OBJECTIVE: To investigate the effect of high glucose on the expression of c-met in human kidney fibroblasts in vitro, and to explore the regulation of Radix Astragali. METHODS: A cell culture system of human kidney fibroblasts was developed in vitro. The human kidney fibroblasts were divided into normal control group, high glucose group and mannitol group. Expressions of c-met and transforming growth factor-beta1 (TGF-beta1) mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR) and the expressions of c-met protein were analyzed by Western blot method after 6-, 12-, 24-, 48- and 96-hour culture. The human kidney fibroblasts were also cultured with 10% Radix Astragali containing serum; the expressions of c-met mRNA and protein were detected after 24- and 48-hour culture. RESULTS: Compared with the normal control group, expression of c-met mRNA in the high glucose group was significantly increased after 12-hour culture (P<0.05), arriving at the peak after 24-hour culture (P<0.01). The level of TGF-beta1 mRNA was higher in the high glucose group than that in the normal control group after 24-hour culture (P<0.05), arriving at the peak after 96-hour culture (P<0.01). Forty-eight hours after treating with 10% Radix Astragali containing serum, the levels of c-met mRNA and protein in fibroblasts were increased, and were higher than those in the high glucose group (P<0.01, P<0.05). CONCLUSION: High glucose can induce the expressions of c-met mRNA and protein in earlier period, and then inhibit the expressions. Radix Astragali can up-regulate the expressions of c-met mRNA and protein of human kidney fibroblasts, which may be one of its action mechanisms in delaying the progression of diabetic nephropathy.

Objective To explore the clinical significance of interstitial chemotherapy applied in breast cancer.Methods Sixty breast cancer patients with modified radical mastectomy were divided into treatment group (40 cases) and control group (20 cases) according to whether using fluorouracil implants.The control group was given conventional treatment.In treatment group,distilled water washed the wound after conventional treatment,and fluorouracil implants (0.4 g) was placed below local tumor,between pectoralis major and pectoralis minor,below 0.5 cm axillary vessel.The bone marrow suppression,liver and renal function,gastrointestinal reaction,skin flap necrosis,subcutaeous hydrops,local recurrence were observed in the 2 groups.Results There were no significant differences in the rates of bone marrow suppression,liver and renal function abnormal,skin flap necrosis,subcutaeous hydrops and gastrointestinal reaction between treatment group and control group (P ＞ 0.05).The rate of local recurrence in treatment group was significantly lower than that in control group [0 vs.20.0％ (4/20)],there was statistical difference (P ＜0.05).Conclusion Intraoperative application of fluorouracil implants is safe and effective in modified radical mastectomy of breast cancer.

Objective To examine the expression of hepatocyte growth factor(HGF) receptor (c-met) and investigate the changes in activity of the HGF/c-met system in human kidney fibroblast by high glucose. Methods HGF and c-met mRNA levels in fibroblast induced by high glucose were detected by RT-PCR. C-met protein was examined by Western blotting. At the same time, the expression of TGF-? and c-met in the exogenous HGF and treating with anti-c-met antibody in vitro were measured. Results Extremely rapid induction of HGF and c-met mRNA was observed at the first six hours by high glucose. On the other hand, both c-met mRNA and c-met protein were markedly increased. HGF (50 ng/ml) induced the expression of c-met ( P

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

Adult ; Aneurysm, Dissecting ; complications ; Aortic Aneurysm ; complications ; Humans ; Lower Extremity ; physiopathology ; Male

Objective To solve the key problem on classification and identification of Salvia miltiorrhiza,which was urgently needed to be solved in breeding of S. miltiorrhiza. Methods Sequence-related amplified polymorphism (SRAP) was carried out to analyze the genetic variation of 48 germplasm in S. miltiorrhiza from different sources. Results The data showd that 120 alleles had been found using 15 pairs of primer which had been selected from 100 pairs. Unweighted pair-group method with arithmetical averages (UPGMA) cluster analysis was used to classify the germplasm of S. miltiorrhiza. Two groups could be divided and three subgroups were contained in every group. Conclusion A complex relationship exists between the genetic distance and space distance of differernt S. miltiorrhiza. The genetic diversity in different geographical populations of S. miltiorrhiza in China is rich. The results show that the genetic distance of different germplasm will increase with the stage of domestication.

Objective To develop a questionnaire to measure the patients related knowledge,attitude,and practice about their hospitalization virtue behaviors and test its reliability and validity.Methods Using KAP theory as a conceptual framework,and using experts consultation and investigating medical staff as a tool to develop items of the scale.The reliability and validity of the questionnaire was tested in 200 patients.Results The questionnaire consisted of three dimensions and thirteen items,and three factors were extracted by principal components analysis,which contributed to 58.211％ of the variance.The Cronbach's α was 0.822 for the total questionnaire,and the content validity index was 0.923.Conclusions The questionnaire shows acceptable reliability and validity,and can be applied to related clinical investigation.

OBJECTIVETo compare the therapeutic effects of peripheral facial paralysis in acute stage by different interventions and explore the better treatments of peripheral facial paralysis.

METHODSOne hundred and thirty one cases of Bell's facial paralysis were randomly divided into three groups. In acupuncture group (44 cases), Dicang (ST 4), Jiache (ST 6), Hegu (LI 4), Yangbai (GB 14) and Taiyang (EX-HN 5), etc. were applied; in electroacupuncture group (45 cases), the selection of acupoints and needling method were same as those in acupuncture group, and the electroacupuncture therapy was applied on Dicang (ST 4), Xiaguan (ST 7), Yangbai (GB 14) and Taiyang (EX-HN 5) in acute stage; in medication and acupuncture group (42 cases), Prednisone and Acyclovir were taken by oral administration, Vitamin B1 and Vitamin B12, were applied by intramuscular injection in acute stage, and acupuncture was applied by the way which was same as that in acupuncture group during quiescent and recovery stages. The curative effects were evaluated by House-Brackmann Grading Scale, and the failed rates were observed by follow-up after one and three months.

RESULTSThe cured and markedly effective rates were 79.6% (35/44), 93.4% (42/45) and 78.6% (33/42) respectively in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.05). The cured rates above tympanichord were 54.2% (13/24), 85.2% (23/27) and 48.0% (12/25) in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.01). There was no significant differences of cured rates below tympanichord among three groups (P > 0.05); and the failed rate in electroacupuncture group was much lower than those in acupuncture group and medication and acupuncture group by follow-up after one and three months (all P < 0.01).

CONCLUSIONThe peripheral facial paralysis is effectively treated by electroacupuncture in acute stage, and it suggests that electroacupuncture should be applied early during the acupuncture treatment of peripheral facial paralysis.

Acupuncture Points ; Acupuncture Therapy ; Acyclovir ; administration & dosage ; Adolescent ; Adult ; Aged ; Electroacupuncture ; Facial Paralysis ; drug therapy ; pathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Prednisone ; administration & dosage ; Treatment Outcome ; Young Adult

Objective To explore the effect of excessive fluoride on expression of mRNA and protein of Wnt3a and β-catenin in rats' osteoblasts and its correlation with pathogenic mechanism of fluorosis.Methods Thirty-six healthy SD rats,weighting 100-120 g and according to body mass,were randomly divided into three groups(twelve in each group).The rats of control were fed wich tap water(fluoride ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-fluoride group:5 mg/L,high-fluoride group:50 mg/L) added to the drinking water to establish the chronic fluorosis model.After fed for eight morth,all rats were killed and metaphysic of femoral was collected.Rat dental fluorosis was observed and bone fluorine was detected by ashing-fluorin ion selective electrode method.The content of bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase 5b(TRACP 5b) in rats' serum was detected by enzyme-linked immunosorbent assay (ELISA).The morphologic changes of the bone were observed by microscopy.The expression of mRNA and protein of Wnt3a and β-catenin in osteoblasts of rats was analyzed with gray scale by hybridization in situ and immunohistochemistry methods,respectively.Results Detection rate of dental fluorosis,fluoride contents of urine and bone were significantly increased [control group:0.0％,(1.26 + 0.17)mg/L,(305.58 ± 91.26)mg/kg; low-fluoride group:66.7％,(2.06 ± 0.64)mg/L,(632.33 ±123.21)mg/kg; high-fluoride group:91.7％,(7.69 ± 1.96)mg/L,(1088.75 ± 156.16) mg/kg] in the rats treated with fluoride,the difference between groups was statistically significant(χ2 =21.6; F =36.57,467.02; all P ＜0.05).The contents of BALP and TRACP-5b in rats' serum were significantly different between groups(F =89.57,7.68; all P ＜ 0.05).Compared with control group[(16.24 + 1.57)U/L],the contents of BALP in rats' serum of the low-fluoride and high-fluoride groups[(31.47 ± 5.30) and (54.61 ± 2.27)U/L] were increased gradually(all P ＜0.05).Compared with the low-fluoride group,the value in the high-fluoride group decreased significantly (P ＜ 0.05).The contents of TRACP-5b in rats' serum of low-fluoride group[(3.45 ± 1.85)U/L] were elevated significantly(all P ＜ 0.05) compared with the control group[(1.26 ± 0.23)U/L] and the high-fluoride group[(2.74 ± 1.85)U/L].The bone cortices were thickened and the bone trabecula was broadened,arranged closely together in chronic fluorosis rats with significant difference compared with the control group.In the low-fluoride and high-fluoride groups,the expression levels of Wnt3a and β-catenin mRNA (low-fluoride group:132.87 ± 5.72 and 132.57 ± 9.56; highfluoride group:135.60 ± 6.64 and 137.87 ± 9.16) were markedly elevated with significant difference,respectively (F =12.47,5.96; all P ＜ 0.05) compared with those in control groups(119.86 ± 5.04 and 120.58 ± 7.84) by hybridization in situ(P ＜ 0.05),but there was no statistical significance (P ＞ 0.05) of the level of Wnt3a and β-catenin mRNA between low-fluoride and high-fluoride groups.In the low-fluoride and high-fluoride groups,the protein expression of Wnt3a and β-catenin (low-fluoride group:137.50 ± 4.32 and 140.85 + 3.54; high-fluoride group:142.65 ± 11.84 and 152.52 ± 4.64) were markedly elevated with significant difference,respectively (F =10.07,53.82; all P ＜ 0.05) compared with those in control group (124.01 ± 2.63 and 126.75 ± 4.65) by immunohistochemistry(all P＜ 0.05),Wnt3a protein production in the low-fluoride group was increased without statistical significance compared with the high-fluoride group (P ＞ 0.05).But the protein production of β-catenin in the lowfluoride group was elevated with significant difference compared with the high-fluoride group(P ＜ 0.05).The mRNA and protein production of Wnt3a were positively correlated with the mRNA and protein production of β-catenin (r =0.731,0.658; all P ＜ 0.05).Conclusions Rat bone tissue lesions caused by excessive fluoride may be associated with an increased expression of Wnt3a and β-catenin mRNA and protein in osteoblasts.In chronic fluorosis,fluoride stimulates the overexpression of Wnt3a and β-catenin in the Wnt signal transduction pathway,enhances bone osteogenesis and causes skeletal fluorosis.

Aim To study the effect of Baoxinbao film on endothelin(ET) and nitric oxide(NO) secretion in patients with stable angina pectoris(SAP).Methods 76 patients with SAP were randomly divided into two groups, with 40 cases in the baoxinbao group plastered with baoxinbao film and 36 cases in the isosorbide dinitrate group receiving isosorbide dinitrate. The levels of plasma ET and NO before and after treatment were observed. Results The concentrations of plasma ET were increased and plasma NO reduced significantly in the SAP patients respectively, as compared with those in the control group(all P