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 investigate the effects of Corbrin Shugan capsule on dimethylnitrosamine (DMN)-induced hepatic fibrosis in rats.

METHODSHepatic fibrosis was induced by DMN in AD rats. The serum concentrations of III pro-collagen (III PC),laminin (LN) and tissue inhibitor of metalloproteinase-1(TIMP-1) were determined with ELISA. The concentration of albumin (ALB) in sera and the content of hydroxyproline (Hyp) in liver tissues were determined with chemical colorimetric and HPLC, respectively. The fibrosis area was measured with Motic Med 6.0 digital medical image analysis system.

RESULTSCompared to model group the high-dose (450 mg kg(-1)),mid-dose (270 mg kg(-1)) and low-dose (90 mg kg(-1)) groups of Corbrin Shugan capsule had significantly lower serum content of III PC [34.46 ± 13.95),(36.15 ± 9.46), and (40.58 ± 7.72)ng ml(-1) compared with (49.38 ± 10.95)ng ml(-1),P<0.05 or P<0.01],TIMP-1 [(16.65 ± 4.24),(16.66 ± 4.34),and (18.99 ± 6.05)ng ml(-1) compared with (30.84 ± 14.48)ng ml(-1), P<0.05 or P<0.01], LN [(12.94 ± 4.29), (12.96 ± 3.21),and (15.32 ± 8.00)ng ml(-1) compared with (30.22 ± 17.00)ng ml(-1),P<0.05 or P<0.01] and smaller hepatic fibrosis area [(0.02240 ± 0.01337), (0.02176 ± 0.01460) and (0.02384 ± 0.01405)μm(2) compared with vs (0.03929 ± 0.01732)μm2, P<0.05 or P<0.01]; the high-dose and mid-dose groups of Corbrin Shugan capsule had significantly lower content of Hyp in liver tissues [(0.77 ± 0.09) and (0.81 ± 0.09)μg μmg(-1) compared with (1.06 ± 0.33)μg mg(-1),P<0.05 or P<0.01]; and the high-dose group of Corbrin Shugan capsule significantly increased the content of ALB in sera [(34.02 ± 4.17)g L(-1) compared with (30.25 ± 4.21)g L(-1),P<0.05].

CONCLUSIONCorbrin Shugan capsule is effective in treatment of DMN-induced hepatic fibrosis in rats.

Albumins ; metabolism ; Animals ; Capsules ; Collagen Type III ; blood ; Dimethylnitrosamine ; adverse effects ; Drugs, Chinese Herbal ; therapeutic use ; Hydroxyproline ; metabolism ; Laminin ; blood ; Liver Cirrhosis, Experimental ; drug therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; blood

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

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

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

OBJECTIVETo observe the expression of GRP78 (glucose regulated protein, GRP78), Caspase-12 and the change of neuron apoptosis in the juvenile rat hippocampus after status convulsive (SC), and to explore the effect of edaravone on them.

METHODSOne hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into normal saline control group (NS group), status convulsive group (SC group) and edaravone treatment group (ED group). Each group was further divided into five subgroups in different executed time points after SC. The rats in status convulsive group were kindled into epilepsy by lithium-pilocarpine method. Expression of GRP78 mRNA and caspase-12 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) method. Expressions of GRP78 and caspase-12 protein were detected with immunohistochemical methods. The neuron apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL).

RESULTS(1) Measured by immunohistochemistry the value of OD of GRP78 (0.1480 ± 0.0164, 0.1682 ± 0.0114, and 0.1540 ± 0.0102, respectively, 12 h - 48 h points) and caspase-12 (0.1325 ± 0.0165, 0.1794 ± 0.0213, 0.1525 ± 0.0423, and 0.1309 ± 0.0199, respectively, 12 h-72 h points) positive cells in the SC group increased, there was a significant difference compared with NS group (GRP78: 0.1214 ± 0.0147, 0.1272 ± 0.0177, and 0.1260 ± 0.0157, respectively, 12 h-72 h points. Caspase-12: 0.1050 ± 0.0121, 0.1041 ± 0.0151, 0.1058 ± 0.0222, and 0.1036 ± 0.0186, respectively, 12 h - 72 h points) (P < 0.01, or P < 0.05). By ED intervention GRP78 (0.1550 ± 0.0131, 0.1886 ± 0.0154, and 0.1721 ± 0.0151, respectively, 12 h - 48 h points) positive cells value of the OD increased as compared with SC group (P < 0.01, or P < 0.05). and caspase-12 (0.1211 ± 0.0184, 0.1545 ± 0.0205, and 0.1085 ± 0.0219, respectively, 12 h, 24 h and 72 h points) positive cells value of the A decreased as compared with SC group (P < 0.01, or P < 0.05). (2) Measured by RT-PCR, the expression of GRP78 mRNA and caspase-12 mRNA trend was similar to protein. (3) The TUNEL positive cells in hippocampus CA(1) of SC group (11.41 ± 2.37) were more than that of NS group after the SC 12 h (P < 0.01), reached its highest level at 48 h (28.78 ± 5.11), after the intervention with edaravone (8.98 ± 2.22, 13.09 ± 2.54 and 20.57 ± 4.89, respectively, 12 h-48 h points), TUNEL positive cells showed a significant drop in SC group at 12 h-48 h time points (P < 0.01, or P < 0.05), but still significantly higher than that of the NS group (6.22 ± 1.50, 6.57 ± 1.61 and 6.72 ± 1.14, respectively) (P < 0.01, or P < 0.05), at the 4 h time point (NS group 6.29 ± 1.49, SC group 6.61 ± 1.71, ED group 5.75 ± 1.41) among the three groups, no significant difference in TUNEL positive cells was found (P = 0.759).

CONCLUSIONSThe expression of GRP78 and caspase-12 increased after SC. Edaravone increased expression of GRP78 and decreased expression of caspase-12 in hippocampus rat with pilocarpine-induced seizures, reduced the number of neuronal apoptosis. These results suggest that edaravone may have protective effect against the hippocampal damage caused by status convulsive.

Animals ; Antipyrine ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Caspase 12 ; metabolism ; Heat-Shock Proteins ; metabolism ; Hippocampus ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Seizures ; metabolism