Partial two-stage exchange: an alternative method for infected total hip arthroplasty.

BACKGROUND: Total two-stage exchange is commonly used in clinical practice as a treatment for infected total hip arthroplasty (THA); however, this approach involves considerable limitations, including significant bone loss and severe trauma. This retrospective cohort study was conducted to evaluate clinical outcomes following the use of partial two-stage exchange (PTE) for infected THA. METHODS: We performed a retrospective analysis of 28 patients with infected THA who were treated by PTE between September 2000 and June 2019. Eligibility for PTE was limited to patients with a well-fixed femoral stem prosthesis. In the first stage of the operation, the femoral stem prosthesis was preserved; subsequently, the acetabular prosthesis, liner, and head were replaced with an antibiotic-loaded spacer. The new prosthesis was then implanted into patients and monitored for at least 3 months to ensure freedom from infection. RESULTS: Patients were followed for an average of 4 years (range, 2-11 years), with an overall success rate of 85.7% (24/28). The mean Harris hip score at the final follow-up was 76.2 ± 11.7 points. CONCLUSIONS: The findings of this study suggest that PTE could be an acceptable option for a subset of patients with infected THA, offering a satisfactory infection control rate and clinical outcomes comparable to those of total two-stage exchange, but with less harm.

Synthesized natural peptides from amphibian skin secretions increase the efficacy of a therapeutic vaccine by recruiting more T cells to the tumour site.

BACKGROUND: Therapeutic vaccines against cervical cancer remain ineffective. Previously, we demonstrated that blocking the signalling of a cytokine, interleukin 10, at the time of immunisation elicited significantly higher numbers of antigen specific T cells and inhibited tumour growth in mice. RESULTS: In the current paper, we demonstrate, in a HPV16 E6/E7 transformed TC-1 tumour mouse model, that despite increased antigen specific T cell numbers, blocking IL-10 signalling at the time of immunisation does not increase the survival time of the TC-1 tumour bearing mice compared to mice receiving the same immunisation with no IL-10 signalling blockade. Moreover, the function of tumour infiltrating T cells isolated 3 weeks post TC-1 transplantation is more suppressed than those isolated 2 weeks after tumour inoculation. We demonstrate that synthesized caerin peptides, derived from amphibian skin secretions, 1) were able to inhibit TC-1 tumour growth both in vitro and in vivo; 2) are environmentally stable; and 3) promote the secretion of pro-inflammatory interlukine-6 by TC-1 cells. Notably caerin peptides were able to increase the survival time of TC-1 tumour bearing mice after therapeutic vaccination with a HPV16E7 peptide-based vaccine containing IL-10 inhibitor, via recruiting increased levels of T cells to the tumour site. CONCLUSION: Caerin peptides increase the efficacy of a therapeutic vaccine by recruiting more T cells to the tumour site.

Clinical efficacy and safety of cervical intralymphatic immunotherapy for house dust mite allergic rhinitis: A pilot study.

BACKGROUND: Previous studies have demonstrated that intralymphatic immunotherapy (ILIT), a less time-consuming alternative to conventional subcutaneous immunotherapy (SCIT), is safe and effective. However, because of the private location of inguinal lymph nodes, inguinal ILIT is relatively inconvenient. We proposed a novel form of ILIT that involves 3 injections of allergen into cervical lymph nodes. The aim of this study is to determine the clinical efficacy and safety of cervical ILIT on house dust mite induced allergic rhinitis (AR) in adults. METHODS: In this study, we performed a prospective cohort study to determine the clinical efficacy and safety of cervical ILIT on house dust mite induced AR in adults, by comparing the symptom scores, quality-of-life scores (QOLS) and drug scores (use of rescue medication) before and after treatment. Meanwhile, side events were also recorded. RESULTS: Cervical ILIT elicited no moderate-severe adverse events. Patients receiving cervical ILIT experienced a significant improvement in nasal symptoms, eye symptoms and quality of life, as compared to baseline (P all <0.001). A reduction in the use of rescue medication was also demonstrated (P < 0.001). CONCLUSIONS: In this first-in-human clinical study, cervical ILIT was demonstrated safe and induced allergen tolerance after 3 injections.

TCR repertoire profiling of tumors, adjacent normal tissues, and peripheral blood predicts survival in nasopharyngeal carcinoma.

The T-cell immune responses in nasopharyngeal carcinoma patients have been extensively investigated recently for designing adoptive immunotherapy or immune checkpoint blockade therapy. However, the distribution characteristics of T cells associated with NPC pathogenesis are largely unknown. We performed deep sequencing for TCR repertoire profiling on matched tumor/adjacent normal tissue from 15 NPC patients and peripheral blood from 39 NPC patients, 39 patients with other nasopharyngeal diseases, and 33 healthy controls. We found that a lower diversity of TCR repertoire in tumors than paired tissues or a low similarity between the paired tissues was associated with a poor prognosis in NPC. A more diverse TCR repertoire was identified in the peripheral blood of NPC patients relative to the controls; this was related to a significant decrease in the proportion of high-frequency TCR clones in NPC. Higher diversity in peripheral blood of NPC patients was associated with a worse prognosis. Due to the peculiarity of the Vß gene usage patterns in the peripheral blood of NPC patients, 15 Vß genes were selected to distinguish NPC patients from controls by the least absolute shrinkage and selection operator analysis. We identified 11 clonotypes shared by tumors and peripheral blood samples from different NPC patients, defined as "NPC-associated" that might have value in adoptive immunotherapy. In conclusion, we here report the systematic and overall characteristics of the TCR repertoire in tumors, adjacent normal tissues, and peripheral blood of NPC patients. The data obtained may be relevant to future clinical studies in the setting of immunotherapy for NPC patients.

Blocking IL-10 signalling at the time of immunization renders the tumour more accessible to T cell infiltration in mice.

We recently reported that blockade of IL-10 signalling at the time of a human papillomavirus (HPV) long E7 peptide/LPS immunization leads to the regression of established HPV-16 immortalized tumours in mice similar to that induced by long E7 peptide/incomplete Freund's adjuvant (IFA)-based vaccination. In this paper, we demonstrated that blockade of IL-10 signalling at the time of long E7 peptide/LPS could elicit stronger T cells responses and render the tumour more accessible for immune cell infiltration than vaccination with long E7 peptide/IFA. Furthermore, priming with long E7 peptide/LPS and IL10 signalling blockade then boosting with long E7 peptide/IFA elicits stronger CD8+ T cell responses than long E7 peptide/IFA immunization. The results suggest that priming with long E7 peptide/LPS and IL10 signalling inhibitor, then boosting with long E7 peptide/IFA elicits may lead to better HPV infection related tumour regression in clinic.

Balloon catheter dilation technology combined with a fibrolaryngoscope to treat a maxillary sinus cyst.

A prospective randomized controlled study was conducted to investigate the effect of balloon catheter dilation technology combined with a fibrolaryngoscope in the treatment of a maxillary sinus cyst. The clinical data of 14 cases (19 maxillary sinuses) with balloon catheter dilation technology combined with a fibrolaryngoscope to remove sinus cysts (balloon group) and 16 cases (23 maxillary sinuses) with conventional nasal endoscopic sinus surgery to remove sinus cysts (conventional group) were analyzed. All cases have completed the preoperative and postoperative SNOT-20, nasal endoscopy and coronal sinus CT scan. Lund-Kennedy endoscopic and Lund-Mackay CT scan staging scores were recorded. All patients were followed up for 24 weeks after the operation. The SNOT-20 scores, Lund-Kennedy endoscopic and Lund-Mackay CT scan staging scores were lower in the balloon group than that in the control group. Balloon catheter dilation technology combined with a fibrolaryngoscope can effectively preserve the function and structures of the nasal cavity and sinus, making it a good choice in the treatment of a retention cyst of the maxillary sinus.

Manipulating IL-10 signalling blockade for better immunotherapy.

Interleukin 10 is a cytokine with the ability to reduce or terminate inflammation. Chronic viral infection, such as infection of chronic hepatitis B, hepatitis C and HIV, has increased levels of interleukin 10 in peripheral blood. Serum IL-10 levels are also high in certain cancers. Blocking IL-10 signalling at the time of immunisation clears chronic viral infection and prevents tumour growth in animal models. We review recent advances in this area, with the emphasis on potential use of this novel strategy to treat chronic viral infection and cancer in human.

Applications of titanium mesh tubing in external ear canal reconstruction in congenital aural atresia.

The aim of this study was to observe the effect of a titanium tube on external auditory canal reconstruction in congenital aural atresia and to assess the tube's effectiveness in preventing external canal stenosis or atresia after reconstruction. Reconstruction of the external ear canal with a titanium mesh tube was performed in 16 patients (16 ears) with congenital aural atresia at the First People's Hospital of Foshan. The titanium mesh tube was removed 1 year after surgery. The patients were followed up for 2 years (2 ± 0.3 years), and all of the patients had formed a new external ear canal. There was no local infection, granulation tissue, re-stenosis, or atresia in any of the patients after surgery. All of the patients were content with their newly formed external ear canal. Titanium mesh tubing is safe and effective for reconstruction of the external ear canal during surgery for congenital aural atresia.

Prognostic value of grading masticator space involvement in nasopharyngeal carcinoma according to MR imaging findings.

PURPOSE: To derive a suitable method for grading masticator space invasion in nasopharyngeal carcinoma on the basis of magnetic resonance (MR) images and to determine its prognostic value in patients undergoing intensity-modulated radiation therapy. MATERIALS AND METHODS: After institutional review board approval and informed consent were acquired, 808 patients with nasopharyngeal carcinoma who were treated with definitive intensity-modulated radiation therapy were analyzed retrospectively. The anatomic sites of masticator space involvement were identified with MR imaging. Overall survival, local relapse-free survival, and distant metastasis-free survival were calculated by using the Kaplan-Meier method and were compared by using the log-rank test. Potential prognostic factors were identified by means of multivariate analysis. RESULTS: Masticator space involvement was diagnosed in 163 of 808 patients (20.2%). Patients with lateral invasion (involvement of the lateral pterygoid muscle of the masticator space and beyond) had significantly poorer overall survival and distant metastasis-free survival than those with medial invasion (involvement of the medial pterygoid muscle of the masticator space) (P = .035 and P = .026, respectively). Furthermore, their overall survival, local relapse-free survival, and distant metastasis-free survival were significantly poorer compared with patients with stage T2 or T3 disease (all P ≤ .023) but similar to patients with stage T4 disease. The grade of masticator space involvement was an independent prognostic factor for overall survival, local relapse-free survival, and distant metastasis-free survival (all P ≤ .023). CONCLUSION: Masticator space involvement in nasopharyngeal carcinoma should be graded as medial (stage T2 disease) or lateral (stage T4 disease). This can facilitate staging of nasopharyngeal carcinoma and may be a suitable prognostic indicator of survival.

IL-10 signalling blockade at the time of immunization inhibits Human papillomavirus 16 E7 transformed TC-1 tumour cells growth in mice.

IL-10 signalling blockade by intra-peritoneal injection of anti-IL-10 receptor antibodies at the time of immunization enhances vaccine induced CD8+ T cell responses and promotes bacteria, parasitic and viral control. We now show that blockade of IL-10 signalling at the time of immunization enhances vaccine induced antigen specific CD8+ T cell responses to both dominant and subdominant CTL epitopes. Injection of anti-IL-10 receptor antibodies subcutaneous at the time of immunization also enhances CD8+ T cell responses. Furthermore, IL-10 signalling blockade at the time of a Human papillomavirus 16 E7 peptide/LPS immunization, prevents HPV16 E7 transformed TC-1 tumour growth in mice. Immunization in the presence of anti-IL-10R antibodies and Monophosphoryl lipid A, generates antigen specific CD8+ T cell responses similar to immunization with LPS. Our results suggest that immunization and IL-10 signalling blockade may provide a novel way for the development of therapeutic vaccines against cancer.

High-pressure studies of Bi2S3.

The high-pressure structural and vibrational properties of Bi2S3 have been probed up to 65 GPa with a combination of experimental and theoretical methods. The ambient-pressure Pnma structure is found to persist up to 50 GPa; further compression leads to structural disorder. Closer inspection of our structural and Raman spectroscopic results reveals notable compressibility changes in specific structural parameters of the Pnma phase beyond 4-6 GPa. By taking the available literature into account, we speculate that a second-order isostructural transition is realized near that pressure, originating probably from a topological modification of the Bi2S3 electronic structure near that pressure. Finally, the Bi(3+) lone-electron pair (LEP) stereochemical activity decreases against pressure increase; an utter vanishing, however, is not expected until 1 Mbar. This persistence of the Bi(3+) LEP activity in Bi2S3 can explain the absence of any structural transitions toward higher crystalline symmetries in the investigated pressure range.

Endoscope-assisted second branchial cleft cyst resection via an incision along skin line on lateral neck.

The aim of the study is to report the feasibility of endoscope-assisted second branchial cleft cyst resection via a small incision along the skin line on the lateral neck. In total, 41 patients from the Department of Otolaryngology, Foshan Hospital of Yat-sen University were randomly assigned to conventional (20 patients) or endoscope-assisted (21 patients) second branchial cleft cyst resection. The patient clinical characteristics, operation time, operative bleeding volume, postoperative complications, and subjective satisfaction with the incision scar (measured using a visual analog scale) were compared between the groups. All 41 s branchial cleft cyst resections were successfully performed, and the wounds healed uneventfully. The bleeding volume (6.3 ± 2.5 ml) and incision length (2.7 ± 0.3 cm) differed between the groups (P < 0.00). The mean patient satisfaction score was 8.0 ± 0.8 in the endoscope-assisted surgery group and 6.4 ± 0.9 in the control group (P < 0.00). All of the patients in the endoscope-assisted surgery group were satisfied with their cosmetic results. No marginal nerve palsy occurred. No complications such as bleeding, salivary fistula, or paresis of the marginal mandibular branch occurred. All of the patients were disease free through a follow-up period of 6-24 months (median: 14 months). Endoscope-assisted second branchial cleft cyst resection via a small incision along the dermatoglyph on the lateral neck is a feasible technique. This procedure may serve as an alternative approach, allowing a minimally invasive incision and better cosmetic results.

Modified endoscope-assisted partial-superficial parotidectomy through a retroauricular incision.

OBJECTIVES: The aim of this study was to evaluate the feasibility of an endoscope-assisted partial parotidectomy through a modified retroauricular incision. PATIENTS AND METHODS: Thirty patients with benign parotid superficial lobe tumors with a diameter of 2.4 ± 0.5 cm, located in the anterior portion of the inferior auricular lobule, underwent an endoscope-assisted partial-superficial parotidectomy. A retrograde approach through a small skin incision was used. An additional 30 patients who underwent conventional surgeries were used as controls. The operation time, operative bleeding volume and subjective satisfaction with the incision scar were compared between the groups. RESULTS: All operations were successfully performed. The endoscopic surgery duration (74.8 ± 15.7 min), bleeding volume (12.7 ± 3.9 ml) and incision length (4.8 ± 0.4 cm) differed between the groups (p = 0.001). The mean patient satisfaction score was 8.6 ± 1.2 in the endoscope-assisted surgery group and 5.4 ± 1.3 in the control group (p = 0.001). There were no tumor recurrences during the 9-36 months of follow-up. CONCLUSION: Endoscope-assisted partial-superficial parotidectomy via a modified retroauricular incision is a feasible method for the treatment of benign parotid superficial lobe tumors located in the anterior portion of the inferior auricular lobule. The main advantage of this procedure was that the small operative scars improved the cosmetic results.

Pressure-Induced Changes in the Crystal Structure and Electrical Conductivity of GeV4S8.

Lacunar spinels, represented by AM4X8 compounds (A = Ga or Ge; M = V, Mo, Nb, or Ta; X = S or Se), form a unique group of ternary chalcogenide compounds. Among them, GeV4S8 has garnered significant attention due to its distinctive electrical and magnetic properties. While previous research efforts have primarily focused on studying how this material behaves under cooling conditions, pressure is another factor that determines the state and characteristics of solid matter. In this study, we employed a diamond anvil cell in conjunction with high-energy synchrotron X-ray diffraction, Raman spectroscopy, four-point probes, and theoretical computation to thoroughly investigate this material. We found that the structural transformation from cubic to orthorhombic was initiated at 34 GPa and completed at 54 GPa. Through data fitting of volume vs pressure, we determined the bulk moduli to be 105 ± 4 GPa for the cubic phase and 111 ± 12 GPa for the orthorhombic phase. Concurrently, electrical resistance measurements indicated a semiconductor-to-nonmetallic conductor transition at ∼15 GPa. Moreover, we experimentally assessed the band gaps at different pressures to validate the occurrence of the electrical phase transition. We infer that the electrical phase transition correlates with the valence electrons in the V4 cluster rather than the crystal structure transformation. Furthermore, the computational results, electronic density of states, and band structure verified the experimental observation and facilitated the understanding of the mechanism governing the electrical phase transition in GeV4S8.

The low binding affinity of ADAMTS4 for citrullinated fibronectin may contribute to the destruction of joint cartilage in rheumatoid arthritis.

OBJECTIVES: Rapid cartilage degradation in the joints is observed in rheumatoid arthritis (RA). ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs-4) degrades aggrecan, the primary component of cartilage, therefore contributing to joint erosion in RA. The proteolytic activity of ADAMTS4 is inhibited by fibronectin (FN). FN is abundantly expressed in the synovia in RA and is modified by citrullination, the conversion of peptidylarginine to citrulline. This study aims to investigate the binding ability of citrullinated FN (cFN) to ADAMTS4 and the effect of cFN on aggrecanase activity. METHODS: The full-length recombinant ADAMTS4 was purified from HEK293 cells that were transiently transfected with a full-length cDNA coding for human ADAMTS4. A 40-kDa FN fragment exhibiting heparin binding was citrullinised with rabbit peptidylarginine deaminase. The binding activity of the full-length recombinant ADAMTS4 to cFN was investigated in an in vitro binding assay. The proteolytic activity of ADAMTS4 after incubation with cFN was determined using an aggrecanase activity kit, in which the ARGSVIL peptide is produced by digestion with aggrecanase. RESULTS: cFN displayed significantly decreased binding activity with ADAMTS4 compared with FN. The full-length ADAMTS4 produced large amounts of the ARGSVIL peptide, but the amount was markedly decreased in the presence of FN. The production of this peptide approached the normal level when the full-length ADAMTS4 was incubated with cFN. CONCLUSIONS: FN following citrullination is less effective in inhibiting the proteolytic activity of ADAMTS4. It is known that PADI4, an enzyme active in citrullination, is highly expressed in the synovial tissue in RA. Our results suggest that PADI4 in the RA synovium may contribute to cartilage destruction via the citrullination of FN.

Vitamin D-binding protein (group-specific component) has decreased expression in rheumatoid arthritis.

OBJECTIVES: This study used a proteomic approach to screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles to that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients. The result was complemented by a SNP analysis. METHODS: Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis. The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF/TOF MS. The result was verified using western blotting. Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA, 51 patients with AS and 160 healthy controls. The genotyping result was confirmed in a large cohort of 389 patients with RA, 200 patients with AS and 371 healthy controls. RESULTS: The proteomic approach detected significantly decreased expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA, which was confirmed with western blot analysis. rs2282679 was significantly associated with RA and AS (p=0.026794 and 0.007566, respectively). The result was confirmed in a large cohort of RA (OR=0.678639, 95%CI=[0.541113~0.851118], p=0.000776) and AS (OR=0.564053, 95%CI=[0.433716~0.733558], p=1.79e-005). CONCLUSIONS: 1,25-dihydroxyvitamin D3 inhibits cell proliferation, immunoglobulin production and the release of cytokines through binding to VDBP. VDBP also mediates bone resorption by activating osteoclasts. The decreased expression and the genetic effect of VDBP in RA suggest a novel pathogenic pathway that vitamin D contributes to the arthritic process of the disease.

The microRNA miR-29c-5p inhibits cell proliferation and migration by targeting TMEM98 in head and neck carcinoma.

Head and neck squamous cell carcinoma (HNSCC), which occurs frequently worldwide, is characterized by high risk of metastasis. MicroRNAs (miRNAs) play crucial roles in tumorigenesis and cancer development. In this study, miR-29c-5p was identified using three high throughput microarrays. We measure miR-29c-5p expression in HNSCC tissues and cell lines. To determine the function of miR-29c-5p in HNSCC, we evaluated its effects in vitro on cell proliferation, the cell cycle, apoptosis, and cell migration. We employed a mouse tumor xenograft model to determine the effects of miR-29c-5p on tumors generated by HNSCC cell lines. The miR-29c-5p expression was lower in HNSCC tissues than in normal tissues. Upregulated miR-29c-5p expression in HNSCC cells inhibited migration and arrested cells in the G2/M phase of the cell cycle. Further, upregulated miR-29c-5p expression inhibited the proliferation of HNSCC cells in vivo and in vitro. In addition, transmembrane protein 98 (TMEM98) was identified as a direct target of miR-29c-5p by using a luciferase reporter assay. These findings provide new insights that link the regulation of miR-29c-5p expression to the malignant phenotype of HNSCC and suggest that employing miR-29c-5p may serve as a therapeutic strategy for managing patients with HNSCC.

UBR5 regulates proliferation and radiosensitivity in human laryngeal carcinoma via the p38/MAPK signaling pathway.

Laryngeal carcinoma (LCC) is a common malignant tumor with low radiosensitivity and generally poor response rates. The ubiquitin protein ligase E3 component n­recognin 5 (UBR5) has prognostic implications in several neoplasms; however, its role in LCC and radiotherapy sensitivity remains unknown. Immunohistochemistry and bioinformatics analyses were performed to measure UBR5 protein and mRNA expression in LCC and adjacent non­tumor tissues. The gene and protein expression of UBR5 in LCC and HuLa­PC cell lines were measured using quantitative PCR and western blot analyses. Following transfection with small interfering RNA or UBR5 overexpression plasmid in LCC cells, the proliferation, cell cycle distribution, invasion, migration and radiosensitivity of LCC cells were analyzed. UBR5­related lncRNA, targeted miRNA and protein­protein interaction networks were analyzed using bioinformatics. Finally, the expression of the p38/mitogen­activated protein kinase (MAPK) pathway was evaluated following UBR5 silencing in M2E cells treated with radiation. Increased UBR5 expression was observed in LCC tissues compared with adjacent non­tumor tissues, and it was correlated with poor overall survival of LCC patients. After overexpression or silencing of UBR5 in M2E and M4E LCC cells, cell proliferation and radiosensitivity were significantly increased or decreased, respectively, compared with the control groups. The percentage of S phase cells decreased in the UBR5 si­RNA group compared with that in the control group, while overexpression of UBR5 exerted no effect on the cell cycle. In addition, the expression of Bcl­2 and p38 was decreased in the si­UBR5 combined with radiation groups. The level of phosphorylated p38 expression was increased after combination of si­UBR5 with radiation. The small molecule inhibitor of p38/MAPK signaling, SB203580, decreased the viability of UBR5­overexpressing cells and the survival fraction when cells were exposed to radiation. These findings demonstrated that UBR5 may be involved in regulating cell proliferation and sensitivity to radiotherapy in LCC via the p38/MAPK pathway, thereby highlighting its possible value for the development of new therapeutic strategies and targets for the treatment of this disease.

Topical application of temperature-sensitive caerin 1.1 and 1.9 gel inhibits TC-1 tumor growth in mice.

Genital warts, which are one of the most common sexually transmitted diseases (STDs), result from persistent infection with human papillomavirus (HPV), especially subtypes 6 or 11. Topical application of 5% imiquimod cream is currently recommended as a first-line treatment choice for genital warts, but the clearance and patient compliance rates remain less than sufficient. In the current study, we developed a temperature-sensitive gel that contains the host-defense peptides caerin 1.1 and 1.9, which were originally isolated from Australian tree frogs of the genus Litoria. Growth of HPV16 E6/E7-transformed TC-1 cells was inhibited in vitro and in vivo following injection of the tumor with the caerin gel in a TC-1 tumor mouse model. Furthermore, when the caerin gel was topically applied, the inhibitory effect remained, and T, NK cells were attracted to the tumor site. In addition, the gel maintained a similar level of bioactivity after incubation at room temperature for 30 days. Our results suggest that this caerin gel, following further optimization, may provide an alternative method for the management of genital warts.

Host-Defense Peptides Caerin 1.1 and 1.9 Stimulate TNF-Alpha-Dependent Apoptotic Signals in Human Cervical Cancer HeLa Cells.

Host defense caerin 1.1 and 1.9 peptides, isolated from the glandular secretion of Australian tree frogs, the genus Litoria, have been previously shown to have multiple biological activities, including the inhibition of human papillomavirus (HPV) 16 early protein E7 transformed murine as well as human cancerous cell proliferation both in vitro and in vivo. However, the mechanism underlying their anti-proliferative activities against HPV18+ cervical cancer HeLa cells remains unknown. This study comparatively investigated the anti-proliferation on HeLa cells by caerin 1.1, 1.9, and their mixture, followed by confocal microscopy examination to assess the cellular intake of the peptides. Tandem mass tag labeling proteomics was employed to reveal the proteins that were significantly regulated by the peptide treatment in cells and cell growth environment, to elucidate the signaling pathways that were modulated. Western blot was performed to confirm the modulation of the pathways. Both caerin 1.1 and 1.9 highly inhibited HeLa cell proliferation with a significant additive effect compared to untreated and control peptide. They entered the cells with different magnitudes. Intensive protein-protein interaction was detected among significantly upregulated proteins. Translation, folding and localization of proteins and RNA processing, apoptosis process was significantly enriched post the treatments. The apoptotic signaling was suggested as a result of tumor necrosis factor-α (TNF-α) pathway activation, indicated by the dose-dependent elevated levels of caspase 3 and caspase 9. The epidermal growth factor receptor and androgen receptor pathways appeared inhibited by the peptides. Moreover, the activation of T-cell receptor derived from the quantitation results further implies the likelihood of recruiting more T cells to the cell growth environment post the treatment and more sensitive to T cell mediated killing of HeLa cells. Our results indicate that caerin 1.1 and 1.9 mediate apoptotic signals of HeLa cells and may subsequently enhances adaptive T cell immune responses.