Allyamine or sodium vinyl sulfate was added into the solutions of acrylamide and N,N′-methylenebisacrylamide to make mixed monomer solutions. These solutions were respectively introduced in methacrylsiloxane-modifidied capillaries. In situ polymerization was initiated by thermally induced radicals. A polymer layer was chemically bonded to the inner wall, resulting in valently modified capillary columns. By altering the addition amounts of allyamine or sodium vinyl sulfate, differently charged polymeric layers were obtained. Five columns were prepared. The result indicated that columns modified with polymers containing amino groups were advantageous over those containing sulphonic acid groups or none. Further comparison revealed that the columns modified with a solution containing 0.15 mol/L acrylamide, 0.03 mol/L N,N′-methylenebisacrylamide and more than 1 mol/L allylamine exhibited good resolution. It is convenient to adjust electroosmotic flows(EOFs) in the columns by this way. In the meantime unspecific protein adsorption was efficiently depressed. When subjected) to zone electrophoresis of proteins from chicken egg white, the column showed high resolution and good reproducibility.

Objective To study the molecular genetic mechanism of a patient with 17? hydroxylase (CYP17) deficiency. Methods Genomic DNA were abstracted from the blood of the patient, her parents and healthy control. The 8 exons of CYP17 gene were amplified, using 5 pairs of designed primers, with polymerase chain reaction (PCR), and the 8 exons were sequenced by the dideoxy terminator method to determined the mutation sites. The corresponding exons of the parents of the patients were also amplified and sequenced to determine the zygosity of the patient and the source of the gene variances. Results The analysis revealed that the patient (46, XY) was a compound heterozygote carrying two different inherited mutations on CYP17 gene, one from mother containing a point mutation Arg 96 (C G G)→ Gln(C A G) and the other from father containing a nine base deletion (CACTCTTTC) at amino acid position 487～489 (Asp Ser Phe) near the carboxyl terminus of P450c17. Conclusion The CYP17 gene of the patient with 17? hydroxylase deficiency is a compound heterozygous mutation. The mutation changes the amino acid sequence of P450c17 enzyme, which in turn affected the enzymatic activity. Arg 96 is essential in P450c17 enzyme activity. Deletion of Asp 487 Ser 488 Phe 489 in exon 8 may be a prevalent mutation causing P450c17 deficiency in Southeast Asia.

Objective To investigate the prevalence and spectrum of β-thalassemia mutations in C, uangdong province, and provide a reference for prenatal diagnosis and genetic counseling in this population. Methods Three thousand two hundred and forty-seven blood samples were randomly selected from Guangzhou and 2984 blood samples from Shenzhen from January in 2005 to January in 2009. PCR and reverse dot blot hybridization (RDB) were adopted for detection of β-thalassemia mutations in Guangzhou and Shenzhen city. Results Seven hundred and fifty-one individuals in Guangzhou were found to have β-hemoglobin gene mutations, the detection rate was 23.13%(751/3247); 10 different mutations were identified, namely CD41-42(-TCTT), IVS-Ⅱ-654(C→T), -28(A→G), CDI7(A→T), CD71-72(+A), 13E, IVS-I-1(G→T), CD43(G→T), -29(A→G), CDI4-15(+G), which accounted for 42.53% (336/790) ,25.19% (199/790), 12.66% (100/790), 10.89% (86/790) ,3.29% (26/790), 2.15%(17/790), 1.27%( 10/790), 1.14%(9/790) ,0.51%(4/790) ,0.38%(3/790), respectively; the most common mutation was CD41-42(-TCTT), which accounted for 42.53%(336/790). In Shenzhen, 179 individuals were found to have β-thalassemia mutations, the detection rate was 6.00% (179/2984); 8 different mutations were identified excluding CD43 (G→T) and CD14-15 (+G); the most common mutation, however, was IVS-lI--654(C→T), which accounted for 40.44% (74/183). Conclusions The β-thalassemia mutations in Guangdong province are not only frequent, but also obviously heterogeneous, and the mutations differ from region to region. CD41-42 (-TCTT),ⅣS-Ⅱ-654(C→T), -28(A→G), CD17(A→T) were the 4 predominant mutations.

Objective To observe the role of dual-source CT (DSCT) in the diagnosis of coronary artery fistula. Methods Nine patients with coronary artery fistula were examined with dual-source CT coronary artery angiography. Then the source images were post processed using volume rendering (VR), multiple planar reformation (MPR), maximum intensity projection (MIP) and curved planar reformation (CPR), and sequential segmental analysis of the intracardiac and extracardiac anomalies was performed. Results Coronary artery fistula in all the 9 patients were accurately displayed with DSCT, including 2 with left main trunk to right ventricle fistula, 7 with left main trunk and (or) left anterior descending artery to main pulmonary trunk fistula, 5 with complex coronary-pulmonary artery fistula. Conclusion Dual-source CT coronary artery angiography is convenient, fast, non-invasive, and may be the preferable method for diagnosis of coronary artery fistula.

OBJECTIVE:To explore the effect of nifedipine combined with losartan on the blood pressure and renal function of hypertension complicated with coronary heart disease. METHODS:150 patients with hypertension complicated with coronary heart disease were randomly divided into observation group and control group. Control group was orally treated with Nifedipine sustained release tablet 30 mg,once a day;observation group was additionally treated with Losartan potassium capsules 50 mg,once a day. The treatment course for both groups was 1 month. Systolic blood pressure,diastolic blood pressure,renal function indicators be-fore and after treatment,and incidence of adverse reactions in 2 groups were observed. RESULTS:After treatment,systolic blood pressure and diastolic blood pressure after 3 months was lower than 1 month and lower than before treatment in same group,renal function indicators were significantly lower than before treatment,and observation group was lower than control group,the differ-ences were statistically significant(P<0.05);the incidence of adverse reactions in observation group was significantly lower than control group,the difference was statistically significant(P<0.05). CONCLUSIONS:Nifedipine combined with losartan can well control the blood pressure of hypertension complicated with coronary heart disease,protect renal functions,with good safety.

Background Compare with the lamellar laser refractive surgery,the laser slip on the surface operation is a better method in the correct of refractive error without the flap-related complications.However,whether to remain corneal epithelial flap and its influence for epithelial healing and visual restoration are still under investigation.Objective The aim of this study was to compare the short-term clinical efficacy of epipolis laser insitu keratomileusis (Epi-LASIK) and flap-free Epi-LASIK for correction of refractive error in high myopia.Methods The paired control design was adopted.Sixty eyes of consecutive 30 patients with high myopia were collected in this study.The written consent form was obtained from each patients prior to operation.Regular EpiLASIK was performed in the right eyes and the flap-free Epi-LASIK was carried out in the left eyes.Patients were followed-up for one month.The postoperative ocular irritation symptoms were scored,and the healing period of corneal epithelium and uncorrected visual acuity were examined at 1,3 and 5 days after operation.The naked visual acuity,diopter,best corrected visual acuity and the grading of haze were recorded and compared between the two groups at one month after operation.Results After surgery,the scores of corneal irritation were 3.13 ±0.43 and 3.30±0.47on the first day,showing insignificant differences between these two groups (t =- 1.54,P＞0.05 ).However,there existed statistically significant difference in the scores of corneal irritation on the third day after operation between the two groups (t =- 17.95,P＜0.01 ).The average time of corneal epithelial healing were (2.58 ±0.49 ) days in the flapfree Epi-LASIK group and ( 3.98 ±0.74) days in Epi-LASIK group with the significant difference between them (t=-11.36,P＜0.01 ).There was no statistical difference in uncorrected visual acuity in the first day (0.59±0.12 vs 0.58±0.11,t=0.39,P＞0.05),the fifth day (0.69±0.08 vs 0.67±0.09,t=0.84,P＞0.05) and 1 month (t =-1.90,P＞0.05 ).No significant difference was seen in the diopter between the two groups in 1 month after surgery (-1.03±0.45 vs -1.07 ±0.38,t =-0.31,P＞0.05 ).The eye numbers of corneal haze for 0 grade were 27 ( 90.00％ ) and over the 0.5 grade were 3 ( 10.00％ ) in flap-free Epi-LASIK group respectively,and those of corneal haze for 0 grade were 26(86.66％ ) and above the 0.5 grade were 4 (13.33％) in Epi-LASIK group respectively,showing a considerably difference between the two groups (x2 =0.0031,P＞0.05).Conclusions Both the flapfree or reserved epithelial Epi-LASIK allow the safe and effective correction of refractive error in high myopia,showing good predictability and stability in short term.Flap-free Epi-LASIK can relieve the corneal irritation symptom and accelerate the healing of corneal epithelium and recovery of visual acuity.

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

Background: Creatine transporter (CRTR) deficiency is the most common creatine deficiency syndrome, of which the final diagnosis relies on mutation in the X-linked CRTR gene. To date, more than 90 mutations in the SLC6A8 gene have been reported. This paper discusses a novel mutation detected via the thorough sequencing of all the X-chromosome-specific exons investigated in a four and a half year old boy with an intellectual disability, speech and language delay and motor disturbance. Methods: A brain magnetic resonance imaging (MRI) and a proton magnetic resonance spectroscopy (MRS) were carried out, the creatine and creatinine concentrations in the urine were checked and all exons were sequenced. Results: A detailed clinical investigation revealed a reduction in the cerebral creatine levels in the brain by the MRS, elevated creatine and creatinine concentrations in the urine and signal abnormalities in the left frontal cortex of the brain by the MRI. A novel change was identified in the heterozygosity of the exon 10: c.1395-c.1401 deletion. Conclusion: The use of a combination of powerful new technologies, such as thorough exome-nextgeneration sequencing and a brain MRS, should be considered, in order to determine any neurometabolic diseases, especially when the signal abnormalities in the brain MRI cannot be explained by any other factors. This mutation results most likely in a dysfunction of the creatine transport and synthesis, hence causing central nervous system symptoms.
Carrier Proteins

Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Endothelial Cells ; cytology ; drug effects ; physiology ; Female ; Humans ; Mice ; Mice, Inbred C57BL ; Neovascularization, Physiologic ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology

Objective: To screen the best genetic engineering bacterium for the production of peptide antibiotic hPAB-? and evaluate its fermentation level in bottle. Methods:After analysis of the interest fusion protein expression levels of 8 recombinant bacteria containing 1-8 copies of human peptide antibiotic hPAB-? expressing plasmid respectively,2-5 copies expressing bacteria were chosen for the further study of their bacteria yield,expression forms of the target protein, dissolution of the inclusion bodies and the efficiency of fusion protein purification by affinity chromatography, then the best engineering bacterium with the certain copies of interest peptide expressing plasmid was screened out and its optimal fermentation parameters in bottle were also studied. Results:The recombinant bacterium transformed by 3 copies of interest peptide expressing plasmid was the best candidate for its bacteria yield (3.153 g/L) and fusion protein expression level (27.7%) were the highest among 1-8 copies candidates. The inclusion bodies of 3 copies target fusion protein could be easily dissolved by 8 mol/L urea and captured by Ni-NTA column. The elution of the fusion protein could be directly cleaved to monomer by adding 2 mol/L hydroxylamine, adjusting pH to 9.0 and incubating at 45℃ for 2 h. The optimal fermentation conditions of the selected recombinant bacteria were: culture the organisms with modified M9-CAA media at 37℃ and 160 r/min to (A 600 )≈2.5, then add IPTG to the final concentration 100 ?mol/L to induce the expression of target fusion protein for 5 h. Conclusion:The engineering bacterium containing 3 copies interest peptide recombinant expressing plasmid is the best candidate for the production of peptide antibiotic hPAB-?,and its fermentation parameters are confirmed.