To detect the levels of miR-146a and miR-155 in different samples from chronic hepatitis B (CHB), reveal whether there is a correlation between the 2 miRNAs in different samples, and to provide a theoretical basis for sample choice of miRNA research in liver.
 Methods: Real-time PCR was conducted to examine the expression of miR-146a and miR-155 in the plasma, peripheral blood mononuclear cell (PBMC), and liver tissues from 41 CHB patients who underwent nucleoside analogues antiviral therapy for 104 weeks. Correlations between the levels of miR-146a and miR-155 among the 3 samples were analyzed.
 Results: The expressions of miR-146a and miR-155 in the plasma, PBMC and liver tissues were significantly down-regulated at the 104th week than those at the baseline (all P<0.05). There was a correlation in the expression of miR-146a between plasma and liver tissues (r=0.560, P=0.007), PBMC and liver tissues (r=0.428, P=0.047) at baseline. There was a correlation in the expression of miR-155 between plasma and liver tissue (r=0.587, P=0.004), PBMC and liver tissue (r=0.483, P=0.023) at baseline. The expressions of miR-146a and miR-155 between the plasma and PBMC were not correlated (P>0.05).
 Conclusion: Compared with PBMC, miR-146a and miR-155 from plasma can better reflect the expression in the liver tissues, suggesting that plasma can be applied in the mechanism research on miR-146a and miR-155 in the liver diseases instead of liver tissues.
Hepatitis B, Chronic ; genetics ; Humans ; Leukocytes, Mononuclear ; MicroRNAs ; genetics ; Real-Time Polymerase Chain Reaction

Objective:To investigate expression profiles of the plasma exosomal miRNAs of the chronic hepatitis B (CHB) patients with persistently normal alamine aminotransferase (PNALT) for the first time and try to find exosomal miRNAs which could reflect liver inflammation better.Methods:Five CHB patients with liver tissue inflammation grade ≥A2 of PNALT and 5 CHB patients with liver tissue inflammation grade ＜A2 of PNALT were enrolled and their blood samples were collected.The exosomes were extracted from these blood samples and measured by electron microscope to determine the extraction effect.The exosomal miRNAs were extracted and sent for high throughput sequencing,and the expression of exosomal miRNAs in the 2 groups of patients was analyzed.Results:Under the electron microscope,exosomes were small membranous vesicles with 30-100 nm in diameter.The peak value of particle size ranged from 10 to 100 nm.High throughput sequencing showed that there were 591 differentially expressed exosomal miRNAs between the 2 groups.Compared with the control group,18 exosomal miRNAs were up-regulated and 6 exosomal miRNAs were down-regulated in PNALT patients with the liver tissue inflammation grade ≥ A2.Conclusion:Exosomal miRNAs in the CHB patients with PNALT who have the different grades of liver inflammation are differently expressed.Some of the differently expressed exosomal miRNAs are expected to be sensitive biomarkers for timely assessment of liver inflammation in the CHB patients with PNALT.

Objective To evaluate the accuracy and feasibility of time-resolved immunofluorometric assay (TRI FA) for detection of HBsAg based on Abbott automated chemiluminescence immunoassay(CMIA),so as to carry out this project in primary hospitals,and provide reference for individual antiviral strategy and prediction of therapeutic effect.Methods Serum of 157 patients infected with hepatitis B virus were detected with CMIA and TRIFA,specimens with HBsAg titers exceeding the detection limit were firstly diluted,then performed quantitative analysis.HBsAg levels were divided into 4 groups:≤100 IU/mL,101-1 000 IU/mL,1 001-20 000 IU/mL,and ＞ 20 000 IU/mL,quantitative correlation between two methods was analyzed.Results The linear regression equation of two methods was Y=2.323X-896.3,correlation coefficent r=0.943,P＜0.001.CMIA was as a reference,4 groups were divided for analysis,results showed that when detected specimens was at low concentration of HBsAg,TRIFA value was low compared with CMIA method,while detected specimens was at high concentration of HB sAg,CMIA value was high,two reagents had good consistency in the detection of different concentrations of HBsAg(both P＜0.05),when concentration was at 1 001-20 000 IU/mL,consistency was the best.Conclusion The accuracy of two reagents in the quantitative detection of HBsAg is similar,and the best correlation of detection value is 1 000-20 000 IU/mL.TRIFA assay has wide application for its low-cost and easy to be operated,which is especially suitable for primary hospitals.

To analyze influenza pathogen spectrum in Yunnan province during 2009-2014 years, and analyze HA and NA genes of influenza A H1N1. Analysis was made on the monitoring date of influenza cases in Yunnan province in recent 6 years, 23 strains of influenza virus of HA and NA gene was sequenced and analyzed by MEGA 5 software to construct phylogenetic tree. 4 times of influenza AH1N1 epidemic peak were monitored from 2009-2014 years in Yunnan Province, as the nucleic acid detection results of influenza A H1N1 accounted for 28.8% of the total. The sequencing result showed that HA and NA gene were divided into 3 groups, one was detected with H275Y mutation strains. Influenza A H1N1 is one of the important subtypes in Yunnan province and their genes have divided into three branches during the period of 2009-2014 years, the vast majority of influenza a H1N1 are still sensitive to neuraminidase inhibitors.
China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Mutation ; Neuraminidase ; genetics ; metabolism ; Phylogeny ; Viral Proteins ; genetics ; metabolism

Purpose To investigate the clinicopathologic features of cervical adenoid basal cell carcinoma. Methods Clinical and pathological data were analyzed in 12 cases of cervical adenoid basal cell carcinoma and the relevant literatures were reviewed. Results The age of 12 cases of with cervical adenoid basal cell cancer patients ranged from 36~70 years ( mean:53. 3 years) , with a medi-an age of 51. 5 years. Amongst 12 patients who contained no gross definite lesion, 5 patients (41. 7%) had the symptoms of vaginal bleeding. 8 cases of patients underwent hysterectomy, while the other four received cervical conization. Cervical intraepithelial neopla-sia ( CIN) lesions were observed in all 12 cases. The tumor cells were small and uniform, with dark oval nuclei without conspicuous nucleoli and scanty cytoplasm. Tumor infiltrated into the stroma in nests and cords. Glandular differentiation within tumor nests were seen in 12 cases, including two cases of squamous cell differentiation accompanied by partial and minor interstitial edema. Peripheral palisading cells around tumor nests were also found. The infiltrating depth of tumor ranged from 0. 5~10 mm with the average 4. 12 mm. Cancer embolis were available in 2 cases with no lymph node metastasis. 10 cases followed up for 3~78 months with no recur-rence and metastasis, but 2 cases were lost. Conclusion Adenoid basal cell carcinoma is a rare uterine cervical tumor found in post-menopausal women, which are often accompanied with CIN. This tumor has a favorable prognosis and should be clearly separated from adenoid cystic carcinoma and other tumors which also have infiltrative growth pattern.

Objective To observe and evaluate the effects of the deep hypothermic circulatory arrest(DHCA) and regional cerebral perfusion(RCP) in pediatric aortic arch surgery.Methods According to different methods of CPB,70 infants less than 3-month-old with CoA or IAA were undergone corrective surgery with DHCA or RCP.The bypass time,aortic clamp time,DHCA or RCP time,ventilation time,ICU stay time and post-operative complications were recorded and compared between two groups.Results The incidence of neurological complications was significantly higher in DHCA group.The CPB time was significantly longer in the RCP group,and the RCP time was significantly longer than DHCA time.Blocking time,ventilator intubation time,ICU residence time,postoperative renal dysfunction,low cardiac output,puhnonary inflammation and hospital mortality was no significant difference between the two groups.Conclusion RCP is an effective cerebral protection technique.Compared with DHCA,RCP works better in sustained brain cerebral perfusion and is suitable for complex aortic arch operation in children.It has a better effort in protection of the neurological system than DHCA.

Objective To understand the viral etiology of acute respiratory infection in Kunming area. Methods We collected the nasopharyngeal swab of patients with acute respiratory tract infection,and used multiple reverse transcription-polymerase chain reaction (RT-PCR) method to detect 15 kinds of respiratory viral pathogens. Results Among the 600 samples,144 strains of viruses were detected, the positive rate was 24%,among which the highest positive rate was RSV (49/600,8.2%),followed by PIV (32/600,5.3%) HRV (27/600,4.5%) and IFV27 (27/600,4.5%) . The respiratory virus infection situation was different in every age group, groups of the highest virus positive rate was ≤1 age group (72/216, 33.3%);The respiratory virus infection situation in different seasons was different, the virus positive rate of the first quarter was the highest (85/144, 59%) . Conclusion RSV was the main virus pathogen of acute respiratory tract infections in Kunming area in 2011 years, the detection rate in sick children was the highest among all patients;the detection rate in the first quarter was higher than other quarters.

Objective The use of extracorporeal membrane oxygenation (ECMO) as a treatment for the failure of cardiopulmonary function after cardiac surgery is increasing and has been reported to be 3% to 5% in the cases with congenital heart disease. We reviewed our experience with ECMO in children who received heart surgery for congenital heart disease and complicated with severe heart failure postoperatively. Methods Eight patients received ECMO, seven was due to the failure to wean from bypass and one had fulminant myocarditis. Import membrane oxygenator,veno-arterial mode ECMO and right atriumascending aortic cannulation were used in 7 cases and peripheral cannulation via femoral veno-artery route was used in 1 case.Supportive intervention persisted from 65 to 498 hours, with flow rate maintained at 80 to 120 ml per minute per kilogram body weight. Results Five patients died, with a mortality of 62.5%, and 3 cases discharged, with a survival rate of 38%. Bleeding occurred in 5 cases, thrombosis occurred in 2 cases, hemolysis was identified in 1 case and DIC was observed in 1 case.One case had liver failure and 2 cases had malnutrition. Oxygenator plasma leakage occurred in 2 cases. Mean arterial blood pressure increased significantly after the establishment of ECMO as compared with that before the procedure [( 60.2 ± 7.8 )mmHg vs. (48. 1 ± 5.2 ) mmHg, P≤0.05]. The arterial concentration of lactate decreased significantly, from (5. 1 ± 0. 8 )mmol per liter before ECMO to ( 3.6 ±0. 5 )mmol per liter after ECMO, P ＜0.05. Conclusion For patients who survived the congenital heart surgery and no residual anatomic deformity, ECMO can be used as early as possible as a treatment for severe heart failure which resulted from coexistent of left and right ventricular and pulmonary insufficiency. An overall mortality may be decreased by ECMO technique as it plays a substitution role for gas exchange in the lung. As a result, the concentration of oxygen and the airway pressure used during ventilation, and the resultant lung injury can be reduced. Appropriate strategies involve transfusion of fresh platelet and packed red blood cells, replacement of frozen plasma and blood products, as well as rational use of vasoactive drugs and heparin, and maintaining a stable internal environment. Following strategies are also recommended: using continuous arterio-venous hemofiltration and durable heparin-coated membrne oxygenator, reducing hemorrhagic complications, monitoring pressure on both side of the film, identifying plasma leakage carefully and reducing the mechanical complications.

Objective To investigate the effect of TGFβ1 siRNA on hepatic satellite cells (HSCs) activation, proliferation and extracellular matrix production. Methods The TGFβ1, siRNA plasmid was transfected into HSCs with Lipofectamine 2000. The supernatant and HSCs were collected after incubation for 72h. The expression of TGFβ1, and a-SMA protein in HSCs was detected by. Western blotting. The expression of type Ⅰ and Ⅲ collagen mRNA was detected by RT-PCR. The cell proliferation was assayed by MTT method. Contents of typeⅣ collagen and hyaluronic acid in supernatant were determined by radioimmuno-assay.Results Compared with scrambled control group, the TGFβ1, and a-SMA protein expression,the activity of HSCs proliferation,the expression of typeⅠ and Ⅲ collagen mRNA,and the contents of type Ⅳ collagen and hyaluronic acid in supernatant were reduced in TGFβ1, siRNA group by (79±5)%,(55±4)%, (25±4)% ,(63±6)% ,(57±4)% ,(53±8)% ,(46±8)% ( P<0.01),respectively.Conclusion TGFβ1, siRNA could significantly reduce the expression of TGFβ1,inhibited HSC activation,proliferation and extracellular matrix production.

BACKGROUND: Transplantation of microencapsulated rabbit Schwann cells in the rat spinal cord can relieve inflammatory reaction, promote spinal cord regeneration, but the precise mechanisms remain unclear. OBJECTIVE:To observe basic fibroblast growth factor (bFGF)expression and movements recovery following transplantation of microencapsulated rabbit Schwann cells in rat spinal cord. METHODS: The sciatic nerves taken out from rabbits wore digested with mixed enzyme and were made into Schwann cells suspension. Then we used air-jet method to make Schwann cells microcapsule. Using the same method, empty microcapsule was made. Sprague Dawiey rats were randomly divided into cell group, empty microcapsule group and microcapsule group. Conducted by hemisection injury of spinal cord,the rats in cell group,empty microcapsule group and microcapsule group were implanted with gelatin sponge with 10μL Schwann cells suspension, gelatin sponge with 10 μL empty microcapsule and 10 μL microencapsulated Schwann cells. Normal group was left intact. After operation, we observed hindlimb movements recovery in rats with the Basso, Beattie, and Bresnahan (BBB) scale. Meanwhile,a set of sections were stained immunohistochemically for bFGF expression, another set of sections wore stained for hematoxylin-eosin and Nissal. RESULTS AND CONCLUSION: After spinal cord injury, rat right hindlimb affected paralysis immediately. At 7, 14 and 28 daysfollowing transplantation,motor function in rat hindlimb was significantly recovered, and the BBB scores were significantly higher in microencapsulated schwenn cells than in cell and empty microcapsule group (P < 0.05 or P < 0.01). bFGF positive products were mainly distributed in cytoplasm of the spinal neuron and nucleus of neuroglical cell. The numbers of bFGF positive glial cells mainly appeared surrounding the spinal cord injured site on days 1, 3, 7 and rose to its peak on day 3 and began to appear in neuronal calls on day 14. The number of bFGF positiv cells in microcapsule group was significantly superior to that in cell group and empty microcapsule group. From then on, the bFGF expreSsion was significantly decreased in each group. These indicated that transplantation of microencapsulated Schwann cells can inhibit the immunological rejection after xenotransplantation, suppress inflammatory reaction, improve the expression of bFGF, increase hindlimb movements recovery and spinal cord regeneration after spinal cord injury.