Objective To observe the effect of dihydroartemisinin(DHA) on the ultrastructure of Trichomonas vaginalis cultured in vitro.Methods The trophozoites of T.vaginalis were cultivated with liver extract solution medium containing 1 mg/ml dihydroartemisinin,and were then observed by scanning and transmission electron microscopes after the treatment for 2.5-4.0 h.Results The cell membranes of the trophozoites treated with DHA were damaged considerably.The surface of T.vaginalis showed deepening folds,hollow,and cracks.The nuclear membrane appeared fractures.There were a few of crevices in the nucleus and cytoplasm.Disordered and dilated endoplasmic reticulum,injured and deformed hydrogenosomes were found.Cytoplasm of the damaged parasites spilled over from torn place.After the cell membrane was peeled off,the nuclei,hydrogenosomes and pelta were exposed,which finally resulted in the death of the denatured parasites.Conclusion Dihydroartemisinin can destroy membrane structure and organelles of Trichomonas vaginalis trophozoites,which leads to decomposition and necrosis of the parasites.

AIM To investigate the effect of clonidine on intraocular pressure(IOP) and the possible role in which ? adrenergic mechanism plays. METHODS The change on IOP was observed following clonidine administered via three different routes: (1)clonidine topically administered to eyes; (2)clonidine intracerebroventricularly injected (icv)or topically administered after yohimbine or prazosin icv; (3)microinjection of clonidine into locus coeruleus(LC). RESULTS Clonidine decreased IOP significantly, ? 2 adrenoceptor antagonist, but not ? 1 adrenoceptor antagonist, can reverse the decreasing effect on IOP caused by clonidine icv and administered topically. CONCLUSION Clonidine administered both topically or intracranially can decrease IOP;? 2 adrenoceptor in central nervous system is involved in this effect.

AIM:To investigate the effect of rs 35100176 CCT insertion/deletion polymorphism in the promoter region of importin 8 ( IPO8) gene on its mRNA expression .METHODS:A 342-bp fragment of IPO8 gene promoter con-taining the rs35100176 polymorphism was amplified from 49 DNA samples and sequenced .The IPO8 promoter fragments containing CCT 3-nucleotide insertion or deletion were amplified using the corresponding homozygote DNA samples .The PCR products were sequenced and inserted into the luciferase reporter vector pGL 3-Basic.Recombinant vectors were trans-fected into the cells by Fugene 6.0 and the expression of the reporter gene was detected by a dual-luciferase reporter assay system.The mRNA expression level of IPO8 was detected by real-time PCR in 3-nucleotide insertion or deletion homozygote cells.RESULTS:The sequencing results showed that there were 3 kinds of genotypes in the rs35100176 polymorphism, CCT/CCT, CCT/-and -/-, and the gene frequencies were 18.37%, 55.10% and 26.53%, respectively.The re-combinant expression vectors pGL 3-3N Insertion and pGL3-3N Deletion were successfully constructed .The luciferase assay showed that pGL3-3N Insertion produced significantly lower luciferase activity than that by pGL 3-3N Deletion.Real-time PCR showed that HEK293 cells with 3-nucleotide insertion homozygote expressed relative lower IPO 8 mRNA than Saos-2 cells with 3-nucleotide deletion homozygote .CONCLUSION: The CCT 3-nucleotide insertion variant decreases the pro-moter activity of IPO8, thus affecting the gene expression .

Objective To assess the local treatment efficacy of percutaneous radiofrequency ablation(PRFA) in combination with percutaneous ethanol injection(PEI) therapy protocol for liver metastases in patients with colorectal cancer.Methods From February 2000 to February 2004, total of 61 liver metastatic lesions(1.5～7.8 cm) from 37 colorectal cancer patients were ablated by using RF-2000 generator and 10-needle LeVeen electrode percutaneously guided by B-ultrasound.Tumors larger than 2.5 cm in diameter were treated by multiple overlapping radiofrequency ablations encompassing the entire lesion as well as a rim of normal liver tissue(approximately 1.0 cm).PEI was performed additionally.The Chiba needle and Quadra-Fuse multi-pronged injection needle were used for tumors ranged 1～2 cm and for those larger than 3 cm respectively.Results The procedures were performed successfully in all patients.The post-PRFA/PEI complication rate was 5.4%.The follow-up rate was 97.3%(36/37).Thirteen patients achieved a complete response(CR),and 21 achieved a partial response(PR).The effective rate(CR+PR) was 94.4%.The patients' survival rates were 97.2% at 1 year,75.0% at 2 years,and 52.8% at 3 years respectively.Conclusion The results of this study suggest that the treatment protocol of PRFA in combination with PEI was an effective mini-invasive local ablation therapy,which was part of the comprehensive management of the colorectal liver metastases.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

Objective:To study the change in serum fibroblast growth factor 21(FGF21)level and its correlation with liver functions in elderly patients with hepatitis B virus(HBV)-related cirrhosis.Methods:A total of 150 elderly patients with HBV infection admitted to Beijing Ditan Hospital and China-Japan Friendship Hospital from January 2019 to December 2020 were selected and divided into chronic hepatitis B(CHB)group(n=70)and HBV-related cirrhosis group(n=80). Healthy subjects were selected as the control group(n=50). Serum FGF21 was determined by ELISA method.Clinical data, clinical laboratory indicators and FGF21 as a core parameter of this study were collected and compared among control group, CHB group, HBV-related cirrhosis group.The correlations of FGF21 level with several liver function indexes were analyzed by Pearson correlation analysis.Results:The levels of total bilirubin(TBil)and total biliary acid(TBA)were significantly higher in HBV-related cirrhosis group than in CHB group.The levels of aspartate transaminase(AST), alanine transaminase(ALT), glutamyl transpeptadase(GGT), serum albumin, cholinesterase, FGF21 and HBV-DNA were significantly lower in HBV-related cirrhosis group than in CHB group(all P<0.05). The levels of serum FGF21 were(108.6 ± 7.3)ng/L, (92.5 ± 7.6)ng/L and(75.8 ± 6.3)ng/L in Child-Pugh A, B and C patients, respectively.The level of FGF21 was statistically significantly decreased with the increase of Child-Pugh grading( F=18.290, P=0.000). Serum FGF21 level was positively correlated with AST, ALT, TBil, cholin-esterase and albumin levels( r=0.652, 0.579, 0.609, 0.558, 0.613, all P<0.05). Conclusions:The level of serum FGF21 is significantly decreased in elderly patients with HBV-related cirrhosis, and is positively correlated with liver function indexes.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complemenary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease Sill, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB 1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively.Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.

Objective To investigate the geographic landscape features in Yulong plague natural focus.Methods Four altitudinal gradients (2 400-,2 600-,2 800-,≥3 000 m) were selected as study objects in Yulong plague natural focus from December 2015 to November 2016 to collect and analyze geographic landscape factors (topography and terrain,climate,water areas,vegetation and soil) based on the literatures about this plague focus.Results Yulong plague natural focus was an area with mid-mountain and subalpine topography in northwestern Yunnan Province,and its terrain had the character of lower in the southeast and higher in the northwest.This area belonged to plateau warm temperate monsoon climate of low latitude with an average month temperature 13.6 ℃ from 2005-2016,month air relative humidity averaged 59.4％,and rainfall averaged 884.5 mm a year,which concentrated mostly from June to September.About 98.1％ of the water area in Yulong County was controlled by Chin-sha River.The vegetation and soil were characterized by vertical distribution along with altitude gradients obviously.Conclusion The unique geographical landscape of the plague natural focus in Yulong County is one of important factors affecting the occurrence and epidemic of the plague.