Biomedical subjects

Tao Hong

Publications and source records attributed to Tao Hong.

At least 37 records · Page 2Linked to original sources

PubMed2005

Effects of specific interleukin-1beta-converting enzyme inhibitor on ischemic acute renal failure in murine models.

AIM: To study the effect of selective interleukin-1beta-converting enzyme (ICE, caspase-1) inhibitor on ischemic acute renal failure (ARF). METHODS: Mouse models of ischemic ARF were treated with the specific ICE inhibitor AC-YVAD-CMK. A renal function assay and renal morphological studies were employed to estimate the renal protective effect of AC-YVAD-CMK. The survival rate of mouse models was also analyzed by a time series test. Furthermore, renal ICE activity, mature interleukin-18 (IL-18) protein expression and interferon-gamma (IFN-gamma) mRNA expression were also detected by fluorescent enzyme-linked immunosorbent assay (ELISA), ELISA, and semi-quantitative reverse transcription-polymerase chain reaction, respectively. RESULTS: The levels of blood urea nitrogen (BUN) and serum creatinine (Scr) increased remarkably in the model controls compared with the sham-operated groups (P<0.01). Typical renal tubular necrosis was found in the model controls. Renal ICE activity, mature IL-18 protein expression, and IFN-gamma mRNA expression were also increased significantly in the model controls compared with the sham-operated groups. The levels of BUN and Scr in the AC-YVAD-CMK therapy group were decreased significantly compared with the untreated model controls (P<0.01). Renal tubulointerstitial lesion was also attenuated significantly (P<0.05). AC-YVAD-CMK therapy alleviated the clinical features of ARF, and increased the survival rate (P<0.01). Furthermore, AC-YVAD-CMK therapy also decreased ICE activity, mature IL-18 protein expression, and IFN-gamma mRNA expression in renal tissue (P<0.05). CONCLUSION: The selective ICE inhibitor AC-YVAD-CMK can effectively protect the kidney from acute ischemic lesions. This protective effect is associated with decreased renal ICE activity and suppressed IL-18 maturation and IFN-gamma mRNA transcription.

Acute Kidney Injury↗

PubMed2005

[Study on the RNA of severe acute respiratory syndrome (SARS) associated coronavirus in the blood and excretion of convalescent patients with SARS].

OBJECTIVE: To examine the RNA of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) in the blood and excretion of convalescent patient with SARS for prevention and treatment of the disease. METHODS: A total of 276 samples, including plasma, urine, feces and sputum, obtained from 23 convalescent patients with SARS were studied at 3 time-points at least 21 days after the onset of symptoms. RNA was extracted and nested reverse transcription-polymerase chain reaction (RT-PCR) was carried out using SARS-CoV specific primers. RESULTS: Among the 276 samples, SARS-CoV RNA was detected in 6 cases (38.8%) by nested RT-PCR. The positive rates of SARS-CoV RNA was 5.8% in feces and 2.9% in sputum samples but SARS-CoV RNA was not detectable in plasma and urine of all the cases. CONCLUSION: The existence of SARS-CoV RNA in the excretion of some convalescent patients with SARS showed that the excretion from these patients should be carefully treated whilthe re-transmission of SARS by which, should be further studied.

Adolescent↗

PubMed2005

[Receptor-binding ability of fragments 260-600 and 397-796 of SARS-associated coronavirus spike protein].

BACKGROUND: To investigate the interaction between the host cell and the truncated S fragments to identify the receptor-binding domain of the spike (S) protein of SARS-associated coronavirus (SARS-CoV). METHODS: Two different fragments S260-600 and S397-796 of the SARS-CoV S protein were expressed in Escherichia coli (E.coli) using a pET expression vector, respectively. The two recombinant proteins were separately verified by Western blot, purified by nickel-affinity chromatography, and incubated with Vero cells, a susceptible cell line of SARS-CoV infection, for cell binding assay. After the sequential probing with sera from convalescent SARS-patients and FITC-labeled anti-human IgG, the cells were analyzed by flow cytometry. The NIH 3T3 cell, a non-permissive cell line of SARS-CoV infection, was used as controls. RESULTS: The recombinant proteins S260-600 and S397-796 were efficiently expressed in an insoluble form in E.coli. The appropriate expression of the proteins was confirmed by Western blotting using both SARS patients' sera and anti-6 x histidine antibody. The flow cytometry results showed that the both proteins were able to bind Vero cells, but the binding ability of S260-600 was somewhat stronger than that of S397-796. In contrast, the S260-600 protein did not bind NIH3T3 cells. CONCLUSION: Both S260-600 and S397-796 exhibited different receptor binding activity. The S260-600 fragment probably contains the important receptor binding domain and could be a potential candidate for the development of SARS vaccine and anti-SARS therapeutics.

Animals↗

PubMed2004

[Preliminary study of immunity and safety of recombinant adenovirus expressing rotavirus structural proteins in rhesus monkeys].

OBJECTIVE: To preliminarily evaluate the immunity and safety of the recombinant adenoviruses expressing rotavirus structural proteins VP7 and VP6 in rhesus monkeys to lay a foundation for the development of novel genetic engineering vaccine against rotavirus. METHODS: Baby monkeys were immunized with the recombinant adenoviruses intranasally or orally. Serum IgG against rotavirus was measured with ELISA. During the course of the immunization, besides the daily monitoring of body temperature, weight and clinical symptoms, the routine blood and urine tests and liver and kidney function tests were also conducted. RESULTS: Monkeys immunized via intranasal or oral routes could both generate serum IgG against rotavirus. During the immunization, the temperature of monkeys was normal and body weight raise stably. Both routine blood and urine tests and liver and kidney function tests showed no significant alteration compared with the control group. CONCLUSION: The immunization with the recombinant adenoviruses expressing rotavirus antigens is able to induce rotavirus specific efficient immune responses and is safe to baby rhesus monkeys. The preliminary results implied that the recombinant adenoviruses could be an ideal vaccine for rotavirus and lay a foundation for further studies.

Adenoviridae↗

PubMed2004

[Orthotopic heart transplantation: clinical experience on 56 patients].

OBJECTIVE: To summarize the clinical experience on 56 patients undergoing orthotopic heart transplantation. METHODS: Between May 2000 and December 2003 56 patients, 47 with cardiomyopathy, 2 with end-stage valvular heart disease, 2 with end-stage ischemic heart disease, 2 with primary malignant cardiac tumor, 1 with complicated congenital heart disease, 1 with muscular dystrophy cardiomyopathy, and 1 with refractory malignant ventricular arrhythmias, underwent orthotopic heart transplantation in the Transplantation Center of Fudan University. The operative procedures included 19 conventional Stanford orthotopic cardiac transplantation in 19 cases and bicaval anastomotic cardiac transplantation in 37 cases. Postoperatively, the patients were prescribed with cyclosporine A + corticosteroids + MMF or FK506 + corticosteroids + MMF as anti-rejection therapy. RESULTS: One patient undergoing his fifth operation died of bleeding 3 days after operation. All survivors were followed-up for 12.4 months on average. Five patients died within 1 year postoperatively with a one-year survival rate of 91%. One patient died of allograft vasculopathy 38 months after operation. Other patients enjoyed heart function recovering to class I-II (NYHA). CONCLUSION: With proper recipient selection, excellent donor heart conservation, bicaval anastomotic technique, and efficient postoperative surveillance and treatment, heart transplantion may produce satisfying mid-term results.

Adolescent↗

PubMed2004

[Expression of chemokine-like factor 2(CKLF2) mRNA in rat myocardium with hypertrophy].

OBJECTIVE: To investigate the expression of CKLF2 mRNA in the rat myocardium with hypertrophy. METHODS: Rat abdominal aorta was constricted by operation to make myocardium hypertrophy in the hypertrophy group and the control group was subjected to the sham-operation. The CKLF2 mRNA expressions were tested by competitive polymerase chain reaction (CPCR). RESULTS: Compared to the control-group, the expression of CKLF2 mRNA of hypertrophy-group significantly increased on the 17th day after operation, and decreased to the control level on the 45th day. CONCLUSION: The expression of rCKLF2 mRNA in rat myocardium with hypertrophy significantly increased in early days of hypertrophy.

Animals↗

PubMed2004

[Construction and expression of the fusion protein DT389-hIL-13 and its cytotoxicity to glioma cell lines].

OBJECTIVE: To construct a fusion protein toxin DT389-hIL-13 which comprises the N-terminal 389 amino acids of diphtheria toxin (DT389) and human interlukin 13 (hIL-13), and to explore its cytotoxicity on U251 glioma cells. METHODS: The cDNA of hIL-13 gene was amplified by PCR and linked with the 3'-terminus of the gene encoding the N-terminal 389 amino acids, which correspond to the enzyme domain and transmembrane domain of diphtheria toxin. The tandem constructed gene was then inserted into an E. coli expression vector pET30a. The resulted expression vector was transformed into E. coli BL21 and induced by IPTG. The expressed protein was analyzed by SDS-PAGE and Western blot analysis. U(251) glioma cells were cultured DT389-hIl-13 was added into the culture. The cytotoxicity was determined using colorimetric MTS proliferation assay. RESULTS: The expression plasmid pET30a/DT389-hIL13 was constructed with correct sequence. The recombinant protein was successfully expressed in E. coli in manner of inclusion body and with a relative molecular weight of about 55 000, which reacted well with both anti-diphtheria toxin and anti-hIL-13 polyclonal antibody in Western blot assay. The purified recombinant chimeric toxin was found to effectively inhibit the prolifieration of glioblastoma multiforme cells bearing high affinity hIL-13 receptors, and resulted in dose-dependent relationship with 50% inhibition concentration (IC(50)) of 5 x 10(-)11mol/L. CONCLUSION: Prokaryotic expression system can be recruited to produce recombinant chimeric toxin DT389-hIL-13. The results may lay a foundation for preparing specific the agent targets for tumors overexpressing IL-13 receptor.

Brain Neoplasms↗

PubMed2004

Isoflavones stimulate estrogen receptor-mediated core histone acetylation.

The isoflavones genistein and daidzein and the daidzein metabolite equol have been reported to interact with estrogen receptors (ERs). Some studies indicate that they behave clinically like estrogen in some estrogen-deficiency diseases. However, the detailed molecular mechanism used by these compounds to create beneficial effects in patients with estrogen-related diseases has not been clarified. Using histone acetyltransferase (HAT) assay, we found that equol, genistein, and AglyMax had significant effects on ERalpha-mediated histone acetylation. Although 17beta-estradiol (E2)-dependent HAT activity of steroid receptor coactivators 2 (SRC2) and p300 mediated by ERbeta could be detected, it was weaker than that mediated by ERalpha. Equol, genistein, AglyMax, and daidzein all markedly stimulated ERbeta-mediated histone acetylation. On the other hand, anti-estrogenic compounds ICI 182,780 (ICI) and tamoxifen (TA) did not have an effect on HAT activity mediated by either ERalpha or ERbeta. Our data indicate that estrogenic ligands exert their effects by elevating histone acetylation and coactivator activity of ER, and suggest that the risk of estrogen-related diseases might be reduced by a sufficient amount of genistein or AglyMax supplements.

Acetylation↗

PubMed2004

[The protective effect of interleukin-1 receptor antagonist on postischemic reperfused myocardium and its possible mechanism].

OBJECTIVE: To observe the dynamic changes of plasma inflammatory cytokine interleukin-1 beta (IL-1 beta) in patients with acute myocardiac infarction (AMI) before and after recanalization of the infarct related artery (IRA) and to observe the effect of recombinant human IL-1 receptor antagonist (rhIL-1ra) on the postischemic reperfused myocardium in experimental rabbits. METHODS: (1) ELISA was used to measure the plasma IL-1 beta of 22 AMI patients, 20 males and 2 females, aged 64 +/- 12, before emergency percutaneous coronary intervention (PCI), and 12 hours and 24 hours after-intervention, and measure the plasma IL-1 beta of 8 healthy controls, 6 males and 2 females, aged 56 +/- 9. (2) Forty rabbits underwent ligation of the left circumflex branch of coronary artery (LCX) for 50 minutes and reperfusion for 4 hours after the ligatures were untied. The rabbits were randomly divided into 4 groups of 10 rabbits to be injected into the left ventricle immediately before the reperfusion with rhIL-1ra 10 mg/kg (group A), 20 mg/kg (group B), or 40 mg/kg (group C), and normal saline (control group) respectively. After reperfusion of 4 hours, the LCX was re-ligated. Evans blue was injected into the left ventricle. 15% KCl was injected intravenously to kill the rabbits. Their hearts were taken out to weigh the non-ischemic, ischemic, and necrotic cardiac muscles so as to calculate the infarct size. The myoperoxidase (MPO) activity was measured by colorimetry. Sections of myocardium were made. The number of apoptotic cardiomyocytes was evaluated by TUNEL method. The apoptotic rate of cardiomyocyte was measured by annexin V method. The DNA expression of myocardium was detected by DNA laddering method. The expressions of Bcl-2 and Bax apoptosis genes were assessed. RESULTS: (1) The average plasma IL-1 beta level of the 22 AMI patients before emergency PCI was significantly higher than that of the controls (28 pg/ml +/- 9 pg/ml vs. 20 pg/ml +/- 11 pg/ml, P < 0.05), and became the highest 12 hours after the intervention (86 pg/ml +/- 14 pg/ml), and the high level lasted to 24 hours after emergency PCI. (2) In the ischemia-reperfusion rabbit model, the infarct size was 47% +/- 7% in the group A, 34% +/- 8% in the group B, 31% +/- 6% in the group C, and 61% +/- 11% in the control group (P < 0.05, 0.01, and 0.01 respectively). The activity of myocardial MPO was 16.6 +/- 3.6 min(-1).g.w.w(-1) in the group A, 10.9 min(-1).g.w.w(-1) +/- 1.9 min(-1).g.w.w(-1) in the group B, 7.8 min(-1).g.w.w(-1) +/- 2.2 min(-1).g.w.w(-1) in the group C, and 20.5 min(-1).g.w.w(-1) +/- 4.5 min(-1).g.w.w(-1) in the control group (P < 0.05, 0.01, and 0.01 respectively). The cardiomyocyte apoptosis evaluated by TUNEL was 38.3 n/HP +/- 7.4 n/HP in the group A, 25.6 n/HP +/- 6.8 n/HP in the group B, 12.2 n/HP +/- 3.3 n/HP in the group C, and 44.4 n/HP +/- 9.5 n/HP in the control group (P < 0.05, and P < 0.01 respectively in comparison between the group B and the control group and between the group C and the control group). The apoptotic rate by annexin V method was 11.6% +/- 2.7% in the group A; 7.7% +/- 2.4% in the group B, 4.7% +/- 1.4% in the group C, and 15.6% +/- 3.5% in the control group (P < 0.05, 0.01, and 0.01 respectively). DNA electrophoresis showed scaling ladder pattern only in the control group. The fluorescent density of the apoptosis gene Bax in myocardium was 24.9 +/- 8.2 in the group A; 15.5 +/- 3.4 in the group B, 10.6 +/- 2.5 in the group C, and 33.3 +/- 9.4 in the control group (P = 0.0298, 0091, and 0052 respectively) and no significant difference in the expression of Bcl-2 was shown among the 4 groups. Myocardial MPO was correlated with cardiomyocyte apoptosis (r = 0.86 by TUNEL, P < 0.01; r = 0.75 by Annexin V method, P < 0.05). CONCLUSION: Inflammatory cytokine IL-1 beta is involved in myocardial ischemia-reperfusion injury. With potential therapeutic value in prevention and treatment of ischemia-reperfusion injury to myocardium, rhIL-1ra may reduce myocardial ischemia-reperfusion injury by suppression of cardiomyocytes apoptosis mediated by IL-als; 0.86 by TUNEL, P < 0.01; r = 0.75 by Annexin V method, P < 0.05). CONCLUSION: Inflammatory cytokine IL-1 beta is involved in myocardial ischemia-reperfusion injury. With potential therapeutic value in prevention and treatment of ischemia-reperfusion injury to myocardium, rhIL-1ra may reduce myocardial ischemia-reperfusion injury by suppression of cardiomyocytes apoptosis mediated by IL-1.

Acute Disease↗

PubMed2004

[Infection after cardiac transplantation: prevention and management].

OBJECTIVE: To explore and sum up the characteristics of infection after cardiac transplantation and to discuss the prophylaxis and management. METHOD: From May 2000 to April 2003, 36 patients received orthotopic heart transplantation, the clinical data were observed and analyzed. RESULTS: Infection occurred in 2 (6%) cases, both belonged to lung infection caused by human cytomegalovirus. The 2 cases were cured by ganciclovir intravenously. CONCLUSION: Good prophylactic method may decrease post cardiac transplantation infection significantly. It is very important to early diagnose and treat infection.

Adolescent↗

PubMed2004

[Relation between inflammation related cytokines and myocardial microcirculatory reperfusion state].

OBJECTIVE: To observe the dynamic fluctuation of inflammatory cytokines (IL-1beta, TNFalpha) and anti-inflammatory cytokine (IL-10) in acute myocardial infarct (AMI) patients before and after recanalization of infarct related artery (IRA) and analyze the relationship between fluctuation of cytokines and reperfusion state of myocardial tissue. METHODS: (1) In 22 AMI patients and 8 healthy subjects, plasma IL-1beta, TNFalpha and IL-10 were measured with ELISA before emergency percutaneous coronary interposition (PCI), 12 h and 24 h post-intervention. (2) The 22 AMI patients were further divided into 2 groups according to ST-segment change in ECG at 2h after reperfusion: group A, good reperfusion of myocardium, ST decrease >or= 70% (n = 12) and group B, poor reperfusion of myocardium, ST decrease < 70% (n = 10). The change of plasma levels of IL-1beta, TNFalpha and IL-10 of the two groups was compared. RESULTS: (1) Plasma TNFalpha and IL-10 in group A and B were not significantly higher those that the healthy controls (P > 0.05) before emergency PCI, but IL-1beta was significantly higher (P < 0.05). Plasma IL-1beta and TNFalpha in group A and B and IL-10 in group B at 12 h and 24 h post-intervention were significantly higher than those before PCI (P < 0.01, P < 0.05, P < 0.05), while IL-10 in group A was not (P > 0.05). (2) Plasma IL-1beta, TNFalpha and IL-10 in group B were not higher than those in group A before PCI, but they were significantly higher at 12 h post-PCI (P < 0.01, P < 0.05, P < 0.05) and IL-1beta and IL-10 were still higher at 24 h post-PCI (P < 0.05). The increment of IL-10 was significantly less than that of IL-1beta and TNFalpha in group A and B (P < 0.01, P < 0.05). CONCLUSIONS: Cytokines may be involved in myocardial ischemia-reperfusion injury and increase of inflammatory cytokines may be a marker of poor myocardial microcirculatory reperfusion.

Adult↗

PubMed2004

Dynamic analyses of PrP and PrP(Sc) in brain tissues of golden hamsters infected with scrapie strain 263K revealed various PrP forms.

OBJECTIVE: To expatiate dynamic changes in hamsters infected with scrapie strain 263K, to observe the presence and aggravation of various forms of PrP and PrP(Sc) during incubation period, and to probe primarily the relationship between the onset of clinic manifestations and the presence of different PrP(Sc) forms. METHODS: Hamster-adapted scrapie strain 263K was intracerebrally inoculated into hamsters. Different forms of PrP and PrP(Sc) were monitored dynamically by Western blot and immuno-histochemical assays. The presence of scrapie-associated fibril (SAF) was assayed by electron microscopy analysis (EM) and immuno-golden EM. RESULTS: PrP(Sc) was initially detected in the brain tissues of the animals in 20 days post-inoculation by immunohistochemistry and 40 days with Western blot. Quantitative evaluations revealed that the amounts of PrP and PrP(Sc) in brain tissues increased along with the incubation. Several high and low molecular masses of PrP were seen in the brains of the long-life span infected animals. Deglycosylation assays identified that the truncated PrP in the infected brains showed similar glycosylation patterns as the full-length PrP. The presence of short fragments was seemed to relate with the onset of clinical conditions. CONCLUSION: These results indicate that infectious agents exist and accumulate in central nerve system prior to the onset of the illness. Various molecular patterns of PrP(Sc) may indwell in brain tissues during the infection.

Animals↗

PubMed2004

[Expression of main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses].

OBJECTIVE: To express main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses on the basis of previous investigation of the prevalence of rotavirus in China. METHODS: On the basis of successfully expression of human rotavirus protein G1VP7 by recombinant adenovirus vector, the authors constructed some potent recombinant adenovirus strains encoding rotavirus G2 VP7 and G3 VP7 genes which belong to the main rotavirus isolates 97S43 and 97S48. RESULTS: Replication defective recombinant adenoviruses expressing human rotavirus serotype G2 and G3 VP7 genes, named as rvAdG2VP7 and rvAdG3VP7 were successfully constructed. VP7 genes integrated into the viral genome were identified by PCR and Southern blot assay, and specific transcription were detected by RT-PCR in the 293 cells infected with recombinant adenoviruses. Expression of rotavirus VP7 proteins was demonstrated by Western blot assay. CONCLUSION: The established recombinant adenoviruses expressing G2 and G3 serotype VP7s laid a significant basis for further animal experiments in the development of multivalent rotavirus vaccines against rotavirus infection.

Adenoviridae↗

PubMed2004

[Preparation of heart valve scaffold and cell seeding].

To prepare scaffolds for heart valve tissue engineering, porcine heart valves were treated with varied concentrations of trypsin for 32, 56, 80 and 104 h or followed with DNase. And then the structure of acellular valves was observed under light microscope, scanning and transmission electron microscope. Porcine endothelial cells, human endothelial cells, and canine myofibroblasts were reseeded onto the acellularized porcine heart valve scaffolds once a day for 3 days. The valves were analyzed by immunohistochemical staining and electron microscopy. Results show that all endothelial cells and the majority of interstitial cells were removed from the heart valves after digestion with trypsin for 104 h, and the collagen fiber structure remains intact, but the space between collagen fibers increased slightly. Incubation with trypsin for 80 h and then with DNase almost removed all cells, and the collagen fiber structure and the space between the fibers remain intact. After reseeding, human endothelial cells almost fully cover the valve scaffold surface as shown by H-E staining and platelet endothelial cell adhesion molecules (PECAM-1) staining. Xenogeneic porcine endothelial cells also adhered to and grew on the scaffolds. As shown by H-E staining and actin staining, canine myofibroblasts not only adhered to the surface of valve scaffold but also migrated to the inner part of matrix after one week culture. These results suggest that the digestion of porcine heart valves with trypsin combining with DNase is a suitable method to remove cells. The acellular porcine heart valve scaffolds have a quite favorable biocompatibility with human and porcine endothelial cells as well as canine myofibroblasts.

Animals↗

PubMed2004

[Expression, purification and specific monoclonal antibodies preparation of diphtheria toxin A fragment].

Diphtheria toxin A fragment (DTA) is an essential catalytic domain of diphtheria toxin (DT)-based immunotoxin. DTA protein and its antibodies play an important role in the studies on toxicology, purification and identification of DT-based immunotoxins. In this paper, DTA was expressed and purified from E. coli. After Q-Sepharose FF chromatography and (Ni+)-Sepharose affinity chromatography, 6 x His-DTA fusion protein with 90% purity was achieved. Using the purified DTA as antigen to immunize BalB/c mice, 2 hybridoma cell lines (designated as 3B6 and 3B9, respectively) secreting monoclonal antibodies (McAbs) against DTA were established. Investigations showed that both McAbs were characterized as IgG1 with titers of 1: 10(6). The binding of the McAbs to DTA was competitively inhibited by horse sera against DT. The fact that anti-DTA McAbs could be used in western blot analysis and affinity chromatography purification of DT-based immunotoxins implied that they will be useful agents in the studies on DT-based immunotoxins.

Animals↗

PubMed2003

Spatial weak-light solitons in an electromagnetically induced nonlinear waveguide.

We show that a weak probe light beam can form spatial solitons in an electromagnetically induced transparency (EIT) medium composed of four-level atoms and a coupling light field. We find that the coupling light beam can induce a highly controllable nonlinear waveguide and exert very strong effects on the dynamical behavior of the solitons. Hence, in the EIT medium, it is not only possible to produce spatial solitons at very low light intensities but also simultaneously control these solitons by using the coupling-light-induced nonlinear waveguide.

Journal Article↗

PubMed2003

[Chlamydia-like and coronavirus-like agents found in dead cases of atypical pneumonia by electron microscopy].

OBJECTIVE: To explore the causative agents of the atypical pneumonia (also SARS) occurred recently in some regions of our country. METHOD: Organ samples of 7 dead cases of SARS were collected from Guangdong, Shanxi, Sichuan Provinces and Beijing for electron microscopic examination. 293 cell line was inoculated with the materials derived from the lungs to isolate causative agent(s). The agents in the organs and cell cultures were revealed by immunoassay. RESULTS: Both Chlamydia-like and coronavirus-like particles were found in EM. Inclusion bodies containing elementary bodies, reticulate antibodies and intermediate bodies of Chlamydia-like agent were visualized in multiple organs from the 7 dead cases, including lungs (7 cases), spleens (2 cases), livers (2 cases), kidneys (3 cases) and lymph nodes (1 cases), by ultrathin section electron microscopy (EM). In some few sections, coronavirus-like particles were concurrently seen. A coronavirus RNA- polymerase segment (440 bp) was amplified from the lung tissues of two cases of the SARS. After inoculated with materials from the lung samples, the similar Chlamydia-like particles were also found in the inoculated 293 cells. Since the Chlamydia-like agents visualized in both organs and cell cultures could not react with the genus specific antibodies against Chlamydia and monoclonal antibodies against C. pneumoniae and C. psittaci, the results might well be suggestive of a novel Chlamydia-like agent. CONCLUSION: Since the novel Chlamydia-like agent was found co-existing with a coronavirus-like agent in the dead cases of SARS, it looks most likely that both the agents play some roles in the disease. At the present time, however, one can hardly determining how did these agents interact each other synergetically, or one follows another, need further study.

Chlamydia↗

PubMed2003

"Bacillus hackensackii" sp. nov., a novel carbon dioxide sensitive bacterium isolated from blood culture.

An endospore-forming, gram-positive bacillus was isolated from a patient's blood culture. This bacillus did not grow in the presence of 5% carbon dioxide although it grew well in ambient air at 37 degrees C. Although the organism thus is an aerobic bacterium, its sensitivity to increased carbon dioxide concentration places it in a distinct category of gaseous atmospheric requirement: capnophobic. Based on its morphology, growth characteristics, biochemical reactions and a complete 16S rRNA gene nucleotide sequence analysis, this microorganism represents a novel Bacillus species. The clinical significance of this isolate is unknown. It is proposed that the bacterium be classified in the genus Bacillus as "Bacillus hackensackii".

Adult↗