民族植物学定量研究中的取样方法

定量方法在民族植物学研究中越来越占有重要的地住,但民族植物学定量研究中有关取样方面还存在着一些不同的看法。笔者针对云南省金平苗族瑶族傣族自治县的拉祜族民间草医所使用的药用植物资源和草医对不同生境中药用植物的利用情况进行了取样,并以此对民族植物学中有关取样中存在的问题进行了讨论。结果表明不同生境中民族植物学的取样面积与植物生态学中的取样面积比较相近。所以在进行民族植物学野外取样时,司以针对具体的生境和植被特点来借用植物生态学中对该类生境或植被所采用的最小取样面积。     

Accumulation and tolerance characteristics of lead in <Emphasis Type="Italic">Althaea rosea</Emphasis> Cav. and <Emphasis Type="Italic">Malva crispa</Emphasis> L.

Two ornamental plants of Althaea rosea Cav. and Malva crispa L. were exposed to various concentrations of lead (Pb) (0, 50, 100, 200 and 500 mg·kg^?1) for 70 days to evaluate the accumulating potential and the tolerance characteristics. The results showed that both plant species grown normally under Pb stress, and A. rosea had a higher tolerance than M. crispa, while M. crispa had a higher ability in Pb accumulation than A. rosea. Besides, lower Pb concentration (50 mg·kg^?1) stimulated the shoot biomass in both plant species. Pb accumulation in plants was consistent with the increase of Pb levels, and the main accumulation sites were the roots and the older leaves. In addition, the photosynthetic pigments content and chlorophyll fluorescence parameters were influenced by Pb stress. In such case, both of the plants could improve the activities of antioxidant enzymes of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), and the contents of the total soluble sugar and soluble protein, which reached the highest value at Pb 100 mg·kg^?1, as well as the accumulation of the total thiols (T-SH) and non-protein thiols (NP-SH) to adapt to Pb stress. Thus, it provides the theoretical basis and possibility for ornamental plants of A. rosea and M. crispa in phytoremediation of Pb contaminated areas.     

Long non‐coding RNA AFAP1‐AS1/miR‐320a/RBPJ axis regulates laryngeal carcinoma cell stemness and chemoresistance

AFAP1‐AS1 is a long non‐coding RNA that is associated with tumorigenesis and poor prognosis in a variety of cancers. We have been suggested that AFAP1‐AS1 increases tumorigenesis in laryngeal carcinoma specifically by enhancing stemness and chemoresistance. We assessed AFAP1‐AS1 expression in human laryngeal specimens, paired adjacent normal tissues and human HEp‐2 cells. Indeed, we found not only that AFAP1‐AS1 was up‐regulated in laryngeal carcinoma specimens and cells, but also that stemness‐associated genes were overexpressed. Silencing of AFAP1‐AS1 promoted HEp‐2 cell chemoresistance under cisplatin treatment. Expression of AFAP1‐AS1 was increased in drug‐resistant Hep‐2 cells. We then probed the mechanism of AFAP1‐AS1 activity and determined that miR‐320a was a potential molecular target of AFAP1‐AS1. Luciferase reporter and qRT‐PCR assays of AFAP1‐AS1 and miR‐320a levels in human specimens and cell cultures indicated that AFAP1‐AS1 negatively regulates miR‐320a. To discover the molecular mechanism of miR‐320a, we again used the DIANA Tools algorithm to predict its genetic target, RBPJ. After cloning the 3′‐untranslated regions (3′‐UTR) of RBPJ into a luciferase reporter, we determined that miR‐320a did in fact reduce RBPJ mRNA and protein levels. Ultimately, we determined that AFAP1‐AS1 increases RBPJ expression by negatively regulating miR‐320a and RBPJ overexpression rescues stemness and chemoresistance inhibited by AFAP1‐AS1 silencing. Taken together, these results suggest that AFAP1‐AS1 can serve as a prognostic biomarker in laryngeal carcinoma and that miR‐320a has the potential to improve standard therapeutic approaches to the disease, especially for cases in which cancer cell stemness and drug resistance present significant barriers to effective treatment.     

Clinical Correlates of Mass Effect in Autosomal Dominant Polycystic Kidney Disease

Mass effect from polycystic kidney and liver enlargement can result in significant clinical complications and symptoms in autosomal dominant polycystic kidney disease (ADPKD). In this single-center study, we examined the correlation of height-adjusted total liver volume (htTLV) and total kidney volume (htTKV) by CT imaging with hepatic complications (n = 461) and abdominal symptoms (n = 253) in patients with ADPKD. “Mass-effect” complications were assessed by review of medical records and abdominal symptoms, by a standardized research questionnaire. Overall, 91.8% of patients had 4 or more liver cysts on CT scans. Polycystic liver disease (PLD) was classified as none or mild (htTLV < 1,600 mL/m); moderate (1,600 ≤ htTLV <3,200 mL/m); and severe (htTLV ≥ 3,200 mL/m). The prevalence of moderate and severe PLD in our patient cohort was 11.7% (n = 54/461) and 4.8% (n = 22/461), respectively, with a female predominance in both the moderate (61.1%) and severe (95.5%) PLD groups. Pressure-related complications such as leg edema (20.4%), ascites (16.6%), and hernia (3.6%) were common, and patients with moderate to severe PLD exhibited a 6-fold increased risk (compared to no or mild PLD) for these complications in multivariate analysis. Similarly, abdominal symptoms including back pain (58.8%), flank pain (53.1%), abdominal fullness (46.5%), and dyspnea/chest-discomfort (44.3%) were very common, and patients with moderate to severe PLD exhibited a 5-fold increased risk for these symptoms. Moderate to severe PLD is a common and clinically important problem in ~16% of patients with ADPKD who may benefit from referral to specialized centers for further management.     

Effects of small interfering RNAs targeting MAPK1 on gene expression profile in HeLa cells as revealed by microarray analysis

The mitogen activated protein kinases (MAPK) signaling cascade plays an important role in cell life. We proved that small interfering RNAs targeting MAPK1 (siRNA-2) could inhibit HeLa cell growth, but the effects of siRNA-2 on gene expression profile were unclear. Using Affymetrix GeneChip HG-U133A 2.0, we identified the long-term changes for 48h in gene expression profile in HeLa cell treated by siRNA-2. The results showed that expressions of 181 genes were altered by siRNA-2 and were divided into two groups: (i) one group showed downregulation by siRNA-2, including the proliferation associated genes, small G protein, cytoskeleton associated protein and extracellular matrix associated protein; and (ii) the other group showed upregulation by siRNA-2, including interferon response genes, OAS family, TRIM family and apoptosis associated genes. The results of Real-time quantitative PCR for MAPK1, NUP188, P38, STAT1, STAT2, MPL and OAS1 were consistent with that of gene chip. Two networks were found to react substantially to the downregulation of MAPK1 by siRNA-2. One of the networks regulates cell growth through cell-cycle control, apoptosis and cytoskeleton. The other network is related to interferon-like response. Our findings suggest that siRNA-mediated downregulation of MAPK1 could be an attractive strategy for cancer therapy.     

RPE barrier breakdown in diabetic retinopathy: seeing is believing

Diabetic retinopathy (DR) is a major complication of diabetes and a leading cause of blindness in working-age Americans. DR is traditionally regarded as a disorder of blood–retina barriers, and the leakage of blood content is a major pathological characteristic of the disease. While the breakdown of the endothelial barrier in DR has been investigated extensively, the vascular leakage through the retinal pigment epithelium (RPE) barrier in the disease has not been widely acknowledged. As the blood content leaked through the RPE barrier causes excessive water influx to the retina, the breakdown of the RPE barrier is likely to play a causative role in the development of some forms of diabetic macular edema, a major cause of vision loss in DR. In this article, we will discuss the clinical evidences of the diabetes-induced RPE barrier breakdown, the alteration of the RPE in diabetes, the molecular and cellular mechanism of RPE barrier breakdown, and the research tools for the analysis of RPE barrier leakage. Finally, we will discuss the methodology and potential applications of our recently developed fluorescent microscopic imaging for the diabetes- or ischemia-induced RPE barrier breakdown in rodents.     

陕西食源性沙门氏菌耐药及相关基因

【目的】研究食源性沙门氏菌对常用抗生素的药敏性及相关耐药基因,更好的了解耐药性的产生和传播途径,确保食品安全。【方法】使用the Clinical and Laboratory Standards Institute推荐的琼脂稀释法测定沙门氏菌的药敏性,PCR和基因序列测定方法确定耐药沙门氏菌中整合子及其携带的耐药基因、与头孢菌素抗性相关的基因、沙门氏菌基因岛及与氟喹诺酮类抗生素耐药相关的基因突变。【结果】359株沙门氏菌中,67%的菌株对磺胺甲恶唑产生抗性,对甲氧苄啶/磺胺甲恶唑、四环素、卡那霉素、萘啶酮酸、氨苄西林、阿莫西林/克拉维酸、链霉素、氯霉素和庆大霉素、环丙沙星、头孢曲松、头孢西丁和头孢哌酮的耐药率分别为58%、56%、37%、35%、33%、32%、29%、26%、21%、16%、9%和8%。284株耐药菌中,79%的菌株可抗至少1种抗生素,25.9%可抗10种以上抗生素,2.5%可抗14种抗生素。耐药的Ⅰ类整合子以1.4kb最为常见,携带的耐药基因有aadA1、aadA2、aadA5、tetR、blaPSE-1、blaDHA-1、blaVEB-1、dhfrⅠ、dhfrⅤ、dhfrⅦ和dhfr17等。62株耐头孢曲松和/或头孢哌酮的沙门氏菌中,blaTEM和blaCMY-2基因的检出率分别为51.6%和56.5%。13.6%的沙门氏菌中检出了沙门氏菌基因岛。35株耐氟喹诺酮类抗生素的沙门氏菌的gyrA、parC和parE基因中共检出68个点突变,gyrA基因中常见突变为Ser83Phe、Ser83Tyr、Asp87Gly和Asp87Asn,parC基因中为Ser80Arg。parE基因中检出了Lys441Ile、Lys428Gln、Asp494Asn、Lys428Gln和Gly442Ser突变,这些点突变均为首次在食源性沙门氏菌中检出。【结论】陕西食源性沙门氏菌耐药状况严重,整合子、沙门氏菌基因岛和β-内酰胺酶编码基因的存在及解旋酶和拓扑异构酶基因突变是导致沙门氏菌耐药的重要机制。     

球孢白僵菌高渗适应性相关基因Bbmpd的克隆与表达分析

【目的】克隆与球孢白僵菌(Beauveria bassiana)的高渗适应性相关基因,并对其功能进行分析,以揭示球孢白僵菌对高渗等逆境适应的分子机理。【方法】利用YADE法克隆T-DNA的侧翼序列并进行基因组步行,获得突变基因的全长及上游序列;利用RT-PCR技术分析突变基因的表达特性以及与Bbhog1的关系;采用同源重组技术敲除Bbmpd基因。【结果】克隆得到插入突变基因及其上、下游序列全长3037bp。该基因与编码球孢白僵菌的1-磷酸甘露醇脱氢酶基因相似性为98%。Bbmpd的表达受高渗环境(0.8mol/L NaCl)的诱导,受Bbhog1信号途径的激活调节,Bbhog1缺失导致Bbmpd表达下调。Bbmpd缺失突变体在高渗胁迫下的生长受到明显抑制。Bbmpd缺失不影响球孢白僵菌在查氏培养基上的生长和产孢。【结论】由T-DNA突变体克隆了编码球孢白僵菌1-磷酸甘露醇脱氢酶基因Bbmpd,该基因的表达受高渗环境的诱导和Bbhog1的调控,与球孢白僵菌高渗适应性相关。     

Putative phosphorylation sites on WCA domain of HA2 is essential for <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> single nucleopolyhedrovirus replication

Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality.The Helicoverpa armigera single nucleopolyhedrovirus (HearNPv) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain,in which phosphorylation status are supposed to be critical in respect to actin polymerization.In the present study,two putative phosphorylation sites (232Thr and 250Ser) and a highly conserved Serine (245Ser) on the WCA domain of HA2 were mutated,and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome.Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at 245Ser can produce infectious virions,both 232Tbr and 250Ser mutations were lethal to the virus.However,actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus,which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway.