The Progress on Magnetic Material Thin Films Prepared Using Polymer-Assisted Deposition.

Polymer-assisted deposition (PAD) has been widely used in the preparation of high-quality oxides and sulfides for basic research and applications. Specifically, diverse PAD-prepared magnetic material thin films such as ZnO, Ga2O3, SrRuO3, LaCoO3, LaMnO3, Y3Fe5O12, MoS2, MoSe2, and ReS2 thin films have been grown, in which thickness-dependent, strain-modulated, doping-mediated, and/or morphology-dependent room-temperature ferromagnetism (RTFM) have been explored. Inspired by the discovery of intrinsic low-temperature FM in two-dimensional (2D) systems prepared using mechanical exfoliation, the search for more convenient methods to prepare 2D ferromagnetic materials with high-temperature FM has seen explosive growth, but with little success. Fortunately, the very recent synthesis of 2D NiO by PAD has shed light on this challenge. Based on these abovementioned developments, the difficulties of PAD when preparing a-few-nanometer single-crystalline materials and the opportunities in PAD for novel materials such as chiral magnetic soliton material Cr1/3NbS2 are discussed.

Toxicologic evaluation of repetitive 4-week intravenous injections of midkine antisense oligonucleotide nanoliposomes in rats.

Midkine antisense oligonucleotide (MK-ASODN) nanoliposomes have previously been shown to have inhibitory activity against hepatocellular carcinoma growth. Herein we report the 4-week sub-chronic toxicity of MK-ASODN nanoliposomes in SD rats. The adverse effects included loss of body weight gain and food consumption, peri-rhinal bleeding, piloerection, peri-anal filth, and kidney, liver, spleen, thymus, lung, and injection site lesions at high doses. Macroscopic changes were observed in the kidneys of the high-dose group, accompanied by a variation in urine protein and white blood cells, blood urea nitrogen, and serum creatinine. The increased spleen and liver coefficient, and the variation in circulating white blood cells, lymphocytes, and eosinophils in the high-dose group demonstrated that inflammation was caused by MK-ASODN nanoliposomes and was consistent with the macroscopic changes in the spleen and liver. The main necropsy findings of the animals that died were macroscopic changes in the lung. No severe toxic effects or mortalities occurred in the low- and medium-dose groups. However, a No Adverse Effect Level (NOAEL) was not identified since there were changes in organs deemed to be adverse at all dose levels. Thus, the maximum tolerated dose of MK-ASODN nanoliposomes for rats was considered to be 6 mg/kg/day.

Photocurable and elastic polyurethane based on polyether glycol with adjustable hardness for 3D printing customized flatfoot orthosis.

Orthopedic insoles is the most commonly used nonsurgical treatment method for the flatfoot. Polyurethane (PU) plays a crucial role in the manufacturing of orthopedic insoles due to its high wear resistance and elastic recovery. However, preparing orthopedic insoles with adjustable hardness, high-accuracy, and matches the plantar morphology is challenging. Herein, a liquid crystal display (LCD) three-dimensional (3D) printer was used to prepare the customized arch-support insoles based on photo-curable and elastic polyurethane acrylate (PUA) composite resins. Two kinds of photo-curable polyurethanes (DL1000-PUA and DL2000-PUA) were successfully synthesized, and a series of fast-photocuring polyurethane acrylate (PUA) composite resins for photo-polymerization 3D printing were developed. The effects of different acrylate monomers on the Shore hardness, viscosity, and mechanical properties of the PUA composite resins were evaluated. The PUA-3-1 composite resin exhibited low viscosity, optimal hardness, and mechanical properties. A deviation analysis was conducted to assess the accuracy of printed insole. Furthermore, the stress conditions of the PUA composite resin and ethylene vinyl acetate (EVA) under the weight load of healthy adults were compared by finite element analysis (FEA) simulation. The results demonstrated that the stress of the PUA composite resin and EVA were 0.152 MPa and 0.285 MPa, and displacement were 0.051 mm and 3.449 mm, respectively. These results indicate that 3D-printed arch-support insole based on photocurable PUA composite resin are high-accuracy, and can reduce plantar pressure and prevent insoles premature deformation, which show great potential in the physiotherapeutic intervention for foot disorders.

Recycling of waste crab shells into reinforced poly (lactic acid) biocomposites for 3D printing.

To promote natural waste resource utilization, a novel biocomposite, composed of waste crab shells and poly (lactic acid) matrix, was developed by combining chemical treatment and 3D printing. A crab shell powder (ISCSP) with an abundant porous structure and a high specific surface area was obtained by treatment with hydrochloric acid and sodium hydroxide. Importantly, under the optimal printing parameters determined by the finite element analysis, test samples, and porous bones were successfully printed using CSP/PLA composites by a commercial fused deposition modeling (FDM) 3D printer. The morphology, mechanical and thermal properties, antibacterial properties, and biocompatibility of the CSP/PLA composites were then assessed. Our results revealed that the tensile strength and flexural strength of the ISCSP/PLA composites reached 58.71 and 90.11 MPa, which were 28.6 % and 28.8 % higher than that of pure PLA, respectively. The glass transition and melting temperatures of the composites remained similar to those of pure PLA. Interestingly, the addition of CSP increased PLA crystallinity, which could be attributed to the nucleation effect of CSP in the system. The antibacterial activity of the PLA-1.5ESCSP composite samples against Escherichia coli (E. coli) was greater than 99 %. More importantly, the live/dead assay showed that the CSP/PLA composites possessed excellent biocompatibility. Therefore, the developed CSP/PLA biocomposites are potential feedstocks for 3D printing in bone tissue engineering and may be used as graft substitutes in reparative and reconstructive surgery. They are especially beneficial due to their superior mechanical and thermal properties, excellent antibacterial activities, and significant biocompatibility.

3D bio-printing of photocrosslinked anatomically tooth-shaped scaffolds for alveolar ridge preservation after tooth extraction.

Alveolar ridge preservation techniques have been developed as a possible method to maintain the optimum ridge contour and dimensions. Grafting a bone substitute is paramount to prevent alveolar ridge resorption after tooth extraction. However, it remains a great challenge to develop alveolar ridge preservation materials with sufficient mechanical strength, bioactivity, and osteoinductivity and favorable tooth extraction socket morphological matching. In this work, a novel photocrosslinked composite ink consisting of nacre, polyurethane (PU) and polyhedral oligomeric silsesquioxane (POSS) was prepared and used to fabricate 3D porous scaffolds for alveolar ridge preservation. This nacre/PU/POSS (NPP) composite was characterized in terms of its rheological behavior, mechanical properties, and surface hydrophilicity. The biomineralization of these NPP scaffolds was confirmed via in vitro experiments. MC3T3-E1 cells were distributed homogeneously on the NPP scaffolds and stimulated cellular proliferation. When the NPP scaffolds were grafted into the sockets after extraction of mandibular incisors, the height and width of alveolar bone resorption were reduced, and new bone formation was observed. These NPP composites are promising scaffold materials for alveolar ridge preservation and 3D printing of bone grafts in future.

Scarless Healing of Injured Vocal Folds Using an Injectable Hyaluronic Acid-Waterborne Polyurethane Hybrid Hydrogel to Tune Inflammation and Collagen Deposition.

Vocal fold (VF) scarring results from injury to the unique layered structure and is one of the main reasons for long-lasting dysphonia. A minimally invasive procedure with injectable hydrogels is a promising method for therapy. However, current surgical techniques or standard injectable fillers do not yield satisfactory outcomes. In this work, an injectable hybrid hydrogel consisting of oxide hyaluronic acid and hydrazide-modified waterborne polyurethane emulsion was injected precisely into the injury site and cross-linked in situ by a dynamic hydrazone bond. The prepared hydrogel displays excellent injectability and self-healing ability, showing favorable biocompatibility and biodegradability to facilitate endogenous newborn cell migration and growth for tissue regeneration. With the aim of evaluating the antifibrosis and regeneration capacity of the hybrid hydrogel in the VF scarring model, the morphology and vibration characteristics of VFs, inflammatory response, and healing status were collected. The hybrid hydrogel can decrease the inflammation and increase the ratio of collagen III/collagen I to heal damaged scar-free tissue. Fascinatingly, the mucosal wave oscillations of healing VF by injecting the hybrid hydrogel were vibrated like the normal VF, achieving functional restoration. This work highlights the utility of hybrid hydrogels consisting of synthetic biodegradable waterborne polyurethane emulsions and natural hyaluronic acid as promising biomaterials for scarless healing of damaged VFs.

Comparison of fibrin glue versus suture for conjunctival autografting in pterygium surgery: a meta-analysis.

PURPOSE: To evaluate the safety and clinical efficacy of fibrin glue in pterygium surgery with conjunctival autografting. DESIGN: The use of fibrin glue has been introduced in the treatment of pterygium. However, its role versus traditional suturing is still a matter of debate. We performed a meta-analysis to compare the safety and clinical efficacy of fibrin glue with suture for conjunctival autograft attachment in pterygium surgery. PARTICIPANTS: A total of 342 participants with 366 eyes in 7 studies were analyzed. METHODS: We searched Medline, EMBASE, Web of Science, Cochrane Central Register of Controlled Trials, and Google Scholar for relevant randomized controlled trials (RCTs). MAIN OUTCOME MEASURES: The methodological quality of all the included trials was assessed with the Jadad score. The meta-analysis was performed with the fixed-effects model for complication rate and recurrence rate, and random-effects model for operating time. RESULTS: Fibrin glue was associated with a significantly decreased operating time (weighted mean difference -17.61 minutes, 95% confidence interval [CI], -26.03 to -9.18, P<0.0001) and was more effective in reducing the recurrence rate (Peto odds ratio [OR] 0.33, 95% CI, 0.15-0.71, P = 0.004) compared with suture. There were no significant differences in the complication rate (Peto OR 1.82, 95% CI, 0.63-5.27, P = 0.27) between the 2 groups. CONCLUSIONS: Our meta-analysis supports the superiority of fibrin glue to suture in pterygium surgery with conjunctival autografting in that the use of fibrin glue can significantly reduce the recurrence rate without increasing the risk of complications. Ophthalmologists should consider the use of fibrin glue in pterygium surgery. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Toward Rapid and Sensitive Detection of SARS-CoV-2 with Functionalized Magnetic Nanoparticles.

The outbreak of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) threatens global medical systems and economies and rules our daily living life. Controlling the outbreak of SARS-CoV-2 has become one of the most important and urgent strategies throughout the whole world. As of October 2020, there have not yet been any medicines or therapies to be effective against SARS-CoV-2. Thus, rapid and sensitive diagnostics is the most important measures to control the outbreak of SARS-CoV-2. Homogeneous biosensing based on magnetic nanoparticles (MNPs) is one of the most promising approaches for rapid and highly sensitive detection of biomolecules. This paper proposes an approach for rapid and sensitive detection of SARS-CoV-2 with functionalized MNPs via the measurement of their magnetic response in an ac magnetic field. For proof of concept, mimic SARS-CoV-2 consisting of spike proteins and polystyrene beads are used for experiments. Experimental results demonstrate that the proposed approach allows the rapid detection of mimic SARS-CoV-2 with a limit of detection of 0.084 nM (5.9 fmole). The proposed approach has great potential for designing a low-cost and point-of-care device for rapid and sensitive diagnostics of SARS-CoV-2.

SPECT imaging and highly efficient therapy of rheumatoid arthritis based on hyperbranched semiconducting polymer nanoparticles.

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease. Although significant progress has been made in clinical treatment, joint inflammation may continue or worsen, and may even progress to the end-stage that requires joint replacement. Traditional therapy using methotrexate (MTX) would cause serious off-target systemic toxicities. Therefore, it is crucial to effectively and specifically deliver MTX to targeted inflamed joints to decrease its adverse systemic toxicities and improve its therapeutic index. Herein, we develop multifunctional nanocarriers for diagnostic radioisotope (99mTc) labeling and therapeutic targeted drug (MTX) delivery by using PEGylated hyperbranched semiconducting polymer nanoparticles (HSP-PEG-NPs) as carriers. Upon intravenous administration, the nanoparticles can extravasate through the turbulent blood-joint barrier and access the inflamed joints. In vivo SPECT/CT imaging shows high accumulation in the inflamed joints of mice with RA after intravenous injection of HSP-PEG-NPs with 99mTc labeling (99mTc-HSP-PEG). In vivo therapeutic evaluations suggest that MTX@HSP-PEG-NPs significantly alleviate RA with a high therapeutic index and relatively low adverse systemic toxicities in comparison with free MTX at the same dose. Our study shows that HSP-PEG-NPs could serve as multifunctional vehicles to deliver radioisotopes for in vivo imaging, and MTX for RA treatment, highlighting the innovative development of the nanoparticle-based RA treatment strategy for clinical applications.

Study on the anti-infection ability of vancomycin cationic liposome combined with polylactide fracture internal fixator.

A polylactide composite fracture fixator loaded with vancomycin cationic liposome (PLA@VL) was prepared by reverse evaporation method. The method of cationic liposome encapsulating vancomycin could effectively improve antibacterial property and achieve drug sustained release effect, so as to reduce toxicity of antibiotics in vivo. Scanning electron microscope (SEM) was used to observe morphology and Fourier transform infrared spectroscopy (FTIR) was used to detect the composition of the internal fixator. In vitro drug release model, in vitro degradation model and body fluid osteogenesis model were designed in this study. On the other hand, the experiments of inhibition zone and MC3T3-E1 osteoblasts in mice were conducted to explore antibacterial property, cell activity and adhesion of the PLA@VL composite internal fixator. Alkaline phosphatase (ALP) staining method and alizarin red assay were used to detect the osteogenic induction ability of the composite internal fixator. Finally, mice fracture models were established to verify osteogenic and anti-infection abilities of the composite internal fixator in vivo. The results showed that MC3T3-E1 cells had better adhesion and proliferation abilities on the PLA@VL composite internal fixator than on the PLA fixator, which indicated that the PLA@VL composite internal fixator possessed excellent osteogenic and anti-infection abilities both in vivo and in vitro. Therefore, the above experiments showed that the fracture internal fixator combined with vancomycin cationic liposome had better biocompatibility, antibacterial ability and osteogenic ability, which provides a promising anti-infection material for the clinical field of fracture.

Attenuated Salmonella typhimurium-mediated tumour targeting imaging based on peptides.

Attenuated bacteria-mediated tumor targeting diagnosis will provide a novel strategy for further cancer treatments owing to the intrinsic facultative anaerobic characteristic of bacteria and rapid proliferation in the tumor sites. In this work, we firstly investigate the in vivo behaviour of the attenuated Salmonella typhimurium (S. typhimuriumΔppGpp/lux) after intravenous injection. S. typhimurium exhibits rapid proliferation in tumor sites after three days of injection through bioluminescence imaging, the Luria-Bertani plate and the Gram-staining assay. In contrast, S. typhimurium does not proliferate in the normal tissues and could be excreted from the body of mice. Afterwards, a targeting peptide ubiquicidin (UBI) labeled with fluorescent dye Cy5.5 or radionuclide 125I was intravenously injected into the mice with or without S. typhimurium treatments for in vivo fluorescence imaging and single-photon emission computed tomography (SPECT/CT) imaging, respectively. The results show that the peptide UBI could specifically target the two independent bacteria-infected tumor models, the 4T1 murine breast cancer model and the CT26 mouse colon cancer model, realizing the sensitive multimodal imaging of tumors. Such a strategy (bacteria-mediated tumor targeting) may further improve the sensitivity to early diagnosis of tumors. We hope that our developed strategy could further be extended to cancer theranostics in the future, bringing good news for cancer patients.

Evaluation of the Safety and Efficacy of a Low-Temperature Plasma Surgical System for Pterygium.

PURPOSE: Surgical excision is the standard treatment for pterygium. This study was conducted to evaluate the safety and efficacy of a novel technique using low-temperature plasma (LTP) for excision and hemostasis in pterygium surgery. METHODS: A prospective, comparative, and randomized clinical trial was conducted on 60 patients (60 eyes) undergoing pterygium excision with conjunctival autografts using fibrin glue. Patients were equally divided into the following 2 groups: a control group and a LTP group. Postoperative follow-up visits were scheduled on day 1, week 1, and months 1 and 3, and recurrence was evaluated at 1 year. Patients were examined for operative time, best corrected visual acuity, conjunctival autograft inflammation (CAI), graft stability (GS), pain, recurrence, and final appearance. Factors related to pterygium recurrence and final appearance were analyzed. RESULTS: Mean operative times were shorter in the LTP group (16.7 ± 3.4 min) than those in the control group (20.1 ± 4.7 min, P = 0.002). LTP eyes had milder CAI than control eyes at postoperative day 1 (P = 0.000) and week 1 (P = 0.000). Patients in the LTP group exhibited better GS (P = 0.01) and milder pain (P = 0.04) than those in the control group on day 1. Two control patients (6.7%) and no (0%) LTP patients experienced recurrence (P = 0.08). GS and CAI were the significant factors contributing to recurrence (GS: R = 0.425, P = 0.001; CAI: R = 0.309, P = 0.016). CONCLUSIONS: LTP to replace surgical blades and disposable cautery for ablation and hemostasis is safe and efficient for pterygium surgery, resulting in shorter operative time, milder inflammation, and better graft stability without increasing complication risk.

Plant-derived chlorophyll derivative loaded liposomes for tri-model imaging guided photodynamic therapy.

Plant-derived chlorophyll derivatives with a porphyrin ring structure and intrinsic photosynthesis have been widely used in biomedicine for cancer theranostics. Owing to their poor water solubility, chlorophyll derivatives are very difficult to use in biomedical applications. In this work, pyropheophorbide acid (PPa) (liposome/PPa) nanoparticles, a liposome-encapsulated chlorophyll derivative, are designed for tri-model imaging guided photodynamic therapy (PDT) of cancer. The obtained liposome/PPa nanoparticles significantly enhance the water solubility of PPa, prolong blood circulation and optimize the bio-distribution in mice after intravenous injection. Utilizing their intrinsic fluorescence, high near-infrared (NIR) absorbance and extra radiolabeling, liposome/PPa nanoparticles could be used as excellent contrast agents for multimodal imaging including fluorescence (FL) imaging, photoacoustic (PA) imaging and SPECT/CT imaging. Under 690 nm laser irradiation at a low power density, liposome/PPa nanoparticles significantly inhibit tumor growth, further demonstrating the therapeutic efficiency of PDT using PPa. Therefore, our work developed liposome/PPa nanoparticles as multifunctional nanoagents for multimodal imaging guided PDT of cancer. This will further prompt the clinical applications of PPa in the future.

[Efficacy of Altemeier procedure in the treatment of rectal prolapse].

OBJECTIVE: To evaluate the safety and efficacy of the perineal rectosigmoidectomy (Altemeier procedure) in the treatment of full thickness rectal prolapse. METHODS: Clinical and follow-up data of 52 patients with full thickness rectal prolapse undergoing Altemeier procedure in 9 hospitals from September 2010 to July 2016 were analyzed retrospectively. Of which 38 cases were from Zhongnan Hospital of Wuhan University, 1 case from Suizhou Central Hospital of Hubei province, 1 case from Jingzhou Second People's Hospital of Hubei province, 3 cases from Wuxue First People's Hospital of Hubei province, 1 case from Jingmen First People's Hospital of Hubei province, 1 case from Tuanfeng County Hospital of Hubei province, 4 cases from Jingzhou Central Hospital of Hubei province, 2 from PLA Rocket Army General Hospital, 1 case from the Affiliated Hospital of Xuzhou Medical University in Jiangsu province. Altemeier operation steps: The line shaped teeth, the prolapsed rectum is first exposed to the anus. In the dentate line proximal 1-3 cm with ultrasonic knife or Ligasure ring outer rectal incision, using electric knife to mark pre resection line in rectal mucosa. Open down in front of the pelvic peritoneum. Incision of the outer intestine and the reduction of the internal rectum and part of sigmoid colon. To free and remove excess pelvic retroperitoneal, pelvic peritoneum and be at the top of the colon or rectum anterior pelvic reconstruction suture. The rear of the levator ani muscle forming rectum. Pull gently to the anus and rectum and sigmoid, in the absence of tension, 2-3 cm outside the anus was selected as the proximal inner bowel pre resection line, along the line of pre transection of proximal bowel resection, again the broken end of intestine full-thickness end-to-end anastomosis. Postoperative complication and recurrence were summarized. Gastrointestinal quality of life index (GIQLI), Wexner constipation score and Wexner fecal incontinence score were used to evaluate the efficacy. RESULTS: All the 52 patients were beyond moderate full thickness rectal prolapse. Thirty-one were male and 21 were female with age ranging from 22 to 83 (average 53) years. The length of prolapsed rectum was 6 to 20 (average 9) cm and course of disease was 0.5 to 46(average 19.5) years. No perioperative death. Five patients (9.6%) had postoperative complications, including 2 anastomotic bleeding, 1 wall portion dehiscence of anastomosis, 1 anastomotic stenosis, and 1 malnutrition. Recurrence rate was 9.6%(5/52) within the long-term follow-up of 5 to 71 (median 40) years. Compared with the preoperative results, Wexner constipation score and Wexner fecal incontinence score decreased obviously (2.1±1.4 vs. 4.6±3.4, 4.8±4.1 vs. 6.8±4.1), and GIQLI significantly increased from 99.6±8.0 to 103.0±9.1 (all P<0.05) at 6-month after operation. Above 3 scores were sustained and continuously improved at 12-, 24-, and 36-month during the follow-up (all P<0.05). CONCLUSION: Altemeier procedure possesses good efficacy with low morbidity of complication and recurrence in the treatment of full thickness rectal prolapse.

A Hydrogel-Based Hybrid Theranostic Contact Lens for Fungal Keratitis.

Fungal keratitis, a severe ocular disease, is one of the leading causes of ocular morbidity and blindness, yet it is often neglected, especially in developing countries. Therapeutic efficacy of traditional treatment such as eye drops is very limited due to poor bioavailability, whereas intraocular injection might cause serious side effects. Herein, we designed and fabricated a hybrid hydrogel-based contact lens which comprises quaternized chitosan (HTCC), silver nanoparticles, and graphene oxide (GO) with a combination of antibacterial and antifungal functions. The hydrogel is cross-linked through electrostatic interactions between GO and HTCC, resulting in strong mechanical properties. Voriconazole (Vor), an antifungal drug, can be loaded onto GO which retains the drug and promotes its sustained release from the hydrogel-based contact lenses. The contact lenses also exhibited good antimicrobial functions in view of glycidyltrimethylammonium chloride and silver nanoparticles. The results from in vitro and in vivo experiments demonstrate that contact lenses loaded with Vor have excellent efficacy in antifungal activity in vitro and could significantly enhance the therapeutic effects on a fungus-infected mouse model. The results indicate that this hydrogel contact lenses-based drug delivery system might be a promising therapeutic approach for a rapid and effective treatment of fungal keratitis.

Preparation and characterization of poly(triallyl isocyanurate-co-trimethylolpropane triacrylate) monolith and its applications in the separation of small molecules by liquid chromatography.

A new polymeric monolith was prepared in stainless-steel column and fused-silica capillary, respectively, by atom transfer radical polymerization technique. In the polymerization, triallyl isocyanurate (TAIC) was used as the functional monomer; trimethylolpropane triacrylate (TMPTA) as the crosslinking agent; polyethylene glycol 200 and 1,2-propanediol as the co-porogens; carbon tetrachloride as the initiator and ferrous chloride as the catalyst. The conditions of polymerization were optimized. Morphology of the prepared poly(TAIC-co-TMPTA) monolith was investigated by scanning electron microscopy; pore properties were assayed by mercury porosimetry and nitrogen adsorption. The characterization indicated that the prepared reversed-phase monolith possessed uniform structure, good permeability and mechanical stability. The column was used as the stationary phase of reversed phase high performance liquid chromatography (RP-HPLC) and capillary liquid chromatography (CLC) to separate the mixture of aromatic compounds. The new column performed around 125,000 theoretical plates per meter. The column showed good reproducibility: the relative standard deviation values of the retention factor values for aromatic compounds were less than 1.52% (n=7, column-to-column).

[Development of EV71, CA16 and other enterovirus vrial real-time qualitative PCR diagnostic kit].

OBJECTIVE: A novel multiplex real-time RT-PCR kit was developed to detect EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives, which used for hand, foot and mouth disease in the clinical diagnosis and epidemiological surveillance. METHODS: Design specific primers and probes of EV71, CA16, other intestinal virus and internal amplification control, improve the extraction method of virus nucleic acid. Optimization the detection system of real-time quantitative PCR. Research the products of the accuracy, stability, precision, amplification efficiency and detection of linear range. RESULTS: The primers and probes had high spicificity. The Viral RNA extraction effect of this Kit is as same as that of QIAamp Viral RNA mini Kit (QIAGEN company), but less reagent cost. The optimal concentrations of primers and probes are 0.2 micromol/L for all the upstream and downstream primers, 0.06 micromol/L for probes of other human enteroviruse, 0.08 micromol/L for probes of EV71 and CA16 respectively. The kit has good stability, accuracy and precision. The amplification efficiencies of EV71, CoxA16 and other human enteroviruses are 106% ,101% and 105% and the detection of linear range is from 10(9) copies/microl-10(2) copies/microl. CONCLUSION: The novel multiplex real-time RT-PCR kit for detecting EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control has good stability, accuracy, precision and amplification efficiencies. So it has great value in clinical application.

Large scale preparation of midkine antisense oligonucleotides nanoliposomes by a cross-flow injection technique combined with ultrafiltration and high-pressure extrusion procedures.

The midkine antisense oligonucleotide (MK-ASODN, 5'-CCC CGG GCC GCC CTT CTT CA-3') nanoliposomes have been identified to suppress hepatocellular carcinoma (HCC) growth effectively, and have a great potential to be an effective target drug for HCC. In this study, a facile and reproducible method for large-scale preparation of MK-ASODN nanoliposomes followed by lyophilization has been developed successfully. Meanwhile, the MK-ASODN nanoliposomes characteristics, storage stability and their antitumor efficiency were studied. The mean particle size of MK-ASODN nanoliposomes were 229.43±15.11 nm, and the zeta potential were 29.7±1.1 mV. High entrapment efficiency values were achieved around 90%. Transmission electron microscopy images revealed spherical shaped nanoliposomes. Nanoliposomes allowed sustained MK-ASODN release for as long as 14 days. During 180 days of storage, freeze-dried nanoliposomes showed no significant change in the mean size, zeta potential, entrapment efficiency and drug release ratio. Regarding their antitumor efficiency, the in vitro proliferation of human liver cancer cells were significantly inhibited by the MK-ASODN nanoliposomes. Furthermore, the MK-ASOND nanoliposomes also significantly inhibited the growth of HCC in the mouse model. In summary, the results confirmed that this large-scale preparation of MK-ASOND nanoliposomes was facile and reproducible, and potentially, could speed up the application process of our MK-ASOND nanoliposomes for HCC therapy.