Objective To investigate the regulatory effect of miR-21 on ADAMTS-1 expression in pulmonary fibrosis model. Methods A mouse model of pulmonary fibrosis was established using bleomycin (BLM) anda model of pulmonary fibrosis was constructed in vitro , the expression level of miR-21 was measured by quantitative real-time polymerase chain reaction (qRT-PCR), while the protein expression of ADAMTS-1 was measured by Western blot. NIH3T3 were transfected with miR-21 mimics and inhibitor in vitro and the cellular expression of ADAMTS-1 was measured by Western blot. Results Compared with the blank group , in mouse models of pulmonary fibrosis , the miR-21 expressions in lung tissues at three time points after BLM-treatment were significantly up-regulated while an evident decrease in ADAMTS-1 expressions were observed (P < 0.01). In vitro pulmonary fibrosis model , NIH3T3 cells after TGF-β1 in concentration 5 μg/L stimulation down-regulated ADAMTS-1 expression and up-regulated miR-21 expression (P < 0.01). NIH3T3 transfected with miR-21 mimics and inhibitor, up-regulated miR-21 expression, while down-regulated ADAMTS-1 protein expression. Conclusions Up-regulation of miR-21 and Down-regulation of ADAMTS-1 might be involved in the progression of pulmonary fibrosis model; miR-21 could negatively regulate ADAMTS-1 expression.

Objective To evaluate the diagnostic value of MSCT in Solitary Pulmonary Nodules (SPN) with different pathologic types .Methods Four-five SPN with pathological findings including 16 benign cases and 29 malignant cases were retrospectively an-alyzed ,and performed with MSCT scanning .Observation of the distribution ,morphology and internal characteristics of SPN by post-processing technique were done ,and the results were compared with pathological findings .Results Among 45 SPN ,the benign nodules included 6 cases of inflammatory pseudotumor ,4 cases of tuberculoma ,3 cases of hamartoma ,2 cases of cryptococcus and 1 case of hemangioendothelioma ,and the malignant nodules included 21 cases of adenocarcinoma ,3 cases of squamous carcinoma ,2 ca-ses of small cell carcinoma and 3 cases of alveolar cell carcinoma .Average diameter of benign nodules was 2 .1 cm ,and 2 .6 cm for the malignant nodules .All SPN ,including 5 cases in the inner zone ,12 in the medial zone and 28 in the lateral zone ,the percentages with malignant signs at the corresponding parts were 40% (2/5) ,50% (6/12) ,75% (21/28) ,there was significant difference be-tween distributions(P<0 .05) .The distributions of malignant nodules in the upper and lower lung lobe had no significant difference (P>0 .05);the incidences of leaf sign ,burr sign ,pleural indentation sign ,cavitas and bronchial air sign ,blood vessels cluster sign of malignant nodules were higher than that of the benign nodules(P<0 .05) ,and the inflammatory pseudotumor and adenocarcinoma owned similar characteristics ;the incidence of calcification in tuberculoma and hamartoma was high ,while cavitas in tuberculosis and lung cancer were high .Conclusion MSCT can sufficiently display the distribution ,morphology and internal characteristics of SPN , and can help make qualitative diagnosis of benign and malignant SPN ,providing great practical value in clinical treatment .

Ten compounds were isolated from the artificially induced dragon's blood of Dracaena cambodiana by various column chromatographies on silica and sephadex LH-20 gel. Based on spectral analysis of NMR and MS, their structures were identified as 3, 4-dihydroxyallylbenzene (1), 3', 4', 5'-trimethoxycinnamylalcohol (2), pinoresinol (3), (2R)-7, 4'-dihydroxy-8-methylflavane (4), (2R)-7,4'-dihydroxy-5-methoxy-8-methylflavane(5),(2S)-7,3'-dihydroxy-4'-methoxy-8-methylflavane(6) ,(2S)-4',7-dihydroxy-6, 8-dimethylflavane(7), 4,2',4'-trihydroxychalcone(8), 4,4'-dihydroxy-2-methoxydihydrochalcon(9) and Cambodianin E (10). Antibacterial activity assay showed that compounds 1, 4 and 10 have inhibitory effect on Fusarium oxysporum f. sp. cuben, Fusarium oxysporum f. sp. niveum, Fusarium oxysporum f. sp. vasinfectum and Ralstonia solanacearum.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Dracaena ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fusarium ; drug effects ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Spectrometry, Mass, Electrospray Ionization

Objective To investigate the influences of survivin down-regulation on cell G2/M phase arrest,apeptosis and sensitivity to carbon ion irradiation. Methods Small interfering RNA (siRNA) targeting survivin mRNA was designed, in vitro chemo-synthesized and transfected into SMMC-7721 cells. Survivin mRNA expression in SMMC-7721 cells was measured by real-time PCR, and the apeptotic rates by Annexin-FTTC at 24 and 48 h after transfection. Cell G2/M phase arrest after transfection was assessed with flow eytometry as well. Cellular sensitivity to high-LET carbon ions was determined by means of colony-forming assay. Results The expressions of survivin at mRNA level were down-regulated to be 59% and 39% in relation to the non-treated cells at 24 and 48 h after siRNA transfeetion, respectively. G2/M phase arrest in SMMC-7721 cells at 24 h after transfection was observed while much more obvious at 48 h. The apeptotic rate of SMMC-7721 cells was 21.41 % at48 h after survivin siRNA transfection, which was significantly higher than that of the cells transfected with negative siRNA. Moreover, a decreased clonogenic survival in siRNA treated group was shown. Conclusion Down-regulation of survivin gene expression in SMMC-7721 cells by siRNA could effectively induce cell apeptosis and G2/M phase arrest, and enhance the cellular radiosensitivity to high-LET heavy ions.

HuaFu Shengji is the primary traditional Chinese medicine (TCM) therapy for treating chronic skin ulcer. The high activities of the protein enzyme in the wound fluids is one of the main cause of healing delay. In order to investigate the effect of TCM Zhuhong ointment for promoting wound healing. This research focused on its influence on matrix metalloproteinase (MMP) activities in wound fluids with TCM Yang syndromes, directly on the activated MMP-1,2 activities in vitro and on MMP-1,-2,-9 production by HSF. 8 wound fluid samples were collected, which were diagnosed Yang Syndromes in TCM. Wound fluid activities of MMP-2 and MMP-9 were measured by gelatin zymogram assay. MMP-1 and MMP-2 activities in vitro were measured by substrate cleavage. CCK-8 was used to observe the toxicity of Zhuhong ointment on HSF. MMP-1,-2,-9 production by HSF were detected by confocal microscope. Zhuhong ointment from 1 to 25 g x L(-1) obviously inhibited MMP-2 activity in wound fluid. When Zhuhong ointment was over 5 g x L(-1), it showed significantly inhibitory effect on wound fluid MMP-9 activity. In vitro study, when the mercury concentration was 320 mg x L(-1), Zhuhong ointment solution directly inhibited both MMP-1 activity and MMP-2. But mercury concentration from 0.51-2.56 mg x L(-1), it could activate MMP-1 activity, and from 0.51-64 mg x L(-1), activate MMP-2 activity instead. The mercury concentration when Zhuhong ointment saturated in DMEM was 39.6 mg x L(-1). When the mercury concentration was over 1.23 mg x L(-1), Zhuhong ointment showed toxicity to HSF. At 1.23, 0.62, 0.31 mg x L(-1) of mercury concentration, it increased MMP-1 expression by HSF, and at 1.23, 0.62 mg x L(-1), decreased MMP-2 expression. However, at 1.23, 0.62, 0.31 mg x L(-1), it decreased MMP-9 expression. At higher concentration, Zhuhong ointment can inhibit MMP-2, MMP-9 activities in wound fluid with dose-dependent way and show a direct inhibitory effect on activated MMP-1 and MMP-2 in vitro. But at a lower concentration, it showed two-way adjustment, with increased MMP-1, MMP-2 activities and its expression by HSF and decreased MMP-9 activity.
Body Fluids ; enzymology ; Cells, Cultured ; Dermatitis ; drug therapy ; enzymology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; drug effects ; enzymology ; physiology ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Wound Healing ; drug effects

White spot syndrome virus (WSSV) is a major pathogen in aquaculture penaeid shrimp, which caused catastrophic economic losses in the worldwide. No adequate treatments against WSSV are available. In order to study infection mechanism of WSSV, a phage display scFv cDNA library against WSSV was constructed and a neutralizing antibody of scFv P1D3 was selected in our lab previously. In this study, scFv P1D3 was expressed successfully in yeast. Firstly, the original expression vector of P1D3, M13 phagmid, was used as a template to design primers with restriction sites of SnaB I and EcoR I . Then the gene of P1D3 was amplified by PCR. After digested by SnaB I and EcoR I , the fragment of scFv P1D3 with E-tag was inserted into yeast and E. coli shuttle plasmid pPIC9k. The recombinant plasmid pPIC9k-scFv P1D3-Etag was linearized with Bgl II and then transformed into Pichia pastoris GS115 by electroporation. Positive clones were selected and verified by PCR and DNA sequencing. The scFv PID3 was induced to express in yeast by methanol. The results of ELISA demonstrate that scFv P1D3 expressed in yeast still has high specificity to bind on WSSV and the binding activity is higher than that expressed in E. coli TG1. After several optimizing experiments, the results show that the expression amount of scFv P1D3 can reach to 302 mg/L in yeast culture supernatant. This experiment has offered a new source of antibody for the researches on passive immunology for shrimp.
Animals ; Antibody Specificity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Methanol ; pharmacology ; Penaeidae ; virology ; Pichia ; drug effects ; genetics ; Single-Chain Antibodies ; analysis ; biosynthesis ; genetics ; immunology ; Temperature ; White spot syndrome virus 1 ; immunology

Using the absorbent resin, silica gel and ODS column chromatography as well as semi-preparative HPLC, ten compounds were isolated from 70% ethanol extract of tubers of Dioscorea zingiberensis C. H. Wright, and their structures were elucidated as trigoneoside XIIIa (1), parvifloside (2), trigoneoside IVa (3), deltoside (4), protobioside (5), lilioglycoside k (6), zingiberensis newsaponin I (7), deltonin (8), prosapogenin A of dioscin (9), and trillin (10) on the basis of NMR and MS spectral data analysis. Among these compounds, 1, 3, 5 and 6 were isolated from this plant for the first time. In the screening test on platelet aggregation, compounds 7 and 8 exhibited induction effect on platelet aggregation, while compound 9 exhibited significant inhibitory effect on platelet aggregation in vitro.
Animals ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rats ; Rats, Wistar ; Saponins ; chemistry ; pharmacology

OBJECTIVETo evaluate the value of ultrasonic monitoring in induced abortion during the first trimester.

METHODSTotally 110 healthy women with a singleton pregnancy between 9 and 11 gestational weeks were enrolled. All the procedures of induced abortion were performed routinely. Ultrasonography was performed when the procedure of induced abortion was completed. Patients with normal ultrasonographic results were assigned in the control group, while patients with abnormal ultrasonographic findings were enrolled in the study group,in which these patients underwent further operations until the ultrasonography showed clear endomembrane line. All the recurretaged tissue in the study group were sent for pathological examinations.

RESULTSOf these 110 patients, 28 (25.5%) entered study group and 82 (74.5%) entered control group. In the study group, trophoblastic cell or chorion was found in the recurretaged tissue in 11 patients (39.3%), in which 3 had trophoblastic cell embedded in smooth muscular tissue, 6 had pathologic deciduas, and 11 had pathologic secretory endometria. In this control group,1 patient (0.9%) had retained products of conception. The operation duration [(20.6∓2.1) min vs.(11.5∓3.5) min, P0.05].

CONCLUSIONSUltrasonic monitoring in induced abortion during the first trimester can decrease the incidence of retained products of conception and will not induce endometrial damage. It is especially useful for women whose fetuses were at older gestational ages.

Abortion, Induced ; methods ; Adult ; Female ; Humans ; Monitoring, Intraoperative ; methods ; Pregnancy ; Pregnancy Trimester, First ; Ultrasonography

ObjectiveTo investigate the development trend of drinking water type of endemic fluorosis in Qinghai province, and to provide the basis for the prevention and treatment of the disease. MethodsIn 2009, six monitoring counties were chosen by using simple random sampling methods, all diseased villages of the six monitoring counties were classified into light, moderate and severe disease types according to water fluorine content on the historical data, and 1 village was respectively chosen from each type. In monitoring villages with improved water, 3 tap water and one source water samples were collected, respectively. Five water samples were collected randomly in water unimproved monitoring villages according to water well locations of east, west, south, north and center. The fluorine content in water and urine was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006). Children aged 8 to 12 were examined for dental fluorosis by Dean method.Clinical osteofluorosis of all the resident over the age of 16 was examined, 2 village of these counties were randomly selected, and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray using Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008). Urine sample of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method (WS/T 89-2006). ResultsImproving water projects had been implemented in 14 monitoring villages of the 18 villages in 6 counties, the rate of improved-water was 77.78％(14/18). Among the 14 projects, 5 improved-water projects ran normally, and 9 projects ran with intermittently water supply. Seventy-five water samples were tested, themean of water fluoride was 0.48 mg/L. The prevalence of dental fluorosis was 31.95％ (285/892), that of clinical skeletal fluorosis was 36.55％(1570/4295) and the X-ray detection rate of skeletal fluorosis was 25.64％ (20/78).Five hundred and seventy-one urine samples of children were determined, and geometric mean of urinary fluorine was 1.04 mg/L; 370 adult urine samples were determined, and geometric mean of urinary fluorine was 1.52 mg/L Conclusion Epidemic of drinking water type of endemic fluorosis is still serious in Qinghai province, and drinking water defluoride measures should be further strengthened and improved.

Objective To investigate the prevalence change of drinking water type of endemic fluorosis and the effect of control measures implemented in Huangyuan county of Qinghai province. Methods In 2009, all the endemic fluorosis villages in Huangyuan county were divided into two degrees, light and medium, according to the water fluorosis content before implementing the improving water project, 1 to 2 villages were selected from each degree village, respectively,as monitoring sites, and a total of 3 villages were selected. Source water and tap water samples were collected from each village and water fluoride concentration was determined. Dental fluorosis of all children aged 8 to 12 of monitoring villages was examined, and urine samples were collected by age group of children for determination of urinary fluoride. Clinical skeletal fluorosis of adults over 16 years of age was examined, and 20 copies of adults urine samples were collected to determine urinary fluoride. One village was selected in the 3 villages monitored to conduct X-rays examination of skeletal fluorosis. Water fluoride was tested in accordance with the "Non-metallic Targets Test Methods for Drinking Water" (GB/T 5750.6-2006); urinary fluoride was tested by fluoride ion-selective electrode method (WS/T 89-1996); dental fluorosis was diagnosed using Dean method;adult skeletal fluorosis was diagnosed by "Clinical Diagnostic Criteria for Endemic Skeletal Fluorosis"(WS 192-2008). Results Twelve water samples were assayed, water fluoride was (0.35 ± 0.43) mg/L. The detectable rate of dental fluorosis of 122 children aged 8-12 was 34.43%(42/122) and the geometric mean urinary fluoride was 0.89 mg/L of the 96 children. Of the 834 adults aged 16 and over, clinical detection of skeletal fluorosis was 47.72% (398/836) and geometric mean urinary fluoride was 1.10 mg/L of the 65 cases of adult urine samples assayed, detection rate of X-rays was 31.4% (11/35) in Gangou village of the 35 adults examined.Conclusions In Huangyuan county, water fluoride of the 3 surveyed villages are normal but the endemic fluorosis is still serious. It should strengthen monitoring and analyze the causes and improve prevention measures.