OBJECTIVE: To observe the effect of electroacupuncture (EA) on the rhythm of running-wheel activity of hepatocellular carcinoma (HCC) mice and the expression of Per 1 and Per 2 (circadian rhythm genes) in the hypothalamic suprachiasmatic nucleus (SCN), so as to investigate its mechanism underlying regulating circadian rhythm. METHODS: A total of 108 male C 57 BL / 6 J mice were randomly divided into control, HCC model and EA groups which were further assigned to six zeitbeger (environmental light-dark cycle) time (ZT) point (ZT 0, ZT 4, ZT 8, ZT 12, ZT 16 and ZT 20) subgroups. The HCC model was established by injection of H 22 cancer cell (abdominal 3rd generation, 10 µL) suspension into the larger live lobe. Mice of the control group received saline injection of the liver lobe. EA (2 Hz/15 Hz, 0.2 mA) was applied to bilateral "Ganshu" (BL 18) and "Zhiyang" (GV 9) for 15 min, once daily for 10 days. Mice of the control and model groups received the same binding-fixing to those of the EA group. Circadian running-wheel activity of 12 h∶12 h light darkness (LD) cycle (activity onset and acrophase of actogram, amplitude or peak of periodogram) was recorded by using ClockLab (ACT-500) software and analyzed by MATLAB (R 2007 b) before and after EA treatment. The pathological changes of liver cells were observed under light microscope after sectioning and H．E. staining. The expression levels of Per 1 mRNA and Per 2 mRNA in the liver tissues were determined by fluorogenic quantitative real time-PCR. RESULTS: (1) Following modeling, the amplitude of periodogram of running-wheel activity was significantly lowered at ZT 0, ZT 4, ZT 8, ZT 12, ZT 16, and ZT 20 relevant to the control group (P<0.05). After EA intervention, the amplitude of periodogram at ZT 8 (15：00) was considerably increased relevant to the model group (P<0.05), and the acrophase at ZT 8 was remarkably advanced (P<0.05). No significant changes were found in the onset time and periods of periodogram at the 6 time-points after modeling and EA intervention (P>0.05). (2) The expression levels of Per 1 mRNA and Per 2 mRNA in the SCN were significantly up-regulated at the 6 time-points in the model group relevant to the control group (P<0.05), and obviously down-regulated at ZT 8 after EA intervention relevant to the model group (P<0.05)．. CONCLUSION: EA can benignly regulate the rhythm of running-wheel activity of HCC mice, which may be closely related to its effect in down-regulating the expression of circadian rhythm genes Per 1 and Per 2 in the SCN.

Objective: To explore the material basis and anti-inflammatory mechanisms of Zukamu granules (ZKMG)based on the headspace-solid phase microextraction-gas chromatography-mass spectrometry(HS-SPME-GCMS) technique and network pharmacology analyses. Methods: HS-SPME-GC-MS technique was used to extract and identify the volatile components of ZKMG. Then combined with the non-volatile components obtained by literature retrieval, we constructed the main chemical composition table. The bioactive components were screened based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and their targets were predicted by the Swiss Target Prediction database. Thereafter, the anti-inflammatory-related targets were screened based on the Thera- peutic Targets Datebase and DrugBank with”Anti-inflammatory”as keyword. Then, the STRING database to analyze protein-protein interaction(PPI)and the software of Cytoscape 3.6.1 was used to construct the component-target-antiinflammatory target network. Finally, enrichment of function and signaling pathways of the target genes was conducted by Gene Ontology(GO)database and the Database for Annotation, Visualization and Integrated Discovery(DAVID). Re- sults: Thirty bioactive components including thymol, kaempferol and quercetin in ZKMG were screened out. Ninety-seven targets were predicted and AKR1B1, ALOX15, ALOX5, CYP1A2, EGFR, CHRM1, NOS3, PLA2G1B, PTGS2, BCHE were key anti-inflammatory targets. Sixty-eight pathways related to anti-inflammation were obtained by KEGG enrichment analysis, mainly including arachidonic acid metabolism, MAPK signaling pathway, inflammatory mediator regulation of TRP channels, IL-17 signaling pathway and NF-κB signaling pathway. Conclusion: This study focuses on the chemical constituents of ZKMG and network pharmacology to predict the material basis and pharmacological mecha- nism of their anti-inflammatory effect, which lays a good foundation for the verification of the later biological experiments and provides certain reference basis for the research on the anti-inflammation mechanism of other compound preparations.

Objective To study the relevance of 1iver fibrosis at Kasai procedure with age, degree of pathological changes of liver tissue, header bile ductuler proliferation and pathological feature of portal fibrous tissue in biliary atresia patients. Methods From January 2013 to June 2016, 48 biliary atresia patients accepted Kasai procedure, and liver tissue biopsy and portal fibrous tissue excision was done. Level of liver fibrosis was evaluated under microscope using Masson trichrome staining and HE staining, and extent of ductular reaction was evaluated using CKl9 immunohistochemistry staining with image analysis technology. Based on classification of Okamoto severity degree of liver fibrosis, forty eight patients were divided into two groups:low-grade liver fibrosis group (23 patients)and severe liver fibrosis group (25 patients). The relevance of age at Kasai procedure, liver pathological changes, header bile ductuler proliferation, portal fibrous tissue in volume and bile ductuler proliferation, inflammation reaction with severity degree of liver fibrosis was analyzed with SPSSl6.0 software statistically. Results Age of severe 1iver fibrosis group at Kasai procedure were 38- 89 d, and the mean age was (62.64 ± 14.92) d. Age of low-grade 1iver fibrosis group were 18- 92 d, and the mean age was (53.13 ± 16.89) d. There was significant difference (t=2.071, P=0.044). Mean number of patients with header bile ductuler proliferation at Kasai procedure in severe 1iver fibrosis group and low-grade 1iver fibrosis group were (17.28 ± 4.31) cases and (11.39 ± 4.00) cases, and there was significant difference (t=4.900, P=0.000). There was significant difference in degree of hepatocellular pathological changes at Kasai procedure between groups (χ2 =4.691, P=0.030). There were no significant differences in the volume of portal fibrous tissue in volume (t=0.276, P=0.784), portal bile ducts proliferation (t=0.53, P=0.60) and inflammation reaction (χ2=3.074, P=0.080) between groups. Conclusions There is significant relevance between the degree of 1iver fibrosis in biliary atresia patients at Kasai procedure and age, level of hepatocellular pathological change and degree of header bile ductuler proliferation , The more older the age, the more sever the hepatocellular pathological change and the bile ductuler proliferation, the more sever the liver fibrous.

Objective To investigate the effect and safety of self-made blenderized diet used for nutritional support treatment in the ICU critically ill patients with diabetes. Methods 74 ICU critically ill patients with diabetes were randomly divided into two groups:study group and control group. The self-made blenderized diet were given to the patients in study group and enteral nutrition to the patients in control group for nutritional support treatment for four weeks. Then glucose metabolism, various nutrition indicators and gastrointestinal dysfunction were observed in these two groups. Results The fasting blood glucose and fructosamine levels were decreased significantly,while the nutritional indicators was significantly higher after treatment than at baseline in all patients. There was no significant difference in the levels of fasting blood glucose, postprandial glucose, fructosamine and nutritional indicators between two groups after treatment,so were the incidence of the hypoglycemia and gastrointestinal complications. Conclusion The self-made blenderized diet was a good enteral nutrition support therapy for ICU critically ill patients with diabetes which had a considerable effect, but more cheaper,and practical compared to the enteral nutrition emulsions.

OBJECTIVETo explore the performance patterns of reaction point Jingtong in balance acupuncture through multi-center and big-sample clinical investigation. Methods The Jingtong points of balance acupuncture on healthy side and affected side were observed among 230 cases of cervical spondylosis and scores of self-discomfort in reaction point, color of skin, changes of skin, morphology of subcutaneous tissue and abnormal pressing pain were recorded. The software SPSS 15.0 was applied to statistically analyze the recorded scores.

RESULTSAmong 230 cases, the reaction point appeared in 226 cases, accounting for 98. 3%. Among the 226 cases who had reaction point, the total score of symptom and sign was (1.08+/-1.09) on the healthy side and (0. 84+/-1. 36) on the affected side, which had statistical significance (P<0. 01); score of self-discomfort in reaction point was (0. 76 +/-0. 83) on the healthy side and (0. 40+/-0.80) on the affected side, which had statistical significance (P<0.01); the score of skin color was (0.10+/-0.36) on the healthy side and (0. 03+/- 0. 19) on the affected side, which had statistical significance (P<0. 05); the score of abnormal pressing pain was (2. 47+/-2. 46) on the healthy side and (1. 39+/-2. 37) on the affected side, which had statistical significance (P<0. 01).

CONCLUSIONThe total score of symptom and sign of reaction point Jingtong on the healthy side is higher than that on the affected side, indicating positive reaction of Jingtong on the healthy side has specificity for cervical spondylosis. When patient has cervical spondylosis on either side of neck, the other side will have anomaly in Jingtong.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Cross-Sectional Studies ; Female ; Humans ; Male ; Middle Aged ; Neck Pain ; therapy ; Spondylosis ; therapy ; Young Adult

This study is to evaluate the effects of the metformin (Met) on β cell function of diabetic KKAy mice. Female diabetic KKAy mice selected by insulin tolerance test (ITT) were divided randomly into two groups. Con group was orally administered by gavage with water, Met group with metformin hydrochloride at a dose of 0.2 g x kg(-1) for about 12 weeks. ITT and glucose tolerance tests (OGTT) were determined. Beta cell function was assessed by hyperglycemic clamp. Pancreatic biochemical indicators were tested. The changes of gene and protein expression in the pancreas and islets were also analyzed by Real-Time-PCR and immunostaining. Met significantly improved glucose intolerance and insulin resistance in KKAy mice. Fasting plasma glucose and insulin levels were also decreased. In addition, Met markedly increased glucose infusion rate (GIR) and elevated the Ist phase and maximum insulin secretion during clamp. It showed that Met decreased TG content and iNOS activities and increased Ca(2+) -Mg(2+)-ATPase activity in pancreas. Islets periphery was improved, and down-regulation of glucagon and up-regulated insulin protein expressions were found after Met treatment. Pancreatic mRNA expressions of inflammation factors including TLR4, NF-κB, JNK, IL-6 and TNF-α were down-regulated, p-NF-κB p65 protein levels also down-regulated by Met. And mRNA expressions of ion homeostasis involved in insulin secretion including SERCA2 and Kir6.2 were up-regulated by Met. Met increased SIRT5 expression level in pancreas of KKAy mice under the hyperglycemic clamp. These results indicated that chronic administration of Met regulated pancreatic inflammation generation, ion and hormone homeostasis and improved β cell function of diabetic KKAy mice.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Down-Regulation ; Female ; Glucose Tolerance Test ; Homeostasis ; Inflammation ; drug therapy ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Interleukin-6 ; metabolism ; Metformin ; pharmacology ; Mice ; NF-kappa B ; metabolism ; Pancreas ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the effect of Mudan Granule (MD) on the glucose metabolism and beta cell function in monosodium glutamate (MSG) induced obese mice with insulin resistance (IR).

METHODSMSG obese mice were induced by subcutaneous injecting MSG (4 g/kg for 7 successive days in neonatal ICR mice). Forty MSG mice with IR features were recruited and divided into four groups according to body weight, fasting blood glucose, triglyceride (TG), total cholesterol (TC), and the percentage of blood glucose decreased within 40 min in the IR test, i.e., the model group (Con), the low dose MD group, the high dose MD group, and the Metformin group (Met). Besides, another 10 ICR mice were recruited as the normal control group (Nor). The water solvent of 2.5 g/kg MD or 5 g/kg MD was respectively administered to mice in the low dose MD group and the high dose MD group. Metformin hydrochloride was given to mice in the Met group at 0.2 g/kg body weight. Equal dose solvent distilled water was administered to mice in the Nor group and the Con group by gastrogavage, once per day. All medication was lasted for 15 weeks. Insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed after 6 weeks of treatment. Beta cell function was assessed by hyperglycemic clamp technique. The morphological changes in the pancreas were evaluated by hematoxylin-eosin (HE) staining. Changes of iNOS, NF-kappaB p65, and p-NF-kappaB p65 in the pancreas were tested.

RESULTSCompared with the Nor group, the blood glucose level, AUC, and fasting blood insulin, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, pNF-kappaB p65 subunit obviously increased; decreased percentage of blood glucose within 40 min in ITT, glucose infusion rate (GIR), Clamp 1 min insulin, and Max-Insulin obviously decreased in the Con group (P < 0.05, P < 0.01). Compared with the Con group, the aforesaid indices could be improved in the Met group (P < 0.05, P < 0.01). In the low dose MD group, AUC, iNOS activities, and the expression of iNOS and p-NF-kappaB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT and GIR obviously increased (P < 0.05, P < 0.01). In the high dose MD group, AUC, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, and p-NF-KB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT, Max-Insulin, and GIR obviously increased (P < 0.05, P < 0.01).

CONCLUSIONMD could significantly improve IR and functional disorder of 3 cells in MSG obese mice, which might be associated with lowering inflammatory reaction in the pancreas.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; metabolism ; Male ; Metformin ; pharmacology ; Mice ; Mice, Inbred ICR ; Mice, Obese ; Obesity ; chemically induced ; metabolism ; Pancreas ; cytology ; drug effects ; Sodium Glutamate

We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.

Objective To analyze the efficacy,late complications and prognostic factors of post-op-erative radiotherapy for cervical cancer. Methods From Nov. 1999 to Feb. 2005,114 patients with cervi-cal cancer received adjuvant pelvic radiotherapy after radical hysterectomy and pelvic lymphadenectomy. The median age was 42.5 (24 to 72) years old. According to the FIGO staging system,6,51,18,26 and 13 pa-tients had stage ⅠA, Ⅰb1, Ⅰb2, ⅡA and ⅡBdisease. The pathological type was squamous cell carcinoma,ade-nocarcinoma, squamous-adenocarcinoma and undifferentiated carcinoma in 92,19,2 and 1 patients, respec-tively. The whole-pelvic external beam irradiation of 50 Gy (40 to 60 Gy) was given with 6 MV or 15 MV X-ray beams using four-field box technique. Eighty-one patients received intravaginal brachytherapy of 16 Gy (4 -30 Gy in 1 -6 fractions) 4 weeks after the beginning of radiotherapy, with the referrence point being at 0.5 cm under the vaginal mucosa. Eighty-seven patients received preoperative and/or concurrent chemother-apy. The survival and independent prognostic factors were analyzed by Log-rank method and Cox model. Results The median follow-up time was 26.0 (5 - 75) months. The overall survival rate, disease-free sur-rival rate and local control rate were 93.1%, 88.1% and 94.6% at 2-year, and 75.7%, 62.3% and 85.6% at 5-year,respectively. The independent prognostic factors were lymph node metastasis and positive surgical margin for overall survival, positive surgical margin for local control, and stage, uterine body invasion and positive surgical margin for disease-free survival. Sixteen patients ( 14% ) had distant metastasis, and the most common sites were the lung,inguinal region,bone,liver and brain. According to RTOG grading sys-tem, the incidence of late gastrointestinal side effects of grade 1,2 and 3 was 11.4%, 11.4% and 3.5%. The corresponding genitourinary side effects were 14.0% ,6.1% and 0.9%, respectively. The incidence of leg lymphedema was 7 % . Conclusions Post - operative radiotherapy can achieve good local control in cervical cancer with acceptable late side effects. Distant metastasis is the main cause of death.

Objective To analyze the outcomes of radiotherapy for 74 patients with intracranial ger-minoma. Methods Between 1990 and 2007,74 patients with intracranial germinoma(9 pathologically diag-nosed and 65 clinically diagnosed) were treated with radiotherapy in our hospital. The median age at diagno-sis was 15 (range 5-45) years. Radiation treatment fields varied among patients, including craniospinal irra-diation(CSI), whole brain irradiation, whole ventricular irradiation with primary tumor boost, and involved-field irradiation only to the primary tumor plus margin. The dose was 38.5 -50.0 Gy to the tumor,18-25 Gy to the whole brain/ventricular,and 21-25 Gy to the whole spinal cord in fractions of 1.6-2.0 Gy per day,5 fractions per week. Results The median follow-up time was 80(range 12-168) months and the fol-low-up rate was 97%. Fourteen patients had been followed up for over 10 years. The 1-,5- and 10-year o-verall survival rates were 99% ,96% and 93%. The corresponding disease free survival rates were 97%, 90% and 83%, respectively. Relapses occurred in 9 patients. For the 6 patients with in-field relapse, the dose to the tumor was 38.5-40.0 Gy in 3 patients, 41-45 Gy in 2 and 46-50 Gy in 1. Relapse in the spinal cord was found in 3 patients and none of them received spinal irradiation. Twenty-one patients re-quired hormonal replacement therapy because of radiation induced hypofunction of prehypophysis. Conclu-sions Radiotherapy alone is a curative treatment for intracranial germinoma. The proper dose should be de-termined by tumor numbers and the examination of cerebrospinal fluid.