• Title/Summary/Keyword: Negative Stain

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Two cases of Listeria monocytogenes isolation from celebrospinal fluid (뇌척수액에서의 Listeria monocytogenes 분리 2예 보고)

  • Chong, Yun-Sop;Kim, Yoon-Chung;Kim, Byung-Soo;Yi, Kui-Nyung;Lee, Sam-Uel Y.
    • The Journal of the Korean Society for Microbiology
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    • v.13 no.1
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    • pp.1-5
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    • 1978
  • Listeria monocytogenes infection was considered a rather rare disease and occurs mostly either in newborn babies or in young children. However, there has been increasing reports of this infection in elderly person with various underlying disease. Recently we have experienced two cases of Listeria meningitis; one in a 4-year-old male with an acute lymphoblastic leukemia, and the other in a 43-year-old female with a breast cancer. Both were on various chemotherapeutic agents for their primary diseases when the organism, L. monocytogenes was found in their celebospinal fluid(CSF). The degree of CSF pleocytosis were quite different by cases. The former case showed a marked increase, $3,350/mm^3$, and the latter slight, $410/mm^3$, Both showed a slight decrease of CSF glucose ranging 39 to 43mg/100ml. It seems that a routine CSF analysis bears a limitted value in the diagnosis or Listeria meningitis. A direct smear of CSF with Gram's stain revealed gram-positive bacilli in one case, but none in the other. Bacterial culture of CSF yielded plenty colonies in one case, but a few in the other. It seems that isolation of L. monocytogenes must not be considered very easy, and a negative direct smear does not necessarily mean a negative culture. The two isolates we obtained showed the typical cultural and biochemical characteristics of L. monocytogenes and were found to belong to serotypes 1b and 4b. It was our experience that the identification of this organism was not very much matter because of its distinct characteristics, but the most important matter was how to think of the possibility of this organism at the begining. The two isolates were both susceptible to cephalothin, chloramphenicol, erythromycin, tetracycline and gentamicin; intermediate to ampicillin, penicillin and kanamycin; and resistant to cloxacillin.

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The Improvement of Fabrication Process for a-Si:H TFT's Yield (a-Si:H TFT의 수율 향상을 위한 공정 개선)

  • Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.11 no.6
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    • pp.1099-1103
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    • 2007
  • TFT's have been intensively researched for possible electronic and display applications. Through tremendous engineering and scientific efforts, a-Si:H TFT fabrication process was greatly improved. In this paper, the reason on defects occurring at a-Si:H TFT fabrication process is analyzed and solved, so a-Si:H TFT's yield is increased and reliability is improved. The a-Si:H TFT of this paper is inverted staggered type TFT. The gate electrode is formed by patterning with length of $8{\mu}m{\sim}16{\mu}m$ and width of $80{\sim}200{\mu}m$ after depositing with gate electrode (Cr). We have fabricated a-SiN:H, conductor, etch-stopper and photo-resistor on gate electrode in sequence, respectively. We have deposited n+a-Si:H, NPR(Negative Photo Resister) layer after forming pattern of Cr gate electrode by etch-slower pattern. The NPR layer by inverting pattern of upper Sate electrode is patterned and the n+a-Si:H layer is etched by the NPR pattern. The NPR layer is removed. After Cr layer is deposited and patterned, the source-drain electrode is formed. The a-Si:H TFT made like this has problems at photo-lithography process caused by remains of PR. When sample is cleaned, this remains of PR makes thin chemical film on surface and damages device. Therefor, in order to improve this problem we added ashing process and cleaning process was enforced strictly. We can estimate that this method stabilizes fabrication process and makes to increase a-Si:H TFT's yield.

Study of Serious Bacterial Infections in Febrile Infants Younger than Three Months of Age (열이 있는 3개월 이하의 영아에서 세균성 감염의 예측에 대한 연구)

  • Jeon, Myeoung Won;Lee, Ji Young;Jang, Young Taek
    • Pediatric Infection and Vaccine
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    • v.10 no.2
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    • pp.215-222
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    • 2003
  • Purpose : This study was to analyze serious bacterial infections in infants younger than three months of age and to review the direction of treatments for these patients. Methods : 378 febrile infants with a rectal temperature ${\geq}38.0^{\circ}C$ visited from Jan. 2001 through Dec. 2002 were retrospectively studied. Infants with the following criteria belonged to the low risk group. WBC $5,000{\sim}15,000/mm^3$, WBC negative in urine stick test and negative for nitirite test, CSF WBC < $10/mm^3$ and negative in CSF gram stain, negative chest X-ray, stool WBC <5/HFP(high power field), and focal infection. If any of the above criteria were not met, they belonged to the high risk group. SBI was defined as a positive culture of urine, blood or CSF. SI was defined as aseptic meningitis or pneumonia including above laboratory tests of SBI. SBI patients were separately compared with two groups, high risk and low risk. Results : Of the 378 infants that were tested 216(57.1%) were in the high risk group and 162(42.9%) in the low risk group. Among 105 SBI(27.8%) and 172 SI(45.5%), there were 98 urinary tract infection(25.2%), 10 bacteremia(2.6%), 2 bacterial meningitis(0.6%), and 77 aseptic meningitis(22.8%). There were 76 SBI(35.2%) from the high risk group and 29 SBI(17.9%) from the low risk group identified. The results of the sensitivity(72.4%), the specificity(48.7%), the negative predictive value(82.1%) and the positive predictive value (35.2%) were calculated. Conclusion : Even though the probability of SBI in the low risk group is insignificant, it should still be considered in febrile infants younger than 3 months of age. I believe the CSF study is necessary because of the moderate high incidence of abnormal finding in our study.

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The Clinical Significance of Cathepsin D and p53 Expression in Locally Advanced Rectal Cancer (국소진행된 직장암에서 Cathepsin D와 p53 발현의 임상적 의의)

  • Kim, Jun-Sang;Lee, Sheng-Jin;Kim, Jin-Man;Cho, Moon-June
    • Radiation Oncology Journal
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    • v.26 no.1
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    • pp.56-64
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    • 2008
  • Purpose: Cathepsin D(CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer(cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57(64%) of the tumors and 32(35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression(p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival(OS) and disease-free survival(DFS) rates were not different between tumor CD-negative and positive patient biopsy samples(69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients(p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.

The Study of Mycoplasma pneumoniae Infection in Children with Respiratory Tract Infection (호흡기 감염증 소아에서 Mycoplasma pneumoniae 감염에 관한 연구)

  • Cho, Jung Ik;Kim, Jong Geon;Kim, Jong Ho;Cho, Ji Hyun;Kim, Jong Duck
    • Pediatric Infection and Vaccine
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    • v.9 no.1
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    • pp.85-94
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    • 2002
  • Purpose: This study was performed for analysis of the results of polymerase chain reaction(PCR) and antibody test of Mycoplasma pneumoniae(M. pneumoniae) in children with symptoms of respiratory tract infection. In the cases of both positive antibody test and PCR for M. pneumoniae, the chest X-ray findings were assessed. Methods: The antibody test was done in 1,979 cases who have been admitted to Wonkwang university hospital department of pediatrics with symptoms of respiratory tract infection from January, 2000 to December, 2001. The positive antibody test was defined as titer of 1 : 80 and over 1 : 80. The PCR of M. pneumoniae were done in randomly selected 131 cases of respiratory tract infection. The chest X-ray findings were assessed in the cases of positive antibody test and PCR. Results: The numbers of cases of the positive antibody test for M. pneumoniae were 499 cases(25%). The PCR for M. pneumoniae were performed in 131 cases and the 45 cases(34%) were positive and 86 cases(66%) were negative. The 56 of 86 PCR negative cases were also negative antibody test, but 30 cases were positive antibody test. The 36 cases of 45 PCR positive cases were antibody positive, and 9 cases were antibody negative. The sputum Gram stain and culture for M. pneumoniae were negative in all the 499 cases of mycoplasma antibody positive respiratory infection. In these antibody positive 499 cases, the most common X-ray findings was interstitial pneumonic infiltration in 266 cases(53%), and pleural effusion were detected in 22 cases(4%), but nonspecific chest X-ray finding showed in 129 cases(26%). In PCR positive 45 cases, the most common chest X-ray finding was interstitial pneumonic infiltration in 32 cases(71%). Conclusion: The PCR for M. pneumoniae is more useful method for detection of mycoplasma infection in children with respiratory tract infection. The M. pneumoniae is a important etiologic agent for respiratory infection in children.

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The Clinical Features of Endobronchial Tuberculosis - A Retrospective Study on 201 Patients for 6 years (기관지결핵의 임상상-201예에 대한 후향적 고찰)

  • Lee, Jae Young;Kim, Chung Mi;Moon, Doo Seop;Lee, Chang Wha;Lee, Kyung Sang;Yang, Suck Chul;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo;Lee, Jung Hee
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.5
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    • pp.671-682
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    • 1996
  • Background : Endobronchial tuberculosis is definded as tuberculous infection of the tracheobronchial tree with microbiological and histopathological evidence. Endobronchial tuberculosis has clinical significance due to its sequela of cicatrical stenosis which causes atelectasis, dyspnea and secondary pneumonia and may mimic bronchial asthma and pulmanary malignancy. Method : The authors carried out, retrospectively, a clinical study on 201 patients confirmed with endobronchial tuberculosis who visited the Department of Pulmonary Medicine at Hangyang University Hospital from January 1990 10 April 1996. The following results were obtained. Results: 1) Total 201 parients(l9.5%) were confirmed as endobronchial tuberculosis among 1031 patients who had been undergone flexible bronchofiberscopic examination. The number of male patients were 55 and that of female patients were 146. and the male to female ratio was 1 : 2.7. 2) The age distribution were as follows: there were 61(30.3%) cases in the third decade, 40 cases(19.9%) in the fourth decade, 27 cases(13.4%) in the sixth decade, 21 cases(10.4%) in the fifth decade, 19 cases(9.5%) in the age group between 15 and 19 years, 19 cases(9.5%) in the seventh decade, and 14 cases(7.0%) over 70 years, in decreasing order. 3) The most common symptom, in 192 cases, was cough 74.5%, followed by sputum 55.2%, dyspnea 28.6%, chest discomfort 19.8%, fever 17.2%, hemoptysis 11.5%, in decreasing order, and localized wheezing was heard in 15.6%. 4) In chest X-ray of 189 cases, consolidation was the most frequent finding in 67.7%, followed by collapse 43.9%. cavitary lesion 11.6%, pleural effusion 7.4%, in decreasing order, and there was no abnormal findings in 3.2%. 5) In the 76 pulmanary function tests, a normal pattern was found in 44.7%, restrictive pattern in 39.5 %, obstructive pattern in 11.8%, and combined pattern in 3.9%. 6) Among total 201 patients, bronchoscopy showed caseous pseudomembrane in 70 cases(34.8%), mucosal erythema and edema in 54 cases(26.9%), hyperplastic lesion in 52 cases(25.9%), fibrous s.enosis in 22 cases(10.9%), and erosion or ulcer in 3 cases(1.5%). 7) In total 201 cases, bronchial washing AFB stain was positive in 103 cases(51.2%), bronchial washing culture for tuberculous bacilli in 55 cases(27.4%). In the 99 bronchoscopic biopsies, AFB slain positive in 36.4%. granuloma without AFB stain positive in 13.1%, chronic inflammation only in 36.4%. and non diagnostic biopsy finding in 14.1%. Conclusions : Young female patients, whose cough resistant to genenal antitussive agents, should be evaluated for endobronchial tuberculosis, even with clear chest roentgenogram and negative sputum AFB stain. Furthermore, we would like to emphasize that the bronchoscopic approach is a substantially useful means of making a differential diagnosis of atelectasis in older patients of cancer age. At this time we have to make a standard endoscopic classification of endobronchial tuberculosis, and well designed prospective studies are required to elucidate the effect of combination therapy using antituberculous chemotherapy with steroids on bronchial stenosis in patients with endobronchial tuberculosis.

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Helicobacter pylori reinfection rate by a 13C-urea breath test and endoscopic biopsy tests in Korean children (한국 소아에서 Helicobacter pylori 박멸 후 13C-요소 호기 검사와 내시경적 생검을 이용한 재감염률 연구)

  • Shim, Jeong Ok;Seo, Jeong Kee
    • Clinical and Experimental Pediatrics
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    • v.49 no.3
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    • pp.268-272
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    • 2006
  • Purpose : The reinfection rate of H. pylori reported before $^{13}C$-urea breath test($^{13}C$-UBT) era was higher than that of the post $^{13}C$-UBT era. Children are usually reluctant to receive invasive endoscopic evaluation for the reinfection of H. pylori, particularly when they are asymptomatic. The aim of the study is to discover the reinfection rate by different diagnostic tests, and to find out what causes the difference. Methods : Children confirmed to be eradicated from H. pylori were included in the study. Reinfection was evaluated by endoscopic biopsy based tests(n=34, mean age $11.5{\pm}3.7$ years) and/or a $^{13}C$-UBT(n=38, mean age $10.0{\pm}3.6$ years) at the time of 18 months after eradication. At first visit, H. pylori infection had been diagnosed by positive results from a rapid urease test, Giemsa stain and Warthin-Starry stain and/or a positive culture. Eradication was defined as negative results from all above tests 1-3 months after eradication therapy. Results : Reinfection rate by endoscopic biopsy based tests was 35.3 percent(12/34). All patients had abdominal symptoms(P=0.000). Reinfection rate was 13.2 percent(5/38) by a $^{13}C$-UBT. Reinfection rate was higher in children with abdominal symptoms(P=0.008). There was no evidence that reinfection rate depended on the sex(P=0.694), age(P=0.827), diseases(peptic ulcers vs gastritis, P=0.730) and eradication regimen(P=0.087). Conclusion : Helocibacter pylori reinfection rate in Korean children was 13.2 percent per 18 months by a non-invasive test or $^{13}C$-UBT. Accurate determinations of the reinfection rate in children is affected by the compliance of the diagnostic tests. Non-invasive tests should be considered to investigate the reinfection rate in children.

In Vitro Effect of 808-nm Diode Laser on Proliferation and Glycosaminoglycan Synthesis of Rabbit Articular Chondrocytes (토끼 관절 연골세포의 증식과 글리코스아미노글리칸 합성에 대한 808-nm 다이오드 레이저의 효능 평가)

  • Minar, Maruf;Hwang, Ya-won;Choi, Seok-hwa;Kim, Gonhyung
    • Journal of Veterinary Clinics
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    • v.32 no.4
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    • pp.295-300
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    • 2015
  • The aim of the study was to assess the in vitro effect of 808-nm InGaAs diode laser on rabbit articular chondrocyte proliferation and sulphated glycosaminoglycan (sGAG) synthesis in alginate bead. Previous studies revealed either positive or negative stimulatory effects of laser on different types of cells. A 808-nm InGaAs diode laser at 1.0W power output was used to irradiate the rabbit chondrocytes in alginate beads with energy densities of $31J/cm^2$ (G 1) and $62J/cm^2$ (G 2) corresponding to the experimental groups for 10 seconds and 20 seconds, respectively at 24, 48, 72 and 96 hours after seeding. Control group was left untreated. MTT assay was performed at 1 week and 2 weeks after the $1^{st}$ laser irradiation in alginate beads. sGAG synthesis in alginate beads at 1 week and 2 weeks were determined by DMMB assay. Histological evaluation for cellular distribution and sGAG deposition around the cells were performed by alcian blue stain. MTT assay revealed no positive stimulatory effect in cell proliferation in alginate bead. DMMB assay results showed significantly increased sGAG production in G 2 chondrocytes at 2 weeks. Image analysis of alcian blue stained slides also showed significantly higher percentage of positive alcian blue stain in G 2 chondrocytes. This result suggests that 808-nm InGaAs diode laser with 1.0 W power output although cannot stimulate cell proliferation it can increase the cell secretion activity and sGAG deposition in alginate beads.

The Clinical Utility of Polymerase Chain Reaction in the Bronchoalveolar Lavage Fluid for the Detection of Mycobacteria (객담 도말 음성인 환자에서 기관지폐포 세척액 결핵균 중합 효소 연쇄반응 검사의 유용성)

  • Mo, Eun-Kyung;Kyung, Tae-Young;Kim, Dong-Gyu;Park, Myung-Jae;Lee, Myung-Goo;Hyun, In-Gyu;Jung, Ki-Suck;Lee, Kyung-Wha
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.3
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    • pp.519-528
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    • 1998
  • Background: Diagnosis of pulmonary tuberculosis is not easy when the sputum smear for Mycobacterium tuberculosis(M. Tb) is negative. We evaluated the clinical utility of polymerase chain reaction(PCR) for detecting M. Tb in bronchoalveolar lavage(BAL) samples. Methods: We recruited 84 patients whose sputum smear for M. Tb were negative or not available due to no production of sputum. We performed bronchoalveolar lavage for acid-fast stain, culture of mycobacteria, and PCR assay of BAL fluid. We analyzed the results of microbiologic examination. Results: The sensitivity of BAL fluid smear, culture, and PCR were 20%, 38%, and 40%, respectively. The specificity of BAL fluid PCR was 95%. The positive predictive value of PCR was 89%. The smear of BAL fluid was positive in 17%. The PCR of BAL fluid was the only diagnostic test in 17%. Therefore, the BAL fluid analysis including smear and PCR was diagnostic in 34 % within 24 hours. The BAL fluid analysis including smear, PCR, and culture was diagnostic in 55% within 2 month. Conclusion: The BAL fluid PCR was valuable method in the diagnosis of pulmonary tuberculosis in patients whose sputa were not available or reveal negative smear.

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Study on Persistent Infection of Japanese Encephalitis Virus Beijing-l Strain in Serum-free Sf9 Cell Cultures

  • Kim, Hun;Lee, Su-Jeen;Park, Jin-Yong;Park, Yong-Wook;Kim, Hyun-Sung;Kang, Heui-Yun;Hur, Byung-Ki;Ryu, Yeon-Woo;Han, Sang-In
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.25-31
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    • 2004
  • Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.