Expression and Detection Value of 6 Chlamydia Trachomatis Protein Antibodies in Tubal Factor Infertility / 中山大学学报(医学科学版)

ObjectiveTo further study the pathogenic role of different types of Chlamydia trachomatis （CT） proteins in tubal factor infertility， evaluate the clinical detection value of Chlamydia trachomatis protein antibody in predicting tubal factor infertility. MethodsA total of 58 cases of tubal factor infertility （TFI）， 41 cases of fertile controls （FC） and 18 cases of infertile controls （IFC） were included. For serum detection， first， CT-IgG ELISA kit was used to detect the expression of CT-IgG in serum of three groups of people； then， 6 kinds of Chlamydia trachomatis proteins were expressed and purified in the early stage to establish the antibody test for these proteins， and ELISA detection method was used to detect the expression of their antibodies in the serum of TFI group， FC group and IFC group， respectively； and finally， the antibody OD value of the 6 kinds of Chlamydia trachomatis proteins in the three groups of subjects were statistically described， and CT-IgG was used as the reference standard to draw the receiver operating characteristic curve （ROC curve） of each CT antibody. The Youden Index determines the cutoff value for each antibody. Taking TFI as the reference class， two disordered multiple classification logistic regression models were established with the FC and IFC groups， respectively； and the reference class was used to explore the value of various antibodies and age in predicting TFI， FC and IFC of Chlamydia trachomatis. The back-off method was used to screen the variables. ResultsThe OD value of CT376 antibody in the TFI group was higher than that in the FC group （0.86 vs. 0.60， P=0.026）. The CT376 antibody OD value in the TFI group was higher than that in the IFC group （0.86 vs. 0.64， P=0.026）. The CT443 antibody OD value in the IFC group was higher than that in the TFI group （0.59 vs. 0.34， P=0.036） and higher than that in the FC group （0.59 vs. 0.30， P=0.02）. The multiple classification logistic regression analysis established between TFI and FC showed that CT-IgG ［P<0.001， OR=0.084， 95%CI （0.025， 0.284）］， CT376 antibody ［P=0.068， OR=0.359， 95%CI （0.120， 1.078）］. CT-IgG is an independent risk factor for tubal infertility， and CT376 antibody cannot be an independent risk factor for tubal infertility. The multiple classification logistic regression analysis established between TFI and IFC showed that among infertile patients， CT-IgG ［P<0.05， OR=0.194， 95%CI （0.046， 0.817）］， CT376 antibody ［P<0.05， OR=0.176， 95%CI （0.038， 0.818）］ and CT381 antibody ［P<0.05， OR=0.112， 95%CI （ 0.016， 0.796）］ were independent risk factors for tubal infertility. ConclusionThe expression of CT376 antibody in tubal infertility patients is higher than that in fertile and infertile controls， suggesting that CT-induced tubal factor infertility may be related to CT376. CT-IgG， and CT376 antibodies are meaningful in predicting CT-induced tubal factor infertility.

"Graded early warning system" of RET germline mutation carriers in MEN2A/MEN2B families and total thyroidectomy (report of 7 cases) / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the reasonable time of prophylactic thyroidectomy for RET gene carriers in multiple endocrine neoplasia(MEN) 2A/2B families. Methods: From May 2015 to August 2021, RET gene carriers in MEN2A/MEN2B families were dynamically followed up at the Department of Thyroid Head and Neck Surgery, Beijing Tongren Hospital of Capital Medical University. The high-risk patients were encouraged to undergo prophylacitc total thyroidectomy according to the principle of "graded early warning system", namely the evaluation of gene detection, calcitonin value and ultrasound examination successively. Seven cases underwent the surgery, including 3 males and 4 females, aged from 7 to 29 years. According to the risk stratification listed in the guidelines of the American Thyroid Association in 2015, there were 2 cases of the highest risk, 2 cases of the high risk and 3 cases of the modest risk. Calcitonin index remained within the normal range in 3 cases and elevated in 4 cases before operation. All 7 patients underwent thyroidectomy with lymph node dissection of the level Ⅵ performed in 4 patients. Results: The time from suggestion to operation was 2 to 37 months, with an average of 15.1 months. The 6 patients were medullary thyroid carcinoma and 1 case with C-cell hyperplasia. The follow-up time was 2 to 82 months, with an average of 38.4 months. Postoperative serum calcitonin levels of all cases decreased to normal level, with biochemical cure. There was no sign of recurrence on ultrasound examination. All 7 patients had no serious complications, no obvious thyroid dysfunction. Their height, weight and other indicators of pediatric patients were similar to those of their peers, with normal growth and development. Conclusion: For healthy people with MEN2A/MEN2B family history, prophylactic thyroidectomy can be carried out selectively based on the comprehensive evaluation of "graded early warning system" with strict screening and close monitoring.

Utility of GPR68 and TIL in TPF-induced chemotherapy and prognosis evaluation in middle-advanced hypopharyngeal squamous cell carcinoma / 中华耳鼻咽喉头颈外科杂志

Objective: To evaluate the roles of G Protein-Coupled Receptor 68 (GPR68) and tumor infiltrating lymphocytes (TIL) in TPF-(paclitaxel, cisplatin and 5-fluorouracil) induced chemotherapy for middle-advanced hypopharyngeal squamous cell carcinomas. Methods: A total of 31 patients with middle-advanced hypopharyngeal squamous cell carcinoma before TPF-inducted chemotherapy were enrolled from September 2012 to November 2017 in Beijing Tongren Hospital, Capital Medical University, including 28 males and 3 females, aged 43 to 71 years old. The expression of GPR68 and tumor infiltrating CD4+and CD8+T cells before chemotherapy was detected by immunohistochemical staining, and the relationships between GPR68 expression and clinical features, chemotherapy efficacy and overall survival (OS) were analyzed using t-test. Results: After 3 cycles of chemotherapy, there were 4, 14, 10 and 3 patients respectively with complete response (CR), partial response (PR), stable disease (SD) and progressive disease (PD). The positive rates of GPR68 and CD8 were 25% and 40% respectively in the effective group (CR+PR), while 50% and 15% in the ineffective group (SD+PD), with statistically significant differences between two groups (t=5.17 and 12.86,P<0.001). Linear regression analysis showed that GPR68 was negatively correlated with CD8+T cells (r=-0.64,P<0.001). There was no significant correlation between the CD4 expression and TPF efficacy (P>0.05). The mean OS was 12.5 months in patients with high-expressed GPR68 and 25.0 months in patients with low-expressed GPR68, with a statistically significant difference (P=0.005). And mean OS was 25.0 months in patients with high-expressed CD8 and 14.5 months in low-expressed CD8, with a statistically significant difference (HR=2.58, P=0.019). Cox regression analysis showed that GPR68 and CD8+T cells were significant prognostic factors (OR(95%CI)=3.27(2.46-5.97) and 1.53(0.78-1.82), all P<0.05), while CD4 had no significant effect on prognosis (P>0.05). Conclusion: GPR68 and CD8+T cells are expected to be biomarkers for evaluating the efficacy and prognosis of TPF-induced chemotherapy in patients with middle-advanced hypopharyngeal squamous cell carcinoma.

Expression of CD8+Treg cells in chronic rhinosinusitis and its correlation with eosinophilic infiltration / 中华耳鼻咽喉头颈外科杂志

Objective: To detect the percentages of CD8+Treg cells in the nasal mucosa and peripheral blood of chronic rhinosinusitis (CRS) and to explore their correlation with eosinophilic infiltration. Methods: Thirty-three chronic rhinosinusitis with polyp (CRSwNP), 26 chronic rhinosinusitis without polyp (CRSsNP) and 27 control patients who were collected with the nose mucosal tissue and peripheral blood in the Third Affiliated Hospital of Sun Yat-sen University from March 2017 to October 2018 were selected, including 59 males and 27 females, aging from 18 to 72 years. Hematoxylin and eosin (HE) staining was used to observe the number of eosinophils in the nasal tissues and to classify the CRS into eosinophilic CRS (ECRS) and non-eosinophilic CRS (Non-ECRS). Flow cytometry was used to detect the percentages of CD4+ and CD8+T cells in lymphocytes of nasal mucosa and peripheral blood. The percentages of CD8+Foxp3+Treg cells, CD8+Foxp3-IL-10+Treg cells, CD8+IFN-γ+T cells (Tc1), CD8+IL-4+T cells (Tc2) and CD8+IL-17A+T cells (Tc17) in lymphocytes of nasal mucosa and peripheral blood were also tested. Besides, the percentages of Foxp3+TGF-β+Treg cells and Foxp3+IL-10+Treg cells in CD8+T cells were determined. All data were represented by M (IQR). GraphPad 7.0 and SPSS 16.0 were used for illustration and statistical analysis. Results: The percentage of CD8+T cells (37.75%(17.35%)) was higher than that of CD4+T cells (4.72%(4.29%)) in nasal mucosa (Z=-5.70, P<0.001), while lower (23.60%(9.33%)) than that of CD4+T cells (44.05% (10.93%)) in peripheral blood (t=9.72, P<0.001). CRSwNP patients possessed the highest Tc2 (1.82% (1.22%)) and Tc17 (1.93% (2.32%)) percentages than CRSsNP (Tc2: 0.84% (0.79%); Tc17: 0.54% (1.04%)) and control (Tc2: 1.09% (0.92%); Tc17: 0.47% (0.51%), both P<0.05) patients. While, CRSwNP patients possessed the lowest CD8+Foxp3+Treg cells percentage (0.10% (0.32%)) than CRSsNP (0.43% (1.45%)) and control (0.48% (0.83%), Z value was -2.24, -2.22, respectively, P value was 0.025, 0.027, respectively). The percentages of Foxp3+TGF-β+Treg cells and Foxp3+IL-10+Treg cells of CD8+T cells in nasal mucosa in CRSwNP were also lower than controls (Z value was 1.46, 0.49, respectively, both P=0.001). Moreover, the percentage of CD8+Foxp3-IL-10+Treg cells of CD8+T cells was decreased in nasal mucosa of CRSwNP patients (0.14% (0.28%)) when compared with that of CRSsNP (0.89% (0.81%), Z=0.61, P=0.03). ECRS patients had the lower percentages of CD8+Foxp3+Treg cells (0.07% (0.44%)) and CD8+Foxp3-IL-10+Treg cells (0.13% (0.21%)) than Non-ECRS patients (CD8+Foxp3+Treg cells: 0.53% (0.75%); CD8+Foxp3-IL-10+Treg cells: 0.29% (0.76%), t value was 2.14, 2.78, respectively, both P<0.05). The percentage of CD8+Foxp3+Treg cells and the ratio of CD8+Foxp3-IL-10+T per CD8+T cells were negatively correlated with the percentage of eosinophils in CRS patients(R2 value was 0.56, 0.78, respectively, both P<0.001). There was no significant difference in the distribution of CD8+Fxop3+Treg cells and CD8+Fxop3-IL-10+Treg cells in peripheral blood among different groups. Conclusion: The percentages of CD8+Treg cells decrease in CRSwNP patients, especially in ECRS patients, which are opposite to that of Tc2 and Tc17, and negatively correlate with the eosinophils percentage. This indicates that the decrease in the ratio of CD8+Treg cell may be associated with the immune-imbalance and eosinophilic infiltration in nasal mucosa of CRS patients.

A new classification of maxillary defect and simultaneous accurate reconstruction / 中华耳鼻咽喉头颈外科杂志

Objective: To select the preferred flaps for the reconstruction of different maxillary defects and to propose a new classification of maxillary defects. Methods: A total of 219 patients (136 males and 83 females) underwent the simultaneous reconstruction of maxillary defects in the Beijing Tongren Hospital, Capital Medical University, between January 2005 and December 2018 were reviewed. Age ranged from 16 to 78 years. Based on the proposed new classification of the maxillary defects, 22 patients with class Ⅰ defects (inferior maxillectomy), 44 patients with class Ⅱ defects (supperior maxillectomy), 132 patients with class Ⅲ defects (total maxillectomy) and 21 patients with class Ⅳ defects (extensive maxillectomy) were enrolled. Survival rate, functional and aesthetic outcomes of flaps were evaluated. Survival analysis was performed in 169 patients with malignant tumor, Kaplan-Meier method was used to calculate the survival rate, and Log-rank method was used to compare the difference of survival rate in each group. Results: A total of 234 repairs for maxillary defects were performed in 219 patients. Fibula flaps were used in 4/13 of class Ⅰ defects; temporal muscle flaps (11/24, 45.8%) and anterolateral thigh flaps (6/24, 25.0%) used in class Ⅱ defects; temporal muscle flaps (71/128, 55.5%), anterolateral thigh flaps (6/24, 25.0%) and fibula flaps (12/128, 9.4%) used in class Ⅲ defects; and anterolateral thigh flaps (8/20, 40.0%) and rectus abdominis flaps (8/20, 40.0%) used in class Ⅳ defects. The success rate of local pedicled flaps was 95.6% (109/114) and that of free flaps was 95.8% (115/120). Thrombosis(10/234,4.3%) was a main reason for repair failure. Among the followed-up 88 patients, swallowing and speech functions recovered, 82 (93.2%) of them were satisfied with appearance, and 75 (85.2%) were satisfied with visual field. The 3-year and 5-year overall survival rates were 66.5% and 63.6%, and the 3-year and 5-year disease-free survival rates were 57.1% and 46.2%, respectively, in the 169 patients with malignant tumors. Conclusion: A new classification of maxillary defects is proposed, on which suitable flaps are selected to offer patients good functional and aesthetic outcomes and high quality of life.

Clinical comparative study of free posterior tibial artery perforator flap and radial forearm free flap for head and neck reconstruction / 中华耳鼻咽喉头颈外科杂志

Objective: To compare the clinical application results of the FPTF (free posterior tibial artery perforator flap) and RFFF (radial forearm free flap) for reconstruction of head and neck defects. Methods: A retrospective analysis of 27 cases treated with FPTF (19 males and 8 females, aged 14-69 years) and 24 cases with RFFF (11 males and 13 females, aged 22-69 years) for head and neck defect reconstruction at Beijing Tongren Hospital of Capital Medical University from January 2015 to December 2020 was conducted. Flap size, vascular pedicle length, matching degree of recipient area blood vessels, preparation time, total operation time, hospital stay, recipient area complications, donor area complications and scale-based patient satisfaction were compared between two groups of patients with FTPF and RFFF. SPSS 26.0 statistical software was used for statistical analysis. Results: There was no statistically significant difference between the two groups of patients in tumor T staging (P=0.38), primary sites (P=0.05) and mean flap areas ((53.67±29.84) cm2 vs. (41.13±11.08) cm2, t=-1.472, P=0.14). However the mean vascular pedicle length of FPTF was more than that of RFFF ((11.15±2.48)cm vs. (8.50±1.69)cm, t=-4.071, P<0.01). The donor sites of 4 patients in FPTF group could be sutured directly, while all the 24 patients in RFFF group received skin grafts from the donor sites. There was no statistically significant difference in the recipient area arteries between two groups of flaps (P=0.10), with more commonly using of the facial artery (RFFF: FPTF=21∶27), but there was significant difference in the recipient area veins (P<0.01), with more commonly using of the external jugular vein in RFFF (14/24) than FPTF (4/32) and the posterior facial vein in FPTF (27/32) than RFFF (9/24). There were 10 recipient complications and 3 donor complications in RFFF group; no recipient complication and 3 donor complications occurred in FPTF group. With patient's subjective evaluation of the donor site at 12 months after surgery, FPTF was better than RFFF (χ²=22.241, P<0.01). Conclusions: FPTF is an alternative to RFFF in head and neck reconstruction and has unique advantages in aesthetics and clinical application.

Retrospective study of thyroid invasion and central lymph node metastasis in 124 patients with hypopharyngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the characteristics of thyroid invasion and central lymph node metastasis of hypopharyngeal carcinoma, and the impact on survival rate and quality of life. Methods: A retrospective analysis of 124 cases (122 males and 2 females with age range from 36 to 78 years old) with laryngopharyngeal squamous cell carcinoma who were initially treated in the Department of Head and Neck Surgery, Beijing Tongren Hospital Affiliated to Capital Medical University from January 2014 to December 2017 was performed. The clinical data included tumor location, pathological T stage, pathological N stage, invasion of thyroid gland, central lymph node metastasis, surgical procedures and so on. Patients were grouped according to if presence of thyroid invasion and central lymph node metastasis. With follow-up, the survival was analyzed by Kaplan-Meier method, and tumor recurrence and metastasis were evaluated. Results: Of the patients, 12 patients had thyroid involvement and 5 patients had central lymph node metastasis. The incidence of thyroid involvement was 8.16% (8/98) in pyriform sinus, 1/18 in posterior pharyngeal wall and 3/8 in posterior cricoid wall, with statistically significant difference (χ2=15.076,P=0.008). The incidence of central lymph node metastasis was 1.02% (1/98) in pyriform sinus, 3/18 in posterior pharyngeal wall and 1/8 in posterior cricoid wall, also with statistically significant difference (χ2=11.205, P=0.008). There was no statistical correlation between thyroid invasion or central lymph node metastasis and gender, smoking or alcohol exposure history and tumor pathological differentiation (all P>0.05). The 3-year overall survival rate was 80.65% and the 3-year recurrence free rate was 85.48%. Totally 24 patients died in 3 years, including 4 cases in thyroid invasion group and 1 case in central lymph node metastasis group. Local recurrence occurred in 18 patients, including 4 cases in thyroid invasion group and 1 case in central lymph node metastasis group. There was no significant difference in survival between patients with and without thyroid invasion and central lymph node metastasis (all P>0.05). There were significantly difference in 3-year overall survival and relapse-free survival among the groups with different T stages, N stages, pathological stages and tumor pathological differentiation levels (all P<0.05). There were significantly differences in the levels of serum calcium and FT3 between the groups with or without thyroid invasion and central lymph node metastasis (all P<0.05). Conclusion: The incidences of thyroid invasion and central lymph node metastasis of hypopharyngeal carcinoma are rare, and the risk of occurrence is related to the primary site of tumor. Comprehensive evaluation, correct decision-making and accurate treatment could be helpful to cure radically the tumor, to prevent recurrence and to improve the quality of life of patients.

Evaluation of shoulder function after repairing head and neck defects with supraclavicular flap / 中华耳鼻咽喉头颈外科杂志

Objective: To evaluate the shoulder function in patients after repair of head and neck defects with supraclavicular flap. Methods: A retrospective analysis was performed on 56 patients (54 males, 2 females, aged 35-74 years old) who received the repair of head and neck defects with supraclavicular flaps at Department of Otorhinolaryngology Head and Neck Surgery of Beijing Tongren Hospital, Capital Medical University in January 2013-December 2020. The areas and types of flaps, disruption or infections of the incision at the donor sites and other postoperative complications were recorded. Quick disabilities of the arm, shoulder and hand (Quick-DASH) was used for evaluating the shoulder functions in 43 patients conforming to the standard for evaluation of the clinical functions of shoulders and upper limbs, to compare the postoperative upper limb functions between patients treated with clavicular flaps and patients with acromion flaps. Meanwhile, 30 patients who received bilateral neck lymph node dissection over the same period of time were selected for a comparative evaluation of the donor sides (observation group) and the opposite sides (control group). Data were processed with SPSS 22.0. Results: The areas of obtained supraclavicular flaps were (4-10) cm × (10-18) cm. Three patients (5%) showed the defect widths of 8-10 cm at the donor sites, which couldn't be sutured directly, received the repair of their shoulder defects with partial flaps. Defects in other patients were sutured directly. After surgery, 3 patients (5%) suffered from disruption of the acromion incision, which healed after 2 weeks of local dressing. The follow-up time was 6-43 (27±14) months. All patients expressed no dissatisfaction with the appearance. Among 43 patients, 28 (65%) were clavicular type and 15 (35%) were acromion type. The acromion type showed average motor ability and Quick-DASH scores higher than the clavicular type [(average motor ability: (14.4±4.7) vs. (11.8±3.1), t=2.105, P=0.048; Quick-DASH: (16.9±11.6) vs. (12.2±7.1), t=2.284, P=0.033]. Among 30 patients who received bilateral neck lymph node dissection over the same period of time, the observation group showed higher average motor ability, local symptoms and Quick-DASH scores than the control group [average motor ability: (13.4±5.8) vs. (9.8±4.2), t=3.024, P=0.004; average local symptoms: (4.1±1.0) vs. (3.4±1.0), t=2.537, P=0.014; Quick-DASH: (15.6±14.7) vs. (5.2±11.1), t=3.106, P=0.003]. Conclusion: Shoulder dysfunction exists after treatment with supraclavicular flap, which is related to the flap type.

miR-18a enhances the radiosensitivity of nasopharyngeal carcinoma cells through inducing autophagy / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the impacts of miR-18a overexpression or depression on the radiosensitivities of nasopharyngeal carcinoma cell line CNE1 and CNE2 and underlying mechanisms. Methods: CNE1 and CNE2 were transfected with miR-18a mimics, inhibitor and the corresponding control vectors. qRT-PCR and western blot were used to determine the ataxia telangiectasia mutated (ATM) expressions in CNE1 and CNE2. CNE1 and CNE2 with stably expressing miR-18a and miR-18a siRNA were constructed. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the impacts of the miR-18a overexpression or depression combined with irradiation on the cell growth. Flow cytometry was used to detect the cell apoptosis and cell cycle. Colony formation assay was used to evaluate the raodiosensitivities of cells. Acridine orange (AO) staining and western blot were used respectively to test the autophagy and the expressions of related proteins. Independent samples t test was used to compare the mean value between groups by using SPSS 16.0. Results: ATM mRNA was decreased significantly in CNE1 and CNE2 cells transfected with 100 or 200 nmol/L miR-18a mimics for 48 hours (CNE1: RQ=0.174±0.139 and 0.003±0.001, t=9.939 and 19 470.783;CNE2: RQ=0.024±0.008 and 0.019±0.012, t=270.230 and 137.746, respectively, all P<0.001). ATM proteins were also decreased after transfected with 100 or 200 nmol/L miR-18a mimics for 72 hours. While in the cells transfected with 100 and 200 nmol/L miR-18a inhibitor for 48 hours, the expressions of ATM mRNA were upregulated significantly (CNE1: RQ=9.419±2.495 and 2.500±1.063, t=-4.427 and -41.241; CNE2: RQ=7.210±0.171 and 115.875±15.805, t=-62.789 and -12.589, all P<0.05), and the expressions of ATM proteins increased after transfected for 72 hours. The growth of cells with miR-18a overexpression plus 4 Gy irradiation were obviously inhibited compared to that of cells with the 4Gy irradiation alone; while the growth of miR-18a-inhibited cells increased compared to that of cells with 4 Gy irradiation alone (all P<0.05). CNE1 transfected with 100 nmol/L miR-18a mimics plus 4 Gy irradiation showed the higher apoptosis rate than the cells with 4 Gy irradiation alone ((22.9±2.1)% vs. (16.3±1.0)%, t=-4.870, P<0.01). Compared to the cells with 4 Gy irradiation alone, miR-18a-overexpressed cells plus 4 Gy irradiation decreased their percentages in G1 phases ((20.2±3.0)% vs. (29.8±4.4)%, t=3.119) and G2/M phases ((21.5±0.9)% vs. (33.4±3.1)%, t=6.410, P<0.05 for both), and increased their percentages in S phases ((56.7±4.9)% vs. (36.8±6.4)%, t=-4.246, P<0.05), and these cells possessed less colony number after exposure to different doses of irradiation, more autophagy-lysosome number, and more expressions of LC3 proteins (all P<0.05). There were no significant differences in the expressions of p62 expressions between different groups of cells. Conclusion: Overexpression of miR-18a can enhance the radiosensitivities of NPC cells by targeting ATM to abrogate G1/S, G2/M arrest and to induce autophagy and apoptosis.

The expression of Notch pathway in nasal polyps and its correlation with the decreased Treg percentage and eosinophil infiltration / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the correlation between Notch pathway expression in nasal polyps and Treg percentage and Eos infiltration. Methods: Patients with chronic sinusitis and simple nasal septum deviation who received nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-Sen University between November 2012 and August 2018 were selected and enrolled in CRS group and control group respectively. Nasal mucosa tissues were collected from 30 CRSsNP patients (14 males and 16 females aged from 18 to 63), 58 CRSwNP patients (38 males and 20 females aged from 18 to 65) and 29 patients (19 males and 10 females aged from 20 to 57), who underwent nasal endoscopic surgery for correction of simple nasal septum deviation. Hematoxylin-eosin(HE) staining was used to observe the infiltration of eosinophilic granulocytes in the tissues and to classify chronic sinusitis with polyps (CRSwNP) into eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP)and non-eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Notch pathway receptors (Notch-l, 2, 3, 4) and their ligands (Jagded-l, Jagded-2, Delta-l, Delta-3and Delta-4) in the nasal mucosa of each group, as well as the expression of Th2 cytokines (IL-4, IL-5, IL-13), eosinophilic cationic protein (ECP)and the key transcription factor Foxp3 in Treg cells. Finally, flow cytometry was used to detect CD4+CD25+Foxp3+ Treg cells in nasal mucosa of each group. Results: Compared with controls, the expression of Th2 cytokines (IL-4, IL-5, IL-13) in CRSsNP and non-Eos-CRSwNP patients was the highest in Eos-CRSwNP (F=16.930,9.197,9.116, all P<0.05). Foxp3 had the lowest expression in Eos-CRSwNP patients and was lower than non-Eos-CRSwNP patients (F=2.780,P<0.05), and was negatively correlated with ECP (r=-0.326,P<0.05). Compared with controls, Eos-CRSwNP patients in CRSsNP patients and non-Eos-CRSwNP patients exhibited a significantly lower frequency of CD4+CD25+Foxp3+Treg cells (F=13.140, all P<0.01). The expression of Notch-l and Jagged-l in Eos-CRSwNP was significantly higher than that of the controls, CRSsNP patients and non-Eos-CRSwNP patients (F=5.953/F=6.380, P<0.05). In the nasal polyp group, the expression of Notch-l and Jagged-l showed significantly negative correlation with Foxp3 (r=-0.611/-0.346, all P<0.05), and positive correlation with Th2 cytokines (IL-4, IL-5, IL-13) and ECP, respectively (r=0.781/0.459,0.621/0.601,0.605/0.490,0.464/0.668, all P<0.05). There was no significant difference in the expression of receptor and ligand of the other Notch pathway among the groups. Conclusion: Abnormal activation of Notch-l/Jagged-l pathway may be involved in decreasing Treg ratio in Eos-CRSwNP, thereby promoting Th2 inflammatory response and Eosinophil infiltration.

Therapeutic effect of acupuncture combined with medication on chronic prostatitis of damp and heat stasis / 中国针灸

OBJECTIVE@#To observe the clinical therapeutic effect of electroacupuncture (EA) combined with tamsulosin hydrochloride sustained release capsule on chronic prostatitis (CP) of damp and heat stasis.@*METHODS@#A total of 70 patients with CP of damp and heat stasis were randomized into an acupuncture plus medication group (35 cases, 4 cases dropped off) and a medication group (35 cases, 5 cases dropped off). In the medication group, tamsulosin hydrochloride sustained release capsule was given orally, 0.2 mg a time, once each night. On the basis of treatment in the medication group, EA was applied at Guanyuan (CV 4), Sanyinjiao (SP 6) and Yinglingquan (SP 9), with disperse-dense wave, 5 mA in intensity for 30 min. Treatment for 30 days was as one course, and totally 3 courses were required in both groups. Before treatment, 1, 2, 3 months into treatment and at the follow-up of 2 months after treatment, the TCM syndrome score and National Institutes of Health chronic prostatitis symptom index (NIH-CPSI) score were observed, and the clinical efficacy was evaluated in both groups.@*RESULTS@#Compared before treatment, the TCM syndrome scores of 3 months into treatment and follow-up were decreased in the acupuncture plus medication group (@*CONCLUSION@#Acupuncture combined with medication can improve the clinical symptoms in patients with CP of damp and heat stasis, and its therapeutic effect is superior to simple western medication.

Relationship of adrenal insufficiency with metabolic crisis and sudden death / 中国实用儿科杂志

Adrenal insufficiency(AI)is a type of adrenocortical hormone-deficient lesion caused by primary or secondary factors. Because of its lack of specificity in clinical manifestations,it is difficult to identify early,which may lead to delayed diagnosis. If the treatment is not timely,it may cause adrenal crisis(AC)or even sudden death. Sudden death in children due to adrenal insufficiency may be directly related to hyperkalemia caused by adrenal crisis. Gastrointestinal illness and infections are common precipitant for an adrenal crisis. Despite it being a treatable condition for now,failure of adequate preventive measures or delayed treatment has often led to unnecessary deaths. Therefore,more measures are needed to prevent the adrenal crisis and to ensure that appropriate emergency medical services are established after the crisis to reduce the rate of sudden death.

Construction of Human Nasophayngeal Cancer Cells Stablely Expressing miR-18a and miR-18a siRNA / 中山大学学报(医学科学版)

[Objective]To construct miR-18a overexpression and inhibition lentivirus vectors and to determine their effects on human nasopharyngeal cancer(NPC)cell line CNE1 and CNE2.[Methods]Designed the primers for Real-time polymerase chain(PCR)reaction to obtain the miR-18a premature gene.The premature gene and the siRNA oligo-nucleutides of miR-18a were connected to the lentivirus vector GV369 and GV280,respectively.The construction vectors were confirmed by DNA sequencing.Then,293T cell was infected with the vectors plus Helper 1.0 and pHelper 2.0 vec-tors to obtain recombinant lentivirus vector for miR-18a overexpression and inhibition. The NPC cell line CNE1 and CNE2 were infected with the successful recombinant lentivirus vectors.Puromycin was added to select the positive infect-ed cells. PCR method was used to detect the miR-18a expression level after infecting the recombinant lentivirus vector into the NPC cell line.[Results]A recombinant lentivirus vector expressing miR-18a interference oligonucleutides was obtained and confirmed by DNA sequencing.The virus titer was 3×108TU/mL,and the expression of its target gene ATM was downregulated in CNE1 and CNE2.A recombinant lentivirus vector expressing miR-18a premature gene was obtained and confirmed by DNA sequencing. The virus titer was 3×108TU/mL,and the miR-18a was overexpressed in CNE1 (20.3 fold upregulation,P<0.01)and CNE2(122.5 fold upregulation,P<0.01),and its target gene ATM was downregu-lated.[Conclusions]The miR-18a overexpression and suppression lentivirus vectors are successfully constructed.These vec-tors could alter the expression level of miR-18a in NPC cell line significantly,and provide a stable cell line for functional studies in the future.

Comparison of different bowel preparation methods for capsule endoscopy / 中国内镜杂志

Objective To explore the optimal bowel preparation for capsule endoscopy (CE). Methods 102 patients were recruited for CE and randomly divided into 3 groups. The group A (n = 40) : patients received polyethylene glycol electrolyte powder (PEG) 137.12 g dissolved in 2 000 ml water at 21:00 one day prior to CE, and taken PEG 68.56 g dissolved in 1 000 ml of water four hours before the procedure. Group B (n = 32): patients received PEG 205.68 g dissolved in 3 000 ml of water four hours prior to CE. Group C (n = 30): patients used a 500 ml 20% mannitol and 2 500 ml water bowel preparation four hours prior to CE. All patients were treated with 120 mg simethicone immediately after swallowing CE. The incidences of adverse events, small-bowel preparation quality and transit time were analyzed. Results The adverse effects rate in each group was similar (15.00%, 15.63% vs 16.67%, P > 0.05). The small-bowel preparation quality was better in both B and C groups than A group (P < 0.05). In the C Group, small-bowel preparation quality was slightly better than the B group, but the difference was not statistically significant (P > 0.05). In comparison with patients in both B and C groups, those in A group had a longer small-bowel transit time (P < 0.05), whereas there was no significant difference between B and C group (P > 0.05). Conclusion Bowel cleansing effect was better in single dose regimen than split dose protocol. The single dose regimen of 500 ml 20% Mannitol for bowel preparation is suitable prior to CE.

Spectral domain optical coherence tomography quantifies the macular thickness in patients with type 2 dia betes mellitus without significant diabetic retinopathy / 眼科新进展

Objective To observe the retinal thickness of macular in type 2 diabetic mellitus (T2DM) patients without clinical features of diabetic retinopathy (DR).Methods Totally 40 patients (40 eyes) with T2DM without DR and 70 healthy volunteers (70 eyes) from August 2017 to October 2017 were enrolled in this study.Usage of spectral domain optical coherence tomography (SD-OCT) and the software of automatic segmentation to measure the average thickness of total retinal (R),inner retinal layer (IRL) and outer retinal layer namely photoreceptor layer (PL) in macular.The foveal center was divided according to three concentric circle with the diameter of 1 mm,3 mm and 6 mm (including the partition of R total,R-1,R-3,R-6,IRL-1,IRL-3,IRL-6,PL-1,PL-3 and PL-6),and the average retinal thicknesses of these partitions between these two group were compared and analyzed.Results The thickness of PL-1 and PL-3 in no DR group was significantly thinner than that in the normal control group [(71 ± 4)μm vs.(73 ± 3) μm and (66 ± 2) μm vs.(67 ± 2) μm,respectively] (both P ＜ 0.05).In the normal control group,except the IRL-6 and PL-1,the difference of the thickness was significant in the other macular regions between various genders (all P ＜ 0.05).In the male subjects,the thickness of PL-3 and PL-6 in no DR group was significantly thinner than that in the normal control group [(67 ± 2) μm vs.(68 ± 2) μm and (65 ± 2) μm vs.(66 ± 2) μm,respectively] (both P ＜ 0.05).In the female subjects,the mean thicknesses of PL-3 and PL-6 in no DR group was significant thinner than those in normal control group [(65 ± 2) μm vs.(67 ± 2) μm and (63 ± 2) μm vs.(64 ± 2) μm,respectively] (both P ＜ 0.05).There was no obviously difference in the other parts between these two groups,Conclusion The mean retinal thicknesses of the parafovea and perifovea are significantly thinner in no DR group than that of the normal control group.The measurement of the PL thickness of macular by SD-OCT may promote the study of early stage of DR,and become an important biological marker for early monitoring of DR.

Relationship of Serum Neutrophil Gelatinase-associated Lipocalin Levels with Stroke Attack and Renal Dysfunction in Patients / 现代检验医学杂志

Objective To explore the correlations of neutrophil gelatinase associated lipocalin (NGAL) with cerebral stroke attack and renal dysfunction of patients.Methods 63 patients with first-diagnosed hemorrhagic stroke (HS),233 patients with first diagnosed ischemic stroke (IS) and 293 healthy controls were randomly selected.Their age,gender and clinical his tory of hypertension,diabetes,coronary heart disease and hyperuricemia were recorded at admission,and their serum levels of kidney function/injury biomarkers (NGAL,Cr,Urea,CysC and eGFR) were measured at the next day.According to KDIGO guidelines standards,all subjects were divided into four groups on behalf of their renal damage degree:AKI,CKD,transient or no kidney injury,and control group.And then,the relations of these indicators with stroke attack and renal function of HS and IS patients were analyzed.Results The medians of serum NGAL levels in the patients with HS and IS were 160.0 and 142.9 μg/L,their difference had no statistical significance (Z=2.332,P＞0.05),but were higher than healthy controls (Z =12.621,9.189,all P＜0.05).The levels of serum NGAL in AKI,CKD,transient or no renal dysfunction,and control groups were 375.3,228.6,141.6 and 103.8 μg/L respectively,which had statistically significant differences between each other (Z=3.661～11.237,all P＜0.05).NGAL level was positively related to HS attack (rp=0.423,P＜0.001),IS attack (rp =0.231,P＜0.001) and renal dysfunction in both HS and IS patients (rp=0.429,0.289,all P＜0.001).Its partial cor relation coefficient was only lower than hypertension within HS stroke attack,and lower than coronary heart disease and diabetes within IS stroke attack,but the highest within renal function of either HS or IS patients.For assessing HS and IS attack,NGAL levels play a limited role (OR=1.044,0.974,allP＜0.001),in contrary to NGAL abnormity (OR =27.841,15.411,all P＜0.001).Conclusion Elevated NGAL levels may be a risk factor for stroke,suggesting that kidney injury or renal dysfunction may be associated with stroke,and the prevention and treatment of stroke should pay attention to the change of NGAL.

Reye syndrome and sudden death symptoms after oral administration of nimesulide due to upper respiratory tract infection in a boy / 中国当代儿科杂志

A boy aged 6 years and 3 months developed upper respiratory tract infection and pyrexia 2 months ago and was given oral administration of nimesulide by his parents according to directions. Half an hour later, the boy experienced convulsions and cardiopulmonary arrest, and emergency examination found hypoketotic hypoglycemia, metabolic acidosis, significant increases in serum aminotransferases and creatine kinase, and renal damage. Recovery of consciousness and vital signs was achieved after cardiopulmonary resuscitation, but severe mental and movement regression was observed. The boy had a significant reduction in free carnitine in blood and significant increases in medium- and long-chain fatty acyl carnitine, urinary glutaric acid, 3-hydroxy glutaric acid, isovalerylglycine, and ethylmalonic acid, suggesting the possibility of multiple acyl-CoA dehydrogenase deficiency. After the treatment with vitamin B2, L-carnitine, and bezafibrate, the boy gradually improved, and reexamination after 3 months showed normal biochemical parameters. The boy had compound heterozygous mutations in the ETFDH gene, i.e., a known mutation, c.341G>A (p.R114H), from his mother and a novel mutation, c.1484C>G (p.P495R), from his father. Finally, he was diagnosed with multiple acyl-CoA dehydrogenase deficiency. Reye syndrome and sudden death symptoms were caused by nimesulide-induced acute metabolic crisis. It is concluded that inherited metabolic diseases may be main causes of Reye syndrome and sudden death, and biochemical and genetic analyses are the key to identifying underlying diseases.

Gene chip technology for detecting mycobacteria and Mycobacterium tuberculosis resistance of isoniazid and rifampin in the application of tuberculous empyema / 中国人兽共患病学报

To evaluate the clinical value of gene chip technology (GCT) in detecting the mycoba-cteria,isoniazid and rifampin resistance of patients diagnosed tuberculous empyema.The 182 patients who met the inclusion criteria were enrolled to this study from January 2011 to December 2015,whose pus mycobacterial species were detected by GCT and MGIT,the simultaneous and sensitivity of them were compared.Meanwhile,36 patients diagnosed tuberculous empyema were selected to detect isoniazid and rifampin resistance.The simultaneous and sensitivity of GCT were evaluated base on the standard of MGIT.The 135 patients were diagnosed by tuberculous empyema.The specificity of GCT was same to MGIT (95.7％),the the sensitivity was 48.9％ (66/135)in GCT,26.7％ in MGIT,there was significant difference between them (x2=80.5,P＜ 0.05).The sensitivity,specificity and coincidence rate of GCT in rifampin resistance were 100％,the sensitivity,specificityand coincidence rate in INH were 50.0％(1/2),97.1％(33/34) and 94.4％.Gene chip technology for detection of mycobacteria has high sensitivity and specificity,which can identify non-tuberculous mycobacteria quickly.And it can also effectively detect the resistance of Mycobacterium tuberculosis to isoniazid and rifampicin.It has important significance in early diagnosis and treatment of tuberculous empyema.

High-frequency echocardiography for assessment of regional wall motion abnormality and cardiac function in mice with myocardial infarction / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the value of high-frequency echocardiography in assessing cardiac structure and function in a mouse model of myocardial infarction.</p><p><b>METHODS</b>Twenty-five C57BL/6 mice were randomly divided into sham-operated group (n=10) and myocardial infarction model group (n=15) established by ligation of the left anterior descending artery. The cardiac structure, regional wall motion and cardiac function of mice were examined with pulsed wave Doppler (PWD), tissue Doppler imaging (TDI), EKV and M-mode echocardiography 3 days before and at 1 week after the operation. The histological changes and myocardial structure of the heart were observed at 1 week after the operation.</p><p><b>RESULTS</b>High-frequency echocardiography and HE staining detected obvious myocardial infarction in the mice in the model group. Compared with the sham-operated mice, the mice with myocardial infarction showed significant left ventricular expansion, obvious thinning of the ventricular wall, and significantly decreased ventricular systolic function and diastolic function with regional wall motion abnormality and ventricular remodeling.</p><p><b>CONCLUSION</b>s 2D-type echocardiography combined with M-mode, PWD, TDI and EKVTM for allows accurate and sensitive detection of the loci and severity of myocardial infarction to provide important evidence for clinical diagnosis and treatment of myocardial infarction.</p>

Effect of atopy on serum glucocorticoid receptor levels in children with bronchiolitis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of atopy on the expression of glucocorticoid receptors in children with bronchiolitis.</p><p><b>METHODS</b>ELISA was used to measure the changes in the serum levels of glucocorticoid receptor α (GRα) and glucocorticoid receptor β (GRβ) in the bronchiolitis group (77 children, including 34 children with atopy) and pneumonia group (68 children). Thirty-eight children who were prepared to undergo surgeries for non-infectious diseases and had no atopy or family history of allergic diseases were enrolled as the control group.</p><p><b>RESULTS</b>The bronchiolitis group and the pneumonia group had significant increases in the serum levels of GRα and GRβ compared with the control group (P<0.01), and the bronchiolitis group had significant increases in these levels compared with the pneumonia group (P<0.01). Compared with the control group and the pneumonia group, the bronchiolitis group had a significant increase in the GRα/GRβ ratio (P<0.01). Compared with the control group, the children with or without atopy in the bronchiolitis group had significant increases in the serum levels of GRα and GRβ (P<0.01). The non-atopic children in the bronchiolitis group had a significant increase in the serum level of GRβ compared with the atopic children (P<0.01). The atopic children in the bronchiolitis group had a significant increase in the GRα/GRβ ratio compared with the control group and non-atopic children in the bronchiolitis group (P<0.01).</p><p><b>CONCLUSIONS</b>Children with bronchiolitis have increased serum levels of GRα and GRβ. The children with atopy have an increased GRα/GRβ ratio, suggesting that the atopic children with bronchiolitis are highly sensitive to glucocorticoids.</p>