Objective To study the effect of glucocorticoid on serum interleukin(IL)-1,IL-6,transforming growth factor-?_1(TGF-?_1) and tumor necrosis factor alpha(TNF-?) in children with primary nephrotic syndrome(PNS).Methods The levels of cytokines on serum were compared in 3 different patho-type,and IL-1,IL-6,TGF-?_1 and TNF-? in serum were detected using enzyme-linked immunosorbent assay(ELISA) in 25 children with PNS before and after treatment with glucocorticoids.Results Significant difference of IL-1,IL-6,TGF-?_1 and TNF-? were found between the two group before and after the treatment(P0.05).The level of TGF-?_1 in MCD was lo-wer than that in MsPNG,FSGS before and after the treatment(P0.05),and the level of it in FSGS was higher than that in MsPNG after the treatment(P

Objective To explore the effects of matrine and oxymatrine on apoptosis in human hepatocarcinoma SMMC-7721 cells.Methods The MTT assay and double staining of annexin V-fluorescein isothiocyanate(annexin V-FITC)/propidium iodide(propidium iodide, PI)were used to detect proliferation and apoptosis of SMMC-7721 cells, respectively.Results When the concentrations of matrine and oxymatrine were 0.50 mg/ml, 1.00 mg/ml and 2.00 mg/ml, the proliferation inhibition rates in SMMC-7721 cells was gradually increased in a dose- and time-dependent manner,and the inhibition of matrine on proliferation were greater than that of oxymatrine in the same concentration(1.00 mg/ml)(24 h:42.39%±0.04%vs. 21.36%±0.02%;48 h: 51.69%±0.03%vs. 36.16%±0.02%;72 h: 78.98%±0.05%vs. 61.24%±0.13%;allP<0.05). When the concentrations of matrine and oxymatrine were 0.25 mg/ml, 0.50 mg/ml and 1.00 mg/ml, the apoptosis rates of SMMC-7721 cells were significantly increased;and induction of matrine in apoptosis in SMMC-7721 cells was greater than that of oxymatrine at the same time point(48 h)(apoptosis rates in 0.25 mg/ml, 4.08%±0.20%vs. 2.20%±0.18%;0.50 mg/ml: 4.32%±0.19%vs. 3.08%±0.26%;1.00 mg/ml: 9.93%±0.18%vs. 9.01%±0.20%;allP<0.05).Conclusion Matrine and oxymatrine can inhibit proliferation and promote apoptosis human hepatocarcinoma SMMC-7721 cells.

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

[Objective] To establish an effective and stable method to induce hematopoietic cells from embryonic stem(ES) cells,the phenotype and function of ES-derived hematopoietic cells induced by stromal cell conditioned medium (SCCM) of yolk sac (YS),fetal liver (FL) or bone marrow (BM) were analyzed and compared.[Methods] 10% of YS-SCCM,FL-SCCM or BM-SCCM was added to culture system for differentiation of ES cells.Flow cytometric analysis was used to identify expression of Flk1,Integrin α4,Sca-1,and CD34.Colony analysis was used to identify the quantity of high proliferative potential colony-forming cells (HPP-CFC) in differentiated ES cells.The yield of CFU-S (colony-forming unit-spleen) was also analyzed by transplanting ES cell derivatives into lethally irradiated mice.[Results] Expression of Flk1,Integrin α4,Sca-1,and CD34 could be tested on induced EB cells.The percentage of Flk-1+,Integrin α4+ and Sca-1+ cells induced by were 3.03%,2.9%,and 13.74%,respectively,which are greater than other groups.The percentage of CD34+ cells induced by BMSC-CM was 1.07% which was greater than other groups.The yields of HPP-CFC from hematopoietic cells induced by FLSC-CM or BMSC-CM were 7.4 /105 cells (P ＜ 0.01) and 5.8 /105 cells (P ＜ 0.05) which were greater than the yields of control group.The yields of CFU-S from hematopoietic cells induced by FLSC-CM or BMSC-CM were 8.5/5 × 105 cells and 6.75/5 × 105 cells which were also greater than the yields of control group (P ＜ 0.001).[Conclusion] Both YS-SCCM,FL-SCCM,and BM-SCCM could promote hematopoietic differentiation of ESE14.1 cells.Hematopoietic differentiation induced by FL-SCCM or BM-SCCM is more effective,which generates hematopoietic progenitor cells with normal function.Application of FL-SCCM generates more primitive hematopoietic progenitor cells than that of BM-SCCM.

The purpose of this study is to investigate the preparation of hydroxycamptothecine (HCPT)-loaded cubic crystal liquid embolic precursor solution, and evaluate its in vitro embolic efficiency. Phytantriol was used as cubic crystal liquid embolic material, and the optimal formulation was selected according to ternary phase diagram. Polarized light microscopy, differential scanning calorimetry, and small angle X-ray scattering (SAXS) were used to characterize the cubic crystal structure. High performance liquid chromatography and X-ray diffraction analysis were used to investigate the lactone ring of HCPT. In vitro dissolution was preliminary evaluated, and the simulation embolic model was constructed to evaluate the embolic efficiency of precursor solution. Meanwhile, the gelation time and adhesion force were investigated. The results showed that HCPT-loaded precursor solution for embolization had been successfully prepared with low viscosity which was injectable. The precursor solution could transform into Pn3m structure liquid crystal phase gel rapidly when contracting with excess water. The formed HPCT gel remained its lactone form as the same in precursor solution, and expressed the good ability to block the saline flow, and HCPT could keep sustained releasing drug over 30 days. The prepared drug-loaded embolic precursor solution showed a promising potential for vascular embolization and application in clinical treatment of tumor.
Antineoplastic Agents, Phytogenic ; chemistry ; Calorimetry, Differential Scanning ; Camptothecin ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; chemistry ; Fatty Alcohols ; chemistry ; Liquid Crystals ; Scattering, Small Angle ; Water ; X-Ray Diffraction

OBJECTIVETo observe the expression change of signal regulatory protein alpha1 (SIRPalpha1) in autoimmune hepatitis (AIH) and approach the relationship between SIRPalpha1 and the extent of inflammation.

METHODSImmunohistochemistry is used to detect the expression of SIRPalpha1 in the paraffin section preparations of 33 AIH and 10 normal hepatic tissue.

RESULTSSIRPalpha1 is positive or weakly positive expressed in AIH. The staining is localized in the cytoplasm of Kupffer cells in the hepatic sinusoid with focal distribution. It is negative in normal hepatic tissue. In light AIH, it is negative or weakly positive expressed with a 36.4 percent of the positive rate (4/11). The positive or strong positive expression is found in the moderate AIH with an 84.2 percent of the positive rate(16/19). There is statistical significance between both light AIH, moderate AIH and severe AIH (P less than 0.001) and moderate AIH and light AIH (P less than 0.001). There is no statistical significance between both light AIH and severe AIH (P = 0.145 ) and moderate AIH and severe AIH (P = 0.084).

CONCLUSIONSAs a negative regulatory factor, the expression of SIRPalpha1 in hepatic sinusoid Kupffer cells is some associated with the extent of AIH.

Adolescent ; Adult ; Aged ; Antigens, Differentiation ; metabolism ; Cell Communication ; Child ; Female ; Hepatitis, Autoimmune ; metabolism ; pathology ; Hepatocytes ; metabolism ; pathology ; Humans ; Kupffer Cells ; metabolism ; pathology ; Male ; Middle Aged ; Receptors, Immunologic ; metabolism ; Young Adult

Objective To investigate the prevalence, common clinical symptoms and complications, transmission routes and media of brucellosis among human in the city of Songyuan in Jilin province, and to provide practical basis for brucellosis intervention and related control measures. Methods Use self-designed questionnaire to collect information from outpatients in brucellosis clinic in Songyuan Center for Disease Control and Prevention from January to June 2009, and to analyze the related data from the survey: prevalence, time and geographical distribution, clinical symptom, transmission route and media. Results Of the total 620 cases investigated, there were 284 patients accounting for 45.8% (284/620), 75 suspected patients accounting for 12.1% (75/620), 13stealth patients accounting for 2.1% (13/620) and 248 negative people accounting for 40.0% (248/640). Main common symptoms of the patients were fever[66.5%( 189/284)], muscle and joint pain[38.7%( 110/284)],fatigue[27.5%(78/284)], hyperhidrosis[25.0%(71/284)]and low back pain[17.3%(49/284)]. The patients group had a significantly higher prevalence of mucocutaneous infection, contacting infected animal abortion flow,fur, soil, faeces and dust than the uninfected group(χ2 value were 27.12, 22.75, 8.90, 6.65, 6.39, 6.39, all P< 0.01 or < 0.05). Conclusions The positive rate of brucellosis in the brucellosis clinic of Songyuan city is high,and patients have typical symptoms. We should take comprehensive control measures to protect the high-risk group and reduce the local infectivety.

This paper was aimed to investigate the effects of ATP-sensitive potassium channels on the proliferation and differentiation of rat preadipocytes. We examined the expression of sulphonylurea receptor 2 (SUR2) mRNA in preadipocytes and adipocytes obtained by inducing for 5 d and the effects of the inhibitor (glibenclamide) and opener (diazoxide) of ATP-sensitive potassium channels on the expression of SUR2 mRNA in preadipocytes by real-time PCR. Preadipocyte proliferation and cell cycle were measured by MTT spectrophotometry and flow cytometer. The content of intracellular lipid was measured by oil red O staining, cell diameter was determined by Image-Pro Plus 5.0 software and the expression of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) mRNA was estimated by RT-PCR. SUR2 mRNA was expressed in both preadipocytes and adipocytes obtained by inducing for 5 d, and the expression in adipocytes was obviously higher than that in preadipocytes. Glibenclamide inhibited the expression of SUR2 mRNA in preadipocyte, promoted preadipocyte proliferation in a dose-dependent manner, increased the cell percentages in G(2)/M + S phase, increased lipid content, augmented adipocyte diameter, and promoted the expression of PPAR-gamma mRNA. But the actions of diazoxide were contrary to those of glibenclamide. These results suggest that ATP-sensitive potassium channels regulate the proliferation and differentiation of preadipocytes, and PPAR-gamma is probably involved in the effect of ATP-sensitive potassium channels.
ATP-Binding Cassette Transporters ; genetics ; metabolism ; Adipocytes ; cytology ; Animals ; Cell Differentiation ; physiology ; Cell Proliferation ; Cells, Cultured ; KATP Channels ; physiology ; Male ; Obesity ; pathology ; PPAR gamma ; metabolism ; Potassium Channels, Inwardly Rectifying ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Drug ; genetics ; metabolism ; Sulfonylurea Receptors

OBJECTIVETo identify the enhancers of human lung specific X protein (LUNX) and their regulation at the transcription level in vitro.

METHODSThree enhancer fragments (E1:+3770~+3959bp; E2: +6454~+6555bp; E3: +14553~+14652 bp) predicted by bioinformatics software were isolated from the human genomic DNA by PCR amplification. Luciferase assay was performed to detect the activities of the enhancers in transcriptional regulation.

RESULTSPCR products were confirmed by DNA sequencing. The amplified enhancers digested by Kpn I/Xho I and BamH I/Sal I, to generate the sticky-end fragments were inserted into PGL3-promoter in a reporter vector, and 6 luciferase expression vectors were obtained. All the reporter plasmids and pGL3-promoter were transiently transfected into HEK293 cells with an internal control of pSV-β-Galactosidase reporter vector. The enhancer activity of each construct was evaluated by luciferase assay of the cell extracts after transfection for 48 h. The results showed that the 3 fragments, when located upstream, did not increase transcription of reporter gene, but when at the downstream, E1 and E3 increased the transcription by 2.83 and 1.59 folds of that of pGL3-promoter, respectively.

CONCLUSIONLUNX gene sequences from +3770 to +3959 bp and +14553 to +14652 bp possess the capacity to enhance gene transcription.

Base Sequence ; Cloning, Molecular ; Enhancer Elements, Genetic ; Gene Expression Regulation ; Glycoproteins ; genetics ; HEK293 Cells ; Humans ; Molecular Sequence Data ; Phosphoproteins ; genetics ; Transcription, Genetic

OBJECTIVETo estimate the prevalence of attention deficit hyperactivity disorder (ADHD) in children with epilepsy, and the factors that may contribute to the prevalence of co-morbidity between ADHD and epilepsy.

METHODSA total of 256 children aged 6-15 years old who were diagnosed with epilepsy were enrolled. The prevalence of ADHD in children with epilepsy, and the factors that may contribute to the development of co-morbidity between ADHD and epilepsy were explored.

RESULTSThe systematic evaluation in 192 patients was completed. Of the 192 children, 81 (42.2%) were diagnosed with ADHD. The earlier the epilepsy onset, the higher the frequency of the co-morbidity of ADHD occurring. The longer the period of antiepileptic medication, the higher the prevalence of the co-morbidity of ADHD. Epileptic children receiving a combination of antiepileptic drugs had a higher prevalence of ADHD. ADHD was more common in children with some specific types of epilepsy, such as Lannox-Gastaut syndrome and generalized tonic-clonic epilepsy, or epilepsy with multifocal epileptic discharges in the EEG record.

CONCLUSIONSADHD occurs frequently in children with epilepsy. The factors associated with increased risk of ADHD include the onset age of epilepsy, the types of seizures or epileptic syndromes, the epileptiform EEG discharges, and the effects of antiepileptic drugs.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; epidemiology ; etiology ; Child ; Comorbidity ; Electroencephalography ; Epilepsy ; complications ; drug therapy ; physiopathology ; Female ; Humans ; Male ; Prevalence