Transient reduction of PTI-1 expression by short interfering RNAs inhibits the growth of human prostate cancer cell lines

The prostate tumor-inducing gene 1 (PTI-1) was originally identified by differential ribonucleic acid (RNA) display in human prostate carcinoma. PTI-1 is expressed in human prostate carcinoma but not in benign prostate hypertrophy or normal prostate tissue. PTI-1 may be a member of oncogenes that could affect protein translation and contribute to carcinoma development in human prostate. To investigate the role of PTI-1 in human prostate carcinoma, we constructed three different short interfering RNA (siRNA) vectors (pSilencer3.1-neo-Yu Lei [YL]1-2, -YL3-4 and -YL5-6), each of which was transfected into DU145 and PC3 human prostate cancer cell lines. Among these siRNAs, only pSilencer3.1-neo-YL1-2 could almost completely block the expression of PTI-1 in these two cell lines. The growth of the cell lines was then evaluated after transfection. The proliferation rate was retarded in DU145 and PC3 cells transfected with pSilencer3.1-neo-YL1-2, compared with the cells transfected with a control vector; namely, about 88.6% of DU145 and 80.2% of PC3 cancer cells were blocked at the G1 phase when transfected with pSilencer3.1-neo-YL1-2, compared to 62.0% in DU145 cells and 51.7% in PC3 cells, transfected with the control vector. Moreover, 68.3% of DU145 cells and 72.3% of PC3 cells were induced into apoptosis, while in control transfection, the population was 26.6% in DU145 cells and 28.4% in PC3 cells. These results indicate that blocking PTI-1 expression can inhibit the growth of certain prostate cancer cell lines. We suggest that PTI-1 may serve as a target for the gene-based therapy of human prostate carcinoma.     

Expression of polymeric immunoglobulin receptor mRNA and protein in human paneth cells: Paneth cells participate in acquired immunity

OBJECTIVE: Paneth cells are important effectors of intestinal innate immunity. It has been generally accepted that Paneth cells do not participate in the synthesis of polymeric immunoglobulin receptor (pIgR) or the secretion of immunoglobulin A (IgA) in the small intestine. However, we have previously shown that pIgR is specifically localized in Paneth cells of the rat small intestine. We therefore investigated the possibility that pIgR is also localized in human Paneth cells. METHODS: Double-labeled fluorescent immunohistochemistry and double-labeled fluorescent in situ hybridization were used to determine RNA and protein expression in normal human small intestine. RESULTS: Both pIgR mRNA and protein were colocalized with lysozyme in normal human Paneth cells. Furthermore, IgA was colocalized with lysozyme in the secretory granules of human Paneth cells. CONCLUSIONS: This is the first study to demonstrate that pIgR and IgA are colocalized in the secretory granules of human Paneth cells. These findings suggest that, in addition to their well-recognized role in innate immunity, Paneth cells are involved in IgA-mediated acquired immunity in the gastrointestinal tract. Furthermore, these results add to accumulating evidence that Paneth cells participate in intestinal inflammation.     

Binding activity difference of anti-CD20 scFv-Fc fusion protein derived from variable domain exchange

Two novel engineered antibody fragments binding to antigen CD20 were generated by fusing a murine IgM-type anti-CD20 singie-chain Fv fragment （scFv） to the human IgG1 CH2 （i.e., Cγ2） and CH3 （i.e., Cγ3） domains with the human IgG1 hinge （i.e. Hγ）. Given the relationship between structure and function of protein, the 3-D structures of the two engineered antibody fragments were modeled using computer-aided homology modeling method. Furthermore, the relationship between 3-D conformation and their binding activity was evaluated theoretically. Due to the change of active pocket formed by CDRs, the HL23 （VH-Linker-VL-Hγ-Cγ2-Cγ3） remained its activity because of its preserved conformation, while the binding activity of the LH23 （VL-Linker-VH-Hγ-Cγ2-Cγ3） was impaired severely. Experimental studies by flow cytometry and fluorescence microscopy showed that HL23 possessed significantly superior binding activity to CD20-expressing target cells than LH23. That is to say, the order of variable regions could influence the binding activity of the fusion protein to CD20^＋ cell lines, which was in accordance with the theoretical results. The study highlights the potential relationship between the antibody binding activity and their 3-D conformation, which appears to be worthwhile in providing direction for future antibody design of recombinant antibody.     

Dual-probe assay for detection of lamivudine-resistance hepatitis B virus by real-time PCR

A rapid and accurate minor groove binder (MGB) real-time PCR is described for detection lamivudine resistance mutations in hepatitis B virus. The real-time PCR was compared with direct Sanger sequencing in 53 clinical patients samples, the results of the real-time PCR correlate with the nucleotide sequence and the assay has the advantage of detecting a mixture of quasi-species with higher sensitivity than sequencing. As a simple, easy, rapid, accurate and high throughout method, MGB real-time PCR assay should be useful for detecting lamivudine resistance variants during lamivudine therapy in clinical settings.     

Internalization of Jembrana disease virus Tat: possible pathway and implication

Jembrana disease virus (JDV) is a lentivirus highly related to the bovine immunodeficiency virus (BIV). It causes an acute disease with high mortality rate within 1-2 weeks. JDV encodes the most potent Tat (JTat) of any of the lentiviruses. JTat can transactivate all LTRs and functionally substitute for HIV Tat in the viral genome and may function as a pivotal regulator in the acute pathogenesis of JDV. The goal of this paper is to study JTat internalization by cells, the mechanisms involved in internalization, and the effect of JTat on neighbouring cells. By quantification and fluorescence microscopy, we found that the internalization of extracellular EGFP-JTat fusion protein was both time and dose-dependent, but endocytosis and energy independent. We identified that arginines which were responsible for the internalization. Internalized JTat was distributed in both the nucleus and the cytoplasm, could transactivate JDV LTR and modulate cellular gene expression. Based on our findings, we propose that secretion and internalization of JTat may be a way for JDV to influence neighbouring cells and make the cellular environment more amenable to viral infection.     

A photolithographic method to create cellular micropatterns

Here we describe a simple and rapid system for creation of patterned cell culture substrates. This technique is based on (1) printing a mask on a standard overhead transparency, (2) coating a thin layer of a photocrosslinkable chitosan on a slide, (3) exposing the slide and mask to ultraviolet (UV) light, and (4) rinsing the uncrosslinked polymer to expose the underlying cell-repellent patterns. Photocross-linkable chitosan does not require photoinitiators, it is non-toxic and forms flexible, biocompatible hydrogel upon short ( approximately min) UV exposure. Patterns of various shapes (lanes, squares, triangles, circles) were created on two surfaces commonly used for cell culture: glass and tissue culture polystyrene. The pattern size could be varied with a mum resolution using a single mask and varying UV exposure time. Cardiac fibroblasts formed stable patterns for up to 18 days in culture. Cardiomyocytes, patterned in lanes 68-99 microm wide, exhibited expression of cardiac Troponin I, well developed contractile apparatus and they contracted synchronously in response to electrical field stimulation. Osteoblasts (SAOS-2) localized in the exposed glass regions (squares, triangles, or circles; 0.063-0.5mm(2)). They proliferated to confluence in 5 days, expressed alkaline phosphatase and produced a mineralized matrix.     

Application of a new dosimetry program TAOCS to assess transient vapour absorption in the upper airways

Most previous models of vapour absorption in the respiratory tract have assumed steady state flow fields and steady state diffusion into the airway walls. However, recent studies have shown that transient absorption flux into the walls of the upper airways can significantly influence predicted uptake or deposition values. The disadvantage of accounting for transient absorption into the airway walls is a more complex boundary condition and numerical model. The objective of this study was to evaluate the effects of both transient flow fields and transient mass absorption on the uptake of highly and moderately soluble compounds in an upper airway model. The geometry consisted of the mouth-throat region coupled with a multilayer wall model containing air, mucus, tissue, and blood phases. Based on previous studies, a boundary condition that represents transient absorption into the airway walls was applied. A new dosimetry program, named transient absorption of chemical species (TAOCS) 1.0, was developed and implemented to determine the coefficients needed for the transient boundary condition expression and to apply the boundary condition to the computational fluid dynamics (CFD) model. Both steady state and transient conditions were considered for the airflow field and wall absorption. The case of perfect wall absorption with a zero surface concentration was also considered. Results indicated that steady state airflow provided a reasonable approximation to transient airflow conditions in terms of total and local deposition (values within 10-30%). However, the simulation of transient wall absorption was critical unless the compound was highly soluble (with a mucus-air partition coefficient ≥320), in which case a perfect absorption boundary condition was accurate to within a relative difference of 50%. Still, the perfect absorption boundary condition did not accurately capture local deposition enhancement factor values. Based on these findings, implementation of the transient absorption boundary condition appears critical to predict local deposition characteristics for even highly soluble compounds. Use of the TAOCS program simplified the implementation of the complex transient absorption condition making the CFD simulation process more efficient and user-friendly.     

伴恶性胸腔积液Ⅳ期NSCLC原发肿瘤三维放疗的临床分析

目的 回顾性探讨伴恶性胸腔积液Ⅳ期NSCLC原发灶三维放疗的有效性和安全性。方法 选择2007-2018年经病理诊断、初诊的伴恶性胸腔积液Ⅳ期NSCLC198例,按治疗状况分为未治疗组45例、药物治疗组57例、放疗组96例。分析放疗组和药物组近期疗效、总生存和不良反应。Kaplan-Meier法计算生存率并log-rank检验,Cox模型多因素预后分析。结果 放疗组有效率为54%、无效率为46%,显著优于药物组的25%、75%(P=0.007);1、2、3、5年总生存和中位生存期放疗组分别为47%、18%、6%、1%和12个月,显著高于药物组的15%、3%、2%、0%和5个月(P<0.001)。多因素分析显示增加原发灶放疗是延长生存的影响因素(P<0.001), ≥63Gy、4~6周期化疗有延长生存趋势(P=0.063、0.071),EGFR敏感突变者放疗联合分子靶向治疗总生存显著优于联合化疗而EGFR突变未知者(P=0.007)。增加原发灶放疗未增加明显的不良反应(P>0.05)。结论 伴恶性胸腔积液Ⅳ期NSCLC原发灶三维放疗可能延长患者总生存,增加三维放疗的不良反应可耐受。     

Unrelated cord blood transplantation for central nervous system relapse in high-risk childhood acute lymphoblastic leukemia

Few clinical studies have investigated the role of unrelated cord blood transplantation (CBT) for central nervous system (CNS) relapse of childhood acute lymphoblastic leukemia (ALL) patients with high-risk factors. The aim of this report is to identify the potential benefits of unrelated CBT in high-risk childhood ALL with CNS relapse who has been treated on CNS-directed treatment strategies. Eleven childhood ALL patients with CNS relapse who underwent unrelated CBT enrolled in our study between 2001 and 2011, and all of the patients had features associated with poor outcomes, such as high white blood cells at diagnosis, ph?+?chromosome, or a history of bone marrow relapse. All transplants were performed with myeloablative-conditioning therapy (BU/cyclophosphamide (CY₂) or total body irradiation/CY) plus highly CNS-active agents (carmustine or high-dose cytarabine). All patients achieved neutrophil engraftment and platelet engraftment. A total of nine patients (81.8 %) developed pre-engraftment syndrome at a median of 7 days, and three patients developed acute graft-vs-host disease at a median of 21 days. The median follow-up after CBT was 28.5 months. The probability of overall survival at 9 years was 63.6 %, and no patient experienced a CNS relapse. Our experience suggests that unrelated CBT appears to be an effective treatment option for CNS relapse of childhood ALL patients associated with poor outcome features.     

Angiotensin converting enzyme inhibitors for prevention of new-onset type 2 diabetes mellitus: A meta-analysis of 72,128 patients

Backgroud

Angiotensin converting enzyme inhibitors (ACEIs) have been linked to reduced risk of new-onset diabetes, but the evidence was insufficient.

Objective and methods

The aim of this study was to evaluate the effect of ACEIs on the development of new-onset type 2 diabetes. Randomized controlled trials (RCTs) about ACEIs and new-onset diabetes were identified by electronic and manual searches.

Results

Nine RCTs with 92,404 patients (72,128 non-diabetic patients at baseline) were included in this study. Compared with control group, incidence of new-onset diabetes was significantly reduced in the ACEIs group [OR 0.80, (0.71, 0.91)], irrespective of achieved blood pressure levels at the follow-up. ACEIs therapy was associated with significant reduction in the risk of new-onset diabetes compared with beta-blockers/diuretics [OR 0.78, (0.65, 0.93)], placebo [OR 0.79, (0.64, 0.96)], or calcium channel blockers [OR 0.85, (0.73, 0.99)]. ACEIs treatment was associated with significant reduction in the risk of new-onset diabetes in patients with hypertension [OR 0.80, (0.68, 0.93)], coronary artery disease (CAD) or cardiovascular disease [OR 0.83, (0.68, 1.00)], or heart failure [OR 0.22, (0.10, 0.47)]. Among patients with impaired glucose tolerance or impaired fasting glucose, ramipril did not significantly reduce the incidence of diabetes [OR 0.91, (0.79, 1.05)], but significantly increased regression to normoglycemia.

Conclusion

ACEIs have beneficial effects in preventing new-onset diabetes. ACEIs provide additional benefits of lowering the risk of new-onset diabetes in patients with hypertension, CAD or other cardiovascular disease.