Teaching practice of Experimental Nuclear Medicine for undergraduates of basic medicine by literature reading / 中华医学教育探索杂志

In order to improve the students' interest and cultivate their scientific literacy, we tried to combine literature reading with the teaching material content of Experimental Nuclear Medicine. To complete literature interpretation, the students performed intensive reading and overall report, and the teacher made a final review and summary. The novel teaching method could improve the understanding of theoretical knowledge, deepen the recognition of experimental nuclear medicine technology, and expand the scientific view for the students, which are beneficial to cultivate the scientific literacy of the undergraduates.

Analyzed the distribution of pathogenic spectra and antibiotic resistance by isolating from 4 238 blood cultures / 国际检验医学杂志

Objective To explore the distribution features of pathogenic spectra and antibiotic resistance of the isolates from blood cultures in hospital from June 2012 to June 2016.Methods A total of 4 238 blood samples from June 2012 to June 2016 were evaluated by BD Bactec FX-200,the identification results were used for retrospective analysis.Results A total of 455 positive pathogens were isolated from 4 238 blood cultures sample,the positive rate was 10.74％,Gram-positive accounts for 38.02％,Gram-negative bacilli accounts for 60.00％,Fungi accounts for 1.98％.Positive pathogens were distributed in newborn baby and middle-older patients,accounting for 6.78％ and 76.17％respectively.Which the Enterobacteriaceae accounting for 54.10％,the major consists were Escherichia coli and Klebsiellapneumoniae;Non-fermentative bacterial which consists of Pseudomonas aeruginosa and Acinetobacterbaumannii accounting for 2.90％.The major pathogens in Gram-positive cocci was Staphylococcus,accounting for 25.87％.Enterobacteriaceae were more sensitive to Meropenem,Imipenem and so all.Non-fermentative bacterial were more sensitive to Piperacillin/Tazobactam.Staphylococcus were more sensitive to Vancomycin and Linezolid.Streptococcus were sensitive to Vancomycin.Conclusion Combined with the distribution features of pathogenic spectra and antibiotic resistance,clinicians should pay attention to use of drugs reasonably to enhance the cure rate of bacteremia and Fungalemia.

Effect of dihydroartemisinin on cell proliferation and radiotherapy sensitization of lung cancer cell H1299 / 中国免疫学杂志

Objective:To investigate the inhibitory effect of dihydroartemisinin on non-small cell lung cancer H1299 cells.Methods:Through the CCK-8 method for determining the IC 10 of dihydroartemisinin ,choose low dose IC 10 as the experimental concentration,CCK-8 to observe artemisinin in low doses of H1299 cell proliferation, cycle and the influence of radiation sensitivity.Results:IC10 of dihydroartemisinin was 14.87 μmol/L,dihydroartemisinin could inhibit proliferation of H 1299 cells,slow down the cell cycle , and increased the radiation sensitivity.Conclusion: Dihydroartemisinin can inhibit cell proliferation , cell cycle arrest in S phase ,increase the radiation sensitivity.

Inhibitory effect of ¹³¹I-CD133mAb combined with cisplatin on liver cancer cells in vitro and in a tumor-bearing mouse model / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the inhibitory effect of CD133 monoclonal antibody labeled with ¹³¹I (¹³¹I-CD133mAb) on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft.</p><p><b>METHODS</b>¹³¹I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with ¹³¹I-CD133mAb plus cisplatin (DDP), ¹³¹I -CD133mAb, DDP, or no treatment (blank control) were examined for cell proliferation suppression by MTT assay with the IC₅₀ calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with ¹³¹I -CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining.</p><p><b>RESULTS</b>The labeling ratio of ¹³¹I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. ¹³¹I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice.</p><p><b>CONCLUSION</b>¹³¹I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.</p>

Screening and primary identification of anti-CCR7 scFv from large phage library / 中国药理学通报

Aim To screen and identify anti-CCR7 single chain fragments variable(scFv)from lager phage library and to detect the scFv efficiency.Methods The insert ratio of ScFv antibodies library was identified by PCR.The products digested by Sfi I/Not I double enzyme.Panning against breast cancer cell and CCR7 were performed four and three rounds respectively.Positive clones were transformed to E.coli HB2151, and their dissolvability was assayed.The soluble scFv was purified by affinity chromatography, and its relative molecular mass was determined by Western blot.The ELISA assay was used to identify the immunocompetence of the antibody.Immunocytochemical staining and radioimmunoimaging were employed to determine the affinities of scFv binding with CCR7 in cell line and in nude mice.Results The insert ratio of ScFv gene was 90%(18/20), enzyme digest reaction showed the aim products on 1% agarose gel.ScFvs were obviously enriched after 7 round panning.Western blot result showed soluble scFv's molecular mass was about 34 KD.ELISA analysis showed dissolved antibody had high immunocompetence to MDA-MB-435 s cells.Immunocytochemical staining and radioimmunoimaging indicated that ScFvs were bound efficiently to MDA-MB-435 s cells which expressed CCR7.Conclusions ScFvs against CCR7 are successfully acquired by screening the phage antibody library.The soluble ScFvs have high affinity and specifical binding to human breast cancer cells.

Screening and identification of human anti-HIF-1? scFv of lung adenocarcinoma / 第三军医大学学报

Objective To screen human anti-hypoxia-inducible factor (HIF)-1? scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones. Methods Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1? was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis. Results After panning,the target scFv was enriched,and ELISA results showed that positive reactions to HIF-1? were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1? scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30?103. The binding activity and specificity were confirmed by ELISA. Conclusion Human anti-HIF-1? scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.

Experimental study of epidermal growth factor receptor imaging in glioma / 重庆医科大学学报

Objective:To explore the feasibility of diagnosing glioma which expressing high level EGFR by 131I-EGF.Methods: In vitro cellular uptake and cellular retention experiment of ~(131)I-EGF was carried out with C6 cell.Radiodistribution experiment was carried out after injection of ~(131)I-EGF into 2-week-old tumor bearing Wistar rats,at 30min,2 hour,6 hour and 12 hour. SPECT images were carried out after 12 hours of intravenous injection of ~(131)I-EGF between tumor bearing rats and normal rats. Results:The rate of C6 cellular uptake and retention in vitro was high.At all selected time,~(131)I-EGF was mainly distributed in liver,spleen and kidney,partially distributed in tumor,but the radioactivity was stronger and stronger with the time passed,until reached the highest strong at 12h.Tumor tissues of tumor bearing rats were seen clearly by SPECT imaging after 12 hours of intravenous injection of ~(131)I-EGF,the liver and spleen were the major normal organs visualized on the images,and the other tis- sues didn't be seen obviously.Conclusions:~(131)I-EGF could offer a new and specific approach to diagnose tumor such as glioma and other tumors expressing high level EGFR from molecular level.

Construction and screening of human-originated phage single-chain antibody library associated with esophageal cancer / 中国药理学通报

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

The recovery course of pituitary-thyroid axis after ~(131)Ⅰin hyperthyroidism / 重庆医科大学学报

Objective:To review the recovery course of pituitary-thyroid axis by follow-up of hyperthyroidism patients with ~(131)Ⅰ therapy.Methods:This study involved 322 patients undergoing~(131)Ⅰ therapy between 1997 and 2000 in our hospital.Serum T_3,T_4 and TSH were regularly evaluated for 3 years after therapy.Results:Our study showed that serum T_3,T_4 and TSH returned to normal levels step by step,but the recovery of TSH fell behind that of T_3 and T_4.Conclusion:Our study demonstrates that clinical manifestations disappear in 1 year after ~(131)Ⅰ therapy,while the recovery of pituitary-thyroid axis takes more time.