The Suppression Effects of Thalidomide on Human Lung Fibroblasts: Cell Proliferation, Vascular Endothelial Growth Factor Release, and Collagen Production

Background

Expression of transforming growth factor (TGF)-β₁ and increases in angiogenesis and deposition of extracellular matrix are the key features of tracheal granulation formation. The aim of this study was to investigate the potential role of thalidomide in preventing granulation tissue formation from the aspect of cellular effects in vitro, including fibroblast proliferation, vascular endothelial growth factor (VEGF) release, and collagen production.

Methods

Human lung fibroblasts were obtained from bronchus and cultured. The effects of thalidomide on cell proliferation, migration, TGF-β₁-induced VEGF, and signal pathway were investigated.

Results

Thalidomide (20 μM) not only inhibited cell proliferation after 24 h [fold increase of cell number, 0.85 ± 0.09 vs. 1.47 ± 0.14 (treatment vs. control group); P < 0.01] and 48 h of incubation (0.85 ± 0.10 vs. 1.97 ± 0.12; P < 0.001), it also inhibited cell migration and slowed wound closure at 24 h (P < 0.001). Thalidomide significantly attenuated TGF-β₁-induced VEGF expression at both the mRNA and protein levels. Incubation of thalidomide with cells stimulated with TGF-β₁ significantly inhibited their production of collagen. Thalidomide inhibited Smad3, STAT3, and subsequent p44/42 kinase phosphorylation.

Conclusion

Thalidomide may inhibit human fibroblast proliferation and it is worthy of further in vivo investigation.     

Childhood tuberculosis presenting as an anterior chest wall abscess.

Chest wall abscess is a rare manifestation of childhood tuberculosis. We report a case of a tuberculous chest wall abscess in a 4-year-old healthy girl who had received Bacillus Calmette-Guerin (BCG) vaccination at birth. She developed a localized anterior chest wall mass, which was initially mistaken for enchondroma on the chest radiograph. Pathologic examination of the biopsy specimen revealed chronic granulomatous inflammation and positive acid-fast staining, which confirmed the diagnosis of chest wall tuberculosis infection. She received a 12-month course of anti-tuberculous treatment and was perfectly well 1 year later. The chest wall lesion resolved without the need for surgery. In conclusion, tuberculosis should be excluded in children with undiagnosed chest wall lesions, especially in endemic areas, even if they have been vaccinated with BCG. Adequate anti-tuberculosis treatment can result in a complete recovery.     

Varicella arthritis diagnosed by polymerase chain reaction.

We report a 2-year-old girl who developed acute arthritis of the left knee 4 days after the onset of a typical varicella infection. She was first thought to have pyogenic arthritis caused by Staphylococcus aureus. Accordingly, oxacillin was administered upon hospitalization. On the third day after hospitalization, bacterial cultures of the synovial fluid and blood showed no growth and oxacillin was discontinued. Although a viral culture of the synovial fluid for varicella-zoster virus (VZV) was negative, varicella DNA was identified by means of polymerase chain reaction (PCR) with VZV-specific primers. The patient recovered spontaneously. To differentiate this condition from septic arthritis is important. PCR is a sensitive technique that can demonstrate the presence of VZV DNA in synovial fluid, even if viral cultures are negative.     

Further characterization of a potent immunogen and the chromosomal gene encoding it in the Lyme disease agent, Borrelia burgdorferi.

Further characterization of a previously reported 83-kDa antigen of Borrelia burgdorferi and the gene encoding it is reported. The DNA sequence of the gene and the amino acid sequence of the protein were determined. On the basis of the amino acid content, the actual size of the antigen was determined to be 79.8 kDa, rather than 83 kDa as previously reported. The expression of the antigen in several representative North American and European B. burgdorferi isolates was demonstrated. The conservation of the gene within the species was demonstrated by DNA hybridization of a labeled gene probe to several North American and European B. burgdorferi isolates. Of other Borrelia spp. assayed (B. hermsii, B. coriaceae, B. duttonii, and B. anserina), only B. anserina, a poultry pathogen, hybridized to the gene probe and expressed the 79.8-kDa antigen.     

Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization.

Genomes of several Borrelia burgdorferi isolates from North America and Europe were characterized by restriction endonuclease analysis and DNA hybridization using labeled B. burgdorferi whole-cell DNA (strain ATCC 35210). Several different restriction and homology patterns were observed among these isolates, indicating genotypic heterogeneity within this genus and species. It was concluded from this study that restriction endonuclease analysis of B. burgdorferi whole-cell DNA may be a reliable and accurate method for identifying strains or genotypes of the Lyme disease agent.     

An in vitro approach to the kinetics of insulin antibodies in diabetic patients

The purpose of this study is to evaluate the binding behavior of plasma insulin antibodies (Iab) in diabetic patients, by the following parameters: Incubation time, temperature, buffer pH, and Iab titer. We investigated the plasma insulin binding patterns of 12 insulin-treated diabetic patients, and they were separated as higher titer group (Iab = 51.9 +/- 7.28, n = 6) and lower titer group (Iab = 14.88 +/- 4.75, n = 6). The procedure was: (1) plasma samples were deinsulinized by 0.12N HCL, dextran coated charcoal suspension and 0.12N NaOH. (2) I-125 monoiodoinsulin was used to prevent artifacts resulting from variability in ligand binding due to excessive iodination, (3) separation of free and bound insulin was accomplished by rapid precipitation of hormone-antibody complex with polyethylene glycol, and (4) decanting the supernatants and counting the pellets in the automatic gamma counter. The data were obtained as the condition of incubation time, temperature, buffer pH, and Iab titer. The results obtained by the Scatchard analysis indicated that high temperature (39 degrees C vs 37 degrees C) in vitro would increase the free insulin levels and decrease the low affinity binding capacity (Q2-Q1) of Iab in patients with high titer of Iab (greater than 40%), whereas this phenomenon is not observed in patients with low affinity binding sites of Iab in patients with low titers of insulin antibodies (less than 20%).     

A randomized phase II study of vinorelbine plus gemcitabine with/without cisplatin against inoperable non-small-cell lung cancer previously untreated

Phase II studies have suggested that vinorelbine (V) plus gemcitabine (G) treatment has a similar response rate and better toxicity profile than cisplatin-based combination chemotherapy in non-small-cell lung cancer (NSCLC). Our aim was to evaluate whether or not the addition of cisplatin (P) to a VG regimen increases the efficacy or toxicities in chemo-naive inoperable NSCLC patients. From April 2002 to October 2003, 86 patients were enrolled. The treatment dose was V 20 mg/m2 plus G 800 mg/m2 intravenous infusion (i.v.) on days 1, 8 and 15, with/without P 60 mg/m2 i.v. on day 15, every 4 weeks. The efficacy and toxicity of the treatment were recorded. In all, 125 cycles of VG and 178 cycles of VGP were given to the patients in the VG and VGP arms, respectively (P = 0.001). The median cycle of treatment was three in the VG arm and five in the VGP arm. There were 10 partial responses (overall 23.3%) in the VG arm and 1 complete response and 19 partial responses (overall 46.5%) in the VGP arm (P = 0.022). Neutropenia, nausea, vomiting, and peripheral neuropathy were more common in the VGP arm (P = 0.023, 0.002, 0.025, 0.001, respectively). The Lung Cancer Symptom Scale showed no difference between the VG and VGP arms after two cycles of treatment or when the patient went off study. We concluded that the addition of P to VG treatment did increase both the tumor response rate and the toxicities. However, the toxicities were tolerable.