Adolescent ; Catheter Ablation ; instrumentation ; Equipment Failure ; Female ; Foreign Bodies ; etiology ; Humans ; Pulmonary Artery ; Silicones

Objective To explore the iodine nutrition level of people, prevalence of iodine deficiency disorders and the thyroid function of women of childbearing age in pasturing areas of Tibet. Methods Thirty families were selected respectively in pastoral Dangxiong county and agricultural Qushui county of Lhasa in 2009,drinking water and edible salt samples were collected to test iodine content; at least 50 people from each crowd of the following populations including children aged 8 - 10, women of childbearing age of 18 - 49 old and male adults aged 18 - 60 were randomly sampled and to measure their urinary iodine content and for thyroid palpation. Direct titrimetric method was used to test salt iodine(GB/T 13025.7-1999); As3+-Ce4+ oxidation reduction process to test water iodine (GB/T 5750.1-2006); As3+-Ce4+ catalytic spectrophotometry using ammonium persulfate digestion to test urine iodine(WS/T 107-2006), and goiter examination was based on Diagnostic and Classificatory Criteria of Endemic Goiter (WS 276-2007). Results The median of water iodine was 1.3 μg/L in pasturing area and 0.7 μg/L in agricultural areas, there was no statistical significant difference between them(Z =- 1.809, P ＞ 0.05).There was no iodized salt used in pastoral people, but iodized salt coverage rate was 90.0％(27/30) in agricultural residents. The median of urinary iodine among people of pasturing areas was 50.2 μg/L, lower than that of agricultural areas( 193.2 μg/L, Z =- 10.48, P ＜ 0.01 ). However, the goiter rate in pasturing area[1.0％(1/100)]was significantly lower than that of agricultural areas[18.0％(18/100) , x2 =16.8, P ＜ 0.01]. Serum level of FT4 and TT4 in pastoral population[(14.0 ± 2.0)pmol/L, (85.6 ± 17.5)nmol/L] was significantly lower than that of agricultural areas[(16.2 ± 6.3)pmol/L, (95.4 ± 21.1)nmoL/L, t =- 2.06, - 2.20, all P ＜ 0.05]. The thyroid dysfunction rate[5.9％ (2/34)]and subclinical hypothyroidism rate[2.9％ (1/34)]in pastoral population was significantly lower than that of agricultural areas[25.5％(12/47), 21.3％(10/47), x2 =5.328, 5.651, all P ＜ 0.05]. Conclusions Pastoral areas of iodine intake is significantly lower than the agricultural areas, urinary iodine levels reflect a serious iodine deficiency in pastoral people, but the blood biochemical and urinary iodine and goiter rate does not match,and shows hidden iodine hunger, which does not constitute a goiter epidemic.

OBJECTIVETo study the effect of lentiviral vector mediated herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) on graft- versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo- BMT) in mice.

METHODSDonor splenic lymphocytes from C57BL/6 which were infected by lentiviral vectors carrying HSV-TK were transplanted into 60Co gamma ray irradiated recipient mice with donor bone marrow cells. GCV 25 mg x kg(-1) x d(-1) was administered in 3 groups on day 0, +7, +12 respectively after transplant for 7 days by intraperitoneal injection. Survival time, severity of GVHD, incidence of GVHD, T lymphocytes immune reconstruction and of allogeneic chimerism ratio were detected after allo-BMT.

RESULTSThe average survival times for GCV 0 day, +7 day and +12 day group were (30. 10 +/- 5.21) d, (36.40 +/- 5.28) d and (28.20 +/- 4.82) d respectively, being significantly longer than that in the control group [(15.10 +/- 0.43) d] (P < 0.05). The 50 d-survival rate for TK/GCV + 7 day group was 60%. While for 0 day and +12 day group was 40% and 30% respectively. The incidence of grade III approximately IV GVHD in the control group was 100%, and the dead mice in experimental groups showed pathological changes of II approximately III GVHD. Long-term alive recipient mice only developed grade I approximately II GVHD after allo-BMT. The number of CD4+ lymphocytes in experimental groups was higher than that in control group (P <0.05), but CD8+ lymphocytes was lower on day +5, +10, +15 day (P <0.05). Allogeneic chimerism rate of recipient mice on +30 d was 100%.

CONCLUSIONSHSV-TK/GCV induced by the lentiviral vectors has a definite effect in prevention of GVHD after allo-BMT. GCV administrated from 7 days post-transplantation showed the best effects.

Animals ; Bone Marrow Transplantation ; immunology ; Ganciclovir ; pharmacology ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Lentivirus ; genetics ; Lymphocyte Transfusion ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; Transfection ; Transplantation, Homologous

Antibody against the angiotensin AT1 receptor (AT1-Ab) could disturb placental development. The placenta is the key organ between mother and fetus. Placental damage will seriously impair fetal growth and development in utero, leading to intrauterine growth restriction (IUGR). Based on the fetal origins of adult disease (FOAD) hypothesis, IUGR could increase a propensity to develop adult onset cardiovascular disease (CVD). The present study was designed to determine whether vascular function has changed in the adult offspring of AT1-Ab positive pregnant rats. Twenty four female rats (8-week-old, AT1-Ab negative) were randomly divided into two groups, immunized and vehicle groups. Immunized group received active immunization to establish AT1-Ab-positive model, while vehicle group was subjected to Freund's adjuvant without antigen. After 8 weeks of immunization, the antibody titers in sera from the female rats were detected by enzyme-linked immunosorbent assay (ELISA). Then all the female rats were mated with normal Wistar male rats and became pregnant. Immunized/vehicle group offspring rats (I offspring/V offspring) were raised to 40-week-old under standard chow feeding. Then the two groups' offspring rats were given a high-salt diet for 12 weeks (4% NaCl in chow feeding). Systolic blood pressure (SBP) was measured dynamically by noninvasive blood pressure system. The vascular ring experiment was performed to detect vascular function and reactivity. As detected by ELISA, the titers of antibody peaked at the 8th week (OD values: 2.75 ± 0.08 vs 0.33 ± 0.01, P < 0.01 vs vehicle group at the same time point). There was no significant difference of SBP between the two groups' offspring rats during the high-salt diet (P > 0.05). Isolated thoracic aortic rings of I offspring had significantly decreased constriction under norepinephrine treatment (P < 0.01 vs V offspring) and significantly decreased dilation under acetylcholine treatment (P < 0.05 vs V offspring). These results suggest that the offspring of AT1-Ab-positive pregnant rats are more susceptible to vascular functional abnormality while being fed high-salt diet.
Animals ; Antibodies ; blood ; Cardiovascular Diseases ; etiology ; physiopathology ; Disease Susceptibility ; Female ; Fetal Growth Retardation ; physiopathology ; Immunization ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; immunology ; Sodium Chloride, Dietary ; administration & dosage ; adverse effects

Objective To explore the status of iodine nutrition and iodine deficiency disorders in the pasturing areas and agricultural regions in Tibet. Methods 30 families were selected respectively in pastoral Dangxiong county and agricultural Qushui county of Lasa. Drinking water and edible salt were collected for testing the iodine contents. In each type of the following populations including children aged 8-10, women of child-bearing age and male adults, 50 subjects were randomly sampled to examine their urinary iodine contents. Among them, 50 children and 50 women were randomly selected for goiter examination by palpation. Results Water iodine content was less than 2μg/L, both in pasturing area and in agricultural areas. There was no iodized salt used in the families of pasturing areas, while 90% people consumed iodized salt in agricultural areas. The median of urinary iodine in pasturing area was 50.2 μg/L, significantly lower than that of agricultural area (193.2μg/L). However, the goiter rate of children and women in pasturing area was significantly lower than that in agricultural area. Conclusion Although iodine intake of populations in pasturing area of Tibet was severely deficient, there was no epidemic of Iodine Deficiency Disorders. This phenomenon noticed by the researchers deserved further investigation.

The study was purposed to prepare the recombinant lentiviral vector pTK161 and pTK162 carrying B-domain-deleted canine factor (BDDcFVIII) gene, and to investigate whether the canine FVIII (cVIII) can be expressed in vitro. The BDDcFVIII gene was ligated behind PUB and 2OH1 promotors to create lentiviral vectors pTK161 and pTK162. Meantime lentiviral vectors pTK161' and pTK161' were produced by cloning a green fluorescent protein (GFP) into pTK151 and pTK152, which was driven by PUB and 2OH1 promotors respectively. Vector supernatant were prepared by using transfer calcium phosphate mediated-cotransfection of 293T cells. The virus vector, DeltaNRF packaging-plasmid, and VSV-G envelope-plasmid was assayed by titers and cFVIII activity in cell culture supernatant after infection into 293T cells. pTK161, pTK162, pTK161' and pTK161' were identified by restriction enzyme analyzing. The results showed that the lentiviral vectors pTK161, pTK162, pTK161' and pTK161' were successfully constructed, and the titers of pTK161' and pTK161' reached to 1.54 x 10(6) U/ml and 2.83 x 10(6) U/ml; the activity of cFVIII could be detected at 24 hours after infection of 293T cells by pTK161 and pTK162, and achieved the highest level at 72 hours later. The higher level of cFVIII activity was achieved by transfected with pTK162 than that of pTK161 (p < 0.05), which closed to the cFVIII activity in normal dog plasma. 1/4 of the highest level could be detected 6 weeks later. It is concluded that the prepared HIV1-based lentiviral vectors can infect 293T cells to express cFVIII effectively. The results provide the basis for further studying HIV-1-based lentiviral vector gene therapy for hemophilia A.
Animals ; Dogs ; Factor VIII ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; HIV-1 ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

This study was aimed to construct a lentiviral vector carrying human coagulant factor VIII (FVIII) and to investigate its expression in 293T cells. B-domain-deleted factor VIII gene fragment (BDDhFVIIIcDNA) was obtained by enzyme digestion and cloned into lentiviral vector pXZ208 to establish the expression vector pXZ208-BDDhFVIII. Recombinant viral particles were prepared by cotransfection with packaging plasmid delta NRF and envelope plasmid VSV-G using calcium phosphate precipitation method. 293T cells were transfected by viral supernatant. Coagulant activity of FVIII, BDDhFVIIImRNA and genome integration were assayed by one-step method, RT-PCR and PCR after transfection. The results showed that 293T cells could be transfected by recombinant virus. The transfection rate of 293T was 59.57%. After transfection, the cells expressed FVIII efficiently. Detection confirmed that the activity of FVIII was 12%, 43% and 87% respectively at 24, 48 and 72 hours after infection. BDDhFVIII transcription was detected by RT-PCR from the infected cells. The gene integration in the targeted cells was also observed. It is concluded that the successfully constructed lentiviral vector is able to generate high level expression of human FVIII in 293T cells, which may provide a potential application of gene therapy to haemophilia A.
Cell Line ; Factor VIII ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVETo investigate the relationship between renal tubular cells transdifferentiation and chronic renal interstitial fibrosis induced by Fangchi Extract in rat.

METHODThe chronic renal interstitial fibrosis rat model was made by giving Radix Aristolochiae Fangchi extract (RAFE) and aristolichic acid (AA) respectively to rats through infusing stomach about 22 weeks discontinuously. Through immunnal histochemistry methods, investigating the expression of symbol proteins: Cytokine( CK) , alpha-Smooth muscle actin ( alpha-SMA) and Vimentin, and also the important fibrosis inducing factor-Transforming growth factor-beta (TGF-beta1 )on renal tubular cells.

RESULTIn RAFE and AA Groups, the expression of CK on renal tubular cells is declined comparing with the Control Group, and the enhanced expression of alpha-SMA and Vimentin can be observed on tubular cells. The expression of TGF-beta1 on renal tubular cells stronglhy increased, too.

CONCLUSIONPart of the renal tubular cells was transdifferentiated into myofibroblasts. Renal tubular cells may participate the occurance of chronic renal interstitial fibrosis, TGF-beta1 may accelerate the transdifferentiation of tubular cells.

Actins ; metabolism ; Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; isolation & purification ; toxicity ; Cell Transdifferentiation ; drug effects ; Cytokines ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Fibrosis ; Kidney Diseases ; chemically induced ; metabolism ; physiopathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Plant Extracts ; isolation & purification ; therapeutic use ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; Vimentin ; metabolism

OBJECTIVETo evaluate the effects of Tripterygium hypoglaucum (level) Hutch (THH) on cytokine production in acute graft-versus-host disease (aGVHD)mice and explore the mechanisms.

METHODS2 x 10(7) bone marrow cells mixed with 2 x 10(7) spleen cells from the same C57BL/6 mouse were transplanted into the myeloablative irradiated inbred BALB/c mouse to establish a aGVHD model. The experiments were designed as follows: control group (group A), CsA prophylaxis group (group B), THH prophylaxis group (group C), and combined THH with CsA prophylaxis group (group D). aGVHD was assessed by histologic changes of skin, liver and intestines. Chimerism was detected by H-2b molecular expression on recipient mice bone marrow cells with flow cytometer. Serum concentrations of IFN-gamma, IL-4 and IL-10 were determined by ELISA.

RESULTSThe serum concentrations of IFN-gamma in group B, C, D were significantly lower than that in group A, while those of IL-10 was significantly higher than that in group A (P < 0.05). There was no changes in concentration of IL-4 in all the groups (P > 0.05). The median survival time for group A was nine days, while that of group B, C, D each was more than 30 days being significantly longer than that of group A (P < 0.05). The recipient mice of group A displayed significant clinical symptoms of GVHD, and died within 20 days; whereas those of group B, C, D showed only ruffled fur and uplift posture. Histologic changes of liver and intestines in group B and C displayed a few lymphocytes infiltration while the histologic morphology of skin, liver and intestines in the survived mice of group D was normal. Allogeneic chimerism rates of group B, C, D at day 30 after allo-BMT were (99.18 +/- 0.58)%, (97.68 +/- 0.59)%, and (99.15 +/- 0.11)%, respectively.

CONCLUSIONTHH could regulate the production of cytokines and prevent aGVHD. THH and CsA at low dose combination showed synergic effect in preventing aGVHD.

Animals ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Graft vs Host Disease ; blood ; prevention & control ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Phytotherapy ; Tripterygium ; chemistry

OBJECTIVETo discuss the protective effects of pueraria compound on the cerebral ischemic injury.

METHODUsing the middle cerebral artery occlusion model (MCAO) in rats and cerebral ischemia-reperfusion models in gerbils and mice, we investigated the influence of pueraria compound on the brain water content and the infarct size, the cerebral apoplexy exponent, the contents of lactic acid (LA) and lipid peroxide (LPO), the activities of lactic dehydrogenase (LDH), glutathione peroxidase (GPx) and Na+ -K+ -ATPase.

RESULTPueraria compound obviously reduced the brain water content and the infrarct size in MCAO, improved motor abilities in the cerebral ischemia-reinfusion model of gerbils, decreased the contents of LA and LPO and increased the activities of LDH, GPx and Na+ -K+ -ATPase in cerebral ischemia-reinfusion model of mice.

CONCLUSIONPueraria compound has the function of antioxidation and protective effect on ischemic brain tissue.

Animals ; Antioxidants ; isolation & purification ; pharmacology ; Brain ; metabolism ; pathology ; Brain Ischemia ; metabolism ; pathology ; Drug Combinations ; Gerbillinae ; Glutathione Peroxidase ; metabolism ; Isoflavones ; isolation & purification ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; metabolism ; Lipid Peroxides ; metabolism ; Male ; Mice ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Soybeans ; chemistry