According to medicinal chemistry experiment characteristics,the teaching meth-ods and grading system were reformed,The main point of this course is improving students’ex-perimental technique and skill capability so as to effectively arouse the students’interest in study and improve their comprehensive experimental ability.

In order to identify rat ovarian germ cells, we expressed and purified rat RVLG protein in Escherichia coli cells and prepared a mouse anti-rat RVLG polyclonal antibody. The rat RVLG cDNA was obtained from rat testicle tissue by RT-PCR and was cloned into the vector pMD19-T. Sequence analysis proves that the cloned RVLG cDNA fragment was 60 bp longer than that released in the GenBank (NM_001077647), resulting from an alternative splicing of the RVLG pre-mRNA. The RVLG cDNA was double digested with the restriction endonucleases BamH I and EcoR I, and then was extracted from gel and inserted into the prokaryotic expression vector pGEX-4T-1. The recombinant expression plasmid pGEX-RVLG was verified for successful construction and then was transformed into Escherichia coli BL21(DE3) for induction to express the GST-RVLG fusion protein by IPTG. The GST-RVLG fusion protein was expressed in Escherichia coli BL21 (DE3) at a high level which accounts for more than 10% of the total bacterial cellular protein. The purified RVLG protein was used as an antigen to immunize KM mouse for the production of polyclonal antibody in ascetic fluid followed by celiacly injecting the mouse with S180 cells. The mouse anti-rat RVLG antibody was analyzed by ELISA, Western blotting and immunohistochemistry for its specificity and titer. The antibody could recognize RVLG protein specifically and its titer was about 1:20 000. These results confirm that the mouse anti-rat RVLG polyclonal antibody with high affinity and specificity has been prepared successfully, and lay a foundation for our ongoing research on the specific expression of RVLG in rat ovary.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Base Sequence ; Cloning, Molecular ; DEAD-box RNA Helicases ; biosynthesis ; genetics ; immunology ; DNA, Complementary ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Female ; Mice ; Molecular Sequence Data ; Ovary ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

To study alkaloid constituents in Corydalis decumbens,thirteen compounds were isolated from 95％ ethanol extracts of Corydalis decumbens (Thunb) Pers.by silica gel,RP-C1s,Sephadex LH-20 column chromatographer,recry stallization,thin-layer chromatography and HPLC.Their structures were elucidated on the basis of spectral data combined with physiochemical properties as tetrahydropalmatine (1),oxyhydrastinine (2),doryanine (3),palmatine (4),bicuculline (5),canadine (6),tetrahydrocoptisine(7),corydaldine (8),epicorynoxidine (9),N-methylcorydaldine (10),(+)-corlumine (11),N-methyl-6,7-dimethoxyisoquinolone (12) and oxysanguinarine (13).Compounds 2,3,6,7,and 9-13 were isolated from this plant for the first time.In addition,compounds 2,3,and 9-13 were obtained firstly from this genus.Pharmacological experiments showed that tetrahydropalmatine (1) might have analgesic or sedative effects,and the bicuculline (5) could probably induce epilepsy.

Objective To summarize the magnetic resonance imaging (MRI) features of the persistcnt perineal sinus (PPS).Methods The retrospective and descriptive study was conducted.The clinical data of 7 patients with PPS who were admitted to the Nanjing University of Traditional Chinese Medicine between July 2010 and January 2017 were collected.Patients received horizontal,anteroposterior axes and coronal scanning of MRI after abdominoperineal resection (APR) of rectal cancer.Two physicians read collectively films and then achieved consistent results if there was a disputed result.Observation indicators:(1) MRI features;(2) treatment and follow-up situations.Patients underwent resection of PPS according to results of MRI examination,and then regular pathological examination.Follow-up using telephone interview was performed to detect sinus recurrence and reoperation up to April 2017.Results (1) MRI features:① Lesion location:lesions of PPS in 7 patients were located at the presacral areas,top side was up to the third sacral plane,and lower side extended down to the pubic area.② Morphology and signal:sinus tract of 7 patients showed irregular flaky and tubular abnormal signals;fibrotic sinus tract wall showed low signals in T1 weighted imaging (T1WI) and T2WI;contents of sinus tract in 7 patients showed low signal in T1WI and high signals in T2WI and T2WI-fat suppression (T2WI-FS).Of 7 patients,uniform signal and mixed signal were respectively detected in 2 and 5 patients.③ Branches and cystic spaces:1 patient had a branch at the tip of the coccyx,extending to the rear of the sacral vertebra.The top of sinus tract in 4 patients enlarged to form a cavity.④ Signs of infection:signs of infection in 3 patients included blurred and exuded fat gaps around the sinus tract,unclear and partially adhesion with pelvic organ;the pelvic floor muscles in 1 patient were thickened and swollen,with a high signal in T2WI-FS;flaky fluid shadows in 2 patients were seen in the posterior sinus,with a high signal in T2WI-FS,showing a tissue edema of anterior sacrum.⑤ Enhancement features:result of MRI examination of 1 patient showed mild and heterogeneous enhancement of sinus.(2) Treatment and follow-up situations:among 7 patients undergoing sinus resection,2 received curettage of superior sinus due to high location,and 5 received successfully operation.All patients underwent postoperatively regular antibiotic therapy.The healing time was 14-78 days,with an average time of 42 days.The postoperative pathological examination showed different degrees of inflammatory cell infiltration.Seven patients were followed up for 2-74 months,with an average time of 40 months.Of 7 patients,sinus in 6 patients was healed,1 had recurrence of sinus and didn't receive therapy.Seven patients didn't undergo debridement or transposition of skin flap again.Conclusion The MRI imaging features of PPS includes flaky and tubular abnormal signal in the the anterior sacrum,low signal in T1WI,high signals in T2WI and T2WI-FS,and the enhanced scanning of sinus duet shows mild and heterogeneous enhancement.