The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

Objective To analyze the change in infrared(IR)spectral information and to screen out the classification of major compounds affecting information difference in IR spectra of Trichosanthes before and after steaming. Methods The similarity between the original IR fingerprint and the first derivative IR fingerprint of Trichosanthes and their steamed products was calculated by using the Computer Aided Similarity Evaluation System. The principal component analysis(PCA)model and the partial least squares dis-criminant analysis(PLS-DA)model of IR spectral data of Trichosanthes before and after steaming were established by using SIMCA-P 11 statistical software for PCA and PLS-DA,and the classification of major compounds affecting information difference in IR spectra of Trichosanthes before and after steaming was selected by 3D scatter plot,load 3D scatter plot and variable important in project(VIP) value. Results The similarity between the original IR fingerprint and first derivative IR fingerprint of Trichosanthes and their steamed products were 0.9165 and 0.2832. Seven VIP>1 spectral peaks were screened out by using SIMCA-P 11 statistical software,of which,the absorption peak of 1456 cm-1 was νc=c,the absorption peak of 1726 cm-1 was νc=o and the VIP values were 1.6290 and 1.4256 respectively. Conclusion The categories of compounds of Trichosanthes before and after steaming did not change,but the chemical components changed. Compounds of Trichosanthes before and after steaming affect the difference in IR spectral information may mainly contain C=C-C=C or C=O or both of them.

Objective To analyze the changes in the chemical components in Fructus Trichosanthes before and after the pro?cessing of steaming,so as to explore the material basis of the pharmacodynamic changes between Trichosanthes and steamed Tricho-santhes.Methods The peaks matching data of Fructus Trichosanthes and steamed Fructus Trichosanthes were obtained by using the similarity evaluation system for chromatographic fingerprints of traditional Chinese materia medica.The principal component analysis (PCA)model and the partial least squares discriminant analysis(PLS-DA)model for the analysis of the Fructus Trichosanthes and steamed Fructus Trichosanthes data were established using the SIMCA-P 11 statistical software for PCA and PLS-DA,from which the score chart,load chart and Variable Importance(VIP)value were obtained,so as to identify the main different components in Fructus Trichosanthes and steamed Fructus Trichosanthes.Results The PCA(R2X=0.96,Q2=0.552)model and PLS-DA(R2Y=0.917,Q2=0.579)model were established,and 8 chromatographic peaks with significant difference in peak area were selected.Among them,two of the chromatographic peaks were assigned to be 5-hydroxy methyl furfural and vanilla acid,and 5-hydroxy methyl furfural had the largest VIP value.In addition,an unknown component was also found in the steamed Fructus Trichosanthes,which was generated in the process of steaming and needed to be identified in future studies.Conclusion The content of some chemical components in Fruc?tus Trichosanthes were changed after the process of steaming,and the processing of steaming also caused the formation of an unknown chemical component.5-Hydroxy methyl furfural and vanillic acid seem to be a likely choice for exploring the material basis of the phar?macodynamic changes in Fructus Trichosanthes after the processing of steaming.

Objective To investigate the sanitation status of centralized air ventilation system in Putuo District of Shanghai, to analyze correlation between different indexes and to provide basis for improving the health status of public places and strengthening health supervision. Methods According to the requirements of WS 394-2012 The Sanitation Criterion of the Centralized Air Ventilation System in Public Places, 18 centralized air ventilation systems in public places in Putuo District of Shanghai were randomly selected for sampling, testing and evaluation during 2014-2018, and 9 indexes of air supply, inner surface of air duct and indoor air were analyzed for correlation. Results The overall qualified rate of centralized air ventilation system in 10 hotels, 4 restaurants and 4 supermarkets was 86.0%.The over standard rate of PM₁₀, bacteria and fungi in air supply was 3.4%, 31.8% and 33.0%, respectively, but β-hemolytic streptococcus was not detected.The dust amount and microbial index on the inner surface of the air duct were up to standard.Another two indexes of fresh air quantity and Legionella pneumophila were tested in the hotel.The qualified rate of fresh air quantity was 25.6%, the positive rate of Legionella pneumophila type 1(LP1) in cooling water was 30.0%, and no Legionella pneumophila was detected in condensate water.The correlation analysis of 9 indexes showed that there was a significant positive correlation between PM₁₀ in air supply and in indoor air; the amount of microbial pollution on the inner surface of air duct and in air supply; and the amount of microbial pollution in air supply and in indoor air. Conclusions In Putuo District, the general sanitary condition of the centralized air ventilation system in public places is not to be optimistic.The qualified rate of bacteria and fungi in the air supply and fresh air quantity is low, and Legionella pneumophila was detected.The indoor air quality can be improved by installing efficient air purification device and regular cleaning and disinfection.

Objective The immunotoxins generated by conjugating monoclonal antibody (mAb) and a certain toxin play an important and promising role in treating hematopoietic malignancies. However, most of the toxins used for the conjugation are toxic proteins, which are immunogenic in the patients. Norcantharidin (NCTD) is a small molecule toxin without immunogenicity, and thus has become a potential new drug for hematopoietic cancers. In this study, we prepared immunotoxin 2E8-NCTD by using the ZCH-4-2E8 cells produced in the laboratory of our hospital, and then detected its targeting effect against CD+19 lymphoid malignant Nalm-6 cells in vitro.Methods 2E8 mAb was obtained from mouse ascites and purified by gel chromatography. After its purity was checked by SDS-PAGE, immunotoxin 2E8-NCTD was generated by conjugating CD19 mAb with NCTD using activated ester method. The binding activity of the immunoconjugate to CD19 antigens on cell surface, and the expression levels of the CD19 antigens on Nalm-6 and K562 cells were determined by flow cytometry. The inhibitory effects of PBS, purified 2E8 mAb, NCTD, and immunotoxin 2E8-NCTD on the cell growth of either Nalm-6 or K562 cells were then compared.Results The purity of the 2E8 mAb was higher than 99% demonstrated by SDS-PAGE assay. 2E8 mAb was detected on the surface of 99.34% of the Nalm-6 cells, while on only 0.98% of the K562. The newly generated immunotoxin had a positive rate of 99.90% on the Nalm-6 with slightly reduced binding activity. Both 2E8-NCTD and NCTD significantly inhibited the growth of CD+19 Nalm-6 cells (P < 0. 001 ), while the purified 2E8 mAb did not show any significant influences on the growth of the same cell line ( P > 0.05 ). Meanwhile, no significant inhibitory effects on the CD-19 K562 cells were identified in the 2E8-NCTD, 2E8 mAb, or control groups, indicating a significant targeting effect of 2E8-NCTD against Nalm-6 cells.Conclusions The immunotoxin 2E8-NCTD can be synthesized by activated ester method. It has target killing effects on CD+19 Nalm-6 leukemia cells in vitro.

OBJECTIVETo summarize the clinical presentation, pathogenesis, neuroimaging, treatment, and outcome of stroke-like migraine attacks after radiation therapy (SMART) syndrome, and to propose diagnostic criteria for this disorder.

DATA SOURCESWe searched the PubMed database for articles in English published from 1995 to 2015 using the terms of "stroke-like AND migraine AND radiation." Reference lists of the identified articles and reviews were used to retrieve additional articles.

STUDY SELECTIONData and articles related to late-onset effects of cerebral radiation were selected and reviewed.

RESULTSSMART is a rare condition that involves complex migraines with focal neurologic deficits following cranial irradiation for central nervous system malignancies. The recovery, which ranges from hours to days to weeks, can be partial or complete. We propose the following diagnostic criteria for SMART: (1) Remote history of therapeutic external beam cranial irradiation for malignancy; (2) prolonged, reversible clinical manifestations mostly years after irradiation, which may include migraine, seizures, hemiparesis, hemisensory deficits, visuospatial defect, aphasia, confusion and so on; (3) reversible, transient, unilateral cortical gadolinium enhancement correlative abnormal T2 and fluid-attenuated inversion recovery signal of the affected cerebral region; (4) eventual complete or partial recovery, the length of duration of recovery ranging from hours to days to weeks; (5) no evidence of residual or recurrent tumor; (6) not attributable to another disease. To date, no specific treatment has been identified for this syndrome.

CONCLUSIONSSMART is an extremely rare delayed complication of brain irradiation. However, improvements in cancer survival rates have resulted in a rise in its frequency. Hence, awareness and recognition of the syndrome is important to make a rapid diagnosis and avoid aggressive interventions such as brain biopsy and cerebral angiography.

Central Nervous System Neoplasms ; therapy ; Female ; Humans ; Male ; Migraine Disorders ; diagnosis ; etiology ; Radiation Injuries ; complications ; diagnosis ; Stroke ; diagnosis

OBJECTIVETo establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay.

METHODSRat kidney samples were hydrolyzed in 6 mol x L⁻¹ HCl under 110 degree for 24 h, then 9-fluorenylmethyl chloroformate (FMOC) was added for the pre-column reaction. The HPLC analysis was performed on a Phenomenex C₁₈ column using sodium acetate buffer, methanol and acetonitrile as mobile phase with a flow rate of 1.0 ml min⁻¹ and UV detective wavelength 265 nm at 40 degree.

RESULTSLinear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%.

CONCLUSIONThe established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.

Animals ; Chromatography, High Pressure Liquid ; methods ; Hydroxyproline ; analysis ; In Vitro Techniques ; Kidney ; chemistry ; Male ; Rats

OBJECTIVEThis review aims to illustrate the relationship between clinical features and the prognosis of patients with limb-shaking transient ischemic attack (LS-TIA).

DATA SOURCESRelevant articles published in two main Chinese medical periodical databases (China National Knowledge Infrastructure and China Science Periodical Database) from 1986 to June 2013 were identified with keywords "limb shaking" and "transient ischemic attack".

STUDY SELECTIONOriginal articles and case reports about LS-TIA were selected.

RESULTSA total of 63 cases collected from 19 articles were included in the pooled analysis. LS-TIA presented in two cerebrovascular diseases, of which atherosclerotic high-grade stenosis or occlusion in carotid artery system and moyamoya disease formed 95.2% and 4.8%, respectively. Of 63 patients, 11 (17.5%) were once misdiagnosed as epileptic and prescribed useless antiepilepsy drugs. The multivariable Logistic regression model showed a significant protective effect of patients with revascularization therapy on prognosis, compared with patients treated with drugs (odds ratio 0.20, 95% CI 0.05-0.74, P = 0.016).

CONCLUSIONSChronic carotid artery system hypoperfusion can induce limb(s) shaking, followed by high possibility of ischemic stroke in the same brain territorial. Revascularization of the responsible artery may work better than conservative drug-based therapy.

Aged ; Extremities ; physiopathology ; Female ; Humans ; Ischemic Attack, Transient ; pathology ; physiopathology ; Male ; Middle Aged ; Prognosis

OBJECTIVETo investigate the in vitro effects of fludarabine on apoptosis and gene expression profile in human multiple myeloma (MM) cells.

METHODSCell growth was measured by a MTT assay. Cells apoptosis was evaluated by flow cytometry. The activation of caspase cascade was determined by Western blot analysis. The expression profile of apoptosis-related genes in human MM cells was detected by cDNA expression array system.

RESULTSThe growth of RPMI8226 and KM3 cells was suppressed by fludarabine treatment in a dose and time-dependent manner. After treatment with fludarabine for 24 h, the IC(50) for RPMI8226 cells was 2.13 µg/ml, and 0.36 µg/ml for KM3 cells. Apoptotic cells of RPMI8226 and KM3 increased in a dose- dependent manner after exposure to fludarabine for 24 h. Western blot analysis showed the activation of caspase-3 and PARP in the MM cells treated with fludarabine. The cDNA expression array showed that multiple pathways were involved in the apoptosis induced by fludarabine. Among 97 apoptosis-related genes, 25 genes were differently expressed and 13 expressions were up-regulated in fludarabine treated group, involving in Bcl-2 pro-apoptotic gene, tumor necrosis factor (TNF) and its receptor superfamily gene, and caspase recruitment domain family, 12 genes expression down-regulated, including Bcl-2 antiapoptotic gene, TNF superfamily and its receptor related factor gene and apoptosis inhibitor protein family.

CONCLUSIONFludarabine can significantly inhibit MM cell growth and induce apoptosis in vitro. The multiple pathways may be involved for the apoptosis in MM cells.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Multiple Myeloma ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Transcriptome

A method was established for the simultaneous analysis of 25 trace elements and heavy metals in polysccharides from Liuwei Dihuang prescription, including Li, Be, B, Ti, Mg, Al, V, Cr, Mn, Co, Fe, Ni, Cu, Zn, Ga, As, Sr, Cd, Sn, Sb, Ba, Hg, Tl, Pb, Bi. The different rate of elemental extraction in Al, Fe, Mg, B, Ti, Mn, Zn, Sr, Ba was made in water and different concentration of alcohol. The samples, digested via microwave, calibrated by internal standard elements such as Ge and In, with bush branches and leaves as the controlled reference standard, were inlet into ICP-MS to analyze the contents of the 24 trace elements and heavy metals. The detection limits of the 24 elements were in the range of 0.007-2.225 µg · L(-1), while the RSD was below ≤ 4. 0%, with their recovery ranging from 84. 1% to 116%. Big different of the elemental extraction rates could be found by using different ethanol solutions. The method is simple, rapid and accurate, and can be used for the quality control of trace elements and heavy metals in Liuwei Dihuang polysccharides. With the aid of the obtained result, we may increase the extraction of necessary element while making an attempt at multi-element speciation in polysccharides from Liuwei Dihuang.
Chemical Fractionation ; Drugs, Chinese Herbal ; chemistry ; Mass Spectrometry ; methods ; Polysaccharides ; chemistry ; isolation & purification ; Trace Elements ; chemistry ; isolation & purification