Sequence of the Human Iduronate 2-Sulfatase (IDS) Gene

Deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS; EC 3.1.6.13) results in the storage of the glycosaminoglycans heparan sulfate and dermatan sulfate, which leads to the lysosomal storage disorder mucopolysaccharidosis type II. Three overlapping genomic clones derived from an X-chromosome-specific library containing the entire IDS gene were isolated and the sequences of the intron boundaries and the 5′ promoter region were determined. The IDS gene is split into nine exons spanning approximately 24 kb. The potential promoter for IDS lacks a TATA box but contains GC box consensus sequences, consistent with its role as a housekeeping gene. A polypyrimidine-like repeat is found in intron 1.     

Stimulation of Human Hair Growth by the Recombinant Human Keratinocyte Growth Factor-2 (KGF-2)

Keratinocyte growth factor-2 (KGF-2) is found in dermal papilla fibroblasts and its receptor, fibroblast growth factor receptor 2 (FGFR2), in the neighboring outer root sheath of keratinocytes. Administration of recombinant human KGF-2 (rhKGF-2) at 10 ng ml⁻¹ significantly stimulated human hair-follicle cell proliferation in organ culture (26–35%). Thus, rhKGF-2 is a promising therapeutic agent to stimulate human hair growth.     

重组人生长激素治疗重型肝炎的疗效观察

目的：观察重组人生长激素治疗重型肝炎的疗效。方法：回顾性分析我院住院治疗的60例重型肝炎患者并分组比较,对照组25例为包括血浆置换在内的综合内科治疗,治疗组35例为在对照组基础上联合重组人生长激素治疗;分析两组治疗前后的临床症状、体征和肝肾功能、凝血功能等指标变化,并对治疗结果进行分析,评价临床疗效。结果：治疗组临床症状改善有效率为93.3％,高于对照组（72．0％）（P〈0．05）;观察组肝功能指标好转率80．0％,优于对照组65．5％（P〈0．05）。结论：重组人生长激素治疗重型肝炎有较好的效果。     

重组人生长激素的主要药效学研究

参照文献方法,观察了ｒｈＧＨ对正常大鼠（３月龄,平台期）生长发育作用的影响,ｉｍ,ｒｈＧＨ（０．０５、０．１５、０．４５ｍｇ．ｋｇ＾－１．ｄ＾－１）,连续１５天,结果表明：ｒｈＧＨ（０．１５、０．４５ｍｇ．ｋｇ＾－１．ｄ＾－１）同阳性对照生长激素ＮＩＢＳＣ标准品）一样,均可明显促进正常平台期大鼠的体重及尾长增加,大剂量组还可明显促进肝、肾及骨骼的生长发育,使肝、肾重量增加,胫骨骨骺端距离增大,同     

Genetic Analysis of 17 Children with Hunter Syndrome:Identification and Functional Characterization of Four Novel Mutations in the Iduronate-2-Sulfatase Gene

Mucopolysaccharidosis type II （MPS II） is a rare X-linked disorder caused by alterations in the iduronate-2-sulfatase （IDS） gene. In this study, IDS activity in peripheral mononuclear blood monocytes （PMBCs） was measured with a fluorimetric enzyme assay. Urinary glycosaminoglycans （GAGs） were quantified using a colorimetric assay. All IDS exons and intronic flanks were bidirectionally sequenced. A total of 15 mutations （all exonic region） were found in 17 MPS II patients. In this cohort of MPS II patients, all alterations in the IDS gene were caused by point nucleotide substitutions or small deletions. Mutations p.Arg88His and p.Arg172＊ occurred twice. All mu- tations were inherited except for p.Gly489Alafs＊7, a germline mutation. We found four new mutations （p.Ser142Phe, p.Arg233Gly, p.Glu430＊, and p.Ile360Tyrfs＊31）. In Epstein-Barr virus （EBV）-immortalized PMBCs derived from the MPS II patients, no IDS protein was detected in case of the p.Ser142Phe and p.Ile360Tyrfs＊31 mutants. For p.Arg233Gly and p.Glu430＊, we observed a residual expression of IDS. The p.Arg233Gly and p.Glu430＊ mutants had a residuary enzymatic activity that was lowered by 14.3 and 76-fold, respectively, compared with healthy controls. This observation may help explain the mild disease phenotype in MPS II patients who had these two mutations whereas the p.Ser142Phe and p.Ile360Tyrfs＊31 mutations caused the severe disease manifestation.     

Health-Related Quality of Life of Young Adults Treated with Recombinant Human Growth Hormone during Childhood

Background

Since recombinant human growth hormone (rhGH) became available in 1985, the spectrum of indications has broadened and the number of treated patients increased. However, long-term health-related quality of life (HRQoL) after childhood rhGH treatment has rarely been documented. We assessed HRQoL and its determinants in young adults treated with rhGH during childhood.

Methodology/Principal Findings

For this study, we retrospectively identified former rhGH patients in 11 centers of paediatric endocrinology, including university hospitals and private practices. We sent a questionnaire to all patients treated with rhGH for any diagnosis, who were older than 18 years, and who resided in Switzerland at time of the survey. Three hundred participants (58% of 514 eligible) returned the questionnaire. Mean age was 23 years; 56% were women; 43% had isolated growth hormone deficiency, or idiopathic short stature; 43% had associated diseases or syndromes, and 14% had growth hormone deficiency after childhood cancer. Swiss siblings of childhood cancer survivors and the German norm population served as comparison groups. HRQoL was assessed using the Short Form-36. We found that the Physical Component Summary of healthy patients with isolated growth hormone deficiency or idiopathic short stature resembled that of the control group (53.8 vs. 54.9). Patients with associated diseases or syndromes scored slightly lower (52.5), and former cancer patients scored lowest (42.6). The Mental Component Summary was similar for all groups. Lower Physical Component Summary was associated with lower educational level (coeff. -1.9). Final height was not associated with HRQoL.

Conclusions/Significance

In conclusion, HRQoL after treatment with rhGH in childhood depended mainly on the underlying indication for rhGH treatment. Patients with isolated growth hormone deficiency/idiopathic short stature or patients with associated diseases or syndromes had HRQoL comparable to peers. Patients with growth hormone deficiency after childhood cancer were at high risk for lower HRQoL. This reflects the general impaired health of this vulnerable group, which needs long-term follow-up.     

重组人类生长激素对胃癌细胞的体外增殖研究

目的：通过体外实验,研究重组人类生长激素对胃癌细胞的增殖的影响。方法：实验分为空白组,重组人类生长激素组,奥沙利铂组和重组人类生长激素＋奥沙利铂组。用不同浓度的重组人类生长激素处理SGC．7901细胞,采用MTT法和流式细胞仪检测人胃癌细胞株的细胞抑制率,细胞周期和DNA抑制率。结果：体外实验结果表明,重组人类生长激素对SGC．7901细胞株增殖没有明显的促进作用,重组人类生长激素组和空白组以及重组人类生长激素＋奥沙利铂组和奥沙利铂组之间没有统计显著性（P〉0．05）,细胞抑制率和停止生长的细胞在G0-G1期明显增加（P〈0．01）,同时重组人类生长激素＋奥沙利铂组和空白组以及奥沙利铂组在S期,细胞数依次下降,DNA抑制率依次增加。重组人类生长激素＋奥沙利铂组与奥沙利铂组相比,细胞抑制率有明显上升趋势。结论：体外实验表明,重组人类生长激素并不加快人类胃癌细胞的增殖,与抗癌药物一同使用时,有增加治疗功效的作用。     

Characterization of Recombinant Integrase of Human Immunodeficiency Virus Type 1 (Isolate Bru)

Integration of the human immunodeficiency virus type 1 (HIV-1) DNA into the human genome requires the virusencoded integrase protein. The recombinant integrase protein of HIV-1 (isolate Bru) was prepared by constructing a plasmid based on pET-15b encoding the integrase gene. Integrase of HIV-1 was purified using a bacterial expression system (Escherichia coli). The main kinetic parameters of HIV-1 integrase (K _m = (3.7 ± 0.2)·10^–10 M, k _cat = (1.2 ± 0.3)·10^–7 sec^–1) were determined using an oligonucleotide duplex constructed on the basis of the U5-terminal sequence of proviral HIV-1 DNA as the substrate. Inhibition of integrase by aurintricarbonic acid ([I]₅₀ = 6.3 ± 0.4 M) and dependence of integrase activity on Mg²⁺ and Mn²⁺ concentration were studied.     

Illness Perception and Clinical Treatment Experiences in Patients with M. Maroteaux-Lamy (Mucopolysaccharidosis Type VI) and a Turkish Migration Background in Germany

Introduction

Mucopolysaccharidosis VI (MPS VI) is an inherited lysosomal storage disease caused by a mutation of the gene for arylsulfatase B (ASB). Of the thirty-one patients registered in Germany, almost fifty percent have a Turkish migration background. MPS VI is treated by enzyme replacement therapy (ERT), which is time-consuming and expensive.

Methods

This interdisciplinary study explored the illness perceptions and clinical treatment experiences among ten MPS VI patients with a Turkish migration background in two centers for metabolic diseases (Berlin and Mainz, Germany). The clinical treatment situation was observed and semi-structured interviews were conducted with patients and health care personnel, in addition to participatory observation in four patients'' everyday environments in Berlin. The data from the interviews, patient records, and personal field notes were encoded, cross-related, and analyzed.

Results

Patients'' acknowledgement of the disease and coping strategies are influenced predominantly by the perception of their individual health status and the handling of the disease within their family. Patients'' willingness to cooperate with treatment strategies is further modified by their knowledge of the disease and the relationships with their health care providers. In this analysis, cultural factors turned out to be marginally relevant.

Conclusion

As with other chronic and debilitating diseases, effective treatment strategies have to reach beyond delivering medication. Health care providers need to strengthen the support for patients with a migration background. In this regard, they should respect the patients'' cultural and social background and their personal perception of the disease and the therapy. Yet structural and social aspects (clinical setting, family and educational background) may be more crucial here than “cultural barriers.”     

Effect of Recombinant Human Thyrotropin on the Uptake of Radioactive Iodine (123I) in Dogs with Thyroid Tumors

In humans, recombinant human thyrotropin (rhTSH) enhances radioactive iodine uptake (RAIU) in patients with differentiated thyroid cancer. No studies have been performed in veterinary medicine to optimize radioiodine treatment of thyroid cancer. The aim of this study was to evaluate the effect of rhTSH on the uptake of radioiodine-123 (¹²³I) in dogs with thyroid tumors. Nine dogs with thyroid neoplasia were included in this prospective cross-over study. The dogs were divided in 2 groups. In one group, ¹²³I was administered for a baseline RAIU determination in week 1. In week 2 (after a washout period of 2 weeks), these dogs received rhTSH (100 μg IV) 24 h before ¹²³I injection. In the other group the order of the protocol was reversed. For each scan, the dogs received 37 MBq (1 mCi) of ¹²³I intravenously (IV) and planar scintigraphy was performed after 8 and 24 h for tumor RAIU calculation. Overall, rhTSH administration caused no statistically significant change on thyroid tumor RAIU at 8 h (p = 0.89) or at 24 h (p = 0.98). A significant positive correlation was found between the effect of rhTSH on tumor 8h-RAIU and rhTSH serum concentrations at 6 h (τ = 0.68; p = 0.03), at 12 h (τ = 0.68; p = 0.03) and at 24 h (τ = 0.78; p = 0.02) after rhTSH injection. This study suggests that IV administration of 100 μg rhTSH 24 h before ¹²³I has an inconsistent effect on thyroid tumor RAIU. Further studies are necessary to determine the best protocol of rhTSH administration to optimize thyroid tumor RAIU.     

重组人生长激素治疗生长激素缺乏症的疗效观察

目的：正确评价重组人生长激素（rhGH）治疗儿童生长激素缺乏症（GHD）的疗效。方法：GHD患儿47例,给予rhGH0．1U／（kg．d）,疗程3个月,并对其疗效进行观察。结果：身高（height）由（122±17．67）cm增至（125．32±17．50）cm,生长速率（growthrate）由〈4cm／年增加到（10．40±3．74）cm／年。血清碱性磷酸酶（AKP）由（207±48．11）IU／L增至（261±45．29）IU／L。I型前胶原羧基端伸展肽（PICP）由（97．80±14．94）ng／ml增至（119．50±24．10）ng／ml值。肌酐（Cr）由（40．20±8．28）umol／L变为（36．50±8．33）umol／L值。结论：rhGH是治疗GHD有效的药物。     

The Effect of (2-chloroethyl)-trimethylammonium Chloride on the Growth of Barley

Barley (Hordeum vulgare L. C.I.666) was shown to be susceptibleto the growth retardant (2-chloroethyl)-trimethylammonium chloride(CCC). The estimation of cell number in the dwarfed third leafblade indicated that a decrease in mitotic activity had occurredin treated plants. There was also a decrease in cell size intreated plants. The dwarfing action of CCC was reversed by exogenousgibberellic acid (GA₃) but this was shown to be the result ofincreased cell elongation only. GA₃ did not promote cell divisionin healthy or CCC-treated plants. Assay of endogenous gibberellinsshowed a significant reduction in the level of a substance correspondingto GA₃ in CCC-treated plants. It is suggested that CCC-induceddwarfing of barley is largely the result of a reduction in meristematicactivity. This may be related to an effect on gibberellin biosynthesisbut is not reversed by the application of exogenous GA ₃.     

胰岛素泵强化治疗对初诊2型糖尿病的疗效观察

目的:观察胰岛素泵强化治疗对合并肥胖和高脂血症的初诊2型糖尿病(T2DM)患者的疗效。方法:采用胰岛素泵持续皮下输注(CS‖)超短效门冬胰岛素(诺和锐)强化治疗15天(15d)54例合并肥胖和高脂血症的新诊断T2DM患者,分别在强化治疗15d结束时和继治疗90d时观察患者的血糖、糖化血红蛋白、血脂、体重指数和反应胰岛β细胞功能的胰岛素和C肽指标改变情况。结果:在强化治疗15d撤泵时和治疗90d后患者血糖明显下降;而糖化血红蛋白、血脂、体重指数、胰腺β细胞功能在治疗90d时改善明显。结论:早期胰岛素泵强化治疗新诊断的2型糖尿病(T2DM)患者,不仅可使其血糖尽早达标,而且还可明显降低血脂和体重指数、糖化血红蛋白,并使部分患者胰岛β细胞功能恢复,一段时期内脱离药物治疗,提高生活质量。     

心理干预对中青年肺结核患者治疗效果的影响

目的:探讨心理干预对中青年肺结核患者治疗效果的影响.方法:干预组60例,在常规抗结核药物治疗的基础上进行心理干预,对照组55例单纯进行常规抗结核药物治疗,在疗程结束时对两组患者进行依从性调查,同时进行痰涂片及胸部X线检查,比较两组疗效,以观察心理干预在抗结核治疗期间的作用.结果:干预组在依从性、痰菌阴转率及病灶吸收率方面均高于对照组,差异有统计学意义(P<0.05).结论:心理干预可以提高患者抗结核治疗的依从性,提高疗效.     

Human T-Lymphotropic Virus Type 2 (HTLV-2) Provirus in Circulating Cells of the Monocyte/Macrophage Lineage in Patients Dually Infected with Human Immunodeficiency Virus Type 1 and HTLV-2 and Having Predominantly Sensory Polyneuropathy

We investigated the presence of human T-lymphotropic virus type 2 (HTLV-2) DNA in the peripheral blood mononuclear cell subsets obtained from 18 patients coinfected with human immunodeficiency virus type 1 and HTLV-2, 6 of whom also had predominantly sensory polyneuropathy (PSP). HTLV-2 DNA and RNA were found in CD8- and CD19-positive cells, and, for patients with PSP, in CD14-positive cells as well. Furthermore, the patients with PSP had higher proviral loads than those without PSP.     

Mucopolysaccharidosis type II (Hunter syndrome): mutation "hot spots" in the iduronate-2-sulfatase gene.

Mucopolysaccharidosis type II (MPS II, Hunter syndrome) is an X-chromosomal storage disorder due to deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). We have identified IDS mutations in a total of 31 families/patients with MPS II, of which 20 are novel and unique and a further 1 is novel but has been found in 3 unrelated patients. One of the mutations detected is of special interest as an AG-->G substitution in an intron, far apart from the coding region, is deleterious by creating a new 5''-splice-donor site that results in the inclusion of a 78-bp intronic sequence. While the distribution of gene rearrangements (deletions, insertions, and duplications) of <20 bp seems to be random over the IDS gene, the analysis of a total of 101 point mutations lying within the coding region shows that they tend to be more frequent in exons III, VIII, and IX. Forty-seven percent of the point mutations are at CpG dinucleotides, of which G:C-to-A:T transitions constitute nearly 80%. Almost all recurrent point mutations involve CpG sites. Analysis of a collective of 50 families studied in our laboratory, to date, revealed that mutations occur more frequently in male meioses (estimated male-to-female ratio between 3.76 and 6.3).     

The Distribution of High-Risk Human Papillomaviruses Is Different in Young and Old Patients with Cervical Cancer

Despite numerous human papillomavirus (HPV) frequency studies in women with cervical cancer (CC), little is known of HPV frequency trends according to patient age. In this work, we compare the mean age and frequency distribution by age of CC patients positive for different HPVs. This study included 462 CC patients. HPVs were detected by PCR and typed using DNA sequencing. A total of 456 patients (98.7%) were positive for HPV: 418 (90.5%) had single and 38 (8.2%) had double HPV infections. HPV16 (46.5%), HPV18 (10.4%), HPV45 (6.7%), and HPV31 (4.1%) were the most frequent viral types in single-infected patients. The mean ages of single-infected patients with HPV16 (49.2±13.3), HPV18 (47.9±12.2), HPV45 (47.9±11.7), or HPV39 (42.6±8.9) were significantly lower than the mean ages of patients singly (53.9±12.7; p<0.001, t-test) or doubly (55.4±12.7; p<0.05, t-test) infected with the remaining HPVs. Three different trends were identified: one for HPV16, another for HPVs18/45/39, and a third for the rest of HPVs. The frequency trend of HPV16 shows two peaks. The first (63.2%) was found in the youngest women (≤35 years), followed by a decreasing trend until the age of 55–60 years (31.1%). The second peak arose at 61–65 years (52.5%), followed by a decreasing trend. The trend for HPVs18/45/39 declined from the youngest (19.3%) to the oldest (>70 years; 12.8%) women. In contrast, the trend for the remaining HPVs increased from the youngest (15.8%) to the oldest (46.2%) women. Unlike other life-style factors, low-risk sexual behavior was associated with late onset of CC independent of low-oncogenic HPV types (p<0.05, Wald chi-square statistic). The data indicate that most CCs in young women depend on the presence of high-oncogenic HPVs. In contrast, almost half of CCs in older patients had low-oncogenic HPVs, suggesting they could depend on the presence of other factors.     

Post-translational Modifications of Recombinant Human Lysyl Oxidase-like 2 (rhLOXL2) Secreted from Drosophila S2 Cells

Human lysyl oxidase-like 2 (hLOXL2) is highly up-regulated in metastatic breast cancer cells and tissues and induces epithelial-to-mesenchymal transition, the first step of metastasis/invasion. hloxl2 encodes four N-terminal scavenger receptor cysteine-rich domains and the highly conserved C-terminal lysyl oxidase (LOX) catalytic domain. Here, we assessed the extent of the post-translational modifications of hLOXL2 using truncated recombinant proteins produced in Drosophila S2 cells. The recombinant proteins are soluble, in contrast to LOX, which is consistently reported to require 2–6 m urea for solubilization. The recombinant proteins also show activity in tropoelastin oxidation. After phenylhydrazine derivatization and trypsin digestion, we used mass spectrometry to identify peptides containing the derivatized lysine tyrosylquinone cross-link at Lys-653 and Tyr-689, as well as N-linked glycans at Asn-455 and Asn-644. Disruption of N-glycosylation by site-directed mutagenesis or tunicamycin treatment completely inhibited secretion so that only small quantities of inclusion bodies were detected. The N-glycosylation site at Asn-644 in the LOX catalytic domain is not conserved in human LOX (hLOX), although the LOX catalytic domain of hLOX shares ∼50% identity and ∼70% homology with hLOXL2. The catalytic domain of hLOX was not secreted from S2 cells using the same expression system. These results suggest that the N-glycan at Asn-644 of hLOXL2 enhances the solubility and stability of the LOX catalytic domain.