Objective: To construct mcpr1prokaryoti c expression vector and to express MCPR1 protein.Methods:PCR was used to obtain coding region of mcpr1. Construction of a high-level fusion protein expression vector pGEX-4T-mcpr1 was conducted by inserting the fra gment of coding region of mcpr1into a fusion protein expression vector pGEX -4T-1. Then the recombinant plasmid was transferred into E. colito prepar e the MCPR1/GST fusion protein. DNA sequencing and endonucleases digesting were used to check the coding region. Results:pGEX-4T- mcpr1 wa s constructed successfully and the coding region was inserted into the vector co rrectly. A new protein band of 36 000 was observed by SDS-PAGE analysis after i nduction by IPTG. The 36 000 protein amounted to 39 percent of the total prote in and existed mostly in precipitation of broken bacteria. Conclusion: MCPR1 protein can be expressed in E. coliexpression system and purif ied initially.

Background Cystoid macular edema(CME) is an important cause of visual impairment of central retinal vein occlusion(CRVO).Spectral-Domain optical coherence tomography(SD-OCT) has increased speed and higher resolution,offering a better chance of understanding the morphological changes and pathogenesis of CME. Objective This study was to survey the morphologic features of macular edema associated with CRVO by SD-OCT. Methods Clinical data of the patients with CRVO diagnosed in Department of Ophthalmology,Peking Union Medical College Hospital from March 2008 to August 2009 were retrospectively analyzed.SD-OCT features of macular edema induced by CRVO were analyzed and recorded.Results The average macular foveal thickness was(527.5±218.2) μm in macular edemas eyes.Main morphological changes included 55 cases(84.6%) of CME,15 cases of(23.1%) serous macular detachment(SMD),and 10 cases(15.4%) of simple macular edema,and these findings occurred at the same time in some eyes.Cystoid spaces in the parafoveal region were seen in the inner nuclear layer,outer plexiform layer and outer nuclear layer,and discontinuous or weak inner segment/outer segment(IS/OS) line was often seen in CME.The incidence of CME associated with incomplete posterior vitreous detachment(PVD) was 14.5%,and that of neural epithelial edema associated with incomplete PVD was 10.0%,showing an insignificant difference between them(χ2=0.000,P=1.000).The average area of SMD was 1838.4μm ×1428.1μm×190.1μm,and the incidence of partial PVD was higher(χ2=4.266,P=0.039).Conclusion SD-OCT can reveal the micro-morphological change of macular zone in macular edema eye.SD-OCT enabled visualization of its spatial extent in each retinal layer and the condition of IS/OS layer.Serous macular edema is related with partial PVD.

BACKGROUND: In recent years, the immunoregulatory effects of mesenchymal stem cells (MSCs) can be used to induce immune tolerance. OBJECTIVE: To evaluate the safety and feasibility of human umbilical cord-derived MSCs (hUC-MSCs) in liver transplantation patients. METHODS: hUC-MSCs were cultured and identified. After approved by the Medical Ethics Committee, a total of 50 patients were randomly divided into experimental group and control group according to the proportion of 1:1. In the experimental group, hUC-MSCs were perfused by the portal vein during the operation and infused into the jugular vein on the 3rd day after the operation. The injection dose was 1×106/kg (prepared as 50 mL of cell suspension). Both groups received standard immunosuppressive regimens. Blood biochemistry and immune function indicators were detected preoperatively and at postoperative days 3, 7, months 1, 2, 3, 6, 12. Acute and chronic rejection rates, incidence of infection, and incidence of transplantation-related complications were recorded. RESULTS AND CONCLUSION: (1) At 3 and 7 days after the operation, the percentage of peripheral blood CD4+CD25+ cells (regulatory T cells) in the experimental group was significantly higher than that in the control group (P < 0.05). The percentage of CD4+ cells (helper/inducer T cells) and ratio of CD4+/CD8+ T lymphocytes were significantly lower in the experimental group than the control group (P < 0.05). (2) There was no significant difference in postoperative alanine aminotransferase and total bilirubin levels between the two groups (P > 0.05). (3) The incidence of abnormal liver function in the experimental group was significantly lower than that in the control group (P < 0.05). (4) The incidence of transplantation-related complications and the rate of infection showed no significant difference between the two groups (P > 0.05). Overall, the intravenous infusion of hUC-MSCs is safe and feasible in liver transplantation patients, which in early stage can promote the the proliferation and activation of CD4+CD25+ cells (regulatory T cells), reduce the percentage of CD4+ cells (helper/inducer T cells) and lower the ratio of CD4+/CD8+ T cells, thereby improving the immune status in liver transplantation patients.

The association of serum nesfatin-1 levels with insulin resistance and pancreatic β-cell function in gestational diabetes mellitus was investigated.Oral glucose tolerance test(OGTT) was performed in ninety pregnant women from 24,to 28 gestational weeks.They were divided into three groups according to OGTT:45 nomal controls,27 gestational diabetes mellitus with fasting plasma glucose (FPG) of 5.1 mmol/L to 7.0 mmol/L (GDM1),18 gestational diabetes mellitus with FPG more than 7.0 mmol/L (GDM2).Serum nesfatin-1 levels were significantly higher in patients with GDM1 and GDM2 than in controls (P＜0.01),and in GDM2 group it was also higher than GDM1 group(P＜0.05).Fasting serum nesfatin-1 was positively correlated with FPG,30 min plasma glucose,1 h plasma glucose,2 h plasma glucose,homeostasis model assessment for insulin resistance,and PGAUC,but negatively correlated with homeostasis model assessment for β-cell function.Furthermore,multiple stepwise regression analysis showed that FPG was the independent influencing factor of serum nesfatin-1 level.Nesfatin-1 was positively correlated with insulin resistance,while negatively correlated with pancreatic β-cell function.Nesfatin-1 may play a role in the pathogenesis of gestational diabetes mellitus.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Histone-Lysine N-Methyltransferase ; genetics ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Transcription Factors ; genetics ; Young Adult

The effect of astaxanthin on N(Ω)-nitro-L-arginine methyl ester (L-NAME) induced preeclampsia disease rats was investigated. Thirty pregnant Sprague-Dawley rats were randomly divided into three groups (n = 10): blank group, L-NAME group and astaxanthin group. From day 5 to 20, astaxanthin group rats were treated with astaxanthin (25 mg x kg(-1) x d(-1) x bw(-1)) from pregnancy (day 5). To establish the preeclamptic rat model, L-NAME group and astaxanthin group rats were injected with L-NAME (125 mg x kg(-1) x d(-1) x bw(-1)) from days 10-20 of pregnancy. The blood pressure and urine protein were recorded. Serum of each group was collected and malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide synthase (NOS) activities were analyzed. Pathological changes were observed with HE stain. The expression of NF-κB (nuclear factor kappa B), ROCK II (Rho-associated protein kinase II), HO-1 (heme oxygenase-1) and Caspase 3 were analyzed with immunohistochemistry. L-NAME induced typical preeclampsia symptoms, such as the increased blood pressure, urinary protein, the content of MDA, etc. Astaxanthin significantly reduced the blood pressure (P < 0.01), the content of MDA (P < 0.05), and increased the activity of SOD (P < 0.05) of preeclampsia rats. The urinary protein, NO, and NOS were also decreased. HE stain revealed that after treated with astaxanthin, the thickness of basilal membrane was improved and the content of trophoblast cells and spiral arteries was reduced. Immunohistochemistry results revealed that the expressions of NF-κB, ROCK II and Caspase 3 in placenta tissue were effectively decreased, and HO-1 was increased. Results indicated that astaxanthin can improve the preeclampsia symptoms by effectively reducing the oxidative stress and inflammatory damages of preeclampsia. It revealed that astaxanthin may be benefit for prevention and treatment of preeclampsia disease.
Animals ; Blood Pressure ; Caspase 3 ; metabolism ; Disease Models, Animal ; Female ; Heme Oxygenase (Decyclizing) ; metabolism ; Malondialdehyde ; metabolism ; NF-kappa B ; metabolism ; NG-Nitroarginine Methyl Ester ; Nitric Oxide Synthase ; metabolism ; Oxidative Stress ; Placenta ; enzymology ; Pre-Eclampsia ; drug therapy ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Xanthophylls ; therapeutic use ; rho-Associated Kinases ; metabolism

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.
Color ; Fabaceae ; chemistry ; enzymology ; growth & development ; metabolism ; Germination ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; metabolism ; Starch ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

This study assessed the clinical application of transvaginal three-dimensional ultrasound (3D TVUS) in the diagnosis of congenital uterine malformation. A retrospective study was performed on 62 patients with congenital uterine malformation confirmed hysteroscopically and/or laparoscopically. The patients were subjected to transvaginal two-dimensional ultrasound (2D TVUS) and 3D TVUS. The accuracy rate was compared between the two methods. The accuracy rate of 3D TVUS was (98.38%, 61/62), higher than that of 2D TVUS (80.65%, 50/62). 3D TVUS coronal plane imaging could demonstrate the internal shape of the endometrial cavity and the external contour of the uterine fundus. It allowed accurate measurement on the coronary plane, and could three-dimensionally show the image of cervical tube, thereby providing information for the diagnosis of some complex uterine malformation. 3D TVUS imaging can obtain comprehensive information of the uterus malformation, and it is superior to 2D TVUS for the diagnosis of congenital uterine malformations, especially complex uterine anomaly.

Objective To establish an animal model for implantable ventricular assist device. Methods Seven male calves, body weight 120 to 180 kg, were anesthetized and underwent surgical implantation through a left thoracotomy at the 4th intercostal space. The domestic-made implantable ventricular assist device (DIVAD) was implanted after cardiopulmonary bypass and induction of ventricular fibrillation. The major indexes of mechanical properties of the DIVAD are similar to those of imported devices. The DIVAD was 29.5 mm × 72 mm, with a total weight of 158 g. The maximal pump speed was 9000 rpm and the maximal pump flow was 8 L/min in in vitro experiment. A flow meter was incorporated into the pump. The DIVAD was constructed with an external controller, and the pump can be monitored and adjusted through it. The inflow cannula was positioned into the left ventricle and the outflow cannula was sutured to the descending aorta. The pump was run at 3,500 to 8,000 rpm. The cardiopulmonary bypass was weaned. The pump output and animal condition was continuously monitored. Heparin was administered to maintain the activated clotting time at 1.5 to 2 times its baseline value. Results All the seven calves weaned from cardiopulmonary bypass. The mean survival time is 20.28 hours with a range from 0.5 to more than 93 hours. Conclusion Calf is the right animal model for NIVAD implantation and there are some characteristics in its perioperative treatment. The establishment of this animal model may play important role for further improvement of NIVAD.