OBJECTIVE To study the differences of quinolones-resistant determining region(QRDR) in DNA gyrase A(gyrA) subunit between drug-resistant Klebsiella pneumoniae induced by fluoroquinolone in vitro and strains isolated from clinical isolates.Furthermore,to reveal the relation between different QRDR gene mutations and drug-resistance. METHODS Ten sensitive K.pneumoniae strains were selected and induced into drug-resistant strains by ciprofloxacin.The QRDR in gyrA by polymerase chain reaction(PCR) was amplified and compared their DNA sequences with clinically isolated quinolone-resistant K.pneumoniae. RESULTS We had found several gene mutations of QRDR in fluoroquinolone-resistant K.pneumoniae induced by ciprofloxacin,including Ser 83(TCC)→Phe(TTC) and Ile(ATC);and Gln106(CAG)→Leu(CTG).And the alteration of Gln106(CAG)→Leu(CTG) was a new discovery. CONCLUSIONS There are gene mutations of QRDR in fluoroquinolone-resistant K.pneumoniae induced by ciprofloxacin.The mutation of Ser 83(TCC) and Gln106(CAG) is probably the molecular basis of inducing drug-resistant strains.

Objectives To investigate the methods of protecting external branch of the superior laryngeal nerve (EBSLN)in carotid endarterectomy and to observe the effect of using these methods in clinical surgery. Methods EBSLN (20 sides)of 10 heads of corpse were studied by using microanatomy from January 2013 to December 2013. The occurrence probability of EBSLN on the lower edge of posterior belly of digastric muscle,medial edge of external carotid artery and upper edge of superior thyroid artery in anatomy triangle was analyzed. The distances from the midpoint of the EBSLN to carotid bifurcation, mandibular angle and mastoid tip were measured. Sixty-five patients with carotid endarterectomy in Tianjin Huanhu Hospital from December 2013 to November 2014 were treated with the protective methods of the relevant EBSLN by using anatomy triangle as a mark. Whether the patients had injury symptoms of EBSLN were followed up after procedure. Results (1)The occurrence probability of 20-side EBSLN in anatomy triangle was 95%(19 sides). The midpoint of EBSLN in the anatomy triangle at the posterior mandibular angle was median 0. 34 (-1. 62 to 2. 43)cm,at the inferior mandibular angle was 1. 28 (-1. 33 to 3. 42) cm,at anterior mastoid tip was 2. 84 (0. 51 to 5. 14)cm,at inferior was 4. 51 (2. 82 to 6. 39)cm,and at anterior superior of the carotid bifurcation was 1. 64 (0. 57 to 3. 78)cm. (2)65 patients who underwent carotid endarterectomy used the protective methods of intraoperative EBSLN. There was no manifestation of EBSLN injury at 3 weeks to 9 months after procedure. Conclusion In carotid endarterectomy,taking an anatomic triangle as a symbol,it is no more than 2 cm of the anterior superior of carotid bifurcation during the separation process. As for the patients with higher or lower position of carotid bifurcation,in the range of crossing rear mandibular angle 0. 50 cm or below the mastoid tip 4. 50 cm for arterial separation should be avoided,and this can effectively protect EBSLN.

Objective To evaluate the protective effect of therapeutic hypercapnia on the lung during one-lung ventilation (OLV) in the patients undergoing pulmonary lobectomy.Methods Fifty patients of both sexes,aged 20-60 yr,with body mass index 18-30 kg/m2,of American Society of Anesthesiologists physical status Ⅱ,scheduled for elective pulmonary lobectomy,were randomly divided into 2 groups (n=25 each) using a random number table:control group (group C) and therapeutic hypercapnia group (group H).After induction of general anesthesia,the patients were endotracheally intubated and mechanically ventilated in volume-controlled mode.The ventilator settings were adjusted during two-lung ventilation to maintain the end-tidal pressure of carbon dioxide (PETCO2) at 25-35 mmHg.Group H inhaled the mixture of CO2 (3％-6％) and O2 (70％-82％) during OLV to maintain PETCO2 at 50-60 mmHg.Group C inhaled O2 (70％-88％) during OLV to maintain PETCO2 at 25-35 mmHg.Anesthesia was maintained with inhalation of sevoflurane and intravenous infusion of remifentanil.Immediately before OLV and at 30 min after restoration of two-lung ventilation,the airway peak pressure,airway plateau pressure and lung compliance were recorded,arterial blood samples were collected for blood gas analysis,and broncho-alveolar lavage fluid (BALF) from the collapsed lung and venous blood samples were collected for determination of tumor necrosis factor-alpha (TNF-α),interleukin-1beta (IL-1β),IL-6,IL-8 and IL-10 concentrations in BALF and serum by enzyme-linked immunosorbent assay.Oxygenation index was calculated.Results Compared with group C,the airway peak pressure and airway plateau pressure were significantly decreased,the lung compliance was significantly increased,the concentrations of TNF-α,IL-1β,IL-6 and IL-8 in BALF were significantly decreased,and the concentrations of IL-10 in BALF were significantly increased at 30 min after restoration of two-lung ventilation (P＜0.05),and no significant change was found in the oxygenation index and concentrations of inflammatory factors in serum in group H (P ＞ 0.05).Conclusion Therapeutic hypercapnia can improve pneumodynamics and attenuate inflammatory responses,and has no significant difference clinically in improving oxygenation during OLV in the patients undergoing pulmonary lobectomy.

Objective To explore short-term effect of tonsillotomy on children′s obstructive sleep apnea hypopnea syndrome (OSAHS) with tonsil hypertrophy by Ellman double-frequency radio-frequency machine .Methods Totally 30 cases children′s OS-AHS with tonsil hypertrophy were had tonsillotomy by Ellman double-frequency radio-frequency machine .They were compared with 30 cases children′s OSAHS with tonsil hypertrophy by traditional tonsillectomy .Short-term effect of tonsillotomy were ana-lyzed by wound bleeding ,operation time ,postoperative pain ,difficulty eating ,PSG testing ,immune function ,growth and develop-ment .Results Postoperative symptom in 1 weeks and 6 months ,compared with the traditional tonsillectomy ,30 cases of OSAHS patients were found that there were less bleeding ,shorter operative time ,mild postoperative pain ,no difficulty in feeding ,improving sleep-disordered breathing ,no decline in immune function ,good growth and development .Conclusion Short-term effect of tonsillot-omy on children′s OSAHS with tonsil hypertrophy is good by Ellman double-frequency radio-frequency machine ,it is a minimally invasive ,safe and effective surgical method children OSA HS .

Objective: To give the living-donor and recipient medical evaluation and moral evaluation pretransplant by Academic and Ethics Committee for Organ Transplantation (AECOT), ensure the transplantation proceed smoothly and the feasibility for moral principles. Methods:AECOT was formed and gave the living-donor and recipient medical evaluation, additional moral evaluation to their family, evaluated the possibility for transplantation, ensured the long-dated safety for living-donor and recognition for ethics, ensured the transplantation proceed successfully. Results: 18 cases were evaluated by AECOT, two cases had the operation refused because of the II-diabetes for one living-donor and the other living-donor refused evaluation, one case had the operation delaied because of the overweight for the living-donor. The rest 15 cases were accepted operation smoothly,both donor and recipient recovered well post transplant. Conclusion:Academic and Ethics Committee for Organ Transplantation(AECOT) is a essential agency for Living Donor Kidney Transplantation.

Objective To evaluate the mid-term tollow-up results of cervical artificial disc replacement (CADR) for cervical degenerative disc disease,and to explore whether it can reduce the occurrence of adjacent segment degeneration (ASD).Methods A prospective comparative study of 93 patients who underwent CADR or anterior cervical decompression and fusion (ACDF) for cervical degenerative disc disease were conducted.All patients were followed up for more than 6 years.The Japanese Orthopaedic Association (JOA) score,neck disability index (NDI),Odom's scale,X-rays and magnetic resonance imaging (MRI) were used to evaluate the clinical and radiologic results.Results Twenty eight patients who underwent CADR and 35 patients who underwent ACDF had complete follow-up data.At final follow-up,the JOA score and NDI improved significantly in both groups.Between the two groups,there was no significant difference in terms of JOA score,NDI and Odom's scale.The sagittal alignment was well maintained in both groups.The total cervical spine range of motion (ROM) had no significant change for the CADR group,whereas,it significantly decreased for the ACDF group.The ROM at the replacement level of CADR patients decreased from 9.5° ± 3.7° before operation to 7.0° ± 3.0° 3 months after operation,and it was maintained to 6.6° ± 4.1° at final follow-up without significant decrease.Lateral radiographs and T2-weighted MRI showed the incidence of ASD in CADR group was significantly lower than that in ACDF group.Conclusion The six-year follow-up results of CADR are basically satisfactory.Compared with ACDF,it could better preserve physiological motion and biomechanics of cervical spine,and reduce the incidence of ASD.

High-competence specialists are elites among health professionals.By means of the authoritative training system,the hospital took such measures as,strengthened management,rotary training without fixed posts,and cross training.Efforts of 8 years have helped the hospital to establish a high-competence specialists team,a good academic atmosphere,and enhanced capacity of disciplines. Key experiences of the hospital in this process include well-identified objectives,well-designed training programs, strict supervision of training process, and establishment of long-term management mechanism.

Objective: To investigate the effect of losartan on angiotensin II (Ang II) expression and myocardial remodeling in myocardial infarction (MI) rats’ model.
Methods: A total of 32 SD male rats were divided into 4 groups, Sham operation group, MI group, MI with losartan 10mg/(kg·d) group and MI with losartan 20mg/(kg·d). n=8 in each group. MI model was established and the electrocardiogram changes before and after MI were recorded, hemodynamic indexes were detected at 4 weeks after MI, pathological changes of myocardial tissue were examined by HE staining. The myocardial mRNA and protein expressions of ACE2 and Ang II were detected by RT-PCR and Western Blot analysis.
Results: Compared with Sham operation group, MI group showed increased LVMI and decreased LVEF P<0.05;the above changes were getting better in both MI with losartan groups in a dose-dependent manner. The pathological examination presented that MI group had myocardial cell swelling, fracture, hyperplasia and inflammatory cell infiltration, those damages were less in MI with losartan groups in a dose-dependent manner, Sham operation group had no pathological changes. Compared with Sham operation group, the mRNA and protein expressions of Ang II were obviously higher in MI group, P<0.05 and the expressions were decreased in MI with losartan groups in a dose-dependent manner;the mRNA and protein expressions of ACE2 were slightly increased in MI group and the expressions were further increased in MI with losartan groups in a dose-dependent manner.
Conclusion: Losartan could increase ACE2 expression and therefore, inhibit Ang II expression and improve the ventricular remodeling in MI rats’ model.

Matrix_assisted laser desorption ionization_time of flight tandem mass spectrometry ( MALDI_TOF/TOF MS) and electrospray ionization_quadrupole_time of flight mass spectrometry ( ESI_Q_TOF MS) were used to confirm the structure of cyclic lipopeptide daptomycin fastly. First, the relative molecular weight 1916. 7107 of daptomycin was measured by ESI with error 0. 0007. The sample’s doubly charged peak m/z 809. 848 was selected as precursor ion for ESI_MS/MS analysis, and the exocyclic amino acid sequence C9 H19 CO_Trp_Asn_Asp was successfully matched. Second, the experimental conditions of cleaving daptomycin by lithium hydroxide ( LiOH) were optimized and the ring_opened process was monitored by MALDI_TOF/TOF MS. After obtaining ring_opened product with purity of above 95%, the MS/MS measurements by MALDI and ESI were carried out. The b+and y+of ring_opened product were completely matched, which confirmed the amino acid sequence of daptomycin. Finally, ESI_MS/MS conditions of ring_opened product were further optimized to obtain more low mass fragment ions for analyzing the structure of fatty acid chain and the cleavage pattern of fat chain in mass spectrometry was proposed. The method was fast, convenient, accurate and reliable for identifying cyclic lipopeptide compounds.

A novel series of fingolimod analogues containing diphenyl ether moiety were designed and synthesized based on the modification of immunosuppressive agent fingolimod used in the treatment of multiple sclerosis. Compounds were evaluated in vivo for lymphopenic activity and heart rate affection. Most compounds showed moderate lymphopenic activity. It is worth noting that compounds 6c, 6d and 14c-14e showed considerable immunosuppressive activities comparable to fingolimod. And compound 14e had no effect on heart rate.