To decrease the oxygen content in the cell is a key method to improve hydrogen production in Chlamydomonas reinhardtii.A new approach was developed by transforming the leghemoglobin gene lba,which has high affinity to oxygen,into the chloroplast of C.reinhardtii to get a low dissolved oxygen in the cell and result into improvement of H2 ase activity and H2 yield. The results showed that lba was successfully transformed into the chloroplast of C.reinahrdtii strain 849 and did not affect its growth significantly. The work paved the road for further regulation of lba expression in the chloroplast to improve of hydrogen production of C.reinahrdtii.

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.

Objective To explore the relationship between the diabetic retinopathy (DR) and the carotid artery intima-media thickness(IMT) in type 2 diabetic patients to reveal the relationship between macroangiopathy and microangiopathy in diabetic patients further. Methods One hundred and tweenty-three diabetic cases in patient chosen from 2008 to 2009 were divided into diabetic retinopathy group(DR) and non-diabetic retinopathy group(NDR) by fundus examination. The patients were asked about their disease history including durations, smoking and so on. Meanwhile the carotid artery IMT, systolic pressure (SBP), diastolic pressure (DBP), serum total cholesterol, triglycerides, high density lipoprotein (HDL)-cholesterol, low density lipoprotein (LDL)-cholesterol, glycosylated hemoglobin(HbAlc), body mass index(BMI) were measured of all the cases. The incidence of increased carotid artery IMT was cmpared with χ2 test, as well as the average IMT between the two groups, the influencing factors artery IMT was 50.98%(26/51) in DR group, and 33.33%(24/72) in NDR group, having a statistically significant showed the diabetic retinopathy risk factors were smoking(χ2=6.20, P<0.05), duration(t=-4.13, P<0.01). carotid artery IMT(t=-2.21, P<0.05), SBP(t=-2.37, P<0.05), and HDL-cholesterol(t=4.49, all P<0.01). 12.77, all P<0.01), carotid artery 1MT and smoking(χ2=6.05,4.15, all P<0.05). Conclusions Type 2 diabetic patients complicated with DR have a prominent increase in IMT thickening proportion and average IMT, which reveals the relationship between the DR and the IMT.

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

OBJECTIVETo study a feasible method of measuring right ventricular pressure by catheterization in mice.

METHODSMeasuring the right ventricular pressure and the pulmonary artery pressure by homemade PE pipe through venous cannula in external jugular vein, using catheterization in mice with powerlab multimodal biometric signal recording system.

RESULTSForty-six out of 51 mice were experimented with this method smoothly and got a total success rate of 90.2%. Thirty of 33 normal mice and 16 of 18 mice with pulmonary arterial hypertension (PAH) were catheterized successfully. The right ventricular pressure were as follow: systolic blood pressure: (23.4 +/- 5.7) mmHg in normal group vs (32.2 +/- 2.8) mmHg in mice with PAH, diastolic blood pressure: (3.7 +/- 2.6) mmHg vs (3.8 +/- 2.0) mmHg, mean pressure: (12.0 +/- 3.7) mmHg vs (14.9 +/- 2.3) mmHg. After autopsy for those 5 failed cases, we found that 2 cases were into the inferior vena cava, another 2 cases pierced the right auricle and the last one punctured the axillary vein into the chest wall.

CONCLUSIONMeasuring the right ventricular pressure through venous cannula in external jugular vein with homemade PE pipe in mice gets not only a high success rate but also help to save time. Moreover, this method can be popularized easily. It is a good and feasible method for measuring right ventricular pressure in mice.

Animals ; Cardiac Catheterization ; methods ; Jugular Veins ; Male ; Mice ; Mice, Inbred BALB C ; Ventricular Pressure

Objective To evaluate the sensitivity and specificity of different HbA1C cutoff points for diabetes diagnosis in high risk outpatients in Harbin.Methods A total of 2 122 high risk outpatients(male 1 032 and female 1 090)for diabetes screening in the Fourth affiliated Hospital of Harbin Medical University from April 2013 to February 2015 were included in this study, with the average age of(49.26±13.00)year. Oral glucose tolerance tests(OGTT)were conducted and HbA1C levels were examined in these patients. The sensitivity and specificity of different HbA1C cutoff points were calculated and a receiver operator characteristic(ROC)curve was then built.Results The average level of HbA1C in these subjects was(6.45±1.72)%. The prevalence of diabetes was 41.85%. The area under ROC curve(AUC)was 0.89 with the optimal cutoff point of HbA1C 6.0% and 0.68 for the highest Yonden index. The sensitivity and specificity of HbA1C 6.0% were 84.01% and 83.67% respectively. The sensitivity and specificity of HbA1C 6.5% were 62.84% and 95.92%, respectively. The AUC of HbA1C≥6.5% was 0.732. Conclusion HbA1C works well as the diagnostic standard for diabetes in high risk outpatients of Harbin city. The cutoff point of HbA1C 6.0% is suitable for screening diabetes in high risk population, and HbA1C 6.5% is appropriate for diabetes diagnosis, with high sensitivity and specificity.

The prokaryotic expression plasmid pET-30a(+)/sBLyS was constructed and transformed into E. coli BL21 (lambda DE3). The recombinant protein was found to be highly expressed by the plight of soluble part and inclusion body. For the sake of enhancing the proportion of the soluble part, inducement at 16 degrees C for 12 h was ascertained. The expressing product was then purified by Ni2+ affinity chromatography gel. PI of the recombinant human sBLyS(rhsBLyS) is about 7.1-7.3 and it assembles into a homotrimer. The effect of rhsBLyS on B lymphocytes by MTT method told us the B lymphocytes' proliferating capacity dose depended on concentration and also stimulating time of the rhsBLys. With rhsBLyS(2 micrograms/mL) stimulating 3 days, B lymphocytes can proliferate the most.
B-Cell Activating Factor ; B-Lymphocytes ; drug effects ; Escherichia coli ; genetics ; Humans ; Isoelectric Point ; Lymphocyte Activation ; drug effects ; Membrane Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Recombinant Proteins ; biosynthesis ; Temperature ; Tumor Necrosis Factor-alpha ; biosynthesis ; isolation & purification ; pharmacology

Objective Since it is a common problem for medical practice that paper instructions for medicine information are difficult to update and inconvenient to search ,this article is to provide convenient service of pharmacy information for clini-cal practice by developing medicine information search software in our hospital .Methods According to the practical needs of clinical study and pharmacy research ,we took appropriate measures to collect data ,categorize contents ,scan pictures ,polish images and extract information for the medicine instructions .Based on the above work ,we applied Oracle 10G to establish medicine information database for our hospital and utilized Java to design medicine information search software .Results Our search software includes 1 201 kinds of medicines ,taking up 98 .85% of the medicines in our catalogue .The users can use four different search methods (including common name search ,commodity name search ,fuzzy search and key word search) to gain the written material and photographic information of medicine instructions .Conclusion With the help of the software of Oracle and Java ,the newly designed software for medicine information search can provide convenient pharmacy information service for clinical practice .

OBJECTIVETo explore the distribution characteristics of the types of M protein gene (emm) in group A streptococcus (GAS) isolated from children in Beijing in the year 2011.

METHODSDuring May to July in 2011, a total of 3315 patients who were diagnosed scarlet fever or pharyngeal infection by doctors in pediatric outpatient and emergency units of 36 hospitals, were selected as subjects. Their throat swab samples were collected and isolated the strains of GAS. Gene emm was then amplified and sequenced by PCR method, and the differences in types of gene emm between different populations and diseases were compared.

RESULTSA total of 633 strains of GAS were isolated from the 3315 throat swab samples, 610 strains out of which were gene emm positive and were recruited in the study. Out of the 610 recruited strains, 448 (73.4%) were isolated from scarlet fever patients, the other 162 (26.6%) were isolated from pharyngeal infection patients; 397 (65.1%) were from urban, the other 213 (34.9%) were from suburb; 240 (39.4%) were from patients aging between 1 - 5 years old, the other 369(60.6%) were from patients aging 6 - 18 years old. A total of 8 types of gene emm (scarlet fever: 6 types, pharyngeal infection: 4 types) and 21 subtypes of gene emn (scarlet fever: 16 subtypes, pharyngeal infection: 10 subtypes) were identified. Three new subtypes were found in the study, naming emm1.63, emm12.62 and st5144.20. Among them, emm1.63 was found both in scarlet fever and pharyngeal infection patients, while emm12.62 and st5144. 20 were only found in pharyngeal infection patients. Among all the types of gene-emm, emm12 accounted for the highest percentage as 80.5% (491/610) and then followed by emm1 (18.0% (110/610)). Among all the subtypes, the dominant subtype was emm12.00, accounting for 69.0% (421/610), following by emm1.00 (16.9% (103/610)) and emm12.19 (6.1% (37/610)). All the above types and subtypes of gene emm were the most prevalent strains in scarlet fever patients and pharyngeal infection patients. Significant differences in the distribution of prevalent strains were observed among various aging patients and regions. The constituent ratios of emm1, emm1.00 and emm12.19 were higher in patients from suburb (emm1: 22.1% (47/213), emm1.00: 19.2% (40/213), emm12.19: 8.0% (17/213)) than those in urban areas (emm1: 15.9% (63/397), emm1.00: 15.6% (62/397), emm12.19: 5.0% (20/397)). The difference showed statistical significance (P < 0.05). The constituent ratio of emm1.00 was higher among patients aging 6-18 years old (19.2% (71/369)) than those aging 1 - 5 years old (13.3% (32/240)). The difference also showed statistical significance (χ(2) = 8.45, P < 0.05).

CONCLUSIONAmong the types of gene emm in GAS isolated from children in Beijing in year 2011, the most prevalent two were emm12 and emm1, and the most prevalent emm subtypes were emm12.00, emm1.00 and emm12.19. A significant difference in their distribution between various aging patients and isolated places can be obviously found.

Adolescent ; Antigens, Bacterial ; classification ; genetics ; Bacterial Outer Membrane Proteins ; classification ; genetics ; Carrier Proteins ; classification ; genetics ; Child ; Child, Preschool ; China ; Female ; Genes, Bacterial ; Genotype ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Streptococcus pyogenes ; genetics ; isolation & purification

Objective To evaluate the feasibility of quantitative analysis software for myocardial contrast echocardiography (MCE) in assessment of myocardial perfusion.Methods According to coronary occlusion and reperfusion at different times,rabbits were divided into two groups:15 min occlusion / 30 min reperfusion (group Ⅰ) and 120 min occlusion / 60 min reperfusion (group Ⅱ).MCE was performed on all rabbits at baseline,occlusion and after reperfusion,and its images were analyzed by a new quantitative analysis software based on eliminating particle swarm optimization (EPSO) clustering algorithm,by which obtain myocardial perfusion parameters.Results (1) The values of calibrated contrast intensity (CI) in risk segments of Groups Ⅰ and Ⅱ were significantly lower than those at baseline during occlusion (t =5.104 and t =4.327,P＜0.01).After reperfusion,calibrated CI in risk segments significantly improved in Group Ⅰ (t =2.933,P＜0.01) while those remained unchanged in Group Ⅱ (P＞0.05).(2) The areas of red-coded region in color-coded map and myocardial infarction in triphenyl-tetrazolium chloride (TTC) were (21.4±12.3)% and (18.0±9.5)%,respectively.The correlation between color-coded image and TTC was 0.89 (P＜0.01).(3) The histogram in all risk segments was skew distribution during occlusion.After reperfusion,the histogram in Group Ⅰ was normal distribution while that was still skewed distribution in Group Ⅱ.Conclusion The MCE image analysis software based on EPSO clustering algorithm in the quantitative assessment of myocardial microperfusion and identification of myocardial perfusion abnormalities was feasible and of high value.