Objective To analyze the differential expression of microRNA-146a (miR-146a) in monocyte-macrophage cell line (THP-1 cells) after induction by Cryptococcus neoformans (C.neoformans,reference strain WM148) or Cryptococcus gattii (C.gattii,reference strain R265),and investigate the mechanism of miR-146a in regulating the inflammatory response of cryptococcal meningitis.Methods The cultured THP-1 cells were divided into two groups to be induced by C.neoformans or C.gattii,respectively.THP-1 cells were induced with inactivated WN148 (or R265) strains at multiplicity of infection (MOI) of 5 in all experiments.The supematant and the cell pellet were collected separately after incubation.The expression of miR-146a was measured by real-time quantitative PCR (qRT-PCR) technique.The levels of TNF-u and IL-6 release were assayed by ELISA.Results The expression of miR-146a increased significantly in the C.neoformans induction group compared to 0 h.It reached peak at 3 h (P＜0.01),and then declined gradually.The level of TNF-α increased in supematant and reached peak at 12 h.The expression of IL-6 did not change significantly at each time point.The expression of miR-146a and TNF-α increased gradually and reached peak at 12 h in the C.gattii induction group (P ＜0.01),but the change did not reach statistical significance at 3 h,6 h time points.The expression of IL-6 gradually increased,and reached peak at 12 h time point.Conclusions Following stimulation with C.neoformans or C.gattii,the expression ofmiR-146a in THP-1 cells showed different patterns over time.The expression levels of TNF-α and IL-6 showed different patterns.These findings suggest that there may be different regulatory mechanisms in the THP-1 cells-associated inflammatory response after stimulation by inactivated C.neoformans and C.gattii strains.

Objective To explore the treatments and their effects on painless chronic pancreatitis (CP) in adult. Methods A historical cohort study of adult subjects with CP, who were hospitalized in our center in the last decade, was performed, and the clinical data were analyzed. Results There were 56 cases of adult painless CP, and 46 (82.1%) (31 males and 15 females, age 51.6±14.5 years) were followed up(34.5±27.1 months). Imaging findings showed 26 cases of pancreatic duct stones and morphological changes. These patients received endoscopic intervention therapy(n=14), surgical procedures(n=7), combined endoscopic and surgical treatment(n=3) and conservative treatment(n=2). Another group of 18 cases of pancreatic or peripheral occupying lesions received surgery. During follow-up of the above 2 groups, 1 patient died of pancreatic cancer in each group. The symptoms of diabetes and diarrhea were not relieved, but there was improvement of general condition in some patients. Stones re-occurred after surgery in one patient. Two patients with pancreatic duct dilation and cysts underwent endoscopic treatment(n=1) and surgical procedure(n=1), respectively. But the latter patient experienced gastric bleeding, cirrhosis and jaundice with unknown reason after the surgery. Conclusion In adult patients with CP, endoscopic and surgical procedures had similar effect in patients with pancreatic duct stones and dilation. Surgical procedures are preferably performed in those with pancreatic occupying lesions. Further study is needed in those only with pancreatic duct dilation. The above treatments can improve general conditions in some patients. Intensive surveillance is suggested to the patients with painless CP.

Objective To investigate the changes of indications, degree of difficulty in procedure, complication and its severity in endoscopic retrograde cholangiopancreatography (ERCP) in Changhai hospital from 2001 to 2007. Methods The clinical data, including demographic data, indications, degree of difficulty in procedure, success rate, complication rate and severity of complication, of 2374 patients who underwent ERCP in 2001 and 2007 (966 in 2001 and 1408 in 2007), were retrospectively reviewed. Results Indications of ERCP changed at an interval of 5 years. Operations due to bile duct stone decreased (59.0% vs. 49.3%, P=0.000), while operations due to pancreas disease, especially chronic pancreatitis (6.6% vs. 18.5%, P=0.000) and recurrent pancreatitis (0.2% vs.1.6%, P=0.001), increased. Patients with biliary duct problems after liver transplantation appeared in 2007. The procedures of ERCP performed in 2007 were more difficult (P=0.000), with an increased percentage of Degree 5 procedure (7.3% vs. 33.3%, P=0.000). The number of diagnostic ERCP significantly decreased (Degree 1 + Degree 3, 30.5% +2.8% vs. 5.9% +3.1%, P=0.000). There was no significant difference in the success rate between the two years (P=0.084). The complication rate of ERCP in 2007 was significantly higher than that in 2001 (3.73% vs. 7.88%, P=0.000), but the severity of complication showed no significant difference (P=0.820). Conclusion Cases of diagnostic ERCP decreased in 2007. Indications of ERCP have changed, with a decrease in bile duct diseases and an increase in pancreatic diseases. The procedures are more complicated, but it does not lead to lower success rate. The increase in complication rate is possibly due to increase of therapeutic ERCP.

According to the current situations and development of (TCMIs), the author of the article reveals the scientific connotation of TCMIs in theory, preparations and clinic application, and points out that TCMIs are an innovative and breakthrough of conventional dosage forms of traditional Chinese medicines, the combination of traditional theory and modern technology as well as a type of modern dosage form with the characteristics of traditional Chinese medicines, which conforms to the principle of including the essence and excluding the wastes for traditional Chinese medicine preparations, meets the demands for quick-acting of traditional Chinese medicines and guides one of the development orientation of traditional Chinese medicines. In the meantime, an analysis was also made on key issues, such as adverse reactions of TCMIs, modern clinical application, special drug delivery route and diversity of components and ingredients.
Drug Delivery Systems ; methods ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Exanthema ; chemically induced ; Humans ; Injections ; adverse effects ; Medicine, Chinese Traditional ; adverse effects ; methods ; trends ; Nausea ; chemically induced ; Product Surveillance, Postmarketing ; methods ; statistics & numerical data ; Vomiting ; chemically induced

OBJECTIVETo elucidate the effect of neurotrophin p75 receptor (p75NTR)on transmural dispersion repolarization (TDR) of the layers of left ventricular myocytes in rabbits with myocardial infarction (MI).

METHODSForty Japanese rabbits were divided into four groups (n = 10): (1) Sham group, (2) Heald myocardial infarction (HMI) group, (3) p75 NTR activation group, (4) p75 NTR inhibition group. Cardiomyocytes were isolated with enzyme digestion and the currents were recorded by whole-cell patch-clamp technique.

RESULTSCompared with those in the sham group, the duration of 90% action potential repolarization (APD90) and transmural dispersion repolarization of three layers of left ventricular myocytes were obviously raised (P < 0.05). But significant reduction was observed in p75NTR(-) group. Current densities of Ito and I(Ks, tail) in the p75NTR(+) group and HMI group were significantly reduced (P < 0.05), especially in mid myocytes. And no obvious changes were observed in p75NTR(-) group.

CONCLUSIONActivation of p75NTR(+) increases transmural dispersion repolarization, which may lead to the incidence of arrhythmia.

Animals ; Arrhythmias, Cardiac ; physiopathology ; Heart Ventricles ; physiopathology ; Membrane Potentials ; Myocardial Infarction ; physiopathology ; Myocytes, Cardiac ; physiology ; Patch-Clamp Techniques ; Rabbits ; Receptor, Nerve Growth Factor ; metabolism

Background Choroidal melanoma (CM)is the most common primary intraocular tumor,and brachytherapy is one of the most common therapeutic modality in the treatment of the tumor.However,this irradiation approach has not been evaluated in China. Objective The present study was to analyze the effectiveness and safety of domestic 125I plaque irradiation in the treatment of CM. Methods Forty New Zealand albino rabbits were randomized into 5 groups with 8 rabbits 8 eyes (right eyes) in each group.CM models were established in 16 of 40 New Zealand albino rabbits by implanting the rat B16F10 melanoma cell fragments into the suprachoroidal space of right eyes.After 3 weeks,domestic 125I plaque was fixed at the location of CM in the irradiation group 1,and 8 rabbits with CM served as model control group.The clinical effectiveness of 125I plaque for CM was evaluated based on the fundus examination with indirect ophthalmoscopy,B scan ultrasonography,fundus photographs and color Dopplerimaging.Regarding the safety study,domestic 125I plaque was fixed on the normal right cycs of normal rabbits,while the plaques without 125I seeds were used as the sham group.No intervene was performed in the rabbits of blank group.The number of CD4+,CD8+ T cells in peripheral blood was detected by flow cytometry before plaques implanted and on 3,7,15 and 30 days after the plaque was removed.The animals were sacrificed and the eyes were obtained for histology examination.The use of the experimental animals complied with Statement of ARVO. Results After implantation of B16F10 melanoma cell fragments,CM grew steadily and rapidly with the similar size between irradiation group 1 and model control group ( P =0.550).One week after administration of the treatment,tumor size was(0.31±0.07 )cm in irradiation group 1 and (0.85±0.18 )em in the model control group,with the significant difference between them( P=0.001 ).Two week after application of 125I plaque,the size of tumor was smaller than that before irradiation (P=0.007 ).Histologically,the tumors were mostly limited beneath the pigment epithelial layer with less neovascularization,fibrosis in the tumor was found in some area in the irradiation group when compared with model control group.No significant differences were found in the proportions of CD4+,CD8+ T cells and CD4+/CD8+ at different time points in the irradiation groups of normal eyes and sham group (Fgroup =0.770,8.110,2.230; P=0.380,0.060,0.140; Ftime =0.770,3.220,4.230; P =0.550,0.170,0.004 ).Chronic inflammatory cells infiltration cornea,subconjunctival epithelial and selera surface,but sclera had no necrosis and organization.Conclusions These results suggest that domestic 125I plaque irradiation is effective for the treatment of CM,and has limited side effects on normal rabbits.

This study was aimed to investigate the effect of stress induced by high-intensity exercises on the cardiovascular system. In the epidemiological investigation, 200 subjects (test group) engaged in special high-intensity exercises, and 97 who lived and worked in the same environment and conditions as those in the test group, but did not participate in the exercises served as controls. In the second part of the study, 50 mice were randomly divided into control group, exhaustive swimming group, white noise group, exhaustive swimming plus white noise group, and pioglitazone intervention group. The results showed that the plasma concentrations of the myocardial injury markers heart fatty acid-binding protein (H-FABP), C-reactive protein (CRP), β-endorphin (β-EP) and levels of psychological stress were significantly increased in test group as compared with control group; special high-intensity exercises resulted in a significant elevation of the incidence of cardiac arrhythmias. Animal experiments showed that the plasma levels of corticosterone (CORT) and troponin I (TnI) were raised while the level of SOD was reduced in exhaustive swimming group, white noise group, and exhaustive swimming plus white noise group. The expression levels of PPARγ mRNA and protein were decreased in myocardial tissues in these groups as well. HE staining showed no remarkable change in myocardial tissues in all the groups. Treatment with pioglitazone significantly decreased the plasma levels of TnI and CORT, while increased the level of SOD and the expression levels of PPARγ mRNA and protein. It was concluded that the high-intensity exercises may induce a heavy physical and psychological stress and predispose the subjects to accumulated fatigue and sleep deprivation; high-intensity exercises also increases the incidence of arrhythmias and myocardial injury. PPARγ may be involved in the physical and psychological changes induced by high-intensity exercises.
Animals ; Heart Injuries ; physiopathology ; Male ; Mice ; PPAR gamma ; genetics ; physiology ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Stress, Physiological

This article reports the investigation of the effect of carvedilol (Car) on T-type calcium current (I(Ca,T)) of noninfarcted ventricular myocytes in rabbit models of healed myocardial infarction (HMI). Rabbits with left anterior descending artery ligation were prepared and allowed to recover for 8 weeks, as HMI group. Animals undergoing an identical surgical procedure without coronary ligation were served as the sham-operated group (sham group). Whole cell voltage-clamp techniques were used to measure and compare currents in cells from the different groups. Noting that I(Ca,T) density in HMI cells increased markedly to -2.36 +/- 0.12 pA/pF (at -30 mV) compared with cells of sham, where little I(Ca,T) (-0.35 +/- 0.02 pA/pF) was observed. Meanwhile, further analysis revealed a significant hyperpolarizing shift of steady-state activation curve of I(Ca,T) in HMI cells, where the time constants of deactivation were prolonged and the time of recovery from inactivation was shortened. Finally, the amplitude of I(Ca,T) was increased. Carvedilol (1 micromol x L(-1)) was found to decrease the amplitude of I(Ca,T) to -1.38 +/- 0.07 pA/pF through inhibiting process of I(Ca,T) activation. Furthermore, carvedilol delayed recovery from inactivation of I(Ca,T) and shortened the time constants of deactivation in HMI cells. This study suggested that the application of carvedilol in HMI cells contributes to the dynamic changes in I(Ca,T) and may account for reduction of incidence of arrhythmia after myocardial infarction.
Adrenergic beta-Antagonists ; pharmacology ; Animals ; Calcium Channels, T-Type ; drug effects ; Carbazoles ; pharmacology ; Female ; Male ; Myocardial Infarction ; pathology ; physiopathology ; Myocytes, Cardiac ; drug effects ; physiology ; Patch-Clamp Techniques ; Propanolamines ; pharmacology ; Rabbits

BACKGROUNDThe rapidly activating delayed rectifier potassium current (I(Kr)), whose pore-forming alpha subunit is encoded by the human ether-a-go-go-related gene (hERG), is a key contributor to the third phase of action potential repolarization. The aim of this study was to investigate the effect and mechanism of arecoline hydrobromide induced inhibition of hERG K(+) current (I(hERG)).

METHODSTransient transfection of hERG channel cDNA plasmid pcDNA3.1 into the cultured HEK293 cells was performed using Lipofectamine. A standard whole-cell patch-clamp technique was used to record the I(hERG) before and after the exposure to arecoline.

RESULTSArecoline decreased the amplitude and the density of the I(hERG) in a concentration-dependent manner (IC(50) = 9.55 mmol/L). At test potential of +60 mV, the magnitude of I(hERG) tail at test pulse of -40 mV was reduced from (151.7 ± 6.2) pA/pF to (84.4 ± 7.6) pA/pF (P < 0.01, n = 20) and the magnitude of I(hERG) tail at test pulse of -110 mV was reduced from (-187.5 ± 9.8) pA/pF to (-97.6 ± 12.6) pA/pF (P < 0.01, n = 20). The blockade of arecoline in the open and inactivated state was significant in a state-dependent manner. The maximal blockade was achieved in the inactivated state. Studies of gating mechanism showed that the steady-state activation curve of I(hERG) was significantly negatively shifted by arecoline. Time constants of activation were shortened. Steady-state inactivation curve and time constants of fast inactivation were not significantly affected by arecoline. Furthermore, the inhibition of I(hERG) by arecoline was characterized markedly by a frequency-dependent manner from 0.03 to 1.00 Hz pulse.

CONCLUSIONArecoline could potently block I(hERG) in both frequency and state-dependent manner.

Action Potentials ; drug effects ; Arecoline ; pharmacology ; Dose-Response Relationship, Drug ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; antagonists & inhibitors ; physiology ; HEK293 Cells ; Humans