Objective To evaluate the reproducibility of quality intima-media thickness(QIMT) and quality arterial stiffness (QAS) technique in assessment of carotid under different measuring methods.Methods Between December 2012 and January 2014,carotid QIMT and QAS examinations were carried out in 30 health volunteers.QIMT and QAS indicators included IMT in QIMT and distensibility coefficient (DC),compliance coefficient(CC),stiffness index α(α),stiffness index β(β),pulse wave velocity(PWV) in QAS.The measurement employed unilateral/once,bilateral/twice,and unilateral/twice methods.Using intra observer and inter-observer variability,the reproducibility was compared between different QIMT and QAS indicators and measuring methods.Results Extremely high level of intra-observer reproducibility was found for both QIMT and QAS technique (ICC＞0.8).QIMT also showed an excellent inter observer reproducibility (ICC＞0.8).In contrast,the reproducibility of QAS technique varied in different indicators (PWV ＞ β ≈ α ＞ CC ＞ DC) and method ( unilateral/once ＞ bilateral/twice ＞ unilateral/twice).Conclusions QIMT measurement was highly reproducible.Whereas the reproducibility of QAS technique varied in different indicators and methods.Due to low reproducibility,the study result did not support the clinical application of DC indicator and unilateral/once method.

Aim The protective effect and mechanism of TPG on PC12 cells in calcium overloading injury models were studied. Methods Two injury models induced by KCl and NMDA were used to assay the action of TPG in cultured PC12 cells. Results The morphological examination revealed that TPG possessed obvious protective effects on PC12 cells in injury models. MTT and LDH measurement indicated that TPG increased the number of live cells and reduced the extent of cell injury significantly. TPG also lessened the concentration of calcium ion in cytoplasm. Conclusion TPG protected rat PC12 cells against two calcium overloading injuries effectively in vitro. Its actions may deal with anti oxidation, inhibition of NO production and blocking of both types of calcium channel.

Ubiquitination on target proteins is the signal of cellular protein degradation.Ubiquitin ligase E3 is one of the key enzymes in ubiquitination,it recognizes a specific substrate protein and recruits an ubiquitin conjugating enzyme E2,mediating the ubquitin transfer from the E2 to the substrate protein.Ubiquitin ligase E3 can be divided into two subfamilies according to their different structure characters and function mechanisms,the HECT(homologous to E6AP C terminus) family and the RING-finger family.Members of the HECT E3 share the common HECT catalytic domain,which can bind to an E2 and load the ubiquitin on themselves before catalyzing the transfer of ubiquitin to the target proteins.While the RING-finger E3 all contain an similar E2 binding domain and a unique substrate binding part,mediating direct ubiquitin transfer from the E2 to the substrate.The most recent progresses in the stuctural and functional studies of these two E3 famlies were summarized.

Objective To investigate the function of triggering receptor expresses on myeloid cells receptor-1 (TREM-1) in lymphocyte differentiation and regulation of Aspergillus infected immunosuppressed rats.Methods Cyclophosphamide (CTX) was intraperitoneally injected and Fumigatus spore suspension was inhaled by percutaneous tracheostomy to establish the immunosuppressive invasive pulmonary aspergillosis (IPA) rat model.After 24 h, 48 h, 72 h and 96 h inoculation, rats were sacrificed.Lung tissue specimens, bronchoalveolar lavage fluid (BALF) , and plasma samples were collected.Plasma and BALF sTREM-1, plasma T cell-specific transcription factor (T-box expressed in T cells, T-bet) and eomesodermin(Eomes) were detected by ELISA.Biopsy specimens of lung tissue were used for periodic acid-schiff (PAS) staining and culture.Results The mortality rate of immunosuppressed rats after Aspergillus inhalation for 96 h was as high as 54.4％.Biopsy of lung tissue suggested acute inflammatory cell infiltration, interstitial lung congestion, alveolar structural damage, and visible Aspergillus hyphae in alveoli.Compared with normal control group[(110.50 ± 7.70)ng/L], plasma sTREM-1 in study groups were significantly increased [IPA : (146.77 ± 10.41) ng/L;CXT + IPA at 24 h : (226.00 ± 11.88) ng/L;CTX + IPA at 48 h : (200.77 ± 10.63) ng/L;P ＜ 0.05], so were T-bet levels [IPA : (561.17 ± 7.23) μg/L;CXT + IPA at 24 h : (647.00 ± 33.03) μg/L;CTX + IPA at 48 h : (619.23 ± 87.44) μg/L;control group : (340.03 ± 26.32) μg/L;respectively, P ＜0.05].However, plasma Eomes levels in IPA group, CTX + IPA at 24 h and 48 h were significantly lower compared with that in normal controls [IPA : (7.96 ± 0.65) ng/L;CXT + IPA at 24 h : (3.97 ± 0.35) ng/L;CTX + IPA at 48 h : (4.00 ± 0.74) ng/L;control group : (8.38 ± 0.51) ng/L;respectively,P ＜0.001].Compared with those in CTX + IPA vaccination after 24 h and 48 h, plasma sTREM-1 [(106.67 ±7.64)ng/L;(133.27 ± 32.79) ng/L] and T-bet [(299.64±63.07)μg/L;(398.02 ± 109.22) μg/L] in CTX + IPA at 72 h and 96 h inoculation were significantly lower (P ＜ 0.001).While Eomes [(8.38 ± 0.54) ng/L;(8.40 ± 0.70) ng/L] raised significantly higher (P ＜ 0.001).Compared with the control group, sTREM-1 levels in BALF of IPA + CTX 24 h, 48 h, 72 h, and 96 h groups were consistently high (P ＜ 0.05).Pearson correlation analysis showed that sTREM-1 and T-bet had a significant positive correlation (r =0.91, P ＜ 0.001), yet Eomes was negatively correlated with them (r =-0.788, P ＜ 0.001).Conclusions sTREM-1 in rat plasma and BALF appears highly expressed in immune compromised Aspergillus infected rat model.Plasma sTREM-1 is closely correlated with T-bet and Eomes levels, which suggests that TREM-1 may be involved in lymphocytic regulation and differentiation during fungal infection.

Paeoniflorin is the main active ingredient of Chinese herbaceous peony. This study is to investigate the protective effect of paeoniflorin (Pae) on acute brain damage induced by lipopolysaccharide (LPS) in mice. The mice were randomly assigned to the normal control, model control (LPS), as well as groups of paeoniflorin and lipopolysaccharide (Pae + LPS). Then the mice were administered intraperitioneally with normal saline or Pae (10, 30 mg · kg(-1)) once daily for 6 d. One hour after intrapertioneally treatment on the seventh day, each group were injected LPS (5 mg · kg(-1)) to establish the endotoxin lipopolysaccharide inflammation model except the normal group. The mice were sacrificed after 6 h and the brain homogenates were prepared and measured. The malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), succinatedehydrogenase (SDH), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase were dectected by the colorimetric method. The levels of HO-1 and Nrf2 protein in subcellular fractions of brain tissue were detected by Western blot. The results demonstrated that the administration with paeoniflorin reduced the levels of the MDA production; significantly increase the activities of antioxidant enzyme (SOD and GSH-PX). In addition, paeoniflorin could enhance the total antioxidant capacity, decrease the level of H2O2, and increase the activities of SDH, Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase. Furthermore, paeoniflorin can increase the expression of HO-1 and activate the nuclear transfer of Nrf2. Taking together, these findings suggest that paeoniflorin alleviate the acute inflammation in mice brain damage induced by LPS, which is related with its antioxidant effect and improvement of energy metabolism.
Animals ; Energy Metabolism ; drug effects ; Glucosides ; pharmacology ; Heme Oxygenase-1 ; genetics ; Lipopolysaccharides ; pharmacology ; Male ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; pharmacology ; Oxidative Stress ; drug effects ; Sodium-Potassium-Exchanging ATPase ; metabolism

Objective To summarize the sonographic features and pathological features of ovarian tumor during pregnancy. Methods One hundred and five women with 114 pathologically proved ovarian tumors during pregnancy in Peking Union Medical College Hospital were retrospectively recruited. According to pathological diagnosis, the clinical treatment, the result of the pregnancy and sonographic examinations were reviewed and analyzed. The sonographic features of benign tumors were compared with low-grade malignant tumors. Results Among the 105 pregnant women with a total of 114 ovarian tumors, 65 tumors were found by ultrasound exam. The other 49 tumors were found during cesarean section. The sonographic features of pathologically proved ovarian tumors include regular shape and well-deifned margins, with 58 of benign tumors and 7 of borderline or low-grade malignant tumors. Compared with borderline or low-grade malignant tumors, benign tumors manifested as strong echoes or high echogenic mass without papillae in the tumors (50/58). As for borderline or low-grade malignant group, tumors manifested as papillae within the tumors (5/7). Pathological classiifcation of the 114 ovarian tumors included 84 germ cell tumors, 19 epithelial tumors, 9 sex cord-stromal tumors, and 2 germ cell tumors combined epithelial tumors. Surgical treatments were performed in 7 cases during the ifrst trimester, while 11 cases during the second trimester, and 87 cases during the third trimester. Pregnancy outcome of the 105 pregnant women included term delivery in 82 cases, premature delivery in 18 cases, artiifcial abortion during ifrst trimester in 4 cases, and induced abortion during second trimester in 1 case. Conclusions Most ovarian tumors treated in pregnancy are benign. The sonographic features of benign tumors include regular shape with well-deifned margins, strong echoes or high echogenic mass within the tumors. While the sonographic features of borderline or malignant tumors include papillae within the tumors. Ultrasound assessment of ovarian tumors can help to determine the risk of malignancy and guide the surgical management.

BACKGROUND:Dendritic cel s can regulate the immunological reaction in the intestinal tract, this functional deficit may induce inflammatory bowel disease. Tol-like receptor-4/nuclear factor-κB pathway is highly involved in this reaction. OBJECTIVE:To establish experimental colitis model in rats, to observe effects of resina draconis on dendritic cel s and Tol-like receptor-4/nuclear factor-κB expression in rats with experimental colitis, and to explore its action mechanism. METHODS:A total of 44 male Sprague-Dawley rats were randomly assigned to four groups (n=11):blank control group, model group, resina draconis group, 5-aminosalicylic acid treatment group. With the exception of blank control group, 2,4,6-trinitrobenzenesulfonic acid-induced ulcerative colitis models were established in the model group, resina draconis group and 5-aminosalicylic acid treatment group. After the models were successful y established, the rats in the resina draconis and 5-aminosalicylic acid treatment groups were intragastrical y treated with resina draconis [(0.75 g(kg·d)] and 5-aminosalicylic acid [100 mg(kg·d)] respectively for 10 days. RESULTS AND CONCLUSION:Disease activity index, macroscopic colonic damage score and histopathological score were significantly decreased in the resina draconis group compared with the model group (P<0.05). Symptoms and tissue damages were obviously lessened in the 5-aminosalicylic acid treatment and resina draconis groups compared with the model group. Expression rates of CD80 and CD86, as wel as expression levels of Tol-like receptor-4 and nuclear factor-κB were significantly higher in the model group compared with the blank control group, resina draconis group and 5-aminosalicylic acid treatment group (P<0.05, P<0.01). Tol-like receptor-4 and nuclear factor-κB expression was significantly lower in the resina draconis group than that in the 5-aminosalicylic acid treatment group. Experimental findings indicate that, resina draconis can partial y relieve experimental colitis symptoms in rats and effectively inhibit the activation of dendritic cel s in the mesenteric lymph node. Resina draconis can relieve enteric inflammatory reaction by suppressing the expression of Tol-like receptor-4 and nuclear factor-κB in rats.

Objective To evaluate the value of low-density lipoprotein-cholesterol (LDL-C) and non-high-density lipoprotein-cholesterol (non-HDL-C) in differential diagnosis of familial hypertriglyceridemia (FHTG) and familial combined hyperlipidemia (FCHL).Methods We recruited 9 FHTG pedigrees (94 subjects) and 24 FCHL pedigrees (94 subjects) and then divided them into affected groups and non-affected groups according to lipid abnormality.Another 10 normal control pedigrees (57 subjects) served as controls.We compared the routine lipid levels such as triglyceride (TAG),total cholesterol (TC),HDL-C and LDL-C and non-HDL-C between the groups.After stratification based on TAG level,we observed the relationship between LDL-C and non-HDL-C.Last we confirmed and analyzed the cut-off value of differential diagnosis between FHTG and FCHL with receiver operating characteristic (ROC) curve.Results The levels of TAG,TC,and non-HDL-C were significantly higher in the affected group of FHTG than in the non-affected group of FHTG and the normal group (P＜0.01 or P＜0.05).The levels of TAG,TC,HDL-C,LDL-C and non-tHDL-C wcrc significantly higher in the affected group of FCHL than in the non-affected group of FCHL and the normal group (P＜0.01 or P＜0.05).The levels of TAG were significantly higher (P＜0.01) while TC,HDL-C,LDL-C and non-HDL-C levels were significantly lower (P＜ 0.01 or P＜0.05) in the affected group of FHTG than in the affected group of FCHL.The association between LDL-C and non-HDL-C was positive both in FHTG and FCHL,but the relationship became weaker as TAG level increased.The cut-off value of LDL-C and non-HDL-C was 3.575 mmol/L and 4.525 mmol/L,respectively.Conclusion In addition to the routinely used lipid indexes,non-HDL-C may be a new index for differential diagnosis of FHTG and FCHL,and may be superior to LDL-C in this regard.

Objective To summarize the sonographic features and differential diagnosis points of mass-type cornual pregnancy. Methods The sonographic ifndings of 23 pathological proven mass-type cornual pregnancy cases enrolled in PUMCH from 2011 January to 2013 January were retrospectively analyzed. Results All pathological proven mass-type cornual pregnancy were located at one corner of the uterus presenting as a heterogenous outward mass. Well-deifned margins were found in 20 cases, and interstitial-line signs were found in 15 cases. The surrounding muscle thickness is 0.1-0.3 cm. Typical hyperechoic villi were found on sonography in cases with bloodβ-hCG>20 000 IU/L. On Doppler, the lesion showed abundant peripheral vascularity with low resistance in 22 cases, 9 lesions also showed abundant internal vascularity. Among 23 mass-type cornual pregnancy cases, 7 cases were misdiagnosed as gestational trophoblastic neoplasia (GTN) due to the similar sonographic characteristics including mixed-echo and abundant vascularity with low resistance. Sixteen cases were diagnosed by ultrasound preoperatively, with featured sonographic signs including mass located in the endometrial extension line;clear margin;peripheral vascularity;or detection of interstitial-line sign and typical villus. Conclusions Mass-type cornual pregnancy may be correctly diagnosed according to the location, boundary of the mass and the distribution of blood flow combining with clinical manifestation and bloodβ-hCG level. Transvaginal sonography could play an important role in diagnosis of cornual pregnancy.

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.