Objective CD44, a cell surface glycoprotein, plays an important role in tumor growth and glycolysis.The aim of this study was to investigate the effects of neutralizing CD44 antibodies on the growth and glycolytic metabolism of B16 cells in melanoma in vitro.Methods B16 cells were treated with control antibodies (50 μg/mL) or different concentrations of CD44 antibodies (2, 10, and 50 μg/mL) for 24 hours, followed by examination of the activation of the AKT pathway in the B16 cells by Western blot.Then the tumor cells were also treated with control antibodies (50 μg/mL) or CD44 antibodies (50μg/mL) after pretreated with API-2 (4 μmol/L) in a parallel test.After 48 hours of treatment, the expression of lactate dehydrogenase A (LDHA) in the B16 cells and the level of lactate in the culture supernatant were detected by immunofluorescence and colorimetry, respectively.Lastly, the B16 cells were treated with control antibodies (50μg/mL), API-2 (4 μmol/L), CD44 antibodies (50μg/mL), or API-2 + CD44 antibodies for 96 hours, followed by measurement of the proliferation of the cells by MTT and their apoptosis by AO/EB and AnnexinV staining.Results In comparison with the control antibody group, the level of AKT phosphorylation (p-AKT) in the B16 cells showed a concentration-dependent increase in the 2, 10, and 50 μg/mL CD44 antibody groups (1.00±0.25 vs 2.51±0.32, 3.89±0.46, and 4.07±0.42, P<0.01), and the expression of LDHA was increased by (2.13±0.24) times, with the lactate level in the culture supernatant significantly elevated from (35.32±3.24) to (56.34±8.19) mmol/L (P<0.01) after 96 hours of treatment with 50 μg/mL CD44 antibodies.Treatment with API-2+CD44 antibodies, however, suppressed the increase in the LDHA expression and reduced the level of lactate.Compared with the control antibody group, the proliferation rate of the B16 cells was markedly decreased in the API-2, CD44 antibody, and API-2+CD44 antibody groups ([103±12.91] vs [84.87±19.35], [71.35±16.23], and [41.16±9.15]%, P<0.05), while the apoptosis rate remarkably increased ([5.23±0.96] vs [13.65±4.27], [19.21±3.53], and [43.21±7.87]%, P<0.01).Conclusion Neutralizing the function of CD44 in the B16 cells in vitro can inhibit the growth of the cells and promote AKT-mediated glycolytic metabolism, while suppressing the AKT pathway may enhance the antitumor activity of the CD44 antibody.

Objective To observed the expression of serum TNF-α and IL-10 in rats with severe acute pancreatitis (SAP) at different altitudes,and to explore the relationships between TNF-α and IL-10,the pathological changes of the pancreas,and the experimental basis for clinical diagnosis and treatment of SAP.Methods 72 specific pathogen free (SPF) Wistar male rats were divided randomly into three groups:1 500 meters altitude (group L),3 300 meters altitude (group M),and 4 300 meters altitude (group H).These three groups were then each divided randomly into four subgroups:control (group n),6 hours after pancreatitis (group p 6 h),12 hours after pancreatitis (group p 12 h),and 24 hours after pancreatitis (group p 24 h).Pancreatitis was induced by intraductal administration of 5％ sodium taurocholate hydrate (NaTc).The rats were killed at 6,12,and 24 hours after NaTc injection in groups p.The group n rats were killed after 6 hours of pancreas observation.Blood samples and pancreatic tissues were collected post mortem and enzyme-linked immunosorbent assay (ELISA) measured serum TNF-α and IL-10.Results Compared with the control (group n),histopathological scores,IL-10,and TNF-α in the same altitude had a significant difference (P ＜ 0.05) in group p at each time point.In the same altitude of group p,histopathological scores and IL-10 were increased with time elapsed (P ＜ 0.05),while TNF-α was decreased with time elapsed (P ＜ 0.05).There was a significant difference between group Mp and Lp in histopathological scores,IL-10,and TNF-α (P ＜ 0.05),and the same result between group Hp and Lp (P ＜ 0.05),but there was no significant difference between group Hp and Mp (P ＜ 0.05).Meanwhile,IL-10 had a positive relationship with histopathological score,but TNF-α had a negative relationship with histopathological score.Conclusions The level of TNF-α increased with increasing altitude but significantly reduced with elapsed time.The level of IL-10 increased with both increasing altitude elapsed time.These results suggested that TNF-o and IL-10 might play a important role at different times in severe acute pancreatitis.

Objective To analyze the prognostic factors and their influences on breast cancer patients with local-regional recurrence.Methods From Jan.1st,1998 to Dec.30th,2007,66 breast cancer patients with local-regional recurrence were treated at Breast Center of Peking University People's Hospital.The overall survival of the patients was analyzed using Kaplan Meier survival analysis and the prognostic factors such as primary tumor status,initial treatment,location of recurrence and treatment strategy after local recurrence were analyzed using Cox regression model.Results The median follow-up was 111 months.The median survival for the 66 patients with recurrence was 99 months,ranging from 3 to 188 months and 5-year survival rate was 60％.Kaplan-Meier analysis revealed statistically significant better survival for patients with unifocal primary tumor,undergoing initial breast conservation surgery,with local recurrence only and undergoing radical local treatment after local recurrence (P =0.003,P =0.017,P =0.050,and P =0.000 respectively).COX regression analysis showed that age at initial diagnosis and radical local treatment after recurrence had influences on post-recurrence survival of patients without distant metastasis (P =O.004 and 0.000 respectively).Conclusion The clinical and pathological features of primary tumor,initial surgery,site of recurrence and treatment strategy after recurrence have influences on prognosis of breast cancer patients with local-regional recurrence.

Objective:To evaluate the frequency and predictive factors of nipple involvement in a large contemporary cohort of pa-tients and to improve patient selection for the preservation of the nipple-areolar complex. Methods: This retrospective study re-viewed the medical charts of 1,190 patients who underwent traditional mastectomy in Peking University People's Hospital between October 2008 and March 2014. Nipple involvement incidence was compared between the cases of clinically abnormal and clinically normal breasts. Other clinicopathological features and nipple status were analyzed to evaluate the association between these factors and occult nipple involvement. Univariate and multivariate analyses were conducted to identify predictive factors. Results:Nipple in-volvement was detected in 6.0%of the mastectomy specimens. Meanwhile, incidence was 40.7%(22 out of 54) in clinically abnormal nipple cases and 4.3%(49 out of 1,136) in clinically normal nipple cases (χ2=121.9, P<0.001). Univariate analysis revealed that tumor lo-cation, tumor to nipple distance (≤2 and>2 cm), lymphovascular invasion, diameter (including carcinoma in situ;≤3.5 and>3.5 cm), T stage, N stage, and TNM stage were associated with occult nipple involvement. By logistic regression analysis, tumor location, tumor to nipple distance, T stage, and N stage were determined to be the independent predictors of nipple involvement. Conclusion:Clinical abnormalities of the nipples are reliable and potent predictors of nipple pathology. The cases with peripheral tumor, T1-T2 stage, and N0-N1 stage have lower probability of occult nipple involvement.

Objective To evaluate the long-term safety of sentinel lymph node biopsy mapped by combination of indocyanine green and methylene blue in breast cancer patients.Methods 198 breast cancer patients with clinical negative axillary lymph node received sentinel lymph node biopsy mapped by combination of indocyanine green and methylene blue.Patients were followed up and regional lymph node recurrence,disease free survival(DFS) and overall survival(OS) were analyzed.Results After a median follow-up of 70 months,2 patients had ipsilateral lymph node recurrence with a regional lymph node recurrence rate of 1％ (2/198).14 patient had recurrence or metastasis and 6 patients died of distant metastasis.The estimated 6 years DFS was 94.4％ and OS was 96.5％.The incidence of arm lymphoedema within patients who received axillary lymph node dissection was 4.5％ and it was 2.5％ in patients who received sentinel lymph node biopsy.Conclusions The sentinel lymph node biopsy mapped by combination of indocyanine green and methylene blue was safe and reliable method for further staging axillary lymph node stastus.

Objective:To examine treatment outcomes of breast phyllodes tumors and the prognosis factors of local recurrence.Methods:This retrospective cohort study included 276 patients who underwent surgical resection at Breast Center, Peking University People′s Hospital from January 2011 to December 2019. Tumor subtype and histopathological features were determined from pathology reports, and the deadline of follow-up was September 30 th, 2020. All 276 patients underwent open surgery, including 17 patients of mastectomy, and 259 patients of lumpectomy. The enrolled patients were all female, with age of (41.5±11.3) years (rang: 11 to 76 years), and tumor diameter of 35(28) mm ( M( Q R)). The Kaplan-Meier method and Log-rank test were used for survival analysis. The multivariate analysis was implemented using the Cox proportional hazard model. Results:According the pathologic test, there were 191 patients of benign phyllodes tumor, 67 patients of borderline tumor and 18 patients of malignant tumor. There were 249 patients with a follow-up of more than 6 months, and 14.1% (35/249) had local recurrence. The time-to-recurrence was (28.6±22.2) months (range: 2 to 96 months), (29.1±18.1) months (range: 2 to 80 months), (32.1±30.1) months (range: 5 to 96 months) and (12.0±6.9) months (range: 8 to 20 months) for benign, borderline and malignant phyllodes tumors. Tumor diameter (≥100 mm vs.<50 mm, HR=3.968, 95%CI: 1.550 to 10.158, P=0.004) and malignant heterologous element (yes vs. no, HR=26.933, 95%CI: 3.105 to 233.600, P=0.003) were prognosis factors of local recurrence. One death from malignant phyllodes occurred after distant metastasis. The 3-year disease-free survival rates of benign, borderline and malignant phyllodes tumor were 88.2%, 81.7% and 81.4% ( P=0.300). Conclusion:Phyllodes tumors have a considerable local recurrence rate, which may be associated with tumor diameter and malignant heterologous element.

Objective:To examine treatment outcomes of breast phyllodes tumors and the prognosis factors of local recurrence.Methods:This retrospective cohort study included 276 patients who underwent surgical resection at Breast Center, Peking University People′s Hospital from January 2011 to December 2019. Tumor subtype and histopathological features were determined from pathology reports, and the deadline of follow-up was September 30 th, 2020. All 276 patients underwent open surgery, including 17 patients of mastectomy, and 259 patients of lumpectomy. The enrolled patients were all female, with age of (41.5±11.3) years (rang: 11 to 76 years), and tumor diameter of 35(28) mm ( M( Q R)). The Kaplan-Meier method and Log-rank test were used for survival analysis. The multivariate analysis was implemented using the Cox proportional hazard model. Results:According the pathologic test, there were 191 patients of benign phyllodes tumor, 67 patients of borderline tumor and 18 patients of malignant tumor. There were 249 patients with a follow-up of more than 6 months, and 14.1% (35/249) had local recurrence. The time-to-recurrence was (28.6±22.2) months (range: 2 to 96 months), (29.1±18.1) months (range: 2 to 80 months), (32.1±30.1) months (range: 5 to 96 months) and (12.0±6.9) months (range: 8 to 20 months) for benign, borderline and malignant phyllodes tumors. Tumor diameter (≥100 mm vs.<50 mm, HR=3.968, 95%CI: 1.550 to 10.158, P=0.004) and malignant heterologous element (yes vs. no, HR=26.933, 95%CI: 3.105 to 233.600, P=0.003) were prognosis factors of local recurrence. One death from malignant phyllodes occurred after distant metastasis. The 3-year disease-free survival rates of benign, borderline and malignant phyllodes tumor were 88.2%, 81.7% and 81.4% ( P=0.300). Conclusion:Phyllodes tumors have a considerable local recurrence rate, which may be associated with tumor diameter and malignant heterologous element.

Objective:To investigate the application effect of Press-Z-track-Turn (PZT) intramuscular injection on the extravasation, local reaction, local pain after injection and patient satisfaction after Fulvestrant injection.Methods:Used a self-controlled study, 63 patients who underwent bilateral intramuscular injection of Fluvastatin in the outpatient injection room of Tianjin Medical University Cancer Hospital from August to October 2022 were conveniently selected. The observation time was two treatment cycles after inclusion in this study. Before the injection of the first treatment cycle, a coin toss was used to randomly decide to use traditional intramuscular injection and PZT intramuscular injection method for injection and record. After 28 days, another injection method was used, with a total of 126 injections per injection method. Observed the extravasation of drug solution and local reactions at the injection site within 7 days after injection using two injection methods, as well as the pain and satisfaction of patients within 7 days after injection.Results:The extravasation rate of drug solution for the PZT intramuscular injection was 1.59% (2/126), which was lower than 7.14% (9/126) for the traditional intramuscular injection, and the difference was statistically significant ( χ2=4.66, P<0.05); the incidence of mild and moderate local reactions for the PZT intramuscular injection was 3.97% (5/126) and 1.59% (2/126), respectively, lower than that for the traditional intramuscular injection 11.11% (14/126) and 5.56% (7/126), and the difference was statistically significant ( χ2=8.26, P<0.05); the pain score of the PZT intramuscular injection was (2.08 ± 0.85) points, lower than that of the traditional intramuscular injection (5.03 ± 1.06) points, and the satisfaction score of the PZT intramuscular injection was (23.68 ± 1.64) points, higher than that of the traditional intramuscular injection (20.10 ± 2.58) points. The difference was statistically significant ( t=-17.24, 9.16, both P<0.01). Conclusions:PZT intramuscular injection can effectively reduce the overflow rate, the incidence of grade 1 and 2 local reactions and the degree of pain of patients, and improve patient satisfaction, which is worthy of clinical application.