Reconstruction of large defects of anterior floor of mouth with free flaps using a novel individualized flap design method.

The purpose of this study was to introduce a novel individualized flap design method for large anterior floor of the mouth (AFOM) defect reconstruction, review experience with the use of this flap design method for large AFOM defect reconstruction, and assess its functional results. A retrospective study of patients who received large AFOM defect reconstruction with free flaps was conducted. There was a cohort of patients who were treated using the novel individualized flap design method and a cohort without flap design. Functional outcomes were evaluated with appropriate scales. Outcomes were analyzed, and a p-value <0.05 was considered significant. 22 patients received the individualized flap design, while 21 patients were treated without a special flap design. All flaps survived. All free flaps harvested with the novel individualized flap design method better matched AFOM defects. Relative to patients without flap design, patients in the novel individualized flap design group showed significant improvement in speech intelligibility (p = 0.036) and swallowing function (p = 0.019). Within the limitation of the study it seems that large AFOM defect reconstruction with the novel individualized flap design method can not only cover and close the wound to avoid oral-neck fistulae, but also maintains tongue mobility to achieve better functional outcomes than in patients without flap design.

A novel flap design technique for subtotal tongue reconstruction with an "Individualized and Convenient Tongue Model".

BACKGROUND: To achieve improved functional outcomes in subtotal tongue reconstruction, a flap design with sufficient volume and appropriate shape is necessary. In this study, we introduce an "Individualized and Convenient Tongue Model" (ICTM) for flap design in subtotal tongue reconstruction. METHODS: By studying the anatomical morphology of the tongue, we found a similar geometry within the dorsum and body of the tongue as well as the mouth floor. This can be used to create an ICTM through folding and splicing. We can simulate tongue defects in the ICTM and transform defect shapes into guide plates for flap design. In this study, fifty-eight patients requiring subtotal tongue reconstruction were randomly divided into two groups: an ICTM group (35 patients) and a conventional group (31 patients). In the ICTM group, we individually designed profunda artery perforator flaps (PAPFs) or anterolateral thigh flaps (ALTFs) using the ICTM method. In the conventional group, the flap was designed according to the surgeon's clinical experience. Patient demographics, operative and follow-up data were recorded. Swallowing, speech intelligibility, and cosmetic results were assessed using appropriate scales. RESULTS: All flaps survived, although there were no significant differences in tumor size, operation time, flap size, and complication rate compared to the conventional group. Patients in the ICTM group had significantly improved speech intelligibility (p = 0.019), cosmetic appearance (p = 0.009), and swallowing ability (p = 0.003). CONCLUSIONS: The ICTM technique is an effective and convenient solution for subtotal tongue reconstruction that provides an individualized flap design and improves functional outcomes compared to the conventional design.

[Construction of Vectors Expressing Inhibiting Peptides for Nuclear Import and Its Effect on Growth and Migration of HeLa Cell].

OBJECTIVE: To construct the expression vectors for red fluorescent protein fused with inhibiting peptides for nuclear import (Bimax),and explore the location of Bimax and its potential effects on cell proliferation and migration in HeLa cells. METHODS: Two kinds of polynucleotide encoding inhibiting peptides for nuclear import were synthesis respectively and subsequently annealed for inserting into vector pDs-Red-C1. The recombinant plasmids were transfected into competent bacterial DH-5α. After transfection,the positive bacteria were picked up for DNA sequencing. The recombinant plasmids pDs-Red-Bimax2,pDs-Red-Bimax1 and negative plasmid pDs-Red-C1 were transfected into HeLa cells respectively according to Lipofectamine2000 protocol. After transfection,the expression and location of red fluorescent protein were observed with fluorescence microscope. Furthermore,MTT assay and cell-migration assay were used to detect the proliferation and migration of Bimax transducted cells. RESULTS: DNA sequencing showed that the polynucleotides encoding Bimax1 or Bimax2 were inserted into pDs-Red-C1 vector successfully. After transfected into HeLa cells,the inhibiting peptide induced red fluorescent protein locating in nuclear. Furthermore,either the fusion protein RFP-Bimax1 or RFP-Bimax2 can suppress the proliferation and migration of HeLa cells. CONCLUSION: The expression vectors for red fluorescent protein fused with inhibiting peptides for nuclear import were successfully constructed. In addition,the fusion proteins were expressed and located in nuclear and suppressed the proliferation and migration of tumor cells.

[Emissions of greenhouse gas and ammonia from the full process of sewage sludge composting and land application of compost].

There is a great uncertainty of greenhouse gas (GHG) reduction and nitrogen conservation from the full process of sludge composting and land application of compost in China due to the lack of emission data of GHG such as N2O and CH4 and ammonia (NH3). The purpose of this study is to get emission characteristics of GHGs and NH3 from the full process with on-site observation. Results showed that the total GHG emission factor from full process of the turning windrow (TW) system (eCO2/dry sludge, 196.21 kg x t(-1)) was 1.61 times higher of that from the ATP system. Among the full process, N2O was mostly from the land application of compost, whereas CH4 mainly resulted from the sludge composting. In the sludge composting of ATP, the GHG emission equivalence of the ATP (eCO2/dry sludge, 12.47 kg x t(-1) was much lower than that of the TW (eCO2/dry sludge, 86.84 kg x t(-1)). The total NH3 emission factor of the TW (NH3/dry sludge, 6.86 kg x t(-1)) was slightly higher than that of the ATP (NH3/dry sludge, 6.63 kg x t(-1)). NH3 was the major contributor of nitrogen loss in the full process. During the composting, the nitrogen loss as NH3 from both TW and ATP was nearly the same as 30% of TN loss from raw materials, and the N and C loss caused by N2O and CH4 were negligible. These results clearly showed that the ATP was a kind of environmentally friendly composting technology.

[Construction and identification of eukaryotic expression plasmid expressing siRNA targeting USP22 gene].

OBJECTIVE: To construct eukaryotic expression plasmid expressing siRNA targeting ubiquitin specific peptidase 22 gene (USP22), and to investigate its effect on the growth of hepatoma carcinoma cells HepG2. METHODS: siRNA templates were synthesized based on USP22 mRNA sequence and cloned into vector Pmscv/Hyg/U6. The resulting recombinant was identified by restriction enzyme digestion and DNA sequencing. Recombinants were than transfected into HepG2 cells mediated by liposome. The USP22 protein and mRNA in HepG2 cells were detected by western blot and RT-PCR, respectively. The cellular growth activity was evaluated with MTT assay. RESULTS: Recombinant plasmid expressing siRNA targeting USP22 was successfully constructed. The down-regulated protein and mRNA level of USP22 and decreased cellular growth in HepG2 cells transfected with recombinant plasmid were observed. CONCLUSION: The eukaryotic expression vector for RNA interference USP22 gene is constructed successfully, which inhibits the expression of USP22 in HepG2 cells and suppresses cell proliferation.

[Kinetic characteristics of Cr(III) oxidation by delta-MnO2].

The kinetics of Cr(III) oxidation by synthesized vernadite(delta-MnO2) was investigated through magnetic stirring and fractional centrifugation. The oxidation procedure of Cr(III) by delta-MnO2 could be divided into two first-order reactions, a fast reaction followed by a slow one. In high Cr(III) concentration solution, the reaction was also well sub-simulated with diffusion equation and Elovich equation according to the reaction phases. The rate constants markedly increased with temperature rising. The ratio of Cr(VI)/Mn(II) gradually decreased and eventually reached the theoretical value (0.667) with the reaction. Cr(VI), Cr(III) and Mn(II) adsorbed on delta-MnO2 only accounted 0.1 to 3% of the total amounts. The Cr(VI)/Mn(II) ratio on the surface of MnO2 was much smaller than the theoretical value due to the release of Mn(II) to the solution lagged behind Cr(VI). When Cr(III) was in low concentration, Cr(III) oxidation was controlled by the diffusion and adsorption of Cr(III). While in high Cr(III) concentration, the key step was the diffusion of Mn(II) to solution.

Silencing of Ref-1 expression by retrovirus-mediated shRNA sensitizes HEK293 cells to hydrogen peroxide-induced apoptosis.

Redox factor-1 (Ref-1) is a multifunctional protein involved in DNA base excision repair (BER) and transcription factor modification. By the use of retrovirus-delivered shRNA, we effectively suppressed endogenous Ref-1 expression in human embryonic kidney (HEK) 293 cells. Our results showed that downregulation of Ref-1 rendered cells more sensitive to H(2)O(2)-induced apoptosis. In this process, the absence of Ref-1 decreased the ratio of Bcl-2/Bax protein expression, which resulted in cytochrome c release and caspase-3 activation. These data indicate that constitutive Ref-1 is required for cellular defense and that this protective function is dependent on its role in the regulation of Bcl-2 family proteins.

A soluble truncated cadherin induces breast cancer cell apoptosis and growth inhibition.

PURPOSE: To identify the characteristics and function of the truncated cadherin cDNA which encodes a soluble molecule containing the sequence of VE-cadherin extracellular domain repeats from repeat 1 to 4 (designated as CED1-4) and a secreting signal peptide at N terminal. METHODS: A pMSCV/CED1-4 vector was constructed. Recombinant retrovirus ReCED1-4 and ReEmpty were produced by 293 package cells and transfected into MDA-MB435 human breast cancer cells. The expression of CED1-4 in transfectants and their supernatant was analyzed by RT-PCR and Western blot, respectively. MDA-MB435 cell proliferation assays were performed in vitro and in vivo. CED-14-induced apoptosis was demonstrated using Annexin V binding, TUNEL and caspase 3 assays. The expression of integrin beta1 and c-fos mRNA was detected by RT-PCR. RESULTS: The constructed soluble CED1-4 encoded 484 amino acids and a secreting signal peptide (27 amino acids). CED1-4 was expressed by MDA-MB435/CED1-4 cells, and detected in the supernatant of CED1-4 tranfectants. CED1-4 transfection significantly inhibited the growth of MDA-MB435 cells in vitro and in vivo. About 22-fold increase in the early apoptotic cells in MDA-MB435/CED1-4 cells was observed as compared with MDA-MB435/empty cells. Increased activity of caspase 3 in MDA-MB435/CED1-4 cells was more than two times as compared with that of the control cells. Interestingly, integrin beta1 transcriptional level in MDA-MB435/CED1-4 cells was down-regulated as compared with control cells. The resistance of fibronectin to CED1-4 apoptotic inducibility was confirmed by detection of caspase 3. The blockage of c-fos transcriptional expression was detected in MDA-MB435/CED1-4 cells. CONCLUSIONS: The soluble truncated cadherin may be considered an apoptotic inducer and growth inhibitor in the MDA-MB435 breast carcinoma cell line. Down-regulation of integrin beta1 and blockage of c-fos expression may be related to CED1-4-induced apoptosis and growth inhibition.