[Predictive value of D-dimer for deep venous thrombosis of lower extremity in adult burn patients].

Objective: To investigate the predictive value of D-dimer for deep venous thrombosis (DVT) of lower extremity in adult burn patients. Methods: A retrospective case series study was conducted. The clinical data of 3 861 adult burn patients who met the inclusion criteria and were admitted to the Department of Burns of Zhengzhou First People's Hospital from January 1, 2015 to December 31, 2019 were collected. The patients were divided into DVT group (n=77) and non-DVT group (n=3 784) according to whether DVT of lower extremity occurred during hospitalization or not. Data of patients in the two groups were collected and compared, including the gender, age, total burn area, D-dimer level, with lower limb burn and inhalation injury or not on admission, with sepsis/septic shock, femoral vein indwelling central venous catheter (CVC), history of surgery, and infusion of concentrated red blood cells or not during hospitalization. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, and chi-square test. The indicators with statistically significant differences between the two groups were analyzed with multivariate logistic regression analysis to screen the independent risk factors for DVT of lower extremity in 3 861 adult burn patients. The receiver operating characteristic (ROC) curve of the independent risk factors predicting DVT of lower extremity in 3 861 adult burn patients were drawn, and the area under the curve (AUC), the optimal threshold value, and the sensitivity and specificity under the optimal threshold value were calculated. The quality of the AUC was compared by Delong test, and the sensitivity and specificity under the optimal threshold value were compared using chi-square test. Results: There were no statistically significant differences in gender, occurrence of sepsis/septic shock or history of surgery during hospitalization between patients in the two groups (P>0.05), while there were statistically significant differences in age, total burn area, D-dimer level, lower limb burn and inhalation injury on admission, and femoral vein indwelling CVC and infusion of concentrated red blood cells during hospitalization between patients in the two groups (t=-8.17, with Z values of -5.04 and -10.83, respectively, χ2 values of 21.83, 5.37, 7.75, and 4.52, respectively, P<0.05 or P<0.01). Multivariate logistic regression analysis showed that age, total burn area, and D-dimer level were the independent risk factors for DVT of lower extremity in 3 861 adult burn patients (with odds ratios of 1.05, 1.02, and 1.14, respectively, 95% confidence intervals of 1.04-1.06, 1.00-1.03, and 1.10-1.20, respectively, P<0.05 or P<0.01). The AUCs of ROC of age, total burn area, and D-dimer level for predicting DVT of lower extremity in 3 861 adult burn patients were 0.74, 0.67, and 0.86, respectively (with 95% confidence intervals of 0.68-0.80, 0.60-0.74, and 0.83-0.89, respectively, P values<0.01), the optimal threshold values were 50.5 years old, 10.5% total body surface area, and 1.845 mg/L, respectively, the sensitivity under the optimal threshold values were 71.4%, 70.1%, and 87.0%, respectively, and the specificity under the optimal threshold values were 66.8%, 67.2%, and 72.9%, respectively. The AUC quality and sensitivity and specificity under the optimal threshold value of D-dimer level were significantly better than those of age (z=3.29, with χ2 values of 284.91 and 34.25, respectively, P<0.01) and total burn area (z=4.98, with χ2 values of 326.79 and 29.88, respectively, P<0.01), while the AUC quality and sensitivity and specificity under the optimal threshold values were similar between age and total burn area (P>0.05). Conclusions: D-dimer level is an independent risk factor for DVT of lower extremity in adult burn patients, its AUC quality and sensitivity and specificity under the optimal threshold value are better than those of age and total burn area, and it has good predictive value for DVT of lower extremity in adult burn patients.

[Efficacy and safety of giant emphysematous bulla volume reduction via medical thoracoscope].

Objective: To evaluate the efficacy and safety of giant emphysematous bulla (GEB) volume reduction via medical thoracoscope. Methods: This was a prospective, single-arm study conducted between July 2018 and September 2020 in Ri Zhao Hospital of Traditional Chinese Medicine. Patients who met the inclusion criteria were treated with GEB volume reduction via medical thoracoscope and were followed up to evaluate the efficacy and safety of the technique. According to comparison of preoperative and postoperative chest CT results, the self-designed evaluation criteria of imaging efficacy were as follows: complete or nearly complete disappearance of GEB (GEB volume reduction ≥90%), significant reduction of GEB (75%≤GEB volume reduction<90%), reduction of GEB (50%≤GEB volume reduction<75%) and no change (GEB volume reduction<50%). Results: A total of 47 patients were included, among whom 43 were males, with an age M (Q1, Q3) of 63.0 (55.0, 67.0). The CT results showed complete or nearly complete disappearance of GEB in 43 patients, significant reduction of GEB in 3 patients and reduction of GEB in 1 patient before discharge. The degree of dyspnea improved significantly (P<0.05). Arterial partial pressure of carbon dioxide (PaCO2) decreased from (48.2±8.4)mmHg (1 mmHg=0.133 kPa) to (45.4±7.3)mmHg (P<0.05). The 6-minute walk test (6MWT) increased from (245.6±162.4)m to (283.5±152.2)m (P<0.05). Six-month postoperative follow-up was completed in 24 patients, and CT results showed that the efficacy of volume reduction was continuous compared with that before discharge. GEB was further reduced or even disappeared in 3 of the cases. Besides, the degree of dyspnea, 6MWT (384.4±148.2)m and PaCO2 (42.7±6.6)mmHg were improved significantly (P<0.05). The oxygenation index (356.86±61.21)mmHg was significantly higher than that before surgery (295.20±67.16)mmHg and before discharge (294.50±76.69)mmHg (P<0.05). No perioperative deaths occurred. Conclusions: GEB volume can be completely eliminated or significantly reduced by this innovative technique, while PaCO2, the degree of dyspnea and exercise endurance can be significantly improved after operation. The 6-month follow-up after surgery showed that the above benefits continued, and that the oxygenation index improved significantly.

Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay.

OBJECTIVE: To evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and compare it with RT-PCR. METHODS: We designed primers specific to the orf1ab and S genes of SARS-CoV-2. Total viral RNA was extracted using the QIAamp Viral RNA Mini Kit. We optimized the RT-LAMP assay, and evaluated it for its sensitivity and specificity of detection using real-time turbidity monitoring and visual observation. RESULTS: The primer sets orf1ab-4 and S-123 amplified the genes in the shortest times, the mean (±SD) times were 18 ± 1.32 min and 20 ± 1.80 min, respectively, and 63°C was the optimum reaction temperature. The sensitivities were 2 × 101 copies and 2 × 102 copies per reaction with primer sets orf1ab-4 and S-123, respectively. This assay showed no cross-reactivity with 60 other respiratory pathogens. To describe the availability of this method in clinical diagnosis, we collected 130 specimens from patients with clinically suspected SARS-CoV-2 infection. Among them, 58 were confirmed to be positive and 72 were negative by RT-LAMP. The sensitivity was 100% (95% CI 92.3%-100%), specificity 100% (95% CI 93.7%-100%). This assay detected SARS-CoV-2 in a mean (±SD) time of 26.28 ± 4.48 min and the results can be identified with visual observation. CONCLUSION: These results demonstrate that we developed a rapid, simple, specific and sensitive RT-LAMP assay for SARS-CoV-2 detection among clinical samples. It will be a powerful tool for SARS-CoV-2 identification, and for monitoring suspected patients, close contacts and high-risk groups.

[Clinical analysis between trefoil factor 3 gene polymorphsims and susceptibility to papillary thyroid carcinoma].

Objective:To explore the relationships between trefoil factor 3（TFF3） gene polymorphisms and susceptibility to papillary thyroid carcinoma（PTC） in Han population of northern China. Method:A case-control study was performed in 123 PTC patients and 108 healthy controls. Four SNPs in the TFF3 gene, including rs225361, rs533093, rs9981660 and rs225439, were detected by gene sequencing. Result:Compared with healthy people, there was no significant difference in the genotype frequencies of rs225361, rs9981660, rs533093 and rs225439 alleles in the PTC group（P>0.05）. The CGTC and CGTT haploids of TFF3 gene were positively correlated with the occurrence of PTC, and CGCC and TGTC haploids were negatively correlated with the occurrence of PTC. TT genotype of rs9981660 had significant differences in the distribution of PTC with and without lymph node metastasis（P<0.05）. Conclusion:Polymorphisms in 4 SNP loci in the TFF3 gene may be unrelated to the occurrence of PTC. The CGTC, CGTT, CGCC and TGTC haploids in the TFF3 gene might be related to the development of PTC. The TT genotype at rs9981660 may be associated with lymph node metastasis of PTC.

[Diagnostic performance of ultrasound-based risk stratification systems for indeterminate thyroid nodules: comparison of the 2015ATA guidelines with the 2016 KTA/KSThR and 2017ACR guidelines].

Objective: To evaluate the diagnostic value of the 2015ATA, 2017ACR and 2016KTA/KSThR ultrasound model for indeterminate thyroids nodules. Method: Four hundred and sixty-four patients with thyroid nodules that were initially diagnosed as indeterminate thyroids nodules by fine needle aspiration(FNA) cytology were included in this study. The clinical data and two-dimensional ultrasonographic features were compared between the benign and malignant nodules. The two-dimensional ultrasound images of all nodules were sorted by the 2015ATA, 2017ACR and 2016KTA/KSThR guideline ultrasound model grading criteria, and the malignant risk of different grading were calculated. In order to calculate the diagnosis and other indicators, the optimal threshold drawing from ROC curve was drawn to obtain the cut-off value of 2015ATA, 2017ACR and 2016KTA/KSThR. Result: ①There was no significant difference in age, sex and nodule size between benign and malignant nodules(P>0.05), and there also was no significant difference in irregular margin, microcalcification between benign and malignant nodules.②The sensitivity of the 2015ATA ultrasound model was 87.9%, slightly lower than that of the 2016KTA/KSThR and 2017ACR guidelines(P>0.05).The specificity of 2015ATA was 63.9%, which was significantly higher than that of 2016KTA/KSThR and 2017ACR guidelines(P<0.05). There was no significant difference between 2015ATA ultrasound model and 2016KTA/KSThR guide ultrasound for the accuracy(P>0.05), but the accuracy of 2015ATA ultrasound model was significantly higher than that of 2017ACR guide(P<0.05).③The area under the curve of 2015ATA was slightly lower than that of 2016KTA/KSThR(0.889 VS 0.902, P>0.05) and significantly higher than that of 2017ACR(0.889 VS 0.854, P<0.05). Conclusion: 2015ATA has high specificity and accuracy and moderate sensitivity for the diagnosis of benign and malignant indeterminate thyroids nodules, which is helpful for the clinical evaluation and management of such nodules.

[The role of SWE and ATA(2015) guidelines combined mode in differentiation malignant from benign of Bethesda â ¢ thyroid nodules].

Objective: To evaluate the diagnostic value of the ATA (2015) ultrasound model, shear wave elastography (SWE), and ATA (2015)+SWE combinative modality for the diagnostic efficiency in thyroid nodules with Bethesda Classification Ⅲ indeterminate cytology, determine the diagnostic value of ultrasonography in Bethesda Classification Ⅲ indeterminate cytology.Method:216 thyroid nodules that were initially diagnosed as AUS/FLUS by fine needle aspiration (FNA) were included in this study. The clinical data and two-dimensional ultrasonographic features were compared between the benign and malignant nodules. The two-dimensional ultrasound images of all nodules were sorted by the 2015ATA guideline ultrasound model grading criteria. The maximum and average shear wave velocity (SWV) values were obtained from multiple SWV measurement under the VTIQ speed mode. The optimal threshold drawing from ROC curve and diagnostic performance of single and combinative modality were calculated.Result:①There was no significant difference in age, sex and nodule size between benign and malignant nodules (P>0.05). Malignant nodules of 152 cases of AUS thyroid nodules had significantly higher rates of not well-circumscribed margin and presence of microcalcifications (P=0.005,P=0.004). ②There were significant differences in malignancy risk among the different US patterns defined by the 2015 ATA guidelines in AUS nodules. ③The maximum and mean SWV of AUS/FLUS nodules measured in VTIQ mode were statistically significant in evaluating benign and malignant nodules. ④The area under ROC curves of ATA (2015)+SWE combined mode was 0.912, larger than single diagnosis mode ［ATA (2015):0.854, SWE: 0.862］.Conclusion:SWE can not only compensate for the deficiency of ATA (2015) in the diagnosis of benign and malignant FLUS thyroid nodules, but also effectively improve the diagnostic performance of ATA (2015) in the differentiation of benign and malignant AUS thyroid nodules.

[Efficacy and safety of argon plasma coagulation plus medical adhesive via thoracoscopy in the treatment of spontaneous pneumothorax with subpleural blebs].

Objective: To evaluate the preliminary efficacy and safety of argon plasma coagulation (APC) plus medical adhesive via thoracoscopy in the treatment of spontaneous pneumothorax caused by subpleural blebs. Methods: Data of totally 51 patients of spontaneous pneumothorax with subpleural blebs treated in Pneumology Department of Rizhao Hospital of Traditional Chinese Medicine from July 2015 to July 2017 were retrospectively analyzed. Among the 51 patients, 59 target bullae were found, 1.2 per patient on average, with the diameter ranging from 0.5 to 5 cm and an average diameter of 2.6 cm. A total of 21 patients were treated with APC, and the rest 30 were treated with APC plus partially sprayed medical adhesive (APC+ group). The rate of air leakage discontinuance within 24 hours after operation, the rate of air leakage discontinuance within one week after operation, the disappearance rate of target subpleural blebs on CT scans one week after operation, and the rate of significantly shrunken target subpleural blebs and wall thickness, as well as the incidence of postoperative complications, including fever, chest pain, pleural effusion, hemorrhage, and infection after operation were observed and compared between the two groups. Results: The air leakage discontinuance rate in APC+ group was significantly higher than that in APC group 24 hours after operation (90.0% vs 52.4%, P<0.05), and the rate in APC+ group was also significantly higher than that in APC group one week after operation (96.7% vs 66.8%, P<0.05). There was no significant difference in the disappearance rate of target subpleural blebs on CT scans one week after operation and the incidence of significantly shrunken target subpleural blebs and wall thickness (both P>0.05). There was no significant difference in the incidence of postoperative complications such as fever, chest pain and pleural effusion (all P>0.05). Conclusion: The treatment of spontaneous pneumothorax with subpleural blebs by APC plus medical adhesive is safe and effective.

[The effects of PM 2.5 and air quality index on the emergency of otolaryngology].

Objective:To discuss the effects of PM 2.5 and air quality index on the emergency of otolaryngology. Method:Collect in 2015 Beijing tongren hospital emergency department patients with acute otitis media, epistaxis, acute epiglottitis as well as the same time Beijing AQI value and the PM 2.5 density data as the experimental group 1, chooses in 2015 first Affiliated Hospital of Hebei North University similar patient as well as the same time Zhangjiakou AQI value and the PM 2.5 density as the experimental group 2. Statistics the quantity of each sickness of two groups, and the relationships with PM 2.5 and AQI value; Group all data by AQI value and statistics the differences in the incidence rates of each group. Result:The number of cases of acute otitis media and epistaxis in the 2 experimental group were all correlated with the change of PM 2.5 and AQI value, and the correlation was positive between the two groups, However, the incidence of acute epiglottitis was not significantly correlated with the concentration of PM 2.5 and AQI, and there was no significant correlation between the two. The air quality when severe and more pollution when the number of the cases of acute otitis media and epistaxis compared with good air quality for the number of P<0.05, the difference was statistically significant. Conclusion:The increase of PM 2.5 concentration and AQI index is a health risk factor for acute otitis media and epistaxis.

Lentivirus-mediated shRNA interference of trefoil factor 3 blocks cell viability, migration and invasion in the papillary thyroid carcinoma cells.

Trefoil factor 3 (TFF3), a regulatory protein composed of 59 amino acids, has been suggested to be involved in pathogen- esis, proliferation, invasion, migration and apoptosis in multiple malignant tumors. However, the roles of TFF3 concerning the viability, migration and invasion in papillary thyroid carcinoma cells have not yet been studied. This study aimed to investigate the effect of TFF3 knockdown on a thyroid papillary carcinoma TPC-1 cell line both in vitro and in vivo. In the present study, lentivirus-mediated short hairpin RNA (shRNA) targeting TFF3 plasmids were first constructed and stable TPC-1 cells were obtained while their TFF3 gene was silenced with either shTFF3-TPC-1, or a scrambled shRNA control. TFF3 expression was detected using quantitative real-time PCR and western blot analyses. The TPC-1 cell viability was measured by CCK-8 assay and colony formation. The cell migration and invasion were assessed by wound scratch assay and transwell filters. AKT phosphorylation, MMP-9, and BCL-2 expression levels were detected by western blot analyses. Our results showed that TFF3 knockdown significantly inhibits TPC-1 cell viability, migration and invasion. AKT phosphoryla- tion, MMP-9, and BCL-2 levels were all remarkably depressed in TFF3 knockdown TPC-1 cells. Using a thyroid papillary carcinoma xenograft mouse model, we further investigated the effects of TFF3 knockdown in vivo. Significantly delayed xenograft emerging, slower growth rate and lower final tumor weights and volumes were observed in the shTFF3 group as compared to the control group. As expected, the expression levels of MMP-9 and BCL-2 in the xenograft are consistent with those of shTFF3-TPC-1 and shTFF3-TPC-1 cells in vitro. Our results suggest that TFF3 plays a vital role in the viability and oncogenesis of TPC-1 cells and may be a potential target for effective treatment of thyroid papillary carcinoma.

[Assess the diagnostic value of MicroPure system with ultrasonography combined with mSMI in diagnosing thyroid nodules in TI-RADS 3 and 4].

Objective:To assess the diagnostic value of MicroPure system with ultrasonography combined with mSMI in diagnosing thyroid nodules inTI-RADS3 and 4; to compare the testing capability in thyroid nodules micro salcifications between MicroPure system and grey-scale ultrasonography; and to compare the diagnostic efficiency between mSMI and color doppler flow imaging (CDFI).Method:Exerting gery-scale ultrasonography and CDFI to choose 86 patients with 106 nodules (64 TI-RADS3 class nodules; 42 TI-RADS4 class nodules). These thyroid nodules were detected by grey-scale ultrasonography, CDFI, MicroPure system with ultrasonography, mSMI, and they were graded by TI-RADS. Then the ultrasound diagnosis value were compared with their FNA and surgical pathology results by gery-scale ultrasonography and CDFI; grey-scale ultrasonography and CDFI and MicroPure system; grey-scale ultrasonography and mSMI and MicroPure system, and diagnostic efficiency between mSMI and CDFI were compared, and the detectability of thyroid nodules micro salcifications between MicroPure system and grey-scale ultrasonography.Result:The results showed that combination of MicroPure system and mSMI had a higher sensitivity (91.44%, P< 0.05); The MicroPure system showed a higher testing capability compare to grey-scale ultrasonography in thyroid nodules micro salcifications (P< 0.05); The mSMI showed a higher sensitivity compared to the CDFI (P< 0.05).Conclusion:Combining mSMI and MicroPure system could improve the discreminability of TI-RADS3, 4 class thyroid nodules, then provide evidence to assess,diagnosis and treat the patients in clinical.

[The role of ATA (2015) guidelines, superb microvascular imaging, and spectral Doppler in differentiation between malignant and benign thyroid nodules].

Objective:To evaluate the diagnostic value of ultrasound patterns of 2015 American Thyroid Association (ATA) guidelines, color Doppler flow imaging (CDFI), superb microvascular imaging (SMI), and spectral Doppler in the differentiation of benign and malignant thyroid nodules.Method:The study included 254 thyroid nodules confirmed by fine needle aspiration cytology (FNAC) or histopathologic examination. All nodules were detected by ultrasonography, SMI, CDFI and spectral Doppler respectively. Blood flow detection technologies (CDFI and SMI) were used to classify flow distribution characteristics of thyroid nodules according to Kim's grading criteria. The optimal threshold drawing from ROC curve is calculated to obtain the cut-off value of spectral Doppler.Result:①For benign solid nodules with peripheral flow distribution and malignant solid nodules with central blood flow distribution, the display rates of SMI are higher than those of the CDFI, the differences were statistically significant (P< 0.05). ②There was a significance difference in detecting benign and malignant nodules using spectral Doppler (P< 0.05). ③The specificity and accuracy of combinative modality 2015 ATA+SMI was significantly higher than that of ATA, SMI, and spectral Doppler. There was no significant difference between ATA+SMI and ATA+SMI+spectral Doppler for the sensitivity, specificity, and accuracy.Conclusion:SMI had a better detection rate of blood vessels than CDFI and contribute a certain value in differential diagnosis of benign and malignant thyroid nodules. The combination of 2015 ATA and SMI can improve diagnostic value in the differentiation between benign and malignant thyroid nodules.

[Expression and clinical significance of Nrf2 and ERK pathway in papillary thyroid carcinoma].

Objective:To explore the expression and clinical significance of nuclear factor erythroid-2 related factor 2 (Nrf2) and ERK sinal in papillary thyroid carcinoma (PTC). Method:PTC microassy were detected by immunohistochemical method (IHC). The expression of Nrf2, ERK2 and p-ERK½ in PTC, para-carcimoma and normal thyroid tissue were detected by Western blot. The transcriptional level of Nrf2 mRNA in PTC, para-carcimoma and normal thyroid tissue were detected by real-time quantitative PCR (qPCR) method. Result:Nrf2, ERK2, p-ERK½ protein were high expression in group of lymph node metastasis PTC and negative or weak positive expression in para-carcinoma tissue (P<0.01). And the proteins of clinical stage Ⅲ-Ⅳ of PTC groups express higher than without lymph node metastasis group and clinical stage Ⅰ-Ⅱ of PTC group (P<0.05). The expression of Nrf2 protein was positively correlated with ERK2 and p-ERK½ protein in PTC (r=0.723 2,r=0.668 6,P<0.05). The Western blot results of Nrf2, ERK2, p-ERK½ had a higher level in PTC than those of paracarcinoma and normal thyroid tissue (P<0.01). The level of Nrf2 mRNA were higher than those of paracarcinoma and normal thyroid tissue (P<0.01). Conclusion:High expresion of Nrf2 may be associated with the progression of papillary thyroid carcinoma, and may relate to the high expression and activity of ERK pathway.

[Study on anti-oxidase in smoking-related laryngeal squamous cell carcinoma].

Objective:To investigate the role of oxidative stress in smoking-related laryngeal squamous carcinoma through detecting the expression of antioxidant enzymes in smoking patients. Method:A total of 138 cases with laryngeal squamous cell carcinoma enrolled in the first hospital affiliated the northern he bei college from 2012 to 2015 and forty five volunteers were conducted. All participants were divided into three groups according to smoking index: group A(heavy smoking, 88 cases of laryngeal cancer patients) and group B(no smoking 50 cases of laryngeal cancer patients) and C group(45 heavy smoking volunteers).Catalase(CAT), glutathione peroxidase(GSH-px) and malondialdehyde(MDA) and the expression of NRF2 in serum, tissue adjacent to carcinoma, and carcinoma tissues from each groups were measured, respectively. Result:①the expression of the CAT and GSH-px in group A were significantly lower than that of group B(P <0.05), but higher than that of group C(P <0.05); ②the MDA level of group A is significantly higher than group B(P <0.05) and C group(P <0.01);③NRF2 was highly expressed in carcinoma tissues, and the expression level was negatively correlated with degree of carcinoma differentiation (P <0.05). Conclusion:Compared with nonsmoking patients, heavy smoking patients with laryngeal cancer were under more severe oxidative stress. NRF2 expression level in patients with laryngeal squamous cell carcinomas was associated with pathological stage.

[Investigation of CXCR4 mediated chemoresistance in nasopharyngeal carcinoma cell line CNE2].

Objective:Since nasopharyngeal carcinoma is easy to develop resistance during cisplatin-based chemotherapy,CXCR4 expression levels were elevated in mang tumors,and the factor to do with tumor metastasis and chemotherapy drug resistance,and so on has a very important link.We established cisplatin-resistant nasopharyngeal carcinoma cell line, named as CNE2/DDP, and investigated the function of CXCR4 in molecular mechanism behind this resistance.Method:CNE2/DDP was firstly build up by increasing concentration of cisplatin. And then afterwards,MTT assay, RNA interference techniques, microRNA overexpresion techniques, quantative PCR and western blotting were applied to analyze the function of CXCR4 and its downstream effectors.Result:①the expression of CXCR4 was increased in CNE2/DDP and downregulation of CXCR4 with CXCR4 siRNA was able to decrease the resistance of CNE/DDP to cisplatin; ②the expression of let-7a was decrease in CNE2/DDP, while the expression of bcl-2 was increased. Upregulation of let-7a via transfection of let-7a mimics could downregulate the expression of bcl-2 and damage the resistance of CNE2/DDP to cisplation;③downregulation of CXCR4 through CXCR4 siRNA transfection was capable of improving the expression of let-7a. Conclusion:We were the first to found that CXCR4 was related to chemoresistance of CNE2/DPP to cisplatin. Meanwhile, we confirmed that CXCR4 affected the expression of bcl-2 through regulating the expression of let-7a to modulate the chemoresistance of CNE2/DPP to cisplatin.

The status of PD-L1 and tumor-infiltrating immune cells predict resistance and poor prognosis in BRAFi-treated melanoma patients harboring mutant BRAFV600.

BACKGROUND: BRAF inhibitors (BRAFi) improve survival in metastatic melanoma patients (MMP) but the duration of clinical benefit is limited by development of drug resistance. Here, we investigated whether the expression of programmed death-ligand 1 (PD-L1) and the density of tumor-infiltrating mononuclear cells (TIMC) predict the occurrence of resistance, hence affecting the clinical outcome in BRAFi-treated MMP. METHODS: PD-L1 expression (cutoff 5%) was analyzed by immunohistochemistry with two different antibodies in BRAF(V600)-mutated formalin-fixed and paraffin-embedded samples from 80 consecutive MMP treated with BRAFi at a single institution. TIMC were evaluated by conventional hematoxylin and eosin staining. RESULTS: Forty-six and 34 patients received vemurafenib and dabrafenib, respectively. Membranous expression of PD-L1 was detected in 28/80 (35%) of patients. At multivariate analysis, absence of tumoral PD-L1 staining [odd ratio (OR) 10.8, 95% confidence interval (CI) 2.7-43.3, P < 0.001] and the presence of TIMC (OR 6.5, 95% CI 1.7-24.3, P < 0.005) were associated with a better response to treatment. Median progression-free survival (PFS) and overall survival were 10 and 15 months, respectively. By multivariate assessment, PD-L1 expression [hazard ratio (HR) 4.3, 95% CI 2.1-8.7, P < 0.0001] and absence of TIMC (HR 2.5, 95% CI 1.4-4.7, P < 0.002) correlated with shorter PFS. PD-L1 overexpression (HR 6.2, 95% CI 2.8-14.2, P < 0.0001) and absence of TIMC (HR 3.1, 95% CI 1.5-6.5, P < 0.002) were independent prognostic factors for melanoma-specific survival. CONCLUSION: Our results provide the first proof-of-principle evidence for the predictive and prognostic relevance of PD-L1 immunohistochemical expression and density of immune cell infiltration in BRAF(V600)-mutated MMP treated with BRAFi.

c-Myc-mediated repression of miR-15-16 in hypoxia is induced by increased HIF-2α and promotes tumor angiogenesis and metastasis by upregulating FGF2.

Previous studies have established the link between aberrant microRNA (miRNA) expression and hypoxia in various neoplasms. However, how these hypoxia-related miRNAs modulate tumor progression is still unclear. Therefore, the patterns of miRNA in colorectal carcinoma cell lines in response to hypoxia or not were first screened and the hypoxia-induced repression of the miR-15-16 cluster was confirmed. Then, this repression was found to be associated with high tumor stage and poor prognosis in colorectal carcinoma and is shown to promote tumor angiogenesis and metastasis by the loss of restriction of its target gene, fibroblast growth factor-2 (FGF2). Moreover, the general and alterative promoters of the miR-15-16 host (deleted in lymphocytic leukemia 2, DLEU2) were mapped, and three c-Myc/Max binding sites in response to the hypoxia-induced repression of miR-15-16 were further identified. Finally, an enhanced stability of c-Myc/Max heterodimer promoted by increased hypoxia-inducible factor-2α (HIF-2α) was validated, and we also verified that the enhancement contributed to the hypoxia-induced repression of miR-15-16. In brief, the c-Myc-mediated transcriptional repression of miR-15-16 in hypoxia is induced by increased HIF-2α and promoted tumor angiogenesis and hematogenous metastasis by the further loss of post-transcriptional inhibition of FGF2. Our study provides a better understanding of the coping mechanisms in response to tumor hypoxia and may be helpful in developing an effective prognostic marker or treatment target against solid tumors.

Expression and localization of trefoil factor family genes in rat submandibular glands.

The trefoil factor (TFF) family, which comprises TFF1, TFF2 and TFF3, plays an essential role in epithelial regeneration within the gastrointestinal tract. All three TFFs are present in human saliva; TFF3 is the predominant trefoil peptide. Little is known about the expression and tissue distribution of TFFs in rats, which are commonly used as a model system for human studies. We investigated the localization of the TFF genes that encode secretory peptides in rat submandibular glands (SMG). All three TFFs were expressed in rat SMG, although their location varied. Substantial amounts of TFF1 were detected only in the cytoplasm of epithelial cells in the SMG granular convoluted tubules (GCT), while TFF2 and TFF3 were widely distributed in the cytoplasm of epithelial cells of intercalated ducts (ID), striated ducts (SD) and interlobular ducts (ILD). The three TFFs also were detected especially in the lumens of the SD and ILD. Semi-quantitative RT-PCR and in situ hybridization experiments confirmed TFF1, TFF2 and TFF3 mRNA expressions in the SMG. Greater expression of TFF peptides and mRNA was observed in male rats than in females. The broad expression of TFFs in rat SMG cells and lumens suggests that TFFs function in this organ by their secretion into the duct lumens. We also found differences in TFF expression profiles between rat and human SMG; therefore, caution should be exercised when using rats as a model for human TFF studies.

Mer receptor tyrosine kinase promotes invasion and survival in glioblastoma multiforme.

The infiltration of glioma cells into adjacent tissue is one of the major obstacles in the therapeutic management of malignant brain tumours, in most cases precluding complete surgical resection. Consequently, malignant glioma patients almost invariably experience tumour recurrences. Within the brain, glioma cells migrate rapidly either amoeboidly or mesenchymally to invade surrounding structures, in dependence on the extracellular environment. In addition, radiotherapy, frequently applied as adjuvant therapeutic modality, may enhance tumour cell mobility. Here, we show that the receptor tyrosine kinase Mer (MerTK) is overexpressed in glioblastoma multiforme (GBM) and that this is accompanied with increased invasive potential. MerTK expression is maintained in primary GBM-derived tumour spheres under stem cell culture conditions but diminishes significantly in serum-containing cultures with concomitant downregulation of Nestin and Sox2. Depletion of MerTK disrupts the rounded morphology of glioma cells and decreases their invasive capacity. Furthermore, the expression and phosphorylation of myosin light chain 2 are strongly associated with MerTK activity, indicating that the effect of MerTK on glioma cell invasion is mediated by actomyosin contractility. Finally, DNA damage robustly triggers the upregulation and phosphorylation of MerTK, which protects cells from apoptosis. This effect is strongly impaired upon MerTK depletion or overexpression of an inactive MerTK mutant. Collectively, our data suggests that MerTK is a novel therapeutic target in the treatment of the malignant gliomas.

PEG-coated irinotecan cationic liposomes improve the therapeutic efficacy of breast cancer in animals.

BACKGROUND: Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death among females owing. AIM: This study aimed to construct a kind of PEG-coated irinotecan cationic liposomes for investigating its efficacy and mechanism of action in the treatment of breast cancer in preclinical models. MATERIALS AND METHODS: Evaluations were performed on the MDA-MB231 breast cancer cells, the xenografted MDA-MB231 cancer cells in Female nude mice and Sprague-Dawley (SD) rat. The liposomes were characterized through assays of cytotoxicity, intracellular uptake, nuclei morphology, antitumor activities, pharmacokinetics and tissue distribution. RESULTS: The zeta potential of PEG-coated irinotecan cationic liposomes was approximately 23 mV. The PEG-coated irinotecan cationic liposomes were approximately 66nm in diameter, significantly increased the intracellular uptake of irinotecan, and showed strong inhibitory effect on MDA-MB231 breast cancer cells. A significant antitumor efficacy in the xenografted MDA-MB231 breast cancer cells in nude mice was evidenced by intravenous administration of PEG-coated irinotecan cationic liposomes. PEG-coated irinotecan cationic liposomes also improved the irinotecan blood circulation time and showed an enhanced drug concentration in tumor. CONCLUSIONS: PEG-coated irinotecan cationic liposomes had significant inhibitory effect against breast cancer in vitro and in vivo, hence providing a new strategy for treating breast cancer.