Risk factors of developing non-arteric ischemic optic neuropathy in patients with type 2 diabetes mellitus:a single-center retrospective cohort study

AIM:To determine the risk factors and time to non-arteric ischemic optic neuropathy(NAION)occurrence among Thai type 2 diabetes mellitus(T2DM)patients.METHODS:A retrospective review of 266 newly diagnosed T2DM cases at Rajavithi Hospital between 2007 and 2016 was conducted to determine time to occurrence of NAION and evaluate associated risk factors.RESULTS:Hypertension and dyslipidemia were the most common pre-existing vascular diseases and there was a significant male predominance in the NAION group.The mean age of the NAION group was significantly higher than that of the group without NAION.A higher proportion of subjects in the NAION group had hypertension,dyslipidemia,high diastolic blood pressure,smokers,and had a small cup-to-disc ratio(CDR).Higher levels of triglycerides and lowdensity lipoprotein-cholesterol in the group with NAION.Fiftyfive patients among 266 participants(20.68%)developed NAION during a mean follow-up time of 81.26±25.04mo.In a multivariable logistic regression analysis,dyslipidemia(OR=8.36,95%CI,3.447–20.273,P<0.001),high low density lipoprotein levels(OR=1.017,95%CI,1.004–1.029,P=0.009),and small CDR(OR=11.92,95%CI,4.477–31.741,P<0.001)were significant risk factors for NAION development.Smoking was the strongest predictive risk(OR=12.843,95%CI,3.959–41.659,P<0.001).Vascular complications of T2DM and aspirin were not associated with NAION.CONCLUSION:T2DM patients with dyslipidemia or a small CDR should be carefully followed up as they are at increased risk of developing NAION.

Sensory outcome of exotropia surgery in Thailand:a retrospective multicenter study

AIM:To evaluate the characteristics of exotropia(XT)and motor-sensory outcomes after surgical correction and to determine the factors associated with sensory outcomes of XT surgery.METHODS:The medical records of all patients that were diagnosed with XT and underwent strabismus surgery in 13 major government hospitals in Thailand;from January 2012 to December 2019,were retrospectively reviewed.Univariable and multivariable logistic regression were performed to identify factors related to binocular vision.RESULTS:Data of five hundred and thirty-seven patients were analyzed.Two hundred and twenty-six patients were men(42.1%).The median age of onset was 3 years old[Interquartile range(IQR):1,8].The median age at diagnosis was 9.21 years old(IQR;4.64,21.06).intermittent exotropia[X(T)]was the most common type(52.1%);19.5%of the patients had amblyopia.For refractive error,spherical equivalent refraction on right eye(RE)and left eye(LE)were-0.53±2.45 diopters(D)(range-14.88 to+10 D)and-0.48±2.37 D(range-19.50 to+7.75 D),respectively.The mean angle of deviation at distance and near before surgery were 42.06±14.91 prism diopters(PD)and 40.81±16.09 PD,respectively.Follow-up time after first operation was 2.48±2.27y.Four hundred sixtytwo patients(86%)needed only one operation and 299(55.6%)patients had bilateral lateral rectus recession.At final visit,the mean angles of deviation at distance and near decreased to 5.76±8.96 PD and 5.01±8.73 PD,respectively.After surgery,two hundred seventy-three patients(50.8%)were evaluated for binocular function,but the others did not have result.From multivariable logistic regression in 273 patients,the factors related to better binocular function were type of XT which was X(T)[adjusted odds ratio(aOR)10.35;95%CI:4.73,22.66]compared to constant XT,without amblyopia(aOR 3.97;95%CI:1.84,8.53),underwent only single operation compared with more than 1 operation(aOR 3.80,95%CI:1.58,9.16),the angle of deviation at near in last visit less than 10 PD better than 10–30 PD with aOR 0.42(95%CI:0.18,0.96)and type of refraction revealed isometropia better than anisometropia with aOR 4.13(95%CI:1.19,14.32).CONCLUSION:The surgical outcomes of XT within one operation in Thailand is 86%.The factors related to achieve binocular function includes type of XT as X(T),without amblyopia,angle of deviation at final visit less than 10 PD,isometropia type of refraction and underwent only one surgical correction.

Roles of the MEK1/2 and AKT pathways in CXCL12/CXCR4 induced cholangiocarcinoma cell invasion

AIM： To evaluate the expression of C-X-C motif chemokine receptor 4 （CXCR4） and its signaling cascades, which were previously identified as a key factor for cancer cell progression and metastasis, in cholangiocarcinoma cell lines. METHODS： The expression of CXCR4 and its signaling cascades were determined in the cholangiocarcinoma cell lines （RMCCA1 and KKU100） by Western blotting. The invasion assays and the detection of actin polymerization were tested in these cholangiocarcinoma cells treated with CXC chemokine ligand -12 （CXCL12）. RESULTS： Expression of CXCR4 was detected in both cholangiocarcinoma cell lines and activation of CXCR4 with CXCL12 triggered the signaling via the extracellular signal-regulated kinase-1/2 （ERK1/2） and phosphoinositide 3-kinase （PI3K） and induction of cholangiocarcinoma cell invasion, and displayed high levels of actin polymerization. Addition of CXCR4 inhibitor （AMD3100） abrogated CXCL12-induced phosphorylation of MEKI/2 and Akt in these cells. Moreover, treatment with MEK1/2 inhibitor （U0126） or PI3K inhibitor （LY294002） also attenuated the effect of CXCL12- induced cholangiocarcinoma cell invasion. CONCLUSION： These results indicated that the activation of CXCR4 and its signaling pathways （MEK1/2 and Akt） are essential for CXCL12-induced cholangiocarcinoma cell invasion. This rises Implications on a potential role for the inhibition of CXCR4 or its signal cascades in the treatment of cholangiocarcinoma.

Ligation of intersphincteric fistula tract and its modification: Results from treatment of complex fistula

AIM: To compare healing rates between intersphincteric fistula tract (LIFT) and LIFT plus partial fistulectomy procedures. METHODS: A study of complex fistula-in-ano patients was carried out from 1 st March 2010 to 31 th January 2012. All operations were done by colorectal surgeons at a referral center in a Ministry of Public Health hospital. Data collected included patients' demographic details, fistula type determined by endorectal-ultraso-nography, preoperative and postoperative continence status, previous operations, time between diagnosis of fistula-in-ano and operation, type of surgery, healing rates, recurrence rates, and types of failure examined by endorectal-ultrasosnography, re-operation in recurrence or failure cases, and complications. RESULTS: The study involved 41 patients whose average age was 40.78 ± 11.84 years (range: 21-71 years). The major fistula type was high-transsphincteric type fistula. The median follow-up period was 24 wk. The overall success rate was 83%: in the LIFT (Ligation intersphincteric fistula tract) group the success rate was 81% and in the LIFT plus (LIFT with partial coreout fistulectomy) group it was 85% (P = 0.529). The median wound-healing time was 4 wk in both groups (P = 0.262). The median time to recurrence was 12 wk. Neither group had incontinence (Wexner incontinence score-0) and the difference in healing rates between the two groups was not statistically significant. CONCLUSION: There was no difference in results between LIFT and LIFT plus operations. The LIFT procedure is a good option for maintaining continence in management of fistula-in-ano.

Malaria parasite carbonic anhydrase:inhibition of aromatic/heterocyclic sulfonamides and its therapeutic potential

Plasmodium falciparum(P.falciparum) is responsible for the majority of life-threatening cases of human malaria,causing 1.5-2.7 million annual deaths.The global emergence of drug-resistant malaria parasites necessitates identification and characterisation of novel drug targets and their potential inhibitors.We identified the carbonic anhydrase(CA) genes in P.falciparum.The pfGA gene encodes an α-carbonic anhydrase,a Zn^(2+)-metalloenzme,possessing catalytic properties distinct from that of the human host CA enzyme.The amino acid sequence of the pfCA enzyme is different from the analogous protozoan and human enzymes.A library of aromatic/heterocyclic sulfonamides possessing a large diversity of scaffolds were found to be very good inhibitors for the malarial enzyme at moderate-low micromolar and submicromolar inhibitions.The structure of the groups substituting the aromatic-ureido-or aromatic-azomethine fragment of the molecule and the length of the parent sulfonamide were critical parameters for the inhibitory properties of the sulfonamides.One derivative,that is,4-(3,4-dichlorophenylureido)thioureidobcnzcnesulfonamide(compound 10) was the most effective in vitro Plasmodium falciparum CA inhibitor,and was also the most effective antimalarial compound on the in vitro P.falciparum growth inhibition.The compound 10 was also effective in vivo antimalarial agent in mice infected with Plasmodium berghei,an animal model of drug testing for human malaria infection. It is therefore concluded that the sulphonamide inhibitors targeting the parasite CA may have potential for the development of novel therapies against human malaria.

Quantitative analysis of γ-oryzanol content in cold pressed rice bran oil by TLC-image analysis method

Objective:To develop and validate an image analysis method for quantitative analysis ofγ-oryzanol in cold pressed rice bran oil.Methods:TLC-densitometric and TLC-image analysis methods were developed,validated,and used for quantitative analysis of γ-oryzanol in cold pressed rice bran oil.The results obtained by these two different quantification methods were compared by paired t-test.Results:Both assays provided good linearity,accuracy,reproducibility and selectivity for determination of γ-oryzanol.Conclusions:The TLC-densitomelric and TLC-image analysis methods providett a similar reproducibility,accuracy and selectivity for the quantitative determination of γ-oryzanol in cold pressed rice bran oil.A statistical comparison of the quantitative determinations of γ-oryzanol in samples did not show any statistically significant difference between TLC-densitometric and TLC-image analysis methods.As both methods were found to be equal,they therefore can be used for the determination of γ-oryzanol in cold pressed rice bran oil.

Bacterial infections in cirrhosis: A critical review and practical guidance

Bacterial infection is common and accounts for major morbidity and mortality in cirrhosis. Patients with cirrhosis are immunocompromised and increased susceptibility to develop spontaneous bacterial infections, hospital-acquired infections, and a variety of infections from uncommon pathogens. Once infection develops, the excessive response of pro-inflammatory cytokines on a pre-existing hemodynamic dysfunction in cirrhosis further predispose the development of serious complications such as shock, acute-on-chronic liver failure, renal failure, and death. Spontaneous bacterial peritonitis and bacteremia are common in patients with advanced cirrhosis, and are important prognostic landmarks in the natural history of cirrhosis. Notably, the incidence of infections from resistant bacteria has increased significantly in healthcare-associated settings. Serum biomarkers such as procalcitonin may help to improve the diagnosis of bacterial infection. Preventive measures(e.g., avoidance, antibiotic prophylaxis, and vaccination), early recognition, and proper management are required in order to minimize morbidity and mortality of infections in cirrhosis.

Prospective study of MMP7 serum levels in the diagnosis of cholangiocarcinoma

AIM: To determine whether the serum level of matrix metalloproteinase-7 (MMP7) has the potential to diagnosis cholangiocarcinoma from benign biliary tract diseases. METHODS: This study was performed according to the PRoBE (a prospective-specimen-collection, retrospectiveblinded-evaluation) design. A total of 187 patients with obstructive jaundice were consecutively enrolled. After the diagnostic status of these patients was ascertained, their levels of serum MMP7 were assayed and compared with serum carbohydrate antigen 19-9 (CA19-9). This was conducted in a blinded case (cholangiocarcinoma)control (benign biliary tract disease) setup. RESULTS: MMP7 and CA19-9 serum levels were significantly elevated in cholangiocarcinoma patients (P < 0.001). The area under the curve (AUC) from a receiver operating characteristic (ROC) curve analysis for thediagnosis of cholangiocarcinoma, using MMP7 was more accurate than CA19-9 (AUC = 0.84, 95% CI: 0.778-0.903 for MMP7 and AUC = 0.79, 95% CI: 0.708-0.868 for CA19-9). The sensitivity and specificity of serum MMP7 (cut-off value of 5.5 ng/mL) was 75% and 78%, respectively, while the sensitivity and specificity of serum CA19-9 (cut-off value of 100 U/mL) was 68% and 87%, respectively. CONCLUSION: Serum values of MMP7 and CA19-9 appear to be useful biomarkers for differentiating cholangiocarcinoma from benign biliary tract obstructive diseases.

Management of patients with hepatitis C infection and renal disease

Hepatitis C virus(HCV) infection in patients with end-stage renal disease(ESRD) is associated with more rapid liver disease progression and reduced renal graft and patients' survival following kidney transplantation. Evaluations and management of HCV in patients with renal disease are challenging. The pharmacokinetics of interferons(IFN), ribavirin(RBV) and some direct acting antiviral(DAA), such as sofosbuvir, are altered in patients with ESRD. With dose adjustment and careful monitoring, treatment of HCV in patients with ESRD can be associated with sustained virological response(SVR) rates nearly comparable to that of patients with normal renal function. DAA-based regimens, especially the IFNfree and RBV-free regimens, are theoretically preferred for patients with ESRD and KT in order to increase SVR rates and to reduce treatment side effects. However, based on the data for pharmacokinetics, dosing safety and efficacy of DAA for patients with severe renal impairment are lacking. This review will be focused on the evaluations, available pharmacologic data, and management of HCV in patients with severe renal impairment, patients who underwent KT, and those who suffered from HCV-related renal disease, according to the available treatment options, including DAA.

Risk factors for endophthalmitis after cataract surgery in diabetic patients: a case control study

AIM: To identify risk factors associated with post-cataract surgery endophthalmitis(PCE) in type 2 diabetic patients.METHODS: A hospital-based retrospective case-control study was conducted on 194 type 2 diabetic patients undergoing cataract surgery in Rajavithi Hospital from January 2007 to December 2015. Fifteen patients with PCE were included as the case group and 179 patients without PCE were included as the control group. Potential factors associated with PCE among both groups including demographics, pre-operative characteristics, surgical settings and complications, were statistically analyzed using Chi-square testing and a logistic regression model.RESULTS: Within the case group, 53% were females and the median age was 68 y. Univariate analysis of pre-operative characteristics, surgical settings and complications revealed that recent pre-operative fasting plasma glucose, insulin therapy, presence of diabetic retinopathy, and severe non-proliferative or proliferative diabetic retinopathy were significantly associated with PCE. In a multivariate analysis adjusting for blood glucose level, insulin treatment was the only significant factor associated with an increased risk of PCE(OR 3.9, 95%CI 1.0-15.0, P=0.04) compared to patients without insulin treatment. The most common causative organisms were gram-positive bacteria(89%). Staphylococcus species represented the most common group(67%). Median best corrected visual acuity at 1-month and 3-month follow-up was equal at 0.7 logMAR(20/100).CONCLUSION: The authors identify insulin treatment as the only risk factor associated with endophthalmitis after cataract surgery in type 2 diabetic patients. Further studies with serum levels of pre-operative glycated hemoglobin(HbA_(1c)) and post-operative fasting plasma glucose level are essential to truly demonstrate the role of peri-operative glycemic markers as a risk factor for PCE.

Bacterial infections other than spontaneous bacterial peritonitis in cirrhosis

Cirrhotic patients are immunocompromised with a high risk of infection.Proinflammatory cytokines and hemodynamic circulation derangement further facilitate the development of serious consequences of infections.Other than spontaneous bacterial peritonitis,bacteremia and bacterial infections of other organ systems are frequently observed.Gram-negative enteric bacteria are the most common causative organism.Other bacterial infections,such as enterococci,Vibrio spp.,Aeromonas spp.,Clostridium spp.,Listeria monocytogenes,Plesiomonas shigelloides and Mycobacterium tuberculosis are more prevalent and more virulent.Generally,intravenous third generation cephalosporins are recommended as empirical antibiotic therapy.Increased incidences of gram-positive and drug-resistant organisms have been reported,particularly in hospitalacquired infections and in patients receiving quinolones prophylaxis.This review focuses upon epidemiology,microbiology,clinical features and treatment of infections in cirrhosis other than spontaneous bacterial peritonitis,including pathogen-specific and liver diseasespecific issues.

Artemisinin resistance or tolerance in human malaria patients

Malaria is a major cause of morbidity and mortality in the developing world.This situation is mainly due to emergence of resistance to most antimalarial drugs currently available. Artemisinin-based combination treatments are now first-line drugs for Plasmodium falciparum (P.falciparum) malaria.Artemisinin(qinghaosu) and its derivatives are the most rapid acting and efficacious antimalarial drugs.This review highlights most recent investigations into the emergence of artemisinin resistance in falciparum malaria patients on the Thai-Cambodian border,a historical epicenter for multidrug resistance spread spanning over 50 years.The study presents the first evidence that highlights the parasites reduced susceptibility to artemisinin treatment by prolonged parasite-clearance times,raising considerable concern on resistance development.Although the exact mechanism of action remains unresolved,development of resistance was proposed based from both in vitro experiments and human patients.Lines of evidence suggested that the parasites in the patients are in dormant forms,presumably tolerate to the drug pressure.The World Health Organization has launched for prevention and/or containment of the artemisinin-resistant malaria parasites.Taken together,the emergence of artemisinin resistance to the most potent antidote for falciparum malaria,poses a serious threat to global malaria control and prompts renewed efforts for urgent development of new antimalarial weapons.

Antimalarial qinghaosu/artemisinin: The therapy worthy of a Nobel Prize

Malaria is a major cause of human morbidity and mortality in the tropical endemic countries worldwide. This is largely due to the emergence and spread of resistance to most antimalarial drugs currently available. Based on the World Health Organization recommendation, artemisinin-based combination therapies are now used as first-line treatment for Plasmodium falciparum malaria. Artemisinin or qinghaosu(Chinese name) and its derivatives are highly potent, rapidly acting antimalarial drugs. Artemisinin was discovered in 1971 by a Chinese medical scientist Youyou Tu, who was awarded the Nobel Prize in 2015 on her discovering the antimalarial properties of qinghaosu from the traditional Chinese qinghao plant. Nevertheless, artemisinin resistance in falciparum malaria patients has first emerged on the Thai-Cambodian border in 2009, which is now prevalent across mainland Southeast Asia from Vietnam to Myanmar. Here, we reviewed malaria disease severity, history of artemisinin discovery, chemical structure, mechanism of drug action, artemisinin-based combination therapies, emergence and spread of drug resistance, including the recent findings on mechanism of resistance in the falciparum malaria parasite. This poses a serious threat to global malaria control and prompts renewed efforts for the urgent development of new antimalarial drugs.

Putative metabolic roles of the mitochondria in asexual blood stages and gametocytes of Plasmodium falciparum

Upon infection into human red cell,Plasmodium falciparum differentiates into asexual and sexual(gametocyte) stages.The mitochondrion is a tubular-cristate organelle,functionally and structurally different between the two stages.Genes and proteins involving metabolic and functional roles,protein targeting and import to this organelle, are comprehensively reviewed.The genes and proteins of the electron transport system are identified, partially characterized in human and rodent malaria parasites consisting of a single subunit of NADH dehydrogenase, two subunits of succinate dehydrogenase,cytochrome C reductase and cytochrome Coxidase.One of the primary functional roles of the mitochondrion in the parasite is the coordination of pyrimidine biosynthesis, the electron transport system and oxygen utilization through dihydroorotate dehydrogenase.All enzymes of tricarboxylic acid cycle,pyruvate dehydrogenase complex and some enzymes of ATP synthase,are identified and partially characterized using the completed P.falciparum genome.Some metabolic and functional roles of the organelle include oxidative phosphorylation,ubiquinone and heme biosynthesis,antioxidant defense and redox balance.Recent physiological studies involve membrane potential maintenance,cellular signaling and cation homeostasis.The organelle is a target for antimalarial drug,i.e.atovaquone.Based on the lines of evidence, we hypothesize that the parasite exhibits metabolic adaptation of the underdeveloped mitochondrial organelle to life in the mosquito vector and the human host.

Asymptomatic malaria infections among foreign migrant workers in Thailand

Objective:To determine the prevalence of malaria infections among foreign migrant workers in Thailand.Methods:Giemsa-stained thin and thick blood films were prepared from blood samples of 294 foreign migrant workers recruited in the study.Microscopic examination of these blood films was performed for malaria detection.Results:Blood film examination revealed 1.36%malaria infections in these 294 subjects.All positive cases were male Myanmar workers in which their blood films only ring stage of Plasmodium spp.was found at low parasite density (mean= 144 parasites/μL of blood).The prevalence of malaria infections was not significantly different among foreign migrant workers classified by age.gender,and resident province(P】0.05). Thin blood films of these workers also showed 78.91%hypochromic erythrocytes and 61.9% relative Eosinophilia.Conclusions:These findings indicate a high risk of malaria transmission. Therefore aclive malaria surveillance by using molecular methods with more sensitive and specific than microscopy should be considered for malaria control in foreign migrant workers.

Lead levels of Culex mosquito larvae inhabiting lead utilizing factory

Objective:To determine lead level primarily in Culex quinquefasciatus(Cx.quinquefasciatus), and Culex gelidus(Cx.gelidus) larvae inhabiting lead consuming factories,and to putatively estimate eco-toxicological impact of effluents from the firms.Methods:Third instars larvae were sampled by standard dipping method and lead concentrations in the larvae and their respective surrounding factory aquatic environments were determined through standard atomic absorption spectrophotometry(AAS).Results:Cx.quinquefasciatus was the most abundant species followed by Cx.gelidus.The levels of lead were higher in the Cx.quinquefasciatus(1.08-47.47μg/g),than in the wastewaters surface(0.01-0.78 μg/mL) from the factories or closer areas around factories. Other species were not reaching the.criteria for lead determination.Conclusions:The Cx. quinquefasciatus larvae can bio-accumulate the metal and can potentially serve as a biomarker of lead contamination,to complemente conventional techniques.

Circulating hTERT mRNA as a tumor marker in cholangiocarcinoma patients

AIM： To investigate human telomerase reverse transcriptase （hTERT） mRNA in the serum of cholangiocarcinoma patients. METHODS： The serum of thirty three cholangiocarcinoma patients, forty one benign biliary tract disease patients and ten healthy volunteers were collected and analyzed for the expression of hTERT mRNA by real-time reverse transcriptase-polymerase chain reaction （RT-PCR）. We then examined the correlation between values of serum hTERT mRNA and the pathological staging of cholangiocarcinoma. RESULTS： hTERT mRNA was detected in 28 of 33 （84.85%） of serum obtained from cholangiocarcinoma patients and 9 of 41 （21.9%） of serum obtained from benign biliary tract disease patients, hTERT mRNA was not detected in any serum obtained from healthy volunteers, on the other hand the common tumor marker, CA19-9 was detected in 20 of 33 （60.6%） of serum obtained from cholangiocarcinoma patients and 8 of 41 （19.5%） of serum obtained from benign biliary tract disease patients. However, no correlation was found between the present of serum hTERT mRNA and tumor staging. CONCLUSION： These results indicate that the detection of circulating hTERT mRNA was identified in almost all cholangiocarcinoma patients. It offers a novel tumor marker, which can be used as a complementary study for diagnosis of cholangiocarcinoma .

Effects of α-mangostin on apoptosis induction of human colon cancer

AIM:To investigate the effect of α-mangostin on the growth and apoptosis induction of human colon cancer cells.METHODS:The three colorectal adenocarcinoma cell lines tested (COLO 205,MIP-101 and SW 620) were treated with α-mangostin to determine the effect on cell proliferation by MTT assay,cell morphology,chromatin condensation,cell cycle analysis,DNA fragmentation,phosphatidylserine exposure and changing of mitochondrial membrane potential.The molecular mechanisms of α-mangostin mediated apoptosis were further investigated by Western blotting analysis including activation of caspase cascade,cytochrome c release,Bax,Bid,p53 and Bcl-2 modifying factor.RESULTS:The highest inhibitory effect of α-mangostin on cell proliferation of COLO 205,MIP-101 and SW 620 were 9.74 ± 0.85 μg/mL,11.35 ± 1.12 μg/mL and 19.6 ± 1.53 μg/mL,respectively.Further study showed that α-mangostin induced apoptotic cell death in COLO 205 cells as indicated by membrane blebbing,chromatin condensation,DNA fragmentation,cell cycle analysis,sub-G1 peak (P < 0.05) and phosphatidylserine exposure.The executioner caspase,caspase-3,the initiator caspase,caspase-8,and caspase-9 were expressed upon treatment with α-mangostin.Further studies of apoptotic proteins were determined by Western blotting analysis showing increased mitochondrial cytochrome c release,Bax,p53 and Bmf as well as reduced mitochondrial membrane potential (P < 0.05).In addition,up-regulation of tBid and Fas were evident upon treatment with α-mangostin (P < 0.01).CONCLUSION:α-Mangostin may be effective as an anti-cancer agent that induced apoptotic cell death in COLO 205 via a link between extrinsic and intrinsic pathways.

Prognostic relevance of circulating CK19 mRNA in advanced malignant biliary tract diseases

AIM: To determine the role of circulating tumor cells (CTCs) in prediction of the overall survival of patients with advanced malignant biliary tract obstruction. METHODS: We investigated the prognostic value of CTCs by examining two markers, cytokeratin (CK) 19 and human telomerase reverse transcriptase (hTERT) mRNA, in 40 patients diagnosed with advanced malig- nant biliary tract diseases. Quantitative real-time re- verse transcription polymerase chain reaction was used to detect CK19 and hTERT mRNA in the peripheral blood of these patients. Overall survival was analyzed using the Kaplan-Meier method and Cox regression modeling.RESULTS: Positive CK19 and hTERT mRNA expression was detected in 45% and 60%, respectively, of the 40 patients. Univariable analysis indicated that positive CK19 mRNA expression was significantly associated with worse overall survival (P = 0.009). Multivariable analysis determined that positive CK19 mRNA expres- sion, patient's age and serum bilirubin were each inde- pendently associated with overall survival. CONCLUSION: CK19 mRNA expression levels in pe- ripheral blood appear to provide a valuable marker to predict the overall survival of patients with advanced malignant biliary tract obstruction.

Anticancer activity of Δ^(9)-tetrahydrocannabinol and cannabinol in vitro and in human lung cancer xenograft

Objective:To investigate the effects of Δ^(9)-tetrahydrocannabinol,the principal psychoactive compound of Cannabis sativa,and cannabinol,a Δ^(9)-tetrahydrocannabinol degradative product,on human non-small cell lung cancer cells.Methods:Δ^(9)-Tetrahydrocannabinol and cannabinol were tested for anticancer activity in human non-small cell lung cancer(A549)cells.The effects on cell proliferation,apoptosis,and phosphorylation profiles were examined.The effects of Δ^(9)-tetrahydrocannabinol and cannabinol on tumor growth were also investigated using a xenograft nude mouse model.Apoptosis and targeted phosphorylation were verified by immunohistochemistry.Results:Δ^(9)-Tetrahydrocannabinol and cannabinol significantly inhibited cell proliferation and increased the number of apoptotic cells in a concentration-dependent manner.The Δ^(9)-tetrahydrocannabinol-and cannabinol-treated cells had lower levels of phosphorylated protein kinase B[AKT(S473)],glycogen synthase kinase 3 alpha/beta,and endothelial nitric oxide synthase compared to the controls.The study of xenograft mice revealed that tumors treated with 15 mg/kg Δ^(9)-tetrahydrocannabinol or 40 mg/kg cannabinol were significantly smaller than those of the control mice.The tumor progression rates in mice treated with 15 mg/kg Δ^(9)-tetrahydrocannabinol or 40 mg/kg cannabinol were significantly slower than in the control group.Conclusions:These findings indicate that Δ^(9)-tetrahydrocannabinol and cannabinol inhibit lung cancer cell growth by inhibiting AKT and its signaling pathways,which include glycogen synthase kinase 3 alpha/beta and endothelial nitric oxide synthase.