Dendronized Hyperbranched Polyethyleneimines with Switchable Thermoresponsiveness and Encapsulation

Hyperbranched polyethyleneimines (PEIs) with multiple layered structures and high density of terminal amino groups have shown great potential for applications in biomedicine, sensors, and catalysis. In this work, a kind of dendronized hyperbranched PEIs (DHPs) is synthesized by modification of PEIs with dendritic oligoethylene glycols (OEGs). These DHPs not only inherit the characteristic thermoresponsive behavior from the OEG dendrons, but also show low cytotoxicity and switchable encapsulation capacities. Their thermoresponsive behavior is found to be mainly dependent on the coverage and amphiphilicity of dendritic OEGs, solution pH, and NaCl concentration. Moreover, DHPs exhibit advantages in thermally-switchable complexation with guest molecules because of their adjustable crowding effect from the densely packed OEG chains. In addition, these DHPs can also be used as nanoreactors, which can reduce metal ions into metal nanoparticles in a convenient way. In virtue of these peculiarities, it is believed that these dendronized hyperbranched PEIs with smart properties can be used as promising scaffolds in drug-controlled release and nanoreactors.     

Understanding peroral absorption: regulatory aspects and contemporary approaches to tackling solubility and permeability hurdles

Oral drug absorption is a process influenced by the physicochemical and biopharmaceutical properties of the drug and its inter-relationship with the gastrointestinal tract. Drug solubility, dissolution and permeability across intestinal barrier are the key parameters controlling absorption. This review provides an overview of the factors that affect drug absorption and the classification of a drug on the basis of solubility and permeability. The biopharmaceutical classification system (BCS) was introduced in early 90׳s and is a regulatory tool used to predict bioavailability problems associated with a new entity, thereby helping in the development of a drug product. Strategies to combat solubility and permeability issues are also discussed.     

限制性内切酶EcoRI的分离纯化

以大肠杆菌ＲＹ＿（１３），Ｋ＿（１２１１００）为酶源菌株，采用聚乙烯亚胺分离纯化了限制性内切酶ＥｃｏＲＩ，并对λ－ＤＮＡ有５个专一的切割位点，将λ－ＤＮＡ切割成６个分子量不同的片段，经琼脂糖凝胶电泳后，可得６个迁移位置不同的条带。这些特性使它特别适合于作为ＤＮＡ结构功能等方面研究的工具酶。在遗传工程研究中，限制酶更起了决定性作用。     

Degradation and resynthesis of cyclic 3', 5' -guanosine monophosphate in truncated rod photoreceptors from bovine retina

Truncated rod photoreceptors containing outer segments with inner segment attachments were prepared from bovine retinas. In the absence of a high-energy donor, the preparation transformed cyclic GMP to 5′-GMP, guanosine and a small amount of cyclic XMP. In the presence of ATP. the preparation formed GDP and GTP from 5′-GMP, and GTP endogenously formed was converted to cyclic GMP. Conditions for optimal activity of the above reactions were defined and a rank ordering of the specific activities of the reactions established. The specific activity of each enzyme was greater in truncated photoreceptors than in homogenates of retina. The efficiency of the metabolic cycle to resynthesize cyclic GMP from 5′-GMP in vivo probably depends upon the energy state of the visual cell and upon the degree of compartmentation of substrates within the inner and outer segment of the photoreceptor.     

RNA interference targeting hypoxia inducible factor 1alpha reduces post-operative adhesions in rats

BACKGROUND: To investigate the use of RNA interference mediated gene down-regulation targeting hypoxia inducible factor 1alpha (HIF-1alpha) and plasminogen activator inhibitor 1 (PAI-1) in an effort to prevent abdominal adhesion formation. MATERIALS AND METHODS: Real time PCR and a PAI-1 protein activity assay were used in vitro to determine the efficacy of small interfering RNAs (siRNAs). For in vivo experiments, 57 white female rats were operated to generate ischemic and serosal injury to the uterine horns, and treated with saline, siRNA(Lamin A/C) (negative control), siRNA(HIF-1alpha), siRNA(PAI-1), or siRNA(HIF-1alpha) plus siRNA(PAI-1). The cationic polyer poly(ethylenimine) (PEI) was used as the delivery vehicle for all siRNAs delivered in vivo. Adhesions were analyzed by a blinded surgeon 8 days post-surgery. RESULTS: After in vitro transfection with siRNA, at least 69% gene down-regulation was obtained for all siRNAs tested. In vitro siRNA-mediated down-regulation of HIF-1alpha, PAI-1 or their simultaneous delivery resulted in a significant decrease of PAI-1 protein activity (at least P < 0.05). Administration of 4 nmol siRNA(HIF-1alpha)/PEI complexes after injury to the uterine horns achieved a statistical reduction of post-operative adhesion formation with a reduction by 52% (P < 0.05). Delivery of 4 nmol siRNA(PAI-1)/PEI complexes and the simultaneous delivery of 2 nmol siRNA(HIF-1alpha) plus 2 nmol siRNA(PAI-1), resulted in a reduction of abdominal adhesion by 36% and 42%, respectively, with the reduction being statistically significant when compared directly to the saline control (P < 0.01). CONCLUSION: These data show that administration of siRNA/PEI complexes within the peritoneal cavity can be used to prevent post-operative abdominopelvic adhesions.     

Antimicrobial photodynamic therapy combined with conventional endodontic treatment to eliminate root canal biofilm infection

BACKGROUND AND OBJECTIVE: To compare the effectiveness of antimicrobial photodynamic therapy (PDT), standard endodontic treatment and the combined treatment to eliminate bacterial biofilms present in infected root canals. STUDY DESIGN/MATERIALS AND METHODS: Ten single-rooted freshly extracted human teeth were inoculated with stable bioluminescent Gram-negative bacteria, Proteus mirabilis and Pseudomonas aeruginosa to form 3-day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify bacterial burdens. PDT employed a conjugate between polyethylenimine and chlorin(e6) as the photosensitizer (PS) and 660-nm diode laser light delivered into the root canal via a 200-micro fiber, and this was compared and combined with standard endodontic treatment using mechanical debridement and antiseptic irrigation. RESULTS: Endodontic therapy alone reduced bacterial bioluminescence by 90% while PDT alone reduced bioluminescence by 95%. The combination reduced bioluminescence by >98%, and importantly the bacterial regrowth observed 24 hours after treatment was much less for the combination (P<0.0005) than for either single treatment. CONCLUSIONS: Bioluminescence imaging is an efficient way to monitor endodontic therapy. Antimicrobial PDT may have a role to play in optimized endodontic therapy.     

基于PEI的亚氨乙酸型复合螯合吸附材料IAA-PEI-SiO2的制备及其螯合吸附特性

以偶联剂γ氯丙基三甲氧基硅烷为媒介,将聚胺大分子聚乙烯亚胺(PEI)以偶合接枝的方式,接枝于微米级硅胶微粒表面,制得接枝微粒PEISiO2,然后以氯乙酸为试剂,通过亲核取代反应将亚氨乙酸(IAA)基团键合于硅胶微粒表面,形成了具有多齿配基的亚氨乙酸型螯合微粒IAAPEISiO2。重点研究了螯合微粒IAAPEISiO2的制备过程,初步探讨了其对重金属离子及稀土离子的螯合吸附特性。研究结果表明：接枝微粒PEISiO2与氯乙酸之间的亲核取代反应遵循SN2反应历程;反应温度较高或缚酸剂用量过多时,会促进氯乙酸的水解,减缓亲核取代反应,使亚氨乙酸的键合率下降;适宜的反应温度为60 ℃,适宜的缚酸剂NaHCO3用量是使体系中NaHCO3与氯乙酸物质的量之比为1∶1。与接枝微粒PEISiO2相比,螯合微粒IAAPEISiO2对Cu2+及Eu3+的吸附容量大幅度提升,显示出强螯合吸附特性。     

Pulmonary delivery of siRNA against acute lung injury/acute respiratory distress syndrome

The use of small interfering RNAs (siRNAs) has been under investigation for the treatment of several unmet medical needs, including acute lung injury/acute respiratory distress syndrome (ALI/ARDS) wherein siRNA may be implemented to modify the expression of pro-inflammatory cytokines and chemokines at the mRNA level. The properties such as clear anatomy, accessibility, and relatively low enzyme activity make the lung a good target for local siRNA therapy. However, the translation of siRNA is restricted by the inefficient delivery of siRNA therapeutics to the target cells due to the properties of naked siRNA. Thus, this review will focus on the various delivery systems that can be used and the different barriers that need to be surmounted for the development of stable inhalable siRNA formulations for human use before siRNA therapeutics for ALI/ARDS become available in the clinic.     

聚乙烯亚胺介导的报告基因在体外转染效率的研究

目的　探讨阳离子聚合体载体PEI2 5 (分枝状 ,2 5KDa)在体外的转染效率。方法　PEI2 5和LipofectamineTM2 0 0 0作为转染试剂 ,瞬时转染CHO细胞系 ,通过检测荧光素酶来评估PEI和LipofectamineTM2 0 0 0的转染效率。结果　当N P =5 ,6 ,7时 ,PEI2 5的转染效率与LipofectamineTM2 0 0 0相比无显著性差异 (P >0 .0 5 )。结论　经济实用的PEI适用于体外的瞬时转染。     

Phosphorothioated antisense oligodeoxynucleotide suppressing interleukin-10 is a safe and potent vaccine adjuvant

While vaccination is highly effective for the prevention of many infectious diseases, the number of adjuvants licensed for human use is currently very limited. The aim of this study was to evaluate the safety, efficacy, and to clarify the mechanism of a phosphorothioated interleukin (IL)-10-targeted antisense oligonucleotide (ASO) as an immune adjuvant in intradermal vaccination. The cytotoxicity of IL-10 ASO and its ability to promote T cell proliferation were assessed by Cell Counting Kit-8 (CCK-8) assay. The contents of IL-6, IL-8, TNF-α, IL-1β, and IL-10 in inoculated local tissue and the antigen-specific antibody titers in mouse serum samples were determined by ELISA. The target cells of IL-10 ASO were observed using immunofluorescent staining. The results showed that the specific antibody titer of ovalbumin (OVA), a model antigen, was increased 100-fold upon addition of IL-10 ASO as an adjuvant compared to that of OVA alone. IL-10 ASO showed an immunopotentiation efficacy similar to that of Freund’s incomplete adjuvant, with no detectable cell or tissue toxicity. In vitro and in vivo experiments confirmed that IL-10 ASO enhances immune responses by temporarily suppressing IL-10 expression from local dendritic cells and consequently promoting T cell proliferation. In conclusion, IL-10 ASO significantly enhances immune responses against co-delivered vaccine antigens with high efficacy and low toxicity. It has the potential to be developed into a safe and efficient immune adjuvant.