Objective To observe the effects of auricular point plaster therapeutic time on pain caused by cyclomastopathy .Methods 90 patients who visited breast surgery outpatient clinic and were diagnosed as cyclomas -topathy due to stagnation of liver -QI were randomly divided into the three groups by numerical table:3 days ( group A),5 days(group B),7 days(group C) ear acupoint therapy group,30 cases each.Improved pain symptoms of cyclo-mastopathy were compared among the three groups .Results There was statistical significance to reduce pain symp-toms in the three groups[before the treatment:(4.57 ±1.74)points,(4.63 ±1.83)points,(4.81 ±1.57)points;after the treatment:(3.45 ±1.52)points,(1.82 ±0.96)points,(2.45 ±1.03)points.t=2.66,7.45,6.88,all P<0.05].There was obvious statistical difference between group B and group C (t=7.45,6.88,all P<0.01),and low recurrence rate .Conclusion 5 days auricular point plaster therapy is worthy of clinical popularization because which can improve pain symptom of cyclomastopathy ,shorten time of therapy with high compliance and low recurrence rate .

Objective To study the mechanism of activity change of gastric alcohol dehydrogenases(ADH).We investigated ADH before and after ingestion of alcohol on rats.Methods Thirty-nine rats were divided into two groups at random:model ones(24 cases) and control ones(15 cases).Model group of rats were infused into stomaches by alcohol,and control ones by distill water.Structures of cell and sub-cell in gastric mucous membrane were observed by enzyme electron microscope and hematoxylin eosin(HE) stain.Results It was shown that there were dropsical degeneration of chief cells and parietal cells of stomaches by HE stain,and the mitochondria of parietal cells and the rough-surfaced endoplasmic reticulum(rER) of chief cells were destroyed by enzyme electron microscope in model groups after two months of ethanol perfusion.There were not change in control group.Conclusions The mechanism of activity alterations of gastric ADH may result from dropsical degeneration of chief cells and parietal cells of stomach,which result in degeneration,solution and breakage of construction of rER in the chief cells and mitochondria in parietal cells.

Purpose: To observe the long-term and recent clinical effect of needle-warming method plus functional training in the treatment of osteoarthritis of the knee joint. Methods:After 106 cases of the patients were divided into the needle-warming group and comprehensive group by the order of their first visit, the needle-warming group was treated by the needle-wanning method and the comprehensive group was treated by the needle-warming method plus functional training. After ten treatments, the clinical data in the two groups were evaluated upon Lequesne scale before and after the treatments. Three months later, follow-up survey was given to process the statistic management of the reoccurrence rate of the symptoms.Results: In the two groups after the treatment, the symptoms of the knee joint and daily life ability were obviously improved than the respective conditions before the treatment, and the integral decreased (P＜0.05), with no significant difference (P＞0.05) between the two groups. In the follow-up survey, there was a difference (P＜0.05) between the two groups in the positive rate of the recurrent symptoms. Conclusion: Certain therapeutic effect exists in the treatment of osteoarthritis of the knee joint by the needle-warming method. If functional training is combined,the therapeutic effect would be more stable, without easy reoccurrence.

AIM: To develope a reversed-phase high performance liquid chromatographic method for the quantitative determination of chlorogenic acid and caffeic acid in Shegan Kangbingdu Injection. METHODS: The operation was carried out in the Kromasil ODS column with the mobile phase consisting of a mixture of water-methanol-acetonitril aceti acid(95∶10∶5∶2,v/v),in which the flow rate of 1.0 mL/min and UV detection wavelength at 326 nm were set to determine the contents of chlorogenic acid and caffeic acid. RESULTS: There were good linear relations between the concentrations and the peak-areas of chlorogenic acid and caffeic acid.The relative standard deviation of peak areas for chlorogenic acid or caffeic acid were 0.60%,0.32%,respectively.The two kinds of standard solutions were both stable in 16 h(RSD=0.54% for CGA,0.11% for CFA).The average recovery was 100.1%,99.6% for CGA,CFA,repectively.The detectable limit(S/N=3)was 0.012,0.020 mg/L,and the quantitative limit (S/N=10) was 0.037,0.052 mg/L,repectively. CONCLUSION:The method is simple,sensitive,rapid and accurate,and can be used for the quality control of Shegan Kangbingdu Injection

Objective To establish a double reporter transgenic mice for monitoring Cre recombinase activity. Methods ZAP DNA fragment with lacZ and human alkaline phosphatase (hAP) gene was microinjected into the male pronucleus of 554 fertilized eggs from C57BL/6 mice. The founder mice and their progeny were screened for integration of transgene into the mouse genome by PCR and Southern blotting. The expression of lacZ transgene at early embryos from F1 generation mice was analyzed by X gal staining. Results A total of 398 survival ZAP DNA injected fertilized eggs were transfered to the oviducts of 21 pseudopregnant recipient mice. Of the 21 recipient mice, 13 became pregnancy and gave birth to 68 offspring mice. The zygote survival rate and birth rate were 71% (398/554) and 17% (68/398), respectively. Of the 68 offspring mice, 9 mice (5 males and 4 females) were identified by PCR and Southern blot analysis. Total integration rate and efficiency of transgene was 13% (9/68) and 1.6% (9/554), respectively. Nine mice as the founders were back crossed to set up F1 generation with other inbred C57BL/6 mice. Out of 9 transgenic mice, transmission of reporter gene in F1 offspring mice followed Mendelian rules, but the expression of lacZ protein was detected at the early embryonic stage (13.5 days postcoitum) in only 3 mice. Conclusion A double reporter transgenic mice for monitoring Cre recombinase activity is established.

Objective To synthesize novel gold nanoparticles of GAL-PEG-GNPs,study its radiation effect on hepatocellular carcinoma cells HepG2 cells in vitro,and investigate the underlying mechanisms.Methods GAL-PEG-GNPs were synthesized and characterized successfully.HepG2 cells were divided into three groups of control,GNPs and GAL-PEG-GNPs.The cytotoxicities of these compounds were tested by the CCK-8 assay and their IC50 values of HepG2 cells were calculated.Cell uptake of nanoparticles was detected by TEM and ICP-MS.The radiosensitization effect of nanoparticles was tested by the colony formation assay.Cell cycle distribution was detected by FCM.The expressions of CAT,SOD,and total GSH were detected with a microplate reader,and the expressions of apoptosis-related proteins were tested by Western blot.Results The GNPs and GAL-PEG-GNPs had absorption peaks at 520 and 530 nm,respectively,and their diameters were (22.6-±2.12) and (32.0 ± 1.41) nm detected by ICP-MS.The GAL-PEG-GNPs and GNPs had similar cytotoxicity profiles (P ＞ 0.05),while GAL-PEG-GNPs could be more effectively uptaken by HepG2 cells than GNPs.The sensitive enhancement ratio (SER) of GNPs and GAL-PEG-GNPs to HepG2 cells were 1.46 和 1.95,respectively.The percentage of cells at phase of G2/M in HepG2 population treated with GNP was higher than that of untreated cells (t =14.20,P ＜0.05).The protein expressions of Cytochrome C,Bax,Caspase-3,and Caspase-9 were upregulated while Bcl-2 expression was down-regulated in the cells treated with GNPs/radiation or GAL-PEGGNPs/radiation.The expressions of CAT,SOD and total GSH in the GNP treated groups were significantly decreased compared with the control group(t =12.34,29.39,12.85,P ＜ 0.05).Conclusions GALPEG-GNPs has obvious radiosensitization effect on HepG2 cells,which is related to the induction of cell apoptosis and the generation of free radicals.

Objective:To analyze long-term therapeutic effect and safety of warfarin anticoagulant therapy of differ- ent intensity on aged patients with nonvalvular atrial fibrillation (NVAF). Methods:According to age,a total of 197 NVAF patients followed up for five years were divided into advanced aged group [n=65,≥80 (85.00±2.09) years],aged group [n=75,65-79 (76.50±2.27)years]and middle-aged group [n=57,<65 (57.40±2.18) years].All enrolled patients received long-term warfarin anticoagulant therapy,advanced aged group and aged group received low intensity anticoagulation,international normalized ratio (INR)was 1.6~2.5,while middle-aged group received standard intensity anticoagulation and the INR was 2.0~3.0. Thrombus events and incidence rates of hemorrhage etc.over five years were compared among three groups,and the safe dose range of warfarin was ex- plored.Results:During five-year follow-up,no acute cerebral infarction occurred in three groups.The bleeding and other adverse reaction among three groups were no significant difference (P>0.05).Compared with middle- aged group,there were significant reductions in warfarin dose [(3.29±0.49)mg/d vs.(2.95±0.38)mg/d,(2.85 ±0.49)mg/d],INR [(2.54±0.43)vs.(2.20±0.29),(2.16±0.32)]and CHA2DS2-VASc [(3.02±0.89) scores vs.(2.64±0.77)scores vs.(2.33±0.48)scores]in aged group and advanced aged group,P<0.01 all;but there were no significant difference between aged group and advanced aged group (P>0.05).There were no signif- icant difference in incidence rates of mild hemorrhage (21.1% vs.14.7% vs.24.6%)and severe hemorrhage (1.8% vs.1.3% vs.1.5%)among middle-aged group,aged group and advanced aged group,P>0.05 all.Conclu-sion:When INR is closely monitored,INR controlled within 1.6-2.5 warfarin anticoagulation is safe and effective for in aged patients with nonvalvular atrial fibrillation.

Objective: To analyze long-term therapeutic effect and safety of warfarin anticoagulant therapy of different intensity on aged patients with nonvalvular atrial fibrillation (NVAF). Methods: According to age, a total of 197 NVAF patients followed up for five years were divided into advanced aged group [n=65，≥80(85±2.09)years], aged group [n=75, 65-79(76.5±2.27) years] and middle-aged group [n=57, ＜65(57.4±2.18)]. All enrolled patients received long-term warfarin anticoagulant therapy, advanced aged group and aged group received low intensity anticoagulation, international normalized ratio (INR) was 1.6~2.5, while middle-aged group received standard intensity anticoagulation and the INR was 2.0~3.0. Thrombus events and incidence rates of hemorrhage etc. over five years were compared among three groups, and the safe dose range of warfarin was explored. Results: During five-year follow-up, no acute cerebral infarction occurred in three groups. The bleeding and other adverse reaction among three groups were no significant difference（P＞0.05). Compared with middle-aged group, there were significant reductions in warfarin dose [(3.29±0.49) mg/d vs. (2.95±0.38) mg/d, (2.85±0.49) mg/d],INR [(2.54±0.43) vs. (2.20±0.29), (2.16±0.32)] and CHA2DS2-VASc [(3.02±0.89) score vs.( 2.64±0.77) score vs.( 2.33±0.48) score]in aged group and advanced aged group, P＜0.01 all; but there were no significant difference between aged group and advanced aged group (P>0.05). There were no significant difference in incidence rates of mild hemorrhage (21.1% vs. 14.7% vs. 24.6%) and severe hemorrhage (1.8% vs. 1.3% vs. 1.5%) among middle-aged group, aged group and advanced aged group, P>0.05 all. Conclusion: When INR is closely monitored, INR controlled within 1.6~2.5, warfarin anticoagulation is safe and effective in aged patients with nonvalvular atrial fibrillation.

Promoter-trap strategy for enriching targeted colonies has been usually used to elevate the gene targeting efficiency in somatic cells. Knocking out Prnp in animals by gene targeting can render them resistant to Prion diseases. We constructed a bovine Prnp promoter-less targeting vector BoPrPneo, then transfected the linearized vector into the bovine fetal fibroblasts BFF through electroporation. After selecting in cell culture medium with 250 microg/mL G418, we obtained 99 drug-resistant cell colonies, 4 of them were positive for targeted events after PCR screening, and the targeted colonies were further confirmed by sequencing and Southern blotting. This suggests that one allele of Prnp has been successfully knocked out in bovine fetal fibroblasts. This research supplies a simple, safe and effective method to targeting bovine Prnp.
Animals ; Cattle ; Electroporation ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; methods ; Prion Diseases ; genetics ; prevention & control ; Prions ; genetics ; Promoter Regions, Genetic ; genetics ; Transfection

Objective To observe the level of imprinting gene p57kip2 and p27kip1 expression in different hydatidiform moles.To investigate the value of combined detection of p57kip2 and p27kip1 in diagnosis and differential diagnosis of hydatidiform moles.Methods We examined the immunohistochemical staining of p57kip2 and p27kip1 in 30 cases of complete hydatidiform moles and 86 cases of partial hydatidiform moles and 30 cases normal placenta,and also analyze the differences and correlation between the two genes of p57kip2 and p27kip1 in two different patterns of hydatidiform moles.Results The rate of expression of p57kip2 and p27kip1 in complete hydatidiform moles was obviously lower than that in partial hydatidiform moles and nor-mal placenta.There were significant differences in the expression of p57kip2 and p27kip1 among complete hy-datidiform moles,partial hydatidiform moles and normal placenta(P<0.05).p57kip2 had positive correlation with p27kip1 in complete hydatidiform moles(r=0.750,P<0.01),meanwhile there was negative correlation between p57kip2 and p27kip1 in partial hydatidiform moles(r= -1.000,P<0.01).Conclusion The differen-tial expression of p57kip2 and p27kip1 in complete hydatidiform moles and partial hydatidiform moles can be of certain value in the differential diagnosis of hydatidiform moles.Meanwhile,the combined methed is useful to the identification and classification of hydatidiform moles.