Shiuh-Bin Fang

Abstract: In this study, we investigated the anti-inflammatory and reinforcing barrier effects of Lactobacillus casei rhamnosus (Lcr35) on Caco-2 intestinal epithelial cells (IECs) already exposed to Salmonella lipopolysaccharide (LPS). Using the transwell co-culture model, Salmonella LPS was apically added to polarized Caco-2 cells co-cultured with peripheral blood mononuclear cells (PBMCs) in the basolateral compartment. LPS-stimulated Caco-2 cells were incubated with Lcr35 for 1, 6, 24 or 48 hours. Apical inoculation of Lcr35 after 48 hours significantly inhibited the basolateral secretion of IL-8 in the Caco-2/PBMCs co-culture. The PCR analysis showed that Lcr35 significantly downregulated mRNA expression of monocyte chemoattractant protein 1 (MCP-1)(P<0.05), had a trend to decrease mRNA levels of interleukin-8 (IL-8)(P=0.05), but did not alter mRNA expression of transforming growth factor-beta1 (TGF-beta1 ) in LPS-stimulated Caco-2 cells at 48 hours after addition of Lcr35. Compared to non-LPS-pretreated controls, transepithelial electrical resistance (TEER) of the polarized Caco-2 cell monolayers pretreated with LPS for 48 hours was decreased by 9.9% (P<0.05). Additionally, compared to those cells only treated with LPS, apical co-incubation with Lcr35 showed a biphasic increased TEER levels by 12.1% (P<0.001), 5.7% (P<0.05), and 86.8% (P<0.001) in the Caco-2 cell monolayers than those without Lcr35 treatment after 1, 6, and 48 hours, respectively. In conclusion, Lcr35 can exert anti-inflammatory effects and ameliorate barrier dysfunction in the Salmonella LPS-pretreated inflamed intestinal epithelium in vitro.

Abstract: In this study, we developed an easy screening test that identifies Salmonella in 2 h after colony-print which is a procedure based on the transfer of surface cells of the colonies to a nitrocellulose membrane. Gold nanoparticles coated with the anti-Salmonella antibody were used to highlight the Salmonella spp. on the membrane to facilitate the selectivity. On Hektoen agar, 134 stool samples containing black or crystalloid colonies were identified using the proposed method. Without any equipment, such as microscope, the red dots corresponding Salmonella were observed. After colony-print test, 22 of the isolates were correctly identified as Salmonella to achieve 100% sensitivity. 111 samples were correctly identified as non-Salmonella spp., but one was incorrectly identified as Salmonella. The specificity is 99.1%. This method is simple, straightforward, inexpensive, and fast. It can be easily applied to the routine workload of clinical laboratories, and can be very useful when large amounts of fecal samples should be evaluated for rapid screening and diagnosis.

Abstract: Beneficial effects of probiotics in acute infectious diarrhoea in children are mainly seen in watery diarrhoea and viral gastroenteritis. Lactobacillus rhamnosus, one the most extensively studied probiotic strains, is effective in shortening courses of acute diarrhoea in children. However, the dose-dependent effect of Lactobacillus upon quantification of faecal rotavirus shedding in humans remains little known. Thus, an open-label randomized trial in 23 children with acute rotaviral gastroenteritis was undertaken by randomly allocating patients to receive one of the three regimens for 3 days: daily Lactobacillus rhamnosus 35 (Lcr35) with 0 CFU/day to six patients in the control group, 2 x 10(8) CFU/day to nine patients in the low-dose group, and 6 x 10(8) CFU/day to eight patients in the high-dose group. Faecal samples were collected before and after the 3-day regimen for measurements of rotavirus concentrations by ELISA. There was no statistically significant change in faecal rotavirus concentrations in either the control group (119.2 x 10(5) particles/ml vs. 23.7 x 10(5) particles/ml, p = 0.075) or the low-dose group (36.1 x 10(5) particles/ml vs. 73.5 x 10(5) particles/ml, p = 0.859). However, the high-dose group had a significant reduction of faecal rotavirus concentration (64.2 x 10(5) particles/ml vs. 9.0 x 10(5) particles/ml, p = 0.012). Without any exception, the faecal rotavirus concentrations of all eight patients in the high-dose Lcr35 group declined by 86% after 3 days when compared with those before Lcr35 administration. In conclusion, this is the first report to provide quantitative evidence of the dose-dependent effect of Lactobacillus rhamnosus, a minimal effective dose of 6 x 10(8) CFU for 3 days, upon the faecal rotavirus shedding in paediatric patients.

Abstract: This study developed a method of detecting bioparticles such as Salmonella that exist in the biological samples. The method employed a substrate with interlaced comb-like electrodes into which the mixtures of biological samples and antibody-coated gold nanoparticles were added. The alternative signals with appropriate frequency bands were then conducted into the comb-like electrodes to change the dielectrophoresis force. The gold-modified Salmonella can be adsorbed on the edges of the electrodes and isolated from various biological samples. The impedance of the adsorbed Salmonella on the edges of the electrodes was measured and comparison of the impedance between the electrodes with and without Salmonella can quantify the amount of the adsorbed Salmonella.

Abstract: In this study, we developed an easy and quick test capable of identifying specific bacteria in one hour. The protocol was established based on the measurement of bacteria quantity on a biochip with comb-like electrodes. Gold nanoparticles coated with anti-Salmonella antibody were used to enhance the dielectrophretic property of Salmonella spp. on the biochip to facilitate the sensitivity.

Abstract: This study aims at developing a 3D device for catching, separating, and transporting bio-particles based on dielectrophoresis (DEP). Target particles can be simultaneously caught and transported using the negative DEP method. In non-uniform electric fields, the levitation height or complex permittivity of certain particle may be different from that of another and this property can facilitate separation of particles. We have designed and constructed a 3D device consisting of two layers of electrodes separated by a channel formed by 50 microm thick photoresist. The electrodes can operate effectively with 10-15 V and 5-7 MHz to catch all particles in the channel, and can move particles after switching the electric field to 5-15 V and 500-1,000 KHz. Hence, particles experienced coupling force of two different directional twDEP forces, and tallied with our estimation to move along the coupling direction.

Abstract: BACKGROUND: Intracranial hemorrhage (ICH) in full-term infants is uncommon and is a rare cause of neonatal fever. This study was conducted to estimate the incidences of ICH and fever in a hospital-based population, and to determine if the clinical features of neonatal ICH with and without hyperthermia differ. METHODS: We selected 315 afebrile neonates who received cranial ultrasonography screening from 2003 to 2004 as the control group, and 153 neonates diagnosed with fever from 1998 to 2004 as the study group. During the same period, 28 full-term neonates with birth weights >2500g and ICH in the first week of life were enrolled retrospectively. They were divided into hyperthermia (n=11) and nonhyperthermia (n=17) groups. RESULTS: Three babies in the control group and 11 in the study group had ICH; the incidence of ICH in the hyperthermia group was significantly higher (3/315 vs. 11/153; p < 0.001). Compared with the nonhyperthermia group, the hyperthermia group had Less cyanosis (2/11 vs. 10/17; p = 0.04), Less lymphocyte predominance (33.7 vs. 80%; p = 0.032), higher neutrophil/lymphocyte ratio (1.9 vs. 0.3; p = 0.006), higher erythrocyte/leukocyte ratio (425 vs. 79.5; p = 0.05) in cerebrospinal fluid and an increased incidence of subarachnoid hemorrhage accompanied by intracerebral parenchymal hemorrhage (4/11 vs. 0/17; p = 0.016). CONCLUSION: The incidence of ICH was higher in febrile than in afebrile neonates. ICH presenting with hyperthermia might go unrecognized, because its other symptoms are subtle and a neutrophil predominance in the cerebrospinal fluid might result in a misdiagnosis of meningitis. ICH in the interior brain tended to manifest more commonly as hyperthermia than did ICH in the superficial brain. A cranial image examination should be considered in the evaluation of neonatal fever.

Abstract: BACKGROUND: This study analyzed the clinical presentation, postoperative morbidity and mortality and incidence of associated extrahepatic biliary atresia in children with biliary cysts in Taiwan. METHODS: We retrospectively reviewed the records of 158 pediatric patients with biliary cysts seen between June 1981 and July 2004, with follow-up ranging from 12 months to 22 years (mean, 11.2 +/- 6.1 years). Patients were divided into three groups: biliary atresia-associated biliary cyst (BABC, 21 patients), non-biliary atresia-associated choledochal cyst (NBACC) in infancy (37 patients), and late NBACC (> 1 year of age, 100 patients). RESULTS: BABC accounted for 36.2% of the infantile biliary cysts in this study. Extrahepatic cysts in late NBACC had a greater mean diameter than those in infantile NBACC and BABC (21.5 mm vs. 16.0 mm vs. 7.9 mm, p < 0.001). Cholangitis was the most serious complication within 3 months postoperatively in all three groups, resulting in four deaths (two in the infantile NBACC group and one each in the other two groups). Liver cirrhosis developed during long-term follow-up in nine of the 21 patients with BABC, four of whom died. Three of these nine patients underwent liver transplantation and remained well during follow-up. Chronic complications in NBACC occurred mainly in late IVa cases, with persistent intrahepatic dilatation developing in 12 of 35 patients and intrahepatic stones in five. Elevation of serum alanine aminotransferase (ALT) was found preoperatively in 85% of late NBACC and 35% of infantile NBACC cases. Postoperative normalization of ALT occurred after a mean of 152 +/- 23 days and 158 +/- 67 days in late NBACC and infantile NBACC, respectively. Higher ALT levels before operation were associated with a longer period until normalization. CONCLUSION: The possibility of BABC must be included in the differential diagnosis when a small extrahepatic cyst (< 8 mm in diameter) with prolonged jaundice is found in infancy. Postoperative follow-up is essential for patients with NBACC due to their frequently prolonged elevation of serum ALT and possibility of residual intrahepatic dilatation. Cholangitis was the major cause of death within 3 months postoperatively in this study.

Abstract: Osteolysis induced by ultra-high molecular weight polyethylene wear particles is the major cause of long-term failure of artificial joints. We examined the effects of wear particles on bioactivity by analysing the biophysical aspects of particle phagocytosis. We estimated the maximum number of internalized particles (the phagocytic capacity) for particles of various sizes and shapes. We demonstrated that elongated particles had a smaller phagocytic capacity than spherical particles of the same volume. A review of the literature showed that the ratio of particle concentration (number of particles/number of cells) to phagocytic capacity is critical for particle-induced biological responses. When this ratio was < 1, the biological response was approximately proportional to the ratio itself. When this ratio was > 1, limited changes in the biological response were observed. The saturation level of the phagocytic capacity for a particle population appears to reflect the degree of polyethylene particle-induced biological response.

Abstract: OBJECTIVE: To present the first reported case of early second trimester maternal hepatitis C virus (HCV) associated with fetal ascites, which was treated with fetal paracentesis, and resulted in a successful outcome of a term liveborn infant with anti-HCV seropositivity. CASE REPORT: A 26-year-old primigravida woman was diagnosed with acute HCV infection at 17 weeks of gestation. Ultrasound (US) at 23 weeks showed significant fetal as cites and echogenic bowel, and fetal viral infection was suspected. Maternal serum was positive for high HCV-RNA titers and cytomegalovirus (CMV) IgG. Amniocentesis, cordocentesis and therapeutic fetal paracentesis were performed at 23 weeks. Fetal karyotype was 46, XX. Cord blood showed anti-HCV positivity and HCV-RNA titer < 10. Amniotic fluid was anti-HCV and CMV IgG positive. US at 27 weeks showed complete resolution of fetal ascites. A healthy 2,976 g female baby was delivered at 37 weeks, with anti-HCV seropositivity, high HCV-RNA titers, CMV IgG positive, IgM negative and normal liver function tests at the 1-month follow-up. CONCLUSION: Second trimester perinatal HCV infection with possible CMV coinfection associated with fetal ascites is a rare event. Fetal therapy resulting in a successful outcome has not been reported. Prompt fetal therapy with paracentesis in this case led to the delivery of a healthy term liveborn baby with anti-HCV seropositivity.

Abstract: AIMS: The aim was to investigate conjugated and unconjugated hyperbilirubinaemia in association with urinary tract infection (UTI) in young infants. METHODS: Fifty infants aged <3 mths who developed prolonged jaundice among 2128 infants with UTI from 1984 to 2004 were enrolled retrospectively. They were divided into conjugated (n=22) and unconjugated (n=28) hyperbilirubinaemia groups and the clinical variables between the two were compared. RESULTS: Compared with the unconjugated group, the conjugated hyperbilirubinaemia group had statistically significantly lower haemoglobin (1.57 vs 1.80 micromol/L), higher aspartate aminotransferase (96 vs 32.5 U/L) and alanine aminotransferase (81.5 vs 16 U/L), were older on admission (48.0 vs 32.5 days), had a longer duration of jaundice before treatment (43.5 vs 30 days) and a higher incidence of E. coli infections (19/22 vs 15/28). The direct/total bilirubin ratio was linearly correlated with duration of jaundice before treatment (p=0.004). The most significant cut-off value for the duration of jaundice vis-à-vis the type of jaundice was 38 days (p=0.007). Patients who on presentation had had jaundice for >44 days (p=0.007) were unlikely to have unconjugated hyperbilirubinaemia. CONCLUSIONS: Infants with UTI may present with unconjugated hyperbilirubinaemia in the early stage. After 6 weeks, it is always conjugated hyperbilirubinaemia and is frequently associated with anaemia, elevated hepatic aminotransferases and E. coli infections.

Abstract: The incidence of urinary tract infections (UTIs) in young infants with jaundice is low, and prolonged jaundice in that setting is even rarer. In this study, we retrospectively reviewed the clinical features of 50 infants (seen from 1984 through 2004) 1 week to 3 months of age who had UTIs and prolonged jaundice and compared them with those of infants with UTIs in a similar age range at our hospital from a published study. The infants in our study had a lower incidence of high fever (12%), were less likely to have pyuria (29%), had fewer Escherichia coli but more Enterococcus species infections, and had more mixed infections (32%) than did the historical control. Klebsiella pneumoniae or Enterococcus species dominated in mixed infections (15/16) but the frequency of E. coli isolates did not differ significantly between mixed and single infections. We conclude that high fever and pyuria are unreliable criteria for screening for UTIs in young infants presenting with prolonged jaundice. Urine cultures should be obtained in such patients to determine whether a UTI is present. Mixed urinary infections, particularly K. pneumoniae and Enterococcus species, may play a key role in this entity.

Abstract: Systemic lupus erythematosus (SLE) rarely presents with massive ascites secondary to severe peritonitis before the occurrence of major diagnostic features. We described a 13-year-old girl who developed four episodes of severe abdominal pain with massive ascites. During the first episode, the patient experienced deterioration with massive ascites over a 3-day period, after a 3-month history of intermittent abdominal pain. This episode resolved under conservative treatment. During the second episode, her acute abdominal findings mimicked appendicitis or gynecologic emergency. An exploratory laparotomy was initially planned, but not performed. Thrombocytopenia postponed the surgical intervention and led to further investigation. Autoimmune studies confirmed SLE and prompted steroid therapy, which rapidly improved her painful ascites and spared an unnecessary operation. After improvement of peritonitis with ascites, and following discharge, other major clinical features of SLE ensued. The third episode of peritonitis with massive ascites responded well to steroid pulse therapy. The fourth episode of ascites was accompanied by intracranial hemorrhage that led to her death. To the best of our knowledge, this is the first case report of recurrent peritonitis with massive ascites as the initial and major manifestation of SLE in a pediatric patient.

Abstract: It is difficult to distinguish clinically between bacterial and viral causes of enterocolitis. The aim of the study was to investigate if serum cytokines can distinguish bacterial from viral enterocolitis. We prospectively enrolled 147 paediatric in-patients with acute enterocolitis. Blood was taken for leucocyte count, CRP, ESR, IL-6, IL-8, IFN-alpha and TNF-alpha on the day of admission. A pathogen was identified in 115 of the 147 children, 72 of whom had a bacterial pathogen (bacterial group) and 43 rotavirus (viral group). Mean values of the serum markers IL-6, IL-8 and CRP were significantly higher in the bacterial group. Receiver-operating characteristic curves demonstrated that a cut-off of 15 pg/ml for IL-6 had a sensitivity of 0.75 and a specificity of 0.91 for bacterial diarrhoea. Comparable values for CRP at a cut-off of 13 mg/L demonstrated a sensitivity of 0.54 and a specificity of 0.72. Values for IL-8 at a cut-off of 80 pg/ml had a sensitivity of 0.46 and a specificity of 0.71. Despite the small sample size, our data suggest that serum IL-6, IL-8 and CRP are significantly elevated in children with bacterial enterocolitis. IL-6 has a higher sensitivity, specificity and positive predictive value than IL-8 and CRP. Determination of serum cytokines might be a useful way of differentiating viral from bacterial gastro-enteritis.

Abstract: A 3-year-old boy presented at the Taiwan Adventist Hospital in Taipei with nocturnal epigastric pain and constipation. Abdominal X-ray showed colonic faecal impaction. Abdominal sonography showed gastric stasis with thickened pyloric wall and dilated rectosigmoid colons. The mouth-to-anus transit time (MATT) was prolonged. Endoscopy showed pale gastric mucosa, atony of pylorus and widening of the duodenal bulb. Three weeks after the onset of abdominal pain, he developed urinary incontinence and rapidly deteriorating paraplegia of lower limbs. Magnetic resonance imaging (MRI) showed an extradural intraspinal mass of T5-T8 and a soft tissue mass in the right superior mediastinum. After a laminectomy and tumour excision, the patient's symptoms improved quickly. The pathology revealed Burkitt's lymphoma. This is the first report of nocturnal abdominal pain and constipation as the initial manifestation of spinal Burkitt's lymphoma.

Abstract: In order to understand the childhood Salmonella enterocolitis in Taitung, we retrospectively collected the patients with the diagnosis of acute enterocolitis who were admitted to the pediatric wards of Taitung branch of Mackay Memorial Hospital from January 1, 1995 to December 31, 1998. Salmonella enterocolitis accounted for 16.8% of the total cases of acute enterocolitis. The mean age of the patients was 14.6 +/- 14.1 months old. Seventy-nine patients were male and 78 patients were female. Eighty-two patients lived in the urban area and 75 patients lived in the suburbs. The mean duration from onset of illness to admission was 2.6 +/- 1.9 days. The mean duration of hospitalization was 6.6 +/- 4.8 days. The peak incidence was in August and September. The most common clinical manifestations were fever (74.5%) and blood stool (46.5%). In 157 patients, Salmonella serogroup B was isolated from stool in 115 patients. The rate of bacteremia was 4.5%. Serogroup D accounted for 28.6% of the bacteremia. Two patients developed meningitis and one patient had osteomyelitis. Of 88 patients examined for rotavirus, 12 had positive results. There was neither bowel perforation nor mortality recorded in our study.

Abstract: Kawasaki Disease (KD) or atypical KD (AKD) rarely presents with intestinal obstruction or pseudo-obstruction. Others have reported gastrointestinal symptoms appearing with and up to 4 weeks after the occurrence of major clinical symptoms of KD. However, we presented a 1-year-old boy with prolonged fever who was found to have pyuria and liver dysfunction on the fourth day of fever. He developed a picture of intestinal pseudo-obstruction including bilious vomiting and abdominal distention on his fifth day of fever, four days before he developed the fissured lips. Because of the emergence of four major clinical criteria of KD, coronary artery dilatation, and aseptic meningitis, AKD was initially diagnosed and prompted the use of intravenous immunoglobulin on the tenth day of fever. Hydrops of the gallbladder, leukocytosis, increased ESR, and thrombocytosis were noted during hospitalization. These atypical features rarely develop all together in one patient with KD or AKD. Nevertheless, our patient had early intestinal pseudo-obstruction with almost all of the above unusual systemic manifestations and the late occurrence of clinical features of KD. We demonstrate that intestinal pseudo-obstruction in KD may develop earlier than other major clinical features and may improve under the conservative treatment. Surgery should be reserved for those who have complete intestinal obstruction presenting with significant peritoneal signs.

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Abstract: We report a case of segmental dilatation of the ileum in a 10-month-old male infant. Intermittent loose black-colored stool passage and normocytic anemia were noted at the initial visits to our hospital. There was no symptom or sign of intestinal obstruction such as abdominal distention or vomiting. On physical examination, he was found to be pale but his abdomen was soft and flat. Digital examination revealed brownish stool tinged with black-colored oil-like stool but no polyp. Laboratory studies excluded coagulopathy, hemolytic anemia and lead poisoning. During hospitalization, he was treated with nothing per mouth, intravascular fluids, ranitidine, and transfusion of packed red blood cells. All examinations including panendoscopy, Technetium-99m (99mTc)-pertechnetate Meckel's diverticulum scan, and double contrast colon series revealed no organic lesion except that 99mTc-red blood cell bleeding scans showed abnormal bleeding in the small intestine. Because of his persistent gastrointestinal bleeding with unknown cause, we did an exploratory laparotomy when the patient was 13 months old and idiopathic segmental dilatation of the ileum was confirmed. The dilated segment is supposed to be idiopathic because of histologically proven normal muscle layers without ectopic tissue. This case suggests that segmental dilatation of the ileum can only present as gastrointestinal bleeding without intestinal obstruction.

Abstract: We report a case of adrenal hemorrhage in a newborn first detected by sonography at a menstrual age of 21 weeks 4 days. An echogenic mass was noted above the right kidney. The mass became larger and hypoechoic on follow-up sonograms after birth. The mass was also seen on abdominal CT. Because it was difficult to differentiate the lesion from cystic neuroblastoma and because it shrank only modestly by 2 months after birth, we performed a surgical exploration when the patient was 2 months old, and adrenal hemorrhage was confirmed. This case suggests that adrenal hemorrhage can occur as early as the second trimester of pregnancy.