Objective To explore the efficacy of transplantation of a U-shaped ilioinguinal flap in the re pair of skin and soft tissue defects of the extremity.An axial flap based on the superficial iliac circumflex artery and trimmed to a subdermal vascular network flap was used for the procedures.Methods Seven patients with skin and soft tissue defects treated between June,2009 and May,2014 were studied.The patients were 22-45 years of age (mean,32 years),and included 5 males and 2 females.Four patients had punch-press injuries,1 patient had a hot-crush injury,and the remaining 2 patients were injured in the accidents.The wound sizes were 14.0 cm × 10.0 cm to 6.0 cm × 5.0 cm,with a varying extent of exposed tendons and bones.Repairs were performed using free ilioinguinal flaps,which were 15.0 cm × 11.0 cm to 7.0 cm × 5.0 cm in size.The axial flap was trimmed to a U-shaped subdermal vascular network flap and transplanted to the recipient site with anastomosis of blood vessels.Results All transplanted flaps survived.Four patients were followed for 1-6 months,with a mean duration of follow-up of 4 months.The trimmed flaps showed gradual reddening immediately after surgery,and the capillaries were recovered with a flat surface.Re-examination 3 months after surgery showed that the flaps were thin and flexible and met the aesthetic demand.No obvious pigmentation occurred,and the donor site was sutured directly,leaving only linear scars.Conclusion Repair of skin and soft tissue defects of the extremity using a U-shaped trimmed ilioinguinal flap has the advantages of a hidden donor site,small scar,and conformity to aesthetic requirements.The trimmed flaps are preferred over untrimmed flaps in terms of color and texture.The former flap is thinner,meets the aesthetic demand,and achieves a better efficacy.

Objective To investigate the effect and mechanism of terazosin combined with Qianlielongbitong in the treatment of non-bacterial prostatitis.Methods One hundred and two patients with non-bacterial prostatitis were divided into two groups:one group was treated with terazosin 4 mg qn and Qianlielongbitong,while the other group was treated with terazosin and Levoofloxacin.We compared three indices of chronic prostatitis symptom index (NIH-CPSI),prostatic secretion examination(EPS) and urodynamic data in three different steps:before treatment,after 4-week treatment and after 8-week treatment.Results The NIH-CPSI of both groups was greatly improved after treatment( all P ＜0.01 ).Inside the treatment group,the NIH-C-PSI after 8-week treatment was better than that after 4- week treatment ( all P ＜ 0.01 ).However,in both groups,there was no significant difference between the index after 8-week treatment and the one after 4 week.EPS,AFR and MFR were greatly improved in both groups( all P ＜0.01 ).Conclusion Terazosin can relieve the clinical symptom,and improve the life quality.

Objective To discuss the significance of the retrograde urethrography in diagnosis and treatment of urethraltrauma. Methods 78 cases with urethraltrauma treated by the retrograde urethrography were retrospectively analyzed. Results The location and extent of urethral injury was determined according to the place and speed of contrast medium overflow and the diffuse range. Among 78 cases ,29 cases were bulbar urethral trauma and other 49 cases were membranous urethral trauma.Conclusion Retrograde urethrography is simple, practical and easy to operate for determining the injured part of urethra and the extent of damage of urethraltrauma, and was instructional for the choice of operation method and incision.

Objective To evaluate the efficacy and safety of hydroxychloroquine combined with butyli flufenamatum ointment and other drugs for the treatment of polymorphous light eruption (PLE).Methods A total of 48 patients with PLE were randomly and equally classified into group 1 and group 2.Both groups took hydroxychloroquine 200 mg twice a day and loratadine 10 mg per day for the initial 4 weeks,then took hydroxychloroquine 100 mg twice a day alone for another 4 weeks.Group 1 also topically applied butyli flufenamatum ointment twice a day during the 8 weeks,while group 2 applied mometasone furoate cream twice a day for the first 2 weeks followed by butyli flufenamatum ointment twice a day for another 6 weeks.Each treatment cycle lasted 2 weeks,and both groups received 4 cycles of treatment.Patients were evaluated for the response rate at the end of each cycle,and for the total symptom score and erythema score before and after the 8-week treatment.Statistical analysis was carried out using t test,chi-square test,Fisher's exact test and repeated-measures analysis of variance with the SPSS17.0 software.Results On day 14,28,42 and 56,the total score improved in 0,3,12 and 19 patients in group 1 respectively,and in 1,4,12 and 20 patients in group 2 respectively;the erythema score improved in 1,5,13 and 18 patients in group 1 respectively,and in 0,5,11 and 17 patients in group 2 respectively.No significant difference was observed between the two groups in response rates at any of the above four time points (P ＞ 0.05).Both the total score and erythema score significantly decreased after the 8-week treatment in both groups compared with the pretreatment scores (both P ＜ 0.05).No serious adverse reaction was observed in either of the two groups.Conclusions Hydroxychloroquine combined with loratadine and butyli flufenamatum ointment shows high efficacy and safety for the treatment of PLE.Topical butyli flufenamatum ointment is highly effective for the treatment of PLE,especially for PLE cases mainly presenting with erythema.

OBJECTIVETo investigate retinoblastoma (Rb) associated protein 46 (RbAp46) gene expression levels in bone marrow (BM) cells of leukemia patients.

METHODSReal-time quantitative reverse polymerase chain reaction (QRT-PCR) method was used for detecting RbAp46 expression levels in BM cells of 140 patients with acute leukemia (AL), 13 with chronic myelogenous leukemia in chronic phase (CML-CP), 7 with CML in blast crisis (CML-BC) and 32 with non-leukemic disorders.

RESULTSThe M-Estimators of RbAp46 were higher in 98 newly diagnosed ALs and 5 relapsed ALs than in 28 ALs in complete remission (CR) and 32 non-leukemic controls (178.23 and 213.65 vs 85.89 and 88.08, respectively). No statistic difference was found between the CR group and control group, or between the newly diagnosed group and relapsed group. The M-Estimators of RbAp46 in patients with CML-CP was 58.27, similar to that in control, but much lower than that in CML-BC (173.24). Among 98 newly diagnosed ALs, the M-Estimators of RbAp46 in M(3) and M(4) were the lowest in all of the subtypes. Furthermore, the RbAp46 expression levels were not correlated with the expression of the fusion genes of bcr/abl, PML-RARalpha, and multidrug resistant gene (mdr1), but were positively correlated with Wilms' tumor gene (WT1) expression levels and negatively with AML1/ETO fusion gene expression.

CONCLUSIONRbAp46 expression levels in ALs and CML-BC were strikingly higher than that in non-leukemias and CML-CP, and might participate in leukemogenesis.

Adolescent ; Adult ; Aged ; Carrier Proteins ; genetics ; metabolism ; Child ; Female ; Gene Expression ; Humans ; Leukemia ; genetics ; metabolism ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; Retinoblastoma-Binding Protein 7 ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Acute Disease ; Blast Crisis ; genetics ; Genes, Retinoblastoma ; Genes, Wilms Tumor ; Humans ; Leukemia ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; pathology ; Polymerase Chain Reaction ; U937 Cells

To explore the feasibility of real-time quantitative PCR (QRT-PCR) for selecting cell strains which overexpress a certain transgene, expression level of RbAp46 was detected in transfected cell strains by using optimal real-time PCR with SYBR Green I. Meanwhile, semi-quantitative RT-PCR and Western blot were performed to compare with the QRT-PCR. The results showed that values of RbAp46(N) were 2064.42 +/- 253.47, 860.94 +/- 291.07, 234.456 +/- 31.08, 18.17 +/- 5.14 and 1.46 +/- 0.54 in K562/RbAp46, K562/CMV, SHG44/RbAp46 monoclone, SHG44/RbAp46 multiclone and SHG44/CMV, respectively. The results were consistent with that determined by semi-quantitative RT-PCR and Western blot. It is concluded that QRT-PCR provides a highly efficient and reproducible method for selection of transfected cell subclones at different level of transgene expression.
Blotting, Western ; Carrier Proteins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; Nuclear Proteins ; genetics ; metabolism ; Organic Chemicals ; chemistry ; RNA, Neoplasm ; metabolism ; Retinoblastoma-Binding Protein 7 ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transfection ; Transgenes ; genetics

OBJECTIVETo study the DNA methylation status of WT1 gene promoter region in hematologic malignancy cell lines and its correlation with WT1 gene expression.

METHODS1. RT-PCR and methylation-specific PCR were performed for detecting WT1 gene expression and DNA methylation status in its promoter region in 8226, HL-60, Jurkat, K562, KG-1, NB4, Raji, SHI-1, U266 and U937 cell lines. 2. Treatment of U937 cells with 5-aza-CdR, a demethylation inducing agent and the changes in WT1 gene expression level and its promoter region methylation status were determined.

RESULTS1. HL-60, K562, KG-1, NB4 and SHI-1 cells showed higher levels while 8226, Jurkat, Raji, U266 and U937 cells showed extremely low levels of WT1 expression. DNA hypermethylation in WT1 gene promoter region was identified in 8226, Jurkat, Raji, U266 and U937 cells. 2. The WT1 gene expression in U937 was enhanced after treatment with 5-aza-CdR accompanied with the decrease of methylated and the increase of unmethylated levels in its promoter region.

CONCLUSIONModulation of the DNA methylation status in WT1 promoter region is one of the epigenetic mechanisms for regulating its expression.

Cell Line, Tumor ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hematologic Neoplasms ; genetics ; Humans ; Promoter Regions, Genetic ; genetics ; WT1 Proteins ; genetics

The objective of study was to investigate tissue-specific transcriptional activity of WT1 (Wilms' tumor gene) promoter and enhancer in cell lines with diverse tissue origin for leukemic gene therapy depending on WT1 transcriptional regulation elements. WT1 promoter and enhancer were ligated into pEGFP-1 to construct a recombinant vectors with EGFP gene as a reporter. By using electroporation or lipofectamine, the resultant constructs were transfected into 13 cell lines including WT1-expressing hematopoietic cell lines (K562, NB4, THP-1 and SHI-1), WT1-nonexpressing hematopoietic cell lines (U937 and Jurkat), WT1-expressing nonhematopoietic cell lines (MCF-7, T47D and 293) and WT1-nonexpressing nonhematopoietic cell lines (ECV304, SMMC7721, HT-29 and SHG44). The mean fluorescence intensity (MFI) of EGFP representing the transcriptional activities of promoter and/or enhancer was analyzed by using flow cytometry in the transfected cells which stably expressed EGFP. The results indicated that the vectors, pEWP containing WT1 promoter and pEWPA containing both WT1 enhancer and promoter, were constructed by recombinant DNA technique. Among nonhematopoietic cell lines, pEWP induced the highest EGFP expression in ECV304 (16.54 +/- 2.45 times as high as pEGFP-1), mildly higher in MCF-7 and SHG44 (9.46 +/- 1.10 and 7.29 +/- 0.73 times of pEGFP-1 level), and lowest in HT-29 (0.99 +/- 0.02 times as much as pEGFP-1) respectively. Among hematopoietic cell lines, EGFP expression was highest in K562 cell line (2.93 +/- 0.27 times of pEGFP-1), which was statistically higher than those in Jurkat and SHI-1 cell lines (0.74 +/- 0.03 and 0.84 +/- 0.09 times of pEGFP-1 level) respectively. pEWPA, with WT1 enhancer inserted at Afl II site near SV40 polyA, increased basal transcription levels of the WT1 promoter in HT-29, SHI-1 and K562 cells by 4.81, 3.06 and 1.01-fold respectively. It is concluded that the transcriptional activities of WT1 promoter in the recombinant vector seem unrelated to the constitutional expression level of endogenous WT1 gene. The WT1 enhancer promotes the transcriptional activities of WT1 promoter in some of the cell lines regardless of the hematopoietic tissue origin.
Enhancer Elements, Genetic ; genetics ; Humans ; Jurkat Cells ; K562 Cells ; Promoter Regions, Genetic ; genetics ; Transcription, Genetic ; U937 Cells ; WT1 Proteins ; genetics ; metabolism

OBJECTIVETo investigate Wilms' tumor gene (WT1) expression levels in bone marrow (BM) of acute leukemia patients (ALs).

METHODSA real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method was established for detecting WT1 and internal reference GAPDH expression levels in BM of 108 ALs and 23 non-leukemia controls by Light Cycler.

RESULTSThe median expression levels of WT1 in 70 newly diagnosed ALs and 11 relapsed ALs were statistically higher than those in 23 ALs in complete remission (CR) and 23 non-leukemic controls (75.10 and 89.56 vs 2.07 and 1.51 respectively). No statistic differences was found between the CR group and control group, nor between the newly diagnosed group and relapsed group. Of the 70 newly diagnosed ALs, median WT1 expression level of acute granulocytic leukemias was significantly higher than that of acute monocytic leukemias (M(5)), but there was no statistic differences among the M(1), M(2), M(3) and ALL subtypes. Furthermore the WT1 levels were not correlated to peripheral WBC counts, BM blast percentage and multidrug resistant gene (mdr1) expression at presentation, but correlated to chromosome karyotypes. Dynamic analysis of WT1 levels of 2 patients on treatment showed that WT1 expression levels predicted relapse.

CONCLUSIONWT1 expression levels in ALs were strikingly higher than that in non-leukemias. WT1 can be a marker for detecting MRD and evaluating therapy efficacy in leukemias.

Acute Disease ; Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Child ; Female ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia ; blood ; genetics ; Leukemia, Monocytic, Acute ; blood ; genetics ; Leukemia, Myeloid ; blood ; genetics ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; WT1 Proteins ; genetics ; Young Adult