Objective To investigate the effectsof quercetin on human umbilical vein endothelial cell (HUVEC)-12 undergoing endothelial-to-mesenchymal transition (EndMT) induced by TGF-β1; To discuss its mechanism of action. Methods Cell activity of intervening by quercetin with different concentrations and TGF-β1 for 72 h was detected by CCK-8 method;RT-PCR and immunofluorescence staining were used to detect the transition of endothelial and stromal markers; Western blot was used to detect the signal transduction pathway; RT-PCR was performed to detect the transcription factors that play crucial roles in the process of transformation. Results The results of RT-PCR and immunofluorescence double staining showed that TGF-β1 (10 ng/mL) stimulated HUVEC-12 cells for 72 h to induce fibroblast phenotype, showing more interstitial markers and less endothelial markers;Western blot and RT-PCR results showed that quercetin inhibited the phosphorylation of smad2/3 in a concentration-dependent manner;After TGF-β1 stimulation, the downstream transcription factors EndMT of snail1, twist1, twist2, ZEB1, and ZEB2 significantly increased, while 100 μmol/L quercetin could down-regulate the five downstream transcription factors. Conclusion Quercetin has anti-fibrosis effects through inhibiting HUVEC-12 cells undergoing EndMT.

Objective To investigate the risk factors for vascular cognitive impairment and the effet of cerebral microbleeds (CMBs) on cognitive function in patients with ischemic stroke.Methods The data of patients with ischemic stroke over the age of 50 were collected.The Montreal cognitive assessment (MoCA) scale and Alzheimer's disease assessment scale-cognitive subscale were used to evaluate cognitive function.Hamilton depression scale (HAMD) was used to evaluate the depression status in order to exclude the patients with depression.The patients with ischemic stroke were divided into either a cognitive impairment group or a non-cognitive impairment group according to the scale evaluation results.The demographic and clinical characteristics in both groups were compared,and the multivariate logistic regression analysis was used to look for the independent risk factors for cognitive impairment in patients with ischemic stroke.The Spearman rank correlation method was used to analyze the degree of CBMs,total score of MoCA,and the correlations of all cognitive domains scores.Results A total of 169 patients with ischemic stroke were enrolled in the study.There were 80 patients in the cognitive impairment group and 89 in the non-cognitive impairment group; 34 patients had CMBs and 135 had no CMBs.The age was older (71.99 ±6.01 years vs.64.47 ±6.15 years; t =8.014,P =0.000),years of education were fewer (4.51 ± 1.534 years vs.6.94 ±2.357 years; t =8.023,P =0.000),systolic blood pressure was higher (156.19± 17.53 mm Hg vs.142.04± 16.03 mmHg,1 mmHg=0.133 kPa; t =5.479,P =0.000),scale of white matter lesion was higher (7.33 ± 2.04 vs.4.39 ± 2.17; t =8.951,P =0.000),cerebral infarction volume was larger (7 123.8 ± 1 587.1 mm3vs.5 628.4 ± 1 017.8 mm3;t =7.201 ; P =0.000),proportion of the patients with history of previous stroke or transient ischemic attack was higher (46.2％ vs.28.1％;x2 =5.982; P=0.014),and number of CBMs was larger (x2 =17.565; P=0.000) in the cognitive impairment group.Multivariate logistic regression analysis showed that the age (odds ratio [OR] 1.115,95％ confidence interval [CI] 1.013-1.227; P =0.026),years of education (OR 0.490,95％ CI0.325-0.793; P=0.001),systolic blood pressure (OR 1.048,95％ CI 1.014-1.083; P =0.005),scale of white matter lesion (OR 2.044,95％ CI 1.466-2.851; P =0.000),and cerebral infarction volume (OR 2.204,95％ CI 1.386-3.503; P =0.001) were all the independent risk factors for cognitive impairment in patients with ischemic stroke.Compared to the non-CBM group,the age was older (72.06 ± 5.59 years vs.67.01 ±7.15 years; t =4.427; P =0.000),years of education were fewer (3.97 ± 1.381 years vs.6.25 ±2.317 years; t =7.367,P =0.000),systolic blood pressure was higher (155.03 ±20.16 mm Hgvs.147.16 ±17.32 mm Hg; t =2.290,P =0.023),scale of white matter lesion was more higher (7.03 ±2.139 vs.5.47 ±2.591; t =3.247,P =0.001),cerebral infarction volume was larger (6 968.5 ± 1 507.4 mm3 vs.6 177.0 ±1 477.1 mm3; t =2.735,P =0.007),and proportions of hypertension (82.4％ vs.41.5％ ;x2 =18.149,P =0.000),hyperlipidemia (88.2％ vs.39.3 ％ ;x2 =26.067,P =0.000),history of previous stroke or transient ischemic attack (70.6％ vs.28.1％ ;x2 =21.061,P =0.000) and coronary heart disease (94.1％ vs.45.2％ ;x2 =26.278,P=0.000) were higher in the CBM group.The MoCA total score (M[Q1 ～ Q3]; 24 [24 ～25]vs.28 [27 ～ 28] ; Z =-7.092,P =0.000) as well as the scores of attention (6 [5 ～ 6] vs.6 [6 ～ 6] ; Z =-2.502,P =0.012),abstraction (2[1 ～2] vs.2[2 ～2] ; Z =-2.382,P =0.017) and visuoexecutive (2[1 ～2] vs.4[4 ～5]; Z=-7.321,P=0.000) in the CMB group were significantly lower than those in the nonCBM group.The Spearman rank correlation analysis showed that the CMB grade was negatively associated with the MoCA total score (rs =-0.879,P =0.000) as well as the scores of visuoexecutive (rs =-0.895,P =0.000),attention (rs =-0.337,P =0.005),and abstraction (rs =-0.333,P=0.006).Conclusions The age,years of education,systolic blood pressure,degree of white matter damage,and cerebral infarction volume are the risk factors for vascular cognitive impairment.The visuospatial executive dysfunction,attention and abstract thinking decline significantly in ischemic stroke patients with CBMs.CMBs and their numbers are closely associated with cognitive impairment.The more the CMB numbers are,the more obvious the cognitive impairment will be.

OBJECTIVETo explore a safe and effective scheme of analgesic for labor.

METHODSEighty-four primiparas without contraindication of vaginal delivery were divided into an observation group and a control group, 42 cases in each one. In the observation group, the acupoint injection was given at Zusanli (ST 36) combined with electroacupuncture (EA) at Hegu (LI 4) and Sanyinjiao (SP 6) till the cervical opening at the end of first stage labor. In the control group, the routine respiratory instruction was applied. In 5 min, 10 min and 60 min of acupuncture (the same time points in the control group) as well as at the end of the first and second stage labor, the analgesic effect was assessed for the primiparas of the two groups. The labor stages, adverse reactions, postpartum hemorrhage, postpartum urine retention, newborn asphyxia rate and usage rate of oxytocin were compared between the two groups.

RESULTSIn the observation group, in 5 min, 10 min and 60 min of acupuncture as well as at the end of the first and second stages, the visual analogue scale (VAS) was lower apparently as compared with the control group at the corresponding time points (all P < 0.05). The differences in the time limit in the active period, the second and third stages were not significant between the two groups (all P > 0.05). The incidence of adverse reactions and the usage rate of oxytocin were lower than those in the control group [2.4% (1/42) vs 31.0% (13/42); 2.4% (1/42) vs 23.8% (10/42), both P < 0.05]. The differences in postpartum hemorrhage, postpartum urine retention and newborn asphyxia rate were not significant between the two groups (all P > 0.05).

CONCLUSIONThe combination of acupoint injection and EA is the effective analgesic scheme for labor. This scheme effectively alleviates labor pain and has no maternal and child complications.

Acupuncture Analgesia ; Acupuncture Points ; Adult ; Analgesics ; administration & dosage ; Combined Modality Therapy ; Electroacupuncture ; Female ; Humans ; Labor Pain ; drug therapy ; therapy ; Labor, Obstetric ; Pregnancy ; Young Adult

Objective:To explore the effect of autoimmune regulator to TLRs expressions on peripheral antigen presenting cells(APC).Methods:①pEGFPC3-AIRE plasmid was transfected with liposome.②Confocal microscopy was used to observe the effect of transfection.③RT-PCR assay was used to detect the expressions of AIRE and TLR1-9 in RAW264.7cells at 36,48,72,96 h after transfection.Results:①The plasmid was transfected into RAW264.7 cells successfully,and the efficiency of transfection was 60~70%.②AIRE transfected RAW264.7 cells were achieved,and the best time was 72 h.③At 72 h after transfection,the expressions of TLR1,4,5,9 increased,and TLR3,7,8 reduced.The expression of TLR2,6 increased at 96 h.Conclusion:AIRE may regulate the immune response by control TLR expression in APC.It maintain the effective response to pathogen and tolerance state to self tissues through the effects to different TLRs.

Objective To evaluate four tumor markers of insulin-like growth factor 1((IGF-1), CEA, cytokeratin fragment antigen 21-1 (CYFRA21-1), neuron-specific enolase (NSE) for the diagnosis and prediction of treatment response in human lung cancer. Methods Serum samples were taken from three groups: 91 patients with lung cancer, 30 healthy adults and 15 patients with benign pulmonary diseases. Serum IGF-1 was assayed by radioimmunoassay and CEA, CYFRA21-1, and NSE by electrochemiluminescence immunoassay. The differences among the three groups were determined by Kruskal-Wallis one-way analysis of variance (ANOVA) and with Mann-Whitney rank-sum test. Diagnostic efficacy was evaluated by ROC curves. Results The four serum tumor marker levels were significantly higher in lung cancer group, as compared with the benign and the healthy (IGF-1:χ2=26.95,P<0.001, CEA:χ2=49.11,P<0.001; CYFRA21-1:χ2=40.63,P<0.001; NSE:χ2=14.76;P<0.001). The diagnostic sensitivities of IGF-1, CEA, CYFRA21-1 and NSE was 75.6% (34/45), 53.3% (24/45), 66.7% (30/45) and 42.2% (19/45) respectively for lung cancer. The diagnostic sensitivity of IGF-1 combined with CYFRA21-1 was 95.5 %( 43/45) and that of IGF-1 combined with CEA and CYFRA21-1was 97.8%(44/45). Only IGF-1 and CYFRA21-1 showed significant changes before and after treatment (IGF-1: χ2=5.99,P=0.014; CYFRA21-1:χ2=4.99, P=0.025) in cancer group. Conclusions Serum IGF-1, CEA, CYFRA21-1 and NSE are all valuable for lung cancer diagnosis and the combination of those parameters can enhance the diagnostic efficiency. Serum IGF-1 and CYFRA21-1 may also be useful for evaluating the treatment response in lung cancer.

OBJECTIVEThis study aims to evaluate the antimicrobial activity of decapeptide, a novel antimicrobial peptide, against several major cariogenic and periodontopathogenic bacteria in vitro. METHODS In this study, we investigated the antimicrobial activity of decapeptide against Streptococcus mutans (S. mutans), Streptococcus sobrinus, Lactobacillus acidophilus, Streptococcus sanguis, Streptococcus gordonii, Actinomyces viscosus, Actinomyces naeslundii, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Saccharomyces albicans in vitro using the agar diffusion method and broth dilution method. Furthermore, a time-kill kinetic study of decapeptide against S. mutans was performed.

RESULTSThe results showed that decapeptide exhibited antimicrobial activity against various oral bacteria and fungi. The minimum inhibitory concentration (MIC) of main cariogenic bacteria ranged from 62.5 μg · mL(-1) to 125 μg · mL(-1), and the MIC of periodontopathogenic bacteria tested ranged from 250 μg · mL(-1) to 1,000 μg · mL(-1). Among the bacteria tested, decapeptide had a strong inhibitory effect on cariogenic S. mutans. Results of the time-kill kinetic studies showed that decapeptide reduced the viable counts of S. mutans by more than one order of magnitude after 20 min of incubation, and thoroughly killed S. mutans after 30 min. No viable cells could be detected after 24 h of incubation.

CONCLUSIONThis study suggest that decapeptide might have potential clinical application in treating dental caries by killing S. mutans within dental plaque.

Aggregatibacter actinomycetemcomitans ; Anti-Bacterial Agents ; Anti-Infective Agents ; Bacteria ; Dental Caries ; Dental Plaque ; Kinetics ; Microbial Sensitivity Tests ; Mouth ; microbiology ; Porphyromonas gingivalis ; Streptococcus mutans

Objective To investigate the expression and significance of Insulin like growth factor-ⅡmRNA binding protein3 (IMP3) in gastrointestinal stromal tumors (GIST). Methods One hundred and fifteen cases of GIST were subjected to immunohistochemical staining for IMP3 (SP method) and ki67 (iVisionTM method ). The relationship between IMP3 expression with clinicopathologic parameters and Ki67 proliferation indexes were analyzed. In situ hybridization detection of IMP3 mRNA was performed in 20 cases of GIST positive. Results The expression rate of IMP3 was 56.2%(65/115) in GIST positive. A significant correlation can be found between IMP3 expression and tumor of infiltration , coagulative necrosis , metastasis , nuclear atypia , diameter , mitotic count , risk grade of NIH (P < 0.05), while there was no significant correlation between IMP3 and gender, age, primary location, categories of cell type (P>0.05). There was positive correlation of IMP3 expression and Ki67 labeling indexes by Spearman analysis. Positive expression with IMP3 can be observed in 3 cases of recurrence.It may play important role in invasive and development and may be an important factor of prognosis in GIST.

Objective To establish a technical route for the efficient expression and purification of PprI protein from Deinococcus radiodurans R1 by using eukaryotic Pichia pastoris.Methods The encoding sequence of the Deinococcus radiodurans pprI gene was modified according to the preference of Pichia pastoris' codon.Modified pprI gene was fully synthesized with PCR and a 6 × His tag was added at its Nterminal.The PCR products were purified and then cloned into Pichia pastoris expression vector pHBM-905A.After utilizing Cop I and Not I double enzyme digestion and retrievering linear objective fragment,new pprI gene was transformed to the GS115 strain of Pichia pastoris.The obtained Pichia pastoris transformants were induced to express.Culture supernatants were detected by SDS-PAGE,Western blot,and mass spectrometry.A Ni-NTA column was uesd to purify the target protein and the BCA method was used to determine protein concentration.Results The coding sequence of new synthetic Deinococcus radiodurans pprI gene was correct.The purpose protein band of a molecular weight of 43 000 was detected in the culture supernatant of transformed Pichia pastoris strains by SDS-PAGE and Western blot.The mass spectrometry confirmed that it was the Deinococcus radiodurans PprI protein.When the concentration of imidazole was 250 mmol/L,the elution rate of PprI protein was the highest.The purified protein concentration was 0.35 mg/ml measured by BCA method.Conclusions This study has successfully constructed a new pprI gene and the recombinant strain of Pichia pastoris secreting PprI protein,and established a technical route for the efficient expression and purification of PprI protein.