Objective:The present study was designed to investigate whether transfecting DC with tumor derived total RNA is an effective way to induce CTL and antitumor immunity.Methods:DC were propagated from bone marrow(BM) of C57BL/6J(H 2k b?I A b)mice in vitro with GM CSF+IL 4.Tumor derived total RNA extracted from actively growing Hepa 1 6 cells was used to transfected DC.The phenotypes of DC were detected by FACS,the cytotoxicity of CTL was assayed by MTT method.Results:The tumor derived total RNA transfected DC exhibited much more and longer plasm membrane processes and increased expression of MHC Ⅰ?MHC Ⅱ?CD80(B7 1)?CD86(B7 2).Conclusion:This experiment has shown that DC transfected with tumor derived total RNA of C57BL/6J cells could stimulate effectively the responsiveness of syngenic splenic T cells to induce specific CTL against C57BL/6J cells.

OBJECTIVETo observe the effects of electroacupuncture (EA) at different acupoints on apoptosis-related serum and expression of microRNA (miRNA) in rats with myocardial ischemia, so as to explore its mechanism of action.

METHODSA total of 48 male Wistar rats were randomly divided into a normal group, a model group, a Neiguan group and a acupoint compatibility group, 12 rats in each group. Isoprenaline hydrochloride (ISO) with a daily dose of 2 mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group, Neiguan group and acupoint compatibility group. Rats in the normal group were subcutaneously injected with an equal volume of normal saline. After modeling, rats in the Neiguan group were treated with EA at "Neiguan" (PC 6), while rats in the acupoint compatibility group were treated with EA at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Neiguan" (PC 6). Rats in the normal group and model group were treated with immobilization, once day for 21 days. The contents of creatine kinase-MB (CK-MB), vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) in serum were detected by enzyme-linked immunosorbent assay (ELISA); apoptosis index (AI) of myocardial cells was detected by TUNEL method; the expressions of miRNA-1, miRNA-133, miRNA-208 and miRNA-499 were detected by real-time PCR method.

RESULTSCompared with the normal group, the serum CK-MB, VCAM-1 and ET-1 were significantly increased in the model group, Neiguan group and acupoint compatibility group (all P < 0.01), and the apoptosis index was significantly increased (all P < 0.01). The CK-MB, VCAM-1 and ET-1 in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group (all P < 0.01); the AI was reduced, which was more significant in the acupoint compatibility group (P < 0.05). Compared with the normal group, the expression of miRNA-133 was reduced (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly increased in the model group (all P < 0.01). Compared with the model group, the expression of miRNA-133 was increased (both P < 0.01) and that of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced (all P < 0.01) in the Neiguan group and acupoint compatibility group. Compared with the Neiguan group, the expression of miRNA-133 was increased (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced in the acupoint compatibility group (P < 0.01, P < 0.05).

CONCLUSIONEA at acupoints, especially acupoint compatibility group, could effectively prevent and treat myocardial ischemia, and the protective effect is possibly correlated to the double regulation on increasing the expression of miRNA-133 and inhibiting the expression of miRNA-1, miRNA-208, miRNA-499.

Acupuncture Points ; Animals ; Apoptosis ; Disease Models, Animal ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Myocardial Ischemia ; genetics ; metabolism ; physiopathology ; therapy ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

Objective The present study was designed to investigate whether pulsing DCs with tumor-derived extracts is an ef- fective way to induce CTL. and antitumor immunity, Methods DCs were propagated from bone marrow (BM)of C57BL/6J(H-2Kb. I- Ab)mice in vitro with GM-CSF + IL-4tumor associated antigen (TAA) extracted from actively growing Hepa 1-6 cells was used to activate DCs. The phenotypes of DCs were detected by FACS, the cytotoxicity of CTL was as- sayed by 3H-TdR labbel assay. Result and Conclusion The TAA extract pulsed DCs exhibited much more and longer cell processes and increased expression of MHC- Ⅰ, MHC-Ⅱ, CD80 (B7-1 ) 、 CD86 (B7-2 ). This experiment has shown that DCs pulsed with TAA extracts of C57B/6J cells could stimulate effectively the responsiveness of syngenic splenic T cells to induce specific CTL against C57BL/6J cells.

Objective: To investigate the mechanism of miR-503 modulates radio-resistance of esophageal squamous cell carcinoma (ESCC) by targeting excision-repair cross-complementing 1 (ERCC1). Methods: The expression level of miR-503 in radio-resistant ESCC tumor tissues and KYSE140 and KYSE140R cells was detected by qPCR. The miR-503 mimic, miR-503 inhibitor or si-ERCC1 was transfected into KYSE140 and KYSE140R cells.After radiation treatment, the colony formation assay and CCK-8 assay were used to detect the proliferation of KYSE140R cells. Flow cytometry was used to detect apoptosis of KYSE140R cells. WB was used to detect changes in protein expression of ERCC1. Dual luciferase reporter gene assay was used to validate the targeting relationship between miR-503 and ERCC1. Results: The expression level of miR-503 was down-regulated in radio-resistant tissues and ESCC cell lines (all P<0.01). Over-expression of miR-503 significantly inhibited cell proliferation and promoted apoptosis of KYSE140R cells (all P<0.01). Dual-luciferase reporter assay validated that ERCC1 was a target gene of miR-503, and miR-503 negatively regulated the expression of ERCC1. Over-expression of miR-503 significantly down-regulated the expression of ERCC1 in KYSE140 and KYSE140R cells (both P<0.01), inhibited cell proliferation (both P<0.01), but significantly increased apoptosis rate (all P<0.01); knockdown of ERCC1 exhibited a similar effect, while knockdown of both ERCC1 and miR-503 reversed the above effects. Conclusion: Over-expression of miR-503 up-regulated the radio-sensitivity of KYSE140R cells by targeting ERCC1.

OBJECTIVETo explore the effects of principal-subordinate acupoints combination on improving myocardial ischemia, and the gene regulatory pathways for the protection of myocardial ischemia.

METHODSAccording to the random number table method, 70 SPF Wistar male rats were divided into a normal group, a model group, a LY294002 group, an insulin-like growth factors-1(IGF-1) group, a Neiguan group, an acupoint combination group and an acupoint combination + LY294002 group, 10 rats in each one. Rats in the normal group were injected with 0.9% NaCl solution, while rats in the remaining groups were treated with abdominal subcutaneous injection of isoroterenol hydrochloride to establish the rat model of myocardial ischemia. Rats in the LY294002 group and IGF-1 group were treated with injection of LY294002 solution and IGF-1 solution for 14 days. Rats in the Neiguan group were treated with electroacupuncture (EA) at "Neiguan" (PC 6) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination group were treated with EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination + LY294002 group were treated with LY294002 solution for 14 days, and EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) was given before model establishment, once a day for 21 days. EA pretreatment was given before model establishment in all acupuncture groups. The heart rate (HR) and ST segment voltage were detected before and after treatment; the myocardial pathological morphology was observed by HE staining; the expressions of P13K mRNA and Akt mRNA were tested.

RESULTSAfter modeling, HR and ST segment voltage in all intervention groups were higher than those in the normal group (all P < 0.01); after the intervention, the HR and the ST segment voltage in the acupoint combination group, IGF-1 group and IGF-1 group were improved (P < 0.01, P < 0.05), which was more significant in the acupoint combination group and Neiguan group (all P < 0.01). As for the myocardial pathological morphology, obvious myocardial ischemia was observed in the model group, and that in the LY294002 group was the most serious, and that in the acupoint combination+ LY294002 group was moderate. After intervention, the myocardial pathological damage in the IGF-1 group, Neiguan group and acupoint combination group was significant improved, which was more significant in the IGF-1 group and acupoint combination group. As for the expression of PI3K mRNA and Akt mRNA, compared with normal group, the expression of PI3K mRNA was increased in the remaining groups after modeling (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0. 01). The expression of Akt mRNA in the LY294002 group and acupoint combination + LY294002 group was reduced (P < 0. 01, P < 0.05), while that in the remaining groups was increased (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0.01).

CONCLUSIONThe principal-subordinate acupoints combination could improve heart rate and ST segment voltage in rats with chronic myocardial ischemia, reduce myocardial pathological damage, which is superior to single selection of "Neiguan" (PC 6). The PI3K/Akt signaling pathway may be involved in the regulation mechanism of principal-subordinate acupoints combination for the protection of chronic myocardial ischemia.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Chronic Disease ; therapy ; Electroacupuncture ; Electrocardiography ; Heart Rate ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Myocardial Ischemia ; enzymology ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rats ; Rats, Wistar

Continuous cropping obstacle is the bottleneck of medicinal plant cultivation, which seriously affects the quality and yield of medicinal materials. The research on the mechanism of continuous cropping obstacle has evolved from soil physical and chemical properties and allelopathy in the 1970s to the changes of rhizosphere microenvironment and plant response mechanism at present. According to the available studies in this field and our previous research work, we systematically analyzed the mechanism of rhizosphere exudate-mediated microbial community reconstruction in the soil of the medicinal plants in continuous cropping. Specifically, rhizosphere exudates, providing the carbon source and energy for microbial growth, act as inducers or repellents to induce microbial growth or transfer, thereby changing the physicochemical properties (such as acidity) of rhizosphere soil and further altering the structure of rhizosphere microbial community. Further, we comprehensively discussed the ways of synergism between rhizosphere exudates and soil microorganisms in causing harm to the medicinal plants in continuous cropping. That is, rhizosphere exudates mediate the infection of the rhizosphere by pathogenic microorganisms, increase the susceptibility of the nearby plants, inhibit the defense of the host plants, and protect the pathogens to occupy the dominant niche. The synergistic interaction results in the release of more pathogenic factors such as mycotoxins by rhizosphere pathogens, enhanced toxicity of rhizosphere allelochemicals, and deterioration of soil physical and chemical properties. This paper summarizes the role of interaction between rhizosphere exudates and soil microorganisms in the formation of continuous cropping obstacles, aiming to provide a new research idea for revealing the formation mechanism as well as the theoretical support for overcoming continuous cropping obstacles of medicinal plants.