Acupuncture Therapy ; Adult ; Aged ; Chronic Disease ; therapy ; Female ; Gastritis, Atrophic ; therapy ; Humans ; Male ; Middle Aged

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

OBJECTIVE:To establish the HPLC fingerprints for Sini decoction and compare the differences of compositions of Sini decoction prepared by traditional decoction and modern machine decoction. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.1% phosphoric acid(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 235 nm,column temperature was 30 ℃,and injection volume was 10 μl. The HPLC fingerprints of 10 batch-es of Sini decoction were determined with reference peak of liquiritin peaks,and common peak identification and similarity evalua-tion were conducted by using Chromatographic Fingerprint Similarity Evaluation System(2004 A edition). RESULTS:There were 18 common peaks and the similarity was no less than 0.982. According to the verification,the fingerprint of 10 batches of Sini de-coction showed good similarity with reference fingerprint,and the similarity of 10 batches of Sini decoction was high,which was prepared by the 2 methods. CONCLUSIONS:The established fingerprint is specific and stable,and can provide reference for quali-ty evaluation and control for Sini decoction;and there are no obvious differences in the main chemical compositions of Sini decoc-tion prepared by traditional decoction and modern machine decoction.

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

Background Researches determined that the alteration of A69S locus of age-related maculopathy susceptibility 2 ( ARMS2 ) gene is closely associated with the pathogenesis and progression of age-related maculopathy ( AM D ).However,the location of ARMS2 protein in normal eye tissue is still in controversy,therefore,its function is below understanding up to now.Objective The goal of this laboratory work was to investigate the distribution,expression and location of ARMS2 protein in normal adult retina and choroid as well as in retinal pigment epithelial (RPE) cells and lay a basis for exploring further its function in the protein level.Methods Ten donor eyeballs of normal adult male with the age from 28-42 years were collected in eye bank of Qingdao Eye Hospital.The frozen sections of the retina and choroid were prepared for the detection and location of ARMS2 in 3 eyes by immunofluorescence under the confocal laser microscope.The retina was isolated for the primary culture of RPE cells using explant culture method.The cells were then identified by CK32 antibody by immunofluorescence.The distribution and expression of the ARMS2 protein in retina,ehoroid and RPE cells were determined by immunofluorescence technique.Results ARMS2 protein was strongly expressed in retinal vessel,RPE cell layer,Bruch membrane and choroidal vessel,but weak expression was in retinal ganglion cell layer,inner nuclear layer,outer plexiform layer,outer nuclear layer and inner plexiform layer in the normal eyes.The primarily cultured cells appeared the polygon shape with the abundant pigment in cytoplasm.The immunofluorescence of the cells showed the positive response for CK32,exhibiting the green fluorescence granules in the cytoplasm.The positive expression of ARMS2 protein also was seen in the cytoplasm of RPE cells,appearing the red fluorescence.Conclusions ARMS2 protein mainly distribute and locate retinal and choroidal vessels,RPE cells and Bruch membrane in normal eye.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.

It has been a general trend to open the 5+3 medical personnel training mode of radiology in China.But at present,there are still a lot of problems during the process,such as,lack of training plan,unified teaching materials and examination index for reference,lack of learning consciousness and self-discipline among the students who participate in the standardization training,lack of adequate teaching resources in the standardization training base,and so on.In order to cultivate qualified and high-quality radiologists,this paper discussed the ways of forming a new set mode of 5+3 medical personnel training in the radiology department through exploration and practice,including strengthening teachers' construction and building a professional teachers troop,reforming the teaching method of standardization training,improving the teaching base infrastructure,establishing and perfecting the assessment evaluation mechanism.

Objective:To observe the effect of acupuncture on blood oxygen concentration in the brain of rats with post-traumatic stress disorder (PTSD) based on functional near-infrared spectroscopy (fNIRS),thus to reveal the mechanisms of acupuncture in intervening the brain function of PTSD rats.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except the blank group,rats in the other groups all received incarceration plus electric shock for 7 d to prepare the PTSD animal model.One hour before the stress model was established,rats in each group received the designated intervention:rats in the blank group and the model group did not receive any intervention;rats in the grasping group received grasping and fixation;rats in the paroxetine group received paroxetine hydrochloride solution by intragastric administration;and rats in the acupuncture group received acupuncture.Six-day treatment was a course,with 2 courses of treatment conducted for a total of 12 d.After the modeling,rats in each treatment group received intervention for 5 d,and the fNIRS system was used to collect and record the changes in the concentrations of oxygenated hemoglobin (HbO2),deoxygenated hemoglobin (d-Hb) and total hemoglobin (t-Hb) of the involved rat's brain regions,and also to assess the brain function.Results:Compared with the blank group,the concentration of HbO2 was significantly increased,the concentration of d-Hb was significantly decreased,and the concentration of t-Hb was significantly increased in the model group and the grasping group after the intervention,and the differences were statistically significant (all P＜0.01).Compared with the model group,the concentrations of HbO2,d-Hb and t-Hb in the grasping group did not change significantly (all P＞0.05).Compared with the grasping group,the concentration of HbO2 was significantly decreased,the concentration of d-Hb was significantly increased,and the concentration of t-Hb was significantly decreased in the paroxetine group and the acupuncture group,and the differences were statistically significant (all P＜0.05).There were no significant differences in the concentrations of HbO2,d-Hb and t-Hb between the paroxetine group and the acupuncture group (all P＞0.05).Conclusion:Acupuncture can regulate the blood oxygen concentration in the brain of PTSD model rats,which may be an important mechanism of acupuncture in intervening the brain function in PTSD rats.

Objective To investigate the effects of different stimulation of electroacupuncture on the protein expressions of SOCS1 and SOCS3 in rabbits with acute facial nerve injury; To determine the better stimulation. Methods New Zealand rabbits were treated with special hemostatic forceps for 5 min, and the length of the lesion was about 2.5 cm. The model of facial nerve injury was induced. The experiment was divided into blank group, sham-operation group, model group, and electoracupuncture weak-, medium-, and strong-stimulation group. The model group received no intervention after surgery. After treatment, the damaged facial nerve of each group was intercepted. The protein expressions of SOCS1 and SOCS3 mediated by negative feedback regulation of JAK-STAT were detected by ABC-ELISA. Results Compared with the blank group, the protein expressions of SOCS1 and SOCS3 in the model group increased (P<0.01). Compared with the model group, the protein expressions of SOCS1 and SOCS3 protein in electroacupuncture weak- stimulation group decreased (P<0.01). Conclusion Electroacupuncture can make SOCS1, SOCS3 protein expressions normal for acute facial nerve injury, and acupuncture treatment effect does not increase with the increase of stimulation.

PURPOSE: The objective of this study was to determine the phenotypic characterization of ligamentum flavum cells from patients with ossification of the ligamentum flavum (OLF). MATERIALS AND METHODS: Ligamentum flavum tissues were harvested from OLF and non-OLF patients during surgery. OLF and non-OLF cells were isolated from explant cultures. Cultured cells were analyzed using immunofluorescence staining and reverse transcription-polymerase chain reaction. RESULTS: OLF cells exhibited various appearances compared with the typical fibroblast-like morphology of non-OLF cells. Expressions of collagen type I and collagen type III were observed in OLF and non-OLF cells. OLF cells uniquely expressed osteocalcin, which is a marker for osteoblasts, and collagen type II which is a marker for chondrocytes, whereas they were negative in non-OLF cells. CONCLUSION: These findings indicate that OLF cells have phenotypic characterization of osteoblasts and chondrocytes which could play a role in the pathophysiology of OLF.
Adolescent ; Adult ; Aged ; Cells, Cultured ; Collagen Type I/genetics/metabolism ; Collagen Type II/genetics/metabolism ; Collagen Type VI/genetics/metabolism ; Female ; Humans ; Ligamentum Flavum/metabolism/*pathology ; Male ; Microscopy, Fluorescence ; Middle Aged ; Ossification, Heterotopic/metabolism/*pathology ; Osteocalcin/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult