Objective To construct eukaryotic expression plasmid pEGFP-N1/EPO and to detect its expression in vitro in order to provide experimental evidence for further study on the treatment of hypoxic-ischemic brain damage.Methods The total RNA was extracted from rat kidney.EPO cDNA fragment was obtained by RT-PCR amplication,inserted into pEGFP-N1 vector and identified by restriction endonuclease digestion and nucleotide sequencing.MSCs cells were transfected with the plasmid using Lipofectamine 2000.The expression of EPO protein was detected by immunohistochemistry technique and RT-PCR.Results Confirmed by restriction endonuclease digestion and sequencing,the recombinant plasmid contained the sequence of rat EPO gene was successfully constructed.After transfection with the plasmid,EPO protein could be expressed in MSCs cells.Conclusion The constructed eukaryotic expression vector pEGFP-N1/EPO can express rat EPO in vitro.

Objective To construct the prokaryotic expression system of reteplase fusion protein and research the effect of chaperones on its renaturation. Methods Inserted the reteplase gene into the prokaryotie expression vector PET32a and then expressed it by the induction of IPTG in E. coli BL21. Researched the effect of chaperones on the renaturation of fusion protein by adding different chaperones. Results The analysis of SDS-PAGE and Western blot indicated that reteplase fusion protein was expressed correctly. Chaperones DsbA,pKJE7,pTf16 had the conspicu-ous effect on the renaturation of fusion protein. The result of activity assay indicated that the refolded reteplase fu-sion protein had fibrinolytic activity. Conclusion Chaperones can promote renaturation of reteplase fusion protein.

This article was aimed to evaluate the α-glucosidase inhibitory activity of extracts from the root of Salvia miltiorrhiza Bunge and different processed products. The α-glucosidase inhibitory activity was screened with acarbose as positive control and by α-glucosidase inhibitory model in v itro . The results showed that the ex-tracts from the root of S. miltiorrhiza and different processed products had inhibitory activity. And the activity was higher than that of acarbose. In addition to the MeOH extract of S. miltiorrhiza carbon, the inhibitory activity of MeOH extracts from other processed products were higher than that of MeOH extract of S. miltiorrhiza. The in-hibitory activity of petroleum ether extracts of different processed products were close to S. miltiorrhiza. EtOAC extracts were lower than that of S. miltiorrhiza. The n-BuOH extracts were higher than that of S. miltiorrhiza. The inhibitory activity of extracts was positively correlated with concentrations, and it depended on the concentra-tion. It was concluded that the processed products of S. miltiorrhiza can strengthen α-glucosidase inhibitory activ-ity in different degrees.

OBJECTIVE:To establish a method for the detection of volatile constituents from the leaves and fruits of Piper ni-grum. METHODS:HS-SPME-GC-MS was used. The chromatographic conditions:column was HP-5 MS quartz elastic capillaries, carrier gas was high purity helium(99.999%),flow rate was 1.0 mL/min,the inlet temperature was 250 ℃,initial temperature of column was 50 ℃(temperature programmed),split injection with split ratio of 10:1. MS conditions:ionization mode was electron impact ion source,ionization energy was 80 eV,ion source temperature was 230 ℃,quadrupole temperature was 150 ℃,trans-mission line temperature was 280 ℃,electron multiplier voltage was 1588 V,mass scanning range was m/z 30-400. The spectra were retrieved using RTLPEST3. L and NIST08. L,and the relative contents of the volatile constituents were determined by area normalization method. RESULTS:There were 28 volatile constituents in the leaves and 15 in the fruits,respectively accounting for 67.13% and 36.85%. The major volatile constituents of leaves were β-caryophyllene (15.72%),limonene (9.39%),3-carene (9.32%),β-pinene(6.80%),α-terpine(4.98%),etc.,the main volatile constituents of fruits were 1,7,7-trimethyl-2-vinylbicyclo [2.2.1]hept-2-ene(10.45%),espatulenol(8.28%),caryophyllene oxide(4.81%),etc. 5 constituents were owned in both. CON-CLUSIONS:The study basically clears the main volatile constituents from the leaves and fruits of P. nigrum,and verifies existing obvious differences.

Objective:To study the effect of IL-15 eukaryotic expression plasmid on the humoral immune responses to HBV surface antigen protein vaccine.Methods:Had constructed two of IL-15 eukaryotic expression plasmides.One is the eukaryotic expression plasmid of the IL-15 whole sequence,the other is the chimeric eukaryotic expression plasmid of the IL-2 signal peptide and IL-15 mature peptide sequence(IL-2s-15).The bioactivities of the expression products of the two plasmids were identified by CTLL-2 proliferation assay.Results:After IL-15 plasmid co-immunized with HBsAg,the titer of anti-HBsIgG was much higher than that of control( P 0.05),however,the anti-HBsIgG2a/IgG1 ratio was higher than that of control.Conclusion:IL-15 expression plasmids effect differently on the immune responses induced by protein vaccine.

OBJECTIVE:To investigate the effect of the extracts of Polygonum lapathifolium on the activity of ?-glucosidase.METHODS:Soxhlet extraction was applied to extract overground part of P.lapathifolium,and the inhibition effect of extracts on ?-glucosidase was determined with 96-microplate-based method,and the inhibitory kinetic characteristics of methanol extract was investigated using Lineweaver-Burk method.RESULTS:The extracts from P.lapathifolium showed good inhibitory activity.The methanol extract had the highest inhibitory activity (IC50=2.13 ?g?mL-1)followed by ethyl acetate extract (IC50=3.826 ?g?mL-1),petroleum ether extract (IC50=230.86 ?g ? mL-1).And they were all higher than that of acarbose (IC50=1 103.01 ?g?mL-1) as positive control.Inhibitory kinetics test indicated that methanol extract was noncompetitive inhibitor.CONCLUSION:The extracts of P.lapathifolium have good inhibition effect on the activity of ?-glucosidase with a good prospect of development and utilization.

OBJECTIVEThe effect of waste batteries leaching on the seedling growth and volatile constituents in leaves of Schizonepeta tenuifolia was assayed.

METHODThe different concentrations of waste batteries leaching on the seedling growth were discussed. Volatile compounds were analyzed by solid-phase micro-extraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS).

RESULTThe results indicated that S. tenuifolia showed resistance to heavy metal polluting, but the high rate of waste batteries leaching had the inhibiting effect to seedlings growth. The waste batteries leaching cause the major volatile constituents in leaves of S. tenuifolia was changed greatly under waste batteries leaching solution stress.

CONCLUSIONHeavy metal leached by waste batteries had great effect on growth of S. tenuifolia, reducing its value for food and medical purposes.

Lamiaceae ; chemistry ; drug effects ; metabolism ; Metals, Heavy ; toxicity ; Plant Leaves ; chemistry ; drug effects ; Seedlings ; drug effects ; metabolism ; Stress, Physiological ; Volatile Organic Compounds ; analysis ; Waste Disposal, Fluid ; Water Pollutants, Chemical ; toxicity

This study was aimed to analyze the volatile constituents from flower, stem tip and seed of Cucurbita moschata Duch.(Miben). The volatiles were analyzed by head-space solid micro-extraction, coupled with GC-MS and Kovats indices for the first time . The results showed that 22 compounds were identified from the flower , 20 from the stem tip and 21 from the seed of the C. moschata (Miben). The total essential constituents from each part were 91 . 89%, 89 . 24% and 96 . 26%, respectively . A total of 10 compounds in the flower and stem tip were mutual. And 3 compounds in the flower, stem tip and seed were mutual. It was concluded that the β-bourbonene (17.57%) and heneicosane (11.90%) were the highest components of the total essential constituents of the flower of C. moschata (Miben). Decanal (28.77%) was the highest components of the stem tip and hexadecanoic acid ethyl ester (29.12%), 2,3-butanediol (16.90%) and linoleic acid ethyl ester (16.52%) were the highest compo-nents of seed of C. moschata (Miben).

Iraq is a multi-ethnic, multi-religious federal country located in southwest Asia and northeast of the Arabian Peninsula. Medical insurance is based on the primary health care model, supplemented by private medical care. Traditional medicine in Iraq is dominated by Islamic medicine. With the support of the Chinese government, the first Traditional Chinese Medicine (TCM) center was established in 2000. However, due to safety issues, the management of the center had problems such as insufficient staff, limited medical experience, and non-persistentsupport. At present, TCM has not been included in the Iraqi medical insurance system, which has affected the people’s medical choice and hindered the development of TCM. It is recommended to strengthen the construction of existing TCM centers, improve the accessibility of TCM education, and strengthen the cooperation and exchanges of traditional medicine between the two countries to promote the spread and development of TCM in Iraq.

OBJECTIVETo study chemical constituents in nutshell of Trapa acornis and in vitro inhibitory activity against alpha-glucosidase.

METHODEtOAC and n-butanol extractive fractions were separated by chromatography and their structures were identified by multiple spectroscopic techniques.

RESULTNine compounds were separated, they were 4,23,24-trimethylcholest-22-en-3-ol (1), stigmasterol (2), alpha-amyrin (3), (+)-nyasol (4), oleanolic acid (5), ursolic acid (6), hederagenin (7), 3,23-dihydroxy-12-ursen-28-oic acid (8) and beta-daucosterol (9). Total extracts from T. acornis nutshells, petroleum ether fractions, acetic ether fractions and normal butanol fractions showed inhibitory activity against alpha-glucosidase. Compound 5 (IC50 2.88 mg x L(-1)) and 6 (IC50 4.42 mg L(-1)) showed stronger inhibitory activity against alpha-glucosidase.

CONCLUSIONAll compounds except compound 2 were separated from the genus for the first time, and compound 1-9 were separated from this plant for the first time.

Enzyme Inhibitors ; chemistry ; Glycoside Hydrolase Inhibitors ; Lythraceae ; chemistry