Relationship between photosynthetic characteristics and environmental factors in leaves of Pueraria lobata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relationship between photosynthetic characteristics and environmental factors in leaves of P. lobata.</p><p><b>METHOD</b>Photosynthetic characteristics and environmental factors were measured by using CIRAS-2 portable photosynthesis system.</p><p><b>RESULT</b>The apparent quantum yield in leaves was 0.0173 micromol CO2 x micromol(-1) photon. The dark respiration rate was 2.9333 micromol x m(-2) x s(-1). The light compensation point of photosynthesis was 180 micromol x m(-2) x s(-1). The light saturation point was 1600 micromol x m(-2) x s(-1). The carboxylation efficiency was 0.0338 micromol x m(-2) x s(-1). The light respiration rate was 2.5 micromol x m(-2) x s(-1). The CO2 compensation point was 100 micromol x mol(-1), The CO2 saturation point was 1 600 micromol x mol(-1).</p><p><b>CONCLUSION</b>Photo flux density and air temperature are major environmental factors influencing diumal changes of net photosynthetic rate.</p>

Expression of liver-type fatty acid-binding protein and vascular endothelial growth factor and their correlation in human hepatocellular carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of liver-type fatty acid-binding protein (L-FABP) and vascular endothelial growth factor (VEGF) in human hepatocellular carcinoma (HCC) and its adjacent liver tissues, and investigate the correlation between the expressions of L-FABP and VEGF and their role in the occurrence and progression of HCC.</p><p><b>METHODS</b>Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical (IHC) staining were employed to examine the expression of L-FABP and VEGF in HCC and its adjacent liver tissues obtained from the surgical specimens of 61 HCC patients who underwent liver resections in West China hospital.</p><p><b>RESULTS</b>The results of RT-PCR showed that the expression level of L-FABP and VEGF in HCC was significantly higher than that in its adjacent liver tissues (L-FABP: 0.97-/+0.12, 0.83-/+0.14, t=5.21, P<0.05; VEGF: 0.92-/+0.11, 0.59-/+0.15, t=11.79, P<0.05). L-FABP tended to co-express with VEGF (P<0.05). IHC staining revealed that the expression of L-FABP and VEGF was mainly located in the cytoplasm, and the gray scale of L-FABP expression was significantly higher than that in the adjacent liver tissues (92.73-/+7.67, 82.83-/+6.90, t=7.44, P<0.05). The number of L-FABP- and VEGF-positive cells in HCC was significantly lower than that in the adjacent liver tissues (L-FABP: 92.18-/+4.44, 84.52-/+6.43, t=5.94, P<0.05; VEGF: 88.69-/+5.56, 77.64-/+5.93, t=8.72, P<0.05). Co-expression of L-FABP and VEGF observed in RT-PCR and also in IHC (P<0.05).</p><p><b>CONCLUSION</b>Both L-FABP and VEGF expressions are up-regulated in HCC. L-FABP gene may be involved in the carcinogenesis of human HCC. Expression of L-FABP is associated with VEGF expression, suggesting that L-FABP promotes the growth of blood vessels by taking up the fatty acids from the bloodstream, and both of them produce a marked effect on energy metabolism in HCC.</p>

Expression pattern of nitric oxide synthase in the retina of diabetic rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of nitric oxide synthase (NOS) in the retina of 8-week-old diabetic rats, and explore the potential molecular mechanisms for the role of NO in diabetic retinopathy (DR).</p><p><b>METHODS</b>Retinal gene expression profile of normal and 8-week-old diabetic rats was constructed with restriction fragment differential display polymerase chain reaction (RFDD-PCR). Bioinformatic analysis of the differentially expressed gene identified the genes coding for 3 subtypes of NOS, namely eNOS, nNOS and iNOS as the candidate genes related to DR, which was verified using semi-quantitative RT-PCR and immunohistochemistry.</p><p><b>RESULTS</b>The results of RFDD-PCR revealed down-regulated expression of eNOS and nNOS and up-regulated iNOS expression in diabetic rat retina. RT-PCR showed that the expression levels of eNOS and nNOS in diabetic rat retina were obviously lower than that in normal retina (0.23-/+0.03 vs 0.32-/+0.03 for eNOS, P<0.05; 0.25-/+0.02 vs 0.36-/+0.02 for nNOS, P<0.05), but the expression level of iNOS obviously higher (0.27-/+0.02 vs 0.20-/+0.03, P<0.05). Immunohistochemistry of healthy retina visualized eNOS-, nNOS- and iNOS-positive cells, all located in the inner nuclear layer (INL) and ganglion cell layer (GCL), and eNOS-positive cells were also found in vascular endothelium. In diabetic retina, the number of eNOS- and nNOS-positive cells was significantly lowered in comparison with normal rat retina (14.33-/+3.19 vs 22.13-/+3.60 for eNOS, P<0.05; 21.87-/+3.62 vs 34.40-/+7.09 for nNOS, P<0.05), but the number of iNOS-positive cells significantly increased (17.60-/+2.58 vs 11.73-/+2.70, P<0.05).</p><p><b>CONCLUSION</b>The alterations in eNOS, nNOS and iNOS expression are associated with the deuelopmant and progression of DR.</p>

Separation and determination of puerarin, daidzin and daidzein in stems and leaves of Pueraria thomsonii by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To separation and determine the contents of puerarin, daidzin and daidzein in the stems and the leaves of Pueraria thomsonii, and to provide scientific basis for developing and using of the stems and the leaves.</p><p><b>METHOD</b>A RP-HPLC method was applied with a Diamonsil C18 column (4.6 mm x 150 mm, 5 microm) by gradient elution using methanol-1% glacial acetic acid solution as the mobil phase. The flow rate was 1 mL x min(-1) and the detective wavelength was 250 nm, the column temperature was 25 degrees C.</p><p><b>RESULT</b>All of the three compounds showed good linearities (r >0.9995) and the recoveries were in the range of 99.0% - 101.6%. The contents of puerarin, daidzin and daidzein in the stems are higher than those in the leaves.</p><p><b>CONCLUSION</b>The method was accurate and could be used to contral the quality of the stems and leaves of P. thomsonii.</p>

Analysis of differential expression genes related to different metastasis potential of adenoid cystic carcinoma using restriction fragments differential display PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma.</p><p><b>METHODS</b>Restriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR).</p><p><b>RESULTS</b>Two gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24.</p><p><b>CONCLUSION</b>The construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.</p>

Islet formation and regeneration / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To explore the mechanisms of differentiation and development of pancreatic endocrine cells as well as pancreatic regeneration.</p><p><b>METHODS</b>Human embryonic pancreatic tissue at 7-14 weeks of gestation was collected. Diabetes mellitus rat model was induced with 65 mg/kg of streptozotocin. Insulin, glucagon, somatostatin, nestin, and cytokeratin 19 (CK19) of pancreatic tissues were observed by immunohistochemistry.</p><p><b>RESULTS</b>At 9 weeks of gestation, pancreatic epithelial cells began to co-express insulin, glucagon, somatostatin, and CK19 before migration. Islet cells gradually congregated along with the increase of aging, and at 14 weeks of gestation histological examination showed islet formation. At 12 weeks of gestation, nestin-positive cells could be seen in the pancreatic mesenchyme. During early embryogenesis, islet cells of pancreatic ducts co-expressed insulin, glucagon, and somatostatin. During pancreatic regeneration after damage, nestin expression of islet cells increased.</p><p><b>CONCLUSION</b>In the early stage of embryogenesis, islet cells of primary pancreatic ducts can be differentiated to multipotential endocrine cells before migration. During tissue regeneration, pancreatic stem cells may differentiate and proliferate to form pancreatic islet.</p>

Expression of CD34, CD14, CD10, CD31 and factor VIII on hematopoietic cells of yolk sac blood island, AGM region and liver of human embryo and fetus / 中华血液学杂志

<p><b>OBJECTIVES</b>To study the expression of CD34, CD31, CD14, CD10 and factor VIII on blood island of yolk sac (YS), PAS/aorta-germen-mesonephros (AGM) region and hepatic hematopoietic foci.</p><p><b>METHODS</b>Thirty-two cases of 3rd-12th weeks human embryo were obtained by drug abortion. Paraffin embedded sections with H.E staining and immunohistochemistry reaction (SABC) were performed.</p><p><b>RESULTS</b>YS blood island of 3rd-4th weeks of gestation was consisted of two types of cells. One was vascular endothelial cells located outside and the other hematopoietic cells inside the blood island. Both the two types of cells were CD10, CD14, CD31 and factor VIII positive. Hematopoietic cells were CD34 negative, and vascular endothelial cells were CD34 positive. On 32nd days of gestation, the hematopoietic cells migrated out of YS. On 4th week of gestation, CD34, CD14, CD10, CD31 and factor VIII positive cells appeared in the aorta, mesonephros and hepatic hematopoietic foci. By the 7th week, the number of positive hematopoietic cells reached the peak. In 11th-12th weeks, most cells in these regions were matured red blood cells and were negative for all the antibodies mentioned above excepting for CD34. During 4th-12th weeks, all endothelial cells in embryo were CD34 positive.</p><p><b>CONCLUSIONS</b>The hematopoietic cells and endothelial cells of YS blood island co-expressed CD10, CD14, CD31 and factor VIII. Endothelial cells were CD34 positive but hematopoietic cells were negative in YS blood island. The hematopoietic cells of aorta, mesonephros and hepatic hematopoietic foci expressed CD34, CD10, CD14 and factor VIII from 4th week to 7th week. Anti-CD34 antibody could label endothelial cells of every kinds vessels of embryo from 3rd to 12th weeks.</p>

Preliminary analysis of retinal gene expression profile of diabetic rat / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Establishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy.</p><p><b>METHODS</b>The whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential display-PCR (RFDD-PCR). Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach.</p><p><b>RESULTS</b>A total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, beta-arrestin, Phosducinìrod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mGluRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged.</p><p><b>CONCLUSION</b>These results prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro-dysfunctions of diabetic retina.</p>

Application of restriction fragment differential display-polymerase chain reaction in study on differential expression profiles of human diseases / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues.</p><p><b>METHODS</b>The tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed.</p><p><b>RESULTS</b>The expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments were found to relate with metastasis, differentiation, inflammation and so on.</p><p><b>CONCLUSION</b>RFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.</p>

Expression of VEGFA, VEGFC, angiopoietin-1, angiopoietin-2 and their receptors on development liver during gestation of weeks 3-12 of human embryo / 中国实验血液学杂志

The aim was to study the expression of VEGF-A, VEGF-C, angiopoietin-1, angiopoietin-2 and their receptors on development liver during gestation of weeks 3-12 of human embryo. Human embryo contingently aborted at 3-12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 microm serial sections were made. HE staining, immunohistochemistry method and light-microscope were employed. The results showed that at 4-5 weeks of development, liver was constituted by a few hepatic cords. Hematopoietic cell or blood cells were undetectable in the 4 week of gestation. A few cells which were larger, rounded and nucleared cells appeared and expressed VEGFA, flt-4 and Tie-2 proteins strongly in liver at 5 weeks of gestation. The number of these immuno-positive cells was highest in the 7th week and decreased at 11-12 weeks of gestation. These cells expressed flk-1 transiently in the 6th week. VEGF-C and flt-1 were expressed by hepatic cells from weeks 7 to 12 of gestation. The immuno-positive products were deposited in plasma of hepatic cells. Angiopoietin-1, angiopoietin-2 and Tie-2 were detectable on those cells which expressed VEGFA, flt-4 and Tie-2 from weeks 5 to 12 of gestation. The expression of angiopoietin-1 and angiopoietin-2 were weakly and Tie-2 was strongly. They were expressed weakly too by hepatic cells at 5 to 12 weeks of gestation. All factors and their receptors were undetectable on vascular endothelial cells at 4-12 weeks of gestation. It is concluded that the expression patterns of VEGF family on cells of liver are different before and after 7 weeks of gestation. The hematopoiesis in fetal liver may be related to development of hepatic cell.

Expression of VEGFA, VEGFC, angiopoietin-1, angiopoietin-2 and their receptors on yolk sac blood island, aorta-gonad-mesonephros (AGM) region of human embryo / 中国实验血液学杂志

The study was to investigate the expression of VEGFA, VEGFC, angiopoietin-1, angiopoietin-2 and their receptors on yolk sac blood island, AGM region during gestation of 3th-12th weeks of human embryo. Human embryo contingently aborted at 3 - 12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 micro m serial sections were made. HE and immunohistochemistry method (SABC) and light-microscope were employed. The results showed that VEGFA and its receptors flt1/flk-1, VEGFC and its receptor flt-4, angiopoietin-2 and its receptor tie-2 proteins were expressed strongly and angiopoietin-1 was weakly expressed by hematopoietic cells and vascular endothelial cells of blood island at 21 and 25 days of gestation. In the 4th week of gestation, immuno-positive reaction of these factors and their receptors appeared in the aorta and mesonephros deposited in larger, rounded and nucleated cells which represented hematopoietic cells. Up to 7th week, positive hematopoietic cells in the regions were much abundant. The number of positive cells decreased at 8th week. Up to 12th week, almost all blood cells were immuno-negative. VEGFA, flt-1, flt-4, angiopoietin-1, angiopoietin-2 and Tie-2 protein were expressed mainly by gonad at 6 - 8 weeks, but it did not express VEGFC and flk-1. The immuno-reaction of the factors and their receptors could not detected in vascular endothelial cells during 3-12th weeks of gestation. It is concluded that hematopoietic cells and endothelial cells in blood island of yolk sac, mesonephros and dorsal aorta co-expressed some factors and their receptors in relation to vasculogenesis and hematopoiesis. Intraembryonic hematopoiesis began in the 4th week of gestation.