AlM: To explore the expression and significance of cysteine- rich 61 ( CCN1/Cyr61 ) in oxygen - induced retinal neovascularization ( RNV) of mice and study the inhibition effect of CCN1 specific siRNA on RNV.
METHODS:Two hundred healthy C57BL/6J mice were chosen and randomly divided into control group, hyperxia group, hyperxia control group and CCN1 treated group, with 50 mice in each group. The hyperxia control group was treated with vector plasmids by intravitreal injection. The CCN1 treated group received CCN1 siRNA recombinant plasmids by intravitreal injection. Adenosine diphosphate-ase ( ADPase) stained retina flat-mounts was performed to assess the retinal vascular profiles, HE staining was applied to count the number of vascular endothelial cell nuclei breaking through the internal limiting membrane, protein and mRNA level expression of CCN1 were measured by immunohistochemistry, Western blot and RT-PCR.
RESULTS: There were large nonperfusion area and a large number of vascular endothelial cell nuclei breaking through the internal limiting membrane ( 25. 25 ± 1. 26;23. 12 ± 1. 16 ) in the hyperxia group and the hyperxia control group. Regions of nonperfusion and vascular endothelial cell nuclei (8. 47±1. 15) were decreased in the CCN1 treated group compared to the hyperxia group and the hyperxia control group. Compared with the control group, there were high protein and mRNA expression of CCN1 in the hyperxia group and the hyperxia control group. The expression of CCN1 protein and mRNA were decreased in the CCN1 treated group compared with the hyperxia group and hyperxia control group (all P<0. 05).
CONCLUSlON: The abnormal expression of CCN1 has close relation with RNV. The development of RNV can be markedly inhibited by RNA interference targeting CCN1, which, we believe, will provide new molecular targets and a rationale for clinical developing new strategy for ROP therapy.

Objective To explore the diagnostic value of three-dimensional ultrasound parameters-vascularity index (VI) and the volume of kidney in chronic kidney disease (CKD). Methods A total of 80 CKD patients at different stages and 30 healthy adults were enrolled in the study. VI and the renal volume were calculated with three-dimensional ultrasound, while resistance index (RI), cortical thickness (CT) and the volume of kidney were calculated with two-dimensional ultrasound, and the correlation of those parameters were analyzed statistically. Results VI of CKD2 patients reduced significantly compared with the control group, and RI of CKD3 patients increased significantly, CT of CKD3 patients reduced significantly. The diagnostic sensitivity, specificity and diagnostic value of VI in patients with early and late CKD were all higher than thpse of RI. Compared with the control group, three-dimensional ultrasound of renal volume and two-dimensional ultrasound volume of CKD 1 patients increased significantly, three-dimensional ultrasound volume of CKD2 patients reduced significantly, and two-dimensional ultrasound volume of CKD4 patients reduced significantly. There was significant negative correlation among VI, three-dimensional ultrasound volume, CT, two-dimensional ultrasound volume and pathological change scoring (r=-0.67, -0.52, -0.43, -0.52). Conclusion VI and three-dimensional ultrasound volume can detect renal function damage more earlier and sensitively than RI and two-dimensional ultrasound volume.

Objective To explore the clinical efficacy of lnstaTrak3500 Plus with Fluoro Trak magnetic imaging guidance for spine surgery, Methods The construction and operation progress of the system were applied in thoracic-lumbar spine surgery, Results The electric-magnetic guided imaging system can improve success rate in thoracic-lumbar spine surgery,and all operative pedicle screw were inserted accurately with no related complications error. Conclusion The electric-magnetic imaging system can improve the accuracy of operations and guide mini-invasive surgery, which can significantly decrease X-ray irradiation dosage.

Traditional Chinese medicine (TCM) has definitely clinical effect in treating hyperlipidemia, but the action mechanism still need to be explored. Based on consulting Chinese Pharmacopoeia (2010), all the lipid-lowering Chinese patent medicines were analyzed by associated rules data mining method to explore high frequency herb pairs. The top three couplet medicines with high support degree were Puerariae Lobatae Radix-Crataegi Fructus, Salviae Miltiorrhizae Radix et Rhizoma-Crataegi Fructus, and Polygoni Multiflori Radix-Crataegi Fructus. The 20 main ingredients were selected from the herb pairs and docked with 3 key hyperlipidemia targets, namely 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), peroxisome proliferator activated receptor-α (PPAR-α ) and niemann-pick C1 like 1 (NPC1L1) to further discuss the molecular mechanism of the high frequency herb pairs, by using the docking program, LibDock. To construct evaluation rules for the ingredients of herb pairs, the root-mean-square deviation (RMSD) value between computed and initial complexes was first calculated to validate the fitness of LibDock models. Then, the key residues were also confirmed by analyzing the interactions of those 3 proteins and corresponding marketed drugs. The docking results showed that hyperin, puerarin, salvianolic acid A and polydatin can interact with two targets, and the other five compounds may be potent for at least one of the three targets. In this study, the multi-target effect of high frequency herb pairs for lipid-lowering was discussed on the molecular level, which can help further researching new multi-target anti-hyperlipidemia drug.
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Humans ; Hydroxymethylglutaryl CoA Reductases ; chemistry ; genetics ; metabolism ; Hyperlipidemias ; drug therapy ; enzymology ; genetics ; metabolism ; Hypolipidemic Agents ; chemistry ; metabolism ; Membrane Proteins ; chemistry ; genetics ; metabolism ; Molecular Docking Simulation ; PPAR alpha ; chemistry ; genetics ; metabolism ; Protein Binding ; Pueraria ; chemistry

BACKGROUND:Spinocerebel ar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners.
OBJECTIVE:To observe the effectiveness of neural differentiation of induced pluripotent stem cel lines induced by SCA3 and the stability of CAG copy number.
METHODS:Skin tissue of SCA3 patient was obtained clinical y, and specific skin flbroblasts were isolated and cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cel s. Patient-specific induced pluripotent stem cel s, normal person induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene.
RESULTS AND CONCLUSION:Induced pluripotent stem cel s that had identical genetic background to fibroblasts were successful y obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cel s. Each cel line could differentiate into neural stem cel s. The CAG number did not apparently alter before and after reprogramming as wel as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cel s derived from SCA3 into neural stem cel s was lower than that of normal person-derived induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10). These findings demonstrate that reprogramming can successful y establish human induced pluripotent stem cel s, and induced the differentiation of above cel s into neural stem cel s. In the whole process, CAG number did not obviously alter, which was consistent with body cel s of patients.

Lactobacillus casei was selected as an antigen delivery vehicle for the development of oral vaccine to express recombinant LTB and porcine parvovirus (PPV) VP2 protein. The fusion protein gene encoding PPV VP2 protein and LTB, was cloned into the surface expression vector pPG, and then the recombinant expression vector pPG-VP2-LTB was electrotransformed into Lactobacillus casei 393, generating recombinant strain pPG-VP2-LTB/L. casei 393. After induced by 2% Lactose in MRS broth, an about 78 kD protein was detected in the recombinant Lactobacillus casei by SDS-PAGE. The result of Western blot indicated that the protein possessed the antigenic specificity same as the native virus protein. The result of the whole bacteria cell ELISA indicated that the LTB protein was expressed at the same time. The results of indirect immunofluorescence test and immuno-gold electron microscopy showed that the interest protein was expressed on the surface of L. casei 393. The results provide potential for the development of lactic acid bacteria oral vaccine of PPV, which used LTB as mucosal adjuvant.

OBJECTIVETo study the clinical effect of segmental resection of the liver using Glissonean pedicle transection for primary liver cancer.

METHODSThe clinical data of 55 primary liver cancer patients admitted from January 2006 to October 2008 were analyzed retrospectively. Twenty-five of the patients underwent segmental resection of the liver by Glissonean pedicle transection (group A), and 30 underwent routine hepatectomy (group B). The positivity rate of the resection margin, micrometastasis in the hepatic parenchyma surrounding the lesions and postoperative recurrence rates were investigated.

RESULTSThe positivity rate of the resection margin was 4.0% in group A, significantly lower than that of group B. The number of histological micrometastasis was significantly higher in group A than in group B (16 vs 8). The median distance of histological micrometastasis was 6.8 mm (2.7-25.6 mm) in group A and 4.2 mm (2.4-9.0 mm) in group B. The one-year recurrence rate was significantly lower in group A than in group B (16% vs 26.7%).

CONCLUSIONGlissonean pedicle transection for segmental liver resection is a simpler procedure than routine hepatectomy for primary liver cancer and can reduce the number of histological micrometastasis and recurrence rate.

Carcinoma, Hepatocellular ; blood supply ; pathology ; surgery ; Female ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; blood supply ; pathology ; surgery ; Male ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; prevention & control ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.

METHODSThe pCI-neo-KH-ORF expression vector was constructed by DNA recombinant technique and was introduced into COS-7 cells and K562 cells by lipofectactin-mediated DNA transfection. Expression of KH-ORF mRNA was detected by RT-PCR. The effect of KH-ORF on cell cycle of COS-7 cells and K562 cells was evaluated by flow cytometry (FCM). Effect on cell proliferation of COS-7 cells was tested by MTT assay and that on K562 cells was analyzed by growth curves and LDH activity measurement.

RESULTS(1) KH-ORF mRNA was expressed both in COS-7 cells and K562 cells. (2) The cell cycle and cell proliferation of COS-7 cells were unaffected significantly. (3) The proportion of cells in S phase was increased in pCI-neo-KH-ORF-transfected K562 cells; and growth curves and LDH activity indicated enhanced cell proliferation.

CONCLUSIONKH gene may be a leukemia gene related to proliferation of K562 cells.

Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; physiology ; Genetic Vectors ; Humans ; K562 Cells ; L-Lactate Dehydrogenase ; metabolism ; Open Reading Frames ; genetics ; physiology ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; genetics ; S Phase ; Transfection

Objective To study the effect of ulinastatin on no reflow or slow flow in the infarct related artery in patient with acute ST-elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary interventional therapy (PCI).Methods 180 STEMI patients were divided into the control group (n=100) and the ulinastatin treatment group (n=80).The control group received conventional PCI treatment and the treatment group received conventional PCI treatment plus ulinastatin. The level of serum inflammatory factors IL-6,IL-10,superoxide converting enzyme,the infarct related coronary artery reperfusion TIMI flow grade (TFG) and myocardial perfusion grade (TMPG),the results of postoperative cardiac ultrasound examination and clinical outcomes were compared between the two groups.Results Compared with the control group,the level of IL-6 was decreased,while the levels of IL-10 and superoxide converting enzyme were increased significantly in the ulinastatin treatment group(P<0.05).The TFG and TMPG of the infarct related vessels were increased significantly in the ulinastatin treatment group. The left ventricular end diastolic diameter[(54.6 ± 5.2 mm vs. (50.4±4.6) mm,P=0.046)]and left ventricular ejection fraction [(58.4±10.2) % vs. (62.2±9.8) % P=0.048] showed statistical difference between the two groups.Compared to the control,the major cardiovascular event rate of the treatment group during hospitalization (1% vs. 5%, P=0.038), after one month (1.2% vs. 3%,P=0.046) and 6 months (3% vs 12%,P=0.018) were all significantly lower .There was no significant difference in mortality between the 2 groups.Conclusion Ulinastatin may lower the incidence of no flow and slow flow after emergency PCI,improve heart function and the lower the rates of MACE.

OBJECTIVETo investigate the expression of the miR-155 in human primary breast cancer and its clinical significance.

METHODSFrom February to June 2009, 45 pairs of specimens of human primary breast cancer and matched nontumor breast tissues were collected from the patients who received operation for breast cancer. Real-time polymerase chain reaction (RT-PCR) was used to detect the miR-155 expression in those specimens.

RESULTSThe stem-loop RT-PCR was sensitive and specific enough to detect the expression of the miR-155. The median relative expression of miR-155 was 0.360 in tumor samples, and it was 0.135 in matched nontumor breast tissues, the difference was statistically significant (P < 0.05). It's indicated that the up-regulation of miR-155 expression was associated with advanced TNM clinical stage (median 0.316, 0.358 and 0.417 respectively for stage I, II and III tumor, P = 0.002), lymph node metastasis (median 0.383 and 0.355 respectively for cases with positive and negative lymph nodes, P = 0.034), higher proliferation index [median 0.387 and 0.353 respectively for cases with high proliferation index (Ki67 > 10%) and low proliferation index (Ki67 ≤ 10%), P = 0.019], estrogen receptor-positive (0.367 and 0.318 respectively for cases with positive estrogen receptor and negative group, P = 0.041) and progesterone receptor-positive (0.398 and 0.335 respectively for cases with positive progesterone receptor and negative group, P = 0.029) in patients with breast cancer.

CONCLUSIONSThe expression of miR-155 is up-regulated in primary breast cancer, especially in patients with positive estrogen and progesterone receptor. miR-155 may play an important role in the proliferation, invasion and metastasis of human primary breast cancer, and it could be a indicator in the diagnosis and prognosis of primary breast cancer.

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Estrogen Receptor alpha ; metabolism ; Female ; Humans ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Receptors, Progesterone ; metabolism