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.

This paper briefly introduces the working procedure of in vitro diagnostic products (IVD) industrial standards, and elaborates the importance of professional standards for production and supervision. Based on the analysis of working progress during the past 10 years, some problems and countermeasures on project setting, participation, standard material, personnel training, work cycle are put forward, which are helpful for the future development of the IVD.
Diagnostic Techniques and Procedures ; standards ; Humans ; Reference Standards

Objective To explore the clinical variables associated with the shrinkage modes of primary breast tumor in women after neoadj uvant chemotherapy (NAC ), and to develop a nomogram for predicting non-concentric shrinkage mode(NCSM).Methods Sixty-one women with pathologically proven solitary invasive ductal carcinoma (ⅡA-ⅢC)were recruited. Breast specimen was prepared with PMSS, and residual tumors were microscopically outlined,scanned and registered by Photoshop CS 5 software.The 3D model of residual tumors was reconstructed with 3D-DOCTOR 4.0 software to evaluate the shrinkage mode.17 factors such as age and body mass index and menopausal status were chosen as independent variables,and the clinic-pathologic shrinkage mode was considered as dependent variable. A Logistic regression model was used to construct the nomogram. Results Primary tumor stage,lymph node down-staging, PR and mammographic malignant calcification before NAC were independent predictors of clinic-pathologic shrinkage mode (β:1.538,OR:4.656,95%CI:1.414-15.328,P=0.011;β:1.555,OR:4.735, 95%CI:1.082-20.722,P=0.039;β:-1.707, OR:0.181, 95%CI:0.044-0.741,P = 0.017;β:- 1.405, OR:3.808, 95% CI:0.06 - 0.998,P = 0.048, respectively ). The nomogram predicting the risk of NCSM showed a good concordance index(0.869),and its conformity of mean absolute error was 0.039. Conclusion Based on the clinicopathological findings of primary breast tumor, a nomogram to predict shrinkage modes after NAC in breast carcinoma patients is constructed.The statistical tool is helpful for individually selecting the patients who can be treated with BCT after NAC.

Objective To introduce the clinical application of the extended anterolateral leg flap,which is based on the superficial peroneal artery and lateral supramalleolar artery. Methods Through anatomic study on the blood supplies of the skin over the anterolateral portion of the leg.The combined superficial peroneal artery and lateral supramalleolar artery could extend the size of the traditional anterolateral leg flap.In addition,the point of pivot of the extended anterolateral leg flap could locate at the level of the lateral malleolus,which decreases the distance between the flap and recipient site.Clinically,we have used the extended anterolateral leg flaps in 60 patients to cover skin defect over the feet.The flap ranged from 16 cm×10 cm(pedicle length 8.0 cm)to 26 cm×7 cm(pedicle length 6.0 cm). Results In the series, the flap survived in 53 patients(88.3%)unevently,and partial necrosis occurred in 7 patients(11.7%).Conclusion Based on the combined superficial peroneal artery and lateral supramalleolar artery,the size of the traditional anterolateral leg flap could be inceased,which enlarges its value on the coverage of skin defects over the feet.

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.

Objective To verify the accuracy of the modified technique for internal mammary sentinel lymph node biopsy ( IM-SLNB) in breast cancer .Methods In the validation study , the radiotracer was injected with the modified technique , and fluorescence tracer was injected into the peritumoral breast tissue .The radioac-tive IM-SLN was identified by preoperative lymphoscintigraphy and γprobe.The radioactive IM-SLN received bi-opsy during operation .The status of the fluorescence tracer was identified by the fluorescence imaging system . Results A total of 162 patients were enrolled from Sep .2013 to Dec.2014.IM-SLNB was performed in 110 pa-tients.The radiotracer and the fluorescence tracer were identified in the same IM-SLN in 94 cases, and the con-cordance rate was 85.5%(Case-base, Spearman coefficient correlation 0.823, P<0.001).Conclusion Dif-ferent tracers injected into the different sites of the intra-parenchyma can reach the same IM-SLN, proving the ac-curacy of the modified technique and the hypothesis of IM-SLN lymphatic drainage pattern ( IM-SLN receives not only the lymphatic drainage from the primary tumor area but the entire breast parenchyma ) .

ObjectiveTo evaluate the risk factors for sentinel lymph node(SLN)metastasis,and assess the value of the Memorial Sloan Kettering Cancer Center (MSKCC) nomogram in predicting SLN metastasis in breast cancer patients.Methods A sentinel lymph node biopsy(SLNB) database containing 1227 consecutive breast cancer patients(412 patients with a positive SLN)was retrospectively analyzed.The value of MSKCC nomogram was predicted by drawing the trend line and calculating the area under the curve(AUC) of receiver-operator characteristic(ROC). Meanwhile,the risk factors of SLN metastasis were evaluated. Results Tumor size,tumor location,histological grade,lymphovascular invasion,mulifocality,ER and PR status were correlated with SLN metastasis( all P ＜ 0.05 ).All the above factors but tumor location were significant independent predictors for SLN metastasis(all P ＜ 0.01 ).The MSKCC nomogram presented AUC value of 0.730 for ROC.Patients with predictive values lower than 16％ had the frequency of 0.9％ for SLN metastasis while patients with predictive values higher than 70％ had the frequency of 96.2％.ConclusionsThe risk factors of SLN metastasis in our study are consistent with those in MSKCC nomogram.MSKCC nomogram is a useful tool in predicting the probability of SLN metastasis for breast cancer patients.Axillary surgery can be avoided in patients with the predictive values lower than 16％,axillary lymph node dissection could be done in patients with the predictive values higher than 70％,and other patients should still undergo SLNB.

OBJECTIVETo study the effects of polypeptide extract from scorpion venom (PESV) alliance with chemotherapy on angiogenesis of Lewis lung carcinomas (LLC) and its mechanism.

METHODLLC cells suspension (4 x 10(6) cells/mL) were subcutaneously injected into 54 C57BL/6J mice in right armpits. Then the tumor-bearing mice were randomly divided into three groups: the control group, the chemotherapy group and the PESV group. Cyclophosphamide was used to establish the model of cancer. Chemotherapy and PESV were added to the PESV group. Every 7 days, 6 mice of each group were executed, and the experiments were carried out for 28 days. The tumor volume and inhibitory rate were determined. Immunohistochemistry and RT-PCR were used to determine the expression of factor VIII, alpha-SMA, Dll4 and Notch1 in tumor tissue. Correlation analysis was used to identify the relationship of factor VIII and calculate microvessel density (MVD), alpha-SMA and vascular maturity.

RESULTThe inhibitory rate of PESV was 42.21%. Comparing with the chemotherapy group, the expression of tumor factor Dll4 and Notch1 in the PESV group were decreased significantly (P < 0.05). The expression of factor VIII and alpha-SMA in the chemotherapy group is lower than the control group (P < 0.05), while it's higher when compared with the PESV group (P < 0.01). Expression of Dll4 and Notch1 in the chemotherapy group at the 28th day were higher than the control group (P < 0.05), and the expression in the PESV group at the 21st day were significantly lower than the chemotherapy group (P < 0.05).

CONCLUSIONPESV could inhibit the angiogenesis of LLC. It might be attained by decreasing the level of angiogenic factors, that are factor VIII, alpha-SMA, Dll4 and Notch1 in tumor microenvironment.

Animals ; Antineoplastic Agents ; blood ; chemistry ; therapeutic use ; Carcinoma, Lewis Lung ; drug therapy ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; drug therapy ; Peptides ; chemistry ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction ; Scorpion Venoms ; chemistry

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 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.