Though prokaryotic cells could hardly express recombinant human beta nerve growth factor (rhNGF-?) with a proper three-dimensional conformation, using of E. coli as a host for industrial production of rhNGF-? is controversial. Recombinant human beta NGF was expressed in E. coli and was refolded in vitro. The isolated products was shown to be consistent with those expressed and secreted by CHO cells in biochemical characters by SDS-PAGE, RP-HPLC, mass spectrometry, N terminal analysis and bioassay determined using DRG and PC12 cells. The products can be acquired with 95% purity, 1.8 ng/U biological activity from both expression system,which remain invariable biological activity when lyophilized in excipient and store at 37℃?2℃, RH 75%?5% for 3 months. Moreover, the product refolded from inclusion bodies of E. coli shows the predominance in homogeneity and the lower cost.

Background Diabetiretinopathy(DR) icommon ophthalmivasculadisease which can lead to blindness,and itpathogenesiicomplex.C-peptide iby-producin insulin synthesis,and loof studieindicate thaC-peptide iclosely related with diabetimicrovasculacomplications.Objective Thistudy wato explore the clinical significance of the detection of C-peptide to assesthe prognosiof non-proliferative DR(NPDR).Methodretrospective case-observational study wadesigned.The clinical datof 205 casewith NPDwere collected in Wuhan Puren Hospital from January 1,2009 to January 1,2010.The patientwere diagnosed by fundufluorescein angiography (FFA) and the stageof theidiseasewere determined based on the Dstaging developed by the Chinese Ophthalmological Society fundustudy group.Twenty-seven-month follow-up waperformed fothe patientto record the parameterof theiblood biochemistry,changein theiphysical conditions,and changein theiblood pressure and DM.The patientwere assigned to the PDgroup and NPDgroup athe end of the follow-up.The independenrisk factofoNPDwaanalyzed using the univariate analysis,and multivariate logistiregression analysiwaused to evaluate the correlation of varioufactorwith pooprognosiof NPDR.Receiveoperating characteristi(ROC) curve wadrawn to assesthe prediction efficiency of C-peptide fothe prognosiof NPDR.ResultThe clinical datof 205 patientwith NPDwere reviewed,and 15 patientfailed to show up fovisitarate of 7.3％.One hundred and ninety patientwith NPDfinished follow-up with mean age of (41.54± 8.15)years.Of the 190 patients,69 patient(36.3％) advanced to PDR,and 121 patient(63.7％) were NPDduring the follow-up.Significandifferencewere seen in age,hypertension history,smoking history,DM course and C-peptide levelbetween the PDgroup and PDgroup (age:=13.78,P＜0.05 ; hypertension history:x2 =21.57,P＜0.05 ;smocking history:x2 =26.96,P＜0.05;DM course:t=6.89,P＜0.05;C-peptide:t=20.50,P＜0.05).pooprognosiwacorrelated with age (OR:1.04,95％ CI:1.01-1.07),hypertension history (OR:1.05,95％ CI:1.02-1.08),smokinghistory (OR:1.07,95％CI:1.04-1.11),DM course (0R:1.18,95％CI:1.13-1.25) oC-peptide conten(0R:1.09,95％ CI:1.06-1.12).The areundethe ROcurve wa0.835 (P＜0.05,95％ CI:0.818-0.843) when the C-peptide waincluded,buwalarge than 0.769 (P ＜ 0.05,95％ CI:0.754-0.780) when Cpeptide waexcluded.ConclusionThe levelof blood C-peptide are lowein the PDpatientthan those in the NPDpatients,and the lowelevel of blood C-peptide in the NPDpopulation increasetheirisk of developing PDR.Testing foblood C-peptide level can benefithe prognosiof NPDR.

This study was aimed to investigate active ingredients from A mp e lop s is me galop hylla of protein expression by using two-dimensional gel electrophoresis (2-DE). Methods: isn this study, proteinsfrom the tender leaves of A mpelopsis megalophylla were precipitated by Tris-phenol extraction and ammonium acetate methanol precipitation. Then using 2-DE technology to isolate protein.Finally, 2-DE was analyzed by the scanner, and got protein characteristic maps. Result:According to the differences in ratio of gray value, the seven differential protein points were chosen to identify the mass spectrum. At present, two proteins were identified, namely the hypothetical protein and the unnamed protein product. Conclusion: the application of 2-DE technology for A mpelopsis megalophyllathe active ingredient of protein expression at the level of molecular research laid a good foundation.

Objective To analyze the CT findings of lung cystic echinococcosis with infections, and to assess the value of CT examination. Methods The CT characters of 23 cases of pathologically proved lung cystic echinococcosis were analyzed and CT findings were compared with pathology. Results All 23 cases of lung cystic echinococcosis appeared as single cystic lesions. Fifteen cases appeared in the right lung, and the other 8 in the left. The lesions located in lower lobe were more than those in the upper lobe. According to the CT examination, it could be divided into two types: (1) simple lesion (7 cases) appeared as irregular solid mass with different size, crude edge, and lobulation, and with fiber shadows in the lung field around the lesions. In 6 cases, curvilinear or spotty calcification occurred in the lesions. (2) echinococcosis as a bag (16 cases) appeared as a mass with cystic changes in lung tissue with different size and off-center cystic cavity. The lesions took the form of ball or irregular shape. The edge was crude. Around the focus, prickle-like or patchy changes were detected. Ruptured lesions showed “water snake” or “flow ribbon” signs in 4 cases. In 7 cases, calcification occurred on the cystic wall or extended into the inside contents. Conclusion CT could accurately demonstrate the location, appearance, internal structure, and adjacent situation of the cystic echinococcosis, providing valuable information for correct diagnosis and treatment.

Objective: To investigate the effects of estrogen and progesterone on the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR) in mouse uterus. Methods: 3-week-old immature female mice were randomly divided into 7 groups and treated with corn oil, estradiol (E2) of 1.5, 3.0, 10, 25 ng, progesterone (P) of 100 ?g and (E_2 10 ng + P 100 ?g)/mouse, respectively. After the treatment for 48 h, mouse uterus was collected to isolate total RNA. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA isoforms of VEGF and its receptors in mouse uterus. Results: Compared with control, both E_2 and P significantly increased the expression of VEGF164 and VEGF120 mRNA in mouse uterus. The expression of VEGFR2 mRNA, not VEGF1 mRNA, was decreased by E_2 treatment in a dose-independent manner. Conclusion: Both estradiol and progesterone up-regulated the expression of VEGF, but estradiol down-regulated the expression of VEGFR2 in mouse uterus.

This article introduced the development and application effect appraisal of Microbial Engineering CAI courseware for bio-engineering specialization. The courseware focuses on knowledge system integrity, content-rich and gives prominence to the key points. Pictures, animation and video, and audio effects are also utilized appropriately to achieving stimulate students interest in learning and then improve teaching and learning performance. The courseware concentrates on core content of the course, such as fermentation parameters detection and automatic control, and fermentation equipments. The courseware was manufactured using the Powerpoint software. Animation was established with Flash 4 software and the scanning pattern was edited using Adobe photoshop. And chapters of the courseware were composed and administrated using Courseware Master Software. A two-year survey showed that 85% of students satisfied with this courseware.

Aged ; Castleman Disease ; complications ; Hepatitis B ; complications ; Humans ; Liver Neoplasms ; complications ; virology ; Lymphoma, Non-Hodgkin ; complications ; Male

Objective To explore the feasibility and clinical significance of combined detection of human plasma and feces expression level of microRNA-92a-1 (miRNA-92a-1) as colorectal tumour screening marker.Methods From August to October in 2011,the feces and plasma samples of 60 colorectal cancer (CRC) patients,23 colorectal adenoma (CRA) patients and 30 healthy controls were collected.The expression level of miRNA-92a-1 was determined by quantitative reverse transcription-polymerase chain reaction.Mann-Whitney's U test was applied for the difference test between groups.Then,according to the receiver operating characteristic (ROC) curve analysis the cut-off point was determined and the sensitivity and specificity were analyzed.Results Compared with healthy controls,the expressions of miRNA-92a-1 in plasma of CRC and CRA patients increased (U=288.5 and 151.0,both P＜0.01).Compared with healthy controls,the expression of miRNA-92a-1 in feces of CRC patients increased (U=627.5,P=0.0199).According to ROC curve analysis,when the cut-off point was ＞1.22,the sensitivities of plasma miRNA-92a-1 in CRC patients and CRA patients were 85.0％ (51/60) and 73.9％ (17/23) respectively.The specificity of healthy controls was 76.7％ (23/30).When the cut-off point was ＞1.14,the sensitivities of feces miRNA-92a-1 in CRC patients and CRA patients were 31.7％(19/60) and 26.1％(6/23) respectively.The specificity of healthy controls was 90.0 ％ (27/30).Combined the detection results of plasma and feces,the sensitivities of miRNA-92a-1 in CRC patients and CRA patients was 88.3％ (53/60) and 82.6％ (19/23) respectively.The specificity in healthy controls was 73.3％ (22/30).The sensitivity was higher than that of single sample testing.Conclusions The sensitivity and specificity are high in the combined detection of plasma and feces miRNA-92a-1 expression level in CRC patients and CRA patients,miRNA-92a-1 may be a potential CRC screening marker.

Objective To evaluate the performance of DNA microarray for rapid detection resistance to rifampin and isoniazid in Mycobacterium tuberculosis clinical isolates and identify suitable target sites for molecular genetic test.Methods Twenty-four clinical Mycobacterium tuberculosis isolates were collected retrospectively from Shenzhen Center for Chronic Disease Control in 2009 and 127 isolates from project on anti-tuberculosis drug resistance surveillance in Shenzhen during 2007 to 2009.Drug susceptibility to rifampin and isoniazid of the stains were determined by DNA microarray,and results were compared to that obtained with reference proportion method drug susceptibility testing for sensitivity,specificity and accuracy.The consistency of microarray and phenotypic susceptibility testing was evaluated by Kappa test.Genetic mutations in rpoB,katG,inhA,regulatory region of inhA,and regulatory region of ahpC were investigated by DNA sequencing to assess proper loci for rapid molecular diagnosis.Results Compared against results of proportion method,the sensitivity,specificity and accuracy of the DNA microarray assay for rifampin resistance were 94.4％,97.5％ and 96.0％ respectively,and for isoniazid resistance were 79.1％,100％ and 86.8％ respectively.Mutations in resistance-determining region of rpoB were observed in 97.2％ (70/72) of the isolates resistant to rifampin,which contributed in the 531,526,516,511 and 533codon region.Mutations in katG315 codon,inhA-15,and ahpC regulatory region were found in 70.3％ (64/91),11.0％ (10/91) and 9.9％ (9/90) of the isolates resistant to isoniazid,respectively.Mutations of ahpC promoter region consists of ahpC-9 (4 strains),ahpC-10 (2 strains),ahpC-6 (2 strains),ahpC-12 (1 strain),and ahpC-32 (1 strain).Conclusions DNA microarray provided a rapid method for the detection of drug-resistant Mycobacterium tuberculosis isolates,and demonstrated good performance except less sensitive for isoniazid resistance.The mutations in ahpC regulatory region might be good target loci for detection of isoniazid-resistant Mycobacterium tuberculosis,so screening the region may significantly improve the sensitivity for molecular genetic tests.

The effects of several factors on the chlamydospores production of Trichoderma aureoviride T-33 during the fermentation were researched.Based on the results above, the orthogonal test was made to screen out the best prescription.The results showed that, the best single-factor conditions for the chlamydospores production of T.aureoviride T-33 were, liquid culture of oat powder, 30℃, pH4.0, 120r/min, 24 hours oscillate incubating as well as liquid culture volume of 80mL/bottle when the 250mL size triangle bottle was used.The result of orthogonal test showed that, the best prescription for temperature, pH and oscillating speed was 30℃, pH4.0 and 140r/min.3.37?10~ 7 spore/mL chlamydospores were obtained at this combined condition.