To improve bond selectivity of recombinant β-glucuronidase in Escherichia coli (PGUS-E), based on the PGUS-E structure guidance, three key points R329, T369 and N467 were identified to be responsible for the bond selectivity of PGUS-E, and further saturation mutagenesis was conducted. Two positive mutants R329K and T369V were obtained by a combined selection technique of thin-layer chromatography and high performance liquid chromatography. Compared to PGUS-E, the bond selectivity of mutants R329K and T369V increased by 26.9% and 34.3%, respectively; whereas the biochemical properties such as pH and temperature profile were unchanged. Nevertheless, the activity was decreased compared to PGUS-E. These results further confirmed that sites R329 and T369 played important roles for the bond selectivity and activity. In summary, this study significantly increased the bond selectivity of PGUS-E by structure guided saturation mutagenesis, providing experimental support for elucidating the relationship between the structure and function of PGUS-E.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Escherichia coli ; metabolism ; Glucuronidase ; chemistry ; Industrial Microbiology ; Mutagenesis ; Recombinant Proteins ; chemistry ; Temperature

OBJECTIVE To investigate the ESBLs-producing isolates and their resistance and genotypes in Escherichia coli and Klebsiella in three hospitals of Wannan area.METHODS The MIC of 13 antimicrobials against 79 strains of E.coli and Klebsiella were determined by 2-fold agar dilution method.The confirmation test was used to detect ESBLs-producing strains according to CLSI 2005.PCR method was used to amplify beta-lactamases of ESBLs-producing isolates by 8 premiers,PCR products were purified and then sequencing was performed.RESULTS The ESBLs-producing rates in E.coli and Klebsiella in three hospitals of Wannan area were 61.5% and 45.0%,respectively.The resistance rates to CRO,CTX,CAZ,CIP and LEV of ESBLs-producing strains were 81.0%,81.0%,47.6%,71.4% and 50.0%,respectively,all isolates were susceptible to meropenem and imipenem.PCR indicated that CTX-M1,CTX-M13,TEM and OXA1 types of beta-lactamases were the four major genotypes of the three hospitals in Wannan area.CONCLUSIONS The isolation rates of ESBLs-producing E.coli and Klebsiella as well as their resistance rates to cephalosporins and quinolones in the three hospitals in Wannan area are quite high.Monitoring ESBLs-producing isolates should be strengthened in clinic and laboratory.

Biopsy ; Biopsy, Needle ; Bone Marrow ; pathology ; Bone Marrow Examination ; methods ; Bone Marrow Neoplasms ; pathology ; secondary ; Breast Neoplasms ; pathology ; Cytological Techniques ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Prostatic Neoplasms ; pathology ; Retrospective Studies ; Stomach Neoplasms ; pathology

OBJECTIVETo analyze the condition of early (≤ 30 d) postoperative pulmonary infection in children after living donor liver transplantation (LDLT).

METHODThe clinical data of 36 cases undergoing LDLT in Children's Hospital of Chongqing Medical University were analyzed retrospectively from June 2006 to December 2009.

RESULTOf 36 cases without preoperative respiratory disease, 17 were boys, 19 were girls. Their age ranged from 2 months to 14 years. Pulmonary infection developed in 24 patients, of whom 4 cases died (17%) and 3 deaths were related to pulmonary infection. Pulmonary infection occurred in 17 of 20 infants (85%) and 10 of 11 cases (91%) with liver function of Child-Pugh grade C. Twenty cases (83%) developed pulmonary infection within first 2 weeks after LDLT. Totally 65 pathogenic strains of microorganisms were isolated, in which Gram-negative bacteria, Gram-positive bacteria and fungi were 46 strains, 5 strains, 14 strains respectively. The most frequently isolated bacteria were Pseudomonas aeruginosa (14 strains), Klebsiella pneumoniae (8 strains) and Acinetobacter baumannii (8 strains). Pseudomonas aeruginosa showed a resistance rate of almost 100% to cotrimoxazole, tetracycline, chloramphenicol, ampicillin, the first, the second and some of the third generation cephalosporins. Klebsiella pneumoniae producing extended spectrum beta-lactamase had a resistance rate of almost 100% to beta-lactams except carbapenems. Acinetobacter baumannii was exquisitely susceptible to carbapenems, but showed a high resistance to penicillins and cephalosporins. Candida albicans, which was the most common fungus, showed a susceptibility rate of 100% to amphotericin B. In the LDLT recipients of pulmonary infection, cytomegalovirus (CMV) infections occurred in 2 patients and Epstein Barr virus (EBV) infection in 1 patient.

CONCLUSIONThe incidence of early postoperative pulmonary infection was high in children undergoing LDLT, especially in infants. And the mortality should not be ignored. The high risk period for infection was within the first 2 weeks after operation. The pathogens were mainly Gram-negative bacteria, which showed high and multidrug resistance.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Antifungal Agents ; therapeutic use ; Bacterial Infections ; drug therapy ; etiology ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Gram-Positive Bacteria ; drug effects ; isolation & purification ; Humans ; Infant ; Liver Transplantation ; Living Donors ; Lung Diseases ; drug therapy ; etiology ; microbiology ; Male ; Postoperative Complications ; drug therapy ; epidemiology ; etiology ; Retrospective Studies ; Risk Factors

OBJECTIVETo observe the effect of Ginkgo Leaves Tablet (GLT) on memory quotient (MQ) of mild cognitive impairment (MCI) patients.

METHODSOne hundred and thirteen patients were randomly assigned to the control group (55 cases) and the treatment group (58 cases). Patients in the control group received dietetic therapy and physical exercises, while those in the treatment group additionally took GLT, 19.2 mg each time, three times daily. The treatment course was 12 months for all. The MQ of all the patients was assessed by WMS-RC before treatment,at 6-month of treatment, and 12-month of treatment.

RESULTSCompared with the control group, the improvement of MQ increased in the treatment group 0.5 and 1 year after treatment (P < 0.05). The clinical efficiency of MQ obviously increased in the treatment group (48.28% and 50.00%), showing statistical difference when compared with the control group (30.91% and 27.27%, P < 0.05, P < 0.01). There was statistical difference in added scores of recognition, regeneration, understanding, and recitation test at 6-month of treatment and 12-month of treatment between the treatment group and the control group (P < 0.05, P < 0.01).

CONCLUSIONGLT was effective in improving MQ of MCI patients, especially in improving recognition, regeneration, understanding, and recitation test.

Aged ; Aged, 80 and over ; Cognitive Dysfunction ; drug therapy ; psychology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; chemistry ; Humans ; Male ; Memory ; Middle Aged ; Phytotherapy

OBJECTIVETo observe the differences in onset time and the overall efficacy of the acupuncture therapy of dredging Governor Vessel and regulating mentality combined with fluoxetine and the fluoxetine for post-stroke depression(PSD).

METHODSSixty-three patients were randomly divided into an acupuncture intervention of dredging Governor Vessel and regulating mentality group(acupuncture and medication group, 33 cases) and a control group (medication group, 30 cases). In the medication group, 20 mg fluoxetine was used by oral administration, once a day at 7:00 in the morning, continuously for 4 weeks. In the acupuncture and medication group, based on the oral administration of fluoxetine, the acupuncture intervention of dredging Governor Vessel and regulating mentality was applied mainly at Baihui(GV 20), Fengfu(GV 16), Shenting(GV 24), Shuigou(GV 26), Dazhui(GV 14) and Shendao(GV 11), once a day, 6 times a week, continuously for 4 weeks. Twenty-four items in Hamilton Depression Scale(HAMD), modified Edinburgh Scandinavia Stroke Scale(MESSS) and activity of daily life scale(ADL, Barthel index, BI) were used before and after 2-week and 4-week treatment. The efficacy was compared between the two groups.

RESULTSAfter 2-week treatment, the HAMD score and the MESSS score in the acupuncture and medication group were obviously decreased and the BI score was apparently increased(all P<0. 01). In the medication group, however, the score of every scale was not statistically different from that before treatment(P'>0. 05). After 4-week treatment, HAMD scores and MESSS scores in the two groups were obviously decreased and the BI scores I were apparently increased(all P<0. 01). After 2-week and 4-week treatment, the HAMD scores and the MESSS scores in the acupuncture and medication group were lower than those in the medication group and the BI scores were higher than those in the medication group(P<0. 01, P<0. 05). The total effective rate of anti-depression (97. 0%, 32/33) the total effective rate of nerve function impairment(90. 9%, 30/33) and the total effective rate of daily life activity(97. 0% 32/33) in the acupuncture and medication group were better than 80. 0% (24/30), 80. 0%(24/30), 83. 3%(25/30) in the medication group(all P<0. 05).

CONCLUSIONThe acupuncture therapy of dredging Governor Vessel and regulating mentality could reduce the onset time of anti-depression medicine treatment of PSD and enhance the overall efficacy. Therefore, it enhances the clinical compliance.

Acupuncture Therapy ; Adult ; Aged ; Antidepressive Agents ; administration & dosage ; Combined Modality Therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Stroke ; complications ; psychology

Objective To investigate the effect of multidisciplinary diagnosis and treatment including ex-utero intrapartum treatment (EXIT) procedure to improve the prenatal survival rate of fetus with neck mass.Methods Multidisciplinary diagnosis and treatment model were carried out in four pregnancy women with fetal neck mass from September 2007 to February 2010.The model included prenatal assessment and monitoring,EXIT procedure during cesarean section,neonatal reassessment and surgical treatment by the cooperation of obstetricians,neonatologists,children surgeons,sonographers and anesthetists.Results All patients underwent cesarean section after 37gestational weeks.Mean delivery time was 37+4 weeks (37-38+3 weeks); mean birth weight was 2972 g (2600-3250 g); mean operation time was 4 min (2-7 min).The gestational age of primary diagnosis of fetal neck mass was 24-34 gestational weeks.After delivery,the size of neck mass was from 3.0 cm × 2.0 cm × 1.0 cm to 6.2 cm× 5.8 cm × 6.8 cm.The tracheal compression and displacement were found by color doppler ultrasound scan and magnetic resonance imaging in all cases.Two of them were completed with polyhydramnios and the others with normal volume of amniotic fluid.EXIT procedure was successfully carried out during cesarean section.Neonatal reassessment showed the trachea of three infants were obviously compressed and lapsed by enhanced CT; the infants relied on mechanical ventilation after birth and underwent operation on day 6 to 8.Tracheal impression was not presented in one infant and trachea cannula was removed on the second day,operation was not performed.All of those infants had good outcomes.Conclusions The multidisciplinary diagnosis and treatment model,including EXIT procedure,is a safe,efficient and feasible strategy,which is necessary for fetus with neck mass.

Objective To measure and define the distribution of left gastric lymph nodes.Meth- ods From Jan.2004 to Apr.2005,silver clips were set around the root of the left gastric artery in 124 pa- tients with esophageal and gastric cardia carcinoma,X-ray films at 0?and 90?simulator gantry in the radio- therapeutic position were taken.Then,the data of the superior,lower,left,right,anterior and posterior bor- der in each patient was recorded.With SAS 8.02 software,data of minimum area which covered the left gas- tric lymph node in different incidences were obtained.Results According to the analysis of Shapiro- Wilk,Kolmogorov-Smimov,Cramer-von Mises and Anderson-Darling tests,each border was of normal distri- bution,with equal frequency in the male and female,despite the actual results in different genders.Pearson Correlation Coefficients analysis did not suggest a significant relationship between the border and height, weight and size of vertebrae,which formed the minimum area covering the left gastric area at frequency of 100%,95%,90% and 85%,which were drawn out through the calculation.Conclusions Aiming at completely identifying the normal distribution of the left gastric lymph node,more patients are required to be in the pool.For the time being,location in the left gastric area can be obtained from details of the results in the present study.

OBJECTIVETo observe the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on biofilm of Staphylococcus epidermidis.

METHODThe serial dilution method was adopted to determine MIC of the combination of sodium houttuyfonate and erythromycin on S. epidermidis; the checkerboard method was used to evaluate the combination of sodium houttuyfonate and erythromycin on suspended bacteria of S. epidermidis; S. epidermidis biofilm was built in vitro, and XTT reduction assay was used to evaluate the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on the adhesion of S. epidermidis and bacterial metabolism inside the biofilm. Microscope was applied to observe the impact the single administration and combination of the two medicines under sub-MIC on biofilm morphology of S. epidermidis.

RESULTThe MIC of sodium houttuyfonate and erythromycin were 62.5, 7.812 5 mg x L(-1), respectively. The combination of 1/8MIC sodium houttuyfonate and 1/2MIC erythronmycin showed a synergistic effect on S. epidermidis. Sodium houttuyfonate, erythromycin and their combination had an inhibitory effect on the adhesion and metabolism of S. epidermidis biofilm bacteria, and made impact on the morphology of S. epidermidis biofilm.

CONCLUSIONThe sub-MIC sodium houttuyfonate and erythromycin have an inhibitory effect on S. epidermidis biofilm. The combination of sodium houttuyfonate and erythromycin shows a synergistic effect in inhibiting suspended bacteria and biofilm of S. epidermidis, particularly in inhibiting the metabolism of S. epidermidis biofilm bacteria and impacting the morphology of biofilm.

Alkanes ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; drug effects ; growth & development ; Dose-Response Relationship, Drug ; Drug Interactions ; Erythromycin ; pharmacology ; Microbial Sensitivity Tests ; Staphylococcus epidermidis ; drug effects ; physiology ; Sulfites ; pharmacology

To investigated Shui nationality folk medicine's awareness to orthopedics & traumatology, the history of orthopedics & traumatology treatment, Shui nationality folk doctors' practicing medicine, heritage, diagnosis and treatment methods and tools, etc, through investigated drug resources category and distribution characteristics of Shui nationality medicine to orthopedics & traumatology treatment, explored and finished Shui nationality medicine orthopedics & traumatology treatment theoretical system. After more than 5 years' exploration and finishing, preliminarily formed the theoretical system framework and medicine application characteristics of Shui nationality medicine treating orthopedics & traumatology. Shui nationality medicine treatment orthopedics & traumatology has distinctive national style, and worthy to further exploration and research.
Bone Diseases ; ethnology ; history ; therapy ; China ; ethnology ; History, 20th Century ; History, 21st Century ; Humans ; Orthopedics ; history ; methods ; Religion and Medicine ; Traumatology ; history ; methods