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 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 .

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.

ObjectiveTo determine the correlation of human leukocyte antigen-G (HLA-G)expression with CMV active infection after kidney transplantation. MethodsA total of 215 first-time kidney transplantation recipients in one transplantation center were divided into CMV ( + ) group and CMV ( - ) group according to whether they had active CMV infection. mhla-g1 expression on leukocytes was analyzed by flow cytometry. The concentrations of soluble HLA-G5 were detected by using ELISA. The sHLA-G5 cutoff levels by ROC curve was employed to predict the active CMV infection. The expression of sHLA-G5 mRNA and protein in leukocytes was analyzed by using RTPCR and Western blotting respectively. Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies of 12 cases. ResultsThe expression of mHLA-G1 in peripheral blood was low in both CMV ( + ) group and CMV ( - ) group. Also when CMV-PP65 was positive, there was no significant change in mHLA-G1. In CMV ( + ) group, the proportion of CD14+ mHLA-G1 +cells[(45. 53 ± 17.32)％]in peripheral blood was increased as compared with that in CMV (-)group[(10. 22 ± 5.78)％]. The expression of sHLA-G5 was increased significantly in CMV ( + )group. The optimal cutoff value of sHLA-G5 predicting the active CMV infection was 202. 9 μg/L,with high diagnostic accuracy. HLA-G was positive in the kidney tissue of 10 patients out of 12 patients with active CMV infection. Both RT-PCR and Western blot analysis showed that sHLA-G5 was significantly higher in CMV ( + ) group than that in CMV ( - ) group. ConclusionROC curve analysis of sHLA-G5 with the cutoff value of 202. 9 μg/L can be used to predict the active CMV infection. The HLA-G levels in peripheral blood were significantly increased and HLA-G expression in the tubular epithelial cells of the graft could be a protection mechanism of the kidney function.

ObjectiveTo evaluate the relationship between the proliferation of parathyroid cell in rabbit with primary hyperparathyroidism (PHPT) and the bone mineral density (BMD). MethodsEighty adult Chinese rabbits were randomly and equally divided into two groups. The contrast group was fed with normal diet ( Ca ∶ P, 1.0 ∶ 0. 7 ) and the experimental group was fed with high phosphate diet ( Ca ∶ P,1.0∶7.0) to establish the animal model of PHPT. At 3, 4, 5, and 6 months after the diet, bone mineral density of the rabbits was measured by the quantity CT (QCT). Then, the parathyroid and bone of the rabbits were removed for pathological examination. The number of parathyroid cell in PHPT was calculated.Proliferation was determined by immunohistochemistry of proliferation cell nuclear antigen ( PCNA ) and Bcl-2. The t test and Logistic regression was used to analyze the difference of data of two groups. ResultThe number of parathyroid cell in PHPT group was 1.61 times than that in the contrast group[ (673 ± 151 ) HP,(418 ± 25 ) HP,P ＜0. 01]. The rate of PCNA positive-cell was significantly increased in PHPT group than that in contrast group [(50.52 ± 11.62)％o, (26.70 ± 2. 78 )％, P ＜ 0.01], and so was Bcl-2[ (460. 37 ± 190. 05 )‰, (67. 02 ±:4. 38 )％‰,P ＜0. 05]. The value of BMD was significantly decreased in PHPT group than that in contrast group [ ( 152. 5 ± 34. 3 ), ( 188.6 ± 12. 2 ) g/cm3, P ＜ 0. 05]. There was a negative correlation between BMD and PCNA (r ＝ -0. 749, P ＜ 0. 05 ) and between BMD and Bcl-2 (r ＝-0.800, P ＜ 0. 05 ) in PHPT group. ConclusionThe BMD of PHPT is related to the parathyroid cells proliferation which provide a reliable method for early diagnosis of PHPT.

Objective To study the correlation of HLA-G levels with acute rejection and CMV active infection post-kidney transplantation.Methods A total of 132 initial kidney transplantation recipients were divided into kidney function stable group (F),acute rejection group (AR),CMV group according to whether they had active CMV infection and acute rejection.Forty-one healthy donors served as control group (H).HLA-G levels and mRNA expression were analyzed by using flow cytometry,ELISA,RT-PCR and Western blotting.Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies.Results The expression levels of mHLA-G1 were low in all 4 groups pre-transplantation.Only CMV group had significantly more CD14+ mHLA-G1+ cells post-transplantation (P＜0.05).sHLA-G5 levels were higher in F group than in H group (P＜0.05),but there was no significant difference among other groups pre-transplantation (P＞0.05).sHLA-G5 levels were increased significantly in CMV group as compared with F group (P＜0.05),and those in F group were higher than in H and AR groups (P＜0.05).Renal tissue biopsies from 21 renal transplantation recipients with AR indicated that HLA-G5 was expressed negatively in 17 patients,positively in 3 patients and 1 weakly positively.HLA-G was positive in the kidney tissue of 9 patients out of 9 patients with active CMV infection.In total 132 recipients,AR incidence was significantly lower in CMV ( + ) group (7.1 ％,2/28) than that in CMV ( - ) group (24.0 ％,25/104).Conclusion The sHLA-G5 may contribute to predict AR and CMV active infection; AR and CMV active infection may be correlation with immune balance in kidney transplantation recipients.

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.