Objective: To determine if the hemagglutinin A (HagA) of Porphyromonas gingivalis could be involved in the adhesion and invasion in human gingival epithelial cells (HGEC). Methods:P. gingivalis 381 hagA mutant was constructed by conjugation method. The whole length of hagA gene was cloned into pYA292 in Salmonella typhimurium x4072 (S. typhimurium-hagA). The strains were used to test their ability of adhesion and invasion into HGEC using a standard antibiotic protection assay. S. typhimurium x4072 strains containing empty vectors were used as negative control. HagA expression in S. typhimurium-hagA was confirmed by Western blot. Results:Although there were no significant differences between P. gingivalis 381 hagA mutant and wild type in adhesion and invasion into HGEC, the adhesion values of S. typhimurium-hagA to HGEC were increased by 3 times compared to their respective controls, while the invasion ability of S. typhimurium-hagA was 4 times greater than that of the negative controls. Conclusion: These results suggest that HagA may participate in P. gingivalis adhesion and invasion into HGEC.

9β,11β-Epoxypregn-4-ene-17α,21-diol-3,20-dione 21-acetate (Ⅰ) is a substrate for the production of 9β,11β-Epoxypregn-1,4-diene-17α,21-diol-3,20-dione (Ⅳ),which is a key precursor for the production of many 9-fluoro-substituted corticosteroid hormones.By comparing whole cells catalysis and cellular lysates conversion,it was found that whole cells of Mycobacterium sp.MS136 could only convert Ⅰ to 9β,11β-Epoxypregn-4-ene-17α,21-diol-3,20-dione (Ⅱ),and Ⅰ can be effectively converted toⅣ by cellular lysates.The reaction order is that Ⅰ is spontaneously hydrolyzed to Ⅱ and Ⅱ undergoes C1,2-dehydrogenation reaction to Ⅳ.In order to improve the productivity of Ⅳ,the key genes kstD,kstD3 and kstDM encoding C1,2-dehydrogenase (KSTD) were overexpressed in Mycobacterium sp.MS136 to enhance the C1,2-dehydrogenation reaction rate,and the results showed that 1 g/L substrate Ⅰ can be converted by recombinant strain MS136-kstDM cellular lysates at pH 7.0,the productivity of Ⅳ reached 78.4％ after 45 h,which is 38.9％ higher than original strain.The reaction rate is enhanced by optimizing the pH,and the results showed that 1 g/L substrate (Ⅰ) can be converted by recombinant strain MS136-kstDM cellular lysates at pH 7.5,the productivity of Ⅳreached 92.8％ after 45 h,which was 63.4％ higher than original strain.

Objective To explore the effects of chronic fluorosis on neurons in the cerebral cortex of rats,and to provide some morphological evidence of damage in the central nervous system induced by chronic fluorosis.Methods Male Wistar rats 40 days after birth were fed with high fluoride contented water(100 mg/L)for inducing chronic fluorosis.Immunocytochemistry and in situ hybridization were used to detect c-fos and Caspase 8 at cerebral cortical neurons respectively.Results c-fos positive cells rate and gray scale in the cerebral cortex of chronic fluorosis were 35.8%and 0.2756±0.0241,respectively,and that of control group were 32.1%and 0.2774±0.0331with statistical difference(χ2=0.305,t=0.826,P＞0.05).Caspase 8 positive cells rates of fluorosis group and control group were 18.7%and 14.1%,respectively,the difference being statistically significant(χ2=0.419,P＞0.05).The gray scale of fluorosis group and control group were 0.3874±0.0329 and 0.3884±0.0323,respectively,the difference being statistically significant(t=0.641,P＞0.05).Conclusion Chronic fluorosis had no significant influence on apoptosis of cerebral cortical neurons.

BACKGROUND: Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Researches suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.OBJECTIVE: To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 (IL-1),IL-6 and tumor necrosis factor alpha (TNF-α) in aqueous humor of rabbits after extracapsular cataract extraction.DESIGN: Randomized controlled animal study.SETTING: Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.MATERIALS: The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes,general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification: SYXK (yue) 2005-006]. Eyes of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawei Biotechnology Company Limited.METHODS: Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mi/kg urethan, and then, 5# needle was punctured from corneal limbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque: horizontally intercepting bladder or waterly separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1st, 3rd, 7th and 14th days after operation, inflammatory reaction of anterior chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meanwhile, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.MAIN OUTCOME MEASURES: Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rabbits in three groups after operation.RESULTS: ① On the 1st, 3rd, 7th and 14th days after operation, numbers of leucocytes were (2.4±0.7)×106/L, (2,2±0.5)×106/L, (2.8±0.8)×106/L and (2.0±0.5)×106/L in aqueous humor; (19.7±7.3)×106/L, (28.1±9.6)×106/L, (14.2±5.6)×106/L and(8.4±3.8)×106/L in traumatic control group; (65.3±14.5)×106/L, (79.8±12.7)×106/L, (21.7±8.2)×106/L and (12.4±4.1)×106/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group (F =22.5, 27.9, 11.6, 8.4;P＜0.05). ② Within 1-14 days after operation, contents of IL-1, IL-6and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group (P＜0.05), and there was a significant difference between traumatic control group and operative group (P＜0.05);however, there was no significant difference among three groups on the operative day (P＞0.05). ③ Contents of cytokines reached peak on the 7th day after operation, decreased gradually, and reached the lowest value on the 14th day.Contents in traumatic control group and operative group were higher than those in normal control group (P＜0.05).CONCLUSION: The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

To evaluate changes in quality- of- life of adults with concomitant exotropia before and after surgery.
●METHODS:A retrospective cohort method was used in this research. Sixty - five patients with concomitant exotropia ( ranged from 18 - 30 years) were enrolled. Quality of life was studied with 2 different questionnaires [the Adult Strabismus - 20 ( AS - 20) and the MOS 36 -item Short - Form health survey ( SF - 36 )], which patients completed preoperatively and at 3mo postoperatively.
●RESULTS: With the AS - 20, 3mo after surgery, the mean psychosocial and visual function scores of AS- 20 improved significantly (P<0. 01). Similarly, with the SF-36, the mean score in 7 of 8 areas improved significantly ( P < 0. 01 ), including physiological function, role limitations due to physiological health, general health, vitality, social function, role limitations due to emotional problems, mental health and the overall mean score. However, with respect to bodily pain, no significant improvement was found after surgery(P>0. 05).
● CONCLUSlON: Surgical treatment of concomitant exotropia in adults gives a highly significant improvement in quality - of - life scores. We should pay more attention to the impact of strabismus on quality of life clinically to improve the outcome of the surgery.

Objective To develop a real-time polymerase chain reaction(PCR)system to determine transcriptional level of MexAB-OprM multidrug efflux pump gene and to investigate the impact of androgra- pholide on MexAB-OprM gene transcription in Pseudomonas aeruginosa.Methods The fragments of mexB gene of mexAB-oprM operon and 30S rRNA gene rpsL were amplified and cloned into two plas- mids respectively.These plasmids were used as external standards for real-time PCR.Real-time PCR was applied to measure the mRNA transcripition of mexB and rpsL gene in Pseudomonas aeruginosa growing in medium with different concentrations of andrographolide.Results The plasmids for standard curve were constructed successfully.The relative mexB mRNA expressions in 50,100,150 and 200?g/mL andrographolide were 0.04?0.03,0.06?0.07,0.09?0.03 and 0.04?0.03 respectively, which were significantly lower than that in the control(0.24?0.04,P0.05).Conclusion Andrographolide can reduce the transcriptional level of MexAB-OprM,which may he one mechanism for its anti-infection effect.

Objective To study effect of eleetromyograghie(EMG) biofeedback training on three kinds of u- rinary ineontifienee.Methods Nineteen patients with urinary incontinence were treated by means of EMG biofeed- back training twice a day for six weeks.The treatment was performed with a device,which can detect the EMG ampli- tude of the pelvic muscle and deliver electric stimulation accordingly.Results After 6 weeks of treatment,the inci- dence of uretbrorrhea was reduced by 41%,and the frequency of micturition was decreased by 38% ,while the fre- quency of urination in one day reduced to 9 to 13.The general subjectively rated improvement rate of patients was 53% ,while the general objectively one was 58%.Conclusion Biofeedback training has significant therapeutic: eftects on patients with urinary incontinence.

Objective To explore a simpler, more economic and reproducible method to reproduce a model of high oxygen induced acute lung injury (HALI) in rats. Methods An animal feeding box equipped with a controllable high oxygen was designed. 100 Sprague-Dawley (SD) rats were divided into normal control group and HALI group by random number table method, with 50 rats in each group. Each group was randomly subdivided into five subgroups according to the duration of exposure to high oxygen, namely 0, 24, 48, 72 and 96-hour subgroups, with 10 rats in each subgroup. The rats in normal control group were kept in cages with ambient air, and the rats in HALI group were kept in an oxygen tank in which the oxygen concentration was higher than 90% volume ratio, with the temperature maintained at 25-27 ℃, humidity of 50%-70%, and CO2 concentration < 0.5% for 23.5 hours every day. The arterial blood of rats was collected for analysis of blood gas at all time points, and the oxygenation index (OI) and respiratory index (RI) were calculated. Then the rats were sacrificed and the right lung was harvested, which was sectioned and stained with hematoxylin and eosin (HE). The changes in histopathology were observed with light microscopy, and pathological score was recorded. The left lung was harvested for the measurement of the wet/dry weight ratio (W/D). Results With the prolongation of high oxygen exposure time, the degree of lung injury in HALI group was gradually increased, and the degree of derangement of alveolar structure appeared in an increasing degree, with destruction of the alveolar wall, widening of alveolar space, and appearance of edema, and inflammatory cell infiltration. A small quantity of red blood cells exudation could be found in some rats. The pathologic changes were most obvious at 48-72 hours after exposure. With the prolongation of high oxygen exposure time (0, 24, 48, 72, 96 hours), the OI (mmHg, 1 mmHg = 0.133 kPa) in HALI group was gradually decreased (446.67±29.93, 306.19±37.23, 269.70±29.00, 253.81±43.40 and 245.58±35.25), RI, pathological score of lung tissue and W/D ratio were gradually increased [RI: 0.25±0.04, 0.31±0.06, 0.38±0.06, 0.46±0.07 and 0.44±0.03; pathological score of lung tissue: 0.00±0.00, 0.90±0.74, 2.90±1.20, 4.70±1.57 and 4.80±1.23; lung W/D ratio: 3.84±0.61, 4.14±0.46, 4.56±0.34, 5.32±0.27 and 5.18±0.25]. Statistically significant differences were found in 72-hour group as compared with that of other groups (all P < 0.05), while no significant difference was found between 96 hours and 72 hours groups (all P > 0.05). There were significant differences in changes between 24, 48, 72, and 96 hours as compared with those of the normal control group: OI (mmHg): 24 h 306.19±37.23 vs. 435.65±25.34 and 96 h 245.58±35.25 vs. 465.42±24.75; RI: 24 h 0.31±0.06 vs. 0.24±0.04 and 96 h 0.44±0.03 vs. 0.24±0.06. The same as true in pathological scores of lung tissue: 24 h 0.90±0.74 vs. 0.00±0.00 and 96 h 4.80±1.23 vs. 0.00±0.00; lung W/D ratio: 24 h 4.14±0.46 vs. 3.79±0.44 and 96 h 5.18±0.25 vs. 4.12±0.91, all P < 0.05. Conclusions A self-designed high oxygen box is simple, easy to operate and reproduction of HALI model can be attained. Sustained exposure to high concentrations of oxygen (≥ 90%) for 24 hours can replicate the HALI model successfully, and the most serious injury appears at 48-72 hours after exposure.

Objective To compare the three kinds of methods for in vitro primary culturing of rheumatoid arthritis synovial fibroblast-like cells (RASFs), in order to get fast and effective culture methods. Methods Synovial tissue from RA synovial arthroscopic resection were treated by collagenase digestion method, modified tissue culture method, double enzyme digestion method respectively. By using an inverted phase contrast microscope, cell morphology and growth characteristics were observed and identified with vimentin staining. Trypan blue was used to count the number of living cells after culturing for 14d. Results The three primary methods could successfully isolate and culture RASFs, and RASFs met the morphological characteristics of vimentin-positive cells>95%, namely, the proportion of RASFs cell confluence was 70% after 16-20days by the collagenase digestion method,whose cell confluence proportion reached 95%after 4 weeks;and the cell confluence proportion was above 70%after 10-14days by modified tissue culture method,and the cell confluence proportion reached 85%after 4 weeks by the double enzyme digestion method. The comparison of the viable cells number cultured same number of synovial tissue by the three methods show the viable cells number cultured by the modified tissue culture method were (1.60±0.08) ×106, those by the collagenase digestion method were (1.41±0.08) ×106, those by the double enzyme digestion method were (1.19 ±0.05) ×106, which were with significant difference among them (P<0.05) .The comparison of incubation time of RASFs primary cells showed it took (267.50±16.58) mins by the collagenase digestion method, (183.75 ±11.08) mins by the double enzyme digestion method, and 149.10 ±13.71mins by the modified tissue culture method, with significant differences (P<0.05) .Conclusion Modified tissue culture for RASFs is an efficient and fast culture method, the number and purity of RASFs can meet the requirements for biology experiments.