Volume 94  ·   Issue 42  ·  2014

human leucocyte antigen G (HLA-G) is a non-classical major histocompatibility complex (MHC-I). In recent years, the research on HLA-G genes and molecules is increasing day by day. Since 1989, Geraghty first reported successful cloning of HLA-G^[1]In 1998, the 1st International Conference on HLA-G was held at the Institute of Immunology of St. Louis Hospital in France. At that time, there were 208 HLA-G related articles, and now there are 1 738 articles. In China, many scholars have also carried out research and reports on HLA-G.

To explore the protective effects and mechanism of exosomes derived from bone marrow mesenchymal stem cells (BM-MSC) on ischemia reperfusion injury (IRI) in rats.

Rat-MSC were isolated, cultured and identified.Exosome was extracted from BM-MSC and observed under transmission electron microscope (TEM). The expression of surface molecular marker CD63 was tested by flow cytometry.Male Sprague-Dawley rats were randomly grouped into sham-operated (Sham), ischemia reperfusion (IR), MSC-treated (IR+ MSC), MSC-derived exosome-treated (IR+ MSC-ex) and fibroblast-derived exosome-treated (IR+ F-ex) groups.The model for ischemia reperfusion injury was constructed.The serum levels of creatinine and blood urea nitrogen (BUN) were tested in each group.The histomorphological changes in of renal tissue samples were examined by hematoxylin and eosin-stained tissue samples.Cellular apoptosis was examined by terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) staining.The expression levels of proinflammatory cytokines interleukin 1β and tumor necrosis factor α were examined by reverse transcription (RT)-PCR.And the expression level of caspase-3 was examined by Western blot.

Rat BM-MSCs were successfully isolated and cultured.Dectection of surface markers revealed high expression levels of CD29 and CD44 and a low expression level of CD34.MSC differentiated successfully into osteoblasts and lipocytes after growing in osteoblast- and lipocyte-inducing media respectively.Typical appearances of exosome were observed under transmission electron microscope.CD63 was positive on flow cytometry.Compared with the IR group, the IR+ MSC and IR+ MSC-ex groups showed low levels of serum creatinine and BUN, mild pathological injury, decreased number of apoptotic cells and low expression of inflammatory factors and caspase-3(IR group, 4 310±616; IR+ MSC group, 2 569±530; IR+ MSC-ex group, 3 144±343, both P<0.05).

Rat BM-MSC-derived exosome protects against ischemia reperfusion injury with decreased inflammatoryresponse and apoptosis in rats.

To explore the efficacy and safety of designed early conversion from calcineurin inhibitor (CNI) to sirolimus (SRL) as major immunosuppressive therapy in renal transplant recipients with stable renal function.

A prospective, open-label and non-randomized control study was performed for 112 renal transplant recipients (3-6 months post-operation) with stable renal function between June 2008 and June 2011.The patients in SRL group (n=57) switched to sirolimus while those in CNI group (n=55) continued CNI.The dosing of mycophenolate mofetil and steroids had no change.They were followed up for at least 24 months to evaluate the acute rejection, patient and graft survival, renal function, estimated glomerular filtration rate (eGFR), blood lipids, blood glucose, liver function and urinary protein at 1, 6, 12 and 24 months after inclusion.Adverse events were also recorded.

The serum creatinine of SRL group decreased significantly after conversion ((89.2±24.7), (87.6±23.8), (86.1±20.4), (86.7±19.7) vs(117.0±16.3) μmol/L, all P<0.05). CNI group showed no improvement of renal function.SRL group had a significantly higher eGFR than CNI group (P<0.05). Among 3 cases of acute rejection, there were 2 in SRL group and 1 in CNI group (P>0.05). Blood lipids in SRL group increased significantly at 1 month after conversion (P<0.05) and reverted back to average level after intervention (P>0.05). SRL group had a drop of hemoglobin level within the normal range.Two patients in SRL group developed hypokalemia and another 2 patients had oral ulcer.They all improved after treatment.During follow-ups, 1 case of mild proteinuria was found in SIR group.Three patients were diagnosed with diabetes (1 in SRL group vs 2 in CNI group).

Early conversion from CNI to SRL as major immunosuppressive therapy in renal transplant recipients with stable renal function further improves renal function.There is no higher rate of acute rejection during follow-up.Elevated blood lipids after conversion may be easily controlled. No other adverseevents are found.

To establish a new detection method for cytomegalovirus(CMV) specific cytotoxic CD8⁺ T cells and examine its proportion and significance in peripheral blood from kidney transplant recipients.

A total of 30 recipients of kidney transplantation from January 2009 to December 2010 for the first time were enrolled. And 10 healthy volunteers were selected as health control group. Tetramer technology was applied. The proportions of CMV antigen specific T cells expressed in each group were detected directly by flow cytometry.

The positive rate of CMV antigen specific CTL, CMV-pp65 specific CD8⁺ T cell was between 0.16%-7.21% in kidney transplant recipients (n=30) and health control group (n=10). The proportions of CMV antigen specific CTL were 2.95%±0.62% in CMV+ group. And it was significantly higher than that in CMV-group (1.17%±0.45%) and health control group (0.65%±0.17%) (P=0.003, 0.006). In CMV+ group, the proportion of CMV antigen specific CTL was 2.95%±0.62% in CMV-pp65 positive phase and decreased significantly to 1.50%±0.32% after turning into negative phase. In CMV+ group (n=15), the proportion of CMV antigen specific CTL was positively related with the number of CMV-pp65 positive cells (Pearson test, r=0.871, P<0.01).

The proportion of specific CTL is an important guide for evaluating and judging the prognosis of CMVinfection. And it may provide rationales for future targeted therapy in kidney transplant recipients.

To explore the generation pattern and clinical relevance of de novo HLA-DQ antibodies in renal transplantation.

A total of 175 primary renal transplant recipients without pre-transplant HLA antibodies were recruited from January 2012 to December 2013. The average follow-up period was 10 (6-18) years. They were divided into control group (n=94) with normal renal graft function (serum creatinine <120 μmol/L); dysfunction group ( n=54) with continued serum creatinine >150 μmol/L over 6 months; and graft loss group ( n=27) with resumed hemodialysis. The sera were collected and screened for de novo HLA antibodies by flow PRA or Luminex mixed beads. And HLA-A, -B, -DR and -DQ specific antibodies were identified by HLA single antigen beads.

Positive de novo HLA antibodies were detected in 48%(26/54)of patients from dysfunction group and in 52%(14/27) from graft loss group, but only in 17%(16/94)from control group (P<0.01). The frequency ofde novo DQ antibodies among patients with any positive HLA antibody was the highest in all three groups (14/16, 24/26 and 14/14). Additionally, the average mean peak fluorescence intensity of DQ antibodies was almost the highest when compared to other antibodies. Moreover, when those with positive HLA antibodies in each group were analyzed, 10/16 in control group were detected to have DQ antibodies alone. However, 17/26 of patients in dysfunction group and 14/14 in graft loss group were detected to have DQ antibodies plus HLA-DR and/or -A, -B antibodies. It indicated that the combined presence of DQ-and non-DQ- antibodies was associated with chronic renal graft failure.

The presence of de novo HLA-DQ antibodies is frequent after renal transplantation. And it is associated with chronic graft failure when co-displaying with other HLA antibodies. The screening and detection of HLA-DQ antibodies after kidney transplantation may aid early warning of donor antigen-specific activation of immune system and subsequent graft dysfunction. Thus it serves as an indicationfor early treatment.

To explore the clinical efficacy and safety of compound digestive enzyme tablet in the treatment of dyspepsia.

A randomized, double-blind, double-dummy, positive drug parallel controlled, multicenter clinical trial was conducted for 203 dyspeptic patients from October 2011 to August 2012.And they were randomized into group A (experimental, n=106) and group B (control, n=97). Group A received 1 tablet of compound digestive enzyme tablet (Bearse) plus 2 analog capsules of compound digestive enzyme thrice daily.And group B had 2 capsules of compound digestive enzyme capsule (Dages) plus 1 analog tablet of compound digestive enzyme thrice daily.The total duration of drug treatment was 2 weeks. There were 3 follow-up visits (W0, W1, W2). The primary endpoint was the total effective rate of all dyspeptic symptoms.

The total efficacy rate of groups A and B were 80.2%(85/106)and 79.4%(77/97) (P>0.05). The adverse effects were 1.9%(2/106)and 4.1%(4/97)in groups A and B (P>0.05). The adverse effects were mild in both groups.

Compound digestive enzyme tablet and capsule are effective and safe for patients with dyspepsia.Andcompound digestive enzymes tablet is comparable to compound digestive enzyme capsule.

To explore the application of absorbable suture continuous catcher (ASCC) method during gastrointestinal anastomosis.

From January 2012 to March 2014, 210 patients with upper gastrointestinal tumors received ASCC method during gastrointestinal anastomosis by single treatment group.They were compared with 300 cases of full-thickness interrupted suture (FTIS) using traditional methods over the same period.Their clinical data were retrospectively analyzed, including anastomotic hemorrhage, leakage and obstruction.

There was neither mortality nor serious abdominal complication.The ASCC group had one case of anastomotic hemorrhage (1/210, 0.05%) while there were 17 cases (17/300, 5.67%) in the FTIS group.The difference was statistically significant with a P value of 0.032.The ASCC group had a lower incidence of anastomotic leakage and obstruction.However, the difference was statistically insignificant with P values of 0.101 and 0.153 respectively.

As compared with the FTIS method, the ASCC method has a lower incidence of gastrointestinal anastomotic hemorrhage and other anastomotic complications. The ASCC method is an ideal suture methodof gastrointestinal anastomosis reinforcement.

To explore the clinical features of rapidly progressive crytogenic organizing pneumonia (COP).

Nine cases of rapidly progressive COP with acute respiratory failure confirmed by lung biopsy at First Affiliated Hospital, Nanjing Medical University from October 2003 to March 2014 were retrospectively reviewed.

There were 5 males and 4 females with an age range of (52.1±11.4) years. The onset of illness was 7-20 days. The major manifestations included cough (n=9), dyspnea (n=9), fever (n=7), hypodynamia (n=6), weight loss (n=3), night sweat (n=2), hemoptysis (n=1), cyanosis (n=9), "velcro" crackles ( n=6) and moist rales (n=3). Blood tests showed elevated levels of erythrocyte sedimentation rate (n=8), C-reactive protein (n=8) and serum ferritin (n=6). The results of blood gas analysis indicated acute respiratory failure in all 9 patients (PaO₂<60 mmHg, 1 mmHg=0.133 kPa). Chest computed tomography showed bilateral and multiple distribution, patchy areas of alveolar consolidation with air bronchograms (n=5), diffuse ground-glass opacity (n=4), tractional bronchiectasis (n=2), pleural effusion (n=2) and spontaneous pneumothorax (n=1). Six cases were cured by conventional dose corticosteroid and another 3 by large-dose methylprednisolone (240 mg/d). Two cases relapsed and then recovered after a second corticosteroid therapy.

The clinical manifestations of rapidly progressive COP show severe inflammatory reactions with a fast progression. Chest computed tomography depicts extensive lesions along with pleural effusion and spontaneous pneumothorax. Conventional dose of corticosteroid is the first choice therapy for rapidly progressive COP. Large-dose methylprednisolone is needed when conventional dose of corticosteroidproves ineffective.

To summarize the disease distribution characteristics of adrenal lesions and analyze the changes of disease spectrum so as to further guide clinical practices.

All clinical data of adrenal lesion patients hospitalized between 1993 and 2013 were collected.Their profiles of gender ratio, visiting age, final diagnosis and year of hospitalization were retrospectively analyzed.

A total of 4 049 patients with adrenal lesions were admitted.There were 2 004 males and 2 045 females with a mean age of 46.82±13.36 years.The number of cases gradually increased year-on-year.And the visiting age was 30-60 years.The annual proportion of adrenal incidentaloma out of total adrenal lesions gradually increased with advancing age.Among them, 3 014 received endocrine function assessments.And there were non-functional lesions (n=1 503, 49.88%), adrenal Cushing's syndrome (n=259, 8.60%), primary aldosteronism (n=665, 22.04%) and pheochromocytoma (n=235, 7.80%) .The proportion of non-functional lesions rose from 20.97% to 54.11% (P<0.05). The proportion of primary aldsteronism declined from 29.84% to 20.43% (P<0.05). And the proportion of adrenal Cushing syndrome changed slightly (P>0.05). The proportion of pheochromocytoma declined from 18.55% to 6.99% (P<0.05).

The number of patients gradually increased over the last two decades.So did the proportion of adrenal incidentaloma.The disease spectrum of adrenal lesions has changed.And the proportion of non-functionallesions increased while that of functional lesions declined.

To explore the major risk factors for pulmonary infection after radical gastrectomy in patients with gastric cancer.

From November 2010 to February 2014, a total of 765 patients undergoing radical gastrectomy at our hospital were divided into 2 groups based upon the presence of postoperative pulmonary infection (n=32, 4.2%) or not (n=733, 95.8%). Their clinicopathological data were retrospectively analyzed by Logistic regressive analysis with a case-control study model.

Comparing with the control group, the patients had longer surgical duration (245.7±66.7 vs 210.9±47.2 min, P<0.01), higher rates of requiring intensive care (12.50% vs 2.86%,P=0.02) and longer post-operative hospital stays (21.9±24.9 vs 14.2±4.2 days, P<0.01) in the postoperative pulmonary infection group.Univariate Logistic regressive analysis found that age ≥60 years, smoking ≥ 400 year by cigarette, diabetes mellitus, chronic obstructive pulmonary disease, proximal or total gastrectomy, combined organ resection, surgical duration ≥240 min, intra-operative blood loss ≥ 300 ml, peri-operative transfusion, transfusion ≥3 unit packed red blood cell, post-operative transfusion and post-operative complications other than pulmonary infections were associated with postoperative pulmonary infection (allP<0.05). Further multivariate analysis identified 4 independent risk factors for pulmonary infection after radical gastrectomy, including diabetes mellitus (OR=4.77, 95%CI: 1.18-19.23), post-operative complications other than pulmonary infections (OR=3.15, 95%CI: 1.25-7.90), intra-operative blood loss ≥ 300 ml (OR=2.63, 95%CI: 1.17-5.90) and post-operative nasogastric tube ≥5 days (OR=2.30, 95%CI: 1.02-5.21).

Correcting the modifiable risk factors may reduce the incidence of pulmonary infection and shortenthe length of hospital stays and costs after radical gastrectomy in patients with gastric cancer.

To explore the gene expression profiles of severe secondary pulmonary tuberculosis patients.

From May 2012 to October 2013, a total of 103 eligible patients with secondary pulmonary tuberculosis were recruited from Institution of Tuberculosis Research of PLA Hospital No. 309. They were divided into severe secondary pulmonary tuberculosis (severe group) (n=57) and mild secondary pulmonary tuberculosis (mild group) (n=46) by the severity of disease .At the same time age-matched healthy controls (n=45) were selected from healthy subjects undergoing physical examination. Whole genome expression profiling was performed with Affymetrix Gene expression chips for 4 cases in severe group, 3 in mild group and 5 in healthy group. Cluster and bioinformatics analysies were performed on differentially expressed genes in severe versus mild group. The remainders of three groups were 53, 43 and 40 cases respectively used for verify the results of gene chip by real-time fluorescence quantitative PCR (RT-PCR). And 20 cases in severe group, 20 in mild group and 8 in control group were used to verify the expression level of jun oncogene (JUN) on behalf of differential expressed genes. Analysis of variance and non-parametric tests were used for statistic difference analysis among three groups.

There were 406 differentially expressed genes for severe and mild groups. There were 264 down-regulated gene and 142 up-regulated ones. The down-regulated genes were predominant. Cluster analysis show the similarity of gene expression profile in the same group .The result confirmed that the gene chip experiments were both repeatable and reliable. According to gene ontology, the differentially expressed genes were mainly involved in such biological processes as immune response, signal transduction, regulation of transcription (DNA-dependent), inflammatory response, antigen processing and presentation and chemotaxis, etc. Pathway analysis showed differentially expressed genes were involved in 22 pathways of immune response and inflammation. The major pathways included B cell receptor signaling, antigen processing and presentation, Toll-like receptor signaling, MAPK signaling and transforming growth factor-beta (TGF-β) signaling.Real-time fluorescence quantitative PCR (RT-PCR) analysis showed that the statistics of optical density forJUN was P<0.001 in severe versus mild group. It was down-regulated in severe group. And the expression ofJUN was conformed with the result of gene expression chip.

The patients of severe group have a larger number of differential expressed genes versus those of mild group. And severe lung tissue damage in severe group may be correlated with differences in geneexpression.

To systemically explore the cellular adhesion signal transduction network of tumor necrosis factor-alpha (TNF-α)-induced hepatocellular carcinoma cells with bioinformatics tools.

Published microarray dataset of TNF-α-induced HepG2, human transcription factor database HTRI and human protein-protein interaction database HPRD were used to construct and analyze the signal transduction network.

In the signal transduction network, MYC and SP1 were the key nodes of signaling transduction. Several genes from the network were closely related with cellular adhesion.Epidermal growth factor receptor (EGFR) is a possible key gene of effectively regulating cellular adhesion during the induction of TNF-α.

EGFR is a possible key gene for TNF-α-induced metastasis of hepatocellular carcinoma.

To explore the therapeutic effects and mechanism of glycyrrhizic acid (GA) on airway inflammation in a murine model of asthma.

A total of 70 female BALB/c mice were randomly divided into 5 groups (n=14 each) of control, asthmatic and three treatments with low, medium and high doses of GA. Mice in the asthmatic and the treatment groups were sensitized and challenged by ovalbumin (OVA). Mice of the treatment groups were injected intraperitoneally with GA (25, 50, 100 mg/kg) 30 min before each OVA challenge. The control mice received an aerosol inhalation of normal saline instead of OVA. Within 24 hours after the last OVA challenge, bronchoalveolar lavage fluid (BALF) was collected for counting total cells and eosinophils (Eos) in other 8 mice in each group. Interleukin (IL)-12p70, IL-10, IL-4 and interferon gamma (IFN-γ ) were measured in BALF by enzyme-linked immunosorbent assay (ELISA). Histological studies of lung were conducted with hematoxylin and eosin staining (HE) and the expressions of CD86, major histocompatibility complex (MHC)-Ⅱ, CD40, OX40 ligand (OX40L) on CD11c⁺ dendritic cells (DCs) in spleens were evaluated with flow cytometry (FCM).

Compared with the control group, the number of total cells and eosinophils was higher in BALF in asthmatic group ((124.3±39.7)×10⁷/L, (26.3±17.2)×10⁷/L vs (55.3±22.8)×10⁷/L, (0.6±0.4) ×10⁷/L), the expressions of IL-10 and IL-4 increased ((49±12, 169±29) ng/L vs (34±4, 89±37) ng/L) and the levels of IFN-γ and IFN-γ/IL-4 ratio decreased in BALF ((122±56 ) ng/L, (0.7±0.4) vs (89±37) ng/L, 2.9±0.8)), the expressions of CD86, MHC-Ⅱ, CD40, OX40L on CD11c⁺ DCs increased in spleens ((38.4±15.7)%, (90.4±3.4)%, (25.4±10.2)%, (29.6±9.9)% vs (18.8±4.4)%, (73.1±11.3)%, (3.8±2.2)%, (5.0±1.6)%) (all P<0.05). Compared with the asthmatic group, total cell counts and the eosinophil numbers significantly decreased by the treatment of GA at a dose of 100 mg/kg ( (62.1±21.7)×10⁷/L, (2.2±1.7)×10⁷/L), the levels of IL-12p70, IL-10, IFN-γ and the ratio of IFN-γ/IL-4 increased ((44±14, 132±13, 208±66) ng/L, (1.8±0.6)) and the level IL-4 decreased (122±38) ng/L. The expressions of MHC-Ⅱ, CD40, OX40L on CD11c⁺ DCs decreased ((75.8±15.9)%, (11.1±5.9)%, (11.8±3.4)%)) (all P<0.05). The asthmatic mice induced an infiltration of inflammatory cells around airways and blood vessels. Administration of GA significantly reduced the infiltration of inflammatory cells in peribronchial areas compared with asthmatic mice especially at a dose of 50 mg/kg 100 mg/kg.

GA effectively ameliorates the airway inflammation of asthma via inhibiting the Th2 responses thoughmodulating the expressions of CD86, MHC-Ⅱ, CD40, OX40L on CD11c⁺ DCs.

To observe the mRNA and protein expression of TLR9, NF-κB and serum level of Th1/Th2 cytokines when stimulated with different concentration of CpG-ODN in STZ-induced diabetic rats.

40 SD male rats were randomly divided into four groups: normal control group (GroupⅠ), type 1 diabetes control group (GroupⅡ), low-dose CpG-ODN treated type 1 diabetes group (Group Ⅲ), high dose CpG-ODN treated type 1 diabetes group (Group Ⅳ).60 μg /kg and 120 μg /kg CpG-ODN were injected to rats of GroupⅢ and Ⅳ when inducing type 1 diabetes model.mRNA and protein levels of TLR9 and NF-κB in spleen and pancreas were detected by fluorescence quantitative PCR and Western blotting.Serum TNF-α, IL-6 and IFN-γ were measured by ELISA.

TLR9 protein expression levels were much higher in spleen than in the pancreas among these four groups.In spleen, TLR9 and NF-κB mRNA and protein level were significantly higher in Group Ⅲand Ⅳthan those in Group Ⅰand Ⅱ, and the expression level was higher in Group Ⅳ than in Group Ⅲ.In pancreas, TLR9 and NF-κB protein level were significantly higher in Group Ⅲand Ⅳthan those in Group Ⅰand Ⅱ, and the expression level was also higher in Group Ⅳ than Group Ⅲ.Serum TNF-α and IFN-γ level in Group Ⅲand Ⅳsignificantly higher than in Group ⅠandⅡ, and serum levels of TNF-α and IFN-γ in Group Ⅳ was higher than in Group Ⅲ.

CpG-ODN activation of TLR9-MyD88 signaling pathways results in dose-effect NF-κB stimulationin STZ-induced diabetic rats.

To observe the effects of insulin-like growth factor-1(IGF-1) on myocardial fibrosis and myocardial cell apoptosis in diabetic rats, so as to clarify the potential benefit of IGF-1 on diabetic cardiomyopathy.

Diabetic rats were induced by caudal injection of STZ, and the diabetic rats were randomly divided into 3 groups: diabetes group, the insulin treatment group and the IGF-1 treatment group.Normal control(NC) group did not receive any therapy.The experiment lasted 10 weeks and the heart weight was recorded to calculate the heart weight index.The serum insulin and IGF-1 level were determined.HE staining and Masson staining were taken on the left ventricular tissue.The cardiomyocyte apoptosis were measured by methods of terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) , the expression levels of IGF-1R in cardiomyocytes were measured by immunohistochemical and Western blot.

Compared with diabetes group, the heart weight index of the insulin treatment group and the IGF-1 treatment group were significantly decreased. Serum levels of IGF-1 in diabetes group were significantly decreased compared with NC group, and the levels were significantly increased in IGF-1 treatment group. The results of HE and Masson staining showed that compared with NC group, diabetes rats' myocardial tissue were damaged .Myocardial tissue pathological changes of rats were significantly improved after insulin and IGF-1 treatment. The result of TUNEL indicant cardiomyocyte apoptosis index in diabetes group was obviously increased than the NC group, and that in IGF-1 treatment group were decreased compared with diabetes group. IGF-1R levels in diabetic rats myocardial tissue were rised than the NC group, it was declined in IGF-1 treatment group, but in insulin treatment group the levels had no obvious change.

IGF-1 can inhibite the myocardial fibrosis and cardiomyocyte apoptosis. It maybe have the possibleeffect on the treatment of diabetic cardiomyopathy.

To compare the efficiency of 1.9 μm Vela Laser versus high-frequency electronic knife in the treatment of digestive tract big polyps.

A total of 30 patients with big polyps (>2 cm in diameter) hospitalized from May 2013 to November 2013 were randomly divided into two groups to receive 1.9 μm Vela Laser or high-frequency electric knife respectively.The sites of polyp were at stomach (n=4), duodenum (n=4), sigmoid colon (n=12) and large intestine (n=10). Surgical duration, outcomes, intra-operative and post-operative complications, lesion remnant and relapse possibilities were systematically reviewed to compare the efficiency of these two techniques.

The average operating duration of high frequency electronic knife treatment was shorter than 1.9 μm Vela Laser treatment (20.9±1.4 vs 27.6±1.2 min). For those with high frequency electronic knife, there were obvious wound bleeding ( n=2); intra-operative perforation (n=1) and abdominal discomforts within 24 hours post-surgery (n=3). In contrast, none of those with 1.9 μm Vela Laser suffered the above complications.All wounds were well-healed after 6 months, except for 2 patients with high frequency electronic knife leaving an apparent scar at wound site (without significant stricture). One case had residual lesion during the multi-point biopsy.

By avoiding such side-effects as bleeding, perforation and stomachache, 1.9 μm Vela Laser possesses many advantages over high frequency electronic knife.However, the former technique is in its early development period and some bottlenecks such as a longer operating duration should be urgentlyaddressed.

Peri-prosthetic aortic valve leakage (PVL) is one of the most common serious complications after cardiac surgical prosthetic valve replacement. In the past, surgical thoracotomy was used to repair paravalvular leakage, or valve replacement again, but the trauma was large, the risk was high, and it was easy to repeat residual leakage. In recent years, transcatheter occlusion of PVL has become another important alternative treatment^[1,2,3]。 However, this method has no special occluder, the operation is relatively complicated, and there are complications such as aggravation of postoperative hemolysis. In this study, a new polyester-free second generation vascular plug occluder (AVP Ⅱ) was applied to aortic valve PVL, which is easier to operate and can effectively reduce complications.

Exhaled nitric oxide (FeNO) detection is a new non-invasive airway inflammation detection technique developed in recent years. It has the advantages of quantitative, non-invasive, simple and safe, and has high specificity and sensitivity^[1]。 The American Thoracic Society/European Society of Respiratory Diseases (ATS/ERS) published standard practice for testing FeNO in 2005^[2]and reaffirmed the value of FeNO as a marker of airway inflammation in 2009 and 2011^[3,4]。 Many countries have successively carried out testing of FeNO. In 2006, the Global Asthma Initiative (GINA) included FeNO in asthma management protocols^[5]In 2008, the Respiratory Branch of Chinese Medical Association listed it in the Guidelines for the Prevention and Treatment of Bronchial Asthma^[6]。 The application progress of FeNO in the diagnosis and treatment of respiratory diseases is summarized as follows.

A 56-year-old male was seen for "abdominal pain accompanied by fever for 15 d, cough and shortness of breath". The patient had multiple skin nodules for more than 30 years, showing a trend of gradual aggravation, and had a history of hypertension for more than 10 years. The patient had a son and a brother with multiple nodules in the skin and was diagnosed as neurofibroma type I. Physical examination: There were extensive tumor-like masses of varying sizes on the skin of the whole body, coffee-like spots widely distributed on the epidermis, ecchymosis under the skin, no optic nerve atrophy, no papilledema and iris hamartoma (Lish nodule) nodules were found on fundus examination; A mass about 10 cm ×15 cm in size can be accessed in the left upper abdomen, with poor boundary, and positive percussion pain in the left kidney area. The digital image shows: thoracic scoliosis deformity, and multiple nodules of different sizes are seen in the soft tissues of the chest wall before and after the nodes on both sides; CT examination showed that an oval cystic mass of 7.0 cm ×7.7 cm ×8.9 cm was seen in the left upper abdominal cavity. The mass was closely related to the body and tail of the pancreas, and the boundary was unclear. There was no enhancement in the mass, uneven thickness of the mass wall, obvious enhancement of the cyst wall, separation, partial wall and separation shadows were seen in the mass. The boundary between the mass and the adjacent gastric cavity, spleen, left kidney and adrenal gland was clear. The left kidney was slightly moved downward under compression, and the boundary between the splenic artery and left renal vein was clear (FIG. 1A, lb); No enlarged lymph nodes were seen retroperitoneally, and multiple soft tissue density nodules and masses were seen subcutaneously in the abdomen, with mild enhancement (Figure 1C)； Multiple soft tissue density nodules and masses were seen under the scalp, and small round low density shadows were seen in the skin, with no obvious enhancement (Figure 1D)； The patient underwent abdominal mass resection, small intestinal mass resection, and subcutaneous nodule resection of abdomen and trunk. Postoperative pathology: (1) Abdominal subcutaneous and trunk nodules: HE staining showed that the tumor was composed of loosely arranged spindle cells, with few and indifferent cytoplasm, slender and wavy nucleus, and the cells were embedded in fibrous matrix (Figure 2)； (2) Adrenal gland: 1 gray-red gray-yellow mass, 7.0 cm ×7.7 cm ×8.9 cm in size, with calcification of cyst wall; HE: The tumor tissue cells are organ-like arrangement, with abundant blood vessels, moderate cell abnormality, intact envelope, calcification, most of the tumors in the envelope are bleeding, necrosis, and cholesterol crystals (Figure 3)； Immunohistochemical staining results: S-100 (+ +), synaptophysin (+ +), chromaffin (–), neuron-specific enolase (–), cytokeratin 7 (–), carcinoembryonic antigen (–), cell proliferation marker Ki-67 (–), tumor suppressor gene P53 (–), cytokeratin 18 (–); (3) Tumor of small intestinal wall: a piece of gray-white nodular tissue with mucosa, the size is 1.3 cm ×1.3 cm ×1.0 cm, and the mucosa area is 1.6 cm ×0.4 cm; Immunohistochemical staining results: neural marker acid binding protein S – 100 (–), oncogene product CD117 (+ +), endothelial cell marker CD34 (+ +), smooth muscle actin (+ +), Ki – 67 (–) (Figure 4，Figure 5)。 The diagnosis was as follows: neurofibroma type I with giant pheochromocytoma and small intestinal stromal tumor.

A 4-year-old child came to the Department of Ophthalmology of Beijing Children's Hospital for 3 years of photophobia in both eyes, with blepharospasm, polydipsia and polyuria, and lagging growth and development. No one else in the family had similar symptoms. The child had a full-term cesarean section, growth retardation, irritability, irritability, daily water consumption 1 000~1 200 ml. Full body examination: height 78 cm, weight 9.1 kg, square skull, mild valgus of bilateral costal margins, bracelet sign (+), bracelet sign (+) (Figure 1), neurological pathology (–).