Total RNA was isolated from Trichomonas vaginalis and Tv-Sir2-like cDNA was amplified by RT-PCR and cloned into pGEM-T Easy plasmid. A fragment of Tv-Sir2-like cDNA was subcloned into the expression vector pET-41b and expressed in E.coli BL21 with induction of IPTG. The full-length of Tv-Sir2-like cDNA was cloned and sequenced. The prokaryotic expression system of pET-41b/Tv-Sir2-like was constructed. The fusion protein of Tv-Sir2-like was expressed in E.coli BL21, occupying 30% of the total bacterial protein after being induced by IPTG for 5 h. SDS-PAGE analysis showed that the fusion protein was about Mr 59 000. The recombinant protein of Tv-Sir2-like is efficiently expressed in E.coli BL21.

Objectives The ferredoxins are iron-sulfur proteins, which function in electron transfer reactions in a variety of systems and participate in the activation of the antimicrobial agent metronidazole. The aim of this study is to clone and characterize ferredoxin genes of Trichomonas vaginalis. Methods A cDNA expression library was constructed with T. Vaginalis total RNA. Hundreds of cDNA clones were isolated and sequenced. Sequence analysis was performed using BLAST programs, ClustalW program, etc. Results One of the cDNA clones, which has homology with T.vaginalis ferredoxin, was further analyzed. This cDNA clone has an open reading frame of 312 base pairs. The deduced precursor protein contains 103 amino acid residues with a hydrogenosome targeting sequence (MLSQCSPLRF) at the N-terminal end. The primary sequence analysis revealed that this new ferredoxin (TvFd2) has a high homology (69% identity) to the previous reported T.vaginalis ferredoxin(TvFd). Interestingly, TvFd2 is homologous to both the two subclasses of (2Fe-2S) ferredoxins, the oxidase ferredoxins and the photosynthetic ferredoxins,but with low similarity. The conserved four-cysteine residues, which are predicted to form the iron-sulfur cluster,are arranged in a typical pattern of (2Fe-2S)ferredoxins(-C-X5-C-X2-C-Xn-C-). Conclusion These data show that TvFd2 is a putative new (2Fe-2S) ferredoxin of T.vaginalis. Its biological function remains to be studied.

Objective To investigate microRNA expression profiles of colonic cancer without lymph node metastasis and identify the specific microRNAs associated with carcinogenesis of colon.Methods Cancerous and para-cancerous tissues (5 cm from cancer tissues of the colon) confirmed without lymph node metastasis were collected from 3 patients. The microRNAs were extracted and isolated by mirVana RNA kit. Hybridizations were made by applying the microRNAs to Agilent microRNA microarray. Data analysis was done by software of Feacture Extraction. The discovered microRNAs were confirmed by real time PCR assay. Results Twelve out of 14 microRNAs associated with colonic cancer were up-regulated in colonic tissues. They were miR-106b, miR-135b,miR-18a,miR-18b,miR-196b, miR-19a, miR-224, miR-335, miR-424, miR-20a' , miR-301b and miR-374a. The rest two of miR-378 and miR 378* were down-regulated. Two up-regulated microRNAs (miR-18a and miR-135b) were detected in 3 pairs of cancerous and para-cancerous tissues. The expression level of miR-18a and miR-135b were accordant with the results of real time PCR.Conclusions microRNA.such as miR-106b,miR-135b.miR-18a.miR-18b,miR-196b,ect,were differentially expressed between cancerous and para-cancerous tissues.Many of these target genes are supposed to participate in the process of multiple tunmorgenesis.These microRNAs play an important role in the carcinogenesis of colon.

Metastatic seminal vesicle carcinoma is rare in clinic. The case of renal clear cell carcinoma metastasis to the seminal vesicle gland is more rare, which is difficult to be identified with bladder, prostate and colorectal tumors. In November 2017, 1 case of renal clear cell carcinoma metastasis to the left side of the seminal vesicle gland was admitted in our hospital.Laparoscopy seminal vesicle metastasis tumor resection was performed. Regular bladder irrigation and sunitib oral intake were conducted. No evidence of local recurrence was found within 19 months follow-up.

Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is a rare hereditary disease and characterized by cutaneous leiomyoma, uterine leiomyoma and/or renal cell carcinoma, but rarely associated with vena cava embolism. We treated 1 case of HLRCC syndrome patients with inferior vena cava tumor emboli (Mayo grade Ⅳ), confirmed after genetic testing, the patient and her family refused further treatment. The patient died after two months of follow-up after discharge.