Objective To investigate of the radiological manifestations in congenital cystic adenomatoid malformation (CCAM) of lung in adult and to improve the diagnostic accuracy of CCAM of lung in adult.Methods Five cases with pathologically proved CCAM of lung in adult were retrospectively analyzed.Chest X-ray was available in 5 cases and chest CT was performed in 2 cases.Results On plain chest radiography,thin wall air cystic lesions with air-fluid level were detected in 3 cases.Honeycomb like small cystic lesion was detected in 1 case.Multiple round cystic lesions were found in 1 case.CT scan of the chest demonstrated a round thin wall air cystic lesion in the lateral segment of right lung's middle lobe,and a thin wall air cystic lesion with the wall merged into the shadow of chest wall in the left apicoposterior segment in one case.Small cystic lesions just like honeycomb were found in bilateral basal segments of the inferior lobes,with a 0.8—1.0 cm sized round mass revealed in the right lung′s inferior lobe basal segment, and the mass was spiculated in another case.Conclusion The imaging signs of CCAM of lung in adult is cyst or cyst-solid and at the risk of developing carcinoma.

To obtain the key genes for chlorogenic acid biosynthesis of Lonicera hypoglauca, four new genes ware obtained from the our dataset of L. hypoglauca. And we also predicted the structure and function of LHPAL4, LHHCT1 , LHHCT2 and LHHCT3 proteins. The phylogenetic tree showed that LHPAL4 was closely related with LHPAL1, LHHCT1 was closely related with LHHCT3, LHHCT2 clustered into a single group. By Real-time PCR to detect the gene expressed level in different organs of L. hypoglauca, we found that the transcripted level of LHPAL4, LHHCT1 and LHHCT3 was the highest in defeat flowers, and the transcripted level of LHHCT2 was the highest in leaves. These result provided a basis to further analysis the mechanism of active ingredients in different organs, as well as the element for in vitro biosynthesis of active ingredients.
Chlorogenic Acid ; metabolism ; Cloning, Molecular ; Computational Biology ; Gene Expression Regulation, Plant ; Lonicera ; chemistry ; classification ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary

Two hundred and sixty-one subjects were recruited from in-patients and subjects for phaysical Check-up,and were divided into normal control group (NC),nonalcoholic fatty liver disease group (NAFLD),type 2 diabetes mellitus group (T2DM),and T2DM accompanied by NAFLD group (DMN).According to the result of ultrasonic examination,the patients with T2DM were further divided into non-NAFLD group,light fatty liver group (NAFLD-L group),moderate fatty liver group(NAFLD-M group),and severe fatty liver(NAFLD-S group).Fasting plasma glucose,blood lipid,liver function,kidney function,and serum retinol-binding protein 4 (RBP4) levels were determined.The risk of various indicators for NAFLD was determined by correlation analysis and logistic regression analysis.The results showed that fasting glucose levels in diabetics with or without NAFLD were significantly higher than those in NC and NAFLD groups(P＜0.01).Triglyceride (TG) level in DMN group was significantly higher than those in other three groups(all P＜0.01),while high density lipoprotein-cholesterol level was lower than those in other three groups(all P＜0.01).Systolic blood pressure and diastolic blood pressure in DMN group were higher than those in NC and T2DM groups (P＜0.05 or P＜0.01).Serum RBP4 level in patients with NAFLD was significantly higher compared with the subjects without NAFLD [45.00 (38.75,51.00) mg/L vs 51.00 (43.00,62.00) mg/L,P ＜0.01],and was rising with the progress of NAFLD [NAFLD-L group 44.00 (37.00,51.00) mg/L,NAFLD-M group 52.00(46.00,63.00) mg/L,and NAFLD-S group 78.5 (72.75,83.00) mg/L,all P＜0.01].Logistic regression analysis showed that the RBP4 level was an independent factor associated with NAFLD (P =0.029).In addition,serum RBP4 level was correlated with body mass index,waist-to-hip ratio,serum gamma-glutamyl transpeptidase,total cholesterol,TG,aspartate aminotransferase,alanine aminotransferase,prealbumin,creatinine,blood urea nitrogen,and uric acid.These resuhs suggest that serum RBP4 is an independent risk factor of NAFLD.

This study aims to construct a eukaryotic expression system for envelope gene of Jaagsiekte sheep retrovirus, observes its localization in 293T cells, and investigates the potential in inducing malignant transformation of NIH3T3 cells. By RT-PCR, the full-length cDNA of envelope gene of Jaagsiekte sheep retrovirus (exJSRV-env) was amplified from the extract of naturally infected sheep lung. The clone of target gene was sub-cloned into eukaryotic expression system pEGFP-C1, and validated by PCR, restriction endonuclease, and sequencing. Bioinformatic analysis concerning biological function and cellular localiza tion of exJSRV-env was also performed. The recombinant clone of exJSRV-env was transfected into 293T cells and NIH3T3 cells by Lipofectamine LTX. The expression and celluar localization in 293T cells were validated by confocal microscopy. Soft agar colony formation assay was employed to test the anchorage-independent growth of NIH3T3. DNA sequencing and restriction enzyme digestion with Kpn I and Hind III indicated the correct construction of the recombinant plasmid, which was named pEGFP-C1/exJSRV-env. Amino acid sequence alignment of exJSRV-env with reference sequences found 85%-100% homogeneity. A YRNM motif was discovered at the cytoplasmic tail of envelope gene, which is exclusively found in exogenous viruses. Phylogenetic tree analysis showed that our clone of exJSRV-env clustered closely with pathogenic exogenous Jaagsiekte sheep retroviruses. Fluorescence microscopy indicated typical membrane localization of exJSRV-env protein. NIH3T3 cells transfected with exJSRV-env lost contact inhibition, and acquired colony forming ability in soft agar. This study indicated that envelope protein of Jaagsiekte sheep retrovirus can induce malignant transformation of mouse fibroblast cell NIH3T3. Discoveries of this study provide a basis for further structural and functional research on Jaagsiekte sheep retrovirus envelope protein.
Amino Acid Sequence ; Animals ; Betaretrovirus ; chemistry ; classification ; genetics ; physiology ; Cell Transformation, Viral ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Phylogeny ; Retroviridae Infections ; veterinary ; virology ; Sequence Alignment ; Sheep ; Sheep Diseases ; virology ; Transformation, Genetic ; Tumor Virus Infections ; veterinary ; virology ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism

Objective To construct the stable stathmin-overexpression SMMC-7721 hepatocellular carcinoma cells and to explore the effect of stathmin-overexpression on the cell proliferation and metastasis in SMMC-7721 cells .Methods By using liposome , Flag-pcDNA3 .1 and Flag-pcDNA3 .1-stathmin plasmid were transfected into SMMC-7721 cells respectively ,the stable Flag-pcD-NA3 .1 expression cells(control group) and the stable stathmin-overexpression cells(experimental group) were established after an-tibiotic resistant gene screening ,and the cell lines were identified by Western Blot .Subsequently ,the cell proliferation was detected by cell count kit(CCK-8) and the soft agar assay ,the apoptosis and cell cycle were determined by the flow cytometry (FCM ) ,and the cell motility and invasion were analyzed by the Transwell assay in vitro .Results The stathmin protein expression of the experi-mental group was significantly increased compared with the control group (0 .76 ± 0 .12 vs .0 .16 ± 0 .05 ,P<0 .05) ,which indicated that the stathmin-overexpression human SMMC-7721 hepatocellular carcinoma cell line was successfully constructed .CCK-8 and the soft agar assay showed that the cell proliferation of the experimental group was higher than that of the control group (0 .29 ± 0 .03 vs .0 .60 ± 0 .05 ,P< 0 .01);additionally ,the apoptotic ratio of the experimental group was decreased compared with the control group[(11 .57 ± 1 .09)% vs .(5 .80 ± 0 .33)% ,P<0 .05] ,the cell cycle was arrested in the stage G2/M ;the Transwell experiment results verified that the cell motility and the invasive ability of the experimental group were obviously reinforced compared with the control group[transmenbrane cell numbers in migrant assay :(54 .03 ± 7 .21) vs .(130 .45 ± 14 .13);transmenbrane cell numbers in invasive assay :(17 .75 ± 2 .52) vs .(57 .76 ± 8 .50) respectively] ,the differences had statistical significance(P<0 .01) .Conclusion The overexpression of stathmin promotes the cell proliferation and the invasive ability in SMMC-7721 hepatocellular carcinoma cells .

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

This article introduces an overview of management system for conflict of interest of the European Pharmacopoeia Commission (EPC) and the United States Pharmacopoeia Convention (USP). The EPC and USP have standardized the management system for conflict of interest in drug standard work in multiple management documents, such as the Guide for the Work, Code of Practice for the Work, Form for Declaration of Interests and Confidentiality Undertaking of the EPC, bylaws, Rules and Procedures of the Council of Experts, Code of Ethics, Standards of Conduct of the USP, in order to ensure the transparency and fairness of drug standard development, improve the credibility and rigor of drug standards. This article introduces the management system for conflict of interest of the EPC and USP, providing reference for the improvement of relevant management systems of the Chinese Pharmacopoeia Commission.

Objective To explore the pulse diagram parameter changes of chronic renal insufficiency patients with five symptoms types(spleen kidney qi deficiency ,spleen kidney Yang deficiency ,kidney liver Yin deficiency and the deficiency of Yin and Yang ) , and to establish the differentiation mode of each symptoms type for assisting the clinical diagnosis .Methods The DS01-C pulse manifestation instrument made by the Shanghai Daosh company was adopted to detect and analyze the pulse manifestations in the healthy control group and the chronic renal insufficiency group .Results The healthy control group was dominated by the normal pulse manifestation .The chronic renal insufficiency group was dominated by the taut pulse and its concurrent pulse .Along with the progress of the disease ,the pulse manifestations also appeared the corresponding changes .The patients with spleen kidney qi defi-ciency and spleen kidney Yang deficiency were dominated by the taut pulse .Comparing the patients with liver kidney Yin deficiency and Qi Yin deficiency ,the taut pulse and concurrent rapid pulse were common ,in addition ,the former also had the deep pulse .The patients with Yin and Yang deficiency showed the slow pulse and the taut pulse or the taut pulse and rapid pulse .Conclusion The pulse manifestation change in the patients with chronic renal insufficiency is dominated by the taut pulse and the concurrent pulse , the pulse manifestation change of various symptoms types are complex .

Objective To investigate the application of polymerase chain reaction and single strand conformation polymorphism analysis(PCR-SSCP)to the screening of gene mutation of exon 13 of the LDLR gene in familial hypercholesterolemia(FH).Methods Peripheral blood DNA of 16 clinically diagnosed FH patients was extracted and the exon 13 coding region of the LDLR gene was amplified by PCR.PCR products were separated by optimized SSCP electrophoresis and visualized by silver staining.DNA fragments with abnormal mobility were sequenced to determine the nature and position of mutations.Results The SSCP electrophoresis conditions were optimized as 8%polyaerylamide(degree of cross linking 49:1)gel without glycerin at a electrophoresis temperature of 10℃ or 8%polyacrylamide gel with 5%glycerin at room temperature,gel thickness of＜0.4 mm,and a voltage of 5 V/cm.DNA fragments were well resolved with the conditions and sequencing of the abnormal bands resuhed in detections of missense mutations of A606T,D601N,Y601D and G636V together with a synonymous mutation of 1959C→T in 4 patients and a sole synonymous mutation of 1959C→T in other 4 patients.Conclusion PCR-SSCP is an effective method for the screening of exon13 mutations of LDLR gene in FH patients.

To investigate the anticancer effects of ring C in 18β-glycyrrhetinic acid (GA), a series of GA derivatives featured with 9(11)-ene moiety in ring C were designed and synthesized. The structures were confirmed by IR, LC-MS and 1H NMR. Their inhibitory effects towards human prostate cancer PC-3 and leukemia HL-60 cell lines were determined. Most of the derivatives displayed stronger antiproliferative activities than GA. Particularly, compound 14 showed promising anticancer activity with the GI50 values of 4.48 µmol · L(-1) and 1.2 µmol · L(-1) against PC-3 and HL-60 cells respectively, which is worth further study.
Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Drug Design ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; HL-60 Cells ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; pathology