Effect of insulin-sensitizing agents in combination with ezetimibe, and valsartan in rats with non-alcoholic fatty liver disease

AIM： To assess whether treatment with insulinsensitizing agents （ISAs） in combination with ezetimibe and valsartan have greater effect on hepatic fat content and lipid peroxidation compared to monotherapy in the methionine choline-deficient diet （MCDD） rat model of non-alcoholic fatty liver disease （NAFLD）. METHODS： Rats （n = 6 per group） were treated with different drugs, including MCDD only, MCDD diet with either metformin （200 mg/kg）, rosiglitazone （3 mg/kg）, metformin plus rosiglitazone （M＋R）, ezetimibe （2 mg/ kg）, valsartan （2 mg/kg）, or combination of all drugs for a total of 15 wk. Liver histology, lipids, parameters of oxidative stress and TNF-alpha were measured. RESULTS： Fatty liver （FL） rats demonstrated severe hepatic fatty infiltration （〉 91% fat）, with an increase in hepatic TG （＋1263%, P 〈 0.001）, hepatic cholesterol （＋245%, P 〈 0.03）, hepatic MDA levels （＋225%, P 〈 0.001）, serum TNF-alpha （17.8 ＋ 10 vs 7.8 ＋ 0.0, P 〈 0.001）, but a decrease in hepatic alpha tocopherol （-74%, P 〈 0.001） as compared to the control rats. Combination therapy with all drugs produced a significant decrease in liver steatosis （-54%）, hepatic TG （-64%）, hepatic cholesterol （-31%） and hepatic MDA （-70%）, but increased hepatic alpha tocopherol （＋443%） as compared to FL rats. Combination therapy with ISA alone produced a smaller decrease in liver steatosis （-32% vs -54%, P 〈 0.001） and in hepatic MDA levels （-55% vs -70%, P 〈 0.01）, but a similar decrease in hepatic lipids when compared with the all drugs combination. TNF-alpha levels decreased significantly in all treatment groups except in ISA group. CONCLUSION： Combination therapies have a greater effect on liver fat content as compared to monotherapy. Rosiglitazone appears to improve hepatic steatosis to a greater extent than metformin.

Fenofibrate for patients with asymptomatic primary biliary cirrhosis

AIM:Primary biliary cirrhosis (PBC) is a chronic, cholestatic disease of autoimmune etiology,the histology of which shows a destruction of the intrahepatic bile duct and portal inflammation. Ursodeoxycholic acid (UDCA) is now used as a first-line drug for asymptomatic PBC (aPBC) because it is reported that UDCA decreases mortality and prolongs the time of liver transplantation.However, only 20-30% of patients respond fully to UDCA.Recently,lipoprotein-lowering agents have been found to be effective for PBC.The aim of this study was to examine the safety and efficacy of fenofibrate, a member of the fibrate class of hypolipidemic and anti-inflammatory agent via peroxysome proliferatory-activated receptor α,in patients with aPBC.METHODS:Fenofibrate was administered for twelve weeks in nine patients with aPBC who failed to respond to UDCA.UDCA was used along with fenofibrate during the study.The data from aPBC patients were analyzed to assess the biochemical effect of fenofibrate during the study.RESULTS: The serum levels of alkaline phosphatase (ALP)(285±114.8IU/L) and immunoglobulin M (IgM) (255.8±85.9mg/dl) significantly decreased to 186.9±76.2IU/L and 192.9±67.5mg/dL respectively, after fenofibrate treatment in patients with aPBC (P<0.05). Moreover,the titer of antimitochondrial antibody (AMA) also decreased in 4 of 9 patients with aPBC. No adverse reactions were observed in any patients.CONCLUSION:Fenofibrate appears to be significantly effective in treating patients with aPBC who respond incompletely to UDCA alone.Although the mechanism of fenofibrate on aPBC has not yet been fully clarified,combination therapy using fenofibrate and UDCA might be related to the anti-immunological effects, such as the suppression of AMA production as well as its antiinflammatory effect.

Biochemical response to lamivudine treatment in HBeAg negative chronic hepatitis B patients in Iran

AIM： To study the effect of a one-year lamivudine regimen in patients with chronic hepatitis B. METHODS： Medical records of HBeAg negative hepatitis B patients who attended a hepatitis clinic in Tehran between March 2002-March 2004 were evaluated. The patients received 100 mg lamivudine tablets once daily for at least 12 mo. Liver enzymes and complete blood count were checked at baseline and the end of treatment （12th mo） and 6 mo after discontinuation of treatment. RESULTS： Of all patients, 24 were excluded. Of 71 patients left, 58 （81.7%） were men. Mean age of the patients was 38 ± 14 years. Mean level of ALT in serum was 1437 ± 205 nkat/L at baseline with a significant reduction at the end of treatment to a mean level of 723 ± 92 nkat/L （P = 0.002）. In 38 patients （53.5%）, the ALT level was normal after one-year treatment. Five patients （7.3%） relapsed （biochemically） within 6 mo after discontinuing lamivudine therapy （the patients with good end of treatment response）. Mean level of AST in serum was 1060 ± 105 nkat/L at baseline which decreased significantly to 652 ± 75 nkat/L at the end of treatment （P = 0.002）. CONCLUSION： Over half （53.5%） of chronic hepatitis B patients with HBeAg negative have normal liver enzyme level at 12-mo lamivudine therapy.

Nuclear factor-kappaB activation on the reactive oxygen species in acute necrotizing pancreatitic rats

AIM： To investigate the potential role of nuclear factor kappa-B （NF-κB） activation on the reactive oxygen species in rat acute necrotizing pancreatitis （ANP） and to assess the effect of pyrrolidine dithiocarbamate （PDTC, an inhibitor of NF-κB）.METHODS： Rat ANP model was established by retrograde injection of 5% sodium taurocholate into biliopancreatic duct. Rats were randomly assigned to three groups （10 rats each）： Control group, ANP group and PDTC group. At the 6^th of the model, the changes of the serum amylase,nitric oxide （NO）, malondialdehyde （MDA）, superoxide dismutase （SOD） and pancreatic morphological damage were observed. The expressions of inducible nitric oxide （iNOS） were observed by SP immunohistochemistry. And bhe expressions of NF-κB p65 subunit mRNA were observed by hybridization in situ.RESULTS： Serum amylase and NO level decreased significantly in ANP group as compared with PDTC administrated group [（7 170.40＋1 308.63） U/L vs（4 074.10＋1 719.78） U/L,P〈0.05], [（76.95±9.04） μmol/L vs （65.18±9.02） μmol/L,P〈0.05] respectively. MDA in both ANP and PDTC group rose significantly over that in control group [（9.88＋1.52）nmol/L, （8.60±1.41） nmol/L, vs （6.04：hl.78） nmol/L,P〈0.05], while there was no significant difference between them. SOD levels in both ANP and PDTC group underwent a significant decrease as compared with that in control[（3 214.59±297.74） NU/mL, （3 260.62±229.44） NU/mL,vs（3 977.80＋309.09） NU/mL, P〈0.05], but there was no significant difference between them. Though they were still higher bhan those in Control group, pancreas destruction was slighter in PDTC group, iNOS expression and NF-κB p65 subunit mRNA expression were lower in PDTC group as compared with ANP group.CONCLUSION： We conclude that correlation among NF-κB activation, serum amylase, reactive oxygen species level and tissue damage suggests a key role of NF-κB in the pathogenesis of ANP. Inhibition of NF-κB activation may reverse the pancreatic damage of rat ANP and the production of reactive oxygen species.

Phase III study of TAC and TP regimens as neoadjuvant chemotherapy in patients withtriple-negative breast cancer

Objective： This study aimed to compare the efficacy and safety of neoadjuvant chemotherapy with TAC and TP regimens of triple negative breast cancer （TNBC）. Methods： A total of 102 patients with TNBC were confirmed by histopathology. They were divided into TAC group （52 cases） and TP group （50 cases）. Group TAC： Docetaxel 75 mg/m2 or paclitaxel （taxol liposome） 135 mg/m2 on all, pirarubicin 40 mg/m2 or epirubicin 75 mg/m2 on d2, cyclophosphamide 600 mg/m2 on dl; Group TP： Docetaxe175 mg/m2 or paclitaxel （taxol liposome） 135 mg/m2 on dl, cisplatin 30 mg/m2 on d2-d4, with 21 days as a cycle. All patients underwent operation after 2-4 cycles of chemotherapy. The short-term effects and toxic and adverse effects were evaluated. Results： In TAC group, 5 cases （9.6%） had pathological complete release （pCR）, 35 cases （67.3%） partial release （PR）, 9 cases （17.3%） stable disease （SD）, and the response rate （RR） was 76.9%. In TP group, 4 cases （8%） had pCR, 32 cases （64%） PR, 5 cases （10%） SD, and RR was 72%. In 102 patients, 12 patients with tumor progression after 2 cycles of chemotherapy, included 3 cases in TAC group, 9 cases in TP group. In TAC group, 2 cases occurred atrial premature contraction; while 3 cases developed grade 2 renal injury in TP group. In TAC group, grade 3-4 hematologic toxicity and alopecia was significantly higher than that in TP group, but grade 3-4 gastrointestinal reaction rate in TP group was significantly higher than TAC group. Conclusion： TAC and TP regimens all had certain efficacy in the neoadjuvant chemotherapy for TNBC, and the toxicity reactions can be tolerated.

Clinical Application of Multiplex PCR Assay for the Diagnosis of the Etiology of Genital Ulcer Disease Among Patients Attending STD Clinics in Guangzhou, China

Objectives: To develop a method of simultaneous PCR detection of Haemophilus ducreyi, Treponema pallidum, andHerpes Simplex Virus Types 1 and 2 from genital ulcersamong patients attending STD clinics in Guangzhou, China;and evaluate the clinical application of multiplex PCR (M-PCR) assay for diagnosing the etiology of genital ulcerdiseases (GUD). Methods: 244 patients with a genital ulcer were evaluated.Clinical etiology of GUD was based on physical appearanceand microbiologic evaluations that included darkfieldmicroscopy examination (D-F) and serology test for syphilis(STS). Swabs of each genital ulcer were tested for HSVantigen by enzyme immunoassay (EIA) and processed in anM-PCR assay for simultaneous detection of T pallidum, HSVand H ducreyi. Results: The standard strains of T pallidum, HSV and Hducreyi were amplified by M-PCR, producing amplifiedproducts of 260bp,432bp,170bp, respectively. The sensitivityof M-PCR is 10~2 pg DNA. M-PCR assay for T.pallidum, HSVand H ducreyi showed good agreement en compared with D-F detection for T pallidum, STS, H ducreyi culture and EIAfor HSV antigen (Kappa scores are 0.774,0.704,0.793,0.756,respectively). Conclusions: The M-PCR is a convenient, accurate andreliable assay for the detection of T pallidum, HSV and Hducreyi from genital ulcers, and can be used as a method ofdiagnosing the etiology of GUD.

Effects of phospholipase D on cardiopulmonary bypass-induced neutrophil priming

Objective:To investigate the relationship betwe en phospholipase D (PLD) activation and neutrophil priming induced by cardiopulm onary bypass (CPB), and try to clarify whether CPB-induced systemic inflammator y response can be attenuated by inhibiting neutrophilic PLD activation. Methods:Neutrophils were isolated from arterial blood of 8 pat ients undergoing valve replacement before operation and 30 min after initiation of CPB respectively. Both the preoperative and CPB-stirred neutrophils were sub divided into 5 groups by receiving different experimental interventions: (1) bac terial lipopolysaccharide (LPS, 10 ng·ml -1 ), (2) N-formylmethionylph enylalanine (fMLP, 1 μmol·L -1 ), (3) LPS+fMLP, (4) 1-butanol ( 0.5 %)+ LPS+fMLP, (5) vehicle. Elastase and myeloperoxidase (MPO) release was measured f or the parameters of neutrophil activation, neutrophil PLD activity was determin ed by quantitation of choline produced from the stable product of phosphatidylch oline catalyzed by PLD. Results:(1) Preoperative neutrophils treated with LPS+fMLP pre sented significantly higher PLD activity ( 13.48 ± 2.61 nmol choline·h -1 ·mg -1 ) and released more elastase and MPO than cells treated with v ehicle (PLD activity 3.70 ± 0.49 nmol choline·h -1 ·mg -1 , P< 0.01 ), LPS (P< 0.01 ) and fMLP respectively. In 1-butanol+LPS+fMLP group, PLD activity of preoperative neutrop hils was lower than that in LPS+fMLP group (P< 0.01 ), b esides the release of elastase and MPO decreased sharply below both LPS + fMLP a nd fMLP groups (P< 0.01 ). In LPS group, PLD activity wa s higher (P< 0.01 ), while elastase and MPO release did not differ from control. fMLP group presented PLD activity, elastase and MPO rel ease higher than control (P< 0.01 ); nevertheless, lower than LPS+fMLP group (P< 0.01 ). (2) CPB-stirred neutro phils presented prominent PLD activity increment, and even the control level was 3.59 -fold of the pre-operative control (P< 0.01 ) . PLD activity in LPS+fMLP group was higher than that in other groups. Notably, PLD activity was even nonstatistically lower in 1-butanol+LPS+fMLP group than t hat in LPS or fMLP group. CPB-stirred neutrophils in LPS+fMLP group released mo re elastase and MPO than control, LPS, and 1-butanol+LPS+fMLP groups did ( P< 0.01 ); however, neither of the release was statistically different from that of fMLP group. Conclusions:Cardiopulmonary bypass enables neutrophil priming accompanied with significant increase in PLD activity. Inhibition of neutrophil PLD activation attenuates its priming and may alleviate CPB-induced systemic in flammatory reaction.