Progress of researches on serine protease inhibitors for zoonotic cestode family Taeniidae / 中国血吸虫病防治杂志

Serine protease inhibitor, a protein superfamily that inhibits the serine protease activity, protects hosts from parasitic infections. This review describes the spatial structure and classification of serine protease inhibitor, mechanisms underlying the interplay between serine protease inhibitor and host immune responses and current advances in serine protease inhibitor of zoonotic cestode family Taeniidae, so as to provide insights into the diagnosis of zoonotic tapeworm infections, discovery of therapeutic targets and screening of vaccine candidates.

Advances in Research of Drugs for Treating Respiratory Syncytial Virus / 中国实验方剂学杂志

Respiratory syncytial virus (RSV) is an RNA virus, which belongs to the paramyxoviridae family, and is transmitted by air droplets and close contact and the main pathogen causing acute lower respiratory tract infection in infants, the elderly and immunocompromised individuals.Although there have been studies on the prevention and treatment of RSV drugs and RSV infection in patients, many medical demands have not been met.And there is no specific antiviral therapy.The only two drugs approved to be applied in RSV prevention and treatment are perizol and ribavirin.However, the former must be used prophylactically, and primarily in high-risk children, while the latter is less effective, and some children even suffer from airway spasm.Therefore, it is urgent to propose new methods for prevention and treatment of RSV.In recent years, traditional Chinese medicine(TCM) has shown a good anti-RSV effect, with a fewer side effects, less resistance to drugs and broad-spectrum antiviral advantage.There are also newly developed biological and chemical anti-RSV drugs.Some new drugs have shown a good efficacy, with an extended half-life and reduced costs, such as fusion inhibitors, monoclonal antibodies.This paper reviews the research progress of anti-RSV drugs in the aspects of TCM, biological drug and chemical drugs, laying a foundation for the development of new anti-RSV drugs and the formulation of new therapeutic strategies.

Meta-analysis of internal fixation versus hip replacement in the treatment of trochanteric fractures / 中国骨伤

<p><b>OBJECTIVE</b>To compare the efficacy of internal fixation (including PFNA and PFN) versus hip replacement (including FHR or THA) in the treatment of trochanteric fractures in adults.</p><p><b>METHODS</b>Reports of studies using randomized controlled trials (RCT) to compare internal fixationg with hip replacement in the management of intertrochanteric fractures were retrieved (up to January 1, 2013) from the Cochrane Library, PUBMED Data, CNKI (China National Knowledge infrastructure), Elsevier, the Chinese Biomedical Database, Wanfang Data, and manually. Methodological quality of the trials was critically assessed, and relevant data were extracted. Statistical software RevMan 5.0 was used for data-analysis.</p><p><b>RESULTS</b>Seven articles were included in the meta-analysis. The results showed that,compared internal fixation with hip replacement,there were statistical significance in the duration of surgery time [WMD = -2.66, 95% CI (-5.25,-0.06), P = 0.05], intra-operative blood loss [WMD = -24.20, 95% CI (-30.38, -18.02), P < 0.000 01], hospital stays time [WMD = -4.72, 95% CI (-5.18, -4.25), P < 0.000 01], bearing load time [WMD = -29.54, 95% CI (-30.77, -28.31), P < 0.000 01], total complications rate [WMD = 0.15, 95% CI (0.11, 0.22), P < 0.000 01], the good rate of Harris scores [WMD = 1.09, 95% CI (0.54,1.32), P < 0.05]. However, there were no statistical significance in the rate of deep venous thrombosis [WMD = 1.09, 95% CI (0.47, 2.55), P > 0.05]. CON- CLUSION: Hip replacement (containing FHR or THA) for the treatment of intertrochanteric fractures is superior to internal fixa- tion in regards to the duration of surgery time, the mean duration of hosipital stays, mean post-operative down time, intra-opera- tive blood loss, the rate of post-operative good Harris scores. But there is not enough evidence to show any difference between hip replacement (containing THA or FHR) and internal fixation in regards to the rate of deep venous thrombosis. However, internal fixation for the treatment of intertrochanteric fractures is superior to hip replacement (containing FHR or THA) in regards to total complications rate.</p>

Effect of bone marrow mesenchymal stem cells on tumor neovascularization / 中华病理学杂志

<p><b>OBJECTIVE</b>The effect of human bone marrow mesenchymal stem cells (hMSCs) on tumor neovascularization were studied.</p><p><b>METHODS</b>hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. hMSCs-EGFP were obtained by pLEGFP-N1 retroviral vector. Flow cytometry was used to detect the cell surface antigen and the differentiation potential of hMSCs-EGFP was investigated under conditioned media. The effect of hMSCs on tumor neovascularization were observed by establishing solid tumor models in BALB/C nude mice. In addition, effect of the conditioned medium used for tumor cells and endothelial cells (EC) cultivation was collected, to detect its effect on the growth and migration rates of hMSC. hMSCs were induced to differentiate into EC in vitro and the migratory effect on HUVEC was also evaluated.</p><p><b>RESULTS</b>hMSCs-EGFP, like hMSC, exhibited a fibroblast-like morphological feature, and both had the similar cell surface antigens. They could be induced into osteocytes or adipocytes under the conditioned media. The results not only suggested that hMSCs contributed to tumor neovascularization, but also indicated that most of vessels were host-derived angiogenesis mediated by hMSCs. The mean vascular density (MVD) in suspension group (13.67 ± 1.53) was strikely higher than that in MCF-7 group (5.33 ± 1.42), which showed statistical significance (P < 0.05). Only very few vessels were attributed to hMSCs transdifferentiation into ECs. Tumor cells and ECs can promote hMSCs proliferation and migration through paracrine action. Furthermore, hMSCs were positive for CD31 after 2 weeks induction and HUVEC migration can be facilitated by hMSCs.</p><p><b>CONCLUSION</b>MSCs have the effect of promoting tumor neovascularization.</p>

Role of aquaporin 1 in the migration of eosinophils from asthmatic guinea pigs / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the role of aquaporin 1 (AQP1) in the migration of eosinophils (EOS) and to determine if AQP-1 can be viewed as the chemotactic activity marker of EOS.</p><p><b>METHODS</b>Asthma model of guinea pigs were developed and EOS were purified from both peripheral blood and bronchoalveolar lavage fluid (BALF). The smears of EOS were studied by in situ hybridization for determining AQP1 mRNA and immunofluorescence under laser scanning confocal microscope for determining AQP1 protein.</p><p><b>RESULTS</b>AQP1 was found expressed in EOS both from peripheral blood and from BALF. Compared with the expression of AQP1 mRNA (mean grey value 109.200 +/- 5.756, x +/- s) and protein (average fluorescence intensity 279.926 +/- 11.293) in EOS from BALF, there was stronger expression of AQP1 mRNA (92.904 +/- 3.290) and protein (425.081 +/- 17.474) in EOS from peripheral blood. The difference both of AQP1 mRNA (t = 9.519, P < 0.05) and protein(t = 27.020, P < 0.05) were considered statistically significant respectively.</p><p><b>CONCLUSIONS</b>AQP1 plays a crucial role in EOS movement. It is possible that EOS produce more AQP1 protein to accelerate its migration to inflammatory tissues under allergic disease and EOS with AQP1 highly expressed are activated. AQP1 can be viewed as the chemotactic activity marker of EOS.</p>

Mesenchymal stem cells transduced by PLEGFP-N1 retroviral vector maintain their biological features and differentiation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Enhanced green fluorescent protein (EGFP) has been an important reporter gene for gene therapy. Human mesenchymal stem cells (hMSCs) are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction.</p><p><b>METHODS</b>hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector, and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated.</p><p><b>RESULTS</b>The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs.</p><p><b>CONCLUSIONS</b>hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.</p>

Effect of wortmannin on endothelial cell proliferation and migration induced by high glucose Müller cell conditioned medium / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of wortmannin on endothelial cells proliferation and migration induced by high glucose Müller cell conditioned medium (HGMCM).</p><p><b>METHODS</b>Immunofluorescence, flow cytometry and Boyden chamber migration models were used to analyze the effect of wortmannin on endothelial cells.</p><p><b>RESULTS</b>50 nmol/L wortmannin could significantly inhibit the proliferation and migration of endothelial cells induced by HGMCM. The percentage of endothelial cells in S phase was obviously decreased [from (37.82 +/- 0.57)% to (21.91 +/- 0.23)%, P < 0.01], while there was an increase in the percentage of cells in G(0)/G(1) phase [from (54.57 +/- 1.19)% to (65.59 +/- 0.41)%, < 0.01] and G(2)/M phase (< 0.05).</p><p><b>CONCLUSION</b>Wortmannin can inhibit proliferation and migration of endothelial cells induced by HGMCM, suggesting that wortmannin carries an anti-angiogenetic ability in diseased retina.</p>

Inhibition of vascular endothelial growth factor gene expression by T7-siRNAs in cultured human retinal pigment epithelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Retinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV.</p><p><b>METHODS</b>Human primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence.</p><p><b>RESULTS</b>Three pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol).</p><p><b>CONCLUSION</b>T7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.</p>

Cloning, expression of soluble VEGFR2 fragment and its effect on tumor angiogenesis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the anti-tumor angiogenesis effect of soluble VEGF receptor fragment by blocking the combination of VEGF and its receptor in vivo and in vitro.</p><p><b>METHODS</b>RT-PCR technique was used to amplify Flk-1/KDR fragment from embryo mouse liver, which was recombinated to expression vector pET-28b(+) and retrovirus vector PLXSN, which was induced to be expressed, purified and identified with EcoR I and Hind III. Mouse endothelial cells were separated, cultured and identified by immunocytochemistrical staining using VIII factor-related antigen antibody. The expressed product was analyzed about its effect on endothelial cell's growth in vitro with MTT method. The retrovirus vector was transfected to tumor cell lines S180 and B16 by liposome method to observe the biological specificity in vitro after gene transfection.</p><p><b>RESULTS</b>1000 bp size sVEGFR fragment was amplify from E9, E11 embryo mouse liver tissues, which was recombinated to TA clone vector and identified by sequence analysis. This fragment was cloned to expression vector pET-28b(+), the expressed product was purified and identified correctly. The in vitro study showed this expressed product can effectively inhibit endothelial cell(s), growth and proliferation. The fragment was then cloned to retrovirus vector PLXSN and transfected to tumor cell lines S180 and B16 successfully with RT-PCR and SDS-PAGE. The experiments in vivo showed that the weight of tumor smaller, the size decreased significantly, the microvessel density was fewer and Flk1 protein expression were higher in the group of gene transfection than that of control.</p><p><b>CONCLUSION</b>Soluble VEGFR fragment is a kind of effective gene engineer product for anti-tumor angiogenesis gene therapy and the development of anti-tumor drug.</p>