OBJECTIVE To investigate enhanced immune function of methionine encephalin (MENK) and its anti-tumor mechanism in CT26 colon cancer mouse model. METHODS 3×106 CT26 cells were implanted subcutaneously in BALB/c mice. Four days after, MENK was peritoneally administrated at the concentration of 20 mg·kg-1 for 14 d. The percentage of MDSCs in bone marrow, spleen, blood, tumor and liver were detected by flow cytometry. Non- esterified fatty acid (NEFA), triglycerides (TG) and total cholesterol (T-CHO) in liver homogenate were tested by a NEFA test kit, a TG test kit and a T- CHO test kit respectively. qRT- PCR and Western blot were used to measure mRNA and protein levels of inflammation-, glycometabolsim- and lipometabolsim-associated indexes in liver. RESULTS MENK decreased percentages of MDSCs in bone marrow, spleen, blood and tumor in colon cancer mice. MENK-treated mice displayed elevated ratio of CD4+T and CD8+T cells in spleen as well as increased T and B lymphocytes proliferation. Meanwhile, MENK also ameliorated liver damage reflected by lower levels of GPT and GOT in serum and reduced risks of cancer- associated index including inflammation, high lipid and high glucose. Furthermore, MENK lowered down the levels of NEFA, TG and T- CHO in liver homogenate. MENK treatment decreased expression of p- STAT3, increased expression of p-AKT, IRS1 and Glut4 at protein level as well as reduced lipogenesis-associated genes and elevated glycolysis-associated genes in liver of tumor bearing mice. Also, abated expression of genes associated with MDSCs generation (M-CSF, GM-CSF, IL-6, IL-1β) and migration (S100A9, KC) was observed within shrunken subcutaneous tumor by MENK intervention. CONCLUSION MENK has the ability to strength immune function against colon cancer by reducing MDSCs and improving liver metabolism.

OBJECTIVE To investigate the effect of phosphotyrosine interaction domain containing 1 (PID1, NYGGF4) on promotion of IR and HCC, and explore its underlying mechanisms. METHODS Lentivirus were used to mediate the knockdown of PID1 in HFD induced IR mouse model as well as ob/ob mice. Intraperitoneal glucose and insulin tolerance were performed 4 weeks after lentivirus injection. Hydrodynamics-based transfection was applied to inducethe liver specific overexpression of PID1. Flow cytometry was exerted to detect the proportion and function of immune cells. qRT-PCR and Western blot were used to detect the expression of downstream pathways of PID1.Immunoprecipitation was used to determine the receptor of PID1. Chromatin immunoprecipitation (ChIP) was operated to measure the modification of H3K4me3 of PID1 promoter. RESULTS PID1 restriction improved insulin resistance, hyperglycemia and fatty liver. Conversely, hepatic knockdown of PID1 attenuated liver xenografted tumor growth. Moreover, PID1 liver- specific protooncogenes via hydrodynamics- based transfection established a primary hepatocellular carcinoma mouse model, induced an immunosuppressive environment, with the reduction of CD3 +, CD4 +, CD8 +T cells, retarded maturation of dendritic cells (DCs), pronounced differentiation of regulatory T cells (Tregs), and recruitment of MDSC. In addition, PID1 overexpression activated proliferation related genes, promoted anti- inflammatory genes, suppressed pro-inflammatory genes, induced glycolysis and lipid metabolism genes to facilitate tumorigenesis in liver. Importantly, PID1 exerted its tumor-promoting function through binding to epidermal growth factor receptor (EGFR) and activation of downstream MAPK pathway. As such, PID1 exist trimethylation of histone H3 at lysine 4 (H3K4me3) modification and IR up-regulated the expression of PID1 by activation the H3K4me3 modification. CONCLUSION PID1 is a new gene that exerts both liver cancer-promoting and insulin resistance inducing function. IR accelerates liver cancer development and progression partially dependent on the activation of PID1.

OBJECTIVETo determine butylidenephthalide in Ligusticum Chuanxiong with RP-HPLC.

METHODThe sample was extracted with methanol using sonication. The ESTD was used to quantify butylidenephthalide. HPLC separation was carried out in a Hypersil ODS columm (4.6 mm x 150 mm, 5 microm) , eluted at 1 mL x min(-1) with methanol-5% isopropyl alcohol (60: 40) at 25 degrees C. The detection wavelength was 230 nm.

RESULTThe linear range was 0.07-0.7 microg for butylidenephthalide. The average recovery was 95.3%, and RSD was 2.3% (n =6).

CONCLUSIONThis method was simple and could be used to determine butylidenephthalide with satisfactory accuracy and reproducibility.

China ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Light ; Ligusticum ; chemistry ; Phthalic Anhydrides ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Rhizome ; chemistry ; Scattering, Radiation

Objective To observe effects of electro-acupuncture (EA) on the expression of apoptosis related protein B-cell lymphoma/leukemia gene 2 (Bcl-2) and Bcl-associated protein X (Bax) in the hippoeampal CA1 region of rats with cerebral infarction,and on their behavior. MethodsAn animal model of cerebral ischemia was established by right side middle cerebral artery occlusion using thread in 48 male,adult Wistar rats,who were then randomly divided equally into an EA group and a control group.Both groups were sub-divided into 1 week,2 weeks and 3 weeks subgroups.The EA group began receiving EA 24h after the occlusion,applied at the Baihui (DU20) and Dazhui (BU14) points,once daily,for one,two or three weeks.The control group was reared conventionally and was not given any special treatment.The rats'learning and memory,motion and neural function were evaluated.The expression of Bcl-2 and Bax in the CA1 region of the hippocampus on the infarcted side and changes of apoptosis indexes were detected using immunohistochemical techniques.ResultsThe learning,memory,motion and neural function of the EA group rats were better on average than those of the control group at all observation time points.The expression of Bcl-2 protein increased along with reduced expression of Bax protein in the EA group significantly more than among the controls.TUNEL positive cells in the CA1 region of the hippocampus were reduced significantly in the EA group compared with the control group.ConclusionsBehavioral ability after cerebral infarction can be improved by EA at the Baihui (DU20) and Dazhui (BU14) points.EA can up-regulate the expression of Bcl-2 and down-regulate the expression of Bax and reduce TUNEL positive cells in the CA1 region of the hippocampus after cerebral infarction,which might be the mechanism of its neuroprotection.

Objective To observe the effect of electroacupuncture combined with physical and hyperbaric therapy on activities of daily living (ADL) of cerebral infarction patients. Methods 105 cases were divided into compositive group (n=35), receiving electroacupuncture combined with physical and hyperbaric therapy; ordinary group (n=35), receiving physical and hyperbaric therapy; and control group (n= 35), not treated. They were assessed with Functional Independence Measure (FIM) and Barthel index (BI) before, 24 d and 36 d after treatment. Results The scores of FIM and BI improved the most in the compositive group (P<0.01) among these groups. Conclusion Electroacupuncture can further improve the activities of daily living of cerebral infarction patients

Objective To investigate the changes of hippocampal neurogenesis and cognitive dysfunction induced by cranial radiation therapy.Methods C57BL/6J mice aged 10 d were subjected to 10 Gy whole brain irradiation with 6 MV X-rays to develop irradiation-induced brain injury model.Morris water maze was designed to estimate spatial learning and memory.At different time post irradiation,brain tissue was removed to stain with hematoxylin-eosin for the pathological results.DCX and PCNA immunohistochemical staining was used to mark the level of neurogenesis in the hippocampus,and ED1immunohistochemical staining to mark the activation of microglia.The TUNEL assay was used to assess the apoptotic neuron death in situ in the hippocampus.Real-time PCR was supplied to inspect the expression of TNF-α and IL-1 β mRNA.Enzyme Linked Immunosorbent Assay (ELISA) was tested for the concentration of TNF-αt in the plasma.Results Pathological studies demonstrated that radiation could induce interstitial edema,inflammatory cell infiltration,cell degeneration,necrosis,apoptosis in the acute phase,edema subsiding,reduction of inflammatory cells,and cytothesis in the dentate gyrus of the hippocampus.IHC studies revealed that,at different time post irradiation,the number of DCX-positive cells and PCNA-positive cells decreased (F =4.9-12.5,5.2-15.7,P ＜ 0.05) but ED1-positive cells increased significantly (F =20.8,P ＜ 0.05).TUNEL-positive cells began to appear in the dentate gyrus of hippocampus 6 h post-irradiation,and its number reached to the highest level at 48 h post-irradiation (F =15.1,P ＜ 0.05).The formation of γ-H2AX foci got at the top 0.5 h post-irradiation (F =18.4,P ＜0.05) and then decreased.After irradiation,the expressions of TNF-α and IL-1β mRNA in the the irradiated group was higher than those of the control group (t =16.3,12.7,P ＜ 0.05).The concentration of TNF-α in the plasma of the irradiated group was higer than that in the control group 3 h post-irradiation,and maximized at 1 week post-irradiation (F =10.5,P ＜ 0.05).Morris water maze tests showed that the latency had no significant differences between the irradiated group and the control group at 1,2,3 d postirradiation,but the latency in the irradiated group was longer than that in the control group with a significant differences at 4,5,6 d post-irradiation (F =7.01,8.17,4.22,P ＜ 0.05).Conclusions Irradiation-induced cognitive dysfunction may be caused by microglial activation and suppression in hippocampal neurogenesis following cranial radiation therapy.

Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.

Objective To explore the effect of electroacupuncture combined with rehabilitation on P300 auditory event-related potentialsin patients with cerebral infarction. Methods 105 patients with cerebral infarction were divided into 3 groups with 35 cases in each group.The comprehensive rehabilitation group received routine rehabilitation and electroacupuncture, the general rehabilitation group only receivedroutine rehabilitation, and the control group received no intervention. They were treated once a day, 10 times as a course. P300 wasdetected before and 3 courses after treatment. Results The latency was shorter and the amplitude was higher in the comprehensive groupthan in the general group (P<0.05). Conclusion Electroacupuncture combined with rehabilitation technology can improve cognitive functionof patients with cerebral infarction.

Objective To investigate the effects of rehabilitation on cognitive function, expression of synaptophysin and neurogranin in rats with cerebral infarction. Methods 48 male Wistar adult rats with cerebral infarction were randomly divided into model group and rehabilitation group, 24 rats in each group. The rehabilitation group received rehabilitation training from the 5th day after modeling. The cognitive ability and the expression of the synaptophysin and neurogranin were observed on the 15th day, 25th day and 35th day after modeling.Results The scores of neural function, motor ability, Y-maze test, and the expression of the synaptophysin and neurogranin were better in the rehabilitation group than in the control group (P<0.05). Conclusion The rehabilitation training can improve the neurological function, motor ability and cognitive ability of rats with cerebral infarction by increasing the positive expression of synaptophysin and neurogranin.

Objective To explore the effect of electroacupuncture combined with rehabilitation technology on the recovery of motor function in patients with cerebral infarction. Methods 105 patients with cerebral infarction were divided into 3 groups with 35 cases in each group. The comprehensive rehabilitation group received routine rehabilitation and electroacupuncture therapy, the general rehabilitation group only received routine rehabilitation, the control group received no intervention. They were treated once a day, 10 times as a course.Fugl-Meyer Assessment (FMA) was used to assess the motor function after 3 courses. Results The scores of FMA were higher in both rehabilitation groups than in the control group (P<0.05), and it was higher in the comprehensive group than in the general group (P<0.05). Conclusion Electroacupuncture combined with modern rehabilitation technology could facilitate to recover motor function in patients with cerebral infarction.