Photothermal-Controllable Microneedles with Antitumor, Antioxidant, Angiogenic, and Chondrogenic Activities to Sequential Eliminate Tracheal Neoplasm and Reconstruct Tracheal Cartilage.

The optimal treatment for tracheal tumors necessitates sequential tumor elimination and tracheal cartilage reconstruction. This study introduces an innovative inorganic nanosheet, MnO2 /PDA@Cu, comprising manganese dioxide (MnO2 ) loaded with copper ions (Cu) through in situ polymerization using polydopamine (PDA) as an intermediary. Additionally, a specialized methacrylic anhydride modified decellularized cartilage matrix (MDC) hydrogel with chondrogenic effects is developed by modifying a decellularized cartilage matrix with methacrylic anhydride. The MnO2 /PDA@Cu nanosheet is encapsulated within MDC-derived microneedles, creating a photothermal-controllable MnO2 /PDA@Cu-MDC microneedle. Effectiveness evaluation involved deep insertion of the MnO2 /PDA@Cu-MDC microneedle into tracheal orthotopic tumor in a murine model. Under 808 nm near-infrared irradiation, facilitated by PDA, the microneedle exhibited rapid overheating, efficiently eliminating tumors. PDA's photothermal effects triggered controlled MnO2 and Cu release. The MnO2 nanosheet acted as a potent inorganic nanoenzyme, scavenging reactive oxygen species for an antioxidant effect, while Cu facilitated angiogenesis. This intervention enhanced blood supply at the tumor excision site, promoting stem cell enrichment and nutrient provision. The MDC hydrogel played a pivotal role in creating a chondrogenic niche, fostering stem cells to secrete cartilaginous matrix. In conclusion, the MnO2 /PDA@Cu-MDC microneedle is a versatile platform with photothermal control, sequentially combining antitumor, antioxidant, pro-angiogenic, and chondrogenic activities to orchestrate precise tracheal tumor eradication and cartilage regeneration.

Low-dose carboplatin modifies the tumor microenvironment to augment CAR T cell efficacy in human prostate cancer models.

Chimeric antigen receptor (CAR) T cells have transformed the treatment landscape for hematological malignancies. However, CAR T cells are less efficient against solid tumors, largely due to poor infiltration resulting from the immunosuppressive nature of the tumor microenvironment (TME). Here, we assessed the efficacy of Lewis Y antigen (LeY)-specific CAR T cells in patient-derived xenograft (PDX) models of prostate cancer. In vitro, LeY CAR T cells directly killed organoids derived from androgen receptor (AR)-positive or AR-null PDXs. In vivo, although LeY CAR T cells alone did not reduce tumor growth, a single prior dose of carboplatin reduced tumor burden. Carboplatin had a pro-inflammatory effect on the TME that facilitated early and durable CAR T cell infiltration, including an altered cancer-associated fibroblast phenotype, enhanced extracellular matrix degradation and re-oriented M1 macrophage differentiation. In a PDX less sensitive to carboplatin, CAR T cell infiltration was dampened; however, a reduction in tumor burden was still observed with increased T cell activation. These findings indicate that carboplatin improves the efficacy of CAR T cell treatment, with the extent of the response dependent on changes induced within the TME.

[Analysis of the efficacy and safety of CT-guided radiofrequency ablation of posterior root of the spinal nerve in the treatment of postherpetic neuralgia].

Objective: To investigate the efficacy and safety of CT-guided radiofrequency ablation of posterior root of spinal nerve in the treatment of postherpetic neuralgia (PHN). Methods: A total of 102 PHN patients (42 males and 60 females) aged (69.7±9.4) years who underwent CT-guided radiofrequency ablation of posterior root of spinal nerve in the Department of Pain Medicine of the Affiliated Hospital of Jiaxing University from January 2017 to April 2020 were retrospectively included. Patients were followed up, and numerical rating scale (NRS) score, Pittsburgh sleep quality index (PSQI), satisfaction score and complications before surgery (T0) and at 1 d (T1), 3 months (T2), 6 months (T3), 9 months (T4) and 12 months (T5) after surgery were recorded. Results: The NRS score of PHN patients at T0, T1, T2, T3, T4, and T5 [M(Q1, Q3)] was 6(6, 7), 2(2, 3), 3(2, 4), 3(2, 4), 2(1, 4), 2(1, 4), respectively. Likewise, the PSQI score [M(Q1, Q3)] at aforementioned time points was 14(13, 16), 4(3, 6), 6(4, 8), 5(4, 6), 4(2, 8), 4(2, 9), respectively. Compared with T0, the NRS and PSQI scores at all time points from T1 to T5 were lower, with statistically significant differences (all P<0.001). The overall effective rate of surgery at 1 year postoperatively was 71.6% (73/102) with a satisfaction score of 8(5, 9), and the recurrence rate was 14.7% (15/102) with a recurrence time of (7.5±0.8) months. The main postoperative complication was numbness, with an incidence of 86.0% (88/102), and the degree of numbness gradually decreased with time. Conclusion: CT-guided radiofrequency ablation of posterior root of spinal nerve for PHN has a high effective rate and a low recurrence rate, with high safety profile, and may be a feasible surgical option for the treatment of PHN.

[The correlation between methylation in HPV16 long control region and cervical intraepithelial neoplasia grade 2 or more: a Meta-analysis].

Objective: To investigate the correlation between methylation in human papillomavirus 16 (HPV16) long control region (LCR) and cervical intraepithelial neoplasia grade ≥2 (CIN2+). Methods: The literature retrieval was conducted by using the databases of PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang data and Weipu according to the inclusion and exclusion criteria, and the retrieval period was from the establishment of the databases to February 27th, 2022. Software RevMan 5.3 and Stata 15.1 were used for Meta-analysis. Results: A total of 17 literatures were included involving 1 421 subjects. Results of Meta-analysis showed that OR of the correlation between methylation of HPV16 LCR and CIN2+ was 1.56 (95%CI: 0.70-3.47). Subgroup analysis showed that methylation of the 5' terminal, enhancer and promoter regions were not associated with CIN2+, while in four E2 binding sites (E2BS), the methylation of E2BS1, E2BS3 and E2BS4 increased the risk of CIN2+, with the ORs of 3.92 (95%CI: 1.92-7.99), 10.50 (95%CI: 3.67-30.04) and 3.65 (95%CI: 1.58-8.41), respectively. However, subgroup analysis on E2BS2 was not performed due to the limitation of the number of literatures. According to the different sources of population, the risk of CIN2+ in Chinese population was associated with methylation of HPV16 LCR (OR=2.14, 95%CI: 1.31-3.50). There was a correlation between the risk of CIN2+ and HPV16 LCR methylation in the population with pyrosequencing of HPV16 LCR, and OR was 1.75 (95%CI: 1.03-2.98). Conclusion: The risk of CIN2+ is correlated with the methylation of E2BS in HPV16 LCR, which can be used as potential biomarkers.

[The role of STAT-6/KLF-4/PPAR-γ activation in alveolar macrophage polarization changes in silica-induced pulmonary fibrosis].

Objective: To observe the effect of silicon dioxide (SiO(2)) on the polarization of alveolar macrophages (AMs) , and to explore the expressions and the significance of signal transducer and activator of transcription-6 (STAT-6) /Krüppel-like factor-4 (KLF-4) /peroxisome proliferators-activated receptors-γ (PPAR-γ) signaling molecules in AMs. Methods: In November 2020, C57BL/6 mice were randomly divided into crystalline SiO(2) group and normal saline (NS) group, and 12 mice in each group. Mice were intratracheally instillated with 100 µl crystalline SiO(2) suspension (20 mg/ml) or 100 µl NS, and were sacrificed after 28 days. Masson staining was used to observe the degree of pulmonary fibrosis of mice and hydroxyproline (HYP) level were assessed. The proportions of M1-typed and M2-typed AMs in bronchoalveolar lavage fluid (BLAF) were analyzed by flow cytometry. The mRNA relative expression levels of inducible nitric oxide synthase (iNOS) , arginidase-1 (Arg-1) , interleukin (IL) -1ß, tumor necrosis factor-α (TNF-α) , IL-6, IL-10, transforming growth factor-ß (TGF-ß) , STAT-6, KLF-4 and PPAR-γ were detected by real-time fluorescence quantitative PCR. Activities of iNOS and Arg-1, as well as contents of IL-1ß, TNF-α, IL-6, IL-10 and TGF-ß were assessed by the enzyme-linked immunosorbent. The protein relative expression levels of phosphorylation-signal transducer and activator of transcription-6 (p-STAT-6) , KLF-4 and PPAR-γ were evaluated by immunofluorescence. Results: After 28 days of treatment, the structure of the lung tissue of the mice was destroyed, and the deposition of collagen was significantly increased in the crystalline SiO(2) group. Compared with NS group, HYP level of lung tissue in crystalline SiO(2) group were increased, the proportion of M2-typed AMs in crystalline SiO(2) group was increased, the proportion of M1-typed AMs in crystalline SiO(2) group was decreased, the mRNA relative expressions and contents of Arg-1, IL-10, TGF-ß in crystalline SiO(2) group were significantly increased, the mRNA relative expressions and contents of iNOS, IL-1ß, TNF-α, IL-6 in crystalline SiO(2) group were significantly decreased, the mRNA of STAT-6, KLF-4, PPAR-γ and the protein relative expression levels of p-STAT-6, KLF-4, PPAR-γ were significantly increased in crystalline SiO(2) group, and the the differences were statistically significant (P<0.05) . Conclusion: Crystalline SiO(2) may mediate the process of pulmonary fibrosis through promote AMs polarization toward M2-typed by activating the STAT-6/KLF-4/PPAR-γ signaling pathway.

[A nested case-control study on the relationship between red blood cell folate and the prognosis of low-grade cervical intraepithelial neoplasia].

Objective: To evaluate the relationship between red blood cell folate (RBC folate) and the prognosis of low-grade cervical intraepithelial neoplasia (CIN 1). Methods: In the married women cohort established in 2014, 564 women with CIN 1 diagnosed by pathology were recruited. The demographic characteristics and factors of cervical intraepithelial neoplasia were collected. Meanwhile, the infection status of human papillomavirus (HPV) was detected by molecular diversion hybridization, and the level of RBC folate was measured by chemical photoimmunoassay. After 24 months of follow-up, pathological examination was performed again to observe the prognosis of participants. The women with reversal were taken as the control group,and those with continuous and progressive CIN 1 were taken as the case group respectively. The relationship between RBC folate and CIN 1 outcome was evaluated by logistic regression model. Results: 453 women completed the follow-up, aged (49.72±6.84) years old. CIN 1 was reversed in 342 women, continued in 58 cases and progressed in 53 cases. The RBC folate level M (Q1,Q3) were 399.01 (307.10, 538.97) ng/ml, 316.98 (184.74, 428.49) ng/ml and 247.14 (170.54, 348.97) ng/ml, respectively. With the decrease of RBC folate, the risk of continuous and progressive CIN 1 increased (all P<0.001), while the risk of reversal CIN 1 decreased gradually (P<0.001). Combined with high-risk human papillomavirus (HR-HPV) infection status, low level of RBC folate could increase the risk of CIN 1 progression regardless of HR-HPV infection (HR-HPV infection: OR=21.34, 95%CI: 3.98-114.54; HR-HPV uninfection: OR=11.15, 95%CI: 2.34-53.13). Conclusion: Low level of RBC folate could increase the risk of CIN 1 persistence and progression regardless of HR-HPV infection.

Early-phenotype CAR-T cells for the treatment of pediatric cancers.

Chimeric antigen receptor (CAR)-T-cell therapy is a promising approach for the treatment of childhood cancers, particularly high-risk tumors that fail to respond to standard therapies. CAR-T cells have been highly successful in treating some types of hematological malignancies. However, CAR-T cells targeting solid cancers have had limited success so far for multiple reasons, including their poor long-term persistence and proliferation. Evidence is emerging to show that maintaining CAR-T cells in an early, less-differentiated state in vitro results in superior persistence, proliferation, and antitumor effects in vivo. Children are ideal candidates for receiving less-differentiated CAR-T cells, because their peripheral T-cell pool primarily comprises naïve cells that could readily be harvested in large numbers to generate early-phenotype CAR-T cells. Although several studies have reported different approaches to successfully generate early CAR-T cells, there are only a few clinical trials testing these in adult patients. No trials are currently testing early CAR-T cells in children. Here, we summarize the different strategies used to maintain CAR-T cells in an early phenotypic stage and present evidence suggesting that this approach may be particularly relevant to treating childhood cancers.

MiR-101 promotes nasopharyngeal carcinoma cell apoptosis through inhibiting Ras/Raf/MEK/ERK signaling pathway.

The article "MiR-101 promotes nasopharyngeal carcinoma cell apoptosis through inhibiting Ras/Raf/MEK/ERK signaling pathway, by R.-S. Wu, E.-H. Qiu, J.-J. Zhu, J.-R. Wang, H.-L. Lin, published in Eur Rev Med Pharmacol Sci 2018; 22(1): 150-157-DOI: 10.26355/eurrev_201801_14112-PMID: 29364482" has been withdrawn from the authors. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/14112.

[The possible mechanisms of simvastatin on apoptosis of lung adenocarcinoma cells].

Objective: To explore the possible mechanisms of simvastatin-induced apoptosis in lung adenocarcinoma cells. Methods: The experiment was divided into control group (vehicle treated A549 cells), different concentrations (10, 20, 40, 80 mg/L) simvastatin group (simvastatin treated with different concentrations of A549 cells), aspartate specific proteinase (caspase) inhibitor (Z-VAD-FMK) group (50 µmol/L Z-VAD-FMK treated A549 cells), 40 mg/L simvastatin combined with Z-VAD-FMK group (40 mg/L simvastatin combined with 50 µmol/L Z-VAD-FMK co-treated A549 cells), interleukin-6 (IL-6) group (IL-6 acts on A549 cells) and different concentrations (10, 20, 40 mg/L) simvastatin combined with IL-6 group (simvastatin combined with IL-6 act on A549 cells). Cell counting kit-8 (CCK8) method was used to detect the effect on survival rate of lung adenocarcinoma A549 cells; Flow cytometry was used to detect the effect of simvastatin on A549 cell cycle; Mitochondrial membrane potential-1 (JC-1) fluorescent probe was wsed to detect the effect of simvastatin on mitochondrial membrane potential (MMP); Flow-type phosphatidl serine protein antibody Annexin V/propidium iodide (Annexin V-FITC/PI) double staining method was used to detect the effect of simvastatin on A549 cell apoptosis; CCK8 method was used to detect the effect of Z-VAD-FMK on the survival rate of A549 cells; TdT-mediated 2'-deoxyuridine 5'-triphosphate (dUTP) nick end labeling (TUNEL) method was used to detect the effect of Z-VAD-FMK on simvastatin-induced apoptosis in A549 cells; Western blot method was used to detect the effect of simvastatin on the expression levels of Janus kinase 2 and activation of signal transducers and activators of transcription 3 (JAK2/STAT3) pathway-related proteins phosphorylated JAK2 (p-JAK2), JAK2, phosphorylated STAT3 (p-STAT3), and STAT3 before and after the activator IL-6 of JAK2/STAT3 pathway acted on A549 cells. Results: The survival rates of A549 cells in the 20-80 mg/L simvastatin-treated groups were significantly lower than that in the control group (all P<0.05), and gradually decreased with the increase of the concentration of the simvastatin and the extension of the action time. The cells in the G(0)/G(1) phase of the simvastatin group were significantly higher than those in the control group, and the cells in the G(2)/M phase were significantly lower than those in the control group (all P<0.01). The MMP of the treatment group with different concentrations of simvastatin was significantly lower than that of the control group (all P<0.05). The apoptosis rate of the 20 mg/L and 40 mg/L simvastatin-treated group was significantly higher than that of the control group (both P<0.01). The cell survival rate of the 40 mg/L simvastatin group and the 40 mg/L simvastatin combined with Z-VAD-FMK group were (52.2±2.7)% and (57.5±3.8)%, respectively, were lower than that of the control group (100.0±2.7)% (both P<0.01). But the difference between 40 mg/L simvastatin group and the simvastatin combined with Z-VAD-FMK group was not statistically significant (P>0.05). The cell numbers with positive fluorescent staining in the 40 mg/L simvastatin group were significantly more than those in the control group, but the cell numbers with positive fluorescent staining in the 40 mg/L simvastatin combined with Z-VAD-FMK group had no statistical significance compared with the simvastatin group (P>0.05). The specific value of p-JAK2/JAK2 and p-STAT3/STAT3 protein relative expressions in the simvastatin-treated group (20, 40 mg/L) were significantly lower than that in the control group, respectively (both P<0.05). The specific value of p-JAK2/JAK2 and p-STAT3/STAT3 protein relative expressions in IL-6 group were significantly higher than those in control group (both P<0.05), the specific value of p-JAK2/JAK2 and p-STAT3/STAT3 protein relative expressions in simvastatin (20, 40 mg/L) combined with IL-6 groups were lower than those in IL-6 group (all P<0.05), respectively. Conclusion: Simvastatin can induce the apoptosis of A549 cells through a non-caspase-dependent mitochondrial apoptosis pathway, which may be achieved by inhibiting the JAK2/STAT3 pathway.

[CD7 expression and its prognostic significance in acute myeloid leukemia patients with wild-type or mutant CEBPA].

Objective: To analyze the prognostic value of CD7 expression in newly diagnosed acute myeloid leukemia (AML) patients, and to further explore the correlation between CD7 expression and CEBPA mutation, and to clarify the prognostic value of CD7(+) in AML patients with wild-type (WT) or mutant-type (MT) CEBPA. Methods: The clinical data of 298 newly diagnosed non-M(3) AML patients between January 2010 and December 2016 were analyzed retrospectively. The clinical characteristics and prognosis of CD7(+) and CD7(-) patients were respectively compared in all patients, and in patients with WT and MT CEBPA. The relationship between CD7 expression and CEBPA mutation was determined by chi-square, and the effects of CEBPA mutation on survival and prognosis in CD7(+) group by Kaplan-Meier method. Results: In CD7(+) group, the frequencies of CEBPA mutation were 10.1% (single site) and 33.9% (double site) , significantly higher than those of the CD7(-) group (5.3% and 4.2%) (P=0.000) . Subgroup prognostic analysis showed a lower CR rate (P=0.001) and a higher RR (P=0.023) in CD7(+) group comparing to those of CD7(-) group in AML patients with wild type CEBPA. There were no statistical difference between CD7(+) group and CD7(-) group in overall survival (OS) and disease free survival (P>0.05) , while in the CEBPA mutant group the CD7(+) group has higher OS (P=0.019) and DFS (P=0.010) . Based on the CD7 expression and CEBPA mutation, 298 cases were divided into 3 subgroups, named as CD7(+)-CEBPA MT group, CD7(-) and CD7(+)-CEBPA WT group. The 3-year OS of the 3 groups were 80.2%, 48.0% and 30.6%, respectively (P<0.001) , and the 3-year DFS were 74.1%, 37.4% and 22.2%, respectively (P<0.001) . Conclusion: The CEBPA mutation rate was higher in CD7(+) AML patients then that of CD7(-) patients. CD7 expression has opposite prognostic significance in AML patients carrying the wild-type or mutant-type CEBPA. Based on CD7 expression and CEBPA mutation, a new risk stratification model can be established, which is helpful to guide the clinical individualized treatment for AML patients.

[The expression and significance of chromobox protein homolog 2 in breast cancer].

Objective: To study the relationship between the expression of Chromobox protein homolog (CBX) mRNA and the clinicopathological prognosis of breast cancer, and to investigate the possibility of Chromobox protein homolog 2 as a therapeutic target for breast cancer. Methods: First, we analyzed the mRNA expression of 8 CBX family genes by METABRIC database, and investigated the relationship between the expression of CBX2 mRNA and the clinicopathological parameters of breast cancer. Then we explored its relationship with prognosis. CBX2 siRNA was used to treat breast cancer cell lines with high expression of CBX2(SUM159 and SUM1315). The effects of knockdown of CBX20 on mRNA and protein expression and cell proliferation were observed. Results: According to the analysis of METABRIC database, among the 8 CBX genes, the most obvious increase in mRNA expression was CBX2, and 22.47% (445/1 980) of the patients showed high mRNA expression. The high expression of CBX2 was closely related to tumor histological grade and the molecular type of breast cancer (P<0.001). Compared with the low-expression group of CBX2 mRNA, the proportion of HER2 breast cancer (28.1% vs 7.5%) and Basal-like (44.5% vs 8.5%) in the high-expression group were both higher. Patients with high CBX2 expression had significantly shorter disease-free survival (DFS) and overall survival (OS). The knockdown of CBX2 by siRNA inhibited the proliferation of breast cancer cells. Conclusion: CBX2 is closely related to the prognosis of breast cancer and may be a target for breast cancer treatment.

A study of damage control theory in the treatment of multiple trauma mainly represented by emergency abdominal trauma.

OBJECTIVE: This study aimed to explore the measures of damage control theory (DCT) in the treatment of multiple trauma mainly represented by emergency abdominal trauma. PATIENTS AND METHODS: A total of 76 patients with severe multiple trauma in the Yiwu Central Hospital were selected. Among them, 37 patients with severe multiple trauma were treated with DCT (DCT group), and 39 patients were treated with traditional methods (control group). The prothrombin time (PT), the inflammation index, the duration of mechanical ventilation, the length of stay in the Intensive Care Unit (ICU), and the incidence of sepsis were compared between the two groups. RESULTS: A total of 60 cases (78.95%) were cured and discharged, and 4 cases (10.81%) died in the DCT group, while 12 cases (30.77%) died in the control group. There were 6 cases (16.22%) of sepsis in the DCT group and 15 cases (38.46%) of sepsis in the control group. This indicates that the mortality and the incidence of sepsis in the DCT group were lower than those in the control group (p<0.05 in all comparisons). The PT activated partial thromboplastin time (APTT), the length of stay in the ICU, and mechanical ventilation in the DCT group were notably shorter than those in the control group. The levels of serum tumor necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6), C-reactive protein (CRP), and IL-10 went up remarkably in both groups (p<0.05), but the levels of serum TNF-α, IL-6, and CRP in the DCT group were lower than those in the control group, while the IL-10 level in the former was significantly higher than that in the latter (p<0.05). CONCLUSIONS: It is feasible to apply DCT to rescue patients with multiple trauma, which can effectively reduce the mortality and complications, and shorten the length of stay in the ICU.

[The influence factors of pathologic complete response of the ipsilateral supraclavicular lymph node of breast cancer after neoadjuvant chemotherapy].

Objective: To investigate the factors affecting the pathologic complete response (PCR) of the ipsilateral supraclavicular lymph node (ISLN) of breast cancer after neoadjuvant chemotherapy (NAC). Methods: A total of 178 patients with breast cancer who had primary ipsilateral supraclavicular lymph node metastasis (ISLNM), receiving NAC and subsequent ISLN dissection, were retrospectively reviewed. The single factor and multi factor analysis were carried out by the chi square test and the Logistic regression model. Results: The enrolled patients were all female, 28 to 74 years old. The rate of PCR on the ISLN was 52.2%. Single factor analysis showed that KI67 expression level (χ(2)=7.717,P=0.005), breast PCR (bPCR) (χ(2)=33.564,P<0.001), and axillary PCR (aPCR) (χ(2)=31.750, P<0.001) were associated with the ISLN PCR. Multifactor analysis showed that KI67 expression level (OR=4.096, 95%CI: 1.176-14.263, P=0.027), bPCR (OR=4.452, 95%CI: 1.894-10.461, P<0.001) and aPCR (OR=5.183, 95%CI: 1.974-13.605, P<0.001) were independent predictors of ISLN PCR. The rate of PCR on the ISLN was 90.9% in the patients with KI67>30% and simultaneous breast and axilla PCR. Conclusions: The PCR rate of the ISLN after neoadjuvant chemotherapy is higher than that of the breast and axillary PCR. The expression level of KI67, the bPCR and the aPCR are independent predictors of the PCR on the ISLN.

[Pharmacological effects of site specific conjugated anti-human epidermal growth factor receptor 2-antibody drug conjugate using unnatural amino acid technology].

OBJECTIVE: To investigate inhibitory activities of a homogenous anti-human epidermal growth factor receptor 2 (HER2)-antibody drug conjugate (ADC) on the proliferation of nine tumor cell lines with different levels of HER2 expressions, and its activities on the tumor growth of five xenograft mouse models. METHODS: The HER2 expression levels of BT-474, Calu-3, MCF-7, MDA-MB-231, MDA-MB-468, SK-BR-3, SK-OV-3, HCC1954, NCI-N87 tumor cell lines were measured using QIFI KIT. For the in vitro anti-proliferation assay, serial diluted anti-HER2-ADC, ado-trastuzumab emtansine, AS269, pAF-AS269 and paclitaxel were added to the seeded cells, and after 72 or 96 hours of incubation, the cell proliferation was analyzed. For the in vivo activity, 5-6 weeks old mice were inoculated with four HER2 positive tumor cell lines HCC1954, BT-474, SK-OV-3, NCI-N87 or one HER2 negative tumor cell line MDA-MB-468. Different amounts of anti-HER2-ADC, ado-trastuzumab emtansine, trastuzumab, paclitaxel and phosphate buffered saline control were injected after the tumor volume reached a certain size, then the tumor growth inhibition was analyzed. RESULTS: The expression levels of the six high HER2-expression cell lines SK-OV-3, NCI-N87, SK-BR-3, Calu-3, HCC1954, BT-474 were between 430 000 to 800 000 receptors per cell, which were 50 times higher than those of the other three low HER2 expression tumor cell lines MDA-MB-231, MCF-7, MDA-MB-468. Anti-HER2-ADC had inhibition effects on cell lines with high level of HER2 expression in the in vitro anti-proliferation assay. The half maximal inhibitory concentrations of anti-HER2-ADC on SK-OV-3, NCI-N87, SK-BR-3, Calu-3, HCC1954, BT-474 tumor cell lines were 46 pmol/L, 17 pmol/L, 17 pmol/L, 161 pmol/L, 125 pmol/L, 50 pmol/L, respectively. Anti-HER2-ADC had a dose dependent antitumor activity in vivo in all the HER2 positive xenograft mouse models. In NCI-N87 xenograft tumor model, the same dose of anti-HER2-ADC showed better anti-tumor activity compared with trastuzumab and ado-trastuzumab emtansine, and its relative tumor proliferation rates were about 1/30 to 1/20 of the two. In HCC1954 xenograft tumor model, the complete regression of the tumor was observed. As expected, anti-HER2-ADC had no tumor inhibitory effects on MDA-MB-468 xenograft models with low HER2 expression. The antitumor activities of anti-HER2-ADC in HER2 positive xenograft tumor models were the same as or better than the activities of ado-trastuzumab emtansine. CONCLUSION: The homogenous site-specific anti-HER2-ADC obtained using unnatural amino acid technology can inhibit the growth of high HER2-expression tumor cells with high potency both in vivo and in vitro.

[Analysis of factors influencing the axillary lymph node metastasis and breast cancer-specific survival in patients with T1 breast cancer].

Objective: To investigate the predictors of axillary lymph node metastasis and the breast cancer-specific survival (BCSS) in patients with T1 breast cancer. Methods: A retrospective analysis of clinical and pathological data of 840 T1 invasive breast cancer cases between January 2009 and January 2014 in Henan Cancer Hospital was conducted.Chi square test and Logistic regression analysis were carried out to identify relevant factors of lymph node metastasis. Analysis of prognostic factors were analyzed by Log-rank test and Cox regression. Results: Among the 840 T1 breast cancer cases, positive axillary lymph nodes were found in 150 (17.9%) cases. Univariate analysis showed that tumor size, histological grade, tumor location, and HER2 status were associated with axillary lymph node status (P<0.05). Multivariate analysis showed that tumor size, histological grade, tumor location, and HER2 status were independent predictive factors of axillary lymph node metastasis (P<0.05). Log-rank test showed that tumor size, histological grade, HER2 status, partial response (PR) status and number of positive lymph nodes were important factors influencing BCSS of the patients with positive axillary lymph nodes (P<0.05). Cox analysis showed that the size of the primary tumors and the number of positive lymph nodes were independent factors affecting the BCSS of the patients(P<0.05). Conclusions: Tumor size, histological grade, tumor location and HER2 status correlated with axillary lymph nodes status of T1 breast cancer. For T1 breast cancer patients with positive axillary lymph node, more positive lymph nodes involved and smaller primary tumor correlated with worse prognosis.

[Screening for hotspot mutations associated with genetic hearing impairment in pregnant women and subsequent prenatal diagnosis in high risk pregnancies].

Objective: To screen for hotspot gene mutations associated with genetic deafness in Chinese pregnant women, and to perform risk assessment and prenatal diagnosis in high-risk families. Methods: Between November 2012 and October 2017, 26 117 pregnant women were screened by molecular hybridization microarray for 9 hot-spot mutations in 4 hereditary deafness related genes (GJB2 c. 35 del G, c. 176_191 del 16 bp, c. 235 del G, c. 299_300 del AT, GJB3 c. 538 C>T, SLC26A4 c. 2168 A>G, IVS 7-2 A>G, mitochondrial DNA 12S rRNA m. 1494 C>T, m. 1555 A>G). Genotype analysis was carried out in husbands of women carrying mutations, and prenatal diagnosis was carried out in the fetuses with high risk of deafness. Results: Among all women tested, 1 208(4.63%) were carriers of genetic deafness mutations, 7 with hearing impairment were affected by homozygous or compound heterozygous mutations, 51 were mitochondrial gene mutation carriers, 103 were carriers of GJB3 c. 538 C>T heterozygous mutation, 1 026 were carriers of GJB2 or SLC26A4 heterozygous mutations, and 21 carried heterozygous mutations in two genes simultaneously. In 394 families, the husbands accepted gene sequence testing, and 27 in which were determined as carriers of mutations in identical genes as their wives. Among which, 18 families received prenatal diagnosis, and 5 fetuses were diagnosed as hereditary deafness. In 9 families who did not receive prenatal diagnosis, 1 neonate was diagnosed as compound heterozygote after delivery. Conclusion: In order to prevent birth defects with congenital hearing problems, it is effective to provide screening for hotspot mutations in pregnant women and to perform prenatal diagnosis on high risk pregnancies.

[The influence of lumpectomy on the axillary lymph node status of breast cancer patients].

Objective: To investigate the influence of lumpectomy on axillary lymph node status of breast cancer patients. Methods: The clinical data of 738 invasive breast cancer patients with non-palpable axillary lymph node and sentinel lymph node (SLN) biopsy from November 2011 to August 2013 in Henan Provincial Cancer Hospital were collected and retrospectively analyzed. Among them, 136 patients underwent preoperative lumpectomy (lumpectomy group) and 602 patients underwent puncture biopsy only (biopsy group). The difference of axillary lymph node status and positive ratio of SLN detected by color Doppler ultrasound were compared between these two groups. Results: Among the 738 breast cancer patients, the axillary lymph nodes of 444 (60.2%) cases could be detected by ultrasound. Among them, 92 cases belonged to lumpectomy group, significantly less than 352 cases of biopsy group (P=0.048). Among the patients with ultrasound-visible lymph nodes, the proportion of the biggest diameter of axillary lymph node >1 cm of lumpectomy group or biopsy group was 58.7% (54/92) or 52.8% (186/352), respectively, without significant difference (P=0.316). The proportion of patients with the ratio of long diameter to short diameter <2 of lumpectomy group or biopsy group was 37.0% (34/92) or 38.6% (136/352), respectively, with marginal difference (P=0.768). The positive rate of SLN of lumpectomy group or biopsy group was 23.5% (32/136) or 26.9% (162/602), respectively, without significant difference (P=0.419). The incidence rate of the ultrasound visible axillary lymph nodes of patients whose postoperative time ≤ 7 days or > 7days was 71.1% (64/90) or 60.9% (8/46), respectively, without significant difference (P=0.227). However, the positive rate of SLN of these two groups was 28.9% (26/90) and 13.0% (6/46), respectively, with significant difference (P=0.039). The number of ultrasound visible axillary lymph nodes, the biggest diameter of axillary lymph nodes and the ratio of the long diameter to short diameter <2 were substantially correlated with the positive rate of SLN (P<0.05). Conclusions: The incidence rate of ultrasound visible axillary lymph node in the patients with lumpectomy is higher than that of patients with puncture biopsy only. The positive rate of SLN of the patients with a long postoperative time is lower than that of patients with a short postoperative time, even though the axillary lymph nodes are ultrasound visible.

Systemic mastocytosis identified in two women developing fragility fractures during lactation.

Two women presenting with fragility fractures during lactation had bone mineral density (BMD) reduced more greatly than usually associated with lactation. The first woman was 29 years old with a BMD T-score of - 3.2 SD at the spine and- 2.0 SD at the femoral neck. The second woman was 35 years old with a BMD T-score of - 4.5 SD at the spine and - 2.8 SD at the femoral neck. Both women had increased cortical porosity and reduced trabecular density. Investigation identified an elevated serum tryptase, and marrow biopsy confirmed the diagnosis of mastocytosis. Lactation causes bone loss, but the occurrence of fractures in the setting of severe deficits in BMD and microstructural deterioration signals the need to consider additional causes of bone loss.

[Analysis of predictive effect of Androgen receptor on the response to neoadjuvant chemotherapy in breast cancer patients].

Objective: To explore the expression of androgen receptor (AR) in the tissues as well as its association with the clinicopathological factors of primary breast cancer patients treated with neoadjuvant chemotherapy (NAC), and analyze the effect of AR in the prediction of pathologic complete response (PCR) rate. Method: A total of 668 breast cancer patients treated with NAC in Henan Cancer Hospital between March 2014 and June 2017 were retrospectively reviewed. The relationship of AR expression and clinicopathological characteristics was calculated using chi square test. Multivariate analysis using binary Logistic regression was used to analyze correlations of different factors with PCR. Result: All patients were female, with the age of 20-76 years old. AR was detected in 74.6% of tumors, and significantly correlated with hormone receptor (HR), human epidermalgrowth factor receptor-2 (HER-2), Ki-67, CK5/6, epidermal growth factor receptor (EGFR) and molecular subtypes (all P<0.05). Multivariate analysis showed that AR, HR and HER-2 were independent predictors for PCR (all P<0.05). Conclusions: The expressions of AR were more frequently in HR positive breast cancer tissues (86.7%), and lowest in triple-negative breast cancer (TNBC) group (23.2%). AR was independent predictor for PCR.

MiR-101 promotes nasopharyngeal carcinoma cell apoptosis through inhibiting Ras/Raf/MEK/ERK signaling pathway.

OBJECTIVE: Extra-cellular signal regulated kinase (ERK)/mitogen activated protein kinase (MAPK) signaling pathway is widely involved in cell proliferation and apoptosis. MAPK kinase 1 (MEK1) is the upstream protein kinase of ERK that can activate ERK/MAPK signaling pathway. microRNA-101 (MiR-101) down-regulation is found to be associated with nasopharyngeal carcinoma (NPC) pathogenesis. Bioinformatics analysis shows the complementary targeted relationship between miR-101 and the 3'-UTR of MEK1 mRNA. This study explores the role of miR-101 in regulating MEK1 expression, ERK/MAPK signaling pathway activation, and NPC pathogenesis. MATERIALS AND METHODS: Dual luciferase assay confirmed the targeted relationship between miR-101 and MEK1. MiR-101 and MEK1 expressions were compared in inflammatory nasopharynx tissue and NPC tissue. MiR-101, MEK1, phosphorylated ERK 1/2 (p-ERK1/2), survivin expressions in NP69, CNE-1, HONE1, and C666-2 cell lines were detected. NPC cell line C666-1 was cultured in vitro and divided into four groups, including miR-NC, miR-101, si-NC and si-MEK1. Cell apoptosis was determined by flow cytometry. Cell proliferation was evaluated by EdU staining. RESULTS: MiR-101 targeted inhibited MEK1 expression. MiR-101 was significantly down-regulated, while MEK1 was significantly elevated in NPC tissue compared with inflammatory nasopharynx tissue. MiR-101 was markedly declined, whereas MEK1, p-ERK1/2, and survivin were apparently increased in CNE-1, HONE1, and C666-1 cells compared with NP69 cells. MiR-101 mimic and/or si-MEK1 transfection significantly reduced MEK1, p-ERK1/2, and survivin levels, attenuated cell proliferation, and enhanced cell apoptosis. CONCLUSIONS: Down-regulation of miR-101 was related to NPC pathogenesis. MiR-101 elevation suppressed NPC cell proliferation and promoted apoptosis through targeted inhibiting MEK1 expression to alleviate ERK/MAPK signaling pathway and survivin expression.