Objective To investigate the influence of plasmid spread and ampD mutation to Enterobacter cloacae that leads to the AmpC β-lactamase change from inducible type to constitutive type. Methods The Enterobacter cloacae were isolated from the patients with nosocomial infection. The inducible type isolations and their constitutive type changers were put into the same group. The plasmid ampC gene and chromatin ampD gene in pairs in each group were amplified, sequenced and compared. Results Of 195 patients infected by Enterobacter cloacae of inducible type, 25 (12.82%) were changed to the ones of constitutive high type. In these 25 changed groups, 10 were caused by plasmid spread, 10 by ampD mutation, 1 by both, and 4 by neither. Twelve changed constitutive type strains had ampD significant mutations, in which 7 were frame-shift mutations and 5 were spot mutations. Conclusions The change ratio of Enterobacter cloacae from inducible type to constitutive type is rather high. Both plasmid spread and ampD mutation are possibly the mechanism of such change. Plasmid mediated AmpC β-lactamase spreads among different species and interregionally. The mutation rate of chromatin ampD gene is also higher than the natural mutation rate. These two mechanisms should be considered in clinical treatment.

Objective To reveal the influence factor of ampD specific mutation on the transformation of AmpC β-lactamase from non-high-productive type to derepressed the high-productive type in Enterobacter cloacae. Methods The selected cbromatin-mediated derepressed high-productive strains of Enterobacter cloacae and genomic DNA were extracted, and genomic ampD was amplified by using PCR, and then sequenced and confirmed by specific mutation. Use site-directed mutagenesis to mutate the wild-type Enterobacter cloacae for the above site. The changes of AmpCβ-lactamase types were detected by the improved Cefotaxime three-dimensional test. Results Of the 121 strains of Enterobacter cloacae,15 strains were ehromatin-mediated derepressed high-productive type,8 strains had significant mutations and 7 strains of site-directed mutagenesis were established. Ecl MA(274 inser-ting A)、Ecl MC(327 missing C)and Ecl MF(27 inserting G)were changed from non-high-productive type to derepressed-high-pro-ductive type,but Ecl MB(371 inserting T), Ecl MD(515 missing C), Ecl ME(324 C→A), Ecl MG(238C→A 302T→A)did not change in AmpC β-lactamase types. Conclusion The significant mutations that could lead to the transformation of AmpC β-lacta-mase were the Frame-shift mutations that could create the change of large region of amino acid. However, other mechanisms for the derepressed high-productive AmpC β-lactamase were not excluded in the study.

SUMMARY A 61-year-old male patient, with the history of premier myocardial infarction and CABG surgery, experienced recurrent unstable angina. Angiography showed triple vessel disease and vein grafts obstruction. The patient underwent a re-do OPCAB + TMLR + bone marrow mononuclear cell (BM-MNCs) transplantation on Nov. 8, 2004. BM-MNCs were isolated with standard gravity gradient method and the final implanted cell number was 12.06?108. Peri-operative data were similar to those of single OPCAB or re-CABG. The patient recovered promptly without recurrence of angina or infarction. Six-minute walking distance increased significantly (366 m to 493 m). Several imaging examinations reveal improved left cardiac function (LVEF improved from 23.75% to 52% in MRI) and diminished MI area. The results reveal that bone marrow cell transplantation, combined with TMLR and OPCAB is safe and might be effective in improving heart function for patients with IHD.

Objective To observe the clinical effect of emergency operative treatment on thoracolumbar compression or burst fractures using pedicle screw instrument through posterior approach and indirect reduction.Methods53 acute thoracolumbar compression or burst fractures cases injured within 4 days were accepted the emergency operation of indirect reduction, decompression and fixation using pedicle screw instrument through posterior approach. The mean percentage of traumatic vertebral body height, sagittal diameter of spinal canal, degrees of kyphosis and Frankel scoring of nerve function were evaluated before and after operation.ResultsAll cases received two weeks following-up, 47 cases had more than one-year following-up. The mean percentage of traumatic vertebral height restored from 42.28% (pre-operation) to 93.46% (post-operation); the mean degrees of kyphosis recovered from 26.44° (pre-operation) to 17.73° (post-operation); the mean percentage of sagittal diameter of spinal canal changed from 63.14% (pre-operation) to 78.55% (post-operation). All but cases with completely spinal cord injury got more than one grade nerve functional restoration. Neither later spinal cord injury nor back pain existed among the patients. Three cases emerged partial internal fixation device failure, but neither symptoms nor abnormality correction missed.ConclusionEmergency operation using pedicle screw instrument through posterior approach is a feasible and confirmatory choice in treatment of the acute thoracolumbar compression or burst fractures if the cases are selected properly.

?AlM:To understand the clinical effect of trabeculectomy with mitomycin and interferon therapy on neovascular glaucoma.? METHODS: Neovascular glaucoma patients in our hospital from January 2011 to January 2013 were sampled, from them, 57 cases ( 57 eyes ) were randomly divided into two groups, control group received routine trabeculectomy for treatment, the experimental group accepted combination therapy of trabeculectomy +mitomycin + interferon. The clinical efficacy of two groups were observed.?RESULTS:The total effective rate (96%) in experimental group was significantly better than that in the control group ( 76%) , and the visual acuity, filtering bleb and postoperative 1wk, 1a of intraocular pressure changes were better than those of the control group, there was a statistically significant difference (P<0. 05). The rate of complications in experimental group ( 10%) was slightly higher than that of the control group (7%), but there was no statistically significant difference (P>0. 05).? CONCLUSlON: Trabeculectomy combined mitomycin and interferon treatment of neovascular glaucoma has exact clinical effect, and is worthy of clinical popularization and application.

Purpose To investigate the expression of autophagy-related protein LC3B in cervical squamous carcinoma and its relation-ship with Ki-67 expression. Methods To detect the expression of LC3B in 16 cases of normal cervical tissues and 126 cases of squa-mous cell carcinoma by immunohistochemical staining. In addition, Ki-67 protein was also detected in 126 cases of squamous cell car-cinoma in the same assay. The relationship between LC3B expression and Ki-67 in cervical squamous cell cancer was statistical analy-sis. Results Expression level of LC3B were significantly lower in cervical squamous carcinoma than normal squamous epithelial cells (P<0. 05), and the expression of LC3B was negatively correlated with Ki-67(rs = -0. 248, P<0. 05). Conclusion It appears that decreased levels of autophagy which indicated by low expression of LC3B may promote cancer cell proliferation in the early stages of cervical squamous cell carcinoma, which provides a clinical referential evidence for further explore the mechanism of autophagy in cer-vical cancer development.

Objective:To establish an HPLC method for determining four constituents ( schaftoside, isoschaftoside, deoxyelephan-topin and 4,5-dicaffeoylquinic acid) in Shennongcha granules. Methods:An HPLC method was performed on a Hypersil C18 column (200 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile as the phase A and 0. 025 mol·L-1 phosphoric acid solution as the phase B with gradient elution. The flow rate was 1. 3 ml·min-1 . The detection wavelength was set at 270 nm for schaftoside and isos-chaftoside, 208 nm for deoxyelephantopin and 327 nm for 4, 5-dicaffeoylquinic acid. The column temperature was room temperature. Results:The calibration curve was linear over the concentration range of 5. 850-117. 000 μg·ml-1 for schaftoside, 4. 650-93. 000 μg ·ml-1 for isoschaftoside, 4. 160-83. 200 μg · ml-1 for deoxyelephantopin and 5. 470-109. 400 μg · ml-1 for 4, 5-dicaffeoylquinic acid. The correlation coefficient of all curves was more than 0.999. The average recoverywas 97.70% (RSD=1.40%), 96.87%(RSD=1.13%), 97.53%(RSD =1.69%) and 99.29%(RSD =1.01%) (n =6) , respectively. Conclusion: The developed HPLC method is simple,accurate,and can be used in the content determination of Shennongcha granules.

Objective To study the clinical evaluation of gas exchange impairment in neonatal respiratory failure.Methods Blood gas, PaO2/PAO2, Qs/QT, PaO2/FiO2, A-aDO2 and RI of 53 newborn infants with respiratory failure in NICU of PUMCH from Jan. 1993 to Dec. 1997 were measured. Results These infants were divided into two groups according to PaO2/PAO2: moderate and severe respiratory failure. Qs/QT(11±3)%, PaO2/FiO2(183±113), A-aDO2(22.9±6.8)kPa, RI(2.5 ±0.8) in 21 neonates with moderate respiratory failure; Qs/QT(24±6)%, PaO2/FiO2 (82±30), A-aDO2 (49.3 ± 17.8)kPa,RI(7.6 ±3.4) in 32 neonates with severe respiratory failure(P＜0.001). These results in respiratory failure caused by different pathogenesis were different. In meconium aspiration and pneumothorax group there were the highestQs/QT(32±3)% ,A-aDO2 (69.8 ± 12.2)kPa,RI(9.2 ±2.9)and the lowest PaO2/FiO2 (77±39). Mortality of infants with high pulmonary shunt was high. Qs/QT(17±8)% in 38 survives and (24±6)% in 10 died neonates( P＜0.05). Conclusion The clinical evaluation using these indexes for newborn infants with respiratory failure is beneficial in recognizing pathogenesis,guiding therapy and evaluating prognosis.

Objective: To explore the characteristics of IR spectra of the crude drug Herba Sedi collected in different habitats and seasons.Methods: The infrared spectra of all the samples of the whole plants were obtained by FT-IR spectroscopy,the sample powder tablets were used,and identifi cation features were determined through comparative study.Results: The harvest seasons of the samples are remarkly related to the positions(cm-1),intensity or ratios of some major absorbing peaks of the infrared spectra from the 15 samples of several different months and 4 habitats in Hubei Province,China.As for peak positions,the peak values(1625?7)cm-1 of all the samples in spring are less than 1623cm-1,but in autumn(April to October) are generally more than 1628 cm-1.And peak values(1057?16)cm-1 of the samples of spring are more than 1050 cm-1,but less than 1050 cm-1 for the samples of summer and autumn.Moreover,the habitats of samples are also related to IR spectra to some certain extent.Conclusion: The harvest seasons of Herba Sedi from Hubei Province can be preliminarily ascertained according to the IR spectra features.IR technique should be paid attention to in research and identifi cation of the harvest periods of crude plant drugs.

Objective To introduce a method to aid dental treatment for autistic children; midazolam oral sedation. Methods 10 autistic children who can not be managed under normal dental procedures were administrated oral midazolam (0.5mg/kg) before treatment. Once they were adequately sedated, dental treatment was carried out in outpatient. During the whole course, their finger oxygen saturation and heart rate were inspected. Painless methods were also applied. The duration of each treatment was controlled within 30min. When the procedure ended, the autistic children were inspected until recovery from sedation and then discharged with their parents. Results 10 children aged 4～10y totally underwent 17 visits to complete all dental treatment. The average ages were (6.00?1.89)y, body weight was (28.10?9.36) kg and Houpt scales were (4.71?0.77). They were all successfully sedated and managed and no severe complications were found. Conclusion The oral midazolam sedation showed satisfactory effects on this group of autistic children while studies of large samples are needed to draw a definite conclusion.