[Polymorphisms of RHCE gene among serologic RhD negative donors of Han population in Chinese Jiangsu area].

This study was purposed to investigate the RHCE gene polymorphisms in Chinese Jiangsu Han blood donors with and without RHD gene among serologic RhD negative population. PCR with sequence-specific primers (PCR-SSP) was used to detect RHCE genotype in 337 serologic RhD negative, RHD gene positive donors. The RHD gene-specific polymorphisms were also determined by PCR-SSP in these donors. The results showed that among 337 serologic RhD negative, RHD gene positive donor 20 were RHCE*C/C, 62 RHCE*C/c, 24 RHCE*c/c, 25 RHCE*E/e, and 81 RHCE*e/e; the allele frequencies for RHCE*C and RHCE*c were 0.4811 and 0.5189, respectively; and for RHCE*E and RHCE*e 0.1179 and 0.8821. Among 231 RHD gene negative donors, 3 were RHCE*C/C, 34 RHCE*C/c, 194 RHCE*c/c, 15 RHCE*E/e, 216 RHCE*e/e; the allele frequencies for RHCE*C and RHCE*c were 0.0866 and 0.9134, respectively; and allele frequencies for RHCE*E and RHCE*e were 0.0325 and 0.9675, respectively. It is concluded that the most prevalent allele of RHCE gene was RHCE*ccee in Chinese Han Jiangsu RHD gene negative population. There is statistical difference in RHCE genotype distribution among serologic RhD negative population with and without RHD gene.

A case of dispermic chimerism with normal phenotype identified during ABO blood grouping.

BACKGROUND: Human chimerism with normal phenotype derived from the fusion of two different zygotes is a rare phenomenon. We describe a case of a phenotypically normal 17-year-old diagnosed with dispermic chimerism during routine ABO blood grouping. METHODS: ABO grouping, ABO genotyping, karyotyping, human leukocyte antigen (HLA) typing, and short tandem repeat (STR) analysis were performed. RESULTS: Forward typing with anti-A and anti-B sera resulted in mixed-field agglutination of red blood cells. The mother and father were blood group O and AB, respectively. The proposita had O1, A201 and B alleles in the ABO locus; O1 was a maternal allele, while A201 and B were the paternal alleles. The proposita karyotype was 46,XX/46,XY. HLA typing revealed that the proposita had three alleles (46, 51, 54) at the HLA-B locus, with the additional allele of paternal origin. STR analysis identified three alleles for five of the 15 markers (D2S1338, TPOX, D8S1179, D19S433, and D21S11) analyzed in the proposita's blood- and skin fibroblast-derived DNA. The additional alleles of TPOX, D8S1179, and D21S11 were of paternal origin. CONCLUSIONS: The genetic findings suggest that this proposita was produced by dispermic fertilization of two identical haploid ova formed by parthenogenetic activation.

[Genetic polymorphisms of alcohol dehydrogenase-2 and aldehyde dehydrogenase-2 associated with the susceptibility on esophageal cancer].

OBJECTIVE: To evaluate the impact of alcohol dehydrogenase-2 (ADH2) and aldehyde dehydrogenase-2 (ALDH2) polymorphisms on the susceptibility of esophageal cancer. METHODS: A case-control study including 221 cases of esophageal cancer and 191 controls was carried out in Taixing city of Jiangsu province. ADH2 and ALDH2 genotypes were tested by PCR and denaturing high performance liquid chromatography (DHPLC). RESULTS: (1) Compared with ALDH2 G/G carriers, ALDH2 A/A (OR = 5.69, 95% CI: 2.51-12.18) and ALDH2 G/A (OR = 1.70, 95% CI: 1.08-2.68) carriers showed a significantly elevated risk of developing esophageal cancer, especially among alcohol drinkers with ALDH2 A/A (OR = 8.63, 95% CI: 2.07-35.95). (2) Statistical relation was not found between ADH2 genotypes and the risk of esophageal cancer, with regard to the status of alcohol consumption. (3) Whether subjects with whatever ADH2 genotype, ALDH2 G/A or A/A carriers was found to have significantly increased the risk of developing esophageal cancer, with ALDH2 A/A carriers appeared having higher esophageal cancer risk than those ALDH2 G/A carriers. (4)Compared those non-drinkers with both ALDH2 G/G and ADH2 A/A, drinkers with ALDH2 G/A or A/A and ADH2 G/A or G/G genotypes showed a significantly elevated risk of developing esophageal cancer (OR = 8.36, 95% CI: 2.98-23.46). CONCLUSION: These results revealed that it was not ADH2 but ALDH2 polymorphisms and drinking alcohol had a significant interaction with the development of esophageal cancer, suggesting that in order to help lowering the risk of esophageal cancer, individuals who are carrying ALDH2 A/A or G/A genotypes should be encouraged to reduce their consumption of alcohols.

[Coix lachryma jobi L varma-yuan induces apoptosis of jurkat cell line in acute T lymphoblast leukemia and its mechanism].

The aim of the present study was to investigate the anti-proliferation and pro-apoptosis effect of Coix lachrymajobi L varma-yuan on acute T lymphoblast leukemia cell line Jurkat cells and its mechanism. Jurkat cells were treated with Coix lachrymajobi L varma-yuan of various concentrations (0, 0.4, 0.8, 1.6 mg/ml) for 24h. The inhibitory ratio was measured by Cell Counting Kit-8. The effects of Coix lachrymajobi L varma-yuan on apoptosis of Jurkat cells were determined by Hoechst 33258, PI and Annexin V-FITC/PI double staining. The mitochondrial membrane potential was analyzed by JC-1 staining. The results demonstrated that Coix lachrymajobi L varma-yuan inhibited the proliferation of Jurkat cells, and induced chromatin condensation and fragmentation (characteristic of apoptosis) and loss of mitochondrial membrane potential. In conclusion, Coix lachrymajobi L varma-yuan can inhibit the cell proliferation and induce the apoptosis of Jurkat cells. These effects relate to loss of mitochondrial membrane potential. These results suggest that Coix lachrymajobi L varma-yuan may be of value in treating lymphoma.

Polymorphisms of alcohol dehydrogenase-2 and aldehyde dehydrogenase-2 and esophageal cancer risk in Southeast Chinese males.

AIM: To evaluate the impact of alcohol dehydrogenase-2 (ADH2) and aldehyde dehydrogenase-2 (ALDH2) polymorphisms on esophageal cancer susceptibility in Southeast Chinese males. METHODS: Two hundred and twenty-one esophageal cancer patients and 191 healthy controls from Taixing city in Jiangsu Province were enrolled in this study. ADH2 and ALDH2 genotypes were examined by polymerase chain reaction and denaturing high-performance liquid chromatography. Unconditional logistic regression was used to calculate the odds ratios (OR) and 95% confidence interval (CI). RESULTS: The ADH G allele carriers were more susceptible to esophageal cancer, but no association was found between ADH2 genotypes and risk of esophageal cancer when disregarding alcohol drinking status. Regardless of ADH2 genotype, ALDH2G/A or A/A carriers had significantly increased risk of developing esophageal cancer, with homozygous individuals showing higher esophageal cancer risk than those who were heterozygous. A significant interaction between ALDH2 and drinking was detected regarding esophageal cancer risk; the OR was 3.05 (95% CI: 1.49-6.25). Compared with non-drinkers carrying both ALDH2 G/G and ADH2 A/A, drinkers carrying both ALDH2 A allele and ADH2 G allele showed a significantly higher risk of developing esophageal cancer (OR = 8.36, 95% CI: 2.98-23.46). CONCLUSION: Both ADH2 G allele and ALDH2 A allele significantly increase the risk of esophageal cancer development in Southeast Chinese males. ALDH2 A allele significantly increases the risk of esophageal cancer development especially in alcohol drinkers. Alcohol drinkers carrying both ADH2 G allele and ALDH2 A allele have a higher risk of developing esophageal cancer.

[Research progress on biological characteristics and clinical application of endothelial progenitor cells--review].

Endothelial progenitor cells are precursors of endothelial cells, which are able to differentiate into mature endothelial cells. Studies are needed to increase more detailed understanding on the mechanisms of EPC-differentiation, survival, homing and distribution of the tissue. The human EPC has potential to be used as diagnostic and prognostic or therapeutic tools in the future. This review describes recent studies on the biological characteristics and clinical application of EPC, including immunophenotype and functional characteristics of EPCs, mobilization, release and differentiation of EPCs, EPC number and recruitment, clinical application of EPCs, and so on.

[Effect of highway transportation on the quality of red blood cells].

This study was purposed to investigate the effect of highway transportation on the quality of blood components so as to provide experimental basis to meet the needs of military operations. The transport condition was simulated by random vibration test. The red blood cells, leukocyte-reduced red blood cells, washed red blood cells were randomly vibrated (C Road) for 4 hours. Then, these blood components were stored in refrigerator for 15 days (4 degrees C). Six milliliters of blood were collected before vibration, after vibration, and at day 15 days of storage after vibration, respectively. The suspension was isolated. The free hemoglobin (FHb), routine hematological parameters, and biochemical indexes were determined. The results showed that FHb, lactate dehydrogenase (LDH), K(+) of red blood cells and leukocyte-reduced red blood cells did not significantly change after vibration and storage. However, FHb, LDH and K(+) of washed red blood cells increased significantly after simulated transportation (p < 0.05). The levels of these parameters at day 15 of storage after vibration were also significantly higher than those after vibration (p < 0.01). The changes of other hematological parameters were not significant in three blood components after vibration (C Road) and storage for 15 day. In conclusion, red blood cells and leukocyte-reduced red blood cells were qualified for clinic transfusion even after transportation within 4 hours for 15 day storage later, if they were kept in proper blood container and protected from damping. However, the washed red blood cells could not be used for clinic after similar transport in the military operations.

[Effects of triptolide on proliferation and apoptosis of Jurkat cell line in acute T lymphocytic leukemia].

The aim of this study was to investigate the anti-proliferation and pro-apoptosis of triptolide on Jurkat cell line in acute T lymphocytic leukemia. The Jurkat cells were treated with various concentrations of triptolide (0, 1, 2, 4, 8, 16 microg/L) for 12 hours. The inhibitory ratio was measured by Cell Counting Kit-8 assay. The effects of triptolide on apoptosis of Jurkat cells were determined by DNA fragmentation (DNA ladder), Hoechst 33258, PI and Annexin V-FITC/PI double staining. The results demonstrated that triptolide inhibited the proliferation of Jurket cells. The 50% inhibitory concentration (IC(50)) was 4.0 microg/L. Chromatin condensation in the cells treated with triptolide could be seen by light microscopy. DNA electrophoresis showed evidence of nuclear fragmentation (DNA ladder). The hypoploid (sub-G(1)) population was increased after treatment with triptolide. The translocation of phosphatidylserine at the outer surface of the cell plasma membrane could be induced by triptolide. After treatment with triptolide for 12 hours, the rates of apoptotic cells were significantly increased. Moreover, these pro-apoptosis effects were in time-dependent manner. It is concluded that triptolide can inhibit the proliferation and induce the apoptosis of Jurkat cells. This study provides experimental basis for clinical use of triptolide in leukemia therapy.

[Grape seed extract inhibits the growth of prostate cancer PC-3 cells].

OBJECTIVE: To investigate the inhibitory effect of grape seed extract (GSE) on the growth of prostate cancer PC-3 cells. METHODS: PC-3 cells were treated with GSE at the concentration of 100, 200 and 300 microg/ml for 24, 48 and 72 hours, respectively. The the inhibitory effect of GSE on the growth of the PC-3 cells and the kidney cells of SD rats was determined by MTT reduction assay, with primarily cultured kidney cells of 1-3 days old SD rats as the normal control. RESULTS: GSE significantly inhibited the growth of PC-3 cells in a concentration- and time-dependent manner, but had only a mild inhibitory effect on the kidney cells. CONCLUSION: GSE inhibits the growth of prostate cancer PC-3 cells and can be used as a new drug for the treatment of prostate cancer.

[Development of a novel protein carrier inducing immune response and binding DNA in gene therapy].

OBJECTIVE: To develop a novel protein carrier which can not only regulate the immune system but also deliver DNA into the tumor cell as an effective non-viral gene delivery system. METHODS: By using gene engineering techniques, we constructed a fusion protein containing the -COOH end of human hepatitis B virus core antigen (HBcAg), small home-to-cancer peptide ligand RGD and Glutathione S-transferase (GST), which was expressed in E. coli and purified by size exclusion chromatography and affinity chromatography. We labeled it with FITC to observe whether it could bind prostate cancer PC-3 cell lines, and meanwhile used it as a non-viral gene delivery carrier with the plasmid pEGFP-N1 that could express GFP in PC-3 cells. Furthermore, we observed the regulatory function of this fusion protein to the mouse immune system. RESULTS: The results of SDS-PAGE showed that the new protein carrier was obtained, which It could enter PC-3 cells with DNA in vitro and induce the mouse immune system to produce IgG1 and IgG2alpha simultaneously. CONCLUSION: The new protein carrier can be used as a target protein, especially in positive cells and the immune system. It promises to be a good novel carrier for the gene therapy of cancer.

Induction of apoptosis by recombinant soluble human TRAIL in Jurkat cells.

OBJECTIVE: To investigate the therapeutic potential of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells. METHODS: Expression of TRAIL receptors (DR4 and DR5) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Cytotoxic effects were determined by colony formation assay and a cell counting kit. The effects of recombinant TRAIL on apoptosis of Jurkat cells were determined by DNA fragmentation (DNA ladder) and PI staining. Changes in mitochondrial membrane potential were detected with JC-1 fluorescence. RESULTS: TRAIL inhibited the proliferation and induced internucleosomal DNA fragmentation (characteristic of apoptosis) and loss of mitochondrial membrane potential. CONCLUSION: Recombinant soluble TRAIL can be used as a therapy for cancer.

[Recent advances for research on anti-tumor effects of TNF-related apoptosis-inducing ligand].

Tumor necrosis factor (TNF)-related apoptosis--inducing ligand (TRAIL) is a member of TNF family. TRAIL selectively induces apoptosis in a wide variety of tumor cells, but not most normal cells through binding to its receptors. The ability of TRAIL to induce apoptosis in a large number of tumors has stimulated interest in TRAIL as a tumor therapeutic agent.

[Cloning, expression and purification of human TNF-related apoptosis-inducing ligand in Escherichia coli].

OBJECTIVE: To clone human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL, 114-281) cDNA and develop an inducible system for expression in E. coli. METHODS: The human TRAIL (114-281) cDNA was amplified with the total RNA from the human peripheral blood monocytes by RT-PCR. The cDNA was inserted into pMD T vector. The selected integrants were confirmed by PCR screen, digestion with restriction enzymes and DNA sequencing. Then, the insert of human TRAIL cDNA was subcloned into prokaryotic expression vector pET28a. The construction of prokaryotic expression vector was proofed correct by RT-PCR and digestive identification. The recombinant protein was induced by IPTG (isopropyl-beta-D-thiogalactopyranoside) . The sTRAIL inclusion bodies were refolded by dilution method. Refolded protein was purified with column chromatography. RESULTS: Agarose gel electrophoresis of product of RT-PCR revealed a band around 500bp, which was expected. The positive integrants were confirmed by PCR screen and digestion with restriction enzymes. The same band as RT-PCR product was showed by PCR screen and digestion with restriction enzymes. Then the selected integrants were confirmed by DNA sequencing. The sequence was checked in GenBank. The construction of prokaryotic expression vector pET 28a was proofed correct by RT-PCR and digestive identification. TRAIL protein was successfully induced by IPTG in E. coli BL21. The results also showed that the protein was expressed as inclusion bodies. After the sTRAIL inclusion bodies were solubilized and refolded, the protein expressed was purified with one band, which was about 20kD, analyzed by SDS-PAGE. CONCLUSION: These results suggested that the hsTRAIL was cloned, expressed and purified in the present study. Significant quantities of TRAIL produced by this method should allow further studies in determining the physiological significance and function of TRAIL in the future.