0.05）. The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK, p62 mRNA expression （ P <0.01, P <0.05）, and in down-regulating ratio of p-mTOR/mTOR （ P <0.05）. H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups., Conclusion: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.

Objective: To observe the effect of electroacupuncture (EA) of "Zhongwan"(CV 12), "Tianshu"(ST 20) and "Shangjuxu"(ST 37) (an acupoint prescription "Changbingfang" for treatment of intestinal disorders) on the expression of colonic nuclear factor kappaB p65 (NF-kappaB p65) protein expression and serum interleukin-4 (IL-4) content in rats, so as to explore its mechanism underlying improvement of ulcerative colitis (UC)., Methods: Thirty SD rats were randomly and equally divided into normal control, model and EA groups (10 rats/group). The UC model was made by enema with 8% acetic acid. EA stimulation (6 Hz/30 Hz) was delivered to "Zhongwan" (CV 12), "Tianshu" (ST 20) and "Shangjuxu" (ST 37) for 15 min separately, 0 h, 24 h and 48 h post-enema. The blood and colon tissues were taken for detection of serum IL-4 content with ELISA, and for assaying colonic NF-kappaB p65 protein expression with immunohistochemistry and Western blot, respectively., Results: Compared with the normal control group, colonic NF-kappaB p65 protein expression level shown by immunohistochemistry and Western blot and serum IL-4 content were significantly higher in the model group (P < 0.01, P < 0.05). After EA stimulation, colonic NF-kappaB p65 protein expression levels shown by both immunohistochemistry and Western blot were down-regulated significantly (P < 0.01, P < 0.05) and serum IL-4 content was up-regulated further in comparison with the model group (P < 0.01)., Conclusion: EA of acupoint recipe "Changbingfang" can down-regulate the proinflammatory NF-kappaB p65 protein expression and up-regulate serum anti-inflammatory cytokine IL-4 level in rats, which may contribute to its effect in improving ulcerative colitis.

Objective: To observe the effect of electroacupuncture (EA) stimulation of differently combined acupoints of "Zhongwan" (CV 12), "Tianshu" (ST 25) and "Shangjuxu" (ST 37, for treatment of enteral disorders) on acetic acid-induced intestinal mucosal lesion in rats so as to select the best combinations of acupoints., Methods: A total of 126 SD rats were divided into nine groups: normal control (normal), model, CV 12, ST 25, ST 37, CV 12 + ST 37, ST 25 + ST 37, CV 12 + ST 25 and CV 12 + ST 25 + ST 37. Intestinal mucosal lesion model was established by using enema (8% acetic acid 1 mL). EA was applied to the above-mentioned acupoints for 15 min, once daily for 3 days. General conditions, colonic mucosal damage index (CMDI) and pathological damage integral (PDI) were detected respectively. The colonic mucosal membrane tissue was stained with H. E. stain for observing its morphological changes under light microscope, and the ultrastructural changes were observed using electron transmission microscope., Results: Compared with the control group, CMDI was decreased significantly (P < 0.05) and PDI of the colonic mucosal membrane increased significantly in the model group (P < 0.05). Compared with the model group, CMDI was decreased markedly and PDI increased significantly (P < 0.05) in the CV 12, ST 25, ST 37, CV 12 + ST 37, ST 25 + ST 37, CV 12 + ST 25 and CV 12 + ST 25 + ST 37 groups (P < 0.05). Comparison among the 7 EA treatment groups showed that both CMDI and PDI of the CV 12 + ST 25 + ST 37 group were significantly superior to those of CV 12, ST 25, ST 37, CV 12 + ST 37, ST 25 + ST 37 and CV 12 + ST 25 groups (P < 0.05). Under light microscope, the colonic mucosal injury was remarkable, while those of the 7 EA treatment groups were relatively milder. Ultramicrostructural findings showed that the colonic mucosal lesion including shortening or sparseness of the intestinal epithelial villi, swelling of mitochondria with arrangement disorder, breaking or fusion of the cristae, and enlargement of the rough endocytoplasmic reticulum was found in the model group, while the situations were lighter in the 7 EA groups particularly in the CV 12 + ST 25 + ST 37 group., Conclusion: EA of CV 12, ST 25 and ST 37 acupoint alone or combination can relieve acetic acid-induced intestinal mucosal lesion in rats, and the effect of joint application of these 3 acupoints is obviously better, suggesting their clinical applicability in clinical treatment of intestinal disorders as a basic formula.

The characteristics and advantages of acupuncture therapy for apoplexy have been widely approved in China. However, the factors affecting the curative effect are various. More and more researchers think highly of the acupuncture needle-retaining duration. The present article makes a discussion on the significance of needle-retaining duration for improving stroke patients' symptoms and signs; and reviews the recent progresses, the extant problems and the academic significance about the studies of the abovementioned topic by combining the authors' previous work. Generally, a systemic study on this topic is rather deficient up to now, and is definitely necessary for further improving clinical therapeutic effect in the light of the requirements of evidence-based medicine.

Objective: To compare the effects of electroacupuncture (EA) stimulation of "Gastric Problem Formula" ["Neiguan" (PC 6), "Zhongwan" (CV 12) and "Zusanli" (ST 36)] on electrical activities of neurons of medullary subnucleus reticularis dorsalis (SRD) of the rat., Methods: Experiments were performed on SD rats anesthetized by 10% urethane (1.0-1.5 g/kg). Under artificial ventilation and immobilization (2% gallamine triethiodide, 5 ml/kg), extracelluar discharges were recorded by glass micropipettes inserted into SRD of the medulla (Caudal to the Obex: - 1.0 - -2.0 mm, L/R= 0.5-1.5 mm, H: 0.6- 1.5 mm below the medullary surface). Following getting stable recordings, the acupuncture needle inserted in "Neiguan" (PC 6) was manipulated moderately (twisting range: 120 degrees, frequency = 80 times/min) for 30 s. The same manipulation was performed at "Zhongwan" (CV 12) and "Zusanli" (ST 36) separately., Results: After moderate acupuncture stimulation, in the recorded 13 SRD neurons, the discharges were increased from (1.67 +/- 0.35) spikes/s to (17.55 +/- 1.40) spikes/s for "Neiguan" (PC 6), from (2.35 +/- 0.43) spikes/s to (16.96 +/- 1.83) spikes/s for "Zhongwan" (CV 12), and from (1.83 +/- 0.37) spikes/s to (17.39 +/- 1.58) spikes/s for "Zusanli" (ST 36). No apparent differences were found among 3 acupoints in their responses to manual manipulation., Conclusion: Moderate acupuncture stimulation of "Neiguan" (PC 6), "Zhongwan" (CV 12) and "Zusanli" (ST 36) can effectively activate spontaneous discharges of SRD neurons in the rat.

Objective: To select the best combination of the commonly-used acupoints [Zhongwan (CV 12), Neiguan (PC 6), and Zusanli (ST 36)] for treatment of gastric disorders., Methods: Forty-eight Wistar rats were randomly divided into 8 groups: model, ST36, CV12, PC6, ST36 + PC6, ST36 + CV12, CV12 + PC6 and ST36 + CV12 + PC6 by factorial design method. Acute gastric mucosal lesion model was established by intragastric perfusion of anhydrous alcohol. EA(1- 3 mA, 30 Hz) was applied to the above-mentioned acupoints for 20 min respectively. And after EA, gastric mucosal lesion index (LI), pathological lesion integral score (PLIS) were calculated and mucosal ultramicrostructural changes observed by optical microscope and transmission electron microscope respectively., Results: Compared with model group, the gastric mucosal LI and PLIS of ST36, CV12, PC6, ST36 + CV12, ST36 + PC6, CV12 + PC6, ST36 + CV12 + PC6 groups all decreased significantly (P < 0.05). Comparison among 7 EA groups showed that both LI and PLIS of ST36 + CV12 + PC6 group were significantly lower than those of ST36, V12, PC6, ST36 + CV12, ST36 + PC6 and CV12 + PC6 groups (P < 0.05). No significant differences were found between every two groups of ST36, CV12, PC6, ST36+ CV12, ST36 + PC6 and CV12 + PC6 groups (P > 0.05) in both LI and PLIS. Ultrastructural results showed that the injury of gastric mucosal parietal cells and delomorphous cells including mitochondrial swelling, crista arrange disorder and breaking, enlargement of the rough endoplasmic reticulum, ribosome exfoliation, etc. in model group was serious, but lightened significantly in EA groups particularly in ST36 + CV12 + PC6 group., Conclusion: ST36, PC6 and CV12 combination can effectively reduce anhydrous alcohol-induced gastric mucosal lesion in rats, and may be used as a basic formula

In this article, the authors make an analysis on the regularity of correlation between pathological reactions of acupoints and Zangfu organs/meridians, current clinical application and mechanism research, and summarize the relative specificity of the correlation between abnormal reactions of acupoints and states of Zangfu organs/meridians. In addition, the authors also point out some shortcomings in current researches on the above-mentioned aspects, for examples, only observation on the phenomenon without definite conclusion on the regularity of acupoint reactions, being restricted in the objective specificity, etc.. They stress that we should sum up a series of regularities for clinical application (early diagnosis), and conduct profound research on its underlying mechanism in the coming days.