For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Catalytic activity of activated carbon supported tungstosilicic acidin synthesizing 2-methyl-2-ethoxycarbonylmethyl1,3-dioxolane, 2,4-dimethyl-2-ethoxycarbonylmethyl-l,3-dioxolane, cyclohexanone ethylene ketal, cyclohexanone 1,2-propanediol ketal, butanone ethylene ketal, butanone 1,2-propanediol ketal, 2-phenyl-1,3-dioxolane, 4-methyl-2-phenyl-1,3-dioxolane,2-propyl-1,3-dioxolane, 4-methyl-2-propyl-1,3-dioxolane was reported. It has been demonstrated that activated carbon supported tungstosilicic acid is an excellent catalyst. Various factors involved in these reactions were investigated. The optimum conditions found were: molar ratio of aldehyde/ketone to glycol is 1/1.5, mass ratio of the catalyst used to the reactants is 1.0%, and reaction time is 1.0 h. Under these conditions, the yield of 2-methyl-2-ethoxycarbonylmethyl-l,3-dioxolane is 61.5%, of 2,4-dimethyl2-ethoxycarbonylmethyl-1,3-dioxolane is 69.1%, of cyclohexanone ethylene ketal is 74.6%, of cyclohexanone 1,2-propanediol ketal is 80.1%, of butanone ethylene ketal is 69.5%, of butanone 1,2-propanediol ketal is 78.5%, of 2-phenyl-1,3-dioxolane is 56.7%, of 4-methyl-2-phenyl- 1,3-dioxolane is 86.2%, of 2-propyl-1,3-dioxolane is 87.5%, of 4-methyl-2-propyl-1,3-dioxolane is 87.9%.

Catalytic activity of activated carbon supported tungstosilicic acid in synthesizing 2-methyl-2-ethoxycarbonylmethyl- 1,3-dioxolane, 2,4-dimethyl-2-ethoxycarbonylmethyl-1,3-dioxolane, cyclohexanone ethylene ketal, cyclohexanone 1,2-propa- nediol ketal, butanone ethylene ketal, butanone 1,2-propanediol ketal, 2-phenyl-1,3-dioxolane, 4-methyl-2-phenyl-1,3-dioxolane, 2-propyl-1,3-dioxolane, 4-methyl-2-propyl-1,3-dioxolane was reported. It has been demonstrated that activated carbon supported tungstosilicic acid is an excellent catalyst. Various factors involved in these reactions were investigated. The optimum conditions found were: molar ratio of aldehyde/ketone to glycol is 1/1.5, mass ratio of the catalyst used to the reactants is 1.0%, and reaction time is 1.0 h. Under these conditions, the yield of 2-methyl-2-ethoxycarbonylmethyl-1,3-dioxolane is 61.5%, of 2,4-dimethyl- 2-ethoxycarbonylmethyl-1,3-dioxolane is 69.1%, of cyclohexanone ethylene ketal is 74.6%, of cyclohexanone 1,2-propanediol ketal is 80.1%, of butanone ethylene ketal is 69.5%, of butanone 1,2-propanediol ketal is 78.5%, of 2-phenyl-1,3-dioxolane is 56.7%, of 4-methyl-2-phenyl-1,3-dioxolane is 86.2%, of 2-propyl-1,3-dioxolane is 87.5%, of 4-methyl-2-propyl-1,3-dioxolane is 87.9%.

Objective To analyze the causes of severe adverse reaction after painless abortion and to offer some advices for prevention and cure.Methods Clinical files and results of 3 patients with severe adverse re-action after painless abortion were analyze.Results Total of 130-140 mg· kg -1 propofol intravenous injection and oxytocin 10 u intramuscular injection was used in three patients underwent painless abortion.Three patients have the symptoms of chills , fever , nausea , vomiting , low blood pressure , damages of multiple organs and elevation of peripheral white blood cell and neutrophilic granulocyte.The shock occurred within 0.5-4 h after operation.Conclusion The symptoms of severe adverse reac-tion after transfusion are very serious.Early diagnosis and rational man-agement are the key to the successful salvage.

OBJECTIVETo study the genetic diversity of C. morifolium on the chemical constituents.

METHODChemical constituents of four cultivars cultivated with the same conditions were compared in three types of index: chlorogenic acid, flavonoid and volatile oil.

RESULT AND CONCLUSIONWith different cultivars and processing methods, the contents of chlorogenic acid, flavonoid and volatile oil extracted from C. morifolium vary great extent.

Chlorogenic Acid ; analysis ; Chrysanthemum ; chemistry ; genetics ; growth & development ; Flavonoids ; analysis ; Flowers ; chemistry ; genetics ; Hot Temperature ; Oils, Volatile ; analysis ; Plants, Medicinal ; chemistry ; genetics ; growth & development ; Polymorphism, Genetic ; Quality Control ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the prognostic factors of small cell lung cancer (SCLC) and establish a reliable model of clinical prognostic index.

METHODSKaplan-Meier and Cox regression were used to analyze the relationship between survival time and prognostic factors in 60 cases of SCLC. The prognostic factors included clinical and laboratory parameters, serum cytokeratin fragment 19 (CYFRA21-1), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).

RESULTSKaplan-Meier analysis showed that poor prognosis was in patients with KPS < 80 or extensive disease and unrelated to other clinical parameters such as age, sex and smoking index, and in patients with serum NSE > 30 micro g/L, CEA > 5.0 micro g/L, CA125 > 37 KU/L and sIL-2R > 500 KU/L. Serum IL-2 and CYFRA21-1 were also elevated, but had no significant prognostic value. Multivariate analysis indicated that serum NSE, stage and treatment of disease were independent prognostic factors. The three prognostic factors enabled establishment of a prognostic index (PI) based on a simple algorithm: PI = NSE (0 if < or = 30 micro g/L, 1 if > 30 microg/L) + stage (0 = LD, 1 = ED) + CEA (0 if < or = 5.0 microg/L, 1 if > 5.0 microg/L).

CONCLUSIONThe stage of disease, systemic treatment and the level of serum NSE are independent prognostic factors. Without considering the influence of treatment-related factors on survival, the levels of serum CEA, NSE and stage of disease before treatment are significant independent prognostic factors. PI calculated on the basis of CEA, NSE and stage is recommended to predict the survival of SCLC.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Brain Neoplasms ; secondary ; Carcinoma, Small Cell ; mortality ; secondary ; therapy ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; secondary ; Lung Neoplasms ; mortality ; pathology ; therapy ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Survival Rate

OBJECTIVETo explore the expression of vascular endothelial growth factor (VEGF) and its receptors (flt-1 and flk-1) in the retina of retinopathy of prematurity (ROP), and its relation to the alteration of retinal blood vessels.

METHODSEighty-six newborn Sprague-Dawley rats were randomly divided into hyperoxia and air groups, then each group was further divided into 1, 3, 7 and 14 days subgroups. The rats in hyperoxia group inhaled 75% oxygen and ROP model was thus set up. These animals were sacrificed respectively after 1, 3, 7 and 14 days, then the retinal endothelial cells were marked by CD34 to observe the change of retinal blood vessels. The expression of VEGF, flt-1 and flk-1 in the retina was measured by immunohistochemistry.

RESULTSThe retinal capillary density index (RCDI) in control group increased as days went on (F = 21.589, P < 0.01, but it was the least on the 7th day in hyperoxia group, after the rats had been returned to air for 7 days, RCDI increased significantly (F = 67.885, P < 0.01); In the control group, the expression of VEGF and flk-1 was the strongest in the retina on the 7th day, the result had significant difference as compared with the 1st and 14th day (P < 0.05). The expression of VEGF and flk-1 on the 7th day in hyperoxia group was weaker than that of control group (P < 0.05). But on the 14th day in hyperoxia group, they were stronger than that of control (P < 0.05). The localization of the expression of flt-1 was changed when blood vessels altered, but there was no significant difference in expression intensity as a whole (P > 0.05).

CONCLUSIONWhen the premature retina was exposed to hyperoxia, the expression of VEGF and flk-1 was reduced, and retinal blood vessels were also decreased; but the expression of VEGF and flk-1 was stronger in retina when premature rats were exposed to relative hypoxia, and the retinal blood vessels also increased significantly. It is concluded that VEGF and flk-1 may play important roles in the development of retinal blood vessels and its change in ROP. However, flt-1 has less effect compared with flk-1.

Animals ; Animals, Newborn ; Disease Models, Animal ; Female ; Hypoxia ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Vascular Endothelial Growth Factor ; analysis ; Retina ; chemistry ; pathology ; Retinal Diseases ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo explore the efficacy and safety of endoscopic submucosal enucleation (ESE) for gastric submucosal tumors (SMTs) originated from muscularis propria.

METHODSA total of 116 patients with gastric SMT originated from muscularis propria underwent ESE in Department of Gastroenterology of the Taizhou Hospital between July 2006 and March 2011. The occurrence of intra-operative and post-operative complications and corresponding treatment were recorded. After the treatment of ESE, the patients were followed up endoscopically.

RESULTSThe success rate of operation was 96.6%. The mean time of the procedure was (51.9±16.3) min. Complications included intra-operative bleeding (n=9, 7.8%), perforation (n=20, 17.2%), and post-operative bleeding (n=3, 2.6%). Among them, 5 cases (4.3%) required surgical intervention. None of patient had other complications such as peritoneal abscess or peritonitis. The mean hospitalization time after ESE was 6.1 days. The median follow-up period was 12 months (range, 3-48 months) and there was no residual tumor or recurrence.

CONCLUSIONESE is a safe and feasible treatment for patients with gastric SMT originated from muscularis propria.

Adult ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Gastroscopy ; methods ; Humans ; Male ; Middle Aged ; Stomach Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo construct cDNA subtracted libraries from gastric dysplasia and further screen differentially expressed genes.

METHODSRelatively pure dysplasia and normal tissue were procured by manual microdissection, and amplified by cDNA-PCR, which was used to carry on for suppression subtractive hybridization (SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Differentially expressed fragments were verified by dot hybridization.

RESULTSTwo subtracted cDNA libraries were constructed. Among 26 sequenced clones, 15 fragments corresponded to known genes, 3 fragments were known EST and 8 fragments were unknown EST (GenBank BQ164614-BQ164616, BQ291516-BQ291520). Fifteen fragments were verified to be differentially expressed in gastric dysplasia.

CONCLUSIONSSubtracted cDNA libraries from gastric dysplasia are constructed using combination of microdissection-cDNA PCR and SSH setup in our laboratory. Some fragments have been screened and verified to help to search for novel associated genes with gastric carcinogenesis.

Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Microdissection ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; Precancerous Conditions ; genetics ; pathology ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics ; pathology

Objective To investigate the effects of Yiqi Huoxue Xiaozheng Formula on cell apoptosis in rats with benign prostatic hyperplasia (BPH); To preliminarily explore its mechanism of action. Methods Totally sixty 8-week-old male SD rats were randomly devided into normal group, positive control group, Yiqi Huoxue XiaozhengFormula low-, medium-, and high-dose groups, with 10 rats in each group. Except for the rats in normal group, rats in other groups were treated with castrate and testosterone propionate injection to replicate BPH model. The normal group and the model group were given normal saline for gavage; Yiqi Huoxue Xiaozheng Formula low-, middle-, and high-dose groups were given Yiqi Huoxue Xiaozheng Formula 10, 20, 40 mg/100 g for gavage, respectively; rats in positive control group were given Longkai Granule suspension 11 mg/100 g for gavage, once a day, for 30 d. Twenty-four hours after the last administration, the rats were sacrificed by cervical dislocation. The prostate tissue was taken. The protein expressions of Bcl-2 and Bax in the prostate tissue were detected by immunohistochemistry. Results Compared with the normol group, the expression of Bax in the prostate tissue of the model group decreased significantly, while the expression of Bcl-2 increased significantly; Compared with the model group, the expression of Bax in the prostate tissue of rats in administration groups increased significantly, while the expression of Bcl-2 was significantly reduced, and Yiqi Huoxue Xiaozheng Formula high-dose group was superior to other groups. Conclusion Yiqi Huoxue Xiaozheng Formula can improve benign prostatic hyperplasia by promoting the apoptosis of rat prostate tissue, and its mechanism is related to the increase of the expression of Bax protein and the decrease of Bcl-2 protein expression in prostate tissue.