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.

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

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

Objective To investigate proton magnetic resonance spectroscopy(~1H-MRS)findings and value on dog's brain contusion and laceration.Methods Models of focal brain contusion and laceration in 10 dogs were established through hitting on the right frontal-parietal lobe with a freely drop of 200g weight at 1.3 m height.Serial examinations(1 h,24 h,72 h,5 day,8 day and 14 day after trauma)were performed with conventional MRI and ~1H-MRS.NAA/Cr,Cho/Cr and NAA/Cho rates were analyzed with GE system 1.5 T scanner and relative software.After examination,all dogs were executed to death. Pathological study was performed at local brain contusion.Results 1 h and 24 h-post trauma,NAA/Cr、Cho/Cr、NAA/Cho were significantly reduced(NAA/Cr 0.843?0.214,0.862?0.204,contralateral ones 1.069?0.284,1.048?0.232,t=-7.227,-6.718,Cho/Cr 1.181?0.224,1.243?0.134,contralateral 1.415?0.305,1.455?0.159,t=-4.332,-4.489,NAA/Cho 0.701?0.147,0.536?0.136, contralateral 0.832?0.245,0.613?0.165,t=-2.652,-2.665.P＜0.05).Microscopy showed focal petechial hemorrhage and necrosis,neuron loss,neuraxonal swelling and small glial cell slightly hyperplasia.Five day post trauma,Cho/Cr was significantly elevated in comparison with contralateral ones (1.517?0.197,contralateral 1.387?0.214,t=3.758.P＜0.05).Pathologically,inflammatory was obvious,peri-angiitis,granula tissue and fibrosis were seen.8—14 day later,NAA/Cr was not significantly reduced(0.895?0.105,0.875?0.153,contralateral 0.989?0.169,0.990?0.173,t=-2.909, -2.471.P＞0.05),Cho/Cr was significantly increased(1.457?0.168,1.572?0.374,contralaterl 1.334?0.174,1.366?0.352,t=7.312,3.201.P＜0.05)Inflammatory and gila1 hyperplasia was more significant,granuloma were seen.Lipid and Lac peak were not seen at all stages.Conclusion MRS could be a methods to monitor neuron injury and repair,and dynamically to detect the metabolic changes of brain contusion and laceration,reflecting injury severity and provide theory data for early treatment and predicting long-term outcome after trauma.

OBJECTIVETo study the expression levels of monokine induced by interferon-gama; (Mig) mRNA and its association with HBV DNA and alanine aminotransferase (ALT) in patients with chronic hepatitis B.

METHODSThe level of Mig mRNA in peripheral blood mononuclear cells (PBMCs) was dynamically detected with real-time quantitative PCR, and the ratio of chemokine/GAPDH was considered to represent the final chemokine level. The plasma level of was measured by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe mean level of Mig mRNA in PBMCs of the patients with chronic hepatitis B was 0.6883-/+0.0693, which was significantly higher than that in normal controls (P<0.001). The plasma Mig level in the patients was 609.6-/+73.8 pg/ml, also significantly higher than that in normal controls (P<0.05). In patients with chronic hepatitis B, the level of Mig mRNA in the PBMCs was significantly correlated with plasma Mig level (r=0.7157, P<0.001), and plasma Mig level was correlated with plasma ALT level (r=0.7220, P<0.001) and plasma HBV DNA level (r=0.7266, P<0.001).

CONCLUSIONBoth the expression of Mig mRNA in PBMCs and plasma Mig concentration are elevated in patients with chronic hepatitis B. Mig plays an important role in migration of the inflammatory cells to the liver and mediates the development of chronic hepatitis B.

Adolescent ; Adult ; Chemokine CXCL9 ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B, Chronic ; blood ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the effect of methotrexate (MTX), a folic acid antagonist which can lead to folic acid deficient, on the cardiac development and on the expressions of BMP2b and HAS2 in zebrafish.

METHODSThe zebrafish embryos at 6-48 hrs post fertilization (hpf) were treated with various concentrations of MTX (0.5 x 10(-3), 1.0 x 10(-3) and 2.0 x 10(-3) M). At 48 hpf, the percentage of cardiac malformation and heart rate were recorded. The zebrafish embryos at 6-10 hpf treated with 1.5 x 10(-3) M MTX were used as the MTX treatment group. At 24 and 48 hpf the cardiac morphology was observed under a microscope. The expressions of BMP2b and HAS2 in zebrafish were detected by in situ antisense RNA hybridization and real-time PCR.

RESULTS6-12 hpf, the early embryonic developmental stage, was a sensitive period that MTX affected cardiac formation of zebrafish. The retardant cardiac development and the evidently abnormal cardiac morphology was found in the MTX treatment group. The results of in situ antisense RNA hybridization showed that the expressions of BMP2b and HAS2 in the zebrafish heart were reduced in the MTX treatment group at 36 and 48 hpf. The real-time PCR results demonstrated that the BMP2b expression decreased at 12, 24, 36 and 48 hpf, and that the HAS2 expression decreased at 24, 36 and 48 hpf in the treatment group compared with the control group without MTX treatment.

CONCLUSIONSThe inhibition of folic acid function may affect cardiac development of early embryos, resulting in a retardant development and a morphological abnormality of the heart in zebrafish, possibly by down-regulating the expressions of BMP2b and HAS2.

Abnormalities, Drug-Induced ; etiology ; Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Down-Regulation ; Folic Acid Antagonists ; toxicity ; Gene Expression Regulation ; drug effects ; Glucuronosyltransferase ; genetics ; Heart Defects, Congenital ; chemically induced ; Hyaluronan Synthases ; Methotrexate ; toxicity ; Polymerase Chain Reaction ; Zebrafish ; Zebrafish Proteins ; genetics