Animals ; Apoptosis ; Cell Line, Tumor ; Down-Regulation ; Melanoma, Experimental ; pathology ; virology ; Mice ; Receptor, IGF Type 1 ; physiology ; Sendai virus ; pathogenicity

OBJECTIVETo explore PTEN gene expression and its clinical significance in acute leukemia.

METHODSThe expression levels of PTEN mRNA in 5 leukemia cell lines, 87 patients with acute leukemias (AL), including 59 acute myeloid leukemia (AML), 26 acute lymphoblastic leukemia (ALL), and 2 acute hybrid leukemia, 21 AL in complete remission (AL-CR), 31 chronic myelogenous leukemia (CML) and 14 normal controls were assayed by RT-PCR.

RESULTSPTEN mRNA was detected in K562 cell line, but not in Kasumi-1, HL-60, U937, Nalm-6 cell lines. The expression ratio of PTEN mRNA between CML (61.29%) and normal control (78.57%) had no statistical difference (P > 0.05). The expression ratios of PTEN mRNA in AL (18.39%) and AL-CR (42.86%) were significantly lower than that in normal control (P < 0.01 and P < 0.05, respectively), AL also has a lower expression ratio than that of AL-CR (P < 0.05). The decreased level of PTEN mRNA had a positive correlation with poor-prognostic factors (high white blood cell count of > or = 20 x 10(9)/L and chromosome abnormality).

CONCLUSIONThere is down-regulated expression of PTEN gene in AL. PTEN gene may play a role in leukemogenesis.

Cell Line, Tumor ; Humans ; Leukemia ; genetics ; metabolism ; PTEN Phosphohydrolase ; genetics ; metabolism ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective Studies on cardia-cancer caused by hereditary factors. Methods Case-control method was adopted,with information including name,sex,date of birth,date of death of all the Ⅰ,Ⅱ,Ⅲ relatives of the patients,diagnosis and the treatment collected. The hereditary probability of cardia cancer and the separation degree were calculated by Falconer and Li-Mentel-Gart. Results (1) Prevalence rates of cardia-cancer on relative Ⅰ,relative Ⅱ,relative Ⅲ of cardia-cancer patients appeared to be 0.54%,0.04%,and 0.05% respectively. Prevalence rates of upper-digestive-tract-cancer of relative Ⅰ,relative Ⅱ,relative Ⅲ of cardia-cancer patients showed as: 2.50%,0.36% and 0.13% respectively. Data showed that relative Ⅰ> relative Ⅱ> relative Ⅲ and family cluster existed in both males and females. (2) Cardia-cancer hereditary probability of the relative Ⅰ cardia-cancer probands was 11.71%,with males as 14.01% and females as 14.72%. The upper-digestive-tract-cancer hereditary probability of the relative Ⅰ cardia-cancer probands was 13.87%,with males as 11.49% and females as 23.08%,both below 25%,indicating this was a low hereditary cancer. (3) The upper-digestive-tract-cancer separation of the blood compatriots of cardia-cancer patients was 0.0452,with males as 0.0441 and females as 0.0507,both below 0.25,indicating the nature of a multi-gene but not single-gene hereditary way. Conclusion Hereditary factor is recognized as one of the high risk cardia cancer,but not the most risky factor causing the high morbidity of cardia cancer in Shanxi province.

Objective To investigate the effect of dihydromyricetin(DMY)on myocardial oxidative damage in exhaustive exercise mice.Methods C57BL/6 mice were divided into control group,model group,positive control group and low,medium and high dose experimental groups and with 10 mice in each group.Mice in control group and model group were intragastricated with distilled water;20,40 and 80 mg·kg-1 dihydromyricetin were given by gavage in low,medium and high dose experimental groups,while mice in positive control group were intragastricated with 100 mg·kg-1 Vitamin C once a day for 4 weeks.After administration,superoxide dismutase(SOD),malondialdehyde(MDA)and lactate dehydrogenase(LDH)were detected by the kit.The expression of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)protein were detected by Western blot.Results SOD levels in control group,model group and low,medium,high dose experimental groups and positive control group were(57.81±6.92),(26.85±2.74),(33.68±4.52),(39.74±3.95),(48.97±4.26)and(39.22±3.54)U·mg-1;MDA were(4.72±0.36),(10.48±1.68),(8.75±0.82),(6.43±0.71),(5.11±0.48)and(6.36±0.64)nmol·mg-1;LDH were(268.71±23.94),(726.58±81.26),(621.32±47.59),(479.12±50.24),(337.91±34.99)and(486.15±50.98)U·L-1;Nrf2 protein expression were 0.75±0.06,0.19±0.02,0.30±0.04,0.47±0.05,0.63±0.06 and 0.49±0.06;the protein expression of HO-1 were 0.83±0.08,0.27±0.05,0.39±0.04,0.52±0.03,0.77±0.07 and 0.55±0.06,respectively.There were statistically significant differences between control group and model group(all P＜0.05);there were statistically significant differences in the above indexes between model group and positive control group,low dose experimental group,medium dose experimental group,high dose experimental group(all P＜0.05).Conclusion Dihydromyricetin can delay myocardial oxidative injury in exhaustive exercise mice,which may be related to Nrf2/HO-1 pathway.

Objective To establish the activation of NF-κB in middle ear cholesteatoma, discuss the relationship of NF-κB and the gene expression of IL-6 and explore the pathogenesis of middle ear cholesteatoma. Methods Ten cases of cholesteatoma and 6 cases of normal external meatal skin were obtained from middle ear surgery. The NF-κB DNA binding activity and the mRNA level of IL-6 in these two kinds of tissues were detected by electrophoretic motility shift assay (EMSA) and Reverse transcription polymerase chain reaction(RT-PCR) respectively. The relationship of NF-κB DNA binding activity and the mRNA level of IL-6 were analyzed. Results The NF-κB DNA binding activities of cholesteatoma [(15.9±8.2) %] were higher than those in normal skin [(1.36 ± 0.94) %, t = 3.502, P < 0.05]. The expression of IL-6 mRNA was increased significantly in patients with cholesteatoma, as compared with that in the control specimens (t = 2.166 ,P < 0.05) and had a significant positive correlation with NF-κB DNA binding activity (r = 0.752, P < 0.05). Conclusions The II.,-6 mRNA expression in cholesteatoma is closely related with the activity of NF-κB. It is tempting to speculate that NF-κB play a key role in the activation of cytokine in cholesteatoma.

OBJECTIVETo set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.

METHODSFifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.

RESULTSIn the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively. In the exon 48/exon 19/SRY group, the amplification rates of exon 48, exon 19 and SRY in male were 92%, 90% and 94%, the amplification rates of exon 48, exon 19 in female were 94% and 92%, respectively.

CONCLUSIONThe technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene established in this study is highly sensitive, specific and reliable, and is suitable for PGD of deleted DMD with family history.

Dystrophin ; genetics ; Exons ; genetics ; Female ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Preimplantation Diagnosis ; methods ; Reproducibility of Results ; Sequence Deletion ; Sex Determination Processes

OBJECTIVETo achieve pregnancy with unaffected embryo using in vitro fertilization and embryo transfer (IVF-ET) and preimplantation genetic diagnosis(PGD) for the couples at risk of having children with beta-thalassemia.

METHODSA couple carrying different thalassemia mutations of codon 41/42 and codon IVS2 position 654 received standard IVF treatment and intracytoplasmic sperm injection, embryo biopsy, single cell polymerase chain reaction and DNA analyses, and only the unaffected or carrier embryos were transferred to uterus. Pregnancy confirmation, and prenatal diagnosis were done at 20 week's gestation.

RESULTSA total of 13 embryos were analyzed in the IVF cycle. PGD indicated that 2 were normal 18.1 , 3 were affected 27.3 , and 6 were carriers 54.5 ; diagnosis was not possible in 2. Three embryos were transferred to uterus on the third day after oocyte retrieval. Ultrasonography showed twin pregnancy with one blighted ovum. The prenatal diagnoses revealed that both fetuses were unaffected, one normal baby and one carrier were born.

CONCLUSIONThese studies represent the successful application of PGD for beta-thalassemia in China.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Male ; Mutation ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis ; methods ; Prenatal Diagnosis ; methods ; beta-Thalassemia ; diagnosis ; genetics ; prevention & control

OBJECTIVETo study the difference and similarity between Hans and Uighurs in regard to Rhesus box and its significance.

METHODSThe sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed on the basis of RHD gene sequence. The upstream, downstream and hybrid Rhesus boxes were determined by polymerase chain reaction-sequence specific primer(PCP-SSP) and mismatched PCR.

RESULTSThe percentage of RHD-/RHD-, RHD+/RHD- and RHD+/RHD+ genotypes ascertained in the unrelated Hans with RhD(-) were 61.40%, 34.21% and 4.39% respectively, while those in the unrelated Chinese Uighurs with RhD(-) were 94.44%, 2.78% and 2.78% respectively. Furthermore, all 6 cases of some other minorities were RHD-/RHD- types. The percentage of RHD-/RHD- and RHD+/RHD- genotypes ascertained in the unrelated Chinese Uighurs were significantly higher than those in Chinese Hans (P < 0.01), whereas no statistically significant difference in the percentage of RHD+/RDH+ genotype between the two groups was observed (P > 0.05).

CONCLUSIONThe Rh blood group of Uighurs in Xingjiang possesses both Oriental and Caucasian characteristics, which embodies a special ethnical aspect of the Chinese nation and is in accord with the anthropologic research results.

China ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics

OBJECTIVETo explore the expression and activation of NF-kappaB in middle ear cholesteatoma.

METHODSThe protein expression of NF-kappaB p65 in 21 middle ear cholesteatoma tissues and 8 normal external ear canal skin obtained in middle ear surgery were examined by immunohistochemistry; NF-kappaB DNA binding activity in these two kinds of tissues were also detected by electrophoretic motility shift assay (EMSA). The influence of cholesteatoma debris on the NF-kappaB DNA binding activity of HaCat cell were further analyzed.

RESULTSAll epithelial cell of cholesteatoma revealed a relatively abundant plasma expression of NF-kappaB p65 protein, among which 12 cases showed nuclear positive expression. In contrast,the normal skin epithelium only revealed a sparse plasma distribution of NF-kappaB protein. The levels of NF-kappaB p65 protein expression in the epithelium of middle ear cholesteatoma tissue and normal skin were 0.168 +/- 0.051, 0.088 +/- 0.019 (t = 4.211, P < 0.01), respectively. The NF-kappaB DNA binding activities of cholesteatoma [(16.5 +/- 10.1)%] were also higher than those in normal skin [(1.38 +/- 1.24)%, t = 3. 600, P = 0.014]. The NF-kappaB DNA binding activity of HaCat cell increased when exposed to cholesteatoma debris in a dose dependent manner.

CONCLUSIONNF-kappaB might be an important factor which was involved in the occurrence and development of cholesteatoma.

Adolescent ; Adult ; Biopsy ; Case-Control Studies ; Child ; Cholesteatoma, Middle Ear ; metabolism ; pathology ; Electrophoretic Mobility Shift Assay ; Female ; Humans ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Young Adult

BACKGROUNDClinical programs for preventing beta-thalassemia are presently based on prospective carrier screening and prenatal diagnosis. This paper report an achievement of a pregnancy with unaffected embryos using in vitro fertilization and embryo transfer (IVF-ET), in combination with preimplantation genetic diagnosis (PGD), for a couple at risk of having children with beta-thalassemia.

METHODSA couple carrying different thalassemia mutations, both a codon 41 - 42 mutation and the IVS II 654 mutation, received standard IVF treatment, with intracytoplasmic sperm injection, embryo biopsiy, single cell polymerase chain reaction (PCR) and DNA analysis. Only unaffected or carrier embryos were transferred to the uterine cavity. After confirmation of pregnancy, a prenatal diagnosis was performed.

RESULTSOf a total of 13 embryos analyzed for beta-globin mutations, PGD indicated that 2 were normal, 3 were affected, and 6 were carriers. Diagnosis could not be made in the other 2 embryos. Three embryos were transferred to the uterus on the third day after oocyte retrieval. Ultrasonography revealed a twin pregnancy with one blighted ovum. The prenatal genetic diagnosis revealed that both fetuses were unaffected, and two healthy boys were born, confirming the results of PGD.

CONCLUSIONSWe developed a single-cell based primer extension preamplification (PEP)-PCR assay for the detection of beta-thalassemia mutations. The assays were efficient and accurate at all stages of the procedure, and resulted in the birth of PGD-confirmed beta-thalassemia free children in China. PEP was used here in PGD for beta-thalassemia.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infant, Newborn ; Male ; Mutation ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis ; beta-Thalassemia ; diagnosis ; genetics