Objective To investigate the expression and clinical significance of proapoptotic genes HtraA2 in acute myeloid leukemia.Methods 78 cases of AML patients were divided into newly diagnosed AML group,complete remission group and hard flag group,and another 25 cases treated at the same period were set as the control group.The boue marrow and peripheral blood samples were collected from all the groups for total RNA extraction and detection of expressed HtrA2.The HtrA2 expressions were compared among thc groups.Finally 17 patients were followed up for 1～56 months.Results The HtrA2 expression levels of 3 groups were significantly different (x2 =35.13,P ＜ 0.05),with the ratio of maximum to minimum values up to 68.76.There were no statistically significant differences in the relative expression of gcnes HtrA2 among the FAB type (F =0.004,P ＞ 0.05).HtrA2 gene expression after treatment was significantly higher than before treatment in the patients followed up (P ＞ 0.05).HtrA2 gene might affect the survival time of patients (Wald =4.979,P ＜ 0.05),but age and gender had no influence on survival states (Wald =2.426 and 0.833,P ＞ 0.05).Survival curve analysis showed that the median smvival time was 34.50 months in the patients followed up.Conclusion The expression level of HtrA2 can be beneficial for the diagnosis,treatment and prognostic evaluation of AML.

Objective To construct a novel M.tb DNA vaccine (p846) co-expressing mycobacterial triple antigens including Rv3615c, Mtb10.4 and Rv2660c, and evaluate its cellular immune response and protective efficacy against tuberculosis infection in BALB/c mice. Methods We constructed the p846 by using the cloning technology. The 6- to -8-week old female BALB/c mice were randomly divided into 4 groups: p846, pcDNA3.1, PBS and the BCG group. All mice were administrated intramuscularly with 50 μg recombinant plasmids at 0, 2, 4, 6 week. A single dose of BCG was injected subcutaneously in the BCG group. Two weeks after the final immunization, 10 mice in each group were used for cell proliferation, ELISPOT and FCM assay, BCG challenge experiment and HE staining of lung were performed at 4, 6 weeks later, respectively. Results The p846 vaccine could effectively induce the specific T cell proliferation(P < 0.001) and increase the numbers of IFN-γ+T cells(P <0.001), compared with those in the PBS group and the vector conreol group. The mouse lung tissue presented very mild lung inflammation in the p846 group, compared with other groups. Conclusion Vaccine p846 could not only induce strong cellular immune response, but also efficiently protect BALB/c mice against M.tb infection.

Objective To analyze and determine the clinical, molecular pathology and genetic features of a Chinese family with dystrophinopathy. Methods Clinical data of the proband and his family members were collected. Immunohistochemistry staining was performed on muscular biopsy tissues with antimerosin, emerin and the N, C and central rod domains of dystrophin. Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes. Multiplex ligation-dependent probe amplification (MLPA) was used to test Duchenne muscular dystrophy (DMD) gene to determine the ways and sites of genetic mutation, and analyze the relationships between genotype and phenotype. Results Patients from this family were clinically diagnosed as muscular dystrophy, and they presented serious manifestations although the immunohistochemistry analysis for the proband exhibited partial loss of dystrophin staining, and positive expression with merosin and emerin. Further test with MLPA detected the loss of exons 45--54 in DMD gene in the proband, while his mother had heterozygositic loss in exons 45--54. Conclusions The losses of exons 45--54 in the proband are all derived from his mother, who carries genetic mutation with normal phenotype. He has been diagnosed as dystrophinopathy. At the same time, his partial loss of dystrophin is not parallel to the out-of-frame mutation of the gene and his severe clinical manifestations. Abnormal expression of dystrophin is the pathological basis for dystrophinopathy phenotype. Its clinical outcome depends not only on the degree of the protein expression, but also on the function of the sites where the DMD gene less occurs.

Objective To clone 2-hydroxyacid dehydrogenase (HADH) gene from Ketogulonigenium vulgare(KGV) Y25 and investigate its expression , purification, and enzymatic characterics .Methods The hadh gene was amplified from ketogulonigenium vulgare Y 25 and cloned into the expression plasmid pITG .The recombinant plasmid was transformed into Escherichia coli BL21(DE3).HADH was then successfully expressed with induction .To explore its enzymatic characteris-tics,HADH was purified by Ni +exchange chromatography .Results HADH constituted more than 50% of the total cell proteins analyzed by SDS-PAGE,with a relative molecular mass of about 35 ×103.With 2-keto-gulonic acid(2-KGA) as substrate, the optimal pH of HADH was at 8.0 ,while the optimal temperature of the purified HADH was at 45℃.Mean-while, such metal ions and chelating agents as Cu 2+, Ca2+, Mg2+, EDTA, and DEPC exerted little effect on enzymatic activities.The maximum initial activity of the enzyme towards 2-KGA reached 27 U/mg, and the Km was calculated as 2.6 mmol/L.The results of in vivo enzyme activity assay showed that HADH could metabolize 2-KGA.Conclusion The HADH gene form Y25 is successfully expressed in E.coli BL21 ( DE3 ) and the enzymatic characteristics of HADH are explored, which will facilitate subsequent studies on sorbose metabolic pathways and sugar acid conversion .

Infantile neuroaxonal dystrophy (INAD) is a rare autosome-recessive disease characterized by progressive motor and cognitive regression.The PLA2G6 gene is its causative gene,which encodes calcium-independent phospholipase A2 enzyme (iPLA2-VIA).The diagnosis of INAD is difficult because of its clinical heterogeneity,and the rate of misdiagnosis is high.The purpose of this study is to describe the clinical characteristics,molecular genetics,treatment and prognosis of INAD to improve the acknowledgement of INAD in medical workers and to help make an early diagnosis of INAD.

Super hydrophobic interface modified with silver nanoparticles was fabricated for the detection of pesticide residues.By using a chemical reduction method,silver nanoparticles were deposited on the substrate surfaces with different microscopic pore structures.Two kinds of composite substrates,including regular stainless steel mesh and cellulose polyester film,were used.The pre-treatment of the substrate with fluoridated reagents was used to form a super hydrophobic interface,which made the target molecules on the surface concentrate effectively.The surface with the cellulose polyester substrate was used to detect Rhodamine 6G (R 6G) effectively with surface enhanced Raman scattering (SERS) technique.The results showed that the detection hmit was 10-16 mol/L.In addition,the surfaces based on the stainless steel mesh and cellulose polyester substrate were used to detect trichlorfon pesticide with detection limits of 1 × 10-15 mol/L and 1 × 10-16 mol/L,respectively.

A reliable method for simultaneous determinition of eleven representative components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin) in combination of chromatographic fingerpint analysis for Reduning injection was developed by ultra high-performance liquid chromatography (UPLC). The method was performed on an Agilent ZORBAX SB-C18 anlytical column (3. 0 mm x 100 mm, 1. 8 µm) with a guard column of Agilent UPLC Guard ZORBAX SB-C18 (3.0 mm x 5 mm) at the column temperature of 30 °C. The gradient mobile phase consisted of acetonitrile (A)-0. 1% phosphoric acid (B) with a flow rate of 0. 4 mL . min-1. The injection volumn was 2 µL. The detection wavelengths were set at 324 nm and 238 nm for quantit tive analysis and 225 nm for fingerpint chromatography. Neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin were baseline seperated with good linearity relationships (r >0. 999) between concentration and peak areas over the linear ranges. The average recoverys of the investigated compounds were 103.5%, 100. 2%, 103. 3%, 102. 8%, 101. 3%, 102. 8%, 97. 36%, 99. 62%, 98. 16%, 102. 8%, 99. 27%, respectively. Reduning injection of forty-five batches was analyzed by UPLC finge print chromatography. Thirty batches were selected to generate the reference fringerprint chromatography with fourteen common peaks. The similarity values between the reference fringerprint chromatography and the remaining fifteen batches were higher than 0. 99. The developed method was fast, accurate and sensitive. It could be used as a reference for the quality control of multiple components determination and fingerprint chromatography for Reduning injection in future.
Chlorogenic Acid ; chemistry ; isolation & purification ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Glucosides ; chemistry ; isolation & purification ; standards ; Iridoids ; chemistry ; isolation & purification ; standards ; Quality Control ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity ; Time Factors

Objective To establish a risk warning model for surgical site infection(SSI), provide support for screening high risk population and finding suspected cases.Methods Clinical data of 5 067 patients who underwent abdominal surgery in 6 domestic hospitals from January 2013 to December 2015 were collected retrospectively, all cases were randomly divided into modeling group and validation group according to a 6:4 ratio, warning model was established by employing logistic regression, the area under the receiver operating characteristic curve (AUC) was used to evaluate discriminant ability of evaluation model, the maximum Youden index was as the optimum cut-off point.Results For the warning model of high-risk patients, AUC was 0.823, sensitivity and specificity were 78.81% and 74.33% respectively, positive predictive value and negative predictive value were 19.67% and 97.78% respectively.For the discriminant model of suspected infection cases, AUC was 0.978, sensitivity and specificity were 93.38% and 95.62% respectively, positive predictive value and negative predictive value were 62.95% and 99.45% respectively.Conclusion The early-warning model established in this study has better discrimination ability, which can provide a reference for the development of early warning and discrimination of healthcare-associated infection information system.

Objective To evaluate five years results of laser in situ keratomileusis(LASIK)for correction of myopia.Design Ret- rospective case series.Participant 64 patients(126 eyes)with myopia received operation of LASIK.Method 126 myopic eyes were treated with LASIK.Based on the preoperative spherical equivalent refraction(SER),the patients were divided into Group A,B and C(-3.00D～6.00D,-6.25D～-10.00D and -10.38D～-19.88D).Before and after operation,the visual acuity,refraction,anterior seg- ment,funduscopy,intraocular pressures of patients were measured,and the patients were followed-up five years.Main Outcome Measure Visual acuity,intraocular pressure,refraction and complication.Result At five years,in Group A,B,C,uncorrected visual acuity was≥1.0 in 87.2%,69.1%,31.3% patients,and 100%,98.2%,75% had vision of≥0.5.After five years of operation,the re- fraction was -0.70D?0.52D,-1.06?0.13D,-2.46?2.14D in three groups respectively,there was no significant difference between Group A and B(P=0.23);But there was significant statistical difference between Gruop A and C(P=0.008),Group B and C(P=0.021)respec- tively.At the fifth-year,68.3%,45.0% and 23.5% of refraction in Group A,B,C were within?1.00D.Intraocular pressure at the fifth year was 11.91?2.35mmHg,11.31?2.20 mmHg and 9.24?2.20 mmHg respectively.In Gruop B and C,one patient complained glare re- spectively.In Gruop B,one patient's intraecular pressure was elevated after using glueocorticoid.In Group C,one eye was complicated with rbegmatogenous retinal detachment at the second year after LASIK.Conclusion In the follow-up of five years,LASIK is effective and safe.The results in middle and high myopia are better than that in extreme high myopia.(Ophthalmol CHN,2006,15:312-314)

OBJECTIVETo investigate the effect of sensory neuropeptide substance P (SP) on the differentiation of cultured epidermal stem cells (ESC) in vitro,with in vitro cultured ESC as the platform.

METHODSESC from newborn Wistar rats were isolated, purified by repeated passages in culture. SP was added for stimulation when ESC clone grew. Immunohistochemistry staining with K14 antibody, and flow cytometry (FCM) was performed at 0, 24th, 48th, 72nd, 96th, 144th, 192nd, 240th, 288th, 336th, 384th, 432nd post differentiation hours (PDH) to identify the cell groups and to detect if there were transient amplifying cells (TAC) among the cells.

RESULTSESC in culture formed large colonies after SP treatment with positive staining for K14, indicating that they were TACs. The results of FCM indicated that when ESC were stimulated by SP, TAC colony formation occurred and the cell number increased in a constant speed.

CONCLUSIONESC could differentiate into TAC by neuropeptide SP induction, and the number of ESC kept on a certain level during the process.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Flow Cytometry ; Male ; Rats ; Rats, Wistar ; Sensory Receptor Cells ; chemistry ; Stem Cells ; cytology ; Substance P ; pharmacology