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Objective: To establish a method of isolate, purify, primary culture and identify human alveolar type II cells (AT II ) in vitro, as well as its possible maintaining phenotype characteristics., Methods: The marginal lung tissue was collected. AT II cells were isolated with trypsin and elastase, purified by a series of steps, such as, cell sieve filtration, differential adhesion, gradient separation and anti-CD14 beads separation. AT II cells were identified with immunofluorescence of human pro-surfactant-associated protein C (pro-SP-C), Green DND-26 probe and electron microscope. The purity of AT II cells was measured by immunofluorescence of human pro-SP-C and Green DND-26 probe. The viability of AT II cells was measured by trypan blue staining. The phenotypes (SP-A, SP-B,SP-C, SP-D) were monitored with reverse transcription-polymerase chain reaction (RT-PCR) at different time points., Results: The output of AT II cells from lung tissue was (5-10) x 105/g, and the cell viability was (93 ± 2)% with trypan blue staining, the cell purity was about 98% with pro-SP-C immunofluorescence and Green DND-26 fluorescent probe, the lamellar bodies were clearly observed with transmission electron microscope. In the aspect of phenotypes maintaining, the time of surfactant expression was about 24 days [SP-A: 0.52 + 0.03 (day 16), 0.35 + 0.02 (day 20),0.26 ± 0.01 (day 24), 0.10 + 0.08 (day 28); SP-C: 0.68 0.16 (day l6), 0.31 + 0.04 (day 20), 0.18 + 0.06 (day 24), 0.14 + 0.09 (day 28)], and the longest one was more than 28 days [SP-B: 1.05 + 0.17 (day 16), 0.76 + 0.35(day 20), 0.55 0.15 (day 24), 0.36 0.19 (day 28); SP-D: 0.52 0.19 (day 16), 0.33 + 0.12 (day 20), 0.31 +0.04 (day 24), 0.23 ± 0.02 (day 28))., Conclusion: We successfully established a procedure to separate, purify,identify of AT II cells, which retain primary phenotypic characteristics over long period.

Objective: To investigate the value of intrathoracic blood volume index (ITBVI) monitoring in fluid management strategy in septic shock patients., Methods: In a prospective study, 33 patients who were diagnosed to be suffering from septic shock in the intensive care unit (ICU) were enrolled . Seventeen patients who received pulse indicator continuous cardiac output (PiCCO) monitoring, and ITBVI was used as indicator of fluid management, were enrolled into ITBVI group; 16 patients who received traditional fluid management strategy [directed by central venous pressure (CVP)] were enrolled into control group. Acute physiology and chronic health evaluation II (APACHEII) score, sepsis related organ failure assessment (SOFA) score and vasopressor score were compared between 1 day and 3 days of treatment. The characteristics of fluid management were recorded and compared within 72 hours., Results: (1)In 3 days of treatment, APACHEII, SOFA and vasopressor score were significantly lower in ITBVI group compared with that of in 1 day of treatment[21.3±6.2 vs. 25.4±7.2, 6.1±3.4 vs. 9.0±3.5, 5 (0, 8.0) vs. 20.0 (8.0, 35.0), respectively, all P<0.01], whereas there were no changes in control group. (2)Although fluid output (ml) was higher in ITBVI group during 48-72 hours period (2 421± 868 vs. 1 721±934, P=0.039), there was no difference in fluid intake, fluid output or fluid balance (ml) within 0-72 hours between two groups (fluid intake: 9 918±137 vs. 10 529±1 331, fluid output : 6 035±1 739 vs. 5 827±2 897, fluid balance: 3 882±1 889 vs. 4 703±2 813, allP>0.05). (3)Comparing the fluid volume (ml) used for fluid replacement period, except that there was no significance in fluid challenge with colloid during 0-6 hours between two groups [ml: 250 (125, 500) vs. 250 (69,250), P>0.05], more fluid intake (ml) was found in ITBVI group [0-6 hours crystalloid: 250(150,250) vs. 125 (105,125), 6-72 hours crystalloid: 125 (125, 250) vs. 100 (56, 125), 0-72 hours crystalloid: 250(125, 250) vs. 125 (75, 125), 6-72 hours colloid: 125 (106, 250) vs. 75 (50, 125), 0-72 hours colloid: 200 (125, 250) vs. 100 (50, 125),all P<0.01]., Conclusion: Clinical picture in patients with septic shock is improved after 3 days of treatment than 1 day of treatment under fluid management directed by ITBVI, compared with by CVP. This improvement may be attributable to accurate assessment of preload and appropriate infusion rate in fluid challenge.

Objective: To study the clinical significance of Toll-like receptor 4 (TLR4) in children with severe sepsis., Methods: A prospective control study was performed. All cases were enrolled from pediatric department of the First Affiliated Hospital of Guangzhou Medical College, and they were divided into severe sepsis group (14 patients) who were diagnosed to have severe sepsis or septic shock in intensive care unit (ICU), pneumonia group (10 cases) with diagnosis of bronchial pneumonia, and healthy control group (10 healthy children). Venous blood samples of 2 ml were collected at admission, the level of TLR4 was detected by flow cytometry .At the same time, the changes in serum interleukin (IL-6, IL-10 ) and tumor necrosis factor-α (TNF-α) levels were determined by enzyme linked immunoadsorbent assay (ELISA)., Results: In severe sepsis group, the contents of TLR4 [(71.56±15.32)%], IL-6 [(1.98±1.55) ng/L], IL-10 [(88.20±61.23) ng/L] and TNF-α [(104.08±85.36) ng/L] were significantly higher than those in pneumonia group [(50.07±26.36)%, (0.93±0.16) ng/L, (41.42±7.02) ng/L, (48.96±6.40) ng/L] and healthy control group [(39.43±17.43)%, (0.94±0.43) ng/L,(43.73±22.68) ng/L, ( 49.94± 18.47) ng/L, all P<0.05). But there was no significant difference in the contents of TLR4, IL-6, IL-10 and TNF-α between pneumonia group and healthy control group (all P>0.05)., Conclusion: This study suggests that TLR4 might be critically involved in the development of sepsis, and changes in TLR4 expression are parallel with levels of proinflammatory cytokines, including IL-6, TNF-α, and IL-10. The combination of TLR4 and proinflammatory cytokines would serve as the predictive parameters in early diagnosis and severity evaluation of sepsis in children.

Objective: To look for the natural ligand(s) of human triggering receptor expressed on myeloid cell-1 (TREM-1), in order to provide the theoretical basis for elucidation of the pathogenesis of sepsis., Methods: Neutrophils and monocytes isolated from human peripheral blood were treated with heat-inactivated Staphylococcus aureus, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus L-form or Pseudomonas aeruginosa L-form respectively for 24 hours. The cell wall was extracted from Staphylococcus aureus, Pseudomonas aeruginosa and Mycobacterium tuberculosis by ultrasound. Neutrophils and monocytes were isolated and treated with the cell wall respectively for 24 hours. Neutrophils and monocytes were isolated and treated with three main components from bacterial cell wall (polysaccharides, lipids and proteins) respectively for 24 hours. The level of TREM-1 mRNA was measured with fluorescent quantitative polymerase chain reaction (PCR), and the concentrations of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) were measured with enzyme-linked immunosorbent assay (ELISA)., Results: The TREM-1 mRNA level and the concentrations of TNF-alpha and IL-1 beta in cell supernatant of neutrophils and monocytes were upgraded when treated with cell, cell wall and cell wall polysaccharides of Staphylococcus aureus and Pseudomonas aeruginosa. Compared with the blank control group, the TREM-1 mRNA level of neutrophils and monocytes was upgraded to (3.86+/-0.20)-fold and (5.15+/-0.56)-fold respectively when treated with cell wall polysaccharides of Staphylococcus aureus (both P<0.05); the TREM-1 mRNA level of neutrophils and monocytes was upgraded to (4.03+/-0.15)-fold and (7.22+/-0.73)-fold respectively when treated with cell wall polysaccharides of Pseudomonas aeruginosa (both P<0.05). The effect could be attenuated by the addition of LP17 which could bind TREM-1 ligand. This attenuating effect was not found when the cells were treated with cell, cell wall or cell wall polysaccharides of Mycobacterium tuberculosis., Conclusion: The study provides the evidence that TREM-1 natural ligand(s) is present on cell wall of bacteria including Staphylococcus aureus and Pseudomonas aeruginosa, and it might be polysaccharides.

Objective: To evaluate compliance with bundle treatment in the management of severe infection in a tertiary hospital, aiming at analyzing clinical data in order to popularize guidelines for management of severe sepsis and septic shock., Methods: A 14-month (from November 1, 2006 to December 31, 2007) prospective observational study of a group of 43 patients admitted to the respiratory intensive care unit in First Affiliated Hospital (tertiary hospital) of Guangzhou Medical College meeting the criteria for severe pneumonia and septic shock was carried out. Implementation of 6-hour and 24-hour bundle treatment for severe infection was divided into three phases consisting of education, trial, and application. A cohort of 43 patients with matched disease history admitted during January 1, 2004 to October 31, 2006 were enrolled as control group., Results: (1) In 6-hour bundle treatment for severe infection, 20.9% (9/43) had serum lactate measured, blood culture was obtained prior to antibiotic administration in 7.0% (3/43) of patients, 100% (43/43) had empirical antibiotics administration within 1 hour, an infusion of an initial minimum of 20 ml/kg of crystalloid or colloid equivalent (1.1 ml/kg of 20% albumin or 4.8 ml/kg of 6% hydroxyethyl starch) was given in 44.2% (19/43), with infused fluid (converted into 6% hydroxyethyl starch) reaching (503.95+/-176.19) ml within 6 hours, in 94.7% (18/19) of patients had received vasopressors , and inotropic dobutamine and/or transfusion of packed red blood cells were administered in 7.0% (3/43). (2) In 24-hour bundle treatment for severe infection group, 31.6% (6/19) had received low-dose steroids, 34.9% (15/43) had their blood glucose controlled<8.3 mmol/L, mechanical ventilation with inspiratory plateau pressures maintained<30 cm H(2)O (1 cm H(2)O=0.098 kPa, 6 ml/kg tidal volume) was instituted in 97.6% (40/41) of patients. (3) The percentage of compliance with 6-hour and 24-hour bundle treatment for severe infection were 0 and 21.4% respectively, total compliance was also 0. (4) As compared with control group, a 23.30% absolute mortality reduction was found in bundle group (18.6% vs. 41.9%, P=0.019)., Conclusion: Bundle treatment for severe infection is complied with partially in our hospital, suggesting that it is still quite arduous to popularize guidelines for management of severe sepsis and septic shock in our country.

Objective: To investigate the present situation of general intensive care unit (ICU) in second grade hospitals, and to establish intensive care network for the Department of Public Health of Guangdong province in Guangdong province., Methods: Data from ICU of 26 hospitals in Guangdong were collected through questionnaire concerning different aspects of critical care medicine., Results: (1) ICU size was (10.12+/-3.82) beds per unit, ratios of doctors to beds and nurses to beds were 0.73+/-0.25 and 1.80+/-0.57 respectively, and proportions of closed model or semi-closed model of ICU management were 69.2% and 26.9% respectively. (2) Area occupied by per bed was (17.57+/-7.58) m2, ratio of basins with infrared control facet to beds was 0.47+/-0.33, proportions of ICU equipped with room equipped with positive or negative air pressure, laminar flow, or with room for preparing nutrition support were 15.4%, 30.8%, and 23.1% respectively. (3) All the ICU were capable of institution and management of artificial airway, mechanical ventilation, placement of deep vein line, cardioversion and defibrillation, parenteral nutrition, and sedation. Ninety-six point two percent of the ICU could accomplish trachea intubation independently. Fifty-three point eight percent of the ICU could perform hemodynamic monitoring. Continuous blood purification could be done in 73.1 % of the ICU. (4) Ninety-six point two percent of the ICU were equipped with continuous bedside multifunctional electrocardiogram monitor and ratio of the monitors to beds was 0.89+/-0.29. Ratios of resuscitation air bags to beds and ventilators to beds were 0.71+/-0.34 and 0.71+/-0.24 respectively. Portable ventilator was equipped in 34.6 % of the ICU. Forty percent of the ICU could not perform non-invasive ventilation, 65.4 % of the ICU were equipped with fiberoptic bronchoscope, blood gas analysis could be done during 24 hours round in 92.3 % of the ICU. (5) Twenty-six ICU investigated were found to be distributed over the district of Zhujiang delta, and east, north and west regions of Guangdong, forming the base of intensive care network in Guangdong province., Conclusion: Most of the general ICU in second grade hospitals in Guangdong province have fulfilled the main requirement for ICU in accordance with the guidelines for construction of ICU in Guangdong province and of guidelines for construction and management of ICU in China. The average level of the ICU is close to level II. It is possible for the intensive care network to integrate, the present resources effectively and then enhance the level of treatment of critical illness in the said district.

Objective: To review retrospectively the effect of continuous blood purification (CBP) on septic shock with acute renal failure and respiratory failure as a result of severe pneumonia, and to analyze its relationship with prognosis., Methods: Twenty-five patients diagnosed as severe pneumonia with varying degrees of multiple organ failure (MOF), septic shock and anuria, were allocated into three groups based on outcome of the patients A (7 patients), died of progressive worsening of septic shock, (9 patients, yet died of severe pneumonia afterwards). Laboratory data and critical scored of the patients 1 day before and 24, 48, and 72 hours after the initiation of CBP were compared among the three groups., Results: (1) The survival rate in group A was 28%, while the combined fatality in groups B and C was 72%. (2) CBP was forced to be stopped in group B patients within less than 48 hours from the start of treatment. Single-factor analysis suggested: group B exhibited higher level of blood sugar compared with group A [(13.17+/-5.84) mmol/L vs. (8.07+/-2.28) mmol/L, P<0.05], and higher fibrinogen levels compared with group C[(5.75+/-3.08) g/L vs. (3.10+/-1.06) g/L, P<0.05] before the treatment. Forty-eight hours after the initiation of CBP, patients in group B exhibited higher fibrinogen and dopamine levels compared with those of groups A and C [(8.24+/-3.57) g/L vs. (5.13+/-0.94) g/L, (3.01+/-1.22) g/L, P<0.05 and (12.00+/-6.93) microgxkg(-1)xmin(-1) vs. (1.00+/-2.45) microgxkg(-1)xmin(-1), (2.89+/-4.37) microgxkg(-1)xmin(-1), P<0.05, respectively]. (3) Acute physiology and chronic health evaluation III (APACHE III) score of group A before treatment was significantly lower than those in groups B and C (89.43+/-11.28 vs. 108.00+/-15.10 and 104.67+/-13.77, both P<0.05). After 72 hours of CBP treatment, patients in groups A and C showed significantly different changed in APACHE II scores compared with group B (-10.43+/-4.89, -9.11+/-3.76 vs. -2.33+/-4.39, P<0.05) and APACHE III scores (-2.14+/-2.19, -1.00+/-1.87 vs. 0.56+/-1.88, P<0.05)., Conclusion: (1) CBP is curative for some patients in septic shock with acute renal failure and respiratory failure as a result of severe pneumonia, with and overall survival rate of 28%. (2) APACHE III score is a sensitive index before and after CBP treatment, and scores of 90-100 may be taken as an indication for CBP. (3) High blood sugar and fibrinogen levels may be potential risk factors, in particular, a high fibrinogen level implies a poor prognosis.

Objective: To investigate the changes and pattern of pulmonary function in severe acute respiratory syndrome (SARS) patients during convalescent period., Methods: Pulmonary function tests were performed in 26 SARS convalescent patients regularly every 3 months after their discharge from hospital. The significance of changes in pulmonary function indexes was analyzed., Results: Restrictive pulmonary ventilation function and diffusing dysfunction of the lung were found in one third of the patients during third to sixth month from onset. There was a little improvement in forced vital capacity (FVC), one second forced expiratory volume (FEV(1.0)), functional residual capacity (FRC) and residual volume (RV) as convalescent period was prolonged, but no difference was found between different stages (3-6 months, 6-9 months, 9-12 months, 12-15 months and 15-17 months). Meanwhile, the FEV(1.0)/FVC showed no significant change. However, there was an obvious improvement in total lung capacity (TLC) and diffusing capacity of the lung for carbon monoxide (DLCO) with the elapse of time, and also a significant difference was found between the later stage and the earlier stage., Conclusion: Pulmonary dysfunction is found among some SARS patients after convalescence presenting mainly as restrictive ventilatory function and diffusing capacity abnormality. These dysfunctions would improve gradually with the elapse of time.

Objective: To analyze the laboratory characteristics of patients with sporadic severe acute respiratory syndrome (SARS) in China., Methods: The laboratory findings of the 4 cases with SARS occurring in Guangzhou, People's Republic of China, in 2004 were analyzed and compared with that during epidemic., Results: Leukopenia and lymphocytopenia were seen in all the patients. Two patients had slightly decreased peripheral blood T lymphocyte count. Alanine aminotransferase (ALT) and aspartate transaminase (AST) levels increased slightly in 3 patients. No hypoxemia was seen in all the patients. Both SARS-IgM and IgG sero-conversion occurred earlier in all the patients with the titer increased more than 4-fold shortly. Neutralization test was positive in all the patients. SARS coronary virus (SARS-CoV) RNA was detected by polymerase chain reaction (PCR) in pharyngeal swabs only in 1 patient., Conclusion: The 4 sporadic SARS patients in 2004 have milder manifestations, shorter course of disease with no complications during an epidemic, compared with patients seen previously. The change in laboratory findings is less than that, which might be attributable to milder virulence of the SARS-CoV. The antibody appears earlier in these patients. The SARS-CoV is eliminated rapidly.

Objective: To discuss the possible death risk factors of severe acute respiratory syndrome (SARS) with acute respiratory distress syndrome (ARDS)., Methods: Twenty-five patients suffered from SARS with ARDS in the intensive care unit were retrospectively analyzed from December 2002 to April 2003. Statistical analysis was made using SPSS 10.0 and forward stepwise (wald) logistic regression analysis were used to determine the interrelationships between multiple variables and death. P<0.05 was considered statistically significant., Results: The following factors were associated with a significantly higher mortality rate in the SARS with ARDS patients, including age increase (OR=1.203, CI=1.036 to 1.396, P=0.016), long-time hypoxia(OR=1.067, CI=1.014 to 1.122, P=0.013), thrombocytopenia(OR=111.932, CI=6.096 to 2 055.252, P=0.001), hypernatremia (OR=26.667, CI=2.242 to 317.147, P=0.009), and elevation of serum creatinine levels (OR=111.932, CI=6.096 to 2 055.252, P=0.001)., Conclusion: More attention should be paid to deal with these risk factors and to prevent the development of serious complications associated with SARS.

Objective: To analyze the clinical features and therapy experience of severe acute respiratory syndrome (SARS)., Methods: From December 2002 to April 2003 in Guangzhou Institute of Respiratory Disease, 38 patients with severe SARS were retrospectively studied to evaluate the relationship between treatment strategy and prognosis., Results: Thirty-eight cases of severe SARS were diagnosed. Comprehensive measures most commonly included corticosteroids, antibiotics, antivirotics, nutritional support and mechanical ventilation. Thirty cases were cured (78.9%), of them 11 cases had pulmonary fibrosis (36.7%), 8 patients died (21.1%) in all cases., Conclusion: Severe SARS might develop rapidly. In addition to early diagnosis, prompt isolation, and emergency therapy, appropriate use of corticosteroid and noninvasive ventilation should be recommended.