Screening of natural small molecule IDO-1 inhibitors with anti-tumor role / 药学学报

Fragments of the human indoleamine 2,3-dioxygenase 1 (IDO1) gene 5'-UTR (untranslated 1 245 bp region) promoters were amplified by PCR and cloned into pGL4.20 vector in the construction of reporter vector pGL4-IDO1-luc. A549 cells were transfected with the constructed plasmid and IDO1 inhibitor screening model was established with dual-luciferase reporter assay. Based on the model, we screened natural small molecules which could down-regulate the expression of IDO1 on tumor cells. The anti-tumor activities were examined by MTT, Western blotting and lactic dehydrogenase (LDH) release assays. Toosendanin (NS-180) down regulated the IDO1 expression and inhibited IFN-γ-induced STAT1 and STAT3 phosphorylation in A549 cells. Moreover, NS-180 significantly increased the cytotoxicity of co-cultured NK cells on A549 cells in LDH release assays. In summary, NS-180 is a novel and potent IDO1 inhibitor, which has an antitumor activity for cancer immunotherapies.

Constructing a phage-displayed random mutation library of HIV-1 Tat38-61 at the sites of 51 and 55 amino acids in basic region / 生物工程学报

We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.

Investigation of CT guided percutaneous incisional needle biopsy in parapharyngeal region for suspicious nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the safety and efficacy of percutaneous incisional needle biopsy (PINB) in the parapharyngeal region under CT guide for highly suspicious nasopharyngeal carcinoma (NPC) or recurrence of NPC after radiotherapy.</p><p><b>METHODS</b>PINB under CT guide was performed in 32 highly suspicious NPC or recurrence of NPC after radiotherapy through three puncture routes: posterolateral maxillary sinus fatty area, mandibular fossa area, and anterior-mastoid area. Specimens were fixed by 95% alcohol and then underwent pathologic examination.</p><p><b>RESULTS</b>CT guided PINB was successfully performed in every patients with a technical successful rate of 100%. Definitive histopathologic diagnosis was obtained in 30 patients: squamous-cell carcinoma 21, undifferentiated carcinoma 5 and adenocarcinoma 4. The remaining two negative cases were confirmed as fibrosis after radiotherapy. Complications included persistent bleeding of puncture point in one patient and bloody sputum in 3 patients which subsided after symptomatic management. None of these patients was found to have symptoms of nerve injury caused by PINB procedure.</p><p><b>CONCLUSION</b>The CT guided percutaneous incisional needle biopsy in parapharyngeal region through the above three puncture routes for highly suspicious nasopharyngeal carcinoma is safe, rapid and effective.</p>

Hepatitis E virus capsid protein production by high cell density culture of recombinant Escherichia coli / 生物工程学报

Production of Hepatitis E Virus capsid protein by high cell density culture in recombinant E. coli has been studied in 10L and 30L fermentors. The effects of different factors on growth and producing recombinant protein of E. coli have been studied by batch culture, such as different media, the ratio of phosphate and Magnesium sulfate. Comparison of fermentation performance for recombinant E. coli in different fed-methods culture has been investigated by fed-batch culture. The effects of inducing at different stages of growth and time of inducing on growth and producing recombinant protein, also obtained by fed-batch culture. At last, the solubility of inclusion body in different urea concentrations also has been obtained by fed-batch culture. The results show that the concentration of phosphate and Magnesium sulfate in the optimal media is 80mmol/L and 20mmol/L in batch culture respectively, that induction with 1.0mmol/L IPTG at mid log phase (about 45 OD at 600nm) is suitable for growth and recombinant protein expression, the cells were approaching stationary growth phase and the maximum cell OD at 600nm of 80 was achieved in 5h of fed-batch culture, and the expression level is 29.74%. The results also indicate that the solubility of inclusion body in 4mol/L urea solution induced at 37 degrees C reaches 14mg/mL, over 80% inclusion body was resolved. The culture process achieved in 10L fermentor could be successfully scaled up to 30L fenmentor with good reproducibility.

Clinical use of mycophenolate mofetil in renal taransplation / 中国临床药理学与治疗学

Aim To monitor the whole blood trough concentration of cyclosporine A (CsA) in renaltransplant recipients reciving triple therapy with mycophenolate mofetil, cyclosporine andprednisone and to establish an optimal therapeutic window of CsA. Methods Sampleswere measured by specific fluorescence polarization immunoassay. According to the timeafter operation and different therapy plan, the whole blood trough concentration of CsA ineach group was compared with that in control group.Results The optimal therapeuticwindow of CsA with MMF plan was 150~300 ?g? L-1 (less than one month after op-eration), 120~260?g?L-1 (1~