[Moxibustion with seed-size moxa cone enhances anti-tumor effect of cyclophosphamide on Hepa1-6 hepatoma-bearing mice by regulating apoptosis factor cysteine family].

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.

[Effect of wheat-grain moxibustion on Wnt/ß-catenin signaling pathway in bone marrow cell in mice with bone marrow inhibition].

OBJECTIVE: To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) on Wnt/ß-catenin signaling pathway in bone marrow cell in mice with bone marrow inhibition, and to explore the possible mechanism of wheat-grain moxibustion in treating bone marrow inhibition. METHODS: Forty-five SPF male CD1(ICR) mice were randomly divided into a blank group, a model group and a wheat-grain moxibustion group, 15 mice in each group. The bone marrow inhibition model was established by intraperitoneal injection of 80 mg/kg of cyclophosphamide (CTX). The mice in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6), 3 moxa cones per acupoint, 30 s per moxa cone, once a day, for 7 consecutive days. The white blood cell count (WBC) was measured before modeling, before intervention and 3, 5 d and 7 d into intervention. After intervention, the general situation of mice was observed; the number of nucleated cells in bone marrow was detected; the serum levels of interleukin-3 (IL-3), interleukin-6 (IL-6) and granulocyte macrophage colony stimulating factor (GM-CSF) were measured by ELISA; the protein and mRNA expression of ß-catenin, cyclinD1 and C-Myc in bone marrow cells was measured by Western blot and real-time PCR method. RESULTS: Compared with the blank group, the mice in the model group showed sluggish reaction, unstable gait, decreased body weight, and the WBC, number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were decreased (P<0.01), and the protein and mRNA expression of ß-catenin, cyclinD1 and C-Myc was decreased (P<0.01). Compared with the model group, the mice in the wheat-grain moxibustion group showed better general condition, and WBC, the number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were increased (P<0.01, P<0.05), and the protein and mRNA expression of ß-catenin, cyclinD1 and C-Myc was increased (P<0.05). CONCLUSION: Wheat-grain moxibustion shows therapeutic effect on bone marrow inhibition, and its mechanism may be related to activating Wnt/ß-catenin signaling pathway in bone marrow cells, improving bone medullary hematopoiesis microenvironment and promoting bone marrow cell proliferation.

[Academic origin of round magnetic needle and standardization operation].

The origin and development of round magnetic needle was explored, and the structure of round magnetic needle was introduced in detail, including the handle, the body and the tip of the needle. The clinical opera tion of round magnetic needle were standardized from the aspects of the methods of holding needle, manipulation skill, tapping position, strength of manipulation, application scope and matters needing attention, which laid foundation for the popularization and application of round magnetic needle.

[Effects of electroacupuncture at acupoints "changbingfang" on serum IL-4 content and colonic NF-kappaB p65 expression in rats with ulcerative colitis].

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.

[Selection of basic acupoints combined with "enteral-disorder-formuia" for electroacupuncture treatment of intestinal mucosal lesion in rats].

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.