[Mechanism of nano-indium-tin oxide inducing pulmonary alveolar proteinosis in Sprague-Dawley rats].

Objective: To investigate the pathogenesis of pulmonary alveolar proteinosis in rats induced by nano-indium-tin oxide exposure, and to provide a basis for further determining the limit of occupational exposure to indium and developing related protection measures. Methods: In August 2018, a total of 40 specific pathogen-free Sprague-Dawley rats, with an age of 6-8 weeks and a body weight of (200±10) g, were randomly divided into control group, low-dose group (1.2 mg/kg) , middle-dose group (3 mg/kg) , and high-dose group (6 mg/kg) , with 10 rats in each group. After 1 week of routine feeding, the rats were given non-exposed intratracheal instillation twice every week, with an interval of 3 days, for 12 consecutive weeks. Body weight was measured every week during exposure to observe the change in body weight; The rats were anesthetized and sacrificed by chloral hydrate after the exposure ended, and lung tissue and serum were collected; Hematoxylin-eosin staining and periodic acid-Schiff (PAS) staining were performed for lung tissue to observe pathological results; Inductively coupled plasma mass spectrometry was used to measure the serum level of indium; ELISA was used to measure the levels of surfactant protein A (SP-A) , surfactant protein D (SP-D) , and the type II alveolar cell surface antigen Krebs von den Lungen-6 (KL-6) in lung tissue and the serum level of granulocyte-macrophage colony-stimulating factor (GM-CSF) . Results: The pathological results showed that the rats in the control group had basically complete alveolar structure, and after intratracheal instillation of nano indium-tin oxide, uniform, eosinophilic, and unstructured granular substances were observed in the alveolar space of the low-, middle-, and high-dose exposure groups, with macrophage proliferation and an increase in macrophages, especially in the high-dose group. Negative PAS staining was observed in the control group, while substances with positive PAS staining were observed in lung tissue in each exposure group. The three exposure groups had a significantly higher serum level of indium than the control group (P<0.05) . Compared with the control group, the three exposure groups had significant increases in SP-A, SP-D, and KL-6 in lung tissue and a significant reduction in GM-CSF in serum (P<0.05) . Conclusion: Pulmonary alveolar proteinosis in rats may be associated with the destruction of alveolar macrophages caused by nano-indium-tin oxide and the aggregation of pulmonary surfactants due to disorders in the metabolism and clearance of pulmonary surfactants by macrophages.

[Safety and efficacy of transcatheter closure of ruptured sinus of Valsalva aneurysm].

Objective: To investigate the safety and efficacy of transcatheter closure of ruptured sinus of Valsava aneurysm(RSVA). Methods: A total of 33 RSVA patients underwent transcatheter closure from January 2006 to March 2017 in our hospital were included in this retrospective study. The RSVA was diagnosed by echocardiography.Different type of occluders were applied for transcatheter closure based on the aortography results. All the patients were followed up after the procedure. Results: The patients were (37.6±12.1) years old,and the male patients accounted for 78.8%(26 cases).RSVA from right coronary sinus was found in 25 patients,and draining chamber was right atrium in 13 cases, right ventricle in 12 cases. RSVA from noncoronary sinus was diagnosed in 8 patients,and the draining chamber was right atrium. Aortography defined the narrowest diameter at the rupture site was (6.4±1.7)mm. The ratio of Qp/Qs was 2.2±0.5,and the mean pressure of pulmonary artery was 24.0(21.2,33.7)mmHg(1 mmHg=0.133 kPa). One patient developed serious occluder related aortic regurgitation and underwent surgery, transcatheter closure was successfully performed in 32 patients. The success rate of transcatheter closure was 97.0%. Two types of device were used in the study including small-waist double-disk ventricular septal defect(VSD) occluders in 20 cases and patent ductus arteriosus(PDA) occluders in 12 cases. During a median follow-up of 73.5(28.3,89.5) months, there were no infective endocarditis, residual shunt, thrombosis, device displacement,serious aortic regurgitation, serious arrhythmia or death.At the last follow-up, the left atrial diameter((37.4±6.5) mm vs. (41.5±5.3)mm,P<0.01),right atrial diameter((42.4±3.0) mm vs. (48.5±6.0)mm,P<0.01), right ventricular diameter((22.2±3.8) mm vs. (27.7±7.2)mm,P<0.01) and left ventricular end-diastolic diameter((51.3±4.9) mm vs.(55.0±4.3)mm,P<0.01)measured by echocardiography were all smaller than pre-procedural level. Conclusion: Transcatheter closure of RVSA is a safe and effective strategy and associated with a good long-term outcome.

[Effects of Blood-brain Barrier and Simulated metabolic system on Apoptosis of SH-SY5Y Induced by Acrylamide in Vitro].

Objective: To evaluate the effect of acrylamide on the apoptosis of nerve cells by integral cell modelling in vitro which simulates the barrier effect and metabolic micro-environment. Methods: A non-contact and co-cultured in vitro blood brain barrier (BBB) model was established by using human umbilical vein endothelial cells (HUVEC) and rat glioma cells (C6) . The trans-endotheilal electrical resistance (TEER) and horseradish peroxidase (HRP) tracer effects were measured to verify the tight connectivity and permeability of the established BBB model. An integrate discrete multiple organ cell co-culture (idMOC) model was established by inoculating the human renal cortical proximal tubular epithelial cells (HK-2) , human normal hepatocytes (L-02) and human neuroblastoma cells (SH-SY5Y) into the self-made multi-organ plate for co-culturing. Then the model was verified by observing the growth curve of various tissue cells under co-culturing or culturing individually. SH-SY5Y cells were exposed to different concentrations of acrylamide directly and indirectly (through BBB model and idMOC model) . The changes of cell apoptosis rate were analyzed by flow cytometry to explore the impact of model on Acrylamide (ACR) injury of typical neurotoxic agents. Results: HUVEC cells can form a wide range of close-connected complex and then inhibit the external electric field under the cross-endothelial movement, and the mean was lower than that of endothelial cell culture group at 4, 5 and 6 days (P<0.05) ; After 20 min, the penetration rate of HRP in the co-culture group was less than that in the individual culture group, and the difference was statistically significant (P<0.05) , indicating that the barrier function of the co-culture group was higher than that of the individual culture group. All cells can exchange substances through the exchange hole of the culture plate, the cells grow well and there was no obvious death. The growth curve in individual culture group and co-culture group were basically the same, the difference was not statistically significant (P>0.05) . Under the condition of different concentrations of ACR (140, 270 g/ml) , compared with the direct exposure group, the apoptosis rate of the BBB model and the idMOC model were significantly decreased, and the difference was statistically significant (P<0.05) . Conclusion: Based on HUVEC cells and C6 cells co-culture system, a blood-brain barrier model in vitro was established and based on co-culture of HK-2, L-02 and SH-SY5Y, the idMOC model was established. The toxicity and toxic action characteristics of ACR on SH-SY5Y cells were evaluated by validation tests.

[Therapeutic effects and related mechanisms of erythropoietin sustained-release gelatin hydrogel microspheres on a murine model of hindlimb ischemia].

OBJECTIVE: To investigate the therapeutic effects of erythropoietin sustained-release gelatin hydrogel microspheres (EPO-GHM) on a murine model of hindlimb ischemia and related mechanisms. METHODS: Fifty two ten weeks old male C57BL/6J mice were assigned to 5 groups: sham-operated group (the right femoral artery suture was passed through the right femoral artery but not tied, n=8); saline group (right femoral artery ligation and intramuscular injection of saline at a dose of 4 ml/kg into the right hind limb, n=12); EPO group(right femoral artery ligation and intramuscular injection of EPO at a dose of 5 000 IU/kg into the right hind limb, n=12), empty GHM group (right femoral artery ligation and intramuscular injection of empty GHM at a dose of 4 ml/kg into the right hind limb, n=8); EPO-GHM group(right femoral artery ligation and intramuscular injection of EPO-GHM at a dose of 5 000 IU/kg into the right hind limb, n=12). The blood flow ratio of ischemic limb (right)/nonischemic limb (left) was measured using a laser Doppler perfusion imager. After 8 weeks, immunohistochemical analysis were used to evaluate the vessel density (vessel density of CD31 positive), arteriole density(vessel density of α-smooth muscle actin(α-SMA) positive) and muscle area(HHF35 positive area). The proliferating index of vessels was evaluated by double immunofluorescent labeling to evaluate effect of EPO-GHM on angiogenesis of ischemia limb. Western blot was used to evaluate the protein expression of EPO receptor, protein kinase B(AKT), p-AKT, endothelial nitric oxide synthase(eNOS), p-eNOS and matrix metalloproteinase 2(MMP-2). RESULTS: (1) Eight weeks later, the blood flow ratio of ischemic limb/nonischemic limb was significantly higher in the EPO-GHM group compared with other groups(0.810±0.080, 0.563±0.051, 0.570±0.056 and 0.561±0.052 respectively, all P<0.05). (2) CD31 antibody positive and α-SAM antibody positive densities were higher in the EPO-GHM group compared with other groups(P<0.01 or 0.05). (3)HHF35 positive area in saline group, EPO group, empty GHM group and EPO-GHM group were smaller than that of sham-operated group(all P<0.05). HHF35 positive area in saline group, EPO group, empty GHM group and EPO-GHM group were similar(all P>0.05). (4)The proliferating index of vessels was higher in the EPO-GHM group compared with other groups(P<0.01 or 0.05). (5) Compared with other groups, the protein levels of EPO receptor, AKT, p-AKT, p-eNOS and MMP-2 were significantly higher in EPO-GHM group(P<0.01 or 0.05) and level of eNOS was similar among five groups(P>0.05). CONCLUSION: RESULTS from present study suggest EPO-GHM could improve blood perfusion of ischemia limb in mice through increasing capillary and arteriolar densities and these beneficial effects are possibly mediated by EPOR up-regulation and AKT/p-eNOS/MMP-2 signaling pathway activation.

Potential protective effect of nuclear factor-κB decoy oligodeoxynucleotides on endotoxin-induced liver injury.

PURPOSE: We sought to study the protective effects of nuclear factor-κB decoy oligodeoxynucleotides (ODNs) on endotoxin-induced liver injury in a rat model. METHODS: Sixty Sprague-Dawley rats were randomly divided into a control (n=20), a lipopolysaccharide (LPS) (n=20), and an NF-κB decoy ODN group (n=20). Liver and blood serum samples were collected at 24 hours after the operation. NF-κB binding activity was detected by an electrophoretic mobility shift assay, liver histopathology, by light microscopy; and cell apoptosis, by a terminal-deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay. The serum of liver enzyme (aspartate transaminase [AST]) levels were measured using an automated biochemical analyzer and tumor necrosis factor (TNF)-α and interleukin (IL)-6 by enzyme-linked immunosorbent assays. RESULTS: NF-κB was dramatically activated after endotoxin-induced liver injury. Many hepatocytes underwent degeneration and necrosis in the LPS group. The expressions of AST, TNF-α, and IL-6 were significantly increased compared with the control group (P=.0005), However, NF-κB decoy ODNs altered these undesirable changes. On the other hand, IL-6 expression was not significantly decreased by the NF-κB decoy versus the LPS group (P=.0745). CONCLUSIONS: NF-κB decoy strategy inhibited the binding activity of NF-κB, thus suppressing production of downstream cytokines which play crucial roles in protection from endotoxin-induced injury.

Disease association of the interleukin-18 promoter polymorphisms in Taiwan Chinese systemic lupus erythematosus patients.

Interleukin (IL)-18, an important mediator of innate and adaptive immunity, plays multiple roles in chronic inflammation, in autoimmune diseases, in a variety of cancers and in number of infectious diseases. IL-18 promoter polymorphisms have been also noted associated with various inflammatory diseases. We investigated the association of IL-18 promoter polymorphisms (-656T/G, -607A/C and -137C/G) with systemic lupus erythematosus (SLE) in Taiwan Chinese patients and controls. Six haplotypes (hts) were identified from the three promoter polymorphisms. The genotype distribution of the ht1 (GCC), ht2 (TAC), ht4 (GAC) and ht5 (TCC) were different in patients and controls (P<0.002). Moreover, the haplotype and genotype frequencies of ht1 were significantly increased in patients with discoid rash (P=0.045, odds ratio (OR): 2.01, 95% confidence interval (CI): 1.01-4.00; P=0.027, OR: 5.13, 95% CI: 1.41-18.68). In addition, the homozygous genotype ht1/ht1 was significant increased in patients with serositis (P=0.015, OR: 9.78, 95% CI: 1.55-61.73). These observations suggest that the three promoter polymorphisms contribute to the genetic background of SLE pathogenesis.

Secular change in the growth and development of Han children in China.

This paper reports the secular change in physical growth and development of Han children in 12 Chinese metropolitan cities including Beijing, Tianjing, Shanghai, Jinan, Harbin, Shenyang, Changchun, Nanjing, Hangzhou, Wuhan, Guangzhou, and Chengdu. Based on a recent national survey of 14,688 healthy schoolchildren, aged 7-18 years, and together using the historical records of healthy schoolchildren as secondary data source, this study found a secular change in the growth and development of Chinese children and youth. In every decade between the 1950s and 1985 the average height increased by 2.66 cm (range 1.78-3.77) for boys and 2.40 cm (range 1.72-3.76) for girls, and the average weight increased by 1.64 kg (range 1.28-2.63) for boys and 1.14 kg (range 0.63-2.01) for girls in the 12 cities. In the last decade from the 1970s to 1985, the height and weight increments were highest in Beijing, Jinan, Shenyang and Shanghai. In addition, the increments were higher during the peak growth years than during other periods of growth. In the 30 years studied, the ages of maximum growth velocity were advanced, and they were 1 or 2 years earlier in 1985 than in the 1950s in most cities. When compared with the data from the 1930s the results indicate that, during the last half-century, height increased by 1.12-2.66 cm per decade for boys and 1.42-2.67 cm/decade for girls, while weight increased by 0.56-1.27 for boys and 0.65-1.18 kg for girls in Beijing, Shanghai, Nanjing, Hanzhou and Guangzhou.