Objective To investigate the therapeutic effects of arsenic trioxide(ATO) plus vitamin K2(VK2) on proliferation of HL-60 cells from acute promyelocytic leukemia cell line and explore the possible mechanism.Methods ①HL-60 cells were exposed to ATO(0.0,0.5,1.0,2.0,4.0 μmol/L),VK2(0.0,2.5,5.0,10.0,20.0μmol/L),or both of different concentrations (0.5 μmol/L ATO + 2.5 μmol/L VK2,1.0 μmol/L ATO + 5.0μmol/L VK2,2.0 μmol/L ATO + 10.0 μmol/L VK2,4.0 μmol/L ATO + 20.0 μmol/L VK2) for 24,48 or 72 h,respectively.The method of CCK-8 was used to assess the proliferation of HL-60 cells and the half inhibitory concentration(IC50) of ATO or VK2 was calculated,respectively.②Combination index (CI) was used to evaluate the combinative effect of the two treatments:CI ＜ 1,=1 or ＞ 1 indicated synergistic,additive,or antagonistic effect,respectively.③After HL-60 cells were treated with 1.0 μmol/L ATO or 5.0 μmol/L VK2 individually or simultaneously for 48 h,Annnexin V/PI staining was performed to identify the apoptosis rate of each group.Untreated cells were used as control group.Results ①ATO or VK2 alone inhibited the proliferation of HL-60 cells in a concentration and time dependent manner.The IC50 of ATO or VK2 at time of 24,48,72 h were (22.86 ± 2.44),(6.66 ± 0.34),(4.14 ± 0.41) and (18.40 ± 1.12),(13.48 ± 0.73),(8.95 ± 0.40) μmol/L,respectively; ②The combination of ATO and VK2 illustrated a synergistic effect with CI ＜ 1.③No statistical difference was found among control group [(4.38 ± 0.56)％],1.0 μmol/L ATO group [(5.76 ± 1.63)％] and 5.0 μmol/L VK2 group [(6.38 ± 1.42)％] in the apoptosis rate(all P ＞ 0.05).However,the apoptosis rate of combined group did rise to (44.18 ± 8.42)％,with a significant improvement to that of VK2 or ATO group alone (all P ＜ 0.01).Conclusions The combination of VK2 and ATO exhibits an enhanced synergistical inhibitive effect on proliferation of HL-60 cells,and apoptosis may be involved in this synergy in part.

Animals ; Cyanides ; toxicity ; Mice ; Random Allocation ; Toxicity Tests, Chronic

Objective To observe curative effect of olopatadine in treatment of allergic conjunctivitis combined with frequently winking in children. Design Prospective case series. Participants Ninety seven children with frequently winking aged 3-12 years in Shanghai Eye and ENT Hospital. Methods According to the factor in causation of frequently winking (FW), 97 children were divided into allergic conjunctivitis (AC) group, trichiasis combined AC group, AC with tics group, trichiasis group, tics group and unknown cau-sation group after clinical examination. The first 3 groups with AC were treated with olopatadine 0.1% ophthalmic solution b.I.d.(n=35) and emedastine 0.05% ophthalmic solution was used q.I.d, in AC group as control(n=32). The patients were followed after 3 weeks and the extent of symptoms of FW was recorded. Main Outcome Measures Extent of the symptoms of FW. Result The effective rate of olopatadine in treating FW caused by AC or AC combined with other diseases was 80.43%. The effective rate in FW children caused mainly by AC was 82.86% which was higher than that of control drug emedastine (75.0%) (P=0.02). Seven cure cases were all in olopatadine treating group. Conclusion AC is the most common and main causative factor in FW children. Early use of olopatadine ophthalmic drop can effectively alleviate FW symptoms. (Ophthalmol CHN, 2009, 18: 272-274)

Aged ; Humans ; Lung ; pathology ; Male ; Middle Aged ; Pneumoconiosis ; complications ; diagnosis ; pathology ; Reference Standards

OBJECTIVETo explore the relationship between gene expression of human telomerase reverse transcriptase (hTERT) and its clinical characteristics in leukemia.

METHODSThe protocol of RT-PCR was used to detect the hTERTmRNA expressing levels in peripheral blood samples from leukemic patients under primary treatment(n=42), in complete remission(n=21), with recurrent leukemia (n=4); and from normal subjects (n=5), respectively.

RESULTSThe positive percentage of hTERTmRNA expression was 73.81% for the primary treatment cases, and 19.05% for the complete remission cases. All of the recurrent cases gave positive results. One of the normal controls presented low level of hTERTmRNA expression. The expressing level of hTERTmRNA in primary treatment cases was 0.64+/-0.21, in complete remission leukemia 0.31+/-0.16, in recurrent cases 0.84+/-0.09, and in normal controls 0.10.

CONCLUSIONThe activation of telomerase may be an essential factor in the development of leukemia and usually be the late event in its progression. As an indicator of leukemia cell, the detection of hTERT mRNA may be used in clinical analysis, disease monitoring and prognosis judgement.

Acute Disease ; Adolescent ; Adult ; Child ; Child, Preschool ; DNA-Binding Proteins ; Female ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Infant ; Leukemia ; genetics ; pathology ; Male ; Neoplasm Recurrence, Local ; RNA, Messenger ; genetics ; metabolism ; Remission Induction ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics

The modulation of ACh on delayed rectifier-like potassium currents (I(K)) was studied in freshly dissociated cerebral cortical neurons using the whole-cell patch-clamp technique. Wistar rats between 10- and 14-day old of both sexes were used. After rats were decapitated, their brains were quickly removed, iced, and then manually cut into 400 mum slices. Slices were then incubated for 0.5 h at 32 degrees C in a buffered artificial cerebrospinal fluid (ACSF) bubbled with 95% O2, 5% CO2. Slices were then removed into buffered ACSF containing protease (0.5 mg/ml) at 32 degrees C. After 30 min of enzyme digestion, tissue was rinsed three times in the buffered saline. Then the enzyme-treated slices were mechanically dissociated with a graded series of fire-polished Pasteur pipettes. The cell suspension was then plated into a 35 mm dish and placed on the stage of a Olympus inverted microscope. For whole-cell recordings of currents, standard voltage-clamp techniques were used. Neurons were held at -80 mV, and the I(K) was evoked by 2 000 ms depolarizing voltage commands to potential between -40 mV and +60 mV in 10 mV steps applied at a frequency of 0.5 Hz. It was found that the inhibitory effect of ACh (0.1, 1, 10, 100 mumol/L) on I(K) was dose-dependent. It was also found that ACh affected the activation process of I(K) significantly, i.e., the activation curve of I(K) was characterized by half-activation potential of (-41.8+/-9.7) mV and a slope factor of (30.7+/-7.2) mV in the cortical neurons and they were changed to (-122.4+/-38.6) mV and (42.4+/-7.0) mV, respectively, after giving ACh (10 mumol/L). Tubocurarine (100 mumol/L) antagonized the inhibitory effect of ACh on I(K), and the drop of currents varied from the control value of (36.5+/-7..8)% to (16.9+/-13.8)% (n=8, P<0.01). 4-DAMP (10 mumol/L) blocked the inhibitory effect of ACh on I(K), and the currents reduced from the control value of (36.5+/-7.8)% to (26.8+/-4.7) % (n=6, P<0.05). Pirenzepin did not antagonize the inhibition of ACh on I(K) (n=7, P>0.05). Chelerythrine (20 mumol/L) blocked the inhibitory effect of ACh on I(K) and the currents reduced from the control value of (36.5+/-7.8)% to (11.7+/-17.3)% (n=6, P<0.05). On the contrary, PDBu (10 mumol/L) strengthened the inhibition of ACh on I(K) and the drop of currents changed from the control value of (36.5+/-7.8)% to (59.2+/-14.0)% (n=5, P<0.05). PDBu abolished the antagonism of chelerythrine on ACh in cortical neurons. It is suggested that the ACh-induced depolarization of neurons in the cortex is attributed to the inhibition of I(K) that is most likely evoked by the activation of nicotinic ACh receptors and muscarinic M3 receptor via protein kinase C (PKC) signal transduction pathway.
Acetylcholine ; physiology ; Animals ; Cell Separation ; Delayed Rectifier Potassium Channels ; antagonists & inhibitors ; Female ; Male ; Neurons ; metabolism ; physiology ; Patch-Clamp Techniques ; Protein Kinase C ; metabolism ; physiology ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; metabolism ; Receptors, Nicotinic ; metabolism ; Signal Transduction ; physiology ; Somatosensory Cortex ; cytology ; physiology

OBJECTIVETo study the genetic etiology of an autosomal dominant dentinogenesis imperfecta in a Chinese family.

METHODSThe molecular change of the disease in the family was analyzed through the clinical examination, linkage analysis, mutational screening of the DSPP gene and restriction fragment length polymorphism analysis.

RESULTSThe disease related gene was completely linked with microsatellite marker D4S1534. We found a novel mutation in the first exon of the DSPP gene (c.49C>T, p.Pro17Ser). All patients in the family had the mutation, while this mutation was not observed in the normal individuals of this family and 100 unrelated controls.

CONCLUSIONThe p.Pro17Ser identified in the family was a new pathogenic mutation. Our finding provided further understanding of the molecular mechanism of dentinogenesis imperfecta.

Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Dentinogenesis Imperfecta ; genetics ; Exons ; Extracellular Matrix Proteins ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Molecular Sequence Data ; Mutation ; Pedigree ; Phosphoproteins ; Sialoglycoproteins ; Young Adult

A new aporphine alkaloid (1), together with five known analogues (2-6), has been isolated from the branch of Litsea greenmaniana by using various chromatographic techniques. Their structures were identified by spectroscopic data analysis ( MS, IR, 1D and 2D NMR) as 2,9-dihydroxy-1,10-dimethoxy-4,5-dihydro-7-oxoaporphine (1), laurotetanine (2), N-methyllaurotetanine (3), isodomesticine (4), isocorydine (5), and norisocorydine (6). Compound 1 was a new compound, and compounds 2-6 were obtained from this plant for the first time.
Alkaloids ; chemistry ; Aporphines ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Litsea ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To study the role of inflammation in white matter damage of spontaneously hypertensive rats (SHRs) through observing the pathology changes of tissues after white matter damage and detecting the levels of inflammation-related indicators.Methods Eighteen 40-week-old male SHRs were chosen as experimental group,and seven male Wistar-Kyoto (WKY) rats were used as control group.The animal brain tissues were taken for hematoxylin-eosin staining (HE staining) and immunohistochemical staining to observe the pathological changes,and the levels of myelin basic protein (MBP),neurofilament (NF) and glial fibrillary acidic protein (GFAP).Real-time PCR was employed to detect toll-like receptor 4 (TLR-4),monocyte chemotactic protein 1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) expression levels in the white matter tissues.Results The white matter of 40-week-old SHRs was apparently injured.HE staining displayed sponge-like changes in the white matter and immunohistochemical staining showed astrocyte activation,reduced number of axonal and demyelination in the white matter.As compared with those in the WKY rats,TLR-4,MCP-1 and VCAM-1 mRNA expressions in SHR white matter were significantly increased (P＜0.05); and TLR-4,MCP-1 and VCAM-1 expression levels in SHRs were positively related to the degree of white matter damage.Conclusion The white matter damage in 40-week-old SHRs is similar to that of LA;inflammation is involved in the pathophysiological process of white matter damage,being one of induced factors of white matter injury.

This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Base Sequence ; Hemoglobin A2 ; immunology ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; alpha-Thalassemia ; immunology