Predictive values of cervix length measurement based on transvaginal ultrasonography combined with pathological examination of placenta for premature delivery and correlation between premature delivery and infection.

OBJECTIVE: The predictive values of cervix length (CL) measurement based on transvaginal ultrasonography (TVUS) and pathological examination of placenta for premature delivery (PTD) were investigated, and the correlation between PTD and infection was analyzed. PATIENTS AND METHODS: A total of 120 pregnant women with PTD or high-risk factors for PTD admitted to Hengyang Maternal and Child Health Hospital, between February 2020 and March 2022 were included in this retrospective study. There were 36 subjects in the PTD group and 84 in the normal delivery group (control group). They underwent pathological examination of the placenta and TVUS for CL measurement. The final gestational age was set as the standard for the evaluation of the predictive values of pathological examination of the placenta and TVUS. Moreover, a pathological examination of the placenta was used to analyze the correlation between PTD and infection. RESULTS: The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of joint inspection were remarkably superior to those of single CL or pathological examination of the placenta (p<0.05). The proportion of pregnant women with CL ≤30 mm and positive placental pathology was higher than that of pregnant women with CL >30 mm and negative placental pathology (p<0.05). In addition, the incidence of Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), and chorioamnionitis (CA) in the vaginal discharge of the PTD group was markedly superior to that of the control group (p<0.05). CONCLUSIONS: The combination of CL ≤30 mm and positive placental pathology could effectively predict PTD, and placental infection was notably correlated with the occurrence of PTD.

The dietary combination of essential oils and organic acids reduces Salmonella enteritidis in challenged chicks.

This study was conducted to determine the effects of essential oils and organic acids (EOA) on Salmonella Enteritidis (S. Enteritidis) challenged chickens. One-day-old specific pathogen-free (SPF) chicks (250) were randomly assigned to 5 groups, with 50 birds in each group. The treatment groups were as follows: 1) basal diet, negative control group (NC); 2) basal diet + S. Enteritidis, positive control group (PC); 3) PC + 4,000 g/t of enrofloxacin (5%), antibiotic group (ENR); 4) PC + 800 g/t of EOA1, thymol-benzoic acid group (TBA); and 5) PC + 800 g/t of EOA2, cinnamylaldehyde-caproic acid group (CCA). At 7 D of age, each bird, except those in NC, was orally gavaged with 0.4 mL of a suspension of 4.4 × 109 cfu S. Enteritidis/mL. Results revealed that ENR reduced bacterial counts in the liver and spleen on days 3, 5, and 7 post-challenge more (P < 0.05) than any other treatments. However, bacterial counts in cecal contents among ENR, TBA, and CCA were similar at 5 and 7 D post-challenge but lower than those of PC. Additionally, the bacterial counts in liver, spleen, and cecum contents in TBA were lower (P < 0.05) than in PC at 3, 5, and 7 D post-challenge; the bacterial counts in spleen contents in TBA were lower (P < 0.05) than in CCA at 7 D post-challenge. Tumor necrosis factor-α contents in TBA and CCA were lower (P < 0.05) than those in PC. Also, the ratio of villus height to crypt depth in the ileum of CCA was higher (P < 0.05) than that of PC and ENR; however, there was no difference in the secretory IgA content of the jejunum among the groups. In conclusion, EOA had a bacteriostatic effect on S. Enteritidis, and the effect of the thymol-benzoic acid complex surpassed that of the cinnamaldehyde-caproic acid complex. Therefore, EOA may act as an effective antibiotic substitute for animals in the prevention and treatment of Salmonella.

A mixture of daidzein and Chinese herbs increases egg production and eggshell strength as well as blood plasma Ca, P, antioxidative enzymes, and luteinizing hormone levels in post-peak, brown laying hens.

Dietary supplementation with the isoflavone, daidzein, has been shown to improve egg production in poultry. Additionally, providing Chinese herbs (CH) in the broiler diet has led to increased antioxidative enzyme activity. However, the combined effect of these dietary supplements on hen performance has not been examined. Therefore, the objective of this study was to determine if dietary supplementation with a mixture of daidzein and CH would alter laying performance, egg quality, and blood plasma constituents of post-peak laying hens. At 59 wk of age, Hyline brown hens (240) were randomly allocated to 2 dietary groups and fed for 16 wk. The control group received the basal diet, and a treatment group was fed the basal diet that contained 0.02% of a mixture of daidzein and CH. Egg production and weight were recorded daily and egg quality data were collected at 75 wk of age. Blood plasma antioxidant activity, hormone levels, mineral (Ca and P) content, and osteocalcin content were determined at the end of the study. The results showed that laying rate, egg mass, and shell strength were greater in the daidzein-CH mixture group than the controls (P < 0.05). The plasma glutathione peroxidase, superoxide dismutase, and luteinizing hormone levels were also greater in the daidzein-CH mixture group compared with the control group (P < 0.05). The results of this study reveal that supplementing diets with a daidzein-CH mixture can improve laying performance perhaps by increasing plasma antioxidant activity, luteinizing hormone levels, and mineral content.

[Cognitive level of first aid knowledge regarding small area burn among 2 723 child caregivers in Shanghai: a cross-sectional survey and analysis].

Objective: To assess the cognitive level of first aid knowledge regarding the small area burn among the child caregivers in Shanghai and improve the level of first aid for small area burn in children. Methods: From November 2017 to March 2018, 7 municipal districts in Shanghai were selected according to the random number table, from which 2 750 students of 4 nurseries, 5 kindergartens, 6 primary schools, and 2 junior middle schools were selected by adopting the convenience sampling method. Each student was limited to one caregiver as the research object. A cross-sectional survey was conducted on the cognitive level of first aid knowledge regarding small area burn among the caregivers with self-designed questionnaire through WeChat and Tencent QQ. The age, burn experience, and scarring after burns in children, the prevalence rate of burn in children of different age groups, the educational background of caregivers and their social relationship with their children, and the measures taken by caregivers firstly after small area burn occurred among their children were recorded. The choices of applying the folk prescription drugs to the wounds of their children made by caregivers and those with different educational backgrounds were recorded. The choices of applying daily necessities to the wound of their children made by caregivers were recorded. The caregivers' knowledge of standard first aid measures for small area burn, and the knowledge of caregivers with different educational backgrounds of all standard first aid measures for small area burn were recorded. The caregivers' choices of hospitals for treatment the first time, and the choices of going to the Grade Ⅲ Level A hospital with burn specialty for treatment made by caregivers with different knowledge levels about first aid measures for small area burn and those by caregivers whose children did or didn't have burn experience were recorded. The caregivers' choices of different types of medical institutions with burn specialty or specialized in burn treatment, and choices of going to burn department of comprehensive Grade Ⅲ Level A hospital for treatment made by caregivers with different knowledge levels about first aid measures for small area burn were recorded. Data were processed with Pearson chi-square test and partitions of chi-square test. Results: The effective recovery rate of questionnaire was 99.0% (2 723/2 750). The ages of children were mainly 6-11 years [64.7% (1 762/2 723)]The prevalence of burn in children was 19.4% (527/2 723). There was no statistically significant difference in the overall comparison of burn prevalence of children among the age groups (χ(2)=1.424, P>0.05). The percentage of scar formation after burn in children was 27.3% (144/527). The education backgrounds of caregivers were mainly undergraduate [40.2% (1 094/2 723)], and their social relationships with children were mainly children's mothers [74.6% (2 030/2 723)]. Assuming that their children suffered from minor burns, the measures firstly taken by 74.0% (2 016/2 723) of the caregivers was to immediately access cool running water and remove clothing on the wound of children. Totally 19.2% (523/2 723) of the caregivers chose to apply folk prescription drugs for their burn children by themselves, and the percentage of caregivers with education background of junior middle school choosing to apply folk prescription drugs for their burn children by themselves was significantly higher than that of caregivers with education background of junior college, undergraduate, or graduate (χ(2)=18.502, 20.642, 13.319, P<0.05). Totally 49.2% (1 340/2 723) of caregivers chose to daub many kinds of daily necessities for their burn children by themselves. Totally 39.2% (1 068/2 723) of caregivers knew all standard first aid measures for small area burn, the percentage of caregivers with education background of undergraduate knowing all standard first aid measures for small area burn was significantly higher than that of caregivers with education background of senior high school and secondary specialized school (χ(2)=11.234, P<0.05). Assuming that their children suffered from minor burns, 39.0% (1 063/2 723) of the caregivers chose to go to the nearest hospital for treatment the first time, the percentage of caregivers who knew all standard first aid measures for small area burn choosing to go to Grade Ⅲ Level A hospital with burn specialty for treatment the first time was similar with that of caregivers who did not know/did not fully know (χ(2)=3.528, P>0.05), and the percentage of caregivers whose children had burn experience choosing to go to Grade Ⅲ Level A hospital with burn specialty for treatment in the first time was similar with that of caregivers whose children didn't have burn experience (χ(2)=3.521, P>0.05). Among all medical institutions with burn specialty or specialized in burn treatment, 28.0% (762/2 723) of the caregivers chose to go to comprehensive Grade Ⅲ Level A hospital for treatment, and the percentage of caregivers who knew all standard first aid measures for small area burn choosing to go to comprehensive Grade Ⅲ Level A hospital for treatment was significantly higher than that of caregivers who did not know/did not fully know (χ(2)=4.890, P<0.05). Conclusions: The caregivers of children are mainly children's mothers with education background of undergraduate in Shanghai, and caregivers' cognitive levels of first aid knowledge regarding the small area burn are low. Only a few caregivers know all standard first aid measures for small area burn, and there are still some caregivers who have the wrong idea of applying folk prescription drugs or daily necessities for children by themselves. The publicity and education of basic first aid knowledge of burn should be strengthened through various channels such as burn simulation exercise and network, and caregivers should be guided to take their children to hospitals with burn specialty for treatment after occurrence of burn in children, so as to obtain more professional medical treatment.

Genistein increases progesterone secretion by elevating related enzymes in chicken granulosa cells.

Genistein, a biologically active isoflavone, exists in many soy products. It is well known that genistein binds to both oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERß), but it has a higher affinity to ERß. Genistein can also bind to the G protein-coupled receptor 30 (GPR30, also known as G protein-coupled oestrogen receptor 1 or GPER). Furthermore, weak oestrogenic activity has been found in genistein, but the mechanism of action remains unknown. The aim of this study was to investigate the in vitro effects of genistein on the secretion of progesterone (P4) and oestradiol (E2) in chicken granulosa cells harvested from follicles, as well as the mRNA expression of ERs in these cells. In addition, we examined the expression of key enzymes including steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage (P450scc), and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) in the process of P4 synthesis. The results showed that genistein did not affect the viability of granulosa cells, nor was the proliferating cell nuclear antigen (PCNA) protein changed. Among the 1-, 10-, 100-, and 1,000-nM concentrations tested, treatment with 1 nM genistein for 48 h significantly increased P4 but did not affect E2 secretion. Real-time PCR results showed that the ERß gene expression in granulosa cells was markedly upregulated by 1 nM genistein treatment for 48 h, but there was no significant difference in ERα and GPR30 expression. Genistein also increased the gene expression of StAR, P450scc and 3ß-HSD in the cultured granulosa cells. These results indicate that genistein acts directly on chicken granulosa cells to increase P4 production by upregulating the gene expression of key enzymes through binding in ERß. It may exert positive effects on the reproduction of late-laying hens and act as an effective and safe feed additive for animals.

Epidemiology and outcomes of bloodstream infections in 177 severe burn patients from an industrial disaster: a multicentre retrospective study.

OBJECTIVES: To determine the characteristics of bloodstream infections (BSIs) and to evaluate the impact of BSIs on mortality in severe burn patients. METHODS: A retrospective observational study was conducted in 20 tertiary hospitals. A total of 185 patients who experienced a massive dust explosion in eastern China were included. RESULTS: After exclusion, 177 patients were analysed. The median total body surface area (TBSA) burned was 95% (interquartile range 85%-98%). Inhalation injuries occurred in 97.2%. The overall 90-day mortality was 35% (62/177). During the study period, 120 (67.8%) patients developed 253 episodes of BSI with 323 unique causative pathogens. Sixty-six episodes were polymicrobial infections. Catheter-related BSIs (CRBSIs) accounted for 41.5% of the episodes. Acinetobacter baumannii (19.5%), Klebsiella pneumoniae (13.9%) and Candida (12.7%) were the most common organisms. Antimicrobial resistance was found in 63.5% of the isolates, particularly in Gram-negative bacteria. Patients who developed BSIs had a greater illness severity at admission to the intensive care unit, and worse outcomes. After adjusting for demographics, severity of illness and treatment characteristics in a multivariate logistic model, there was a trend toward BSI increasing the risk of 90-day mortality (adjusted OR 3.4; 95% CI 0.9-12.9; p=0.069). In subgroup analyses, CRBSIs (adjusted OR 5.7; 95% CI 1.3-24.9; p=0.021 versus no BSI) and polymicrobial BSIs (adjusted OR 6.1; 95% CI 1.3-28.1; p=0.020 versus no BSI) had greater risk of 90-day mortality. CONCLUSIONS: A strikingly high rate of BSIs was observed in severe burn patients. Gram-negative organisms and fungi were the leading causes. CRBSIs and polymicrobial BSIs were associated with high mortality.

[Relationship between hypertension and body mass index, waist circumference and waist-hip ratio in middle-aged and elderly residents].

Objective: To assess the relationship between hypertension and BMI, waist circumference and waist-hip ratio in middle-aged and elderly residents in Luzhou, Sichuan province. Methods: A total of 2 033 middle-aged and elderly local residents aged 35-69 years were enrolled from Luzhou through stratified cluster sampling from March 27 to April 20, 2015. A face-to-face questionnaire survey and physical examination were conducted by trained investigators. Results: The overall prevalence rate of hypertension was 43.48%. The overweight rate, obesity rate, centrality obesity (calculated according to waist circumference) and centrality obesity (calculated according to waist-hip ratio) were 42.5%, 14.6%, 48.4% and 74.0%, respectively. The multivariate logistic analysis showed that gender and age were related to the prevalence of hypertension. Compared with age group <40 years, the OR values were 2.066 and 4.756 respectively in age groups 45-60 and ≥60 years. After control the confounding effect of gender and age, overweight, obesity and centrality obesity (calculated according to waist circumference) were risk factors for hypertension, waist-hip ratio was not used in the regression equation. BMI and waist circumference or waist-hip ratio had combined effect on the prevalence of hypertension. Compared with the normal adults, the risk for hypertension increased as the increase of the level of overweight and obesity [OR from 1.524 (95%CI: 1.044-2.226) to 4.461 (95%CI: 3.405-6.326) and OR from 1.569 (95%CI: 1.134-2.171) to 5.468 (95%CI: 3.797-7.876)]. Conclusions: The influences of BMI, waist circumference and waist-hip ratio on the prevalence of hypertension were significant, but the influence of waist circumference on hypertension was greater than waist-hip ratio. Keeping normal bodyweight might be one of the effective hypertension prevention measures.

[Studies on quality specification standards for Saposhnicovia divaricata (Turcz.) Schischk].

OBJECTIVE: To work out a qualitative-quantitative standard for Saposhnikovia divaricata. METHOD: TLC and HPLC were adopted. RESULT: Four chromone compounds were identified by TLC, and the contents of two chromone compounds were determined by HPLC. CONCLUSION: Prim-O-glucosylcimifugin, 4'-O-beta-D-glucosyl-5-O-methylvisamminol, sec-O-glucosylhamaudol and cimifugin should be present simultaneously in S. divaricata. The contents of prim-O-glucosylcimifugin and 4'-O-beta-D-glucosyl-5-O-methylvisamminol in S. divaricata should not be lower than 0.12% respectively.

[Studies on chemical constituents from root of Saposhnikovia divaricata (Turcz.) Schischk].

OBJECTIVE: To study the chemical constituents in the root of Saposhnikovia divaricata. METHOD: The compounds were isolated and identified by column chromatography and 1H, 13C NMR data. RESULT: Twenty-four compounds were isolated and identified. CONCLUSION: Seven compounds were isolated from this plant for the first time.

[Glucosides from Pleurospermum rivulorum].

AIM: To study the chemical constituents of Pleurospermum rivulorum. METHODS: To separate the constituents of P. rivulorum by using various kinds of chromatography and identify their structures on the basis of spectral analysis. RESULTS: Four glucosides were isolated from P. rivulorum. Their structures were established as marmesinin (I), tert-O-beta-D-glucopyranosyl-heraclenol (II), 1'-O-beta-D-glucopyranosyl (2R, 3S)-3-hydroxynodakenetin (III) and 8-hydroxy-5-O-beta-D-glucopyranosyl-psoralen (IV) based on the analysis of spectral data of IR, UV, MS, 1D and 2D-NMR. CONCLUSION: Compounds III and IV are new compounds. All of the four compounds were isolated from this plant for the first time.

Three novel cyclospirobifuranocoumarins, cyclorivulobirins A-C, from Pleurospermum rivulorum.

Three novel cyclospirobifuranocoumarins, cyclorivulobirins A-C (1-3), were isolated from the underground part of Pleurospermum rivulorum. They are characterized as three stereoisomers having a different configuration at the C-2 and C-2' position resulting from the condensation of two heraclenol units, respectively, on the basis of chemical and spectral methods.

Inhibitory effect of natural furanocoumarins on human microsomal cytochrome P450 3A activity.

To investigate the possible drug interaction with herbal medicine, furanocoumarin derivatives isolated from several Umbelliferous crude drugs were examined for their inhibitory effects on a typical human drug metabolizing enzyme, cytochrome P450 3A (CYP3A). Most furanocoumarins tested at 0.1 mM reduced microsomal testosterone 6beta-hydroxylation as an index of CYP3A activity to less than 50% of the control. In particular, the dimer and trimer derivatives of furanocoumarins showed striking inhibition, whose potencies were similar to that of a typical CYP3A inhibitor, ketoconazole. Preincubation of dimer types of furanocoumarins increased suppression but not most of the monomer derivatives, suggesting that the inhibition on CYP3A activity was caused by at least plural mechanisms. These results raised the possibility that the furanocoumarin containing herbal medicines may alter pharmacokinetics of co-ingested drugs similar to the case with grapefruit juice.

[A pharmacodynamic research on chromone glucosides of fangfeng].

OBJECTIVE: To investigate the pharmacological activity of prim-O-glucosylcimifugin and 4'-O-beta-D-glucosyl-5-O-methylvisamminol con tained in Fangfeng. METHODS: Analgesic, antipyretic and antiinflammatory effects were assessed using the models of yeast-induced rat fever, acetic acidinduced peritoneal pain thermalpain, and dimethylbenzol-induced mouse ear edema. RESULTS: Prim-O-glucosylcimifugin and 4'-O-beta-D-glucosyl-5-O-methylvisamminol could reduce the body temperature of rats in a model of yeast-induced fever. Both components could decrease the spasm action induced by peritoneal injection of acetic acid, increase the pain-threshold in a thermal-pain model, relieve the mouse ear edema caused by dimethybenzol and significantly inhibit ADP-induced platelet aggregation. CONCLUSION: Prim-O-glucosylcimifugin and 4'-O-beta-D-glucosyl-5-O-methylvisamminol have significant analgesic, antipyretic, anti-inflammatory and anti-platelet aggregation actions.

Involvement of p38 MAPK and ERK/MAPK pathways in staurosporine-induced production of macrophage inflammatory protein-2 in rat peritoneal neutrophils.

Stimulation of rat peritoneal neutrophils with staurosporine (64 nM) induced production of macrophage inflammatory protein-2 (MIP-2) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase/MAP kinase (ERK/MAPK). The staurosporine-induced MIP-2 production at 4 h was inhibited by the highly specific p38 MAPK inhibitor SB 203580 and the MAPK/ERK kinase (MEK-1) inhibitor PD 98059 in a concentration-dependent manner. By treatment with SB 203580 (1 microM) or PD 98059 (50 microM), the staurosporine-induced increase in the levels of mRNA for MIP-2 was only partially lowered, although the staurosporine-induced MIP-2 production was completely inhibited. Consistent with the inhibition by the protein synthesis inhibitor cycloheximide, SB 203580 and PD 98059 inhibited MIP-2 production at 4 h either when added simultaneously with staurosporine or 2 h after stimulation with staurosporine. In contrast, the DNA-dependent RNA polymerase inhibitor actinomycin D did not inhibit MIP-2 production at 4 h when it was added 2 h after staurosporine stimulation. Dot blot analysis demonstrated that treatment with SB 203580 or PD 98059 down-regulates the stability of MIP-2 mRNA. These results suggested that p38 MAPK and ERK/MAPK pathways are involved in translation of MIP-2 mRNA to protein and stabilization of MIP-2 mRNA.

Pharmacological analysis of protein kinases responsible for chemotaxis of rat peritoneal neutrophils.

Several types of kinase inhibitors were used to investigate the possible signaling pathways leading to the chemotaxis of rat peritoneal neutrophils toward macrophage inflammatory protein-2, cytokine-induced neutrophil chemoattractant-1, and platelet-activating factor. The chemotaxis and shape changes induced by each of these chemoattractants were strongly inhibited by a tyrosine kinase inhibitor (herbimycin A) and protein kinase C inhibitors (H-7 (1-(5-isoquinolinesulphonyl)-2-methylpiperazine dihydrochloride) and calphostin C). The formation of phosphatidyl 3,4,5-triphosphate in chemoattractant-stimulated neutrophils was completely inhibited by 100 nM of wortmannin, an inhibitor of phosphatidylinositol 3-kinase, whereas the chemotaxis toward each of these chemoattractants was partially inhibited (50% inhibition). The mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK-1) inhibitor PD 98059 did not inhibit the neutrophil chemotaxis. These findings suggest that the activation of tyrosine kinase and protein kinase C strongly participates in neutrophil chemotaxis and that the activation of phosphatidylinositol 3-kinase is partially involved, but that the activation of mitogen-activated protein kinase is not involved in neutrophil chemotaxis. The cross-linking of the signaling pathways for chemotaxis toward each chemoattractant was also examined.

Inhibition of 15-hydroxyprostaglandin dehydrogenase activity in rabbit gastric antral mucosa by panaxynol isolated from oriental medicines.

Panaxynol is a polyacetylene compound with anti-inflammatory and anti-platelet-aggregatory effects isolated from commonly used oriental medicines. The effects of panaxynol on the activity of prostaglandin-synthesizing and catabolizing enzymes in the rabbit gastric antral mucosa have been examined. At concentrations ranging from 25 to 200 microM panaxynol had no effect on the synthesis of prostaglandins E2, F2alpha and D2 from exogenous arachidonic acid in the microsomal fraction of the gastric mucosa whereas at 25-200 microM it dose-dependently inhibited the activity of 15-hydroxyprostaglandin dehydrogenase (PGDH), which catalyses the initial step of prostaglandin catabolism, in the cytosolic fraction. The concentration required for 50% inhibition (IC50) was approximately 25 microM. Inhibition of PGDH by panaxynol was non-competitive with regard to NAD+ and prostaglandin E2. These results suggest that panaxynol has the potential to inhibit PGDH activity in gastric mucosa, possibly as a result of pharmacological activity.

Induction of neutrophil chemotactic factor production by staurosporine in rat peritoneal neutrophils.

1. Incubation of rat peritoneal neutrophils in medium containing various concentrations of staurosporine (6.4-64 nM) increased the neutrophil chemotactic activity in the conditioned medium in a time- and concentration-dependent manner. 2. Separation of the neutrophil chemotactic activity in the conditioned medium by isoelectric focusing revealed that staurosporine (64 nM) stimulated the production of basic (pH > 8) neutrophil chemotactic factors, while TPA (12-O-tetradecanoylphorbol 13-acetate, 49 nM) stimulated the production of both basic (pH > 8) and acidic (pH 5) neutrophil chemotactic factors. 3. Determination by immunoassay of cytokine-induced neutrophil chemoattractant (CINC)-1, -2 alpha, -2 beta and -3 in the conditioned medium at 4 h revealed that staurosporine (64 nM) and TPA (49 nM) strongly stimulated the production of CINC-3 (staurosporine, 133.0 +/- 3.8; TPA, 26.7 +/- 1.0; control, 0.32 +/- 0.01 ng ml-1, means +/- s.e.mean from four samples) compared to CINC-1 (staurosporine, 55.0 +/- 1.2; TPA, 12.2 +/- 0.3; control, 0.56 +/- 0.01 ng ml-1), and CINC-2 alpha (staurosporine, 1.09 +/- 0.03; TPA, 0.90 +/- 0.02; control, < 0.10 ng ml-1). CINC-2 beta was below the detectable amount (< 0.078 ng ml-1). 4. The level of CINC-3 mRNA in the peritoneal neutrophils was determined by reverse transcription-polymerase chain reaction. Staurosporine (64 nM) and TPA (49 nM) enhanced the level of CINC-3 mRNA time-dependently, but had no effect on GAPDH mRNA levels. 5. Production of staurosporine-induced neutrophil chemotactic factor was inhibited by the protein kinase C inhibitors, H-7 (IC50, 12.3 microM), calphostin C (IC50, 0.77 microM) and Ro 31-8425 (24.3% inhibition at 10 microM), and by the tyrosine kinase inhibitor, genistein (IC50, 68.5 microM). Production of TPA-induced neutrophil chemotactic factor was also inhibited by both inhibitors. 6. Both the staurosporine- and the TPA-induced increase in CINC-3 mRNA levels were suppressed by H-7 and genistein.

Possible participation of macrophage inflammatory protein 2 in neutrophil infiltration in allergic inflammation in rats.

Recombinant rat macrophage inflammatory protein 2 (MIP-2) was prepared from E. coli transfected with a glutathione-S-transferase (GST)-MIP-2 fusion protein expression vector. A polyclonal antibody to rat MIP-2 was then obtained from rabbits by immunization with recombinant rat MIP-2. Using the polyclonal antibody which selectively suppressed neutrophil chemotactic activity of MIP-2, the role of MIP-2 in neutrophil infiltration in allergic inflammation in rats was studied. In an air pouch-type allergic inflammation model in rats, neutrophil infiltration into the pouch fluid increased with time after antigen challenge. Neutrophil chemotactic activity in the pouch fluid collected 8 h after antigen challenge was diminished by anti-MIP-2 antibody. In addition, when leukocytes that had infiltrated into the pouch fluid collected 4 h after antigen challenge were incubated, neutrophil chemotactic activity in the conditioned medium increased time-dependently, and the activity was neutralized by anti-MIP-2 antibody. Furthermore, when anti-MIP-2 antibody was injected into the pouch 6 h after antigen challenge, neutrophil infiltration into the pouch fluid during the next 2 h was suppressed. These findings indicate that MIP-2 plays an important role in neutrophil infiltration in rat allergic inflammation.