Triterpenoid saponins and phenylpropanoid glycoside from the roots of Ardisia crenata and their cytotoxic activities / 中国天然药物

Two new triterpenoid saponins, ardisicrenoside R and S (1 and 2), and one new phenylpropanoid glycoside, ardicrephenin (3), along with five known compounds (4-8), were isolated from roots of Ardisia crenata. Their structures were elucidated on the basis of NMR spectroscopic data and chemical methods. Compounds 2-7 were evaluated for their cytotoxic activities against A549, MCF-7, HepG2 and MDA-MB-231 cell lines by MTT assay. Ardicrenin (6) showed significant cytotoxicity, with IC

Two new steroidal saponins isolated from Anemarrhena asphodeloides / 中国天然药物

Two new steroidal saponins, named timosaponin P (1) and timosaponin Q (2), were isolated from the rhizome parts of Anemarrhena asphodeloides Bunge using various chromatographic methods. Their structures and absolute configurations were elucidated by a combination of spectroscopic and spectrometric data, including 1D, 2D NMR, HR-ESI-MS and ECD calculations, and this is the first time the absolute configuration of C-23 of steroidal saponin was confirmed by ECD calculations.

Steroidal saponins and their pharmacological activities in Solanum plants / 中国中药杂志

The genus Solanum which is the largest genus in Solanaceae consists of more than 2 000 species, most of them are distributed in tropics and subtropics areas, and only a small amount in temperate regions throughout the world. Steroidal saponins could be found in many species of the Solanum, and they are an important group of natural products exhbiting a number of potent beneficent properties, such as anti-tumor, antibacterial, antifungal, anti-inflammatory, antiviral, antioxidant, hypoglycemic, and antihyperlipidemic effects. As supplement, this paper gives a review of different structural categories of steroidal saponins and their pharmacological effects from the solanum plants over the past decade, and it is intended to provide a point of reference for further research on steroidal saponins in Solanum plants.

A260G and A386G single nucleotide polymorphisms of the DAZL gene are not correlated with male infertility in the Chinese population of Zhejiang Province / 中华男科学杂志

<p><b>OBJECTIVE</b>To assess the association of the A260G and A386G single nucleotide polymorphisms (SNP) of the DAZL gene with male infertility in the Chinese population of Zhejiang Province.</p><p><b>METHODS</b>We collected the peripheral blood samples from 317 idiopathic infertile males with azoospermia or oligozoospermia and 246 normal fertile men, and genotyped the polymorphic loci of the A260G and A386G polymorphisms of the DAZL gene using the SNaPshot technique.</p><p><b>RESULTS</b>The DAZL gene A260G was found genetically polymorphic in the Chinese population of Zhejiang Province, with the gene frequencies and their distribution consistent to the Hardy-Weinberg equilibrium. The frequencies of the AA, AG and GG genotypes of the A260G polymorphism were 92.3%, 7.3%, and 0.4% respectively in the normal controls and 94.3%, 5.7%, and 0% in the infertile patients, with no statistically significant differences between the two groups (P = 0.43, OR = 0.78, 95% CI 0.413-1.46). Heterozygosis (AG) of A386G was found in 1 of the control males but not in the infertile patients, while homozygosis (GG) of A386G was not observed in either group (P = 0.259, OR = 0.698, 59% CI: 0.374-1.306).</p><p><b>CONCLUSION</b>A260G and A386G SNPs of the DAZL gene are not associated with spermatogenic failure and neither represents a molecular marker for the genetic diagnosis of male infertility in the Chinese population of Zhejiang Province.</p>

Impact of male reproductive tract infection on semen quality / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association of male reproductive tract infection (RTI) with semen parameters and sperm DNA damage.</p><p><b>METHODS</b>We classified 1 084 males attending the infertility clinic into an RTI group (n = 300) and a non-RTI control group (n = 784). According to the WHO standards, we obtained routine semen parameters, detected sperm morphology, and determined the sperm DNA fragmentation index (DFI) by sperm chromatin structure assay.</p><p><b>RESULTS</b>There were statistically significant differences between the RTI and control groups in the semen volume ( [2.58 ± 1.20] vs [3.00 ± 2.10] ml), grade a + b sperm ([50.6 ± 17.2] vs [53.2 ± 15.8]%), grade d sperm ( [39. 8 ± 17.8] vs [36.5 ± 16.2]%), and total sperm count ([218.5 ± 185.0 ] vs [278.5 ± 375.5 ] x 10(6)/ejaculate) (all P < 0.05), but not in the males' age, sperm concentration or pH value (P > 0.05). The percentage of morphologically normal sperm was significantly lower ([3.46 ± 2.90] vs [4.61 ± 3.60%, P < 0.05) but the DFI was markedly higher in the RTI group than in the control ([19.4 ± 11.4] vs [15.2 ± 8.8]% , P < 0.01). The percentage of the cases with DFI > 30% was remarkably higher (13.0 vs 5.74% ) while that of the cases with DFI < 10% dramatically lower in the former than in the latter (16.0 vs 28.0%). The level of seminal plasma elastase was correlated negatively to sperm concentration, sperm count, and the percentage of morphologically normal sperm (P < 0.05) but positively to DFI and grade d sperm (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Male reproductive tract infection not only affects semen parameters and sperm morphology but also causes serious sperm DNA damage.</p>

FISH analysis of meiotic segregation results of the spermatozoa from male pericentric inversion carriers / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyze the meiotic segregation results of the spermatozoa from male pericentric inversion carriers by fluorescence in-situ hybridization (FISH).</p><p><b>METHODS</b>Using chemical depolymerization and multicolor FISH, we analyzed the meiotic segregation results of the spermatozoa from 4 male pericentric inversion carriers.</p><p><b>RESULTS</b>Of the 4 males studied, 46,XY,inv(9) (p11q12) was found in 2, 46,XY,inv(9) (p11q13) in 1 and 46,XY,inv(6) (p22q24) in the other; the lengths of the inverted segments represented 16.0, 16.0, 21.0 and 76.0% of the size of the whole chromosome involved; and the frequencies of recombinant sperm were 0.2, 0.4, 0.3 and 43.9%, del(p)/dup(q) accounting for 22.4% and del(q)/dup(p) 21.5%, respectively.</p><p><b>CONCLUSION</b>Males with pericentric inversion may produce spermatozoa with recombinant chromosomes and the rate of recombination varies principally according to the size proportion to the whole chromosome involved. The results of FISH analysis of chromosomal unbalanced spermatozoa can provide accurate personalized information on the genetic risk of fertility.</p>

Colon metastasis of chromophobe renal cell carcinoma with sarcomatoid change / 中华医学杂志(英文版)

We present a rare case of colonic metastasis of renal cell carcinoma (RCC) and review the literature. A 54-year-old male was referred to our hospital with a history of bloody stools and fever. A right kidney tumor measuring about 10 cm in diameter was found by abdominal computed tomography. Right radical nephrectomy and a right hemicolectomy with ileotransversostomy were performed. Pathological diagnosis was chromophobe RCC with sarcomotoid change involving the colon. Chromophobe RCC with sarcomotoid change is very rare.

Sperm DNA damage and sperm-nucleoprotein transition correlate to acrosin activity and seminal parameters / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters.</p><p><b>METHODS</b>We collected 535 semen samples, assessed sperm DNA damage by sperm chromatin dispersion test, and analyzed the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters according to the WHO criteria.</p><p><b>RESULTS</b>Statistically significant differences were observed in sperm DNA damage among sperm-nucleoprotein transition, acrosin activity, sperm concentration and the percentage of grade a + b sperm (P < 0.01). Sperm DNA damage was positively correlated with age, sperm-nucleoprotein transition, sperm concentration and the percentage of grade d sperm (P < 0.01 or P < 0.05), but negatively correlated with acrosin activity (P < 0.001). Stepwise linear regression analysis demonstrated that age, sperm concentration, the percentage of grade d sperm, sperm-nucleoprotein transition and acrosin activity were independent variables related to the DNA fragmentation index (DFI). The abnormality rates of sperm-nucleoprotein transition, acrosin activity, sperm concentration and graded a + b sperm were significantly higher in the sperm DNA damage group (DFI > or = 30%) than in the normal control (DFI < 30%) (P < 0.01).</p><p><b>CONCLUSION</b>Sperm DNA damage is closely related with sperm-nucleoprotein transition, acrosin activity and seminal parameters, which may become another important independent parameter for the evaluation of sperm quality.</p>

ANGPTL4 gene silencing by short-hairpin RNA inhibits the migration of human colorectal cancer cell line HT29 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of ANGPTL4 gene silencing on the migration of human colon cancer cells in vitro.</p><p><b>METHODS</b>The expression of ANGPTL4 in human colorectal cancer cell lines was detected by semi-quantitative RT-PCR. Following stable transfection with a short-hairpin RNA (shRNA) targeting ANGPTL4 gene in HT29 cells, ANGPTL4 mRNA and protein expressions were detected by semi-quantitative RT-PCR and direct ELISA, respectively, and the changes in cell migration ability and cell morphology were observed with transwell and immunofluorescence assays.</p><p><b>RESULTS</b>ANGPTL4 was expressed in most of the colorectal cancer cell lines. Compared with the control groups, HT29 cells with shRNA-mediated ANGPTL4 gene silencing showed significantly decreased expression of ANGPTL4 mRNA and protein (P<0.05) and lowered cell migration ability possibly due to decreased pseudopodia formation.</p><p><b>CONCLUSION</b>ANGPTL4 was expressed in most colorectal cancer cell lines. Decreased ANGPTL4 gene expression can inhibit the cell migration and pseudopodia formation in HT29 cells.</p>

A study on bisulfite sequencing method for methylation status of imprinted genes in single human oocytes / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish and improve the method of bisulfite sequencing for methylation status of imprinted genes in single human oocytes.</p><p><b>METHODS</b>Single superovulated immature human oocyte was embedded into low melting point agarose, followed by bisulfite treatment and polymerase chain reaction (PCR) amplification of the H19 and MEST genes. The PCR products were then subjected to TA cloning and sequencing to determine the methylation status.</p><p><b>RESULTS</b>With the modified methods of embedding and bisulfite treatment, we achieved a high PCR success rate of 82.46%, with the somatic cell contamination rate as low as 7.14%. The sequencing results showed no non-CpG cytosine and exact conformity to the theoretical sequences.</p><p><b>CONCLUSION</b>The bisulfite sequencing method we used to determine the methylation status of imprinted genes at the single-cell level was highly efficient and reliable, which can serve as a foundation for the further study of the influences of human assisted reproductive technology on genomic imprinting.</p>

Overcoming acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand by Bcl-XL small interfering RNA in human colon cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the reversing effect of Bcl-XL small interfering RNA (siRNA) on the acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human colon cancer.</p><p><b>METHODS</b>Human colon cancer cells DLD1-TRAIL/R, with acquired resistance to TRAIL, were firstly transfected with Bcl-XL siRNA for 24 h followed by the treatment of TRAIL protein. The survival rate of DLD1-TRAIL/R cells was assessed by FACS analysis and cell number counting, respectively, and activation of its apoptotic signaling was evaluated by Western blot.</p><p><b>RESULTS</b>Bcl-XL siRNA effectively downregulated the expression of Bcl-XL protein and reversed the acquired resistance to TRAIL in DLD1-TRAIL/R cells. After combination treatment of Bcl-XL siRNA and TRAIL protein, the apoptotic rate of DLD1-TRAIL/R cells was more than 50% and survival rate was less than 40%, whereas there was no effect on the survival of DLD1-TRAIL/R cells after treatment with control treatment or TRAIL protein treatment alone (P < 0.05). Western blot analysis demonstrated that caspase-8, caspase-9, Bid, caspase-3, and poly (ADP-ribose) polymerase (PARP) were obviously activated after combination treatment with Bcl-XL siRNA and TRAIL protein, and the release of cytochrome C was also significantly increased.</p><p><b>CONCLUSION</b>Bcl-XL siRNA can effectively reverse the acquired resistance to TRAIL in human colon cancer cells, suggesting that it might be a new strategy for overcoming the resistance in cancer therapy.</p>

Meiotic segregation results of male reciprocal chromosome translocations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the meiotic segregation results of male reciprocal chromosome translocation by fluorescence in situ hybridization (FISH).</p><p><b>METHODS</b>Multi-color FISH using 3 combined probes located in any 3 chromosome segments on both sides of two breakpoints was performed on the de-condensed sperm head to analyze the sperm chromosomal contents and segregation patterns.</p><p><b>RESULTS</b>Four male reciprocal translocation carriers were included in the study, with the karyotypes of 46, XY, t(2;18) (p16; q23); 46, XY, t(4;6) (q34;q21); 46, XY, t(8;13) (q23;q21) and 46, XY, t(4;5) (4q31;5q13), respectively. The results showed that 4 carriers had different proportions of various segregated spermatozoa. The spermatozoa of alternate, adjacent-1, adjacent-2, 3:1, non-disjunction in meiosis II, and 4:0 or diploidy accounted for 27.1%-49.4%, 26.9%-37.6%, 2.7%-15.7%, 8.6%-32.7%, 0.2%-1.9%, and 0.1%-0.4%, respectively.</p><p><b>CONCLUSION</b>For each-reciprocal translocation carrier seems to have a particular meiotic segregation results, FISH analysis on sperm head should be done for each carrier in order to provide an accurate genetic counseling.</p>

Tanscraniai doppler ultrasonography examination in diagnosis of the early cerebrovascular diseases in aged dia- betic patients / 中国实用内科杂志

Objective To evaluate the role of TCD examination in diagnosis of the early cerebrovascular diseases in aged diabetic patients.Methods 179 cases of aged type Ⅱ diabetic patients were divided into three groups(PDR,BDR and NDB)according to their retinopathy.TCD and retina examination were performed in all patients and the data be analysed. Results (1)The systolic peak flow velocity(Vp) of MCA、ACA、ICA and BA were significant increased in diabetic pa- tients than in normal control group(P

Activity of the TNF-related apoptosis-inducing ligand gene expressed from the hTERT promoter on colon cancer cell line HT-29 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the expression and activity of TNF-related apoptosis-inducing ligand (TRAIL) gene expressed from the hTERT promoter on colon cancer cell line HT-29.</p><p><b>METHODS</b>GFP/TRAIL gene expressed from the hTERT promoter was transfected into HT-29 with adenoviral vectors system, expression and apoptosis inducing ability of GFP/TRAIL protein were determined with fluorescence-activated cell sorting (FACS) method.</p><p><b>RESULTS</b>The expression of GFP gene was 31.4 % and 67.0 % with either hTERT promoter or CMV promoter in DLD1 cells; GFP/TRAIL gene was able to inhibit cell growth (74.2%) and induce apoptosis (25.8%) of HT-29 cells. There was significant difference between Ad/hTERT-gTRAIL and the other two control groups (PBS and Ad/CMV-GFP, P<0.05).</p><p><b>CONCLUSION</b>The GFP/TRAIL gene with hTERT promoter transfected by adenoviral vector was successfully expressed in HT-29 cell, which can both inhibit cell growth and induce apoptosis of colon cancer cell line HT-29.</p>

HLA-A site genotyping on single blastomeres is studied by nest-PCR-SSP method / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the accuracy and reliability of the nest-PCR-sequence specific primer(SSP) method in HLA-A site genotyping of single blastomeres retrieved from human pre-implantation embryos.</p><p><b>METHODS</b>By nest PCR on HLA-A exon 2, the success rate of first-round amplification was estimated for single blastomeres. Based on the first-round amplification, the HLA-A genotype of every single blastomeres was analyzed by commercially available PCR-SSP kits.</p><p><b>RESULTS</b>The amplification of HLA-A exon 2 were performed to 120 blasotmeres retrieved from in vitro fertilization(IVF) surplus embryos donated by 10 couples. The average success rate of family 1-5 and 6-10 was 78.2%(43/55) and 93.8%(61/65), respectively. And 86.7%(104/120) in total. Eighty blastomeres were further tested by nest-PCR-SSP, among which 11 blastomeres failed to HLA-A exon 2 amplification and then failed to genotyping while the other 69 blastomeres succeed in HLA-A exon 2 amplification and succeed in genotyping. Except for 6 blastomeres that were uncertain for allele lost because of parents' homozygosity, the left 63 blastomeres had accurate HLA genotyping. Among these 63 blastomeres, 59 blastomeres had genotypes confirmed from their parents(93.6%), 3 blastomeres lost one of parents' alleles(4.8%), and only one blastomere had two more than parents' alleles(1.6%).</p><p><b>CONCLUSION</b>The above research results indicated that based on the successful first round amplification of single blastomeres, nest-PCR-SSP strategy offers a convenient and reliable option for HLA genotyping on single blastomeres, which is a key process in pre-selecting HLA-identical sibling for allogeneic cord blood cell transplantation.</p>

Cloning and Analysis of Genes Correlated to Trehalose Biosynthesis from Micrococcus luteus / 微生物学通报

Genes related to trehalose biosynthesis from a bacterial strain Micrococcus luteus which can convert partially hydrolyzed starch into trehalose were cloned.Full sequence of gene (MtreY) encoding trehalose maltooligosyl trehalose synthase (MTSase) and partial sequence of gene (MtreZ) encoding maltooligosyl trehalose trehalohydrolase (MTHase) were got using PCR combined non-random shotgun method.Sequence analysis of MtreY predicts a 2370bp open reading frame encoding a protein of 790 amino acids with a predicted molecular weight of 86734 Da.Homologous analysis shows that this new gene has the same conservative motifs with ?-amylase family enzymes.The MtreY gene was expressed in E.coli, and the expression product has the anticipative enzyme activity.