Fabrication of three-dimensional porous scaffolds with controlled filament orientation and large pore size via an improved E-jetting technique.

Biodegradable polymeric scaffolds have been widely used in tissue engineering as a platform for cell proliferation and subsequent tissue regeneration. Conventional microextrusion methods for three-dimensional (3D) scaffold fabrication were limited by their low resolution. Electrospinning, a form of electrohydrodynamic (EHD) printing, is an attractive method due to its capability of fabricating high-resolution scaffolds at the nanometer/micrometer scale level. However, the scaffold was composed of randomly orientated filaments which could not guide the cells in a specific direction. Furthermore, the pores of the electrospun scaffold were small, thus preventing cell infiltration. In this study, an alternative EHD jet printing (E-jetting) technique has been developed and employed to fabricate 3D polycaprolactone (PCL) scaffolds with desired filament orientation and pore size. The effect of PCL solution concentration was evaluated. Results showed that solidified filaments were achieved at concentration >70% (w/v). Uniform filaments of diameter 20 µm were produced via the E-jetting technique, and X-ray diffraction and attenuated total reflectance Fourier transform infrared spectroscopic analyses revealed that there was no physicochemical changes toward PCL. Scaffold with a pore size of 450 µm and porosity level of 92%, was achieved. A preliminary in vitro study illustrated that live chondrocytes were attaching on the outer and inner surfaces of collagen-coated E-jetted PCL scaffolds. E-jetted scaffolds increased chondrocytes extracellular matrix secretion, and newly formed matrices from chondrocytes contributed significantly to the mechanical strength of the scaffolds. All these results suggested that E-jetting is an alternative scaffold fabrication technique, which has the capability to construct 3D scaffolds with aligned filaments and large pore sizes for tissue engineering applications.

Electroacupuncture Could Regulate the NF-κB Signaling Pathway to Ameliorate the Inflammatory Injury in Focal Cerebral Ischemia/Reperfusion Model Rats.

The activated nuclear factor-KappaB signaling pathway plays a critical role in inducing inflammatory injury. It has been reported that electroacupuncture could be an effective anti-inflammatory treatment. We aimed to explore the complex mechanism by which EA inhibits the activation of the NF- κ B signal pathway and ameliorate inflammatory injury in the short term; the effects of NEMO Binding Domain peptide for this purpose were compared. Focal cerebral I/R was induced by middle cerebral artery occlusion for 2 hrs. Total 380 male Sprague-Dawley rats are in the study. The neurobehavioral scores, infarction volumes, and the levels of IL-1 ß and IL-13 were detected. NF- κ B p65, I κ B α , IKK α , and IKK ß were analyzed and the ability of NF- κ B binding DNA was investigated. The EA treatment and the NBD peptide treatment both reduced infarct size, improved neurological scores, and regulated the levels of IL-1 ß and IL-13. The treatment reduced the expression of IKK α and IKK ß and altered the expression of NF- κ B p65 and I κ B α in the cytoplasm and nucleus; the activity of NF- κ B was effectively reduced. We conclude that EA treatment might interfere with the process of NF- κ B nuclear translocation. And it also could suppress the activity of NF- κ B signaling pathway to ameliorate the inflammatory injury after focal cerebral ischemia/reperfusion.

[Cloning of AGO2 and Dcr-2 gene fragments and analysis of their transcription level in different developmental stages of Aedes albopictus].

OBJECTIVE: To perform molecular cloning of the AGO2 and Dcr-2 gene fragments associated with RNA interference pathway of Aedes albopictus and characterize the transcription level of the two genes across all life stages of the mosquito. METHODS: The degenerate primers were designed based on the conserved regions of AGO2 and Dcr-2 amino acid sequences, and the AGO2 and Dcr-2 cDNA fragments were amplified from total RNA of a female mosquito by RT-PCR. The PCR products were cloned into pMD18-T vector and transformed into E. coli DH5alpha strain, and the positive clones were selected and sequenced, with the results for homology analysis by Blastx. The specific primers were designed according to the sequences of AGO2 and Dcr-2 from Ae. albopictus, which were used to investigate the transcription levels of these two genes from eggs, I and II instars larvae, III and IV instars larvae, pupa, male and female mosquitoes by semi-quantitative RT-PCR. RESULTS: The AGO2 and Dcr-2 cDNA fragments obtained were 326 bp and 491 bp in length, with the Accession number of JQ764670 and JQ764671, respectively. The Blastx analysis showed that the AGO2 and Dcr-2 amino acid sequences shared 91% similarity to AGO2 of Ae. aegypti and 98% to Dcr-2 of Ae. albopictus. The transcription of AGO2 and Dcr-2 genes was detected in all life stages of Ae. albopictus, with the highest level of mRNA in female mosquitoes, which was 3.1 times and 15.5 times higher for AGO2 and Dcr-2 than in male mosquitoes, respectively, and significantly higher than other developmental stages (P<0.05). CONCLUSION: The AGO2 and Dcr-2 cDNA sequences have been partially obtained and the highest transcription level found in female Ae. albopictus, suggesting that AGO2 and Dcr-2 are the key genes of RNA interference in female mosquitoes.

[Effects of electroacupuncture on endothelial progenitor cells in peripheral blood and bone marrow in local cerebral ischemia/reperfusion rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) of "Hegu" (LI 4) on blood stromal cell-derived factor 1alpha (SDF-1alpha), CXC chemokine receptor 4+ (CXCR 4+) positive cells and endothelial progenitor cells (EPCs) and bone marrow (BM) EPCs levels in local cerebral ischemia/reperfusion injury (CI/IR) rats, so as to study its mechanisms underlying improvement of cerebral ischemia. METHODS: A total of 54 SD rats were randomly divided into normal control, model, and EA groups. The latter two groups were further divided into 1 d, 2 d, 3 d and 7 d subgroups (4 time-points), respectively. CI/RI model was established by occlusion of the middle cerebral artery. EA (40 Hz/60 Hz, 1-2 mA) was applied to bilateral "Hegu" (LI 4) for 15 min, once daily, 1, 2, 3 and 7 days, respectively. Flow cytometer was used to detect the counts of EPCs and CXCR 4+ cells in the peripheral blood, and BM EPCs. Serum SDF-1alpha was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with the normal control group, the percentages of blood EPCs and CXCR 4+ cells and BM EPCs, and serum SDF-1alpha content on the 1st day, and the percentage of blood CXCR 4+ cells on the 2nd day were increased significantly in the model group (P<0.01, P<0.05). In comparison with the model group, percentages of blood EPCs and CXCR 4+ cells on the 1st day of EA group were reduced significantly (P<0.01), but the percentage of blood EPCs on the 2nd day, those of blood CXCR 4+ cells on the 2nd day and 3rd day, serum SDF-1alpha content on the 2nd day, and the percentage of BM EPCs on the 2nd day in the EA group were all up-regulated significantly (P<0.05, P<0.01). No significant differences were found among the 3 groups in blood EPCs percentages on the 3rd day and 7th day, in blood SDF-1alpha content and BM EPCs percentage on the 7th day (P>0.05). CONCLUSION: Generally, 2 days' EA of LI 4 can up-regulate the percentages of blood and BM EPCs and blood CXCR 4+ cells as well as blood SDF-1alpha protein content in CI/RI rats, which may contribute to its effect in improving cerebral ischemia.

Digit length ratio and male fertility among infertile patients in Ningxia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association between the digit length ratio (DLR) and male fertility in Ningxia.</p><p><b>METHODS</b>Using an electronic sliding caliper, we measured the lengths of the index finger (D2), middle finger (D3), ring finger (D4) and little finger (D5) of both the right and left hand of 136 infertile males in Ningxia, 45 of them with normal and the other 91 with abnormal semen, including 28 cases of oligozoospermia, 10 cases of obstructive azoospermia (OA) and 53 cases of nonobstructive azoospermia (NOA). We calculated the DLRs of D2:D3, D2:D4, D2:D5, D3:D4, D3:D5 and D4:D5 and compared them among different groups.</p><p><b>RESULTS</b>The mean values of FLR presented a pattern of D2:D3 < D2:D4 < D3:D4 < D2:D5 < D4:D5 in the infertile males. DLRs of D2:D3, D2:D4, D2:D5, D3:D5 and D4:D5 of the right hand were significantly higher in the abnormal than in the normal semen group (t = 2.56, 2.48, 3.57, 2.85, 2.53; P < 0.05); those of D2:D3, D2:D4, D2:D5 and D4:D5 of the right hand presented a pattern of NOA > oligozoospermia > OA > normal semen group, while that of D3:D5 presented a pattern of NOA > OA > oligozoospermia > normal semen group. The NOA group showed a significantly higher DLR of the right hand than the normal semen group in D2:D3, D2:D4, D3:D5 and D4:D5, the oligozoospermia group in D2:D4, D3:D5 and D4:D5, and the OA group in D2:D3 (P < 0.05). The D4:D5 DLR of the right hand was remarkably higher in the Han than in the Hui ethnic group (t = 2.01, P < 0.05).</p><p><b>CONCLUSION</b>Higher DLRs of D2:D3, D2:D4, D2:D5, D3:D5 and D4:D5 of the right hand were associated with lower fertility in infertile males in Ningxia. And the D4:D5 FLRs of the right hand may be different between Hui and Han infertile men in Ningxia.</p>