Objective:To construct a suppression subtractive library of suppression-related genes from c serotype Streptococcus mutans (S.mutans). Methods:After being isolated from virulent and avirulent strain of S. mutans respectively, the intact and high-pure genomic DNAs were digested with restriction enzyme AluⅠ. The digested DNA of the avirulent strain ligated with an adaptor was used as tester DNA, and that of the virulent strain as driver DNA. Then the suppression subtractive hybridization(SSH) was carried out, the efficiency of ligation and subtraction were detected respectively. The subtracted fragments were inserted into vector pCR2.1 using T/A cloning kit and transformed into E. coli TOP10F' competent cells. The white colonies were selected to construct the suppression subtractive library. Results: Through electrophoresis of AluⅠ-digested DNAs, a smear ranged from 0.1 to 2.0 kb was observed. The ligation efficiency of tester DNA with adaptor was at least higher than 25 percent. The subtraction efficiency confirmed the success in enrichment of differential genes between virulent and avirulent strain of S. mutans. In the subtracted group, the appearance time of the 23S rRNA gene in both tester and driver DNA was later than that in the unsubtracted group by twelve cycles. It suggested that suppression subtractive hybridization happened indeed. Then the subtracted fragments were cloned and the suppression-related gene library between virulent and avirulent strain of S. mutans was constructed. Conclusions:The suppression subtracted library of suppression-related genes has been constructed.

Objective:To explore avirulent strain-specific new genes or new functions of already known genes from Streptococcus mutans of serotype c.Methods:Twenty-six DNA fragments unique to avirulent strain of Streptococcus mutans were sequenced.The sequences of these presumptive avirulence DNA fragments were subjected to homology search through BLASTn and BLASTx in public database,and their putative biological functions were analyzed.Results:The size of the DNA fragments ranged from 113 bp to 776 bp.The average G+C content was 38.27%,similar to that of the gene-codingsequences in Streptococcus mutans strain UA159 whose genome sequences were just completed.Seven clones were picked repeatedly.Of the nineteen DNA fragments,eight potentially represented new DNA fragments were registered in GenBank.The remaining DNA fragments showed high homology to known genes of Streptococcus mutans strain UA159.Conclusions: The gene analysis and identification supply the groundwork for further study of gene functions of the avirulent strain of Streptococcus mutans serotype c.

Objective To detect the expression levels of serum alpha fetal protein (AFP),carbohydrate antigen 199(CA-199) and carcinoembryonic antigen(CEA) in a rat model of liver cancer and elderly liver cancer patients,and explore its clinical significance.Methods A rat model of liver cancer was successfully established by intraperitoneal injection of a low dose of diethylnitrosamine (DEN).The expression levels of serum AFP,CA-199 and CEA in rats were determined and compared during the development of liver cancer.Blood samples were collected from elderly subjects with normal liver and elderly liver cancer patients(n=80,each).The expressions of serum AFP,CA-199 and CEA were determined and compared between healthy elderly patients and elderly liver cancer patients.Results Serum levels of AFP,CA-199 and CEA were higher in liver cancer rats than in normal rats [(4.21±1.32) μg/Lvs.(1.05±0.33) μg/L,(3.78±1.04) kU/L vs.(1.00±0.28) kU/L,(3.54±0.92) μg/Lvs.(1.10±0.37) μg/L,t=9.493,8.609,7.675,respectively,all P=0.000].Serum AFP,CA-199 and CEA levels were higher in elderly patients with liver cancer than in elderly subjects with normal liver [(66.89±9.33) μg/L vs.(2.56±1.09) μg/L,(116.89±43.33) kU/L vs.(5.56±1.26) kU/L,(5.83 ± 1.56) μg/L vs.(1.17 ± 0.51) μg/L(t=14.379,17.470,10.677,respectively,all P=0.000).The positive rate of joint detection of AFP,CA-199 and CEA was higher in elderly patients with liver cancer than in elderly subjects with normal liver [77.50％ (62/80) vs.1.25％(1/80),x2 =17.260,P=0.000].Conclusions The joint detection of AFP,CA-199 and CEA can help increase the diagnostic rate of liver cancer in elderly patients.

[Objective]To establish a method to determine the contents of glucase and fructose in extracts of leaves of Lonicera hypoglauca Miq. using HPLC-ELSD, and apply this method to study the content change of glucose and fructose in different solvents extracts in leaves of Lonicera hypoglauca Miq. [Methods] The chromatographic separation was achieved on Lichrospher NH2 column using a mobile phase composed of a mixture of acetonitrile and water(90:10) at a flow rate of 0.8mL·min-1 and the column temperature was 25℃.The ELSD drift tube temperature was set at 70℃ and the carrying gas was N2 and the flow rate of the gas was 1.6L·min-1. [Results] Glucose and fructose from extracts of the leaves of Lonicera hypoglauca Miq. could be wel separated, and the linearity ranges of glucose and fructose were shown at the concentration of 0.5840~1.168mg·mL-1 and 0.8256~1.8576mg·mL-1, the average recovery rate of glucose and fructose were 96.88%(RSD=1.11%) and 97.55%(RSD=1.15%), the stability of glucose and fructose was 2.12%and 0.38%, and the repeatability was 1.31% and 0.24%. Among the water extract,50% alcohol extract and 95% alcohol extract,the content of the glucose and fructose in the water extract was highest,which was 2.19mg·mL and 4.10mg·mL-1, the 50% alcohol extract was second, which was 1.13mg·mL-1 and 2.12mg·mL-1, and the 95% alcohol extract was lowest, which was 0.45mg·mL-1 and 0.65mg·mL-1.[Conclusion] The method is simple, accurate and has good repeatability, and can be used to determine the contents of glucose and fructose in the extract of leaves of Lonicera hypoglauca Miq. In this batch of leaves of Lonicera hypoglauca Miq., the contents of fructose and glucose were of water extract>50%ethanol extract>95%ethanol extract.

Objective To investigate the relationship among the proliferation of CD4+T cells, the intracellular levels of interleukin-10 (IL-10) and transforming growth factor-?1(TGF-?1), and allergic bronchial asthma. Methods Dermatophagoides farinae antigen were prepared as allergen. Twenty-five patients with asthma and 15 healthy individuals were enrolled and divided into blank control group, allergen group and self control group, respectively, after venous blood sample collection. The proliferation of CD4+T cells and the distributions of CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 were measuredby flow cytometry (FCM). Results The distributions of CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 could hardly be detected in the peripheral blood samples of the blank controls of the patients with asthma and healthy ones. In the allergen group of the healthy individuals, the peripheral blood CD4+T cells were significantly proliferated, and the proportions of CD4+T cells andCD4+/IL-10+ cells were much higher than the self control group, while there was no significantly increase in the proportions of CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 subgroups. In the allergen group of those with asthma, the proportions of peripheral blood CD4+, CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 cells were not found significantly increased compared with those self controls. After being activated by allergen, the proportion of peripheral blood CD4+/IL-10+ cells was significantly lower in the patients with asthma than the healthy individuals(P

Objective To compare the diagnostic value of CT and NE for lesions of nasal cavity and paranasal sinus area.Methods 80 cases of nasal and paranasal sinus diseases which were examined by both CT and NE at same period and proved by pathology.were reviewed.Results Compare with results of pathology,the accucy rate of both CT and NE was identical in diagnosing sinusitis and tumours of paranasal sinus,NE was higher than CT in nasal and paranasal sinus polyp and nasal cavity tumour,turbinate hyperplasia CT was higher than NE in paranasal sinus cyst and anatomic variations.Conclusion Pathologic biopsy can be taken under NE guide to understand the shape,texture and colour of the illness focus CT can show the size,shape and anatomy relation between lesion and its neighbor tissue.The diagnosis may more perfectly through NE combined with CT.

Objective To investigate radiation dose diffent type of anesthesia affected radiation dose when we do interventional therapy for child patent ductus arteriosus (PDA).Methods From 201 1 to 2014,we collected 50 children(3-6 years old)with PDA which received interventional therapy were included in the study.They were divided into two groups:group A (25 cases,male/fe-male=7/18,mean weight=1 5.32 kg±2.41 5 kg)underwent interventional therapy of PDA under general anesthesia,and group B (25 cases,male/female=13/12,mean weight=1 6.40 kg±2.056 kg)using local anesthesia.The surgery were operated by the same doctor,we used DSA children cardiovascular film AE mode (ped CARD)to monitor the surgery,image frames 1 5-30 f/s,used the non-ionic contrast agent (Iodixanol 320 mg I/mL)and recorded the child cumulative incidence of skin surface dose (AK),dose area product(DAP),and time of fluoroscopy,and do statistical analysis.Results All 50 cases were performed the surgery successfully. There were no significant difference of age,weight and gender between groups (age:t=1.924,P =0.06;weight:t =1.703,P =0.095;gender:χ2 =3.00,P =0.083).The cumulative incidence of skin surface dose (AK),dose area product (DAP)and time of fluoroscopy were (0.061±0.025)Gy,(5.08±2.19)Gy·cm2 and (3.15±1.16)min in A group,and (0.094±0.046)Gy,(8.41±3.587)Gy·cm2 , (6.86±3.27)min in B group.The sequence and image number of two groups were same.There were significantly differences of cumulative incidence of skin surface dose (AK),dose area product (DAP)and time of fluoroscopy between two groups (AK:t =3.152,P =0.003;DAP:t =3.957,P =0.000;time of fluoroscopy:t =5.346,P =0.000).The radiation doses of A group significantly lower than B group,compared with B group,the radiation dose were 40 percent [(1-5.08/8.41)%]lower in A group,the 1.1 7 times discrepancy of time of fluoroscopy between two group [(3.1 5-6.86)/3.1 5].Conclusion Radiation dose is associated with type of anesthesia. Compared with local anesthesia,radiation dose reduced 40% using general anesthesia during interventional therapy for PDA.

Objective To observe the formation of biofilms on endotracheal tubes ,and to determine the effect of azithromycin on Haemophilus influenzae in biofilms .Methods One hundred and twenty Wistar rats were intubated with a silicone tube ,80 of them(40 rats were model group and 40 were medicine group) were infected by bacteria to establish the animal models of endotra-cheal tube infections the others were ues as control gruop .Formation of biofilms was detected by scanning electron microscope .Via-ble cells counting was carried out to evaluate the effect of azithromycin orally on biofilms on endotracheal tubes .Results Uneven biofilms were formed on the surface of endotracheal tubes ,and slim bodied of bacteria enclosed by a lot of extracellular matrix .Via-ble cells counting was reduced in medicine group compared with model group .Conclusion Haemophilus influenzae can form bio-films on endotracheal tubes .Azithromycin can exert inhibitory effect on haemophilus influenzae biofilms formed on endotracheal tubes .

BACKGROUND: Many scholars have paid attentions on neural stem cells (NSCs), which are regarded as the cell source to repair the injured nervous tissues. And it is becoming a key point that NSCs will be obtained in an effective way.OBJECTIVE: To observe the characteristics of isolation, proliferation and differentiation of in vitro NSCs from subventricular zone (SVZ) and hippocampus of neonatal rats.DESIGN: Single-sample trial.SETTING: Department of Neurosurgery in the Third Affiliated Hospital of Sun Yat-sen University.MATERIALS: Ten SD rats aged 3 days, of either gender, were offered from the Experimental Animal Center of Sun Yat-sen University. Nestin antibody (rabbit-anti-rat), neurofilament (NF-200) antibody (rabbit-anti-rat), and glial fibrillary acidic protein (GFAP) antibody (mice-anti-rat) were all purchased from Sigma Company.METHODS: The experiment was carried out in the Laboratory of Histology and Embryology, Basic Medical College of Sun Yat-sen University from September to December in 2006. According to the ethical requirement, the serum-free DMEM/F12 medium containing basic fibroblast growth factor and epidermal growth factor was utilized to isolate and incubate NSCs from SVZ and hippocampus of neonatal rats. Then the proliferation and differentiation of NSCs in vitro were observed.MAIN OUTCOME MEASURES: Fluorescence immunocytochemistry was applied to detect the antigen of nestin expressed from NSCs, NF expressed from neuron cells and GFAP expressed from astrocytes. The differentiation of NSCs was identified.RESULTS: The cells isolated from SVZ and hippocampus of neonatal rats possessed the ability of proliferation and self-renewal. Through inverted microscope, nestin positive cells could be found in the cell clone sphere at passage 20 of cells. Cells induced to differentiate could express antigen of NF and GFAP by the attached cell clone sphere.CONCLUSION: The NSCs successfully isolated from SVZ and hippocampi of neonatal rats possess the ability of self-renewal, proliferation, and differentiation into neuron-like cells and astrocytes.

Objective To investigate the value of soluble epithelial cadherin(sE-cad)in the differential diagnosis of pleural effusion. Methods Patients were divided into malignant pleural effusion group,infective pleural effusion group and transudation group.sE-cad in pleural fluids obtained during the first thoracocentesis was measured by enzyme-linked immunosorbent assay(ELISA).The concentration of sE-cad in all kinds of pleural effusions was compared.The cut-off value of sE-cad for the differential diagnosis of benign and malignant pleural effusion was determined by ROC curve.The diagnostic value of sE-cad was also compared with common tumor markers such as CEA,CA199,CA125 and NSE.Results The concentration of sE-cad was significant higher in the malignant pleural effusion than in the benign pleural effusion[(38.38?4.15)ng/mL vs(14.17?0.80)ng/mL,P