In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Objective To invesgate the effect of P-glycoprotein(PGP)inhibitor,verapamil,on electrobiological activity and seizure behavior in phenytoin-carbamazepine(PHT-CBZ)resistant rats.Methods The model of medically intractable epilepsy was established by kindling of amygdale. Verapamil was applied to PHT-CBZ resistant rats,followed by the observation on after discharge threshold (ADT),after discharge duration(ADD)and seizure activity.Results Compared with the control group, the ADT was higher in PHT-CBZ resistant rats peritoneally injected with verapamil((238.0?32.2)?A vs (177.0?23.3)?A,P

OBJECTIVETo investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).

METHODSThe MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.

RESULTSThe fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).

CONCLUSIONThe p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; Animals ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Transcription Factor RelA ; antagonists & inhibitors ; metabolism ; Transfection

The study was purposed to investigate the effects and mechanism of bone marrow-derived mesenchymal stem cells (MSCs) on graft-versus-host desease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The model of GVHD in rat had been established by allo-HSCT with donor derived T cells. The occurence of GVHD in recipients was observed in condition with or without donor derived MSC co-transplantation. Effects of MSCs on GVHD were analyzed by model rat survival rate and pathology. Proportions of CD4+CD25+ regulatory T cells were determined by using label spleen lymphocytes and thymocytes with double fluorescent-labeled antibodies and flow cytometry. The results showed that MSCs inhibited the lethal GVHD after HSC co-transplantation and increased the survival rate. The ratio of CD4/CD8 deceased in GVHD group in different levels, as compared with that in the experimental group. The proportion of CD4+CD25+ regulatory T cells of spleen lymphocytes was 31.55 +/- 7.58% and 20.90 +/- 1.90% in experimental and GVHD groups, respectively. Similarly, the proportion of CD4+CD25+ regulatory T cells of thymocytes was 93.20 +/- 2.69% and 57.17 +/- 6.79% in experimental and the GVHD groups, respectively. Meanwhile the proportion of CD4+CD25+ regulatory T cells was higher in experimental group than that in GVHD group. It is concluded that MSCs may prevent the lethal GVHD after allo-HSC co-transplantation and raise the survival rate of model rats by acting on the CD4+CD25+ regulatory T cells in vivo.
Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; adverse effects ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Graft vs Host Disease ; immunology ; Interleukin-2 Receptor alpha Subunit ; immunology ; Mesenchymal Stromal Cells ; immunology ; physiology ; Rats ; Rats, Inbred F344 ; Rats, Wistar ; T-Lymphocytes, Regulatory ; immunology

AIMTo study the chemical components from the bioactive extract of Herpetospermum pedunculosum, a Tibetan medicinal herb for liver diseases.

METHODSThe isolation and purification of this extract were conducted by means of silica gels column chromatography and preparative HPLC. The structures of the compounds were elucidated based on their physical and chemical features, and spectral data.

RESULTSTwo lignans were isolated from this extract. They were elucidated as herpetone (I), dehydrodiconiferyl alcohol (II).

CONCLUSIONCompound I is a new compound with the rare structure of sesqui-norlignan. Compound II is isolated from the genus of Herpetospermum for the first time.

Chromatography, Gel ; methods ; Chromatography, High Pressure Liquid ; Cucurbitaceae ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; Silica Gel ; Silicon Dioxide

OBJECTIVETo survey the incidence of acute febrile reaction in elderly patients receiving intravenous zoledronic acid for osteoporosis and identify the related factors.

METHODSThirty-eight elderly patients with osteoporosis were hospitalized and treated with intravenous infusion of 5 mg zoledronic acid in 2010. The incidence of acute fever reaction was observed in these patients , and the time of fever onset, duration, average maximum temperature, and antipyretic drug used were recorded. The patients with and without acute febrile reaction were compared for age, duration of osteoporosis, sex ratio, use of parathyroid hormone before zoledronic acid treatment, β-fragment of collagen breakdown, calcitonin, osteocalcin, serum calcium, and use of anti-osteoporosis drugs before the treatment.

RESULTSAcute fever reaction occurred in 12 (31.6%) of the patients. Two of the patients had fever on the day of zoledronic acid treatment, and the other patients developed fever after the first day of treatment, with a mean duration of 1 day and a maximum temperature of (38.5∓0.84) degrees celsius;. The fever was treated with a mean of 3.55∓1.21 pseudoephedrine tablets. The patients with fever showed significantly higher parathyroid hormone levels before treatment than those without fever (P<0.05); osteocalcin, calcitonin, β-fragment of collagen breakdown, or serum calcium showed no significant difference between the two groups.

CONCLUSIONAcute febrile reaction, often moderate and transient, is common in elderly patients receiving intravenous zoledronic acid for osteoporosis, and its occurrence is possibly associated with parathyroid hormone levels before the treatment.

Aged ; Aged, 80 and over ; Bone Density Conservation Agents ; administration & dosage ; adverse effects ; China ; epidemiology ; Diphosphonates ; administration & dosage ; adverse effects ; Female ; Fever ; chemically induced ; Humans ; Imidazoles ; administration & dosage ; adverse effects ; Incidence ; Infusions, Intravenous ; Male ; Osteoporosis ; drug therapy ; Parathyroid Hormone ; blood

OBJECTIVETo evaluate the influence of big Y chromosome on the outcomes of in vitro fertilization and embryo transfer.

METHODSData of 127 cycles of IVF/ICSI-ET, performed in our Reproductive Medicine Center from March 2001 to June 2003 were analyzed. The patients were divided into two groups according to the length of chromosome: Group A, 56 cycles with big Y chromosome

RESULTSNo significant difference was observed in the quality of embryos and in the and Group B, 71 cycles with normal karyotype. rates of fertilization, cleavage, clinical pregnancy, implantation, miscarriage, ectopic pregnancy, dead infant delivery, malformation,

CONCLUSIONBig Y chromosome has no significant influence on the baby boy delivery and baby girl delivery between the two groups. development of embryos and the outcome of pregnancy.

Chromosomes, Human, Y ; Embryo Implantation ; Female ; Humans ; Infertility, Male ; genetics ; therapy ; Male ; Pregnancy ; Pregnancy Rate ; Prospective Studies ; Sperm Injections, Intracytoplasmic

OBJECTIVETo study the clinical manifestations for norovirus gastroenteritis in infants and young children.

METHODSStool specimens were collected from infants and children with acute diarrhea who visited the affiliated Children's Hospital to Capital Institute of Pediatrics from January 2002 to December 2006. Enzyme-linked immunosorbent assay (ELISA) was used to detect human norovirus antigen in stool specimens and polyacrylamide gel electrophoresis (PAGE) was performed to detect rotavirus genome.

RESULTSOut of the 318 specimens under testing, 79 showed positive for norovirus antigen, with a positive rate of 24.8% (79/318). Among those positive specimens, 48(48/79, 60.8%) were detected in October to December, suggesting the seasonal preference of the virus. Most of the positive specimens (91.2%) were from those under 2 years of age. Rotavirus genome were detected from 16 out of 79 norovirus positive specimens (16/79, 20.3%), indicating those patients were co-infected by these two viruses. There was significant difference found in the severity of fever but not in the frequencies of diarrhea between rotavirus and norovirus co-infection group and noroviral infection group. Fourteen out of 79 norovirus positive patients were admitted to hospitals under the diagnosis other than gastroenteritis but started to develop symptoms of diarrhea between 1 to 11 days after hospitalization.

CONCLUSIONNorovirus seemed one of the most important pathogens for acute diarrhea among infants and young children and could cause nosocomial infectious gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; Child ; Child, Preschool ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Female ; Gastroenteritis ; diagnosis ; virology ; Genome, Viral ; genetics ; Humans ; Infant ; Male ; Norovirus ; classification ; genetics ; pathogenicity

Objective:To analyze therapeutic effect of amiodarone combined RAS inhibitors on hypertensive patients with atrial fibrillation(AF),and its influence on serum levels of uric acid(SUA)and silent information regulator 2-related enzymes 1(SIRT1).Methods:A total of 186 hypertensive patients with AF were selected from our hospital. They were randomly and equally divided into amiodarone group(group A),amiodarone + telmisartan group(A+T group)and amiodarone + ramipril group(A+ R group).Left atrial diameter(LAD),P wave dispersion(Pd),lev-els of high sensitive C reactive protein(hsCRP),brain natriuretic peptide(BNP)and SUA,SIRT1 mRNA and pro-tein expression and blood pressure variability(BPV)were observed and compared among three groups before and 45d after treatment.Results:Compared with before treatment,there were significant improvement in all index ex-cept SUA after treatment in three groups,P=0.001 all;compared with group A after treatment,there were signif-icant reductions in 24hSBPV[(0.112 ± 0.022)vs.(0.092 ± 0.020)vs.(0.091 ± 0.021)],24hDBPV[(0.143 ±0.031)vs.(0.130 ± 0.040)vs.(0.129 ± 0.039)],levels of hsCRP[(8.3 ± 0.3)mg/L vs.(5.3 ± 0.6)mg/L vs. (3.6 ± 0.4)mg/L],BNP[(64.9 ± 9.4)ng/dl vs.(61.1 ± 7.6)ng/dl vs.(58.2 ± 8.3)ng/dl]and SUA[(498.85 ± 89.54)μmol/L vs.(298.54 ± 56.12)μmol/L vs.(278.32 ± 54.09)μmol/L],SIRT1 mRNA[(2.20 ± 0.34)%vs.(1.87 ± 0.30)% vs.(1.76 ± 0.31)%]and protein[(29.54 ± 8.12)% vs.(26.31 ± 7.43)% vs.(23.21 ± 6.90)%]expression in A+T group and A+ R group,P<0.05 or <0.01. Compared with group A,there were sig-nificant rise in Pd[(39.3 ± 4.2)ms vs.(40.9 ± 4.1)ms,(41.2 ± 5.1)ms],and significant reduction in LAD [(37.8 ± 3.4)mm vs.(35.1 ± 4.6)mm,(35.7 ± 4.5)mm]in A+T group and A+R group,P<0.05 or <0.01. Conclusion:Amiodarone combined RAS inhibitors can significantly improve blood pressure variability,and reduce inflammatory factor,SUA and serum SIRT1 level in hypertensive patients with AF.

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P