Objective To explore the clinical effect of single use or combination of dydrogesterone and progestin in treatment of threatened abortion caused by uteal phase defect.Methods Totally 186 patients with threatened abortion caused by uteal phase defect accepted in The First Affiliated Hospital of Henan from April 2015 to April 2016 were selected and randomly divided into groups A,B,and C with 62 cases in each group.Patients in group A were given dydrogesterone,those in group B were given progestin,and those in group C were given dydrogesterone combined with progestin.Then the clinical effect,expression of hormones,treatment outcome,and adverse reaction were observed and compared.Results The total effective rates of groups A and B were 72.58％ and 66.13％,respectively,which were obviously lower than 90.32％ of group C with statistically significance (P ＜0.05).The expression levels of P,E2,and hCG of three groups after treatment were higher than those before,those in group C were the highest among them (P ＜ 0.05).The successful treatment rates of groups A,B,and C were 83.87％,82.26％,and 95.16％,respectively,which had no great difference.Conclusion Combination use of dydrogesterone and progestin has better effective rate in treatment of threatened abortion caused by uteal phase defect compared to single use of these two drugs,which has good safety and worth of clinical application.

OBJECTIVE: To synthesize and characterize dapsone-alginate acid (DS-ALG) conjugate. METHODS: Alginate (Alg) was selected as the drug carrier and valine (Val) as the linking arm to synthesize DS-Alg, which could be applied to topical administration. And the synthetic condition of DS-Alg was optimized by changing the amount of 1-(3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), while the pH of solvent was changed in the range of 4.0 to 6.0. The structures of the products were characterized by 1H-NMR, MS and FT-IR. Meanwhile, the drug release in vitro of DS-Alg was investigated in the mixture of pH 7.4, 0.05 mol•L -1 PBS and ethanol by diffusion cells. The concentration of DS or valine-dapsone in the release medium was detected by HPLC. Taking rats with local scald as model, the drug release in vivo was measured by coating the trauma with DS-Alg conjugate cream and monitoring the drug concentration in blood. RESULTS: The optimum synthetic conditions of DS-Alg were as follows: 0.277 g valine-dapsone, 0.400 g sodium alginate, 7 eq EDC, 3 eq NHS, pH of the solvent of 5.5. And during 72 h, there was no DS detected in the release medium or rat plasma. The AUC0→72 h of DS-Alg was 0. It suggested that DS immobilized by Alg with covalent bond was too stable to be released from Alg in vitro and in vivo. The DS-Alg conjugate could effectively prevent DS from entering the systemic circulation. CONCLUSION: DS-Alg conjugate is successfully synthesized. The conjugate is stable that DS cannot be released from the conjugate to the bloodstream, which can efficiently decrease the side effects of DS and has the potential for topical administration.

OBJECTIVETo study the flavanoids extracted from onion on the blood-brain barrier (BBB) permeation, and their effects on primary cultured neuron cell proliferation and apoptosis of SD rats using ethanol reflux method.

METHODSThe brain microvascular endothelial cells (BMVECs) were first successfully primary cultured. Then rats BMVECs and astrocytes (ACs) were co-cultured to establish the in vitro BBB model. The flavanoids were extracted from onion using ethanol reflux method. The model was verified by transmission electron microscopy (TEM) and trans-epithelial electric resistance (TEER). The flavanoids permeability was tested using high performance liquid chromatography (HPLC). Meanwhile, rat neuron cells were cultured and exposed to H2O2 and flavanoids. Their effects on the cell proliferation and apoptosis were observed using MTT assay. The injury of neuron DNA was analyzed using single-cell gel electrophoresis (SCGE) and immunofluorescent assay.

RESULTSThe in vitro BBB model was successfully established by TEM and TEER. Results of HPLC proved flavanoids extracts could effectively permeate the BBB with the permeability of 60.58%. The extractive at 10 - 20 microg/mL showed obvious inhibition on the apoptosis of neuron cells induced by H2O2, and attenuated the injury of neuron DNA.

CONCLUSIONSThe flavanoids extracted from onion ethanol reflux method could effectively penetrate the BBB. They also showed obvious inhibition on the H2O2 induced neuron cell apoptosis and DNA injury.

Animals ; Animals, Newborn ; Apoptosis ; Astrocytes ; cytology ; drug effects ; Blood-Brain Barrier ; drug effects ; Cells, Cultured ; DNA Damage ; Endothelial Cells ; cytology ; drug effects ; Flavonoids ; pharmacology ; Hydrogen Peroxide ; Neurons ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; Onions ; chemistry ; Rats

OBJECTIVETo determine the potential urinary biomarkers of metabolic syndrome (MS) with early renal injury and establish diagnostic models by magnetic bead-based separation and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

METHODSParticipants were selected from the epidemiologic study on MS and renal involvement among residents in Pinggu district, Beijing. Eight-hour overnight urine samples were fractionated by means of magnetic bead-based weak cation exchange chromatography and subsequently analyzed with MALDI-TOF-MS. Wilcoxon test and random forests were used to screen differential protein peaks of MS patients with early renal injury, then combined with genetic algorithm and support vector machine, respectively, to establish diagnostic models.

RESULTSTotally 54 cases of MS without renal injury and 46 cases of MS with early renal injury were enrolled. Totally twenty protein peaks were up-regulated in the urine of MS patients with early renal injury by Wilcoxon test (P < 0.05); random forests algorithm revealed twelve protein peaks up-regulated in the urine of MS patients with early renal injury (importance value of mean decrease in accuracy > 0.005). Genetic algorithm based model showed 82.6% sensitivity, 84.3% specificity, and 83.5% accuracy by a 10-fold cross-validation in identifying MS patients with early renal injury; correspondingly, the support vector machine based model reported 89.2% sensitivity, 81.1% specificity and 85.5% accuracy. Four protein peaks were included in two diagnostic models with mass-to-charge ratios of 2756.98, 3019.11, 9077.04, and 10 054.26.

CONCLUSIONSThe urinary proteome patterns of MS with early renal injury were successfully established with magnetic bead-based separation and MALDI-TOF-MS technology. A series of urinary differential expressing protein peaks were identified with bioinformatics tools. Diagnostic models combining cluster of protein peaks are capable of differentiating MS patients with early renal injury from those without renal injury. The different urine protein excretion patterns revealed in this study provide urinary candidate biomarkers of MS patients with early renal injury for future identification and biological roles investigation.

Adult ; Biomarkers ; urine ; Chromatography, Ion Exchange ; methods ; Female ; Humans ; Kidney Diseases ; etiology ; urine ; Male ; Metabolic Syndrome ; complications ; urine ; Middle Aged ; Proteome ; analysis ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Urine ; chemistry

OBJECTIVETo investigate the molecular genetic mechanism of a Chinese patient with mucopolysaccharidosis type I (MPS I).

METHODSPCR-sequencing analysis was applied to detect the mutations in exons in alpha-L-iduronidase gene (IDUA) of the patient. Restriction fragment length polymorphism (RFLP) and allele-specific oligonucleotide hybridization (ASO) were used to confirm the identified mutations. PCR amplified DNA samples from 50 normal individuals were sequenced to demonstrate that the newly identified mutation was not polymorphism.

RESULTSThe patient was compound heterozygous for a previously reported nonsense mutation Q60X (178C > T) in exon 2, inherited from the mother, and a newly detected missense mutation D203N (607G > A) in exon 6 from the father. The newly identified mutation D203N was not found in PCR amplified products from 50 normal individuals, indicating that it was not polymorphism.

CONCLUSIONThe two identified mutations may be the cause resulting in patient's clinical phenotype.

Adolescent ; Base Sequence ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Humans ; Iduronidase ; genetics ; Male ; Mucopolysaccharidosis I ; genetics ; Mutation ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism

OBJECTIVETo investigate clinical and molecule genetics features of four Ph-positive leukemia patients characterized by pericentric inv(9)(p22q34) with the der(9)t(9;22)(q34;q11).

METHODSCytogenetic analysis was carried out on bone marrow directly or after short-period culture. R banding was used for karyotype analysis. BCR/ABL fusion gene was detected with interphase fluorescence in situ hybridization (FISH), and chromosome painting was carried out using specific probes. RT-PCR was used to detect BCR/ABL chimeric transcripts.

RESULTSOne patient with acute myeloid leukemia (AML) presented three clones, which included one with a normal karyotype, one with t(9;22)(q34;q11), and one with inv(9)(p22q34) involving the der(9)t(9;22) and additional t(8;12)(q12;p11). The inv(9)(p22q34) has always co-occurred with der(9)t(9;22)(q34;q11) accompanied by der(22)t(9;22)(q34;q11) in all metaphases from the three patients with chronic myeloid leukemia (CML). B3a2 transcript was detected in all patients by RT-PCR. Inv(9)(p22q34) was found in both CML and AML, and was associated with poor prognosis.

CONCLUSIONInv(9)(p22q34) is a novel, rare, but recurrent secondary chromosomal abnormality for Ph-positive leukemia. Leukemia with der(9)t(9;22) and inv(9)(p22q34) has unique clinical and laboratory characteristics.

Adult ; Chromosome Inversion ; Chromosomes, Human, Pair 22 ; Chromosomes, Human, Pair 9 ; Female ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Translocation, Genetic

OBJECTIVETo identify suitable hydroxyl polycyclic aromatic hydrocarbons (OH-PAHs) for co-evaluation of internal exposure level of PAHs by simultaneous determination of a variety of OH-PAHs in urine.

METHODSThe 24-h individual particulate matter and morning urine samples of 112 subjects were collected during June 2011. PAHs carried by individual particulate matter samples and OH-PAHs in urine samples were detected by gas chromatography-mass spectrometry.

RESULTSSeven OH-PAHs were detected in urine samples, among which 1-hydroxy-naphthalene (1-OHNap) concentration was the highest [(20.54 ± 28.94) µmol/mol Cr], while 1-hydroxy-pyrene (1-OHP) concentration was the lowest [(0.73 ± 0.63) µmol/mol Cr]. The concentrations of these seven OH-PAHs decreased in the following order: 1-hydroxy-naphthalene (1-OHNap) > 9-hydroxy-fluorene (9-OHFlu) > 2-hydroxy-naphthalene (2-OHNap) > 3-hydroxy-fluorene (3-OHFlu) > 2-hydroxy-fluorene (2-OHFlu) > 6-hydroxy-chrysene (6-OHChr) > 1-hydroxy-pyrene (1-OHP). The effects of gender and smoking upon the contents of OH-PAHs in urine samples were not significant. There was a good correlation between total hydroxy-naphthalene (ΣOHNap) and 1-OHNap (r = 0.948), and a good correlation was also showed between total hydroxy-fluorene (ΣOHFlu) and 9-OHFlu (r = 0.975). Naphthalene carried by atmospheric particulate matters demonstrated better correlation with 1-OHNap than 2-OHNap, while fluorene carried by atmospheric particulate matters showed better correlation with 9-OHFlu than 3-OHFlu and 2-OHFlu. The correlation coefficients of ΣOHNap, ΣOHFlu and 6-OHChr with 1-OHP were 0.427, 0.543 and 0.655, respectively, and the correlations were not strong.

CONCLUSIONIt cannot reflect internal exposure level of PAHs to use 1-OHP as the only biomarker, while 1-OHNap and 9-OHFlu can be well predictive of the exposure levels of corresponding total OH-PAHs, suggesting that simultaneous determination of 1-OHNap, 9-OHFlu and 1-OHP can be more accurate and comprehensive in evaluating the internal exposure level of PAHs.

Aged ; Air Pollutants ; analysis ; urine ; China ; Environmental Monitoring ; methods ; Female ; Humans ; Hydroxyl Radical ; analysis ; urine ; Male ; Middle Aged ; Polycyclic Aromatic Hydrocarbons ; analysis ; urine

Adenovirus transfection technique was used in the current study to show if thioredoxin-interacting protein (TXNIP) overexpression can induce cell apoptosis and injury in H9C2 cardiomyocytes cultured in normal glucose condition. And the mechanisms were then investigated. Briefly, H9C2 cardiomyocytes in logarithmic growth phase were randomly divided into three groups: normal cultured group, empty adenovirus vector group (Ad-eGFP) and TXNIP overexpression group (Ad-TXNIP-eGFP). All cells were cultured in DMEM containing normal concentration of glucose (5 mmol/L) and lipid. 72 h after adenovirus transfection, cells and culture mediums were collected for further assay. The results showed that Ad-eGFP and Ad-TXNIP-eGFP adenovirus transfected H9C2 cells successfully, and the transfection efficiency reached the peak at 72 h. Compared with Ad-eGFP group, Ad-TXNIP-eGFP transfection significantly increased TXNIP mRNA (P < 0.05) and protein expression level (P < 0.01). TXNIP overexpression induced remarkable cell apoptosis and injury as evidenced by increased caspase-3 activity (P < 0.05), apoptotic rate (P < 0.01) and LDH activity (P < 0.01). To further analysis the mechanisms of TXNIP-induced cell apoptosis, we also determined Trx activity, Trx related free radical injury and p38 kinase activation, which are involved in free radical induced apoptosis. The results showed that, compared with those in Ad-eGFP group, Trx activity was significantly decreased (P < 0.01), while malondialdehyde (MDA), 3-nitrotyrosine contents and p38 kinase activity were significantly increased (P < 0.01) in TXNIP overexpression group. These results suggest that TXNIP overexpression alone can induce severe apoptosis and injury in H9C2 cardiomyocytes even they are cultured in normal glucose and lipid concentration conditions. The mechanism involved is that overexpressed TXNIP can bind and inhibit Trx, impairs its antioxidative and antiapoptotic function, and then increases free radical induced injury and p38 kinase dependent apoptosis.
Adenoviridae ; genetics ; Animals ; Apoptosis ; Carrier Proteins ; genetics ; metabolism ; Caspase 3 ; metabolism ; Cell Line ; Genetic Vectors ; Myocytes, Cardiac ; cytology ; Rats ; Thioredoxins ; metabolism

OBJECTIVEThis study examined the effect of high concentration of phenylalanine (Phe) on Nogo-66 receptor (NgR) expression in the cortical neurons of rats in vitro in order to investigate whether NgR is involved in the etiology of Phe-induced brain damage.

METHODSNeurons from the cerebral cortex of embryonic rats were cultured for 3 days and then were treated with 0.9 mM Phe. After 12, 24 and 48 hrs of Phe treatment, mRNA and protein expression of NgR was detected by real-time PCR and Western blot respectively. Growth cones and growth axons of neurons were detected by immunofluorescence and immunohistochemistry respectively after 12 and 24 hrs of Phe treatment.

RESULTSThe length of growth axons of neurons was significantly shorter after 12 and 24 hrs of Phe treatment compared with the control group without Phe treatment (P<0.05). Growth cones collapse occurred in 12.5+/-9.7% and 24.1+/-4.5% of neurons respectively after 12 and 24 hrs of Phe treatment but only in 3.5+/-1.5% in the control group (P<0.01). The protein level of NgR after 12, 24 and 48 hrs of Phe treatment was up-regulated, with 9.0, 9.4 and 12.6 times as the control. mRNA level of NgR in the Phe treatment group did not differ from control.

CONCLUSIONSHigh concentration of Phe can induce an increased NgR protein expression in cortical neurons, and the increased NgR expression may contribute to the growth cones collapse and the inhibitory activities of axon regeneration after injury.

Animals ; Blotting, Western ; Cerebral Cortex ; chemistry ; drug effects ; GPI-Linked Proteins ; Immunohistochemistry ; Myelin Proteins ; analysis ; genetics ; Nogo Receptor 1 ; Phenylalanine ; pharmacology ; Polymerase Chain Reaction ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; analysis ; genetics