Ocular surface tissues are vulnerable to various stimuli from the external environment,including microbial invasion,mechanical damage,chemical stimulation,dust,UV light,and allergen stimulation.Conjunctival goblet cells play an important role in maintaining ocular surface self-homeostasis and health.These cells are the only unicellular glands in humans and mammals that are mainly distributed in the epithelium of the digestive tract,respiratory tract,and the ocular surface.Many factors affect goblet cell development and differentiation,especially the newly discovered molecule Spdef,which is a transcription factor found in the ETS family.The main function of conjunctival goblet cells is to secrete mucin to protect and lubricate the ocular surface;however,they also secrete a variety of other bioactive substances.Recent studies have demonstrated that goblet cells are involved in antigen presentation,regulation of dendritic cell phenotype differentiation,and induction of immune tolerance on the mucosa.Many ocular surface diseases,such as dry eye,will alter density and function of conjunctival goblet cells.Therefore,ophthalmologists should be concerned about the biological behavior of conjunctival goblet cells and the relationship between them and ocular surface diseases.

Purpose：To study the incidence of pneumonia, pathogenic bacteria and prognosis in patients with neoplasm after chemotherapy.Methods：Sputum culture was made by routine method. Then,to determine the kinds of bacteria bioMrieux ATB expression was used, and general drug-sensitivity test was done by Kirby-Bautr method.Results：Fever occured in 27.3% patients after chemotherapy, while pneumonia in 11.3% patients. 13 kinds of bacteria, 72 strains of bacteria and 2 kinds of fungi were found in 68 positive culture samples. The infection rates were 76.5% in infection of single bacteoia，23.5% in infection of two kinds of bacteria and 22.1% in bacteria with fungi. The infection rate of gram-negative bacteria is much higher than that of gram-positive ones (P＜0.01).Conclusions：The incidence of pneumonia in the hospital is higher in patients with neoplasm after chemotherapy. Most pathogenic bacteria are gram-negative and sensitive to aminoglycosides, piperacillin and norfloxacin.

Endothelial progenitor cells play a significant role in neovascularization of ischemic tissues and in reendothelialization of damaged blood vessels. Cytokines, an important components of micro-environment of angiogenesis,play an important role in regulation of endothelial progenitor cells. The effective application of cytokines can significantly argument the number, enhance the biological function and improve the therapeutic effect of endothelial progenitor cells, which play a positive role in regulation of endothelial progenitor cells.This article briefly reviewed the regulating mechanisms of the vascular endothelial growth factor and other cytokines in a total of 7 endothelial progenitor cells.

Objective To study the accuracy of an ultrasonographic characteristic diagnostic score system(UCDSS) which was used to classify the breast solid nodular lesions. Methods UCDSS were established by analyzing the ultrasonographic sign of the 205 breast solid nodular lesions. These lesions were classified according to the total scores of each ultrasonographie sign. ROC curve was used to evaluate the value of UCDSS. Results The area under ROC curve of UCDSS was 0. 977, the sensitivity and specificity was 92.0%, 92.3% respectively. Conclusions The classification based on the UCDSS may increase the diagnostic accuracy of the differentiation between benign and malignant breast solid nodular lesions.

Objective Acute bilirubin encephalopathy in neonates is the most serious complication of neonatal hyperbilirubinemia, is one of the main causes of neonatal death and disability. Clinical early diagnosis, early treatment can improve the prognosis in children. Methods Brainstem auditory evoked potential (BAEF) was detected on two patients (40 patients with ABE, 40 cases of normal controls, all full-term) in the state of sleep in children and analysis the difference between the two groups ,all testing was completed by experienced Department of ENT full-time technician in charge,SPSS15.0 statistical analysis software was took for data analysis (using rank sum test method). Results There was significant difference between the two groups of neonatal latency of wave I, latency of waveⅤ, interpeak time , acute bilirubinⅠ-Ⅴencephalopathy group was significantly longer than that of the control group. Conclusions The BAEF detection is the sensitive index of brainstem damage , can objectively and sensitively reflect the function of the central nervous system , can reflect the functional status of cochlear and brainstem structures , often brainstem was slightly damaged but no clinical symptoms and signs , BAEP has changed significantly , so the conventional BAEP examination performed on patients with hyperbilirubinemia help to find bilirubin brain damage as early as possible,and prevent the occurrence of bilirubin encephalopathy.

OBJECTIVE: To investigate gene-therapy for human hepatocellular carcinoma with adenovirus vectors by double suicide gene CD/TK. METHODS: Double suicide gene CD/TK was liberated from eukaryotic vectors pCEA-CD/TK and subcloned into shuttle vectors, and the transfer plasmid pAdtrack-CMV-CD/TK was formed after linearizing with Pac 1. It was recombinated with pAdeasy-1 in bacteria BJ5183. The identified adenovirus plasmid was digested by Pac1 and was transfected into 293 cells to pack the adenoviruses. After PCR determination, its titre was measured, and the infection rate and efficacy were tested in human hepatocellular carcinoma cells. RESULTS: pAdtrack-CMV-CD/TK and pAd-CD/TK were tested by endonuclease digestion. Ad-CD/TK was produced in 293 cells, and the human hepatocellular carcinoma cells (SMMC7721) infected by Ad-CD/TK were killed after 5-FC was used, and bystander effects were observed. CONCLUSION: Recombinant adenoviruses with CD/TK fusion suicide gene have a high infection rate and efficacy for human hepatocellular carcinoma cells.

Objective To investigate expression of Fos protein in rat brains following restraint stress. Methods The experimental rats were restrained in a small plastic tub for,1,3 and 6 hours,and were sacrificed at 30 min after removing restraint.Immunohistochemical ABC method was used to observe distribution of Fos protein-like immunoreactive(-LI)products in rats brain.Results Fos-LI neurons appeared in (1)Frontal brain:the cingulum,cortex(especially in third and fifth layer),lateral septal nucleus,central amygdaloid nucleus.(2)Diencephalon:the thalamic paraventricular nucleus,lateral geniculate body and medial genicular body,hypothalamic paraventricular nucleus,supraoptic nucleus,periventricular area of third ventricle,arcuate nucleus.(3)Brain stem:the superficial layer of superior colliculus,periaqueductal gray,cortical area of inferior colliculus,lateral parabrachial nucleus,locus coeruleus,A5 area,cochlear nuclei,medullary viceral zone in medulla oblongata.The expression of Fos-LI neurons peaked in rats restrained for 1h,at 3h,then began to decrease,at 6h,significantly decreased. Conclusion Fos-LI neurons appeared in many areas of brain induced with restraint stress.The number of Fos-LI neurons decreased following restraint time.

Objective:To compare the compressive strength of porcelain crown with Co-Cr,CAD/CAMpure titanium and CAD/CAM zirconia respectively.Methods:Metal models of simulating crown core of the ideal premolar were manufactured,digital data of the met-al model were obtained by CAD/CAMsystem,the basement crowns of Co-Cr,CAD/CAMpure titanium and CAD/CAMzirconia were respectively made(n =5),size of each crown was kept the same and the spcimens were set as group A,B and C respectively.Then masking porcelain and body porcelain were fired on basement crowns according to the instructions.The thickness of the porcelain was kept the same.All of the porcelain crowns were located on the Instron testing machine,the compression strength force values were measured.Data were statistically analysed by SPSS 1 3.0 software.The bonding surface of basement material and porcelain was observed by SEMafter compressive failure.Results:The compressive strength(N)of group A,B and C was 2 990 ±1 88,2 305 ±1 57,2 1 50 ± 1 31 ,A vs B or C,P <0.05,B vs C,P >0.05.Conclusion:All the 3 base materials with porcelain crown satisfy the clinical require-ments.The compressive strength of Co-Cr porcelain crown is stronger than that of CAD/CAMpure titanium porcelain crown and CAD/CAMzirconia porcelain crown.

Objective To establish KIF4A 3'UTR esiRNA library and analyze the advantage of method in studying the function of KIF4A in SGC-7901 cells.Methods The GST-fusion protein of Escherichia coli endoribonuclease Ⅲ (GST-RNase Ⅲ) was used to digest the double strand RNA (dsRNA),which was transcribed in vitro from a 502bp template of KIF4A genome (T7-KIF4A 3'UTR).The KIF4A esiRNA library generated from the above method and chemically synthesized KIF4A siRNA were then used to transfect SGC-7901 cells at 10 nmol/L and 20 nmol/L.Real-time quantitative PCR and Western Blot were used to detect the mRNA and protein level of KIF4A,respectively.Results The KIF4A esiRNA library was effectively established from dsRNA digestion using GST-RNase Ⅲ.KIF4A expression was significantly reduced in SGC-7901 cells transfected with KIF4A esiRNA or siRNA.In addition,the inhibitory effect of KIF4A esiRNA was more effective than that of chemically synthesized siRNA.Conclusion KIF4A esiRNA library which was obtained using biological method can more effectively inhibited the expression of KIF4A more effectively in SGC-7901 cells than chemically synthesized siRNA.Therefore,esiRNA library can be used as a new and more effective method in studying the function of KIF4A in SGC-7901 cells.

Objective: To construct a phage display library of human single-chain Fv antibodies against blood group Rh(D) substance. Methods: Combining phage display library techniques, isolated total RNA from B lymphoblastoid cell lines secreting anti-Rh(D) antibodies was used for the synthesis of the first strand of cDNA, V_ H and V_ L genes were amplified by 2nd PCR and linked together by splicing overlap extension (SOE) with the use of a (Gly_ 4Ser)_ 3 linker. The resulted scFv genes were then cloned into pCANTAB5E vectors and displayed on the phage. Phage clones were selected using intact red cells as a source of antigen. After 4 rounds of "binding-elution-enrichment", each clone was assayed for specificity by Dot ELISA. Results: A phage antibody library, with the sink size being 1.2?107, was obtained. The percentage of full-length scFv gene inserted into phage DNA was 0.80. Rescued by helper phage, a phage scFv library with titer of 3?108 pfu/ml was established. Specific phages with scFv were acquired after 4 rounds of panning, one clone exhibiting specific binding to Rh+ cell was identified by Dot ELISA. Conclusion: A strategy for construction phage antibody library by means of phage display technique was practicable, which would be useful in screening engineered antibodies against human Rh (D) blood group substances.