Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

OBJECTIVETo purify caffeic acid tetramer (CAT) with macroporous resin on the basis of its fundamental physicochemical stability research.

METHODThe changes of CAT content were compared by HPLC method before and after the purification process, or while other conditions were altered.

RESULTLK001 was the best one among 7 kinds of macroporous resin in regard of purifying ability. The optimum absorbing technology was the solution concentration at 10 g x L(-1), pH at 4.5, and the flow rate at 3 BV x h(-1). The best eluting technology was 45% ethanol as eluting agent, pH at 5.0, eluting volume at 50 mL after applying super-purified water and 20% ethanol. The yield of product was 3. 6 percent, and the active compound CAT was 58 percent in the product.

CONCLUSIONMacroporous resin LK001 is effective in enriching CAT from the crude extracts, thus this method of purification is advisable.

Absorption ; Boraginaceae ; chemistry ; Caffeic Acids ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Hydrogen-Ion Concentration ; Light ; Oxygen ; chemistry ; Porosity ; Resins, Plant ; chemistry ; Temperature ; Water ; chemistry

OBJECTIVETo explore the better therapy in the treatment of ganglion.

METHODSNinety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups.

RESULTSAll of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01).

CONCLUSIONThe two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Ganglion Cysts ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism

Objective To study the effects of the serum Nesfatin-1 level change by type 2 diabetes in patients with Non-alcoholic fatty liver,and provides a reliable basis for its prysiological effects.Methods Selected 300 cases with type 2 diabetes mellitus in deoartment of endocrinology from January 2014 to January 2016 in Affiliated Hospital of Yan'an University as the object of this study.According to whether the patients were combined with non-alcoholic fatty liver,the patients were divided into T2DM with NAFLD group (150 cases) and T2DM without NAFLD group (150 cases).In addition,150 volunteers for healthy check-up in the hospital were selected as control group.Compared the serum Nesfatin-1 levels of three group patients,and monitored the insulin resistance index of two groups patients with type 2 diabetes.Results BMI,TG,ALT and INS of NGT(23.1±1.9,1.7±0.5,26.9±12.7 and 9.7±2.4,respectively) group and T2DM (22.1±2.5,2.0± 0.9,22.1 ± 10.5 and 11.2± 4.5,respectively) group were significantly lower than those of T2DM-NAFLD group (26.5 ± 3.8,3.0± 2.5,31.9 ± 11.5 and 14.2 ± 6.5;all P value were 0.00).FBG,HbAlc,LDL-C,HOMA-IR and Nesfatin-1 of T2DM group (8.4±3.1,8.8±2.7,3.4±1.0,4.5±2.9 and 6.9±3.0) and T2DM-NAFLD group (8.2±2.7,8.5± 1.9,3.7 ± 2.1,5.2 ± 2.7 and 5.2 ± 2.7) were significantly higher than those in serum NGT group (5.1 ± 0.5,5.4 ± 0.4,2.7 ±0.8,2.7±0.8 and 2.3± 0.7,all P value were 0.00).The serum Nesfatin-1 levels of T2DM patients and patients with T2DM-NAFLD had moderate correlation (r=-0.421,P＜0.05) by Pearson correlation analysis.BMI,TG and serum Nesfatin-1 levels were closely associated with the occurrence of fatty liver (r=-0.402 ～ 0.273;P=0.00 ～ 0.01).Conclusion Nesfatin-1 may be involved in type 2 diabetes mellitus and the occurrence of non-alcoholic fatty liver.

OBJECTIVETo investigate the clinicopathologic features of lymphoplasmacytic lymphoma (LPL) with Waldenström's macroglobulinemia (WM) and to evaluate the usefulness of immunophenotype analysis in diagnosis and differential diagnosis of the tumor.

METHODSA total of 40 cases of LPL with WM diagnosed according to the 2008 World Health Organization classification of tumors of hematopoietic and lymphoid tissues were analyzed using immunophenotype and follow-up information.

RESULTSThe mostly common initial clinical presentations were non-specific symptoms, such as fatigue, anemia and hemorrhage. Lymphadenopathy, splenomegaly and hepatomegaly were found in 42.5%, 20.0% and 12.5% of the patients respectively. The pattern of bone marrow involvement included mixed type (47.2%), diffuse type (41.7%) and interstitial type (11.1%). The nodal architecture was completely destroyed in one case and partially effaced with residual germinal centers and dilated sinuses in 8 cases. All of the neoplastic cells expressed CD20 and CD79a. Neoplastic plasma cells were positive for CD138 and CD79a. No cases expressed CD5. Four cases weakly expressed CD23. No significant prognosis related factors were identified in the survival analysis.

CONCLUSIONSLPL with WM is a rare indolent small B-cell lymphoma, which is commonly seen, in older male patients. The tumor frequently involves bone marrow and shows various clinical manifestations. Combination analyses of the bone marrow biopsy histology, immunophenotypic study and clinical data, especially the serum examination are important for the diagnosis of LPL with WM.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Bone Marrow ; metabolism ; pathology ; CD79 Antigens ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunoglobulin M ; blood ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphatic Metastasis ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology ; Neoplasm Invasiveness ; Survival Rate ; Syndecan-1 ; metabolism ; Waldenstrom Macroglobulinemia ; immunology ; metabolism ; pathology

OBJECTIVETo investigate the feasibility and efficacy on the outcome of patients with heart failure of integrated disease management program with heart failure clinic, patient education and telephone follow-up.

METHODSA total of 145 hospitalized patients with chronic heart failure and LVEF ≤ 45% or patients with LVEF > 45% and NT-proBNP > 1500 ng/L were divided into conventional group (n = 71) and interventional group (n = 74). Patients were followed for 10 to 12 months.

RESULTSBaseline clinical characteristics, LVEF and dose of evidence-based medicine were similar between the 2 groups. During follow-up, the NYHA functional class was higher in conventional group than interventional group (3.2 ± 0.5 vs 1.4 ± 0.5, P < 0.05), and the LVEF deteriorated in the conventional group and improved from 34% to 40%in the interventional group. The proportions of self-monitoring of weight, blood pressure and pulse rate in the interventional group were significantly higher than those of conventional group (P < 0.05). Among patients with systolic heart failure, 40% patients in the interventional group and 11% patients in the conventional group achieved the target doses of β-blockers (P < 0.05). Cardiovascular event rate of conventional group and interventional group is 91.5% and 27.0% respectively (P < 0.05).

CONCLUSIONIntegrated disease management program with heart failure clinic, patient education and telephone follow-up can improve patient compliance to heart failure treatment, improve cardiac function and reduce cardiovascular event rate.

Adult ; Aged ; Aged, 80 and over ; Ambulatory Care Facilities ; organization & administration ; Disease Management ; Female ; Heart Failure ; diagnosis ; therapy ; Humans ; Male ; Middle Aged ; Patient Compliance ; Prognosis ; Quality of Life ; Treatment Outcome

OBJECTIVETo investigate the effect of bone marrow stem cell transplantation (BMT) on the diaphragm muscles of mdx mice, a mouse model of Duchenne muscular dystrophy (DMD).

METHODSThe bone marrow-derived stem cells form male SD rats was transplanted through the tail vein into 18 female 8-week-old mdx mice, which were sacrificed at 4, 8 and 12 weeks after BMT (6 at each time point), respectively. The diaphragm muscles of the mice were subjected to HE staining, immunofluorescence detection of dystrophin, reverse transcription (RT)-PCR analysis of dystrophin mRNA transcripts and PCR analysis of Sry (sex-determining region on the Y chromosome) gene, with age-matched female C57 mice and untreated mdx mice as the controls.

RESULTSThe proportion of centrally nucleated fibers (CNF) in the diaphragm muscle of the recipient mdx mice was (15.58+/-0.91) %, (12.50+/-1.87) % and (10.17+/-1.17) % at 4, 8 and 12 weeks after BMT, respectively, significantly smaller than that of untreated mdx mice [(19.5+/-1.87) %], and the fibers after BMT showed less inflammatory infiltration. Compared with the untreated mice, the recipient mdx mice showed green fluorescence on significantly more diaphragm muscle cell membranes [with the proportion of dystrophin-positive fibers of (1.00+/-0.32) %, (6.00+/-1.05) % and (11.92+/-1.11) % at 4, 8, and 12 weeks after BMT]. RT-PCR of dystrophin mRNA also demonstrated significantly higher relative levels of dystrophin in the recipient mdx mice (0.19+/-0.05, 0.26+/-0.06 and 0.36+/-0.04 at 4, 8 and 12 weeks after BMT) than in untreated mdx mice, and Sry gene was present in the recipient mice.

CONCLUSIONBMT can partially restore dystrophin expression and ameliorate the pathology in the diaphragm muscles of mdx mice, and has great potential to produce general therapeutic effect in patients with DMD.

Animals ; Bone Marrow Transplantation ; methods ; Diaphragm ; metabolism ; pathology ; Dystrophin ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred mdx ; Muscular Dystrophy, Duchenne ; metabolism ; pathology ; surgery ; Rats ; Rats, Sprague-Dawley ; Transplantation, Heterologous

OBJECTIVETo construct the retroviral vector containing human micro-dystrophin gene and detect the expression of human micro-dystrophin in mdx mice bone marrow-derived mesenchymal stem cells (MSCs) after retrovirus infection.

METHODSRetroviral vector for micro-dystrophin gene was constructed and transferred into the packing cell PA317 mediated by Lipofectamine 2000. The retroviral supernatant containing the target genes were subsequently used to infect mdx mice MSCs. Micro-dystrophin expression was examined by methods of immunofluorescence staining and reverse transcriptase-polymerase chain reaction.

RESULTSMicro-dystrophin retroviral vector was successfully constructed and transferred into PA317 cells, and 48 h after infection with the recombinant retrovirus in mdx mice MSCs, 319 bp fragment could be detected by electrophoresis in the RT-PCR products. The red particles could be detected in some infected mdx mice MSCs with immunofluorescence staining. CONCLUSION mdx mice MSCs infected with retrovirus containing micro-dystrophin gene can express micro-dystrophin protein.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Dystrophin ; biosynthesis ; genetics ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred mdx ; Muscular Dystrophy, Animal ; metabolism ; Retroviridae Infections ; Transfection

OBJECTIVETo validate the value of the Oxford classification of IgA nephropathy in predicting the renal outcome in Chinese population.

METHODSRetrospective study was done in patients with IgA nephropathy. All slides were re-assessed according to the Oxford classification of IgA nephropathy. The primary end point is doubling serum creatinine, or a 50% reduction in estimated glomerular filtration rate (eGFR), or end-stage renal disease. Pathologic predictors for the progression to the end point were determined by univariate and multivariate Cox regression.

RESULTSTotally 533 patients were enrolled in the study. During the follow-up (median: 39 months; range: 12-263 months), 5.07% of the patients reached the end point. While tubular atrophy and interstitial fibrosis and arterial/ arteriolar lesion were associated with the endpoint in univariate analysis, only the T score was predictive of the renal outcome in multivariate Cox regression. Combination of the patho- logic lesions had no impact on renal outcome.

CONCLUSIONAccording to the Oxford classification of IgA nephropathy, the degree of tubulointerstitial fibrosis is the only feature independently predictive of renal outcome.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Glomerulonephritis, IGA ; classification ; pathology ; Humans ; Kidney ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult