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

Action Potentials ; Animals ; Female ; Male ; Nifedipine ; pharmacology ; Rabbits ; Ureter ; drug effects ; physiology ; Urination ; drug effects ; Vitamin K 3 ; pharmacology

Nerve growth factor(NGF) was firstly discovered as a member of neurotrophin family,and the research and development of NGF has been lasting more than fifty years since its discovery.To this end,a two-step and high –yield chromatographic method which consists of cation ion-exchange chromatography and reversed-phase chromatography was reported to isolate recombinant human beta nerve growth factor(? –NGF) secreted by constructed Chinese hamster ovary cells(CHO/dhfr-) from the culture media.Through the process of purification,the purity of protein which was determined by SDS-PAGE and RP-HPLC has reached to 95%,and the recovery of ? –NGF routed by RP-HPLC could be 70%.Furthermore,the biological activity of final purified protein evaluated by PC12 cells and dorsal root ganglia(DRG) exhibited the same performance as the standard protein of ? –NGF bought from Sigma,which indicated that there is no loss of biological activity through the isolation process.The conclusion suggested that an economical isolation method of recombinant human ? –NGF could be practiced on the industrial process of purification.

Pestalotiopsis photiniae is one of the predominant pathogens of strawberry root rot disease. Based on preliminary research, it was proved that crude toxins were main pathogenic substances of the pathogen. For further investigation and utilization of toxins produced by this fungus, conditions of producing toxins were analyzed with the leaf disk method in this experiment. The result showed that pH values, cultural time, vibration, and tested temperatures obviously affected the production of toxins, except for light treatment. The most suitable culture conditions for the toxin production were pH 6.2, 25?C, darkness and stillness, for 5 d～7 d. Besides, it was discovered that crude toxins could significantly inhibit seed germination and elongation growth of roots or shoots for maize, rye and mung bean.

BackgroundResearches found that the posterior capsular opacification (PCO) after lensextraction is associated with the elevation of the transforming growth factor-β(TGF-β).To seek the drug for inhibitingproliferation of lens epithelial cells (LECs) is crucial in the treatment and prevention of PCO.ObjectiveThisstudy was to investigate the preventing effects of decorin on the proliferation of LECs.MethodsRabbit LECs wascultured and passaged.The LECs in growth phase were incubated in 96 well plate at the density of 8×106/L.Decorinwith the concentrations 0.1,1.0,10.0 mg/L was added into the medium for 24,48 and 72 hours respectively.0.1％DMSO was used at the same way as positive control group,and the regular cultured cells worked as blank controlgroup.The inhibitory rates of different concentrations of decorin on the growth of LECs were detected by MTT at 24,48and 72 hours after addition of decorin.The percentage of LECs in different cell cycles in various groups was assayedusing flow cytometry.TGF-β level in medium suspension was detected using ELISA.The expression of TGF-β mRNA in LECs was checked by RT-PCR,and α-SMA expression in LECs was determined using immunochemistry.Results ELISA assay showed a statistical difference in the TGF-β levels of different groups (F=39.24,P=0.03 ).The TGF-β levels in 1.0,10.0 mg/L decorin groups were significantly decreased in comparison with blank control group (P＜0.01) and 0.1 mg/L decorin group (P＜0.05 ).The inhibitory rates of decorin in the concentrations of ≥ 1.0 mg/L on the growth of LECs were higher than the blank control group,and those in various concentrations of decorin groups were considerably lower in 24 hours compared with 48 and 72 hours ( P＜0.05 ) and so was the 48 hours compared with 72 hours (P＜0.05 ).The percentages of LECs in G0/G1 phase were ascent in 0.1,1.0 and 10.0 mg/L decorin groups in comparison with G2/M and S phase (P＜0.05).Immunochemistry revealed the weak expression of α-SMA in various decorin groups in comparison with control group. Conclusions Decorin can effectively inhibit LECs growth and induce LECs apoptosis in concentration- and time-dependent manner.It is suggested that decorin can be used in the prevention and treatment of after cataract.

On account of the dense cuticles of the fresh stem and the light, hard and pliable texture of the dried stem, Dendrobii Caulis is difficult to dry or pulverize. So, it is very important to the ancient doctors that Dendrobii Caulis should be properly treated and applied to keep or evoke its medicinal effects. The current textual research results about the preliminary processing, processing and usage methods of Dendrobii Caulis showed that: (1) In history the clinical use of fresh or processed Dendrobii Caulis as teas and tinctures were very common. (2) Its roots and rhizomes would be removed before using. (3) Some ancillary approaches were applied to shorten drying times, such as rinsing with boiling mulberry-ash soup, washing or soaking with liquor, mixing with rice pulp and then basking, etc. (4) According to the ancients knowledge, the sufficient pulverization, by means of slicing, rasping, hitting or pestling techniques, was necessary for Dendrobii Caulis to take its effects. (5) The heat processing methods for Dendrobii Caulis included stir-baking, stir-frying, steaming, decocting and stewing techniques, usually with liquor as an auxiliary material. Among above mentioned, steaming by pretreating with liquor was most commonly used, and this scheme was colorfully drawn in Bu Yi Lei Gong Pao Zhi Bian Lan (Ming Dynasty, 1591 CE) ; moreover, decocting in advance or long-time simmering so as to prepare paste products were recommended in the Qing Dynasty. (6) Some different processing programs involving stir-baking with grit, air-tightly baking with ondol (Kangs), fumigating with sulfur, which appeared in modern times and brought attractive outward appearance of the drug, went against ancients original intentions of ensuring drug efficacy.
Dendrobium ; History, Ancient ; Medicine, Chinese Traditional ; history ; Technology, Pharmaceutical ; history

Endometriosis (EM) is one of the common and frequently encountered gynecological diseases that seriously influences women's health. Its morbidity reaches 10%-15% in women at reproductive ages, and shows an evident rising tendency. In recent years, the Chinese medicine treatment of EM has won favorable therapeutic effects with few adverse reactions. A brief review on this topic has been made through analyzing and summarizing recent pertinent literatures in terms of treatment depending on syndrome differentiation, cycle treatment, external treatment, integrative medicinal treatment, so as to try to know the status quo of Chinese medicine treatment on EM, and to provide some instructive views for clinical treatment and research.
Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endometriosis ; drug therapy ; etiology ; physiopathology ; Female ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; trends ; Menstruation ; drug effects ; physiology

BACKGROUNDFolic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.

METHODSMorpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.

RESULTSDHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.

CONCLUSIONSDHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; physiology ; Female ; Heart ; embryology ; Heart Defects, Congenital ; etiology ; Tetrahydrofolate Dehydrogenase ; genetics ; physiology ; Zebrafish ; Zebrafish Proteins ; genetics ; physiology

BACKGROUNDTbx1 is the major candidate gene for DiGeorge syndrome (DGS). Similar to defects observed in DGS patients, the structures disrupted in Tbx1(-/-) animal models are derived from the neural crest cells during development. Although the morphological phenotypes of some Tbx1 knock-down animal models have been well described, analysis of the cardiac performance is limited. Therefore, myocardial performance was explored in Tbx1 morpholino injected zebrafish embryos.

METHODSTo elucidate these issues, Tbx1 specific morpholino was used to reduce the function of Tbx1 in zebrafish. The differentiation of the myocardial cells was observed using whole mount in situ hybridization. Heart rates were observed and recorded under the microscope from 24 to 72 hours post fertilization (hpf). The cardiac performance was analyzed by measuring ventricular shortening fraction and atrial shortening fraction.

RESULTSTbx1 morpholino injected embryos were characterized by defects in the pharyngeal arches, otic vesicle, aortic arches and thymus. In addition, Tbx1 knock down reduced the amount of pharyngeal neural crest cells in zebrafish. Abnormal cardiac morphology was visible in nearly 20% of the Tbx1 morpholino injected embryos. The hearts in these embryos did not loop or loop incompletely. Importantly, cardiac performance and heart rate were reduced in Tbx1 morpholino injected embryos.

CONCLUSIONSTbx1 might play an essential role in the development of pharyngeal neural crest cells in zebrafish. Cardiac performance is impaired by Tbx1 knock down in zebrafish.

Animals ; Branchial Region ; cytology ; drug effects ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; In Situ Hybridization ; Myocardium ; cytology ; Neural Crest ; cytology ; drug effects ; Oligonucleotides, Antisense ; pharmacology ; T-Box Domain Proteins ; antagonists & inhibitors ; metabolism ; Thymus Gland ; cytology ; drug effects ; Zebrafish ; embryology ; metabolism ; Zebrafish Proteins ; antagonists & inhibitors ; metabolism

OBJECTIVETo examine the effect of particulate matter (PM) less than 10 microns in diameter (PM10) and ozone (O3) on daily mortality in Shanghai, China.

METHODSA generalized additive model with penalized spline function was used to observe the acute effect of PM10 and O3 on daily mortality.

RESULTSHigher PM10 significantly increased the effect of O3 on total mortality, and O3 also increased the effect of PM10 although the estimated increment was statistically insignificant.

CONCLUSIONOur findings provide further evidence for the effect of PM10 and O3 on daily mortality.

Air Pollutants ; toxicity ; China ; epidemiology ; Humans ; Mortality ; Ozone ; toxicity ; Urban Population