[ ABSTRACT] AIM:To investigate the biological characteristics of newborn rabbit tracheal chondrocytes in vitro. METHODS:Newborn rabbit tracheal chondrocytes were obtained by the method of enzyme digestion, and then cultured in monolayer in vitro.Morphological and growth observations were performed under inverted phase contrast microscope.The ultrastructures of the cells were observed under scanning electron microscope and transmission electron microscope.The bi-ological characteristics of secreted extracellular matrix components were detected by real-time PCR, immunocytochemistry staining and toluidine blue staining.RESULTS: Newborn rabbit tracheal chondrocytes isolated and cultured in vitro showed short triangular or irregular shapes, and adherent growth very well.The ultrastructures of the cells showed pore and abundant cytoplasm and organelles, with a lot of protein secretions in the cells.The chondrocytes expressed the mRNA of collagen I, collagen II and proteoglycans, mainly collagen II and proteoglycans.Immunocytochemistry staining showed col-lagen II and SOX9 positive, and collagen I weakly positive.Toluidine blue staining was also positive.CONCLUSION:Enzyme digestion and monolayer culture are suitable method to obtain newborn rabbit tracheal chondrocytes.These cells, secreting extracellular matrix components, are able to be selected as seed cells for tissue engineering of trachea in vitro, and used to study the therapeutic method for neonatal rabbit tracheal stenosis.

Objective To evaluate the role of transesophageal echocardiography (TEE) in perioperative period of pediatric patients with congenital heart disease (CHD).Methods From July 2011 to December 2012,TEE was used in 393 pediatric patients(≤ 14 years) with congenital heart disease in perioperative period.We make a retrospective review with the clinical data of these cases.Results Operative schemes or therapeutic schedules of 60 patients(15.3％) were altered according to TEE.By preoperative TEE,the diagnosis of transthoracic echocardiography (TTE) of 4 cases(1.0％) were amended,4 cases (1.0％) were complemented,and 4 cases (1.0％) got the auxiliary diagnosis,among which operative schemes of 11 cases (2.8％)were altered.During the operate,29 cases(7.4％) found residual problems,11 cases(2.8％) got the auxiliary diagnosis.By postoperative TEE,7 cases(1.8％) found residual problems,1 case(0.3％) got the auxiliary diagnosis.Complications occurred in 9 cases(2.3％) of the 393 patients.Oral and pharyngeal mucous membrane bleeding occurred in 7 cases (1.8％),inadvertent tracheal extubation in 2 cases(0.5％).Conclusion TEE plays an important role in confirming preoperative diagnoses,formulating surgical plans,evaluating immediate operative results,identifying patients with residual defects and guiding the therapeutic schedule in perioperative period of pediatric patients with congenital heart disease.

Aim The effects of midazolam on the whole-cell sodium currents in rat sympathetic neurons were studied to explore the mechanisms where by midazolam mediates hypotension. Methods Whole-cell patch-clamp recordings were performed on enzymatically isolated rat superior cervical sympathetic neurons. Results Midazolam dose-dependently blocked the whole-cell sodium currents evoked by a voltage step to 0 mV from a holding potential of -80 mV with a mean drug concentration required to produce 50% current inhibition (IC50) values of 18.35 ?mol?L-1; Clinically relevant concentration of midazolam(0.3 ?mol?L-1) reduced sodium peak currents by 19.98%(P

Objective:The purpose of this study is to evaluate the frequency of MDSCs in peripheral blood of hepatocellular carcinoma patients and to investigate the clinical significance of change of MDSCs in the peripheral blood and provide new ways for e-valuating immune state and prognosis of hepatocellular carcinoma patients.Methods: Blood samples were obtained from 62 patients with HCC and 20 healthy donors.The phenotype of CD3,CD4,CD33,HLA-DR and Th1,Th2 immune subsets in peripheral blood of each group were observed by FCM methods.Results:There were statically different frequencies in the peripheral blood between hepato-cellular carcinoma and healthy control group,which the proportion of total CD3+T lymphocytes and CD3+CD4+T cells were lower and the proportion of CD33+HLA-DR-MDSCs was higher in hepatocellular carcinoma patients.( P<0.05 ).The increase of percentage of MDSCs was greater in patients at Stage C and D than in patients at stage A and B.Conclusion:The Th1/Th2 ratio in the PBMC were of imbalance and MDSCs was significantly increased in peripheral blood of hepatocellular carcinoma patients.The increase of MDSCs was significantly correlated with clinical stage.CD33+HLA-DR-MDSCs may play an important role in prediction in prognosis and tumor immune status of hepatocellular carcinoma.

Objective To evaluate the value of iterative modal reconstruction (IMR) for reducing radiation dose and controlling image quality in cardiac CT. Methods Ten pigs were included. All pigs were scanned on a 256?slice prospectively ECG?gated cardiac CT utilizing routine dose (group A) and tube current reduced by 30%(group B), 50%(group C) and 70%(group D), respectively. Filtered back projection (FBP), hybrid iterative reconstruction (iDose4) and IMR were used for all data, respectively. Image noise and contrast?to?noise ratio (CNR) of ascending aortic root were measured, while overall image quality and coronary artery image quality was rated (five point scale). All results reconstructed by FBP, iDose 4 and IMR were compared. Objective measurements were compared with one?way analysis of variance, and subjective assessments were compared with Kruskal?Wallis H test andχ2 test. Results Compared with that of FBP and iDose4, image noise of IMR was(15.1 ± 6.1),(18.8 ± 5.5),(22.1 ± 4.8)and(33.0 ± 4.0)HU, respectively in group A, B, C and D with significant reduction (F=82.77, 90.71, 96.59, 95.51 respectively, all P<0.01). Using IMR, groups A, B, C, D had higher CNR (42.0±11.1, 37.2±10.4, 31.4±8.7, 23.7±7.0;F=50.65, 53.55, 76.60, 57.36, all P<0.01) and overall image quality (5.0 ± 0.0, 4.8 ± 0.4, 4.6 ± 0.5, 4.5 ± 0.5;H=20.96, 15.63, 18.66, 23.56, all P<0.01) than FBP and iDose4. Using IMR, group A (100%, 40/40) and group B (100%, 40/40) had no significant difference (P>0.05) in the diagnosis rates of proximal coronary arteries compared with that using FBP and iDose4, while group C (100%, 40/40) and group D(92%, 37/40) had significantly increased diagnosis rates (χ2=20.05, 45.72, both P<0.01). The diagnosis rates of distal coronary arteries of IMR reconstruction which were 100%(50/50), 98%(49/50), 90%(45/50), 78%(39/50), respectively in groups A, B, C, D had significant increase compared with that of FBP and iDose4 reconstruction (χ2=7.39, 16.75, 34.62, 81.33, all P<0.05). Conclusions IMR can significantly reduce image noise, improve CNR and image quality compared with iDose4. Application of IMR can reduce radiation dose but without compromising image quality.

BACKGROUND: Bone marrow mesenchymal stem cells are important seeded cells for construction of tissue-engineered trachea, but there is no special surface marker. Therefore, identification of bone marrow mesenchymal stem cells is mostly based on morphology, phenotype antigen and the function of differentiation. OBJECTIVE: To explore the feasibility of the tracheal chondrogenic differentiation of bone marrow mesenchymal stem cells under a special condition through isolation, cultivation and identification of bone marrow mesenchymal stem cells. METHODS: Rabbit bone marrow was acquired in the sterile environment to isolate and culture bone marrow mesenchymal stem cells to passage 2 by bone marrow adherence and screening method. Flow cytometry identified the phenotype CD44, CD45 of bone marrow mesenchymal stem cells at passages 1 and 2. Rabbit tracheal samples were acquired in the sterile environment, the tracheal chondrocytes were isolated and cultured by enzyme digestion, and toluidine blue staining was used to detect aggrecan. Bone marrow mesenchymal stem cells were co-cultured with tracheal chondrocytes by Transwel and transforming growth factor β1. Cel morphology was detected under an inverted microscope. Real-time quantitative PCR and toluidine blue staining detected the extracel ular matrix components, such as type Ⅱ col agen and aggrecan.RESULTS AND CONCLUSION: After isolation and culture, cells were spindle and irregular in morphology, and passaged cells thrived that were gathered into a fish-like colony growth. For passage 1 bone marrow mesenchymal stem cells, the positive rates of phenotype antigen CD44 and CD45 were respectively 96.97% and 13.72%; for passage 2 cells, the positive rates of phenotype antigen CD44 and CD45 were 99.11% and 8.54%, respectively. Tracheal chondrocytes were positive for toluidine blue staining. The morphology of induced bone marrow mesenchymal stem cells changed from long fusiform to triangular or irregular shape, indicating the chondrocytes expressed type Ⅱ col agen and aggrecan, and toluidine blue staining was positive. These results showed bone marrow adherence and screening method could acquire bone marrow mesenchymal stem cells, and the purity of passage 2 cells is higher. Under a special condition, bone marrow mesenchymal stem cells have the potential of chondrogenic differentiation, and can be selected as seed cells for construction of tissue-engineered trachea.

Objective To investigate the effect of the nuclear factor (NF)-κBp65 antisense oligonucleotide (ASODN) on NF-κB activity and expression of interleukin(IL)-6 in hepatic stellate cells (HSC). Methods The HSC were separated from rats and cultured. The toxicity of NF-κBp65 ASODN on HSC were detected by Trypan blue exclusion staining and the NF-κB activity was determined by EMSA. The expressions of IL-6 mRNA and protein were meaured by RT-PCR and ELISA, respectively. Results In vitro, no toxicity of ASODN on HSC was observed at the concentrations of 0. 001 to 1.0 μmol/L. NF-κB activity was increased after stimulating HSC with tumor necrosis factor (TNF)α, whereas it was weakened in a dose dependent manner when HSC were cultured with ASODN (concentration from 0. 001 to 1.0 μmol/L). At the same time, the expressions of IL-6 mRNA and protein induced by TNFα were decreased after transfected with ASODN at concentrations of 0.001- 1. 0 μmol/L in a dose dependent manner. Conclusion ASODN may specifically inhibit either the activiy of NF-κB or expression of IL-6, which provides the theoretical basis that ASODN may use to treat fibrosis of the liver.

Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P

Objective To investigate the correlation between the indications,findings,interventions of fibrobronchoscopy(FB) in neonates and their correlative diseases with neonatal FB results and clinical data.Methods Retrospective case series of 243 consecutive patients of 28 days old or younger were investigated underwent FB for the first time from January 2010 to December 2014,at a tertiary care hospital.The common indications for FB and detection rate of respiratory tract diseases were collected.If the findings of FB had significant associations with premature birth and other diseases were analyzed.Associations between interventions and basic illnesses were also analyzed.Results Of the 243 patients undergoing 275 procedures of FB,201 cases were boys(73.1％).The age of FB was (13.34 ± 9.76) days and the weight was (3.08 ± 0.68) kg.Forty-five cases were premature infants (16.4％).A total of 254 procedures were found to have congenital diseases (92.4％),and 177 cases of them had congenital heart diseases (CHD) (64.4％).Common indications for FB were dyspnea(140 cases,50.9％),tachypnea(82 cases,29.8％),and stridor(71 cases,25.8％).A total of 188 upper airway lesions were found and the most common findings were laryngomalacia(56 cases,20.4％) and vocal cord paralysis(bilateral/unilateral,50 cases,18.2％).A total of 315 lower airway lesions were found and the most common findings were airway mucosal inflammation (98 cases,35.6％),trachea and main bronchial stenosis (73 cases,26.5 ％).A total of 21 cases (7.6％) underwent supraglottoplasty during or after FB,while 17 cases (6.2％) underwent tracheal dilation and 10 cases (3.6％) underwent tracheotomy.Compared with non-CHD neonates,neonates with CHD were statistically significantly less likely to have congenital lesions statistically,such as laryngomalacia(15.8％ vs.28.6％,P =0.012),bilateral vocal cord lesions(6.2％ vs.21.4％,P =0.000) and congenital laryngeal dysplasia(0 vs.7.1％,P =0.001).The tracheotomy(0 vs.10.2％,P =0.000) and supraglottoplasty(2.3％ vs.17.3％,P =0.000) were more rare.Nevertheless,they were more likely to have secondary lesions such as the left main bronchial stenosis caused by extrinsic compression (23.7％ vs.1.0％,P =0.000),abnormal bronchial anatomy(9.6％ vs.2.0％,P =0.018),left vocal cord paralysis(9.0％ vs.1.0％,P =0.008) and airway mucosal inflammation(41.8％ vs.24.5％,P =0.004).The tracheostenosis and main broncial stenosis (37.3％ vs.7.1％,P =0.000) with long-term intubation(78.5％ vs.58.2％,P =0.000) were more common.There was no significant difference between term neonates and premature infants in the detection rate of respiratory tract diseases (P ＞ 0.05),tracheotomy (0 vs.4.3 ％,P =0.322),supraglottoplasty (13.3 ％ vs.6.5 ％,P =0.205) or long-term i ntubation (80.0％ vs.69.6％,P =0.157).Complications caused by procedure were rare and mild.Conclusions FB can detect whether the neonates with dyspnea,tachypnea and stridor have laryngomalacia,vocal cord paralysis,airway mucous edema,tracheal and main bronchial stenosis and other signs,and FB may play an important role in diagnosis,treatment and prognosis evaluation of neonatal respiratory diseases.

Objective To summarize the clinical experience of surgical intervention for cardiac neoplasm in a fetus . Methods A 32-year-old pregnant woman was admitted to our hospital for complaint of fetal cardiac neoplasm .A separated het-erogenic cardiac occupying lesion was identigied at right atrium of the fetus by echocardiography , whose size is 2.85 cm ×2.25 cm, but the pathogenic origin still remained uncertain, maybe originate from ether pericardium or atrium.The annulus of tri-cuspid valve was compressed nearly 50% with the presence of amount of pericardial effusion.The fetal heart rate decreased at some fetal position resulting in the compression to the heart.So an Ex-utero Intrapartum Therapy(EXIT) procedure was per-formed under the supply of placenta at the 32 weeks of pregnancy.Cesarean section was performed with intact umbilicus and fe-tal circulation by obstetricians.Consequently, the median sternotomy of this fetus and pericardiotomy were performed , with 30 ml clear pericardial effusion drained .The tumor was confirmed to be giant right atrial neoplasm after the intraoperative explora-tion.Considering on the high risk of the cardiopulmonary bypass and limited time for EXIT , the giant atrial neoplasm was left alone with delayed sternum closure after the effectively decompression of the heart .The omphalotomy was successfully per-formed after the EXIT surgery.The neoplasm resection and the repair for its defect on right atrium were performed with cardiop-ulmonary bypass 2 days later.Results Convalesce of this mother was quite good after cesarean resetion .Hemodynamics of the premature baby was satisfatory after the resection of right atrial neoplasm which pathological report was benign hemangioma . Conclusion Via multiple disciplines collaboration , EXIT intervention for fetus is feasible and safe under adequate prepara-tion.