TLR4-MyD88-NF-κB signaling imbalances Th17 and Treg cells in thymoma with myasthenia gravis.

OBJECTIVE: Thymus is an immune organ in which pathological changes may cause autoimmune diseases, including myasthenia gravis (MG). Recent studies have focused on Toll-like receptor 4 (TLR4) signaling as the cause of such changes. In our previous study, an imbalance of T helper 17 (Th17) cells and T regulatory (Treg) cells was found in MG thymoma. These results suggest the involvement of TLR4 in the pathogenesis of thymoma MG via an alteration of the Th17/Treg balance. Here, we aimed to assess whether the TLR4-MyD88-NF-κB pathway is upregulated in MG thymoma and its relationship with Th17/Treg cells. PATIENTS AND METHODS: We collect thymoma samples from 54 patients with or without MG, detecting the expression level of TLR4, MyD88, and NF-κB in thymoma tissues. Next, we established an in vitro experiment of coculturing thymoma cells with CD4+ T cells and detected the differentiation of Th17 cells and Treg cells and their marker protein, retinoid-related orphan receptor gamma t (RORγt) and forkhead transcription factor 3 (Foxp3). RESULTS: We found TLR4, MyD88, and NF-κB expressed more in MG thymoma compared with simple thymoma. After the transwell coculturing, we observed an imbalance of Th17/Treg cells after TLR4 stimulation. CONCLUSIONS: TLR4 is stimulated in thymoma, causing an increase of Th17 cells and a decrease of Treg cells, namely an imbalance of Th17/Treg cells, resulting in MG.

[Clinical management and prognosis for descending necrotizing mediastinitis].

Objective: To investigate the clinical characteristics, treatment experiences and prognostic factors for descending necrotizing mediastinitis (DNM). Methods: A retrospective analysis was performed on the data of 22 patients with DNM diagnosed and treated in Henan Provincial People's Hospital from January 2016 to August 2022, including 16 males and 6 females, aged 29-79 years. After admission, all patients underwent CT scanning of the maxillofacial, cervical, and thoracic regions to confirm their diagnoses. Emergency incision and drainage were performed. The neck incision was treated with continuous vacuum sealing drainage. According to the prognoses, the patients were divided into cure group and death group, and the prognostic factors were analyzed. SPSS 25.0 software was used to analyze the clinical data. Rusults: The main complaints were dysphagia (45.5%, 10/22) and dyspnea (50.0%, 11/22). Odontogenic infection accounted for 45.5% (10/22) and oropharyngeal infection accounted for 54.5% (12/22). There were 16 cases in the cured group and 6 cases in the death group, with a total mortality rate of 27.3%. The mortality rates of DNM typeⅠand typeⅡwere respectively 16.7% and 40%. Compared with the cured group, the death group had higher incidences for diabetes, coronary heart disease and septic shock (all P<0.05). There were statistically significant differences between the cure group and the death group in procalcitonin level (50.43 (137.64) ng/ml vs 2.92 (6.33) ng/ml, M(IQR), Z=3.023, P<0.05) and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score (16.10±2.40 vs 6.75±3.19, t=6.524, P<0.05). Conclution: DNM is rare, with high mortality, high incidence of septic shock, and the increased procalcitonin level and APACHE Ⅱ score combined diabetes and coronary heart disease are the poor prognostic factors for DNM. Early incision and drainage combined with continuous vacuum sealing drainage technique is a better way to treat DNM.

Relationships between diabetic nephropathy and insulin resistance, inflammation, Trx, Txnip, CysC and serum complement levels.

OBJECTIVE: To investigate the relationships between diabetic nephropathy (DN) and insulin resistance, inflammation, thioredoxin (Trx), thioredoxin-interacting protein (Txnip), Cystatin C (CysC) and serum complement levels. PATIENTS AND METHODS: A total of 119 patients with type 2 diabetes mellitus (T2DM) treated in the Endocrinology Department of our hospital from January 2017 to December 2017 were enrolled as the experiment group, while 30 healthy volunteers were selected as the control group. The expression levels of inflammatory factors, Trx, Txnip, CysC and serum complements in every subject were detected. In addition, the type 2 diabetic nephropathy rat model was established via high-fat diet and injection of low-dose streptozotocin. Blood glucose, insulin resistance indexes and 24h-urinary albumin excretion were measured, and the histomorphological characteristics of the kidney in animals were observed. RESULTS: In clinical subjects, Trx level was notably lower in the simple DM group, early DN group and clinical DN group in comparison with that in the control group. The levels of Txnip and CysC in the simple DM group, early DN group and clinical DN group were remarkably higher than those in the control group. Moreover, the expression levels of TNF-α and IL-6 in the clinical DN group were significantly elevated compared with those in the simple DM group and early DN group. In addition, C1q expression in the clinical DN group was higher than that in the simple DM group and early DN group. In model rats, HOMA-IR was distinctly higher in the DM group and DN group than that in the control group. The ratio of kidney weight to body weight (KW/BW) was evidently higher in the DN group in comparison with that in the control group and DM group. CONCLUSIONS: Insulin resistance, inflammatory factors, and levels of Trx, Txnip, CysC and serum complement C1q are related to the progression of DM.

Clinical significance of tumor mutation burden and DNA damage repair in advanced stage non-small cell lung cancer patients.

OBJECTIVE: This study aimed to investigate the impact of tumor mutational burden (TMB) and DNA damage repair (DDR) gene alteration on overall survival (OS) in advanced non-small cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: A DNA library of cancer cells from 67 NSCLC patients in stages III-IV was constructed for next-generation sequencing (NGS). Geneseeq422 probes were used for hybridization enrichment. The target-enriched library was sequenced on HiSeqNGS platforms, and we analyzed the relevant signaling pathways. Then, we correlated the OS of the patients with TMB and DDR mutations. RESULTS: Many significant alterations were found, including in the EGFR, p53, KRAS, RB1, ERBB2, NF1, DNMT3A, ALK, MYC, PIK3CA, ROS1, BRAF, ARID1A, PTEN, CDKN2A, and FGF19 genes. We also identified many mutations in the genes relevant to the DDR pathway. Interestingly, we found that the TMB of patients with DDR gene mutations was dramatically higher than that in the DDR wild-type (WT). Univariable analysis showed that DNMT3A, RB1, DDR pathway-related gene mutations, and TMB were critical factors for the effects on OS. Multivariable analysis confirmed that DNMT3A and mutations in the DDR pathway-related genes were important for predicting OS. CONCLUSIONS: Multiple mutations in the genes of the DDR pathway caused higher TMB levels, which resulted in longer OS. By contrast, OS was significantly longer in patients with non-DNMT3A mutations than in those with DNMT3A variants. DNMT3A alteration in NSCLC patients led to poor outcomes.

[Clinical analysis of mediastinal lymph node dissection through sternotomy approach in the treatment of papillary thyroid carcinoma with mediastinal lymph node metastasis].

Objective: To investigate the rule of mediastinal lymph node metastasis of papillary thyroid carcinoma and the application of therapeutic mediastinal lymph node dissection through the sternotomy approach in the treatment of mediastinal lymph node metastasis of papillary thyroid carcinoma. Methods: All cases of papillary thyroid carcinoma with mediastinal lymph node metastasis treated through sternotomy cooperated by thoracic surgeons and head and neck surgeons from January 2006 to January 2017 in Cancer Hospital of Chinese Academy of Medical Sciences were included in this study. The distribution, metastasis rate, metastasis degree, surgical method, surgical complications and postoperative survival of patients with mediastinal lymph node metastasis were retrospectively analyzed. Results: A total of 31 patients (16 males and 15 females) with papillary thyroid cancer with mediastinal lymph node metastasis, with a median age of 46 (19-65) years, were enrolled in the group. Partial upper sternotomy was used in 28 cases, and total sternotomy was used in 3 cases. The mediastinal lymph nodes of papillary thyroid carcinoma metastasized farthest to the station 6, and the lymph node metastasis rate of each group from high to low was: 2R (61%), 1R (39%), 3A (39%), 1L (16%), 2L (10%), 4R (10%), 5 (3%) and 6 (3%). No metastasis was observed in station 3P, 4L and 7. In addition, the degree of lymph node metastasis at station 2R was the highest, reaching 35% (77/219). Extra-nodal invasion of mediastinal metastatic lymph nodes in thyroid papillary carcinoma is common (23%), easily fuses into masses (23%) and invades peripheral vascular nerves (26%). Up to 29% of blood transfusions are required during or after surgery due to oozing or bleeding (9/31). The 1-, 3-, 5-and 10-year survival rates of patients undergoing surgical treatment were 94%, 94%, 87% and 81%, respectively. Conclusion: Papillary thyroid carcinoma can metastasize to almost all mediastinal lymph nodes except station 3P, 4L and 7. Radical mediastinal lymph node dissection through sternotomy is an effective method for the treatment of mediastinal lymph node metastasis of thyroid papillary carcinoma.

[The prevalence and relationship between laryngopharyngeal reflux disease, anxiety and depression in otolaryngology outpatients].

Objective: To obtain the prevalence laryngopharyngeal reflux disease (LPRD), anxiety and depression in otorhinolaryngology outpatients and to explore the role of mental and psychological factors (anxiety and depression) in their pathogenesis. Methods: A questionnaire survey of reflux symptom index(RSI) scale and hospital anxiety and depression (HAD) scale were used to report 1 111 cases of outpatients in Department of Otorhinolaryngology Head and Neck Surgery, Peking University People's Hospital, from July 2017 to June 2018 (486 males, 625 females, age of 18-96 years old, median age of 38[30,53] years old) and to obtain the prevalence of LPRD, anxiety and depression. RSI-positive patients were selected in the case group, and RSI-negative patients were selected in the control group. The differences in HAD scores between the two groups were compared, and the risk factors of laryngopharyngeal reflux were analyzed. Statistical analysis was performed using SPSS 20.0 software. Results: There were 151 cases in the case group and 960 cases in the control group. The prevalence of LPRD was 13.59% (151/1 111).There was no significant difference in the prevalence of LPRD between different genders (P>0.05). The prevalence rate was the highest in the 18-40 age group, and the difference in the prevalence of all age groups (18~ 40 years old; 41-65 years old; >65 years old) was statistically significant (P<0.05). The prevalence of LPRD among smokers and non-drinkers was higher than that of non-smokers and non-drinkers and the prevalence of the two groups was statistically significant (P<0.05). The most common symptoms of the RSI scale were pharyngeal foreign body sensation (92.72%,140/151), persistent clearing throat (88.74%,124/151), excessive sputum or nasal reflux (82.12%, 124/151). There were significant statistical differences between the two groups (P<0.05). Ninty-one patients with anxiety, the prevalence was 8.19%(91/1 111); 76 patients with depression, the prevalence was 6.84%(76/1 111).Among the LPRD patients, the hospital anxiety scale scored 29.14% (44/151), and the hospital depression scale scored 17.22% (26/151). The scores of anxiety symptoms and depressive symptoms in the LPRD group were higher than those in the non-LPRD group. The above scores were statistically significant (P<0.05). Logistic regression analysis showed that smoking, anxiety and symptoms of gastroesophageal reflux disease were independent risk factors for laryngopharyngeal reflux. Conclusions: The prevalences of LPRD, anxiety and depression in the otorhinolaryngology clinic are 13.59%, 8.19% and 6.84%, respectively. Among patients with laryngopharyngeal reflux, the prevalence of anxiety is 29.14%, and the prevalence of depression is 17.22%. Age, smoking, drinking, alcohol consumption, education level, course of disease, symptoms of gastroesophageal reflux disease, pharyngeal foreign body sensation, etc. are related to LPRD. Mental factors (anxiety and depression) may play a role in LPRD. Smoking, anxiety symptoms and symptoms of gastroesophageal reflux disease are closely related to the incidence of LPRD.

[Investigation of laryngopharyngeal reflux prevalence and risk factors in otolaryngology].

Objective:To investigate the prevalence and related risk factors of laryngopharyngeal reflux in otolaryngology. Method:During January 2019 to March 2019, the inpatients in otolaryngology were investigated by the questionnaire of reflux symptom index scale and the laryngopharyngeal reflux related risk factors were analyzed. Result:Among the 227 patients, 33 patients with suspected LPR contained 19 patients（20.7%） of 92 patients in the laryngopharyngeal group, 10 patients（16.1%） of 62 patients in the nasal group, and 4 patients（5.5%） of 73 patients in the ear group. LPR prevalence in the laryngopharyngeal group was statistically different from that in the ear group（P<0.05）. Univariate logistic regression analysis showed that age, gender, smoking, drinking and BMI were risk factors of LPR（P<0.01）. Multivariate logistic regression analysis indicated that BMI was an independent risk factor of LPR（P<0.01）. Conclusion:LPR had a high prevalence rate in otolaryngology related diseases, and appropriate synergistic anti-LPR treatment could be carried out while treating otolaryngology related diseases.

[Establishment and research of a New Zealand rabbit model of laryngopharyngeal reflux].

Objective: To establish a New Zealand rabbit animal model of laryngopharyngeal reflux disease (LPRD) using esophageal balloon together with metal internal stent dilation and to investigate the changes of mucosa. Methods: 20 New Zealand rabbits were randomly divided into experimental group and control group, with 10 in each group. Balloon dilatation and metal internal stent dilation were carried out in experimental group to reproduce the animal model of LPRD.The middle of balloon was placed at the lower esophageal sphincter (LES) while the stent was placed at the upper esophageal sphincter (UES). The guide wire was placed in the control group, but the balloon was not expanded and the stent was not placed. The general condition, pH value of hypopharynx, laryngeal histopathology and changes of pepsin content of New Zealand rabbits were observed regularly. The difference between experimental group and control group was compared. Results: The 24-hour Dx-pH monitoring results showed that the number of reflux episodes(20.0[9.5, 35.0], 13.0[6.5, 22.0]), and the percent time below pH 5.5 (1.36%[0.60%, 4.57%], 1.36%[0.43%, 2.77%]) in the experimental group at the 2nd and 4th week were significantly different from those in the control group (0[0,3.0], 1.0[0.5, 3.8]; 0[0, 0.01%], 0[0, 0], respectively, all P<0.01), suggesting that the experimental group New Zealand rabbits developed LPRD. Compared with the control group under microscope, lymphocytes infiltration and submucosal gland hyperplasia increased in the mucosa of the throat of the experimental group. The results of pepsin immunohistochemical staining between the two groups were statistically significant (P=0.014). Conclusion: The use of balloon dilatation of the LES combined with metal stent dilatation of the UES can successfully establish a laryngopharyngeal reflux model, and lesions in the throat tissue can be observed.

[An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels].

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People's Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories' results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion: The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.

Reduced serum miR-98 predicts unfavorable clinical outcome of colorectal cancer.

OBJECTIVE: Circulating microRNAs (miRNAs) are considered to be promising biomarkers for the diagnosis and prognosis prediction of cancers. However, the potential clinical significance of the serum miR-98 in colorectal cancer (CRC) remained unclear. This study aimed to examine the serum miR-98 levels in CRC patients and explore its potential value for CRC. PATIENTS AND METHODS: A total of 115 CRC cases and 50 healthy volunteers were enrolled in this study. Quantitative reverse-transcription PCR (qRT-PCR) was performed to detect serum miR-98 expression in all the participants. RESULTS: The results revealed that serum miR-98 levels were frequently downregulated in CRC patients compared with controls. In addition, low serum miR-98 levels were closely associated with aggressive clinical features and shorter survival. Receiver operating characteristic (ROC) curve analysis demonstrated that serum miR-98 could well differentiate CRC patients from healthy controls with relatively high accuracy. Multivariate analysis further demonstrated that serum miR-98 was an independent prognostic factor for both overall survival and disease-free survival. Mechanistically, MYC, IL-6, and HIST1H2BH were identified as direct downstream targets of miR-98 in CRC cells. CONCLUSIONS: Collectively, serum miR-98 might be useful as an indicator for predicting the clinical outcome of CRC patients.

MiR-647 promotes proliferation and migration of ox-LDL-treated vascular smooth muscle cells through regulating PTEN/PI3K/AKT pathway.

OBJECTIVE: Aberrant microRNAs (miRNAs) play vital roles in various human diseases, including atherosclerosis (AS). MiR-647 expression was highly elevated in AS samples. Therefore, this study aimed at exploring the role and mechanism of miR-647 on AS progression. PATIENTS AND METHODS: Human aorta vascular smooth muscle cells (HA-VSMCs) were treated with oxidized modified low-density lipoprotein (ox-LDL) to establish the AS model in vitro. The qRT-PCR assay was used to detect the expression of miR-647 and PTEN mRNA. The levels of PTEN protein, PI3K, AKT, p-PI3K, and p-AKT were measured using Western blot. Cell proliferation and migration were determined by Cell Counting Kit-8 (CCK-8) assay and transwell assay, respectively. The target of miR-647 was verified using the dual-luciferase reporter assay. RESULTS: Our data supported that miR-647 was upregulated and PTEN was downregulated in the serum of AS patients and ox-LDL-treated HA-VSMCs. The proliferation and migration of ox-LDL-treated HA-VSMCs were promoted by miR-647 overexpression or PTEN knockdown, while they were suppressed following miR-647 depletion or high PTEN expression. Moreover, PTEN was a direct target of miR-647. PTEN antagonized miR-647-mediated regulatory effects on cell proliferation and migration. Additionally, the PI3K/AKT signaling pathway was involved in miR-647/PTEN-mediated regulation in ox-LDL-treated HA-VSMCs. CONCLUSIONS: MiR-647 promoted the proliferation and migration of ox-LDL-treated HA-VSMCs at least partly by targeting the PTEN/PI3K/AKT pathway. Targeting miR-647 may be a promising method for AS treatment.

[Proportion and role of CD45+ erythroid progenitor cells in patients with tongue cancer metastasis].

Objective: To investigate the proportion and role of CD45+ erythroid progenitor cells (EPC) in patients with tongue cancer metastasis. Methods: The initial treatment of tongue cancer patients (n=40) from January 2017 to June 2018 in He'nan Provincial People's Hospital was included in this study. According to the presence or absence of lymph node metastasis, they were divided into tumor group (no lymph node metastasis was found in imaging and pathology) and metastasis group (both imaging and pathology confirmed lymph node metastasis). The expression of Ki-67 was detected by immunohistochemistry and the proportion of CD45+CD71+TER119+EPC was detected by flow cytometry. EPC was sorted by flow cytometry, interleukin-10 (IL-10) and transforming growth factor-ß (TGF-ß) were detected by enzyme-linked immunosorbent assay (ELISA), and reactive oxygen species (ROS) was detected by flow cytometry. Transwell was used for tumor invasion test; methyl thiazolyltetrazolium (MTT) assay was used to detect proliferation level. Results: There were 20 cases in the tumor group and metastasis group. There was no significant difference between the two groups in terms of age, sex, time of onset and size of tumors. Flow cytometry showed that the ratio of CD45+EPC in peripheral blood of tumor group and metastasis group was (1.2±0.2)% and (3.1±0.2)% (t=7.823, P<0.001). Correlation analysis showed that the ratio of CD45+EPC was positively correlated with the proliferation index of Ki-67 cells (r=0.592, P=0.006). The results of flow cytometry showed that the mean fluorescence intensity (MFI) of ROS in EPC was 102.1±22.9 in tumor group and 530.0±67.2 in metastasis group (t=6.025,P<0.001). The results of ELISA showed that the mass concentrations of IL-10 and TGF-ß in EPC supernatant of tumor group were (10.8±1.6) and (3.2±0.8) µg/L, respectively. The mass concentrations of IL-10 and TGF-beta in EPC supernatant of metastasis group were (26.9±3.7) and (6.4±0.9) µg/L, respectively (t=3.956, P=0.003; t=2.595, P=0.027). Transwell results showed that the proportion of invasive cells in the CD45+EPC group [(40.3±4.4)%] was higher than that in the control group [(17.5±2.2)%] (t=4.607, P=0.001). MTT proliferation experiment showed that the proliferation rate of the CD45+EPC group [(52.0±3.3)%] was higher than that of the control group [(30.5±1.9)%] (t=5.656, P<0.001). Conclusions: The proportion of CD45+EPC in patients with tongue cancer metastasis is significantly increased. CD45+EPC can promote the proliferation and metastasis of tongue cancer by secreting immunosuppressive molecules and ROS.

Overexpression of lncRNA TCL6 promotes preeclampsia progression by regulating PTEN.

OBJECTIVE: The aim of this study was to explore the role of long non-coding RNA (lncRNA) TCL6 in preeclampsia (PE) development and to investigate its underlying mechanism. PATIENTS AND METHODS: The expression of TCL6 in 42 placental tissues of PE pregnancies and normal pregnancies was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Receiver Operating Characteristic (ROC) curve was applied to explore the relationship between TCL6 expression, urine protein level, blood pressure and neonatal weight of PE pregnancies. The proliferation and cell cycle of trophoblast cells after TCL6 knockdown were detected by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Moreover, the specific role of TCL6 in cell cycle was detected by Western blot. RESULTS: TCL6 was highly expressed in 42 placental tissues of PE pregnancies when compared with that of normal pregnancies. PE pregnancies with lower expression level of TCL6 exhibited significantly lower urinary protein level, as well as systolic and diastolic blood pressure than those with higher level. Besides, neonatal weight was significantly higher in PE pregnancies with lower expression level of TCL6. Meanwhile, downregulation of TCL6 resulted in remarkably increased proliferation and cell cycle of trophoblast cells. In addition, Western blot results indicated that TCL6 knockdown significantly upregulated CDK2 and downregulated PTEN in trophoblast cells. CONCLUSIONS: TCL6 was highly expressed in placental tissues of PE patients. Overexpression of lncRNA TCL6 inhibited the proliferation of trophoblast cells and promoted PE development via targeting PTEN.

Research on the correlation of changes in plasma lncRNA MEG3 with change in inflammatory factors and prognosis in patients with traumatic brain injury.

OBJECTIVE: To study the correlation between the plasma long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3) and the levels of inflammatory cytokines in patients with traumatic brain injury (TBI) and to evaluate its prognosis to screen new biological targets for the diagnosis and treatment of TBI. PATIENTS AND METHODS: 40 patients with TBI (TBI group) and 40 healthy people (control group) were collected and venous blood was drawn. The plasma MEG3 in subjects was quantitatively analyzed via quantitative Polymerase Chain Reaction (qPCR). Moreover, the levels of inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, and IL-8] in plasma in each group were detected via enzyme-linked immunosorbent assay (ELISA). Finally, the correlation analysis was performed for the MEG3 expression level and inflammatory cytokine levels in patients with TBI. Patients were divided into high-expression MEG3 group and low-expression MEG3 group, high-level inflammatory cytokine group and low-level inflammatory cytokine group according to the median, followed by prognosis evaluation. The MEG3 expression level in TBI group was significantly decreased compared to that in control group, and the levels of inflammatory cytokines in plasma, including TNF-α, IL-1ß, IL-6, and IL-8, were significantly higher than in control group. RESULTS: The results of the correlation analysis showed that the expression level of plasma MEG3 had a significantly negative correlation with the level of each inflammatory cytokine. The prognostic analysis revealed that the prognosis of patients with high MEG3 expression level and low inflammatory cytokine levels was good, while it was poor in patients with low MEG3 expression level and high inflammatory cytokine levels; the difference was significant. In patients with TBI, the expression level of plasma MEG3 is decreased, while the inflammatory cytokine levels are increased, and there is a significantly negative correlation between the two items. CONCLUSIONS: The prognosis of patients with high MEG3 expression level and low inflammatory cytokine levels is good so MEG3 and inflammatory cytokines can be used as biomarkers for diagnosis and treatment of TBI, improving the prognosis of patients.

Retraction Note: Structural simulation of adenosine phosphate via plumbagin and zoledronic acid competitively targets JNK/Erk to synergistically attenuate osteoclastogenesis in a breast cancer model.

This article has been retracted at the request of the authors. After publication, the authors found that in Figure 2B-a the first two images in the third row partly overlapped and that there is also overlap between the fourth and fifth image in the second row. The two images were taken from two adjacent wells, treated by ZA 0.3uM-CM or ZA 0.75uM-CM, with or without PL 1.25uM. This overlap may have been caused by mishandling in the imaging process when the authors made microscope observations and so the findings are no longer reliable. All authors agree to this retraction.

[Study on analgesic effect and mechanism of cinobufagin on rats with bone cancer pain].

Objective: To evaluate the analgesic effects of cinobufagin (CBG) on cancer-induced bone pain in rat and study the role of the muscarinic receptor M4 subtype (M4 mAChR) in its involvement. Methods: A total of 100 Female Sprague-Dawley rats were randomly divided into 5 groups (n=20): Sham group (group S), Cancer group (group A), Normal saline + CBG vehicle solution group (group ANS), Normal saline + 1 mg/kg CBG group (group ANC) and Tropicamide + 1 mg/kg CBG group (group ATC). Rats in group S were injected 10 µl Hank's solution into the left tibia medullar cavity, while rats in group A, ANS, ANC, and ATC were injected Walker 256 mammary cancer cells (10 µl, 2×10(7) cells/ml) into the same place. On day 9 post-inoculation rats in group ANS, ANC, and ATC were respectively received Saline (0.9%, 15 µl, i.t.), Saline (0.9%, 15 µl, i.t.)and 10 nmol of M4 mAChR blocker Tropicamide. After 10 min, ANS group, ANC group and ATC group were intraperitoneally injected with CBG vehicle solution, 1 mg/kg CBG and 1 mg/kg CBG. Model rats in each group were tested three times average as its basis pain threshold before injection cancer cells (T(0)). Mechanical withdrawal thresholds were measured on left hind paws, before 20 min (T(1)) and after 10 min (T(2)), 30 min (T(3)), 60 min (T(4)), 90 min (T(5)) and 120 min (T(6)) intrathecal injection. Left L4-L6 spinal dorsal horn and DRG were removed for determination of the expression of CaM-dependent kinaseⅡa (CaMKⅡa) and pCaMKⅡa by Western Blot after 60 min drug delivery. Results: At each time point from T(1) to T(6), the mechanical pain thresholds of group S were (8.69±0.45), (8.63±0.44), (8.65±0.39), (8.84±0.23), (8.80±0.14), (8.75±0.14) g, respectively, and the mechanical pain thresholds of group A were (6.37±0.30), (6.42±0.13), (6.29±0.17), (6.25±0.22), (6.34±0.33), (6.36±0.34) g, the difference was statistically significant (t=-16.41, -23.47, -30.25, -17.35, -19.52, -22.56, all P<0.01). At each time point from T(3) to T(5), the mechanical pain thresholds of the ANS group were (6.42±0.32), (6.39±0.34), (6.26±0.32) g, respectively, and the mechanical pain thresholds of the ANC group were (7.29±0.34), (7.81±0.15), (7.54±0.19) g, the difference was statistically significant (t=13.52, 14.22, 17.33, all P<0.01). At each time point from T(3) to T(5), compared with the ANC group, the mechanical pain threshold of the ATC group decreased (6.55±0.23), (6.84±0.46), (6.80±0.43) g, and the difference was statistically significant (t=-12.69, -11.26, -10.33, all P<0.01). At the time of T(4), the expressions of pCaMKⅡa in the spinal dorsal horn of each group were (0.67±0.05), (1.64±0.12), (1.57±0.14), (0.78±0.09), (1.39±0.11), respectively, and the expressions of pCaMKⅡa in DRG of each group were (1.65±0.39), (3.59±0.17), (3.43±0.32), (2.17±0.34), (2.95±0.23). The differences were statistically significant (F=179.89, 198.76, both P<0.01). Compared with the S group, the expression of pCaMⅡa was up-regulated in group A. Compared with ANS group, the expression of pCaMKⅡ a was down-regulated in ANC group. Compared with ANC group, the expression of pCaMK Ⅱ a was up-regulated in ATC group. The expression of CaMKⅡa in spinal dorsal horn and DRG was not statistically significant (F=1.25, 2.79, both P>0.05). Conclusions: These results demonstrated that M4mAChR participated in mediating the alleviation of hyperalgesia by cinobufagin in rats with bone cancer pain, and its mechanism may be related to pCaMKⅡa/CaMKⅡa signaling pathway.

[The pathological characteristics and outcomes of 40 surgically treated stage M1b non-small cell lung cancer patients].

Objective: Clinical evidences of surgically treated stage M1b non-small cell lung cancer (NSCLC) patients were limited. This study aimed to summarize the clinical data of these patients to explore the prognostic factors of this population. Methods: From January 1999 to December 2012, the clinical data of 40 stage M1b NSCLC patients, including 24 males and 16 females, who underwent surgery were collected by Cancer Hospital, Chinese Academy of Medical Sciences. Kaplan-Meier method, log rank test and Cox risk regression model were used to analyze the prognose of these patients and their influence factors. Results: A total of 40 patients were available for survival analysis. The Survival rates of the whole population at 1, 3 and 5 years were 70.0%, 40.0% and 22.5%, respectively. The median survival time (MST) was 31.5 months. The outcomes of 23 patients underwent primary tumor resection and local treatment of metastatic site (MST: 41.5 months) were significantly better than those of 13 patients with only primary tumor resection (MST: 15.5 months) and 4 patients with thoracic exploration (MST: 14.5 months) (P<0.05). Multivariate analysis showed that patients without pleural effusion, brain metastasis, chemotherapy and targeted therapy had a statistically better survival (P<0.05). Conclusions: The overall survivals of surgically treated stage M1b NSCLC patients appear encouraging, and some selected patients may even achieve a long-time survival. Multimodality treatment including surgical lung resection and radical treatment of metastasis should be considered for these patients.

[Analyses of pathogenic factors and clinicopathological characteristics of vocal leukoplakia].

Objective: To investigate the pathogenic factors of vocal leukoplakia and its clinical and pathological features. Methods: Eighty-one patients with vocal cord leukoplakia who underwent surgery between February 2010 and December 2016 and 160 volunteers without pharyngeal symptoms designed as controls were included in this case control study. The clinicopathological characteristics of 81 patients were summarized and analyzed synthetically. Results: There was statistical significance in reflux symptom index(RSI), reflux finding score(RFS), smoking index, and drinking index between case group and control group(Z=-5.35, -4.82, -4.76, -2.44, P<0.05). The voice-using duration per day in case group was significantly longer than that of control group.There was no statistical significance in hospital anxiety and depression scale for anxiety(HADA) scores、hospital anxiety and depression scale for depression(HADD) scores between case group and control group(P>0.05). In 42 patients who received 24-hour dual probe pH monitoring the prevalence of pathologic LPR was 42.8%. In 81 patients, 39(48%)patients were pathologically diagnosed as squamous cell hyperplasia, 18(22%)patients as mild dysplasia, 12(15%)sides as moderate dysplasia , 10(12%)patients as severe dysplasia and 2(2%)patients as carcinoma in-situ. The average age of high-risk pathological vocal leukoplakia was significantly higher than that of low-risk leukoplakia(t=-2.73, P<0.01). The propotion of speckled leukoplakia in high-risk leukoplakia was significantly higher than that of low-risk leukoplakia(χ(2)=23.81, P<0.01). There was no statistical significance between high-risk leukoplakia and low-risk leukoplakia in the prevalence of pathologic LPR(P>0.05). The bilateral lesions, speckled leukoplakia were more likely to relapse(χ(2)=4.27, 12.17, P<0.05). The more serious the pathology, the more likely it was to relapse (Z=-2.168, P=0.03). There was no statistical significance between recurrence group and non-recurrence group in the prevalence of pathologic LPR(P>0.05). Conclusions: LPR, smoke constitute the risk factors of vocal cord leukoplakia. Drinking, voice abuse are related to vocal cord leukoplakia. Senile, speckled leukoplakia are more likely to be malignancy. A speckled leukoplakia, bilateral leukoplakia, severe pathological degree are important factors to predict recurrence.

[Analysis of clinical characteristics and prognosis of neuromyelitis optic spectrum disease complicated with malignant tumor].

Objective: To investigate the clinical features of patients involved with both malignant tumors and neuromyelitis optica spectrum disorders (NMOSD). Methods: The clinical data of 473 patients with NMOSD admitted into the Third Affiliated Hospital of Sun Yat-sen University and the Department of Neurology, Affiliated Hospital of Guizhou Medical University from June 2012 to March 2017 were retrospectively analyzed. Eleven NMOSD patients complicated with malignant tumors were screened out (3 with breast cancer, 2 with cervical cancer, 2 with rectal cancer, 2 with leukemia, 1 with nasopharyngeal carcinoma, 1 with thyroid cancer). Fifty patients without NMOSD were included as controls. Results: Most of the NMOSD patients were complicated by low-moderately differentiated squamous cell carcinoma/adenocarcinoma, mainly seen in breast, reproductive system, digestive system and hematological system. In terms of sex ratio and autoantibodies, the NMOSD patients with and without malignant tumors showed no significant difference. However, comparing to the patients without malignant tumor, the ones with malignant tumor showed a tendency of lower rate of initial brain symptoms and relapse rate, while with older onset age, higher initial EDSS score, protein content in cerebrospinal fluid (CSF), higher rates of initial symptom resulted from the focus of posterior region of the medulla and of significant image focus. Of the 8 NMOSD patients who diagnosed as malignant tumors in our hospital, 2 with breast cancer and 1 with cervical cancer had a good prognosis (follow-up EDSS score <3). All the 3 patients received aggressive surgery and chemotherapy treatment. However, the other 5 patients had poor prognosis (follow-up EDSS score ≥3 points). All the 11 patients received anti-tumor therapy, 4 patients had first NMOSD attack after anti-tumor treatment and no relapse. Only one case from the remaining 7 patients had relapse; Among the 9 patients received immunosuppressive therapy, 7 patients had no relapse, and 8 cases maintained stable; while, among all the 9 patients received immunosuppressive agents and anti-tumor therapy, only one case had relapse. Conclusions: There are some differences in the clinical features between the NMOSD patients with malignant tumors and the NMOSD patients without malignant tumors. Immunosuppressive therapy can improve the prognosis of patients with NMOSD and tumor, without increasing the risk of malignant tumor. The pathological type, staging and antitumor therapy may influence the prognosis of NMOSD. NMOSD patients with malignant tumor could be treated with anti-tumor and immunosuppressive agents if needed.