A Novel Gene Mutation of Runx2 in Cleidocranial Dysplasia / 华中科技大学学报（医学）（英德文版）

Haploinsuffieiency of the runt-related transcription factor 2 (Runx2) gene is widely known to be responsible for cleidocranial dysplasia (CCD).To date,more than 190 mutations in Runx2 gene have been reported to be related to CCD.In this study,a novel mutation of Runx2 gene was observed in a female with CCD.Genomic DNA was extracted from peripheral venous blood of the proband and eleven members of her family.Genetic testing on these twelve people identified a novel missense mutation (c.895T＞C,Y299H) in exon 5 of the RUNX2 gene in the proband.This mutation results in an amino acid change at codon 895 (P.Tyr 299 His.) from a tryptophan codon (TAT) to a histidine codon (CAT).Our finding may further extend the known mutation spectrum of the RUNX2 gene,and facilitate prenatal genetic diagnosis of CCD in the future.

Effects of exogenous H2S on H2S concentration and cystathionine β-synthase expression in hippocampus in a rat model of vascular dementia / 国际脑血管病杂志

Objective To investigate the effect of exogenous hydrogen sulfide (H 2 S) on H2 S concentration and cystathionine β-synthase (CBS) expression in hippocampus in a rat model of vascular dementia (VaD). Methods A rat model of VaD was induced by using the modified four -vessel occlusion. The rats were divided into sham operation, model, low -dose and high-dose NaHS groups using the random number table method. They were further redivided into one day, seven -day, and 30-day subgroups according to the time after modeling. After modeling respectively, NaHS 30 μmol/kg and 100 μmol/kg were injected intraperitoneally every day in the low -dose and high-dose NaHS groups. The normal saline was injected intraperitoneally every day in the sham operation group and the VaD model group. Morris water maze test was used to evaluate the learning and memory ability of the rats. The expression of CBS in hippocampus was detected by real-time fluorescent polymerase chain reaction. Western Blotting was used to detect expression of CBS protein in hippocampus. Results Morris water maze test showed that the escape latencies of the model group, low -dose and high-dose NaHS groups were prolonged significantly compared with the sham operation group (P <0.05); the times of crossing the platform were decreased significantly compared with the model group (P <0.05); and the escape latencies were shortened significantly in the low -dose and high-dose NaHS groups compared with the model group ( P <0.05). The H2 S content in hippocampus was decreased significantly in the model group, low -dose and high-dose NaHS groups compared with the sham operation group, but the low -dose and high-dose NaHS group was significantly higher than that in the model group (all P <0.05). The expression of CBS mRNA and protein in the model, low -dose and high-dose NaHS groups was significantly lower than that of the sham operation group (all P < 0.05), and there was no significant difference between the low -dose and high-dose NaHS groups and the model group. Conclusions Exogenous H2 S may improve the learning and memory ability of the VaD rats. It may be associated with the increased H2 S content in hippocampus. However, it has no effect on CBS expression.

Molecular diagnosis for a novel deletion mutation of α thalassemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To raise awareness of the pathogenesis and diagnosis of thalassemia by reporting one case of α thalassemia patient with a large deletion fragment and analyzing the pedigree.</p><p><b>METHODS</b>Firstly, blood cells and hemoglobin electrophoresis analysis were used for screening of thalassemia, and then three common kinds of deletional α thalassemia in Chinese was detected by Gap-PCR, three common kinds of non- deletional α thalassemia and seventeen common mutations of β thalassemia in Chinese were analyzed by using PCR- RDB. The unknown mutation of samples was identified with Multiplex Ligation-dependent Probe Amplification (MLPA) and DNA sequencing.</p><p><b>RESULTS</b>The proband female presented with microcytic hypochromic anemia(hemoglobin 71 g/L, mean corpuscular volume 52.4 fl, mean corpuscular hemoglobin 16.1 pg), and hemoglobin A2 1.4%. The identified large deletion fragment length was 21 925 bp, so far which had not been reported in the world and was named -α²¹·⁹. It was registered in USA DNA database and GenBank accession number as KF360979. The genotype of her mother and father and brother were αα/-α²¹·⁹, --(SEA)/-α³·⁷, αα/-α³·⁷ respectively, and the genotype of her and her sister were the same of --(SEA)/-α²¹·⁹. Her husband gene of thalassemia had no mutation, so prenatal diagnosis of thalassemia was not carried out in the pregnant woman.</p><p><b>CONCLUSION</b>The discovery of -α(21.9) deletion mutation was enriched the DNA mutation gene database of thalassemia, and had important significance for genetic counseling and thalassemia prenatal diagnosis.</p>

Survival of engineered myocardium constructed by bone marrow mesenchymal stem cells in vitro / 中国组织工程研究

BACKGROUND:So far, engineered myocardium is stil facing many problems. Research has demonstrated that bone marrow mesenchymal stem cells can be differentiated into myocardial cells. Polyglycolide and polycaprolactone are commonly used artificial polymers, which have good biocompatibility. OBJECTIVE:To observe the growth of the poly(glycolic acid)/poly(e-caprolactone) copolymer patch in vitro in normal myocardium and infarcted myocardium. METHODS:Bone marrow mesenchymal stem cells in Sprague-Dawley rats were separated using adherent separation and selection method, cultured in vitro. The third passage was labeled with DAPI. The bone marrow mesenchymal stem cellsuspensions (2×106/cm2) were produced and planted on poly(glycolic acid)/poly(e-caprolactone) copolymer scaffolds to form poly(glycolic acid)/poly(e-caprolactone) copolymer patch. After culturing for 48 hours, the specimens were observed under electron microscope, stained with hematoxylin and eosin, and then observed under light microscope. Rat models of myocardial infarction were established by ligating left anterior descending coronary artery. Poly(glycolic acid)/poly(e-caprolactone) copolymer patch was implanted into the normal and infarcted myocardium for 5 weeks. The survival of bone marrow mesenchymal stem cells was determined by the detection of pathology. RESULTS AND CONCLUSION:Results of light microscope and electron microscope demonstrated that bone marrow mesenchymal stem cells grew three-dimensional y on poly(glycolic acid)/poly(e-caprolactone) copolymer patch. cells and patch were adhesive wel . Under laser confocal microscopy, compared with the first week, bone marrow mesenchymal stem cells were marked by DAPI in the myocardium at the fifth week. There were bone marrow mesenchymal stem cells marked by DAPI in the infracted area. Results of hematoxylin-eosin staining exhibited that bone marrow mesenchymal stem cells were detected in the infarct area. These results suggested that bone marrow mesenchymal stem cells adhered to the poly(glycolic acid)/poly(e-caprolactone) copolymer stent wel . The complexes of poly(glycolic acid)/poly(e-caprolactone) copolymer and bone marrow mesenchymal stem cells can be used for reparation of myocardium.

Determination of the diameters of fetal umbilical artery and umbilical vein in 12 - 28 gestational weeks in screening thalassemia with ultrasound / 中华超声影像学杂志

Objective To study the value of the diameters of fetal umbilical artery and umbilical vein in early prenatal screening program for thalassemia by ultrasound.Methods A total of 120 cases of singleton pregnant with thalassemia at 12- 28 gestational weeks were detected by ultrasound,328 cases of normal singleton pregnant at 12 - 28 gestational weeks were selected as controls.The diameters of fetal umbilical artery and umbilical vein were tested and compared between the two groups.The diagnostic values were evaluated by ROC curve.Results At 12 - 15+6 gestational weeks,the diameters of umbilical artery and umbilical vein of thalassemia group were larger than those of control group(P ＜0.05).At 16 - 19+6 gestational weeks, the diameter of umbilical vein of thalassemia group was larger than that of control group,the differences were significant (P ＜0.05).The cutoff threshold of the diameter of fetal umbilical artery was 1.80 mm,the diameter of fetal umbilical vein was 2.95 mm at 12 - 15+6 gestational weeks,the diameter of fetal umbilical vein was 3.80 mm at 16- 19+6 gestational weeks,which were obtained by ROC analysis.The sensitivity of this cutoff threshold to identify thalassemia were 83.3％, 83.3％, 47.8％,respectively, while the specificity were 90.5 ％, 95.2 ％, 85.4％, respectively.Conclusions The diameters of fetal umbilical artery and umbilical vein detected by ultrasound have great value on early prenatal screening of thalassemia, these indexes can be used as an important factors in early prenatal screening of thalassemia.

Bioconversion of apple pomace into a multienzyme bio-feed by two mixed strains of Aspergillus niger in solid state fermentation

Apple pomace is a wasted resource produced in large quantities and its deposit has caused serious environmental problems, so it is significance to make the full utilization of the residues. The objectives of this work were to produce multienzyme bio-feed, biodegrade the anti-nutritional factors such as pectin and tannins in apple pomace, and obtain the nutritional enrichment of the fermented substrate. The mixture of apple pomace and cottonseed powder (1:1, w/w), supplemented with 1 percent (w/w) (NH4)2SO4 and 0.1 percent (w/w) KH2PO4, was proved to be the optimum medium for the mixed strains of Aspergillus niger M2 and M3 (2:1, w/w). The activities of pectinase, proteinase and cellulase achieved 21168 u/g, 3585 u/g and 1208u/g, and the biodegradation rates of pectin and tannins reached 99.0 percent and 66.1 percent, respectively, when 0.4 percent(w/w) of the test fungiwere inoculated and incubated at 30ºC for 48 hrs in solid state fermentation. The utilization of apple pomace in the paper can be served as a model for the similar waste recycling.

Determination of the cysteine residues in the surface-confined biomolecules by using electrochemical desorption and fluorescence detection / 药学学报

To develop a method for the detection of surface-confined peptides containing cysteine residues or oligodeoxynucleotides (ODNs) whose 3' ends modified with thiol groups, and a thiol-specific fluorescent cross-linker, N-(9-acridinyl) maleimide (NAM) was used. The peptides studied herein include both the oxidized and reduced forms of glutathione, and a hexapeptide (FT). Peptides are first attached onto the activated 11-mercaptoundecanoic acid (MUA)-terminated alkanethiol self-assembled monolayers (SAMs) and then derivatized with NAM. The cysteine residues was determined by using electrochemical desorption and fluorescence detection. GSH concentration as low as 40 pmol x L(-1) can be measured. The fluorescence intensity in the case of FT is about 3 times as high as that for GSH, which is consistent with the molar ratio of cysteine residues in these two molecules. The analytical performance of gene analysis was also evaluated through the analyses of a complementary target and targets with varying numbers of mismatching bases. The method described here is simple, sensitive, reproducible, and does not require sophisticated analytical instrumentation and separation procedures.

Polarographic behavior and determination of glimepiride / 药学学报

<p><b>AIM</b>To establish a polarographic method of parallel catalytic hydrogen wave for determination of glimepiride.</p><p><b>METHODS</b>The catalytic wave of glimepiride in the presence of K2S2O8 was used to improve the analytical sensitivity. The rapid determination of glimepiride was done by linear single sweep polarography.</p><p><b>RESULTS</b>The catalytic hydrogen wave of glimepiride was measured at ca. -1.36 (vs SCE) in 0.09 mol x L(-1) Na2B4O7-KH2PO4 (pH 6.24 +/- 0.1) supporting electrolyte. When 1.0 x 10(-2) mol x L(-1) K2S2O8 was present, the current increased by 25 times, and the peak potentioal was unchanged, producing a more sensitive parallel catalytic hydrogen wave. The peak current of the parallel catalytic hydrogen wave was rectilinear to the glimepiride concentration in the range 1.0 x 10(-7) - 4.2 x 10(-5) mol x L(-1) (r = 0.9990, n = 9). The detection limit was 5.0 x 10(-8) mol x L(-1).</p><p><b>CONCLUSION</b>The proposed method could be applied to the determination of glimepiride in pharmaceuticals without preliminary separation.</p>

Polarographic catalytic wave of clarithromycin and its applications / 药学学报

<p><b>AIM</b>To develop a new method for the determination of clarithromycin.</p><p><b>METHODS</b>The catalytic wave of clarithromycin in the presence of K2S2O8 was used for improving the analytical sensitivity. The rapid determination of clarithromycin has been carried out by linear single sweep polarography.</p><p><b>RESULTS</b>The reduction wave of clarithromycin appeared at ca. -0.79 V (vs SCE) in 0.24 mol x L(-1) KH2PO4-Na2HPO4 (pH 6.81) supporting electrolyte, which was ascribed to the reduction of carbonyl group on C-9 position. In the presence of 0.01 mol x L(-1) K2S2O8, the reduction wave was catalyzed to produce a parallel catalytic wave. The peak current of the catalytic wave was ca. Twenty times higher than that of the corresponding reduction wave. Based on the catalytic wave, a new method for the determination of clarithromycin has been proposed. The peak current of the catalytic wave was rectilinear to clarithromycin concentration in the range of 4.0 x 10(-7)-5.0 x 10(-5) mol x L(-1). The detection limit was 2.0 x 10(-7) mol x L(-1).</p><p><b>CONCLUSION</b>The proposed method could be used for the direct determination of clarithromycin in pharmaceuticals and urine without preliminary separation.</p>