人乳头状瘤病毒DNA检测在宫颈病变筛查中的应用研究

目的：探讨高危型人乳头状瘤病毒（HR-HPV）DNA检测方法在宫颈病变筛查中的应用意义。方法：580例妇女同时进行薄层液基细胞学（TCT）、第2代杂交捕获法（HC Ⅱ）和阴道镜下宫颈组织活检,并以病理组织学检查结果作为确诊标准进行对比分析。结果：①580例受检者中病理诊断为炎症207例（35.69%）,CIN Ⅰ 224例（38.62%）,CIN Ⅱ 96例（16.55%）,CIN Ⅲ 38例（6.55%）,浸润癌15例（2.58%）;②TCT检测异常者中炎症52例（25.12%）,CIN Ⅰ 177例（79.02%）,CIN Ⅱ 85例（88.54%）,CIN Ⅲ36例（94.74%）,浸润癌15例（100%）,其中CIN Ⅱ和CIN Ⅲ组间差异无统计学意义（P〉0.05）,但显著高于炎症组和CIN Ⅰ组,低于湿润癌组（P〈0.01或0.05）;③HPV DNA检测阳性者中炎症66例（31.88%）,CIN Ⅰ 152例（67.86%）,CIN Ⅱ 83例（86.46%）,CINIII 35例（92.11%）,浸润癌组15例（100%）,除CIN Ⅱ和CIN Ⅲ组间差异无显著性外（P〉0.05）,其余各组间差异均有统计学意义（P〈0.05或0.01）,且HPV-DNA检测阳性组CIN和浸润癌发病率明显高于阴性组（P〈0.01）;④30岁以下高危险型HPV感染率（65.53%）显著高于30岁以上34.47%感染率（P〈0.01）;⑤联合应用TCT、HPV-DNA检测诊断宫颈癌及癌前病变的敏感度和特异度分别为96.14%和69.28%,高于TCT或HPV-DNA的单独检测。结论：宫颈高危险型HPV感染是CIN及宫颈癌的主要发病因素,并与病变严重程度密切相关,而HPV-DNA和TCT联合应用可提高宫颈癌及癌前病变的检出率。     

胃转流术后远端肠黏膜形态及生长因子表达的变化

目的：探讨胃转流术后远端肠管黏膜的适应性变化及生长因子的表达情况。方法：将8周龄的Wistar大鼠随机分为对照组、假手术组和胃转流术组,术后8周取吻合口远端肠管行常规病理切片检查,测量肠黏膜厚度和绒毛高度,采用免疫荧光法检测肠粘膜中表皮生长因子（EGF）和胰岛素样生长因子-1（IGF—1）的表达。结果：胃转流术组黏膜厚度（672±39与500±31um,P〈0．01）和绒毛高度（445±19与342±15um,P〈0．01）均显著高于假手术组（P〈0．01）,而对照组和假手术组间均无显著差异。与假手术组比较,胃转流术组肠粘膜中EGF和IGF-1的表达显著升高（P〈0．01）,而对照组和假手术组间无显著差异。结论：胃转流术后远端肠管粘膜发生适应性增生,同时伴随着EGF和IGF—1表达水平的升高。     

Proteomic identification of IPSE/alpha-1 as a major hepatotoxin secreted by Schistosoma mansoni eggs

Background

Eggs deposited in the liver of the mammalian host by the blood fluke parasite, Schistosoma mansoni, normally drive a T-helper-2 (Th2)-mediated granulomatous response in immune-competent mice. By contrast, in mice deprived of T-cells and incapable of producing granulomata, egg-secreted proteins (ESP) induce acute hepatic injury and death. Previous work has shown that one such ESP, the T2 ribonuclease known as omega-1, is hepatotoxic in vivo in that specific antisera to omega-1 prevent hepatocyte damage.

Methodology/Principal Findings

Using an in vitro culture system employing mouse primary hepatocytes and alanine transaminase (ALT) activity as a marker of heptocyte injury, we demonstrated that S. mansoni eggs, egg-secreted proteins (ESP), soluble-egg antigen (SEA), and omega-1 are directly hepatotoxic and in a dose-dependent manner. Depletion of omega-1 using a monoclonal antibody abolished the toxicity of pure omega-1 and diminished the toxicity in ESP and SEA by 47 and 33%, respectively. Anion exchange chromatography of ESP yielded one predominant hepatotoxic fraction. Proteomics of that fraction identified the presence of IPSE/alpha-1 (IL-4 inducing principle from S. mansoni eggs), a known activator of basophils and inducer of Th2-type responses. Pure recombinant IPSE/alpha-1 also displayed a dose-dependent hepatotoxicity in vitro. Monoclonal antibody depletion of IPSE/alpha-1 abolished the latter''s toxicity and diminished the total toxicity of ESP and SEA by 32 and 35%, respectively. Combined depletion of omega-1 and IPSE/alpha-1 diminished hepatotoxicity of ESP and SEA by 60 and 58% respectively.

Conclusions

We identified IPSE/alpha-1 as a novel hepatotoxin and conclude that both IPSE/alpha-1 and omega-1 account for the majority of the hepatotoxicity secreted by S. mansoni eggs.     

Altered insulin receptor signalling and β-cell cycle dynamics in type 2 diabetes mellitus

Insulin resistance, reduced β-cell mass, and hyperglucagonemia are consistent features in type 2 diabetes mellitus (T2DM). We used pancreas and islets from humans with T2DM to examine the regulation of insulin signaling and cell-cycle control of islet cells. We observed reduced β-cell mass and increased α-cell mass in the Type 2 diabetic pancreas. Confocal microscopy, real-time PCR and western blotting analyses revealed increased expression of PCNA and down-regulation of p27-Kip1 and altered expression of insulin receptors, insulin receptor substrate-2 and phosphorylated BAD. To investigate the mechanisms underlying these findings, we examined a mouse model of insulin resistance in β-cells--which also exhibits reduced β-cell mass, the β-cell-specific insulin receptor knockout (βIRKO). Freshly isolated islets and β-cell lines derived from βIRKO mice exhibited poor cell-cycle progression, nuclear restriction of FoxO1 and reduced expression of cell-cycle proteins favoring growth arrest. Re-expression of insulin receptors in βIRKO β-cells reversed the defects and promoted cell cycle progression and proliferation implying a role for insulin-signaling in β-cell growth. These data provide evidence that human β- and α-cells can enter the cell-cycle, but proliferation of β-cells in T2DM fails due to G1-to-S phase arrest secondary to defective insulin signaling. Activation of insulin signaling, FoxO1 and proteins in β-cell-cycle progression are attractive therapeutic targets to enhance β-cell regeneration in the treatment of T2DM.     

Chauhanellus Bychowsky & Nagibina, 1969 (Monogenea) from ariid fishes (Siluriformes) of Peninsular Malaysia

Twelve new species of Chauhanellus Bychowsky & Nagibina, 1969 have been found on six species of ariid from Peninsular Malaysia: Chauhanellus trifidus n. sp., C. digitalis n. sp., C. malayanus n. sp., C. forcipis n. sp. and C. intermedius n. sp. from Arius sagor; C. aspinous n. sp. from Arius venosus; C. caelatus n. sp. from Arius caelatus; C. auriculatum n. sp., C. poculus n. sp. and C. pulutanus n. sp. from Arius maculatus; C. duriensis n. sp. from Arius thalassinus; and C. osteogeneiosi n. sp. from Osteogeneiosus militaris. Some of these Chauhanellus species possess characteristics that are not commonly observed in the genus. C. aspinous n. sp., C. intermedius n. sp. and C. digitalis n. sp. exhibit features found in both Chauhanellus and Hamatopeduncularia: these include absence of spines on the mainpart of the dorsal anchors in C. aspinous n. sp. and C. intermedius n. sp. and presence of haptoral digitation in C. digitalis n. sp. Other features are the five transverse rows of peduncular spines in C. duriensis n. sp., ear-like projections on the anchors in C. auriculatum n. sp., and thin sclerotised plates that partly envelope the ventral anchors in C. forcipis n. sp. Mid-dorsal appendices occur on the dorsal bars of seven of the present species.     

The histone demethylase Jmjd3 regulates zebrafish myeloid development by promoting spi1 expression

The histone demethylase Jmjd3 plays a critical role in cell lineage specification and differentiation at various stages of development. However, its function during normal myeloid development remains poorly understood. Here, we carried out a systematic in vivo screen of epigenetic factors for their function in hematopoiesis and identified Jmjd3 as a new epigenetic factor that regulates myelopoiesis in zebrafish. We demonstrated that jmjd3 was essential for zebrafish primitive and definitive myelopoiesis, knockdown of jmjd3 suppressed the myeloid commitment and enhanced the erythroid commitment. Only overexpression of spi1 but not the other myeloid regulators rescued the myeloid development in jmjd3 morphants. Furthermore, preliminary mechanistic studies demonstrated that Jmjd3 could directly bind to the spi1 regulatory region to alleviate the repressive H3K27me3 modification and activate spi1 expression. Thus, our studies highlight that Jmjd3 is indispensable for early zebrafish myeloid development by promoting spi1 expression.     

A thermostable shikimate 5-dehydrogenase from the archaeon Archaeoglobus fulgidus

Shikimate 5-dehydrogenase (SKDH; EC 1.1.1.25) catalyzes the reversible reduction of 3-dehydroshikimate to shikimate and is a key enzyme in the aromatic amino acid biosynthesis pathway. The shikimate 5-dehydrogenase gene, aroE, from Archaeoglobus fulgidus was cloned and overexpressed in Escherichia coli. The recombinant enzyme purified as a homodimer and yielded a maximum specific activity of 732 U/mg at 87 degrees C (with NADP+ as coenzyme). Apparent Km values for shikimate, NADP+, and NAD+ were estimated at 0.17+/-0.03 mM, 0.19+/-0.01 mM, and 11.4+/-0.4 mM, respectively. The half-life of the A. fulgidus SKDH is 2 h at the assay temperature (87 degrees C) and 17 days at 60 degrees C. Addition of 1 M NaCl or KCl stabilized the enzyme's half-life to approximately 70 h at 87 degrees C and approximately 50 days at 60 degrees C. This work presents the first kinetic analysis of an archaeal SKDH.     

Six2 is required for suppression of nephrogenesis and progenitor renewal in the developing kidney

During kidney development and in response to inductive signals, the metanephric mesenchyme aggregates, becomes polarized, and generates much of the epithelia of the nephron. As such, the metanephric mesenchyme is a renal progenitor cell population that must be replenished as epithelial derivatives are continuously generated. The molecular mechanisms that maintain the undifferentiated state of the metanephric mesenchymal precursor cells have not yet been identified. In this paper, we report that functional inactivation of the homeobox gene Six2 results in premature and ectopic differentiation of mesenchymal cells into epithelia and depletion of the progenitor cell population within the metanephric mesenchyme. Failure to renew the mesenchymal cells results in severe renal hypoplasia. Gain of Six2 function in cortical metanephric mesenchymal cells was sufficient to prevent their epithelial differentiation in an organ culture assay. We propose that in the developing kidney, Six2 activity is required for maintaining the mesenchymal progenitor population in an undifferentiated state by opposing the inductive signals emanating from the ureteric bud.     

Tick-Borne Rickettsial Pathogens in Ticks and Small Mammals in Korea

In order to investigate the prevalence of tick-borne infectious agents among ticks, ticks comprising five species from two genera (Hemaphysalis spp. and Ixodes spp.) were screened using molecular techniques. Ticks (3,135) were collected from small wild-caught mammals or by dragging/flagging in the Republic of Korea (ROK) and were pooled into a total of 1,638 samples (1 to 27 ticks per pool). From the 1,638 tick samples, species-specific fragments of Anaplasma phagocytophilum (1 sample), Anaplasma platys (52 samples), Ehrlichia chaffeensis (29 samples), Ehrlichia ewingii (2 samples), Ehrlichia canis (18 samples), and Rickettsia rickettsii (28 samples) were amplified by PCR assay. Twenty-one pooled and individual tick samples had mixed infections of two (15 samples) or three (6 samples) pathogens. In addition, 424 spleen samples from small captured mammals (389 rodents, 33 insectivores, and 2 weasels) were screened for selected zoonotic pathogens. Species-specific DNA fragments of A. phagocytophilum (110 samples), A. platys (68 samples), E. chaffeensis (8 samples), E. ewingii (26 samples), E. canis (51 samples), and Rickettsia sp. (22 samples) were amplified by PCR assay. One hundred thirty small mammals had single infections, while 4, 14, and 21 striped field mice (Apodemus agrarius) had mixed infections of four, three, and two pathogens, respectively. Phylogenetic analysis based on nucleotide sequence comparison also revealed that Korean strains of E. chaffeensis clustered closely with those from China and the United States, while the Rickettsia (rOmpA) sequences clustered within a clade together with a Chinese strain. These results suggest that these agents should be considered in differential diagnosis while examining cases of acute febrile illnesses in humans as well as animals in the ROK.