Objective To evaluate the effects of 131 adrenoceptor anfisense on blood pressure and?1 adrenoceptor mRNA and protein levels in 2 kidney 1 clip(2K1C)rats.Method 2KIC hypertensive rots were produced by clipping renal artery of SD rats.Liposome/AS-ODNs 2.0 were tested intravenously in rats with 2KIC hypertension.Animals were divided into 5 groups(n=18 in each group):?1-AS-ODN group,?1-IN-ODN group,2K1C group,Sham group and SD group.Blood pressure was measured by tail-cuff method,the levels of myocardial?adreneceptor mRNA and protein were tested by RT-PCR and binding assay.Results On the basis of the magnitude and duration of hypotension,?1-AS-ODN decreased blood pressure by 39 mmHg at the most for 4 weeks.Compared with the 2KIC group,?1-AS-ODN did not significantly change the levels of myocardial?1 adrenoceptor mRNA but significantly decreased the levels of myocardial?1 adrenoceptor protein at 2,7,30 days (P

Rat osteoblasts were isolated from the 21-day fetal rat calvarias. The cells were grown in DMEM plus 10% FBS, and were treated for 24 h. With 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+ . Apoptosis of osteoblasts were measured by flow cytometry, electron microscopy and DNA fragmentation analyzed by gel electrophoresis. In addition, IP3 production and PKC activity were measured in order to show whether they are involved in apoptosis in osteoblast induced by zinc deficiency. The results showed that 10 μmol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells treated with 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+showed no apoptotic changes in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supplemented with Zn2 + simultaneously and the untreated cells. It can be inferred that apoptosis induced by zinc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.

Objective To establish a new murine model of experimental autoimmune myositis by immunizing with MYBPC2 protein. Methods The purified Myosin-binding protein C, fast type (MYBPC2) was emulsified with complete Freundˊs adjuvant, then C57BL/6 mice were immunized by multi-point subcutaneous injection (0, 7 days), and intraperitoneal injection of pertussis toxin 2 μg simultaneously. The pathological changes of mice with different immunizing dose at the preconceived time were ex-plored. Mean-while, mice were immunized with 600 μg each time, and the muscle endurance was tested on the 21th day. The expression of major histocompatibility complex (MHC) class-Ⅰ and the surface biomarkers of the inflammatory cells in muscle tissues were observed. Mann-Whitney U test was used for statistical analysis. Results ① With the increase of immunizing dosage, muscle damage and inflammation tended to be more serious. On the 21th and 28th day, muscle lesions were most significant. Muscle fiber degeneration and necrosis and inflammatory cell infiltration could be seen in the experimental group. ② Compared with the control group, muscle endurance of mice in the experimental group decreased significantly [(6.1 ±1.3) min versus (9.2±1.6) min, U=2.00, P=0.017]. The MHC class-Ⅰ on the muscle fiber surface of the experimental group was positive, scattered infiltration of CD4 +, CD8+ T ly-mphocytes and CD68 + macrophages between muscle fibers and around the vascular areas could be observed, and CD20+B lymphocytes mainly distributed in the area around the blood vessels, nevertheless rarely seen between muscle fibers. Conclusion Exper-imental autoimmune myositis models of mice have been successfully induced by immunizing with MYBPC2 in China for the first time, and similar clinical and pathological features of human polymyositis could be observed. This new model can be used for studying the pathogenesis of autoimmune myositis.

It has gradually become a consensus in the industry that the traditional Chinese medicine gypsum should be decocted first, but the understanding of decocting method is not completely unified in the works of doctors since ancient times, and there are occasional disputes about whether it is necessary to decocting first. In this study, the phase determination, physical and chemical characterization, qualitative and quantitative analysis of inorganic and organic components of the decoctions of herbal pairs and the whole prescription Maxingshigan decoction with gypsum as the center, and the pre-decoctions and co-decoctions of them were carried out to explore the scientific connotation of the pre-decoctions of gypsum. Results show that decoction phases were different between the co-decoctions and pre-decoctions of licorice-gypsum (Gancao-Shigao, GC-SG), ephedra-gypsum (Mahuang-Shigao, MH-SG) and almond-gypsum (Xingren-Shigao, XR-SG). The results of the micromorphology, particle size and zeta potential of herbal pairs and prescription (Quanfang, QF) showed that the supramolecular particles in pre-decoctions were smaller, more uniform and more stable than the co-decoctions. The results of organic components analysis showed that different cooking methods did not change the organic composition and content. ICP-OES results showed that the content of inorganic components in pre-decoctions was higher than in co-decoctions for the same boiling time of gypsum. The IR results showed that the pre-decoctions had stronger chemical functional group effect than the co-decoctions. To sum up, compared with the co-decoction, the pre-decoction of gypsum has different phase state and chemical composition interaction, and the difference of inorganic composition is an important material basis affecting the change of phase state compared with the co-decoction. It indicates that the material basis of traditional Chinese medicine decoction is indeed different whether gypsum is decocted first or not, which can provide a basis for the clinical application of decocted gypsum.

Objective To understand the related factors of rabies epidemic and provide the basic data for rabies control and prevention in China by statistic and retrospective analysis of rabies surveillance data in 2010.Methods We used descriptive epidemiology method and statistic analysis to analyze the epidemiological characteristics of rabies in 2010 of China.Results 2048 rabies cases were rabies cases were reported in 817 counties (districts) in 2010,which dropped 7.46％ compares to 2009.The incidences in children and elder people were high; farmers are main occupation of the cases,the male to female ratio of the cases was 2.44:1.Children and older people are higher acquired rabies than other age population.640 cases reported through national rabies sentinel surveillance system,87.50％ cases were caused by exposed to dogs,bite was the main exposure reason.The situation of deposing wounds was poor,and the use of vaccine was still low in individual cases,but in the rabies clinic cases under surveillance,the vaccine usage can reach 98％,the usage of immunoglobulin (RIG) or anti-serum for category Ⅲ exposure in either group cases was not high.Conclusion The epidemic of the rabies in 2010 was eased,Out-patient post-exposure prophylaxis was in good station,but there are still lots of problem existed: post-exposure prophylaxis of individual case was not desirable yet.

Evaluating toxicity and decoding the underlying mechanisms of active compounds are crucial for drug development.In this study,we present an innovative,integrated approach that combines air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI),time-of-flight secondary ion mass spectrometry(ToF-SIMS),and spatial metabolomics to comprehensively investi-gate the nephrotoxicity and underlying mechanisms of nitidine chloride(NC),a promising anti-tumor drug candidate.Our quantitive AFADESI-MSI analysis unveiled the region specific of accumulation of NC in the kidney,particularly within the inner cortex(IC)region,following single and repeated dose of NC.High spatial resolution ToF-SIMS analysis further allowed us to precisely map the localization of NC within the renal tubule.Employing spatial metabolomics based on AFADESI-MSI,we identified over 70 discriminating endogenous metabolites associated with chronic NC exposure.These findings suggest the renal tubule as the primary target of NC toxicity and implicate renal transporters(organic cation transporters,multidrug and toxin extrusion,and organic cation transporter 2(OCT2)),metabolic en-zymes(protein arginine N-methyltransferase(PRMT)and nitric oxide synthase),mitochondria,oxidative stress,and inflammation in NC-induced nephrotoxicity.This study offers novel insights into NC-induced renal damage,representing a crucial step towards devising strategies to mitigate renal damage caused by this compound.

AIMTo study the effects of FAK-ERK1/2 signaling pathway and FAK antisense oligodeoxynucleotides (ODNs) on vascular smooth muscle cell (SMC) migration and adhesion stimulated by fibronectin (FN).

METHODSMigration and adhesion of cultured SMCs were stimulated by different concentrations of FN, FAK, ERK1/2. And their phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense ODNs were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation, SMCs migration and adhesion were also measured by modifing Boyden Chamber and morphological enumeration, respectively.

RESULTSFAK were expressed when SMCs adhesion and migration were successfully simulated by FN (5, 10, 20, 40, 60 micrograms.mL-1), high contents of FAK and ERK1/2 phosphorylation were detected by 20 micrograms.mL-1 FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid. FAK and ERK1/2 phosphorylation were inhibited magnificently after FAK antisense ODNs transfection. Cell migration stimulated by FN 10, 20, 40 and 60 micrograms.mL-1 were reduced by 23.26%, 21.63%, 19.31% and 17.88% respectively (P < 0.05). SMCs adhesive spreading in 5-60 micrograms.mL-1 FN groups were reduced by 17.89%-27.67% (P < 0.05).

CONCLUSIONFAK-ERK1/2 mediated signal transduction play important roles in SMCs migration and adhesion stimulated by extracellular matrix. The process can be inhibited by FAK antisense ODNs effectively.

Animals ; Aorta ; cytology ; Cell Adhesion ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Fibronectins ; pharmacology ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Mitogen-Activated Protein Kinase 1 ; biosynthesis ; metabolism ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; biosynthesis ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; genetics ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transfection

Many studies have shown that G-CSF mobilized peripheral blood progenitor cell transplants (PBPCT) manifests faster recovery kinetics than conventional bone marrow transplants. This potential advantage of PBPCT still needs to be balanced against the risk of acute and chronic GVHD associating with the infusion of 10 - 15 fold higher donor lymphocyte number in unmanipulated allogeneic PBPCT than the marrow graft. To evaluate the effect of G-CSF primed bone marrow as a source of stem cells in the HLA-matched sibling transplantation, G-CSF primed with non-primed donor marrow in engraftment and incidence of GVHD for a homogenous group of patients with chronic myeloid leukemia (CML) were compared. Fifty patients with CML underwent bone marrow transplant, thirty-two donors (study group) were given G-CSF 3 - 4 micro g/kg per day for seven days prior to marrow harvest and eighteen donors (control group) had marrow harvest without G-CSF stimulation. Conditioning regimen consisted of total body irradiation and cyclophosphamide (CY), busulfan and CY, or busulfan, total body irradiation and CY. Both groups received same post-grafting GVHD prophylaxis and postgrafting G-CSF treatment. It was found that G-CSF primed donor marrow yielded with significantly higher number of total nucleated cells as well as CD34(+) cells and CFU-GM compared to non-G-CSF primed marrow (P = 0.001). The median engraftment time for absolute neutrophil (ANC > 0.5 x 10(9)/L) was day 15 (range 10 - 22) in the group of G-CSF primed vs day 21 in the non-primed donor group (P = 0.001). The median time for platelets (> 20 x 10(9)/L) was day 17.5 (range 13 - 28) in the group of G-CSF primed vs day 24 in non-primed group (P = 0.001). The incidence of acute GVHD grade II - IV in G-CSF primed donor group was surprisingly as low,as only two cases of thirty-two transplants (6.3%) with acute GVHD grade II limited to the skin. Whereas, five of eighteen patients (27.8%) in the control group developed acute GVHD grade II - IV (P = 0.032). G-CSF primed donors showed reduced CD4(+) and increased CD8(+) cells, resulting in a significant reduction of CD4(+)/CD8(+) ratio as compared with non-primed marrow. The total CD3(+) cell count kept unchanged in G-CSF primed donors. There were not significant differences in the incidence of the chronic GVHD (24% vs 33.3%), relapse rate (12.5% vs 11.1%) and overall survival rate (78.1% vs 66.7%, P = 0.32) during 6 - 50 months of follow-up. In conclusion, G-CSF primed donor marrow accelerates engraftment. Although G-CSF did not change the total CD3(+) cells in bone marrow, it altered the ratio of CD4(+) and CD8(+) cells and significantly reduced the incidence of acute GVHD.
Adolescent ; Adult ; Bone Marrow Transplantation ; Child ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Histocompatibility Testing ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Living Donors ; Male ; Transplantation, Homologous

OBJECTIVE: To investigate the effect and prognosis of stage I total hip replacement in the treatment of severe hip osteoarthritis with proximal femoral fracture. METHODS: From July 2014 to October 2017, 8 patients with severe end-stage hip disease and proximal femoral fracture were treated with stage I total hip replacement including 6 males and 2 females, aged 59 to 72 years old with an average age of 65 years old, involving 4 femoral head necrosis with proximal femoral fracture in the right side, 3 femoral head necrosis with proximal femoral fracture in the left side, and 1 left acetabular dysplasia with proximal femoral fracture in the left side. The average time from injury to operation was 7 days. Eight patients were treated with biologically elongated hip prosthesis. RESULTS: Eight patients with stage I total hip arthroplasty were followed up for 12 to 48 months with an average of 31 months. During the follow-up period, there was no loosening or subsidence of the prosthesis. Harris score increased from 33 points (22 to 42 points) preoperatively to 87 points(82 to 90 points) at the last follow-up. Among them, 3 cases were excellent and 5 cases were good. Abandoned abduction and walked 3 months after operation. X-ray films during 3-6 months after operation showed that fracture healing was good, hip pain and function were improved significantly, and the quality of life was greatly improved. CONCLUSIONS Phase I total hip arthroplasty for severe hip osteoarthritis patients with proximal femoral fracture has the advantages of shortening the treatment time, alleviating patients'pain, reducing hospitalization costs and good prognosis.
Aged ; Arthroplasty, Replacement, Hip ; Female ; Femoral Fractures ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Osteoarthritis, Hip ; Quality of Life ; Treatment Outcome

Background Bone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17β-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7. Methods After the treatment of MCF-7 cells with E2 at different concentrations (10-11 mol/L, 10-9 mol/L, 10-7 mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10-7 mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor α (Erα) on BMP-6 promoter in the presence of E2. Results E2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10-7 mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P<0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of Erα on 1/2 ERE response element in BMP-6 promoter was further validated. Conclusion Estrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor Erα binding on 1/2 ERE element in the BMP-6 promoter.