[[39]] For important considerations related to performing surgical

procedures in persons with hemophilia, please see “Surgery and Invasive Procedures”. Specific issues in relation to orthopedic surgery include: Orthopedic surgeons should have had specific training Tamoxifen in surgical management of persons with hemophilia. [[3]] Performing multiple site elective surgery in a simultaneous or staggered fashion to use clotting factor concentrates judiciously should be considered. (Level 3) [[50]50] Local coagulation enhancers may be used. Fibrin glue is useful to control oozing when operating in extensive surgical fields. (Level 3) [[36, 51, 52]36,51,52] Postoperative care in patients with hemophilia requires closer monitoring of pain and often higher doses of analgesics in the immediate postoperative period. (Level 5) [[36]] Good communication with the postoperative rehabilitation selleck kinase inhibitor team is essential [[39]]. Knowledge of the details of the surgery performed and intra-operative joint status will facilitate planning of an appropriate rehabilitation program. Postoperative rehabilitation should be carried out by a physiotherapist experienced in hemophilia management. Rehabilitation may have to progress more slowly in persons with hemophilia. Adequate pain control is essential to allow appropriate exercise and mobilization. These

principles also apply to fixation of fractures and excision of pseudotumors. Inhibitors” in hemophilia refer to IgG antibodies that neutralize clotting factors. In the current era in which clotting factor concentrates have been subjected to appropriate viral inactivation, inhibitors to FVIII MCE or FIX are considered the most severe treatment-related complication in hemophilia. The presence of a new inhibitor should be suspected in any patient who fails to respond clinically to clotting factors, particularly if he has been previously responsive. In this situation, the expected recovery and half-life of the transfused clotting factor are severely diminished. Inhibitors are more frequently encountered

in persons with severe hemophilia compared with those with moderate or mild hemophilia. The cumulative incidence (i.e., lifetime risk) of inhibitor development in severe hemophilia A is in the range of 20–30% and approximately 5–10% in moderate or mild disease. [[53, 54]] In severe hemophilia A, the median age of inhibitor development is 3 years or less in developed countries. In moderate/mild hemophilia A, it is is closer to 30 years of age, and is often seen in conjunction with intensive FVIII exposure with surgery. [[55, 56]] In severe hemophilia, inhibitors do not change the site, frequency, or severity of bleeding. In moderate or mild hemophilia, the inhibitor may neutralize endogenously synthesized FVIII, thereby effectively converting the patient’s phenotype to severe.

The few studies on wine and headache were mostly presented as abstracts despite the common knowledge and patients’ complaints about wine ingestion and headache attacks. These studies suggest that red wine, but not white and sparkling

wines, do trigger headache and migraine attacks independently of dosage in less than 30% of the subjects. Wine, and specifically red wine, is a migraine trigger. Non-migraineurs may have headache attacks with wine ingestion as well. The reasons for that triggering potential are uncertain, but the presence of phenolic flavonoid radicals and the potential for interfering with the central serotonin PF-02341066 price metabolism are probably the underlying mechanisms of the relationship between wine and headache. Further controlled studies www.selleckchem.com/products/3-deazaneplanocin-a-dznep.html are necessary to enlighten this traditional belief. The idea of dietary migraine or the triggering of migraine attacks with food and beverages has long been disseminated. In 1778, Fothergill first described headache attacks after the ingestion of specific dietary factors, but the variability of clinical presentations among and within migraine and non-migraine sufferers has cast doubts about the real existence of such entity.[1, 2] Particularly with regards to wine, medicine has been imaginative in correlating its consumption

with bad and good consequences throughout the centuries, with the first references about a possible relationship between wine and medicine in Mesopotamia 7000 years B.C.[3, 4] When wine making arrived in ancient Greece, it was enjoyed by the whole spectrum of society, and became a popular MCE公司 theme in literature, religion, leisure, medicine, and mythology. Hippocrates promoted wine as part of a healthy diet. He also claimed that wine was good for disinfecting wounds, as well as a liquid in which medications could be mixed and taken more easily by patients. Hippocrates said wine

should be used to alleviate pain during childbirth, for symptoms of diarrhea, and even lethargy.3-5 Around 1863, a French Corsican chemist called Angelo Mariani developed a beverage containing Bordeaux wine and cocaine (approximately 6 mg of cocaine per fluid ounce of wine). This beverage named Vin Tonique Mariani was suggested as a substitute for opiates and was awarded a Vatican Gold Medal by Pope Leo XIII in addition to an endorsement of its use.[6] Wine’s intrinsic link with the practice of medicine was also featured prominently in the first printed book on wine written by Arnaldus de Villa Nova (circa. 1235-1311 A.D.), a physician, who wrote at length on wine’s benefits for the treatment of many illnesses and conditions, including sinus problems and dementia.[5] For triggering migraine and/or headache attacks, red wine is well known as a trigger and has been so since antiquity when Celsus (25 B.C.-50 A.D.) described pain contracted by drinking wine. Six centuries later, Paul of Aegina (625-690 A.D.

Cirrhosis was diagnosed on the basis of Selleckchem PD0325901 clinical, laboratory, and ultrasound evidence. Criteria for exclusion were the use of drugs known to interfere with blood coagulation, ongoing bacterial

infections, hepatocellular carcinoma, and extrahepatic malignancy. The severity of the disease was estimated according to Child-Turcotte-Pugh score.11 One-hundred-five healthy individuals matched with the patient population for age and sex, were enrolled in this study as controls. In addition to the above controls, a population of 37 individuals (4 with and 33 without a previous history of thrombosis) known to be carriers of the factor V Leiden mutation was included for comparative purposes. Blood for laboratory analyses was drawn by clean venipuncture and collected in vacuum tubes (Becton Dickinson, Meylan, France) containing 109 mM trisodium citrate as anticoagulant in the proportion of 1/9 parts of anticoagulant/blood. Blood was centrifuged within 30 minutes from collection at 2880g for 15 minutes at room temperature. Platelet-free plasma was then harvested, quick frozen in liquid nitrogen and stored at −70°C until tested. This is a commercially available chromogenic assay (HemosIL Thrombopath;

Instrumentation Laboratory, Orangeburg, NY) designed to globally evaluate the functionality of the protein C anticoagulant system.12 It is based on the ability of endogenous activated protein C generated after activation of protein C by a snake venom extract (Protac) to reduce the tissue factor–induced thrombin generation. The amount of thrombin is evaluated by recording changes in optical CX-4945 ic50 density (OD) at 405 nm in the presence (A) or absence (B) of Protac after adding a thrombin-specific chromogenic 上海皓元 substrate. The assay kit contains all the reagents that are needed to run the test. All reagents, except for the diluent are lyophilized and were reconstituted with distilled water before use according to

the manufacturer’s specification. Briefly, 10 μL of the plasma sample and 40 μL of the Thrombopath diluent were incubated with either the Thrombopath activator A or the Thrombopath activator B (45 μL) for 120 seconds, before the Thrombopath thromboplastin reagent (50 μL) was added. After 90 seconds of incubation, 50 μL of the Thrombopath substrate was added, and change in OD at 405 nm was recorded for 45 seconds. Results are expressed as the Protac-induced coagulation inhibition percentage (PICI%) calculated by the following equation: PICI% = ([B − A]/B) × 100, where B and A are the OD for plasma tested in the absence (B) or presence (A) of Protac. The smaller the PICI% value, the greater the procoagulant imbalance. Testing with Thrombopath was performed by a fully automated coagulation analyzer (ACL 10000; Instrumentation Laboratory, Bedford, MA) according to the manufacturer’s specification. To minimize methodological variability, equal numbers of plasmas from patients and controls were included on each test occasion.

Cirrhosis was diagnosed on the basis of Temozolomide datasheet clinical, laboratory, and ultrasound evidence. Criteria for exclusion were the use of drugs known to interfere with blood coagulation, ongoing bacterial

infections, hepatocellular carcinoma, and extrahepatic malignancy. The severity of the disease was estimated according to Child-Turcotte-Pugh score.11 One-hundred-five healthy individuals matched with the patient population for age and sex, were enrolled in this study as controls. In addition to the above controls, a population of 37 individuals (4 with and 33 without a previous history of thrombosis) known to be carriers of the factor V Leiden mutation was included for comparative purposes. Blood for laboratory analyses was drawn by clean venipuncture and collected in vacuum tubes (Becton Dickinson, Meylan, France) containing 109 mM trisodium citrate as anticoagulant in the proportion of 1/9 parts of anticoagulant/blood. Blood was centrifuged within 30 minutes from collection at 2880g for 15 minutes at room temperature. Platelet-free plasma was then harvested, quick frozen in liquid nitrogen and stored at −70°C until tested. This is a commercially available chromogenic assay (HemosIL Thrombopath;

Instrumentation Laboratory, Orangeburg, NY) designed to globally evaluate the functionality of the protein C anticoagulant system.12 It is based on the ability of endogenous activated protein C generated after activation of protein C by a snake venom extract (Protac) to reduce the tissue factor–induced thrombin generation. The amount of thrombin is evaluated by recording changes in optical www.selleckchem.com/screening/chemical-library.html density (OD) at 405 nm in the presence (A) or absence (B) of Protac after adding a thrombin-specific chromogenic MCE substrate. The assay kit contains all the reagents that are needed to run the test. All reagents, except for the diluent are lyophilized and were reconstituted with distilled water before use according to

the manufacturer’s specification. Briefly, 10 μL of the plasma sample and 40 μL of the Thrombopath diluent were incubated with either the Thrombopath activator A or the Thrombopath activator B (45 μL) for 120 seconds, before the Thrombopath thromboplastin reagent (50 μL) was added. After 90 seconds of incubation, 50 μL of the Thrombopath substrate was added, and change in OD at 405 nm was recorded for 45 seconds. Results are expressed as the Protac-induced coagulation inhibition percentage (PICI%) calculated by the following equation: PICI% = ([B − A]/B) × 100, where B and A are the OD for plasma tested in the absence (B) or presence (A) of Protac. The smaller the PICI% value, the greater the procoagulant imbalance. Testing with Thrombopath was performed by a fully automated coagulation analyzer (ACL 10000; Instrumentation Laboratory, Bedford, MA) according to the manufacturer’s specification. To minimize methodological variability, equal numbers of plasmas from patients and controls were included on each test occasion.

Cirrhosis was diagnosed on the basis of buy AZD2281 clinical, laboratory, and ultrasound evidence. Criteria for exclusion were the use of drugs known to interfere with blood coagulation, ongoing bacterial

infections, hepatocellular carcinoma, and extrahepatic malignancy. The severity of the disease was estimated according to Child-Turcotte-Pugh score.11 One-hundred-five healthy individuals matched with the patient population for age and sex, were enrolled in this study as controls. In addition to the above controls, a population of 37 individuals (4 with and 33 without a previous history of thrombosis) known to be carriers of the factor V Leiden mutation was included for comparative purposes. Blood for laboratory analyses was drawn by clean venipuncture and collected in vacuum tubes (Becton Dickinson, Meylan, France) containing 109 mM trisodium citrate as anticoagulant in the proportion of 1/9 parts of anticoagulant/blood. Blood was centrifuged within 30 minutes from collection at 2880g for 15 minutes at room temperature. Platelet-free plasma was then harvested, quick frozen in liquid nitrogen and stored at −70°C until tested. This is a commercially available chromogenic assay (HemosIL Thrombopath;

Instrumentation Laboratory, Orangeburg, NY) designed to globally evaluate the functionality of the protein C anticoagulant system.12 It is based on the ability of endogenous activated protein C generated after activation of protein C by a snake venom extract (Protac) to reduce the tissue factor–induced thrombin generation. The amount of thrombin is evaluated by recording changes in optical BVD-523 density (OD) at 405 nm in the presence (A) or absence (B) of Protac after adding a thrombin-specific chromogenic medchemexpress substrate. The assay kit contains all the reagents that are needed to run the test. All reagents, except for the diluent are lyophilized and were reconstituted with distilled water before use according to

the manufacturer’s specification. Briefly, 10 μL of the plasma sample and 40 μL of the Thrombopath diluent were incubated with either the Thrombopath activator A or the Thrombopath activator B (45 μL) for 120 seconds, before the Thrombopath thromboplastin reagent (50 μL) was added. After 90 seconds of incubation, 50 μL of the Thrombopath substrate was added, and change in OD at 405 nm was recorded for 45 seconds. Results are expressed as the Protac-induced coagulation inhibition percentage (PICI%) calculated by the following equation: PICI% = ([B − A]/B) × 100, where B and A are the OD for plasma tested in the absence (B) or presence (A) of Protac. The smaller the PICI% value, the greater the procoagulant imbalance. Testing with Thrombopath was performed by a fully automated coagulation analyzer (ACL 10000; Instrumentation Laboratory, Bedford, MA) according to the manufacturer’s specification. To minimize methodological variability, equal numbers of plasmas from patients and controls were included on each test occasion.

Intestinal decontamination with non-absorbable antibiotics restored eubiosis, decreased intestinal inflammation and permeability, and reduced

alcoholic liver disease in mice suggesting that alcohol-associated dysbi-osis induces intestinal inflammation and leakiness. To further define the role of TNFα in mediating intestinal barrier dysfunction following alcohol administration, we focused on the main receptor for TNFα, TNF-receptor 1 (TNFR-1). TNFR-1 mutant mice (TNFR-1flxneo/flxneo) were protected from intestinal barrier dysfunction as evidenced by higher levels of fecal albumin and decreased hepatic contents of bacterial products. TNFR-1 mutant mice had less liver disease as shown by lower plasma ALT levels and hepatic triglycerides. To investigate Everolimus whether TNFR-1 on intestinal epithelial cells mediates intestinal barrier dysfunction and liver disease, a functional TNFR-1 was selectively expressed click here on intestinal epithelial cells

by crossing TNFR-1flxneo/flxneo mice with Villin-Cre transgenic mice. Reactivation of TNFR-1 on enterocytes resulted in increased intestinal permeability and liver disease that is similar to wild type mice after alcohol feeding, suggesting that enteric TNFR-1 promotes intestinal barrier dysfunction and mediates alcoholic liver disease. Myosin light chain kinase (MLCK) is a downstream target of TNFα and was phosphorylated in intestinal epithelial cells following alcohol administration. Intestinal barrier loss was reduced in MCE公司 MLCK−/− mice after ch ronic alcohol feeding. While hepatic steatosis was lower in MLCK−/− mice as compared with wild type mice, liver injury was not reduced suggesting that intestinal MLCK partially

contributes to intestinal leakiness and liver disease. Conclusion: Dysbiosis-induced intestinal inflammation and TNFR-1 signaling in intestinal epithelial cells are mediating a disruption of the intestinal barrier following chronic alcohol feeding. Therefore, intestinal TNFR-1 is a crucial mediator of alcoholic liver disease. Disclosures: Samuel B. Ho – Consulting: Genentech; Grant/Research Support: Roche The following people have nothing to disclose: Peng Chen, Peter Starkel, Jerrold R. Turner, Bernd Schnabl Alcoholic liver disease is a major health issue worldwide, but effective therapies are currently unavailable. The present study tested the efficacy of Alda-1, an activator of aldehyde dehydrogenase 2, in treating alcoholic liver disease. Male C57BL/6J mice were exposed to alcohol for up to 8 weeks for a time-course study on aldehyde metabolism. The effects of Alda-1 on aldehyde dehydrogenase 2 and aldehyde clearance were determined after acute alcohol intoxication. Mice were exposed to alcohol for 8 weeks with or without Alda-1 administration for the last 10 days to test the therapeutic potential of Alda-1.

[41] Moreover, studies from the laboratory of Dr. Joel Linden demonstrated that activation of Adora2a receptors on inflammatory cells—particularly on natural killer T-cells—are

involved in liver protection from ischemia.[10] In contrast to these studies, the present findings implicate Adora2b EPZ-6438 in vitro in ENT-mediated liver protection from ischemia. Consistent with these findings, several previous studies had implicated Adora2b in tissue protection from ischemia.[24, 35-37, 42-45] In addition, it is conceivable that the timing of the injury model may contribute to such differences; while early on (e.g., 2 hours after reperfusion) the dominant protective pathway could involve Adora2b, later inflammatory changes (particularly involving T-cells) could be attenuated by Adora2a. Several studies have demonstrated that while adenosine signaling through Adora2b may be beneficial in an acute setting, this adenosine protection can become detrimental when it is prolonged.[46-49] Indeed, studies in a chronic liver disease model have shown detrimental effects of Adora2b signaling, using fatty liver disease—commonly associated with alcohol ingestion and abuse—as a model.[50, 51] During ethanol metabolism, adenosine is generated by the enzyme ecto-5′-nucleotidase, and adenosine production and adenosine receptor activation are known to

play critical roles in the development Alvelestat mw of hepatic fibrosis. Dr. Cronstein’s laboratory team therefore investigated whether adenosine and its receptors play a role in the development of alcohol-induced fatty liver. Wild-type mice fed ethanol on the Lieber-DeCarli diet developed hepatic steatosis, including increased hepatic triglyceride content, while mice lacking the ecto-5′-nucleotidase CD73 or Adora1 or Adora2b receptors were protected from developing fatty liver disease. These studies indicate that adenosine generated by ethanol metabolism plays an important role in ethanol-induced hepatic steatosis by way of both Adora1 and Adora2b and suggest that targeting adenosine 上海皓元 receptors may be effective in the prevention of alcohol-induced fatty liver.[50] Hepatic ischemia and reperfusion injury significantly contributes

to morbidity and mortality of surgical patients undergoing liver transplantation. Indeed, the present studies reveal several lines of potential treatment modalities that could be used to prevent or treat hepatic ischemia and reperfusion injury. As a first line of treatment, the present studies suggest that HIF activators could be used to treat liver ischemia and reperfusion injury. Such compounds would result in repression of ENTs, thereby promoting adenosine-dependent liver protection. At the same time, these compounds would also increase extracellular adenosine production and signaling, by transcriptionally inducing enzymes that produce adenosine during ischemic conditions.[1-4] Interestingly, a recent clinical trial shows that HIF activators can be safely used in patients for the treatment of renal anemia.

Further, CSC-mediated IL-8 production leads to increased self-renewal ability, amplified endothelial tube formation in vitro and enhanced tumorigenicity in vivo. Moreover, we have also

provided evidence that the preferential expression of IL-8 in CD133+ liver Z-VAD-FMK order CSCs is mediated through a neurotensin-activated mitogen-activated protein kinase (MAPK)-signaling cascade (Tang et al., unpubl. data, 2011 [manuscript submitted]). The identification of novel therapeutic targets for HCC treatment has begun in earnest in the field of basic liver cancer research. Although there has been a significant improvement in the detection and treatment of early stage HCC, the disease remains largely incurable because Selleck H 89 the current therapeutic regimen is unable to provide a lasting cure for patients with advanced HCC. Recent findings in the identification (Table 2) and characterization of liver CSCs have lent insight and offered great promise for developing better therapeutic strategies against the disease. CD90+CD44+ HCC cells, as discussed previously, possess a high tumorigenic capacity.23 Researchers who have characterized this

subpopulation of cells have also examined the potential benefits of targeting CD44 via a neutralizing antibody approach. The systemic administration of anti-human CD44 antibodies in immunodeficient mice, formed by the intrahepatic inoculation of CD90+ liver CSCs, suppressed tumor nodule formation in the liver and metastatic lesions in the lung.23 Furthermore, the administration of CD44 antibodies was also shown to induce apoptosis in both CD90+ and CD90- cells in vitro.23 In addition to CD44, CD133 has also been suggested as a putative therapeutic target in HCC.46 Using a murine anti-human

CD133 antibody conjugated to the cytotoxic drug, monomethyl auristatin F, Smith et al. found that the antibody-drug conjugate was able to productively induce the inhibition of CD133+ liver CSC-driven cancer cell growth both in vitro and in vivo.46 The granulin-epithelin precursor (GEP), which has been suggested to play a role in medchemexpress liver cancer cell chemoresistance,33 has also been identified as a potential target for antibody therapy.47 Indeed, anti-GEP monoclonal antibody treatment has resulted in the inhibition of tumor growth in immunodeficient mice, decreased serum GEP levels and reduced tumor angiogenesis.33 The recent work by Haraguchi et al. on the study of CD13+ liver CSCs has also demonstrated that CD13 inhibition by a CD13-neutralizing antibody could elicit cellular apoptosis and inhibit the proliferation of CD13+ liver CSCs-driven HCC. Further, when the CD13 inhibitor, ubenimex, is used in conjunction with the chemotherapeutic drug, 5-fluorouracil, a greater tumor regression was observed than when either agent was used alone.

4, 95% CI = 1.2-1.7). Similar results were found for tension-type headache (TTH), migraine, and non-classified headache. Subjects with insomnia-related Sotrastaurin research buy working disability had a 60% increased headache risk (RR = 1.6, 95% CI = 1.3-2.1). The RR was larger

for migraine (RR = 2.0, 95% CI = 1.3-3.1) than for TTH (RR = 1.5, 95% CI = 1.1-2.1). Insomnia at baseline was related to headache frequency at follow-up for both migraine (P trend = 0.02) and TTH (P trend < 0.001). Conclusion.— In headache-free subjects, insomnia was associated with an increased risk of headache 11 years later. The association was particularly strong for chronic headache. "
“(Headache 2010;50:779-789) Background.— Variables that are thought to precipitate migraine or tension-type headache episodes in children hitherto have only been studied using retrospective reports. As such, there is little empirical evidence to support the actual predictive association between presumed headache triggers and actual headache occurrence in children. Objective.— The present study sought to determine if fluctuations in weather, a commonly reported headache trigger in children, predict increased likelihood of headache

occurrence when evaluated using rigorous prospective methodology (“electronic momentary assessment”). see more Methods.— Twenty-five children (21 girls, 4 boys) between the ages of 8-17 years attending a new patient neurology clinic appointment and having a diagnosis of chronic migraine, chronic tension-type, or episodic migraine headache (with or without aura) participated in the study. Children completed baseline measures on headache characteristics, 上海皓元医药股份有限公司 presumed headache triggers, and mood and subsequently were trained in the use of electronic diaries to record information on headaches. Children then completed thrice daily

diaries on handheld computers for a 2-week time period (42 assessments per child) while data on weather variables (temperature, dew point temperature, barometric pressure, humidity, precipitation, and sunlight) in the child’s geographic location were recorded each time a diary was completed. Data were analyzed using multilevel models. Results.— Of the weather variables, relative humidity and presence of precipitation were significantly predictive of new headache onset, with nearly a 3-fold increase in probability of headache occurrence during times of precipitation or elevated humidity in the child’s area, b = 0.38, t(821) = 2.10, P = .04, and b = 0.02, t(821) = 2.81, P = .01, respectively. These associations remained after accounting for fluctuations in mood, and associations were not significantly stronger in children who at baseline thought that weather was a headache trigger for them. Changes in temperature, dew point temperature, barometric pressure, and sunlight were not significantly predictive of new headache episode occurrence in this sample. Conclusions.

13 Superoxide dismutase (SOD) activity in liver homogenate was determined according to the method described by Nandi and Chatterjee.14 This method is based on the ability of SOD to inhibit the auto-oxidation of pyrogallol at an alkaline pH. One unit of SOD is described as the amount of enzyme required to cause 50% inhibition of pyrogallol auto-oxidation. The glutathione (GSH) content

in the liver homogenate was determined using the method of Van Dooran et al.15 The basis of the GSH determination method is the reaction of Ellman’s reagent (5,5′-dithiobis-[2-nitrobenzoic acid]) with thiol groups of GSH at pH 8.0 to produce the yellow 5-thiol-2-nitrobenzoate anion. Glutathione S-transferase (GST) activity was determined according to the method of Habig et al.16 In this assay, GST catalyzes the conjugation of GSH with 1-chloro-2,4-dinitrobenzene, producing www.selleckchem.com/products/pexidartinib-plx3397.html a chromophore at 340 nm. The total protein contents of liver tissues were determined according to the Lowry method as modified by Peterson.17 Absorbances were recorded using a Shimadzu recording

spectrophotometer (UV-160) in all measurements. Liver cancer cell VX-809 datasheet line HepG2 were maintained in Roswell Park Memorial Institute-160 medium with 10% fetal bovine serum and 1% of 100 U/mL penicillin and 100 μg/mL streptomycin at 37°C inside a humidified incubator with 5% CO2 and 95% room air. Cells were subcultured every 4-7 days with trypsin/ethylenediamine tetraacetic acid (1:250; PAA Laboratory, Germany). Cells were treated with several concentrations of saffron extract for several time points. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole) proliferation assay was used in the HepG2 cell line to assess the effect on cell proliferation of a range of concentrations of saffron extract.

Cells (104) were plated and grown in 200 μL of growth medium in 96-well microtiter plates. After an overnight attachment period, cells were treated with varying concentrations of saffron extract (1.0, 2.0, 4.0, and 6.0 mg/mL) prepared from a 100 上海皓元医药股份有限公司 mg/mL stock solution dissolved in water. All studies were performed in triplicate and repeated three times independently. Cell growth was quantified by the ability of living cells to reduce the yellow dye MTT to a purple formazan product. Cells were incubated with MTT (Sigma) at 37°C in a humidified 5% CO2 atmosphere for 2 hours. The MTT formazan product was then dissolved in dimethylsulfoxide, and absorbance was measured at 570 nm in a microplate reader. One day before treatment, cells were seeded at a density of 1.2 × 106 cells per plate. After the indicated times, the cells were harvested by trypsin release, washed twice with phosphate-buffered saline, fixed with 70% ethanol, treated with 1% ribonuclease, and finally stained with propidium iodide (100 μg/mL final concentration).