Several M-CAT elements are present in the chicken nicotinic acetylcholine receptor γ-subunit promoter and required for the totipotency of this gene.To understand the binding properties of M-CAT element,the interaction of M-CAT motif,at the position -67/-61 relative to the initiation site of γ-promoter with nuclear proteins,were analyzed.Mutageneses and binding experiments demonstrated that the M-CAT core motif bound nuclear protein (s) from tested chicken tissues to generate a high-mobility complex;while the flanking sequence bound different factors to generate lower-mobility complexes with the exception of liver.Southwestern blotting showed that only a 30 kD protein was expressed in tested tissues of chicken embryo at day 13.These results suggest that a 30 kD factor can directly bind to M-CAT element in the manner of monomer,even homodimer and is ubiquitously expressed in tissues,while the binding of the factor(s) to flanking sequence should probably rely on the presence of a 30 kD ubiquitous factor.

Acupuncture Therapy ; Humans ; Meridians ; Muscles ; physiopathology ; Nervous System Diseases ; physiopathology ; therapy

Objective To detect the osteopontin(OPN)expression in renal tissue of rats with fluorosis and low calcium diet,and study the role of OPN in renal injury of fluorosis.Methods Forty-eight aged 1 month Wistar rats,80-120 g,were randomly divided into 4 groups by 2×2 factorial design(the number of female and male in each group was equal):the control group,high-flluoride group,low-calcium group and low-calcium with high-fluoride group.All rats of the fluorosis groups were fed with feed containing corn exposed to coal-burning from endemic fluorosis areas with high fluoride(100 mg/kg,corn),the other two groups were fed with feed containing coru from nonendemic fluorosis areas(fluoride 5 mg/kg,corn).After 16 weeks,the rats were killed.The change of teeth was examined,and the incidence rate of dental fluorosis was calculated.The expressions of both protein and mRNA of OPN in rat renal tissue were determined by RT-PCR and immunohistochemistry after four-month experimentation.Results The growth of teeth was very well in the control group and the low-calcium group.The two high-fluoride groups showed evident dental fluorosis(100%).The results of immunohistochemistry showed that the OPN protein was localized in renal tubule cytoplasm.The OPN-positive cells from renal tissue were lightly and scatteredly stained in control and low-calcium groups.The OPN-positive cells had deeper color in high-fluoride group and low-calcium with high-fluoride group,widely distributed in the renal tubular epithelial cells.The protein expression of OPN in the two groups exposed to fluoride(168.64±13.21,169.26±8.92)was significantly higher than those of the corresponding control group(145.78±10.26,all P<0.01)and low-calcium group(149.60±16.84,all P<0.01).The mRNA expression of OPN in the two groups exposed to fluoride(1.89±0.37,1.94±0.22)was significantly higher than those of the corresponding control group(1.32±0.26,all P<0.05)and low-calcium group(1.30±0.186,P<0.05),respectively.High fluoride influenced the expression of protein and mRNA of OPN(F=13.821,4.24,all P<0.05).Low calcium did not affect the expression of protein and mRNA of OPN(F=2.164,0.58,all P>0.05).However,high fluoride and low calcium had a cross interaction on the expression of protein and mRNA of OPN(F=6.257,432,all P<0.05).Conclusions Over-dose fluoride enhances the expression of OPN.The higher expression observed in the cases exposed to high fluoride concentration is associated with serious renal injury.OPN may he a potential marker for renal injury in fluorosis.Moreover,over-dose fluoride and low calcium make the renal injury worse,indicating low calcium plays an important part in renal injury by fluoride.

Objective To investigate the protective effects of total rhizoma panacis japonica saponins (tRPJS)on rat brains after focal cerebral ischemia injury, and to explore its protective mechanisms of tRPJS. Methods Focal cerebral ischemia in rats was produced by 24 h occlusion the right part of the middle cerebral artery(MCAO).The activities of glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA)in ischemic tissue of brain as well as the brain pathological changes were analyzed. Results The activities of GSH-PX, SOD and CAT were lowered and the content of MDA was elevated in the model rats. Treatment with tRPJS 200, 100, 50 mg? kg-1 could significantly improve the activities of all the enzymes mentioned above and reduce the content of MDA in ischemic tissue of brain. Conclusion The tRPJS has protective effect against focal cerebral ischemic injury and this may be related to the increase of activities the GSH-PX, SOD and CAT and to the reduction of the lipid peroxidation.

Objective To investigate the expression of hypoxia-inducible 1 alpha(HIF-lα)and glucose transporter 1(Glut1)in human breast cancer and its relationship to proliferating cell nuclear antigen (PCNA)protein and clinical pathologic factors.Methods Immunohistochemical staining was used to measure the expression of HIF-lα.Glut1 and PCNA in human breast fibroadenoma,usual hyperplasia and breast carcinoma.Results HIF-1α expression was not found in breast fibroadenoma and hyperplastic Iesions.In contrast.the positive rate of HIF-1α was found in the ductal carcinoma in situ 55%(DCIS,11/20)and the invasive breast carcinoma 85%(51/60).Glut1 positivity in breast carcinoma was 58.8%(47/80).The totsl positive rate of PCNA in breast carcinoma was 75%(60/80),that in DCIS was 65%(13/20)and that in invasive carcinoma was 78.3%(47/60).There was a positive correlation between HIF-lα and Glut 1 level (r=0.653,P＜0.01),a positive correlation between HIF-1α and PCNA level(r=0.693,P＜0.01);and also a positive correlation between Glutl and PCNA level(t=0.742.P＜0.01).conclusion The overexpression of HIF-lα and its target gene Glut1 played important roles in carcinogenesis and progression of breast carcinoma and closely correlated with cell proliferation of breast carcinoma and may become a new target for treatment of breast carcinoma.

OBJECTIVETo evaluate preliminarily the efficacy on functional constipation treated with electroacupuncture of different acupoint prescriptions.

METHODSOne hundred and four patients were randomized into a front-mu and back-shu points group (19 cases), a he-sea points group (34 cases), a he-sea, front-mu and back-shu points group (26 cases) and a western medication control group (25 cases). In the front-mu and back-shu points group, electroacupuncture was applied at bilateral Tianshu (ST 25) and Dachangshu (BL 25). In the he-sea points group, electroacupuncture was applied at bilateral Quchi (LI 11) and Shangjuxu (ST 37). In the he-sea, front-mu and back-shu points group, electroacupuncture was applied at unilateral Tianshu (ST 25), Dachangshu (BL 25), Quchi (LI 11) and Shangjuxu (ST 37). In the three groups above, the treatment was given 5 times a week in the first two weeks and 3 times a week in the next two weeks. In the western medication control group, mosapride citrate tablets were prescribed for oral administration, 1 table (5 mg) each time, 3 times a day, continuously for 4 weeks. The period of research was 9 weeks, including 1 week for baseline evaluation, 4 weeks for treatment and 4 weeks for follow-up. The weekly defecation frequency was taken as primary index, while the defecation difficulty and life quality score were taken as the secondary indices for the efficacy evaluation after treatment and in follow-up.

RESULTSAccording to the intention-to-treat (ITT) analytic principle, 104 cases were all enrolled in the final analysis. (1) After treatment, the weekly frequency of defecation was all increased significantly in the four groups (P < 0.05, P < 0.01). The efficacy of the three electroacupuncture groups was similar to that of western medication control group (P > 0.05). In follow-up, the increasing effect on the weekly frequency of defecation was maintained in the he-sea points group (P < 0.01), superior to the front-mu and back-shu points group and the western medication control group (P < 0.05, P < 0.01); the weekly frequency of defecation was not improved in the rest three groups (P > 0.05). (2) After treatment, defecation difficulty was relieved in the he-sea points group, the he-sea, front-mu and back-shu points group and the western medication control group (P < 0.05, P < 0.01). In follow-up, the improvements were still significant in the he-sea points group and the he-sea, front-mu and back-shu points group (both P < 0.01). (3) After treatment, the life quality score was significantly improved in the patients of the he-sea points group (P < 0.05). The difference was not significant in the rest three groups as compared with that before treatment (all P > 0.05).

CONCLUSIONThe weekly frequency of defecation is increased effectively after treatment in the three electroacupuncture groups and the efficacy is similar to mosapride citrate tablets. The bilateral Quchi (LI 11) and Shangjuxu (ST 37) in he-sea acupoints increase significantly the weekly frequency of defecation, relieve defecation difficulty and improve life quality. Acupuncture efficacy is sustained for 4 weeks. This acupoints prescription is the best in the treatment of functional constipation.

Acupuncture Points ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

The COI gene as DNA barcode was used to identify the Manis pentadactyla and its adulterants in order to provide a scientific basis for the molecular identification of M. pentadactyla. Genomic DNA was extracted from experimental samples using the DNA extraction kit. The COI genes were amplified using polymerase chain reaction (PCR) and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. The neighbor-joining (NJ) tree was constructed by MEGA 6.0. The results indicated that COI sequences were successfully amplified and NJ trees results indicated that M. pentadactyla and its adulterants can be easily identification. Therefore, the COI gene is an efficient barcode for identification of M. pentadactyla and its adulterants,which will provide a new technique for the market supervision.
Animals ; Cattle ; DNA Barcoding, Taxonomic ; methods ; Drug Contamination ; prevention & control ; Electron Transport Complex IV ; genetics ; Mammals ; classification ; genetics ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Sheep ; Swine

Objective To compare the saccharometabolism with the pancreatic islets functions and insulin resistance index in children with severe stress. Methods Thirty children with severe stress and 30 healthy children in control group were tested. The levels of fasting blood glucose (FBG), fasting insulin (FINS) and fasting C - peptide (FCP) were detected by radioimmunoassay respectively and insulin sensitivity index (ISI), insulin resistance index (IR) and fasting blood cell function index (FBCI) were calculated statistically. Results There were significant differences between the children with severe stress and the normal controls in the levels of FINS, FCP and FBG,(all P0.05). Conclusion There is insulin resistance with the significant decrease in the insulin sensitivity index and significant increase in insulin resistance index in the children with severe stress, which may cause the disorder in glucose metabolism in children with severe stress.

Objective To investigate the meaning of PURA gene and its protein in nephridial tissue of the rats with endemic fluorosis of coal burning. Methods Thirty-six SD rats of 80 - 100 g, body weight were randomly divided into control group, low fluorosis group and high fluorosis group according to body weight, 12 in each group, the number of female and male in each group was the same respectively. The control group, Low fluorosis group and high fluorosis group rots were fed with 1.5,25.0,60.0 mg/kg fluoride content in feedstuff, to establish the animal model of fluorosis. Expressions of both mRNA and its protein of PURA gene in rat nephridium tissue, were determined by RT-PCR and immunohistochemistry after four-month experimental period. Results The expressions of PURA mRNA[(2.74± 1.06),(4.29 ± 2.11)] and its protein[ (28 827.91 ± 4801.94),(61 146.96 ± 4997.55)] in low fluorosis group and high fluorosis group was higher than that in the control group[ ( 1.13 ± 0.87), (7131.95 ± 1524.54), all P < 0.05]. And the expressions of PURA mRNA and protein in high fluorosis groups was higher than that in low fluorosis greup(all P < 0.05). Conclusion High fluoride can lead to the high expression of PURA gene mRNA and protein in the rat nephridium tissue exposed to sodium fluoride.

Objective To explore the relation between the expression of PBMCs IP-10 mRNA and systemic lupus erythematosus.Methods The expression of PBMCs IP-10 mRNA was investigated by RT-PCR semi quantitative method and samples from 46 patients with SLE,20 patients with RA,11 non-SLE patients with renal impairment and 20 healthy volunteers.Results The expression of PBMCs IP-10 mRNA in active SLE group was significantly higher than that in inactive group(P0.05).Serum levels of IP-10 were highly correlated with the expression levels of PBMCs IP-10 mRNA(r=0.897 1,P