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In the present study, we studied the feasibility of deleting essential genes in insect cells by using bacmid and purifying recombinant bacmid in Escherichia coli DH10B cells. To disrupt the orf4 (open reading frame 4) gene of BmNPV [Bm (Bombyx mori) nuclear polyhedrosis virus], a transfer vector was constructed and co-transfected with BmNPV bacmid into Bm cells. Three passages of viruses were carried out in Bm cells, followed by one round of purification. Subsequently, bacmid DNA was extracted and transformed into competent DH10B cells. A colony harbouring only orf4-disrupted bacmid DNA was identified by PCR. A mixture of recombinant (white colonies) and non-recombinant (blue colonies) bacmids were also transformed into DH10B cells. PCR with M13 primers showed that the recombinant and non-recombinant bacmids were separated after transformation. The result confirmed that purification of recombinant viruses could be carried out simply by transformation and indicated that this method could be used to delete essential genes. Orf4-disrupted bacmid DNA was extracted and transfected into Bm cells. Viable viruses were produced, showing that orf4 was not an essential gene.

A leucine aminopeptidase was purified for the first time from marine fish red sea bream ( Pagrus major) skeletal muscle to homogeneity with 4850-fold and a yield of 7.4%. The purification procedure consisted of ammonium sulfate fractionation and chromatographies including DEAE-Sephacel, Sephacryl S-200, hydroxyapatite, and phenyl-Sepharose. The enzyme was approximately 96 kDa as estimated by SDS-PAGE and gel filtration and preferentially hydrolyzed substrate Leu-MCA. The enzymatic activity was optimal at 45 degrees C and pH 7.5. The K m and k cat values of the enzyme for Leu-MCA were 1.55 microM and 26.4 S (-1) at 37 degrees C, respectively. Activation energy ( E a) of the enzyme was 59.6 kJ M (-1). The enzyme was specifically inhibited by metal-chelating agents, and Zn (2+) and (or) Mn (2+) seemed to be its metal cofactor(s). In addition, bestatin strongly inhibited its activity, and K i was 1.44 microM. Using a highly specific polyclonal antibody, the location of enzyme was demonstrated intracellularly and distributed in different tissues.

Two trypsins of anionic form (trypsin A) and cationic form (trypsin B) from the pyloric caeca of mandarin fish (Siniperca chuatsi) were highly purified by a series of chromatographies, including DEAE-Sephacel, Sephacryl S-200 HR, Q-Sepharose or SP-Sepharose. Purified trypsins revealed a single band on native-PAGE. The molecular weights of trypsin A and B were 21kDa and 21.5kDa, respectively, as estimated by SDS-PAGE, both under reducing and non-reducing conditions. Zymography analysis showed that both trypsins were active in degrading casein. Trypsin A and B exhibited maximal activity at 35°C and 40°C, respectively, and shared the same optimal pH of 8.5, using Boc-Phe-Ser-Arg-MCA as substrate. The two trypsins were stable up to 45°C and in the pH range from 4.5 to 11.0. Trypsin inhibitors are effective on these two enzymes and their susceptibilities were similar. Both trypsins were activated by metal ions such as Ca(2+) and Mg(2+) and inactivated by Fe(2+), Zn(2+), Mn(2+), Cu(2+), Al(3+), Ba(2+) and Co(2+) to different degrees. Apparent Km values of trypsin A and B were 2.18μM and 1.88μM, and Kcat values were 81.6S(-1) and 111.3S(-1) for Boc-Phe-Ser-Arg-MCA, respectively. Immunoblotting analysis using anti-common carp trypsin A positively cross-reacted with the two enzymes, suggesting their similarity. The N-terminal amino acid sequence of trypsin B was determined as IVGGYECEAH, which is highly homologous with trypsins from other species of fish., (Copyright © 2008 Elsevier Ltd. All rights reserved.)

The impact of W-Beijing genotype Mycobacterium tuberculosis on treatment outcome was evaluated in 249 newly diagnosed tuberculosis patients. No significant difference in the treatment outcome was found between the W-Beijing and non-W-Beijing groups. However, a poor outcome was more common in the elderly patients (>or=65 years) infected with the W-Beijing strain.

Four pepsinogens (PG-I, PG-II, PG-III(a), and PG-III(b)) were highly purified from the stomach of the freshwater fish mandarin fish (Siniperca chuatsi) by ammonium sulfate fractionation, anion exchange, and gel filtration. The molecular masses of the four purified PGs were 36, 35, 38, and 35 kDa, respectively. All the pepsinogens converted into their active form pepsins within a few minutes under pH 2.0. The optimum pH and temperature of the four enzymes were 3.0-3.5 and 45-50 degrees C, using hemoglobin as a substrate. The N-terminal amino acid sequences of PG-I and PG-II were determined to the 12th and 17th amino acid residues, respectively. Western blot analysis using antisea bream polyclonal antibodies cross reacted with PG-I, PG-II, and PG-III(b) while no cross reaction with PG-III(a) was detected, suggesting the diversity of pepsinogens in fish.

Objective: To assess the effects of the alteration of humidity and (or) temperature on weight of filters without and with ambient particulate matter in a balance room., Methods: The mass of blank dust sampling filters were weighed under (18 +/- 1) degrees C and (28 +/- 1) degrees C respectively, with the humidity varying from 35% relative humidity (RH) to 100% RH in a balance room. Then the blank filters were divided into two groups and were used to sample total dust and respirable dust. After sampling, the loaded filters were re-weighed under above conditions and the mass difference before and after the sampling were compared and analyzed., Results: The vibration of the average mass of filters varied from 0.10 to 0.13 mg and from 0.06 to 0.09 mg under the temperatures of (18 +/- 1) degrees C and (28 +/- 1) degrees C respectively; When both the temperature and humidity changed, it varied from 0.12 to 0.16 mg. The deviation of average mass difference ranged from 0.07 to 0.10 mg and from 0.04 to 0.08 mg under the two temperatures mentioned above; When both the temperature and humidity changed, it varied from 0.09 to 0.14 mg. The average mass of blank filters and loaded filters were all positively correlated with the change of humidity (P < 0.01). No effects of humidity on the average mass difference of the loaded filters were observed. The average mass differences of loaded filters and blank filters under (18 +/- 1) degrees C were significantly higher than that under (28 +/- 1) degrees C (P < 0.01) when humidity was not changed., Conclusion: The alteration of humidity and (or) temperature in a balance room attributes to the deviation of the measurement of the mass of filters and thus affects the gravimetric measurements of ambient particulate matter.

Aim: To explore the efficacy of hospitals using case management with Directly Observed Treatment - Short course (DOTS) to monitor the adherence of patients with pulmonary tuberculosis in Taiwan., Background: Non-adherence to anti-tuberculosis chemotherapy is the major problem in treating patients with tuberculosis. Community-based case management coupled with DOTS has been applied to patients with tuberculosis and has resulted in good results in some countries. Taiwan has a high incidence of tuberculosis, and although it has implemented DOTS, the expected increased efficacy has not yet been realized., Design and Methods: The study used a quasi-experimental design. Using age and gender as matching factors, 96 subjects were randomly assigned to one of three groups in 2002-2003. Experimental group I was to receive DOTS case management comprising in-hospital education, direct daily observation in the first two months and one home visit per week. Experimental group II received traditional case management comprising in-hospital education and one home visit per month. The control group did not receive any intervention., Results: The adherence, the rate of completion, the treatment success, sputum conversion and chest X-ray improvement of experimental group I were significantly improved compared with experimental group II and the control group. The completion rate in experimental group I was higher than the general rate for Taiwan during the past six years and the treatment success rate met the standards of the World Health Organization., Conclusion: Hospitals using case management with DOTS can improve the adherence of tuberculosis patients and the control of tuberculosis-epidemic situations. Relevance to clinical practice. In a rapidly changing healthcare environment, clinical nurses can make a significant contribution to healthcare delivery for tuberculosis patients. This study has provided further insight into the implementation of hospital-to-community level case management using DOTS by nurses.

Gelatinolytic proteinases from common carp dark muscle were purified by 30-60% ammonium sulfate fractionation and a combination of chromatographic steps including ion exchange on DEAE-Sephacel, gel filtration on Sephacryl S-200, ion exchange on High-Q, and affinity on gelatin-Sepharose. The molecular masses of these proteinases as estimated by SDS-PAGE were 75, 67, and 64 kDa under nonreducing conditions. The enzymes revealed high activity at a slightly alkaline pH range, and their activities were investigated using gelatin as substrate. Metalloproteinase inhibitors, EDTA, EGTA, and 1,10-phenanthroline, almost completely suppressed the gelatinolytic activity, whereas other proteinase inhibitors did not show any inhibitory effect. Divalent metal ion Ca (2+) is essential for the gelatinolytic activity. Furthermore, these gelatinolytic proteinases hydrolyze native type I collagen effectively even at 4 degrees C, strongly suggesting their involvement in the texture softening of fish muscle during the post-mortem stage.

Background/purpose: This retrospective study was conducted to evaluate the drug resistance patterns of Mycobacterium tuberculosis in a medical center in northern Taiwan between 2003 and 2004 in comparison to those reported in 1990-1992., Methods: A total of 611 non-duplicate M. tuberculosis isolates from culture-proven tuberculosis (TB) cases were tested for drug susceptibility against five first-line anti-TB drugs in a clinical mycobacterial laboratory using the agar proportional method for isoniazid (INH), rifampicin (RIF), ethambutol (EMB), and streptomycin (SM). The Wayne assay, which measures the activity of pyrazinamide (PZA), was used for PZA susceptibility testing., Results: Of 611 patients, including 510 males and 101 females, 70.2% of patients were older than 65 years. A total of 339 isolates (55.5%) were resistant to one or more drugs. Isolates from patients aged <25 years showed a significantly higher drug resistance rate (79.2%) compared with other age groups (p=0.0312). Single-drug resistance was observed in 97 (15.9%) of all isolates. Monoresistance to PZA (8.0%) was most frequent, followed by INH (5.1%), RIF (0.5%), EMB (1.6%), and SM (0.7%). Among the polydrug resistant isolates (PDR-TB), resistance rates were 35.5% for INH and 27.0% for RIF. One hundred and fifty-nine isolates (26.0%) were resistant to both INH and RIF (multidrug-resistant [MDR] TB); 94.6% of RIF-resistant isolates were also resistant to INH. The overall drug resistance rates and percentages of PDR-TB and MDR-TB increased over the 12-year study period (p<0.001). Based on medical records, primary cases were identified in 486 (84.7%) out of 574 patients, and resistance to any drug was identified in 268 (55.1%) patients, of which 130 (26.7%) were MDR-TB. Among the 88 with recurrent TB, 54 (61.4%) were resistant to at least one drug, and MDR-TB was identified in 29 (33.0%) patients. A history of previous anti-TB therapy was a significant factor for overall drug resistance, PZA monoresistance, PDR-TB, and MDR-TB (p<0.001)., Conclusion: The emergence of M. tuberculosis isolates resistant to anti-TB agents in this hospital, and in particular among young patients, is alarming. Strict measures to control and prevent drug-resistant TB are urgently needed.

Objectives: Rhabdomyoma, the most common primary fetal cardiac tumor, is often associated with tuberous sclerosis (TS). We aimed to evaluate outcome in cases diagnosed with fetal cardiac rhabdomyoma., Methods: This study presents 11 cases with fetal cardiac rhabdomyoma. In addition, all relevant published cases of antenatally diagnosed cardiac rhabdomyoma since 1982 were identified from MEDLINE. We evaluated the following risk factors associated with clinical impact and perinatal outcome: family history of TS, gestational age at diagnosis, tumor size, site and number of tumors, tumor progression, and associated intracardiac and extracardiac anomalies., Results: In this meta-analysis, 138 cases, including nine newly added by us, were categorized into Group A (107 live babies) and Group B (16 neonatal deaths and 15 intrauterine fetal deaths). Univariate analysis showed that large cardiac tumors (P < 0.0001), fetal dysrhythmia (P < 0.0001) and hydrops (P < 0.0001) were strong predictors of neonatal outcome. Tumor size >or= 20 mm (relative risk (RR), 20.6; 95% CI, 2.2-195.9; P = 0.009) and fetal dysrhythmia (RR, 13.6; 95% CI, 2.9-62.3; P = 0.001) were significantly associated with neonatal morbidity. TS, present in 85/133 (63.9%) cases, was significantly associated with multiple cardiac tumors (P < 0.0001) and family history of TS (P = 0.02)., Conclusions: Large tumor size and hydrops are significantly associated with poor neonatal outcome, whereas family history of TS and multiple fetal cardiac tumors are associated with TS. Any sonographic detection of a fetal cardiac tumor should warrant further investigation for the possible presence of associated disorders., ((c) 2008 ISUOG. Published by John Wiley & Sons, Ltd.)

Objective: To study pharmacokinetics of honokiol in Cortex of Magnolia officinalis and its compound prescription Houpu Sanwu Decoction (HPSWD) in rat, and discuss the change on pharmacokinetic process affected by other components., Methods: The rats were divided into two groups, one was supplied with HPSWD and the other was administrated with Cortex of Magnolia officinalis. Concentrations of honokiol in rat plasma were then determined using HPLC method and main pharmacokinetic parameters were estimated., Results: The plasma concentrations of honokiol of both groups were conformed to the two-compartment model with first order absorption. There existed significant differences in AUC, C(max), T(max), CL/F among Cortex of Magnolia officinalis and HPSWD., Conclusion: The results indicate that Rhubarb and Immature Orange Fruit in HPSWD can influence the asorption, distribution and elimination of honokiol.

Open reading frame 76 of Bombyx mori nucleopolyhedrovirus (BmNPV), designated as Bm76, is a gene whose function is completely unknown. With EGFP fused to the 3' terminal of Bm76 as the reporter gene and BmNPV bacmid as the expression vector, a recombinant bacmid was successfully constructed expressing Bm76-EGFP fusion protein under the control of polyhedrin promoter in Bombyx mori cells (Bm cells), BmNPV's permissive cell line, laying the foundation for rescue experiment of Bm76 deletion mutant. Moreover, the supernatant from Bm cells transfected with the recombinant bacmid was used to infect Trichoplusia Ni cells (Tn cells), BmNPV's non-permissive cell line. Unexpectedly, the expression of Bm76-EGFP fusion protein in some Tn cells was detected, implying that viral DNA was replicated in these cells. The causes are being studied for the inability of BmNPV to produce enough viable budded viruses in Tn cells despite of viral DNA replication.

Truncus arteriosus, a double aortic arch, and a cervical aortic arch are all rare cardiovascular anomalies. We experienced a unique female newborn with the rare combination of truncus arteriosus with a cervical double aortic arch, which probably resulted from abnormal persistence of the bilateral 2nd or 3rd rather than the 4th embryonic aortic arches and failure of regression of the right 8th somitic segment of the right dorsal aorta. She presented with respiratory distress soon after birth, which was initially attributed to the vascular ring and hypertensive pulmonary arteries. Our inability to relieve her respiratory compromise by surgical division of the vascular ring and main pulmonary artery banding prompted the diagnosis of left main bronchial compression caused by a posteriorly displaced dilated ascending aorta that compressed the right pulmonary artery and left main bronchus against the descending aorta. The patient then underwent successful left main bronchus stent implantation. We speculate the cervical double aortic arch is redundant in nature and is a loose ring that may not cause tracheal compression. Nevertheless, a posteriorly displaced dilated ascending aorta in patients with truncus arteriosus may compress the right pulmonary artery and the main bronchus on the side of the aortic arch against the descending aorta., ((c) 2008 S. Karger AG, Basel.)

Purpose: To evaluate and compare the health-promoting behavior of adolescents with congenital heart disease (CHD) to that of adolescents without CHD., Methods: Cross-sectional data were collected from pediatric cardiology outpatient departments at two medical centers in Taiwan. A total of 1209 adolescents, including 316 with various forms of CHD and 893 without CHD, completed the Adolescent Health Promotion (AHP) scale. Of those with CHD, 162 were female, and 12-18 years old. The scores of adolescents with CHD were compared with published normative adolescent data., Results: No significant differences were found between those adolescents with CHD and those without in terms of dimensions of the AHP, which consisted of nutrition, social support, health responsibility, life appreciation, exercise, stress management, and overall health-promoting behavior. The three highest and lowest mean scores of scale items between these two groups were identified. Factors among adolescents with CHD, such as age, gender, parental educational level, and cardiac function were significantly associated with at least one dimension of the AHP. Such significant associations were not indicated when comparing body mass index, medical diagnoses, and whether they had undergone heart surgery., Conclusions: Adolescents with CHD practice health-promoting behavior similar to that of their counterparts without CHD. Health-promotion counseling for adolescents with CHD should be encouraged to improve lifestyle habits, especially to ensure that they engage in adequate and vigorous exercise and practice good dental hygiene.

A laser-based probe for the nonintrusive measurement of velocity gradient and vorticity was demonstrated in turbulent boundary layers. Unlike most other optical methods, the current technique provides an estimate of the velocity gradient, without having to first measure velocity at multiple points. The measurement principle is based on the heterodyne of coherent light scattered from two adjacent particles. The beat frequency of the heterodyne is directly proportional to the velocity gradient. The probe is assembled from commercially available, inexpensive optical components. A laser Doppler velocimeter (LDV) processor is used to analyze the heterodyne signal. A component of vorticity is obtained by using two appropriately aligned velocity gradient probes. The optical probes developed were used in turbulent boundary layers to measure local, time-frozen velocity gradients partial differential u / partial differential y, partial differential v / partial differential x, and partial differential v / partial differential y, as well as the spanwise vorticity. The measurements were compared to those inferred from LDV measurements in the same facility and to data available in the literature.

Background/aims: Drug metabolizing enzymes may be related to drug-induced liver injury (DILI). Manganese superoxide dismutase (MnSOD), NAD(P)H:quinone oxidoreductase (NQO1), and glutathione S-transferase (GST) are important drug metabolizing enzymes. We aimed to elucidate the relationship between genetic polymorphisms of these enzymes and the susceptibility to DILI., Methods: A total of 115 patients with DILI and 115 drug-, sex-, and age-matched controls were enrolled. Their genetic polymorphisms of MnSOD, NQO1, GSTM1, and GSTT1 were assayed., Results: Sixty-three (54.8%) of DILI patients were incriminated to anti-tuberculosis drugs. Subjects with a mutant C allele (T/C or C/C genotype) of MnSOD had a higher risk of DILI than those with MnSOD T/T genotype, both in overall drugs studied (adjusted OR: 2.44, 95% C.I.: 1.38-4.30, P=0.002), and in sub-category of anti-tuberculosis drugs (adjusted OR: 2.47, 95% C.I.: 1.13-5.39, P=0.02). In addition, subjects carrying GSTM1 null genotype had increased risk of anti-tuberculosis DILI (adjusted OR: 2.23, 95% C.I.: 1.07-4.67, P=0.03)., Conclusions: The MnSOD mutant C allele may increase the susceptibility to DILI, and GSTM1 null genotype may be related to anti-tuberculosis drug-induced hepatotoxicity. Determination of the MnSOD and GSTM1 genotypes may help identify patients at high risk for DILI.

Background: Polymerase chain reaction (PCR) is a molecular biology technique which can detect Mycobacterium tuberculosis (TB) genome in pleural fluid; however, the results are variable., Methods: Two hundred and twelve pleural fluid specimens suspected to be possibly associated with tuberculosis with negative acid-fast smears were sent to our laboratory to test for the presence of M. tuberculosis DNA using nested PCR, the target for the amplification being a segment of IS6110 in the genome of M. tuberculosis. The final diagnosis of TB pleurisy was based on combining clinical judgment with radiologic findings, microbiologic tests, and the histopathologic findings. Forty-nine patients were excluded due to incomplete or inconsistent clinical information., Results: Of 163 patients enrolled, PCR was positive in 23 (43.4%) of 53 patients with TB pleurisy and 5 (4.5%) of 110 patients with non-TB pleurisy, with a sensitivity and specificity of 43.4% and 95.5%, respectively. Positive culture of pleural fluid was found in 15 (28.3%) of the TB pleurisy group and none in the non-TB group. Fifteen (55.6%) of 27 with pleural biopsy demonstrated chronic granulomatous inflammation with or without caseous necrosis. Of these 27 patients, PCR was positive in 12 (44.4%). A higher proportion (70.4%) of patients with TB pleurisy was diagnosed when PCR was combined with biopsy results., Conclusion: These data indicate that PCR alone has limited value in diagnosis of TB pleurisy with negative smear. However, when used in combination with pleural biopsy, it can be used to increase early detection of TB pleurisy in such patients.

A myofibril-bound serine proteinase (MBSP) was highly purified from the skeletal muscle of crucian carp (Carasius auratus) by acidic treatment of myofibril solution and chromatographies on Q-Sepharose and benzamidine-Sepharose 6B. MBSP revealed a main protein band of approximately 28 kDa on SDS-polyacrylamide gel electrophoresis (PAGE) and was particularly inhibited by serine proteinase inhibitors. Substrate-specificity analysis revealed that the enzyme specifically cleaved at the carboxyl side of arginine and lysine residues, suggesting the characteristics of a trypsin-type serine proteinase. MBSP gene was cloned on the basis of the N-terminal sequence and the conserved active site peptide of serine proteinases together with 5'-rapid amplification of cDNA ends (5'-RACE) and 3'-RACE. The coding region gave an amino acid sequence of 242 residues including the initiation methionine and a signal peptide of 20 residues. Amino acid residues of His60, Asp106, and Ser196 consisting of the catalytic triad of serine proteinases were conserved in the sequence. Crucian carp MBSP shared relatively high identities with other serine proteinases, especially in well-conserved regions.

Background/purpose: To explore the etiology, age and gender distribution, complications, and prognosis of recurrent pediatric pancreatitis., Methods: Between 1993 and 2005, 92 children were hospitalized at the National Taiwan University Hospital with pancreatitis. Only 25 diagnosed with recurrent pancreatitis, based on two or more episodes of pancreatitis, elevated serum amylase and/or lipase levels > or = 3 times the upper limit of normal, radiographic evidence, and clinical symptoms, were enrolled., Results: A total of 85 episodes of pancreatitis in 25 patients (16 girls, 9 boys; mean age, 9.5 +/- 4.4 years; 3.4 +/- 1.9 episodes per person) were documented. The recurrence rate of pediatric pancreatitis was 27.2%. Recurrent pancreatitis was associated with pancreaticobiliary structural anomalies (n = 7), biliary stones or sludge (n = 4), hyperlipidemia (n = 3), pseudopapillary tumor of the pancreas (n = 2), trauma (n = 2), hypoxic encephalopathy with recurrent bacteremia and sepsis (n = 1), and idiopathic (n = 6). The age and gender distribution according to etiologies were not different (p = 0.301 for age, p = 0.137 for gender). Complications included cholangitis or cholestasis (16%), pancreatic necrosis (16%), pseudocyst formation (12%), shock (8%), hemorrhagic pancreatitis (4%), and diabetes mellitus (4%). No patient died of recurrent pancreatitis. Long-term morbidity after recurrent pancreatitis presented as gout, diabetes mellitus, non-alcoholic steatohepatitis, and chronic pancreatitis., Conclusion: For children who suffer from recurrent pancreatitis, pancreaticobiliary structural anomalies should be considered first.

Fungal infections have been increasing and life-threatening in recent years, but host immune responses, especially the humoral immunity, to fungi have not been fully understood. In the present study, we report that natural antibodies from unimmunized mice bind to Candida albicans. We established a monoclonal natural antibody, 3B4, which recognized a surface antigen located at germ tubes of C. albicans. The 3B4 antibody protected mice from C. albicans-induced death in passive immunization, by mechanisms involving suppressing germ tube formation and modulating phagocytosis. Interestingly, 3B4 also bound to a self-antigen keratin. To further study the generation and anti-C. albicans activities of natural antibodies in vivo, we constructed a mu chain transgenic mouse (TgV(H)3B4) using the V(H) gene from 3B4. TgV(H)3B4 had elevated serum anti-keratin/C. albicans IgM, and were resistant to C. albicans infections. Analyses of B cell development showed that in TgV(H)3B4, B cells secreting the anti-keratin/C. albicans antibodies were enriched in the B1 B cell compartment. Our findings reveal an important role of keratin-reactive natural antibodies in anti-C. albicans immune responses, and suggest that keratin may function in selecting B cells into the B1 B cell compartment, where natural antibodies are made to fight fungal infections.

There has been a decline in the incidence of acute rheumatic fever in recent decades in developed countries and in Taiwan. Sydenham's chorea, a major manifestation of rheumatic fever, was the most common cause of chorea in children in the past. But the incidence of Sydenham's chorea has declined in recent years in concert with the decline in rheumatic fever. Sydenham's chorea is usually bilateral and female predominant. Hemichorea is rare. We report on a 10-year-old boy who presented with progressive right side involuntary movements, an apical systolic murmur, prolonged PR interval, and elevated antistreptolysin O titer, who was diagnosed with acute rheumatic fever.

The spontaneous rupture of a Silastic catheter is a rare occurrence. We describe our experience of managing a tiny premature infant with embolization of a Silastic percutaneous central venous catheter (PCVC) and discuss the possible mechanisms of the embolization. A 28-week, 980 g, preterm male infant received a Silastic PCVC (Epicutaneo Cava Catheter, Vygon, Germany) for parenteral nutritional support at 4 days of age. The catheter was introduced percutaneously and advanced without difficulty through the right antecubital vein, and was subsequently withdrawn 2 cm following confirmation of tip position using radiography. A following chest radiograph, taken 15 hours later, showed rupture of the catheter, and an echocardiogram revealed a piece of the catheter had lodged between the right atrium and the right ventricle. The dislodged fragment of the catheter was retrieved successfully using a snare catheter (Microvena, White Bear Lake, Minn) by a pediatric cardiologist without complications. We want to stress that clinicians should be aware that rupture of the catheter is rare and can also occur asymptomatically and that an embolized fragment can be safely removed without extensive surgical manipulation, even in a tiny premature infant.

Background: Protein-losing enteropathy (PLE) is a serious complication of a Fontan operation and has a very high mortality rate. The purpose of this study was to investigate the incidence, clinical manifestations, diagnostic approaches, laboratory findings, therapeutic modalities and outcome of patients with PLE at our institution., Methods: The diagnosis of PLE was based on clinical manifestations and laboratory studies. We reviewed medical records of patients who received a Fontan operation at our hospital form July 1985 to October 2005., Results: A total 101 patients underwent various modifications of the Fontan procedure during this period. Nine of the 75 patients (12%) who survived 30 days after surgery developed PLE, including 4 boys and 5 girls. The median time interval between the Fontan operation and onset of PLE was 3.7 years (range 1.2 to 9.7 years). Laboratory examination showed low serum albumin levels and increased fecal alpha-1-antitrypsin excretion. Lymphangiectasia was found on intestinal biopsy. Six patients had cardiac catheterization after development of PLE which demonstrated an elevated mean right atrium pressure (22.5 +/- 6.4 mmHg, range 16 to 33 mmHg) and mean pulmonary artery pressure (22.3 +/- 6.4 mmHg, range 16 to 33 mmHg). Treatment included diet modification, albumin infusion, diuretics, inotropes, corticosteroids, heparin, and surgery. Four patients received medical treatment only. Two of these patients died due to sepsis and heart failure and 2 survived with partial relief of PLE. The remaining five received surgery for PLE after medical treatment failure. Three of them died after the operation and the two survivors were free of PLE, but one died of ventricular tachycardia 8 years later. The overall mortality rate was 67% (6/9)., Conclusions: The current treatment for PLE is associated with a very high mortality rate. Further investigation is needed to determine the exact mechanism of the disease and to develop new therapeutic approaches.

Mononuclear cells (MNCs) isolated from peripheral blood by density gradient centrifugation were plated on human fibronectin-coated culture plates and cultured in EGM-2 medium. Attached spindle-shaped cells, reported as endothelial progenitor cells (EPCs) by some investigators, had elongated from adherent round cells, but had not proliferated from a small number of cells as supposed previously. The growth curve of the primary EPCs showed that the cells had little proliferative capacity. Flow cytometry analysis showed that the cells could express some of the endothelial lineage markers, while they could also express CD14, which is considered a marker of monocyte/macrophage lineages throughout culture. In endothelial function assays, the cells demonstrated a lower level of expression of eNOS than mature endothelial cells in the reverse transcription-polymerase chain reaction and did not show an ability to develop tube-like structures in angiogenesis assay in vitro. In this study, we identified the monocytoid function of EPCs by the combined Dil-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and Indian ink uptake tests. All the cells were double positive for Dil-Ac-LDL and Indian ink uptake at days 4, 14 and 28 of culture, which means the EPCs maintained monocytoid function throughout the culture. Therefore, although adult EPCs from peripheral MNCs have some endothelial lineage properties, they maintain typical monocytic function and have little proliferative capacity.

At a medical center in Taiwan, all workers were examined by chest radiography, to determine the prevalence of pulmonary tuberculosis. The prevalence of tuberculosis among all hospital workers was 0.12%, that among nurses was 0.35%, and that among externally contracted cleaners was 0.57%. All of the Mycobacterium tuberculosis isolates recovered from 2 nurses and from a patient with pulmonary tuberculosis were the Beijing strain, but the strains had different serotypes.

Unlabelled: Asplenia syndrome is one of the heterotaxy syndromes, which many familial and animal studies suggest are caused by the loss of adequate genetic control of normal left-right asymmetry development. Moreover, there has not been any environmental factor documented to cause these syndromes. Asplenia syndrome occurring in a pair of monozygotic twins is reported. In view of the negative family history, a new germline mutant gene might be the aetiology of our patients. Both twins are associated with some degree of discordant complex heart defects within the context of a high degree of "mirror-image" of the unpaired thoracoabdominal organs., Conclusion: This report implies that sporadic asplenia syndrome might associate with new mutations and further genetic study may be indicated. These monozygotic twins' discordant phenotypes imply that some unidentified factors play an important role in their ultimate development of the same genetically determined abnormalities.

Coarctation of the aorta commonly presents in infancy as congestive heart failure, or later in childhood as hypertension or as a heart murmur. However, we experienced a unique infant case of isolated coarctation presenting with acute decompensation of a dilated cardiomyopathy, which recovered completely 8 months postoperatively. Our report highlights the previously unreported presentation of coarctation in infancy as a dilated cardiomyopathy. It also implies that before we label any patient presenting with a dilated cardiomyopathy as an idiopathic cardiomyopathy, we must exclude all possible specific causes of myocardial dysfunction because many such specific cardiomyopathies are curable and very rewarding, just like our patient., (Copyright 2006 S. Karger AG, Basel)

Background: For orientals, titrating doses of docetaxel (60-66 mg/m(2)) have shown equal effectiveness and fewer side effects as a second-line chemotherapy for patients with advanced non-small cell lung cancer (NSCLC). Under such doses, there were no comparative data between classic tri-weekly and Days 1 and 8 weekly schedules., Methods: This Phase II randomized prospective study was designed to compare the toxicity profile, efficacy and quality-of-life (QOL) between these two schedules of docetaxel in the treatment of previously treated patients with advanced NSCLC. Fifty patients were randomized to docetaxel arm A (66 mg/m(2) Day 1) and B (33 mg/m(2) Days 1 and 8) given every 3 weeks., Results: The overall response rates (ORRs) were 12 and 24% in arm A and B, respectively (P = 0.46), and disease control rates were 52 and 48%. The median time-to-progression (TTP) was 11.3 and 12.7 weeks and median survivals were 33.4 and 27.6 weeks, respectively. Both arms have same 1 year (36%) and 2 year survivals (12%). Arm A had significantly higher neutropenia but less compromised QOL. In this study, the response of second-line chemotherapy was significantly better in the group that was response to front-line chemotherapy (P = 0.032)., Conclusions: While Days 1 and 8 weekly docetaxel schedules show higher ORR and less hematological toxicity, there is no advantage to tri-week schedule in terms of TTP and survival, but more compromised QOL.

Full-length chicken interleukin-2 (ChIL-2) protein was successfully expressed using the recombinant baculovirus/Sf9 insect cell system. The expressed protein was soluble and reached approximately 12 microg/ml. Similarly to native ChIL-2, baculovirus expressed ChIL-2 revealed two main bands corresponding to molecular masses of 22 and 20 kD as detected by SDS-PAGE and Western blot. Treatment of the expressed protein with N-endoglycosidase F for 2 h caused the complete disappearance of the 22 kD band, while the 20 kD band (which is close to the molecular weight predicted from the cloned cDNA sequence) remained unchanged. Together with results on native ChIL-2, it can be concluded that ChIL-2 is an N-glycosylated protein.

Purpose: To describe ischemic retinopathy in a patient with tetralogy of Fallot., Design: Interventional case report., Testing: Clinical and imaging evaluation., Main Outcome Measures: Clinical, imaging, and laboratory findings in a patient with tetralogy of Fallot., Results: A 20-year-old female patient with tetralogy of Fallot had progressive visual loss of 3 weeks duration. Bilateral examination revealed dilated, tortuous, conjunctival vessels; prominent anterior chamber reaction; iris neovascularization; posterior synechia; retinal vascular tortuosity in both eyes; and inferior exudative retinal detachment. Fluorescein angiography revealed delayed retinal and choroidal filling. The working diagnosis was ischemic retinopathy with uveitis. The patient was treated for 6 months with a high-dose oral corticosteroid combined with a topical corticosteroid, a topical mydriatic, and panretinal photocoagulation. Conjunctival, vascular congestion subsided with a decrease in anterior chamber reaction. The inferior, exudative retinal detachment resolved, and vision was restored., Conclusions: Retinal ischemic syndrome combined with uveitis can develop in patients with tetralogy of Fallot. Treatment can restore vision in such patients.

A trypsin-type serine proteinase (WSP) was purified previously from the sarcoplasmic fraction of skeletal muscle of white croaker (Argyrosomus argentatus) by Yanagihara et al. ((1991) Nippon Suisan Gakaishi, 57, 133-142). However, further research on WSP was not carried out. In the present study, we determined the N-terminal amino acid sequence of this enzyme (27 amino acid residues), which revealed relatively high identity in the conserved region to other trypsin-type serine proteinases. Degradation action of WSP on neuropeptides is also reported in this manuscript. The results show that WSP only cleaves at the carboxyl side of Arg or Lys residue of the peptides, especially between dibasic amino acid residues such as Arg-Arg and Arg-Lys.

Vein of Galen aneurysmal malformation (VGAM) often leads to death in the neonatal period, mainly due to congestive heart failure. Chronic and excessive pulmonary flow in utero and postnatally is attributed to large VGAM and right ventricular overload. We report a male neonate with VGAM complicated by severe heart failure and persistent pulmonary hypertension. Endovascular coil embolization of VGAM was performed, resulting in improvement of congestive heart failure; however, severe persistent pulmonary hypertension led to death 2 days after the embolization. Postmortem examination showed marked right and left ventricular hypertrophy and impressive muscular thickening of intra-alveolar arterioles. Neonatal embolization seems to be beneficial only in babies without suprasystemic pulmonary hypertension. Therefore, early delivery and repair of VGAM should be considered before the onset of persistent pulmonary hypertension.

In order to develop a rapid method which can check Campylobacter jejuni in animal and poultry foods nicely, an immunomagnetic capture-fluorescent PCR (IMC-FPCR) method was established in this paper. The reported method involves isolation of the target pathogen by immunocapture prior to the fluorescent PCR step, therefore the immunomagnetic-beads for Campylobacter were developed, and two groups of primer/probe, which targeted for the species special sequence of flaA gene and hipO gene for Campylobacter jejuni were designed. The immunomagnetic capture-fluorescent PCR assay amplification of the hipO gene and flaA gene for detection of Campylobacter jejuni was firstly reported in this paper. Result indicated that IMC-FPCR method permits direct detection of the pathogen without an enrichment step and can be performed in approximately 24 h. The assay results are positive for all of the isolates of Campylobacter jejuni (3 isolates, including type strain ATCC 33560 and ATCC8341) with a detection limit of approximately 10 cfu/mL, are negative for Campylobacter coli and several other bacteria. IMC-FPCR assay provide not only a rapid, sensitive method for quantitative detection of Campylobacter jejuni, but also an important method for detecting of Campylobacter jejuni of viable but non-culturerable (VNC) state.

Unlabelled: Two infants developed hoarseness unexpectedly the day after transcatheter coil closure of a slender patent ductus arteriosus (PDA). The pathogenesis of this complication appears to be similar to that of the classic cardiovocal syndrome. During the intervention, the inappropriately implanted coil might have distorted the slender PDA, thereby causing angulation of the pliable PDA itself and precipitating impingement on the left recurrent laryngeal nerve. Fortunately, both infants recovered spontaneously from the hoarseness within several weeks. At present, the definite underlying neuropathology of this complication is unknown as we have not yet confirmed recovery of the left vocal cord movement by follow-up fibreoptic bronchoscopy., Conclusion: Iatrogenic cardiovocal syndrome could occur in infants after transcatheter coil closure of a slender PDA, using the currently popular 0.038-inch coil. A coil with a smaller diameter might prevent the occurrence of this syndrome.

Objective: To define the clinical characteristics and clinical course of hospital-acquired severe acute respiratory syndrome (SARS)., Patients and Methods: This 8-month prospective study of 14 patients with hospital-acquired SARS in Taipei, Taiwan, was conducted from April through December 2003., Results: The most common presenting symptoms in our 14 patients with hospital-acquired SARS were fever, dyspnea, dizziness, malaise, diarrhea, dry cough, muscle pain, and chills. Lymphopenia and elevated serum levels of lactate dehydrogenase (LDH) and C-reactive protein (CRP) were the most common Initial laboratory findings. Initial chest radiographs revealed various pattern abnormalities and normal results. Five of the 14 patients required mechanical ventilation. The need for mechanical ventilation was associated with bilateral lung involvement on the initial chest radiograph and higher peak levels of LDH and CRP. Clinical severity of disease varied from mild to severe. At 8 months after disease onset, patients with mild or moderate SARS had normal findings or only focal fibrosis on chest high-resolution computed tomography. However, bilateral fibrotic changes remained in the 4 patients who had recovered from severe SARS, 1 of whom had mild restrictive ventilatory impairment. One patient with severe SARS died; she was elderly and had other comorbidities. Five additional patients had reduced diffusing capacity., Conclusion: The clinical picture of our patients presenting with hospital-acquired SARS revealed atypical pneumonia associated with lymphopenia, elevated serum levels of LDH, rapid clinical deterioration, and lack of response to empirical antibiotic therapy. Substantially elevated levels of LDH and CRP correlated with severe illness requiring mechanical ventilatory support. In those receiving mechanical ventilation, pulmonary function was only mildly reduced at 6 to 8 months after acute illness, consistent with the natural history of acute respiratory distress syndrome due to other causes.

Glycogen storage disease type II (GSD-II), also known as Pompe disease, is a rare autosomial recessive disease due to deficiency of lysosomal acid alpha-glucosidase (GAA). The infantile-onset form is the most severe, and most patients present with hypotonia and cardiomyopathy in early infancy. We report on a typical case of Pompe disease in a patient who died at 8 months of age due to aspiration pneumonia and hypertrophic cardiomyopathy. Genetic studies showed deficient GAA activity and mutation of the GAA gene with Gly615Arg (exon 13, G1845A). On autopsy, glycogen had markedly accumulated in the liver, myocardium and skeletal muscle. The neurons of the anterior horn of the spinal cord and medulla were also involved, but the cortex was spared. These neurological-histologic findings may explain the clinical features of poor motor function, decreased deep tendon reflexes and lack of mental retardation.

All-trans retinoic acid (ATRA) affects cell proliferation, differentiation and apoptosis through its receptors, RARs and RXRs. Besides these, other receptors such as orphan receptor TR3, are also involved in the regulatory process of ATRA. However, how different receptors function in response to ATRA is still largely unknown. In the present study, we found that formation of TR3/RXRalpha heterodimers in the nucleus and their subsequent translocation into the cytoplasm, in association with regulation of apoptosis-related proteins Bcl-2, Bcl-xl and Bax, was critical for apoptosis induction by ATRA in breast cancer cells MCF-7. When such translocation was blocked by Leptomycin B (LMB), ATRA-induced apoptosis was consequently abolished. However, in ATRA-induced gastric cancer cells MGC80-3, RXRalpha heterodimerised with RARalpha but not with TR3, and remained in the nucleus exerting its effect on cell cycle regulation. When transfected with antisense-RARalpha, MGC80-3 cells changed from ATRA-sensitive to ATRA-resistant and most cells were arrested in the S phase, implying the importance of RARalpha in cell cycle regulation. Furthermore, we demonstrated that the effects of ATRA depend on the relative levels of TR3, RARalpha and RXRalpha expression in cancer cells. In ATRA-induced MCF-7 cells, highly expressed TR3 favours the formation of TR3/RXRalpha and promotes the TR3/RXRalpha signalling pathway causing apoptosis; while in ATRA-induced MGC80-3 cells, high expression of RARalpha favours the formation of RARalpha/RXRalpha and promotes the RXRalpha/RARalpha signalling pathway in mediating cell cycle regulation. In conclusion, these results reveal the novel mechanism that cellular expression and location of protein is associated with diverse signalling transduction pathways and the resultant physiological process.

Background and Purpose: Discrete subaortic stenosis (DSS) is considered an acquired cardiac defect. However, its clinical course and pathogenesis have not been well defined. This study used echocardiography to investigate the natural history of DSS and identify the possible anatomic abnormalities leading to its development., Methods: We reviewed the medical records of 12 children with a diagnosis of DSS between 1988 and 2002. Data on the age at initial diagnosis of DSS was collected and the sequential changes of left ventricular outflow tract obstruction (LVOTO) and aortic regurgitation (AR) were analyzed by serial echocardiographic studies. Patients were divided into 2 subgroups according to the latest or presurgical (in patients with operations for DSS) Doppler-derived peak instantaneous left ventricular outflow tract gradient (deltaP; cut-off point: deltaP 50 mm Hg, our institutional criterion of operation for DSS). The indexed mitral valve (MV)-aortic valve (AV) distance, aortoseptal angle (ASA), indexed DSS-AV distance, and whether the anterior MV leaflet was involved were also determined. The DSS patients were compared with an age- and lesion-matched control group (12 patients)., Results: The mean age at initial diagnosis of DSS was 4.9 +/- 3.7 years. Nine children had disease characterized by milder LVOTO (latest deltaP < 50 mm Hg) and slower progression of LVOTO (mean increase of deltaP, 7.1 +/- 4.4 mm Hg/year) and AR ( 50 mm Hg) and more rapid progression of LVOTO (mean increase of deltaP, 27.8 +/- 8.4 mm Hg/year) and AR (>/= grade II). Compared with the control group, the study group had a significantly longer indexed MV-AV distance and a steeper ASA. Compared with the patients with milder disease, patients with more severe disease had a significantly shorter indexed DSS-AV distance and more frequent involvement of the anterior MV leaflet., Conclusions: These data indicate that DSS may present with 2 distinct clinical courses. A shorter indexed DSS-AV distance and the involvement of the anterior MV leaflet might be predictive of more rapidly progressive DSS. Longer indexed MV-AV distance and steeper ASA might be useful to evaluate primary cardiac defects and thus to identify patients who are at risk for developing DSS.

The KAI1 gene is identified as a tumor metastasis suppressor gene in many types of cancer. We examined KAI1 gene and its protein KAI1/CD82 expression by in situ hybridization and immunohistochemical analysis, and found that KAI1 mRNA and protein expression were inversely correlated with lymph node and distant metastasis in digestive tract carcinomas, but not with age and gender of the patient, or with tumor differentiation. Moreover, KAI1/CD82 protein expression positively reflected the survival outcome of patients. Western blot analysis showed that VP-16 increased KAI1/CD82 protein expression obviously in various cancer cell lines, especially in those that were highly metastatic. This increased KAI1/CD82 expression was associated with its translocation from the cytomembrane to the nucleus, in which it interacted with nuclear p53 protein, forming a strong complex, observed by confocal microscopy and co-immunoprecipitation, respectively. In nude mice, after feeding with VP-16, the number of tumors metastasized from spleen to liver was obviously reduced, and KAI1/CD82 protein expression became stronger in those metastatic tumors. Accordingly, this demonstrated that KAI1 might be used as an indicator for predicting the clinical outcome, and VP-16 may be clinically considered as a promising candidate for anti-metastasis with regard to its potential to upregulate KAI1 expression.

Aim: To investigate and determine the mechanism and signal pathway of tetradecanoylphorbol-1, 3-acetate (TPA) in degradation of RXRalpha., Methods: Gastric cancer cell line, BGC-823 was used in the experiments. The expression level of RXRalpha protein was detected by Western blot. Nuclear and cytoplasmic protein fractions were prepared through lysis of cell and centrifugation. Localization and translocation of RXRalpha were observed under laser-scanning confocal microscope through labeling specific anti-RXRalpha antibody and corresponding immunofluorescent antibody as secondary antibody. Different inhibitors were used as required., Results: In BGC-823 cells, RXRalpha was expressed in the nucleus. When cells were treated with TPA, expression of RXRalpha was repressed in a time-dependent and TPA-concentration-dependent manner. Meanwhile, translocation of RXRalpha from the nucleus to the cytoplasm occurred, also in a time-dependent manner. When cells were pre-incubated with proteasome inhibitor MG132 for 3 hrs, followed by TPA for another 12 hrs, TPA-induced RXRalpha degradation was inhibited. Further observation of RXRalpha translocation in the presence of MG132 showed that MG-132 could block TPA-induced RXRalpha redistribution. Conversely, when RXRalpha translocation was inhibited by LMB, an inhibitor for blocking protein export from the nucleus, TPA could not repress expression of RXRalpha., Conclusion: TPA could induce the degradation of RXRalpha protein in BGC-823 cells, and this degradation is time- and TPA-concentration-dependent. Furthermore, the degradation of RXRalpha by TPA is via a proteasome pathway and associated with RXRalphatranslocation from the nucleus to the cytoplasm.

Background: Outbreak of severe acute respiratory syndrome (SARS) in Taipei has been associated with Taiwanese back from Guangdong, China. We report 4 probable SARS cases with different severity and propose optimal treatment., Methods: Four probable SARS cases were enrolled. Two cases were due to outbreak of SARS in our hospital and two cases were transferred from other hospitals. All patients received standard treatment: ribavirin 1000 mg orally daily for 10 days, Levofloxacin 500 mg orally daily for 7 days, and intravenous immunoglobulin (IVIG) 1 g/kg/day for 2 day after the onset of symptoms. If severe hypoxia (PaO2/FiO2 < 200) occurred, protective strategy of mechanical ventilation and methylprednisolone 2 mg/kg/day were given. The clinical pictures and treatment outcome were discussed., Results: Fever, dyspnea, diarrhea, malaise, dizziness and dry cough were initially more common symptoms. Initially chest patterns included focal consolidation, interstitial infiltration or normal. Common laboratory findings were lymphopenia, and elevated serum levels of lactate dehydrogenase and C-reactive protein. No mortality was found., Conclusions: Highly alert and stringent infection control of SARS cases are required. Otherwise, SARS easily induces hospital-acquired first then community-acquired infection. Initial presentation of radiographic patterns includes normal, interstitial or airspace shadowing. Fever and lymphopenia are occasionally followed by rapidly progressive respiratory compromise. The standard treatment might be beneficial for decreasing the mortality rate.

Most cases with antituberculosis drug-induced hepatitis have been attributed to isoniazid. Isoniazid is metabolized by hepatic N-acetyltransferase (NAT) and cytochrome P450 2E1 (CYP2E1) to form hepatotoxins. However, the role of CYP2E1 in this hepatotoxicity has not yet been reported. The aim of this study was to evaluate whether the polymorphism of the CYP2E1 gene is associated with antituberculosis drug-induced hepatitis. A total of 318 tuberculosis patients who received antituberculosis treatment were followed prospectively. Their CYP2E1 and NAT2 genotypes were determined using a polymerase chain reaction with restriction fragment length polymorphism method. Twenty-one healthy volunteers were recruited for CYP2E1 phenotype study using a chlorzoxazone test. Forty-nine (15.4%) patients were diagnosed to have drug-induced hepatotoxicity. Patients with homozygous wild genotype CYP2E1 c1/c1 had a higher risk of hepatotoxicity (20.0%; odds ratio [OR], 2.52) than those with mutant allele c2 (CYP2E1 c1/c2 or c2/c2, 9.0%, P =.009). If CYP2E1 c1/c2 or c2/c2 genotype combined with rapid acetylator status was regarded as the reference group, the risk of hepatotoxicity increased from 3.94 for CYP2E1 c1/c1 with rapid acetylator status to 7.43 for CYP2E1 c1/c1 with slow acetylator status. After adjustment for acetylator status and age, the CYP2E1 c1/c1 genotype remained an independent risk factor for hepatotoxicity (OR, 2.38; P =.017). Furthermore, under the administration of isoniazid, the volunteers with CYP2E1 c1/c1 genotype had higher CYP2E1 activity than those with other genotypes had and, hence, might produce more hepatotoxins. In conclusion, CYP 2E1 genetic polymorphism may be associated with susceptibility to antituberculosis drug-induced hepatitis.

Aim: JNK cascade plays an important role in cell proliferation, differentiation and apoptosis. However, the exact function of JNK cascade for apoptosis induction remains largely unknown. In this study, the role of JNK activation stimulated by TPA in the process of apoptosis induction and its signaling transduction pathway in gastric cancer cells were investigated and determined., Methods: Expressions of mRNA and protein were detected by Northern blot and Western blot. Transcription activity was measured by transient transfection and CAT assay. Apoptotic cells were displayed through staining the nucleus with DAPI and were observed under fluorescence microscope. The apoptotic index was determined by counting 1000 cells randomly., Results: JNK protein was stimulated rapidly by TPA, and reached its highest peak within 3 hr, then decreased in a time-dependent manner, but the expression level of JNK protein induced by TPA was always keeping higher than that in untreated cells. Similar pattern was seen in c-jun mRNA level induced by TPA. TPA significantly activated the transcriptional activity of activator protein-1 with a TPA-dose-dependent manner. Furthermore, activation of JNK was mediated through PKC pathway. Treatment of cells with PKC specific inhibitor, Wortmannin, led to repression of JNK even in the presence of TPA. More importantly, all these effects were associated with induction of apoptosis in gastric cancer cells. TPA inducted apoptosis obviously in gastric cancer cells. The apoptotic cells became smaller and rounded, and their nuclei became condensation and fragmentation with brightly stained chromatin. However, suppression of JNK by PKC specific inhibitor, Wortmannin, resulted in the decrease of apoptosis induced by TPA in a time-dependent manner, apoptotic index dramatically decreased from 32.56 % to 8.71 %., Conclusion: TPA stimulates JNK cascade, including up-regulation of JNK protein expression level and c-jun mRNA expression level, and activation of activator protein-1 transcriptional activity. Activation of JNK is mediated through PKC pathway, which has an association with induction of apoptosis by TPA. Thus, activation of JNK via PKC pathway may represent one of important mechanisms for TPA to induce apoptosis in gastric cancer cells.

To date, the diagnosis of tuberculosis has not improved significantly and still relies heavily on staining and culture of sputum or other clinical specimens which were developed more than 100 years ago. Staining does not differentiate tuberculosis from other mycobacterial infections, and culture requires at least 4 to 8 weeks. These are the major problems faced by tuberculosis control programs. In response to this demand, new rapid diagnostic methods are urgently sought. In recent years, much hope has been laid on the development of molecular techniques in the routine tuberculosis laboratory. This review concentrates on 4 techniques that are increasingly used in clinical laboratories: polymerase chain reaction to detect mycobacterial DNA in clinical specimens, nucleic acid probes to identify culture, restriction fragment length polymorphism analysis to compare strains for epidemiologic purposes, and genetic-base susceptibility testing methods for rapid detection of drug resistance. Finally, the increase in the use of clinically-useful molecular biological techniques that affect turnaround time, length of stay, and patient outcome, and reduce overall hospitalization costs will continue until universal standardization for molecular diagnostic procedures are provided. At present, conventional methods should not be replaced by novel methods until the latter are shown to be of equal or greater sensitivity, specificity, reliability, and user-friendliness. However, it is expected that the newly developed molecular techniques will complement our armamentarium of diagnostic tools in the detection of tuberculosis. It is also expected that clinical protocols based on molecular methods will increase the chances for cure by selecting the most appropriate treatment and improving the quality of life of tuberculosis patients.

Setting: Veterans General Hospital-Taipei, Taiwan., Objective: To assess the efficacy and safety of a fixed-dose combination (FDC) of Rifater (RFT)/Rifinah (RFN) in the treatment of newly diagnosed smear-positive pulmonary tuberculosis., Design: Patients were randomly assigned to two 6-month short-course chemotherapy regimens. One group of patients was treated with FDCs and another was given the four component drugs (INH, RMP, EMB and PZA) as separate formulations., Results: The 105 patients enrolled in the study were divided into two treatment groups. Fifty-one patients who had completed treatment without interruption, 26 in the FDC group and 25 in the separate regimen, were eligible for analysis at the end of 2 years. Among the patients with a drug susceptibility test result available, four in the FDC group had bacilli resistant to pyrazinamide. In the separate regimen group, two patients had bacilli resistant to ethambutol and six had bacilli resistant to pyrazinamide. The two regimens were of similar effectiveness with regard to sputum conversion, compliance and radiological improvement. No patient with FDC treatment developed gastointestinal symptoms, visual disturbance or peripheral neuropathy (P < 0.05). However, FDC treatment resulted in drug-induced fever in one patient. One patient (3.8%) in the FDC group relapsed 5 months after completing treatment., Conclusion: This study suggests that the two regimens had similar effectiveness in the treatment of smear-positive pulmonary tuberculosis. However, the fewer adverse drug events among those patients treated with the FDC regimen suggests that it has a better safety profile.

Nur77 is an orphan receptor. Although Nur77 affects cell proliferation and apoptosis through its capability of binding to a variety of response elements and regulating their transactivation activities, the intrinsic function of Nur77 is not yet fully understood; in particular, its regulation of apoptosis and proliferation has been characterized as cell type-dependent and agent context-dependent. In this study, Nur77 can be seen to regulate apoptosis via its expression and translocation, rather than its transactivation activity in gastric cancer cells. Nur77 was constitutively expressed in BGC-823 cells. The tetradecanoylphorbol-1,3-acetate (TPA) treatment not only resulted in up-regulation of the Nur77 mRNA level, but also led to translocation of Nur77 protein from the nucleus to the mitochondria, and caused the release of cytochrome c. This TPA-induced translocation of Nur77 was in association with the initiation of apoptosis in gastric cancer cells. Although all-trans retinoic acid (ATRA) could not induce apoptosis in BGC-823 cells due to failure of stimulating Nur77 translocation, expression of Nur77 in the nucleus was required for cell growth inhibition by ATRA. Transfection of antisense Nur77 receptor into BGC-823 cells resulted in resistance of cell growth against ATRA inhibition, and the cells were still arrested in the S phase. Furthermore, the action of Nur77 in TPA-induced apoptosis was mediated through a protein kinase C signaling pathway, while mitogen-activated protein kinase and phosphatidylinositol 3-kinase signaling pathways were responsible for the regulation of Nur77 mRNA expression. Taken together, the data revealed the dual functioning mechanisms of Nur77 in gastric cancer cells in response to TPA and ATRA.