[Clinical features and CT findings of fibrosing mediastinitis associated pulmonary hypertension].

Objective: To investigate the clinical features and CT findings of pulmonary hypertension (PH) in patients with fibrosing mediastinitis (FM). Methods: Thirteen patients with FM diagnosed between September 2015 and June 2022 were studied retrospectively, including patients with PH (FM-PH group) and patients without PH (FM group) confirmed on right heart catheterization. The t test of two independent samples, Mann-Whitney U rank sum and Fisher's test were used to compare the general information, symptoms, laboratory examination, right ventricular and pulmonary artery measurement data and pulmonary artery CT findings between the two groups, respectively. Results: Compared with the 7 FM patients aged 28-79 (60.00±17.69) years, the 6 patients in the FM-PH group, aged from 60 to 82 (68.83±8.35) years, had more peripheral edema, lower percentage of PaO2, wider inner diameters of pulmonary artery and right ventricle, a higher ratio of right ventricle and left ventricular transverse diameter, faster tricuspid regurgitation velocity and higher estimated systolic pulmonary artery pressure (P<0.05). There were no differences in BNP levels and tricuspid annular plane systolic excursion between groups (P>0.05). Of the 6 patients with PH, 5 had precapillary PH and 1 had mixed PH. Except that the pulmonary vascular resistance in patients of the FM-PH group was significantly higher than that in the FM group (P<0.05), there were no significant differences in cardiac output, mixed venous oxygen saturation and pulmonary capillary wedge pressure between the two groups. CT pulmonary angiography (CTPA) showed pulmonary artery and vein stenosis. Patients in the FM-PH group had more severe stenosis and occlusion of pulmonary artery and pulmonary vein (P<0.05), and more involvement of multiple pulmonary veins (P<0.05). Conclusions: The clinical manifestation of FM complicated with PH is related to the degree of involvement of pulmonary artery, vein and airway. It is recommended that the disease be evaluated in combination with multiple parameters such as clinical manifestations, cardiac ultrasound, right cardiac catheter and CTPA.

[A randomized controlled study of peramivir, oseltamivir and placebo in patients with mild influenza].

Objectives: To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza. Methods: This was a randomized, double-blind, double-dummy, placebo and positive control, multicenter clinical trial, comparing peramivir trihydrate with oseltamivir and placebo. The inclusive criteria were 15-70 years old, onset within 48 h, positive rapid influenza antigen test, and febrile (>38℃) accompanied with at least two associated symptoms. The severe cases complicated with chronic pulmonary and cardiac diseases, malignancies, organ transplantation, hemodialysis, uncontrolled diabetes, immunocompromised status, pregnancy and coexistence of bacterium infections were excluded. All patients were randomized 2∶2∶1 to receive peramivir, oseltamivir and placebo respectively. The primary endpoint was the disease duration, the secondary endpoints included time to normal axillary temperature and normal living activities, viral response, and adverse effects. Results: Following informed consent, 133 patients were included in this study. Four patients were exclude due to missing medical records, not fitting inclusion or exclusion criteria and poor compliance. A total of 129 patients were finally analyzed, including 49 cases, 54 cases and 26 cases in peramivir group, oseltamivir group and placebo group. The median disease duration were 96 (76, 120) hours, 105 (90,124) hours, and 124 (104, 172) hours in three groups respectively (P>0.05) . The time to normal axillary temperature, normal living activities and viral response were not significantly different in three groups (P>0.05) . Conclusion: The value of antiviral therapy in patients with mild influenza needs to be further determined.

[Comparison of ultrasound-guided serratus anterior plane block and erector spinae plane blockperioperatively in radical mastectomy].

Objective: To compare the perioperative effects of ultrasound-guided serratus anterior plane block (SAPB) and erector spinae plane block (ESPB) in radical mastectomy. Methods: One hundred and fifty patients,undergoing radical mastectomy from May 2016 to Jan 2019,the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, were randomly divided into SAPB group, ESPB group and control group. Patients in SAPB group and ESPB group were received corresponding blocks before induction of general anesthesia. The control group was only received routine general anesthesia without any block. Patient-controlled intravenous analgesia (PCIA) was performed in all the patients postoperatively. The VAS score at rest or coughing and Ramsay score at 2, 4, 8, 12, 24, 48 h after operation were compared among the three groups. The intraoperative dosages of propofol and remifentanil,press times and sufentanil cumulative dosage of PCIA in 48 hours after operation, postoperative rehabilitation indicators and adverse effects were all compared. Results: In all the three groups,the VAS scores at rest and coughing increased first and then decreased 2 h to 48 h after operation. The VAS scores in SAPB group and ESPB group were lower than that in control group (P<0.05), whereas, no significant difference was observed between SAPB group and ESPB group (P>0.05). For Ramsay score, among the three groups, there were no significances of the main effects of group and time point, as well as interaction effect (all P>0.05). The intraoperative dosages of propofol and remifentanil in SAPB group and ESPB group were lower than those in control group (P<0.05), the press times and sufentanil cumulative dosage of PCIA after operation were also lower than those in control group (P<0.05). There was no significant difference in feeding time after operation among the three groups (P>0.05). The times of first anal exhaust, ambulation and hospitalization after operation in ESPB group and SAPB group were significantly shorter than those in control group (P<0.05). However, there was no significant difference between ESPB group and SAPB group in postoperative rehabilitation indicators mentioned above (P>0.05). The incidences of skin itching and nausea in ESPB and SAPB groups were lower than those in control group (P<0.05). There was no difference in the incidence of vomiting among the three groups (P>0.05). Conclusions: Both SAPB and ESPB can provide good and safe analgesia for radical mastectomy,with equivalent performances in analgesia and adverse effect.

[Effect of obstructive sleep apnea on the severity of acute pulmonary thromboembolism].

Objective: To investigate the effect of obstructive sleep apnea (OSA) on the severity of acute pulmonary thromboembolism (PTE). Methods: Clinical data of patients with acute PTE and OSA who were admitted to Anzhen Hospital from January 2015 to December 2017 were retrospectively analyzed in this study.According to the apnea hypopnea index (AHI), patients were divided into AHI≤15/h group and AHI>15/h group. The levels of oxygen desaturation index (ODI), mean pulse oxygen saturation (MSO(2)), the lowest pulse oxygen saturation (LSaO(2)), oxygen saturation<90% time ratio (Ts90%), hemoglobin, hematocrit (HCT), blood platelet, mean platelet volume (MPV), platelet distribution width (PDW), hypersensitive C-reactive protein (hsCRP), homocysteine (HCY), prothrombin time, activated partial thromboplastin time (APTT), fibrinogen (FIB), D-dimer, fibrin degredation product (FDP), B-type natriuretic peptide, cardiac troponin I (cTnI), arterial partial pressure of oxygen (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)) were compared between AHI≤15/h group and AHI>15/h group. Differences in the severity of PTE between the two groups were compared; binary Logistic regression model was used to analyze the risk factors of acute PTE severity. Results: A total of 75 patients with acute PTE and OSA were enrolled. Patients in the AHI>15/h group were significantly older [(68.7±9.9) vs (62.8±12.8) years old, P=0.029], and were more likely to have chronic heart failure and/or lung disease (53.1% vs 23.1%, P=0.012) and elevated Pulmonary Embolism Severity Index scores (61.2% vs 30.8%, P=0.012) than those in the AHI≤15/h group. Compared to the AHI>15/h group, the levels of D-dimer and B-type natriuretic peptide were much lower in the AHI ≤15/h group [243.0 (140.0, 471.5) vs 408.0 (258.0, 1 009.5) µg/L, P=0.01; 48.0 (19.8, 87.5) vs 70.0 (34.5, 201.0) ng/L, P=0.039], while arterial oxygen partial pressure was significantly higher [(79.0±10.6) vs (73.4±8.2) mmHg (1 mmHg=0.133 kPa), P=0.015]. In patients with acute PTE and OSA, AHI (P=0.030) and B-type natriuretic peptide level (P=0.023) were independently associated with an increased risk of acute PTE severity. Conclusion: In patients with PTE and OSA, moderate or severe OSA may aggravate the severity of acute PTE.

[Application of the BACspreader™ Microbe Dispersion Counter in drug susceptibility test on Mycobacterium tuberculosis].

Objective: To evaluate the application of the BACspreader™ Microbe Dispersion Counter in drug susceptibility test (DST) on Mycobacterium tuberculosis (MTB). Methods: The MTB strains were dispersed and diluted to 1.0 McFarland standard turbidity, by means of BACspreader™ Microbe Dispersion Counter and manual grinding method, respectively. The bacterial dispersion effect and bacterial activity were tested by microscope and colony counting method. During Jan. 2015 to June 2015, a total of 726 isolates of MTB were collected in all district tuberculosis hospitals of Shanghai. The bacterial suspension dispersed by instrument and manual grinding, were inoculated in slant medium for DST (Proportion Method), and then incubated in 37 ℃ incubator for 28 days and the DST results were reported. The effects of the 2 different bacterial dispersion methods were compared by comparing DST results and counting the bacterial colony which grew in high and low concentration control media. Paired chi-square test was used for statistical analysis, and the significance level was 0.05. Results: Compared to the manual grinding method, the MTB colony could be better dispersed by BACspreader™ Microbe Dispersion Counter, without reducing the bacterial activity. The DST results of 726 mycobacterial isolates were the same by different bacterial dispersion methods. The count of bacterial colony growing in high concentration control medium was significantly different between of the 2 dispersion methods (χ(2)=8.0, P<0.01). When counting the bacterial colony growing in low concentration control medium, the numerable rate was 97.7% by BACspreader™ Microbe Dispersion Counter, and 4.3% by manual grinding method; the difference being significant between the 2 dispersion methods (χ(2)=674, P<0.001). Conclusions: Compared to the manual grinding method, the slant medium inoculated with bacterial suspension obtained by BACspreader™ Microbe Dispersion Counter had better countability in low concentration control slants, and had more significant contrast between high and low concentration control slants, which was useful to determine the DST results. Introducing the BACspreader™ Microbe Dispersion Counter to MTB DST could automate the DST process, make the testing results objective and standardized, and reduce personal error in the tests.

[Lung function influenced the prognosis of cardiac surgery in patients with chronic obstructive pulmonary disease].

Objective: To investigate the value of lung function for the prognosis of cardiac surgery in patients with chronic obstructive pulmonary disease(COPD). Methods: A retrospective analysis was conducted on the hospitalized patients with coronary heart disease or valvular heart disease in Beijing Anzhen hospital during Janury 2013 to December 2015. The relationship between lung function and extubation time, time of staying in ICU, second time tracheal incubation, tracheotomy and mortality rate were analyzed. Results: There were 355 patients without surgery in a total 1 729 of patients, of which 65 (18.31%)suffered from severe pulmonary dysfunction. There were 242 patients with obstructive ventilation dysfunction, 75 with mild, 127 with moderate, and 40 with severe and very severe obstructive ventilation dysfunction. There were significant differences in the rates of planned extubation and late extubation between patients with normal lung function (81.6% and 10.7%) and those with abnormal lung function(74.4% and 12.8%). In patients with different GOLD classification (Ⅰ, Ⅱ, and Ⅲ-Ⅳ), there were significant differences in the rates of early extubation 14.7%, 14.2% and 5%, planned extubation (80%, 74% and 65%) and late extubation (5.3%, 11.8% and 30%). By comparison with patients without COPD, the extubation time of COPD patients increased by 20.3%, the time of staying in ICU prolonged by 14.2%, and the mortality rate increased to 180%(P<0.05). However, there was no significant difference in the rate of second time tracheal incubation or tracheotomy. Conclusion: There was a close relationship between GOLD classification and prognosis of cardiac surgery patients with COPD. Cardiac surgery needs to be cautious because of increase in incidence of postoperative adverse events in patients with severe pulmonary dysfunction.

[The clinical feature and microbiological etiology of patients with infective endocarditis].

Objective: To investigate the clinical features, pathogenic distribution and drug susceptibility of patients with infective endocarditis (IE). Methods: Clinical data of IE patients were collected, who were admitted to Capital Medical University Affiliated Beijing Anzhen Hospital from January 2012 to March 2015. Results: Three hundred and three IE patients were enrolled with age of (43±16) years old. Fever (85.5%)and cardiac murmur (62.4%)were the most common clinical presentations. Congenital heart diseases was the leading underlying diseases in IE patients. Non rheumatic valve diseases (13.5%) followed. Vegetations were found in 90.4%(274/303) patients. Streptococcus which accounted for 44.2% was the major pathogen of IE. Staphylococcus (28.9%)was the second common pathogen. Gram-negative bacteria were diversified in categories. Gram-positive cocci were consistently sensitive to vancomycin and linezolid. Conclusions: Congenital heart disease is the main underlying disease related to IE. Streptococcus is still the primary pathogen. Gram-positive cocci keep good sensitivity to vancomycin and linezolid.

A putative miR172-targeted CeAPETALA2-like gene is involved in floral patterning regulation of the orchid Cymbidium ensifolium.

APETALA2 plays critical roles in establishing meristem and organ identity during plant floral development. In this study, we obtained a CeAP2-like gene by using the mRNA differential display technique to analyze the wild type and a multitepal mutant of the orchid Cymbidium ensifolium. The full-length cDNA encoding the CeAP2-like transcription factor shows significant similarity to the cDNA of AP2 from Erycina pusilla and contains nucleotides complementary to miR172. Using a transient gene expression system of Arabidopsis protoplasts, we found that the accumulation of CeAP2-like protein and transcripts was negatively regulated by miR172, indicating this gene as a putative target of miR172. Northern blotting revealed that CeAP2-like is dominantly expressed in the sepals and petals of the wild-type flower, and shows low expression in the gynostemium. In contrast, the accumulation of CeAP2-like transcripts decreased significantly, especially in the central part of the mutant flower, corresponding to its abnormal petals and the absence of the gynostemium. Furthermore, we found an antagonistic expression pattern between CeAP2-like and AGAMOUS in the wild type, representing A- and C-class genes that specify floral organ fate. However, this antagonistic distribution was modified in the multitepal mutant, and both genes showed lower expression than that in the wild type. This result suggested that the balance between CeAP2-like and AGAMOUS activity was important for the regulation of floral patterning in C. ensifolium. This study represents the first report on a class A gene and its regulatory role for floral development in the orchid C. ensifolium.

Molecular characterization and functional analysis of a Flowering locus T homolog gene from a Phalaenopsis orchid.

Warm day and cool night conditions significantly induce reproductive spike formation in Phalaenopsis plants; hence, determining the flowering mechanism regulating the reproductive transition is important. Flowering locus T (FT) plays important roles in flowering induction in several plants. To explore spike induction by warm days and cool nights in Phalaenopsis orchids, we isolated the FT (PhFT) from Phalaenopsis hybrid Fortune Saltzman. The cDNA of PhFT was 809-bp long and contained a 531-bp open reading frame encoding a putative protein of 176 amino acids, a 58-bp 5'-untranslated region (UTR), and a 220-bp 3'-UTR. The predicted molecular mass of PhFT was 19.80 kDa, with an isoelectric point of 8.68. The PhFT was predicted to possess the conserved functional regions of the phosphatidylethanolamine-binding protein superfamily. Nucleotide sequence data indicated that PhFT contained 3 introns and 4 exons. Sequence alignment and phylogenetic analyses of PhFT revealed high homology to the FT proteins of Cymbidium goeringii and Oncidium Gower Ramsey. Quantitative real-time polymerase chain reaction analysis indicated that PhFT mRNA was expressed in roots, apical leaves, mature leaves, and flowers. In flowers, PhFT was expressed more in developing floral buds than in mature flowers and was predominantly expressed in ovaries and petals. Ectopic expression of PhFT in Arabidopsis ft-1 mutants showed novel early-flowering phenotypes that lost their siliques. Our results indicated that the ectopic expression of PhFT could partially complement the late flowering defect in transgenic Arabidopsis ft-1 mutants. Our findings suggest that PhFT is a putative FT homolog in Phalaenopsis plants that regulates flowering transition.

Identification of warm day and cool night conditions induced flowering-related genes in a Phalaenopsis orchid hybrid by suppression subtractive hybridization.

The influence of warm day and cool night conditions on induction of spikes in Phalaenopsis orchids has been studied with respect to photosynthetic efficiency, metabolic cycles and physiology. However, molecular events involved in spike emergence induced by warm day and cool night conditions are not clearly understood. We examined gene expression induced by warm day and cool night conditions in the Phalaenopsis hybrid Fortune Saltzman through suppression subtractive hybridization, which allowed identification of flowering-related genes in warm day and cool night conditions in spikes and leaves at vegetative phase grown under warm daily temperatures. In total, 450 presumably regulated expressed sequence tags (ESTs) were identified and classified into functional categories, including metabolism, development, transcription factor, signal transduction, transportation, cell defense, and stress. Furthermore, database comparisons revealed a notable number of Phalaenopsis hybrid Fortune Saltzman ESTs that matched genes with unknown function. The expression profiles of 24 genes (from different functional categories) have been confirmed by quantitative real-time PCR in induced spikes and juvenile apical leaves. The results of the real-time PCR showed that, compared to the vegetative apical leaves, the transcripts of genes encoding flowering locus T, AP1, AP2, KNOX1, knotted1-like homeobox protein, R2R3-like MYB, adenosine kinase 2, S-adenosylmethionine synthetase, dihydroflavonol 4-reductase, and naringenin 3-dioxygenase accumulated significantly higher levels, and genes encoding FCA, retrotransposon protein Ty3 and C3HC4-type RING finger protein accumulated remarkably lower levels in spikes of early developmental stages. These results suggested that the genes of two expression changing trends may play positive and negative roles in the early floral transition of Phalaenopsis orchids. In conclusion, spikes induced by warm day and cool night conditions were complex in Phalaenopsis orchids; nevertheless, several molecular flowering pathway-related genes were found. The acquired data form the basis for a molecular understanding of spike induction by warm day and cool night conditions in Phalaenopsis orchids.

Role of microglia in the pathogenesis of sepsis-associated encephalopathy.

Sepsis-associated encephalopathy (SAE) is a neurological dysfunction induced by sepsis, which is associated with high morbidity and mortality. However, at present, the cellular and molecular mechanisms of SAE have remained elusive. The pathogenesis of SAE is complex and multifactorial, in which activated inflammation is recognized as a major factor. Pathological characteristics of SAE include blood- brain barrier (BBB) disruption, reduction of cerebral blood fluid (CBF) and glucose uptake, inflammatory response and activation of microglia and astrocytes. The BBB disruption induces the leakage of immune cells and inflammatory mediators, which trigger an inflammatory response in the brain. Inflammatory mediators released by activated microglia and astrocytes cause neuronal loss and brain function defect. In the review we describe the most recent findings in the pathogenesis of SAE and focus on summarizing the major mechanisms related to SAE pathogenesis.

Molecular cloning and characterization of two novel NAC genes from Mikania micrantha (Asteraceae).

NAC proteins, which are plant-specific transcription factors, have been identified to play important roles in plant response to stresses and in plant development. The full-length cDNAs that encode 2 putative NAC proteins, designated as MmATAF1 and MmNAP, respectively, were cloned from Mikania micrantha by rapid amplification of cDNA ends. The full-length cDNAs of MmATAF1 and MmNAP were 1329 and 1072 bp, respectively, and they encoded deduced proteins of 260- and 278-amino acid residues, respectively. The proteins MmATAF1 and MmNAP had a calculated molecular mass of 29.81 and 32.55 kDa and a theoretical isoelectric point of 7.08 and 9.00, respectively. Nucleotide sequence data indicated that both MmATAF1 and MmNAP contained 2 introns and 3 exons and that they shared a conserved genomic organization. Multiple sequence alignments showed that MmATAF1 showed high sequence identity with ATAF1 of Arabidopsis thaliana (61%) and that MmNAP showed high sequence identity with NAP of A. thaliana (67%) and CitNAC of Citrus sinensis Osbeck (62%). Phylogenetic analysis showed that the predicted MmATAF1 and MmNAP proteins were classified into the ATAF and NAP subgroups, respectively. Transient expression analysis of onion epidermal cells indicated nuclear localization of both MmATAF1-GFP and MmNAP-GFP fusion proteins. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis indicated that MmATAF1 was expressed in all the tissues tested, but in varying abundance, while MmNAP was specifically expressed in stems, petioles, shoots, and leaves, but not in roots. The transcript levels of MmATAF1 and MmNAP in shoots and in infected stems were induced and strengthened by wounding, exogenous ZnSO(4), abscisic acid, salicylic acid, and Cuscuta campestris infection on the basis of semi-quantitative RT-PCR and real-time PCR analyses, respectively. Collectively, these results indicated that MmATAF1 and MmNAP, besides having roles in M. micrantha adaptation to C. campestris infection and abiotic stresses, also integrated signals derived from both C. campestris infection and abiotic stresses.

DNA sequence and comparative analysis of chimpanzee chromosome 22.

Human-chimpanzee comparative genome research is essential for narrowing down genetic changes involved in the acquisition of unique human features, such as highly developed cognitive functions, bipedalism or the use of complex language. Here, we report the high-quality DNA sequence of 33.3 megabases of chimpanzee chromosome 22. By comparing the whole sequence with the human counterpart, chromosome 21, we found that 1.44% of the chromosome consists of single-base substitutions in addition to nearly 68,000 insertions or deletions. These differences are sufficient to generate changes in most of the proteins. Indeed, 83% of the 231 coding sequences, including functionally important genes, show differences at the amino acid sequence level. Furthermore, we demonstrate different expansion of particular subfamilies of retrotransposons between the lineages, suggesting different impacts of retrotranspositions on human and chimpanzee evolution. The genomic changes after speciation and their biological consequences seem more complex than originally hypothesized.

Combined surfactant therapy and inhaled nitric oxide in rabbits with oleic acid-induced acute respiratory distress syndrome.

Intratracheal administration of surfactant and inhaled nitric oxide (INO) have had variable effects in clinical trials on patients with acute respiratory distress syndrome (ARDS). We hypothesized that combined treatment with exogenous surfactant and INO may have effects in experimental ARDS. After intravenous infusion of oleic acid in adult rabbits and 4-6 h of ventilation, there was more than a 40% reduction in both dynamic compliance (Cdyn) of the respiratory system and functional residual capacity (FRC), a 50% increment of respiratory resistance (Rrs), a 70% reduction in PaO2 /FIO2, and an increase in intrapulmonary shunting (Q S/Q T) from 4.4 to 33.5%. The animals were then allocated to groups receiving (1) neither surfactant nor INO (control), (2) 100 mg/kg of surfactant (S) administered intratracheally, (3) 20 ppm INO (NO), or (4) 100 mg/kg of surfactant and 20 ppm INO (SNO), and subsequently ventilated for 6 h. After the period of ventilation, the animal lungs were used for analysis of disaturated phosphatidylcholine (DSPC) and total proteins (TP) in bronchoalveolar lavage fluid (BALF), and for determination of alveolar volume density (VV). The animals in the control group had the lowest survival rate, and no improvement in lung mechanics and blood oxygenation, whereas those in the S group had a modest but statistically significant improvement in Cdyn, Rrs, PaO2 and FRC, reduced Q S/Q T, lowered minimum surface tension (gammamin) of BALF, and increased DSPC/ TP and alveolar VV. The NO group had increased PaO2 and reduced Q S/Q T. The SNO group showed improved Cdyn, Rrs, FRC, DSPC/TP, alveolar VV, and gammamin of BALF comparable to the S group, but there was a further increase in survival rate and PaO2, and additional reduction in Q S/Q T and TP in BALF. These results indicate that, in this animal model of ARDS, a combination of surfactant therapy and INO is more effective than either treatment alone.

Serum and egg yolk IgG antibody titers from laying chickens vaccinated with Pasteurella multocida.

Through determining the serum and egg yolk antibody titers in immunized laying hens to Pasteurella multocida regularly, the growth-decline trend of the egg yolk antibody levels was found to be similar to that of the serum antibody levels (r = 0.94), but the growth and decline of the egg yolk antibody seemed to be delayed 3-6 days compared with that of the serum antibody, and the egg yolk antibody titers were generally lower than those of the serum antibody (P < 0.01). Serum and egg yolk antibody levels declined 3 and 6 days, respectively, after booster immunizations. The higher the antibody levels were before booster immunization, the more they declined.

Endogenous substance P mediates cold water stress-induced increase in interleukin-6 secretion from peritoneal macrophages.

Previous studies from this laboratory had shown that exposure of mice to cold water stress leads to an increase in the secretion of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF alpha) from their peritoneal macrophages. We now report that the secretion of IL-6 from peritoneal macrophages is also increased after cold water stress and that the peptide substance P (SP) participates in this stress-induced response. The stress paradigm involved subjecting male C57BL/6J mice to 5 min swim tests in 10 +/- 2 degrees C water twice daily for 4 d. Cold water stress augments the lipopolysaccharide-induced IL-6 secretion from peritoneal macrophages, elevates immunoreactive SP (iSP) in the peritoneal wash fluid, and reduces iSP in certain peritoneum-containing tissues or organs (i.e., diaphragm, abdominal wall, ileum, and rectum). The 10 d stress time studies indicate that increased IL-6 secretion is positively related to elevated iSP in the peritoneal wash fluid and inversely related to reduced iSP in certain peritoneum-containing tissues. Pretreatment with capsaicin, which depletes SP in the sensory nerve endings, eliminates stress-control differences in the peritoneal wash fluid and in certain peritoneal tissues. Moreover, RP67,580, a specific SP antagonist, eliminates the cold water stress-induced augmentation of IL-6 secretion from peritoneal macrophages. These results suggest that cold water stress promotes the release of SP from peritoneal tissues into the peritoneal cavity, where it participates in the cold water stress-induced macrophage functional alterations.

Substance P and stress-induced changes in macrophages.

The present paper further links nervous-endocrine-immune systems by describing influences of SP on the immune system, and more specifically, on macrophage function. We have discussed how macrophages are important to immune responses in that much of cellular and humoral responses depend on macrophage function. Macrophages are sensitive to stress in that cold-water stress causes increased cytokine production, either spontaneously (IL-1), or after induction with LPS (IL-6, TNF alpha). Increased cytokine levels (IL-1, IL-6) may induce acute phase reactants in the liver, which is presumably the mechanism operative in the studies indicating increases in acute phase reactants after certain stressors in animals. SP is a likely candidate to affect immune function. Previous data show that macrophages from various species have receptors for and respond to SP in vitro. SP stimulates phagocytic and chemotactic capacity, as well as increased cytokine, PGE2, and thromboxane B2 production. SP is also involved in neurogenic inflammation and is likely to be involved in the pathogenesis of several inflammatory diseases. Present data indicate SP's involvement in macrophage responses to stress. We have shown that stress induced differential SP receptor binding to peritoneal macrophages, although the precise nature of binding differences has not yet been clearly elucidated. Stress also induces more immunoreactive SP in the peritoneal fluid that bathes the peritoneal macrophages. We hypothesize that the two events, altered SP binding and concomitant increased ligand, are causally related. In addition to other correlational data showing concomitant increased SP binding plus ligand concentrations, there is more direct evidence that SP ligand may induce SP receptor expression since the SP antagonist, CP-96,345, prevents the induction of SP receptor mRNA in the staphylococcal toxin A-induced gastroenteritis (C. Pothoulakis and S. E. Leeman, personal communication). Further supporting our notion for a causal relationship we have found the elimination of SP in vivo (via capsaicin pretreatment) reduced SP binding, as has been previously reported. We have also examined the role of SP on stress-induced altered macrophage function in vitro. SP greatly enhanced the LPS-induced macrophage TNF alpha production from stressed animals; in contrast, it produced relatively little effect on macrophages from control animals. Capsaicin pretreatment diminished the enhanced cytokine production in response to stress, such that levels of TNF alpha and IL-6 approximated those of control mice. Taken together, past and present data suggest that (1) stress may initiate, or at least contribute to, an inflammatory response, and that (2) SP is involved in the macrophage stress response. SP has long been known to be involved in inflammatory processes; our data further suggest its role in mediating stress-induced cytokine alterations.

Further evaluation of one-point microcapsule agglutination test for diagnosis of leptospirosis.

A total of 100 serum samples including 22 acute phase sera and 39 paired sera collected from clinically diagnosed cases of leptospirosis in Ming-shan County, Sichuan Province, China were examined by the one-point microcapsule agglutination test (MCAT), which was developed in Japan, and by conventional microscopic agglutination tests (MAT). The one-point MCAT is more reactive to IgM antibody than MAT and is superior in detecting antibodies in the early stages of the disease.