The Clinical Significance of Detecting Blood Supply to the Inferior Parathyroid Gland Based on the "Layer of Thymus-Blood Vessel-Inferior Parathyroid Gland" Concept.

Objective: How to preserve the inferior parathyroid gland (IPTG) in situ during central neck dissection (CND) is the major concern of thyroid surgeons. The "layer of thymus-blood vessel-IPTG" (TBP layer) concept showed to be effective in preserving IPTG. The objective of this study was to identify the origin and course of blood supply to IPTG (IPBS) within the TBP layer and to take key points of operation during CND. Design: This is a retrospective control study. Participants. Patients who underwent thyroidectomy plus CND using the TBP layer concept and conventional technique between 2017 and 2019 were enrolled. Measurements. The origin and course of IPBS in relation to recurrent laryngeal nerve (RLN) and thymus and prevalence of hypoparathyroidism were detected. Results: A total of 71.3% of IPTGs (251 of 352) were supplied by ITA branches, defined as type A. Type A was further divided into Types A1 (branches of ITA, coursing laterally to the RLN (53.1%, 187 of 352)) and A2 (branches of ITA, traversing medially to the RLN (18.2%, 64 of 352)). Type A2 was more common on the right side than on the left side (P < 0.001). Fifty-five (15.6%) IPTG feeding vessels originated from the thymus or mediastinum. Nineteen (5.4%) IPTGs were supplied by branches of the superior thyroid artery. The incidence of transient hypoparathyroidism decreased from 45.7% to 3.6% (P < 0.001), in the TBP layer group compared with the conventional technique group. Conclusion: The origin and course of IPBS follow a definite pattern. This mapping and precautions help surgeons optimize intraoperative manipulations for better preservation of IPBS during CND.

A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation.

A novel, simple, and sensitive quantitative polymerase chain reaction (qPCR) technology, which is termed as helper qPCR, was established to detect miRNA. In this assay, the target miRNA sequence was introduced as helper template for a reaction switch preforming two-step real-time qPCR strategy. Firstly, the reverse primer was reverse transcribed to form "mediator primer" after binding to the target miRNA. Then, the mediator primer was further extended to form "active template" with annealing to the mediator template. In the end, the active template was amplified and detected by the qPCR reaction system with the help of reverse and forward primers. The SYBR Green dye was used for fluorescence quantification, which is quicker and cheaper than the fluorescent probes, as the detection limit of this assay was 1 pM. This helper qPCR system can be used for different miRNAs detection by redesigning reverse primer for target, indicating this strategy could afford good performance in detecting multiple miRNAs and has a promising application prospect.

Excellent diagnostic performance of FNA-Tg in detecting lymph nodes metastases from papillary thyroid cancer.

Background: Thyroglobulin washout of fine needle aspiration (FNA-Tg) has proved to be useful in detecting lymph node metastases from papillary thyroid cancer; however, the influences of thyroid gland, Hashimoto thyroiditis, serum thyroglobulin (Tg) and anti-TG antibody on the diagnostic performance of FNA-Tg are controversial. Patients & methods: We retrospectively collected the FNA-Tg results of 176 preoperative or postoperative patients (356 lymph nodes) who finally were diagnosed with papillary thyroid cancer. The diagnostic abilities of FNA-Tg were evaluated and compared under different circumstances. Results: The diagnostic performance of FNA-Tg was uninfluenced irrespective of the status of thyroid gland or serum anti-TG antibody. However, high serum Tg was positively correlated with FNA-Tg (Exp(B) = 1.57; 95% CI: 1.209-2.309; p = 0.001). Conclusion: FNA-Tg was an excellent diagnostic tool, but it should be interpreted with caution only if serum Tg is higher than 10 ng/ml.

Eukaryotic translation initiation factor 5A-2 involves in doxorubicin-induced epithelial-mesenchymal transition in oral squamous cell carcinoma cells.

Background: Epithelial-mesenchymal transition (EMT) is considered to be vital during chemotherapy resistance in oral squamous cell carcinoma (OSCC). Recently, eukaryotic initiation factor 5A-2 (eIF5A-2), a potential oncogene, has been reported to be involved in chemotherapy resistance in human cancers. Materials and Methods: N1-guanyl-1,7-diaminoheptane (GC7, a novel eIF5A-2 inhibitor) or siRNA on responses to doxorubicin were examined in OSCC cells. Cytotoxicity and protein expression were evaluated by CCK-8 and EdU incorporation assay and western blotting. Tca8113 cells were used for establishment and treatment of tumor xenografts in vivo. Results: Low concentration of GC7 (5µΜ) significantly enhanced doxorubicin cytotoxicity in both epithelial phenotype OSCC cells (Cal27) and mesenchymal phenotype OSCC cells (HN30 and Tca8113). EMT process promoted by doxorubicin in Cal27 cells could be reversed by GC7. Additionally, GC7 induced mesenchymal-epithelial transition (MET) in HN30 and Tca8113 cells. Silencing of eIF5A-2 by specific siRNA exhibited the similar effects. The synergistic cytotoxicity of doxorubicin/GC7 combination was not induced in Twist-1, an EMT driving factor, silenced Cal27, HN30, and Tca8113 cells. GC7 also synergized doxorubicin to inhibit tumor growth in vivo treatment. Conclusions: Our study strongly proved that combined treatment with GC7 may boost the therapeutic effect of doxorubicin in OSCC by inhibiting the EMT.

GC7 enhances cisplatin sensitivity via STAT3 signaling pathway inhibition and eIF5A2 inactivation in mesenchymal phenotype oral cancer cells.

Eukaryotic initiation factor 5A2 (eIF5A2), a newly identified oncogene, promotes cell survival, proliferation and motility in tumorigenesis. Drug resistance and dose-related adverse side-effects greatly reduce the efficiency and safety of cisplatin-based chemotherapy in advanced or recurrent oral squamous cell carcinoma (OSCC) patients. The present study investigated the effect of eIF5A2 combined with N1-guanyl-1,7-diaminoheptane (GC7, a novel eIF5A2 inhibitor) or siRNA. We found that low concentrations of GC7 (≤5 µM) had little effect on OSCC cell viability, but significantly enhanced cisplatin cytotoxicity. Compared with cisplatin, GC7/cisplatin had little effect on cisplatin-promoted mesenchymal-epithelial transition in mesenchymal phenotype Tca8113 and HN30 cells, or on cisplatin-induced epithelial-mesenchymal transition (EMT) in epithelial phenotype Cal27 and HN4 cells. Further research revealed that the upregulation of p-STAT3 and c-Myc which was induced by the single treatment with either cisplatin or GC7 was significantly reversed by the GC7/cisplatin combination in mesenchymal phenotype Tca8113 and HN30 cells. In in vivo treatment, we revealed that the GC7/cisplatin combination presented significant tumor volume reduction without distinct body weight loss. In conclusion, our data indicated that eIF5A2 is a potent therapeutic target in OSCC treatment. Our results revealed a novel mechanism by which GC7/cisplatin combination therapy may offer an efficient and safe therapeutic alternative to advanced or recurrent OSCC patients.

Thermochemotherapy of lower lip squamous cell carcinoma without metastases: an experience of 31 cases.

INTRODUCTION: The aim of this study was to evaluate the efficacy, functional and aesthetic results, and safety of a novel treatment, thermochemotherapy, for lower lip squamous cell carcinoma (LLSCC) without metastases. PATIENTS AND METHODS: A combination of local hyperthermia delivered by a 915MHz microwave heating system and the chemotherapy of pingyangmycin (bleomycin A(5) hydrochloride) (PYM) and methotrexate (MTX), was administered to 31 patients of LLSCC twice per week for a period of 4.5-7.5 weeks. Patients with complete response (CR) have been followed up for a full five-year period, whereas partial response (PR) patients were excluded for further analysis. The local control of tumour, functional and cosmetic outcomes, recurrence, regional lymph node and distant metastases, and complications were assessed by clinical and imaging examination. RESULTS: Clinical CR was observed in twenty-nine (93.55%) patients and PR in two (6.45%), the total response rate was 100%, while the adverse effects were extremely minimal and tolerable in all 31 patients including 6 elderly patients with a compromised general condition. All 29 CR, including 8 extensive lesions, achieved excellent cosmetic and functional preservation. During the follow-up period, local relapse was seen in 1 case, 1 patient died, and the remainder obtained a complete remission. CONCLUSION: This clinical study suggests that thermochemotherapy may be a feasible treatment for primary LLSCC without cervical metastases, especially for patients with extensive lesions and a compromised general condition.

Central lymph node metastasis: is it a reliable indicator of lateral node involvement in papillary thyroid carcinoma?

BACKGROUND: The purpose of the present study was to evaluate the reliability of central lymph node metastases in predicting lateral node involvement in papillary thyroid carcinoma (PTC). METHODS: This retrospective study consists of 121 patients with PTC who underwent ipsilateral selective neck dissection. The quantitative relationship between central and lateral lymph node metastases (LNM) was analyzed. RESULTS: A total of 86 patients (71.1%) had pathologically proven LNM; 55 patients had LNM in both the central and lateral compartments, 22 patients had only central LNM, and 9 patients had skip metastasis to lateral nodes. The sensitivity of central node involvement to predict lateral LNM was 85.9%, the specificity was 61.4%. The presence of two or more positive central nodes was associated with a high incidence of lateral LNM (70-93.3%), when compared with none (20.5%) or one (47.4%) positive central node. CONCLUSIONS: The node status of the central compartment is moderately predictive of that of lateral compartment. The presence of two or more positive central nodes may be of value in predicting lateral node involvement.

[Relationship between nuclear factor kappa B signal transduction way and the sensitivity of tongue carcinoma cell to Pingyangmycin chemotherapy].

OBJECTIVE: To examine the relationship between NF-kappaB signal transduction way and the sensitivity of Tca8113 carcinoma cell to Pingyangmycin chemotherapy. METHODS: 2 mg/L antisense p65 oligodeoxynucle-otides (5'-GAACAGTTCGTCCATGGCCG-3') was transfected into Tca8113 cells through Lepofectin vectin, then the cells were treated with 8 mg/L Pingyangmycin. After 3 h and 6 h, the expression of p65 in nucleus was studied with immunohistochemical methods and Western blot analyses. After 48 h, the inhibitory rate of cell growth was detected with MTT assay. RESULTS: The NF-kappaB/p65 signal transduction way in Tca8113 cells was activated after treated with Pingyangmycin. But the transfection of antisense p65 oligodeoxynucle-otides inhibited the activation of the signal transduction, the p65 expression in nucleus was decreased significantly (P < 0.05) at 6 h, and the inhibitory rate of cell growth was increased significantly (P < 0.05) at 48 h. CONCLUSION: The results suggested the chemotherapeutic sensitivity of Tca8113 carcinoma cell to Pingyangmycin was improved by the inhibition of NF-kappaB/p65 signal transduction.

[High expression of p65 transcription factor and its significance in tongue carcinoma].

OBJECTIVE: To examine the correlation between p65 expression and the metastasis and prognosis in tongue carcinoma. METHODS: The immunohistochemical expression of p65 was examined in 50 cases of tongue carcinoma, and the clinical and histopathologic records were retrospectively analyzed. RESULTS: Overexpression of p65 in cytoplasma and nucleus was found in 64% cases of tongue carcinoma. There was significant difference of p65 expression among different TNM stages, and the higher expression rate of p65 was found in higher stages. Overexpression of p65 was also related to lymph node metastasis. Significantly greater increase in expression levels of p65 was found in the metastatic cases (n = 22) compared to the nonmetastatic cases (n = 10). However, there was no significant difference of p65 expression among different histopathologic grading. The 5-year survival rate of p65 positive and negative group was 59.3% and 88.9% respectively, and there was significant difference in Kaplan-Meier survival curve (chi2 = 5.6, P < 0.05). CONCLUSION: The results suggest that the expression of p65 was important to predict the metastasis and prognosis in tongue carcinoma.

[Total thyroidectomy with a modified Miccoli's approach for treatment of Graves' disease--feasibility and its applying techniques].

OBJECTIVE: To assess the feasibility and relevant applying techniques of total thyroidectomy for Graves' disease with a modified Miccoli's approach. METHODS: Forty-two patients with Graves' disease consecutively received the radical operation from June 2002 to December 2004.The modification includes: (1) Incision extending according to the degree of lobe enlargement (3-6 cm, average 4 cm); (2) A space maintain-regulating device was used to change dimensionally the volume of working space (mainly height) when specific manipulation needed; (3) A volume-reducing resection step was performed for the gland with degree III hyperplasia by cutting off the middle-inferior part of the lobe prior to endoscopic lobectomy. The approach was designed to mainly use ultrasonically-activated scalpels, with suction-dissector or others as supplementary instruments. During the operations, a method of "sequenced dissect-coagulate-cut" was employed to directly divide all branches of thyroid vessels without ligation or application of hemoclips. RESULTS: All procedures were completed successfully. None of them were converted to open surgery due to uncontrolled bleeding or severe postoperative hematoma. No severe complications occurred, except 2 cases who suffered from temporary hoarseness. CONCLUSION: Total thyroidectomy for Graves' disease can be safely performed with the modified Miccoli's approach by using ultrasonic scalpel and the space maintain-regulating device. Application of these adaptive reforms can obviously reduce the difficulties in manipulation, and thus, make the usage of this minimally invasive design also clinically possible for even radical treatment of the gland.

[The effect of heat shock on the expression of urokinase-type plasminogen activator in human tongue squamous cell carcinoma cell line (Tca8113)].

OBJECTIVE: The aim of this study was to observe the expression of urokinase-type plasminogen activator (UPA) in human tongue squamous cell carcinoma cell line (Tca8113) after heat shock with different dosage. METHODS: The expression of UPA protein of Tca8113 after different heating temperatures(37 degrees C, 40 degrees C, 43 degrees C and 45 degrees C) treatment was examined by immunohistochemical technique (IH) and flow cytometry (FCM). RESULTS: Compared with 37 degrees C group, the UPA protein expression in 40 degrees C and 43 degrees C groups decreased significantly (P < 0.05); however, the UPA protein expression in 45 degrees C group decreased but no statistical difference was found. CONCLUSION: Hyperthermia could inhibit invasion and metastasis of oral squamous cell carcinoma and could be considered as a safe method in curing the tumor.