To examine the significance of heat shock protein 70 mRNA in ototoxicity resulted from gentamicin (GM), twenty healthy albino guinea pigs (200-250 g) of either sex with a positive Prier reflex were divided into two groups randomly. In GM group the animals received 100 mg/kg GM daily by intraperitoneal injection for 10 d. In saline control group the animals received 2.5 ml/kg saline daily by intraperitoneal injection for 10 d. Auditory brainstem response (ABR) thresholds were recorded in each animal before and 1 d after GM or saline administration. After the second ABR measurement, the expression of HSP70 mRNA in guinea pig cochlea was observed with in situ hybridization and image quantitative analysis system. The results showed that the threshold of ABR in the GM group was significantly higher than that of the saline control (P< 0.001). The expression of HSP70 mRNA was more intensive in stria vascularis, spiral ligament and spiral ganglion cells in the GM group than that of the saline control group. These results suggest that administration of gentamicin can induce the expression of HSP 70 mRNA in guinea pig cochlea, and that this effect may protect hearing function from ototoxicity.
Animals ; Cochlea ; metabolism ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Gentamicins ; toxicity ; Guinea Pigs ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation

AIMTo explore the effects of gentamicin ototoxicity on the expression of heat shock protein 70 in guinea pigs cochlea.

METHODSWe used immunohistochemistry staining and image quantitative analysis system, combined with auditory brainstem response (ABR) measurement to investigate the change on the expression of HSP70 in guinea pigs cochlea of gentamicin ototoxicity.

RESULTSThe levels of HSP70 immunoreactivity in guinea pigs cochlea of experimental animals were high including Corti's organ, stria vascularis, medial spiral limbus, spiral ganglion cells and the threshold of ABR was in high correlation with the expression of HSP70 ([ r] > 0.8, P < 0.01).

CONCLUSIONGentamicin can induce expression of HSP 70 in guinea pigs cochlea and protect hearing function.

Animals ; Cochlea ; drug effects ; physiopathology ; Gentamicins ; toxicity ; Guinea Pigs ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; drug effects

OBJECTIVETo globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogenesis system with Affymetrix oligonucleotide array.

METHODSA microcarrier-based in vitro angiogenesis system was developed, in which endothelial cells (ECs) migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed network. The total RNA samples from the HMVECs at the selected time points (0.5 h, 24 h, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the software provided by the manufactory. The expression patterns of some genes were validated and confirmed by Semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines/chemokine receptors were specially examined and their functional implications were analyzed.

RESULTSAbout 1500 genes were found up- or down- regulated 2-folds or above detected by the arrays, and among them, about 400 genes regulated 3-folds or above. The regulated genes could be grouped into categories based on their molecular functions such as growth factor and receptor, cell proliferation, extracellular matrix, cell cycle and apoptosis, signaling molecule and transcription factor, and so on, using the Gene Ontology Mining Tool in The NetAffx Analysis Center. The regulated genes were also clustered into six groups based on their patterns of expression. As for chemokines, the CCL2/MCP-1, CCL5/RANTES and CX3CL1 were identified to be specially upregulated at 24 h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis.

CONCLUSIONSBased on our angiogenesis model, and by oligonucleotide arrays, the present study demonstrates global profiles of the gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be encouraged.

Capillaries ; cytology ; Cells, Cultured ; Chemokines ; genetics ; Endothelial Cells ; cytology ; Endothelium, Vascular ; cytology ; physiology ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; In Vitro Techniques ; Neovascularization, Physiologic ; genetics ; Oligonucleotide Array Sequence Analysis ; Receptors, Chemokine ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the correlation of the CYP1A1 (rs4646422) gene polymorphisms with male infertility in the Chinese Han population.

METHODSUsing the Mass ARRAY iPLEX GOLD technique, we conducted a case-control study on theCYPlA1 (rs4646422) gene polymorphisms in 636 infertile males aged 21-49 years (case group) and 442 normal healthy men aged 23-47 years (control group) of the Chinese Han population. We analyzed the genotypes and allele frequencies in the two groups ofsubjects with the SPSS 20.0 software.

RESULTSCompared with the wild homozygous genotype GG, the heterozygous genotype AG (OR = 1.06, 95% CI 0.81-1.38) and homozygous genotype AA (OR = 1.11, 95% CI 0.56-2.21) showed no correlation with male infertility, nor did the mutant allele A (OR = 1.06, 95% CI 0.85-1.32) in comparison with the wild allele G.

CONCLUSIONThe CYP1A1 (rs4646422) gene polymorphisms might not be correlated with male infertility in the Chinese Han population.

Adult ; Alleles ; Case-Control Studies ; China ; Cytochrome P-450 CYP1A1 ; genetics ; Gene Frequency ; Genotype ; Homozygote ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).

METHODSFrom January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.

RESULTSOf 48 patients, 15 were transplanted from unrelated donors with an antigen mismatch for HLA Cw and 33 patient-donor pairs were matched for HLA-Cw. The CMV reaction rate was 66.7% for HLA-Cw mismatch group and 48.5% for HLA-Cw match group (chi(2) = 1.39, P = 0.2375). Thirty-seven donor-patients pairs belonged to group C1 and 11 to group C2, and CMV reaction rate was 64.9% in group C1 and 18.2% in group C2 (chi(2) = 18.13, P < 0.0001). Twenty-six patients received graft from KIR haplotype A (group A donor) and 22 from KIR haplotype B donors (group B donor) and CMV reaction rate was 57.7% in group A donor and 50.0% in group B donor (chi(2) = 0.28, P = 0.5941). The number of donor activating KIRs (aKIRs) was less than that of recipient aKIRs in 34 patient-donor pairs in which the CMV reaction rate was 70.6%, and the number of donor aKIRs was more than that of recipient aKIRs in 14 patient-donor pairs in which the CMV reactivation was 14.3%. There was a significan difference between the two group (chi(2) = 12.44, P = 0.0004).

CONCLUSIONKIR and HLA-Cw genotypes influence the rate of CMV reactivation following non-T cell deleted unrelated donor hematopoietic cell transplantation.

Genotype ; HLA-C Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics

ObjectiveTo evaluate the clinical efficacy of sequential syndrome differentiation of Yiqi Huayu Qingre prescription （YHQ） in the treatment of refractory nephrotic syndrome in children. MethodA total of 112 children with refractory nephrotic syndrome were randomly divided into an observation group （57 cases） and a control group（55 cases）. The children in the control group were treated with prednisone tablets combined with tacrolimus，and those in the observation group were treated with YHQ by sequential syndrome differentiation on the basis of the control group. The total effective rates of the two groups after treatment were observed. The 24-hour urinary total protein（24 h UTP），plasma albumin（ALB），cholesterol（CHO），triglycerides（TG）， and traditional Chinese medicine quality of life scale scores before treatment and after four weeks，eight weeks，16 weeks，24 weeks，32 weeks，40 weeks，and 52 weeks in the two groups were recorded. The total course of treatment and the total accumulation of hormones were compared among the children with reduced or no hormone treatment till 52 weeks during treatment. ResultThe total effective rate in the observation group was higher （Z=-2.052，P<0.05）. The observation group had lower 24 h UTP and higher ALB at each follow-up time point than the control group（P<0.05，P<0.01）. At four weeks，eight weeks，and 16 weeks of treatment，there was no statistically significant difference in CHO between the observation group and the control group，and the observation group was lower than the control group in CHO at the rest of the time points （P<0.05，P<0.01）. For TG， the observation group was not significantly different from the control group at four weeks，eight weeks，16 weeks，and 40 weeks of treatment，but lower at 24，32，and 52 weeks （P<0.05，P<0.01）. The total treatment course of hormones in the observation group was shorter（P<0.01）， with less total accumulation（P<0.01）. At different follow-up time points，the total score of traditional Chinese medicine quality of life scale in the observation group was superior to that in the control group（P<0.05，P<0.01），and the scores of the observation group in the four dimensions （physiological function，independent factor，social factor，and psychological factor） after treatment were higher than those in the control group（P<0.05，P<0.01）. ConclusionYHQ under sequential syndrome differentiation has a definite clinical effect in treating children with refractory nephrotic syndrome. It has advantages in shortening the total course of hormone treatment and reducing the total accumulation of hormones，and can improve the quality of life of children with refractory nephrotic syndrome.

This study was purposed to investigate the clinical value of HLA matching(low and high resolution) and its effect on outcome of the patients received umbilical cord blood transplantation(UCBT). Sequence-specific oligonucleotide probe (SSOP) , sequence-based typing (SBT) and sequence-specific primers(SSP) were used to perform high resolution HLA matching for HLA-A, -B, -Cw, -DRB1, -DQB1 and low resolution for HLA-A, B, DRB1 among 34 patients with hematologic malignancies who received unrelated UCB transplantation and grafts. The effects of HLA matching (low or high resolution ) on leading engraftment, hematopoietic reconstitution, graft-versus-host disease (GVHD) and infection after UCB transplantation were analyzed by comparison. The results showed that the median of total nucleated cells (TNC) of transplanted cord blood was 6.0×10(7)/kg, The time of neutrophil recovery was significantly shortened when more than 5×10(7)/kg TNC were transplanted (P < 0.05). The HLA-(6-10)/10 group of high resolution HLA matching was better than the HLA (4-5)/10 group in the respect of leading engraftment, the time of platelet recovery and the rate of acute GVHD (P < 0.05). In contrast, HLA-I+II locus, HLA-DRB1 or HLA-DQB1 locus mismatch could prolong the platelet engraftment time (P < 0.05). There was statistical difference in the time of platelet recovery, the rate of acute GVHD between the HLA (5-6)/6 group of low resolution HLA matching and the HLA (3-4)/6 group after UCB transplantation (P < 0.05), but the mismatch locus of HLA with low resolution did not correlate with the time of platelet recovery (P > 0.05). It is concluded that the high resolution HLA matching between patients received unrelated UCB transplantation and grafts may contribute to select the better UCB, that has important clinical value to promote hematopoietic reconstitution and to reduce the complications after UCB transplantation.
Adolescent ; Adult ; Child ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Hematologic Neoplasms ; therapy ; Histocompatibility Testing ; methods ; Humans ; Male ; Young Adult

Epigenetic abnormalities play an important role in the pathogenesis of hematological malignancies, especially acute leukemia (AL). Similar to DNA methylation and histone modifications, RNA methylation is another important epigenetic modification. m6A methylation is one of the most prevalent and extensively studied RNA methylation. m6A methylation is involved in many biological and pathological process. Recent studies have found that m6A methylation is involved in the occurrence, development and drug-resistance of AL. This review focuses on the research progress of m6A methylation in AL.
DNA Methylation ; Epigenesis, Genetic ; Humans ; Leukemia

Resistance to cisplatin (DDP)-based chemotherapy is a major cause of treatment failure in human gastric cancer (GC). It is necessary to identify the drugs to re-sensitize GC cells to DDP. In our previous research, Zuo Jin Wan Formula (ZJW) has been proved could increase the mitochondrial apoptosis via cofilin-1 in a immortalized cell line, SGC-7901/DDP. Due to the immortalized cells may still difficult highly recapitulate the important molecular events in vivo, primary GC cells model derived from clinical patient was constructed in the present study to further evaluate the effect of ZJW and the underlying molecular mechanism. Immunofluorescent staining was used to indentify primary cultured human GC cells. Western blotting was carried out to detect the protein expression. Cell Counting Kit-8 (CCK-8) was used to evaluate cell proliferation. Flow cytometry analysis was performed to assess cell apoptosis. ZJW inhibited proliferation and induced apoptosis in primary DDP-resistant GC cells. Notably, the apoptosis in GC cells was mediated by inducing cofilin-1 mitochondrial translocation, down-regulating Bcl-2 and up-regulating Bax expression. Surprisingly, the level of p-AKT protein was higher in DDP-resistant GC cells than that of the DDP-sensitive GC cells, and the activation of AKT could attenuate ZJW-induced sensitivity to DDP. These data revealed that ZJW can increase the chemosensitivity in DDP-resistant primary GC cells by inducing mitochondrial apoptosis and AKT inactivation. The combining chemotherapy with ZJW may be an effective therapeutic strategy for GC chemoresistance patients.
Adult ; Aged ; Aged, 80 and over ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; therapeutic use ; Cofilin 1 ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Mitochondria ; drug effects ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured

Objective: To investigate the correlation of the single nucleotide polymorphism (SNP) rs1042522 of the tumor protein p53 (TP53) gene with the risk of male infertility. METHODS: This casecontrol study included 380 male patients with idiopathic infertility and 398 normal fertile men as controls from the Nanjing area. We genotyped the SNP rs1042522 of the TP53 gene by Sequence Mass Array and analyzed the correlation of the SNP with male infertility using the logistic regression model. RESULTS: Compared with the normal controls, the patients with idiopathic infertility showed significantly decreased sperm concentration (［77.34±49.24］ vs ［13.13±24.96］ ×106/ml), percentage of progressively motile sperm (［42.55±9.57］ vs ［10.38±5.57］%), serum testosterone level (［14.07±5.36］ vs ［11.89±4.50］ nmol/L), and folliclestimulating hormone level (［16.80±18.20］ vs ［4.55±7.17］ U/L) (P < 0.05) but no statistically significant differences in other parameters. No correlation was observed between the SNP frequencies and male infertility and similar results were found in the subgroups of the cases. CONCLUSIONS SNP rs1042522 of the TP53 gene is not significantly correlated with the risk of male infertility.
Case-Control Studies ; Follicle Stimulating Hormone ; blood ; Gene Frequency ; Genes, p53 ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infertility, Male ; blood ; genetics ; Logistic Models ; Male ; Polymorphism, Single Nucleotide ; Sperm Count ; Sperm Motility ; Testosterone ; analogs & derivatives ; blood