Comparative study on wound healing and infection between open and minimally invasive surgical methods in pediatric otolaryngology surgery.

Pediatric otolaryngology surgeries are crucial interventions requiring careful consideration of surgical methods to optimize outcomes. The choice between open and minimally invasive surgical approaches in this context warrants thorough investigation. While both methods aim to address ear, nose, and throat conditions in children, a comparative study assessing their impact on crucial factors such as intraoperative parameters, wound healing, complications, and postoperative pain is essential. This study aims to compare the effects of open and minimally invasive surgical methods on wound healing and infection in pediatric otolaryngology surgery, and provide a scientific basis for the selection of surgical methods. Two groups of patients were selected, with 90 people in each group. One group received open surgery and the other received minimally invasive surgery. Recording the intraoperative time, anesthesia time, and intraoperative blood loss; the number of days required for wound healing; the occurrence of wound-related complications; the comparison of pain on postoperative Days 1, 3, and 7; and the factors influencing postoperative wound healing were analyzed. In the minimally invasive surgery group, the intraoperative time was shorter, the anesthesia time was relatively reduced, and the amount of bleeding was significantly reduced. Wounds also take fewer days to heal and have lower rates of wound-related complications. When comparing the pain on 1, 3, and 7 days after surgery, the minimally invasive surgery group had relatively mild pain. Analysis of postoperative wound healing factors showed that minimally invasive surgical methods have a positive impact on healing. In pediatric otolaryngology surgery, minimally invasive surgery performs better than open surgery in terms of intraoperative operation time, anesthesia time, blood loss, wound healing time, complication rate, and postoperative pain. Therefore, minimally invasive surgery may be a safer and more effective surgical method.

Preliminary investigation of the diagnosis and gene function of deep learning PTPN11 gene mutation syndrome deafness.

Syndromic deafness caused by PTPN11 gene mutation has gradually come into the public's view. In the past, many people did not understand its application mechanism and role and only focused on non-syndromic deafness, so the research on syndromic deafness is not in-depth and there is a large degree of lack of research in this area. In order to let the public know more about the diagnosis and gene function of deafness caused by PTPN11 gene mutation syndrome, this paper used deep learning technology to study the diagnosis and gene function of deafness caused by syndrome with the concept of intelligent medical treatment, and finally drew a feasible conclusion. This paper provided a theoretical and practical basis for the diagnosis of deafness caused by PTPN11 gene mutation syndrome and the study of gene function. This paper made a retrospective analysis of the clinical data of 85 deaf children who visited Hunan Children's Hospital,P.R. China from January 2020 to December 2021. The conclusion were as follows: Children aged 1-6 years old had multiple syndrome deafness, while children under 1 year old and children aged 6-12 years old had relatively low probability of complex deafness; girls were not easy to have comprehensive deafness, but there was no specific basis to prove that the occurrence of comprehensive deafness was necessarily related to gender; the hearing loss of patients with Noonan Syndrome was mainly characterized by moderate and severe damage and abnormal inner ear and auditory nerve; most of the mutation genes in children were located in Exon1 and Exon3, with a total probability of 57.65%. In the course of the experiment, it was found that deep learning was effective in the diagnosis of deafness with PTPN11 gene mutation syndrome. This technology could be applied to medical diagnosis to facilitate the diagnosis and treatment of more patients with deafness with syndrome. Intelligent medical treatment was also becoming a hot topic nowadays. By using this concept to analyze and study the pathological characteristics of deafness caused by PTPN11 gene mutation syndrome, it not only promoted patients to find diseases in time, but also helped doctors to diagnose and treat such diseases, which was of great significance to patients and doctors. The study of PTPN11 gene mutation syndrome deafness was also of great significance in genetics. The analysis of its genes not only enriched the gene pool, but also provided reference for future research.

Relationship between single nucleotide polymorphism of NOS2 gene and inheritance of allergic rhinitis in children.

Allergic rhinitis is a common chronic disease, and its high incidence has a great negative impact on the quality of life of many people, especially children. In this paper, through in-depth analysis of NOS2 gene polymorphism, the protective mechanism of NOS2 gene against AR was studied to provide theoretical and scientific basis for the diagnosis of children with AR. It was concluded that the concentration of Immunoglobulin E (IgE) in rs2297516 was 0.24 IU/mL compared with that in normal children. rs3794766 specific IgE concentration in the children group was increased by 0.36 IU/mL, which was higher than that in the healthy children group; the difference of rs7406657 specific IgE concentration between the children group and the healthy group was 0.03 IU/mL. The total serum IgE concentration in the healthy children group was lower than that in the infant group, and the change of Rs3794766 was the least, followed by rs2297516 and rs7406657. This means that rs7406657 is the highest, rs2297516 had general genetic correlation with AR patients, and rs3794766 had the least genetic correlation with AR patients. Among the three groups of SNP loci, the healthy children group was higher than the patient children group, indicating that AR reduces the gene frequency of the three loci, and the reduction of gene frequency will also increase the susceptibility of children to AR, because the frequency of gene occurrence will affect the gene sequence. In conclusion, smart medicine and gene SNPS can promote the detection and treatment of AR.

Long non-coding RNA MALAT1 promotes Th2 differentiation by regulating microRNA-135b-5p/GATA-3 axis in children with allergic rhinitis.

Allergic rhinitis (AR) threatens patient survival. CD4+ T cells play key roles in AR progression. Long non-coding RNAs (lncRNAs) are key regulators of cell differentiation. Therefore, we investigated the molecular mechanism of the lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in AR. Expression levels of MALAT1, microRNA (miR)-135b-5p, interleukin-4 (IL-4), and GATA-binding protein 3 (GATA-3) in the nasal mucosa of AR patients were quantified. CD4+ T cells were isolated from the peripheral blood of healthy volunteers and treated with ovalbumin (OVA) and Th2 inducers. After MALAT1 and miR-135b-5p levels changed in CD4+ T cells, the proportion of IL-4-expressing cells and the levels of IL-4 and GATA-3 in OVA-induced CD4+ T cells were determined. Binding relationships among MALAT1, miR-135b-5p, and GATA-3 were predicted and verified. Rescue experiments were performed to confirm the role of the MALAT1/miR-135b-5p/GATA-3 axis in Th2 differentiation of CD4+ T cells. MALAT1, IL-4, and GATA-3 expression was upregulated, whereas miR-135b-5p expression was downregulated, in patients with AR. MALAT1 knockdown or miR-135b-5p overexpression in CD4+ T cells notably decreased the proportion of IL-4-expressing cells and downregulated GATA-3 and IL-4 expression in OVA-induced CD4+ T cells. MALAT1 and GATA-3 exhibited competitive binding toward miR-135b-5p. MALAT1 facilitated CD4+ T cell Th2 differentiation via the miR-135b-5p/GATA-3 axis. MALAT1 facilitated AR development by facilitating CD4+ T cell Th2 differentiation via the miR-135b-5p/GATA-3 axis. This study may provide guidance for clinical treatment of AR.

Phosphatidylethanolamine-binding protein 1 (PEBP1) mediates the regulatory role of microRNAs (miRNAs)-205-5p in degranulation and histamine release.

miR-205-5p plays a vital role in the inflammation of allergic rhinitis (AR). The study is designed to investigate the effects and mechanism of miR-205-5p in AR in vivo and in vitro. An OVA-induced mice model and anti-DNP IgE-induced RBL-2H3 cell model were established. The pathological alterations in the nasal mucosa were evaluated by hematoxylin-eosin (HE) staining. IgE and histamine levels were detected by corresponding kits and the expressions of PEBP1, High mobility group box-1 (HMGB1) and Toll-like receptor 4 (TLR4) were detected by western blot. The association of miR-205-5p and PEBP1 was determined by dual-luciferase reported assay. ß-hexosaminidase activity was to evaluate the degranulation of RBL-2H3 cell. The pathological injury of nasal mucosa was significantly improved by miR-205-5p inhibition compared to AR mice. Following the treatment of miR-205-5p inhibitor, the levels of helper T cell (Th1) cytokines, interleukin (IL)-2 and interferon-γ (IFN-γ) were increased, while the levels of Th2 cytokines, IL-4 and IL-13, as well as the levels of IgE and histamine were markedly decreased in AR mice. We further found that miR-205-5P inhibition induced increased expression of PEBP1 and decreased expressions of HMGB1and TLR4. In vitro, miR-205-5P was verified to bind to PEBP1. PEBP1 silencing led to the reverse of miR-205-5p effects on decreasing the levels of ß-hexosaminidase activity and histamine, as well as the expressions of HMGB1 and TLR4 on anti-DNP IgE-induced RBL-2H3 cells. Our results indicate that miR-205-5P inhibition may ameliorate pathological injury via PEBP1. MiR-205-5P/ PEBP1 could be potential drug targets in AR.

Aberrant expressions of circulating lncRNA NEAT1 and microRNA-125a are linked with Th2 cells and symptom severity in pediatric allergic rhinitis.

OBJECTIVE: Long noncoding RNA nuclear enriched abundant transcript 1 (lnc-NEAT1) and its target microRNA-125a (miR-125a) are reported to regulate immune and inflammation process in allergic rhinitis (AR). Hence, this study intended to investigate the correlation between lnc-NEAT1 and miR-125a expressions, as well as their clinical values in pediatric AR patients. METHODS: Peripheral blood mononuclear cell samples from 80 pediatric AR patients, 40 disease controls (DCs), and 40 healthy controls (HCs) were collected to detect lnc-NEAT1 and miR-125a expressions by reverse transcription-quantitative polymerase chain reaction. For pediatric AR patients only, serum interferon-gamma (IFN-γ) and interleukin (IL)-10 were measured by enzyme linked immunosorbent assay; meanwhile, T helper (Th) 1 and Th2 cells in CD4+ T cells were analyzed by flow cytometry. RESULTS: Lnc-NEAT1 was overexpressed, while miR-125a downregulated in pediatric AR patients compared to DCs and HCs (all p < 0.001). Moreover, lnc-NEAT1 expression negatively correlated with miR-125a expression in pediatric AR patients (p = 0.002), but not in DCs (p = 0.226) or HCs (p = 0.237). Furthermore, in pediatric AR patients, lnc-NEAT1 expression positively associated with TNSS (p < 0.001), sneezing score (p = 0.006), and congestion score (p = 0.008); miR-125a expression was negatively related to TNSS (p < 0.001), itching score (p = 0.040), and sneezing score (p = 0.005). Additionally, lnc-NEAT1 expression positively, while miR-125a expression negatively correlated with Th2 cells and IL-10 (all p < 0.05), but they were not correlated with Th1 cells or IFN-γ in pediatric AR patients. CONCLUSION: Circulating lnc-NEAT1 and miR-125a are aberrantly expressed and linked with Th2 cells and symptom severity in pediatric allergic rhinitis.

Role of hypermethylated-lncRNAs in the prognosis of bladder cancer patients.

OBJECTIVE: To explore the hypermethylated long non-coding (lnc)RNAs involved in bladder carcinogenesis and prognosis. METHODS: Reduced representation bisulfite sequencing and RNA sequencing were performed on five paired tumor and adjacent normal tissue samples from bladder cancer patients. The differentially methylated regions around transcription start sites and differentially expressed genes, including lncRNAs, were analyzed. Correlations between DNA methylation modifications and the expression of lncRNAs were examined. Survival analysis was surveyed on the GEPIA web server. RESULTS: We identified 19,560 hypomethylated and 68,781 hypermethylated differentially methylated regions around transcription start sites in bladder cancer tissues. In total, 2321 differentially expressed genes were found in bladder tumors, among which, 367 were upregulated and 1954 were downregulated. There were 141 downregulated genes involving eight lncRNAs that were consistently hypermethylated, while 24 upregulated genes were consistently hypomethylated. Survival analysis demonstrated that hypermethylation of lncRNAs LINC00683 and MSC-AS1 were associated with poor overall survival in bladder cancer patients. CONCLUSION: Some lncRNAs are controlled by DNA methylation in bladder cancer and they might be important factors in bladder carcinogenesis. Hypermethylated lncRNAs including LINC00683 and MSC-AS1 have the potential to be prognostic biomarkers for bladder cancer.

Amphiphilic Double-Brush Copolymers with a Polyurethane Backbone: A Bespoke Macromolecular Emulsifier for Ionic Liquid-in-Oil Emulsion.

The study on ionic liquid (IL)-based emulsions is very interesting due to the "green" quality and potential wide applications of ILs, whereas the emulsifiers for the formation of IL-based emulsions are extremely limited and mainly centered on low molecular weight surfactants. In this work, synthesis of amphiphilic double-brush copolymers (DBCs) and their application as bespoke macromolecular emulsifiers for the formation of IL-containing non-aqueous emulsions are described. DBCs consisted of a polyurethane (PU) backbone and poly(N,N-dimethyl acrylamide) (PDMA) and poly(methyl methacrylate) (PMMA) chains that were grafted simultaneously at the same reactive site along the PU backbone (PU-g-PDMA/PMMA), which were synthesized through the combination of polyaddition and the reversible-deactivation radical polymerization reactions. Highly stable [Bmim][PF6]-in-benzene emulsions could be gained by adopting PU-g-PDMA/PMMA DBCs as macromolecular emulsifiers at a low content, such as 0.025 wt %. On the basis of the stability and the size of emulsion droplets, PU-g-PDMA/PMMA DBCs exhibited much better emulsifying performances than their analogues, including PU-g-PDMA, PU-g-PMMA, and PDMA-b-PMMA copolymers. Such excellent emulsifying performances of PU-g-PDMA/PMMA DBCs were due to high interfacial activities. PU-g-PDMA/PMMA DBCs exhibited higher capabilities in lowering the interfacial tension of the [Bmim][PF6]-benzene interface than their analogues. A large energy barrier to desorption of adsorbed PU-g-PDMA/PMMA DBCs from the interface contributed to high stability of the [Bmim][PF6]-in-benzene emulsion.

Janus-Like Single-Chain Polymer Nanoparticles as Two-in-One Emulsifiers for Aqueous and Nonaqueous Pickering Emulsions.

The exploration of Pickering emulsions is very significant owing to their versatile and important applications in many scopes. In this study, synthesis of a novel kind of single-chain polymer nanoparticle (SCPN) and its stabilized Pickering emulsions were demonstrated. To this end, linear-dendritic diblock copolymers consisting of poly((2-dimethylamino) ethyl methacrylate) (PDMAEMA) blocks and four-generation dendritic aliphatic polyester blocks (G4) have been first synthesized by the combination of click chemistry and reversible addition-fragmentation chain transfer (RAFT) polymerization reaction. The subsequent intramolecular cross-linking of the PDMAEMA block of PDMAEMA-b-G4 copolymers in DMF using 1,4-diiodobutane as cross-linkers afforded Janus-like SCPNs that exhibited a cross-linked PDMAEMA head tethered by a short dendritic tail. The molecular weight and distribution together with the structure of polymers were carefully characterized by GPC and NMR spectroscopy. By the employment of the as-synthesized Janus-like SCPNs as Pickering emulsifiers, aqueous and nonaqueous Pickering emulsions including water-in-oil and oil-in-oil as well as ionic liquid-in-oil were generated. Under the same conditions, it was found that the long-term stabilities of Pickering emulsions stabilized by Janus-like SCPNs were superior to those of Pickering emulsions stabilized by their linear quaternized PDMAEMA-b-G4 by CH3I analogous.

Giant posterior pararenal schwannoma: A case report and review of literature.

Schwannomas, arising from Schwann cells of peripheral nerve sheaths, are primarily benign tumors. They are rarely found in the retroperitoneal space. To date, ~30 cases on giant retroperitoneal schwannomas have been reported. Those with a location of pararenal space are even rarer. Clinically, they are often misdiagnosed as malignancies. In the present study, a case of a 35-year-old woman with a giant schwannoma measuring 13 × 8.5 × 6.5 cm in the posterior pararenal space of left retroperitoneum was presented, which was thought to be a malignant tumor from the result of computed tomography scan. Later postoperative pathology and immunohistochemistry were consistent with benign schwannoma. To the best of our knowledge, this was the largest posterior pararenal schwannoma reported in literature. It is significant to report this case, and summarize the characteristics, diagnosis, treatment and prognosis of the similar cases. Furthermore, a brief review of the previously published cases in literature is provided.

Oncogenic miR-425-5p is associated with cellular migration, proliferation and apoptosis in renal cell carcinoma.

An increasing number of studies have demonstrated the function of microRNAs (miRNAs) in the initiation and development of various types of cancer. Among them, miR-425-5p is proven to serve an important function in several types of cancer, including gastric, cervical cancer, and hepatocellular carcinoma. However, the function of miR-425-5p in renal cell carcinoma (RCC) remains unclear. In the present study, it was demonstrated that the expression level of miR-425-5p was upregulated in RCC tissues and cell lines compared with normal tissues and cell lines (P<0.05). Additionally, Cell Counting kit-8 and MTT assays were employed to assess cell viability and proliferation, whereas wound healing and Transwell assays were employed to examine migration and invasion. The results demonstrated that upregulation of miR-425-5p promoted cell viability and the invasion and migration of ACHN and 786O cells (P<0.05). Flow cytometric analysis confirmed that upregulation of miR-425-5p inhibited apoptosis of ACHN and 786O cells (P<0.05). Downregulation of miR-425-5p inhibited the viability and invasion and migration of ACHN and 786O cells (P<0.05). In the present study, upregulation of miR-425-5p inhibited apoptosis of ACHN and 786O cells whereas no differences in early apoptotic rate were observed between the inhibitor and inhibitor NC groups for 786O and ACHN cells. These results indicate that miR-425-5p may act as an oncogene in RCC.

Tuberous sclerosis-associated renal angiomyolipoma: A report of two cases and review of the literature.

In the general population, tuberous sclerosis-associated renal angiomyolipoma (RAML) is a rare benign hamartoma with potentially life-threatening complications and a poor prognosis. The two patients reported in the present study, who were diagnosed with bilateral RAML and spontaneous rupture of the RAML with tuberous sclerosis, presented with flank pain, abdominal pain, hemorrhage, hematuria and multiple lesions. The two cases are representative examples of the disease, and highlight the importance of determining the risk of acute hemorrhage in the early stages, and the significance of timely and proper treatment.

Oncogenic microRNA-142-3p is associated with cellular migration, proliferation and apoptosis in renal cell carcinoma.

MicroRNAs (miRNAs/miRs) serve an important role in the regulation of carcinogenic pathways. RCC is the most prevalent kidney cancer that occurs in adults. miRNAs have gained increasing attention due to their association with RCC tumorigenesis, serving as biomarkers for early detection and progression monitoring, and as potential targets for molecular therapy. Upregulation of miRNA-142-3p has been previously identified in RCC tissues by microarray profile, however, its expression and function in RCC have not yet been validated. In the present study, quantitative polymerase chain reaction was performed to quantify the relative expression of miR-142-3p in 53 paired RCC and adjacent normal tissues. Furthermore, wound healing, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry assays were performed to analyze the impacts of miR-142-3p on cellular migration, proliferation and apoptosis. The results demonstrated that miR-142-3p was significantly upregulated in RCC tissues compared with adjacent normal tissues. Downregulation of miR-142-3p, induced by chemically synthesized miR-142-3p inhibitor, significantly suppressed cell migration and proliferation, and promoted cell apoptosis in 786-O and ACHN cells, supporting the theory that miR-142-3p may function as an oncogene in RCC. The potential clinical significance of miR-142-3p, as a biomarker and therapeutic target, provides rationale for further investigation into the miR-142-3p-mediated molecular pathway and how it is associated with RCC development.

MicroRNA-106b functions as an oncogene in renal cell carcinoma by affecting cell proliferation, migration and apoptosis.

Kidney cancer is the 14th most common cancer in the world and its prognosis remains poor due to difficult early detection and treatment. Therefore, the identification of biomarkers for early-stage renal cell carcinoma (RCC) is important. MicroRNA-106b (miR-106b) has been described as an oncogene in several types of human cancer. Previous microarray studies have suggested that miR-106b was significantly upregulated in RCC tissues compared with paired normal kidney tissues and may be a promising biomarker for the prediction of early metastasis following nephrectomy. The present study aimed to determine the expression and function of miR-106b in RCC. The expression of miR-106b in RCC tissues and cells, and in paired normal tissues and cells was determined by reverse transcription quantitative polymerase chain reaction, based on the previous sequencing results of miRNAs. Furthermore, a wound scratch assay, MTT assay and flow cytometry were performed to examine the functions of miR-106b on cell migration, proliferation and apoptosis. The results demonstrated that miR-106b was upregulated in RCC tissues and cell lines compared with control normal tissues and cell lines. Downregulation of miR-106b with a synthesized inhibitor suppressed cell migration and proliferation and induced renal cancer cell apoptosis, suggesting that miR-106b can be characterized as an oncogene in RCC. To the best of our knowledge, the present study was the first to reveal that miR-106b is upregulated and affects cellular migration, proliferation and apoptosis in RCC. Further studies are required to examine the role and target genes of miR-106b in RCC.

Primary mesenchymal chondrosarcoma with bilateral kidney invasion and calcification in renal pelvis: A case report and review of the literature.

Mesenchymal chondrosarcoma (MC) is a rare malignant cartilaginous forming tumor. MC of the kidney is extremely rare, with only seven cases reported in the literature. The present study described the case of a 17-year-old male, who presented with sudden severe pain in the right flank and a high fever. Imaging studies demonstrated a large soft heterogeneous mass (7.8×9.5×15 cm) located between the liver and right kidney with no clear demarcation, and a well-demarcated mass (1.3×2.4 cm) with patchy dense calcification occupying the left renal pelvis. Following the diagnosis of a Wilms' tumor, the patient underwent a right radical nephrectomy and the pathological diagnosis was MC of the kidney. To the best of our knowledge, the current study presents the first case of MC with bilateral kidney invasion and calcification in the renal pelvis. In addition, the clinical, radiological and pathological features, and the management of this unusual neoplasm were discussed.

Theoretical studies on vicinal-tetrazine compounds: furoxano-1,2,3,4-tetrazine-1,3,5-trioxide (FTTO-α) and furoxano-1,2,3,4-tetrazine-1,3,7-trioxide (FTTO-ß).

The derivatives of 1,2,3,4-tetrazine may be promising candidates of high-energy density compounds and are receiving more and more attention. In this study, two 1,2,3,4-tetrazines, furoxano-1,2,3,4-tetrazine-1,3,5-trioxide (FTTO-α) and furoxano-1,2,3,4-tetrazine-1,3,7-trioxide (FTTO-ß), were theoretically studied. The geometrical structures in gas phase were studied at the B3LYP/6-311++G(d,p) level of density functional theory (DFT). The gas phase enthalpies of formation were calculated by the homodesmotic reaction method. The enthalpies of sublimation and solid phase enthalpies of formation were predicted with corrections of electrostatic potential method at the B3PW91/6-31G(d,p) level. The detonation properties were estimated with the Kamlet-Jacobs equations based on the predicted densities and enthalpies of formation in solid state. The available free space in the lattice was calculated to evaluate their stability. Calculations of potential energy surface and structure interconversion thermodynamics under different temperatures were carried out to further confirm their stability. FTTOs have better performance than HMX and FTDO but are easy to decompose to 5,6-dinitroso-v-tetrazine 1,3-dioxide. A synthesis route for FTTO-ß was proposed to provide a consideration for the further study. We believe FTTOs could be the key compounds to synthesize other v-tetrazines such as TTTO.

Identification of miR-7 as an oncogene in renal cell carcinoma.

MicroRNA-7 (miR-7) has been described as a tumor suppressor in several human cancers, but the results of a study to identify miRNAs associated with metastatic capability in breast cancer suggested that miR-7 may be characterized as an oncogene. The present study was to determine the expression and function of miR-7 in renal cell carcinoma. Quantitative real-time polymerase chain reaction was used to validate the expressions of miR-7 in 48 paired renal cell carcinomas (RCC) and normal tissues, based on the preliminary sequencing results of miRNAs. Furthermore, the impacts of miR-7 on cell migration, proliferation and apoptosis were analyzed using wound scratch assay, MTT and flow cytometry, respectively. The results demonstrated that miR-7 was up-regulated in RCC compared with normal tissues (p = 0.001). Down-regulation of miR-7 with synthesized inhibitor inhibited cell migration in vitro, suppressed cell proliferation and induced renal cancer cell apoptosis, prompting that miR-7 could be characterized as an oncogene in RCC. The present study was the first to reveal that miR-7 was up-regulated in RCC and it played an important role in RCC by affecting cellular migration, proliferation and apoptosis. Further researches should be conducted to explore the roles and target genes of miR-7 in RCC and other cancers.

[Construction of SOD1 eukaryotic expression vector and its expression].

AIM: to construction of eukaryotic expression vector of the human SOD1 (Cu/Zn superoxide dismutase, SOD1) and expression in HeLa cells. METHODS: the open reading frame (ORF) of SOD1 was amplified from human peripheral blood by RT-PCR. TA cloning strategy was used to insert the target fragments into pUCm-T vector. The recombinant plasmid was identified and noted as pUCm-T-SOD1. Then, SOD1 was subcloned into pTracer-CMV/Bsd, a eukaryotic expression vector. The plasmid of pTracer-CMV/Bsd-SOD1 was sequenced and was introduced into HeLa cells by Lipofectamine(TM); 2000. The expression of green fluorescence protein (GFP) was observed by the fluorescence microscope. The expression of SOD1 was detected by RT-PCR and Western blot after screening by blasticidin for 4 weeks. RESULTS: the eukaryotic expression plasmid pTracer-CMV/Bsd-SOD1 was successfully constructed. The GFP was observed in transfected cells by the fluorescence microscope. The expression of SOD1 was detected in transfected cells by RT-PCR and Western blot. CONCLUSION: the recombinant eukaryotic expression vector of pTracer-CMV/Bsd-SOD1 has been constructed successfully which could express GFP and SOD1, respectively, providing a tool for further gene therapy study.

[Surface modification and DNA-binding assessment of nano-hydroxyapatite].

OBJECTIVE: To evaluate the impact of surface modification on the DNA-binding ability of nano-hydroxyapatite (nHA). METHODS: Chemical co-precipitation-hydrothermal synthesis was utilized to prepare the nHA particles, and polyethylenimine (PEI) was used for surface modification of the nHA. Transmission electron microscopic (TEM) observation and zeta potential detection of the nHA were carried out before and after surface modification. The abilities of the nanoparticles, at different pH values and different concentrations, for DNA-binding and DNA protection against nuclease digestion were assessed before and after surface modification by electrophoresis. RESULTS: TEM observation showed a short rod-like morphology of PEI-modified nHA with uniform particle size and good dispersion; the nHA without the modification tended to aggregate with poor dispersion. With a positive zeta potential, the PEI-modified nHA showed an obviously enhanced ability of DNA binding at different pH values and concentrations, with strong capacity to protect the DNA against Dnase I digestion. At the concentration of 250 µg/ml and a pH value of 7.0, the nHA-PEI showed an optimal efficiency of DNA-binding and DNA protection. CONCLUSION: nHA with surface modification by PEI can serve as an effective vector for DNA binding and transfer.