Etiology and clinical features of children with bronchiectasis in China: A 10-year multicenter retrospective study.

INTRODUCTION: The current study aims to investigate the etiology spectrum and the clinical characteristics of bronchiectasis in Chinese children. METHODS: The study is designed as a multicenter retrospective study. 193 cases were enrolled in 13 centers in China between 2008 and 2017. The inclusive cases must meet the clinical as well as the HRCT criteria. Only if both two radiologists confirmed the diagnosis, the case could be enrolled. The cases that could not provide clinical and imageology data were excluded. The data were entered into the specialized system and then analyzed. RESULTS: One hundred sixty-nine cases (87%) were found to have the underlying etiology. Post-infective (46%), primary immunodeficiency (14%), and PCD (13%) were the common causes. All cases came from 28 provinces in Mainland China. The median age of symptom onset was 5.8 (2.0, 8.9) years. The median age of diagnosis was 8.4 (4.5, 11.6) years. The main symptoms were cough, sputum expectoration, and fever during the exacerbation. Nineteen percent of patients suffered from limited exercise tolerance. Clubbing was found in 17% of cases. Nearly 30% of patients presented growth limitations. On the HRCT findings, 126 cases had diffused bronchiectasis, and bilateral involvement was found in 94 cases. The lower lobes and right middle lobes were most commonly involved. Approximately 30% of cultures of sputum and bronchoalveolar lavage were positive. CONCLUSION: A majority of cases could be found the underlying etiology. Post-infective, primary immunodeficiency, and PCD were the most common causes. Some clinical figures might indicate a specific etiology.

Clinical Analysis of Primary Tracheobronchial Tumors in Children and Evaluation of the Predicting Models for Mucoepidermoid Carcinoma.

OBJECTIVE: To determine the clinical characteristics and prognosis of primary tracheobronchial tumors (PTTs) in children, and to explore the most common tumor identification methods. METHODS: The medical records of children with PTTs who were hospitalized at the Children's Hospital of Chongqing Medical University from January 1995 to January 2020 were reviewed retrospectively. The clinical features, imaging, treatments, and outcomes of these patients were statistically analyzed. Machine learning techniques such as Gaussian naïve Bayes, support vector machine (SVM) and decision tree models were used to identify mucoepidermoid carcinoma (ME). RESULTS: A total of 16 children were hospitalized with PTTs during the study period. This included 5 (31.3%) children with ME, 3 (18.8%) children with inflammatory myofibroblastic tumors (IMT), 2 children (12.5%) with sarcomas, 2 (12.5%) children with papillomatosis and 1 child (6.3%) each with carcinoid carcinoma, adenoid cystic carcinoma (ACC), hemangioma, and schwannoma, respectively. ME was the most common tumor type and amongst the 3 ME recognition methods, the SVM model showed the best performance. The main clinical symptoms of PPTs were cough (81.3%), breathlessness (50%), wheezing (43.8%), progressive dyspnea (37.5%), hemoptysis (37.5%), and fever (25%). Of the 16 patients, 7 were treated with surgery, 8 underwent bronchoscopic tumor resection, and 1 child died. Of the 11 other children, 3 experienced recurrence, and the last 8 remained disease-free. No deaths were observed during the follow-up period. CONCLUSION: PTT are very rare in children and the highest percentage of cases is due to ME. The SVM model was highly accurate in identifying ME. Chest CT and bronchoscopy can effectively diagnose PTTs. Surgery and bronchoscopic intervention can both achieve good clinical results and the prognosis of the 11 children that were followed up was good.

[Clinical and microbiological characteristics of children with drowning-associated aspiration pneumonia]. / æººæ°´åŽå¸å ¥æ€§è‚ºç‚Žæ‚£å„¿çš„ä¸´åºŠç‰¹å¾åŠç— åŽŸåˆ†æž.

OBJECTIVES: To study the clinical and microbiological characteristics of children with drowning-associated aspiration pneumonia, so as to provide a reference for empirical selection of antibacterial agents. METHODS: A retrospective analysis was performed on the medical data of 185 children with drowning-associated aspiration pneumonia who were admitted to Children's Hospital of Chongqing Medical University from January 2010 to October 2020. According to the drowning environment, these children were divided into four groups: fecal group (n=44), freshwater group (n=69), swimming pool group (n=41), and contaminant water group (n=31). The clinical characteristics and pathogen detection results were reviewed and compared among the four groups. RESULTS: The 185 children had an age of 4 months to 17 years (median 34 months). Sputum cultures were performed on 157 children, and 103 were tested positive (65.6%), with 87 strains of Gram-negative bacteria (68.5%), 37 strains of Gram-positive bacteria (29.1%), and 3 strains of fungi (2.4%). Gram-negative bacteria were the main pathogen in the fecal group and the contaminant water group, accounting for 88.2% (30/34) and 78.3% (18/23), respectively. The freshwater group had a significantly higher detection rate of Gram-positive bacteria than the fecal group (P<0.008), and the swimming pool group had an equal detection rate of Gram-negative bacteria and Gram-positive bacteria. CONCLUSIONS: For pulmonary bacterial infection in children with drowning in feces or contaminant water, antibiotics against Gram-negative bacteria may be applied empirically, while for children with drowning in a swimming pool or freshwater, broad-spectrum antibiotics may be used as initial treatment, and subsequently the application of antibiotics may be adjusted according to the results of the drug sensitivity test.

Consistency between nasopharyngeal aspirates and bronchoalveolar lavage fluid in pathogen detection in children with pneumonia: an analysis of 533 cases. / 533ä¾‹è‚ºç‚Žå„¿ç«¥é¼»å’½æŠ½å¸ç‰©å’Œæ”¯æ°”ç®¡è‚ºæ³¡çŒæ´—æ¶²çš„ç— åŽŸæ£€å‡ºä¸€è‡´æ€§åˆ†æž.

OBJECTIVES: To study the consistency between nasopharyngeal aspirates (NPA) and bronchoalveolar lavage fluid (BALF) in pathogen detection in children with pneumonia. METHODS: A retrospective analysis was performed on the data of pathogens detected in 533 children with pneumonia from February 2017 to March 2020. The paired McNemar's test was used to compare the difference in pathogen detection between NPA and BALF groups. The Kappa coefficient was used to analyze the consistency in pathogen detection between the two groups. RESULTS: NPA had a sensitivity of 28%, a specificity of 74%, a positive predictive value of 14%, and a negative predictive value of 91% in detecting bacteria, and a Kappa coefficient of 0.013 suggested poor consistency between NPA and BALF. NPA had a sensitivity of 52%, a specificity of 81%, a positive predictive value of 24%, and a negative predictive value of 94% in detecting viruses, and a Kappa coefficient of 0.213 suggested poor consistency between NPA and BALF. NPA had a sensitivity of 78%, a specificity of 71%, a positive predictive value of 49%, and a negative predictive value of 90% in detecting Mycoplasma pneumoniae, and a Kappa coefficient of 0.407 suggested moderate consistency between NPA and BALF. CONCLUSIONS: There is poor consistency between NPA and BALF in the detection of bacteria and viruses, and clinicians should be cautious in diagnosing lower respiratory tract infection based on bacteria or viruses detected in NPA. There is moderate consistency between NPA and BALF in the detection of Mycoplasma pneumoniae, suggesting that it may be reliable to diagnose lower respiratory tract infection based on Mycoplasma pneumoniae detected in NPA, while comprehensive judgment in combination with clinical conditions is needed.

[Influencing factors for the clinical effect of bronchoalveolar lavage in children with Mycoplasma pneumoniae pneumonia and atelectasis].

OBJECTIVE: To study the influencing factors for the clinical effect of bronchoalveolar lavage (BAL) in children with Mycoplasma pneumoniae pneumonia (MPP) and atelectasis. METHODS: A total of 75 children with MPP and atelectasis were divided into a good response group with 51 children and a poor response group with 24 children according to the clinical effect of BAL treatment. LASSO logistic regression analysis was used to investigate the factors influencing the clinical effect of BAL treatment. The receiver operating characteristic (ROC) curve and restricted cubic spline model analysis were used to evaluate the value of the course of the disease at the time of BAL treatment in predicting the clinical effect of BAL treatment. RESULTS: Compared with the good response group, the poor response group had a significantly lower percentage of lymphocytes in bronchoalveolar lavage fluid, a significantly higher proportion of children with atelectasis of two or more lung lobes or stenosis of the bronchial cavity or opening caused by inflammation, and a significantly longer course of the disease at the time of BAL treatment and azithromycin treatment (P<0.05). The LASSO logistic regression analysis showed that a prolonged course of the disease at the time of BAL treatment (OR=1.23), atelectasis of two or more lung lobes (OR=11.99), and stenosis of the bronchial cavity or opening caused by inflammation (OR=5.31) were independent risk factors for poor clinical effect of BAL treatment (P<0.05). The ROC curve analysis showed that the course of disease of ≥11.5 days at the time of BAL treatment suggested a poor clinical effect of BAL treatment, with a sensitivity of 91.7% and a specificity of 54.9%. The restricted cubic spline model analysis showed that there was a non-linear dose-response relationship between the course of disease at the time of BAL treatment and the clinical effect of BAL treatment (P<0.05). CONCLUSIONS: Early BAL treatment may have a good clinical effect in children with MPP and atelectasis. Atelectasis of two or more lung lobes and inflammation-induced stenosis of the bronchial cavity or opening shown under bronchoscope may indicate a poor clinical effect of BAL treatment.

[Value of lactate dehydrogenase in predicting refractory Mycoplasma pneumoniae pneumonia in children: an evaluation based on decision curve analysis and dose-response analysis].

OBJECTIVE: To study the value of lactate dehydrogenase (LDH) in predicting refractory Mycoplasma pneumoniae pneumonia (RMPP) in children. METHODS: Propensity score matching was used to select 73 children with RMPP (refractory group) and 146 children with non-refractory Mycoplasma pneumoniae pneumonia (common group). The logistic regression analysis, restricted cubic spline model, and decision curve analysis were used to analyze the clinical value of LDH in predicting RMPP. RESULTS: There were significant differences in the incidence of high fever, white blood cell count, platelet count, percentage of neutrophils, and serum levels of C-reactive protein, procalcitonin, hemoglobin, albumin, glutamic-pyruvic transaminase, aspartate aminotransferase and LDH (P<0.05). There were also significant differences between the two groups in the Mycoplasma pneumoniae-DNA load in nasopharyngeal aspirates and the incidences of pleural effusion, pulmonary consolidation, atelectasis, shortness of breath and skin lesions (P<0.05). The multivariate logistic regression analysis showed that high fever, hemoglobin level, LDH level, and pulmonary consolidation were independent predictive factors for RMPP (OR=10.097, 0.956, 1.006, and 3.756; P<0.05). The results of the restricted cubic spline analysis showed a non-linear dose-response relationship between the continuous changes of LDH and the development of RMPP (P<0.01). The decision curve analysis showed that LDH had an important clinical value in predicting RMPP. CONCLUSIONS: LDH is an independent predictive factor for the development of RMPP and its intensity of association with the development of RMPP exhibits a non-linear dose-response relationship.

[Clinical features of children with bronchial asthma complicated by pulmonary fungal infection and risk factors for pulmonary fungal infection].

OBJECTIVE: To study the clinical features of children with bronchial asthma complicated by pulmonary fungal infection and the risk factors for pulmonary fungal infection. METHODS: A retrospective analysis was performed for the clinical data of 150 children with bronchial asthma who were admitted from January 2015 to June 2018. Among these children, 75 had pulmonary fungal infection (fungal infection group) and 75 did not have such infection (control group). The distribution of pathogenic fungi, clinical symptoms/signs and treatment outcome were recorded for the fungal infection group. The multivariate logistic regression analysis was used to investigate the risk factors for pulmonary fungal infection. RESULTS: A total of 69 pathogenic fungi were detected in 75 children in the fungal infection group, among which Candida albicans had the highest detection rate of 61%. Major clinical symptoms were cough (93%), persistent high fever (56%), wheezing (49%) and dyspnea (48%). Major signs were dry and moist rales (43%) and moist rales (29%). Parts of children had hepatosplenomegaly. Among the 75 children in the fungal infection group, 39 were markedly improved, 26 were improved, 7 had no response, and 3 experienced aggravation and then died. Age <3 years, comorbidities of nasosinusitis and/or allergic rhinitis, asthma attacks of >3 times during hospitalization, intravenous administration of glucocorticoids, non-rational use of antibiotics, mechanical ventilation and prolonged hospital stay were independent risk factors for pulmonary fungal infection in children with asthma (OR=4.865, 3.241, 2.255, 3.725, 3.568, 1.549, 3.808; P<0.05). CONCLUSIONS: Pulmonary fungal infection should be considered for asthmatic children with cough, persistent high fever, obvious dry and moist rales and hepatosplenomegaly. The asthmatic children with an age of <3 years, comorbidities of nasosinusitis and/or allergic rhinitis, asthma attacks of >3 times during hospitalization, intravenous administration of glucocorticoids, non-rational use of antibiotics, mechanical ventilation or prolonged hospital stay have a higher risk for secondary pulmonary fungal infection.

[Clinical effect of fluticasone propionate, montelukast sodium and ketotifen in treatment of cough variant asthma in children].

OBJECTIVE: To study the clinical effect of different combinations of fluticasone propionate (Flu), montelukast sodium (Mon) and ketotifen (Ket) in the treatment of children with cough variant asthma (CVA). METHODS: A total of 280 children with CVA who were admitted to the department of respiratory medicine from June 2015 to January 2018 were randomly divided into Flu+Mon+Ket, Flu+Mon, Flu+Ket, Mon+Ket, Flu, Mon and Ket groups, with 40 children in each group. The children in each group were given corresponding drug(s), and the course of treatment was 3 months for all groups. The condition of cough, cough symptom score, pulmonary function and adverse drug reactions were evaluated after 2 and 3 months of treatment. The children were followed up to observe recurrence. RESULTS: After treatment, cough symptom score tended to decrease in all 7 groups, with increases in percentage of forced expiratory volume in 1 second (FEV1%) and percentage of predicted peak expiratory flow (PEF%). After 2 months of treatment, the Flu+Mon+Ket group had a significantly lower cough symptom score and significantly higher FEV1% and PEF% than the other groups (P<0.05). After 2 and 3 months of treatment, the Ket group had a significantly higher cough symptom score and significantly lower FEV1% and PEF% than the other groups (P<0.05). After 3 months of treatment, there were no significant differences in cough symptom score, FEV1% and PEF% among the other groups (P>0.05). There was a low incidence rate of adverse events in all 7 groups, and there was no significant difference among the 7 groups (P>0.05). The Ket group had a significantly higher recurrence rate of cough than the other groups (P<0.001), while there was no significant difference in this rate among the other groups (P>0.0024). CONCLUSIONS: For children with CVA, a combination of Flu, Mon and Ket has a better clinical effect than a combination of two drugs and a single drug at 2 months of treatment and is safe. After 3 months of treatment, Flu or Mon alone has a similar effect to drug combination. Ket alone has a poor clinical effect and a high recurrence rate after drug withdrawal.

SAMD9L inactivation promotes cell proliferation via facilitating G1-S transition in hepatitis B virus-associated hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a highly malignant cancer with poor prognosis, and driver genes harboring genetic lesions and/or expression dysregulation contribute to hepatocarcinogenesis. Sterile Alpha Motif Domain-containing 9-like (SAMD9L) was a novel identified mutated gene in our previous study on exome sequencing of hepatitis B virus (HBV)-associated HCC, but its expression and role in HCC remain unknown. Here, we demonstrated that SAMD9L was frequently inactivated by somatic mutations, and that its expression was deregulated in HCC patients with hepatitis B virus (HBV) infection. SAMD9L knockdown significantly promoted cell proliferation, colony formation of SK-hep-1, QGY-7701, BEL-7721 and MHCC-97H HCC cells. Furthermore, SK-hep-1 and MHCC-97H cells with stable SAMD9L knockdown exhibited enhanced tumorigenicity in athymic mice. Interestingly, SAMD9L silence facilitated G1-S transition of cell cycle progression and led to the elevated activity of Wnt/ß-catenin pathway. Collectively, these findings highlight a novel tumor-suppressive role of SAMD9L inactivation by somatic mutation and decreased expression in human HBV-related HCC.

RNA interference against cancer/testis genes identifies dual specificity phosphatase 21 as a potential therapeutic target in human hepatocellular carcinoma.

UNLABELLED: Cancer/testis (CT) antigens have been considered therapeutic targets for treating cancers. However, a central question is whether their expression contributes to tumorigenesis or if they are functionally irrelevant by-products derived from the process of cellular transformation. In any case, these CT antigens are essential for cancer cell survival and may serve as potential therapeutic targets. Recently, the cell-based RNA interference (RNAi) screen has proven to be a powerful approach for identifying potential therapeutic targets. In this study we sought to identify new CT antigens as potential therapeutic targets for human hepatocellular carcinoma (HCC), and 179 potential CT genes on the X chromosome were screened through a bioinformatics analysis of gene expression profiles. Then an RNAi screen against these potential CT genes identified nine that were required for sustaining the survival of Focus and PLC/PRF/5 cells. Among the nine genes, the physiologically testis-restricted dual specificity phosphatase 21 (DUSP21) encoding a dual specificity phosphatase was up-regulated in 39 (33%) of 118 human HCC specimens. Ectopic DUSP21 had no obvious impact on proliferation and colony formation in HCC cells. However, DUSP21 silencing significantly suppressed cell proliferation, colony formation, and in vivo tumorigenicity in HCC cells. The administration of adenovirus-mediated RNAi and an atelocollagen/siRNA mixture against endogenous DUSP21 significantly suppressed xenograft HCC tumors in mice. Further investigations showed that DUSP21 knockdown led to arrest of the cell cycle in G1 phase, cell senescence, and expression changes of some factors with functions in the cell cycle and/or senescence. Furthermore, the antiproliferative role of DUSP21 knockdown is through activation of p38 mitogen-activated protein kinase in HCC. CONCLUSION: DUSP21 plays an important role in sustaining HCC cell proliferation and may thus act as a potential therapeutic target in HCC treatment.

[Diagnostic values of bronchoscopy and multi-slice spiral CT for congenital dysplasia of the respiratory system in infants: a comparative study].

OBJECTIVE: To investigate and compare the diagnostic values of bronchoscopy and multi-slice spiral computed tomography (CT) for congenital dysplasia of the respiratory system in infants. METHODS: Analysis was performed on the clinical data, bronchoscopic findings and multi-slice spiral CT findings of 319 infants (≤1 years old) who underwent bronchoscopy and/or multi-slice spiral CT and were diagnosed with congenital dysplasia of the respiratory system. RESULTS: A total of 476 cases of congenital dysplasia of the respiratory system were found in the 319 infants, including primary dysplasia of the respiratory system (392 cases) and compressive dysplasia of the respiratory system (84 cases). Of the 392 cases of primary dysplasia of the respiratory system, 225 (57.4%) were diagnosed by bronchoscopy versus 167 (42.6%) by multi-slice spiral CT. There were significant differences in etiological diagnosis between bronchoscopy and multi-slice spiral CT in infants with congenital dysplasia of the respiratory system (P<0.05). All 76 cases of primary dysplasia of the respiratory system caused by tracheobronchomalacia were diagnosed by bronchoscopy and all 17 cases of primary dysplasia of the respiratory system caused by lung tissue dysplasia were diagnosed by multi-slice spiral CT. Of the 84 cases of compressive dysplasia of the respiratory system, 74 cases were diagnosed by multi-slice spiral CT and only 10 cases were diagnosed by bronchoscopy. CONCLUSIONS: Compared with multi-slice spiral CT, bronchoscopy can detect primary dysplasia of the respiratory system more directly. Bronchoscopy is valuable in the confirmed diagnosis of tracheobronchomalacia. Multi-slice spiral CT has a higher diagnostic value for lung tissue dysplasia than bronchoscopy.

[Effect of clinical pathway management on pediatric pneumonia].

OBJECTIVE: To investigate effect of clinical pathway management on pediatric pneumonia. METHOD: Data were colleted from children hospitalizated with bronchial pneumonia, bronchiolitis, mycoplasma pneumonia in Center of Respiratory Disorders in Children's Hospital of Chongqing Medical University from January 2011 to December 2012. According to implement of clinical pathway management, all patients were divided into pathway management group (n = 405) and non-pathway management group (n = 503). Length of stay, costs of hospitalization, clinical effect and use of antibiotics were compared in these two groups. RESULT: In pathway management group, average length of stay of children with bronchial pneumonia and bronchiolitis was (6.1 ± 1.6) d and (6.2 ± 1.5) d respectively. While in non-pathway management group, length of stay was (7.2 ± 1.9) d and (7.3 ± 1.5) d (P = 0.000). There was no significant difference in length of stay between these two groups of children with mycoplasma pneumonia [ (6.9 ± 1.8) d vs.(7.7 ± 2.5) d] (P = 0.198). Costs of auxiliary tests in pathway management group was slightly higher than that in non-pathway management group. While other costs in pathway management group were significantly lower than those in non-pathway management group. Total costs of hospitalization of patients with these three diseases in pathway management group and non-pathway management group were ¥(4609 ± 1225) vs ¥ (5629 ± 1813) , ¥ (5006 ± 1250) vs. ¥ (5686 ± 1337), ¥ (4946 ± 1259) vs. ¥ (6488 ± 3032) respectively. There was a significant difference (P < 0.05). Percentages of antibiotics use in two groups were 70.9% vs.99.4%, 45.7% vs.93.4% and 96.2% vs.100.0%. Antibiotics related indicators such as mean number of day of use, ratio of combination and grade of antibiotics were significantly higher in pathway management group compared to non-pathway management group (P < 0.01). There was no significant difference in other indicators like clinical effect and unscheduled readmission in 30 days between two groups (P > 0.05). CONCLUSION: Clinical pathway management can regulate medical behaviors through reduction of medical costs, avoidance of excessive laboratory tests and therapy, and regulation of antibiotic use.

[Causes of stopping subcutaneous specific immunotherapy in asthmatic children].

OBJECTIVE: To improve the compliance with subcutaneous specific immunotherapy (SCIT) by analyzing the causes of stopping SCIT in asthmatic children. METHODS: A telephone follow-up was conducted in the asthmatic children who received SCIT but did not finished the 3-year course of treatment from June 2005 to October 2010, so as to analyze the causes of stopping SCIT. RESULTS: A total of 616 asthmatic children received SCIT, and 322 (52.2%) of them stopped SCIT.A total of 127 cases (39.4%) of the 322 children received telephone follow-up. In the 127 children, 53 (41.8%) stopped the SCIT for the reason of bad effecacy, 29 (22.8%) for remission of asthma,12 (9.4%) for expensive fees, 10 (7.9%) for complex process of treatment, 10 (7.9%) for adverse reaction, 9 (7.1%) for long distance from the hospital, and 4 (3.1%) for having no time for treatment. And 69 (54.3%) of them stopped SCIT in the first year, 28 (22.1%) in the second year, and 30 (23.6%) in the third year. Currently, 85 cases (66.9%) of the 127 asthmatic children were up to the control level, and the other 42 cases were not. There was significant difference in the control level of asthma berween the group receiving treatment with regular inhaled corticosteroids (ICS) and the group receiving treatment with irregular ICS (P<0.01). CONCLUSIONS: Bad efficacy, remission of asthma, expensive fees, complex process of treatment, and adverse reaction are the main reasons contributing to the stop of SCIT in asthmatic children. To improve the compliance with SCIT, It is important to make the patients and their parents understand the long treatment course and slow effect of SCIT, encourage them to use objective indices for evaluating the state of asthma, and effectively prevent and treat the adverse reactions.

Exome sequencing of hepatitis B virus-associated hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and shows a propensity to metastasize and infiltrate adjacent and more distant tissues. HCC is associated with multiple risk factors, including hepatitis B virus (HBV) infection, which is especially prevalent in China. Here, we used exome sequencing to identify somatic mutations in ten HBV-positive individuals with HCC with portal vein tumor thromboses (PVTTs), intrahepatic metastases. Both C:G>A:T and T:A>A:T transversions were frequently found among the 331 non-silent mutations. Notably, ARID1A, which encodes a component of the SWI/SNF chromatin remodeling complex, was mutated in 14 of 110 (13%) HBV-associated HCC specimens. We used RNA interference to assess the roles of 91 of the confirmed mutated genes in cellular survival. The results suggest that seven of these genes, including VCAM1 and CDK14, may confer growth and infiltration capacity to HCC cells. This study provides a view of the landscape of somatic mutations that may be implicated in advanced HCC.

[Construction and identification of yeast expression plasmids containing mouse signal transducers and activators of transcription-4/6].

AIM: To construct yeast expression plasmids containing mouse STAT4/6 gene for further study of their interaction with other proteins in yeast two-hybrid system. METHODS: Mouse STAT4/6 genes were amplified by PCR and T-A was cloned with pMD19-T simple vector and then was cloned into yeast expression vector pGADT7 cut with incision enzymes and treated with CIAP. The yeast expression vector pGADT7 was identified by enzyme cutting and sequencing. The yeast expression plasmids pGADT7-STAT4/6 were transformed into AH109 yeast cells and the expression of the STAT4/6 fusion proteins was detected by Western blot. Their toxicity and self-activation were also detected. RESULTS: Mouse STAT4/6 genes were successfully amplified and cloned into pMD19-T simple vector and pGADT7. Sequencing analysis revealed that both plasmids met the design of the study. The yeast expression plasmids pGADT7-STAT4/6 were successfully transformed into AH109 yeast cells, without toxicity or self-activation. The expression of STAT4/6 fusion proteins was confirmed by Western blot. CONCLUSION: The yeast expression plasmids pGADT7-STAT4/6 are successfully constructed and can be applied in the detection of their interaction with other proteins.

[Biological effects of lipopolysaccharide, transforming growth factor-beta1 on murine bone marrow-derived dendritic cells].

AIM: To explore method of stimulating murine bone marrow-derived dendritic cells by lipopolysaccharide (LPS), transforming growth factor-beta1 (TGF-beta1) and to study their biological character. METHODS: Murine bone marrow-derived dendritic cells were cultivated with cytokine GM-CSF and IL-4 for 6 days, BMDC was stimulated by control, LPS, TGF-beta1, LPS +TGF-beta1 for 48 hours respectively.Morphological characters of BMDC were observed by a inversed microscope, surface molecules such as CD(11C), CD(80), CD(86) and MHC II were detected by flowcytometry, Interleukin-6 and interleukin-12 p70 in co-culture medium was quantified by ELISA. RESULTS: In LPS group it presented the most typical DC morphology with the highest expression of CD(80), CD(86) and MHC II, the strongest ability in mixed lymphocyte reaction, higher level of IL-6 and IL-12 p70 compared with control, TGF-beta1, LPS+TGF-beta1 (P<0.05). While in TGF-beta1 group it presented the less typical DC morphology with the lower expression of CD(80), CD(86), MHC II, weaker ability in mixed lymphocyte reaction, and lower levels of IL-6 and IL-12 p70 compared with control and LPS (P<0.05). CONCLUSION: LPS can stimulate maturation of BMDC in its late differentiation which makes it presents a more significant biological characteristics.TGF-beta1 can inhibit maturation but not differentiation of BMDC thereby can prevent its biological characteristic presentation.

[Treatment of zygomatio- orbital fractures with small local incisions: report of 32 cases].

PURPOSE: To evaluate the surgical result of small local incisions in the treatment of zygomatio-orbital fractures. METHODS: 32 patients with zygomatio-orbital fractures were operated on with two or more kinds of incisions, including subciliary incision, lateral eyebrow incision, transverse incision along the superior margin of zygomatic arch, oral vestibular incision, maxillary sinus incision, the original wound according to the area and type of the fractures. The fracture line was exposed via the small local incisions and fixed with micro-titanium plates. The therapeutic effectiveness was evaluated 3 to 6 months after operation. RESULTS: All patients had primary healing without apparent scars in the face,except that one patient developed mild ectropion and two had temporary paralysis of the temporal branches of the facial nerve. The function of the eyes recovered completely, while the jaw movement improved significantly. The facial appearance recovered to normal height and width and the bilateral face became symmetrical. CONCLUSION: Treatment of zygomatic-orbital fractures with small local incisions is minimally invasive and the facial appearance and function can be recovered significantly with few complications.