Objective To explore the effects of applying microsurgical technique in the one-stage urethroplasty for hypospadias with longitudinal preputial island flaps of dorsum penis. Methods Forty-two hypospadias cases with obvious phallocampsis were reviewed. The patients aged from 1 to 19 years (median, 6.4 years). Among them, 6 cases were of coronary suleus type, 28 cases of pe-nis body type, and 8 cases of penis and scrotum type. All patients went through the operative proce-dures by microscope with four-times magnification and microsurgical instruments. Longitudinal prepu-tial island flaps of dorsum penis was employed for the treatment of 33 cases of these patients, and 9 cases underwent longitudinal preputial island flaps of dorsum penis combined scrotal septal pedicle flaps. Results Thirty-eight cases were cured by one-time surgery (90.5%). One case with urethral fistula after operation was cured by surgical repair of urethral fistula. Two cases with urethral meatus stricture after operation and one with urethral stoma stricture were cured by dilatation. Patients were followed for 9 to 52 months (mean, 27 months). Conclusion Use of microsurgical technique with longitudinal preputial island flaps of dorsum penis in one-stage urethroplasty for hypospadias improved the success rate of surgery with few complications.

Oxidative stress is a redox imbalance in the body, which is one of the important factors leading to tissue damage and diseases. The nuclear factor E2-related factor 2 (Nrf2)-Kelch like ECH-associated protein 1 (Keap1) signaling pathway is not only an important defense system against oxidative damage, but also one of the key signaling pathways of the antioxidant capacity. Numerous studies have shown that targeting the Keap1-Nrf2 signaling pathway to activate Nrf2 has become an effective strategy for the treatment of oxidative stress and related diseases. Using small molecules to directly block the Keap1-Nrf2 protein-protein interaction (PPI) is one of the important directions for activating Nrf2 and exerting the cytoprotective effect, which can avoid the potential side effects of covalent modification of Nrf2. On the other hand, the Keap1 is an efficient E3 ubiquitin ligase that has been used in the design of proteolysis targeting chimeras (PROTACs). This review summarizes the research progresses of Keap1-Nrf2 protein interaction inhibitors and degraders based on the Keap1 E3 ubiquitination system in recent years.

Rotundine (1 micromol L(-1)) was incubated with a panel of rCYP enzymes (1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remained parent drug in incubates was quantitatively analyzed by an Agilent LC-MS. CYP2C19, 3A4 and 2D6 were identified to be the isoenzymes involved in the metabolism of rotundine. The individual contributions of CYP2C19, 3A4 and 2D6 to the rotundine metabolism were assessed using the method of total normalized rate to be 31.46%, 60.37% and 8.17%, respectively. The metabolites of rotundine in incubates were screened with ESI-MS at selected ion mode, and were further identified using MS2 spectra and precise molecular mass obtained from an Agilent LC/Q-TOF-MSMS, as well as MS(n) spectra of LC-iTrap-MS(n). The predominant metabolic pathway of rotundine in rCYP incubates was O-demethylation. A total 5 metabolites were identified including 4 isomerides of mono demethylated rotundine and one di-demethylated metabolite. The results also showed that CYP2C19, 2D6 and 3A4 mediated O-demethylation of methoxyl groups at different positions of rotundine. Furthermore, the ESI-MS cleavage patterns of rotundine and its metabolites were explored by using LC/Q-TOF-MSMS and LC/iTrap-MS(n) techniques.
Analgesics, Non-Narcotic ; metabolism ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Berberine Alkaloids ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2C19 ; Cytochrome P-450 CYP2C9 ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dopamine Antagonists ; metabolism ; Humans ; Isoenzymes ; metabolism ; Methylation ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Electrospray Ionization

Glucose-6-phosphate dehydrogenase deficiency (G6PD) is one of hereditary diseases sariously influencing the human health. G6PD is characterized by wide distribution, high incidence, inducing the hemolysis, complex mechanism of hemolysis and common occurence in children and so on. The blood transfusion is most effective method for acute ouset of hemolysis, but the risk is more high, thereby it is necessary to guarante the safety of blood transfusion. In addition of the routine managementin blood transfusion and standard procedures, the basic rules and indicators of blood transfusion must be grasped, the blood preparations should be known well, the blood protection must be strengthened, and the measures of personalized psychologic interference must be drawn up carnestly and performed strictily so as to improve the therapeutic efficacy and safe of blood transfusion. Through the promotion and development of social pubbicity, elucation activities, preventive and control measures and medical levels, many patients have been freed from healthy problems as soon as possible. In this review, the research progress on acute hemolysis and safe blood transfusion in G6PD are summarized.

[Objective]To discuss the CT diagnosis and differential diagnosis of solid pseudopapillary tumor of the pancreas (SPTP).[Methods]The CT findings of 20 patients with SPTP proved by surgically pathology were retrospectively analyzed and summa?rized.[Results]SPTP were composed of solid and cystic components with surrounding capsule resulting to clear demarcation between tumor and normal pancreas without dilation of pancreatic duct. The tumor parenchyma was slightly hyperenhancement on arterial phase and showed gradual enhancement on venous and delayed phase.[Conclusions]The CT findings of SPTP have relative specifici?ty and can contribute to early diagnosis and differential diagnosis of SPTP.

OBJECTIVE: To explore the causes and specific conditions of blood donation reaction under the collective emergency unpaid blood donation, and to provide theoretical basis and decision-making reference for drafting the collective emergency unpaid blood donation and blood donation safety. METHODS: Through a combination of prospective and retrospective models, and statistical methods were used to analyze the causes and conditions of the blood donation response of 10401 people participating in collective emergency unpaid blood donation during 2016.1-2018.8. RESULTS: A total of 10401 person-times donated blood in a sitting manner, and a total of 293 blood donation reactions occurred. By improving the blood donation services year by year, the moderate blood donation reaction during the year 2017 and 2018 was significantly lower than that in 2016 (P＜0.05). In the actual blood donation group of≤100, 200, 300 and 400 ml, the incidence of blood donation reaction was statistically significant (P＜0.05); the incidence of blood donation reaction in the blood donors for 1,2,3 and ＞3 drnations was also statistically significant (P＜0.05); the blood donation reactions rate of B antigen containers was significantly different from the donors without B antigen (P＜0.05); the incidence of blood donation reaction with related to the weight of the donor. CONCLUSION The blood donation reaction of collective emergency unpaid blood donation closely relates with mental factors, blood donation service, blood donation frequency and body weight of the blood donor. The first blood donation is more likely to produce blood donation reaction. The blood donation volum≤ 100 ml from blood donors is resulted mostly from blood donation reactions.
Blood Donors ; Blood Group Antigens ; Humans ; Prospective Studies ; Retrospective Studies

OBJECTIVETo observe the effect of crystalline NiS on genome DNA methylation profile in in vitro cultured cells.

METHODS16HBE Cells were treated with crystalline NiS at 0.25, 0.50, 1.00 and 2.00 µg/cm(2) for 24 h and three times at total. DAC treatment was given at 3 µmol/L for 72 h.5-mC immunofluorescence and SssI methyltransferase assay methods were applied to investigate if the hypomethylation of genome DNA involved.

RESULTSThe results of 5-mC immunofluorescence showed that the fluorescence intensity of NiS-treated cells were decreased in some degree, and transformed cells were decreased dramatically. By the SssI methylase assay, an average of (81.9 ± 7.3)% methylated CpG were found in negative control cells. By contrast, (77.9 ± 6.2)%, (75.3 ± 6.8)%, (59.5 ± 4.9)%, (67.4 ± 5.1)% methylated CpG were observed in cells treated with NiS for three times at dosage of 0.25, 0.50, 1.00 and 2.00 µg/cm(2) which were abbreviated as NiS0.25, NiS0.50, NiS1.00, NiS2.00 respectively. The ANOVA analysis results showed that there was a significant difference in the 5 groups above (F = 124.95, P < 0.01). The results of Dunnett-t test showed that the methylated CpG of both group NiS1.00 and NiS2.00 were significantly decreased compared with the negative control group (t values were 7.64, 4.89 respectively, P < 0.01). For methylated CpG, (46.2 ± 4.1)% and (43.6% ± 4.3)% were observed in NiS-transformed cells (NSTC1 and NSTC2) which were dramatically decreased compared with the negative control group (t values were 12.79, 13.56 respectively, P < 0.01).

CONCLUSIONGenomic DNA methylation levels were decreased during NiS induced malignant transformation.

Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; DNA Methylation ; drug effects ; Epithelial Cells ; drug effects ; Genome ; Humans ; Nickel ; adverse effects ; chemistry

BACKGROUNDSurvivin is a rather specific gene in tumor tissue. We transfected dendritic cells (DCs) with recombinant adenovirus (Ad) containing survivin gene and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene and tested the inducing effect of the transfected DCs on cytotoxic T lymphocytes (CTL) to kill leukemic cells.

METHODSAfter derived from the peripheral, DCs was assayed by mixed leukocyte reaction (MLR) tests. Lactate dehydrogenase (LDH) release test was used to evaluate cytotoxicity of CTL.

RESULTSExpression of survivin in transfected DCs was confirmed by Western blotting analysis. GM-CSF expression was confirmed by enzyme-linked immunosorbent assay (ELISA). In MLR assay, DCs coinfected with Ad-survivin and Ad-GM-CSF induced higher allogeneic lymphocyte reaction than control DCs at ratios of 1:5, 1:10, 1:50 and 1:100. DCs coinfected with Ad-survivin and Ad-GM-CSF had much higher activity of CTL to HL-60 cells than DCs infected with Ad-survivin only, Ad-GM-CSF only, or control DCs. Levels of interleukin-12 (IL-12) and interferon gamma (IFN-gamma) in lymphocyte supernatants containing DCs coinfected with Ad-survivin and Ad-GM-CSF were significantly higher than those in the control group.

CONCLUSIONDCs coinfected with Ad-survivin and Ad-GM-CSF induce much higher anti-leukemic response in vitro than those infected with either factor. Therefore, adenovirus vectors containing survivin and GM-CSF genes may be promising vaccine candidates for leukemia therapy.

Adenoviridae ; genetics ; Cytotoxicity, Immunologic ; Dendritic Cells ; physiology ; ultrastructure ; Genetic Therapy ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; HL-60 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; Interferon-gamma ; biosynthesis ; Interleukin-12 ; biosynthesis ; Leukemia ; therapy ; Lymphocyte Activation ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection

OBJECTIVETo detect gene mutations associated with autosomal dominant congenital stationary night blindness(ADCSNB) in a large Chinese family.

METHODSGenomic DNAs were extracted from peripheral blood samples of 16 affected and 14 unaffected family members. According to 5 missense mutations in 3 genes reported previously, 4 pairs of primers were designed and corresponding exons containing the five mutation sites were amplified by polymerase chain reaction. Amplified products were purified and sequenced by MegaBACE1000 capillary array electrophoresis DNA sequencer. Full field electroretinogram (ERG, ISCEV) of patients was recorded and analyzed by Roland Consult System.

RESULTSDark-adapted ERG showed a-wave was normal, but b-wave of the patients was markedly decreased. None of the five missense mutations were detected in 16 affected and 14 unaffected family members.

CONCLUSIONThe molecular pathogenesis of ADCSNB in this family does not involve point mutations or deletions of these five sites, which indicates the heterogeneity of ADCSNB.

Adult ; Base Sequence ; DNA Mutational Analysis ; Female ; Humans ; Male ; Molecular Sequence Data ; Night Blindness ; congenital ; genetics ; Pedigree ; Point Mutation

OBJECTIVETo explore the effects of bisphenol A (BPA) exposure on toxicity characteristic and OCT4 and SOX2 gene expression of mouse embryonic stem cells (mESC).

METHODSmESC were cultured, and treated with the doses of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L respectively of BPA and DMSO (the solvent control group)for 24 hours, and three groups of cells were treated with the same method. The morphological changes of mESC in the control and exposure groups were observed through an inverted microscope. Cell counting kit 8 (CCK8) was used to detect the effects of BPA on proliferation of mESC, and based on the results, the half inhibitory concentration (IC50) was calculated. Real-time fluorescent quantitative polymerase chain reaction (RT-QPCR) and western blotting were used to detect the expression of OCT4 and SOX2.

RESULTSBPA had certain toxicity on mESC, the treatment of BPA significantly increased cell toxicity in a concentration-dependent manner, and the IC50 was 4.3×10(-4) mol/L, combined with the BPA exposure concentration of the environment and the related literature, eventually taking the five concentrations of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L as the experimental groups. The mESC morphology were effected after the treatment of BPA for 24 h, compared with the control group, the number of cells decreased, appearing some floating cells, and the cell cloning became irregular and differentiation in the higher concentration groups. The OCT4 mRNA expression level in the 10(-7) mol/L (1.146 ± 0.087), 10(-6) mol/L (1.156 ± 0.030), 10(-5) mol/L (1.158 ± 0.103) and the 10(-4) mol/L (1.374 ± 0.053) dose group were all significantly higher than the control group (1.000 ± 0.000) (t values were -2.384, -2.953, -3.203, -4.021 respectively, P value all < 0.05). Meanwhile, the SOX2 mRNA expression level in the 10(-4) mol/L (1.113 ± 0.052) were higher than the control group (1.000 ± 0.000) (t value was -2.765, P value < 0.05). Moreover, the OCT4 protein expression level in the 10(-5) mol/L (1.360 ± 0.168) and 10(-4) mol/L (1.602 ± 0.151) were all significantly higher than the control group (1.000 ± 0.000) (t values were -3.538, -4.002 respectively, P value all < 0.05), while no obvious change of the SOX2 protein expression level was detected in all treated groups.

CONCLUSIONBPA in a certain dose range could upregulate the expression of OCT4 gene in mouse embryonic stem cells while had no significant effect on the expression of SOX2 gene.

Animals ; Benzhydryl Compounds ; toxicity ; Cells, Cultured ; Embryonic Stem Cells ; drug effects ; metabolism ; Gene Expression ; Mice ; Octamer Transcription Factor-3 ; genetics ; Phenols ; toxicity ; SOXB1 Transcription Factors ; genetics ; Signal Transduction ; drug effects