Objective Study the relationship between type A influenza virus genetic variation with survival selective pressure, help for the finding of possible vaccine conserved antigen target. Methods Select seven strains of same HA (Hemagglutinin) serotype, regional and isolation time closely related type A influenza virus with full HA gene coding sequence; use Blast2 program to calculate the parameter of nucleotide conservative, amino acid conservative, mutation ratio of codon 3rd (over) non 3rd locus, survival selective pressure indicator of these virus strains; analysis the parameters relationship with survival selective pressure indicator. Results Nucleotide conservative of HA gene is significantly lower than that of other genes; amino acid conservative of HA gene is similar with NS gene, all lower than that of other genes; genetic variation of HA gene is under survival selective pressure; selective pressure toward NS, PB1 gene is relatively lower than that toward NA, NP gene; survival selective pressure indicator is strongly correlated with mutation frequency upon codon 3rd locus. Conclusions Genetic variation of influenza virus is determined both by mechanism of relax replication model and survival selective pressure; genetic conservative of each gene is different, NA, NP gene could be selected as possible vaccine target for their relative high conservative, HA gene possesses medium genetic conservative with prospective of finding more conserved epitope region within its full sequence, NS, PB1 are not recommended as vaccine candidate for their relative low genetic conservative.

ObjectiveTo establish and evaluate the methods which can quantify the ocular hemodynamics by combined scanning laser ophthalmoscopy video fluorescein angiography with computerized digital image processing system．MethodsAfter acquiring serial digital images by video capture from some angiography videotapes and measuring the gray values of some fixed areas over time in these serial images，the gray value curves and some parameters were obtained．The inter- and intra-observer reproducibility surveys were carried out．ResultsThe hemodynamic parameters were obtained，which included sectional choroidal filling time and rate，sectional papillary filling time and rate，retinal arterial and venous filling time and rate，arterovenous passing time and diameter of artery and vein．The intra- and inter-observer reproducibility were fairly good．Conclusioncomputerized digital image processing system is useful for understanding retinal and choroidal circulation．

Background Anti-scarring is a focus following glaucoma filtering surgery.It has been verified that expression of heat shock protein 47 (HSP47) is always correlated with that of various types of collagens and closely related with the collagen-related diseases including fibrosis in various organs.However,the relationship between the expression of HSP47 and fibrosis of filtering bleb is unclear.Ohjective Present study was to investigate the dynamic expression of HSP47 in subconjunctival filtering bleb after anti-glaucomatous filtering surgery.Methods Filtering surgery was performed on the right eyes of 10 SPF male Spargue-Dawley(SD) rats.Then the operative eyes were randomized into the postoperative 7-day group and 14-day group according to the randomized number table,and the left eyes of the rats served as the normal control group.The shape of filtering blebs and the ocular anterior segment response were examined under microscope daily.The animals were sacrificed by excessive anesthesia on day 7 and 14 after operation,respectively,and the specimens were prepared of the eyes.Regular histopathological examination was performed to observe the collagen fibrosis change,and immunohistochemistry was carried out to assay the dynamic expression of HSP47 depended on the fibrosis.Percentage of positive cells for HSP47 were calculated.The use and care of the experimental animals complied with the Regulation for the Administration of Affair Concerning the Experimental Animals by State Science and Technology Committee.Results Diffuse subconjunctival blebs were seen postoperatively in t0 eyes after surgery with mild inflammatory response in ocular anterior segment.Hematoxylin-eosin staining showed that compared with the specimens of the normal control group,fibroblasts obviously proliferated and collagenous fibers increased in the specimens of operative group.The HSP47 protein was less expressed in cytoplasm of many fibroblasts and less vascular endothelial cells in the normal rat specimens.The expressing intensity of HSP47 protein was enhanced.However,the expression intensity was weaker in the postoperative 14-day group in comparison with the 7-day group.The percentage of positive cells for HSP47 in the filtering blebs 7 days after operation was (56.40±1.12)％,that in the normal control group was (13.70±0.74)％,with a significant difference between them (P=0.000).In the postoperative 14-day group,the percentage of positive cells for HSP47 reduced from (56.40± 1.12) ％ to (23.90±0.76) ％ (P =0.000),but it was still higher than that of the normal control group (P =0.000).Conclusions Expression intensity of HSP47 alters as the activity of fibroblasts in filtering bleb,suggesting that HSP47 plays a role in collagen-fibrosis following filtering surgery in SD rats.

Objective To explore the clinical application value of serum amyloid protein(SAA) and C reactive protein(CRP)in early diagnosis of hand‐foot‐mouth disease(HFMD)in children .Methods The serum/plasma levels of CRP and SAA in 81 children with HFMD(HFMD group)and 125 healthy children(healthy control group)were detected by using dot immunogold filtration as‐say .Data including white blood cell (WBC) count ,levels of CRP and SAA were obtained and analyzed .And diagnostic efficacies of these indicators were evaluated by using receiver operating characteristic (ROC) curve .Results The serum levels of SAA and CRP and WBC count in HFMD group were significantly higher than those in healthy control group ,and all had statistically significant differences(P<0 .05) .The area under the ROC curve(AUC) of SAA(0 .921) was higher than that of CRP(0 .728) ,SAA/CRP ra‐tio(0 .846) and WBC count(0 .721) for diagnosing HFMD ,there were statistically significant differences(P<0 .05) .Compared with CRP ,WBC count and SAA/CRP ratio ,SAA had better diagnostic efficacy .Conclusion As a new biomarker of infections ,SAA can effectively diagnose HFMD early and provide more sensitive reference to clinicians for early diagnosis ,which might be worthy of clinical application widely .

The present paper introduces the definition of prehypertension,summarizes the advances in prehypertensive epidemiology and focus of debate and suggests the outlook of prehypertension.

Objective To verify the application of anterior segment perfusion culture and trabecular meshwork (TM) organ culture for glaucoma study. Methods TM tissue was cultured by perfused anterior segment and TM organ culture, light microscopy was used to observe the TM cells and intercellular spaces.Results IOP of the porcine anterior segments perfused under constant flow at 0.1 mL/h could bekept in normal range(10～12 mmHg). The IOP was elevated with the increasing of perfusion rate, while the morphology and structure of the tissue were well preserved. TM cultured by TM organ culture could also reserve the tissues well, but the intercelluar spaces collapsed. Conclusion Anterior segment perfusion model could be a short-term high-pressure model and may simulate the normal physical state. Adequate perfusion was necessary for normal TM.

Chinese traditional culture is the soil for the survival and development of traditional Chinese medi-cine,among them,the concept of A thing is valued if it is rare portal factions and master has influenced the development,communication and popularization of traditional Chinese medicine.But the spirit and thinking of harmony unity of heaven and man comparative state benevolence and self-cultivation and self-re-straint has played an active role in the development of traditional Chinese medicine.Thus this paper put forward the suggestions and opinions of strengthening the cultivation of humanistic spirit of higher traditional Chinese medi-cine talents:establishing a comprehensive evaluation standards of higher traditional Chinese medicine talents,es-tablishing scientific development concept to treat the inheritance and innovation of traditional Chinese medicine cul-ture,perfecting the curriculum provision of relevant professions,creating the conducive environment for the trans-mission of traditional Chinese medicine culture,hoping to provide a reference for the explanation of the relationship between traditional culture and traditional Chinese medicine.

OBJECTIVE: To investigate the postoperative application of nasopharyngeal airway (NPA) in rhinogenous obstructive sleep apnea hypopnea syndrome (OSAHS) patients, so that to observe the parameters including vital signs of the patients and evaluatethe value of clinical application and reliability of NPA. METHOD: A total of 216 patients diagnosed as rhinogenous OSAHS were randomly assigned to experimental group (setting NPA, 112 cases) and control group (not setting NPA, 104 cases) according to whether NPA was placed in the nasal cavity postoperatively. ECG, oxygen saturation and hemodynamics were monitored for 24 h postoperatively. The pharyngeal pain and discomfort, low oxygen saturation and hemodynamics were compared between these two groups. The subjective assessment and clinical symptoms were compared between the two groups using visual analogue scale (VAS). RESULT: The experimental group showed better relief of nasal obstruction, nasal pain, headache, dry pharynx, insomnia and pain while taking out nasal packing compared with control group, where the differences were statistically significant (P < 0.01). In the experimental group, the level of LSa2O2 (P < 0.05), HR (P < 0.01), SBP (P < 0.05), DBP (P < 0.01), MAP (P < 0.01) and RPP (P < 0.01) was significantly lower than in the control group. CONCLUSION The postoperative application of nasopharyngeal airway in rhinogenous OSAHS patients could help to keep nasal patency and avoid the upper airway obstruction, which exhibited good safety and compliance. The nasopharyngeal airway can reduce patients' discomfort and improve hyoxemia, ensuring hemodynamic stability.
Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nasopharynx ; Postoperative Care ; Respiration, Artificial ; methods ; Sleep Apnea, Obstructive ; therapy ; Young Adult

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.