Objective:To discuss the effect and mechanism of autophagy inhibitor 3-MA on arsenic trioxide inducing apoptosis of acute T-cell leukemia cell line Jurkat cells.Methods:Proliferation inhibition of Jurkat cells treated with arsenic trioxide was detected by XTT.Morphological characteristics of Jurkat cells treated with different concentrations arsenic trioxide were observed by electron mi-croscope.Microtubule-associated protein 1 light chain 3B (LC-3B) protein expression was detected by Western blot and flow cytome-try.Apoptosis rates of Jurkat cells treated with 3-MA combining arsenic trioxide were detected by flow cytometry using AnnexinV-FITC/PI double staining.Results:Arsenic trioxide inhibited the growth of Jurkat cells in a dose and time dependence.We observed different morphological characteristics of autophagy , apoptosis and necrosis accompanying more autophagosomes in Jurkat cells which were treated with arsenic trioxide 2.5,5,10 μmol/L after 24 h.LC3B mean fluorescence intensity (MFI)relative multiples were(3.1±0.2) fold,(4.6±0.31)fold,(34.2±4.5)fold with 5 μmol/L arsenic trioxide treated Jurkat cells 0,24,48 h,and the P values between each of the two groups were less than 0.05,which increased depending time consistently with the growth inhibition rates.LC-3B protein expression gradually increased treated Jurkat cells with arsenic trioxide after 24 h,48 h.The growth inhibition rate (60.6±8.3)%was significantly different treated with arsenic trioxide combining 3-methyl adenine ( 3-MA ) while it was ( 33.4 ±9.1 )% treated with arsenic trioxide alone, however, LC-3B protein expression gradually decreased.Jurkat cell apoptosis rate ( 44.96 ±3.60 )% was significantly increased treated with arsenic trioxide combining autophagy inhibitor(3-MA) while it was (2.94±0.26)% treated with arsenic trioxide alone, and this difference was statistically significant.Conclusion: 3-MA increased apoptosis rates of Jurkat cells inducing by Arsenic trioxide and it may be related with inhibition of autophagy and induction of apoptosis.

Background Researches have demonstrated that ocular hypertension induces the ischemia-reperfusion of retina and further leads to the degeneration of retinal ganglion cells,but its mechanism is beyond understanding.Objective The present study aims to observe the effects of static pressure on the morphology,proliferation activity and viability of cultured retinal microvascular endothelial cells (RMECs) and evaluate the expression of ET-1 and NO in these cells under variant static pressure.Methods RMECs were isolated from 30 healthy Wistar rats and cultured using explant culture method by Ⅷ factor antibody and PECAM-1 antibody.The static pressure of 1.33kPa,2.67kPa,5.33kPa and 10.67kPa was used in culture bottle respectively.The RMECs without static pressure were used as normal control group.The morphology of RMECs under the different static pressure was observed by inverted phase contrast microscopy,and the number of RMECs was counted using the counting plate.Cellular viability was studied by trypan blue staining.The changes of ET-1 and NO_2~-/NO_3~-,two metabolic products of NO,in the medium were detected by radioimmunoassay and Griess's nitrate reductase method.The expression of ET-1,eNOS and iNOS mRNA in RMECs was analyzed by semi-quantitative RT-PCR 24 hours after treatment of variant static pressure.Results Cultured RMECs sticked well at 24 hours and reached to confluence at 48 hours and showed the red fluorescence for Ⅷ factor antibody and PECAM-1 antibody.Enlargement of nuclei,extenders of cell bodies and suspension of RMECs in medium were observed.The number of RMECs was gradually increased.The cell viability was reduced with the raise of static pressure among these four groups(F=12.205,P＜0.01;F=11.180,P＜0.01).The static pressure increased the content of ET-1 released by RMECs in 2.67kPa,5.33kPa and 10.67kPa of static pressure groups,and concentrations of NO_2~-/NO_3~- in the medium showed a significant increase in 5.33kPa and 10.67kPa of static pressure groups compared with normal and 1.33kPa of static pressure groups(P＜0.01).The expressions of ET-1 mRNA,eNOS mRNA and iNOS mRNA were considerably enhanced in 5.33kPa and 10.67kPa of static pressure groups compared with normal control group(P＜0.01).Conclusion Raised static pressure causes the alteration of RMGCs structure and morphology.Static pressure could upregulate the expressions of ET-1,eNOS and iNOS mRNA in RMECs and increase the release of ET-1 and NO.This pathway might be one of pathologic mechanisms of retinal injury induced by high intraocular pressure.

OBJECTIVE:To revise the accreditation standards of the national clinical pharmacy key specialty,promote the nor-malization and standardization of clinical pharmacy services and improve the service level of clinical pharmacy in our country. METHODS:Based on JCI international hospital accreditation ideas,suggestions for Clinical Pharmacy State Clinical Key Program Construction Score Standard for trial implementation (900 points) published by former Ministry of Health in 2012,were put for-ward. RESULTS & CONCLUSIONS:For the problems that there were lack of pharmacy service institutional management,process control and quality management standards,some classification programs were not clear,and did not fully reflect the clinical phar-macy service quality and work effect evaluation requirements,corresponding revisions were proposed in terms of basic conditions, information management,clinical pharmacy work management system,etc. The revised standard pays more attention and refines the quality system construction in system,standard,process,quality control,effect evaluation,which can promote the enhance-ment of our clinical pharmacy service levels,normalization and standardization of clinical pharmacy services.

Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.
Animals ; CHO Cells ; Calcium Signaling ; Cricetinae ; Cricetulus ; Humans ; Pyrazoles ; pharmacology ; Quinolines ; pharmacology ; Receptors, Neurotensin ; antagonists & inhibitors ; genetics ; metabolism ; Transfection

Objective To investigate the effect of intravenous infusion of hyper-oxygenated solution (HOS) on small intestinal ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty-four rabbits of both sexes,weighing 2.5-3.2 kg,were randomly divided into 3 groups ( n =8 each):sham operation group (group S),I/R group,and HOS group.Small intestinal I/R was produced by clamping the superior mesenteric artery (SMA) for 1 h followed by 2 h of reperfusion in I/R and HOS groups,while the SMA was only clamped in group S.HOS was infused intravenously at a rate of 20 ml· kg-1 ·h -1 via the auricular vein starting from the time immediately after clamping the SMA in group HOS and the equal volume of normal saline was infused instead of HOS in group I/R.Blood samples were obtained from the inferior vena cava at 2 h of reperfusion to detect the concentration of serum lactic acid.The animals were then sacrificed and the small intestine was removed for determination of the malondialdehyde (MDA) content,and activities of superoxide dismutase (SOD),catalase (CAT) and glutathione peroxidase (GSH-Px) in intestinal tissues and for microscopic examination.The pathological changes of the intestinal epithelia were observed and the damage.to the mucous membrane was scored.The internal organs were removed and bacterial translocation from gut to the internal organs was observed.Results Compared with group S,the level of MDA and lactic acid,and rate of bacterial translocation were significantly increased,and the activities of SOD,CAT and GSH-Px were significantly decreased in groups I/R and HOS ( P ＜ 0.05).Compared with group I/R,the level of MDA and lactic acid,rate of bacterial translocation,and activities of SOD,CAT and GSH-Px were significantly decreased in group HOS ( P ＜ 0.05).Conclusion Intravenous infusion of HOS can reduce small intestinal I/R injury in rabbits.

AIM: To report one case of huge fungal granuloma in the deep maxillofacial region with its clinical findings,diagnosis and treatment process to improve our understanding of fungal granuloma and lessen misdiagnosis.METHODS: The literature was reviewed with combination of case report.RESULTS: The clinical and imaging findings of fungal granuloma had no specificity and were very subject to misdiagnosis. Confirmed diagnosis needed histopathological examination and bacterial culture.CONCLUSION: Deepened understanding of this disease can decrease misdiagnosis and benefit prompt and proper treatment.

A 2,4 -dichlorophenol degrading Pseudomonas strain GI241-1 was isolated from a soil sample. The dienelactone hydrolase gene, designated as dcpD which encodes dienelactone hydrolase involved in transforming cis-2-chloro-dienelactone into 2-chloromaleylacetic acid, was cloned from this bacterium strain. The gene cloning strategy was to construct genomic library after location of its neighbouring gene by Southem blot and to screen the aim transformant by dot blotting. Sequencing results showed that length of dcpD is 702bp. The sequence of dcpD and the deduced amino acid are different from the relative sequences registered in the GenBank.

Objective Standard neuropsychological assessment plus structural and functional imaging were used in accurate diagnosis of Benson's syndrome (posterior cortical atrophy).Method Serial neuropsychological screening and integrative assessments of visual spatial function, 3D structural MRIimaging and functional FDG-PET imaging were used in two cases of Benson' s syndrome.Results The clinical signs were agnosia, optic ataxia, apraxia, alexia, agraphia and prosopagnosia.MRI imaging revealed bilateral parietal and occipital lobe atrophy.FDG-PET imaging revealed low metabolism in the posterior cortex.The agraphia was constructive: the words were correct but written in the wrong location.Conclusion Standard neuropsychological assessments can recognize the disease nature.When combined with the structural and functional imaging, a correct diagnosis of Benson's syndrome can be made.

OBJECTIVETo investigate the relationship between lumbosacral transitional vertebra and the lumbar disc herniation (LDH).

METHODSThe X-ray photographs of lumbar vertebra were retrospectively studied of patients with and without LDH confirmed by surgery, furthermore, the differential incidence of LDH between the two groups and the relationship between transitional vertebra and the position of disc herniation were statistically analyzed.

RESULTSThe incidence of lumbosacral transitional vertebra was 18.3% in the control group, 52.7% in the LDH group, the difference was statistically significant. In the group with single lumbosacral transitional vertebra, there was 75.9% of LDH occurred on the same side of the transitional vertebra, 81.8% of which occurred at the upper one disc of the transitional vertebra; whereas 17.2% of LDH on the opposite side of the transitional vertebra, 80.0% of which occurred at the lower one disc of the transitional vertebra.

CONCLUSIONSThere is a closer relationship between lumbosacral transitional vertebra and the LDH, and the lumbosacral transitional vertebra is one of the important factors in the emergence of LDH.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Intervertebral Disc Displacement ; etiology ; Lumbar Vertebrae ; diagnostic imaging ; Male ; Middle Aged ; Radiography ; Sacrum ; diagnostic imaging ; Spondylolisthesis ; complications

[Purpose] To investigate the relation between isotonic leg extension strength (ILES) and the prevalence of metabolic syndrome (MS) in Japanese male adults.[Methods] This cross-sectional study included 395 Japanese men. Metabolic syndrome was determined according to the criteria of International Diabetes Federation (IDF), Japan Society for the Study of Obesity (JASSO), or National Cholesterol Education Program's Adults Treatment Panel III (NCEP-ATPIII). Muscular strength was measured on a horizontal dynamometer. Subjects were divided into tertile levels according to ILES (watts/kg). Multiple logistic regression analysis was used to examine the relation between ILES levels and the prevalence of MS.[Results] The prevalence of MS were 14.4% (IDF), 17.0%(JASSO), and 20.0% (NCEP-ATPIII). After adjustment for confounding factors, the odd's ratios (95% confidence interval) of MS (IDF) compared with the lowest ILES level were 0.90 (0.48-1.68) and 0.31 (0.13-0.64) in the middle and high levels of ILES (p for trend=0.03). The association between the level of ILES and the prevalence of MS by JASSO and NCEP-ATPIIIcriteria were similar.[Conclusion] The prevalence of MS was associated with the level of ILES.