Search

Antibiotic residues in milk powder are public focusing concerns relevant to food safety. Long term exposure to antibiotic residues will lead to drug resistant strains in human body, reduce body's immunity, and bring health hazards to consumers. Sulfonamides, (fluoro)quinolones and macrolides are high frequency detecting antibiotic residues in matrix of milk powder. Rapid screening analysis of these residues and simultaneous confirmation with high confidence are valuable in practical scenarios of investigation testing, special monitoring program and so on. The direct analysis in real time (DART) ion source is an emerging technology of ambient pressure desorption ionization, it can directly complete the analysis of sample ionization and injection at the ambient pressure and open experimental environment, without changing the properties of the analyte itself. It became possible to analyze samples directly with minimal sample preparation steps or even without any sample pretreatment procedure with DART ion source. A hybrid high resolution mass spectrotmety, quadrupole orbitrap high resolution mass spectrometry (Q-Orbitrap HRMS) is a powerful tool in veterinary drug residue analysis with superior specificity, accuracy and sensitivity. In present study, DART ion source was hyphenated to Q-Orbitrap HRMS to perform rapid screening analysis and simultaneously unambiguous identification of the above mentioned antibiotic residues in milk powder. Sample was diluted in water and fast purified by filtering solid phase extraction column and then the filtrate was subjected to DART QOrbitrap HRMS analysis. Several key parameters of the DART ion source, such as ionization type and temperature, as well as injection speed of DART ion source were optimized to enhance the performance of the method. Then, a QuickStrip (QS) Module was selected with the helium as carrier gas and ionization temperature as well as injection speed were set to 400 ℃ and 1 mm/s, respectively. The DART ion source with optimized settings coupled to Q-Orbitrap HRMS was finally applied to screening analysis. Results showed that 16 commonencountered antibiotic residues including sulfonamides, (fluoro)quinolones and macrolides can be effectively detected with the screening concentration of 100 μg/kg. Recoveries of the analytes ranged from 72.5% to 106.7% and the false negative rates were all less than 5%. Real life sample analysis was finally conducted by the proposed method with an expected method performance, evidencing the potency of the developed method in risk monitoring in milk powder for veterinary drug screening analysis. It should be mentioned here that the screening concentration of 100 μg/kg is a compromised concentration for the sake of some less sensitive analytes to provide reliable determination of the antibiotic residues with the proposed procedures. It is, however, quite meaningful concentration to indicate the existence of high risk level of corresponding antibiotic residues which need to take actions. In total, the proposed method can play a role in risk monitoring laboratories and relevant industry institutions.

In this study, a large area of well preserved Zostera japonica dominated meadow was found in a coastal lagoon, Swan Lake, in Rongcheng of Shandong Province. Due to its unique geographical position and high biomass, this meadow may act as a typical Z. japonica bed in the coastal area of Shandong. From September, 2011 to October, 2012, an annual investigation was conducted on the Z. japonica and its habitats in east coast of Swan Lake, and the distribution of the Z. japonica and its habitats ecological characteristics were preliminarily understood. The major ingredients of sediments particles in the Z. japonica bed was sand (81%) and silt (14%). The C and N contents in the sediments were the highest in winter, and the C/N ratio was the highest in autumn. The shoot density, shoot height, and biomass of the Z. japonica were all significantly correlated with water temperature (P < 0.05). There was an obvious change in the Z. japonica growth among seasons, with the peak biomass obtained in August-September. The C and N contents and C/N in Z. japonica leaves also varied with seasons. The leaf C content was significantly higher in autumn than in spring and summer (P < 0.05), the leaf N content was significantly lower in summer than in spring and autumn (P < 0.01), whereas the leaf C/N ratio was significantly higher in summer than in spring (P < 0.05). The annual carbon sequestration by the Z. japonica in the Swan Lake was estimated to be 111.4 g C x m(-2). [ABSTRACT FROM AUTHOR]

The urban culture is a new culture in the modern urbanization advancement forms gradually, and the metropolitan culture is a national culture with highest level. Being one of the human most outstanding cultures, the scientific cultures appears with the scientific progress. Because of the restriction of traditional culture, our awareness about it is insufficient. We stress on the knowledge and the skill teaching excessively, so the scientific method and the spirit of science are quite defective. In our metropolitan culture, we should make good use of metropolitan scientific culture resources, and strengthen the building of scientific culture to promote the traditional culture of our country. [ABSTRACT FROM AUTHOR]

To investigate the effects of different initial processing methods on the quality of Fritillaria taipaiensis, this study explored the effects of anti-browning treatment, drying methods, and drying temperatures on the commercial characters, chromaticity values, and alkaloid and nucleoside components of Fritillariae Taipaiensis Bulbus. The results were comprehensively evaluated through correlation analysis(CA), principal component analysis(PCA), and hierarchical clustering analysis(HCA). Compared with those of the direct drying group(WD60), the chromaticity values(ΔE*) of the groups with scraped outer skin( FHB1) and mixed lime powder treatments(FHB2) were significantly reduced, indicating the inhibition of the browning process. The total alkaloid content of the group with mixed raw soil treatment(FHB3) and the FHB2 group showed no significant change, whereas that of the group with 5%Na Cl O solution rinse treatment(FHB4) was the lowest. Compared with air-blast dried(WD50) samples, the ΔE* values of freezedried(FS6) and vacuum-dried(FS5) samples were significantly decreased, with an increase in total alkaloid contents. Conversely,the ΔE* values of shade-dried(FS1) and sun-dried(FS2) samples were significantly increased, with severe browning and low total alkaloid contents. The total alkaloid contents of heat-pump-dried(FS4) samples showed no significant change, and their ΔE* value was significantly decreased, with a light degree of browning and favorable commercial characters. The total alkaloid content of air-blast dried samples initially increased and then decreased within the range of 40-80 ℃, and the highest content was recorded at 70 ℃. The ΔE* values of high-temperature air-blast dried samples(70-80 ℃) were smaller with a light degree of browning, whereas their texture was compact and lacked powder. CA revealed a significant relationship between the uracil content and chromaticity value of the samples(P&lt; 0. 05). The clustering relationships among samples subjected to different treatments were visualized via PCA and HCA. The results showed that FHB2 and air-blast drying(50-60 ℃) were more suitable for large-scale production, and heat pump drying could be a promising direction for future development. This study provides a scientific basis for optimizing the initial processing methods of Fritillaria taipaiensis.

This study aims to explore the effect of preventive administration of Yigong Powder on the learning and memory abilities of the mouse model of aging induced by D-galactose and decipher the underlying mechanism, so as to provide a basis for the application of Yigong Powder in the prevention and treatment of cognitive decline. Forty KM mice were randomized into control, model, donepezil(1.5 mg·kg~(-1)), and high-dose(7.5 g·kg~(-1)) and low-dose(3.75 g·kg~(-1)) Yigong Powder groups. The mice in other groups except the control group were injected with D-galactose(200 g·kg~(-1)) at the back of the neck for the modeling of aging. At the same time, the mice were administrated with corresponding drugs by gavage for one month. Morris water maze was used to examine the learning and memory abilities of the mice. Hematoxylin-eosin staining was employed to observe the pathological and morphological changes of the hippocampus. The immunofluorescence assay was employed to detect the expression of ionized calcium-binding adapter molecule 1(IBA1), glial fibrillary acidic protein(GFAP), chemokine C-X-C-motif ligand 12(CXCL12), chemokine C-X-C-motif receptor 4(CXCR4) in the hippocampus and observe the positional relationship between IBA1, GFAP, and CXCR4. Western blot was employed to determine the protein levels of extracellular regulated kinase(ERK), p-ERK, and tumor necrosis factor receptor 1(TNFR1). Enzyme-linked immunosorbent assay was employed to measure the levels of glutamate and tumor necrosis factor(TNF-α) in the brain tissue and the level of TNF-α in the serum and spleen. Yigong Powder significantly shortened the escape latency, increased the times crossing platforms, and prolonged the cumulative time in quadrants of the aging mice. It alleviated the nerve cell disarrangement, increased intercellular space, and cell degeneration or death in the hippocampus and reduced the pathology score of the damaged nerve. Moreover, Yigong Powder reduced the positive area of IBA1 and GFAP, reduced the levels of TNF-α in the brain tissue, serum, and spleen, and decreased spleen index. Furthermore, Yigong Powder decreased the average fluorescence intensity of CXCL12 and CXCR4, reduced CXCR4-positive astrocytes and microglia, down-regulated the protein levels of p-ERK/ERK and TNFR1, and lowered the level of glutamate in the brain tissue. This study showed that the preventive administration of Yigong Powder can ameliorate the learning and memory decline of the D-galactose-induced aging mice by regulating the immune function of the spleen and the CXCL12/CXCR4 signaling in the brain to reduce glutamate release. However, the mechanism of Yigong San in preventing and treating dementia via regulating spleen and stomach function remains to be studied.

The aim of this study is to explore differences in the peptidomics of Saccharomyces pastorianus protein hydrolysates treated with different enzymes. Briefly, differences in the peptide fingerprints and active peptides of neutral protease/papain-hydrolyzed S. pastorianus were analyzed using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) combined with PEAKS Online 1.7 analysis software, Peptide Ranker, and the BIOPEP database. Compared to traditional databases, the PEAKS Online uses de novo sequencing for analysis to obtain oligopeptides smaller than pentapeptides. It provides more comprehensive data of the peptide sample. In this study, enzymatic hydrolysates of S. pastorianus protein were prepared under the optimum conditions of neutral protease and papain respectively. In total, 7221 and 7062 polypeptides were identified in the hydrolysates of neutral protease and papain, respectively; among these polypeptides, 980 were common to the two enzymes. The 6241 and 6082 unique peptides found in the hydrolysates of neutral protease and papain, respectively, indicated that the peptide fingerprints of the two hydrolysates are quite different. Peptide Ranker predicted that 3013 (41.73%) and 3095 (43.83%) peptides were potentially bioactive in the hydrolysates of neutral protease and papain, respectively. According to the BIOPEP database, neutral protease and papain contained 295 and 357 active peptides, respectively; these peptides were mainly composed of angiotensin converting enzyme (ACE) inhibitors and dipeptidyl peptidase IV inhibitors and antioxidant peptides. The number of active peptides in the hydrolysate of papain was higher than that in the hydrolysate of neutral protease, but the total ion intensity of active peptides in the former was lower than that in the latter. This study revealed the influence of protease type on the composition of enzymatic hydrolysates from S. pastorianus protein. The above results provide a reference for the development of functional products of S. pastorianus protein peptides and the high-value utilization of yeast resources.

The phospholipid fatty acid (PLFA) technique was used to investigate the effects of fertilization on soil characteristics and microbial community of tomato in a solar greenhouse in Shouguang, Shandong Province, China, based on a long-term (12-year) fertilization experiment. The experiment involved a control (CK) and five fertilization treatments, namely, traditional nitrogen application (CN), traditional nitrogen application+straw return (CNS), optimized nitrogen application (SN), optimized nitrogen application+straw return (SNS), and organic manure nitrogen application+straw return (MNS). Results showed that the contents of soil organic matter and avai-lable P and K under all fertilization treatments were significantly higher than that of CK, but no significant difference among fertilization treatments. The traditional nitrogen application ( i.e. , CN and CNS) significantly decreased soil pH. The reduction of nitrogen fertilizer ( i.e. , SN, SNS and MNS) did not affect soil pH. Compared with CK, no straw treatment ( i.e. , CN and SN) and MNS did not affect the content of soil available N, but nitrogen combined with straw returned ( i.e. , CNS and SNS) significantly increased soil available N content. The optimized N with straw return ( i.e. , SNS) treatment resulted in the highest soil available N. Compared with CK and treatments without straw ( i.e. , CN, SN), treatments with straw ( i.e. , MNS, CNS and SNS) substantially changed soil microbial community structure and increased the biomass of soil bacteria, fungi, actinomycetes, mycorrhizal fungi, and the total amount of PLFA. SNS treatment had the highest soil microbial diversity, the highest biomass of soil bacteria, fungi, and actinomycetes, the highest total amount of PLFA, the highest ratios of fungi/bacteria and monounsaturated fatty acids/saturated fatty acids (MONO/SAT), the two indicators for soil ecosystem stability. Further, it had the lowest ratios of iso/anteiso fatty acids (i/a) and gram-positive/gram-negative bacteria (G + /G - ), the indicators for nutritional stress. Redundancy and correlation analysis revealed that soil organic matter was the main factor affecting soil microbial community structure, with a significant positive correlation with actinomycetes and gram-positive bacteria. In conclusion, straw returning combined with optimized nitrogen application ( i.e. , SNS, 8 t·hm -2 of wheat straw with N fertilizer reduced by 58.3%) could improve soil nutrient status, soil microbial biomass, soil microbial community structure, and soil ecological environment. It would be an effective measure for reducing fertilizer application and improving efficiency.

Objective: To evaluate safety and efficacy of autologous peripheral blood hematopoietic stem cell transplantation (auto-PBHSCT) with IBu precondition regimen consisting of high-dose idarubicin (IDA) and busulphan (Bu) for treatment of patients with low and intermediate risk acute myeloid leuekmia (AML)., Methods: A total of 11 patients with AML (5 low and 6 intermediate risk patients) treated with auto-PBHSCT with IBu precondition regimen (IDA 20 mg/m 2 , continuous i.v. from d-13 to d-11, Bu 0.8 mg/kg/q6h i.v. for 2h, from d-5 to d-2) from March 2011 to July 2014 were analyzed retrospectively. Adverse effects and transplantation-related mortality (TRM) were evaluated. Kaplan-Meier analysis was performed to calculate the overall survival (OS), disease-free survival (DFS) and cumulative relapse rate (RR). Cox regression was performed for univariate analysis for DFS., Results: Among the 11 patients, 10 patients obtained hematopoietic reconstitution, 1 patient died during transplantation, thus the TRM was 9.1%. The adverse effects were well tolerated. With median follow-up of 31.6 (8.7-52.5) months, 7 patients (63.3%) were alive, including 6 patients (54.5%) in continuous complete remission (CR). Median OS and DFS were not reached. The 3-year OS, DFS and RR were (57.7±16.3)%, (52.5±17.6)% and 47.5%, respectively. Univartiate analysis indicated that the age, sex, interval between diagnosis and transplantation, white blood cell count at diagnosis, risk-grouping (low or intermediate risk), disease status before transplantation (CR1 or CR2), and count of mononuclear cells for infusion all can not influence DFS(P>0.05, respectively)., Conclusion: The treatment of auto-PBHSCT with IBu precondition regimen for low to intermediate risk AML patients is safe and effective.

The genus Tripterygium is an immune suppressor in the Chinese traditional medicines. Due to the habitat destruction and anthropogenic over-exploitation, the wild genus Tripterygium plants have decreased dramatically in recent years or even been endangered. It is critical to evaluate and protect genus Tripterygium wild resource. In this research, simple sequence repeat (SSR) molecular markers were applied to the investigation of the genetic diversity and genetic structure of 28 populations for genus Tripterygium (396 samples from 9 provinces in China). We found a high level of genetic diversity (percentage of polymorphic loci PPL = 77.29%, Shannon’s information index I = 0.639 4; Nei’s expected heterozygosity H = 0.359 9) and high genetic differentiation among the populations (gene flow N&#95;m = 0.228 7). Based on Nei’s genetic distance, the phylogenic tree of populations was constructed and 28 populations were divided into 6 clusters according to STRUCTURE clustering analysis. T. hypoglaucumwas was mainly divided into 3 clusters, including Sichuan, Yunnan and Guizhou- Chongqing. T. regelii was separated to cluster 4, while T. wilfordii was divided into two clusters: the transition type LQ and NY were divided into cluster 5, and the others were in cluster 6. These results provide a theory basis for the conservation of wild resource, research of genetic polymorphism and molecular marker for assisted breeding of genus Tripterygium.

We studied the content of chemical compositions and correlation among species of Tripterygium genus by principal component analysis(PCA) and variance analysis(ANOVA), and we also studied the difference among the 3 species.Using [BMIm]PF6 ionic liquid-based ultrasonic-assisted extraction, we determined the contents of 11 compounds including wilforgine, wilforzine, triptophenolide, wilforine, triptoquinone A, triptolide, tripterin, egallocatechin, epigallocatechin, catechin, and epicatechin in 28 batches of the Tripterygium species by HPLC and PCA. Partial least squares analysis (PLS) and ANOVA were also performed to verify the results.The analysis results of PCA and PLS showed that three species of Tripterygium genus were clustered into three regions respectively, and triptoquinone A was the important factor which affected the aggregation of these three species.There was a significant difference among the contents of 11 chemical components in the three species(P<0.000 1).These results indicated that there was a certain correlation between the chemical compositions and the classification of the species, and the difference of the chemical compositions among the three species was obvious. In this work, the content determination method is rapid and accurate, and the analysis method is simple and convenient, which provides a reference for the classification, the efficacy and the toxicity of the species., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Ten compounds were isolated from Mylabris phalerata by using preparative HPLC and column chromatography over MCI gel. On the basis of physical-chemical properties, NMR and MS data analysis, the compounds were identified as 5'-[(1 R,2 R,3 S,6R)-1-hydroxymethyl-2-methyl-3,6-epoxycyclohexane-1,2-dicarboximide]- ethyl-2'-methyl-2'-butenoate (1),cantharidin (2), cyclo-(L-Pro-L-Ala) (3), cyclo-(R-Pro-R-Leu) (4), cyclo-(S-Pro-R-Leu) (5), cyclo-(D-Pro-L-Tyr) (6), indole-3-aldehyde (7), 3-indoleacetic acid (8), valerolactam (9), and 4-hydroxyphthalid (10).Compound 1 was a new compound, and compounds 2-10 were obtained from this genus for the first time. Compounds 1-9 were subjected to cytotoxic activity on HCT-116, HepG2, BGC-823, NCI-H1650, A2780 cell lines, and only compound 2 showed inhibitory effect on all cancer cell lines., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Using six kinds of ionic liquids as extractants, ultrasonic-assisted extraction coupled with HPLC method was developed for the simultaneous determination of wilforgine, wiforizine, triptophenolide, wilforine and triptoquinone A in Tripterygium hypoglaucum. The separation was performed on an Inertsil ODS-4 column with the mobile phase of acetonitrile-0.1% phosphoric acid in gradient elution at a flow rate of 0.75 mL•min⁻¹. Detection wavelength was 220 nm and the column temperature was 30℃. Under the optimal extractions, the results showed that triptophenolide and triptoquinone A had the highest extraction yield by using 0.6 mol•L⁻¹ [BMIm]PF6 methanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The calibration curves of triptophenolide and triptoquinone A showed a good linearity in the range of 0.000 65-0.026, 0.066 55-2.662 μg (r=0.999 9)respectively. The average recovery was 102.4% and 97.90% with RSD of 2.5% and 1.5%, respectively. Wilforgine, wiforizine and wilforine had the highest extraction yield when using 0.6 mol• L⁻¹ [BMIm]PF6absolute ethanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The content of wilforgine, wiforizine and wilforine from 0.023 9-0.956, 0.002 7-0.108, 0.006 4-0.256 μg showed a good linearity (r=0.999 9), and the average recovery was 100.6%,99.50% and 98.70% with RSD of 2.1%,1.9% and 2.7%, respectively. The results indicated that this method is convenient, reliable and green, and can be used as a reliableanalytical method for the quality control of T.hypoglaucum., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

This study was aimed to investigate the safety and effectiveness of tumor-ablative Chemotherapy combined with low intensity conditioning regiment BUCy/TBICy for patients with hematologic malignancies receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 30 patients with hematologic malignancies received above-mentioned therapeutic method from January 2012 to January 2013 was analyzed retrospectively, and the engraftment, GVHD, infection, conditioning-related toxicity, relapse and survival rates were evaluated. All the patients signed the informed consent before transplantation. The median follow-up duration was 20.5 (16.3-27.3) months. The results indicated that all the patients had been engrafted successfully. One year overall survival (OS) and disease-free survival (DFS) rates were 93.3% and 83.3% respectively. No conditioning-related toxicity occurred. The incidences of II-IV grade aGVHD was 37.9%, among which incidence of III-IV grade aGVHD was 3.4%; incidence of extensive cGVHD was 13.8%. So far, 1 case relapsed, 1 case displayed graft rejection, and poor function of graft occurred in 1 case, death occurred in 2 cases(6.7%). It is concluded that tumor-ablative chemotherapy combined with low intensity-modified BUCy/TBICy is safe and effective in allogeneic hematopoietic stem cell transplantation for hematologic malignancies, and it is useful to reduce relapse of hematologic malignancies after transplantation.

In this study, a large area of well preserved Zostera japonica dominated meadow was found in a coastal lagoon, Swan Lake, in Rongcheng of Shandong Province. Due to its unique geographical position and high biomass, this meadow may act as a typical Z. japonica bed in the coastal area of Shandong. From September, 2011 to October, 2012, an annual investigation was conducted on the Z. japonica and its habitats in east coast of Swan Lake, and the distribution of the Z. japonica and its habitats ecological characteristics were preliminarily understood. The major ingredients of sediments particles in the Z. japonica bed was sand (81%) and silt (14%). The C and N contents in the sediments were the highest in winter, and the C/N ratio was the highest in autumn. The shoot density, shoot height, and biomass of the Z. japonica were all significantly correlated with water temperature (P < 0.05). There was an obvious change in the Z. japonica growth among seasons, with the peak biomass obtained in August-September. The C and N contents and C/N in Z. japonica leaves also varied with seasons. The leaf C content was significantly higher in autumn than in spring and summer (P < 0.05), the leaf N content was significantly lower in summer than in spring and autumn (P < 0.01), whereas the leaf C/N ratio was significantly higher in summer than in spring (P < 0.05). The annual carbon sequestration by the Z. japonica in the Swan Lake was estimated to be 111.4 g C x m(-2).

Objective: To explore the expression and clinical significance of T cell immunoglobulin mucin (TIM)-1, TIM-3 and T cell-specific transcription factors T-bet and GATA-3 in spleen mononuclear cells in patients with primary immune thrombocytopenia (ITP)., Methods: The spleen samples were obtained from 17 active ITP patients and 10 controls with spleen traumatic rupture. By using real-time quantitative polymerase chain reaction, the mRNA expressions of TIM-3, TIM1, T-bet and GATA-3 were studied in all subjects., Results: TIM-3 mRNA levels of active ITP patients were significantly decreased to (29 ± 16)% of that of control, TIM-1 mRNA levels of active ITP patients increased to (3.20 ± 2.18) folds of that of control, but the difference was not significant. The ratio of TIM-1/ TIM-3 was elevated in active ITP patients. T-bet mRNA levels were up-regulated in ITP patients by (2.82 ± 1.57) folds (P<0.05) and the expression of GATA3 was decreased by 14% folds (P<0.05) compared to controls. The ratio of T-bet/GATA3 were significantly elevated in ITP patients., Conclusion: The imbalance between TIM-3 and TIM-1 expression might play an important role in pathogenesis of ITP.

Objective: To investigate clinical characteristics and mutation of the LKB1 gene in a Peutz-Jeghers syndrome (PJS) pedigree., Method: Clinical data of a PJS family were analyzed and LKB1 gene mutation was detected by systematic screening with multiplex ligation-dependent probe amplification (MLPA) and DNA sequencing. Meanwhile, two hundred and fifty healthy adults were enrolled in this study and denaturing high performance liquid chromatography (PCR-DHPLC) was carried out to verify the mutation excluding polymorphism sites found in this family. Changes in protein structure and function caused by the mutated coding sequence was analyzed by SWISS-MODEL software., Result: The proband had pigmented mucocutaneous lesions and multiple hamartomatous polyps in the gastrointestinal tract. There was no fragment deletion of LKB1 gene detected by MLPA. Among PJS family and 250 healthy adults, germline mutation c. 924G > C of LKB1 which cause Trp308Cys in protein sequence was identified only in the proband and another affected member. LKB1 protein activity could be reduced due to changes in LKB1 protein conformation structure by Trp308Cys., Conclusion: Peutz-Jeghers syndrome (PJS) is an autosomal dominant disorder characterised by mucocutaneous pigmentation, multiple gastrointestinal hamartomatous polyps and heredofamilial nature. Gene identification and mutagen screening of LKB1 gene in all PJS patients and first degree relatives will contribute to a definite diagnosis and improve the life span of the family.

Objective: To screen for potential mutations of LKB1 gene in Chinese familial Peutz-Jeghers syndrome (PJS) patients and analyze their clinical manifestations., Methods: Eleven PJS families were collected and genomic DNA of peripheral blood was extracted. Typically mucosal pigmentation and hamartomatous polyps were present in all 11 probands. Mutation screening of the probands were carried out by PCR and direct sequencing. Two hundred and fifty healthy adults were enrolled as normal controls, for whom genomic DNA of peripheral blood was also extracted. PCR-denaturing high performance liquid chromatography was carried out to verify the mutation identified in the patients., Results: Nine germline mutations were identified in eight PJS patients, which included 7 point mutations, 1 deletion and 1 insertion. Among these, 4 were considered to be pathogenic, of which 2 were de novel, 4 were considered to be polymorphism, and 1 was uncertain., Conclusion: LKB1 gene mutations with pathogenic effect are a common cause of familial PJS in Chinese patients. Most mutations are point mutations.

Spectral difference is an important aspect for extracting shadows of buildings. Based on the analysis of the relationship between the building heights and the shadows in ALOS Images, the paper presents the principle and the method for building heights estimation in a city from the shadows of an image, and works out a feasible approach to determining shadow zones in a panchromatic ALOS Images. It has also contributed a standard process for extracting buildings distribution information of different heights in a city from the shadows in a panchromatic ALOS Images. A result with about 87.6% accuracy has been achieved while applying this technique to Tianjin City, which has demonstrated prospective applications of satellite remote sensing to urban purposes.

Collecting information from the database of China National Knowledge Information (CNKI) and VIP database of Chinese Journals, an overall analysis and review on treatment of oculomotor paralysis with electroacupuncture in recently years are made in the paper. The electroacupuncture, which can provide constant stimulation and reinforce the effect of acupuncture, is the major therapy to treat oculomotor paralysis. However, the best stimulate threshold of electroacupuncture on oculorotary muscles has not been reported yet. The parameters of electroacupuncture adopted in clinic are not standardized, which lead to unfavorable therapeutic effect as well as unclear mechanism of treatment. Therefore, it is suggested that the study should focus on enhancing the research level and broadening researcher's mind to explore the best parameter for the electroacupuncture stimulation on the oculorotary muscles, to make clear the mechanism of treatment and to search for valuable observation indicators in the future.

Dye analysis is important to the understanding of fabric color degradation and technical development of ancient printing and dyeing. In the present study, thin layer chromatography and Raman spectroscopy were used for the analysis of blue dyes from 6 silk fabric of Tang dynasty and decorative painting of Jian Fu Gong, Forbidden City. The applicability of these two methods in the cultural heritages was also studied. The results indicate that all these blue substances are indigo; indigo was not only used as dye in ancient fabrics, but also as pigment in decorative painting of historic building, so it is used widely. Both analytic methods have advantages and disadvantages; Raman spectroscopy is nondestructive analysis; thin layer chromatography needs small amount of sample, but could give more information.

Objective: To investigate the association of the single-nucleotide polymorphism (SNP) IVS10+12 G>A in hMSH2 gene with colorectal cancer in a Chinese population of Jiangsu province., Methods: A case-control study to investigate whether this SNP affects the risk of developing colorectal cancer was conducted. Subjects included 108 colorectal cancer patients and 180 healthy individuals. Peripheral white blood cell DNA was obtained from all subjects. The hMSH2 gene IVS10+12 G>A was genotyped using a PCR-based DHPLC, the existence of IVS10+12 G>A was verified by DNA sequencing., Results: The allele frequency of the IVS10+12 G>A in the hMSH2 gene in the healthy individuals was 51.7%. There was significant difference in the frequency of the IVS10+12 G>A between patients and healthy controls (P<0.05), and between familial patients and healthy controls (P<0.05). There was also significant difference of the frequency of the IVS10+12 G>A between patients younger than 50 years, and patients with high consumption of fried food and pickled vegetable and healthy controls respectively (P<0.05)., Conclusion: This SNP may be associated with colorectal cancers in Chinese. Further investigation with larger sample size is needed.

Objective: To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses., Methods: Obtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance., Results: Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity., Conclusion: Successfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.

Objective: To investigate the etiological role of hMLH1 gene A655 polymorphism in colorectal cancer., Methods: A case-control study was carried out, including 115 colorectal cancer patients and 135 healthy people as control. Genomic DNA was extracted from peripheral white blood cell from all the subjects. Polymorphism was detected by PCR-based DHPLC analysis and verified by DNA sequencing., Results: The hMLH1 gene A655G polymorphism was detected in 3.0% of healthy people and 11.3% of colorectal cancer patients (P<0.01), and the difference was significant (P<0.01). The hMLH1 gene A655G polymorphism was detected in 8.2% of tubular adenocarcinoma or tubular-papillary adenocarcinoma and 27.8% of mucinous adenocarcinoma, which was also significant (P<0.05).Meanwhile, hMLH1 gene A655G polymorphism was not associated with age, gender and lymphatic metastasis (all P>0.05)., Conclusions: The hMLH1 gene A655G polymorphism may play a role in the pathogenesis of colorectal cancer. Determination of the polymorphism may be a potential marker to predict the prognosis of colorectal cancer patients.

AIDS caused by HIV-1, is a major threat to human being. An anti-HIV formulation from Chinese herbs, so called "Qu Du Zeng Ning", have been recently developed. In this work, the pharmacodynamics of the formulation in vitro was studied. The results showed that Qu Du Zeng Ning inhibit the replication of HIV-1 efficiently in all cell-based assay, with IC50 at 105.2, 70.7, 77.4 microg mL(-1), separately. A significant synergy between the formulation and zidovudine (AZT) was observed, and it also showed a potent activity against HIV-1 drug-resistant mutant.

Objective: To express and purify H5N1 influenza virus (A/Anhui/1/2005) NP in prokaryotic system and to explore the NP-interacting proteins of human bronchial epithelial cells BEAS-2B in vitro., Methods: The full length H5N1 NP gene fragment was amplified by PCR, inserted into prokaryotic expression vector (pET30a) to generate NP expression plasmid pET30a-NP. After transforming pET30a-NP into E. coli (BL21), the expression of soluble NP protein was induced by IPTG. The expressed NP protein was purified by two steps with metal chelation chromatography and ion exchange chromatography. Then the total proteins of BEAS-2B cells was extracted for screening the components which have protein-protein interaction with purified NP by pull-down and LC-MS/MS methods., Results: The expression of H5N1 NP protein could be induced by IPTG in bacterial system using expression plasmid pET30a-NP. The soluble NP was purified. Twenty proteins were found by pull-down and LC-MS/MS, the further experiments may be needed to prove protein-protein interaction between them., Conclusion: The soluble H5N1 NP fusion protein with high purity was obtained and twenty proteins were found which could interact with it by pull-down and LC-MS/MS.

Objective: To Construction of P and NP genes eukaryotic expression vectors of Newcastle Disease Virus LaSota strain,study its reverse genetics and functional genome of NDV., Methods: P, NP genes were amplified and cloned into pGEM-T easy vector and then subcloned into pcDNA3.1 (+) expression vector respectively, the recombinant plasmids were named pcDNA3.1 (+)-P and pcDNA3.1 (+)-NP, Recombinant plasmids were transfected into 293 and BHK-21 cells respectively and were detected using IE and Western blot analysis., Results: Expression of P, NP genes were detected and confirmed by the IE and WB analysis., Conclusion: The recombinant eukaryotic plasmids pcDNA3. 1(+)-P, pcDNA3.1 (+)-NP were expressed in 293 and BHK-21 cells successfully. This research may be helpful for further study of reverse genetics and functional genome of NDV.

The present paper did some researches on the deterioration mechanism and the changes in crystallinity of silk fabrics by means of the X-ray diffraction analysis. The samples artificially aged and excavated from Hubei, Innermongolia, Shaanxi and Qinghai provinces were analyzed. The artificial aging was done by simulating three main natural aging factors: light, heat and hydrolysis. The analytical results show that X-ray diffraction analysis could reveal the aging process and characteristic of silk, as well as the changes in crystallinity during silk aging. The X-ray diffraction analysis is of practical value for the conservation state and aging mechanism studies of ancient silk. In addition, X-ray diffraction analysis could also provide important information on ancient technology of textile and apparel.

Objective: To investigate immunity of a recombinant adenovirus vaccine (rAdV) containing codon-modified neuraminidase (Mod. NA) gene of H5N1 influenza virus in BALB/c mice and to screen for appropriate dose., Methods: BALB/c mice were immunized with the rAdV-Mod.NA vaccine intramuscularly twice (double injection at 0 and 4th week) in three groups, low dosage (10(5) TCID50 per dose), medium dosage (10(7) TCID50 per dose) and high dosage (10(9) TCID50 per dose). The effect of humoral and cell-mediated immunity were analysed at 5th week., Results: (1) The rAdV-Mod.NA vaccine could elicit both humoral and cell-mediated robust NA specific immunity in mice by neuraminidase inhibitor assay and IFN-gamma ELISpot assay; (2) 10(7) TCID50 per dose was the appropriate dose; (3) Peptide NA(109-124): CRTFFLTQGALLNDKH and peptide NA(182-199): AVAVLKYNGIITDTIKSW were the dominant epitopes for neuraminidase-immunized BALB/c mice, which was screened out from the whole length of neuraminidase of an H5N1 virus, A/Anhui/1l/2005., Conclusion: The recombinant adenovirus NA could induce specific humoral and cellular immune responses in BALB/c after immunization, which suggest rAdV-Mod.NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.

The aim of this study is to develop the recombinant adenovirus vaccine (rAdV) candidates containing neuraminidase (NA) gene of H5N1 influenza virus and test in BALB/c mice the effect of cell-mediated immunity. In this study, two kind of NA gene (WtNA gene, the wild type; Mod. NA gene, the codon-modified type) derived from H5N1 influenza virus (A/Anhui/1/2005) were cloned and inserted respectively into plasmid of adenovirus vector, then the rAdV vaccines candidates (rAdV-WtNA and rAdV-Mod. NA) were developed and purified, followed by immunization intramuscularly (10(9) TCID50 per dose, double injection at 0 and 4th week) in BALB/c mice, the effect of cell-mediated immunity were analysed at 5th week. Results indicated that: (i) NA protein expression was detected in two rAdV vaccines candidates by Western blotting; (ii) the rAdV-Mod. NA vaccine could elicit more robust NA specific cell-mediated immunity in mice than that of rAdV-WtNA vaccine (P = 0. 016) by IFN-gamma ELIspot assay. These findings suggested rAdV-Mod. NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.

The aim of study was to establish the packaging system of the recombinant lentiviral vector encoding Gfi1 gene for eukaryotic expression and to realize the efficient, stable expression of Gfi1 32D cells so as to provide effective platform for further studying the development of Gfi1 gene in hematologic malignancies. The three-plasmid recombinant lentiviral vector consisting of transfer plasmid (pLOX-Gfi1/pLOX), the packaging plasmid (pCMVDeltaR8.2) and the envelop plasmid (pMD.G) was prepared and purified. Human embryonic kidney 293T cells were cotransfected with the three plasmids by lipofectamine 2000. After transfection for 48 hours, the viral supernatant was collected and the target cell 32D was transfected with the recombinant lentivirus; the Gfi1 integration and expression in 293T and 32D cells were detected by Western-blot. The results showed that the three plasmids of lentivirus could be transfected into 293T with high efficiency and packaged successfully, and the Gfi1 protein could be detected by fluorescent microscopy. The recombinant lentiviruses carrying Gfi1 could transfer and deliver Gfi1 gene to 32D cells, and the Gfi1 expression in 293T and 32D cell could be detected by Western blot. It is concluded that the recombinant lentivirus carrying Gfi1 can deliver target gene to 32D cells with high efficiency, and the expression of Gfi1 protein is stable in 32D.

Objective: To explore the feasibility and safety of conditioning regimen containing fludarabine (Flud) for haploidentical hematopoietic stem cell transplantation (HSCT)., Methods: Preparative regimen containing Flud 40 mgxm(-2)xd(-1) on day -7 to -3 in place of cyclophosphamide (CTX) for haploidentical HSCT was given to 35 patients with hematologic malignancies (4 standard risk, 16 high risk, 15 relapse with no remission). All donors received rhG-CSF followed by HSC harvest. One patient received peripheral blood HSCT (PBSCT), one bone marrow transplantation (BMT), and the others BM combination with PBSCT. The regimen-associated side effect, engraftment, incidence of graft-versus-host disease (GVHD) and disease-free survival (DFS) probabilities were observed., Results: All patients achieved sustained, full donor-type engraftment. Thirty-four patients obtained primary durable engraftment, and 1 who rejected graft from his mother obtained successful durable engraftment after the second graft from his father. The cumulative incidence of grade III-IV acute GVHD and chronic GVHD was 12.1% and 31.7%, respectively. With a follow-up duration of 8-25 months, 6 patients were dead, in which 3 died of relapse, 2 of acute GVHD, 1 of fungal infection, none died of regimen-associated side effect. The other 29 patients remained alive and DFS probability was 79.7%., Conclusion: Flud based conditioning regimens for haploidentical HSCT is safe and feasible, which reduces regimen-associated side effect, with no increasing the rate of relapse and infection, and decreases the incidence of aGVHD.

Objective: To investigate the southern region of Zhejiang hepatitis E virus (HEV) infection., Methods: A cluster sampling strategy was used to sample all blood donors from February to October in 2008 in Wenzhou blood center. Their blood was tested for IgG and IgM antibody against HEV. Reverse transcriptase-polymerase chain reaction(RT-PCR) and sequencing were applied to detect its genotype and sequence homology in HEV IgM-positive specimen., Results: The prevalence of anti-HEV IgG in 3044 cases of blood donors was 33.28%. IgG increased with age. There are certain increase in positive rates between the 20-year-old group and over 40 years of age group from 21.16% to 50.36%. The positive rate of IgM was 0.92%. The ratio of infection among different age group was the highest in the age range from 31 to 40 years and up to 1.90%. IgG and IgM through their negative and positive analysis of samples found in their group with donors age, sex and blood type does not significantly related to each other. Nucleic acids were found in three cases through PCR amplification in all 28 cases of HEV IgM positive samples. The total positive rate was one-thousandth, of which two cases for gene 4, 1 cases of infection for gene 1., Conclusion: The results indicate that there was a certain percentage of HEV virus in voluntary blood donors in south Zhejiang.

Objective: To construct adenovirus vector vaccine against H5N1 influenza virus and study on the immunogenicity., Methods: In this study, we amplified hemagglutinin (HA) gene sequence of H5N1 influenza virus (A/Anhui/1/2005), then constructed an adenovirus vector vaccine (Adv-HA), followed by tests in BALB/c mice for the immunogenicity with the vaccine and immunization strategies., Results: The recombinate Adv-HA vaccine could effectively induce both humoral and cellular immunity against human H5N1 influenza virus., Conclusion: The Adv-HA vaccination against H5N1 influenza is a potential strategy and worthy of further investigation.

Objective: To prepare HIV-1 subtype C Gp120 protein and to produce its polyclonal antibodies., Methods: A C-terminal fragment of gp120 gene was amplified by PCR from a plasmid expressing full-length HIV-1 subtype C gp160 gene. The length of the subtype C gp120 fragment was 612 nt and it encodes 204 amino acid residues. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-30a) and recombinant pET-30a-gp120 was expressed in Escherichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine (His6) tag at the C-terminus for convenient purification. To produce subtype C Gp120-specific polyclonal antibodies, New-Zealand rabbit was immunized with the purified Gp120 protein. Serum samples were tested by enzyme-linked immunosorbent assays (ELISA) to determine the level of antibodies. And Western blotting was used to further verify whether the polyclonal antibodies could specifically recognize subtype C Gp160 protein expressed in mammalian cells., Results: HIV-1 subtype C Gp120 protein was successfully acquired and the titer of its polyclonal antibodies was 1:204 800. The polyclonal antibodies efficiently recognized Subtype C Gp160 protein expressed in COS-1 cells., Conclusion: HIV-1 subtype C Gp120 fusion protein with high purity was obtained and its corresponding polyclonal antibodies with high titer were produced.

Objective: To establish a specific and sensitive Enzyme-linked immunosorbent Assay (ELISA) kit for detection of HIV-1 antibody in urine using Escherichia coli expression products as coating antigen., Methods: The truncated HIV-1 gp41 gene fragment of major antigenic epitopes was inserted into the plasmid pET22b to obtain expression plasmid pET22b-mgp41. The recombinant antigen was expressed in BL21 (DE3) strains of Escherichia coli and was purified by immobilized metal chelation and gel filtration chromatography. Using this antigen as coating antigen, a HIV-1 urine antibody ELISA kit was developed. In order to examine the clinical utility of the kit, 5437 urine samples were assayed, which consisted of 641 urine samples from HIV infected patients and 4796 samples from normal subjects. Results The purity of purified antigen is up to 95%. Anti-HIV antibodies were detected in all the urine samples from HIV infected patients, and the diagnostic sensitivity for HIV-1 infection was 100%. In healthy control group, 71 cases showed false positive, the specificity was 98.52%., Conclusion: The HIV-1 urine antibody kit can be used in screening and diagnosing for HIV-1 infection.

Objective: To investigate the status of hypermethylation in the promoter 1A region of the adenomatus polyposis coli (APC) gene in 3 familial adenomatous polyposis (FAP) pedigrees and to screen large fragment deletions in the APC gene., Methods: DNA from tumor tissues and corresponding normal tissues of 5 FAP patients was modified by sodium bisulfite. Then the methylation status of the APC gene was analyzed by methylation specific-PCR (MSP) and DNA sequencing. Multiplex ligation-dependent probe amplification (MLPA) was used to screen aberrations involving large fragments from all the 15 exons and promoter region of APC gene., Results: No methylation was present in normal tissues. Hypermethylation was found in tumor tissues of one proband and her son. Loss of heterozygosity was observed in another patient from the same FAP family., Conclusion: Aberrant methylation of the APC promoter region provides an important mechanism for impairing APC function and may occur early during colon neoplasia progression. Loss of heterozygosity may play a role in patients with classical polyposis.

Objective: To observe the effects of Feixian Formula, a compound traditional Chinese herbal medicine for treating pulmonary fibrosis, on bleomycin-induced pulmonary fibrosis in rats, and its influence on serum transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor (PDGF)., Methods: Seventy-two male Wistar rats were infused with bleomycin (1 mg/kg) through tracheal intubation to induce pulmonary fibrosis, and they were randomly divided into untreated group (n=24), prednisone-treated group (n=24) and Feixian Formula-treated group (n=24). Fifteen male Wistar rats of the sham-operated group were infused with equivalent normal saline. Twenty-four hours after operation, prednisone (5 mg/kg) and Feixian Formula (1.25 g/kg) were given to the prednisone-treated group and Feixian Formula-treated group respectively by intragastric administration once a day. Equivalent saline was administered to rats of the untreated group and sham-operated group. On the 14th, 28th and 45th day, 5 rats in the sham-operated group and 8 rats in each of the other three groups were dissected to observe pathologic changes of the lung tissues, and the levels of serum TGF-beta1 and PDGF were determined by enzyme-linked immunosorbent assay., Results: At the 45th day, the degree of pulmonary interstitial fibrosis was lesser in rats of the Feixian Formula-treated group as compared with those of the untreated group and prednisone-treated group. The levels of serum TGF-beta1 and PDGF were increased, and were significantly higher than those of the sham-operated group, especially on the 45th day (P<0.05). Changes of TGF-beta1 level in the prednisone-treated group and the Feixian Formula-treated group were similar to untreated group (P>0.05), and there was no significant difference between the prednisone-treated group and the Feixian Formula-treated group (P>0.05). PDGF in the Feixian Formula-treated group reached the highest level on the 14th day, significantly higher than those of the other three groups (P<0.01). Then it decreased, and was close to that of the sham-operated group on the 45th day (P=0.792). The levels of PDGF in untreated group and prednisone-treated group were increased depending on time, and were obviously higher than that of the sham-operated group on the 45th day (P<0.01)., Conclusion: Feixian Formula can relieve bleomycin-induced pulmonary fibrosis in rats, and the mechanism of its action may be related to down-regulating serum PDGF.

In order to improve the expression of human avian influenza virus hemagglutinin (HA) and meet the pandemic influenza vaccine needs, we optimized and synthesized the whole length of HA gene of H5N1 (A/Anhui/1/2005) influenza virus in accordance with the human's codon preference, and inserted it to the eukaryotic expression vector pDC315 to construct a eukaryotic expression vector pDC315-Mod. HA. This plasmid and the eukaryotic expression vector pDC315-Wt. HA containing wild HA gene were transfected into 293T cells respectively to compare the expression of HA protein. The results showed that according to the comparison and identification by indirect immunofluorescence assay and Western blot test, the expression of HA protein in 293T cells was significantly improved after codon optimization. This laid a foundation for pandemic influenza vaccine research.

Objective: To explore the characteristics of adenomatous polyposis coli (APC) gene germline mutations in Chinese patients with familial adenomatous polyposis (FAP)., Methods: Eighteen members from nine FAP pedigrees were studied by using multiplex ligation-dependent probe amplification(MLPA) to detect large fragment deletion of APC gene. Then, PCR were performed to amplify all exons of APC gene for mutation screening by denaturing high performance liquid chromatography (DHPLC). When abnormal elution profile of DHPLC was found, DNA sequencing was performed to determine the mutations., Results: Mutations were identified in three pedigrees among the nine pedigrees. They were c.3184&#95;3187 del CAAA in pedigree 2, c.5432C to T in pedigree 4 and c.3925&#95;3929 del AAAAG in pedigree 9 respectively. Among them, c.5432C to T was novel., Conclusion: APC gene germline mutations can cause the development of FAP. The FAP patients without APC gene germline mutations could be caused by other mechanisms.

Objective: To study the approach for developing a rat model of pulmonary fibrosis induced by bleomycin (BLM)., Method: Different doses (7, 6, 5, 3.4, 2, 1 mg/kg) of bleomycin A5-saline were infused into the rats' lung in bleomycin-treated group through tracheal intubation, and rats in sham-operated group were infused with same volume of saline. The living state and lung pathology of the rats were observed. The author deeply studied the condition of the rats in 1 mg/kg bleomycin-treated group, and the changes of body weight and lung pathology were observed. Lung quotient, the content of transforming growth factor beta1(TGF-beta1) and platelet-derived growth factor (PDGF) in serum were measured on the 14th, 28th and 45th day of the experiment., Results: The study demonstrated that infusing large doses of bleomycin A5 quickly through tracheal intubation had a high mortality, and infusing 1 mg/kg quickly could successfully develop an animal model of pulmonary fibrosis. Compared with the sham-operated group, fibrosis was appeared obviously in the rats' lung in 1 mg/kg bleomycin A5-treated group after 14 days of experiment, diffuse fibrosis was appeared after 28 days of experiment, and the fibrosis became more severe after 45 days of experiment. The body weight of the rats in bleomycin-treated group was declined after 3, 7 and 14 days of experiment as compared with the sham-operated group (P<0.01). Twenty-one days after the experiment, the body weight was declined too, but there was no significant difference between the bleomycin-treated group and the sham-operated group (P>0.05). Lung quotient was increased 14, 28 and 45 days after the experiment (P<0.01), the level of serum TGF-beta1 began to increase since 28 days after the experiment (P<0.05, P<0.01), and the level of serum PDGF also increased gradually 45 days after the experiment (P<0.05). And the mortality rate of 1 mg/kg bleomycin A5-treated group was lower than those of the other doses of bleomycin A5-treated groups., Conclusion: A rat model of pulmonary fibrosis can be duplicated successfully by infusing 1 mg/kg bleomycin A5 quickly through tracheal intubation.

The dhaB gene encoding glycerol dehydratase and dhaG dhaF gene encoding glycerol dehydratase reactivating factor from Citrobacterfreundii were amplified by PCR. The temperature control expression vector pHsh harboring yqhD, dhaB, dhaG and dhaF gene was transformed into E. coli JM109 to yield the recombinant strain E. coli JM109 (pHsh-dhaB-dhaG-dhaF-yqhD). The results from SDS-PAGE analysis show that the recombinant product was consistent with the molecular weight predicted from gene sequence. The fermentation result show that the yield of 1,3-propanediol was increased by 28% compared with E. coli JM109(pHsh-dhaB-yqhD).

Objective: To investigate the effect of dongchong xiacao capsule on the airway inflammation of asthmatic patients and to explore the relevant mechanism of therapeutic effect of Dongchongxiacao capsule., Method: Sixty patients with moderate persistent asthma were randomized into the treatment group (n=30) and the control group (n=30). Inhaled corticosteroid and as-needed beta-agonist were used in the treatment group while this therapy plus dongchong xiacao capsule were used in the control group for two months. Serum IL-4, IFN-gamma, sICAM-1, MMP-9, IgG, IgE level were assessed at randomization and 2 months after randomization., Result: The serum level of IgE, sICAM-1, IL-4 and MMP-9 of the treatment group was lowered to a greater degree than that of the control group (P < 0.05 or P < 0.01 )., Conclusion: Dongchong xiacao capsule can reduce the serum markers of airway inflammation, which suggests this therapy bares the anti-inflammation effects probably through regulating the balance of TH1/TH2, inhibiting the activity of adherence molecule and reducing IgE production. It may also have the effect of reversing airway remodeling, which needs further research to determine.

Objective: To observe the therapeutic effect of eye-needling combined with medication for treatment of ophthalmoplegia and explore the possible mechanism., Methods: One hundred and twenty cases were randomly divided into a treatment group and a control group. According to etiological factors, the control group were treated with medication and the treatment group with the medication plus eye-acupuncture at main point ocular muscles. Changes of the rima oculi, the range of ocular movement and the dialopia angle after treatment were recorded and statistically analyzed in the two groups., Results: The total effective rate was 93.4% and the cured rate was 54.1% in the treatment group, and 74.6% and 18.6% in the control group, with significant difference between the two groups (P < 0. 01)., Conclusion: Eye-needling combined with medication has an obvious therapeutic effect which is better than simple medication for ophthalmoplegia.

Polymorphism which is either in the thymidylate synthase (TS)enhancer region (TSER) of the 5-primer untranslation region (5' UTR) or in the 3-primer untranslation region (3' UTR) has been reported to be associated with the alterations in TS mRNA and protein levels. The TSER is characteristic of the presence of variable double (2R) and triple (3R) number tandem repeats (VNTRs). In addition to VNTRs, single nucleotide polymorphism (SNPs) in 3R, as well as the polymorphism of the fragment length(FLP) in the TS 3'-UTR which is characteristic of the presence or absence of a 6 bp- nucleotide fragment, has recently been reported to be associated with the response to 5-fuorouracil (5-FU)-based chemo-therapy. The aim of the present study was to develop a specifc denaturing high-performance liquid chromatography (DHPLC) method for the rapid and simultaneous detection of these variations in clinical samples. Multi-PCR primers were designed to amplify the two regions simultaneously, The 8.6 min DHPLC gradient was optimized to include the analysis of multiplexed TSER/3' UTR chromatogram peaks, allowing for the simultaneous detection of 28 bp VNTRs and 6 bp FLP under nondenaturing conditions (50). The optimal melting temperature was determined experimentally for the detection of SNP in the TSER VNTRs. Finally, the DHPLC analysis was verified in parallel with PCR-RFLP and sequencing. The optimized DHPLC method resolved 100% of the known TS variations, discriminated between homozygous and heterozy-gous genotypes. This developed DHPLC method could permit the rapid, sensitive, and accurate identification of the TS genotypes (VNTRs, SNPs, and FLP).

Objective: To study the immune effect of a chimeric adenovirus type 5 vector with type 35 fiber (rAd5/F35) vaccine in BALB/c mice., Methods: The expression of HIV Gag protein was determined using indirect immunofluorescent staining. The rAd5/F35-mod.gag vector was injected intramuscularly to mice. The IgG antibody was detected by ELISA and CTL response was detected by intracellular cytokine stain assay., Results: The rAd5/F35-mod.gag vector could express HIV Gag protein in vitro and generate strong HIV-specific immune responses in vivo. But anti-Ad5 immunity could limit its immunogenicity in vivo., Conclusion: The rAd5/F35-mod.gag vector can elicit specific CTL response and IgG antibody in animal model. In mice with high Ad5 vector-specific immunity, Ad5/F35-mod.gag showed lower level of Gag specific CTL and antibody response than in mice without pre-existing adenovirus type 5 immunity. The results indicated that fiber exchange alone does not evade pre-existing Ad5 immunity.

Objective: To study the HIV related high-risk behaviors and associated factors on the spread of HIV among men having sex with men(MSM) who lived in mainland China and to provide evidence for developing related policies and intervention measures., Methods: Questionnaires were distributed at gay bars and volunteer activity venues in six big cities of China. Data on 1389 valid cases was collected and urine HIV screening test was provided. Data was analyzed with SPSS 11.0., Results: The respondents were 27.62 year olds on average with an average age for first intercourse at 19.18. The most commonly available way of finding a sex partner was through internet(43.07%), followed by gay bar and public bathrooms(35.29 % ). 6 months prior to the study, the average number of their male sex partners was 5.69 including 4.37 unfamiliar sex partners and the average number of anal-intercourse was 4.33 with 11.61 per cent of them had experienced group sex. 13 cases of them showed positive results for preliminary urine HIV screening test. In the prior 6 months, 32.46 per cent of those who had experienced intercourse using condom every time while 76.37 per cent of them during the last sex episode. In the previous 6 months, 47.18 per cent of those who had experienced intercourse with women never used condoms., Conclusion: HIV high-risk behaviors are ubiquitous among MSM and AIDS intervention measures should be significantly strengthened in reaching MSM via a wide variety of conduits, especially internet. Meanwhile, a gay-friendly environment for prevention and control of AIDS is vital.

Background: To develop an ELISA method using Herpesvirus hominis type 4 (EBV) IgG/Zebra as capture antigen for large population screening., Methods: The ELISA method used purified ZEBRA antigen to detect the IgG/ZEBRA antibody from serum in nasopharyngeal carcinoma (NPC) and normal healthy subjects., Results: Of 288 NPC sera, 262 were detected positive, the sensitivity was 91%, while 5 of 96 normal sera were detected positive, the specificity was 94.8% and the results of NPC group and healthy group displayed significant difference (P less than 0.001). IgA/VCA, IgA/EA, IgG/EA in immunoenzyme methods and ZEBRA ELISA were compared during the NPC screening in two cities: Huizhou, Guangdong and Guiping, Guangxi, 5463 and 2017 samples respectively were tested and 5 earlier NPC patients were found., Conclusion: The results indicate that this method has high specificity and sensitivity, and can be used for large population screening to assist early phase NPC diagnosis.

Objective: To detect the adenomatous polyposis coli (APC) gene germline mutation in the proband and her family members with familial adenomatous polyposis (FAP)., Methods: The diagnosis of a patient with FAP was validated by colonoscopy, pathology and the family history. The systematic screening with multiplex ligation-dependent probe amplification (MLPA), denaturing high-performance liquid chromatography (DHPLC) and DNA sequencing were carried out to detect APC gene germline mutations., Results: A novel mutation c.1999 C >T (Q667X) of APC, which leads to premature termination of the protein, was identified in this family. This mutation manifested an aggressive form of FAP with early onset of colorectal adenocarcinoma and colonic adenoma., Conclusion: The mutation of APC Q667X is the cause of clinical phenotype of this family with FAP, and the prophylactic colectomy for the affected family members should be considered.