A review of lipid accumulation by oleaginous yeasts: Culture mode.

Microbial lipids have attracted considerable interest owing to their favorable environmental sustainability benefits. In laboratory-scale studies, the factors impacting lipid production in oleaginous yeasts, including culture conditions, nutrients, and low-cost substrates, have been extensively studied. However, there were several different modes of microbial lipid cultivation (batch culture, fed-batch culture, continuous culture, and other novel culture modes), making it difficult to comprehensively analyze impacting factors under different cultivation modes on a laboratory scale. And only few cases of microbial lipid production have been conducted at the pilot scale, which requires more technological reliability assessments and environmental benefit evaluations. Thus, this study summarized the different culture modes and cases of scale-up processes, highlighting the role of the nutrient element ratio in regulating culture mode selection and lipid accumulation. The cost distribution and environmental benefits of microbial lipid production by oleaginous yeasts were also investigated. Our results suggested that the continuous culture mode was recommended for the scale-up process because of its stable lipid accumulation. More importantly, exploring the continuous culture mode integrated with other efficient culture modes remained to be further investigated. In research on scale-up processes, low-cost substrate (organic waste) application and optimization of reactor operational parameters were key to increasing environmental benefits and reducing costs.

A putative PKA phosphorylation site S227 in MoSom1 is essential for infection-related morphogenesis and pathogenicity in Magnaporthe oryzae.

In the rice blast fungus Magnaporthe oryzae, the cAMP signalling pathway plays a critical role in regulating leaf surface recognition and the initiation of appressorium development. Direct downstream targets of the cAMP signalling pathway are, however, not well-characterised. The MoSom1 protein functions downstream of the cAMP dependent protein kinase A (cAMP-PKA) and is essential for infection-related morphogenesis and pathogenicity. In this study, we show that mutation of a putative PKA phosphorylation site in MoSom1 is essential for its role in appressorium differentiation and pathogenicity in M. oryzae. Mutation of serine 227 in MoSom1 by deletion or serine (S) substitution to alanine (A), valine (V) or tyrosine (Y), resulted in defects of conidiation, appressorium-like structure formation and fungal pathogenicity. Western blot analysis confirmed that S227 in MoSom1 is a putative PKA phosphorylation site. Furthermore, a ΔMosom1 mutant showed reduced expression of PMK1 and was defective in Pmk1 phosphorylation, indicating that the Pmk1 mitogen-activated protein kinase (MAPK) acts downstream of MoSom1 in M. oryzae. We conclude that the cAMP-PKA pathway may regulate the Pmk1 MAPK pathway through MoSom1 during rice infection by the blast fungus. TAKE AWAYS: S227 is crucial for MoSom1 function in M. oryzae. S227 in MoSom1 was identified as a putative PKA phosphorylation site in M. oryzae. S227 is essential for infection-related morphogenesis and pathogenicity in M. oryzae.

Endobronchial Ultrasound Combined with Clinical Data for Predicting Malignant Peripheral Pulmonary Lesions.

INTRODUCTION: This study was to develop a simple model for predicting malignancy of peripheral pulmonary lesions (PPLs) based on endobronchial ultrasonography (EBUS) and clinical findings. METHODS: Patients who had EBUS for PPLs were analyzed and compared on the EBUS imaging characteristics and clinical data. The malignancy prediction model was established by the logistic equation of probability of malignant PPL based on the data of 135 patients. The model was tested on an additional 50 patients for efficiency. RESULTS: Among 135 prospectively enrolled patients, 77 (57%) patients had malignant and 58 (43%) had benign lesions with the size of 36.5±19.9 mm. Univariate analysis demonstrated a significant (P<0.05) difference in the serum CEA (borderline 15 µg/mL) and smoking history between malignant and benign lesions but a non-significant (P>0.05) difference in age (50 years as the cutoff value) and history of extra-thoracic malignancies. Logistic analysis of multiple factors showed that smoking history, serum CEA, borderline, air bronchogram, heterogeneous echo, and anechoic areas were significant (P<0.02) risk factors for malignant lesions. The malignancy prediction model was established by the logistic equation of probability of malignant PPL (P) = l/[l+e-Z], where Z=-2.986+1.993X1+2.293X2+l.552X3+1.616X4-2.011X5+1.718X6, e is the base of the natural logarithm, X1 is the smoking history, X2 is the serum CEA, X3 is the borderline, X4 is the heterogenicity, X5 is the air bronchogram, and X6 is the anechoic area. The receiver operating characteristic curve had an area under the curve (AUC) of 0.926 (95% confidence interval: 0.883-0.969). The sensitivity, specificity, and accuracy were 88.2% (30/34), 75.0% (12/16), and 92.0% (46/50), respectively, for the logistic equation to predict the malignancy. CONCLUSION: Endobronchial ultrasonography is a safe and practical method, and the model combining EBUS and clinical data can accurately predict the malignancy of peripheral pulmonary lesions.

Draft Genome Sequence of Pseudarthrobacter sp. Strain AG30, Isolated from a Gold and Copper Mine in China.

Here, we report the features and draft genome sequence of Pseudarthrobacter sp. strain AG30, isolated from the Zijin gold and copper mine in China. The genome size of Pseudarthrobacter sp. AG30 was 4,618,494 bp, with a G+C content of 66.2%. Interesting genes and operons putatively conferring resistance to copper and arsenic were identified.

Comparison of fluticasone propionate with budesonide administered via nebulizer: a randomized controlled trial in patients with severe persistent asthma.

BACKGROUND: This study compared the efficacy and safety of fluticasone propionate (FP) inhalation n solution with budesonide (BUD) suspension for inhalation administered via nebulizer, in Chinese adult patients with severe, persistent asthma. METHODS: This was a multicenter, randomized, active-controlled, single-blind, parallel-group study, conducted at 26 clinical sites in China. Participants were randomized 1:1 to FP nebules 1 mg twice daily or BUD 2 mg twice daily via nebulizer for 12 weeks. RESULTS: A total of 317 adult patients were randomized. The primary endpoint was mean change in morning peak expiratory flow (PEF) over weeks 1-12 from baseline, and analyzed in the ITT (n=315) and PP populations (n=283). Week 12 PEF increase from baseline was 26.7 L/min (14.1%) and 28.0 L/min (15.3%) in the ITT population, and 29.1 L/min (15.7%) and 30.1 L/min (16.2%) in the PP population, in the FP and BUD groups, respectively; all improvements were of clinical significance. Lower limits of the two-sided 95% CIs for the least squares (LS) mean treatment difference (FP minus BUD) were -12.19 L/min (ITT) and -12.95 L/min (PP), both above the pre-specified non-inferiority criteria -12.00 L/min and not clinically meaningful. There was no significant difference in the week 12 mean FEV1 increase between the FP and BUD groups (0.237 L/16.79% vs. 0.236 L/17.73%). Lower limits of the 95% CIs for LS mean treatment difference in morning PEF change from baseline over weeks 1-4 in a post hoc analysis were -10.41 and -11.96 L/min in the ITT and PP populations respectively; both above -12.00 L/min. A review of safety data indicated that rates of AEs, SAEs, and drug-related AEs were similar between two groups. CONCLUSIONS: The 12-week treatment of FP inhalation solution administered via nebulizer is safe and effectively for treating severe, persistent asthma in Chinese patients over 12 week.

Functional chitosan-stabilized nanoscale zero-valent iron used to remove acid fuchsine with the assistance of ultrasound.

Chitosan-stabilized nanoscale zero-valent iron (CS-nZVI) was prepared and used for the removal of acid fuchsine (AF) from aqueous solution with the assistance of ultrasound. More than 98.9% of AF was removed using CS-nZVI, aged CS-nZVI (exposed to air for 2 months), while only 14.6% removal efficiency was achieved after 15 min by chitosan alone with the assistance of ultrasound. Scanning electron microscopy (SEM) confirmed that chitosan polymers were arranged in a homocentric layered structure. Thus, the polymer can prevent the aggregation of nZVI and increase their anti-oxidation capacity. X-ray diffraction (XRD) also suggested that the chitosan used in synthesis may protect nZVI nanoparticles from air oxidation. Different factors impacting on the removal of AF using CS-nZVI showed that the reduction increased when dosage and temperature increased, but decreased when pH and initial concentration rose. Kinetic studies revealed that the removal of AF fitted well to the pseudo-first-order model. The apparent activation energy was 55.34 kJ/mol, indicating a chemically controlled reaction. Finally, the application of CS-nZVI in dyeing wastewater led to a removal efficiency of 99% of AF, while the reuse test confirmed that AF's removal efficiency declined from 99.6 to 39.3% after seven cycles.

Multifunctional kaolinite-supported nanoscale zero-valent iron used for the adsorption and degradation of crystal violet in aqueous solution.

Kaolinite-supported nanoscale zero-valent iron (K-nZVI) was successfully synthesized as a multifunctional composite and used for the degradation of crystal violet (CV). The presence of kaolinite not only decreased the aggregation of zero-valent iron nanoparticles (nZVI) with maintenance of reactivity, but also facilitated reaction by increasing the local concentration of CV in the vicinity of nZVI as an adsorbent. This was confirmed by scanning electron microscopy (SEM) and batch experiments, which showed that 97.23% of CV was removed using K-nZVI, while only 78.72% and 39.22% of CV were removed using nZVI and kaolinite after 30 min, respectively. Different factors impacting on degradation of CV were investigated as well. On the basis of these results, a removal mechanism was proposed including: (i) prompt adsorption of CV to the K-nZVI phase, and (ii) reduction of CV by Fe(0) on K-nZVI. Furthermore, different adsorption and reduction kinetics were employed to examine the removal process of CV, where a better fit with the pseudo-second-order model for adsorption and pseudo-first-order model for reduction process was observed, and reduction was the rate limiting step. In addition, isotherm and thermodynamic parameters were evaluated for a specific study of the important adsorption step. Finally, the application of K-nZVI to treat wastewater showed the removal efficiency higher than 99.9%.

[Non-real-time endobronchial bronchoscopy ultrasound assisted transbronchial lung biopsy in diagnosing peripheral pulmonary lesions].

OBJECTIVE: To evaluate the role of non-real-time endobronchial bronchoscopy ultrasound(EBUS) assisted transbronchial lung biopsy (TBLB) in diagnosing peripheral pulmonary lesions (PPL). METHODS: One hundred and five patients [68 males and 37 females, mean age (59 ± 12) years, ranged from 39 - 81 years] with PPL confirmed by computered tomography (CT) were recruited in this study between June 1st 2011 and March 1st 2012. All cases received bronchoscopy examinations and presented with roughly normal results. Fifty-four cases received EBUS examinations. For peripheral lesions with accessible EBUS images, blind biopsy was performed with biopsy forceps through pathways of the ultrasonic probe after the retreat of the probe. In those cases without accessible EBUS images, blind biopsy was performed based on the localization by image data. The other 51 cases without EBUS testing underwent blind biopsy on the localization by image data. Positive rates of pathological diagnosis of the 2 groups were compared. Analysis was by χ(2)-test. RESULTS: In 54 patients who received EBUS examinations, 76% (41/54) of PPLs were detected performed by EBUS. The positive rate of the EBUS assisted TBLB group was 67% (36/54), compared with 45% (23/51) in the general TBLB group. There was a better diagnostic rate (P < 0.05) in the EBUS assisted TBLB group than the general TBLB group. Thirteen patients without accessible EBUS images obtained negative pathological results. The diagnosis rate of EBUS assisted TBLB on lesions with ≤ 30 mm minimum diameter was 44% (8/18), lower than 78% (28/36) on lesions with > 30 mm minimum diameter (P < 0.05). In terms of diagnosis rate on lesions with ≤ 30 mm minimum diameter, EBUS assisted TBLB was 44% (8/18), higher than 12% (2/17) of TBLB alone (P < 0.05). As for lesions with > 30 mm minimum diameter, diagnosis rate of EBUS assisted TBLB was 52% (28/54) and TBLB alone was 41% (21/51), representing insignificant difference (P > 0.05). In the EBUS assisted TBLB group, we performed 269 blind biopsies, with an average of 4.8 times per case, whereas the general TBLB group required 398 times, with an average of 7.8 times per case. EBUS assisted TBLB decreased the operation times of blind biopsy (P < 0.05) to acquire adequate and appropriate specimen. Complications of biopsy occurred in this study included slight haemoptysis (61/105, 58.1%), chest pain (25/105, 23.8%) and pneumothorax (2/105, 1.9%). Patients with these complications recovered spontaneously without special managements. CONCLUSIONS: Non-real-time EBUS assisted TBLB could improve diagnostic positive rate without increasing operational risk. In most cases, the blind biopsy did not succeed if EBUS failed to detect the lesions. The success rate of non-real-time EBUS assisted TBLB was related to the minimum diameter of PPL. In terms of diagnosis rate on lesions with ≤ 30 mm minimum diameter, EBUS assisted TBLB was higher than TBLB alone. As for lesions with >30mm minimum diameter, there was no significant difference in the diagnosis rate between these 2 groups. EBUS assisted TBLB decreased the times of blind biopsy process (P < 0.05) to obtain adequate and appropriate specimen.

[Quantitative analysis of the ultrasonogram and histological findings in normal airway wall].

OBJECTIVE: To find out the correlation between endobronchial ultrasonography (EBUS) images and histologic findings in normal bronchial wall via quantitative analysis of the airway wall thickness and the layer thickness. METHODS: From July 1st to December 31th in 2010, patients underwent lobectomy performed endobronchial ultrasonography (EBUS) before surgery and frost pathological examination after surgery. The layer thickness of EBUS and pathological images were measured. Bland-Altman plots were used to analyze the agreement between EBUS measurements and pathological measurements. RESULTS: Twenty-one patients were enrolled in the study. Five layers of the wall were distinguished at the ultrasonogram. Starting on the luminal side, the first, third and fifth layer (L1, L3, L5) were hyperechoic while the second, fourth layer (L2, L4) were hypoechoic. The wall thickness with good agreement was almost equal between the 2 kinds of images (1.877:1.745). L1 thickness was lager than the mucosa thickness (0.275:0.164). L2 thickness was smaller than the submucosa thickness (0.100:0.202). L1 + L2 thickness was almost equal to the thickness of mucosa and submucosa layer (0.375:0.366). The Bland-Altman plots showed poor agreement between the L1, L2 thickness and the mucosa thickness, the submucosa thickness while good agreement between the L1 + L2 thickness and the thickness of mucosa and submucosa layer. L3 thickness was lager than the inner perichondrium thickness (0.241:0.075), and L4 thickness was smaller than the cartilage layer thickness (0.655:0.811). L3 + L4 thickness was almost equal to thickness of the inner perichondrium and the cartilage layer (0.895:0.887). L5 thickness was almost equal to thickness of the outer perichondrium and the connective tissue outside the cartilage layer (0.533:0.491). The Bland-Altman plots showed poor agreement between the L3, L4 thickness and the inner perichondrium thickness, cartilage layer thickness, while good agreement between L5, L3 + L4, L3 + L4 + L5 thickness and the corresponding indexes. CONCLUSIONS: There is a five-layer structure on the bronchial EBUS image including the first layer at the luminal side corresponding to the mucosa and inner part of the submucosa; the second layer corresponding to the outer part of submucosal tissue; the third layer corresponding to the inner perichondrium and the inner part of the cartilage; the fourth layer corresponding to the outer part of cartilage; the fifth layer corresponding to the outer perichondrium and the connective tissue outside the cartilage layer.

Radial probe endobronchial ultrasound scanning assessing invasive depth of central lesions in tracheobronchial wall.

BACKGROUND: Patients with central tracheobronchial benign or malignant lesions who have not recieved surgical treatment can be treated by interventional techniques, such as laser, afterloading radiotherapy, cryotherapy, photodynamics treatment, radiofrequency ablation and stenting, etc. The accuracy of the invasive depth of central lesion in tracheobronchial wall plays an important role in making interventional treatment plan. This study used radial probe endobronchial ultrasound (RP-EBUS) scanning to evaluate the accuracy of the invasive depth of central lesions in tracheobronchial wall, and the influence of RP-EBUS scanning in treatment plan making and guidance. METHODS: This was a prospective study of consecutive patients with central tracheobronchial lesions found by CT or bronchoscopy. We performed EBUS scanning after common bronchoscopy under local anesthesia. A radial ultrasonic probe (2.0 mm in diameter with 20-MHz frequency) with a balloon sheath was introduced through the 2.8-mm-diameter channel of a flexible bronchoscope. The balloon at the tip of the probe was inflated with distilled water until coupling with the airway wall under endoscopic control. The circular image of EBUS, which revealed the layered structure of the tracheobronchial wall, could be achieved. RESULTS: Total of 125 patients were enrolled in the study. Thirty patients underwent surgical operation and pathologically proved the RP-EBUS diagnosis accuracy of tumor invasive depth in tracheobroncial wall was 90% (27/30), sensitivity and specificity were 88.89% (24/27) and 100% (3/3), respectively. In response to EBUS images, 40 approaches were altered or guided: lymph-node metastasis and compressive lesions was diagnosed by EBUS-guided transbronchial needle aspiration (TBNA) (n = 8); Lesions ablation with laser or electricity were stopped when EBUS demonstrated close range with vessels or perforation possibility (n = 13), stents size were changed (n = 14), operation was canceled (n = 3) and foreign body was removed (n = 2). No complication associated with the use of EBUS was observed. CONCLUSION: RP-EBUS can be a useful tool in assessing the central lesion invasive depth to the tracheobronchial wall.

[Differential diagnosis of malignant and benign peripheral pulmonary lesions based on two characteristic echo features of endobronchial ultrasonography].

OBJECTIVE: To assess the feasibility of endobronchial ultrasonography (EBUS) in the differential diagnosis of malignant and benign lesions based on the two characteristic echo features of malignancy. METHODS: EBUS images from 102 patients undergoing bronchoscopy for peripheral lung lesions were analyzed. The sensitivity and specificity were determined for each echo feature, namely the halo sign and low-level echoes that indicated malignancy, or their combination in diagnosing malignant and benign lesions. RESULTS: Low-level echoes showed a sensitivity of 89.46% and a specificity of 83% in the diagnosis of malignancy, both higher than those of the halo sign (69.51% and 65%, respectively). The presence of either of the two echo features had a diagnostic sensitivity of 94.6% for malignant lesions, and the coexistence of the two features had a specificity of 93% for a diagnosis of malignant lesions. CONCLUSION: EBUS is a useful adjunctive modality for lung cancer diagnosis, especially in cases where peripheral lung lesions are invisible in conventional bronchoscopy.

[Preparation of a monoclonal antibody against polyhedrin of Ectropis obliqua nucleopolyhedrovirus].

To develop a method based on immunoreactions for detection of Ectropis obliqua Nucleopolyhedrovirus (EoNPV), the polyhedra of the virus were purified and used to immunize the mouse BALB/c. The spleen cells from the immunized mice were then fused with the myeloma cell line Sp2/0. A hybridoma cell line which can stably secrete the monoclonal antibody against EoNPV was achieved by using indirect ELISA screening and cloning methods, and was named as 7D3. Meanwhile, the polyhedrin gene was cloned from EoNPV and expressed in E. coli. Western blotting analysis showed that the monoclonal antibody prepared from 7D3 could specifically react with the recombinant polyhedrin. An indirect ELISA method based on this monoclonal antibody for detecting EoNPV in infected tea looper was developed.

Removal of methyl orange from aqueous solution using bentonite-supported nanoscale zero-valent iron.

Zero-valent iron (ZVI) nanoparticles tend to agglomerate, resulting in a significant loss in reactivity. To address this issue, synthesized bentonite-supported nanoscale zero-valent iron (B-nZVI) was used to remove azo dye methyl orange (MO) in aqueous solution. Batch experiments show that various parameters, such as pH, initial concentration of MO, dosage, and temperature, were affected by the removal of MO. Scanning electron microscopy (SEM) confirmed that B-nZVI increased their reactivity and a decrease occurred in the aggregation of iron nanoparticles for the presence of bentonite (B). Using B-nZVI, 79.46% of MO was removed, whereas only 40.03% when using nZVI after reacting for 10 min with an initial MO concentration of 100 mg/L (pH=6.5). Furthermore, after B-nZVI reacted to MO, XRD indicated that iron oxides were formed. FTIR showed that no new bands appeared, and UV-vis demonstrated that the absorption peak of MO was degraded. Kinetics studies showed that the degradation of MO fitted well to the pseudo first-order model. A degradation mechanism is proposed, including the following: oxidation of iron, adsorption of MO to B-nZVI, formation of Fe(II)-dye complex, and cleavage of azo bond. Finally, the removal rate of MO from actual wastewater was 99.75% when utilizing B-nZVI.

[Application of high-frequency electrocautery combined with high pressure balloon expansion in treatment of benign airway obstruction].

OBJECTIVE: To study the value of high-frequency electrocautery combined with high-pressure balloon expansion in the treatment of benign airway obstruction. METHODS: Twelve patients with complete airway obstruction received treatment with high-frequency electrocautery and high-pressure balloon expansion, and 6 months of follow-up was performed. RESULTS: After the initial treatment, the rate of airway obstruction relief exceeded 50% in 8 patients, who showed lung recruitment and an increased dyspnea index, suggesting total effectiveness; one patient showed mild improvement with airway stenosis relief by less than 50% (the upper-left lobe remained obstructed). Stents were implanted eventually in 7 patients for airway maintenance. Two patients showed relief of the airway obstruction by over 50% after the initial treatment, but lung recruitment or dyspnea relief failed to be achieved, suggesting non-response to the treatment. The airway failed to be found in 1 case (who received 5 previous operations of ablation and dilatation), which was considered a non-response case. CONCLUSION: High-frequency electrocautery combined with high pressure balloon expansion is clinically effective with few complications in some cases of benign complete airway stenosis. The possibility of airway stenosis and even obstruction should be considered in patients with tuberculosis during the treatment.

[Pathological basis of air bronchogram examined by endobronchial ultrasound in patients with peripheral lung cancer].

BACKGROUND AND OBJECTIVE: Air bronchogram were visible in endobronchial ultrasound images of benign and malignant lesions. The aim of this study is to analyze the endobronchial ultrasound images of different characteristics of air bronchogram and clinical significance combined with pathologic section. METHODS: From June 1, 2005 to December 30, 2008, 92 patients were found pulmonary peripheral lesions by X-ray and CT examinations, and confirmed that lesions were located lower to the segmental bronchial orifice by flexible bronchoscopy examinations. These patients were examined by radial endobronchial ultrasound probe afterwards. RESULTS: Seventy-eight patients clarified with benign or malignant diagnosis of lesions were analyzed, among whom, 22 of 47 (46.8%) were of malignant lesion without air bronchogram, 22 patients in 25 of lesions without air bronchogram were confirmed malignant (88%), among whom, 66.7% (2/3), were of small cell lung cancer, 43.9% (18/41) were of non-small cell lung cancer, and 50% (5/10) were of poorly differentiated adenocarcinoma, no sign of air bronchogram was found in corresponding pathological sections. Among those patients with malignant lesion, 51.1% (24/47) were of irregular air bronchogram. For patients with malignant lesions, irregular air bronchogram was most commonly found in adenocarcinoma, 55.2% (16/29) of pathological sections showed signs of air bronchogram, similar sign was also found in 2 patients with moderately differentiated adenocarcinoma and 1 patient with poorly differentiated adenocarcinoma. The total percentage of patients without air bronchogram and with irregular air bronchogram was 97.9% (46/47), only one of them (with medium differentiated adenocarcinoma) showed regular air bronchogram (1.3%). 80.6% (25/31) patients with benign lesion showed regular sign of air bronchogram distributed in the shape of concentric circles, the percentage of patients without air bronchogram or with irregular air bronchogram was both 3.8% (3/31). CONCLUSION: When there's no air bronchogram, or irregular air bronchogram sign found by endobronchial ultrasound examination, it is highly indicated that there is a malignant lesion, while there's regular sign of air bronchogram distributed in the shape of concentric circles, it is likely to be a benign lesion.

[Morphology and edge analysis of endobronchial ultrasound images in 47 patients with pulmonary malignant lesions].

BACKGROUND AND OBJECTIVE: By analyzing morpholoy and edge of endobronchial ultrasound (EBUS) images in 47 patients with pulmonary malignant lesions. The aim of this study is to explore the role of EBUS in the diagnosis of pulmonary malignant lesions. METHODS: From June 1, 2005 to June 30, 2006, EBUS analysis was performed in a proportion of patients with malignant or benign lesions. RESULTS: In total 78 patients of confirmed diagnosis, 47 cases were confirmed malignant lesions, and 31 were benign lesions, male 56 cases, female 22 cases, age span from 21-80 years (58.01 +/- 13.20). The statistics of the lesion edge imaging in 78 patients showed that different position and angle did not affect the morphous of the edge; analysis of the relationship between the size of the lesion and the morphous of the edge shows that the size of the lesion does not affect the morphous of the edge; the comparison between lesions of benign and malignant shows that a clear edge is the major feature of malignant lesions, indicating certain value in diagnosis; comparison between the shape of the lesion and the property of the lesion shows no relationship between them; compared to conventional bronchoscopy operations, the EBUS operation consumes approximately 10 min, and no related complication was found. CONCLUSION: Clear morphous of the edge in EBUS is a feature of the malignant lesion, and EBUS has certain value in the diagnosis of the malignant lesions.

Expression of recombinant AccMRJP1 protein from royal jelly of Chinese honeybee in Pichia pastoris and its proliferation activity in an insect cell line.

Major royal jelly protein 1 (MRJP1) is the most abundant member of the major royal jelly protein (MRJP) family of honeybee. Mature MRJP1 cDNA of the Chinese honeybee (Apis cerana cerana MRJP1, or AccMRJP1) was expressed in Pichia pastoris. SDS-PAGE showed that recombinant AccMRJP1 was identical in molecular weight to the glycosylated AmMRJP1 from the Western honeybee (Apis mellifera). Western blots probed with anti-AccMRJP1 antibody demonstrated that recombinant AccMRJP1 and soluble protein of the Western honeybee RJ (AmSPRJ) contained immunoreactive MRJP1. The 57 kDa protein in AmSPRJ contained an N-terminal amino sequence of N-I-L-R-G-E, which is identical to that previously characterized in AmMRJP1. The molecular weight of recombinant AccMRJP1 was decreased from 57 to 48 kDa after deglycosylation, indicating that AccMRJP1 was glycosylated. The recombinant AccMRJP1 significantly stimulated Tn-5B-4 cell growth, similar to AmSPRJ and fetal bovine serum, and affected cell shape and adhesion to the substrate.