Adjustable rotation of multiple vortices produced by diode-pumped Nd:YVO4 lasers using intracavity second harmonic generation.

The criteria for achieving adjustable rotation of optical vortices are analyzed and used to design a diode-pumped solid-state laser that incorporates intracavity second harmonic generation within a concave-flat cavity to produce frequency-doubled Hermite-Gaussian (FDHG) modes. These FDHG modes are subsequently employed to generate various structured lights containing 2, 4, and 6 nested vortices using an external cylindrical mode converter. Through theoretical exploration, we propose that increasing the radius of curvature of the concave mirror and extending the cavity length can enhance the rotational angles of multiple vortices by expanding the adjustable range of phase shift for FDHG modes. Moreover, theoretical analyses assess vortex rotation concerning the positions of a nonlinear medium, successfully validating the experimental observations and elucidating the phase structures of the transformed beams.

Impact of Obesity on Timing of Tracheotomy: A Multi-institutional Retrospective Study.

OBJECTIVE: To examine the impact of increased body mass index (BMI) on (1) tracheotomy timing and (2) short-term surgical complications requiring a return to the operating room and 30-day mortality utilizing data from the Multi-Institutional Study on Tracheotomy (MIST). METHODS: A retrospective analysis of patients from the MIST database who underwent surgical or percutaneous tracheotomy between 2013 and 2016 at eight institutions was completed. Unadjusted and adjusted logistic regression analyses were used to assess the impact of obesity on tracheotomy timing and complications. RESULTS: Among the 3369 patients who underwent tracheotomy, 41.0% were obese and 21.6% were morbidly obese. BMI was associated with higher rates of prolonged intubation prior to tracheotomy accounting for comorbidities, indication for tracheotomy, institution, and type of tracheostomy (p = 0.001). Morbidly obese patients (BMI ≥35 kg/m2) experienced a longer duration of intubation compared with patients with a normal BMI (median days intubated [IQR 25%-75%]: 11.0 days [7-17 days] versus 9.0 days [5-14 days]; p < 0.001) but did not have statistically higher rates of return to the operating room within 30 days (p = 0.12) or mortality (p = 0.90) on multivariable analysis. This same finding of prolonged intubation was not seen in overweight, nonobese patients when compared with normal BMI patients (median days intubated [IQR 25%-75%]: 10.0 days [6-15 days] versus 10.0 days [6-15 days]; p = 0.36). CONCLUSION: BMI was associated with increased duration of intubation prior to tracheotomy. Although morbidly obese patients had a longer duration of intubation, there were no differences in return to the operating room or mortality within 30 days. LEVEL OF EVIDENCE: III Laryngoscope, 2024.

Highly selective microbial transformation of major ginsenoside Rb1 to gypenoside LXXV by Esteya vermicola CNU120806.

AIMS: This study examined the biotransformation pathway of ginsenoside Rb(1) by the fungus Esteya vermicola CNU 120806. METHODS AND RESULTS: Ginsenosides Rb(1) and Rd were extracted from the root of Panax ginseng. Liquid fermentation and purified enzyme hydrolysis were employed to investigate the biotransformation of ginsenoside Rb(1) . The metabolites were identified and confirmed using NMR analysis as gypenoside XVII and gypenoside LXXV. A mole yield of 95·4% gypenoside LXXV was obtained by enzymatic conversion (pH 5·0, temperature 50°C). Ginsenoside Rd was used to verify the transformation pathway under the same reaction condition. The product Compound K (mole yield 49·6%) proved a consecutive hydrolyses occurred at the C-3 position of ginsenoside Rb(1) . CONCLUSIONS: Strain CNU 120806 showed a high degree of specific ß-glucosidase activity to convert ginsenosides Rb(1) and Rd to gypenoside LXXV and Compound K, respectively. The maximal activity of the purified glucosidase for ginsenosides transformation occurred at 50°C and pH 5·0. Compared with its activity against pNPG (100%), the ß-glucosidase exhibited quite lower level of activity against other aryl-glycosides. Enzymatic hydrolysate, gypenoside LXXV and Compound K were produced by consecutive hydrolyses of the terminal and inner glucopyranosyl moieties at the C-3 carbon of ginsenoside Rb(1) and Rd, giving the pathway: ginsenoside Rb(1) → gypenoside XVII → gypenoside LXXV; ginsenoside Rd→F(2) →Compound K, but did not hydrolyse the 20-C, ß-(1-6)-glucoside of ginsenoside Rb(1) and Rd. SIGNIFICANCE AND IMPACT OF THE STUDY: The results showed an important practical application on the preparation of gypenoside LXXV. Additionally, this study for the first time provided a high efficient preparation method for gypenoside LXXV without further conversion, which also gives rise to a potential commercial enzyme application.

Effect of Wnt signaling pathway on wound healing.

Wnt signaling pathway has been divided into two subclasses: the canonical pathway (Wnt/beta-catenin pathway) and the non-canonical pathway. It has been proven that Wnt/beta-catenin pathway can enhance wound healing, and some glycoprotein of Wnt family may directly or indirectly improve wound healing.

Effects of lithium and insulin on glycogen synthesis in L6 myocytes: additive effects on inactivation of glycogen synthase kinase-3.

In insulin-sensitive L6 myocytes, insulin stimulated glycogen synthesis in a dose-dependent manner and lithium further stimulated glycogen synthesis at all insulin concentrations. Lithium alone at 20 mM stimulated glycogen synthesis to the degree similar to the maximal insulin response. Effects of lithium and insulin were fully additive for both glycogen synthesis and glycogen synthase activity. In L6 myocytes, insulin increased phosphorylation of Akt1 and glycogen synthase kinase-3 alpha and beta (GSK-3 alpha and beta), resulting in its activation and inactivation, respectively. Unlike insulin, lithium directly inhibited GSK-3 (both alpha and beta) without affecting phosphorylation of GSK-3. Moreover, lithium in vitro could further inhibit enzyme activity of GSK-3 (both alpha and beta) that was isolated from insulin-stimulated cells (thus already phosphorylated and inactivated by insulin). In summary, insulin increases glycogen synthesis by the Akt1/GSK-3/glycogen synthase pathway, but lithium increases glycogen synthesis by direct inhibition of GSK-3 in L6 myocytes. Inhibitory effects of lithium and insulin on GSK-3 (both alpha and beta) were additive, which may account, at least in part, for their additive effects on glycogen synthase activity and glycogen synthesis in L6 myocytes.

Regulation of protein phosphorylation in pancreatic acini by cyclic AMP-mediated secretagogues: interaction with carbamylcholine.

The effects on protein phosphorylation in mouse pancreatic acini of cyclic AMP-mediated secretagogues and the Ca2+-mediated agonist carbamylcholine were compared. Under the conditions adopted for the study of protein phosphorylation, carbamylcholine (3 microM) stimulated amylase release from pancreatic acini 6-fold, whereas vasoactive intestinal polypeptide (VIP) (100 nM) and the cyclic AMP analogue 8-bromo-cyclic AMP (1 mM) caused little or no increase in secretion. However, VIP and 8-bromo-cyclic AMP, when added in combination with carbamylcholine, potentiated the stimulation of amylase release to 170-180% of that caused by carbamylcholine alone. As assessed by two-dimensional gel electrophoresis, VIP reproduced four of the ten changes in protein phosphorylation elicited by carbamylcholine, these changes being the increased phosphorylation of one soluble protein and the decreased phosphorylation of three soluble proteins. VIP enhanced the carbamylcholine-induced changes in phosphorylation for three proteins. In addition, VIP increased the phosphorylation of a unique protein of Mr 52,000 and pI 5.66 which was not affected by carbamylcholine. All of the effects on protein phosphorylation exerted by VIP in the presence or absence of carbamylcholine were mimicked by 8-bromo-cyclic AMP. Secretin also reproduced most of the changes in protein phosphorylation caused by VIP, although concentrations of secretin of at least 100-fold higher were required to elicit a maximal response. It is concluded that cyclic AMP-mediated secretagogues alter the phosphorylation of a unique protein as well as of several pancreatic proteins affected by carbamylcholine. Moreover, these effects appear to be mediated primarily by VIP-preferring receptors and may be involved in the synergistic action of VIP to promote carbamylcholine-induced amylase release.

Insulin and ribosomal protein S6 kinase in rat pancreatic acini.

Treatment of pancreatic acini from diabetic rats with insulin resulted in a dose-dependent increase in the phosphorylation of ribosomal protein S6 when analyzed by two-dimensional gel electrophoresis. To study the presence of the protein kinase mediating this phosphorylation, soluble extracts of intact acini that had been previously treated with insulin were prepared and assayed for protein kinase activity with rat pancreatic ribosomes as a substrate. Activation of S6 kinase activity, observed in a time-dependent manner, was maximal after 20-30 min and, in a dose-dependent manner, was half-maximal at 1 nM and maximal at 10 nM insulin concentration. Based on cofactor requirements, substrate specificity, and a slow activation of the enzyme, the S6 kinase was distinct from cAMP-dependent, Ca2+-calmodulin-dependent, and Ca2+-phospholipid-dependent protein kinases and protease-activated kinase II. The S6 kinase activated by insulin was highly specific for the ribosomal protein S6 when compared with various substrates, including casein, glycogen synthase, phosphorylase b, phosvitin, histone HIII-S, and histone HVIII-S. Protein S6 phosphorylation in intact acini and activation of the S6 kinase by insulin showed similar dose-response curves, consistent with the S6 kinase being responsible for the protein S6 phosphorylation in intact acini. The comparison of the dose-response curves for S6 phosphorylation and protein synthesis in acini suggests that there is a close correlation between these two insulin actions.

Deletion of residues 485-599 from the human insulin receptor abolishes antireceptor antibody binding and influences tyrosine kinase activation.

We have studied insulin and antireceptor antibody binding to mutated human insulin receptors deleted of residues 485-599 in the alpha-subunit by site-directed mutagenesis. Both normal and mutated receptors were expressed in rat HTC hepatoma cells. Cells expressing either the normal receptor or the mutated receptor retained the ability to bind insulin. In contrast to the normal receptor, however, the mutated receptor failed to interact with antireceptor alpha-subunit antibodies. The inability of the mutated receptor to interact with various antireceptor antibodies was further documented by photoaffinity labeling studies. In intact HTC cells expressing mutated receptors, basal insulin receptor tyrosine autophosphorylation was 2-fold elevated when compared to cells expressing normal receptors. In these cells, however, the response of this function to insulin was blunted. When receptors were isolated from these cells and assayed for both autophosphorylation and phosphotransferase activities toward the synthetic substrate poly(Glu, Tyr), the response to insulin was also blunted. To study the ability of the mutated receptor to transmembrane signal, insulin stimulation of S6 kinase activity was measured. In cells with mutated receptors, in concert with the insulin receptor kinase data, basal S6 kinase activity was elevated, and the response to insulin was blunted. The data suggest, therefore, that residues 485-599 in the alpha-subunit of the insulin receptor are critical for antireceptor antibody binding, but not for insulin binding. Moreover, these data suggest that residues 485-599 contain a regulatory domain for insulin regulation of receptor beta-subunit functions.

Role of p85 subunit of phosphatidylinositol-3-kinase as an adaptor molecule linking the insulin receptor to insulin receptor substrate 1.

After insulin stimulation of cells, signaling complexes are formed, containing the insulin receptor (IR), insulin receptor substrate-1 (IRS-1), and phosphatidylinositol-3-kinase. To study the nature of these complexes, we employed purified IR, recombinant IRS-1, antibodies to IR and IRS-1, and fusion proteins containing the two SH2 domains of p85. In intact cells, insulin increased tyrosine phosphorylation of both the IR and IRS-1. Both of these proteins were immunoprecipitated with antibodies to p85. Also, fusion proteins containing the two SH2 domains of p85 directly precipitated both the IR and IRS-1. Next, these signaling complexes were reconstituted in vitro with purified IR, recombinant IRS-1, and the two SH2 domains of p85. In the presence of both SH2 domains of p85, the IR associated with IRS-1. Other data, both in intact cells and in vitro, demonstrated that N- and C-terminal SH2 domains of p85 had preferential binding affinities for the IR and IRS-1, respectively. Studies with an IR mutant truncated in the C terminus indicated that the C-terminal phosphotyrosines of the IR play a major role in interacting with the SH2 domains of p85. In conclusion, both in vivo and in vitro data support a role for p85 in directly linking the IR to IRS-1 via its SH2 domains. The formation of these complexes, therefore, may provide a mechanism for the translocation to the plasma membrane of phosphatidylinositol-3-kinase and other molecules that are involved in IR signaling.

Natural medicines for alcoholism treatment: a review.

Alcoholism is a serious problem throughout the world. The development of alcoholism remedies have medical, social and economical significance. In view of the pitfalls of psychological dependence and adverse behavioural effects of synthetic drugs, the development of low toxicity and high efficiency medicines derived from natural products exhibits expansive market prospects. Based on these considerations, we summarize briefly folk application of traditional hangover remedies and clinical application of herbal complex and patent medicines for alcoholism treatment. We have reviewed the effects of natural medicines on intake, absorption and metabolism of alcohol, as well as the protective effects on alcohol-induced acute and chronic tissue injury.

Insulin-like growth factor-1 stimulation of cells induces formation of complexes containing phosphatidylinositol-3-kinase, guanosine triphosphatase-activating protein (GAP), and p62 GAP-associated protein.

The insulin-like growth factor-1 (IGF-1) receptor is structurally related to the insulin receptor and shares common features in receptor signaling. These features include receptor autophosphorylation, phosphorylation of insulin receptor substrate-1, and activation of Ras and phosphatidylinositol-3-kinase (PI3K). Previously, we reported that after insulin treatment of rat HTC cells expressing human insulin receptors, a unique insulin receptor signaling complex was formed that contained the insulin receptor, the p85 subunit of PI3K, GTPase-activating protein (GAP), and p62 GAP-associated protein. In the present study, using wild type HTC cells, we investigated whether the activated IGF-1 receptor also forms a similar signaling complex. To study the proteins present in IGF-1 receptor signaling complexes, we used immunoprecipitation and Western blotting analysis with appropriate antibodies. In response to IGF-1, insulin receptor substrate-1 was tyrosine phosphorylated and formed a complex with the PI3K heterodimer that consists of a p85 regulatory subunit and a p110 catalytic subunit. In addition, a separate complex was formed, consisting of p85, p62 GAP-associated protein and GAP. The p62 in this complex was tyrosine phosphorylated. These studies suggest, therefore, that the IGF-1 receptor, like the insulin receptor, induces the formation of multiple signaling complexes that most likely mediate the proliferative effects of these receptors.

Guanosine triphosphatase-activating protein-associated protein, but not src-associated protein p68 in mitosis, is a part of insulin signaling complexes.

The insulin receptor, following insulin stimulation of cells, triggers formation of various signaling complexes. In rat HTC hepatoma cells overexpressing normal human insulin receptors (HTC-IR), p85 regulatory subunit of phosphatidylinositol-3-kinase (PI3K) forms signaling complexes containing the insulin receptor, insulin receptor substrate 1 (IRS-1), guanosine triphosphatase-activating protein (GAP) and 60-70 kDa phosphotyrosine proteins (p60-70). In the present study, we demonstrate that p60-70 interacts directly with the p85 subunit via src homology 2 domain of the latter. Employing antibodies specific to two p85 isoforms, p85alpha and p85beta, we demonstrate that HTC-IR cells express both p85 isoforms, and these isoforms induce the formation of similar signaling complexes in response to insulin. p60-70, present in both alpha-p85alpha and alpha-p85beta immunoprecipitates, is a GAP-associated protein, but is distinct from the p68 src-associated protein in mitosis (Sam68) by several criteria. These data suggest that 1) GAP-associated protein, but not Sam68, is a part of insulin signaling complexes; and 2) p85alpha and p85beta form similar, but distinct, insulin receptor signaling complexes.

Assessment of viability of Schistosoma mansoni miracidia by Congo red staining.

The viability of Schistosoma mansoni miracidia, either hatched or unhatched, has been assessed by a staining technique using Congo red, a pH-sensitive dye (pH 3-5), under the light microscope. Hatched, free-swimming miracidia and live miracidia inside egg shells were stained light blue whereas dying miracidia altered the dye color from blue to orange and eventually to red. Immature eggs containing undeveloped miracidia were not stained by Congo red. This staining technique is simple, rapid and can be used as a method to assess both the viability of S. mansoni miracidia and the hatching process.

Cholecystokinin stimulates a specific ribosomal S6 kinase in rat pancreatic acini.

Stimulation of intact rat pancreatic acini with cholecystokinin (CCK) enhances the phosphorylation of the ribosomal protein S6 in a dose-dependent manner with half maximal stimulation at 40 pM and maximal stimulation at 1 nM CCK octapeptide. Soluble cellular extracts contained S6 kinase activity assayed using purified rat pancreatic ribosomes as substrate. Stimulation by CCK of S6 kinase was concentration dependent, being half maximal at 50 pM and maximal at 1 nM CCK. The phorbol ester 12-O-tetradecanoyl-phorbol 13-acetate (TPA), an activator of protein kinase C, also increased both S6 phosphorylation in intact acini and soluble S6 kinase activity. In order to determine whether S6 kinase mediated S6 phosphorylation following CCK treatment of acini, two-dimensional phosphopeptide analysis was performed for S6 proteins phosphorylated under various conditions. These data suggest that a specific soluble S6 kinase, the activation of which appears to be directly or indirectly mediated by protein kinase C, is the functional enzyme in intact acini that mediates the action of CCK to increase S6 phosphorylation and may be involved in increased protein synthesis in pancreatic acini treated with CCK.

Percutaneous gallbladder drainage for delayed laparoscopic cholecystectomy in patients with acute cholecystitis.

BACKGROUND: Many studies have concluded that delayed or interval laparoscopic cholecystectomy (LC) in patients with acute cholecystitis (AC) demonstrated higher conversion rates and complication rates compared with early LC. However, if the acutely inflamed gallbladder is decompressed by emergent percutaneous gallbladder drainage (PGBD), it may decrease the technical difficulty of LC allowing successful delayed LC when the patient is in better condition. The purpose of this retrospective study was to assess the outcomes of delayed LC following PGBD in patients with AC. METHODS: A total of 72 LC for AC were divided into PGBD (n = 27) and non-PGBD groups (n = 45). The PGBD group had delayed LC (after 72 hours of admission). Thirty-two non-PGBD patients had early LC (within 72 hours of admission) and 13 non-PGBD had delayed LC. Outcome of delayed LC for the PGBD group was assessed by LC time, conversion rate, morbidity rate, and hospital stay, and compared with that of the non-PGBD group. RESULTS: Compared with early and delayed LC of the non-PGBD group, the PGBD group showed longer LC time (median 110 minutes versus 87.5 minutes versus 85 minutes, P <0. 05), a little lower conversion rate (15% versus 25% versus 23%), similar morbidity rate (15% versus 9% versus 15%), and prolonged hospital stay (13 days versus 7 days versus 10 days). CONCLUSIONS: PGBD did not significantly improve the outcome of LC for AC as assessed by conversion and morbidity rate and hospital stay compared with no PGBD. Thus, we can conclude that although PGBD is a safe and effective emergency procedure for AC, it should be limited to higher risk groups such as elderly or critically ill patients and to acalculous cholecystitis.

Cysteinyl proteinases of Schistosoma mansoni eggs: purification and partial characterization.

The egg stage of Schistosoma mansoni, a trematode blood fluke, is known to be responsible for an immunologically mediated granuloma formation. Proteolytic enzymes of S. mansoni eggs may be involved in the penetration of host tissue by eggs and/or may act as antigens to cause a humoral as well as a cell-mediated response leading to granuloma formation. Three acidic, thiol-dependent proteinases from the eggs of S. mansoni were isolated, and 2 major proteinases (I and II) were purified to homogeneity using chromatofocusing, AcA54 ultrogel chromatography, and thiopropyl-Sepharose 6B affinity chromatography. Proteinases I and II have molecular weights of 25,400 and 30,500, and isoelectric points of 6.0 and 5.6, respectively. These enzymes were found to be cathespin B-like cysteinyl proteinases based on similarities in molecular weight, isoelectric point, optimal assay pH, instability to neutral pH, substrate specificity, and inhibitor sensitivity. A monoclonal antibody, specific to S. mansoni egg proteinases was used in immunoblotting studies. Under native, but not under denaturing, conditions for gel electrophoresis, this monoclonal antibody reacted with egg proteinases. This antibody had previously been shown to recognize an antigen in the miracidial penetration glands of schistosome eggs.

Prediction of common bile duct stones: its validation in laparoscopic cholecystectomy.

BACKGROUND/AIMS: Although perioperative cholangiography is valuable and highly accurate in the detection of common bile duct (CBD) stones, its routine use is controversial, particularly in the era of the laparoscopic cholecystectomy because of its inherent disadvantages. The purposes of this retrospective and prospective study on cholelithiasis were to identify patients at low risk for CBD stones and to assess the validity of the low risk criteria. METHODOLOGY: For the first, retrospective study, 15 significant preoperative clinical, biochemical and sonographic variables were selected from 561 consecutive patients who underwent conventional cholecystectomy with routine intraoperative cholangiography (IOC) for cholelithiasis from January 1985 to December 1993, and independent risk factors predicting the presence of CBD stones were determined by multivariate logistic regression analysis. For the second, prospective study, from April 1994 to September 1995, a laparoscopic cholecystectomy (LC) was performed without perioperative cholangiography in 153 consecutive patients with the primary low risk criterion (sonographic CBD diameter < 10 mm) determined by the first study. All of the LC patients were followed-up for a median duration of 12 months (range 4 to 21 months). RESULTS: In the first study, CBD stones were present in 95 (16.9%) patients. The most important independent predictor was a dilated CBD (> 10 mm). Three levels of risk were determined: (1) the low risk group (73.8% of the patients), in which the CBD was not dilated and the prevalence of CBD stones was 1.5% (6/408); the moderate risk group (7.8% of the patients), in which there was a dilated CBD with normal liver function tests and a prevalence of stones of 48.8% (21/43); and the high risk group (18.4%), in which there was a dilated CBD and abnormal liver function tests and a prevalence of stones of 66.7% (68/102). In the second study, two cases (1.4%) of symptomatic overlooked CBD stones were found on endoscopic retrograde cholangiography and retrieved by endoscopic sphincterotomy on postoperative days 18 and 20, respectively. CONCLUSIONS: Preoperative assessment in cases of cholelithiasis can determine which patients are at low risk for having CBD stones, thereby avoiding unnecessary perioperative cholangiography. This selectivity is also valid in LC, since the incidence of symptomatic, overlooked CBD stones was very low.

Production of antibody against saikosaponin a, an active component of bupleuri radix.

High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, inoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested.

Metabolism of saikosaponin c and naringin by human intestinal bacteria.

By human intestinal bacteria, saikosaponin c was transformed to four metabolites, prosaikogenin E1 (E1) prosaikogenin E2 (E2), prosaikogenin E3 (E3) and saikogenin E. Metabolic time course of saikosaponin c was as follows; in early time, saikosaponin c was converted to E1 and E2, and then these were transformed to saikogenin E via E3. Also, this metabolic pathway was similar to the metabolism of saikosaponin c by rat intestinal bacteria.Bacteroides JY-6 andBacteroides YK-4, the bacteria isolated from human intestinal bacteria, could transform saiko-saponin c to E via E1 (or E2) and E3. However, these bacteria were not able to directly transform E1 and E2 to saikogenin E. Naringin was mainly transformed to naringenin by human intestinal bacteria. The minor metabolic pathway transformed naringin to naringenin via prunin. By JY-6 or YK-4, naringin was metabolized to naringenin only via prunin.

In vitro angiogenic activity of Aloe vera gel on calf pulmonary artery endothelial (CPAE) cells.

Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type 1 collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the mRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitor-1 (PAI-1) mRNA was not changed.