OBJECTIVETo study the effects of 50 Hz electromagnetic fields (EMFs) on DNA of testicular cells and sperm chromatin structure in mice.

METHODSMice were exposed to 50 Hz, 0.2 mT or 6.4 mT electromagnetic fields for 4 weeks. DNA strand breakage in testicular cells was detected by single-cell gel electrophoresis assay. Sperm chromatin structure was analyzed by sperm chromatin structure assay with flow cytometry.

RESULTSAfter 50 Hz, 0.2 mT or 6.4 mT EMFs exposure, the percentage of cells with DNA migration in total testicular cells increased from the control level of 25.64% to 37.83% and 39.38% respectively. The relative length of comet tail and the percentage of DNA in comet tail respectively increased from the control levels of 13.06% +/- 12.38% and 1.52% +/- 3.25% to 17.86% +/- 14.60% and 2.32% +/- 4.26% after 0.2 mT exposure and to 17.88% +/- 13.71% and 2.35% +/- 3.87% after 6.4 mT exposure (P < 0.05). Exposure to EMFs had not induced significant changes in S.D.alphaT and XalphaT, but COMPalphaT (cells outside the main population of alpha t), the percentage of sperms with abnormal chromatin structure, increased in the two exposed groups.

CONCLUSION50 Hz EMFs may have the potential to induce DNA strand breakage in testicular cells and sperm chromatin condensation in mice.

Animals ; Chromatin ; radiation effects ; ultrastructure ; Comet Assay ; DNA ; analysis ; radiation effects ; DNA Damage ; Electromagnetic Fields ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Spermatozoa ; radiation effects ; ultrastructure ; Testis ; cytology ; radiation effects

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

OBJECTIVETo observe the effect of composite factors, like long-term high-salt & fat diet and alcohol abuse on blood viscosity and blood pressure in rats, and compare with a model induced by high molecular dextran, in order to build a chronic hyperviscosity aminal model which is similar to human hyperviscosity in clinic and lay a foundation for efficacy evaluation on traditional Chinese medicines.

METHODMale SD rats were randomly divided into the normal group, the high molecular dextran (HMD) group and the high salt & fat and alcohol (HSFA) group. The HMD group was given normal diet and water for 23 day and then 10% HMD through tail vein for 5 days. The HSFA group was fed with high salt and high fat diets every day and alcohol for 20 h x d(-1) for 13 weeks. After the modeling, whole blood viscosity and plasma viscosity were measured in the 5th, 8th and 11th week. Blood pressure was measured in the 5d, 7h, and 10th week. Red cell count (RBC) and hematocrit (HCT) were measured in the 11th week. PAgT, Fb, ET-1, NO, PGI, TXA2 contents of the normal group and the HSFA group were measured in the 13th week, and IECa21 content was measured with flow cytometry. Result: After the modeling, the HMD group was in good conditions with glossy hairs and active behaviors. The HSFA group was depressed with withered hairs and less activities. During the 5th-11th weeks, the HMD group and the HSFA group showed higher values in high and low shear whole blood viscosity (WBV) than the normal control group. The plasma viscosity (PV) of HMD rats was significantly increased only in the 5th week, and that of HSFA rats significantly increased in the 8"' and 11th week, particularly in the 11'h week. In the 111h week, the HSFA group showed significant increases in RBC and HCT. After the modeling, the blood pressure of HMD rats showed no significant changes, but the blood pressure of HSFA rats significantly increased during 7' and 101h weeks, particularly in the 10"' week. In the 13th week, PAgT, IECa2+, Fb, ET-1 of HSFA rats significantly increased, but with decreases in NO and PGI2.

CONCLUSIONLong-term high salt & fat and alcohol diets can cause abnormal blood viscosity in rats. WBV significantly increased since the 5th week in rats, and PV increased since the 8th week. The mechanism for increasing BV may be: (1) increases in RBC, HCT, and IECa2+, (2) PAgT increase, (3) Fb content increase, or (4) TXA2/PGI2, ET-1/NO imbalance. Although the modeling time with the method is longer than that with the HMD method, the model is more stable and moderate, and could lead to abnormal increases in WBV and PV; Whereas the HMD method only induced transient increase in plasma viscosity and abnormal increase in SBP. The model is more similar to traditional Chinese medicine syndromes and pathogenesis, with higher value for studies on efficacy of traditional Chinese medicines.

Alcoholism ; blood ; metabolism ; Animals ; Blood Pressure ; Blood Viscosity ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Ethanol ; adverse effects ; metabolism ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Chloride, Dietary ; adverse effects ; metabolism

Vascular endothelial growth factor-D (VEGF-D) and vascular endothelial growth factor receptor-2, -3 (VEGFR-2, -3) with their corresponding signaling pathway play significant roles in the development of the embryonic vascular system and pathological lymphangiogenesis. The study was aimed to express and purify the GST-VEGF-D fusion protein, and to explore the angiogenesis effect of VEGF-D. The total RNA was extracted from human fetal lung tissue, and the mature form of VEGF-D was expanded by polymerase chain reaction (PCR), then the plasmid pGEX-5X-1/VEGF-D was reconstructed and the GST-VEGF-D fusion protein expressed in transformed E.coli BL21-DE3. The results showed that the molecular mass of this fusion protein was 38 kD and compassed more than 15% of the total bacteria proteins. The fusion protein was recognized by anti-GST and anti-VEGF-D antibodies. The soluble GST-VEGF-D fusion protein could interact with VEGFR-3/Fc and was able to stimulate the proliferation of human erythroleukemia cell line (HEL) cells. The data of chick embryo chorioallantoic membrane (CAM) experiments indicated that GST-VEGF-D could induce the CAM angiogenesis. It is concluded that the GST-VEGF-D fusion protein with biological activity was successfully expressed, and which may provide an experimental model for the investigation of the VEGF-D-induced angiogenesis and lymphangiogenesis.
Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Humans ; Neovascularization, Physiologic ; drug effects ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Vascular Endothelial Growth Factor D ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo prepare monoclonal antibodies (mAbs) against enoyl-CoA hydratase 1 (ECH1).

METHODSNormal human liver tissues were homogenized, and the mitochondria were isolated by differential centrifugation. The total mitochondrial proteins were used to immunize BALB/c mice to prepare mAbs by routine hybridoma technique. The mAbs were characterized by ELISA, Western blotting and immunohistochemistry. The specificity of the antibody was identified by mass spectrometry (MS) following immunoprecipitation (IP) and confirmed by Uni-ZAP expression library screening.

RESULTSOne clone of the hybridoma BGB095 secreting specific mAb against ECH1 was obtained. The mAb was identified to belong to Ig subclass IgG1 and could be used in ELISA, Western blotting, immunohistochemistry, and immunoprecipitation.

CONCLUSIONA hybridoma cell line stably secreting specific mAb against ECH1 has been established. The specific mAb against ECH1 can be of great value for functional and distribution studies of ECH1.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; Antibody Specificity ; Blotting, Western ; Cell Line ; Enoyl-CoA Hydratase ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Liver ; cytology ; metabolism ; Mice ; Mice, Inbred BALB C ; Mitochondria ; metabolism

OBJECTIVETo study the expression of caveolin-1 in primary lung cancer and its relationship with microvessel density and clinicopathologic parameters.

METHODSImmunohistochemical study for caveolin-1 and CD34 was performed on paraffin sections of 154 cases of primary lung cancer and adjacent non-neoplastic lung parenchymal tissue, as well as 36 cases with nodal metastasis. Microvessel density was analyzed by CD34 immunostaining. Western blot assay was also employed in tumor and non-neoplastic lung tissues of the 50 cases (25 cases of pulmonary squamous cell carcinoma and 25 cases of pulmonary adenocarcinoma) with fresh specimens available.

RESULTSImmunohistochemical study showed that non-neoplastic bronchial and alveolar epithelium was positive for caveolin-1 (membranous and cytoplasmic). The expression rate of caveolin-1 in lung cancer was 59.1%, which was significantly lower than that in normal lung tissues (P < 0.01). Western blot assay confirmed that the expression of caveolin-1 in pulmonary squamous cell carcinoma and adenocarcinoma was lower than in surrounding non-neoplastic lung tissues (P < 0.01). Caveolin-1 expression in pulmonary small cell carcinoma (7.1%) was significantly lower than that in non-small cell carcinoma (64.3%) (P < 0.01). Within the group of non-small cell carcinoma, the expression of caveolin-1 was much higher in patients with lymph node metastasis (P = 0.005). The expression was also higher in stage III and IV than in stage I and II disease (P = 0.042).

CONCLUSIONSThe expression of caveolin-1 is lower in lung cancer tissues than that in non-small cell carcinoma, it is also significantly correlated with tumor stage and lymph node metastasis. Caveolin-1 may play some role in the progression of pulmonary non-small cell carcinoma.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caveolin 1 ; biosynthesis ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microvessels ; chemistry ; metabolism ; pathology ; Middle Aged ; Neoplasm Staging ; Small Cell Lung Carcinoma ; metabolism ; pathology

OBJECTIVETo control the seed quality market, a study on the seed determination practice in Polygala tenuifolia was carried out.

METHODBy studying the thousand grain weight moisture content, viability, genuineness, purity and germination percentage, some indices of seeds were fixed to the standards. The seed determination practice in P. tenuifolia was established.

RESULT AND CONCLUSIONThe practice could be utilized to control the seed quality of P. tenuifolia.

Germination ; Organ Size ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development ; Polygala ; anatomy & histology ; chemistry ; growth & development ; Quality Control ; Seedlings ; anatomy & histology ; Seeds ; anatomy & histology ; growth & development

OBJECTIVETo investigate the expression of connexin 43 and E-cadherin in lung cancer and to study the interaction between the two molecules.

METHODSThe expression and correlation of connexin 43 and E-cadherin were evaluated by immunohistochemistry (S-P method) in 85 samples of primary squamous cell carcinoma and adenocarcinoma of the lung. In addition, connexin 43 expression vector was transfected into the lung giant cell carcinoma cell line LH(7) followed by analyses of connexin 43 and E-cadherin expressions, the growth rates and cell cycle profiles of the transfected cells.

RESULTSComparing with the adjacent non-neoplastic lung tissue, expression of connexin 43 and E-cadherin was decreased in a correlative fashion in both squamous cell carcinomas and adenocarcinomas. Their expression reversely correlated to the degree of tumor cell differentiation, P-TNM stage, and status of lymph note metastasis. The expression of connexin 43 and E-cadherin increased significantly after transfection of connexin 43 expression vector into the LH(7) cells (P < 0.05). Both expressions were limited in the cytoplasm before or after the transfection. The proliferation rate of LH(7) cells was significantly decreased by connexin43 expression (P < 0.05), along with an increase of cell population at G(1) phase and a decrease of percentage of cells in S and G(2) phases (P < 0.05).

CONCLUSIONSSquamous cell carcinoma and adenocarcinoma of lung have a low level of connexin 43 and E-cadherin expression, which are correlated with the clinicopathologic features of the tumors. Transfection expression of connexin 43 gene induces an E-cadherin overexpression and an inhibition of LH(7) cell proliferation indicating the significant role of onnexin 43 in the regulation of cell proliferation.

Adult ; Aged ; Cadherins ; metabolism ; Cell Adhesion ; Cell Adhesion Molecules ; metabolism ; Cell Proliferation ; Connexin 43 ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Tumor Cells, Cultured

OBJECTIVETo establish a seed testing methods for Salvia miltiorrhiza.

METHODReferring to the International Seed Testing Rules made by ISTA and the Seed Testing for Crops (GB/T3543. 1-1995) issued by China.

RESULTThe seeds are selected by winnowing; the seed purity is about 50%-60%; 100 grain weight is used to determine the quality of the seed; the seed moisture content is determined by air drying, the drying hour is 3 h. Seed viability is tested by TFC method.

Chromosomes, Plant ; genetics ; Germination ; Quality Control ; Salvia miltiorrhiza ; chemistry ; genetics ; physiology ; Seeds ; chemistry ; genetics ; physiology

Mycoplasma hyopneumoniae (M .hyopneumoniae) and Mycoplasma hyorhinis (M .hyorhinis) infections are common in China .To investigate the prevalence of M .hyorhinis and M .hyopneumoniae in Jiangsu Province of China ,a mo-lecular epidemiological survey was conducted from 399 nasal swab samples of unvaccinated pigs using nested Polymerase Chain Reaction (nested PCR) .Nasal swab samples were collected from Jiangquhai porcine lean (JQHPL) strain pigs and other West-ern breeds .Clinical samples were taken from each pig and divided into different groups based on ages of pigs (7 ,14 ,21 ,28 , and 35 days) .Results indicated that the prevalence of M .hyorhinis was 70 .9% from different herds in Jiangsu Province in China ,while the prevalence of M .hyopneumoniae was 13 .5% .M .hyorhinis infection was more common in pigs for less than 5 weeks of age compared to M .hyopneumoniae infection .Co-infection was also observed in 30 samples (7 .5% ) in which both M .hyorhinis and M .hyopneumoniae were detected .The M .hyorhinis infection increased as the animals grew from 7 to 35 days .The M .hyopneumoniae infection did not change significantly as the pigs grew older .Significant difference of M .hyorhi-nis infection was observed between other Western breeds and JQHPL pigs (P<0 .001) .JQHPL pigs appear to be more sensi-tive to the M .hyorhinis infection as compared to the other Western breeds .However ,there is no obvious relationship between the breed type and M .hyopneumoniae infection (P>0 .05) .