Objective To investigate the relationship between plasminogen activator inhibitor-1(PAI-1) activity and PAI-1 promoter 4G/5G polymorphism in patients with type 2 diabetes from the coastal areas of Shandong province.Methods The 4G/5G allele polymorphism in the PAI1 gene promoter region were tested by allele specific PCR in 116 type 2 diabetes and 40 normal controls.The activity of plasma PAI-1 was assayed by chromogenic substrate method.Results The plasma PAI-1 activity in patients was higher than that in controls(P

Objective To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.Methods Male Wistar rats were randomly divided into sham operation group (n =8),sham operation + apocynin treatment group (n =8),UUO operation group (n =8) and UUO operation+apocynin treatment group (n =8).Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery.Rats were sacrificed at 7th day.ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT),and the level of 8-iso-prostaglandin F2alpha (8-isoPGF2α) in renal tissue.Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4,α-smooth muscle actin(α-SMA),collagen Ⅰ (COL-Ⅰ) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).Results UUO rats with vehicle displayed increased oxidative stress,as measured by renal tissue 8-iso-PGF2α,accompanied with increased renal expression of NADPH oxidases (NOX2,1.5-fold and NOX4,1.7-fold,respectively),compared with sham-operated rats (P ＜0.05).Furthermore,vehicle treated UUO rats showed increased renal COL-Ⅰ and α-SMA levels,compared with sham-operated rats (P ＜ 0.05).ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P ＜ 0.05).Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7％) and NOX4 (-31.0％) in UUO rats,respectively,compared with vehicle treated rats (P ＜ 0.05).And significant decrease of COL-Ⅰ (-26.4％) and α-SMA expression (-80.0％) were also observed (P ＜ 0.05).The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P ＜ 0.01).Despite the pronounced dysregulation of pro-oxidative NOXs family,no compensatory increase of antioxidative enzyme activities occurred.Conclusion The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation,and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.

Objective To establish a cosmetic medicine bio-psycho-social model and its theory frame and a new clinical work system. Methods The clinical observation and clustering analysis were used in this study, which based on the modern medicine patterns to carry on the logical proof, theory derivation and systematic design. Results The bio-psycho-social model of cosmetic me dicine with combination of the theory and practice was established, including its concept, theory constitution and clinical work system. Conclusion The bio-psycho-social model of cosmetic medicine can most scientifically and comprehensively explain this discipline rule and clinical work, and it will bring the profound influence on the development of the theory and practice in cosmetic medicine.

Objective To observe the clinical efficacy of acupuncture-moxibustion plus moving cupping on the back in treating sub-health in the undergraduates. Method According to the theory of preventive treatment in the Chinese medicine, 30 undergraduates with sub-health were treated by acupuncture at Baihui (GV20), Guanyuan (CV4), and bilateral Zusanli (ST36), plus moving cupping to the Governor Vessel, totally for 20 d. The Self-rating Sub-health Scale (SRSHS) was adopted for evaluation before and after intervention, for observing the clinical efficacy. Result Of the 30 subjects, 14 were recovered, 14 showed effective, 2 failed, and the total effective rate was 93.3%. The SRSHS score dropped significantly after intervention (P＜0.001). The psychological factor responded most significantly to the intervention, followed by the physical factor, while the social function factor didn’t show a significant change. Conclusion Acupuncture-moxibustion plus moving cupping is significantly effective in improving and treating the sub-health state in the undergraduates.

OBJECTlVE To develop an LC-MS/MS method for simultaneous determination of pro-damine ( PDM) and its metabolite 2,4-dinitro-N3-propyl-6-trifluoromethyl-1,3-benzenediamine ( DTB) in rat plasma in order to study toxicokinetics of PDM in rats. METHODS SD male rats were administered a single dose of PDM ( ig: 100 and 1000 mg·kg-1; iv: 100 mg·kg-1 ) . LC-MS/MS method was used to determine PDM and DTB in rat plasma. Toxicokinetic parameters were fitted using DAS Ver2. 1. 1. RESULTS After ig administration of PDM 100 mg·kg-1 , the parameters of PDM and DTB were as fol-lows:AUC(0-t) was 2715±102 and (6845±316)μg·h·L-1, t1/2z was 9.0±1.4 and (7.1±1.3)h, Tmax was 7.0± 1.6 and (7.0±0.0)h, cmax was 146±51 and (473±103)μg·L-1. After ig administration of PDM 1000 mg·kg-1, the parameters of PDM and DTB were as follows:AUC(0-t) was 3401±242 and (10364± 573)μg·h·L-1, t1/2z was 8.8±2.1 and (6.0±1.8)h, Tmax was (7.0±1.6)h, cmax was 175±56 and (586± 152)μg·L-1 . The absolute bioavailability of PDM was 44.9%( 100 mg·kg-1 ) and 17.1%( 1000 mg·kg-1 ) . CONCLUSlON This method is suitable for the analysis of PDM and DTB in rat plasma. There is evidence that PDM and DTB display nonlinear toxicokinetic characteristics in the studied dose range.

Objective To make students at different levels not only learn theoretical knowledge of acupuncture-moxibustion manipulation skills but also achieve full operating training, strengthen the combination of theory and practice and improve the teaching quality of clinical disciplines by applyingmultimedia-subject participationteaching method to acupuncture-moxibustion manipulation techniques in the experimental teaching of acupuncture and moxibustion. Method The students were randomized, according to grade, into experiment (n=114) and control (n=115) groups. The teaching content was the same in the two groups. The experiment group received multimedia-subject participationteaching method. After the multimedia teaching, the students did an independent demonstration and made a self-evaluation, the teachers made a comment and the students did independent practice again in groups. The control group received conventional theory teaching and did practice under demonstration in the experimental lesson. The examination and questionnaire were used as assessment indicators to evaluate the effect ofmultimedia-subject participationteaching method applied to acupuncture-moxibustion technique teaching. Result Statistical analysis was made using the SPSS11.5 statistical software package. The independent samples t-test is used for a between-groups comparison. The statistical result showed that satisfaction was 100%in the experiment group and 75.7%in the control group;there was a statistically significant difference between the two groups (P<0.05). Conclusion Multimedia-subject participationteaching method is an innovation in acupuncture-moxibustion technique teaching and helps the students to better master acupuncture-moxibustion manipulation skills. Mastery of theoretical knowledge before real practice is the key to proficient manipulation. The combination of the two in classroom teaching yields twice the result with half the effort. Carrying out multimedia-subject participation teaching method will help to further deepen the teaching reform and to improve the level and quality of“Acupuncture and Moxibustion”teaching.

Objective To investigate the apoptosis of bone marrow mesenchymal stem cells(BMSCs)from systemic lupus erythematosus(SLE)patients and the expression of apoptotic molecules.Methods BMSCs were isolated from bone marrow of SLE patients and normal controls by density eentrifugation and adhesive culture in vitro.The apoptosis of BMSCs were evaluated by TUNEL assay.The expressions of Fas.Bcl-2 and the activity of Caspase 8 were detected by flow cytometry.Real-time PCR technique was used to determine the gene expressions of Fas,Bcl-2,Bax,Bcl-w,Caspase 8 and Apaf-1.Meanwhile,cytochrome C was detected by immunocytochemistry.Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results The percentage of apoptotic BMSCs increased in SLE patients compared with healthy donors[(64±10)%vs [14±9)%,U=0,P<0.05 ].The expression of Bcl-2 in BMSCs of SLE patients was lower than the normal controls at protein leve[(11±9)%vs(56±18)%,U=0,P<0.05 ],and mRNA level(0.2±0.2vs 2.4±0.7,U=24,P<0.05).More cytochrome C positive pellets in the cytosolic fraction could be detected in BMSCs from SLE patients compared with healthy controls [(56±21)%vs (16±16)%,U=1,P<0.05].The activity of Caspase 8 was enhanced[(49±14)%vs(16±12)%,U=0.P<0.05 ],although with no significant difierence at mRNA Ievel.Both groups expressed Fas but with no significant difference (U=19,P>0.05).Conclusion BMSCs from SLE patients undergo more apoptosis,the mechanisms may be associated with the down regulation of Bcl-2,up-regulation of Cytoehrome C in cytoplasm and the activation of Caspase 8,which directs the intrinsic and extrinsic apoptosis pathways.

Objective To determine the molecular mechanism of berberine (BBR) inhibiting the metastasis and invasion of nasopharyngeal carcinoma 5-8F cells, and identify whether berberine suppresses the tumor-invasive action through inhibiting Ezrin or phosphate-Ezrin. Methods The non-cytotoxic concentration of berberine was detected by MTT assay. Filopodia formation of 5-8F cells was observed by electron microscope. The invasion and motility of 5-8F cells with berberine treatment were measured with Trans-well assay. Western blot was used to investigate the Ezrin and phos-Ezrin expression in 5-8F cells treated by berberine. pcDNA3.1-Ezrin and pcDNA3.1-Ezrin M were transfected into 6-10B cells. The inhibitory effect of berberine on the motility and invasion of 6-10B-pcDNA3.1-Ezrin and 6-10B-pcDNA3.1-Ezrin M was detected, respectively. Results Berberine non-cytotoxic concentration was 0-40 μmol/L. After being treated by berberine, filopodia of 5-8F cells obviously reduced, and the permeating artificial basement membrane cells largely decreased in both time- and concentration-dependent manner. There was significant difference compared with the control group (P<0.05). Berberine suppressed the phos-Ezrin expression of 5-8F cells in both time- and concentration-dependent manner (P<0.05), but the effect of berberine was weaker on 6-10B-pcDNA3.1-Ezrin M than on 6-10B-pcDNA3.1-Ezrin. Conclusion Berberine inhibits nasopharyngeal carcinoma cell invasion through inhibiting phos-Ezrin expression and filopodia formation.

Objective To explore the role of intracellular reactive oxygen species (ROS) in the senescence of bone marrow mesenchymal stem cells(BMSCs)in patients with systemic lupus erythematosus(SLE) and the underlying mechanisms that controls the intracellular ROS levels in vitro. Methods Human bone marrow aspirates were collected from iliac of eight donors and eight SLE patients and cultured in vitro.Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was measured by fluorescence microscope. BMSCs were monitored using the senescence associated β-galacto-sidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expression of FoxO3, phospho-FoxO3 (p-FoxO3),AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis. Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results There were no differences in morphology and nuclear size[(31.4±4.5) vs (28.2±4.8) μm, P=0.628] of BMSCs between SLE patients and normal controls.The percentage of SA β-gal positive BMSCs from SLE patients was higher than that from healthy controls [(31.8±9.0)% vs (12.4±0.7)%, P＜0.05]. Intracellular ROS levels of BMSCs from SLE patients increased more significantly than healthy donors in vitro (34600±9600 vs 17 958±5400, P＜0.05). No significant differences in the expression of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a similar upward trend. The expression of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels.Conclusion These data suggest that BMSCs from SLE patients aged more quickly, with high SA β-gal activity and up-regulation of intracellular ROS, which is associated with up-regulation of p-FoxO3 and pAKT at protein levels. These results indicate that bone marrow mesenchymal cell senescence may be associated with the pathogcnesis of SLE by maintaining the lifespan of BMSCs.