Background Femtosecond laser in situ keratomileusis (LASIK) inevitably injury keratocytes and corneal nerve fibers.The research report about postoperative morphological changes of corneal nerve regeneration and keratocytes in femtosecond LASIK is still rare.Objective The aim of this study was to observe the kinetic changes of keratocytes and corneal nerve in corneal flap after femtosecond LASIK.Method Femtosecond laser manufacture corneal flap of LASIK surgery was performed on 60 eyes of 30 patients with refractive error using both femtosecond laser system and excimer laser treatment system.The repair of corneal wound was examined by slit lamp microscope,and the morphology of keratocytes and corneal nerve were observed with confocal microscope 1 week,1month,3 months after surgery,respectively.Results No haze or flap folds were found under the slit lamp microscope from 1 week through 3 months after operation.One week after surgery,the corneal stromal cells at the interface of the corneal flap appeared to be a mild activation status in 42 eyes (70％),but the activated cells gradually reduced with lapse of time.Three months after surgery,mild activation state still was found in 7 eyes (12％).One week after surgery,independent,short (＜50 μm),curved subbasal nerve fibers were exhibited in 7 eyes (12％),and curved filamentous nerve fibers were discovered in 48 eyes (80％) one month after surgery.The nerve fiber length of subbasal nerve was ＞200 μm in 27 eyes (45％) and classes beaded structure appeared 3 months after operation but were still different with preoperative subbasal nerve fibers.One week after the operation,filaments or discontinuous nerve fibers could been seen in 46 eyes (77％) at theinterface,and long nerve fibers or filamentous nerves were visible around the terminal or periphery of nerve fibers in 49 eyes (82％) one month after surgery,and long nerve fibers or filaments of nerve fibers were visible in 57 eyes (95％) 3 months after the surgery.Conclusions Femtosecond LASIK cause wound reaction at cellular level.Corneal nerve fibers recover with the extension of time,but there are still some morphological differences 3 months after surgery from preoperation.

In vivo fluorescence molecular imaging plays a more and more important role in the observation of diseases, drug research and biology research because of its low cost, simplicity and no ionizing radiation to biological tissue. Herein, the most important parts of the optical fluorescence molecular imaging and their advances are summarized, including fluorescent molecular probes, imaging systems and reconstruction algorithms. The application and development trend of this technology are also introduced in this paper.
Algorithms ; Fluorescent Dyes ; Molecular Imaging ; methods

It is believed that in the RNA world the operational (ribozymes) and the informational (riboscripts) RNA molecules were created with only three (adenosine, uridine, and guanosine) and two (adenosine and uridine) nucleosides, respectively, so that the genetic code started uncomplicated. Ribozymes subsequently evolved to be able to cut and paste themselves and riboscripts were acceptive to rigorous editing (adenosine to inosine); the intensive diversification of RNA molecules shaped novel cellular machineries that are capable of polymerizing amino acids-a new type of cellular building materials for life. Initially, the genetic code, encoding seven amino acids, was created only to distinguish purine and pyrimidine; it was later expanded in a stepwise way to encode 12, 15, and 20 amino acids through the relief of guanine from its roles as operational signals and through the recruitment of cytosine. Therefore, the maturation of the genetic code also coincided with (1) the departure of aminoacyl-tRNA synthetases (AARSs) from the primordial translation machinery, (2) the replacement of informational RNA by DNA, and (3) the co-evolution of AARSs and their cognate tRNAs. This model predicts gradual replacements of RNA-made molecular mechanisms, cellular processes by proteins, and informational exploitation by DNA.
Amino Acid Sequence ; Amino Acyl-tRNA Synthetases ; genetics ; metabolism ; Base Composition ; DNA ; chemistry ; genetics ; metabolism ; Eosinophil Cationic Protein ; chemistry ; genetics ; Evolution, Molecular ; Genetic Code ; Models, Genetic ; Molecular Sequence Data ; RNA ; chemistry ; genetics ; metabolism ; Sequence Homology, Amino Acid

Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.
Animals ; China ; Cloning, Molecular ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Penaeidae ; virology ; Phylogeny ; White spot syndrome virus 1 ; genetics

OBJECTIVETo investigate the effect of Spearmint oil on inflammation, oxidative alteration and Nrf2 expression in rats with chronic obstructive pulmonary disease(COPD).

METHODSCOPD model was induced by intratracheal instillation of Klebsiella pneumonia and lipopolysaccharide (LPS) for 12 weeks in rats, and COPD rats were treated with Spearmint oil for 3 weeks. After COPD was induced, the pathological changes, changes in leucocyte number in blood and bronchoalveolar lavage fluid (BALF), MDA in lung homogenate and Nrf2 expression were observed. The effects of Spearmint oil on these changes were determined.

RESULTSpearmint oil 100 mg*kg(-1)significantly reduced leucocyte numbers in BALF, and attenuated bronchiolitis, pulmonary interstitial inflammation and inflammation cell infiltration. Spearmint oil 30-300 mg*kg(-1)decreased the destruction of pulmonary alveolus and the thickness of bronchioles walls, and inhibited goblet cell proliferation. Spearmint oil significantly reduced MDA in lung homogenate, and decreased the expression of Nrf2 protein in lung tissues.

CONCLUSIONSpearmint oil has protective effect on lung injury in COPD rats, since it improves pulmonary inflammation,oxidative alteration, and enhances Nrf2 protein expression.

Animals ; Klebsiella pneumoniae ; Lipopolysaccharides ; Male ; Mentha spicata ; chemistry ; NF-E2-Related Factor 2 ; metabolism ; Oils, Volatile ; pharmacology ; therapeutic use ; Oxidative Stress ; drug effects ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; etiology ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the biomarkers of early diagnosis in patients with polycyclic aromatic hydrocarbons (PAHs)-related lung cancer for the application to detection of occupational lung cancer or related lung cancer.

METHODSWestern dot blotting was used to explore the expression of ras, p53 and heat stress protein 70 (HSP70) in 29 patients with PAHs-related lung cancer (LC), and 28 patients with non-cancerous pulmonary disease, and 30 healthy controls.

RESULTSThe positive detection rates of P21, P53, and HSP70 in LC group (58.62%, 34.48%, 41.38% respectively) were higher than those in non-cancerous pulmonary disease group (14.29%, 7.14%, 10.71% respectively, P < 0.01). The sensitivity of P21, P53 and HSP70 were 58.62%, 34.48% and 41.38% respectively, negative predictive value (NPV) were 68.42%, 78.05% and 63.04% respectively. The co-detection of the three proteins mentioned above produced a sensitivity of 82.76% with a NPV of 78.26% (P < 0.05). Of 18 cases of LC with negative cytology, 13 (72.22%) were found HSP21, P53 or HSP70 positive.

CONCLUSIONSCo-detection of the P21, P53, and HSP70 may be used as the screening marker for diagnosis of PAHs-related lung cancer, and may supplement the diagnostic value of conventional cytology.

Aged ; Biomarkers ; analysis ; Blotting, Western ; Case-Control Studies ; HSP70 Heat-Shock Proteins ; analysis ; Humans ; Lung Neoplasms ; chemically induced ; metabolism ; pathology ; Middle Aged ; Occupational Exposure ; Polycyclic Aromatic Hydrocarbons ; poisoning ; Proto-Oncogene Proteins p21(ras) ; analysis ; Tumor Suppressor Protein p53 ; analysis

OBJECTIVETo explore the therapeutic effect of interleukin-11 (IL-11) on high-dose methotrexate (HDMTX) induced mucositis in Wistar's rats, the proliferative effect on CEM leukemia cell line and the antitumor effect on HDMTX.

METHODSNinety-five 5-week old, 120 - 150 grams weight Wistar rats were randomly divided into five groups. Group A is normal control (n = 15), group B MTX control (n = 20), group C IL-11 pretreatment group before MTX injection (n = 20), group D (n = 20) the high dose IL-11 group (475 microg.kg(-1).d(-1)) after MTX injection, group E (n = 20) the low dose IL-11 group (150 microg.kg(-1).d(-1)) after MTX injection. All rats in group B approximately E were given 1 ml MTX intraperitoneally (100 mg/kg). Rats were killed at day 1, 3, 5, 7 after MTX injection. The mortality rates, changes of small intestine tissue morphology and ultra structure were observed. The proliferation of small intestine crypt cell was assayed by proliferating cell nuclear antigen (PCNA) immunohistochemical staining. MTT method was used to detect the proliferation of CEM cell line.

RESULTIL-11 treatment resulted in a significant increase of survival of HDMTX treated rats, increased of small intestinal villus length and villus/crypt ratio. IL-11 administration was associated with enhancement of small intestine mucosa recovery after HDMTX therapy. Group C showed a greater effect than group B (P < 0.01). IL-11 had no effect on CEM cell proliferation.

CONCLUSIONIL-11 has a significant mitigating effect on high-dose MTX induced intestinal mucositis in rat, and significantly increase the survival of the rats. IL-11 could be safely used in the HDMTX treatment of childhood acute lymphocyte leukemia.

Animals ; Antimetabolites, Antineoplastic ; toxicity ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Female ; Humans ; Immunohistochemistry ; Interleukin-11 ; pharmacology ; therapeutic use ; Intestinal Mucosa ; drug effects ; pathology ; ultrastructure ; Intestine, Small ; drug effects ; metabolism ; pathology ; Male ; Methotrexate ; toxicity ; Microscopy, Electron ; Mucositis ; chemically induced ; mortality ; prevention & control ; Proliferating Cell Nuclear Antigen ; analysis ; Random Allocation ; Rats ; Rats, Wistar ; Survival Rate

The prognostic value of phosphatidylinositol-4,5-bisphosphate 3-kinase,catalytic subunit alpha (PIK3CA) in patients with esophageal squamous cell carcinoma (ESCC) is controversial.We aimed to investigate the prognostic significance of PIK3CA mutation in patients with ESCC.EMBASE,PubMed,and Web of Science databases were systematically searched from inception through Oct.3,2016.The hazard ratios (HRs) and 95％ confidence intervals (CI) were calculated using a random effects model for overall survival (OS) and disease-free survival (DFS).Seven studies enrolling 1505 patients were eligible for inclusion of the current meta-analysis.Results revealed that PIK3CA mutation was not significantly associated with OS (HR:0.90,95％ CI:0.63-1.30,P=0.591),with a significant heterogeneity (I2=65.7％,P=0.012).Additionally,subgroup analyses were further conducted according to various variables,such as types of specimen,the sample size,technique and statistical methodology.All results suggested that no significant relationship was found between PIK3CA mutation and OS in patients with ESCC.For DFS,there was no significant association between PIK3CA mutation and DFS in patients with ESCC (HR:1.00,95％ CI=0.47-2.11,P=0.993,I2=73.7％).Publication bias was not present and the results of sensitivity analysis were very stable in the current meta-analysis.Our findings suggest that PIK3CA mutation has no significant effects on OS and DFS in ESCC patients.More well-designed prospective studies with better methodology for PIK3CA assessment are required to clarify the prognostic significance of PIK3CA mutation in ESCC patients.

Serum levels of soluble MHC class I-related chain A (sMICA) are related with the prognosis of various types of cancer; however, few studies on the prognostic value of sMICA in hepatocellular carcinoma (HCC) have been reported. In this study, we retrospectively investigated the relationship between sMICA levels and clinical features of advanced HCC, and we assessed the prognostic value of sMICA in advanced HCC. Furthermore, the relationship of serum sMICA levels and natural killer group 2, member D (NKG2D) expression on natural killer (NK) cells was also evaluated. We detected sMICA levels in the serum of 60 advanced HCC patients using enzyme-linked immunosorbent assay (ELISA) and measured expression levels of NKG2D on NK cells using flow cytometry. We found that serum sMICA levels in HCC patients were in the range of 0.10-6.21 ng/mL. Chi-square analyses showed that sMICA level was significantly related with only tumor size. Survival analysis showed that a high sMICA level was significantly related with poor prognosis among HCC patients. Multivariate analyses indicated that sMICA was an independent prognostic factor. In addition, the levels of CD56+NKG2D+ NK cells were within the range of 11.2%-55.4%, and correlation analyses indicated that sMICA level was negatively correlated with the level of NKG2D+ NK cells. Our results suggest that serum sMICA levels may be an independent prognostic factor for advanced HCC.
Adult ; Carcinoma, Hepatocellular ; blood ; immunology ; pathology ; Female ; Histocompatibility Antigens Class I ; blood ; Humans ; Killer Cells, Natural ; immunology ; metabolism ; Liver Neoplasms ; blood ; immunology ; pathology ; Male ; Middle Aged ; Multivariate Analysis ; NK Cell Lectin-Like Receptor Subfamily K ; metabolism ; Neoplasm Staging ; Retrospective Studies ; Survival Rate ; Tumor Burden

Objective To explore the mechanisms of different cholic acid for reducing damage to human liver cells lines L-O2 induced by amanita toxic peptides (amataxins).Methods According to different concentrations of amataxins,the experiment was conducted with different dosages in 5 groups:0.00 g/L,0.26 g/L,0.40 g/L,1.40 g/L and 2.80 g/L.The human liver cells lines L-O2 in the exponential growth phase were cultured into 96-well plates,1 ×103 cells per well After 24 hours,the concentrations of amanita toxic peptides mentioned above were added.The minimum concentration of mushroom toxins keeping the liver cells alive was determined after 24,48 and 72 hours,respectively,and MTT method was used to test each group's liver cell activity.The experiment included 3 groups:the control group,the damage group,and the cholic acid group.Each group had 3 time points:24,48 and 72 hours after being attacked.Twenty four hours after attack,in cholic acid group,cholic acid drugs including taurocholic acid gca,goose deoxycholic acid,gansu ammonia goose deoxycholic acid and bovine goose deoxycholic acid were given,respectively,to protect the injured liver cells.Cells' morphology changes were observed under the inverted phase contrast microscope,living cells were counted by using MTT method,and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the culture supernatant were tested by the biochemical method.Results The minimum attack concentration of lamanita toxic peptides keeping liver cell survival in vitro was 1.40 g/L.Seventy-two hours after attack by amanita toxic peptides,the absorbance value was 0.812 ± 0.035,0.345 ± 0.021,0.363 ± 0.018,0.387 ± 0.027,0.431 ± 0.018,0.465 ± 0.015 and 0.452 ± 0.030,respectively in the control group,the damage group,the taurocholic acid group,the goose deoxycholic acid group,the glycocholic acid group,the glycochenodeoxycholic acid group and the sodium deoxycholic acid group.Compared with the damage group,absorbance value 72 hours after attack in each cholic acid group gradually increased,and compared with damage group,the differences were statistically significant among goose deoxycholic acid group,glycocholic acid group,glycochenodeoxycholic acid group and sodium deoxycholic acid group(P ＜ 0.05).AST and ALT activity in each cholic acid group declined,and that in glycochenodeoxycholic acid group was the lowest.Compared with the damage group,the difference was statistically significant (P ＜ 0.01).Conclusions Cholic acid can protect human liver cells from the damage induced by amanita toxic peptides.Such effect may be related to the fact that both amanita toxic peptides and cholic acid are the substrates of NTCP and OATP.