MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis.

BACKGROUND: MicroRNAs (miRNAs) regulate gene expression and play a critical role in cancer physiology. However, there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer (GC). AIM: To investigate the role and molecular mechanism of miRNA-145-5p (miR145-5p) in the progression of GC. METHODS: Real-time polymerase chain reaction (RT-PCR) was used to detect miRNA expression in human GC tissues and cells. The ability of cancer cells to migrate and invade was assessed using wound-healing and transwell assays, respectively. Cell proliferation was measured using cell counting kit-8 and colony formation assays, and apoptosis was evaluated using flow cytometry. Expression of the epithelial-mesenchymal transition (EMT)-associated protein was determined by Western blot. Targets of miR-145-5p were predicated using bioinformatics analysis and verified using a dual-luciferase reporter system. Serpin family E member 1 (SERPINE1) expression in GC tissues and cells was evaluated using RT-PCR and immunohistochemical staining. The correlation between SERPINE1 expression and overall patient survival was determined using Kaplan-Meier plot analysis. The association between SERPINE1 and GC progression was also tested. A rescue experiment of SERPINE1 overexpression was conducted to verify the relationship between this protein and miR-145-5p. The mechanism by which miR-145-5p influences GC progression was further explored by assessing tumor formation in nude mice. RESULTS: GC tissues and cells had reduced miR-145-5p expression and SERPINE1 was identified as a direct target of this miRNA. Overexpression of miR-145-5p was associated with decreased GC cell proliferation, invasion, migration, and EMT, and these effects were reversed by forcing SERPINE1 expression. Kaplan-Meier plot analysis revealed that patients with higher SERPINE1 expression had a shorter survival rate than those with lower SERPINE1 expression. Nude mouse tumorigenesis experiments confirmed that miR-145-5p targets SERPINE1 to regulate extracellular signal-regulated kinase-1/2 (ERK1/2). CONCLUSION: This study found that miR-145-5p inhibits tumor progression and is expressed in lower amounts in patients with GC. MiR-145-5p was found to affect GC cell proliferation, migration, and invasion by negatively regulating SERPINE1 levels and controlling the ERK1/2 pathway.

Aspleniumguodanum (Aspleniaceae), a distinct new fern species from northern Guangdong, China, based on morphological data and molecular phylogeny.

A new spleenwort species, Aspleniumguodanum, was found and described from Danxia landform region in Guangdong, China. The new species has close resemblance to A.subcrenatum Ching ex S.H.Wu in morphology, but can be distinguished by having plants small, stipes and rachises not covered with fibrous scales, relatively fewer pairs of pinnae, pinnae short, pinna margin weakly biserrate, pinna apex acute and lower pinnae obviously reduced. Phylogenetic analyses, based on six plastid markers (atpB, rbcL, rps4 & rps4-trnS and trnL & trnL-F) of the new species and its relatives, support a close relationship between A.guodanum and A.subcrenatum. Only one population with no more than 50 individuals were found and, therefore, it is recommended to be classified as Critically Endangered (CR) following IUCN Red List Criteria.

Wikstroemiafragrans (Thymelaeaceae, Daphneae), a new species from Mount Danxia, China based on morphological and molecular evidence.

A new species, Wikstroemiafragrans (Thymelaeaceae, Daphneae), from Danxiashan National Park, Shaoguan, Guangdong of China is described and illustrated. It is similar to the sympatric W.trichotoma, but can be differentiated easily from the latter by its shorter racemose inflorescences, yellowish green calyx tube, and smaller leaves. It also resembles the allopatric W.fargesii, but differs from it by its strigose-pubescent ovary and disk scale that is 2- or 3-dentate apically. Phylogenetic analysis using the nuclear DNA internal transcribed spacer (ITS) region revealed that W.fragrans falls within the Wikstroemia clade; based on current sampling, W.fragrans is closely-related to W.capitata. It is also the first species of Wikstroemia known to be endemic to the Danxia landform and is classified provisionally as Critically Endangered according to the IUCN Red List Categories and Criteria.

SH3PXD2A-AS1/miR-330-5p/UBA2 ceRNA network mediates the progression of colorectal cancer through regulating the activity of the Wnt/ß-catenin signaling pathway.

Long non-coding RNAs have important roles in the occurrence and progression of various cancers. However, the molecular mechanism of lncRNAs in colorectal cancer (CRC) is not well illustrated. Thus, we used bioinformatics methods to find potential lncRNAs associated with CRC progression, and chose SH3PXD2A-AS1 as a candidate for further analysis. The roles of SH3PXD2A-AS1 in CRC cells were determined by CCK-8, transwell invasion, wound healing and flow cytometry assays. Besides, we established the CRC tumor models in nude mice to study the effect of SH3PXD2A-AS1 on the tumor growth. Based on the ceRNA hypothesis, we used miRDB and miRTarBase websites to identify the SH3PXD2A-AS1-related ceRNA regulatory network, and measured the roles of this network in CRC cells. The results revealed that the expression profiles of SH3PXD2A-AS1 from GEO and TCGA databases showed an aberrant high level in CRC tissues compared with colorectal normal tissues. SH3PXD2A-AS1 over-expression was also found in CRC cells. SH3PXD2A-AS1 knockdown inhibited the CRC cellular proliferation, invasion and migration but induced apoptosis. Besides, SH3PXD2A-AS1 knockdown also suppressed the growth of CRC tumors. Furthermore, SH3PXD2A-AS1 could function as a ceRNA of miR-330-5p. Additionally, UBA2 was proved to be a target gene of miR-330-5p. Moreover, SH3PXD2A-AS1 knockdown downregulated UBA2 expression through sponging miR-330-5p to inactivate the Wnt/ß-catenin signaling pathway, thereby inhibiting the cell growth and promoting apoptosis. Therefore, the SH3PXD2A-AS1/miR-330-5p/UBA2 network could regulate the progression of CRC through the Wnt/ß-catenin pathway. These findings offer new sights for understanding the pathogenesis of CRC and provide potential biomarkers for CRC treatment.

Transactive response DNA binding protein of 43/histone deacetylase 6 axis alleviates H 2 O 2 -induced retinal ganglion cells injury through inhibiting apoptosis and autophagy.

Oxidative damage is believed to contribute to the pathogenesis of diabetic retinopathy (DR). The current study aimed to detect the effects of transactive response DNA binding protein of 43 (TDP-43) on cell damage induced by hydrogen peroxide (H2 O2 ) in retinal ganglion cells (RGCs) and to investigate the molecular mechanisms involved in this process. We observed that TDP-43 was highly expressed in RGC-5 cells induced by H2 O2 , and that repression of TDP-43 obviously ameliorated H2 O2 -induced RGC-5 cell injury. In addition, loss of TDP-43 profoundly mitigated H2 O2 -triggered oxidative stress by decreasing the production of intracellular reactive oxygen species and the activity of oxidative stress indicator malondialdehyde, as well as enhancing the content of antioxidant enzymes superoxide dismutase, glutathione peroxidase and catalase to restore the antioxidant defense system. Moreover, suppression of TDP-43 obviously obstructed H2 O2 -induced apoptosis. Meanwhile, knockdown of TDP-43 attenuated the expression of the proapoptotic proteins Bax and Cytochrome c, elevated the anti-apoptotic protein Bcl-2, and suppressed the activation of caspase 3 in H2 O2 -induced RGC-5 cells. Moreover, elimination of TDP-43 inhibited H2 O2 -triggered autophagy, which appeared as decreased expression of LC3II/I and Beclin-1, along with p62 degradation. Importantly, silencing of TDP-43 diminished the expression of histone deacetylase 6 (HDAC6), and HDAC6 also abolished the inhibitory effect of TDP-43 inhibition on H2 O2 -induced apoptosis and autophagy. Collectively, our findings demonstrated that depletion of TDP-43 may protect RGC-5 cells against oxidative stress-mediated apoptosis and autophagy by suppressing its target HDAC6. Thus, the TDP-43/HDAC6 axis might be a promising strategy for the treatment of DR.

Simultaneous endoscopic submucosal dissection for multiple early gastric cancers in a low volume center.

The aim of the study is to evaluate the safety and efficacy of simultaneous endoscopic submucosal dissection (ESD) for multiple early gastric cancers.A total of 70 solitary early gastric cancers from 70 patients and 20 multiple early gastric cancers from 10 patients were included in this retrospective study. The curative resection rate, en bloc resection rate, procedure-related complications, and local recurrence were compared between the 2 groups.There was no statistical difference in the rate of complete resection, en bloc resection, and curative resection between the 2 groups (P > .05). No significant difference was found with respect to the occurrence of postoperative bleeding (P > .05). Procedure time was significantly longer in the simultaneous group than that in the single group (87.6 ±â€Š25.1 min vs 54.6 ±â€Š22.0 min, P = .004). The overall incidence of synchronous early gastric cancer was 7.5%.Simultaneous ESD for multiple early gastric cancers is a safe and feasible choice in low-volume hospital. The entire stomach should be examined meticulously during and after ESD. Larger randomized studies are needed to validate our results.

A new azine derivative colorimetric and fluorescent dual-channel probe for cyanide detection.

A novel azine derivative colorimetric and fluorescent dual-channel probe salicylaldehyde hydrazine-3,5-dibromosalicylaldehyde (1) has been designed, synthesized and characterized. The probe 1 is confirmed to have especial selectivity and good sensitivity on detecting CN- via UV-vis absorption and fluorescence spectrum in aqueous solution (H2O/DMSO, 1:4, v/v). This colorimetric and fluorescent dual-channel probe response to CN- owed to the deprotonation process and established the mechanism by using 1H NMR spectroscopy. Further researches showed that the detection limit of the probe 1 to CN- anions is 8.01×10-9M, significantly lower than the maximum level 1.9×10-6M in potable water from WHO guidelines.

Comparison of Endoscopic Resection and Minimally Invasive Esophagectomy in Patients With Early Esophageal Cancer.

OBJECTIVE: To determine whether endoscopic resection (ER) and minimally invasive esophagectomy (MIE) are safe and effective for treating squamous intraepithelial neoplasia of the esophagus. MATERIALS AND METHODS: This study retrospectively analyzed a total of 99 consecutive patients with pathologically confirmed early esophageal cancer between December 2007 and 2011. ER was performed in 59 patients, whereas MIE was performed in 40 patients. We compared the 2 groups according to R0 resection rates, treatment-related complications, mean hospital stay, local recurrence rates, and 3- and 4-year overall survival. RESULTS: No significant differences were found in the R0 resection rates between ER and MIE (94.9% vs. 97.5%, P>0.05). The occurrence rate of minor complications in the ER group was significantly lower than that in the thoracoscopic esophagectomy group (11.8% vs. 32.5%, P>0.05). The mean operative time in the ER group was 74±23 minutes, which was significantly shorter than that in the MIE group (298±46 min). The average length of hospital stay in the ER group was significantly shorter than that in the MIE group (P<0.001). No significant differences were observed in the local recurrence rates between the 2 groups (P>0.05). Similarly, no differences were found in the 3-year survival rate (ER: 96.6%, vs. MIE: 97.5%, P>0.05) and 4-year survival rate (ER: 91.5% vs. MIE: 90%, P>0.05) between the 2 groups. CONCLUSIONS: ER achieves the same positive results as MIE in the treatment of early esophageal cancer and is associated with a lower complication rate, a shorter recovery time, and a similar survival rate. However, multiple ER procedures were required for several patients in this study.

Clinicopathological analysis of endoscopic submucosal dissection for esophageal mucosa lesion / 中国内镜杂志

Objective To analyze the endoscopic and clinicopathologic features of early esopheal carcinoma and precancerous lesions and evaluate the necessity, efficacy and safety of ESD in the treatment. Methods From May 2013 to April 2016, 51 consecutive patients underwent high-resolution video endoscopy and biopsy, confirmed diagnosis of early esophageal squamous cell carcinoma or intraepithelial neoplasia were included. There were capillary loops (IPCL), iodine-staining, preoperative and postoperative pathology, and complications to analyze. Results 51 patients had total 58 lesions, Type A, Type B1, Type B2 of IPCL classification were diagnosed in 8 (13.79%), 44 (75.86%), 6 (10.34%). Low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia, early esophageal carcinoma of preoperative biopsy were diagnosed in 11 (18.97%), 42 (72.41%), 5 (8.62%), low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia, early esophageal carcinoma of postoperative pathology results were diagnosed in 10 (17.54%), 27 (46.55%), 21 (36.21%), concordance rate of pathological results were 60.34%. Complications included micro-perforations (0.00%), strictures (8.62%) and delayed hemorrhage (3.51%), respectively. Conclusion After endoscopic submucosal dissection, detection rate of early esophageal cancer increased significantly, preoperative biopsy had guidance significance in diagnosis and treatment, ESD treatment can reduce the missed diagnosis of early esophageal carcinoma.

Clinicopathological analysis of endoscopic submucosal dissection for esophageal mucosa lesion / 中国内镜杂志

Objective To analyze the endoscopic and clinicopathologic features of early esopheal carcinoma and precancerous lesions and evaluate the necessity, efficacy and safety of ESD in the treatment. Methods From May 2013 to April 2016, 51 consecutive patients underwent high-resolution video endoscopy and biopsy, confirmed diagnosis of early esophageal squamous cell carcinoma or intraepithelial neoplasia were included. There were capillary loops (IPCL), iodine-staining, preoperative and postoperative pathology, and complications to analyze. Results 51 patients had total 58 lesions, Type A, Type B1, Type B2 of IPCL classification were diagnosed in 8 (13.79%), 44 (75.86%), 6 (10.34%). Low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia, early esophageal carcinoma of preoperative biopsy were diagnosed in 11 (18.97%), 42 (72.41%), 5 (8.62%), low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia, early esophageal carcinoma of postoperative pathology results were diagnosed in 10 (17.54%), 27 (46.55%), 21 (36.21%), concordance rate of pathological results were 60.34%. Complications included micro-perforations (0.00%), strictures (8.62%) and delayed hemorrhage (3.51%), respectively. Conclusion After endoscopic submucosal dissection, detection rate of early esophageal cancer increased significantly, preoperative biopsy had guidance significance in diagnosis and treatment, ESD treatment can reduce the missed diagnosis of early esophageal carcinoma.

Detection and Analysis of Ebola Virus in Sierra Leone-China Friendship Biosafety Laboratory from March 11 to April 20, 2015.

Ebola virus disease reemerged in Western Africa in 2014. Chinese Center for Disease Control and Prevention dispatched the first Ebola virus (EBOV) detection team to run newly established Sierra Leone-China Friendship Biological Safety Laboratory. The aims of study were to understand epidemiology, clinical manifestations and survival time of EBOV in patient's blood. A total of 913 specimens were tested between March 11 and April 20, 2015. EBOV positivity occurred in 7.37% of the blood and 0.53% in throat swabs. Most commonly reported symptoms of laboratory confirmed patients were intense fatigue, anorexia, and fever. EBOV RNAs persisted in blood for almost 4 weeks and the real-time RT-PCR Ct values showed close correlation with the sampling time after onset.

Good laboratory practices guarantee biosafety in the Sierra Leone-China friendship biosafety laboratory.

BACKGROUND: The outbreak of Ebola virus disease (EVD) in West Africa between 2014 and 2015 was the largest EDV epidemic since the identification of Ebola virus (EBOV) in 1976, and the countries most strongly affected were Sierra Leone, Guinea, and Liberia. FINDINGS: The Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab), a fixed Biosafety Level 3 laboratory in the capital city of Sierra Leone, was established by the Chinese government and has been active in EBOV detection since 11 March 2015. Complete management and program documents were created for the SLE-CHN Biosafety Lab, and it was divided into four zones (the green, yellow, brown, and red zones) based on the risk assessment. Different types of safe and appropriate personnel protection equipment (PPE) are used in different zones of the laboratory, and it fully meets the Biosafety Level 3 laboratory standards of the World Health Organization. CONCLUSION: Good preparedness, comprehensive risk assessment and operation documents, appropriate PPE, effective monitoring and intensive training, together with well-designed and reasonable laboratory sectioning are essential for guaranteeing biosafety.

Multiband Mucosectomy Versus Endoscopic Submucosal Dissection for Treatment of Squamous Intraepithelial Neoplasia of the Esophagus.

BACKGROUND & AIMS: We compared the efficacy and safety of multiband mucosectomy (MBM) vs endoscopic submucosal dissection (ESD) for the treatment of squamous intraepithelial neoplasia of the esophagus. METHODS: We performed a retrospective study of 78 patients with squamous intraepithelial neoplasia of the esophagus who received either ESD or MBM between January 2009 and January 2011 at the Tengzhou Central People's Hospital in China. We compared rates of bloc resection and curative resection, as well as complications and local recurrence, between groups. RESULTS: Overall, there was no statistical difference in the rate of complete resection between patients who received ESD (95.8%) vs MBM (93%) (P > .05). For tumors less than 15 mm in width, ESD produced a significantly higher rate of en bloc resection (100%) and curative resection (92.3%) than MBM (44.8% and 41%; P < .05). No significant differences were found between lesions less than 15 mm. MBM had a significantly shorter procedure time (38 ± 11 min) than ESD (84 ± 35 min) (P < .05). Major bleeding occurred in 1.85% of MBM procedures and in 16.7% of ESD procedures (P > .05). ESD led to perforations in 8.3% of cases, whereas MBM did not lead to any perforations (P < .05). No significant differences were found between groups in proportions of cases with postoperative esophageal strictures (16.7% vs 14.8%; P > .05) or the 3-year rate of local recurrence (P > .05). CONCLUSIONS: Based on a retrospective comparison of patients who underwent ESD vs MBM for squamous intraepithelial neoplasia of the esophagus, ESD should be reserved for patients with larger neoplastic lesions (>15 mm), with respect to the success of attempted en bloc resection and the number of curative resections achieved. However, ESD has longer procedure times and higher rates of complication. MBM allows for safe and easy piecemeal resections, and is associated with similar levels of clinical success as ESD for lesions less than 15 mm. Large, randomized, controlled studies are needed to determine which endoscopic resection modality is superior for patients with high-grade intraepithelia neoplasms.

[Expression of neuraminidase gene of influenza virus H1N1 in baculovirus-expression system].

OBJECTIVE: To construct the recombinant baculovirus with NA gene of Influenza H1N1 virus. METHODS: Full-length NA gene of Influenza virus H1N1 (A/PR/8/34) was amplified by PCR and inserted into pFastBacdual vector to construct the recombinant baculovirus transfer vector pFBD-NA. Recombinant shuttle vectors rBacmid-NA was then obtained after transforming DH10B competent cells containing bacmid plasmids. After transfecting into sf9 cells, recombinant baculovirus rBac-NA was obtained. The rBac-NA genome was extracted and identified by PCR. NA protein expressed by recombinant baculovirus-infected sf9 cells was determined by IFA, Western Bolt and ELISA. RESULTS: PCR results proved that recombinant shuttle vectors rBacmid-NA was successfully constructed. NA protein was detected by IFA and showed strong specific green fluorescence on the surface of infected cells. NA protein was recognized by two polyclonal antibodies specific for NA in Western Blot. ELISA showed specific reaction of recombinant NA protein with mouse polyclonal antibody against influenza virus (PR8), indicating high antigenicity. CONCLUSION: Recombinant baculovirus rBac-NA that expresse NA protein of influenza virus was successfully constructed. This work provides a basis for further study on NA protein function and novel influenza vaccine development.

Catestatin could ameliorate proliferating changes of target organs in spontaneously hypertensive rats.

BACKGROUND: Catestatin, a chromogranin A-derived peptide, is a potent antagonist of nicotine-evoked catecholamine release. We know that catecholamine plays an important role in cardiovascular remodeling induced by hypertension, therefore we hypothesized that catestatin would affect target-organ structure during hypertension. METHODS: Twelve spontaneously hypertensive rats (SHRs) were randomized to SHR control group and catestatin group, the normal control group was comprised of six healthy Wistar-Kyoto rats of the same age. Tail-cuff blood pressure and pulse rate were obtained at weeks 1, 4 and 8. At the end of the eight-week period, the heart, abdominal aorta and left kidney were excised and weighed, VG staining was done and the intima-media thickness of vessels and the collagen volume fraction were assessed by an image acquisition and analysis system. The proliferating cell nuclear antigen (PCNA) was observed by immunohistochemistry, and real time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of proliferative genes including cyclin A, ki67 and PCNA in the abdominal aorta. RESULTS: All the parameters in SHR observed in the present study increased significantly compared to Wistar Kyoto rats (P < 0.01). With intervention with catestatin, the systolic blood pressure decreased slightly but it was not significantly different from the SHR control, the cardiac mass index and left ventricular mass index both decreased significant ly, the collagen volume fraction decreased by nearly 30% in the heart, by 25% in vessels and by 10% in the kidney, and the intima-media thickness and expression of proliferative genes, including cyclin A, ki67 and PCNA, in the abdominal aorta also decreased significant ly. CONCLUSIONS: The present study indicated that catestatin could ameliorate proliferating changes of heart, kidney and vessels during hypertension, especially to the deposition of interstitial collagen. Blood pressure was not the main factor to mediate this effect, which suggested that catestatin could become a novel protective factor for hypertensive target organs.

Effect of aliskiren on arterial stiffness, compared with ramipril in patients with mild to moderate essential hypertension.

BACKGROUND: Aliskiren is a novel blood pressure-lowering agent acting as an oral direct renin inhibitor. The aim of this study was to assess the effect of aliskiren on arterial stiffness, compared with that of ramipril in mild to moderate essential hypertensive patients. METHODS: Following a two week placebo run-in period, patients with a mean sitting diastolic blood pressure (ms-DBP) ≥ 95 and < 110 mmHg (1 mmHg = 0.133 kPa), and a mean sitting systolic blood pressure (ms-SBP) < 180 mmHg were randomly allocated to treatment with aliskiren (150 mg/d, n = 20) or ramipril (5 mg/d, n = 20) for eight weeks. Blood pressure, plasma renin activity, and the brachial-ankle pulse wave velocity (ba-PWV) were measured before and after eight weeks of treatment. RESULTS: Eight weeks of treatment significantly decreased systolic blood pressure and diastolic blood pressure in both the aliskiren group and ramipril group. The hypotensive effect did not differ between the two groups. Plasma renin activity decreased after aliskiren treatment and increased after ramipril treatment. There was no significant difference in baseline ba-PWV between the aliskiren and ramipril groups (P = 0.892). The ba-PWV was significantly reduced in both the aliskiren group (1535 (1405 - 1666) vs. 1464 (1360 - 1506) cm/s) (P < 0.01) and the ramipril group (1544 (1433 - 1673) vs. 1447 (1327 - 1549) cm/s) (P < 0.01). No statistically significant difference was found in the decline of ba-PWV between the two groups (P = 0.766). CONCLUSIONS: The current study revealed that aliskiren (150 mg/d) could ameliorate arterial stiffness and its effect was similar to ramipril (5 mg/d) in mild to moderate hypertensive patients, indicating that in addition to lowering blood pressure, aliskiren had beneficial effect on vascular protection.

[Establishment of mammalian cell lines for constitutive expression of influenza virus matrix protein 2].

To establish a mammalian cell line for stable expression of the matrix protein 2 (M2) of influenza virus type A. M2 gene was amplified by PCR from the influenza virus strain A/PR/8/34. The PCR product was cloned into eukaryotic expression vector pcDNA5/FRT. After identification with restriction enzyme digestion, the plasmid was co-transfected with plasmid pOG44 which expressed Flp in Flp-In-CHO cells. The target gene was integrated into chromosome of CHO cells by homologous recombination in vivo. Recombinant CHO-M2 cell lines were selected for hygromycin B resistance. A total of 15 recombinant cell strains with high expression of M2 protein were screened by hygromycin, and the expression of M2 protein was determined by IFA and Western blot. After subculturing for 10 passages, the presence of M2 gene in the CHO-M2 cells was confirmed by PCR, and the expression of M2 protein were proved by IFA and Western blot. We successfully constructed a mammalian cell line which stably expressed M2 protein of influenza virus type A. The cell line will be useful for studies on function of M2 protein and provide tools for novel influenza virus vaccine development.

In-vivo investigation of laser-induced choroidal neovascularization in rat using spectral-domain optical coherence tomography (SD-OCT).

PURPOSE: This study investigated the in-vivo formation process of laser-induced choroidal neovascularization (CNV) in rat using high-resolution spectral-domain optical coherence tomography (SD-OCT), and compared the results to histological methods. METHODS: Brown Norway rats (n = 60, 6-8 weeks of age) received 532-nm diode laser photocoagulation. SD-OCT and fluorescein angiography (FA) were performed in vivo 2, 5, 7, 14, and 21 days post-laser application. Haematoxylin and eosin (H&E) staining and immunohistochemistry for CD31, phosphorylated vascular endothelial factor receptor 2 (pVEGFR2) were conducted at each time point to observe the CNV in vitro. Choroidal flatmount preparations were observed using a confocal laser scanning microscope (CLSM) and a scanning electron microscope (SEM). RESULTS: SD-OCT monitored the longitudinal morphological changes of laser-induced CNV. CNV reached its maximal size on day 7, and began a gradual reduction on day 14. FA revealed similar dynamic changes in leakage. CNV thickness, as assessed by SD-OCT, was consistent with H&E-stained sections at each time point. CLSM and SEM revealed the details of the fibrovascular membrane. CD31 and pVEGFR2 expression supported the results of SD-OCT and histology. CONCLUSIONS: SD-OCT was a convenient and reliable tool for the imaging of the CNV formation process and quantification of the lesion size in vivo.

[Construction of recombinant baculovirus co-expressing M1 and HA of influenza A virus].

The M1 and HA genes of H1N1 influenza virus were amplified and then cloned into the pFastBac dual donor plasmid. The recombinant pFastBac Dual-M1-HA was identified by restriction enzyme digestion. After the pFastBacdual-M1-HA was transformed into the baculovirus shuttle plasmid (bacmid) in DH10Bac competent cells, the colonies were identified by antibiotics and blue-white selection. The rBac-mid-M1-HA was verified by PCR and transfected into S f9 cells to produce recombinant baculovirus (rBac-M1-HA). Gene insertion of rBac-M1-HA was verified and the expression of M1 and HA genes was analyzed by IFA and Western-blot, demonstrating M1 and HA were co-expressed successfully. This study provides the foundation for researching the formation mechanism of influenza VLP and developing new influenza vaccines.

Different expression patterns of CEACAM1 and its impacts on angiogenesis in gastric nonneoplastic and neoplastic lesions.

OBJECTIVE: This study was designed to investigate the expression patterns of CEACAM1 and its relationship with angiogenesis in nonneoplastic and neoplastic gastric lesions. METHODS: CEACAM1 and TGF-ß expression was detected by immunohistochemical staining and dual-labeling immunohistochemical staining in neoplastic and nonneoplastic lesions. MVD-CD31 and MVD-CD105 were counted in CEACAM1-positive areas by dual-labeling immunohistochemistry. RESULTS: There was no expression of CEACAM1 in normal gastric mucosa. In IM and GIN, CEACAM1 was mainly expressed with membranous pattern. CEACAM1 was expressed with membranous pattern in well-differentiated adenocarcinoma, with cytoplasmic pattern in poorly differentiated adenocarcinoma, and with cytoplasmic and membranous pattern mixed together in intermediately adenocarcinoma. The expression patterns of CEACAM1 showed a significant difference (P < 0.05) in nonneoplastic and neoplastic lesions. Coexpression of CEACAM1 and TGF-ß was elevated and significantly different from nonneoplastic to neoplastic lesions (P < 0.05). Moreover, CEACAM1 and TGF-ß coexpression were related to carcinoma progression (r = 0.35; P < 0.05). MVD-CD31 and MVD-CD105 showed significant differences from nonneoplastic to neoplastic lesions (P < 0.05). CONCLUSIONS: CEACAM1 has different expression patterns in nonneoplastic and neoplastic lesions. The coexpression of CEACAM1 and TGF-ß increased from nonneoplastic to neoplastic lesions and may be related with tumor progression via promoting tumorous angiogenesis.