OBJECTIVE To explore the role of Tn5281 in high-level gentamicin resistance(HLGR) of Enterococcus faecalis. METHODS Fifty four strains of HLGR E.faecalis were tested for Tn5281 by PCR method.The PCR products were confirmed by dot blot hybridization and sequencing.Filter mating method was used to investigate the conjugative plasmid transfer.Antimicrobial susceptibility test was performed by agar dilution method of CLSI/NCCLS. RESULTS Of 54 HLGR E.faecalis strains,27(50.0%) carried different types of Tn5281,with 6 strains carrying complete type,17 strains carrying truncated type Ⅰ(lacked IS256 in the right flanking extremity) and 4 strains carrying truncated type Ⅱ(lacked IS256 in the left flanking extremity).All types of transposons were loaded on plasmids. Among four HLGR E.faecalis in which Tn5281 was successfully transferred to recipient strains by filter mating method,one carried complete type and three carried truncated type Ⅰ.Tn5281 carrying strains had higher resistance rates to penicillin and ampicillin and lower resistance rates to tetracycline and chloramphenicol compared to the strains not carrying Tn5281.(CONCLUSIONS Tn5281) is commonly carried by HLGR E.faecalis.It may integrate into conjugative plasmids and play an important role in the dissemination of HLGR among E.faecalis.

Objective To determine the relationship between endothelin-l(ET-1)with blood flow of gastric mucosa and gastric mucosal injury (GMI) in obstructive jaundice rats induced by coldly restrainting stress. Methods 161 rats were randomly assigned into the experimental (obstructive jaundice) group (n=70), the control (sham operative) group (n=70) and the anti-ET-1 preconditioning group (n=21). The obstructive jaundice model was set up in the experimental group and the anti-ET-1 preconditioning group. Both the experimental group and the control group were re-assigned into 5 subgroups which underwent non-coldly restraint and coldly restraint for 10, 30, 60 and 120 min respectively. The anti-ET-1 preconditioning group underwent anti-ET-1 serum precondition 30,60 or 120min before the coldly restraint. The ET-1 was detected by radioimmunologic method and the gastric mucosal blood flow (GMBF) by laser Doppler flowmeter. Results The concentrations of ET-1 in gastric mucosa and ulcer index significantly increased (P

Although there are numerous case resource,there still exist a relatively long distance in clinical research in China in comparison with developing countries because of inadequate recognition and funding,which has greatly affected the development of standardization of clinical medical in China for the past few decades.With the deepening of recognition and increasing of special support to clinical medical research in China,the development based on clinic research will face new opportunities.As a result,it is necessary for ophthalmologists to explore clinical issues,leverage from the mega trend of clinical research and deliver high-quality results.To achieve this goal,ophthalmologists will have to constantly update their knowledge base,standardize clinical practice and improve diagnosis methods.So far,challenges persist in clinical research in ophthalmology as follows:It is difficult to get funding for clinical research project;It is difficult to get ethnics approval for clinical research project;The research design for forward-looking projects is yet to be standardized;Cross-disciplinary collaboration needs to be enhanced.This review was expected to inspire more attention from clinical research specialists and consequently more highquality clinical studies.

Objective　To study the ultrasound image feature of acute hepatic lesions in different times.Methods　The feature of ultrasonography in 126 cases with acute hepatic lesions were analyzed and divided into two groups according to different fime after injury: group A, 25～60 minutes after injury; group B, 60～120 minutes after injury.Results　There was significant difference in the structure of sonographic classification between two groups. The high echos was obviously higher in group A than that in group B, while the mixed echos was obviously higher in group B than that in group A (P<0.001). Compared each types in group A and in group B,the high achos in group A and the mixed achos in group B were higher than other types in same group (P<0.001).Conclusions　The ultrasonographic feature of acute hepatic lesions would change with time. The high echos occurs in eaily time after injury ,and then changes into the mixed echos.

To investigate the preventive and treated effects of protein A immunoadsorption on hyperacute rejection of highly sensitized renal transplant recipient. A total of 12 highly sensitized renal transplant recipients were selected from Department of Organ Transplantation, Zhengzhou People's Hospital between January 2006 and January 2008. Immunoadsorption treatment was performed on 12 renal transplant recipients whose panel reactive antigen was more than 50% once a week for 4-6 weeks before operation. At one day before operation, the treatment was performed once. The treatment was performed at once if the acute rejection occurred after operation. The acute rejection and adverse effect were observed. Hyperacute rejection was not occurred in any recipient. Sudden oliguria and increase of serum creatinine were occurred in two cases at several days after operation. The pathological examination showed there were inflammatory cells infiltration and edema in renal interstitial, lymphocyte aggregation around renal tubular, monocytes infiltration in capillary, endarteritis, and necrosis of small vessels in the two cases. They were diagnosed as acute rejection. Their panel reactive antibody level increased again. Average level of Class Ⅰ was 3.8% and class Ⅱ was 19.2%. Acute rejection were reversed and renal function recovered to normal by the therapy of protein A immunoadsorption and pulse therapy of high dose methylprednisolone. Average level of Class Ⅰ was 8.3% and class Ⅱ was 5.2%. One year follow-up were obtained from all recipients. Their transplanted renal function were good, which suggested protein A immunoadsorptin could eliminate antibody in recipients quickly, decrease panel reactive antibody level, and prevent acute rejection.

BACKGROUND: Soluble dispersive mixture of domestic chicken collagen type Ⅱ (SmCC Ⅱ ) has been proven to be safe and effective for rheumatoid arthritis (RA) treatment while there are some similar articular cartilage degradation and autoimmune pathogenesis between osteoarthritis (OA) and RA, so it is worth studying whether SmCC Ⅱ is effective for the precaution or treatment of OA.OBJECTIVE: To observe the prophylactic and therapeutic effects of SmCC Ⅱ on rat OA and analyze the concomitant matrix metalloproteinase (MMPs) and tissue protease changes in osteoarthritic rats.DESIGN: Randomized and controlled observation.SETTING: Department of Clinical Pharmacy, Sixth People's Hospital, Shanghai Jiao Tong University.MATERIALS: Totally 258 rats of clean grade and either gender, aged 6 weeks were involved in this experiment. SmCC Ⅱ (white powder, Batch No.00031004) was provided by Professor Ren Geng-Fu from Shanghai Engineering Institute of Herbal Biomedicine. The 20 mg/L and 80 mg/L solution of SmCC Ⅱ effective component were prepared before use.While 0.25% excipient (mannitol, product of Jiangsu Tianyuan Medical Co., Ltd) solution was prepared by 200 g/L mannitol dissolved in sterile saline solution.METHODS: The study was carried out in the Whole Animal Laboratory and Experimental Center, Sixth People's Hospital,Shanghai Jiao Tong University between March 2003 and February 2006. ①Prophylactic study: Totally 132 rats were randomized into 5 groups: OA group (n =36): treated with sterile saline solution orally by a 16G syringe, 1mL/d; Low- and high-dose SmCC Ⅱ groups (n =24, respectively): treated with 20 mg/L and 80 mg/L SmCC Ⅱ solution orally, 1 mL/d;Excipient group (n =24): treated with 2.5g/L mannitol, 1 mL/d. OA models were surgically induced in these 4 groups by Hilth's method; Normal group (n =24): Rats without operation were treated with sterile saline solution orally, 1 mL/d. All the rats were administrated on the day of modeling. ②Therapeutic study: Totally 126 rats were randomized into 5 groups, grouping, administration and rat number in different groups were the same as those in prophylactic study,respectively, except for the n =30 in OA group. In addition, all the rats were adminstrated for 8 weeks since 6 weeks after operation. The knee joints of right hind limb of all the rats were taken off after 1-week treatment. And sample was cut open along coronal incision for use. ③Morphological study of articular cartilage was conducted by HE staining and Mankin score was calculated to evaluate the severity of arthritis, immunohistochemical studies of matrix metalloproteinase (MMP)-13, MMP-9 and cathepsin K (Cath K) were carried out by ABC method in situ and the percentage of positive-stained chondrocytes were calculated while the mRNA level for MMP-13, MMP-9, Cath K and the tissue inhibitor of metalloproteinase 1 (TIMP1) were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method.MAIN OUTCOME MEASURES: ①The morphological changes of articular cartilage in different groups in prophylactic or therapeutic study.②The level of MMP-13, MMP-9, Cath K and their corresponding mRNA levels.RESULTS: All the 258 rats were involved in the final analysis. ①Prophylactic study: Apparent degeneration appeared in the rats of OA group and the mankin score in OA group was higher than that in high- or low- SmCC Ⅱ groups [(6.44±0.81), (2.75±0.85), (2.70±0.81) points respectively, P ＜ 0.05], the mRNA and positive-stained chondrocyte percentage of MMP-13, MMP-9 and Cath K in OA group was higher than that in high- or low-dose SmCC Ⅱ group, respectively (P ＜0.05-0.01) while the TIMP1mRNA level in OA group was not significantly higher than that in high or low SmCC Ⅱ group (P＞ 0.05). There were no significant changes on the level of MMP-13, MMP-9 and Cath K between excipient and OA group (P ＞ 0.05). ②Therapeutic study: The Mankin's score of OA group was higher than high or low SmCC Ⅱ group [(6.96±1.02), (3.08±0.45), (4.00±0.94) respectively, P ＜ 0.05-0.01] while the mRNA and positive-stained chondrocyte percentage of MMP-13, MMP-9 and Cath K in OA group was higher than that in high- or low-dose SmCC Ⅱ group,respectively (P ＜ 0.05-0.01) while the TIMP1mRNA level in OA group was not significantly higher than that in high- or low-dose SmCC Ⅱ group (P ＞ 0.05).CONCLUSION: SmCC Ⅱ can delay the degradation of articular cartilage of OA rats and maybe effective for OA prevention or treatment. The mechanism maybe related to SmCC Ⅱ reducing the synthesis of MMP-13, MMP-9 and Cath K at transcriptional level.

ObjectiveTo investigate the effect of thalidomide on the proliferation and apoptosis in human pancreatic cancer cell SW1990 in vitro.MethodsSW1990 cell line was treated with thalidomide at different concentrations (3.125,6.25,12.5,25,50,100,200 and 400 μg/ml) for 24,48,72 h,and then cell proliferation were evaluated by MTT.Cell cycle was determined by flow cytometry,apoptosis was determined by annexin V/PI fluos staining,Bcl-2,Bax protein expression and Bcl-2/Bax ratios were measured by Western Blot in vitro.ResultsThalidomide inhibited the proliferation of SW1990 cells in a time and dosedependant manner.The proportion of G0/G1 phase of SW1990 cells with 200 μg/ml thalidomide treatment increased from (41.15 ± 2.23 ) ％ to (58.83 ± 2.33 ) ％,apoptosis rate increased from 2.6％ to 28.0％,the expression of Bax protein up-regulated from 0.17 ± 0.03 to 0.33 ± 0.04,the expression of Bcl-2 protein downregulated from 0.35 ± 0.02 to 0.17± 0.01,the ration of Bcl-2/Bax decreased from 2.17 ± 0.44 to 0.52 ±0.07.ConclusionsThalidomide can inhibit the proliferation of pancreatic cancer SW1990 cells by upregulating Bax,down-regulating Bcl-2,inducing cell apoptosis and cell Go/G1 phase arrest.

Objective To observe the influence of interleukin-1β (IL-1β) on the expression of phosphorylated signal transducers and activators of transcription 3 (pSTAT 3) in rat retinal MOiler cells. Methods For in vitro study cultured Mailer cells were treated with IL-1β of different concentrations (0, 0.1, 1, 5 and 10 ng/ml) for 24 hours. For in vivo study, 32 Sprague-Dawley(SD)rats were divided into 4 groups randomly (control group, 100,500 and 1000 ng/ml group) with 8 rats in each group. After 24 hours of injection with phosphate buffered solution (PBS), or 100, 500, 1000 ng/ml IL-1β into the vitreous treated retinas were evaluated by indirect immunofluorescence and western blotting. Results After 24 hours of incubation without IL-1β, pSTAT3 has little expression in cultured Muller cells, but was up-regulated by 1 ng/ml or higher IL-1β in a dosage-dependent manner (F=46.64, 43.78; P<0.01). pSTAT3 was not expressed in adult rat retina, but was up-regulated by vitreous injection of 100 ng/ml or higher IL-1β in a dosage-dependent manner (F=73.53, 43.70; P<0.01). pSTAT3 expressed mainly in inner nuclear layer and ganglion cell layer. Double-labeling showed that there was no co-staining of pSTAT3 and glial fibrillary treated with IL-1β. Conclusions Expression of pSTAT3 in MUller cells could be activated by IL-1β which may represent one pathway link to reactive gliosis.

Objective To investigate the risk factors associated with early mortality of the rapid two-stage arerial switch operation, which has a significantly higher overall mortality than that of ASO procedure for D-TGA with a intact ventricular septun. Methods The data we reviewed involving patients who underwent rapid two-stage switch operations from September,2002 to September, 2007 in our center, 13 patients were male and 8 were female, their age at operation ranged from 29 to 250 days afer birth, and the body weight was 3.5 to 7 kg. Chi-squared test and multivariant logistic regression methods were used for the analysis of demographic data, pre-diagnosis information, operation data, interval data combined with the operation time,left ventricle training condition, and the early post-operative outcomes. Results The operative mortality was high at initial stage, and then decreased gradually. The logistic multivariant regression analysis indicated that the mortality of left ventricular training operation was associated with the diameters of BT shunt ( P =0.003 ); the mortality of two-stage switch was associated with feminie ( P = 0.006 ) and pre-operative p. LV/RV ( P ＜ 0.001 ). Conclusion Patients with transposition of the great arteries and intact ventricular septum who missed the optimal time for switch operation should receive rapid two-stage switch operations, which provide an opportunity for the correction of the deformity. The key factor associated with the success for operation was good in heart function after left ventricular training.

0.05). ②Therapeutic study: The Mankin's score of OA group was higher than high or low SmCCⅡ group (6.96?1.02), (3.08?0.45), (4.00?0.94) respectively, P 0.05). CONCLUSION: SmCCⅡ can delay the degradation of articular cartilage of OA rats and maybe effective for OA prevention or treatment. The mechanism maybe related to SmCCⅡ reducing the synthesis of MMP-13, MMP-9 and Cath K at transcriptional level.