Objective:To investigate the influence of combined application of histone deacetylase inhibitor(HDACi)MS-275 and 5-FU on the apoptosis and cycle arrest on human hepatoma cell line HepG2 and unclose the related mechanism.Methods:Divided all the cells into 4 groups:control group,MS-275 group,5-FU group and drug combination group.Flow cytometry(FCM)was used to examine the effects of 5-FU with MS-275 on the apoptosis and cell cycle of HepG2 cells.Bcl-2,Bax,CyclinD1 and P21 protein were determined by Western blot assay.Results:5-FU and MS-275 combination could inhibit HepG2 cell growth through G0-G1 arrest,and induce apoptosis,both time and dose dependently.The combination of two agents increased P21 protein levels and decreased Bcl-2,CyclinD1 protein levels.The levels of Bax protein were not changed.Conclusion:The combination of 5-FU and MS-275 can induce apoptosis and cell cycle arrest,the effect might be associated with down-regulating expression of Cyclin D1 and Bcl-2 protein and upregulating the expression of P21 protein.

Objectives To investigate the expression of pax-6 in retina of infant monkeys with myopia induced by optical defocus, and to determine the role of pax-6 would play or not in onset and development of myopia and emmetropization. Methods Nine healthy infant rhesus monkeys, aged from 1 to 3 months, were selected and wore spectacle lenses or underwent photorefractive keratectomy (PRK).Transcription polymerase chain reaction method and quantitative analysis were used to determine the expression of pax-6 in the retina with myopia induced by optical defocus in different time, and the result was compared with that in retina without myopia. Results The myopia caused by hyperopic defocus was found. The expression of pax-6 in the retina with myopia induced by optical defocus was significantly higher than that in the retina without myopia(t=3.480,P=0.004). Conclusions The expression of pax-6 is enhanced by hyperopic defocus in the infant monkey retina, which suggests that pax-6 may be involved in vision-dependent eye growth and emmetropization.

Objective To summarize the published studies of diabetic cerebral infarction treated by integration of traditional and western medicine (ITWM) or pure western medicine and compare their clinical curative effects. Method The clinical study of ITWM for diabetic cerebral infarction were collected and seleted according to the standard. The effect of ITWM and only western medicine was compared, and Meta-analysis was made. Result 59 studies were included to analysis neurological deficit and the total efficiency. Meta-analysis revealed that the improvement of ITWM is better than pure western medicine treatment. Conclusion The existed limited evidences suggest that ITWM treatment of diabetic cerebral infarction can significantly improve neurological deficit and curative effect. More high-quality randomized, double-blind, placebo-controlled trials are needed to confirm the efficacy and safety of ITWM treatment of diabetic cerebral infarction.

Objective To establish a system in which mouse embryonic stem cells (mESCs) differentiate into insulin-secreting cells in vitro. Methods mESCs were cultured to form embryoid bodies(EBs). EBs were cultured in serum-free medium to obtain nestin-positive cells. The selected nestin-positive cells were expanded,then added nicotinamide into the medium to induce the differentiation of nestin-positive cells. Immunochemistry staining and flow cytometry analysis were made on 15th day after induction.Results Flow cytometry analysis revealed that the percentage of nestin-positive cells were different from EBs. The percentage of nestin-positive cells from EBs were higher than those of other diameters. Nestin-positive cells induced by nicotinamide formed islet-like cell clusters. Percentage of insulin-positive cell induced by 10 mmol/L nicotinamide was higher than those induced by 0 mmol/L or 5 mmol/L nicotinamide(P

Heart Neoplasms ; pathology ; Humans ; Solitary Fibrous Tumors ; pathology ; Tumor Burden

0.05), but did inhibit activity of iNOS (P

BACKGROUND: Umbilical cord blood (UCB) with limited karyocytes is mainly used in child patients. Recently, physicians have tried to mix two units of cord blood in the treatment of adults with hematological system diseases.OBJECTIVE: To monitor quantitatively the dynamic changes and the development rules of engraftment, chimera types and relative amount after allogeneic transplantation of mixed UCB from two units in adults with leukemia.DESIGN: Donors and the recipient were regarded as observational subjects in umbilical cord blood transplantation (UCBT). DNA extracted from blood samples of donors and the recipient before and after transplantation was considered as detecting samples. Short tandem repeat (STR) loci were as observational measures.SETTING: Key Laboratory of Immunology and Genetics of Institute of Transfusion Medicine of Shenzhen Blood Center.PARTICIPANT: A 43-year male patient with acute myeloid leukemia (AML), 75 kg, who was hospitalized at Shenzhen Hospital of Peking University, was enrolled in June 2005. The patient received two units of human leucocyte antigen (HLA), one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2) at month 6 after complete remission from first chemotherapy. UCB was collected from Guangzhou umbilical cord blood bank. The patient signed the informed consent.METHODS: The adult with AML received two units of HLA, one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2). Nine STR loci of the blood sample were determined before and after transplantation by quantitative technique of fluorescence labeling with multiplex polymerase chain reaction (MPCR), while the engraftment and chimera types were qualitatively evaluated by comparing differential loci between the recipient and the donors. The relative amount of two units of UCB was calculated in the patient after transplantation according to the differential gene peak areas of two donors with 377XL DNA sequencer after fluorescence scanning. The engraftment level and the development rules of donors' cells were analyzed quantitatively. In addition, the results were also compared with that of HLA loci distinct analysis for engraftment.MAIN OUTCOME MEASURES: After UCBT, transition process of nine STR loci of the recipient and two donors was observed, and engraftment was quantitatively and qualitatively described.RESULTS: Two units of UCB at day 15 after transplantation were engrafted simultaneously and revealed a complete chimera of the two. The relative amounts of UCB 1 and UCB 2 were 51.3% and 48.7%, respectively. Subsequently, UCB 1 went up to 70.0% and UCB 2 declined to 30.0% at day 30. However, only the genotype of UCB 1 was detected at day 52, and engraftment turned to a complete chimera of a single donor. The one with fewer karyocytes was rejected and the one with more karyocytes was engrafted for a long term.CONCLUSION: To detect quantitatively STR chimera with fluorescence labeling and MPCR can show precisely the engraftment level and the change of two units of UCB. It provides an accurate and reliable experimental basis for clinical UCB application and donor selection. It is proved that adult transplantation at the same time with mixed UCB from two units HLA one locus mismatched unrelated donors is feasible.

Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Tianjihuang gran -ules.Methods:A ZORBAX Eclipse XDB-C18 column was used.The mobile phase was methanol-0.2%phosphoric acid solution (42∶58).The detection wavelength was 350 nm and the flow rate was 1.0 ml· min-1.Results:The linear range of isoquercitrin, quer-citrin and quercetin was 4.572-114.300 μg· ml-1(r=0.999 8), 7.028-175.700 μg· ml-1(r=0.999 8) and 2.195-54.880 μg· ml-1(r=0.999 9), respectively.The average recovery was 98.01%(RSD=1.24%, n=6), 98.14%(RSD=0.98%, n=6) and 97.00%(RSD=1.09%, n=6) , respectively.Conclusion:The method is specific and simple,which can be used for the quality con-trol of Tianjihuang granules.

Objective To explore the lung developmental disorder of rats with gestational diabetes mellitus (GDM) via investigating the GDM rat fetal lung structures and expression of pulmonary surfactant proteins (SP)-B,SP-C,thyroid transcription factor (TTF)-1 and pleiomorphic adenoma gene like (PLAGL)-2.Methods Sprague-Dawley rats were used to construct the GDM model.Twenty GDM rats were used as GDM group and 20 normal pregnant rats as control group.Cesarean section was performed on day 21 of gestation and random blood sugar was detected,and fetal rats were counted and weighed.Ultrastructure of the fetal lungs was studied by transmission electron microscopy.Sixty fetal rats were selected randomly in each group,and 360 paraffin sections were made from fetal lungs.One hundred discontinuous paraffin sections were picked up in each group to observe morphological and structural changes under optical microscope.The other one hundred discontinuous paraffin sections were picked up in each group to detect the location and expression of SP-B,SP-C,TTF-1 and PLAGL-2 protein by immunohistochemistry.Nine fetal rats were selected randomly to detect the expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in fetal lung tissues by Western blotting.Twenty seven fetal rats were selected randomly to detect the mRNAs level of SP-B,SP-C,TTF-1 and PLAGL-2 by real-time quantitative polymerase chain reaction.Independent sample t-test was used for statistical analysis.Results The average random blood glucose level in GDM group was significantly higher than that in control group [(26.8± 2.8) vs (4.9± 0.5) mmol/L,t=-34.05,P=0.00].The average weight of fetal rats in GDM group was higher than that in control group [(5.6±0.6) vs (5.2±0.5) g,t=-1.97,P=0.03].Alveolar number (10.1 ± 1.6 vs 12.1 ± 1.3) and alveolar area [(986.9 ± 5.5) vs (1 257.3± 5.0) μ m2] in GDM group was less than that in control group (t=9.84 and 27.53,both P ＜ 0.05).Alveolar septum [(11.5±6.2) vs (9.9±4.3) μm] in GDM group was higher than that in control group (t=-2.17,P ＜ 0.05).Microvillus in type] cells were short and the number of lamellar bodies was significantly decreased in GDM group.SP-B,SP-C,TTF-1 and PLAGL-2 proteins were distributed in the cytoplasm in granular form.The average value of absorbance of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 1.15±0.12,1.23±0.06,0.87±0.21 and 1.21 ±0.18 respectively;and that in control group was 1.22±0.05,1.31 ±0.14,1.12±0.09 and 1.33 ±0.07 respectively.The value in GDM group was lower than that in control group (t=2.40,2.35,4.89,and 2.77 respectively,all P ＜ 0.01).The expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 0.57± 0.09,0.45±0.03,1.50±0.04 and 1.11 ±0.04 respectively;and that in control group was 0.81 ±0.03,0.66±0.04,1.69±0.05 and 1.46±0.07 respectively.The value in GDM group was lower than that in control group (t=1 1.77,11.09,8.80 and 13.37,respectively,all P ＜ 0.01).The mRNA level of SP-B,SP-C,TTF-1 and PLAGL-2 in GDM group was 0.60±0.04,0.79±0.04,0.81 ±0.03 and 0.79±0.05 respectively;and that in control group was 1.06±0.19,1.03±0.24,1.03±0.18 and 1.02±0.19 respectively.The value in GDM group was lower than that in control group (t=6.80,2.98,3.54 and 3.54 respectively,all P ＜ 0.01).Conclusions The protein expression level of SP-B and SP-C in fetal lungs of GDM rats decreases obviously,possibly because of the down-regulation of the gene expression of TTF-1 and/or PLAGL-2.The pathological changes in fetal lungs of GDM rats might be associated with the descending level of SP-B and SP-C protein.

Aim To investigate the protective effect of ginsenoside Rb1 against apoptosis induced by H_2O_2. Methods H_2O_2 was used to build an oxidative stress-induced injury model in neonatal rat cardiomyocytes. After treated with gensenoside Rb1(20, 40, 80 mg?L -1),the apoptosis rate, the content of malondialdehyde (MDA), and the activity of superoxide dimutase (SOD) of the cardiomyocytes were examined. The intracellular calcium indicated by the fluorescence in cells were measured by the laser confocal microscope. Results Compared with the model group, the apoptosis rate and the content of MDA of the cardiomyocytes decreased greatly (P