Locally advanced breast cancer patients should be cautious about the immediate breast reconstruction after mastectomy: a pooling analysis of safety and efficacy.

BACKGROUND: The purpose of this study was to compare safety and efficacy outcomes between immediate breast reconstruction (IBR) and mastectomy alone in locally advanced breast cancer patients. METHODS: We conducted a comprehensive literature search of PUBMED, EMBASE, and Cochrane databases. The primary outcomes evaluated were overall survival, disease-free survival, and local recurrence. The secondary outcome was the incidence of surgical complications. All data were analyzed using Review Manager 5.3. RESULTS: Sixteen studies, involving 15,364 participants were included in this meta-analysis. Pooled data demonstrated that patients underwent IBR were more likely to experience surgical complications than those underwent mastectomy alone (HR: 3.96, 95%CI [1.07,14.67], p = 0.04). No significant difference was found in overall survival (HR: 0.94, 95%CI [0.73,1.20], p = 0.62), disease-free survival (HR: 1.03, 95%CI [0.83,1.27], p = 0.81), or breast cancer specific survival (HR: 0.93, 95%CI [0.71,1.21], p = 0.57) between IBR group and Non-IBR group. CONCLUSIONS: Our study demonstrates that IBR after mastectomy does not affect the overall survival and disease-free survival of locally advanced breast cancer patients. However, IBR brings with it a nonnegligible higher risk of complications and needs to be fully evaluated and carefully decided.

Tailoring Efficient Fluorogenic Tactic for Ultrasensitive Detection of Dopamine in Urine and Rat Brain through Real-Time and In Situ Formation of High-Performance Fluorophore.

Owing to the predominance of dopamine (DA) in controlling mental health, planning an innovative method for DA detection with simplicity and high efficacy is conducive to the assessment of neurological disorders. Herein, an efficient fluorogenic tactic has been elaborated for ultrasensitive detection of DA with remarkably enhanced turn-on response. Utilizing a twisted intramolecular charge-transfer (TICT)-suppressing strategy, a highly emissive azocine derivative 11-hydroxy-2,3,6,7,11,12,13,14-octahydro-1H,5H,10H-11,14a-methanoazocino[5',4':4,5]furo[2,3-f]pyrido[3,2,1-ij]quinolin-10-one (J-Aza) is generated via a one-step reaction between DA and 8-hydroxyjulolidine. It is marvelous that J-Aza not only possesses ideal fluorescence quantum yield (ΦF) as high as 0.956 but also exhibits bathochromic shifted fluorescence (green emissive) and stronger anti-photobleaching capacity superior to traditional azocine-derived 1,2,3,4-tetrahydro-5H-4,11a-methanobenzofuro[2,3-d]azocin-5-one (Aza) with moderate ΦF, blue fluorescence, and poor photostability. By confining the TICT process, the detection limit to DA can be reduced to 80 pM, which is competitive in contrast to previously reported fluorescence methods. Encouraged by the instant response (within 90 s), wide linear range (0.1-500 nM), great selectivity, and excellent sensitivity, this fluorogenic method has been used for the real-time measurement of DA contents in practical urine samples with satisfactory results. Furthermore, the cerebral DA level in the reserpine-induced depression rat model has also been evaluated by our designed method, demonstrating its potent analytical applicability in the biosensing field.

Controllable atomic collision in a tight optical dipole trap.

Single atoms are interesting candidates for studying quantum optics and quantum information processing. Recently, trapping and manipulation of single atoms using tight optical dipole traps has generated considerable interest. Here we report an experimental investigation of the dynamics of atoms in a modified optical dipole trap with a backward propagating dipole trap beam, where a change in the two-atom collision rate by six times has been achieved. The theoretical model presented gives a prediction of high probabilities of few-atom loading rates under proper experimental conditions. This work provides an alternative approach to the control of the few-atom dynamics in a dipole trap and the study of the collective quantum optical effects of a few atoms.

Management Strategy for Gallbladder Polypoid Lesions: Results of a 5-Year Single-Center Cohort Study.

INTRODUCTION: Controversy remains about the classification, differential diagnosis, and treatment strategy for gallbladder polypoid lesions (GPLs). This study sought to explore the individualized treatment strategy for GPLs. METHODS: We retrospectively studied 642 consecutive patients with GPLs from January 2015 to May 2020. Univariate and multivariable analyses were performed to explore the potential risk factors for neoplastic polyps. The outcome of laparoscopic gallbladder-preserving polypectomy (GPP) was evaluated and compared with that of laparoscopic cholecystectomy (LC). RESULTS: Of 642 enrolled patients, 572 underwent LC, and 70 underwent GPP. Pathologically, the majority of GPLs were cholesterol polyps (68.4%), followed by adenomyomatosis (19.9%), benign adenoma (7.3%), adenocarcinoma (3.6%), and rare pathological types (0.8%). Additionally, 66.3% (379/572) of the LC cases were classified as non-neoplastic, and 33.7% (193/572) neoplastic polyps. Multivariate analysis demonstrated that single polyps (OR 1.956, 95% CI: 1.121-3.412; p = 0.018), polyps located at the gallbladder fundus (OR 4.326, 95% CI: 2.179-8.591; p < 0.001), polyps not less than 14 mm (OR 2.833, 95% CI: 1.614-4.973; p < 0.001), and polyps with a broad base (OR 4.173, 95% CI: 1.743-9.990; p = 0.001) were independent risk factors for neoplastic polyps. The 5-year prospective results after GPP showed that the 1-year and 3-year polyp recurrence rates were 13.2% and 23.4%, respectively. CONCLUSION: The majority of GPLs are cholesterol or other benign lesions without malignant potential. LC is the main treatment procedure for GPLs with a high neoplastic risk. GPP is potentially feasible and could be an alternative management strategy for a group of GPLs patients who meet the selection criteria.

Stabilizing a laser frequency by the Pound-Drever-Hall technique with an acousto-optic modulator.

We develop and demonstrate a method of optical phase modulation in the Pound-Drever-Hall (PDH) technique. The phase modulation in this paper is realized by an acousto-optic modulator (AOM) operating in the Bragg diffraction regime. In this process, a light beam separated from a laser (780 nm) is sent through the AOM twice and coupled to a high finesse Fabry-Perot cavity. Then, the light power coupling into the cavity is stabilized by modulating the optical amplitude with this AOM. The coupling light power is stabilized to a level of 10-3. In the meantime, the PDH error signal is obtained by modulating the optical phase with the same AOM. After the error signal is fed back to the laser current, the laser linewidth is suppressed to approximately 907.91 Hz. This method of phase modulation is simple and convenient, and we believe it can be widely used in modulation transfer spectroscopy and frequency-modulation sideband spectroscopy.

High-resolution ex vacuo objective for cold atom experiments.

We present a versatile and cost-efficient objective with a five-lens configuration, which consists completely of commercial singlets. The home-built objective has a numerical aperture (NA) of 0.44 and a long working distance of 35.9 mm, making it suitable for ex vacuo utilization. A diffraction-limited resolution of 1.08 µm and a field of view of about 210 µm are achieved when a 780 nm light passes through a 5 mm thick vacuum window. Moreover, such a design can be well adapted to a broad range of laser wavelengths (560-1000 nm) and vacuum window thicknesses (0-6 mm) by simply modifying one lens spacing, while maintaining a NA of above 0.43. The characteristics of the objective are evaluated experimentally, which are in good agreement with the simulations. Also, the objective has been successfully used for single-atom trapping and detecting in experiments. We believe that it will find more applications in various cold atom experiments.

[Diagnosis and Treatment of Epithelioid Hemangioendothelioma].

Objective To evaluate the clinical feature,diagnosis,and treatment of epithelioid hemangioendothelioma(EHE).Methods Data of 30 EHE patients admitted from January 2001 to December 2017 were retrospectively analyzed. The auxiliary examinations,treatment method,pathological findings,and prognoses were analyzed.Results There were 10 male patients and 20 females aged(47.9±16.1)years. The lesions sized(5.8±2.8)cm. Seventeen cases(56.7%)were asymptomatic and 17(56.7%)had no positive sign. The most common locations of EHE were scalp and spine,followed by liver. Nineteen patients underwent surgical operation(surgery group)and 11 only accepted needle biopsy(biopsy group). EHE was pathologically confirmed in all cases. CD34 and CD31 showed the highest positive rates in immunohistochemical stains. Twenty-three cases were followed up by(74.1±56.8)months. Seventeen cases survived and 6 died. The 1-,3-,and 5-year cumulative survival rates were 95.7%,86.3%,and 73.6%,respectively. The metastasis(14.3% vs. 77.8%,P=0.007)and mortality(7.1% vs. 55.6%,P=0.018)rates were significantly lower in surgery group than in biopsy group.Conclusions EHE is a rare tumor without specific symptom or sign. It can occur in any system of the body. Diagnosis depends mainly on pathology. The prognosis is acceptable. Complete surgical resection reduces distant metastases and mortality.

Cloning and Stress-Induced Expression Analysis of Calmodulin in the Antarctic Alga Chlamydomonas sp. ICE-L.

Calmodulin (CaM) is a Ca2+-binding protein that plays a role in several Ca2+ signaling pathways, which dynamically regulates the activities of hundreds of proteins. The ice alga Chlamydomonas sp. ICE-L, which has the ability to adapt to extreme polar conditions, is a crucial primary producer in Antarctic ecosystem. This study hypothesized that Cam helps the ICE-L to adapt to the fluctuating conditions in the polar environment. It first verified the overall length of Cam, through RT-PCR and RACE-PCR, based on partial Cam transcriptome library of ICE-L. Then, the nucleotide and predicted amino acid sequences were, respectively, analyzed by various bioinformatics approaches to gain more insights into the computed physicochemical properties of the CaM. Potential involvements of Cam in responding to certain stimuli (i.e., UVB radiation, high salinity, and temperature) were investigated by differential expression, measuring its transcription levels by means of quantitative RT-PCR. Results showed that CaM was indeed inducible and regulated by high UVB radiation, high salinity, and nonoptimal temperature conditions. Different conditions had different expression tendencies, which provided an important basis for investigating the adaptation mechanism of Cam in ICE-L.

Nanoporous Gyroid-Structured Epoxy from Block Copolymer Templates for High Protein Adsorbability.

Nanoporous epoxy with gyroid texture is fabricated by using a nanoporous polymer with gyroid-forming nanochannels as a template for polymerization of epoxy. The nanoporous polymer template is obtained from the self-assembly of degradable block copolymer, polystyrene-b-poly(l-lactide) (PS-PLLA), followed by hydrolysis of PLLA blocks. Templated polymerization can be conducted under ambient conditions to create well-defined, bicontinuous epoxy networks in a PS matrix. By taking advantage of multistep curing of epoxy, well-ordered robust nanoporous epoxy can be obtained after removal of PS template, giving robust porous materials. The through-hole nanoporous epoxy in the film state can be used as a coated layer to enhance the adsorbability for both lysozyme and bovine serum albumin.

Detection of Circulating Tumor Cells Using a Novel Immunomagnetic Bead Method in Lung Cancer Patients.

BACKGROUND: Circulating tumor cells (CTCs) are detectable in peripheral blood of metastatic lung cancer patients. In this article, we evaluate a new CTC separation method based on a combination of anti-EpCAM and immunomagnetic beads with the aim to detect CTCs more conveniently and specifically. METHODS: Lung cancer cells were magnetically labeled by anti-EpCAM magnetic beads, and subsequently captured by magnetic separation using our novel device. Isolated lung cancer cells were identified by pathomorphological by hematoxylin-eosin staining protocol. The system was used to detect CTCs in 2 ml blood. Blood samples of healthy donors spiked with lung cancer cell line A549 cells were used to determine the sensitivity and specificity of the method. Prevalence of CTCs was examined in samples from 56 patients with lung cancer. RESULTS: Regression analysis of number of recovered versus spiked A549 cells yielded a coefficient of determination of R(2) = 0.996 (P < 0.001). The average recovery was 68% or more at each spiking level. The coefficient of variation increased as the number of spiked cells decreased, ranging from 6.4% (1,000-cell spike) to 18.4% (50-cell spike). Forty-nine of the fifty-six patients (87.5%) were found to have CTCs in peripheral blood. None of the 2 ml peripheral blood samples of the 20 healthy subjects analyzed were found to have CTCs. CONCLUSIONS: This novel turbulence device provides a new tool allowing for feasible and specific detection of CTCs in lung cancer patients. It is likely clinically useful in diagnosis and monitoring of lung cancer and may have a role in clinical decision making.

Dimethyl phenyl piperazine iodide (DMPP) induces glioma regression by inhibiting angiogenesis.

1,1-Dimethyl-4-phenyl piperazine iodide (DMPP) is a synthetic nicotinic acetylcholine receptor (nAChR) agonist that could reduce airway inflammation. In this study, we demonstrated that DMPP could dramatically inhibit glioma size maintained on the chick embryonic chorioallantoic membrane (CAM). We first performed MTT and BrdU incorporation experiments on U87 glioma cells in vitro to understand the mechanism involved. We established that DMPP did not significantly affect U87 cell proliferation and survival. We speculated that DMPP directly caused the tumor to regress by affecting the vasculature in and around the implanted tumor on our chick CAM model. Hence, we conducted detailed analysis of DMPP's inhibitory effects on angiogenesis. Three vasculogenesis and angiogenesis in vivo models were used in the study which included (1) early chick blood islands formation, (2) chick yolk-sac membrane (YSW) and (3) CAM models. The results revealed that DMPP directly suppressed all developmental stages involved in vasculogenesis and angiogenesis - possibly by acting through Ang-1 and HIF-2α signaling. In sum, our results show that DMPP could induce glioma regression grown on CAM by inhibiting vasculogenesis and angiogenesis.

Mixed large cell neuroendocrine carcinoma and adenocarcinoma of the gallbladder: a case report and brief review of the literature.

Large-cell neuroendocrine carcinoma (LCNEC) of the gallbladder is extremely rare. We present a 63-year-old Chinese female who was admitted with right upper quadrant pain and a quasi-circular tumor measuring 2.0 cm on the body of the gallbladder, as indicated by computed tomography. LCNEC combined with adenocarcinoma was immunohistochemically confirmed after open radical cholecystectomy. Postoperative recovery of this patient was uneventful, and no evidence of recurrence or metastasis was observed after 12 months of follow-up. LCNEC of the gallbladder is thought to be extremely rare and is usually found in combination with other histological carcinoma types, such as adenocarcinoma, as determined histologically. The prognosis is poor overall, but early detection with complete resection may result in a relatively good prognosis.

The risk of carcinogenesis in congenital choledochal cyst patients: an analysis of 214 cases.

BACKGROUND: Choledochal cysts are most common in Asian populations. In addition, the incidence of biliary tract malignancies from choledochal cysts is increasing, but the risk of carcinogenesis is still unclear. MATERIAL AND METHODS: Clinical data from 214 congenital choledochal cyst cases from 1968 to 2013 were retrospectively analyzed. RESULTS: Todani type I was more common (139, 65.0%) than type IVa (53, 24.8%) or type V (17, 7.9%) in these choledochal cyst patients. Biliary tract malignant tumors occurred in the gallbladder (2, 13.3%), common bile duct (10, 66.7%), and intrahepatic bile duct (3, 20%) in 15 patients (7.0%), including one patient in whom malignant transformation occurred in the intrahepatic bile duct in a type IVa patient 15 years after extrahepatic cyst resection. An age at symptom onset ≥ 60 years was a risk factor (p < 0.001), while an initial complete surgery was a protective factor for carcinogenesis (p = 0.001). CONCLUSIONS: Choledochal cysts should be removed once diagnosed because of an increased risk of malignant transformation with increasing age. Complete cyst removal is necessary for the first surgical treatment. Additional hepatectomy should be considered for type IVa choledochal cysts because cholangiocarcinoma can arise from the intrahepatic bile duct years after the extrahepatic cyst excision.

The Research of No-Touch Isolation Technique on the Prevention of Postoperative Recurrence and Metastasis of Hepatocellular Carcinoma after Hepatectomy.

BACKGROUND/AIMS: To evaluate the efficacy of a surgical no-touch isolation technique in primary hepatocellular carcinoma patients compared with traditional hepatectomy. METHODOLOGY: Eighty hepatocellular carcinoma patients were randomly divided into traditional hepatectomy (n = 40) and no-touch isolation technique groups (n = 40). We compared peripheral blood variations in alpha-fetoprotein mRNA, miRNA-221, miRNA-224, and miRNA-122 expression levels in both groups pre- and postoperatively using real-time fluorescent quantitative polymerase chain reaction. RESULTS: The patients in the no-touch isolation technique group had better clinical curative effect. In the traditional hepatectomy group, variations in alpha-fetoprotein mRNA copy number pre- and postoperatively indirectly indicated a significant increase in the number of exfoliated carcinoma cells induced by manipulating the liver, increasing the risk of postoperative recurrence and metastasis (P < 0.05). Traditional hepatectomy patients showed higher increases in miRNA-221 and miRNA-224 expression than those in the no-touch isolation technique group (P < 0.05). Tumor resection resulted in preoperative expression and high postoperative expression of miRNA-122. No-touch isolation technique patients showed a slight and significant increase (P < 0.01) in miRNA-122, which was positively correlated with postoperative liver function index. Conclusion: The no-touch isolation technique is more effective than traditional hepa- tectomy in tumor resection for primary hepatocellular carcinoma.

Laparoscopic spleen-preserving total pancreatectomy for the treatment of low-grade malignant pancreatic tumors: Two case reports and review of literature.

BACKGROUND: Function-preserving pancreatectomy can improve the long-term quality of life of patients with benign or low-grade malignant tumors, such as intraductal papillary mucinous neoplasms (IPMNs) and mucinous cystic neoplasms. However, there is limited literature on laparoscopic spleen-preserving total pancreatectomy (L-SpTP) due to technical difficulties. CASE SUMMARY: Patient 1 was a 51-year-old male diagnosed with IPMN based on preoperative imaging, showing solid nodules in the pancreatic head and diffuse dilation of the main pancreatic duct with atrophy of the distal pancreas. We performed L-SpTP with preservation of the splenic vessels, and the postoperative pathology report revealed IPMN with invasive carcinoma. Patient 2 was a 60-year-old male with multiple cystic lesions in the pancreatic head and body. L-SpTP was performed, and intraoperatively, the splenic vein was injured and required ligation. Postoperative pathology revealed a mucinous cystic tumor of the pancreas with low-grade dysplasia. Both patients were discharged on postoperative day 7, and there were no major complications during the perioperative period. CONCLUSION: We believe that L-SpTP is a safe and feasible treatment for low-grade malignant pancreatic tumors, but more case studies are needed to evaluate its safety, efficacy, and long-term outcomes.

Glucometabolic-Related Genes as Diagnostic Biomarkers and Therapeutic Targets for Alzheimer's Disease and Type 2 Diabetes Mellitus: A Bioinformatics Analysis.

Background: Alzheimer's disease (AD) and type 2 diabetes mellitus (T2DM) are two widespread chronic disorders characterized by shared risk factors and molecular pathways. Glucose metabolism, pivotal for cellular homeostasis and energy supply, plays a critical role in these diseases. Its disturbance has been linked to the pathogenesis of both AD and T2DM. However, a comprehensive investigation into the specific roles of glucometabolic genes in the onset and progression of AD and T2DM has yet to be conducted. Methods: By analyzing microarray datasets from the Gene Expression Omnibus (GEO) repository, we identified differentially expressed glucometabolic genes (DEGs) in AD and T2DM cohorts. A range of bioinformatics tools were employed for functional annotation, pathway enrichment, protein interaction network construction, module analysis, ROC curve assessment, correlation matrix construction, gene set enrichment analysis, and gene-drug interaction mapping of these DEGs. Key genes were further validated using quantitative real-time polymerase chain reaction (qRT-PCR) in AD and T2DM murine models. Results: Our investigation identified 41 glucometabolic-related DEGs, with six prominent genes (G6PD, PKM, ENO3, PFKL, PGD, and TALDO1) being common in both AD and T2DM cohorts. These genes play crucial roles in metabolic pathways including glycolysis, pentose phosphate pathway, and amino sugar metabolism. Their diagnostic potential was highlighted by area under curve (AUC) values exceeding 0.6 for AD and 0.8 for T2DM. Further analysis explored the interactions, pathway enrichments, regulatory mechanisms, and potential drug interactions of these key genes. In the AD murine model, quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed significant upregulation of G6pd, Eno3, and Taldo1. Similarly, in the T2DM murine model, elevated expression levels of G6pd, Pfkl, Eno3, and Pgd were observed. Conclusion: Our rigorous research sheds light on the molecular interconnections between AD and T2DM from a glucometabolic perspective, revealing new opportunities for pharmacological innovation and therapeutic approaches. This study appears to be the first to extensively investigate glucometabolic-associated DEGs and key genes in both AD and T2DM, utilizing multiple datasets. These insights are set to enhance our understanding of the complex pathophysiology underlying these widespread chronic diseases.

Sclerosing epithelioid fibrosarcoma of the pancreas: A case report.

BACKGROUND: A sclerosing epithelioid fibrosarcoma (SEF) is a rare malignant fibroblastic soft tissue tumor that rarely occurs in intra-abdominal organs. A case of a SEF in the pancreatic head is reported herein, including its clinical manifestations, preoperative imaging features, gross specimen and pathological findings. CASE SUMMARY: A 33-year-old male patient was admitted to Peking Union Medical College Hospital in December 2023 due to a one-year history of intermittent upper abdominal pain and the discovery of a pancreatic mass. The patient underwent an enhanced computed tomography scan of the abdomen, which revealed a well-defined, round mass with clear borders and calcifications in the pancreatic head. The mass exhibited progressive, uneven mild enhancement, measuring approximately 6.6 cm × 6.3 cm. The patient underwent laparoscopic pylorus-preserving pancreaticoduodenectomy. Postoperative pathological examination revealed that the lesion was consistent with a SEF. At the 3-month postoperative follow-up, the patient did not report any short-term complications, and there were no signs of tumor recurrence. CONCLUSION: SEFs are rare malignant fibrous soft tissue tumors. SEFs rarely develop in the pancreas, and its preoperative diagnosis depends on imaging findings, with confirmation depending on pathological examination and immunohistochemistry. Currently, only four cases of pancreatic SEF have been reported in studies written in English. This case is the first reported case of a pancreatic SEF by a clinical physician.

Mirizzi syndrome: our experience with 27 cases in PUMC Hospital.

OBJECTIVE: To retrospectively evaluate the diagnosis and treatment of Mirizzi syndrome (MS). METHODS: Patients who received elective or emergency cholecystectomies in our center during 23 years were retrospectively evaluated. The data reviewed included demography, clinical presentations, diagnostic methods, surgical procedures, postoperative complications, and follow-up. RESULTS: There were 27 patients diagnosed with MS among 8697 cholecystectomies performed during that period. The preoperative diagnostic modalities included ultrasonography, computed tomography, magnetic resonance cholangiopancreatography, and endoscopic retrograde cholangiopancreatography. The incidence of MS Type I (12/27, 44.4%) had the dominance in the four types, the incidence of MS Type II and III were 33.3% (9/27) and 22.2% (6/27), and there were no MS Type IV patients. Laparoscopic cholecystectomy was performed in 15 (55.6%) patients, but only 3 (11.1%) patients with MS Type I had a successful surgery, and the other 12 were converted to open cholecystectomy. The remaining 12 patients directly underwent open cholecystectomy. The surgical procedures except laparoscopic cholecystectomy included simply open cholecystectomy (including laparoscopic cholecystectomy converted to open cholecystectomy) (6/27, 22.2%), open cholecystectomy, T-tube placement with choledochotomy (9/27, 33.3%), open cholecystectomy, closure of the fistula with gallbladder cuff, T-tube placement (3/27, 11.1%), and open cholecystectomy with excision of the external bile ducts, and Roux-en-Y hepatico-jejunostomy (6/27, 22.2%). Of them, 88.9% (24/27) patients recovered uneventfully and were discharged in good condition without any operation related mortality. CONCLUSIONS: Endoscopic retrograde cholangiopancreatography is a good method with diagnostic and therapeutic purposes. Total or partial cholecystectomy is generally adequate for MS Type I. For MS Type II-IV, paritial cholecystectomy, choledochoplasty, or if impossible, Roux-en-Y hepatico-jejunostomy may be performed. Laparoscopic cholecystectomy may be successful in selected preoperatively diagnosed MS Type I patients, and open cholecystectomy is the standard therapeutic method.

Expression of microRNA-29b2-c cluster is positively regulated by MyoD in L6 cells.

OBJECTIVES: To evaluate the expression profile of myoD microRNA-29 (miR-29) family in L6 myoblast differentiated to myotube of L6 myotube treated by glucose and insulin, and to further probe the molecular mechanism of myoD regulating the expression of miR-29 clusters. METHODS: The expression of myoD and miR-29 family was detected by using real-time PCR and Western blot analysis. The potential promoter and transcription factors binding sites of miR-29 clusters were predicted by Promoter scan and transcriptional factor search. The promoter sequence of miR-29b1-a and miR-29b2-c cluster was cloned into a luciferase reporter plasmid and the regulatory effect of myoD was analyzed by using dual luciferase reporter assay. Electrophoretic mobility shift assay was further conducted to indicate the binding of myoD on specific sequence. Moreover, overexpression of myoD was achieved by a recombinant adenovirus system (Ad-myoD). L6 cells were infected with Ad-myoD and real-time PCR was conducted to analyze the expression of miR-29b and miR-29c. RESULTS: The expression levels of myoD, miR-29a, miR-29b, and miR-29c were increased in L6 myoblast differentiated to myotube. The expression of myoD, miR-29b, and miR-29c was up-regulated in L6 myotube treated with glucose and insulin, but miR-29a depicted no significant change. Dual luciferase reporter gene assay showed that myoD functioned as a positive regulator of miR-29b2-c expression and myoD could bind to the specific sequence located at the promoter region of miR-29b2-c cluster. Enforced expression of myoD led to a marked increase of miR-29b and miR-29c levels in L6 cells. CONCLUSION: MyoD might act as a crucial regulator of myogenesis and glucose metabolism in muscle through regulating the expression of miR-29b2-c.

[Construction of RGD10-NGR9 dual-targeting superparamagnetic iron oxide and its magnetic resonance imaging features in nude mice].

OBJECTIVE: To construct angiogenesis-specific RGD10-NGR9 dual-targeting superparamagnetic iron oxide nanoparticles, and to evaluate its magnetic resonamce imaging (MRI) features in nude mice and potential diagnostic value in tumor MRI. METHODS: Dual-targeting peptides RGD10-NGR9 were designed and synthesized. Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles were synthesized by chemical co-precipitation method and the surface was modified to be hydrophilic by coating with dextran. The dual-targeting peptides RGD10-NGR9 were conjugated to USPIO. Cell binding affinity and up-taking ability of the dual-targeting USPIO nanoparticles to integrin ανß3-APN positive cells were subsequently tested by Prussian blue staining and phenanthroline colorimetry in vitro. The RGD10-NGR9 conjugated with USPIO was injected intravenously into xenograft mice, which were scanned by MRI at predetermined time points. The MRI and contrast-to-noise ratio (CNR) values were calculated to evaluate the ability of dual-targeting USPIO as a potential contrast agent in nude mice. RESULTS: P-CLN-Dextran-USPIO nanoparticles with stable physical properties were successfully constructed. The average diameter of Fe3O4 nanoparticles was 8-10 nm, that of Dextran-USPIO was about 20 nm and P-CLN-Dextran-USPIO had an average diameter about 30 nm. The in vitro studies showed a better specificity of dual-targeting USPIO nanoparticles on proliferating human umbilical vein endothelia cells (HUVEC). In vivo, RGD10-NGR9-USPIO showed a significantly reduced contrast in signal intensity and 2.83-times increased the CNR in the tumor MRI in xenograft mice. CONCLUSION: This novel synthesized RGD10-NGR9 dual-targeting USPIO is with better specific affinity in vitro and in vivo, and might be used as a molecular contrast agent for tumor angiogenesis MRI.