Objective To observe the effects of fosinopril(FOS),a new generation angiotensin-converting enzyme inhibitor(ACEI),on protein and mRNA expression of transforming growth factor-?_1(TGF-?_1) of rat glomerular mesangial cell(GMC) induced by lipopolysaccharide(LPS);to demonstrate the preventive mechanism against glomerular sclerosis by applying FOS.Methods The cultured GMC in classic way were divided into 3 groups:control group;LPS group;LPS+FOS group.TGF-?_1 concentration in GMC supernatant fluid was detected by ELISA;TGF-?_1 mRNA expression was determined by semiquantitative real-time RT-PCR.Results LPS group was obviously higher than control groups in TGF-?_1 secretion and mRNA expression,while LPS+FOS group decreased distinctively in TGF-?_1 secretion and mRNA expression compared with LPS group.Conclusions FOS has obviously inhibited on TGF-?_1 expression of rat GMC both at protein level and mRNA level,which reveals that it may be an important mechanism by FOS on restraining the development of glomerulosclerosis.

Objective To evaluate the efficacy and safety of thromboprophylaxis with low molecular weight heparin after lumbar decompressive surgery.Methods Between January 2004 and April 2011,patients who had undergone lumbar decompressive surgery and had high or very high risk of venous thrombosis were selected.All patients received subcutaneous injection of low molecular weight heparin (Fraxiparine),starting at 6 hours after surgery with a half dose and subsequently once every 24 hours with full dose until discharge.When 24-hour drainage volume was less than 50 ml,the drainage tube was removed 2 hours prior to low molecular weight heparin administration.The occurrences of deep venous thrombosis (DVT),pulmonary embolism (PE),bleeding complications and side effects were recorded.Results Seventy eight patients were enrolled in the study.The average time of drug use was 8.5 days.No symptomatic DVT,PE and major bleeding events occurred.Drainage tube was placed in all patients except 3 patients with lumbar disc herniation.The mean total drainage volume was (319.5±218.5) ml,and the average time from operation to removal of drainage tube was (43.2±14.4) hours.Incision site ecchymosis occurred in 1 patient,incision bleeding in 1 patient,mild elevation in hepatic aminotransferase levels in 4 patients,and mild anaphylaxis in 1 patient.Conclusion It is effective and safe to prevent VTE with low molecular weight heparin for patients with high or very high risk of venous thrombosis after lumbar decompressive surgery.

OBJECTIVETo study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on apoptosis and cell cycle of mouse brain and liver cells.

METHODSMice were exposed to 50 Hz, 0.2 mT or 6.0 mT electromagnetic fields for 2 weeks. TUNEL and flow cytometric methods were used to analyze apoptosis and cell cycle of brain and liver cells.

RESULTSAfter exposure to 0.2 mT and 6.0 mT ELF EMFs for 2 weeks, apoptosis rates of brain cells [(5.60 +/- 1.47)% and (4.73 +/- 0.48)% respectively] were higher than that of control [(2.90 +/- 0.75)%], and apoptosis rates of liver cells [(4.19 +/- 2.08)% and (3.38 +/- 0.65)% respectively] were higher than that of control [(1.84 +/- 0.76)%]. G0/G1 cell percentage of brain cells [(80.21 +/- 1.68)% and (79.54 +/- 0.56)% respectively] were higher than that of control [(76.85 +/- 0.83)%], and those of liver cells [(79.42 +/- 1.80)% and (80.47 +/- 1.79)% respectively] were higher than that of control [(73.36 +/- 3.10)%]. The above differences were all statistically significant as P < 0.05. At the same time S and G2 + M cell percentage of brain and liver cells were significantly decreased.

CONCLUSIONExposure to 50 Hz EMFs may alter cell cycle and induce apoptosis of mouse brain and liver cells.

Animals ; Apoptosis ; radiation effects ; Brain ; cytology ; radiation effects ; Cell Cycle ; radiation effects ; Electromagnetic Fields ; Flow Cytometry ; In Situ Nick-End Labeling ; Liver ; cytology ; radiation effects ; Male ; Mice

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.
Apocynaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Flowers ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

OBJECTIVETo investigate the effects of extremely low frequency electromagnetic fields (ELF EMFs) on male reproduction in mice.

METHODS94 adult male mice were exposed to 50 Hz sinusoidal electromagnetic fields of 0.2, 3.2 or 6.4 mT for 2 weeks or 4 weeks. Testicular histology and weight, sperm amount, sperm motility and morphology were measured. The percentages of different ploidy cells and cell phases, and DNA content of testis cells were estimated by flow cytometry. The micronucleus rate of bone-marrow cell was also observed.

RESULTSThe testicular weight of the mice exposed to 6.4 mT for 4 weeks [(76.06 +/- 32.25) mg] was significantly lower than that of the control [(111.44 +/- 19.99) mg, P < 0.05]; no significant histopathological changes were observed on the testis in EMFs exposed mice;the sperm amount was decreased after EMFs exposure for 4 weeks, and those of the mice exposed to 0.2 mT and 6.4 mT for 4 weeks [(4.87 +/- 0.94) x 10(6)/ml and (4.30 +/- 1.89) x 10(6)/ml respectively] were significantly lower than that of the control [(6.67 +/- 0.70) x 10(6)/ml, P < 0.05]; the rates of sperm motility also showed a decline. After 0.2, 3.2 or 6.4 mT EMFs exposure for 2 weeks, the deformity rates of sperm [(7.416 +/- 3.352)%, (6.862 +/- 2.947)% and (8.112 +/- 4.615)% respectively] were significantly higher than that of the control [(4.098 +/- 2.028)%, P < 0.01]. Similarly, those of the mice exposed for 4 weeks [(10.267 +/- 3.836)%, (11.027 +/- 7.059)%, (8.814 +/- 3.678)% respectively] were higher than that of the control [(3.714 +/- 1.830)%]. After 6.4 mT exposure for 2 weeks, the percentages of 1C testis cells [(69.56 +/- 4.07)%] was significantly lower than that of the control [(73.45 +/- 3.10)%, P < 0.05]. There were not any remarkable changes in those of 2C, 4C cells. DNA content in different ploidy cells of the mice exposed to 6.4 mT was decreased. Moreover, the cell percentage in S phase was increased significantly (P < 0.01).

CONCLUSIONELF EMFs exposure may have some adverse effects on reproduction in mice.

Animals ; DNA ; metabolism ; Electromagnetic Fields ; Male ; Mice ; Random Allocation ; Reproduction ; radiation effects ; Sperm Count ; Sperm Motility ; radiation effects ; Spermatozoa ; cytology ; metabolism ; radiation effects ; Testis ; cytology ; radiation effects

It was recently discovered that a subset of osteoblasts functions as a key component of the hematopoietic stem cells (HSC) niche in vivo, controlling HSC self-renewal and multi-lineage differentiation. Disruption of Smad4 gene specifically in osteoblasts leads to a remarkable decrease of osteoblast number and endosteal surface area. In order to elucidate if the osteoblast loss has any effect on hematopoietic activity, the bone marrow (BM) and extramedullary hematopoiesis in the osteoblast-specific Smad4 knockout mice were systematically analyzed, the proportions of mature hematocytes in BM, liver and spleen were detected by flow cytometry, the hematopoietic progenitor number in different stages was measured by colong-forming assay, CFU-S and analysis of LSK cells. The results indicated that the conditional mutant mice demonstrated normal BM hematopoiesis without sign of extramedullary hematopoiesis. Furthermore, the proportion of hematopoietic progenitor cells was normal, while cell number/body weight of the conditional knockout mice increased. It is concluded that hematopoiesis is normally maintained in osteoblast-specific Smad4 knockout mice, and osteoblast loss does not of necessity result in the decrease in BM hematopoiesis.
Animals ; Bone Marrow Cells ; cytology ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; physiology ; Mice ; Mice, Knockout ; Osteoblasts ; cytology ; physiology ; Smad4 Protein ; genetics

OBJECTIVETo observe the effects of fosinopril (FOS) on proliferation and secretion of extracellular matrix of rat glomerular mesangial cell induced by LPS.

METHODSIn vitro culture method for glomerular mesangial cells (GMC) of rat was established and passages 3 - 10 of the cells were used in the experiment after identification. The experiment included the following 5 groups: control group (Ctrl), LPS group (LPS), high, medium and low dose FOS groups (FOS1, FOS2 and FOS3 groups, respectively). GMC proliferation was detected by methyl thiazolyl tetrazolium (MTT) incorporation method at 24 and 48 h; the changes of laminin (LN), fibronectin (FN) and ColIV protein secretion was detected by the enzyme-linked immunosorbent assay (ELISA). The changes of LNbeta(2) mRNA expression was detected by semi-quantitative real-time RT-PCR.

RESULTS(1) LPS could induce the mesangial cell proliferation, FOS inhibited this effect of proliferation induced by LPS. (2) Mesangial cells could secrete some extracellular matrix (ECM) protein in normal culture medium, mesangial cell secreted ECM protein was significantly higher in LPS group than that in Ctrl group (P < 0.01), but significantly lower in all FOS groups than that in LPS group (P < 0.01). (3) Mesangial cell could express LNbeta(2) mRNA in normal culture medium, LNbeta(2) mRNA expression was significantly higher in LPS group than that in Ctrl group at all time points, but was significantly lower in FOS group than that in LPS group.

CONCLUSIONSLPS could induce increased secretion of the ECM, including LN, FN, ColIV; FOS could inhibit the secretion of ECM in GMC in a dose-dependent manner at mRNA and protein levels.

Animals ; Cell Proliferation ; Cells, Cultured ; Extracellular Matrix Proteins ; secretion ; Fosinopril ; pharmacology ; Gene Expression Regulation ; Lipopolysaccharides ; Mesangial Cells ; drug effects ; metabolism ; Rats

Objective To study the effect of Shuangteng Bitong Cataplasm on TLR4, MyD88 and NF-κB and inflammatory cytokines in rats with collagen-induced arthritis; To discuss relevant mechanism of action. Methods A CIA model was made by subcutaneous injection of collagen emulsion from rat tail. Rats were randomly divided into normal group,model group,Shuangteng Bitong Cataplasm high-dose and low-dose groups.The contents of TNF-α, IL-1β and IL-6 in plasma were evaluated by ELISA, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues were evaluated by RT-PCR and Western blot. Results Compared with the model group, the contents of TNF-α, IL-1β and IL-6 in serum decreased remarkably in the Shuangteng Bitong Cataplasm low-dose and high-dose group, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues decreased remarkably. Conclusion Shuangteng Bitong Cataplasm can inhibit inflammatory reaction and regulate the expressions of relevant factors in synovial tissues to realize the anti-flammatory effects.

Objective:To investigate the influence of metastasis suppressor gene KAI1 on the proliferation,invasion and metastasis of endometrial carcinoma cell line AN3CA and HEC-1-B.Methods:The KAI1 cDNA was transfected into human endometrial carcinoma cells AN3CA and HEC-1-B via Lipofectamine 2000.The expression of KAI1 protein was ex- amined by Western blotting and flow cytometry before and after transfection.The proliferation ability of AN3CA and HEC- 1-B cells was observed by MTT assay and anchorage-independent growth assay.The changes of cell invasive ability were studied by transwell assays.Results:Stable expression of KAI1 protein was observed in AN3CA and HEC-1-B cells and on their surface after transfection with pcDNA3-KAI1 plasmid.Cells transfected with blank plasmid formed more colonies and had a larger size,with the colony forming rates being(54.2?3.1)% for AN3CA cells and(52.7?4.3)% for HEC- 1- B cells;the doubling time of AN3CA and HEC-1-B cells were 21.3 h and 20.1 h,respectively.Cells transfected with pcDNA3-KAI1 formed less colonies and had a smaller size,with the colony forming rates being(37.4?5.1)% for AN3CA cells and(32.1?3.7)% for HEC-1-B cells;the doubling time of AN3CA and HEC-1-B cells were 43.7h and 45.2 h,respectively.The cell proliferation abilities and colony-forming ability were significantly different between the two groups(P

Illegally paid blood donation was a risk factor for HIV acquisition exclusively in Henan and Hubei Provinces of China, and not in Shanghai. Nucleotide sequences in the gag and env genes of HIV-1 were compared between isolates from Henan and Shanghai regions of China to test whether an expected higher degree of a common source of infections from this unique blood donation transmission risk would be evident as decreased variation among Henan isolates in an exploratory cross-sectional analysis. Among 38 isolates studied, 23 of 23 (100%) from Henan and 8 of 15 (54%) from Shanghai were subtype B. In addition, fewer sequence differences were found in gp41 of subtype B isolates from Henan than from Shanghai isolates. Further studies with additional controls are therefore warranted to confirm the role of the degree of a common source of infections in differences in HIV variation across populations.