[The role and mechanism of human trophoblastic cell surface antigen 2 in the invasion and metastasis of ovarian cancer].

Objective: To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer. Methods: Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, ß-catenin, caspase3, bcl-2, E-cadherin and vimentin. Results: Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells (P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference (P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group (P<0.05). The proportion of G(0)/G(1) cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G(0)/G(1) arrest was observed in SKOV3-shRNA cells (P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group (P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and ß-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions: Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/ß-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.

Establishment of an animal model of spontaneous cervical lymph node metastasis of laryngeal squamous cell carcinoma and obtaining laryngocarcinoma cells with high metastatic potential.

To establish an animal model of spontaneous cervical lymph node metastasis of laryngeal squamous cell carcinoma and obtain laryngocarcinoma cells with high metastatic potential, laryngeal squamous cell carcinoma cell line HEP-2 in logarithmic phase were inoculated under the lingual margin mucosa of nude mice. HEP-2 cells metastasized to the cervical lymph nodes were isolated, cultured, and re-inoculated under the lingual margin mucosa of nude mice twice. The tumor formation in the tongue and in the cervical lymph nodes was confirmed by pathological examination. Carcinoma cells' ability of invasion and migration was detected by transwell assay. Human specific Alu sequences were detected by PCR, which indicated that the tumor cells originated from human laryngeal squamous cell carcinoma cell line HEP-2. Finally, an animal model of spontaneous lymph node metastasis of laryngeal squamous cell carcinoma was successfully established. Laryngeal squamous cell carcinoma cells with high metastatic potential to lymph nodes were obtained through repeated inoculations. .

[Reconstruction of complex tissue defects in temporal region: report of 3 cases].

Objective: To discuss the techniques and repairing methods of various degree of compound tissue defects in the auriculotemporal region. Methods: Retrospective analysis was conducted on three cases of different repairing methods for huge compound tissue defects in different degrees in the auriculotemporal region after the resection of the malignant tumor or sinus tract due to repeated infection in our hospital. Results: Following total removal of the tumors or sinus tract in all patients, we applied retroauricular lingual flap transfer repairing, latissimus dorsi flap free transfer repairing and vascular anastomosis, scalp tissue expansion in stage Ⅰ, then repairing the lesion with expanded scalp and filling the huge mastoid cavity with abdominal fat in stage Ⅱ, respectively, according to the characteristics of compound tissue defects in the auriculotemporal region. All free flaps survived well. Conclusions: The anatomy of the auricular-temporal area is complex and involves important vascular and neural structures of head and neck and lateral skull base. The huge composite tissue defect following auriculotemporal region surgery, which is composed of skin, muscle and bone tissue, needs to be repaired in one stage. Therefore, flexible repairing methods should be chosen based on different situations, for attaining the goal of completely removing tumor and lesions, and then, covering the operation cavity.

[A comparison of the 7th and 8th edition of the AJCC TNM staging systems for predicting survival in oropharyngeal cancer].

Objective:The aim of this study is to evaluate the prognostic value of the 8th edition of AJCC TNM classification in comparison with the 7th edition for oropharyngeal cancer. Method: The clinical and pathological data of 142 patients with oropharyngeal squamous cell carcinoma were retrospectively analyzed. The p16 protein was detected by immunohistochemical staining. Survival and survival analysis were performed using the Kaplan-Meier method and the Log-rank test. Result:The 5-year disease-specific survival（DSS）,the 5-year overall survival (OS) and the median survival time for patients with oropharyngeal cancer were 55.4%,50.4% and 60.3 months, respectively. P16-immunostaining was scored positive in 39 samples (27.5%,39/142).Based on the seventh edition of AJCC staging, there were 4 patients in stage Ⅰ,17 patients in stage Ⅱ,27 patients in stage Ⅲ and 94 patients in stage Ⅳ,respectively.However, there was no significant difference in survival between the patients with stage Ⅰ-Ⅱ and those with Ⅲ-Ⅳ(5-year DSS:75.9% of stage Ⅰ-Ⅱ patients vs. 52.2% of stage Ⅲ-Ⅳ patients;P=0.109).In comparison, 22 patients were diagnosed as stage Ⅰ,24 were stage Ⅱ,27 were stage Ⅲ and 69 were stage Ⅳ using the eighth edition. The 5-year DSS for patients with stage Ⅰ,Ⅱ,Ⅲ and Ⅳ was 88.4%, 80.5%, 73.2% and 29.1%, respectively. The overall difference was statistically significant (P=0.000). Conclusion:Compared with the seventh edition staging, the eighth edition of the AJCC oropharyngeal cancer staging system considers the prognosis of patients with different causes of oropharyngeal cancer, and can more accurately predict the prognosis of patients.

[Clinical study of postoperative adjuvant radiotherapy and postoperative concurrent chemoradiotherapy for locally advanced hypopharyngeal squamous cell carcinoma].

Objective: Using propensity score matching method (PSM) to investigate the clinical effect of postoperative adjuvant radiotherapy and postoperative concurrent chemoradiotherapy for locally advanced hypopharyngeal squamous cell carcinoma. Methods: From July 2007 to July 2018,174 postoperative patients with locally advanced hypopharyngeal squamous cell carcinoma were enrolled in pre-PSM cohort, including 168 males and 6 females, the median age was 60 years old (ranged from 37 to 79 years old).Loco-regional control (LRC),progression-free survival (PFS) and overall survival (OS) were compared and analyzed between the patients treated with postoperative adjuvant radiotherapy and postoperative concurrent chemoradiotherapy (cisplatin was given in a dose of 80 mg/m(2) on days 1, 22, and 43). After the propensity score matching (PSM), 61 sub-pairs of 122 patients were generated in post-PSM cohort. Survival rate were assessed with Kaplan-Meier method and Log-rank test. Results: After the propensity score matching(PSM), 61 sub-pairs of 122 patients were generated in post-PSM cohort.The patients were followed up for 3-135 months, the median follow-up was 42 months. No significant differences in 3-year and 5-year LRC, PFS, OS were observed between the two groups (P>0.05) . For postoperative patients who had high-risk factors (extracapsular extension of nodal disease, and/or vascular embolism, and/or lymph node metastasis≥2, and/or positive surgical margin, and/or perineural infiltration),there were significant differences between the two groups in 3-year PFS (60.99% vs 84.49%,P<0.05), 5-year PFS (35.47% vs 56.97%,P<0.05) and 5-year LRC (41.02% vs 68.50%, P<0.05), but no significant difference was found in OS between the two groups (P>0.05). Conclusion: Postoperative concurrent chemoradiotherapy was more efficacious than postoperative radiotherapy alone in terms of loco-regional control and PFS for high-risk postoperative patients with locally advanced hypopharyngeal squamous cell carcinoma.

Cost-effectiveness analysis of oral probiotics for the prevention of Clostridium difficile-associated diarrhoea in children and adolescents.

BACKGROUND: The incidence of Clostridium difficile-associated diarrhoea (CDAD) in hospitalized children and adolescents has been increasing year-on-year. Paediatric CDAD places a significant economic burden on healthcare systems. Probiotics are live organisms thought to improve the microbial balance of the host, counteract disturbances in intestinal flora, and reduce the risk of colonization by pathogenic bacteria. AIM: A cost-effectiveness analysis was conducted to assess the economy of probiotics for the prevention of CDAD in children and adolescents receiving antibiotics. METHODS: A decision tree model combining clinical effectiveness, utility and cost data was used. Sensitivity analyses were conducted to determine the robustness of the model outcomes. FINDING: The 'oral probiotics' strategy and 'no probiotics' strategy offered patients 0.05876 and 0.056 quality-adjusted life years (QALYs) at a cost of $16,668.70 and $20,355.28, respectively. The oral probiotics strategy exhibited higher QALY and lower cost, and represents the cost-saving strategy. The results were robust for sensitivity analyses. CONCLUSION: From the perspective of the medical system, oral probiotics as a preventive strategy for CDAD in hospitalized children and adolescents receiving a therapeutic course of antibiotics reduced the risk of CDAD and represents a cost-saving strategy.

[A retrospective study on combined modality therapy with or without surgery for advanced hypopharyngeal squamous cell carcinoma: an analysis of 119 cases].

Objective: To compare the treatment outcomes for locally advanced hypopharyngeal carcinoma between surgery plus radio(chemo) therapy(SRT) and non-surgery chemoradiotherapy(CRT). Methods: A total of 119 patients diagnosed with advanced hypopharyngeal carcinoma without distant metastases between 2010 and 2014 were identified in the Chinese People's Liberation Army General Hospital, and they were divided into 2 groups: 42 cases in SRT group and 77 cases in CRT group. Patients' clinical information was collected. Survival rates and prognostic factors were analyzed by the Kaplan-Meier method with SPSS 23.0 software. The survival rates, laryngeal preservation rates and complication rates were compared between the two groups using the chi-square test.Among the 119 patients, 112 were males and 7 were females. Age ranged from 27 to 78 years, with an average age of 57 years. Results: There were no significant difference between the SRT and CRT group for five-year disease-free survival (DFS, 53.9% vs. 45.1%, χ(2)=1.251, P=0.263) and overall survival (OS, 54.9% vs. 45.6%, χ(2)=1.749, P=0.186). Compared to SRT group, CRT group did not showed the significant increase of treatment complications (χ(2)=0.858, P=0.354), with a higher laryngeal preservation rate (50.0% vs. 71.4%, χ(2)=6.493, P=0.011). Conclusions: Advanced hypopharyngeal carcinoma is of high malignancy and poor prognosis. Combined modality treatment is a main approach for advanced hypopharyngeal cancer. SRT offers disease-free survival and overall survival rates equivalent to CRT, but with a higher laryngeal preservation rate.

[Clinical features and prognostic factors of squamous cell carcinoma of the base of tongue：an analysis of 124 cases].

Objective:To summarize the clinicopathological characteristics and prognostic factors of squamous cell carcinoma (SCC) of the base of tongue (BOT). Method:Review the clinical data of 124 cases of patients with BOT tumors. HPV-DNA was detected using SPF10-DNA enzyme immunoassay and LiPA genotyping method. Survival rates and prognostic factors was calculated with SPSS 19.0 software using the Kaplan-Meier method and multivariate Cox model survival analysis. Result:5-year disease-specific survival (DSS), 5-year overall survival and median survival time for patients with SCC BOT were 54.5%, 48.5% and 45.2 months, respectively. HPV infection rate was 18.3%(11/60), the HPV-16 subtype in 10 cases, accounting for 90.9%(10/11). Univariate analysis results showed that smoking history (P=0.028), drinking history (P=0.001), tumor stage (P=0.009) and p16 protein expression (P=0.015) significantly affected the prognosis. The prognosis of patients with HPV positive BOT cancers was better than the HPV negative patients (5-year DSS was 80.8% compared to 54.5%), but not statistical difference (P=0.138). Multivariate analysis revealed that independent prognostic factors of BOT tumors were drinking history (P=0.006) and tumor stage (P=0.035). Conclusion:The prognosis of BOT tumor was relatively poor. Smoking history, drinking history, tumor staging and the expression of p16 protein significantly affected the prognosis. Drinking history and tumor stage were independent prognostic factors for BOT cancers.

[Thyroid adenoma in the piriform fossa: a case report and literature review].

Objective: To explore the clinical characteristics of case of thyroid adenoma in the piriform fossa, and review the literatures of the congentital thyroid gland abnormality. Methods: A 44-year-old male had foreign body feeling in his pharynx for 3 years. A mass in his left piriform fossa was detected by the clinical and imaging examination. Biopsy report that the mass was thyroid papillary carcinoma. The resection of tumor with partial back thyroid cartilage through lateral neck and pharyngeal approach was performed. Results: The surgical wound healed in first-stage and no any surgical complication occurred. With postoperative pathological and immunohistochemical examination, the mass was finally diagnosed as thyroid gland adenoma. Staining for cytokerantin19 was negative. Conclusion: The symptomatic and neoplastic abnormal thyroid gland should be excised, but asymptomtic, non-neoplastic and functional abnormal thyroid gland should be retained with regular follow up.

[Clinical features of patients with head and neck malignant tumor concurrent with multiple primary carcinoma].

OBJECTIVE: To analyze the clinical characteristics of multiple primary cancers (MPC) associated with head and neck. METHODS: A total of 71 patients with MPC associated with head and neck treated from January 2008 to November 2015 were included in the retrospective study, of them 20 patients were synchronous MPC and 51 patients were metachronous MPC; 45 (63.4%) patients were field cancerization of head neck (FCHN) and 26 (36.6%) patients were non-field cancerization of head neck (NFCHN). For the synchronous MPC patients with an interval of 0 month, the lesion sites of FCHN were treated with combined chemotherapy and radiotherapy, the site of severe lesion of NFCHN received firstly a standard treatment. Other types of MPC were treated according to international guide lines or experts consensus. RESULTS: With the mean follow-up of 63.4 months, the 3-year and 5-year overall survival (OS) were 84.6% and 75.7% respectively. The 3-year and 5-year OS were 56.4% and 37.6% respectively in patients with synchronous MPC, and were 92.2% and 84.2% respectively in patients with metachronous MPC. There was significant difference in the OS between patients with metachronous MPC and patients with synchronous MPC (P=0.0002). The 3-year and 5-year OS were 85.7% and 77.9% respectively in patients with FCHN, and were 82.9% and 72.8% respectively in patients with NFCHN, with no significant difference between the two groups (P=0.297). CONCLUSION: With correct diagnosis and effective curative treatment, some of patients with MPC can get long-term survival, showing the better prognosis in metachronous MPC compared to synchronous MPC.

Renal abnormalities in schoolchildren.

To establish prevalence of some renal abnormalities in schoolchildren, an epidemiologic study of 132,686 schoolchildren, including 69,903 boys and 62,783 girls, was conducted from March 1987 to May 1988 in the city of Taipei. At the health station of each school, the students were screened quickly by a physician with portable real-time ultrasound equipment. When a renal abnormality was detected or suspected, the examinee was referred to Taipei Municipal Women and Children's Hospital for further investigation. Radiologic and urologic procedures were then selectively performed to establish the correct diagnosis. Renal abnormalities were detected in 645 students (approximately 0.5% of total population screened). There were 256 cases of hydronephrosis, 103 cases of unilateral renal agenesis, 128 cases of unilateral small kidney, 90 cases of renal cystic disorders, 30 cases of ectopic kidney, and 38 cases of other abnormalities. Surgically correctable lesions were demonstrated in 50 of these students. Rapid renal ultrasonography was found to effectively detect some renal abnormalities initially, and prevalence could then be established after further investigations.

Mutagenicity of benzidine and related compounds employed in the detection of hemoglobin.

Seven compounds commonly used as chromagens for the detection of hemoglobin and its derivatives have been assayed for mutagenicity employing the Salmonella/mammalian microsome test. Three of these compounds, benzidine, o-dianisidine, and o-tolidine, were shown to be mutagenic. Since benzidine and o-tolidine are already known to be carcinogens, there is a high probability that o-dianisidine will also prove to be a carcinogen. Four compounds tested with this system, o-anisidine, diphenylamine, guaicol, and o-toluidine, were not mutagenic.

A rapid method to produce anti-gentamicin antibody.

The aminoglycoside antibiotic, gentamicin, was conjugated to erythrocytes or bovine serum albumin (BSA) by a simple procedure in which ECDI was employed as the coupling reagent. When rabbits were immunized by injecting gentamicin-goat erythrocyte conjugates, three kinds of antibody were produced: 1. anti-gentamicin antibody, 2. anti-ECDI antibody, 3. goat erythrocyte agglutinins. The interfering anti-ECDI antibody was easily neutralized by adding acidified ECDI solution to the immune serum. Goat agglutinins were avoided by employing rabbit erythrocytes as the carrier cell in the hemagglutination titration. Highly specific anti-gentamicin antiserum was produced in rabbits by first injecting an initial dose of gentamicin-BSA conjugate as an emulsion in incomplete Freund's adjuvant via the foot pad, followed by multiple intravenous injections of gentamicin-erythrocyte conjugates. The immunization took approximately 21 days. High titered anti-gentamicin antibody was also produced by foot pad inoculation of gentamicin-BSA conjugates; however, the time necessary to achieve comparable titers was considerably longer (55 days). The antibodies produced by both immunization procedures were mainly of the IgG class.

[The state of cytoskeleton and its links "oxygen-peroxide" effects in some pathologies and apoptosis].

The cytoskeleton elements, especially the system of the microtubules, are responsible for production of cell backbone system, creation the global spatial ordered organization for efficient transport processes. Microtubules are involved into transportation of mitochondria, peroxisomes, microsomes, lyzosomes, Golgi apparatus, vesicular structures, some enzymes, adhesion molecules and, possibly, O2-depoting compounds. During disorganization of microtubules, mitochondria (principal consumers of intracellular O2), lose uninterrupted "address" delivery oxidizing substrates and O2. This may be one of important factors underlying mitochondrial dysfunctions accompanied by a rise and/or intensification of cells hyperoxia and oxidative stress. These impairments are obviously responsible for oxygen-peroxide effects in aging, age-related pathologies, carcinogenesis and apoptosis. The agents, both stabilizing and disorganizating microtubules appear to be apoptogenic for the tumors cells.

Autoantibody against beta1-adrenergic receptor and left ventricular remodeling changes in response to metoprolol treatment.

Autoantibodies specific for the beta(1)-adrenoceptor (beta(1)-AR) have been implicated in the pathology of congestive heart failure (CHF). We hypothesized that the presence of autoantibodies against beta(1)-AR (anti-beta(1)-AR) is associated with left ventricular (LV) remodelling in response to metoprolol. Synthetic beta(1)-AR peptides served as the target antigen in an ELISA (enzyme-linked immunosorbent assay) were used to screen the sera of 106 CHF patients. Patients were separated into positive (+) anti-beta(1)-AR or negative (-) anti-beta(1)-AR groups according to their anti-beta(1)-AR reactivity. Echocardiography (ECG) was performed at baseline and after one year of metoprolol therapy in combination with standard treatment regime for CHF, that is, digoxin, diuretics and an ACEI (angiotensin-converting enzyme inhibitor). The dose of metoprolol was doubled on a biweekly basis up to 50 mg x 2 daily (b.i.d./day) or attainment of maximum tolerated dose. Ninety-six patients completed final data analysis. Fifty-four patients with (+) anti-beta(1)-AR had greater improvements than 42 patients with (-) anti-beta(1)-AR in LVEDD (left ventricular end-diastolic dimension) (P < 0.01, from 69 +/- 0.8 to 58.0 +/- 0.5 mm vs. 69.0 +/- 0.8-63.6 +/- 0.9 mm) and LVESD (left ventricular end-systolic dimension) (P < 0.01, from 57.1 +/- 1.4 to 43.9 +/- 0.8 mm vs. 56.2 +/- 0.9-48.6 +/- 1.0 mm), and LVEF (left ventricular ejection fraction) (P < 0.01, from 35.4 +/- 1.3 to 49.8 +/- 0.6% vs. 34.4 +/- 1.0-44.3 +/- 1.1%) by metoprolol therapy in combination with standard treatment regime for one year. Of the CHF patients with (+) anti-beta(1)-AR, 65.4% responded to target metoprolol dose as compared to 21.4% of CHF patients without anti-beta(1)-AR (P < 0.01). Response to target metoprolol dose occurred more rapidly in (+) anti-beta(1)-AR than (-) anti-beta(1)-AR of CHF patients (67.5 +/- 2.4 vs. 100.8 +/- 3.0 days, P < 0.01). These results demonstrated that CHF patients with (+) anti-beta(1)-AR had greater improvements in LV remodelling and heart function by metoprolol as compared to (-) anti-beta(1)-AR patients. Moreover, patients with (+) anti-beta(1)-AR have better tolerance to metoprolol therapy than patients without anti-beta(1)-AR.