PAZ domain is critical for spermatogenesis in chicken.

Piwi-like protein 1 (PIWIL1) plays a crucial role in stem cell proliferation, embryogenesis, growth, and development. We aimed to unravel the function of PIWIL1 and its Piwi/Argonaute/Zwille (PAZ) domain in chicken embryogenesis. The expression of PIWI1 at different stages of spermatogenesis was analyzed by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and the PAZ domain was mutated based on its 3D structure model using the clustered regularly interspaced short palindromic repeats Cas9 (CRISPR/Cas9) technology. The results indicated that PIWIL1 mRNA was specifically expressed in spermatogonium cells undergoing meiosis. After targeting the PAZ domain (300-370 amino acid residues), we obtained two mutant DF-1 cell clones with 23-bp and 8-bp deletions. Injection of the pCMV-Cas9-puro-sgRNA-2 construct into 2.5-day embryos resulted in generation of 19 different PAZ mutants (13 males and 6 females), which showed delayed hatching, reduced quality of semen, and decreased expression of PIWIL1 and SOX2 at embryonic days 5 and 18. However, we could not obtain PAZ double knockout (KO) chickens by crossing of the F0 generation, suggesting that PAZ double KO may halt embryonic development. Our results indicate that PIWIL1 plays an important role in meiosis and that PAZ mutations can lead to decreased sperm quality, whereas its double KO may arrest embryogenesis in chicken.

Low-temperature thermal reduction of suspended graphene oxide film for electrical sensing of DNA-hybridization.

A reduced graphene oxide (RGO) based capacitive real time bio-sensor was presented. Suspended graphene oxide (GO) film was assembled electrophoretically between the source and drain electrodes of a transistor and then reduced by annealing in hydrogen/nitrogen forming gas to optimize the surface functional groups and conductivity. The resonance frequency of the transmission coefficient (S21) of the transistor was observed to shift with deoxyribonucleic acid (DNA)-hybridization, with a detecting limit of ~5nM. The advantages of the bio-sensing approach include low-noise frequency output, solution based real time detection and capable of on-chip integration.

Recent progress of atomic layer deposition on polymeric materials.

As a very promising surface coating technology, atomic layer deposition (ALD) can be used to modify the surfaces of polymeric materials for improving their functions and expanding their application areas. Polymeric materials vary in surface functional groups (number and type), surface morphology and internal structure, and thus ALD deposition conditions that typically work on a normal solid surface, usually do not work on a polymeric material surface. To date, a large variety of research has been carried out to investigate ALD deposition on various polymeric materials. This paper aims to provide an in-depth review of ALD deposition on polymeric materials and its applications. Through this review, we will provide a better understanding of surface chemistry and reaction mechanism for controlled surface modification of polymeric materials by ALD. The integrated knowledge can aid in devising an improved way in the reaction between reactant precursors and polymer functional groups/polymer backbones, which will in turn open new opportunities in processing ALD materials for better inorganic/organic film integration and potential applications.

Near-infrared upconversion nanoparticles for bio-applications.

Upconversion nanoparticles (UCNs) attract intensive attentions in biomedical applications. They have shown great potential in bioimaging, biomolecule detection, drug delivery, photodynamic therapy and cellular molecules interactions. Due to the anti-Stokes optical property and NIR excitation, UCNs overcome the drawbacks encountered in conventional luminescent biomarkers. High signal to noise ratio, low cytotoxicity and stable high throughput results are obtained using UCNs as luminescent labels or light triggers in biomedical applications. In this review article, the reason for choosing UCNs as biomedical agents, the progress of the UCNs development and case studies of their biomedical applications will be discussed.

Regulation of matrix metalloproteinase-9 protein expression by 1alpha,25-(OH)2D3 during osteoclast differentiation

To investigate 1alpha,25-(OH)2D3 regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1alpha,25-(OH)2D3 during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1alpha,25-(OH)2D3 inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1alpha,25-(OH)2D3 administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.

Alterations of CD4+CXCR5+Tfh cells and its transcription regulatory factors in children with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the alterations of follicular T helper cells (CD4(+)CXCR5(+)Tfh cells, Tfh) on circulating T lymphocytes in children with asthma, and to study the expression of transcription regulatory factors BCL-6 and BLIMP-1 mRNA.</p><p><b>METHODS</b>Sixty-four children with asthma and 25 healthy controls were enrolled in this study. On the basis of the disease, the children with asthma were classified into acute phase group (n=36) and remission phase group (n=28). The flow cytometry was used to detect the proportion of CD4(+)CXCR5(+)Tfh cells on CD4(+)T lymphocytes. Real-time PCR was performed to detect the levels of BCL-6 mRNA and BLIMP-1 mRNA. The double -antibody Sandwich ELISA was used to detect plasma concentrations of total IgE, IL-2, IL-6 and IL-21.</p><p><b>RESULTS</b>The proportion of CD4(+)CXCR5(+)Tfh cells was significantly higher in the acute group than in the control group and the remission group (P<0.05). Transcription levels of BCL-6 mRNA were significantly higher, while the inhibitory factors BLIMP-1 mRNA was significantly lower in the acute group than in the remission group and control group (P<0.05). The plasma concentration of IL-6 in the acute group increased significantly compared with the control group (P<0.05). Plasma concentrations of total IgE and IL-21 increased significantly, in contrast, plasma IL-2 concentration decreased significantly in the acute group, compared with the control group and the remission group (P<0.05). Correlation analysis showed that both IL-21 and IL-6 concentrations were positively correlated with the proportion of CD4(+)CXCR5(+)Tfh cells (r=0.76, r=0.46 respectively; P<0.05), while IL-2 level was negatively correlated with the proportion of Tfh cells (r=-0.68, P<0.05).</p><p><b>CONCLUSIONS</b>The abnormal proportion of CD4(+)CXCR5(+)Tfh cells might be involved in the immunological pathogenesis of acute asthma in children. The increased expression of BCL-6 mRNA and decreased expression of BLIMP-1 mRNA as well as the alterations of plasma total IgE, cytokines IL-2, IL-6 and IL-21 in microenvironment might be account for the increased proportion of CD4(+)CXCR5(+)Tfh cells in children with acute asthma.</p>

Recent advances of paroxysmal kinesigenic dyskinesia / 中华实用儿科临床杂志

Paroxysmal kinesigenic dyskinesia (PKD) is the most frequently described subtype of paroxysmal dyskinesias.The precipitating factor is usually sudden movement or startle.Clinically,PKD cases suffer involuntary movements including unilateral or bilateral chorea,athetosis,dystonia or ballismus,with preserved consciousness.Family history is commonly noted in idiopathic PKD,but sporadic cases are also reported.Familial PKD is inherited in an autosomal dominant fashion with incomplete penetrance.To date,2 loci 16p11-q12 and 16q13-q22 have been mapped to PKD,although a 3rd locus is also suspected.PRRT2,which was located in 16p12.1,was recently identified as causative gene of PKD.However,culprit genes in the other 2 loci remain to be investigated.The potential mechanism of PKD remains largely unclear and the role of mutant PRRT2 in the pathogenesis of PKD is still unknown.In this review,the recent advances of PKD were summarized and hypothesis regarding the mechanisms of PKD was put up,which may make significant contributions to the diagnosis and treatment of PKD.

In vitro cancer cell imaging and therapy using transferrin-conjugated gold nanoparticles.

Gold nanoparticles were conjugated with transferrin molecules for targeting, imaging and therapy of breast cancer cells (Hs578T, ATCC). Results show that, the transferrin-transferrin receptor-mediated cellular uptake of gold nanoparticles is six times of that in the absence of this interaction. As a consequence, the laser power effective for photothermal therapy of the cancer cells was reduced to values of two orders of magnitude lower. To demonstrate the efficiency of the conjugated gold nanoparticles in selectively targeting cancer cells, the cellular uptake of the gold nanoparticles by noncancerous cells (3T3, ATCC) was also investigated. The cellular uptake by the normal cells is only one fourth of that by the cancerous cells indicating that the transferrin-transferrin receptor interaction plays an important role in controlling the cellular uptake of the gold nanoparticles.

A clinical study on psycho-behavior therapy for premature ejaculation / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of psycho-behavior therapy for premature ejaculation (PE).</p><p><b>METHODS</b>A total of 58 PE patients that met the study criteria were treated by psycho-behavior therapy, 2-3 times a week, for a 6-time course. After the treatment, the therapeutic effect was assessed by observation of the changes in the patients' CIPE-5 scores.</p><p><b>RESULTS</b>The rates of cure, effectiveness, ineffectiveness and overall effectiveness were 46.55% (27/58), 32.76% (19/58), 20.69% (12/58) and 79.31%, respectively. The CIPE-5 scores of the patients were elevated from 7.97 +/- 2.30 before the treatment to 22.50 +/- 6.64 after it, and the differences were statistically significant. The psycho-behavior therapy obviously prolonged the ejaculation latency of the patients, increased the sexual satisfaction of both the patients and their spouses, lessened the patients' sexual anxiety and nervousness, and decreased the difficulty in retarding ejaculation, with statistically significant differences from pretreatment.</p><p><b>CONCLUSION</b>Psychobehavior therapy has remarkable therapeutic effect on premature ejaculation.</p>

Establishment of a transgenic cell model for preliminary screening of chemopreventive agents / 生物工程学报

To develop a GFP transgenic cell model under the transcriptional control of TK promoter adjacent to which ARE enhancer was inserted. Synthetic oligonucleotide ARE motif was annealed and purified then inserted into pTK-GFP to construct the vector of pARE-TK-GFP. The TK and ARE-TK fragments were amplified by PCR and cloned into pEGFP-N1 to reconstruct eukaryotic expression vectors of pTK-GFP/Neo and pARE-TK-GFP/Neo. They were transfected into HepG2 cells and clones resistant G418 were isolated. PDTC and Lentinan were used to induce the cell levels of GFP and the fluorescence was measured using a fluorescence plate reader. The results showed that the induced level of GFP is significantly increased and have dose-dependeny in a certain range. This findings indicated that such a cell model offered a potential platform for preliminary screening of all kinds of natural or synthetic chemopreventive agents.