Fatal poisoning by accidental ingestion of the "heartbreak grass" (Gelsemium elegans) verified by toxicological and medico-legal analyses.

We present a case of fatal poisoning from accidental ingestion of Gelsemium elegans (G. elegans), a rarely toxic plant. A 41-year-old man was found dead, at his home, 6 h after drinking homemade herbal liqueur during lunch.

[Differentially expressed genes in asthenospermia: a bioinformatics-based study].

Objective: To study the differentially expressed genes in asthenospermia to gain a deeper insight into the molecular mechanisms of the disease.

Methods: We analyzed the differentially expressed genes in asthenospermia using GATHER, PANTHER and ToppGene online bioinformatics tools.

Results: Our bioinformatics mining and analyses revealed that the differentially expressed genes in asthenospermia played important roles in the cellular protein and macromolecular metabolism, protein modification, cell death, cell apoptosis and apoptosis induction.

[Expressions of cysteine-rich secretory protein 2 in asthenospermia].

Objective: To investigate the mRNA and protein expression levels of cysteine-rich secretory protein 2 (CRISP2) in the sperm of asthenospermia patients, and explore their relationship with sperm motility and related molecular mechanism.

Methods: We collected 78 semen samples from adult male patients with asthenospermia and another 70 from healthy volunteers as controls. We extracted total RNA and total protein from the sperm following purification of the sperm by Percoll gradient centrifugation, and detected the relative expressions of CRISP2 mRNA and protein in the two groups by RT-PCR, SYBR Green real-time PCR and Western blot.

[Single- and two-layer gradient centrifugation in sperm separation: comparison and appraisal].

Objective: To appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation.

Methods: Twenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment.

[Literature-mining and bioinformatic analysis of androgen-independent prostate cancer-specific genes].

Objective: To compare the differences of the gene expressions in androgen-independent and androgen-dependent prostate cancer (ADPC), gain a deeper insight into the molecular mechanism of androgen-independent prostate cancer (AIPC), and find effective means for its clinical diagnosis and treatment.

Methods: Eats of genes highly-associated with prostate cancer were obtained by mining PubMed with the FACTA tool, and the specifically expressed genes in AIPC were analyzed with a set of bioinformatic tools including GATHER, PANTHER, STRING and ToppGene.

Results: A total of 128 genes specifically expressed in AIPC were identified, as compared with 23 that were specific to ADPC.

[Update of asthenospermia-related genes and proteins].

One of the most common causes of male infertility is asthenospermia, whose pathogenesis, however, is not yet clear. Recent researches have found that some genes (such as tektin-2, DNAI1, DNAH5, DNAH11, AKAP4, SEPT4 and Smcp) and proteins (such as sperm proteins ACTB, ANXA5, PRM1, PRM2 and SABP and seminal proteins Tf, PSA, PAP and Fractalkine) are associated with asthenospermia. The finding of these molecular markers has provided a base for the explanation of the molecular mechanism of asthenospermia, and these markers may become the diagnostic and therapeutic targets of the disease.

[Mining the specifically expressed genes in sperms based on the bioinformatics methods].

Objective: To analyze the specifically expressed genes in sperms for better understanding of the molecular characteristics of sperms.

Methods: The hybridization data the genes in the sperms, oocytes and 10 normal tissues were retrieved from the GEO database to identify the genes expressed specifically in sperms and the patterns of their regulation using such bioinformatic tools as GATHER, PANTHER and DAVID.

Results And Conclusions: Comparison of the spermatozoal gene expression profiles with those of the normal tissues identified 8998 differentially expressed probes, among which 25 genes were up-regulated by over 200 folds in the sperms.

[Mutation of MTCYB and MTATP6 is associated with asthenospermia].

Objective: To explore the correlation of the mutation of MTCYB and MTATP6 genes in sperm mitochondria with asthenospermia.

Methods: We extracted mtDNA from 80 semen samples of asthenospermia and 20 of normal sperm motility, amplified the MTCYB and MTATP6 genes by PCR, and analyzed their mutation by sequencing and BLAST matching.

Results: The deletion of both MTCYB and MTATP6 were detected in 20 of the 80 asthenospermia samples, MTCYB deletion in 16 and MTATP6 deletion in 4, accounting for 20% and 5% respectively.

[Research of the spermatozoal gene expression with gene microarrays].

Objective: To perform the detection of spermatozoal gene expression in order to accelerate the study of spermatozoal molecular biology.

Methods: To collect the healthy adults sperm and lymphocytes respectively, and then to extract the total RNAs from them by RNeasy mini kit (QIAGEN) or Trizol reagent. Corresponding cDNAs were produced, digested, ligated, finally labeled with Cy3 (sperm) and CyS (lymphocyte) in the course of RD amplifying reactions.

[Construction and preliminary identification of subtracted cDNA library of leukemia cell line K562].

Objective: Subtractive hybridization technology is a common method to screen and clone differentially expressed genes. This study was to construct subtracted cDNA library of leukemia cell line K562, and screen for differentially expressed genes.

Methods: cDNA fragments of K562 cells (tester), prepared by restriction display (RD), were subtracted with the Sau3A I-digested cDNA fragments of normal lymphocytes (driver).

[An initial examination of the spermatozoal gene expression profile].

Objective: To collect the normal spermatozoal gene expression sequence tags with the restriction display technique for constructing a microarray to understand spermatozoal gene expression profiles.

Methods: The total RNA extracted from normal human spermatozoa were reversely transcribed into cDNAs, which were digested by Sau3AI and linked to universal adapters (adapter 1) at both ends. According to the sequence of the adapter, a pair of primers (universal primers 1) was designed, followed by PCR with primers 1 and the PCR products were transferred into E.

[Analysis of gene expression patterns of leukemia K562 cells after cytochalasin B treatment].

Background & Objective: The advanced technique of DNA microarray makes it possible to monitor the expression of thousands of genes simultaneously in one hybridization experiment. This technique accelerates demonstration of anti-tumor drug mechanisms and discovery of new drug targets. This study was designed to investigate the differential gene expression of K562 cells after cytochalasin B treatment using cDNA microarray.

[Research advances on effect of arsenic trioxide on tumor].

Arsenic compounds are natural substances that have been used medically for more than 2,400 years in China. Since 1990s, Chinese physicians have began to use arsenic trioxide in treating the patients with acute promyelocytic leukemia (APL), and the results showed that arsenic trioxide was effective on APL patients. Further study of arsenic trioxide demonstrated that arsenic trioxide was safe and effective not only on patients with leukemia, but also on patients with many other kinds of malignant cancers.

[Proliferation and denucleation of K562 cells induced by cytochalasin B].

Objective: To observe the effect of cytochalasin B on the denucleation of K562 cells.

Methods: K562 cells were grown in RPMI 1640 medium supplemented with 10% calf serum and 4 microL cytochalasin B (CB). Denucleation was induced in the cultured cells by CB, and the cells were examined by phase contrast microscopy and Giemsa staining respectively.

[Detection of cell apoptosis by MTT assay].

Objective: To study the feasibility of using MTT assay to detect cell apoptosis.

Methods: K562 cells were grown in RPMI 1640 medium supplemented with 10% calf serum and 4 micromol/L arsenic trioxide. Apoptosis was induced in the cultured cells by As2O3, and the cells were detected with optical microscope, DNA gel electrophoresis and MTT staining respectively.

Preliminary study of DNA microarray of human placenta gene expression profile.

Objective: To prepare human placenta genechip for analyzing differential gene expressions.

Methods: The target gene amplified from Hybrid Hunter cDNA library was dotted onto the slides by the arrayer (PixSys 5500). Total RNA from K562 cells treated with or without arsenic trioxide was extracted, the mRNA purified and cDNA synthesized.

[Analysis with DNA chips of the changes of gene expressions in K562 cells in response to As2O3 treatment].

Objective: To investigate differential gene expression in apoptotic cells induced by As(2)O(3), and identify novel apoptosis-related genes.

Method: Apoptosis of K562 cells cultured in RPMI 1640 medium supplemented with 10 % calf serum was induced by As(2)O(3). Total RNA of the apoptotic and normal cells were then extracted, purified and subject to reverse transcription into first-strand cDNA, labeled with Cy3/Cy5.