Objective:To investigate the effect of astragaloside A (AS-A) on the photoreceptor degeneration induced by sodium iodate (NaIO 3) and its related mechanism. Methods:Sixty healthy male C57BL/6J mice, aged 6-8 weeks, were randomly divided into normal control (NC) group, NaIO 3 group, and ASA group, with twenty mice in each group. 30 min before modeling, AS-A group mice were intraperitoneally injected with 100 μl AS-A at a dose of 100 mg/kg body weight. 30 min later, mice in NaIO 3 group and AS-A group were intraperitoneally injected with 100 μl NaIO 3 at a dose of 30 mg/kg body weight. Subsequently, AS-A group mice were administered AS-A twice daily at 12 h intervals until the end of the experiment. On day 1 post-modeling, zonula occludens-1 (ZO-1) immunohistochemistry was performed to observe the structure of retinal pigment epithelium (RPE) cells; real-time quantitative polymerase chain reaction (qPCR) was conducted to detect the mRNA expression of various retinal chemokine ligand-2 ( Ccl2), interleukin-1 beta ( Il-1β), mixed lineage kinase domain-like protein ( Mlkl), receptor-interacting protein kinase 3 ( Ripk3), and tumor necrosis factor ( Tnf). On day 3 post-modeling, immunohistochemistry was performed to observe the expression of ionized calcium binding adaptor molecule 1 (Iba1) and glial fibrillary acid protein (GFAP) in the retina; TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to detect photoreceptor cell death in each group. On day 4 post-modeling, fundus morphology of mice in each group was observed by fundus color photography and optical coherence tomography (OCT). Hematoxylin-eosin staining (HE) was used to observe the morphological structure of the retina in each group. Inter-group comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way ANOVA. Results:Fundus color photography and OCT examination showed that a large number of scattered yellow-white subretinal nodular structures in the fundus of NaIO 3 group mice, and a large number of strong reflection areas in the RPE layer. The number of strong reflection areas in the RPE layer was reduced in the AS-A group. Immunohistochemical analysis of ZO-1 showed that ZO-1 was largely lost on the RPE cell membrane in that NaIO 3 group; whereas in the AS-A group, ZO-1 was evenly distributed on the RPE cell membrane. HE staining results showed circular black deposits were visible in the RPE layer of the NaIO 3 group, and the inner and outer segments of photoreceptors were severely damaged, with a significant decrease in the number of outer nuclear layer (ONL) cell nuclei; whereas in the AS-A group, the RPE layer pigments were orderly, the inner and outer segments of photoreceptors were intact, and the number of ONL cell nuclei significantly increased. The results of TUNEL staining show that numerous TUNEL-positive cell nuclei were observed in the ONL of the retina in the NaIO 3 group, while the number of TUNEL-positive cell nuclei in the ONL of the retina was significantly reduced in the AS-A group, with statistically significant differences ( t=2.66, P<0.05). The analysis of qPCR data showed that compared with the AS-A group, the relative expression levels of Mlkl, Ripk3, Ccl2, Il-1β and Tnf mRNA in the retina were significantly increased in the NaIO 3 group, with statistically significant differences ( F=39.18, 10.66, 53.51, 41.40, 24.13; P<0.001). Immunohistochemical staining results showed that compared with NC group and AS-A group, the positive expression of GFAP in retina of NaIO 3 group was significantly increased, and the difference was statistically significant ( F=9.62, P<0.05). Conclusion:AS-A antagonizes NaIO 3-induced photoreceptor degeneration in part by inhibiting photoreceptor cell death and neuroinflammation. Meanwhile, AS-A treatment protects against NaIO 3-triggered perturbation of retinal homeostasis.

To investigate the effect and mechanism of metformin on intestinal epithelial barrier injury in ulcerative colitis. A cell model of colitis was established by co-culture of human colon cancer cell line Caco-2 and human monocyte cell line THP-1. The colitis model cells were treated with metformin at concentration of for Flow cytometry was used to detect Caco-2 cell apoptosis, and Western blotting was used to detect the protein expression of tight junction proteins and endoplasmic reticulum stress-related proteins. After metformin treatment, the apoptosis rate of Caco-2 cells was decreased from (14.22±2.34)% to 0.61)% (=3.119, <0.05), and the expression levels of tight junction protein-1 and claudin-1 increased (=5.172 and 3.546, both <0.05). In addition, the expression levels of endoplasmic reticulum-related proteins glucose regulated protein (GRP) 78, C/EBP homologous protein (CHOP) and caspase-12, as well as the phosphorylation level of PRKR-like endoplasmic reticulum kinase (PERK) and eukaryotic translation initiation factor 2α (eIF2α) decreased (all <0.05). Metformin may alleviate the intestinal epithelial barrier damage in colitis by reducing intestinal epithelial cell apoptosis and increasing the expression of tight junction proteins, which may be associated with the inhibition of endoplasmic reticulum stress-induced apoptotic pathway.
Apoptosis ; Caco-2 Cells ; Colitis, Ulcerative ; Endoplasmic Reticulum Stress ; Humans ; Metformin/pharmacology*

Objective: To establish the mouse aorta dissection (AD) model through drinking water containing β-aminopropionitrile (BAPN). Methods: Forty 3-week-old C57B1/6J male mice were divided into four groups according to randomized block design: control, 0.2, 0.4 and 0.8 g·kg^-1·d^-1 BAPN groups (dissolving respective dose of BAPN in the drinking water, n=10 each group). Arterial systolic blood pressure and heart rate were measured weekly in conscious, restrained mice using a noninvasive computerized tail-cuff system. Mice those died of rupture of aortic dissecting aneurysm during the study were autopsied and the aorta was examined. After 4 weeks, survived mice were sacrificed by an overdose of sodium pentobarbital and the whole aorta was harvested and analyzed. Results: The incidence of AD and the mortality of ruptured AD was 0 and 0 in control group, 30% (3/10) and 20% (2/10) in 0.2 g·kg^-1·d^-1 BAPN group, 50% (5/10) and 40% (4/10) in 0.4 g·kg^-1·d^-1 BAPN group, 90% (9/10) and 70% (7/10) in 0.8 g·kg^-1·d^-1 BAPN group (both P<0.05 vs. control group). The incidence of AD and the mortality of ruptured AD increased in proportion to BAPN concentration increase. In 0.8 g·kg^-1·d^-1 BAPN group, 7 mice died of dissecting aneurysm rupture during the experiment, among which 5 dissecting aneurysms were mainly located in the thoracic aorta and 2 dissecting aneurysms in abdominal aorta. The diameters of thoracic aorta and abdominal aorta were (1.38±0.19) and (1.23±0.13) mm in control group, (2.43±1.56) and (1.30±0.26) mm in 0.2 g·kg^-1·d^-1 BAPN group, (2.45±1.28) and (1.30±0.31) mm in 0.4 g·kg^-1·d^-1 BAPN group, (2.87±0.57) and (1.95±0.81) mm in 0.8 g·kg^-1·d^-1 BAPN group (both P<0.05 vs. control group). The diameters of thoracic aorta and abdominal aorta in mice also increased in proportion with BAPN concentration increase. Furthermore, blood-filled false lumen formation and elastic fibers fragmentation were evidenced in hematoxylin-eosin stained and Vitoria blue-Sirius red stained aortic cross-sections of mice in the 0.8 g·kg^-1·d^-1 BAPN group. Conclusion BAPN treatment induced aortic dissection model in C57Bl/6J mice can serve as a useful wild-type mouse model for the mechanism and pharmaceutical studies of AD.

Objective To investigate protective effect of tribulus terrestris L (TTL) on photoreceptor in the model of light-induced retinal degeneration.Methods BALB/c mice were exposed to bright light at the intensity of 10 000 lux for 30 minutes to establish the retinal light damage models.The BALB/c mice were divided into normal control group,model group and treatment group,6 cases in each group.TTL decoction was intraperitoneally administered to mice 30 minutes prior to illumination in the treatment group.Saline vehicle was administered in the normal control group and model group.Photoreceptor protection of TTL was assessed by optical coherence tomography (OCT) at 3 hours and 7 days after illumination.Gross histology and immunohistochemistry approaches were also taken to examine the retinal protection conferred by TTL at 7 days after bright light exposure.Results Compared to normal retinal morphology in the normal control group,prominent photoreceptor loss and diminished rod and cone photoreceptors evidenced by attenuated retinal expression of rhodopsin and M-opsin were observed in the model group.In contrast,TTL treatment resulted in significant protection against bright light-induced photoreceptor degeneration and remarkable preservation of rod and cone photoreceptor cells.The outer retinal nuclear layer in the model group was thinner than that in the normal control group (P ＜ 0.05),but the treatment group was thicker than the model group (P ＜ 0.05).Conclusion Bright light induces obviously degeneration in photoreceptors in BALB/c mice.Moreover,TTL is shown for the first to significantly protect the photoreceptors from bright light-induced degeneration.

Objective To establish a mouse model of aorta dissection (AD) by β-aminopropionitrile (BAPN) in drinking water + subcutaneously pumped angiotensin II (Ang II) infusion.Methods Forty 3-week-old C57B1/6J male mice were randomly divided into two groups.All animals received 0.1 g/kg/d BAPN in drinking water for 4 weeks.Then the BAPN drinking + saline infusion group and BAPN drinking + Ang II infusion group received continuous saline or Ang II (1,000 ng/kg/min) infusion, respectively, via subcutaneous osmotic minipump for 72 hour.The mice were restricted in a noninvasive computerized tail-cuff system and their arterial systolic blood pressure and heart rate were monitored.Autopsy was performed if a mouse died during the experiment.At the end of the experiment, mice were sacrificed by injection with an overdose of sodium pentobarbital and the aortas were harvested.The formation of aortic false lumen was observed by pathology using hematoxylin-eosin staining.Results The overall incidence of AD in the BAPN drinking administration +Ang II infusion group was 95%, whereas the incidence of AD in the BAPN drinking administration +saline infusion group was only 5%.The mortality from dissecting aneurysm rupture was 24% in the BAPN drinking administration +Ang II infusion group during the experiment.Pathological examination of the aortic cross-sections clearly showed the formation of blood-filled false lumens induced by Ang II.Conclusions A mouse model with high incidence of aortic dissection is successfully established.

Objective To investigate the relationship between plasma albumin and hemoglobin(Hb) levels and prognosis in patients with severe craniocerebral trauma. Methods The clinical data of 124 patients with severe craniocerebral trauma form January 2010 to January 2014 were analyzed retrospectively. The relationship between plasma albumin and Hb levels and prognosis were analyzed. Hb level was obtained in 3 days of admission, and then the patients were divided into Hb <90 g/L group, Hb 90-99 g/L group, Hb 100-110 g/L group, Hb >110 g/L group, and also were albumin <25 g/L group, albumin 25-28 g/L, albumin 29-31 g/L group, albumin>31 g/L group according to the mean albumin level. Multifactor Logistic regression analysis was used to analyze the relationship between Hb, albumin levels and prognosis. Results Among 124 patients, 37 patients (29.8%) were given red blood cell (RBC) transfusion, and 28 patients (22.6%) received albumin treatment. The hospital mortality was 20.2% (25/124). The age, scores of Glasgow Coma Scale (GCS) and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ ), and hospital mortality in different Hb level groups had no significant differences (P>0.05). The age, scores of GCS score and APACHEⅡ score in different albumin level groups had no significant differences (P>0.05), but the hospital mortality in different albumin level groups had significant difference: 31.2%(10/32), 24.2%(8/33), 9.7%(3/31), 14.3%(4/28), P<0.05. Multifactor Logistic regression analysis showed that albumin level was the relevant factor for the severe craniocerebral trauma (P<0.01), the hospital mortality was decreased with the rising of the albumin level, but the risk of death was higher in albumin>31 g/L group than that in albumin 29-31 g/L group. Conclusions 29-31 g/L albumin level is the best for severe craniocerebral trauma patients. There is no significant impact of Hb on the prognosis for severe craniocerebral trauma patients.

Objective To investigates the feasibility of diffusion tensor imaging (DTI) and diffusion tensor tracking (DTT) for evaluation of the effect of childbearing history on female pelvic floor muscles. Methods Forty-six healthy females were divided into two groups: nulliparous and primiparous. MR conventional sequences and DTI were acquired. The optimized FA threshold value was obtained by regulating the FA to fiber tracking. The two groups were compared in terms of ADC, FA, VRA and T2-WT. Results (1)The DTT of FA 0.18 got the highest score in fiber tracking . ( 2 ) The ADC of nulliparous subjects and the subjects who had given birth were (1.24 ± 0.11) ×10-3 mm2/s, (1.33 ± 0.11) ×10-3 mm2/s (P = 0.017). There were no statistical differences in FA, VRA and T2-WT between the two groups (P > 0.05). Conclusions The optimized FA threshold of fiber tracking in pelvic floor muscles is 0.18. DTI and DTT may be used to evaluate the effect of childbearing history on female pelvic floor muscles.

Objective To compare four pretreatment methods of human plasma coagulation factor Ⅷ( FⅧ) before column purification.Methods Cryoprecipitate was dissolved in Tris , before the supernatant was treated with glycine precipitation, PEG precipitation, acid precipitation and aluminum hydroxide gel adsorption , respectively.Activated partial thromboplastin (APTT) was used to measure the activity of the supernatant clotting FⅧ after treatment.The total activity recovery and specific activity of the final samples were used to weigh the efficacy of those methods .The purity of the intend-ed protein was estimated by non-reducing SDS-PAGE electrophoresis .Results Total activity recovery of glycine precipitati-on was the highest (94.00%±7.60%), followed by that of acidic precipitation (89.47%±2.60%) and PEG precipita-tion (80.92%±9.67%) methods.The lowest was aluminum hydroxide gel adsorption (78.65%±7.52%).Glycine precipitation and PEG precipitation could more effectively remove contaminating protein than acid precipitation and aluminum hydroxide gel adsorption .Treated by four different methods , the specific activity of FⅧ of glycine precipitation sample was the highest (0.6856 ±0.1258 IU/mg), followed by PEG precipitation (0.5773 ±0.0787 IU/mg) and acidic precipitation (0.3885 ±0.0301 IU/mg).The specific activity of aluminum hydroxide gel adsorption was the lowest (0.2879 ±0.0472 IU/mg).Conclusion PEG precipitation is more effective for the actual production process than the other three methods .

Objective To evaluate the differences of the lower extremity atherosclerosis between patients with and without type 2 diabetes using dual-source CT angiography.Methods Dual-source CT angiography of lower extremity was performed in 87 patients with (n=30)or without (n= 57 )diabetes.Extent of luminal stenosis,and the type,distribution and range of the plaques were compared.Results 342 plaques in 540 segments (63.3%)in diabetic patients,and 500 plaques in 1 026 segments (48.7%)in non-diabetic ones were detected respectively.Compared with non-diabetic patients,the diabetic ones had a higher overall incidence of plaques (P <0.05).Calcified plaques were the most common in both kinds of patients,and the incidence of mixed plaques was high-er in diabetic patients than that in non-diabetic ones (35.6 % vs.28.4%,P <0.05).Light to moderate stenosis occurred in most diabetic patients,and fewer occlusion was found compared with non-diabetic ones (9.1% vs.1 7.0%,P <0.05).The most common sites of the plaques in diabetic patients were located at distal small arteries below the knee.However,those were located at proximal arteries above the knee for non-diabetic ones.The involvement of atherosclerosis in diabetic patients was more diffused,and the de-gree of Ⅳ (75%-100%)was higher than that in non-diabetic ones (P <0.05).Conclusion Atherosclerosis in lower extremity on dual-source CT angiography is very common in diabetic patients with multi-segmental,diffused,non-obstructive involvement of dis-tal small arteries below the knee.

Objective As a collaborator of Beijing Institute of Medical Device Testing for value assignment of state standard ma-terial candidate Cystatin-C ,we have used the internationally accepted reference material to assign value for state standard material candidate Cystatin-C ,and help Beijing Institute of Medical Device Testing get Cystatin-C national standard material certificate . Methods According to the target value and operational procedure of international reference material ERM-DA471 ,We have tested 6 dilutions of standard material candidate Cystatin-C on calibrated Hitachi 7180 immunoassay system .Results The results demon-strate good repeatability and commutability ,and have been accepted in calculating the final value for the candidate standard materi-al ,our data has assisted Beijing Institute of Medical Device Testing in passing the criteria and obtaining Cystatin-C national standard material certificate .Conclusion Compared to the data from all participating collaborators ,our results hit right on the target value , and no significant matrix effects have been observed .