Objective To obtain monoclonal antibody against glypican 3(GPC3) by hybridoma technique and prepare antibody-drug conjugate(ADC) to investigate the anti-tumor activity of ADC.Methods Monoclonal antibody against GPC3with high affinity was obtained by immunizing male BALB/c mice and detected for its affinity to antigen and endocytosis rate in tumor cells by ELISA and flow cytometry.ADC drugs with DAR(drug-to-antibody ratio) of 2 were obtained by sitespecific enzyme coupling technique,and the inhibitory effects on proliferation of tumor cells HepG2 and Hep3b were detected by cell killing experiments.Results Antibody 16C8-8E6 had a high affinity at protein level with the EC_(50) of 2.51 ng/mL,and the affinity at cell level(stable strains 4E1,Hep3b and HepG2 with high expression of GPC3) was significantly higher than that of the original antibody GC33(t=14.9,13.0 and 12.9,respectively,each P <0.05).The fluorescence intensity of stable strain 4E1 with high expression of GPC3 was 20 542±107;The endocytosis rate was better than that of the original antibody GC33,which reached 73.9% within 48 h.16C8-8E6-ADC showed certain inhibitory activity on the proliferation of HepG2 and Hep3b cells.Conclusion The monoclonal antibody targeting GPC3 was successfully obtained,which lays a foundation of the research of immunotherapy targeting GPC3.

Pharmaceutical excipients of animal origin, an important part in pharmaceutical excipients, are widely used in pharmaceutical preparations.However, compared with the pharmaceutical excipients of other origins, pharmaceutical excipients of animal origin have more special requirements in many aspects, such as raw materials, production, quality control, storage, supervision, etc.Chinese Pharmacopoeia 2020 first included the Guideline for Pharmaceutical Excipients of Animal Origin, which introduces the basic ideas and technical requirements for the life cycle quality control of pharmaceutical excipients of animal origin based on the risk management concept.This article illustrates the specificity of the pharmaceutical excipients of animal origin, and interprets the main contents of this guideline in conjunction with relevant domestic and foreign regulations and technical documents, thereby providing comprehensive reference for the implementation of the guideline.

Objective To investigate the changes in the expression of cold-inducible RNA-binding protein (CIRBP) in a radiation-induced lung injury model. Methods Thirty male C57BL/6 mice were randomly divided by body weight into control group (no intervention) and model group (single chest X-ray irradiation with a dose of 20 Gy to build a radiation-induced lung injury model). The mice were dissected five weeks after irradiation. Hematoxylin-eosin staining and Masson staining were used to observe the pathological changes of the lung tissue and the deposition of collagen fibers. Immunohistochemistry was used to measure the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the lung tissue. qRT-PCR was used to measure the expression of CIRBP mRNA in the lung tissue. The expression of CIRBP protein in the lung tissue was determined by the immunofluorescence assay and Western blot. Results Compared with the control group, the model group showed significant pulmonary vascular congestion, significant inflammatory cell infiltration, significant thickening of some alveolar septa, significantly increased IL-6 expression [(129.41 ± 5.58) vs (187.22 ± 34.77), t = 3.179, P < 0.05], significantly increased TNF-α expression [(137.52 ± 23.53) vs (187.02 ± 19.16), t = 5.069, P < 0.05], significantly increased CIRBP mRNA expression [(1 ± 0.08) vs (1.97 ± 0.39), t = 3.45, P < 0.05], and significantly increased CIRBP protein expression [(9.32 ± 1.26) vs (14.76 ± 1.61), t = 3.751, P < 0.05], by the immunofluorescence assay; [(1.13 ± 0.17) vs (1.49 ± 0.14), t = 2.819, P < 0.05], by Western blot). Conclusion The expression of CIRBP is significantly increased in the radiation-induced lung injury model, which may be an important pro-inflammatory factor in radiation-induced lung injury.

Objective: To determine the clinical benefits of internal mammary sentinel lymph node biopsy (IM-SLNB) acquired by breast cancer patients with clinically positive axillary lymph node (ALN), and further optimize the IM-SLNB indications. Methods: All primary breast cancer patients with clinically positive ALN from February 2014 to September 2017 were prospectively recruited in this study. IM-SLNB was performed under the guidance of the modified injection technique. The success rate and visualization rate of IM-SLNB, metastatic rate of internal mammary sentinel lymph node (IMSLN) and its related factors were analyzed, and the clinical benefits were accessed according to the current guidelines. Results: Among 126 patients, all of 94 patients (74.6%) who showed internal mammary drainage successfully underwent IM-SLNB. The incidence of internal mammary artery bleeding and pleural lesion were 4.3%(4/94) and 9.6%(9/94), respectively. The metastatic rate of IMSLN was 38.3% (36/94), which was significantly associated with the number of positive ALN (P<0.001) and tumor size (P=0.024). The lymph node staging of 94 patients who underwent IM-SLNB was more accurate. Among them, 36 cases with positive IMSLN underwent internal mammary radiotherapy (IMRT), while the other 58 cases with negative IMSLN avoided radiotherapy. Conclusions IM-SLNB should be routinely performed in patients with positive ALN. IM-SLNB can provide more accurate staging and guide tailored IMRT to benefit more breast cancer patients.

Objective To determine the clinical benefits of internal mammary sentinel lymph node biopsy (IM?SLNB) acquired by breast cancer patients with clinically positive axillary lymph node ( ALN), and further optimize the IM?SLNB indications. Methods All primary breast cancer patients with clinically positive ALN from February 2014 to September 2017 were prospectively recruited in this study.IM?SLNB was performed under the guidance of the modified injection technique. The success rate and visualization rate of IM?SLNB, metastatic rate of internal mammary sentinel lymph node ( IMSLN) and its related factors were analyzed, and the clinical benefits were accessed according to the current guidelines. Results Among 126 patients, all of 94 patients ( 74.6%) who showed internal mammary drainage successfully underwent IM?SLNB. The incidence of internal mammary artery bleeding and pleural lesion were 4.3%( 4／94) and 9.6%(9／94), respectively. The metastatic rate of IMSLN was 38.3%( 36／94), which was significantly associated with the number of positive ALN (P＜0.001) and tumor size (P＝0.024).The lymph node staging of 94 patients who underwent IM?SLNB was more accurate. Among them, 36 cases with positive IMSLN underwent internal mammary radiotherapy (IMRT), while the other 58 cases with negative IMSLN avoided radiotherapy. Conclusions IM?SLNB should be routinely performed in patients with positive ALN. IM?SLNB can provide more accurate staging and guide tailored IMRT to benefit more breast cancer patients.

Objective To determine the clinical benefits of internal mammary sentinel lymph node biopsy (IM?SLNB) acquired by breast cancer patients with clinically positive axillary lymph node ( ALN), and further optimize the IM?SLNB indications. Methods All primary breast cancer patients with clinically positive ALN from February 2014 to September 2017 were prospectively recruited in this study.IM?SLNB was performed under the guidance of the modified injection technique. The success rate and visualization rate of IM?SLNB, metastatic rate of internal mammary sentinel lymph node ( IMSLN) and its related factors were analyzed, and the clinical benefits were accessed according to the current guidelines. Results Among 126 patients, all of 94 patients ( 74.6%) who showed internal mammary drainage successfully underwent IM?SLNB. The incidence of internal mammary artery bleeding and pleural lesion were 4.3%( 4／94) and 9.6%(9／94), respectively. The metastatic rate of IMSLN was 38.3%( 36／94), which was significantly associated with the number of positive ALN (P＜0.001) and tumor size (P＝0.024).The lymph node staging of 94 patients who underwent IM?SLNB was more accurate. Among them, 36 cases with positive IMSLN underwent internal mammary radiotherapy (IMRT), while the other 58 cases with negative IMSLN avoided radiotherapy. Conclusions IM?SLNB should be routinely performed in patients with positive ALN. IM?SLNB can provide more accurate staging and guide tailored IMRT to benefit more breast cancer patients.

Magnetotactic bacteria (MTB), a group of phylogenetically diverse organisms that use their unique intracellular magnetosome organelles to swim along the Earth's magnetic field, play important roles in the biogeochemical cycles of iron and sulfur. Previous studies have revealed that the bacterial actin protein MamK plays essential roles in the linear arrangement of magnetosomes in MTB cells belonging to the Proteobacteria phylum. However, the molecular mechanisms of multiple-magnetosome-chain arrangements in MTB remain largely unknown. Here, we report that the MamK filaments from the uncultivated 'Candidatus Magnetobacterium casensis' (Mcas) within the phylum Nitrospirae polymerized in the presence of ATP alone and were stable without obvious ATP hydrolysis-mediated disassembly. MamK in Mcas can convert NTP to NDP and NDP to NMP, showing the highest preference to ATP. Unlike its Magnetospirillum counterparts, which form a single magnetosome chain, or other bacterial actins such as MreB and ParM, the polymerized MamK from Mcas is independent of metal ions and nucleotides except for ATP, and is assembled into well-ordered filamentous bundles consisted of multiple filaments. Our results suggest a dynamically stable assembly of MamK from the uncultivated Nitrospirae MTB that synthesizes multiple magnetosome chains per cell. These findings further improve the current knowledge of biomineralization and organelle biogenesis in prokaryotic systems.
Actins ; chemistry ; metabolism ; Adenosine Triphosphate ; metabolism ; Bacteria ; classification ; metabolism ; Bacterial Proteins ; chemistry ; metabolism ; Magnetospirillum ; classification ; metabolism ; Nucleotides ; metabolism ; Phylogeny ; Substrate Specificity

Objective The aim of this study is to evaluate the shrinkage mode of the primary tumor in women with breast cancer after neoadjuvant chemotherapy ( NAC ) determined by part?mount sub?serial section ( PMSS) and three?dimensional ( 3D) reconstruction technique. Methods Eighty?six women with pathologically proven solitary invasive ductal carcinoma (ⅡA?ⅢC) were recruited. They were divided into two groups. Group A ( n=25) received half cycles of NAC and Group B ( n=61) received whole cycles of NAC. Breast specimen was prepared with PMSS, and residual tumors were microscopically outlined, scanned and registered by Photoshop software. The 3D model of residual tumors was reconstructed with 3D?Doctor software to evaluate the shrinkage mode. Further, the clinicpathologic shrinkage modes were divided into 2 categories:concentric shrinkage mode ( CSM, the longest diameter of the pathological residual tumors was less than 50% and ≤2 cm in comparison with the primary tumor before NAC ) , and non?concentric shrinkage mode ( NCSM, the longest diameter of the pathological residual tumors was more than 50% and/or>2 cm in comparison with the primary tumor before NAC) . Results Pathological shrinkage modes:Group A: modes Ⅰ,Ⅱ, andⅤwere observed in 1, 1, and 23 cases, respectively;Group B:modesⅠ,Ⅱ,Ⅲ,Ⅳ, and Ⅴwere observed in 18, 3, 12, 21, and 7 cases, respectively ( P<0.001) . The multivariate analysis showed that patients with lower primary tumor stage, PR(-) or mammographic malignant calcification before NAC(-) and lymph nodes down?staging after NAC were more likely to present with CSM after NAC ( P<0.05 for all). Conclusions The pathologic reconstruction of breast residual tumors can fully and three?dimensionally reveal the shrinkage mode of the primary breast tumor in women with breast cancer after NAC. PMSS and 3D reconstruction of pathology provide a new platform in this area. Primary tumor stage, PR expression and mammographic malignant calcification before NAC and lymph node down?staging after NAC are independent predictors of the clinicopathologic shrinkage mode.

Objective The aim of this study is to evaluate the shrinkage mode of the primary tumor in women with breast cancer after neoadjuvant chemotherapy ( NAC ) determined by part?mount sub?serial section ( PMSS) and three?dimensional ( 3D) reconstruction technique. Methods Eighty?six women with pathologically proven solitary invasive ductal carcinoma (ⅡA?ⅢC) were recruited. They were divided into two groups. Group A ( n=25) received half cycles of NAC and Group B ( n=61) received whole cycles of NAC. Breast specimen was prepared with PMSS, and residual tumors were microscopically outlined, scanned and registered by Photoshop software. The 3D model of residual tumors was reconstructed with 3D?Doctor software to evaluate the shrinkage mode. Further, the clinicpathologic shrinkage modes were divided into 2 categories:concentric shrinkage mode ( CSM, the longest diameter of the pathological residual tumors was less than 50% and ≤2 cm in comparison with the primary tumor before NAC ) , and non?concentric shrinkage mode ( NCSM, the longest diameter of the pathological residual tumors was more than 50% and/or>2 cm in comparison with the primary tumor before NAC) . Results Pathological shrinkage modes:Group A: modes Ⅰ,Ⅱ, andⅤwere observed in 1, 1, and 23 cases, respectively;Group B:modesⅠ,Ⅱ,Ⅲ,Ⅳ, and Ⅴwere observed in 18, 3, 12, 21, and 7 cases, respectively ( P<0.001) . The multivariate analysis showed that patients with lower primary tumor stage, PR(-) or mammographic malignant calcification before NAC(-) and lymph nodes down?staging after NAC were more likely to present with CSM after NAC ( P<0.05 for all). Conclusions The pathologic reconstruction of breast residual tumors can fully and three?dimensionally reveal the shrinkage mode of the primary breast tumor in women with breast cancer after NAC. PMSS and 3D reconstruction of pathology provide a new platform in this area. Primary tumor stage, PR expression and mammographic malignant calcification before NAC and lymph node down?staging after NAC are independent predictors of the clinicopathologic shrinkage mode.

Objective:This study was conducted to evaluate the roles of internal mammary sentinel lymph node biopsy (IM-SL-NB) in the treatment of breast cancer patients with clinically positive axillary lymph nodes. Methods:This study is a one-armed clini-cal research conducted from June 2013 to October 2014. A total of 64 breast cancer patients from Shandong Cancer Hospital with clini-cally positive axillary lymph nodes were enrolled in the study. All patients underwent axillary lymph node dissection. Meanwhile, IM-SLNB was performed in all patients using the new injection method of radiotracer. Results:Among the 64 enrolled patients, the visual-ization rate of internal mammary lymph node was 59.4%(38/64). For the 38 patients who were subjected to visualization of the internal mammary node, the detection rate was 100%(38/38), and the incidence of complications was 7.9%(3/38). The metastasis rate of inter-nal mammary lymph node was 21.1%(8/38). Patients with upper inner quadrant tumors and metastasis of more axillary lymph nodes had a significantly higher chance of developing sentinel lymph node metastasis (P<0.001 and P=0.017, respectively) than the other pa-tients. The clinical benefit rate of the above mentioned treatment was 59.4%. Among the patients, 12.5%(8/64) received extra internal mammary radiotherapy, whereas 46.9%(30/64) patients avoided the unnecessary internal mammary radiotherapy. Conclusion:IM-SL-NB should be performed in breast cancer patients with clinically positive axillary lymph nodes because IM-SLNB could provide the ac-curate indication of radiation to the internal mammary area, especially for the patients with upper inner quadrant tumors and those with a suspiciously high level of axillary lymph node metastasis.