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

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

Objective To detect the expression levels of metallothionein1 H(MT1 H)in children and adoles-cents osteosarcoma serums,and to analyze its relationship with clinicopathological features,and to explore the effect of MT1 H on cell proliferation of osteosarcoma cells and its mechanism.Methods Enzyme -linked immuno sorbent assay (ELISA)was performed to detect the expression of MT1 H in children and adolescents osteosarcoma serums and non-neoplastic disease serums.MT1 H vector was transfected into the osteosarcoma U2OS cells.Reverse transcription -poly-merase chain reaction(RT -PCR)and Western blot were used to detect the expression of the mRNA and protein of MT1 H,respectively.Methylthiazolyldiphenyl -tetrazolium bromide(MTT)was used to detect the cell growth.Western blot was performed to detect the expression of nuclear factor(NF)-κB,and inhibitor of κB (IκB)-αprotein. Results The expressions of MT1 H in osteosarcoma serums and nonneoplastic disease serums was (0.51 ± 0.52)μg/L and (2.17 ±0.78)μg/L,respectively,with a significant difference between the 2 groups(t =-8.966, P <0.05).The expression of MT1 H in stage Ⅰ -ⅡA andⅡB -Ⅲ was (1 .98 ±0.69)μg/L and (2.45 ±0.82)μg/L,respectively,showing a gradual increase depending on clinical staging(t =-2.343,P <0.05).The expressions of MT1 H mRNA and protein were elevated in osteosarcoma U2OS cells after MT1 H vector transfection(all P <0.05). MTT assay showed that,the A value in blank control group,blank vector group,MT1 H vector group were 0.38 ±0.03, 0.36 ±0.03,0.42 ±0.03,respectively,the cell proliferation in the MT1 H vector group was significantly promoted when compared with these in the blank vector group and blank control group(F =4.213,P <0.05)from the third day.West-ern blot showed that,the relative expression of NF -κB in blank control group,blank vector group,MT1 H vector group were 0.56 ±0.05,0.53 ±0.05,0.92 ±0.07,respectively,the relative expression of IκB -αprotein were 0.64 ± 0.06,0.62 ±0.09,0.34 ±0.08,respectively,the expression of NF -κB protein was up -regulated and the expression of IκB -αprotein was down -regulated in the MT1 H vector group when compared with those in the blank vector group and blank control group(F =44.581 ,14.927,all P <0.05).Conclusions The expression of MT1 H is increased in children and adolescents osteosarcoma serums compared with that in nonneoplastic disease serums.The clinical stage is later,the expression of MT1 H is higher.MT1 H promotes cell proliferation through regulating the NF -κB pathway.

Objective To investigate the predictive value of early diffusion-weighted imaging (DWI) for early neurological deterioration (END) after acute isolated basal ganglia infarct.Methods Patients with acute isolated basal ganglia infarct in middle cerebral artery perforators completed head MRI examination at 24 h after onset were analyzed retrospectively.According to the axial DWI revealed lesion size,the maximum diameters were divided into ＜ 15 mm,15-30 mm,and ＞ 30 mm.END was defined as an increase in the National Institute of Health Stroke Scale (NIHSS) score ≥2 or the motor score of NIHSS ≥ 1 at any time within the first 72 h after admission compare with the baseline scores.Results A total of 336 patients were enrolled,including 126 patients (37.5％) with END.There were significant differences in the constituent ratios of the sex (P =0.044),maximum diameter of the lesion (P =0.001),and guilty artery stenosis (P =0.006),as well as baseline NIHSS score (P =0.001),high-density lipoprotein cholesterol (HDL-C) (P =0.033),and C-reactive protein (P =0.039) between the END group and the non-END group.Multiple logistic regression showed that the maximum diameter of the lesions 15-30 mm (odds ratio [OR] 2.360,95％ confidence interval [CI] 1.370-4.066;P =0.002),female (OR 1.660,95％ CI 1.024-2.691;P =0.040),and guilty large artery stenosis (OR 1.713,95％ CI 1.036-2.833;P =0.036) were the independent risk factors for patients occurring END,while the high HDL-C (OR 0.355,95％ CI 0.141-0.894;P =0.028) was an independent protective factor of occurring END.Conclusion Early DWI revealed that the maximum diameter of the lesions may have certain clinical value for prediction of the occurrence of END in patients with acute isolated basal ganglia infarct.

AIM: To investigate the changes of expression of SR-BI in phorbol 12-myristate 13-acetate(PMA) differentiated U937 cells.METHODS: U937 cells were cultured with 100 nmol/L PMA in order to differentiate the cells to macrophages.Immunocytochemical method,Western blotting analysis and reverse transcription polymerase chain reaction(RT-PCR) were used to detect SR-BI protein and mRNA during differentiation.RESULTS: Immunocytochemistry showed that after exposure of U937 cells to PMA for 24,48,72 hours,the values of SR-BI protein expression in U937 cells were 15.94?3.56,27.86?4.39 and 9.08?2.37,with the first two higher than that in undifferentiated cells(7.76?1.74,P0.05) increment in the expression of SR-BI protein compared with U937 monocytes.RT-PCR showed that relative SR-BI mRNA expression in different group was 0.112?0.006,0.235?0.014,0.344?0.140 and 0.138?0.010,respectively.CONCLUSION: SR-BI protein and mRNA were increased after differentiation,reached a peak at 48 hours,and decreased at 72 hours.High expression levels of SR-BI in U937 macrophages following PMA differentiation may be correlated with foam cell formation.

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.