Objective To analyze the mutation distribution of phenylalanine hydroxylase(PAH)gene in pa-tients with phenylketonuria(PKU)in Ningxia,and understand the hot spots and regions of PAH gene. Methods First-ly,6 hot mutation exons including 3,5,6,7,11,12 and their surrounding introns of the PAH gene in 30 patients with PKU in Ningxia were directly sequenced. And then the last 7 exons of 1,2,4,8,9,10,13 were sequenced for the patients in which 2 mutant alleles were not characterized. Multiplex ligation - dependent probe amplification (MLPA)was performed for the identification of uncharacterized mutant alleles after PAH sequence analysis of patients with PKU. Results Among 60 alleles,there were 58 mutant alleles(96. 7% ). Forty - six(81. 6% )mutant alleles were found in the exons 3,5,6,7,11,12. A total of 23 various mutations were detected,including missense(n = 9), splicing(n = 9),nonsense(n = 2),small deletion(n = 2)and large deletion(n = 1). The most common mutations were R243Q(18. 3% ),IVS4 - 1G ﹥ A(11. 7% )and R111X(11. 7% ). Among them,a novel mutation N393del was detec-ted in exon 11. MLPA identified a large deletion(c. - 1932 + 3402del)in 3 patients,1 of them was homoallelic muta-tion,and the others were heteroallelic mutation. Conclusions There are obvious hotspots and hot spot areas of PAH gene in PKU patients in Ningxia. There is a large deletion mutation in PKU patients in Ningxia. The MLPA is an effec-tive assay to detect large deletion in PAH gene.

Objective To evaluate the PDCA cycle on the rehabilitation of postoperative patients with anal fistula, aiming at improving the recovery of patients and preventing recurrence. Methods 144 postoperative patients of anal fistula were randomly divided into the observation group and the control group with 72 cases in each group. The observation group received intervention based upon the PDCA cy-cle, and the control group was given the traditional oral propaganda. The recovery time, recurrence rate, mastering of education content and self-control ability were compared in the two groups using t test and χ2 test. Results The mastering level of education content and the self-management skills in the observation group were superior to those of the control group. After 3 to 24 months of follow-up, the observation group showed shorter recovery time and less recurrence rate than that of the control group. Conclusions Nurs-ing intervention based upon PDCA cycle can strengthen the patients' mastering of related knowledge, pro-mote the recovery, reduce recurrence rate and improve the patients' quality of life.

BACKGROUNDEpidemiological studies have shown that both active and passive cigarette smoking increase the risk of atherosclerosis. But very little is known about the biological processes induced by passive cigarette smoking that contribute to atherosclerosis. We observe the expression of a few of biological and inflammatory markers in human arterial walls in vitro which were treated with the second-hand smoke solution (sidestream whole, SSW), and discuss the possible mechanism of inflammatory injury induced by second-hand smoke.

METHODSThe biological markers (platelet endothelial cell adhesion molecule-1, PECAM-1; alpha-smooth muscle actin, alpha-SMA; collagen IV, Col IV) and inflammatory markers (vascular cell adhesion molecule-1, VCAM-1; monocyte chemoattractant protein-1, MCP-1; interleukin-8, IL-8) of human aortal wall were tested by immunofluorescence staining. The levels of MCP-1 and IL-8 mRNA expression were detected by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSNo distinct difference was observed between SSW and the control group on the expression of biological markers as assessed by the light microscope. But the inflammatory markers VCAM-1, MCP-1 and IL-8 on the subendothelial layer and smooth muscle cell layers, which are near the endothelium of arterial wall, were strongly stained in the SSW group compared with the control group. Their fluorescence intensities in the 1:40 SSW group (VCAM-1: 0.35 +/- 0.04, MCP-1: 0.34 +/- 0.05, IL-8: 0.37 +/- 0.05) and the 1:20 SSW group (VCAM-1: 0.40 +/- 0.04, MCP-1: 0.52 +/- 0.09, IL-8: 0.51 +/- 0.07) were significantly stronger than the control group (VCAM-1: 0.12 +/- 0.04, MCP-1: 0.06 +/- 0.02, IL-8: 0.24 +/- 0.03) by semi-quantitative analysis of immunofluorescence (P < 0.001 vs control). MCP-1 mRNA expression in the 1:40 SSW (0.15 +/- 0.04) and the 1:20 SSW (0.19 +/- 0.06) group was significantly higher than in the control group (0.09 +/- 0.03) (P < 0.05, P < 0.01 vs control); IL-8 mRNA expression in the 1:40 SSW (0.64 +/- 0.12) and 1:20 SSW (0.72 +/- 0.13) groups was also significantly higher than that in the control group (0.49 +/- 0.13) (P < 0.05, P < 0.01 vs control) by RT-PCR.

CONCLUSIONSIt is implied that a second-hand smoke solution induces the inflammatory reaction of the arterial wall by release of inflammatory factors even though there is no distinct structural change on the arterial walls under light microscope, indicating that passive cigarette smoking is related to inflammatory injury in human arterial wall and could be closely related to the early inflammatory stage of atherosclerosis.

Adult ; Arteries ; cytology ; drug effects ; metabolism ; Cells, Cultured ; Fluorescent Antibody Technique ; Humans ; In Vitro Techniques ; Inflammation ; chemically induced ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Tobacco Smoke Pollution ; adverse effects

Objective To evaluate the clinical efficacy of the local drug resistance spectrum antibiotics and foreign guideline in the treatment of patients with community acquired pneumonia(CAP). Methods A prospective,randomized,single blind,and positive drug parallel controlled design was used in the treatment. CAP patients with no underlying disease outpatients and inpatients<48 hours were selected as the research object. The patients in the trial group were given sensitive local drug resistance spectrum antibiotics: moxifloxacin,400 mg and 1 times a day. The patients in the control group were given azithromycin tablets(each 500 mg,once daily) promulgated by the 2007 version of the IDSA / ATS adult CAP guideline. Results There were 106 cases of CAP patients,of which 77 cases completed treatment,including 39 cases in the experimental group and 38 cases in the control group. There were significant differences in the clinical efficacy and bacterial clearance rate between the two groups,with the clinical efficacy of 89.7% and 68.4%(P < 0.01),the bacterial clearance rate of 87.9% and 54.5%(P < 0.05),respectively. Conclusion The clinical efficacy of drug resistant spectrum sensitive antibiotics in the treatment of CAP in Kunming was better than that of IDSA/ATS. Clinicians should pay attention to the characteristics and composition of resistance of common pathogenic bacteria in our country during the study and reference from foreign guideline,and adjust the therapeutic regimen according to the changes of the local drug resistance monitoring data rather than copy the recommended treatment plan by foreign countries.

OBJECTIVETo elevate the immunological effect of subunit influenza vaccine in infants and aged people (over 60) using liposomal adjuvant in the context of its relatively low immunity and to investigate the relation between vaccine antigens and liposomal characteristics.

METHODSSeveral formulations of liposomal subunit influenza vaccine were prepared. Their relevant characteristics were investigated to optimize the preparation method. Antisera obtained from immunizinged mice were used to evaluate the antibody titers of various samples by HI and ELISA.

RESULTSLiposomal trivalent influenza vaccine prepared by film evaporation in combinedation with freeze-drying significantly increased its immunological effect in SPF Balb/c mice. Liposomal vaccine stimulated the antibody titer of H3N2, H1N1, and B much stronger than conventional influenza vaccine. As a result, liposomal vaccine (mean size: 4.5-5.5 microm, entrapment efficiency: 30%-40%) significantly increased the immunological effect of subunit influenza vaccine.

CONCLUSIONThe immune effect of liposomal vaccine depends on different antigens, and enhanced immunity is not positively correlated with the mean size of liposome or its entrapped efficiency.

Animals ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza B virus ; immunology ; Influenza Vaccines ; administration & dosage ; immunology ; Liposomes ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections ; prevention & control ; Specific Pathogen-Free Organisms ; Vaccines, Subunit ; administration & dosage ; immunology

OBJECTIVETo study the distribution of genomovars and microevolution of Yersinia pestis in the Qinghai-Tibet Plateau.

METHODSPrimer pairs targeting the twenty-two different regions(DFRs) were designed for detecting the presence or deletion of each DFR in 297 strains isolated from the Qinghai-Tibet Plateau.

RESULTS9 genomovars, i. e. Genomovar 1, 5, 6, 7, 8, 10, 11, new type and Ype-ancestor were identified in the Marmota himalayana plague focus of the Qinghai-Tibet Plateau. Among these genomovars, genomovar 5,8 and 10 were dominant types. The total rate of the three genomovars was 80.6% (204/253) and the genomovars in different regions were different. All of 44 strains of Y. pestis in the Microtus fuscus plague focus of the Qinghai-Tibet Plateau belonged to genomovar 14.

CONCLUSIONThe distribution of genomovars of Y. pestis in the Qinghai-Tibet plateau had remarkable characteristics geographically. Based on the distribution of genomovars of Y. pestis, the routes of transmission and microevolution of Y. pestis were proposed.

Biological Evolution ; China ; Geography ; Humans ; Plague ; transmission ; Yersinia pestis ; genetics

OBJECTIVETo study the value of high-frequency ultrasound in the diagnosis of duchenne muscular dystrophy diseases (DMD) in children.

METHODSEight children with DMD were enrolled as DMD group and 10 healthy children as the control group. The echogenicity of the rectus femoris muscle and the gap between the gastrocnemius and soleus muscles in the two groups were detected by high-frequency ultrasound.

RESULTSCompared with the control group, rectus femoris and gastrocnemius muscles in the DMD group showed increased echogenicity and their muscle fibers were arranged irregularly, and the gap between the gastrocnemius and soleus muscles became wilder (P<0.01).

CONCLUSIONSHigh-frequency ultrasound is valuable in the diagnosis of DMD.

Child ; Child, Preschool ; Female ; Humans ; Male ; Muscular Dystrophy, Duchenne ; diagnostic imaging ; Ultrasonography

BACKGROUNDWe investigated whether and to what extent the ratio between circulating fibrinogen (Fg) and its degradation products (FDP) reflects the severity of coronary artery disease (CAD) in type 2 diabetic patients.

METHODSPlasma levels of Fg and FDP were determined, and Fg/FDP ratio was calculated in 344 consecutive patients with type 2 diabetes and chest pain on exertion undergoing coronary angiography. The severity of CAD was evaluated by the number of significant CAD (>50% luminal diameter narrowing) and Gensini score.

RESULTSPlasma Fg was higher, but Fg/FDP ratio was lower in patients with significant CAD (n = 255) compared with those without (n = 89), due to a disproportionate increase in FDP. Fg and FDP correlated positively, while Fg/FDP ratio negatively, with the number of diseased coronary arteries and the tertile of Gensini score (all P values for trend < 0.01). After adjusting for age, sex, risk factors for CAD, lipid profiles, glycosylated hemoglobin A1c, creatinine, leukocyte count, and high-sensitivity C-reactive protein, Fg/FDP ratio remained an independent determinant for multivessel coronary disease (MVD) (odds ratio [OR], 0.869; 95% confidence interval [CI], 0.788-0.958, P = 0.005) and high tertile of Gensini score (OR, 0.797, 95% CI, 0.682-0.930, P = 0.004). The area under the curve of Fg/FDP ratio was larger than that of Fg for predicting the presence of MVD (0.647 vs. 0.563, P = 0.048) and Gensini score ≥ 30 (0.656 vs. 0.538, P = 0.026).

CONCLUSIONSElevated plasma Fg and FDP level and reduced Fg/FDP ratio are associated with presence of CAD, and Fg/FDP ratio is superior to Fg in reflecting severe coronary atherosclerosis for patients with type 2 diabetes.

Aged ; Coronary Angiography ; Coronary Artery Disease ; blood ; diagnosis ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; metabolism ; Female ; Fibrin ; metabolism ; Fibrinogen ; metabolism ; Humans ; Male ; Middle Aged

In this work,a highly sensitive electrochemical biosensor for the detection of trace adenosine triphosphate (ATP) was proposed.The biosensor was based on porous anodic alumina (PAA) and SiO2 nanoparticles combining with several oligonucleotides to construct sandwich structure.It was characterized by scanning electron microscopy,fluorescence microscopy,differential pulse voltammetry and electrochemical impedance spectroscopy,which conformed to the reliability of the biosensor fabrication and the feasibility of the detection.In the presence of ATP,the sandwich structures could be destroyed.The variation of the current was directly corresponding to the amount of the ATP.The application of SiO2nanoparticles could effectively reduce the background and increase the sensitivity of the biosensor.The calibration curve of ATP was obtained in the range of 0.025-0.900 nmol/L with the detection limit of 13 pmol/L (S/N=3).Also,the biosensor exhibited a good specificity.Besides,the sensor was constructed easily and possessed excellent regeneration ability.The proposed biosensor was applied in detection of real sample such as mice blood.Therefore,the proposed ATP-sensing biosensor could be expected to be applied in clinical,pharmaceutical and environmental detection.

OBJECTIVE: To explore the pathogenesis of two fetuses from one family affected with Joubert syndrome (JS). METHODS: Whole exome sequencing was employed to screen potential mutations in both fetuses. Suspected mutations were verified by Sanger sequencing. Impact of intronic mutations on DNA transcription was validated by cDNA analysis. RESULTS: Two novel TCTN1 mutations, c.342-8A>G and c.1494+1G>A, were identified in exons 2 and 12, respectively.cDNA analysis confirmed the pathogenic nature of both mutations with interference of normal splicing resulting in production of truncated proteins. CONCLUSION The genetic etiology of the family affected with JS has been identified.Above findings have enriched the mutation spectrum of TCTN1gene and facilitated understanding of the genotype-phenotype correlation of JS.
Abnormalities, Multiple ; diagnosis ; genetics ; Cerebellum ; abnormalities ; Eye Abnormalities ; diagnosis ; genetics ; Humans ; Kidney Diseases, Cystic ; diagnosis ; genetics ; Membrane Proteins ; genetics ; Mutation ; Pedigree ; Retina ; abnormalities ; Whole Exome Sequencing