Objective:To explore bromodomain and extra-terminal (BET) inhibition in the regulation of vascular endothelial cells activation and early atherosclerosis formation and its potential molecular mechanisms.Methods:1.Human umbilical vein endothelial cells (HUVEC) and mouse heart endothelial cells (MHEC) were isolated,and tumor necrosis factor α (TNFα) was used to activate in flammatory genes transcription in the presence or absence of JQ1,a specific BET inhibitor.The groups are as follows:(1)Normal control group;(2) TNFα(25 ng/mL)group;(3) TNFα+JQ 1 group.The gene mRNA and protein expression of inflammatory cytokines were measured by both real-time PCR and flow cytometry (FCM).2.LDL receptor-deficient (LDLR-/-) mice were randomly divided into 2 groups:JQ1 group (n=8,JQlintraperitoneal,50 mg/kg,daily) and control group (n=8,DMSO,daily).After 8 weeks feeding with high cholesterol diet,vascular cell adhesion molecule-1 (VCAM-1) expression in aortic arch was measured by immunohistochemistry.The activity of nuclear factor kappa B (NF-κB) signaling was monitored by 5XκB luciferase reporter assay in HEK293.Results:TNFα dramatically induced the mRNA and protein expression of inflammatory genes and JQ 1 significantly downregulated the induction of them (E-selectin,P-selectin,VCAM-1,IL-8)(P＜0.01).Immunohistochemistry detection indicated that JQ1 significantly downregulated the expression of VCAM-1 in aortic arch induced by 8 weeks high cholesterol diet feeding comparing to control group.In addition,BET bromodomain inhibition downregulated TNFα upregulated NF-κB transcriptional activity (P＜0.01).Conclusions:Our study demonstrated that BET bromodomain was involved in NF-κB mediated inflammatory genes expression;inhibition of BET bromodomain suppressed vascular endothelial activation in vitro,and attenuated early atherogenesis in vivo.

Objective To evaluate the effect of CT temporal subtraction(TS) for detection of lung nodules.Methods 80 cases of CT images (current and previous CT images) and corresponding CT TS images were presented, which included 30 cases with nodules(75 nodules) and 50 controls.4 observers, including 2 radiologists and 2 residents, indicated their confidence level regarding the presence of a new emerging or larger or solid component than before,which first used standard CT images, then with the addition of CT TS images.Receiver operating characteristic(ROC) curve analysis was used to evaluate the observers'' performance.Results The mean value of the area under the ROC curve (AUC) were 0.860 and 0.925 for four observers without and with TS images,respectively(P<0.01).Average sensitivity for detection of lung nodules was improved from 77.3%(58/75) to 89.3%(67/75) by using CT TS images.Conclusion The use of CT TS images can significantly improve radiologists'' performance for detection of lung nodules,especially when a small size solid or subsolid nodule presents near the pulmonary hilum.CT TS is more helpful for residents.

Objective:To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. Results:After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (allP<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Conclusion: Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.

Objective To investigate the predictive value of the whole nodule size and solid component size of lung adenocarcinoma manifesting as subsolid nodule(SSN) in three different dimensions for pathologic grade.Methods We evaluated retrospectively preoperative chest HRCT data of 125 patients with 127 SSNs surgically resected and pathologically conformed lung adenocarcinomas.All specimens were divided into two groups: a total of 69 SSNs in group A, including 22 AIS and 47 MIA;a total of 58 SSNs in group B, only including IAC.Computer aided diagnosis software were used to measure the one dimension maximum diameter of solid component with lung window setting(1D-SCLW),two dimension maximum diameter of solid component with lung window setting(2D-SCLW),one dimension maximum diameter of solid component with mediastinal window setting(1D-SCMW),two dimension maximum diameter of solid component with mediastinal window setting(2D-SCMW),one dimension maximum diameter of whole nodule with lung window setting (1D-WNLW), two dimension maximum diameter of whole nodule with lung window setting (2D-WNLW), and volume of solid component with threshold of-300 HU (SCT) of all SSNs.Results 1D-SCLW, 2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW,2D-WNLW and SCT of the group B were significantly larger than those of the group A(P=0.000).ROC analyses indicated that the diagnostic efficiency of SCT for the pathologic grade was the highest among 7 CT features(AUC=0.887, sensitivity:81%,specificity:93%);The cut-off values of 1D-SCLW,2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW, 2D-WNLW and SCT were 17.50 mm,14.75 mm,9.50 mm,7.75 mm,0.50 mm,1.25 mm and 139.00 mm3.Multiple Logistic regression analysis revealed that SCT was the independent predictor of pathologic grade(OR=4.978,95%CI=1.430-17.331,P=0.012).SCT of 139.00 mm3 or greater was a significant indicator of IAC.Conclusion Among the whole nodule size and solid component size of SSN in three different dimensions on preoperative HRCT, SCT is found to be the independent predictor of pathologic grade, which may provide reference for surgery.

Objective To evaluate the performance of bone suppression images on the detection of lung nodules in comparison with the radiologists'reading results.Methods There were 141 standard posteroanterior digital chest radiographs,which included 95 patients with a solitary nodule and 46 controls.In this observational study,4 observers,including 2 radiologists and 2 residents,in-dicated their confidence level regarding the presence of a nodule for each lung,first by use of standard images,then with the addition of bone suppression images.Receiver operating characteristic (ROC)curve analysis was used to evaluate the observers'performance. Results Average nodule size was (1.9±1)cm (range from 0.9 cm to 2.9 cm).The mean value of the area under the ROC curve (AUC)was significantly improved from 0.844 with use of standard images alone to 0.873 with use of bone suppression images (P<0.01).Conclusion The use of bone suppression images can improve radiologists'performance on detection of lung nodules on chest radiographs.

· AIM: This study was designed to evaluate the effectiveness of anterior lens capsule inclusion in combined trabeculectomy and cataract surgery in preventing scar formation of the filtering bleb in rabbit model.· METHODS: Twerty-Four eyes (12 rabbits) with glaucoma model were studied, anterior lens capsule inclusion in trabeculectomy with the small-incision cataract surgery were performed on all right eyes (experimental group) and no implants were applicator in trabeculectomy with the small-incision cataract surgery on all left eyes (control group). These operated eyes were followed up from day 1 to 6 months postoperatively. Intraocular pressure (IOP) was measured and filtering blebs were observed after surgery. Other main outcome measures: cornea、conjunctiva、formation of the anterior chamber, anterior chamber depth、inflammatory reaction、achievement ratio of operation and complications were analyzed. On week 1, 2, 4, 12 and 24 after surgery the animal were killed in batch. Tissue was harvested from the bleb area and was made pathological section. HE staining、light microscope and micro photo analysis technique were applied to observe the cytological and histopathologic characteristics of the filtering tunnels.· RESULTS: There were significant differences between the two groups on IOP (1, 2, 4 weeks)、filtration bleb, achievement ratio of operation and complications. In experimental group, at the first month postoperatively, anterior lens capsule absorption started with inflammatory characteristics. The peak of inflammatory reaction occurred 1 week after operation and all the cells in the filtrating tunnel disappeared 6month after surgery. The fibroblast proliferation in control group occurred at I week and the filtrating tunnel closed with angiogenesis at 1 month after surgery. Fibroblast proliferation started 1week after surgery with no statistical difference during the time course (P ＞0.05). Significant statistical differences were observed by comparing the fibro blasts numbers per unit area in the filtrating tunnel in experimental group and those in control groups (P＜0.05).· CONCLUSION: Anterior lens capsule was totally absorbed at 6 months postoperatively. Anterior lens capsule inclusion in trabeculectomy with cataract surgery can possibly control intraocular pressure effectively, long-term sustainability of functional filtration bleb, inhibition of the proliferation of fibroblasts and opening of the filtrating pathway in the experimental animal models were satisfied. Compared to the control group, anterior lens capsule application has less complication.

Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06％ ± 6.37％ vs.100.00％ ± 10.42％ and 108.70％ ± 9.90％ at 24 hours,42.93％ ± 5.93％ vs.100.00％ ± 6.24％ and 168.00％ ± 2.98％at 48 hours,all P ＜ 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P ＜ 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P＜ 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P＜ 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P ＜ 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P ＜ 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30％ vs.0.421％ and 2.22％ at 24 hours,4.06％ vs.0.577％ and 0.691％ at 48 hours,all P＜ 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.

Objective To investigate the predictive value of whole nodule size and solid component size of pulmonary subsolid nodules (SSNs)with different window setting on preoperative HRCT for pathologic grade in lung adenocarcinoma.Methods We retrospectively evaluated preoperative chest HRCT and pathological data of 125 patients with 127 surgically resected lung adenocarcinoma manifesting as SSNs.All specimens were divided into two groups:a total of 69 SSNs in group A,including 22 adenocarcinomas in situ (AIS) and 47 minimally invasive adenocarcinoma (MIA);a total of 58 SSNs in group B,including invasive adenocarcinoma (IAC).Observer 1 used computer aided diagnosis software to measure the volume of whole nodule with lung window setting (WNLW),volume of solid component with lung window setting (SCLW),volume of solid component with mediastinal window setting (SCMW) and volume of solid component with threshold of-300 HU(SCT) of all SSNs.Observer 2 randomly selected 50 SSNs and repeated all the measurements.The interobserver agreement regarding quantitative measurements were evaluated by using intraclass correlation coefficient(ICC).The differences of all quantitative features between two groups were evaluated by Mann-Whitney U test.All the quantitative features were evaluated by using univariate logistic regression analysis,significant quantitative features identified by univariate logistic regression analysis were included in the multivariate logistic regression and independent predictors of pathological grade were obtained.Receiver operating characteristic analysis was conducted for the independent predictive factors that exhibited statistically significant differences in the multivariate logistic regression.Results The interobserver agreement regarding quantitative features were excellent (ICC＞ 0.75).The WNLW,SCLW,SCMW and SCT of group B were significantly larger than those of group A (P＜ 0.001).The univariate logistic regression analysis indicated that WNLW,SCLW,SCMW and SCT were significant (P＜0.001),the multivariate logistic regression analysis indicated that SCT was the independent predictive factor (OR=1.013,95％CI:1.006—1.020,P＜0.001).When SCT larger than 139.00 mm3,SSN was significantly associated with IACs (AUC=0.887,sensitivity=81％,specificity=93％).Conclusion SCT of SSNs on preoperative HRCT can be used to distinguish between AIS-MIA and IAC,which may provide information for choice of operation.

OBJECTIVETo investigate the adverse effects of acupuncture on the prognosis, and effectiveness of acupuncture combined with far infrared ray in the patient of acute Bell's facial paralysis within 48 h.

METHODSClinically randomized controlled trial was used, and the patients were divided into 3 groups: group A (early acupuncture group), group B (acupuncture combined with far infrared ray) and group C (acupuncture after 7 days). The facial nerve functional classification at the attack, 7 days after the attack and after treatment, the clinically cured rate of following-up of 6 months, and the average cured time, the cured time of complete facial paralysis were observed in the 3 groups.

RESULTSThere were no significant differences among the 3 groups in the facial nerve functional classification 7 days after the attack, the clinically cured rate of following-up of 6 months and the average cured time (P > 0.05), but the cured time of complete facial paralysis in the group A and the group B were shorter than that in the group C (P < 0.05).

CONCLUSIONThe patient of acute Bell's facial paralysis can be treated with acupuncture and moxibustion, and traditional moxibustion can be replaced by far infrared way.

Acupuncture Therapy ; Bell Palsy ; therapy ; Facial Paralysis ; therapy ; Humans ; Moxibustion ; Treatment Outcome

OBJECTIVETo study if Scleraxis, a basic helix-loop-helix type transcription factor, could be expressed in human periodontal ligament cells (hPDLC), bone marrow cells (hBMSC) and gingival fibroblasts (hGF), and to investigate if Scleraxis was involved in hPDLC differentiation.

METHODShPDLC, hBMSC and hGF were cultured. Expression of Scleraxis in hPDLC from different passages and in hBMSC and hGF was analyzed by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSScleraxis expression in hPDLC, hBMSC and hGF were significantly different (P < 0.05). The A values of Scleraxis/beta-actin in these kinds of cells were 0.877 +/- 0.024, 0.438 +/- 0.031, 0.313 +/- 0.083, respectively. The expression of Scleraxis was the highest in hPDLC and lowest in hGF. Scleraxis expression of hPDLC decreased with increase of passages in culture.

CONCLUSIONSScleraxis was expressed in hPDLC, hBMSC and hGF in vitro, and may play an important role in differentiation of hPDLC.

Adolescent ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Fibroblasts ; metabolism ; Gingiva ; cytology ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Periodontal Ligament ; cytology ; metabolism ; Young Adult