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.

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 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 .

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 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.

Objective To measure the cytokines levels in peripheral blood from kidney transplantation recipients by using cytometric bead array and to analyze their change and the clinical significance in pre- and post- kidney transplantation, inducting with basiliximab and graft rejection. Methods A total of 72 renal transplantation recipients were divided into two groups, kidney function stable group(n =53) and acute rejection group (n = 19). And they were also grouped by induction with basiliximab or not,32 in basiliximab group and 40 in without basilixmab group. The levels of IFN-γ, TNF-α, IL-10, IL-5,IL-4, IL-2 were measured by cytometric bead array in peripheral blood of 72 kidney transplantation recipients and 30 healthy donors at differential time. The data was analyzed according to the following grouping:donors and recipients, kidney function stable group and acute rejection group post transplantation and with or without basiliximab group. Results The levels of TNF-α, IL-10, IL-5, IL-4, IL-2 in recipients before transplantation were ( 1.65 ±0. 10) ,(2. 55 ±0. 19) ,( 1.88 ±0. 14) ,(1.85 ±0. 12) ,(2. 12 ±0. 09) ng/L,respectively. While they were (3.04 ±0. 17), (3.33 ±0. 26), (4.03 ±0.25), (2.73 ±0. 16), (4.03 ±0. 26) ng/L respectively in healthy donors. There was statistical significance between the two groups ( t =6. 890, 2. 375, 7. 851,3.955,7.153, P<0. 01, <0. 05, <0.01, <0.01, <0.01). While the level of IFN-γ in recipients before transplantation was (2. 50 ±0. 18) ng/L,compared with (3. 00 ±0. 24) ng/L in healthy donors. There was no statistical significance between the two groups( t = 1. 625, P > 0. 05 ). The levels of IFN-γ and IL-10 in kidney function stable group were (2. 71 ± 0. 11 ) ng/L and (3.91 ± 0. 52) ng/L,while they were ( 3.30 ± 0. 36 ) ng/L and ( 12. 01 ± 5.35 ) ng/L in acute rejection group. There were statistical dirrerences between the two groups ( t = 5. 061, 11. 465, P < 0. 01, < 0. 05 ). Before induction with basiliximab, the levels of IFN-γ, TNF-α, IL-10 in recipients were (2.90 ±0. 21 ), ( 1.67 ±0. 12),(2. 45 ± 0. 16) ng/L respectively. But they were ( 2. 78 ± 0. 17 ), ( 1.58 ± 0. 07 ), ( 2. 77 ± 0. 24 ) ng/L respectively after induction with basiliximab, which showed significantly different ( t = 5. 605, 6.011,4. 126, P <0. 01, <0. 01, <0. 05). Four weeks after kidney transplantation in recipients with basiliximab,the levels of IFN-γ, IL-10, IL-4 were (2. 90 ± 0. 31 ), (9. 08 ± 0. 16), (2. 73 ± 0. 11 ) ng/L. While they were (3.28 ±0. 11 ), (4. 17 ±0. 21 ), (2. 11 ±0. 20) ng/L respectively in recipients without basiliximab induction, which were significantly different from those with basiliximab induction (t = 4. 268,4. 263,3.762, P <0. 01, <0. 01, < 0. 05 ). Conclusions Six kinds of cytokines can be measured by cytometric bead array simultaneously and accurately. The data suggests that the detection of multiple cytokines in kidney transplantation recipients by cytometric bead array can provide more guidance for clinical diagnosis and therapy.

Objective To discuss the significance of near infrared spectroscopy (NIRS) in evaluation of intrapartum hypoxic-ischemic cerebral injury, and to provide a method to evaluate neonatal brain damage objectively and quantitatively. Methods A total of 63 neonates with fetal distress were divided into hypoxic-ischemic encephalopathy(HIE) group and non-HIE group. Thirtyfive newborns with no fetal distress were chosen as controls. Using NIRS, the brain regional oxygen saturation(rSO2) in these neonates were measured. Evaluation of brain rSO2 in the diagnosis of HIE was analyzed with receiver operating characteristic (ROC) curve. Results At the time of fetal head visible on vulval gapping and 5 min after birth, the HIE group showed decreased brain rSO2[(36. 6±5.0)% and (52. 0±4. 2)%], comparing with control group[(45. 9±4. 6)% and (59. 6±4. 4)%]and non-HIE group[(44.1±3.1) % and (57. 6±3. 5) %](P<0. 01) . The brain rSO2 was positively correlated with the pH and oxygen saturation of umbilical artery blood in all groups (P<0. 01). When the cut-off value of brain rSO2 was <39. 5% at fetal head visible on vulval gapping, the sensitivity and specificity of assessing HIE were 67% and 93%, respectively, while 70% and 86% when the cut-off value was <53. 5% at 5 min after birth. Conclusions The brain rSO2 obtained by NIRS could be used to evaluate brain oxygenation, and may be useful in predicting HIE in neonates with fetal distress.

Objective To analyze C4d deposition in the patients with late acute renal allograft rejection,and explore the role of C4d in grafts survival and grafts loss. Methods Thirty-six patients clinical and pathologically diagnosed as having acute rejection more than one year post-transplant were selected. C4d was detected by immunohistochemistry in renal allograft biopsies. The effect of C4d deposition on long-term graft survival was studied. Results Among 36 recipients with late acute renal allograft rejection, 16 cases were positive for C4d (44.4 %) and 20 negative for C4d (55.6 %). Five cases experienced graft loss in C4d positive group (31.3 %), while 6 cases in C4d negative group (30.0%). There was no significant difference in the graft loss rate between C4d-positive group and C4d-negative group. Log-Rank test demonstrated there was no significant difference in graft survival between C4d-positive group and C4d-negative group. The count of the interstitial infiltrated eosinophils in renal allograft was (9.4 + 4.5) and (2.6 + 1.8) respectively in the C4d-positive group and C4dnegative group (P＜0.05). Conclusion C4d deposition in peritubular capillary of the recipients with late acute renal allograft rejection might not be a prognostic marker for graft outcome.

ObjectiveTo explore the bone anabolic effects after a single local injection of simvastatin into femoral cavities of osteoporotic rats.MethodsThirty-six female SD rats(3 months old,body weight 250-300 g) were ovariectomized(OVX) and low-calcium-diet fed for 3 months,OVX rats were randomized into 3 groups(n=12).Left femurs of group A,B and C were injected with 0,5 and 10 mg simvastatin,respectively.Half of the rats in each group were randomly euthanized separately 1 and 5 months after simvastatin injection.Left femurs were taken out for bone mineral density (BMD) assessment with dual energy X-ray absorptiometry,bone histomorphometic changes were analysized by Micro-CT,and two kinds of biomechanical tests were used to evaluate the osteogenic effects.ResultsOne and five months after injection,BMD in mid-diaphysis significantly increased in simvastatin-injected groups compared to the control group.For Micro-CT analysis,significant increase in total bone volume/total tissue volume,cortical wall thickness,trabecular thickness,trabecular number,and a significant decrease in trabecular spacing were observed in simvastatin-injected groups compared to the control group.For both biomechanics (the three-pointbreaking test of condyles and axial compressive testing of proximal femur),the values were significantly higher in simvastatin-injected groups than the control group.ConclusionLocal simvastatin treatment showed a positive effect on improving mechanical strength,structure of osteopenic femurs and BMD.Our findings may provide a new strategy for the prevention and treatment of osteoporosis,especially for osteoporotic fractures.