[Surgical Treatment for Pneumothorax with Granulomatosis with Polyangiitis].

A 42-year-old man had been treated for granulomatosis with polyangiitis for eight years. He was referred to our department with the diagnosis of right pneumothorax on chest radiograph. After chest drainage, the surgical treatment was performed because of continuing air leak from chest tube. Under thoracoscopic approach, the pleural adhesions were carefully dissected and the air leak site was sutured and enforced by a polyglycolic acid sheet with fibrin glue. No recurrence of pneumothorax was observed six months after surgery.

[Necrotizing Fasciitis and Frostbite of the Male External Genitalia : A Case Repor].

We report a case diagnosed as both necrotizing fasciitis and frostbite of the male external genetalia. The patient was a 58-year-old man with diabetes mellitus. He was referred to our hospital with swelling of his penis and icing there for 3 weeks. We diagnosed his condition as necrotizing fasciitis of the external genitalia and decided to perform an emergency operation. Although we consulted plastic surgeons about debridement of a large area, they considered that there was no necessity for it because the scrotum, perineum and abdominal wall were frostbitten. The final clinical diagnosis was necrotizing fasciitis of the penile region and frostbite around the perineal region. We performed partial penectomy without debridement. Through collaboration with another medical department it was thus possible to avoid unnecessary invasive treatment.

Efficacy of prolonged ingestion of Lactobacillus acidophilus L-92 in adult patients with atopic dermatitis.

To evaluate the safety and efficacy of prolonged ingestion of Lactobacillus acidophilus L-92 (L-92) on skin symptoms in adult atopic dermatitis (AD) patients, a placebo-controlled double-blinded parallel-group comparison study was performed. This included daily administration of heat-killed and dried L-92 or placebo for 24wk in 50 AD patients who were 16yr old or older. The severity of skin symptoms was evaluated at baseline and at 4, 8, 12, 16, 20, and 24wk during the intervention using the investigator global assessment, eczema area and severity index, and scoring atopic dermatitis. Serum cytokine and blood marker levels were also measured at baseline and at 4, 8, 16, and 24wk during the intervention. No adverse events were reported during the study period. Compared with the placebo group, the L-92 group showed significant decreases in investigator global assessment, eczema area and severity index, and scoring atopic dermatitis. Subjective symptoms in adult AD patients were reduced by intake of L-92. Furthermore, it was suggested that sustained ingestion of L-92 resulted in suppression of scratching behavior and maintenance of remission status of skin symptoms. Sixteen weeks after the study commenced, a significant decrease in lactate dehydrogenase and a significant increase in transforming growth factor-ß were observed in the L-92 group compared with the placebo group. In the L-92 group, a significant elevation of IL-12 (p70) level at the end of treatment period compared with before the treatment was observed. This study suggested that L-92 suppresses type-2-helper-T-cell-dominant inflammation by activating regulatory T cells and type 1 helper T cells.

Acrylated Hyaluronic-Acid Based Hydrogel for the Treatment of Craniofacial Volumetric Muscle Loss.

Current treatment options for craniofacial volumetric muscle loss (VML) have disadvantages and cannot fully restore normal function. Bio-inspired semisynthetic acrylated hyaluronic acid (AcHyA) hydrogel, which fills irregularly shaped defects, resembles an extracellular matrix, and induces a minimal inflammatory response, has shown promise in experimental studies of extremity VML. We therefore sought to study AcHyA hydrogel in the treatment of craniofacial VML. For this, we used a novel model of masseter VML in the rat. Following the creation of a 5 mm × 5 mm injury to the superficial masseter and administration of AcHyA to the wound, masseters were explanted between 2 and 16 weeks postoperatively and were analyzed for evidence of muscle regeneration including fibrosis, defect size, and fiber cross-sectional area (FCSA). At 8 and 16 weeks, masseters treated with AcHyA showed significantly less fibrosis than nonrepaired controls and a smaller decrease in defect size. The mean FCSA among fibers near the defect was significantly greater among hydrogel-repaired than control masseters at 8 weeks, 12 weeks, and 16 weeks. These results show that the hydrogel mitigates the fibrotic healing response and wound contracture. Our findings also suggest that hydrogel-based treatments have potential use as a treatment for the regeneration of craniofacial VML and demonstrate a system for evaluating subsequent iterations of materials in VML injuries.

Anti-influenza virus effects of both live and non-live Lactobacillus acidophilus L-92 accompanied by the activation of innate immunity.

The antiviral effects of both a live and non-live Lactobacillus acidophilus strain L-92 (L-92) were investigated by oral administration (10 mg/mouse per d) daily for 21 d in a mouse model infected intranasally with influenza virus (H1N1). Virus titres in the lung of mice administered either live or non-live L-92 cells daily for 15 d were repressed 6 d after virus infection compared with the control group. Natural killer (NK) activity in the orally administered non-live L-92 group was higher compared with that of the control group before virus infection and on day 6. In contrast, NK activity in the live L-92 group compared with the control group was not significantly changed on both days, but was significantly higher on day 1. In contrast, live L-92 showed a greater repression of virus proliferation compared with non-live L-92, 6 d after the infection. Live L-92 decreased the number of neutrophils in the lung and suppressed lung weight, leading to the consequent deterioration of consolidation scores of the lung. These results indicated that pretreatment of live or non-live L-92 cells had protective effects against influenza virus infection. Among the measured cytokines and chemokines, eotaxin, macrophage colony-stimulating factor, IL-1b, RANTES (regulated on activation, normal T cell expressed and secreted) and interferon-a were significantly increased in the lung: IL-17 was significantly increased in Peyer's patch of the live L-92 group compared with the control group. A mechanistic study suggested that the enhancement of NK activity in the lung caused by stimulating various antiviral cytokines and chemokines after the oral administration of L-92 cells might be important in protecting against virus infection.

Surgical Management of Breast Cancer Developing Along the Pathway of a Ventriculoperitoneal Shunt: A Case Report.

BACKGROUND Ventriculoperitoneal shunts are commonly used in neurosurgery for the treatment of hydrocephalus. This case report details an unusual instance where breast cancer developed along the pathway of an existing ventriculoperitoneal shunt. CASE REPORT An 86-year-old woman, who previously underwent ventriculoperitoneal shunt placement for normal-pressure hydrocephalus, visited our hospital upon detecting a mass in her left breast. The physical examination discovered an irregular mass located at the 9 o'clock position of the left breast. Subsequent breast ultrasonography identified a 36 mm mass with indistinct borders, rough margins, and signs of skin infiltration. Invasive ductal carcinoma of a triple-negative subtype was diagnosed through a core-needle biopsy. Contrast-enhanced computed tomography indicated the ventriculoperitoneal shunt's pathway, running from the left ventricle, passing through the center of the breast mass, and leading into the abdominal cavity. Fears of shunt occlusion and potential infection due to the untreated breast cancer prompted surgical intervention after consultation with the neurosurgeon. The surgery involved rerouting the ventriculoperitoneal shunt from the left thoracoabdomen to the right, performing a left mastectomy, and removing the fistula in the abdominal wall to minimize the risk of cancer recurrence along the shunt pathway. Postoperative histopathological examination confirmed the initial diagnosis of invasive ductal carcinoma of a triple-negative subtype, with no malignancy detected in the removed abdominal wall fistula. CONCLUSIONS Taking into account prior cases of cancer metastasizing distantly due to ventriculoperitoneal shunts, our case emphasizes the necessity to consider additional preventative measures against cancer seeding. This approach is particularly significant when treating breast cancer that arises along the pathway of a ventriculoperitoneal shunt, apart from performing conventional breast cancer surgery.

[Prediction of serum albumin levels by non-invasive factors among elderly female patients].

AIM: To apply nutrition care management to elderly female patients, we predicted serum albumin (s-Alb) levels by non-invasive factors. METHODS: After excluding patients with lesions/diseases which were directly related to s-Alb levels, we investigated 147 elderly women aged 75-years or over who were taking meals orally and were hospitalized from April 2008 to April 2009 at a hospital in Toyota. The patients were classified into 2 groups, one of patients with s-Alb levels of 3.5 g/dl or below (n=80), and the other of those with s-Alb levels of over 3.5 g/dl (n=67). Between the 2 groups, we examined differences in age, body mass index (BMI), living arrangements, necessary nursing care level (NNCL), bed confinement level (BCL), OH scale level (OHSL), and dietary intake either by the Student t-test, Mann-Whitney U test or chi-square test. Pearson correlation coefficients were calculated among s-Alb levels and selected variables. Taking into account the correlation coefficients, we conducted multiple regression analysis adopting the s-Alb level as a dependent variable and non-invasive factors as independent variables. For all the performed tests and analyses, a p value of less than 0.05 (on two-tailed analysis) was assumed to represent a statistically significant difference. RESULTS: S-Alb level was significantly associated with variables, including age, BMI, NNCL, BCL, OHSL, and percentage of protein intake (PPI). Multiple regression analysis revealed 4 significant variables: age, BCL, OHSL, and PPI. The multiple regression equation was y=4.977-(0.098×OHSL)-(0.080×BCL)-(0.016×age)+(0.003×PPI), and the multiple correlation coefficient R(2) was 0.398 (p <0.001). CONCLUSIONS: S-Alb levels among elderly female patients may be predicted by 4 non-invasive variables: age, BCL, OHSL, and PPI.

New Concept for Severely Burned Ear Reconstruction: Grafting of the Frame Using Banked Ear Cartilage Combined with Costal Cartilage.

Damage and loss of ear cartilage can easily occur in a burned ear accompanied by severe extensive burns due to exposure of the cartilage or chondritis. Deformity can also occur due to later development of scar contracture despite minimal damage to the ear cartilage when the injury occurred. We have developed a new technique for treatment of a deep burn in the whole ear. METHODS: In the new technique, the ear is amputated, and soft tissues are denuded. The cartilage is banked in a subcutaneous pocket in the chest and later regrafted in combination with costal cartilage. RESULTS: Although techniques for regrafting banked ear cartilage have been reported, the reconstructed ear cannot acquire a good shape because of absorption of the cartilage or lack of intensity to sustain the outline of the ear. Meanwhile, when we tried to reconstruct an ear by only using costal cartilage, we found it difficult to fabricate a frame because most patients are adults in whom the costal cartilage is too rigid and fragile to be shaved or combined. In our technique, the frame has both the advantages of sufficient intensity in costal cartilage and a smooth curved surface together with elasticity in the ear cartilage. Ears reconstructed by our technique have a natural appearance. CONCLUSION: Our technique can be used for cases in which treatment for another large area of the body surface needs to be performed first to save the patient's life.

Vacuum membrane distillation by microchip with temperature gradient.

A multilayered microchip (25 x 95 mm) used for vacuum distillation is designed, fabricated and tested by rectification of a water-methanol mixture. The polymer chip employs a cooling channel to generate a temperature gradient along a distillation channel below, which is separated into a channel (72 microm deep) for liquid phase and a channel (72 microm deep) for vapor phase by an incorporated microporous poly(tetrafluoroethylene) (PTFE) membrane. The temperature gradient is controlled by adjusting hotplate temperature and flow rate of cooling water to make the temperatures in the stripping section higher than the increasing boiling points of the water-enriched liquids and the temperatures in the rectifying section lower than the decreasing dew points of the methanol-enriched vapors. The effects of temperature gradient, feed composition, feed flow rate and membrane pore size on the micro distillation are also investigated. A theoretical plate number up to 1.8 is achieved at the optimum conditions.

Palladium nanoparticles captured in microporous polymers: a tailor-made catalyst for heterogeneous carbon cross-coupling reactions.

A new strategy based on polymerization-induced phase separation (PIPS) techniques was proposed for fabricating palladium nanoparticles (PdNPs) captured in a microporous network polymer. Pd(OAc)(2) was premixed with a monomer having a poly(amidoamine)-based dendrimer ligand, and subsequently this was thermally polymerized with an excess amount of ethylene glycol dimethacrylate under PIPS conditions. In this system, the formation of PdNPs occurred concurrently with the polymer synthesis in a one-pot process, even with no additional reducing reagent. The resultant microporous polymer was found to have a mesoporosity; the nitrogen sorption analysis gave a specific-surface area of 511 m(2) g(-1), an average pore diameter of 9.9 nm, and a total pore volume of 1.01 mL g(-1). The TEM images of the polymer revealed that the created PdNPs were very small with a diameter of mainly ca. 2.0 nm; the high-resolution images were lattice-resolvable, showing the crystalline nature of the PdNPs (Pd(111) facets). Catalytic performances of the PdNP-containing microporous polymers were investigated for a heterogeneous Suzuki-Miyaura reaction of 4'-bromoacetophenone and phenylboronic acid in water. In the presence of 10(-2) molar equiv of the polymer, the reaction efficiently proceeded at 80 degrees C and gave the desired product, 4-acetylbiphenyl, in >90% yield after 2 h. On the basis of the ICP-AES analysis, the Pd content released into the solution phase was estimated to be only 0.27% of the initial charge. Thereby, this polymer was successfully recovered by simple filtration and reused with only a minimal loss of activity (yield >90% even at the eighth run). When the catalytic reaction was examined with a low amount of the polymer catalyst, the turnover number (TON) reached 8.5 x 10(4) while maintaining a good yield. Finally, the dendrimer template effect of the polymer catalyst was discussed by referring to the catalytic performances of a control polymer prepared with nonintegrated ligand monomers.

The Laterally Extended Paramedian Forehead Flap for Nasal Reconstruction: The Delay Technique Revisited.

BACKGROUND: Problems with poor circulation often occur when a large defect or a distant region, such as the apex of the nose, is covered with a paramedian forehead flap. Delay technique increases the safety of reconstruction procedures, but it has been used less frequently because a 2-stage surgery is necessary, and various other flaps and techniques have been developed. METHOD: We performed the delay technique of paramedian forehead flap at the same time as tumor resection. For the flap, a narrow pedicle of about 1-cm was prepared on the supratrochlear artery and vein, and the incision was extended toward the lateral side conforming to the defect morphology, and a paramedian forehead flap with a design consistent with the esthetic unit containing the defect was prepared. The region below the flap was dissected to create the flap bipedicle, and surgery was completed. RESULT: This procedure was used in 4 patients with malignant tumor of the external nose, and the flap survived perfectly in all patients. The postoperative esthetic outcome was also found to be good. CONCLUSIONS: This procedure does not increase the frequency of surgery, circulation in the flap is maintained, the flap pedicle on the supratrochlear artery can be made narrow, and flap thinning can be performed from the beginning. Coverage of an extensive defect is possible because a large flap can be excised, and satisfactory esthetic appearance can be obtained by matching with the esthetic unit. The delay technique for various flaps (not limited to forehead flap alone) should be considered an effective technique for the current treatment of malignant tumors.

Vacuum membrane distillation on a microfluidic chip.

A multilayered microfluidic chip used for vacuum membrane distillation is designed, fabricated and tested by rectification of a water-methanol mixture.

Dementia Cafés as a Community Resource for Persons With Early-Stage Cognitive Disorders: A Nationwide Survey in Japan.

OBJECTIVES: Dementia cafés are expected to serve as a new community resource based on the national dementia strategy in Japan. The objective of the present study was to examine effective ways to manage dementia cafés through an overview of dementia cafés in Japan and an analysis of the factors related to their effectiveness on attendees. DESIGN: Cross-sectional analysis. SETTING AND PARTICIPANTS: Representatives of 1477 dementia cafés in Japan. MEASURES: Questionnaires regarding the dementia cafés' characteristics, management members, staff, their guests and effectiveness on people with dementia, their families, and community members were sent to the cafés, with instructions to have them completed by the café representatives. Logistic regression analysis was performed with the effect on each guest attribute as a dependent variable, and factors related to the effectiveness of dementia cafés were analyzed. RESULTS: Questionnaires were sent to a total of 2728 dementia cafés in Japan, and responses were received from 1477 (54.1%). The most common meeting frequency and meeting hours were once a month (64.8%) and 2 hours (53.8%), respectively. Analysis of the effectiveness of dementia cafés on 3 groups of guests indicated the following preferences for café program: people with dementia preferred frequent meetings and activities, families of people with dementia preferred having a place for private consultation and advice and peer meetings, and community members preferred frequent meetings and both mini-lectures and private consultation and advice. Logistic regression analysis further revealed that all types of guests preferred the presence of the same types of guests. CONCLUSIONS/IMPLICATIONS: This study revealed the multicomponent nature of dementia cafés. The results suggest that a good balance of programs and guests would enhance the cafés' effectiveness among the multiple stakeholders in regard to dementia, especially in the early stage of the disease.

The membrane phospholipid cardiolipin plays a pivotal role in bile acid adaptation by Lactobacillus gasseri JCM1131T.

Bile acids exhibit strong antimicrobial activity as natural detergents, and are involved in lipid digestion and absorption. We investigated the mechanism of bile acid adaptation in Lactobacillus gasseri JCM1131T. Exposure to sublethal concentrations of cholic acid (CA), a major bile acid in humans, resulted in development of resistance to otherwise-lethal concentrations of CA by this intestinal lactic acid bacterium. As this adaptation was accompanied by decreased cell-membrane damage, we analyzed the membrane lipid composition of L. gasseri. Although there was no difference in the proportions of glycolipids (~70%) and phospholipids (~20%), adaptation resulted in an increased abundance of long-sugar-chain glycolipids and a 100% increase in cardiolipin (CL) content (to ~50% of phospholipids) at the expense of phosphatidylglycerol (PG). In model vesicles, the resistance of PG vesicles to solubilization by CA increased with increasing CL/PG ratio. Deletion of the two putative CL synthase genes, the products of which are responsible for CL synthesis from PG, decreased the CL content of the mutants, but did not affect their ability to adapt to CA. Exposure to CA restored the CL content of the two single-deletion mutants, likely due to the activities of the remaining CL synthase. In contrast, the CL content of the double-deletion mutant was not restored, and the lipid composition was modified such that PG predominated (~45% of total lipids) at the expense of glycolipids. Therefore, CL plays important roles in bile acid resistance and maintenance of the membrane lipid composition in L. gasseri.

Establishment of a Standardized Technique for Concha-type Microtia-How to Incorporate the Cartilage Frame into the Remnant Ear.

We have already reported surgical procedures for lobule-type microtia that provide an excellent contour and shape of the ear with minimum sacrifice of the donor. We have succeeded in establishing a standard surgical technique for almost all types of concha-type microtia that effectively uses the remnant ear and can use a unified costal cartilage frame. METHODS AND RESULTS: The concept of our technique is that remnant cartilage should be used maximally but that the deformed area should be completely replaced by the costal cartilage frame. The differences between the cartilage frame for lobule-type microtia and that for concha-type microtia are that the lower half beneath the antihelical area and the concha cymba in the base frame are omitted in concha-type microtia. The area from the tragus to the incisura of the tragus in the antihelical-tragal frame is also omitted. The area of the helical crus in the helical frame and the lower half in the antihelix are not immobilized in the base frame and are free edges. On the other hand, the remnant cartilage outside the concha is removed, but the antitragus is preserved. When the cartilage frame and the remnant are incorporated, all of the components of the ear can be provided. CONCLUSION: The ears created by our technique have a natural appearance and clear contour.

A novel CD4-conjugated ultraviolet light-activated photocatalyst inactivates HIV-1 and SIV efficiently.

In this study, we found that the electric potential derived from the redox reaction of ultraviolet (UV)-illuminated CD4-conjugated titanium dioxide (TiO2) inactivated a wide range of high-titered primary HIV-1 isolates, regardless of virus co-receptor usage or genetic clade. In vitro incubation of HIV-1 isolates with CD4-conjugated TiO2 (CD4-TiO2) followed by UV illumination led to inhibition of viral infectivity in both H9 cells and peripheral blood mononuclear cells as well as to the complete inactivation of plasma virions from HIV-1-infected individuals. Treatment with a newly established extra-corporeal circulation system with the photocatalyst in rhesus macaques completely inactivated plasma virus in the system and effectively reduced the infectious plasma viral load. Furthermore, plasma viremia and infectious viral loads were controlled following a second therapeutic photocatalyst treatment during primary SIV(mac239) infection of macaques. Our findings suggest that this therapeutic immunophysical strategy may help control human immunodeficiency viral infection in vivo.

New technical method to correct secondary vermilion deformities with cleft lip.

Various techniques for correcting whistling deformities that occurred after primary surgery for cleft lip have been reported. These techniques are mainly intended to correct the lack of volume of the red lip. However, irregularity of the dry-wet lip junction (mucocutaneous junction) in the red lip has rarely been mentioned. If the wet lip is located in an exposed area, not only is the aesthetic appearance poor but also uncomfortable complications such as a crusted or bleeding lip repeatedly occur under a dry condition. A new technique for correcting the irregular line of the dry-wet lip junction is described in this report. The technique is simple. After removal of the exposed wet lip, flaps are designed on both dry lip sides of the defect as M-W-M plasty and are transposed toward the defect. The dog-ears are small; the scar is inconspicuous because it is incorporated with the wrinkle line, and scar contracture is prevented. In addition, more soft tissues may be included to correct a mild whistling deformity.

Subcutaneous pedicle V-Y flap for release of incomplete congenital syndactyly.

BACKGROUND: Various procedures for correction of congenital syndactyly of hand or foot have been described. For incomplete syndactyly, some of the reported techniques use only local flaps from surrounding tissues. A novel technique for the correction of incomplete syndactyly, using a dorsal triangular flap and two palmar small flaps, is described in this article. METHODS: A triangular flap is first marked on the affected web space. The size of the flap should be the same as the unaffected side or other web space. Then a straight line is marked from the proximal apex of the triangle to the level of the metacarpophalangeal (MP) joint. After full skin incision, minimal peripheral undermining is done, and the triangular flap is transposed proximally, as in the Y-V advancement procedure, and sutured. Then two incisions are made from the distal part of the flap, transposing small flaps as in the five-flap method, and closed primarily. RESULTS: We treated ten cases of congenital syndactyly of the hand or foot. We were able to correct a good web space without skin grafting in all cases. CONCLUSION: The design for our technique is simple, and the technique can be performed easily. The operation can be performed in a short time, the blood supply of the flap is preserved, the flap has a wide range of motion, and a deep and smooth dorsal slope is produced. This technique is an attractive alternative to previously reported methods for syndactyly correction.

Lactobacillus acidophilus CP23 with weak immunomodulatory activity lacks anchoring structure for surface layer protein.

To determine the reason for the low levels of Surface layer protein A (SlpA) on CP23 cells, which might play a crucial role in the immunomodulatory effect of Lactobacillus acidophilus, the DNA sequence of the slpA gene of CP23 and L-92 strains, including the upstream region, were analyzed. Unexpectedly, there was no significant difference in the predicted amino acid sequence of the C-terminus needed for cell anchoring, and only an additional Ala-Val-Ala sequence inserted in the N-terminal region of the mature CP23 protein. Therefore, anchoring of SlpA on the cell wall of CP23 and L-92 was evaluated by a reconstitution assay, which showed that SlpA released by LiCl treatment from both CP23 and L-92 was successfully anchored on LiCl-treated L-92 cells, but not on LiCl-treated CP23 cells. Moreover, quantitative analysis of SlpA protein in the culture medium of CP23 and L-92 by ELISA revealed higher levels of SlpA secretion in CP23 cells than in L-92 cells. Collectively, these results suggest that the lower levels of SlpA on the surface of CP23 cells might be caused by less cell wall capacity for SlpA anchoring, leading to an accumulation of SlpA in the culture medium of CP23 cells. The present study supports the importance of cell surface structure of L. acidophilus L-92 for SlpA anchoring on the cell surface needed for immunomodulatory effect.

A pulmonary metastatic model of human non-small cell lung carcinoma cells that produce a neutrophil elastase-like molecule in severe combined immunodeficiency mice.

STUDY OBJECTIVES: To establish a clinically relevant animal model of pulmonary metastases of human non-small cell lung carcinoma (NSCLC) cells in severe combined immunodeficiency (SCID) mice, which can be used for repetitive investigations, so as to improve our understanding and management of the cellular and molecular mechanisms of human lung cancer metastases. METHODS AND RESULTS: SCID mice subcutaneously injected in the flank with 1 x 10(6) EBC-1 cells derived from human lung squamous cell carcinoma were killed weekly for examination until 12 weeks after tumor inoculation. The biological characteristics of implanted tumors and their metastatic foci were investigated by hematoxylin-eosin staining and immunostaining for neutrophil elastase (NE). Three weeks after ectopic implantation, EBC-1 cell lines formed a tumor at the inoculation site and grew steadily to show a plateau at 10 weeks. EBC-1 cells formed multiple metastases in the lung 7 weeks after tumor inoculation; their numbers increased steadily until 12 weeks in all mice. Immunoreactivity for NE was intense in the metastatic tumor cells. Then, to establish the primary tumor amputation/pulmonary metastasis model and to evaluate how primary tumor amputation influences the development of pulmonary metastases at the cellular and molecular level, excision was performed before (3 weeks and 5 weeks after inoculation) and after (7 weeks and 9 weeks after inoculation) formation of lung metastases. When the primary tumor was excised 3 weeks after tumor inoculation, all mice had pulmonary metastasis at 12 weeks after inoculation. Blood samples obtained at 3 weeks after tumor inoculation contained human beta-actin messenger RNA, which represents circulating tumor cells. CONCLUSION: Our NSCLC EBC-1 pulmonary metastasis model is reliable, technically simple, and predictably results in pulmonary metastasis from early hematogenous spread. This model may be useful for elucidating the mechanism of pulmonary metastasis in human lung cancer, and testing anti-metastatic efficacy of therapeutic agents in vivo.