OBJECTIVETo determine whether quercetin inhibits oxidized low-density lipoprotein (Ox-LDL)-induced osteogenic differentiation and calcification of vascular smooth muscle cells (VSMCs) and understand the underlying mechanism.

METHODSThe calcification of human VSMCs following Ox-LDL treatment was assessed using alizarin red staining and by detecting ALP activity. The mRNA expressions of the bone-related genes including Msx2, BMP2 and Osterix, and the contractile proteins including SMA and SM22a were analyzed using qPCR. The effects of quercetin were investigated on OxLDL-induced VSMC calcification and changes in ALP activity, expressions of Msx2, BMP2, Osterix, SMA and SM22a, ROS levels and SOD activity. The effect of Toll like receptor 4 (TLR4) silencing mediated by siRNA transfection on cell calcification, ALP activity, gene expressions and ROS levels were investigated.

RESULTSOx-LDL treatment promoted VSMC calcification and up-regulated TLR4 expression. Quercetin treatment significantly attenuated Ox-LDL-induced VSMC calcification, reduced ALP activity, down-regulated the expression levels of Msx2, BMP2 and Osterix, and up-regulated the expressions of vascular smooth muscle contractile proteins SMA and SM22a. In addition, Quercetin treatment markedly increased SOD activity, reduced ROS levels and TLR4 expression in VSMCs. Silencing TLR4 expression using TLR4 siRNA also significantly decreased calcification of the VSMCs.

CONCLUSIONSQuercetin inhibits Ox-LDL-induced VSMC calcification in VSMCs possibly by targeting the ROS/TLR4 signaling pathway.

Objective To explore the characteristics of respiratory parameters in patients with different body mass index during general anesthesia with tracheal intubation. Methods 102 patients scheduled for otitis me-dia surgery were divided into low weight group(B1,n=32),normal weight group(B2,n=36)and overweight or obese group(B3,n = 34 ). After general anesthesia with tracheal intubation,the tidal volume of anesthetic ma-chine wasadjusted to maintain the end tidal carbon dioxide partial pressure between 35 - 45 mmHg. At 10 min (T1),30min(T2)and 60 min(T3)after adjustment,arterial PH,arterial partial pressure of oxygen(PaO2),arte-rial carbon dioxide pressure(PaCO2),inspiratory tidal volume(VTi),expiratory tidal volume(VTe),end tidal carbon dioxide partial pressure(PETCO2),peak airway pressure(Ppeak),plateau airway pressure(Pplat)and dy-namic lung compliance(Cdyn)were recorded. Results PH and PaO2 were not significantly different at T1-3 among the three groups(P>0.05). As compared with group B1 and B2,PaCO2 was lower in group B3. In comparison with group B2,VTi,VTe and Cdyn were higher in group B1 and lower in group B3(P < 0.05). Ppeak and Pplat were lower in group B1 but higher in group B3(P<0.05). PETCO2 was higher in group B1(P>0.05)while lower in group B3 (P < 0.05). Conclusions With the increase in BMI during general anesthesia with tracheal intubation ,the VTi,VTe,Cdyn,PETCO2 and PaCO2 decrease significantly,but Ppeak and Pplat elevate markedly. BMI is a refer-ence index for setting respiratory parameters.

Objective To summarize the clinical experience of patient diagnosed with Niemaoh-Pick disease being pregnant after liver transplantation. Methods Clinical data of one case of type B Niemaoh-Pick disease being pregnant after liver transplantation were retrospectively analyzed. Results The patient successfully underwent liver transplantation combined with splenic artery ligation on July 8, 2011. She was well recovered postoperatively. After surgery, she received conventional anti-rejection treatment, and gradually switched to use of tacrolimus at a dosage of 2.5 mg/d. The serum drug concentration was maintained at 2 ng/mL. In September 2015, she was successfully pregnant. On June 2, 2016, she delivered a male infant through cesarean section. She could breastfeed the infant in a low quantity early after delivery. Both the mother and infant were followed up until submission date. The mother was physically stable and the infant grew normally. Conclusions Patients diagnosed with Niemaoh-Pick disease can obtain favorable clinical outcomes of pregnancy and delivery after liver transplantation.

Objective To investigate the expression of Dickkopf-3 (DKK3) in human malignant melanoma cell lines and tissues,and to evaluate effects of DKK3 on the proliferation and apoptosis of malignant melanoma cell line A375.Methods Reverse transcription PCR (RT-PCR) and real-time fluorescence-based quantitative PCR (qRT-PCR) were performed to measure the mRNA expression of DKK3 in human malignant melanoma cell lines HM,A375,WM451,WM35,SK-MEL-1,Hs-695T and MDA-MB-435s,as well as in 38 primary melanoma tissues,4 metastatic melanoma tissues and 20 pigmented nevus tissues.Cultured malignant melanoma A375 cells were divided into 2 groups to be transfected with pcDNA3.1 (+)-Flag-DKK3 (experiment group) and pcDNA3.1 (+)-Flag-Vector (control group) respectively.The overexpression of DKK3 was verified by RT-PCR.Cell counting kit-8 (CCK8) assay and plate colony formation assay were performed to evaluate the proliferative activity of A375 cells,flow cytometry was conducted to detect apoptosis of A375 cells,and Western blot analysis was performed to determine the expression of cell cycle-and cell apoptosis-related proteins.Results The mRNA expression of DKK3 was downregulated in WM35 cells,absent in HM cells,A375 cells,WM451 cells,SK-MEL-1 cells and Hs-695T cells,but upregulated in MDA-MB-435s cells.Compared with pigmented nevus tissues,the mRNA expression of DKK3 was significantly decreased in malignant melanoma tissues (P ＜ 0.001).Compared with the control group (100％),cell colony formation was markedly suppressed in the experiment group (23.22％ ± 3.55％),and the proliferative activity of A375 cells was also significantly inhibited in the experiment group 24,48,72 hours after the transfection (all P ＜ 0.05).Flow cytometry showed that compared with the control group,A375 cells were significantly arrested in G1 phase (48.68％ ± 3.92％ vs.25.38％ + 2.92％,P ＜ 0.001),and the apoptosis rate of A375 cells was significantly increased in the experiment group (P ＜ 0.001).Compared with the control group,the experiment group showed significantly higher expression of p21,Bax,cleaved-parp and cleaved-casp3,but significantly lower expression of cyclin D1 and Bcl2 (all P ＜ 0.001).Conclusion DKK3 expression is downregulated in human malignant melanoma tissues,so it may serve as a potential tumor suppressor gene involved in the development of cutaneous malignant melanoma.

Objective To analyze the correlation between clinical features and renal dysfunction in patients of acute lacunar infarction with progressive cerebral microbleeds (CMBs). Methods Two hundred and sixty-five patients with first-episode acute lacunar infarction were selected. The serum creatinine was measured within 24 h of admission and the estimated glomerular filtration rate (eGFR) was calculated. The brain MRI (including gradient-echo images) was examined within 2 d of admission and after 1 years of follow-up, respectively. The progressive CMBs was assessed with microbleeds anatomical rating scale (MARS), and the patients were divided into progressive CMBs group (progressive group, 42 cases) and non progressive CMBs group (non progressive group, 223 cases). The clinical features of 2 groups were compared and the correlation between progressive CMBs and renal dysfunction was analyzed. Results The age, 24 h pulse pressure, incidences of renal dysfunction and CMBs in progressive group were significantly higher than those in non progressive group: (69.8 ± 5.8) years vs. (61.5 ± 4.9) years, (63.3 ± 3.1) mmHg (1 mmHg=0.133 kPa) vs. (51.8 ± 4.2) mmHg, 69.0％(29/42) vs. 39.9％(89/223) and 57.1％(24/42) vs. 25.1％(56/223), and the platelet was significantly lower than that in non-progression group:(168 ± 35) ×109/L vs. (189 ± 40) ×109/L, and there were statistical differences (P<0.05 or<0.01). The Logistic regression analysis result showed that renal dysfunction and CMBs were Independent risk factors of progressive CMBs (OR = 1.571 and 1.054, 95％ CI 1.042 - 2.493 and 1.010 - 1.142, P<0.05). Conclusions The rate of renal dysfunction is higher in patients of acute lacunar infarction with progressive CMBs, and progressive CMBs are associated with renal dysfunction.

Objective To investigate the relationship between different topographic locations and neurological deteriorations (ND) in patients with acute new isolated pontine infarction.Methods One hundred sixty-eight patients with acute new isolated pontine infarction during arch 2012 to March 2016 were identified by diffusion weighted imaging (DWI) for retrospective review.Patients were divided into two groups according to their clinical symptoms:patients with ND and patients without ND.According to neuroimaging of DWI,the topographic location of pontine infarction was divided into three types:The upper,middle,and lower ones,and the correlations of ND with risk factors,laboratory examination results,clinical manifestations and different topographic locations were explored by statistical tests.Results Of 168 patients,26.8％ (45/168) were diagnosed with ND,and 73.2％ (123/168) were diagnosed without ND.Univariate analysis showed that there were differences in female ratio [62.2％ (28/45) vs 41.5％ (51/ 123)],smoking ratio [13.3％ (6/45) vs 26.0％ (32/123)],mean length of hospital stay [(22.83 ± 7.12)d vs (19.31 ± 7.65)d],ratio of worse short-term clinical outcomes [77.8％ (35/45) vs 33.3％ (41/123)],and ratio of lower pontine infarction [55.6％ (25/45) vs 26.0％ (32/123)] between two groups (P ＜ 0.05).Logistic regression analysis showed that lower pontine infarction was the independent risk factor of ND (OR =1.953,95％ CI:1.092-3.535,P =0.029).Conclusions Topographic location of lower pons lesions may be reliable predictor of ND in acute new isolated pontine infarction.

Objective To investigate the diffevential diagnostic value of preinvasive and invasive lung adenocarcinoma (including minimally invasive adenocarcinoma and invasive adenocarcinoma) presented as pure ground-glass nodules(pGGN) by CT.Methods One hundred and fifty-six cases of pGGN verified by operative pathology were retrospectively analyzed,including 58 ca ses of preinvasive adenocarcinoma and 98 cases of invasive adenocarcinoma(TNM staging were T1N0M0).The CT features and sex were statistically processed.The difference between the CT features and sex were performed by thex2 test.The ROC curve of lesion focus size was drawn.Results Statistically significant differences were found in the lesion shape,vacuole sign,air bronchogram,blood vessel through,tumor-lung interface and vascular cluster sign between the two groups(all P＜0.05).The ROC curve showed that the accuracy rate of invasive adenocarcinoma was 75.0％ when the size of the pGGN lesions was larger than 15.35 mm.Conclusion The lesion size,shape,vacuole sign,air bronchogram,blood vessels through and vascular cluster sign have some predictive value.

To explore the clinical characteristics, imaging manifestation, diagnosis and treatment for histoplasmosis and to improve therapeutic level, we retrospectively analyzed the clinical data of 8 patients with biopsy-confirmed histoplasmosis from 2004 to 2014 in the Second Xiangya Hospital of Central South University and reviewed relevant literatures. The main clinical symptoms of histoplasmosis included fever, cough, expectoration, chest pain, blood-stained sputum, lymphadenectasis, etc. The major lung imaging features were mass, node or pneumonia-like performance. No case was diagnosed as histoplasimosis firstly. Four patients whose imaging manifestations were focal pulmonary lesion received lobectomy of lung lesions or wedge resection. Clinical and imaging manifestations in 3 patients, who treated with amphotericin B or its liposomal, itraconazole or fluconazole, were improved. The clinical symptoms and imaging findings of histoplasmosis are nonspecific. It is easy for the physicians to misdiagnose histoplasmosis as bacterial infection, lung cancer, tuberculosis lymphoma, etc. Therefore, it is significant and necessary to carry out multiple biopsies combined with multiple etiological examinations for patients with difficult diagnosis.
Amphotericin B ; Biopsy ; Diagnostic Errors ; Histoplasmosis ; Humans ; Lung ; Lung Neoplasms ; Pneumonia ; Retrospective Studies ; Sputum

BACKGROUND:Previous studies on immunosuppression and anti-rejection after organ transplantation mainly focused on effects of T lymphocytes-mediated immune response and immunosuppressive agents on T lymphocytes. Effects of dendritic cel s were unclear. The manifestation and mechanism of immunosuppressive agent effects on dendritic cel s are not identical. OBJECTIVE:To compare the effects of different immunosuppressive agents on expression and function of costimulatory molecules of dendritic cel s, and to explore the mechanism of action of immunosuppressive agents. METHODS:20μg/L rapamycin, 0.04 mg/L mycophenolate, 10μg/L tacrolimus and 1 mg/L cyclosporine A were separately added during bone marrow cel s of C57BL/6 mice were differentiated into dendritic cel s. RESULTS AND CONCLUSION:Flow cytometry results revealed that CD40 expression in each group:rapamycin0.05). One-way mixed lymphocyte reaction results displayed dendritic cel costimulatory T cel proliferation in each group:rapamycin

Objective To observe the effects of Taizhian treatment on macrophages expression in the plaque of aotic intima and blood lipid levels in hypercholesterolemic rabbits. Methods Sixteen male New Zealand rabbits were given 1%high-cholesterol diet. After 8 weeks,these macrophages expression in aotic intima was detected by immunohistochemistry. Results Compared with normal rabbits,Taizhian treatment decreased the expression of macrophages in aortic intima by 25%and reduced serum levels of low density lipoprotein-cholesterol by 21%in hypercholesterolemic rabbits(both P<0.05). Conclusions Taizhian may significantly decrease blood lipid level and reduce atherosclerosis in hypercholesterolemic rabbits.