Automated preparation of washed platelet concentrates through spinning-membrane filtration.

BACKGROUND: Washed platelet concentrates (WPC), prepared with an automated system cell processor (ACP), have recently been approved to be manufactured and marketed in Japan. From the perspective of risk management, it is preferable to secure alternative technologies for ACP. Here, we conducted a study to evaluate the quality of WPC prepared using an automated membrane filtration-based system, Lovo. STUDY DESIGN AND METHODS: Replaced PCs prepared from apheresis PCs were equally divided into control and test units, and subsequently washed using ACP and Lovo respectively. Work and operational efficiencies were evaluated by in vitro analyses, including total handling time, platelet recovery, and plasma protein removal rate. Product quality, including a set of biochemical and physiological indicators of platelets and supernatants, were assessed before and 3 days after washing. RESULTS: In vitro platelet recovery rates and plasma protein removal rates were >85% and >95%, respectively, in both groups. The pH values on day 0 were significantly high (6.97 vs. 6.86) due to low pCO2 in the test group, while no significant differences in glucose consumption and lactate production were observed between the two groups. The levels of hypotonic shock responses, aggregation response, platelet shape, CD62P expression, and sCD62P concentration were similar in both groups during the 3-day storage period. CONCLUSION: Platelet washing with Lovo provides platelet quality equivalent to, or better than, conventional washing with ACP. Thus, the new automated system, Lovo, can be considered as an alternative to ACP for WPC preparation.

Prevalence and Trends in Alcohol Dependence and Alcohol Use Disorders in Japanese Adults; Results from Periodical Nationwide Surveys.

AIMS: Nationwide surveys to clarify the characteristics and trends of the drinking behavior of Japanese adults were carried out in 2003, 2008, and 2013. METHODS: These were periodical cross-sectional surveys. Subjects were chosen through a stratified two-stage random sampling method. The surveys included drinking frequency and amount, ICD-10 alcoholism diagnostic standards, questionnaire for the determination of harmful alcohol use ( AUDIT: Alcohol Use Disorders Identification Test). In 2003, the surveys obtained responses from 2547 people (73% response rate); in 2008, 4123 people (55% response rate); and in 2013, 4153 people (59% response rate). RESULTS: The proportion of lifetime experience of alcohol dependence diagnosed by ICD-10 was 1.9% for male and 0.2% for female, and the estimated number of patients was 1.07 million. The declining trends were observed in the percentage of daily drinkers and the amount of alcohol consumed per week for male. The lowering of the age for consuming their first alcoholic drink and their first drunken experience was observed among female. The gender difference of prevalence of problem drinking is getting smaller. The binge drinking and heavy episodic drinking were observed especially younger generation. The only small proportion of patients with alcohol dependence had received specialized medical care, whereas the many of these visited medical institutions and health screening. CONCLUSIONS: The survey observed many hidden alcoholic patients, and showed the possibility that the healthcare facilities and health screening became the place of screening and intervention for alcohol dependence.

Current Status of Alcoholic Liver Disease in Japan and Therapeutic Strategy.

Alcoholism results in an estimated 3.3 million deaths annually worldwide, accounting for 5.9% of all mortality. Although per capita alcohol consumption in Japan, had been gradually decreasing since 1999, it has plateaued in the past 5 years. Alcoholic liver disease (ALD) is the most prevalent cause of advanced liver disease, and includes alcoholic fatty liver, alcoholic hepatitis (AH), alcoholic fibrosis, alcoholic liver cirrhosis (ALC), and alcoholic hepatocellular carcinoma (HCQ. Although alcohol consumption is the predominant etiological factor in the pathogenesis of ALD, there is marked variation in liver mortality rates among. different countries and over time within countries. Six national surveys of ALD in Japan were carried out by the Japanese ALD study groups. The first three studies reported that the prevalence of ALD increased in parallel with an increase in alcoholic beverage -intake and that the rise in ALD was one major- factor contributing to the increased prevalence of liver cirrhosis. However; recent epidemiological studies- showed that ALC continues to rise despite a gradual decrease in alcohol intake, indicating that there are other risk factors for the development of ALC. Our recent survey revealed that the prevalence of ALC has been rapidly increasing in Japan and that the prevalence of alcoholic HCC in ALC was higher in elderly male patients and younger patients with diabetes mellitus (DM). DM, female sex, and age were identified as risk factors for the development of ALC, while DM, male sex, and age were identified as significant risk factors for HCC in ALC. Severe alcoholic hepatitis (SAH) is an inflammatory response with multiple morbidity factors like leucocytosis, hepatomegay, and renal failure, and has a high mortality rate. We have created a new scoring system for AH (Japan Alcoholic Hepatitis Score [JAS]). Its ability to predict outcome was confirmed by examining the data of 59 patients with AH in 2011: 26 had moderate AH of whom 22 were alive and were dead, while 33 had SAH of whom 16 were alive and 17 were dead. There was no report of death in patients with mild AH (JAS was ≤ 7). The prevalence of renal failure, DIC and gastrointestinal bleeding was higher in patients who had died, while Cr and PT (INR) were higher in SAH patients who had died. These results suggest that JAS allows stratification of the risk of death and can help manage patients with AH. Our belief is that patients with elevated myeloid leucocytes benefit most from granulocytes/monocytes apheresis, while exchange appears to support patients with coagulation deficiency of high plasma bilirubin and hemodialysis is indicated for hit Cr. Liver transplantation is the ultimate therapy for ALD, but it has always been controversial. In Japan, most cases are living donor liver transplantations because the number of brain-dead donors are low. Although 6 months of abstinence is required for brain-dead donor liver transplantation in most countries, 18 months of proven abstinence is required in Japan.

No contribution of the ABCB11 p.444A polymorphism in Japanese patients with drug-induced cholestasis.

European studies have revealed that the ABCB11 c.1331T>C (V444A) polymorphism (rs2287622) C-allele frequency is higher among patients with drug-induced cholestasis. Given the low incidence of this disease, however, this association has not been sufficiently elucidated. We aimed to investigate the significance of this polymorphism in Japanese patients. We determined ABCB11 V444A polymorphism frequencies and HLA genotypes in two independent drug-induced cholestasis cohorts. Expression and taurocholate transport activity of proteins from 444A variants were analyzed using Madin-Darby canine kidney II cells. In cohort 1 (n = 40), the V444A polymorphism C-allele frequency (66%) was lower than that in controls (n = 190, 78%), but this difference was not significant (P = 0.09). In cohort 2 (n = 119), comprising patients with cholestatic (n = 19), hepatocellular (n = 74), and mixed (n = 26) liver injuries, the C-allele frequency was lower among patients with cholestatic liver injury (68%) than among those with hepatocellular (75%) or mixed liver injury (83%), although this difference was not significant. In cohort 1, HLA-A*0201 was observed more frequently in patients (22%) than in controls [11%; P = 0.003; odds ratio, 2.4 (95% confidence interval, 1.4-4.0)]. Taurocholate transport activity of 444A-encoded protein was significantly lower than that of 444V-encoded protein (81% of 444V, P < 0.05) because of the reduced protein stability. In conclusion, ABCB11 444A had slightly reduced transport activity, but it did not contribute to the occurrence of drug-induced cholestasis in Japanese patients. Therefore, genetic susceptibility to acquired cholestasis may differ considerably by ethnicity.

Hepatocellular carcinoma based on cryptogenic liver disease: The most common non-viral hepatocellular carcinoma in patients aged over 80 years.

AIM: To clarify the clinical features of patients with hepatocellular carcinoma (HCC) with cryptogenic liver diseases, we analyzed the data from a nationwide survey in Japan. METHODS: The survey was conducted in 2009. The factors examined included age and underlying liver diseases: alcoholic liver disease (ALD; n = 991), non-alcoholic fatty liver disease (n = 292), modest alcohol intake (intake between 20 and 70 g/day, n = 214) and cryptogenic liver diseases (n = 316). We compared the clinical features of cryptogenic HCC among patient-age subgroups. RESULTS: HCC with ALD etiology was most common among the non-viral HCC patients under 80 years old; for those aged 80 years or older, cryptogenic HCC was the most common etiology. Among the cryptogenic HCC patients, the body mass index values and the prevalences of liver cirrhosis (LC) and diabetes mellitus (DM) were significantly lower in the 80 years or older group versus the 50-79 years group. In the 80 years or older group, 28% of the patients developed HCC without cirrhosis, obesity and DM. CONCLUSION: In the HCC patients aged 80 years and over, the etiology of most of the non-viral HCC cases was classified as cryptogenic. In light of our finding that the prevalences of obesity, DM and LC in the 80 years or older group of cryptogenic HCC patients were significantly lower those in the younger patients, it is apparent that analyses of HCC cases must take age differences into account.

Alcoholic Liver Cirrhosis and Significant Risk Factors for the Development of Alcohol-related Hepatocellular Carcinoma--Japan, 2012.

Recent epidemiological studies showed that alcoholic liver cirrhosis (ALC) and alcohol-related hepatocellular carcinoma (HCC) continue to increase despite a gradual decrease in alcohol intake, indicating that there are other risk factors for the development of ALC and alcohol-related HCC. A nationwide survey of liver cirrhosis (LC) was undertaken by asking major hospitals in Japan to provide information on the number of patients with LC admitted in 2012 along with information related to etiology including alcohol history and other relevant demographics. Among the 1,478 ALC patients, total alcohol intake was lower in women than in men, and mean age was also lower in women. Mean age was older in the present study in 2012 compared with the data in 2008. Compared to patients without diabetes mellitus (DM), patients with DM had lower daily alcohol intake and higher mean age. The prevalence of HCC was much higher in male elderly ALC patients and also in DM patients among the younger ALC patients (65 years). DM, female sex, and age were identified as risk factors for development of ALC, and DM, male sex, and age were identified as significant risk factors for HCC in ALC.

[Diagnostic accuracy for alcoholic liver disease with controlled Attenuation Parameter (CAP) measured by transient elastography for the non-invasive assessment of liver steatosis].

Along with the development of interferon and therapeutic medication, the incidence of viral hepatitis constituting the largest part of liver disease decreased, and the main target in the field of liver disease is now shifting from viral hepatitis to alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD) as metabolic liver disease. Although these diseases tend.to. be gathered as non-viral liver disease because the similar specific liver tissue, the natural history and etiology are considerably different between them. We need to distinguish both of them to do appropriate treatment intervention. Questioning of amount of drinking is needed, but we experience some difficult cases to understand drinking history because of a too little declaration of amount of drinking. A new ultrasonic image analyses using propagation speed in the organization of the pulse vibration wave was developed as Fibroscan by Echosens company in recent years. Fibroscan is a non-invasive test to quantify liver fibrosis as Liver Stiffness Measurement (LSM). It also detects and quantifies steatosis simultaneously using the Controlled Attenuation Parameter (CAP). CAP is a measurement of the ultrasound attenuation. We measured liver steatosis of patients using Fibroscan, and other blood tests. 63 cases of ALD, 177 cases of NAFLD, 57 cases of Virus and 271 cases of Normal were enrolled. CAP value were significantly lower in the ALD group compared with NAFLD group. (P < 0.0053, ALD 268 dB/m : NAFLD 290 dB/m) We elucidate the diagnostic accuracy of CAP using Fibroscan for ALD patients, comparing the results of them to those of virus patients and NAFLD patients.

Prevalence of hazardous drinking and suspected alcohol dependence in Japanese primary care settings.

OBJECTIVE: We aimed to assess the prevalence of hazardous drinking and potential alcohol dependence among Japanese primary care patients, and their readiness to change and awareness of others' concerns. METHODS: From July to August 2023, we conducted a multi-site cross-sectional study as a screening survey for participants in a cluster randomized controlled trial. The trial included outpatients aged 20-74 from primary care clinics. Using the Alcohol Use Disorders Identification Test (AUDIT) alongside a self-administered questionnaire, we evaluated the prevalence of hazardous drinking and suspected alcohol dependence, patients' readiness to change, and their awareness of others' concerns. RESULTS: Among the 1388 participants from 18 clinics, 22% (95% confidence interval (CI): 20% to 24%) were identified as engaging in hazardous drinking or suspected of being alcohol dependent. As the AUDIT scores increased, so did their readiness to change. However, only 22% (95%CI: 16% to 28%) of those with scores ranging from 8 to 14 reported that others, including physicians, had expressed concerns about their drinking during the past year. For those with scores of 15 or higher, the figure was 74%. CONCLUSIONS: This study underscores the need for universal or high-risk alcohol screening and brief intervention in Japanese primary care settings. Trial registry UMIN-CTR (https://www.umin.ac.jp/ctr/) (UMIN000051388).

Granulocytapheresis and plasma exchange for severe alcoholic hepatitis.

BACKGROUND AND AIM: Severe alcoholic hepatitis (SAH) is an inflammatory response with multiple morbidity factors like leucocytosis, hepatomegaly, renal failure, hepatic encephalopathy, endotoxemia, and a high mortality rate. Identifying therapeutic interventions that can improve prognosis is the goal of research. METHODS: Questionnaires were sent to 1234 medical institutions asking for information on patients with SAH during 2004 to 2008, including patients' demography, disease profile and the therapeutic interventions patients had received during hospitalization. RESULTS: Forty-five hospitals had treated SAH patients, and provided full demographic data on 98 patients. Forty-eight patients had received no treatment, 12 patients had received granulocytes/monocytes apheresis (GMA) to deplete elevated myeloid lineage leucocytes, the rest had received one or more of the following treatments, corticosteroids, plasma exchange (PE) and hemodialysis (HD). Further, 38 patients had died and 60 had survived within 100 days of hospitalization. Serum creatinine (Cr) was higher in patients who had died versus patients who had survived (P = 0.001). Likewise, patients with white blood cells (WBC) ≥ 10(4)/µL had higher mortality rate versus patients with WBC < 10(4)/µL (P = 0.018). GMA in patients with WBC ≥ 10(4)/µL showed improved prognosis versus in patients with WBC ≥ 10(4)/µL who did not receive GMA (P = 0.0006). Corticosteroids, plasma exchange and HD did not significantly impact prognosis of SAH patients. CONCLUSIONS: Our perception is that, patients with elevated myeloid leucocytes benefit most from GMA, while plasma exchange appears to support patients with coagulation deficiency or high plasma bilirubin and HD has indication in patients with high Cr.

Inter-professional and inter-departmental alcoholism rehabilitation program.

A 3-month alcoholism rehabilitation program at psychiatric hospitals is common in Japan for patients with alcohol use disorder (AUD). However, many AUD patients are often hospitalized for the treatment of digestive disorders due to alcohol-related liver diseases and pancreatitis. In this sense, AUD patients need to be better supported by professionals and departments in general hospitals. Here we analyzed the problems in alcohol-related medical care in Japan and examined the measures to be taken at general hospitals.

Ameliorating effects of compounds derived from Salvia miltiorrhiza root extract on microcirculatory disturbance and target organ injury by ischemia and reperfusion.

Ischemia and reperfusion (I/R) exerts multiple insults in microcirculation, frequently accompanied by endothelial cell injury, enhanced adhesion of leukocytes, macromolecular efflux, production of oxygen free radicals, and mast cell degranulation. Since the microcirculatory disturbance results in injury of organ involved, protection of organ after I/R is of great importance in clinic. Salvia miltiorrhiza root has long been used in Asian countries for clinical treatment of various microcirculatory disturbance-related diseases. This herbal drug contains many active water-soluble compounds, including protocatechuic aldehyde (PAl), 3,4-dihydroxyphenyl lactic acid (DLA) and salvianolic acid B (SalB). These compounds, as well as water-soluble fraction of S. miltiorrhiza root extract (SMRE), have an ability to scavenge peroxides and are able to inhibit the expression of adhesion molecules in vascular endothelium and leukocytes. Moreover, lipophilic compounds of SMRE also prevent the development of vascular damage; NADPH oxidase and platelet aggregation are inhibited by tanshinone IIA and tanshinone IIB, respectively, and the mast cell degranulation is blunted by cryptotanshinone and 15,16-dihydrotanshinone I. Thus, the water-soluble and lipophilic compounds of SMRE appear to improve the I/R-induced vascular damage multifactorially and synergically. This review will summarize the ameliorating effect of compounds derived from SMRE on microcirculatory disturbance and target organ injury after I/R and will provide a new perspective on remedy with multiple drugs.

A new prognostic formula for adult acute liver failure using computer tomography-derived hepatic volumetric analysis.

BACKGROUND/AIMS: King's College Hospital (KCH) criteria and the model for end-stage liver disease (MELD) score are useful and widely-employed prognostic markers for acute liver failure (ALF). We previously reported that liver atrophy is an important prognostic factor for ALF. The aim of the present study was to assess the value of liver volumetry and to generate a new prognostic formula. METHODS: Computed tomography-derived liver volume (CTLV) and standardized liver volume (SLV) of 30 adult ALF patients were calculated at the time of diagnosis. Patients were assigned to one of two groups: group A consisted of 13 patients who recovered without surgical intervention, and group B consisted of 17 patients who died due to liver failure or who underwent living donor liver transplantation (LDLT). RESULTS: The median CTLV/SLV ratios of groups A and B were 1.019 and 0.757, respectively (P = 0.0009). The difference was most significant (P = 0.0002) at the probability cutoff point of 0.80 for CTLV/SLV ratio; the sensitivity and specificity were 76.5% and 92.3%, respectively. Serum total bilirubin (TB) levels and CTLV/SLV ratio were selected as independent prognostic factors by multivariate analysis. A prognostic formula including volumetric analysis was established: Z = -2.3813 - [0.15234 x TB (mg/dl)] + [4.5734 x CTLV/SLV] (AUC = 0.87783, P = 0.0002). CONCLUSIONS: The CTLV/SLV ratio is a very useful marker for predicting the prognosis of adult ALF. Our prognostic formula including only the CTLV/SLV ratio and TB is simple and useful and awaits validation in a future larger-scale prospective study.

[Influence of habitual drinking and viral hepatitis type C in the progression of liver cirrhosis].

Recently prevalence of alcoholic liver disease has been increasing in Japan associated with an increase in alcoholic beverage consumption. In the present study, we addressed the recent trend in the etiology of liver cirrhosis (LC) in Japan, and investigated the influence of habitual drinking and viral hepatitis type C in the progression of LC. We carried out nation-wide survey by asking for the hospitals that are approved by the Japanese Society of Gastroenterology for the etiology of in-patients with LC, and compared to that in our hospital. Regarding the cases in nation-wide survey, 1274 cases (14%) of 9126 patients with LC were pure (without any markers of hepatitis virus) heavy drinkers, and 580 cases (6%) were heavy drinkers with any markers of hepatitis virus. However, in our general hospital, 24 cases of 101 patients with LC (24%) were pure heavy drinker, and 31 cases (30%) were heavy drinkers with any markers of hepatitis virus. In conclusion, although influence of hepatitis virus infection in alcoholic LC has been decreasing, it still plays an important role in the progression of alcoholic LC, especially in the general hospitals. Education of abstinence or low risk drinking is important not only heavy drinkers but also habitual drinkers with hepatitis virus infection.

Effect of a herbal medicine on fatty liver in rats fed ethanol chronically.

AIM: The objective of this study was to determine whether Cardiotopic Pills (CP) affects fatty liver in rats fed ethanol chronically. MATERIALS AND METHODS: Male Wistar rats were treated with liquid diet that contained ethanol (36% of total calories) or an isocaloric carbohydrate instead of ethanol for 6 weeks. CP, an oral herbal medicine including Danshen (Salviae Miltiorrhiza), Panax notoginseny and Dyroblanops aromatica gaertn, have been clinically used for vascular diseases such as coronary diseases and cerebral infarction. CP was administered orally with the liquid diets for 2 weeks 0.4 mg/kg body weight/day with the liquid diet thereafter. Serum triglyceride and total cholesterol levels, total protein, albumin, and AST and ALT activities are measured. Histological examination was also carried out. In another set of experiments, autofluorescence of NAD(P)H, an indicator of mitochondrial O2 consumption and redox status, was measured by an intravital microscopy, and peroxisome proliferators-activated receptor-(PPAR)-alpha and gamma mRNA levels were evaluated by real time quantitative PCR methods. RESULTS: Chronic ethanol consumption elevated serum triglyceride level, and caused fatty degeneration of liver. After administration of CP, fatty degeneration was not observed in rats fed ethanol chronically. Elevation of serum triglyceride level was not noted after treatment with CP (Ethanol: 79.4 +/- 9.3 mg/dl, Ethanol+CP: 48.0 +/- 4.4, respectively, p<0.05). CP did not affect any other laboratory data or NAD(P)H levels. Chronic ethanol consumption did not affect PPAR-gamma mRNA levels, while it decreased PPAR-alpha mRNA levels in the liver. CP prevented the ethanol-induced decrease in PPAR-alpha mRNA levels. CP and its components could enhance expression of PPAR-alpha mRNA levels. CONCLUSION: These results suggest that CP may be useful to prevent alcoholic fatty liver via enhanced expression of PPAR-alpha.

Protective effects of dihydroxylphenyl lactic acid and salvianolic acid B on LPS-induced mesenteric microcirculatory disturbance in rats.

Salvia miltiorrhiza is a Chinese medicine widely used for treatment of various cardiovascular diseases. However, little is known about the role of dihydroxylphenyl lactic acid (DLA) and salvianolic acid B (SAB), the main ingredients of S. miltiorrhiza, in the microcirculation. This study aimed to investigate the effect of DLA and SAB on LPS-elicited microcirculatory disturbance, focusing especially on leukocyte adhesion and its potential mechanism. Mesenteric venular diameter, velocity of red blood cells in venules, shear rate of the venular wall, numbers of leukocytes adherent to and emigrated across the venular wall, and mast cell degranulation were determined by an inverted microscope in rats after LPS infusion with or without DLA or SAB. Expression of CD11b and CD18 and production of superoxide anion (*O2-) and hydrogen peroxide (H2O2) by neutrophils were evaluated in vitro by flow cytometry. LPS exposure induced a significant increase in the number of adherent and emigrated leukocytes and mast cell degranulation, and a prominent decrease in the velocity of red blood cells in venules and shear rate of the venular wall. Additionally, in vitro experiments revealed an apparent enhancement in expression of CD11b and CD18 and production of *O2- and H2O2 by rat neutrophils by LPS stimulation. Treatment with DLA or SAB significantly ameliorated LPS-induced microcirculatory disturbance in rat mesentery and inhibited both the expression of CD11b and CD18 and the production of *O2- and H2O2 by neutrophils caused by LPS.

Different socioeconomic backgrounds between hazardous drinking and heavy episodic drinking: Prevalence by sociodemographic factors in a Japanese general sample.

BACKGROUND: Hazardous drinking (HD) and heavy episodic drinking (HED) constitute different types of alcohol-related harm. The socioeconomic status (SES) background of various alcohol consumption behaviors is not clear. The purpose of this study was to clarify existing SES differences between HD and HED. METHODS: The 2013 national survey regarding alcohol use among Japanese adults was utilized. The results from 1193 men and 1503 women aged 20-64 years were included in the analysis. Education attainment, household income, marital status, working status, and occupation were adopted as SES determinants. Binomial logistic regression analysis was conducted to estimate the odds ratios (ORs) of HD and HED for each SES group. RESULTS: ORs (95% confidence intervals) of HD were higher among persons with less education among both men [1.61 (1.18-2.20)] and women [1.78 (1.19-2.67)]. The OR of HED in men was significantly higher among those who belonged to high household income, were married, and managers or professionals. The OR of HED among women was higher in persons who were employed, as compared with those who engaged in housework. There were no correlations between HED and educational background. CONCLUSIONS: This study showed that in Japan, a lower educational background for both men and women was associated with a higher risk for HD, while higher current SES for men and working women were associated with a higher risk for HED. It is necessary to recognize the SES differences between HD and HED to achieve a policy to reduce alcohol-related harm.

Protective effects of ginsenoside Rb1, ginsenoside Rg1, and notoginsenoside R1 on lipopolysaccharide-induced microcirculatory disturbance in rat mesentery.

Ginsenoside Rb1 (Rb1), ginsenoside Rg1 (Rg1), and notoginsenoside R1 (R1) are major active components of Panax notoginseng, a Chinese herb that is widely used in traditional Chinese medicine to enhance blood circulation and dissipate blood stasis. To evaluate the effect of these saponins on microcirculatory disturbance induced by lipopolysaccharide (LPS), vascular hemodynamics in rat mesentery was observed continuously during their administration using an inverted microscope and a high speed video camera system. LPS administration decreased red blood cell velocity but Rb1, Rg1, and R1 attenuated this effect. LPS administration caused leukocyte adhesion to the venular wall, mast cell degranulation, and the release of cytokines. Rb1, Rg1, and R1 reduced the number of adherent leukocytes, and inhibited mast cell degranulation and cytokine elevation. In vitro experiments using flow cytometry further demonstrated that a) the LPS-enhanced expression of CD11b/CD18 by neutrophils was significantly depressed by Rb1 and R1, and b) hydrogen peroxide (H(2)O(2)) release from neutrophils in response to LPS stimulation was inhibited by treatment with Rg1 and R1. These results suggest that the protective effect of Rb1 and R1 against leukocyte adhesion elicited by LPS may be associated with their suppressive action on the expression of CD11b/CD18 by neutrophils. The protective effect against mast cell degranulation by Rb1 and R1, and the blunting of H(2)O(2) release from neutrophils by Rg1 and R1 suggest mechanistic diversity in the effects of Panax notoginseng saponins in the attenuation of microcirculatory disturbance induced by LPS.

Compound Danshen injection improves endotoxin-induced microcirculatory disturbance in rat mesentery.

AIM: To investigate the effect of compound Danshen injection on lipopolysaccharide (LPS)-induced rat mesenteric microcirculatory dysfunctions and the underlying possible mechanism by an inverted intravital microscope and high-speed video camera system. METHODS: LPS was continuously infused through the jugular artery of male Wistar rats at the dose of 2 mg/kg per hour. Changes in mesenteric microcirculation, such as diameters of arterioles and venules, velocity of RBCs in venules, leukocyte rolling, adhesion and emigration, free radicals released from post-capillary venules, FITC-albumin leakage and mast cell degranulation, were observed through an inverted intravital microscope assisted with CCD camera and SIT camera. Meanwhile, the expression of adhesion molecules CD11b/CD18 and the production of free radical in neutrophils, and the expression of intercellular adhesion molecule 1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs) were quantified by flow cytometry (FACS) in vitro. RESULTS: The continuous infusion with LPS resulted in a number of responses in microcirculation, including a significant increase in the positive region of venule stained with Monastral blue B, rolling and adhesion of leukocytes, production of oxygen radical in venular wall, albumin efflux and enhanced mast cell degranulation in vivo, all of which, except for the leukocyte rolling, were attenuated by the treatment with compound Danshen injection. Experiments performed in vitro further revealed that the expression of CD11b/CD18 and the production of oxygen free radical in neutrophils, and the expression of ICAM-1 in HUVECs were increased by exposure to LPS, and they were attenuated by compound Danshen injection. CONCLUSION: These results suggest that compound Danshen injection is an efficient drug with multi-targeting potential for improving the microcirculatory disturbance.

Attenuating effect of Myakuryu on mesenteric microcirculatory disorders induced by ischemia and reperfusion.

Myakuryu (MR) is a newly developed herbal medicine composing Crataegue oinnatifida bge (COB), Panax notoginseng (PN) and Ginkyo biloba (GB). To examine the effectiveness of MR, we investigated its effects on rat mesenteric microcirculatory injury induced by ischemia/reperfusion (I/R). The mesenteric microcirculation of ileocecal portion of a male Wistar rat was observed through an inverted-type intravital microscope assisted with a charge-coupled devise (CCD) camera. Mesenteric I/R was conducted by a ligation of the mesenteric artery and vein (10 min) and subsequent release of the occlusion. We measured venular diameter, the number of adherent leukocytes, dihydrorhodamine 123 (DHR) fluorescence as an indicator of oxidative stress and mast cell degranulation, with or without MR extract (0.4 g/kg b.w.) via an orogastric tube 1 hr before I/R. The diameters of the observed mesenteric venules were not changed after the mesenteric I/R. MR had no effect on venular diameter. The leukocytes adhering to the post-capillary venular walls started just after reperfusion, and increased thereafter. The increased number of adherent leukocytes was significantly reduced by treatment with MR. DHR fluorescence ratio was significantly increased along the venular wall. MR attenuated the increased oxidation. The mesenteric I/R induced mast cell degranulation. The increase in mast cell degranulation was inhibited by MR. In conclusion, oral administration of MR attenuates I/R-induced microvascular damages in rat mesentery. MR has a therapeutic potential for prevention of I/R-related microvascular injury.