[Alteration on hepatocyte nuclear factor 1α expressions and significance in the process of occurrence and development of liver inflammation and fibrosis in patients with chronic hepatitis B].

Objective: To investigate the relationship between the expression of hepatocyte nuclear factor-1 α (HNF-1α) and the occurrence and development of liver inflammation and fibrosis in liver tissues of patients with chronic hepatitis B. Methods: Sixty-four patients with chronic hepatitis B who were diagnosed and treated in our hospital from 2011 to 2018 were selected. All patients underwent ultrasound-guided aspiration liver biopsy. The pathological results of liver biopsy were collected for inflammation grading and fibrosis staging. The liver puncture biopsies was collected by paraffin sectioning. The expression of HNF1α in the liver tissue was detected by immunohistochemical staining. Mantel-Haenszel χ(2) test was used for bidirectional ordered grouping data, and Spearman's rank-correlation test was used for rank correlation analysis. Results: There were varying degrees of inflammatory necrosis and fibrosis in the liver tissues of patients with chronic hepatitis B. There was a linear relationship between the expression of HNF1α and the level of inflammation in liver tissues (χ (2)(MH) = 40.70, P < 0.05). The expression of HNF1α in liver tissues of patients with chronic hepatitis B was decreased with the increase of liver inflammation. The expression intensity of HNF1α was negatively correlated with the inflammation grade (r(s) = -0.815, P < 0.05). There was a linear relationship between the expressions of HNF1α and the degree and stage of liver fibrosis (χ (2)(MH) = 31.95, P < 0.05). The expression level of HNF1α in liver tissue was gradually decreased with the aggravation of liver fibrosis. The expression intensity of HNF1α was negatively correlated with fibrosis stage (r(s) = -0.713, P < 0.05). Conclusion: HNF1α is closely related to the occurrence and development of liver tissue inflammation and fibrosis, and is expected to be a sensitive indicator for evaluating the level of liver tissue inflammation and fibrosis in patients with chronic hepatitis B. In addition, its down-regulation may be involved in the process of occurrence and development of liver inflammation and liver fibrosis, and may become a new target for the treatment of chronic hepatitis B.

Protective effect of sonic hedgehog signaling pathway activation on acute myocardial infarction.

To study changes in the sonic hedgehog (Shh) signaling pathway in acute myocardial infarction (AMI) and the protective effect of changes in Shh signaling pathway activity on AMI, specific pathogen-free (SPF) C57BL/6 mice were treated with left anterior descending (LAD) ligation to establish an AMI model. The samples were collected on the 1st, 3rd, 14th, and 21st days after AMI induction. After the operations, the mice were administered the Shh signaling pathway receptor agonist SAG1.3 (5 mg/kg/d) and antagonist SANT-1 (3.3 mg/kg/d) by intraperitoneal injection. The myocardial ischemia model was established by oxygen glucose deprivation (OGD) in vitro. The AMI mouse model and the in vitro OGD-induced myocardial ischemia model were established. The Smo agonist SAG1.3 was used to activate the Shh signaling pathway, thereby reducing the expression of Bcl-2 and Bax. The number of apoptotic cells was reduced. Administration of the antagonist SANT-1 inhibited Shh signaling pathway activity by increasing the expression of Bcl-2 and Bax, and the number of apoptotic cells increased. In conclusion, activation of the Shh signaling pathway improved cardiac functions and myocardial remodeling and reduced the apoptosis of myocardial cells.

[The role of lysosomes in manganese-induced toxicity in SK-N-SH cells].

Objective: To investigate the role of lysosomes in manganese-induced toxicity in human neuroblastoma SK-N-SH cells. Methods: SK-N-SH cells were treated with MnCl(2) at doses of 0.062 5, 0.125, 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L for 24 h, and the cell viability was detected by MTT assay. Cells were treated with MnCl(2) at doses of 0.125, 0.25, 0.5 and 1.0mmol/L for 24 h, and lysosomes labeled with lysotracker red were observed by laser confocal microscopy, the expression levels of LAMP1 and CTSD were detected by western blot, and CTSD activity was detected by Cathepsin D Activity Fluorometric Assay Kit. Results: Compared with the control group, the survival rates of SK-N-SH cells were decreased significantly in the 0.5-4.0 mmol/L MnCl(2) treatment groups (P<0.01) , the relative fluorescence intensities of 0.5 and 1.0 mmol/L MnCl(2) treatment groups were increased (P<0.01) . Compared with the control group, the 0.125-0.5 mmol/L MnCl(2) treatment groups had significant increase in the the expression of LAMP1 (P<0.01) . Compared with the control group, the expression of m-CTSD was significantly increased at the does of 0.125-0.25 mmol/L MnCl(2), while it was decreased at the does of 1.0 mmol/L (P<0.01) . Otherwise, it wasn't observed significant difference of the activity of CTSD between different MnCl(2) treatment groups. Conclusion: MnCl(2) could cause cytotoxicity in SK-N-SH cells. Lysosomes may play a normal function at low doses of manganese, but they may be damaged at high doses of manganese. As an organelle that can degradate substrates in autophagy, lysosomes participate in the neurotoxic mechanism of manganese.

Validation of the Malay version of the Amsterdam Preoperative Anxiety and Information Scale (APAIS).

BACKGROUND: Preoperative anxiety is a significant problem worldwide that may affect patients' surgical outcome. By using a simple and reliable tool such as the Amsterdam Preoperative Anxiety and Information Scale (APAIS), anaesthesiologists would be able to assess preoperative anxiety adequately and accurately. OBJECTIVE: The purpose of this study was to develop and validate the Malay version of APAIS (Malay-APAIS), and assess the factors associated with higher anxiety scores. METHODS: The authors performed forward and backward translation of APAIS into Malay and then tested on 200 patients in the anaesthetic clinic of University Malaya Medical Centre. Psychometric analysis was performed with factor analysis, internal consistency and correlation with Spielberger's State-Trait Anxiety Inventory (STAI-state). RESULTS: A good correlation was shown with STAI-state (r = 0.59). Anxiety and need for information both emerged with high internal consistency (Cronbach's alpha 0.93 and 0.90 respectively). Female gender, surgery with a higher risk and need for information were found to be associated with higher anxiety scores. On the other hand, previous experience with surgery had lower need for information. CONCLUSION: The Malay-APAIS is a valid and reliable tool for the assessment of patients' preoperative anxiety and their need for information. By understanding and measuring patient's concerns objectively, the perioperative management will improve to a much higher standard of care.

Expression of anti-neuroexcitation peptide III of scorpion Buthus martensii Karsch BmK ANEP III in plants.

Anti-neuroexcitation peptide III of Buthus martensii Karsch (BmK ANEP III) has better anti-epileptic and anticonvulsive effects in the test animal models. The present study is aimed at developing transgenic tomato and tobacco lines overproducing the ANEP III protein. Using the molecular cloning technique, the plant expression vector pBI-ANEP III was constructed successfully. The ANEP III expression cassette included a double CaMV 35S promoter with omega enhancers, the ANEP III gene with the Kozak sequence, the ER retention signal and the NOS terminator. Recombinant plasmids were transferred into Agrobacterium tumefaciens EHA105 by freeze-thaw transformation methods. By the Agrobacterium-mediated leaf disc transformation method, tobacco (Nicotiana tabacum) and tomato (Lycopersicum esculentum) lines were transformed. Transformants were screened and confirmed by PCR, RT-PCR and western blotting analysis. It was demonstrated that the ANEP III gene was successfully expressed in the genomic DNA of transgenic plants. The ANEP III protein was detected by immunofluorescence analysis, and the results confirmed the high amount of ANEP III protein, being 0.81 and 1.08% of total soluble proteins in transgenic tobacco and tomato. The study of plants with high expression levels of ANEP III has an important theoretical and practical significance and provides valuable information for establishing a new, economical and effective system for industrial protein production.

Development of the Video Analysis Scale of Engagement (VASE) for people with advanced dementia.

Background: The current study sought to develop a valid, reliable and unobtrusive tablet computer-based observational measure to assess engagement of people with advanced dementia. The Video Analysis Scale of Engagement (VASE) was designed to enable the rating of moment-by-moment changes in engagement during an activity, which would be useful for both future research and current residential care. Methods: An initial version of the VASE was tested. Face validity and content validity were assessed to validate an operational definition of engagement and develop an acceptable protocol for the scale. Thirty-seven non-professional and professional volunteers were recruited to view and rate level of engagement in music activities using the VASE. Results: An inter-class coefficient (ICC) test gave a high level of rating agreement across professionals and non-professionals.  However, the ICC results of within-professionals were mixed. Linear mixed modelling suggested that the types of interventions (active or passive music listening), the particular intervention session being rated, time period of video and the age of raters could affect the ratings. Conclusions: Results suggested that raters used the VASE in a dynamic fashion and that the measure was able to distinguish between interventions. Further investigation and adjustments are warranted for this to be considered a valid and reliable scale in the measurement of engagement of people with advanced dementia in a residential care setting.

Novel concepts in modification of radiation sensitivity.

PURPOSE: To determine whether biological effects of radiation, such as apoptosis, that differ from classical clonogenic cell killing, can be modified with agents that would not be expected to modify classical clonogenic cell killing. This would expand the range of potential modifiers of radiation therapy. METHODS AND MATERIALS: EL4 murine lymphoma cell apoptosis was determined by electrophoretic analysis of deoxyribonucleic acid (DNA) fragmentation. DNA was extracted 24 h after irradiation or addition of inducing agents. Modifiers of radiation-induced apoptosis were added immediately after irradiation. The effects of radiation on wounded endothelial monolayers were studied by scraping a line across the monolayer 30 min after irradiation. Cell detachment was used as an endpoint to determine the protective effect of prolonged exposure to retinol prior to irradiation. RESULTS: EL4 cell apoptosis can be induced by tert-butyl hydroperoxide or the glutathione oxidant SR-4077. Radiation-induced EL4 cell apoptosis can be inhibited with 3-aminobenzamide, an agent that sensitizes cells to classical clonogenic cell killing. Radiation-induced endothelial cell detachment from confluent monolayers can be modified by pretreatment with retinol. CONCLUSION: These results raise the possibility that radiation could induce apoptosis by an oxidative stress mechanism that is different from that involved in classical clonogenic cell killing. These and other recent findings encourage the notion that differential modification of classical clonogenic cell killing and other important endpoints of radiation action may be possible.

Pharmacodynamics of prolonged treatment with L,S-buthionine sulfoximine.

PURPOSE: To develop dosing criteria for the use of L-buthionine-S-sulfoximine (active diastereoisomer) as a glutathione depletor in the clinic, using a pharmacodynamic and pharmacokinetic in vitro-in vivo approach. METHODS AND MATERIALS: In vitro: L-buthionine-S-sulfoximine uptake was determined in human glioblastoma cells (T98G) and NIH-3T3 cells using 35S-labeled drug. Dose response relationships were derived for inhibition of glutathione synthesis in CHO cells, and for depletion of glutathione in exponentially growing T98G and CHO cells, as a function of extracellular L-buthionine-S-sulfoximine concentration. Steady-state glutathione levels for CHO and NIH-3T3 cells were measured using an enzymatic assay, while glutathione synthesis rates in CHO cells were determined using a flow cytometric assay. In vivo: L-buthionine-S-sulfoximine biodistribution was determined in male nude mice carrying human glioblastomas (T98G) intracranially, using 35S-labeled drug infused subcutaneously by osmotic pump. Tissue glutathione levels were measured using an enzymatic assay. RESULTS AND CONCLUSION: The observed cellular uptake t1/2 of approximately 55 min, coupled with a previously reported, rapid in vivo clearance of buthionine sulfoximine, suggest that continuous infusion would be preferable to bolus dosing. Effective concentrations of L-buthionine-S-sulfoximine (24 h exposure), required to lower cellular glutathione content to 50% of control (EC50), were under 1 mM for both cell lines. The amount of L-buthionine-S-sulfoximine in tissues (estimated from 35S drug disposition) reached steady state within 8 h and was proportional to the rate of infusion. Brain tumors were depleted to approximately 50% of control glutathione by a infusion rate of 0.25 mumoles/h (25 g mice). At lower infusion rates an increase in glutathione content was noted in certain nude mouse tissues including brain tumor xenografts.

Assessing data quality for decision support--emphasis on secondary analysis.

In secondary analysis, the use of available data makes it possible for the researcher to bypass the most time-consuming and costly steps in the research process. However, there are some noteworthy pitfalls and problems in working with existing data, especially the uncertainty of data quality. If the integrity and quality of data are not assured, statistical analysis of the data will not be reliable, no matter what statistical procedure is used. If the analysis is not reliable, the information used for decision support will not be accurate.

Are patients with hyperlipidemia being treated? Investigation of cholesterol treatment practices in an HMO primary care setting.

BACKGROUND: The main study objective was to investigate cholesterol treatment practices of primary care physicians in a managed care setting. METHODS: The study was a retrospective review of data with a quasiexperimental design. The National Cholesterol Education Program-Adult Treatment Panel II (NCEP-ATP II) guidelines were used as the reference for conducting a measurement model in the study. Data were randomly selected via a systematic probability sampling method from a health maintenance organization (HMO) capitated risk-based contracting medical clinic in southern Florida. RESULTS: Of the 348 patients selected for the study, 224 (65%) needed either dietary therapy (n = 106) or drug therapy (n = 118). However, only 16 patients (13.6%) had ever had cholesterol-lowering drug regimens prescribed during the 5-year study period. CONCLUSIONS: Our findings indicate that (1) primary care physicians have poorly adopted the cholesterol management practice recommended by NCEP guidelines and need to improve their recognition and treatment of hypercholesterolemia; and (2) the problem of underutilizing prescription medications may be associated with risk-sharing capitation arrangements between physicians and third-party insurers.

[Hereditary gingival fibromatosis: a case report].

Hereditary gingival fibromatosis is an uncommon congenital anomaly of undetermined etiology, the condition is manifested as a dense, diffuse, smooth or nodular overgrowth of the gingival tissue, usually begins with the eruption of the permanent incisors. However, it may happen as early as the eruption of the deciduous teeth. The case, a 30 years old woman with normal stature, visited our dental clinic because of "overgrown gum" which greatly affected mastication and appearance. A review of her past medical history revealed no relevant diseases. Oral examination showed gross fibrous enlargement of gingival without inflammation or swelling. Her father, two sisters and one daughter also suffered from this disease. Microscopically, there is a obvious increase in the amount of connective tissue that is relatively avascular and consists of densely arranged collagen bundles and numerous fibroblasts. The surface epithelium is thickened and acanthotic with elongated rete pegs. Gingivectomy with flap operation were performed to correct the gingival overgrowth. After periodontal treatment, two fixed partial dentures were constructed to replace the missing right upper and lower first molars. One year after operation, no obvious recurrence was noted clinically.

Study of prevalence, severity, and etiological factors associated with acute pancreatitis in patients infected with human immunodeficiency virus.

OBJECTIVES: Patients with the human immunodeficiency virus (HIV) disease can develop pancreatic gland inflammation from HIV infection and related causes, or from factors totally independent of it. The incidence and severity acute pancreatitis in patients with HIV diseases and the frequency of associated etiological factors have not been examined in any detail. The purpose of this study was to (a) determine the prevalence of acute pancreatitis, (b) evaluate severity of pancreatic gland inflammation, (c) identify commonly associated etiological factors with acute pancreatitis, and (d) examine the relationship between CD4 lymphocyte counts and serum pancreatic enzyme levels (amylase and lipase) in patients with HIV disease. METHODS: We examined the medical records of 321 patients with HIV disease seen at Sinai Hospital of Baltimore between July of 1993 to June of 1994. Data collected from these records included clinical, laboratory, and radiologic features of pancreatitis, staging of HIV disease, risk factors, CD4 lymphocyte counts, medications associated with the presence of opportunistic infections, Kaposi's sarcoma, and lymphoma. RESULTS: From 321 patients with HIV disease, 45 patients developed at least one episode of acute pancreatitis as defined by clinical and laboratory criteria during the 1-yr period. A statistically significant negative correlation was found between serum pancreatic enzyme level and the number of CD4 lymphocytes (r = -0.15, p < 0.05 for serum amylase; r = -0.2, p < 0.05 for serum lipase). Furthermore, patients with asymptomatic HIV infection or CD4 lymphocyte count >500 mm3 did not develop asymptomatic hyperamylasemia or acute pancreatitis. Furthermore, the presence of gallstones, active injection drug use, pentamidine therapy, Pneumocystis carinii, Mycobacterium avium intracellulare correlated significantly (p < 0.001) with the diagnosis of acute pancreatitis. CONCLUSIONS: A detailed review of medical records of patients with HIV disease seen in a community hospital in 1 yr (1993-1994) suggests a high incidence (14%) of mild to moderately severe acute pancreatitis. In this group of patients, pancreatic gland inflammation is commonly associated with gallstones, intravenous drug abuse, pentamidine intake, and Pneumocystis carinii and Mycobacterium avium intracellulare infections. In addition, marked reduction in CD4 lymphocyte count is associated with increase in serum pancreatic enzyme levels (amylase, lipase activity) suggesting pancreatic gland inflammation or altered pancreatic enzyme turnover.

Clinical and cost effectiveness of a new hepatocellular MRI contrast agent, mangafodipir trisodium, in the preoperative assessment of liver resectability.

BACKGROUND: Improved preoperative assessment of focal liver disease and tumors could have a potentially significant impact on their treatment. Mangafodipir trisodium (Teslascan; Nycomed Amersham Imaging, Little Chalfont, UK) is a new hepatocellular contrast agent for use with state-of-the-art MR imaging that, in early reports, is accurate in detection and characterization of liver lesions. METHODS: Records and diagnostic images of all patients undergoing enhanced Teslascan MRI (T-MRI) at our institution were reviewed. We assessed the relative sensitivities of contrast-enhanced CT scan (CECT) and T-MRI in detecting lesions, as well as the impact of T-MRI in the decision to operate or not on patients. In those patients taken to surgery, the correlation between T-MRI and intraoperative palpation and intraoperative ultrasound (IOUS) was determined. RESULTS: Fifty-four patients were noted on CECT to have focal liver lesions and subsequently underwent imaging with T-MRI. The T-MRI correlated with CT findings in 22 patients (41%), upstaged the liver disease in 26, and demonstrated fewer lesions in 6. Only 43 patients were considered operative candidates and T-MRI influenced the operative decision in 32 patients (74%), dissuading operative intervention in 14. In the 25 patients without clear preoperative evidence of unresectability who were taken to the operating room, T-MRI correlated with findings of intraoperative palpation in 19 (76%). In the 20 patients who underwent IOUS, T-MRI correlated with IOUS in 14 patients (70%). IOUS detected an additional nine lesions, all of which were <1 cm. Seventeen patients underwent resection and/or ablation of their liver lesions. Compared with pathology, sensitivities of CECT, T-MRI, and intraoperative evaluation were 61%, 83%, and 93%, respectively. T-MRI failed to predict hepatic-specific unresectability in only one of eight patients, the other seven having extrahepatic disease. CONCLUSIONS: These findings suggest that T-MRI is more sensitive than CECT in the preoperative predicting of the resectability of hepatic lesions. Despite T-MRI accurately correlating with intraoperative surgical findings, IOUS should be performed on all patients prior to a final decision to resect or ablate a focal liver lesion.

Studies on avian erythrocyte metabolism. XVI. Accumulation of 2,3-bisphosphoglycerate with shifts in oxygen affinity of chicken erythrocytes.

The ability of the chicken erythrocyte to accumulate 2,3-bisphosphoglycerate (2,3-P2-glycerate) and its effect upon the oxygen affinity (P50) of the cell suspensions have been determined. Erythrocytes from chick embryos, which contain 4-6 mM 2,3-P2-glycerate, and from chickens at various ages, which contain 3-4 mM inositol pentakisphosphate but no 2,3-P2-glycerate, were incubated with inosine, pyruvate, and inorganic phosphate. Red blood cells from 20-day chick embryos incubated in Krebs-Ringer, pH 7.45, containing 20 mM inosine and 20 mM pyruvate had an increase in 2,3-P2-glycerate from 4.3 to 11.9 mM after 4 h. Importantly, as 2,3-P2-glycerate concentration increased there was a corresponding increase in P50 of the cell suspension. Further, erythrocytes from 9- and 11-week, and 7-, 14-, 24-, and 28-month-old chickens when incubated similarly with inosine and pyruvate accumulated 2,3-P2-glycerate with corresponding increases in P50 of the cell suspensions. The ability of the red cell to accumulate this compound under the incubation conditions used apparently decreases with age of the bird (e.g., 11.9 mM in the 20-day embryo to 1.1 mM in the 28-month-old chicken after 4 h incubation). Despite the presence of significant amounts of inositol-P5, the accumulation of 2,3-P2-glycerate markedly decreases oxygen affinity of the cell suspensions. The delta P50/mumol increase in 2,3-P2-glycerate in the red cells of the 20-day chick embryo after 4 h incubation is 1.5 Torr; conversely, the delta P50/mumol decrease in 2,3-P2-glycerate in the red cells of the 17-day embryo after 6 h incubation in the presence of sodium bisulfite is 2.8 Torr. The demonstrated ability of the chicken erythrocyte to accumulate 2,3-P2-glycerate in response to certain substrates suggests that regulation of concentration of this compound could contribute significantly to regulation of blood oxygen affinity in birds.

Studies on avian erythrocyte metabolism. XVII. Kinetics and transport properties of myo-inositol in chicken reticulocytes.

The uptake of myo-inositol was determined in a reticulocyte-enriched fraction prepared from chicken blood and compared with uptake in mature erythrocytes. While reticulocytes accumulated inositol at levels more than threefold that of the plasma concentration, erythrocyte levels were only slightly higher than that of the plasma concentration. The rate of uptake in reticulocytes was approximately 66 mumol/ml rbc/h compared to 5 mumol/ml rbc/h in mature erythrocytes when measured at an inositol medium concentration of 250 microM. The kinetic analysis of inositol influx by reticulocytes reveals a two component system: saturable and nonsaturable. The saturable component, which has a Km for inositol of approximately 222 microM, is Na-dependent. This Na-dependent saturable component, which presumably reflects active transport of inositol, accounts for 30-35% of the transport process. The saturable component is completely inhibited by amiloride but to a lesser extent by ouabain and bumetanide. Moreover, in the course of reticulocyte maturation, the saturable component is lost concomitantly with the completion of the synthesis of myo-inositol pentakisphosphate and the drastic decrease in the membrane permeability to inositol. In addition, phloretin and cytochalasin B, which bind to hexose carriers and inhibit hexose sugar transport, also inhibited inositol transport. The uptake of inositol was not affected by excesses of 3-O-methylglucose (100 mM) or by physiological concentrations of D-glucose. Thus, the transport mechanism of myo-inositol appears distinct from that of D-glucose.