Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice.

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method for isolating DCs primarily involves inducing bone marrow (BM) differentiation into DCs by injecting large doses (>10 ng/mL) of granulocyte-macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4), making the procedure complex and expensive. In this protocol, using all BM cells cultured in 10 ng/mL GM-CSF/IL-4 medium, after 3-4 half-culture exchanges, up to 2.7 x 107 CD11c+ cells (DCs) per mouse (two femurs) were harvested with a purity of 80%-95%. After 10 days in culture, the expression of CD11c, CD80, and MHC II increased, whereas the number of cells decreased. The number of cells peaked after 7 days of culture. Moreover, this method only took 10 min to harvest all bone marrow cells, and a high number of DCs were obtained after 1 week of culture.

Transdermal Photothermal Sterilization and Abscess Elimination Research of BSA-CuS Nanoparticles in vivo.

The treatment of subcutaneous abscess caused by drug-resistant bacteria is facing great difficulties and receiving more attention. In this work, we employed BSA-CuS nanoparticles as a photothermal reagent to apply photothermal therapy (PTT) to combat drug-resistant bacteria in vitro and subcutaneous abscess in vivo. The BSA-CuS nanoparticles were found to be stable and biocompatible without cytotoxicity toward NIH3T3 and 4T1 cells. In vitro experiments showed that three species of drug-resistant pathogens, including Escherichia coli, Staphylococcus aureus, and Candida albicans, could be effectively sterilized under co-incubation with BSA-CuS nanoparticles and then irradiation with 1064 nm NIR laser via tissue penetration. BSA-CuS nanoparticles together with 1064 nm NIR laser irradiation could also effectively diminish subcutaneous abscesses caused by drug-resistant bacteria on mice under PTT and depth PTT without causing any serious side effects and organic damage in vivo.That is OK, thank you!

Long non­coding RNA HHIP­AS1 inhibits lung cancer epithelial­mesenchymal transition and stemness by regulating PCDHGA9.

The aim of the present study was to investigate the effect of hedgehog­interacting protein antisense RNA 1 (HHIP­AS1) on epithelial­mesenchymal transition (EMT) and cellular stemness of human lung cancer cells by regulating the microRNA (miR)­153­3p/PCDHGA9 axis. Reverse transcription­quantitative PCR was used to compare the expression of HHIP­AS1 in lung cancer and adjacent normal lung tissues. In addition, the correlation of HHIP­AS1 with E­cadherin, Vimentin, N­cadherin and Twist1 was analyzed. HHIP­AS1 overexpression vector was transfected into lung cancer A549 and NCI­H1299 cell lines. Cell Counting Kit­8 and Transwell and clonogenic assays were used to detect the proliferation, invasion and clonogenesis of the lung cancer cells, respectively. The associations among HHIP­AS1, miR­153­3p and PCDHGA9 were predicted by bioinformatics analysis and verified by a dual­luciferase reporter system. The results showed that the expression of HHIP­AS1 in lung cancer tissues was significantly lower than that in normal tissues (P<0.001). HHIP­AS1 was positively correlated with E­cadherin and negatively correlated with Vimentin, N­cadherin and Twist1. HHIP­AS1 overexpression inhibited the proliferation, invasion and clonal formation of the A549 and NCI­H1299 cells. The luciferase reporter system verified that HHIP­AS1 could adsorb miR­153­3p and that PCDHGA9 was the target gene of miR­153­3p. A549 cells were transfected with HHIP­AS1 overexpression vector and miR­153­3p mimic, and the miR­153­3p mimic had a mitigating effect on HHIP­AS1 inhibition (P<0.001). In conclusion, HHIP­AS1 inhibits the EMT and stemness of lung cancer cells by regulating the miR­153­3p/PCDHGA9 axis. Thus, HHIP­AS1 may be a new potential target for lung cancer treatment.

A systematic review and meta-analysis of diagnostic performance and physicians' perceptions of artificial intelligence (AI)-assisted CT diagnostic technology for the classification of pulmonary nodules.

BACKGROUND: Lung cancer was the second most commonly diagnosed cancer and the leading cause of cancer death in 2020. Although artificial intelligence (AI)-assisted diagnostic technologies have shown promise and has been used in clinical practice in recent years, no products related to AI-assisted CT diagnostic technologies for the classification of pulmonary nodules have been approved by the National Medical Products Administration in China. The objective of this article was to systematically review the diagnostic performance of AI-assisted CT diagnostic technology for the classification of pulmonary nodules as benign or malignant and to analyze physicians' perceptions of this technology in China. METHODS: All relevant studies from 6 literature databases were searched and screened according to the inclusion and exclusion criteria. Data were extracted and the study quality was assessed by two reviewers. The study heterogeneity and publication bias were estimated. A questionnaire survey on the perceptions of physicians was conducted in 9 public tertiary hospitals in China. A meta-analysis, meta-regression and univariate logistic model were used in the systematic review and to explore the association of physicians' perceptions with their rate of support for the clinical application of the technology. RESULTS: Twenty-seven studies with 5,727 pulmonary nodules were finally included in the meta-analysis. We found that the quality of the included studies was generally acceptable and that the pooled sensitivity and specificity of AI-assisted CT diagnostic technology for the classification of pulmonary nodules as benign or malignant were 0.90 and 0.89, respectively. The pooled diagnostic odds ratio (DOR) was 70.33. The majority of the surveyed physicians in China perceived "reduced workload for radiologists" and "improved diagnostic efficiency" as the important benefits of this technology. In addition, diagnostic accuracy (including misdiagnosis) and practical experience were significantly associated with whether physicians supported its clinical application. CONCLUSIONS: In the context of lung cancer diagnosis, AI-assisted CT diagnostic technology for the classification of pulmonary nodules as benign or malignant has good diagnostic performance, but its specificity needs to be improved.

Cucurbitacin B enhances apoptosis in gefitinib resistant non­small cell lung cancer by modulating the miR­17­5p/STAT3 axis.

Tyrosine kinase inhibitors, such as gefitinib, are currently widely used as targeted therapeutics for non­small cell lung cancer (NSCLC). Although drug resistance has become a major obstacle to successful treatment, mechanisms underlying resistance to gefitinib remain unclear. Therefore, the present study aimed to investigate the impact of adjunctive cucurbitacin B (CuB) on gefitinib resistance (GR) in the PC9 cell line, including identifying underlying mechanisms. Reverse transcription­quantitative PCR demonstrated significant downregulation of microRNA (miR)­17­5p expression in GR PC9 cells (PC9/GR), and this could be reversed by CuB. During combination treatment with CuB and gefitinib at IC25, PC9/GR cell proliferation was downregulated, and apoptosis was upregulated. The presence of a miR­17­5p inhibitor negated the effects of CuB and gefitinib, whereas the presence of a miR­17­5p mimic enhanced them. Luciferase assays demonstrated that the hypothetical target gene, signal transducer and activator of transcription 3 (STAT3), was directly targeted by miR­17­5p. Moreover, significant elevation of the STAT3 protein and phosphorylation levels in PC9/GR cells was reversed by the addition of CuB, despite a lack of change in STAT3 transcription level. During combined treatment with CuB and gefitinib at IC25, the STAT3 protein expression was negatively associated with the expression of miR­17­5p. Overexpression of STAT3 increased proliferation and decreased apoptosis and the protein levels of apoptosis­related factors cleaved caspase­3 and cleaved caspase­9 of PC9/GR cells. Findings indicated that STAT3 protein and phosphorylation levels became elevated in response to gefitinib, and that CuB­induced miR­17­5p expression led to STAT3 degradation, thereby ameliorating GR. In summary, CuB reduced the proliferation of GR PC9 cells by modulating the miR­17­5p/STAT3 axis, and may represent a promising potential novel strategy for the reversal of GR.

The potential diagnosis role of TP53 mutation in advanced bladder cancer: A meta-analysis.

BACKGROUND: Bladder cancer is one of the most common urological cancers all over the world, and NMIBC occupies almost 80% of recently diagnosed bladder cancer cases. Progress and recurrence of bladder cancer are the main problems during the disease. The level of TP53 mutation is obviously higher in the high stage than the lower. This meta-analysis is to evaluate the potential diagnosis feature of TP53 mutation by the expression of TP53 mutation of Ta stage vs high stage in bladder cancer. METHODS: A systematic search of databases was conducted, and some relevant articles were selected. Next, the meta-analysis was carried out according to the standard guidelines. RESULTS: There were seven researches in which 677 participants were selected at the basis of inclusion standard. TP53 mutation was associated highly with increased diagnosis of bladder cancer. We found that the high stage of bladder cancer has obviously higher level of TP53 mutation than the lower stage, and these patients of MIBC have higher expression of TP53 mutation compared with NMIBC. No significant publication bias has been observed in this meta-analysis. The expression of TP53 mutation might be a diagnose-related biomarker for lots of patients with bladder cancer. CONCLUSIONS: The results of this meta-analysis provided further evidences that the expression of TP53 mutation was associated with the diagnosis efficiency of advanced bladder cancer. Higher expression of TP53 mutation was observed in the high stage of bladder cancer or the MIBC, and lower expression of TP53 mutation in the Ta stage of bladder cancer or the NMIBC. The expression level of TP53 mutation was probably a critical diagnosed biomarker in advanced bladder cancer.

Well-water-dispersed N-trimethyl chitosan/Fe3O4 hybrid nanoparticles as peroxidase mimetics for quick and effective elimination of bacteria.

Fe3O4 nanoparticles, used as peroxidase mimetics, exhibit splendid future in the biomedical field. However, the functionalization on Fe3O4 nanoparticles always goes with the loss of superparamagnetism and decrease in peroxidase-activity. Here, we synthesized green polyethylene glycol (PEG)-functionalized magnetic/N-trimethyl chitosan (CS) hybrid nanoparticles (Fe3O4@PAA/TMC/PEG NPs) with improved water dispersibility, superparamagnetism, high saturation magnetization and well peroxidase-like activity. The functionalized coating was divided in two steps, one involved a cross-linked PEG/PAA/CS middle layer to protect the nanocrystal Fe3O4 from oxidization, the other was a hydrophilic PEG/TMC outer layer improving the water dispersion, biocompatibility, as well as supplying positive quaternary ammonium groups for a potential increase of cell binding efficiency. The structure, composition and morphology of Fe3O4@PAA/TMC/PEG NPs were characterized by TEM, FT-IR spectroscopy, DLS, zeta potential measurement, respectively. Thermal performance was characterized by TGA, and the peroxidase-like mimics activity was tested by TMB·2HCl colour development experiments. The magnetic property of the as-prepared hybrid nanoparticles was first confirmed by VSM, and then proved by the bacterial pathogens adsorption, especially at ultralow pathogen concentration. Particularly, with an external magnet, the Fe3O4@PAA/TMC/PEG NPs, combined cationic quaternary ammonium groups and peroxidise-mimetic catalytic activity, were tested for antibacterial effect by plating method.

Relationship between physician financial incentives and clinical pathway compliance: a cross-sectional study of 18 public hospitals in China.

OBJECTIVES: Many strategies have been either used or recommended to promote physician compliance with clinical practice guidelines and clinical pathways (CPs). This study examines the relationship between hospitals' use of financial incentives to encourage physician compliance with CPs and physician adherence to CPs. DESIGN: A retrospectively cross-sectional study of the relationship between the extent to which patient care was consistent with CPs and hospital's use of financial incentives to influence CP compliance. SETTING: Eighteen public hospitals in three provinces in China. PARTICIPANTS: Stratified sample of 2521 patients discharged between 3 January 2013 and 31 December 2014. PRIMARY OUTCOME MEASURES: The proportion of key performance indicators (KPIs) met for patients with (1) community-acquired pneumonia (pneumonia), (2) acute myocardial infarction (AMI), (3) acute left ventricular failure (heart failure), (4) planned caesarean section (C-section) and (5) gallstones associated with acute cholecystitis and associated cholecystectomy (cholecystectomy). RESULTS: The average implementation rate of CPs for five conditions (pneumonia, AMI, heart failure, C-section and cholecystectomy) based on 2521 cases in 18 surveyed hospitals was 57% (ranging from 44% to 67%), and the overall average compliance rate for the KPIs for the five conditions was 69.48% (ranging from 65.07% to 77.36%). Implementation of CPs was associated with greater compliance within hospitals only when hospitals adopted financial incentives directed at physicians to promote compliance. CONCLUSION: CPs are viewed as important strategies to improve medical care in China, but they have not been widely implemented or adhered to in Chinese public hospitals. In addition to supportive resources, education/training and better administration in general, hospitals should provide financial incentives to encourage physicians to adhere to CPs.

Patient safety climate in general public hospitals in China: differences associated with department and job type based on a cross-sectional survey.

OBJECTIVE: This study analysed differences in the perceived patient safety climate among different working departments and job types in public general hospitals in China. DESIGN: Cross-sectional survey. SETTING: Eighteen tertiary hospitals and 36 secondary hospitals from 10 areas in Shanghai, Hubei Province and Gansu Province, China. PARTICIPANTS: Overall, 4753 staff, including physicians, nurses, medical technicians and managers, were recruited from March to June 2015. MAIN OUTCOME MEASURE: The Patient Safety Climate in Healthcare Organisations (PSCHO) tool and the percentages of 'problematic responses' (PPRs) were used as outcome measures. Multivariable two-level random intercept models were applied in the analysis. RESULTS: A total of 4121 valid questionnaires were collected. Perceptions regarding the patient safety climate varied among departments and job types. Physicians responded with relatively more negative evaluations of 'organisational resources for safety', 'unit recognition and support for safety efforts', 'psychological safety', 'problem responsiveness' and overall safety climate. Paediatrics departments, intensive care units, emergency departments and clinical auxiliary departments require more attention. The PPRs for 'fear of blame and punishment' were universally significantly high, and the PPRs for 'fear of shame' and 'provision of safe care' were remarkably high, especially in some departments. Departmental differences across all dimensions and the overall safety climate primarily depended on job type. CONCLUSIONS: The differences suggest that strategies and measures for improving the patient safety climate should be tailored by working department and job type.

Propagation of multi-cosine-Laguerre-Gaussian correlated Schell-model beams in free space and atmospheric turbulence.

We introduce a class of random stationary, scalar source named as multi-cosine-Laguerre-Gaussian-correlated Schell-model (McLGCSM) source whose spectral degree of coherence (SDOC) is a combination of the Laguerre-Gaussian correlated Schell-model (LGCSM) and multi-cosine-Gaussian correlated Schell-model (McGCSM) sources. The analytical expressions for the spectral density function and the propagation factor of a McLGCSM beam propagating in turbulent atmosphere are derived. The statistical properties, such as the spectral intensity and the propagation factor, of a McLGCSM beam are illustrated numerically. It is shown that a McLGCSM beam exhibits a robust ring-shaped beam array with adjustable number and positions in the far field by directly modulating the spatial structure of its SDOC in the source plane. Moreover, we provide a detailed insight into the theoretical origin and characteristics of such a ring-shaped beam array. It is demonstrated that these peculiar shaping properties are the concentrated manifestation of the individual merits respectively associated with the Laguerre- and multi-cosine-related factors of the whole SDOC. Our results provide a novel scheme to generate robust and controllable ring-shaped beam arrays over large distances, and will widen the potentials for manipulation of multiple particles, free-space optical communications and imaging in the atmosphere.

[EFFECT OF α-LIPOIC ACID IN INHIBITING OXIDATIVE STRESS AND PROMOTING DIABETIC WOUND HEALING BY SUPPRESSING EXPRESSION OF miR-29b IN MICE].

OBJECTIVE: To investigate the effect of α-lipoic acid on the oxidative stress of wound tissues and diabetic wound healing in mice with diabetic feet. METHODS: Sixty male C57BL/6J mice weighting 200-300 g were randomly divided into model group (control group, n=15), α-lipoic acid-treated model group (n=15), miR-29b mimic group (n=15), and miR-29b mimic negative control group (NC group, n=15). All animals received intraperitoneal injection of streptozocin to establish the diabetic model. Then, a full thickness wound of 5 mm×2 mm in size was created at 4 weeks after modeling. All mice were administrated with high-sugar-fat-diet. At the same day after modeling, α-lipoic acid-treated model group was continuously given intravenous injection of 100 mg/(kg·d) α-lipoic acid for 14 days; miR-29b mimic group and NC group received the tail intravenous injection of lentiviral vector for miR-29b mimic and miR-29b mimic negative control (a total of 2×107 TU), respectively, with the treatment of α-lipoic acid. The wound healing was observed and wound area was measured at 7 and 14 days. The wound tissues were harvested to detect the levels of superoxide dismutase (SOD) and glutathione (GSH) using xanthine oxidase method and 5, 5-dithiobis-2-nitrobenzoic acid staining method at 14 days. At the same day, 7, and 14 days after modeling, the relative miR-29b expression in wound tissues from control and α-lipoic acid-treated model groups was detected by real-time fluorescence quantitative PCR. RESULTS: All mice survived to the experiment end. The wound healing was faster in α-lipoic acid-treated group than control group. At 7 and 14 days, the relative wound area and miR-29b expression level were significantly lower, while the contents of SOD and GSH were significantly higher in α-lipoic acid-treated group than control group (P<0.05). In addition, miR-29b mimic group had significantly increased relative wound area and significantly decreased the contents of SOD and GSH when compared with NC group at 7 and 14 days (P<0.05). CONCLUSIONS: α-lipoic acid could inhibit oxidative stress and promote diabetic wound healing by suppressing expression of miR-29b in mice.

In vivo implantation of hydrophobic acrylic intraocular lenses with surface modification.

PURPOSE: To assess the surface properties of modified hydrophobic acrylic intraocular lenses (IOL) implanted in rabbits. METHODS: The hydrophobic acrylic IOLs were modified with monomer vinyl pyrrolidone by surface modification technique. Phacoemulsification combined with IOL implantation was conducted in 9 rabbits (18 eyes). Postoperative responses were observed by slit-lamp microscope at 3, 7, 15, 30, 90 days after surgery. RESULTS: During the early stage after IOL implantation, corneal edema and anterior chamber fibrin exudation were observed. The exudate fluid was almost absorbed at the 15th day postoperatively. At 7th day, the anterior chamber exudation in the modification group was significantly less severe than that in non-modification group (P < 0.05). Posterior capsular opacification occurred at 30th day after surgery and was aggravated 90 days later. IOL dislocation was seen in 5 eyes and occlusion of pupil in 3 eyes. CONCLUSION: The hydrophobic acrylic IOLs with surface modification have improved surface properties and higher uveal biocompatibility.

Study on the propagation parameters of Bessel-Gaussian beams carrying optical vortices through atmospheric turbulence.

Based on the integral representation of Bessel function and the extended Huygens-Fresnel principle, an integral expression of the Wigner distribution function (WDF) for partially coherent Bessel-Gaussian beams (PBGBs) propagating through turbulent atmosphere has been obtained. Also, the analytical formulas of the M2-factor for PBGB propagation in such a medium have been derived, which can be applied to cases of different spatial power spectra of the refractive index fluctuations. The performed numerical results reveal that the M2-factor of a PBGB in turbulent atmosphere depends on the beam parameters of the initial input beam, the structure constants of the turbulent atmosphere, and the propagation distance. These results may be useful in long-distance optical communications in free space or in turbulent atmosphere.

Changes in growth hormone (GH), GH receptor, and GH signal transduction in hippocampus of congenital hypothyroid rats.

Recent studies have shown that, like thyroid hormone (TH), growth hormone (GH) plays a critical role in development of the brain. However, it is still unclear whether the functions of the two hormones are locally orchestrated in the brain or whether TH has a permissive effect on GH in the central nervous system as it does in the periphery. To address this question, the present study investigated the changes in local expression of GH and GH receptor (GHR) and the activity of GH signaling molecules in the hippocampus of congenitally hypothyroid (CHT) rats. As demonstrated by morphometric measurements and the Y-maze test, CHT rats had decreased neurons and weaker Nissl staining in the stratum pyramidal/granule in the hippocampus and a reduced acquisition of safe place recognition memory. Analyses of QPCR and Western blot revealed a substantially decreased hippocampal expression of GH and GHR, accompanied by a corresponding decrease in phosphorylation of JAK2 and STAT5 in the CHT rats. These changes were, at least in part, corrected by systemic supplement of T3. The findings provide the first direct evidence suggesting that the functional autocrine and paracrine regulation of GH in the CNS is orchestrated by TH.

Upregulation of Tanis mRNA expression in the liver is associated with insulin resistance in rats.

Tanis, a newly discovered membrane protein, has been suggested to be involved in the development of insulin resistance (IR). In addition, Tanis was postulated as a hepatic receptor for an inflammatory marker, serum amyloid A-1 (SAA1), which represents a risk factor for cardiovascular disease. The aim of this study was to investigate the potential role of Tanis and SAA1 in the development of IR using a rat model. Male Sprague-Dawley (6 weeks old) rats were divided into three groups (n = 10 for each group): control, IR, and rosiglitazone-treated IR. Rosiglitazone is an insulin sensitizer that is used for treatment of type 2 diabetes. Rats were fed a high-fat diet for 35 days to induce IR, as judged by increased plasma glucose and insulin concentrations, following an oral glucose tolerance test. Rats of three groups were sacrificed, and the livers were dissected for RNA preparation. Real-time PCR analysis revealed that Tanis and SAA1 mRNA levels were significantly higher in the livers of IR rats than those of control rats fed a standard diet and those of IR rats treated for 35 days with rosiglitazone. Importantly, there was no significant difference in Tanis and SAA1 mRNA levels between control and rosiglitazone-treated IR rats. Furthermore, the expression levels of Tanis mRNA are positively correlated with those of SAA1 mRNA in the livers of control, IR and rosiglitazone-treated IR rats. In conclusion, high expression of Tanis mRNA in the liver is associated with the development of IR probably via the function of SAA1.

Virtual sources for coherent combination beams consisting of off-axis Gaussian beams.

Based on the operator transformation technique, the multiple complex point sources required to generate a coherent superposition of waves are introduced and a closed-form analytical expression is derived for this composite wave. From the expression of the composite wave, the paraxial approximation and the nonparaxial corrections of all orders for the corresponding paraxial beam are determined. The paraxial composite beam uniformly represents off-axis Gaussian beams (GBs), sin(cos)-GBs, sinh(cosh)-GBs, nth-order modified Bessel-GBs, and Bessel-GBs with topological charge.

Variation in gene expression of presenilins-associated rhomboid-like protein and mitochondrial function in skeletal muscle of insulin-resistant rats.

Multiple factors promote insulin resistance. In this study, we evaluated the mRNA levels of presenilins-associated rhomboid-like protein (PARL) and mitochondrial content and enzyme activity from skeletal muscle isolated from insulin-resistant rats. Rats fed a high-fat diet for 35 days developed moderate insulin resistance, which was determined by an increase in plasma glucose and insulin concentrations following an oral glucose tolerance test. The PARL mRNA level was lower in the insulin-resistant rats than in control animals, and is associated with low mitochondrial content and reduced mitochondrial enzyme activity in the skeletal muscle from the insulin-resistant rats. The results suggest that high-fat-diet-induced insulin resistance is associated with mitochondrial dysfunction in skeletal muscle, and may be the result of the decreased expression of the PARL gene, which encodes the protein with functional significance in mitochondria.

Propagation of Bessel-Gaussian beams with optical vortices in turbulent atmosphere.

An approximate expression of a Bessel-Gaussian beam (BGB) with desired topological charge is introduced using a coherence superposition of decentered Gaussian beams (dGBs). And based on such an expression and the extended Huygens-Fresnel principle, the propagation properties of BGBs traveling in turbulent atmosphere are explored. An analytical expression of the average intensity of a BGB with phase singularity propagating through turbulent atmosphere is obtained and analyzed numerically. It is found that intensity profiles of BGBs experienced successive variations and the phase singularity rapidly fades away during propagating in turbulent atmosphere.

Expression of human insulin gene wrapped with chitosan nanoparticles in NIH3T3 cells and diabetic rats.

AIM: To study the expression of human insulin gene wrapped with chitosan nanoparticles in NIH3T3 cells and diabetic rats. METHODS: pCMV.Ins, an expression plasmid of the human insulin gene, was constructed. In total, 100 microg pCMV.Ins wrapped with chitosan nanoparticles (chitosan-pCMV.Ins) was transfected to NIH3T3 cells and diabetes rats through lavage and coloclysis, respectively. The transfected cells were grown in Dulbecco's modified Eagle's medium, containing G418, for 72 h after transfection. The clones were selected and continued to grow in G418 medium for 24 d. The expression of human insulin was detected by immunohistochemistry. Human insulin in the culture medium of transfected cells was measured. Fasting blood glucose and plasma human insulin of diabetic rats were measured for 5 d after transfection. RT-PCR and Western blotting were performed to confirm the expression of the human insulin gene in diabetic rats. RESULTS: Approximately 10% of NIH3T3 cells transfected by chitosan-pCMV.Ins expressed human insulin. Human insulin in the culture medium of NIH3T3 cells transfected by chitosan-pCMV.Ins significantly increased compared with that of the control group (P<0.01). Fasting blood glucose levels of the lavage group and the coloclysis group decreased significantly in 5 d (P<0.01) in comparison, while plasma insulin levels were much higher (P<0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and the coloclysis groups. CONCLUSION: The human insulin gene can be transfected and expressed successfully by chitosan- pCMV.Ins in NIH3T3 cells and diabetes rats, which indicates that chitosan is a promising, non-viral vector for gene expression.

Vortex characteristics of Fraunhofer diffractions of a plane wave by a spiral phase plate limited by pseudoring polygonal apertures.

We introduce a multilevel spiral phase plate (SPP) limited by a pseudoring polygonal aperture (PRPA). Such an SPP has the advantages of easier fabrication and greater suppression of the sidelobes of the diffraction field over that generated with a polygonal aperture (PA). The Fraunhofer diffraction fields generated by an SPP with a PRPA or with a PA have the same topological charge features and a similar diffraction pattern. Numerical evaluations show that the maximum bright annular-intensity difference between the diffraction patterns for the SPP with a PRPA and that of a PA does not exceed 3% under optimal design parameters.