ObjectiveTo explore the role of interleukin-6 （IL-6）/signal transducer and activator of transcription 3 （STAT3） pathway in the balance of T helper 17 （Th17）/regulatory T （Treg） cells in ulcerative colitis （UC） with internal dampness-heat accumulation syndrome and the intervention mechanism of Shaoyaotang. MethodA total of 60 SD rats were randomized into blank group （equivalent volume of normal saline）， model group （equivalent volume of normal saline）， western medicine control group （0.42 g·kg^-1 mesalazine）， and low-dose （11.1 g·kg^-1）， medium-dose （22.2 g·kg^-1）， and high-dose （44.4 g·kg^-1） Shaoyaotang groups. UC with internal dampness-heat accumulation syndrome was induced in rats with the compound method except for the blank group. The administration lasted 14 days for each group. At 24 h after the last administration， rats were killed and the spleen and colon tissues were separated. The histopathological changes of colon were observed based on hematoxylin and eosin （HE） staining and the levels of interleukin-17 （IL-17） and transforming growth factor-β₁ （TGF-β₁） in colon tissue were detected by immunohistochemistry （IHC）. Flow cytometry was employed to determine the levels of Th17/Treg cells in the spleen， and Western blot to measure the levels of IL-6 and STAT3 proteins in colon tissue. ResultCompared with the blank group， the model group had lesions such as congestion and erosion， low percentage of spleen Treg cells （P<0.01）， high percentage of Th17 cells （P<0.01）， and high levels of IL-6 and STAT3 proteins in colon tissue （P<0.01）. Compared with the model group， the administration groups showed alleviation of colon injury， high percentage of spleen Treg cells （P<0.05， P<0.01）， low percentage of Th17 cells （P<0.01）， and low levels of IL-6 and STAT3 proteins in colon tissue （P<0.01）. ConclusionShaoyaotang regulates the balance of Th17/Treg by inhibiting the IL-6/STAT3 pathway， thereby relieving the pathological damage of UC rats with internal dampness-heat accumulation syndrome and affecting their immune function.

BACKGROUND: Anlotinib is a newly developed small molecule multiple receptor tyrosine kinase (RTK) inhibitor that was approved for the treatment of patients with lung cancer in China. We aim to report 3 cases of rare complication of anlotinib-bronchial fistula (BF) during the treatment of lung cancer patients and summarize the possible causes. METHODS: We collected three patients who developed BF due to anlotinib treatment, and conducted a search of Medline and PubMed for medical literature published between 2018 and 2020 using the following search terms: "anlotinib," "lung cancer," and "fistula." RESULTS: Our literature search produced two case reports (three patients) which, in addition to our three patients. We collated the patients' clinical characteristics including demographic information, cancer type, imaging features, treatment received, risk factors for anlotinib related BF, and treatment-related outcomes. The six patients shared some common characteristics: advanced age, male, concurrent infection symptoms, diabetes mellitus (DM), advanced squamous cell and small cell lung cancers, centrally located tumors, tumor measuring ≥5 cm in longest diameter, and newly formed tumor cavitation after multi-line treatment especially after receiving radiotherapy. Fistula types included broncho-pericardial fistula, broncho-pleural fistula, and esophago-tracheobronchial fistula. Six patients all died within 6 months. CONCLUSIONS Although anlotinib is relatively safe, it is still necessary to pay attention to the occurrence of BF, a rare treatment side effect that threatens the quality of life and overall survival of patients. Anlotinib, therefore, requires selective use and close observation of high-risk patients.

Objective: To investigate the prevalence of hyperuricemia and its correlation among adolescents aged 13-16 in Hanjiang district of Yangzhou city. Methods: Five hundred and nine adolescents aged 13-16 years in Hangjiang district of Yangzhou city were recruited. Their fasting serum uric acid level were measured by Benecheck premium UA monitoring system. The demographic imformation, family history, eating and living habits of the respondents were collected by questionnaires. Results: The average serum uric acid level of 509 adolescents in Hangjiang district of Yangzhou city was (374±98) μmol/L, and (420±103) μmol/L in males, higher than that of females [(329±70) μmol/L](t=11.69, P<0.05). The overall prevalence of HUA was 36.9%(188/509), and 43.2%(108/250) in males, higher than in females 30.8%(80/259)(χ²=4.16, P<0.05). Multivariate Logistic regression analysis showed that male [OR=1.70, 95%CI(1.18, 2.45)], obese [OR=8.99, 95%CI(4.58, 17.63)], high-salt oily diet [OR=1.67, 95%CI(1.05, 2.61)], soy products [OR=2.66, 95%CI(1.45, 4.88)], animal offal [OR=7.81, 95%CI(3.42, 17.83)], seafood [OR=4.80, 95%CI(2.83, 8.12)], sugary drinks [OR=6.49, 95%CI(3.08, 13.64)] and time spent on electronic products [OR=1.98, 95%CI(1.07, 3.65)] might be risk factors for hyperuricemia (P<0.05), while fruit [OR=0.52, 95%CI(0.32, 0.85)] and outdoor activities [OR=0.55, 95%CI(0.33, 0.92)] might be protective factors for HUA (P<0.05). Conclusion There is a high prevalence of hyperuricemia among adolescents aged 13-16 in Hangjiang district of Yangzhou city. The level of serum uric acid in adolescents significantly increased. Male, obesity, high salt and oily diets, soy products, animal offal, seafood, and sugary drinks may be risk factors for HUA. Fruits and outdoor activities may be protective factors for HUA. Relevant disciplines can take corresponding measures to prevent adolescent HUA.

Objective To study the clinical significance of thromboelastography (TEG) for determining the presence of coagulation disorders in septic children.Methods A total of 100 patients suffering from sepsis or severe sepsis in pediatric intensive care unit (PICU) of Shanghai Children's Medical Center from February 2014 to January 2015 were recruited.TEG tests and conventional coagulation laboratory tests (CCTs) including platelet count,fibrinogen,prothrombin time (PT),activated partial thromboplastin time,D-dimers,and international normalized ratio (INR) were carried out in all patients at the primary diagnosis of sepsis.Another 25 healthy children taking physical examination were enrolled as control group.Rank Sum Test was used to detect the differences in coagulation markers and TEG between the groups and there was statistical significance when P ＜ 0.05.Receiver operating characteristic (ROC) curves were used to evaluate the roles of TEG and CCTs tests in this study.Results Of them,there were 56 patients with sepsis and 44 with severe sepsis.The male to female ratio was 63∶ 37,the median age was 11.5 (3.3-48) months,and 71％ patients suffered from underlying disease.According to TEG,72 patients had coagulation disorders,including 28 with hypercoagulation and 44 with hypocoagulation.CCTs tests showed 50 patients had coagulation disorders,including 29 with non-overt DIC and 21 with overt DIC.The rate of hypercoagulability was significantly higher in non-DIC group than in non-overt DIC group (46％vs.17.2％,P =0.016).The rate of hypocoagulability was significantly higher in overt DIC group than in non-overt DIC group (100％ vs.44.8％,P ＜ 0.01).Patients with hypercoagulation disorders had significantly shorter R (coagulation reaction time) and K (coagulation formation time) and greater α (angle α),MA (maximal amplitude) and CI (comprehensive coagulation index) compared with control group (P ＜ 0.01).According to CCTs results,patients with hypercoagulation had significantly prolonged PT compared with control group (P =0.002).Compared with sepsis group,severe sepsis group had significantly prolonged R and K and lower α,MA and CI (P ＜ 0.01).ROC analysis demonstrated that area under the curve (AUC) of TEG and CCTs variables for diagnosis of severe sepsis were significantly greater than 0.5.Both variables of α (P =0.000 2) and K (P =0.004 1) had significantly greater AUCs compared with Fib.Conclusions There were 72％ septic patients with coagulation disorders.The hypercoagulability occurred earlier in patients with sepsis and the hypocoagulability occurred later in patients with severe sepsis.The TEG may provide important information for clinicians to deal with coagulation disorders in septic children.

Objective To investigate the role and mechanism of miR-155 in invasion and metastasis of lung adenocarcinoma A549 cells.Methods Real-time PCR and fluorescence in situ hybridization were used to detect the miR-155 expression in patients′lung adenocarcinoma and adjacent tissue and lymph nodes.Scratch test and Transwell migration assay were used to assess the effect of miR-155 on the A549 cell migration and invasion capability.Bioinformatics software was used to predict the target genes of miR-155, and using luciferase to assay the target gene.Western blot and real-time PCR were performed to confirm the role of miR-155 expression in the regulation of target gene PTEN.Results The real-time quantitative PCR showed that the miR-155 expression levels in adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were 4.1±0.5, 9.6±3.1 and 7.8±2.2, respectively.The in situ hybridization showed that the expression rates of miR-155 in the adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were (23.2±15.3)%, (75.4±20.2 )% and (60.4±25.1)%,respectively.The Scratch assay showed that the wound healing rates in the miR-155 mimics group, miR-155 mimics NC group, miR-155 inhibitor group and miR-155 inhibitor NC group at 24 h were (43.2±2.2)%, (21.3±4.2)%, (24.3±5.3)%, and (35.2± 5.1)%, and that at 48 h were (75.2±4.5)%, (52.6±5.2)%, (39.4±4.2)%, and( 51.5±4.3)%, respectively.Dual luciferase reporter gene assay showed that the value of the luciferase in the miR -155 mimics group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 4.7±0.5 and 7.3±0.7, and the miR-155 mimics luciferase values of the control group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 7.8±0.9 and 7.5±0.8, respectively.The real-time quantitative fluorescence PCR showed that the relative expression of PTEN protein in the miR-155 mimics group, miR-155 mimics control group, miR-155 mimics inhibitor group, and miR-155 inhibitor control group were 0.5± 0.3, 1.0±0.1, 2.2±0.2 and 1.2 ±0.1, respectively.The Western blot assay detected that the relative expression of PTEN protein levels in the miR-155 mimics group, miR-155 mimics control group, miR-155 inhibitor group and miR-155 inhibitor control group were 0.4±0.1, 1.0±0.3, 2.8±0.2 and 1.4 ±0.1, respectively.The differences in PTEN mRNA and protein expressions of the four groups were statistically significant (P<0.05 for all).Conclusions miR-155 may promote the invasion and metastasis of lung adenocarcinoma through reducing the target PTEN gene expression.

Objective To investigate the role and mechanism of miR-155 in invasion and metastasis of lung adenocarcinoma A549 cells.Methods Real-time PCR and fluorescence in situ hybridization were used to detect the miR-155 expression in patients′lung adenocarcinoma and adjacent tissue and lymph nodes.Scratch test and Transwell migration assay were used to assess the effect of miR-155 on the A549 cell migration and invasion capability.Bioinformatics software was used to predict the target genes of miR-155, and using luciferase to assay the target gene.Western blot and real-time PCR were performed to confirm the role of miR-155 expression in the regulation of target gene PTEN.Results The real-time quantitative PCR showed that the miR-155 expression levels in adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were 4.1±0.5, 9.6±3.1 and 7.8±2.2, respectively.The in situ hybridization showed that the expression rates of miR-155 in the adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were (23.2±15.3)%, (75.4±20.2 )% and (60.4±25.1)%,respectively.The Scratch assay showed that the wound healing rates in the miR-155 mimics group, miR-155 mimics NC group, miR-155 inhibitor group and miR-155 inhibitor NC group at 24 h were (43.2±2.2)%, (21.3±4.2)%, (24.3±5.3)%, and (35.2± 5.1)%, and that at 48 h were (75.2±4.5)%, (52.6±5.2)%, (39.4±4.2)%, and( 51.5±4.3)%, respectively.Dual luciferase reporter gene assay showed that the value of the luciferase in the miR -155 mimics group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 4.7±0.5 and 7.3±0.7, and the miR-155 mimics luciferase values of the control group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 7.8±0.9 and 7.5±0.8, respectively.The real-time quantitative fluorescence PCR showed that the relative expression of PTEN protein in the miR-155 mimics group, miR-155 mimics control group, miR-155 mimics inhibitor group, and miR-155 inhibitor control group were 0.5± 0.3, 1.0±0.1, 2.2±0.2 and 1.2 ±0.1, respectively.The Western blot assay detected that the relative expression of PTEN protein levels in the miR-155 mimics group, miR-155 mimics control group, miR-155 inhibitor group and miR-155 inhibitor control group were 0.4±0.1, 1.0±0.3, 2.8±0.2 and 1.4 ±0.1, respectively.The differences in PTEN mRNA and protein expressions of the four groups were statistically significant (P<0.05 for all).Conclusions miR-155 may promote the invasion and metastasis of lung adenocarcinoma through reducing the target PTEN gene expression.

Background and objective Pulmonary sarcomatoid carcinoma is a rare histologic subtype of non-small cell lung cancer. hTe effective treatment for this disease has not well deifned due to its extremely low morbidity. hTis study explores the clinicopathological characteristics and prognosis of 38 patients with PSC, so as to provide some clues for its diagnosis and treatment. Methods hTe study enrolled 38 patients with PSC that were diagnosed with histology or cytology in our hospital between January 2000 and December 2013. We retrospectively analyzed general clinical characteristics, smoking history, tumor size, TNM staging, pathology, immunohistochemistry, diagnostic method, treatment and prognosis. We used SPSS 19.0 statistical sotfware and Kaplan-Meier method to analyze our data. Results Patients in this study were aged from 26 to 76 years old (the median age was 57.5 years old). Among all of them, the male to female ratio was 4:1, and 81.6%of patients had smoking history. Cough and hemoptysis were the most common primary symptoms. hTe median survival was 21 months, while one-year survival rate, three-year survival rate and five-year survival rate were 68.4%, 31.6%and 18.4%respectively. Tumor size, TNM staging, distant metastasis and surgery therapy were associated with the prognosis of patients. Conclusion Patients with PSC present with no special symptoms generally. According to our study, factors that affect patients’ prognosis include tumor size, TNM staging, distant metastasis and surgery. Complete resection is the key treatment for PSC patients, but comprehensive chemoradiotherapy needs further exploration in evidence-based medicine. Biological target therapy may give new insight into treatment for PSC.

BACKGROUNDTumor cells can reduce the number of dendritic cells (DCs) in the tumor environment and cause DC dysfunction through autocrine or paracrine pathways. We sought to measure cyclooxygenase-2 (COX-2) expression in bombesin-inhibited DCs treated with theanine in vitro and to explore the protection and activation effects of theanine on DCs.

METHODSEnzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were used to analyze the effects of theanine on COX-2 expression and interleukin (IL)-12/IL-10 secretion of bombesin-treated DCs.

RESULTSDCs acquired an impaired phenotype as a result of bombesin treatment. Theanine increased the expression of mature DC surface molecules. The number of cell apoptosis with the treatment of bombesin and theanine significantly decreased, accounting for 15.9%, compared with 26.1% of cell apoptosis with bombesin. COX-2 expression in bombesin-treated DCs was inhibited by theanine in a dose-dependent manner. Theanine promoted DC secretion of IL-12. IL-12 levels reached (137.4 ± 4.9) pg/ml with theanine at 200 µmol/L. However, theanine inhibited the secretion of IL-10 in a dose-dependent manner. IL-10 levels were only (58.4 ± 6.9) pg/ml with theanine at 200 µmol/L.

CONCLUSIONTheanine inhibits the transcription and translation of COX-2 and regulates the balance of IL-10/IL-12 secretion in bombesin-inhibited DCs, leading to the recovery of a state of activation in DCs.

Bombesin ; pharmacology ; Cells, Cultured ; Cyclooxygenase 2 ; metabolism ; Dendritic Cells ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Glutamates ; pharmacology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism

Objective To establish the triplex Taqman probes real-time RT-PCR method for simultaneously detecting of EV71,CA16 and EV.Methods Retrospective study.Specific primers and probes were designed based on conserved regions of EV71,CA16 and EV.The sensitivity,specificity and reproducibility were assessed by the optimized reaction system.A total of 176 throat swabs as the experimental group were collected from children with suspected hand foot mouth disease (HFMD),who admitted from April 2012 to July 2012 in Nanjing Children's Hospital affiliated to Nanjing Medical University.During this time,10 cases of healthy children,10 cases of outpatients with flu-like symptoms and 90 cases of inpatients in pneumology department of our hospital were recruited as control group,whose throat swabs were also collected.All of 286 samples were tested by the triplex Taqman probes real-time RT-PCR for simultaneously detecting EV71,CA16 and EV.SPSS13.0 was used to analyze the results.Results The sensitivities of the triplex Taqman probes real-time RT-PCR was 1.0 × 103 copies per milliliter for EV71,CA16 and EV.It showed 100％ specificity for 9 enterovirus and 3 non-enterovirus.Analysis with 1.0 × 103-1.0 × 105 copies per milliliter constructed plasmids demonstrated high reproducibility with coefficient of variation of 0.44％-1.04％ for EV71,0.38％-0.73％ for CA16,and 0.46％-0.90％ for EV.More over 176 samples collected from children with suspected HFMD were detected by triplex Taqman probes real-time RTPCR and real-time RT-PCR.The results showed 97.2％ (171/176)agreement and 0.94 Kappa value with high concordance.Conclusions The triplex Taqman probes real-time RT-PCR detecting EV71,CA16 and EV simultaneously has been established successfully.The assay,with high sensitivity and specificity,provide good basis for the rapid clinical diagnosis of EV71,CA16 and EV and open up broad prospects for clinical and relevant researches.

ObjectiveTo investigate the prognostic factors of non-small cell lung cancer (NSCLC) after resection.MethodsClinical data of 131 NSCLC patients who underwent resection were reviewed and divided into chemotherapy group(86 cases) and non-chemotherapy group(45 cases) according to the treatment method.Survival rate was calculated by Kaplan-Meier method.The prognosis was analyzed by Cox proportional hazards model.ResultsThe median survival time (MST) of squamous cell carcinoma (76 cases),bronchial alveolar cell carcinoma( 8 cases),adenocarcinoma( 35 cases ),adenosquamous carcinoma (12 cases) was 60,54,34,24 months respectively (P＜0.05).For the patients of stage Ⅰ B,the MST of chemotherapy group and non-chemotherapy group was 75 and 76 months respectively(P ＞ 0.05 ).Multivariate analysis showed that tumor size,T stage,N stage,chemotherapy were independent prognostic factors (P =0.080,0.002,0.000,0.029).Conclusions Squamous cell carcinoma and bronchial alveolar cell carcinoma have better prognosis than adenocarcinoma,adenosquamous carcinoma.For the patients of stage Ⅰ B,the survival time can't be prolonged through platinum-based chemotherapy.Tumor size,T stage,N stage,chemotherapy are independent prognostic factors.