Triplane technique for breast reconstruction after breast cancer surgery: A case series report.

RATIONALE: Implant-based breast reconstruction is an important method for post-mastectomy breast reconstruction. Currently, the most commonly used technique is the biplane technique. However, the high rate of postoperative complications, the inability of pockets to accommodate larger implants, and the expensive costs of biological mesh make the development of new surgical methods urgent. The triplane technique for breast reconstruction is an ideal candidate method. PATIENT CONCERNS: The main local symptoms were breast lump, abnormal breast skin, nipple discharge, and abnormal nipple or areola in 24 patients. DIAGNOSES: The study included 24 female patients who underwent breast reconstruction using the triplane technique after radical breast cancer surgery. INTERVENTIONS: The surgical procedure involved measuring the dimensions of the breast, designing the incision, and creating a pocket for the implant using the triplane technique, which includes the pectoralis major muscle, the pectoralis major fascia continuing to the rectus abdominis fascia, and the latissimus dorsa muscle fascia continuing to the rectus abdominis fascia. Postoperative follow-up included regular assessments of pain and evaluation of breast appearance. OUTCOMES: No cases of postoperative infection were observed in all patients. During the 1-year follow-up period after surgery, 5 patients (20.8%) who needed radiotherapy after mastectomy for cancer showed slight darkening of skin flap pigment after using the triplane technique implant. No cases of exposure or infection of the expanders were reported, and 1 patient underwent expander replacement with a permanent prosthesis. All patients expressed satisfaction with the reconstructed breast shape. The 10 patients (41.7%) experiencing postoperative swelling and pain. However, the pain gradually subsided during the postoperative recovery period. No cases of local recurrence or distant metastasis of breast cancer were observed during the 1-year-follow-up period. LESSONS: The triplane technique for breast reconstruction after breast cancer surgery provides good implant coverage, reduces the risk of complications, and is cost-effective.

Systematic review and meta-analysis of imaging differential diagnosis of benign and malignant ovarian tumors.

Background: With the increasing incidence of gynecological ovarian tumors, the differential diagnosis of benign and malignant ovarian tumors is of great significance for subsequent treatment. Currently, ovarian examinations commonly use computed tomography (CT) or magnetic resonance imaging (MRI). This study sought to compare the value of CT and MRI in differentiating between benign and malignant ovarian tumors. Methods: The PubMed, Cochrane Central Register of Controlled Trials, Embase, Web of Science, China National Knowledge Infrastructure, Wanfang, and Weipu databases were searched for published articles using the following terms "CT" or "Computed Tomography" or "MRI" or "Magnetic Resonance imaging" and "ovarian cancer" or "ovarian tumor" or "ovarian neoplasm" or "adnexal mass" or "adnexal lesion". The articles were screened and the data were extracted based on the inclusion and exclusion criteria. The Quality Assessment of Diagnostic Accuracy Studies-2 recommended by the Cochrane Collaboration was used to assess the methodological quality of the included studies, and the network meta-analysis was performed by Stata 15.0. Results: The results showed that the overall sensitivity and specificity of CT were 0.79 [95% confidence intervals (CI): 0.70-0.87] and 0.87 (95% CI: 0.80-0.92), respectively. The overall sensitivity and specificity of MRI were 0.94 (95% CI: 0.91-0.95) and 0.91 (95% CI: 0.90-0.93), respectively. The area under the curve of the CT and MRI summary receiver operating characteristics were 0.9016 and 0.9764, respectively. The positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio of CT were 5.26 (95% CI: 2.78-9.93), 0.26 (95% CI: 0.13-0.50), and 22.19 (95% CI: 7.54-65.30), respectively. The positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio of MRI were 8.69 (95% CI: 5.06-14.92), 0.07 (95% CI: 0.04-0.13), and 146.19 (95% CI: 68.88-310.24), respectively. Conclusions: Compared to CT, MRI has a stronger ability to differentiate between benign and malignant ovarian tumors. It's a promising non-radiological imaging technique and a more favorable choice for patients with ovarian tumors. However, in the future, large-sample, multi-center prospective studies need to be conducted to compare the performance of MRI and CT in distinguishing between benign and malignant ovarian tumors.

Significant growth inhibition by a bispecific affibody targeting oncoprotein E7 in both HPV16 and 18 positive cervical cancer in vitro and in vivo.

The infection with HPV 16 and 18 high-risk types account for more than 80 % of cervical cancer incidence, but there is still no targeted agent against HPV for cervical cancer therapy. Our previous study constructed a bispecific affibody Z16-18 targeting HPV16 and 18 early antigen 7 (E7, responsible for the infected cell malignant transformation). In the present study, we prepared Z16-18 in prokaryotic expression system and confirmed its significant growth inhibition both on SiHa (HPV16 positive) and HeLa (HPV18 positive) cervical cancer cells by arresting cell cycle at G0/G1 phase. The IC50 of Z16-18 on SiHa and HeLa were close in value. Z16-18 could specifically target E7 in both SiHa and HeLa, and exhibited prominent targeted enrichment on tumor tissues derived from SiHa or HeLa, resulting in the inhibition of tumourigenesis and tumour growth in vivo. Furthermore, Z16-18 could inhibit the interaction between E7 and pRb to block the E7-pRb carcinogenic pathway, resulting in the decreased release of E2F and the cell growth inhibition characterized by the decrease of CDK6 and Cyclin D1. This study provides a new strategy for targeted therapy based on affibody, and Z16-18 has great potential for utilisation and development as an agent targeting HPV16 and HPV18 related cervical cancer.

Targeted biological effect of an affitoxin composed of an HPV16E7 affibody fused with granzyme B (ZHPV16E7-GrB) against cervical cancer in vitro and in vivo.

BACKGROUND: High risk type 16 of human papillomavirus (HPV16) is associated with 50% of cervical cancer, for which reliable targeted therapies are lacking. HPV early protein 7 (E7) is an oncoprotein responsible for cell malignant transformation. In our previous work, a highly specific affibody targeting HPV16E7 (ZHPV16E7) was developed. OBJECTIVE: In order to improve the targeted therapeutic effect, the present study prepared an affitoxin consisting of ZHPV16E7 fused with granzyme B (GrB), namely, ZHPV16E7-GrB, and evaluated its targeting action in vitro and in vivo. METHODS: The ZHPV16E7-GrB fusion protein was produced in a prokaryotic expression system. The targeted binding properties of the ZHPV16E7-GrB to the HPV16E7 were confirmed by immunofluorescence assay (IFA) in cervical cancer cell lines, by immunohistochemical assay (IHA) in cervical cancer tissue from clinical specimens and by near-infrared imaging in tumour-bearing mice. The anti-tumour effect on both cervical cancer cells in vitro and tumour-bearing mice in vivo were further evaluated. RESULTS: A 34-kDa ZHPV16E7-GrB fusion protein was produced in E. coli and displayed corresponding immunoreactivity. IFA revealed that ZHPV16E7-GrB bound specifically to HPV16-positive TC-1 and SiHa cells. IHA showed that ZHPV16E7-GrB also bound specifically to HPV16-positive clinical tissue specimens. In addition, the near-infrared imaging results showed that ZHPV16E7-GrB was enriched in tumour tissues. Moreover, both the ZHPV16E7-GrB affitoxin and ZHPV16E7 affibody (without GrB) significantly reduced the proliferation of cervical cancer cells in vitro and tumour-bearing mice in vivo, and the antiproliferative effect of ZHPV16E7-GrB was higher than that of the ZHPV16E7 affibody. CONCLUSIONS: The affitoxin by coupling the affibody with GrB is a promising targeted therapeutic agent with the dual advantages of the targeted affibody and the GrB cytotoxin.

The Roles of Alternative Splicing in Tumor-immune Cell Interactions.

Alternative splicing (AS) plays a significant role in the hallmarks of cancer and can provide neoantigens for immunotherapy. Here, we summarize recent advances in immune system associated tumor specific-antigens (TSAs) produced by AS. We further discuss the regulating mechanisms involved in AS-mediated innate and adaptive immune responses and the anti-tumoral and protumoral roles in different types of cancer. For example, ULBP1_RI, MLL5Δ21spe, NKp44-1Δ5, MHC-IΔ7, CD200SΔ1, 2, PVR α/ß/γ/δ and IL-33 variants 1/2/3 act as regulators in solid tumors and IPAK4-L and, FOXP1ΔN100 exhibit functions in hematological cancers.

Identification of Novel Compound Heterozygous Mutations in the GAN Gene of a Chinese Patient Diagnosed With Giant Axonal Neuropathy.

Giant axonal neuropathy (GAN) is a very rare autosomal recessive disorder characterized by abnormally large and dysfunctional neuronal axons. Mutations in the GAN gene have been identified as the cause of this disorder. In this report, we performed a detailed phenotypic assessment of a Chinese patient with GAN. An array-based exon capture test and targeted next-generation sequencing were used to detect the suspected mutation sites. Compound heterozygous mutations of p.S79L (c.236C > T) in the BTB domain and p.T489S (c.1466C > G) in the kelch domain were identified in the proband's genome. S79L was a known mutation, and T489S was reported for the first time. The p.S79L and p.T489S were confirmed in the proband's mother and father, respectively. Both mutations were located in highly conserved regions and affected the predicted protein crystal structures. The proband's sural biopsy revealed the classical GAN phenotype of swollen axons filled with closely packed neurofilaments. The combined application of the next-generation sequencing platform and bioinformatics analyses was an effective method for diagnosing GAN. The novel compound mutations of S79L and T489S in the GAN gene were likely the cause of the patient's GAN symptoms. Our findings enrich the spectrum of mutations associated with this rare type of axonopathy.

The Effect of Mitomycin C on Reducing Endometrial Fibrosis for Intrauterine Adhesion.

BACKGROUND Intrauterine adhesion (IUA) is a common reproductive system disease in women, characterized by endometrial stromal cell proliferation, increasing fibroblasts and increasing extracellular matrix secretion. The purpose of this study was to investigate the effect of mitomycin C on reducing endometrial fibrosis for IUA. MATERIAL AND METHODS Firstly, a rat IUA model was constructed by intrauterine mechanical injury. The endometrial stromal cells and fibroblasts were isolated and treated with mitomycin C. After that, Cell Counting Kit-8 (CCK-8) assay was used to investigate the endometrial stromal cell viability. Furthermore, cell cycle and apoptosis assays of endometrial stromal cells and fibroblasts were performed, respectively. Finally, the cell viability of human endometrial cells or human uterus adhesion fibroblasts treated with mitomycin C was determined using CCK-8 assay with or without estradiol. RESULTS Endometrial stromal cells were isolated from a rat IUA model. Cell cycle assay results showed that mitomycin C inhibited cell viability and promoted G1 cell cycle arrest and apoptosis in rat IUA endometrial stromal cells. Fibroblasts were also isolated from the rat IUA model. We found that mitomycin C inhibited the synthesis and secretion of collagen type I by western blotting analysis. Furthermore, mitomycin C promoted G1 cell cycle arrest and apoptosis in IUA rat uterine fibroblasts. We found that estradiol decreased the inhibitory effects of cell viability of human endometrial cells and human uterus adhesion fibroblasts by mitomycin C. CONCLUSIONS Our findings revealed that mitomycin C could reduce endometrial fibrosis for intrauterine adhesion.

Effective Neutralizing Antibody Produced in Mice Directly Immunized with Integrated Pichia pastoris Expressing HPV16L1 Protein.

The human papillomavirus (HPV) vaccine has not been widely used in developing countries because of its high cost and multiple subtype restrictions. The present study aimed to develop an economical, convenient, and effective vaccine to produce neutralizing antibodies. Using late protein 1 (L1) from the HPV16 subtype as the target antigen (HPV16L1) and Pichia pastoris as the antigen release system, integrated P. pastoris expressing HPV16L1 (named yeast-HPV16L1) was prepared and vaccinated directly into mice by subcutaneous multipoint injection. After immunization was performed thrice, high titers (greater than 1:40,960) of specific anti-HPV16L1 antibodies were obtained in immune serum and were observed to continuously rise over time. The indirect hemagglutination test and indirect hemagglutination inhibition test were used to detect neutralizing antibody activity in vitro, and the results demonstrated the hemagglutination ability of the immune serum and the reduction in or loss of the hemagglutination ability if preneutralized antigen was added to the immune serum. The protection conferred by immune serum to tumor-bearing mice at the early stages was confirmed, but the neutralizing activity disappeared when the tumor reached a size of 1 mm3. The neutralization activity of the immune serum was confirmed both in vitro and in vivo.

Evaluation of ovarian function using three dimensional ultrasound in perimenopausal women.

To evaluate the feasibility and clinical value of three-dimensional ultrasound in evaluating ovarian function in perimenopausal women. In this prospective cohort study, 102 patients with clinically suspected perimenopause and 90 patients with menopause were enrolled. These patients were classified into three groups according to the level of follicle stimulating hormone (FSH) and estradiol (E2): menopause group, perimenopause group, and normal group. Perimenopause group: There were significant differences in volume, vascularization index (VI), flow index (FI), and vascularization-flow index (VFI) in the ovaries after treatment. Cycle 1 > cycle 0 (p < .05) and cycle 3 cycle 0 (p < .05), and in FSH: cycle 3 < cycle 0 (p < .05). Three-dimensional ultrasound in ovarian quantitative measurement can objectively reflect the change in the ovarian function, predicting the effect of drug treatment, and provided an objective information for early intervention to menopausal.

Development of biodegradable polyesters based on a hydroxylated coumarin initiator towards fluorescent visible paclitaxel-loaded microspheres.

In this work, we developed a facile end-functionalization method using hydroxylated coumarin to initiate the ring-opening polymerization of cyclic esters to synthesize a series of fluorescent biodegradable aliphatic polyesters with tailorable properties. The resulting fluorescent functionalized poly(l-lactide) (PLLA-COU), poly(ε-caprolactone) (PCL-COU) poly(δ-valerolactone) (PVL-COU) and poly(trimethylene carbonate) (PTMC-COU) were investigated to evaluate the dependence of fluorescence on the chemical structure and molecular weight of the materials. The differences in the electron withdrawing ability and the density of ester groups are responsible for the changes in the fluorescence quantum yield. Then, two representative biodegradable materials, namely, PLLA-COU and PCL-COU, were used to prepare fluorescent paclitaxel-loaded microspheres. During in vitro drug release, the release rate of the PCL-COU microspheres is dramatically faster than that of the PLLA-COU microspheres due to the difference in the material nature and their surface morphologies, possibly achieving a tunable degradation and release rate for the drug carriers. Fluorescent functionalized polyester microspheres can retain their fluorescence properties and emit bright blue light for fluorescence tracing during the degradation process. Biological evaluations showed that both fluorescent polyesters are devoid of any significant toxicity and have good biocompatibility. The results demonstrated that the obtained fluorescent polyesters are promising for use in traceable and controlled drug delivery with tunable drug release.

The application of methylene blue coloration technique in axillary lymph node dissection of breast cancer.

BACKGROUND: To analyze the associated factors that affect the coloration of methylene blue (MB) in axillary lymph node dissection (ALND) of patients with breast cancer and to explore the tracer effect of MB in high axillary lymph node metastasis, to guide surgical treatment. METHODS: We recruited 170 patients who underwent ALND, all of them were injected MB before operation. We analyzed the relationships between the clinical factors of age, body mass index (BMI), molecular typing, TNM staging, neoadjuvant chemotherapy, injection time and position and the MB coloration by univariate and multivariate analyses. A total of 84 cases were selected for observation of the application value of MB during intervention involving the lymph nodes upper axillary vein. RESULTS: Of 170 cases, 138 cases (81.17%) were colored. Univariate analysis showed that significant differences were observed between differing BMI's (χ2=24.074, P<0.0001) and injection times (χ2=41.207, P<0.0001). Multivariate analysis showed that injection time (P=0.016) was the clinical factor associated with MB coloration. More than 60 minutes before surgery and MB injection, the possibility of methylene blue colored was 0.088 times higher than 10 minutes (P=0.010, 95% CI, 0.014, 0.554). MB was used to track the lymph node upper axillary vein with a sensitivity of 12.5%, a specificity of 100%, a false negative rate of 87.50%, and a false positive rate of 0; the kappa coefficient was 0.051 (χ2=0.679, P=0.404). CONCLUSIONS: Among the clinical factors, MB coloration was worse more than 60 minutes before ALND. Using the technology of MB color, lymph nodes can be clearly identified. It has great guiding value for the doctors who learn the operation initially. However, it is still prudent to use MB for high lymph node dissection.

Induction of Humoral and Cellular Immune Responses in Mice by Multiepitope Vaccines Composing of Both T and B Lymphocyte Epitopes of MAGE-A3 which are Recombined into HBcAg.

BACKGROUND: Melanoma-associated antigen-A3 (MAGE-A3) is a tumor specific antigen and a potential candidate for cancer immunotherapy. We had screened three immunodominant multiepitopes of MAGE-A3, and identified these multiepitope peptides had significantly higher reactivity to serum samples from gastric cancer patients. However, the immune responses of three multiepitope peptides carried by HBcAg in mice have not been investigated. OBJECTIVES: The main objective of this study was to analyze the humoral and cellular immune responses in mice induced by these three multiepitope vaccines of MAGE-A3. METHODS: Three multiepitopes of MAGE-A3 (MAGE-A3(EPI-1, or -2, or -3)) were respectively inserted at HBcAg major immunodominant region (HBcAg(MIR)) of the pET21a(+)/HBcAg(MIR) recombinant plasmid. These recombinant chimeras were identified by PCR, and transfected respectively into E. Coli Ressotta strain. The expression products of rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) were purified respectively by Ni2+ chelated affinity column, and then confirmed by SDS-PAGE and Western-blot analysis.Purified three rHBcAg(MIR)/MAGE-A3 multiepitopes were administrated respectively into BALB/c (H-2Kd) mice by intradermal injection. The production of rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) specific IgG in serum from immunized mice were measured by ELISA. Spleen cells from all immunized mice were harvested after one week of last immunization for lymphocyte proliferation assay and cytotoxic T-lymphocyte assay. RESULTS: PCR and Sequencing analysis showed the presence of the required gene fragment in pET21a(+)/ HBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) recombinant plasmid. Purified rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) could be probed specifically by McAb of 6×his-tag. ELISA analysis indicated that serum from immunized mice with rHBcAg(MIR)/MAGE-A3(EPI-1, -2, or -3) proteins could be discerned specifically by complete MAGE-A3 protein, and high level of antibodies in immune serum were obtained, and all antibody titers could reach above 1:1600. The splenocytes from groups of rHBcAg(MIR)/MAGE-A3(EPI-1,-2, or -3), stimulated respectively with corresponding peptides showed the higher proliferative responses comparing with control groups of HBcAg(MIR) or PBS (p<0.05, respectively). Splenocytes from mice immunized with rHBcAg(MIR)/MAGE-A3 (EPI-1, or -2, or -3) could killed target cells effectively, and there were significant difference of CTL activities compared with control groups of HBcAg(MIR), or PBS (p<0.05, respectively) at any ratio of effector : target. CONCLUSION: Our results indicated MIR in HBcAg presenting platform could present MAGE-A3 multiepitopes efficiently and induced significant humoral or cellular immunity. The immune strategy based on multiepitopeimmunization could have potential for preventing or controlling MAGE-A3 associated malignant disease.

[Expression of VEGF-C and VEGF-D in gastric carcinoma and its relationship with lymph node metastases].

OBJECTIVE: To explore the expression of vascular endothelial growth factor (VEGF) C and D in gastric cancer and its relationship with tumor angiogenesis and lymph node metastasis. METHODS: Immunohistochemistry(SABC) and real-time PCR were used to detect the expression of VEGF-C, VEGF-D protein and mRNA in 32 gastric cancer tissues and 32 normal gastric tissues. RESULTS: The positive expression rate of VEGF-C and VEGF-D in gastric cancer tissue was significantly higher than that of normal gastric tissues (P<0.01), the expression of VEGF-C and VEGF-D in the gastric cancer group and the lymph node metastasis group were significantly different (P<0.05). The expression of VEGF-C and VEGF-D in gastric carcinoma was positively correlated with lymph node metastasis (P<0.01), the expression of VEGF-C and VEGF-D in well-differentiated carcinoma, moderately differentiated carcinoma and poorly differentiated carcinoma was statistically different (P<0.05). VEGF-C and VEGF-D expressions in gastric cancer cells were not related to the patient's age, sex, and lymph node distant metastasis (P>0.05). CONCLUSION: The non-intake high expression of VEGF-C and VEGF-D in gastric cancer cells is closely related to lymph node metastasis. They serve as the important reference indicator to assess the prognosis in gastric cancer patients.