[Clinical features of 19 cases of lower respiratory papillomatosis].

Objective: To explore the clinical characteristics and prognosis of lower respiratory papilloma(LRP)and therefore to improve clinical diagnosis and treatment. Methods: We performed a retrospective analysis of patients who were diagnosed with LRP in our department from October 2008 to October 2020. Results: Nineteen patients were enrolled and 12 were male and 7 were female. The average age of the 7 adult patients was (41.3±17.5)years and that of the 12 pediatric patients was (5.5±3.5)years. Ten (83.3%)of the pediatric patients showed disease onset at an age of less than 5 years. The main symptoms were cough and sputum production (13/19), dyspnea (15/19), hoarseness (10/19) and signs of stridor or wheezing (7/19). Chest CT examination was performed in 9 patients, which showed nodules or masses (9/9), cystic thin-walled cavity (4/9), obstructive pneumonia (2/9), atelectasis (2/9), and spicule sign (1/9). The upper respiratory tract was affected in all the pediatric patients (12/12) and 3/7 of the adult patients. Eighteen cases (18/19) were diagnosed by bronchoscopy, 1 (1/9) by thoracoscopy. Eighteen cases (18/19) showed mulberry-like and papillary lesions under bronchoscopy. All the cases were histologically confirmed as squamous cell papilloma, with 17 cases(17/19)showing tissue HPV6/11(+), 2 negative (2/19). The positive rate of HPV6 was 36.8%, ant that of HPV11 was 21.1%, while the double positive rate of HPV6/11 was 31.6%, and HPV16/18 were negative in all the 19 cases. Isolated respiratory papillomatosis was found in 4 cases (4/19), and multiple papillomatosis in 15 cases (15/19). Seventeen cases (17/19) underwent endoscopic interventional therapy, and the result showed that 15 cases relapsed, and 2 cases had no recurrence. One patient was treated with thoracoscopic lobectomy, and died 4 months after surgery. One patient gave up treatment. Conclusions: LRP is a rare clinical disease with a chronic course, and isolated LRP is even rarer. Young patients often suffer from upper respiratory tract involvement, and the main symptoms are cough, sputum production, dyspnea and hoarseness. CT scanning showed nodules and masses, cystic thin-walled cavities or signs of airway obstruction. Bronchoscopy often demonstrates papillary lesions. The diagnosis depends on pathology, with squamous cell papilloma being the most common, and most tests are positive for HPV6/11. It is suggested that the incidence is associated with low-risk HPV infection. Endoscopic resection is the main treatment, which is prone to relapse. The treatment should take into account the pathological changes of upper respiratory tract, and the etiological treatment of HPV should be stressed.

Extrication process of chlorogenic acid in Crofton weed and antibacterial mechanism of chlorogenic acid on Escherichia coli.

Crofton weed is a perennial herb and a biological intruding species. The present study firstly used the orthogonal test to compare the differences in extraction of chlorogenic acid in leaves and stems of Crofton weed by using three kinds of solvents, namely water, ethanol and ethyl acetate. The best effect was found by using ethonolic extraction of chlorogenic acid in Crofton weed. Further, by choosing Escherichia coli as test object, in-vitro antibacterial test was conducted to study the antimicrobial activities of chlorogenic acid by testing a series of indexes before and after the interaction between chlorogenic acid and Escherichia coli, to clarify the antibacterial mechanism of chlorogenic acid on Escherichia coli. Finally, by comparing the antibacterial activities of isochlorogenic acid A on Escherichia coli, it was concluded that both chlorogenic acid and isochlorogenic acid A showed antibacterial activities against Escherichia coli, wherein chlorogenic acid had a better antibacterial effect on Escherichia coli than isochlorogenic acid A.

Sox9 is required for cartilage formation.

Chondrogenesis results in the formation of cartilages, initial skeletal elements that can serve as templates for endochondral bone formation. Cartilage formation begins with the condensation of mesenchyme cells followed by their differentiation into chondrocytes. Although much is known about the terminal differentiation products that are expressed by chondrocytes, little is known about the factors that specify the chondrocyte lineage. SOX9 is a high-mobility-group (HMG) domain transcription factor that is expressed in chondrocytes and other tissues. In humans, SOX9 haploinsufficiency results in campomelic dysplasia, a lethal skeletal malformation syndrome, and XY sex reversal. During embryogenesis, Sox9 is expressed in all cartilage primordia and cartilages, coincident with the expression of the collagen alpha1(II) gene (Col2a1) . Sox9 is also expressed in other tissues, including the central nervous and urogenital systems. Sox9 binds to essential sequences in the Col2a1 and collagen alpha2(XI) gene (Col11a2) chondrocyte-specific enhancers and can activate these enhancers in non-chondrocytic cells. Here, Sox9 is identified as a regulator of the chondrocyte lineage. In mouse chimaeras, Sox9-/- cells are excluded from all cartilages but are present as a juxtaposed mesenchyme that does not express the chondrocyte-specific markers Col2a1, Col9a2, Col11a2 and Agc. This exclusion occurred cell autonomously at the condensing mesenchyme stage of chondrogenesis. Moreover, no cartilage developed in teratomas derived from Sox9-/- embryonic stem (ES) cells. Our results identify Sox9 as the first transcription factor that is essential for chondrocyte differentiation and cartilage formation.

Effects of antinutritional factors on plasma lipoprotein levels in Japanese flounder Paralichthys olivaceus.

This study examined the effects of four types of antinutritional factor (phytic acid, stachyose, soy saponins and soy isoflavones) on lipoprotein levels in plasma of Japanese flounder Paralichthys olivaceus. A basal diet was prepared with fish meal as primary protein source, the other diets were supplemented with 0·2, 0·4 or 0·8% phytic acid, 0·4, 0·8 or 1·5% stachyose, 0·1, 0·35 or 0·7% soy saponins and 0·10, 0·35 or 0·70% soy isoflavones, by dry mass, in place of white flour in the basal diet. Total cholesterol (TC) and triglyceride (TG) levels in plasma of P. olivaceus were not affected by phytic acid or stachyose. In general, addition of 0·2-0·8% phytic acid or 0·4-1·5% stachyose decreased plasma high-density lipoprotein cholesterol (HDL-C) levels, increased plasma low-density lipoprotein cholesterol (LDL-C) levels, thereby increasing the LDL-C:HDL-C ratio. By contrast, supplementation with 0·35-0·7% soy saponins generally depressed plasma TC levels and the LDL-C:HDL-C ratio. Supplementation with 0·35-0·7% soy isoflavones, however, increased plasma TC and TG levels. These results indicate that soy saponins may be partly responsible for the cholesterol-lowering effects of soybean meal.

The transcription factors L-Sox5 and Sox6 are essential for cartilage formation.

L-Sox5 and Sox6 are highly identical Sry-related transcription factors coexpressed in cartilage. Whereas Sox5 and Sox6 single null mice are born with mild skeletal abnormalities, Sox5; Sox6 double null fetuses die with a severe, generalized chondrodysplasia. In these double mutants, chondroblasts poorly differentiate. They express the genes for all essential cartilage extracellular matrix components at low or undetectable levels and initiate proliferation after a long delay. All cartilages are thus extracellular matrix deficient and remain rudimentary. While chondroblasts in the center of cartilages ultimately activate prehypertrophic chondrocyte markers, epiphyseal chondroblasts ectopically activate hypertrophic chondrocyte markers. Thick intramembranous bone collars develop, but the formation of cartilage growth plates and endochondral bones is disrupted. L-Sox5 and Sox6 are thus redundant, potent enhancers of chondroblast functions, thereby essential for endochondral skeleton formation.

Epithelial neutrophil-activating peptide-78 recruits neutrophils into pleural effusion.

The aim of this study was to investigate the presence of epithelial neutrophil-activating peptide (ENA)-78 in pleural effusions, as well as the chemoattractant activity of pleural ENA-78 on neutrophils. Pleural effusion and serum samples were collected from 75 patients who presented to the respiratory institute (19 with malignant pleural effusion, 21 with tuberculous pleural effusion, 18 with infectious pleural effusion and 17 with transudative pleural effusion). The concentrations of ENA-78, myeloperoxidase and neutrophil elastase were determined, and the chemoattractant activity of ENA-78 for neutrophils both in vitro and in vivo was also observed. The concentrations of ENA-78, myeloperoxidase and neutrophil elastase in infectious pleural effusion were significantly higher than those in malignant, tuberculous and transudative groups, respectively (all p<0.01). Infectious pleural fluid was chemotactic for neutrophils in vitro and anti-ENA-78 antibody could partly inhibit these chemotactic effects. Intrapleural administration of ENA-78 produced a marked progressive influx of neutrophils into pleural space. Compared with noninfectious pleural effusion, ENA-78 appeared to be increased in infectious pleural effusion. Our data suggested that ENA-78 was able to induce neutrophil infiltration into pleural space and might be responsible for pleural neutrophil degranulation.

Targeted disruption of the transition protein 2 gene affects sperm chromatin structure and reduces fertility in mice.

During mammalian spermiogenesis, major restructuring of chromatin takes place. In the mouse, the histones are replaced by the transition proteins, TP1 and TP2, which are in turn replaced by the protamines, P1 and P2. To investigate the role of TP2, we generated mice with a targeted deletion of its gene, Tnp2. Spermatogenesis in Tnp2 null mice was almost normal, with testis weights and epididymal sperm counts being unaffected. The only abnormality in testicular histology was a slight increase of sperm retention in stage IX to XI tubules. Epididymal sperm from Tnp2-null mice showed an increase in abnormal tail, but not head, morphology. The mice were fertile but produced small litters. In step 12 to 16 spermatid nuclei from Tnp2-null mice, there was normal displacement of histones, a compensatory translationally regulated increase in TP1 levels, and elevated levels of precursor and partially processed forms of P2. Electron microscopy revealed abnormal focal condensations of chromatin in step 11 to 13 spermatids and progressive chromatin condensation in later spermatids, but condensation was still incomplete in epididymal sperm. Compared to that of the wild type, the sperm chromatin of these mutants was more accessible to intercalating dyes and more susceptible to acid denaturation, which is believed to indicate DNA strand breaks. We conclude that TP2 is not a critical factor for shaping of the sperm nucleus, histone displacement, initiation of chromatin condensation, binding of protamines to DNA, or fertility but that it is necessary for maintaining the normal processing of P2 and, consequently, the completion of chromatin condensation.

Two case reports of Castleman disease with pulmonary involvement.

Castleman disease (CD) is a rare reactive lymphoproliferative disorder, first identified in 1954. We recently had the opportunity to analyse the characteristics of two variations of CD with pulmonary involvement. Case 1 had localised retroperitoneal hyaline vascular type CD, while Case 2 was diagnosed as multicentric plasma cell type CD. Both patients had pulmonary symptoms and signs, including cough, dyspnoea, hypoxaemia and ventilatory dysfunction; however, they had different physiological manifestations of their pulmonary abnormalities.

Efficacy of intensified chemotherapy with hematopoietic progenitors in small-cell lung cancer: A meta-analysis of the published literature.

OBJECTIVE: It remains controversial whether intensified chemotherapy with hematopoietic progenitors (ICHP) is effective for small-cell lung cancer. This meta-analysis was performed to evaluate the efficacy and safety of ICHP in patients with small-cell lung cancer. METHODS: MEDLINE and EMBASE databases were searched for English-language studies published through October 12, 2008. Randomized phase II and III clinical trials comparing ICHP with control therapy. Response rates, overall survival, and toxicity were analyzed. RESULTS: Five assessable trials were identified including 641 patients. No significant increase in the odds ratio for response was attributable to ICHP (odds ratio, 1.29; 95% confidence interval, 0.87-1.93; p=0.206). No statistically significant increase in overall survival was found when ICHP were compared to control regimens (hazard ratio, 0.94; 95% confidence interval, 0.80-1.10; p=0.432). The toxicity of ICHP was significantly higher for hematologic toxicity, including hemoglobin nadir and platelet nadir. CONCLUSIONS: ICHP was not superior to control chemotherapy in terms of both objective response and overall survival, and was related to more significant hemoglobin nadir and platelet nadir.

An insertional mutation in the BTF3 transcription factor gene leads to an early postimplantation lethality in mice.

The gene that encodes the general transcription factor known as BTF3 was disrupted in mouse embryonic stem cells in a random mutagenesis screen for developmental genes with the ROSA beta-geo (Friedrich and Soriano, 1991) retroviral gene trap vector. The BTF3 mutation was transmitted through the germline of chimaeric mice. While the endogenous BTF3 gene is ubiquitously expressed, the expression pattern of the beta-galactosidase reporter gene present in the gene trap vector in BTF3 heterozygotes was restricted. Mice homozygous for the mutant allele died soon after implantation, around embryonic day 6.5. Thus, BTF3 is essential for postimplantation development. The isolation of the BTF3 sequences in this ROSA beta-geo insertion was facilitated by a relatively simple single lacZ primer reverse transcription PCR strategy.

Expression of the mouse cerberus-related gene, Cerr1, suggests a role in anterior neural induction and somitogenesis.

The Xenopus cerberus gene encodes a secreted factor that is expressed in the anterior endomesoderm of gastrula stage embryos and can induce the formation of ectopic heads when its mRNA is injected into Xenopus embryos [Bouwmeester, T., Kim, S., Lu, B. & De Robertis, E. M. (1996) Nature (London) 382, 595-601]. Here we describe the existence of a cerberus-related gene, Cerr1, in the mouse. Cerr1 encodes a putative secreted protein that is 48% identical to cerberus over a 110-amino acid region. Analysis of a mouse interspecific backcross panel demonstrated that Cerr1 mapped to the central portion of mouse chromosome 4. In early gastrula stage mouse embryos, Cerr1 is expressed in the anterior visceral endoderm and in the anterior definitive endoderm. In somite stage embryos, Cerr1 expression is restricted to the most recently formed somites and in the anterior presomitic mesoderm. Germ layer explant recombination assays demonstrated that Cerr1-expressing somitic-presomitic mesoderm, but not older Cerr1-nonexpressing somitic mesoderm, was able to mimic the anterior neuralizing ability of anterior mesendoderm and maintain Otx2 expression in competent ectoderm. In most Lim1-/- headless embryos, Cerr1 expression in the anterior endoderm was weak or absent. These results suggest that Cerr1 may play a role in anterior neural induction and somite formation during mouse development.

Sequence and genomic organization of the mouse Lim1 gene.

The sequence and genomic organization of the mouse Lim1 gene were determined. The mouse Lim1 gene has five coding exons. The Lim1 transcription initiation start site was determined by 5' RACE. indicating that the first exon encodes the translation initiation codon and a 1360-bp 5' untranslated region. Sequence analysis of the 450-bp upstream of the transcription start site revealed the presence of a CATTAA motif at -32 bp and a CAATT box located in reverse orientation at -68 bp. HNF3 beta and Pbx1 binding sites were also identified. Like most LIM domain encoding genes, the LIM domains of Lim1 are each encoded on separate and adjacent exons. Knowledge of the sequence and structure of the mouse Lim1 gene provides important information for the genetic manipulation of the Lim1 locus.

Overlapping roles and asymmetrical cross-regulation of the USF proteins in mice.

USF1 and USF2 are ubiquitously expressed transcription factors implicated as antagonists of the c-Myc protooncoprotein in the control of cellular proliferation. To determine the biological role of the USF proteins, mutant mice were generated by homologous recombination in embryonic stem cells. USF1-null mice were viable and fertile, with only slight behavioral abnormalities. However, these mice contained elevated levels of USF2, which may compensate for the absence of USF1. In contrast, USF2-null mice contained reduced levels of USF1 and displayed an obvious growth defect: they were 20-40% smaller at birth than their wild-type or heterozygous littermates and maintained a smaller size with proportionate features throughout postnatal development. Some of the USF-deficient mice, especially among the females, were prone to spontaneous epileptic seizures, suggesting that USF is important in normal brain function. Among the double mutants, an embryonic lethal phenotype was observed for mice that were homozygous for the Usf2 mutation and either heterozygous or homozygous for the Usf1 mutation, demonstrating that the USF proteins are essential in embryonic development.

Restricted beta-galactosidase expression of a hygromycin-lacZ gene targeted to the beta-actin locus and embryonic lethality of beta-actin mutant mice.

beta-actin is a cytoskeletal protein that is ubiquitously expressed. To exploit the regulation the beta-actin gene, a promoterless hygromycin-lacZ fusion gene with a splice acceptor was introduced into the first intron of the beta-actin locus by homologous recombination in mouse embryonic stem (ES) cells. The targeted ES cells were hygromycin resistant and expressed beta-galactosidase (beta-gal) activity. However, no beta-gal activity was detected in heterozygous embryos. In adult heterozygotes, beta-gal activity was detected only in testes. RT-PCR analysis demonstrated the presence of both beta-actin exon 1-hygromycin- and exon l-exon 2-containing transcripts in homozygous mutant embryos. LacZ-containing transcripts were detected in adult heterozygous tests and, surprisingly, in homozygous mutant embryos. These results demonstrate that the integration of the hygromycin-lacZ gene into the first intron of the beta-actin locus was not productive for the ubiquitous expression of beta-gal activity. Because this integration mimics certain types of gene trap events, it suggests that caution should be used when interpreting beta-gal expression patterns in genetic screens using gene trap strategies. In addition, mice homozygous for the beta-actin mutation developed normally up to embryonic day 8.5 (E8.5) but became growth retarded at E9.5 and subsequently died. The RT-PCR data indicate that this targeted mutation is a hypomorphic allele of beta-actin.

Clinical application of therapeutic hemapheresis.

Hemapheresis has been used in a variety of clinical states, primarily for its ability to remove an offending component, likely be either plasma or cellular elements. We have reviewed 43 cases of therapeutic hemapheresis over the past two years at Veterans General Hospital. There were 20 cases of plasmapheresis, 7 cases of leukapheresis and 16 cases of thrombocytapheresis. Most of them had satisfactory and desirable effects except a few cases in therapeutic plasmapheresis. Among 20 cases of therapeutic plasmapheresis, all but 2 cases of systemic lupus erythrematosus had transient clinical improvement. Average decrease of leukocyte count in 11 procedures of leukemic patients was 37%. As for the myeloproliferative disorders with thrombocytosis, the average drop of platelet count was 41%. The side effects of therapeutic hemapheresis were not infrequent. There were 56 recorded side effects with one sudden death among 94 procedures. Hemapheresis is useful in certain clinical conditions but more judicious application should be considered.

Soluble CD86 protein in serum samples of patients with asthma.

BACKGROUND: Previous studies have reported that soluble (s) CD86 is involved in the initiation of the immune response. A study was undertaken to investigate the concentrations of sCD86 in serum samples from patients with bronchial asthma and to determine the cell origin of sCD86. METHODS: Serum sCD86 concentrations were measured in 52 asthmatic subjects and 25 non-atopic normal volunteers using an enzyme linked immunosorbent assay, and the relationship of serum sCD86 concentrations to asthma severity and to total and differential white cell counts was analysed. Each type of white blood cell was purified and cultured in vitro to determine the cell origin of serum sCD86. RESULTS: Serum samples from patients with an acute asthma exacerbation had much higher levels of sCD86 (585.4 (20.5) IU/ml) than those from stable asthmatics (479.6 (15.7) IU/ml, p<0.001) and healthy individuals (435.1 (13.8) IU/ml, p<0.001), and there was no difference between the latter two groups (p = 0.079). In asthmatic subjects the serum sCD86 level was inversely correlated with airway responsiveness, forced expiratory volume in 1 second, and with arterial carbon dioxide tension. In addition, the serum sCD86 level was positively correlated with numbers of lymphocytes, eosinophils, monocytes, but not neutrophils. The in vitro experiments indicated that sCD86 was produced by monocytes. CONCLUSIONS: The serum sCD86 protein level was significantly increased in asthmatic subjects during an exacerbation and correlated with the severity of asthma. sCD86 is most probably derived from monocytes in the peripheral blood.

Successful management of alloimmunized patients by transfusing HLA compatible and crossmatched platelets.

An average of three (range 0-22) perfect matches for 65 patients could be found in 1490 filed HLA-typed donors. It could increase to 8.7 (0-45) if strong cross-reactive antigens were added. Three methods of platelet crossmatching, i.e, lymphocytotoxicity test (LCT), platelet suspension immunofluorescence test (PSIFT) and solid phase red cell adherence test (SPRCA), were used to search for compatible donors. Forty episodes of apheresis platelet transfusion were evaluated in seven patients. Using one-hour posttransfusional corrected count of increment (CCI), greater than 7500 microliters as a guideline to assess the reliability of these three methods, we found that the efficiency of PSIFT, LCT and SPRCA were 67.5%, 97.5% and 97.5% respectively. Retrospectively, we did the HLA-A, B typing of those matched donors. There was no significant difference in CCI among different HLA match grades. The concordance of LCT and SPRCA was 100%, indicating that all the antibodies were of HLA specificity and there was no platelet specific antibody. We concluded that HLA donor-file and platelet crossmatching technique could be used together in selecting suitable donors for alloimmunized patients.

Haploinsufficiency of Sox9 results in defective cartilage primordia and premature skeletal mineralization.

In humans, SOX9 heterozygous mutations cause the severe skeletal dysmorphology syndrome campomelic dysplasia. Except for clinical descriptions, little is known about the pathogenesis of this disease. We have generated heterozygous Sox9 mutant mice that phenocopy most of the skeletal abnormalities of this syndrome. The Sox9(+/-) mice died perinatally with cleft palate, as well as hypoplasia and bending of many skeletal structures derived from cartilage precursors. In embryonic day (E)14.5 heterozygous embryos, bending of radius, ulna, and tibia cartilages was already prominent. In E12.5 heterozygotes, all skeletal elements visualized by using Alcian blue were smaller. In addition, the overall levels of Col2a1 RNA at E10.5 and E12.5 were lower than in wild-type embryos. We propose that the skeletal abnormalities observed at later embryonic stages were caused by delayed or defective precartilaginous condensations. Furthermore, in E18.5 embryos and in newborn heterozygotes, premature mineralization occurred in many bones, including vertebrae and some craniofacial bones. Because Sox9 is not expressed in the mineralized portion of the growth plate, this premature mineralization is very likely the consequence of allele insufficiency existing in cells of the growth plate that express Sox9. Because the hypertrophic zone of the heterozygous Sox9 mutants was larger than that of wild-type mice, we propose that Sox9 also has a role in regulating the transition to hypertrophic chondrocytes in the growth plate. Despite the severe hypoplasia of cartilages, the overall organization and cellular composition of the growth plate were otherwise normal. Our results suggest the hypothesis that two critical steps of the chondrocyte differentiation pathway are sensitive to Sox9 dosage. First, an early step presumably at the stage of mesenchymal condensation of cartilage primordia, and second, a later step preceding the transition of chondrocytes into hypertrophic chondrocytes.

Abnormal spermatogenesis and reduced fertility in transition nuclear protein 1-deficient mice.

Transition nuclear proteins (TPs), the major proteins found in chromatin of condensing spermatids, are believed to be important for histone displacement and chromatin condensation during mammalian spermatogenesis. We generated mice lacking the major TP, TP1, by targeted deletion of the Tnp1 gene in mouse embryonic stem cells. Surprisingly, testis weights and sperm production were normal in the mutant mice, and only subtle abnormalities were observed in sperm morphology. Electron microscopy revealed large rod-like structures in the chromatin of mutant step 13 spermatids, in contrast to the fine chromatin fibrils observed in wild type. Steps 12-13 spermatid nuclei from the testis of Tnp1-null mice contained, in place of TP1, elevated levels of TP2 and some protamine 2 (P2) precursor. Most of the precursor was processed to mature P2, but high levels of incompletely processed forms remained in epididymal spermatozoa. Sperm motility was reduced severely, and approximately 60% of Tnp1-null males were infertile. We concluded that TP1 is not essential for histone displacement or chromatin condensation. The absence of TP1 may partially be compensated for by TP2 and P2 precursor, but this dysregulation of nucleoprotein replacement results in an abnormal pattern of chromatin condensation and in reduced fertility.