OBJECTIVE To investigate the drug resistance and the status of producing ESBLs and AmpC beta-lactamases of Klebsiella pneumoniae isolates from 2006 to 2007 in local district.METHODS From 2006 to 2007,110 strains of K.pneumoniae insensitive to cefoxitin were collected.The sensitiveness to 16 antibiotics were tested by K-B method and microdilution method.Genes of TEM,SHV,GES,PER,CTX-M-1,CTX-M-2,CTX-M-3,DHA and MIR-1/ACT-1 were tested by PCR.The gene transfer was detected by conjugation test.RESULTS The resistance rate of 110 K.pneumoniae strains to meropenem,imipenem,piperacillin/tazobactam,cefoperazone/sulbactam,ceftazidime and cefepime was 0-49.9%.The resistance rate to other antibiotics was 80-100%.And ESBLs production was the main result.Genes of ESBLs were CTX-M and SHV.Genes of AmpC beta-lactamases were ACT and DHA.They all could transfer the drug-resistance from plasmid to receptor bacteria.CONCLUSIONS Co-existing of ESBLs and AmpC beta-lactamases is the main reason of multi-drug resistance that K.pneumoniae.Transferring of drug-resistance gene leads to the spreading of drug-resistance.The drug-resistance rate of K.pneumoniae decreased during the last two years.

OBJECTIVE To investigate the incidence rate of Pseudomonas aeruginosa whose ceftazidime resistance was induced by imipenem and the relationship of excretion pump with sensibility of imipenem and ceftazidime.METHODS The sensibility to 15 antibacterials and ceftazidime resistance induced by imipenem were detected by slip diffusion method,detect these bacteriumas′s MIC to Imipenem and Ceftazidime before and after excretion pump inhibitor hydroxide radical cyanogen chlorine phenylhydrazone(CCCP) were added by agar dilution,detect Ceftazidime′s MIC value which were added different concentration imipenem by agar dilution.RESULTS From 325 strains 116 strains(35.7%) were imipenem induced ceftazidime-resistant drug fast,from them 80.2% were imipenem-resistant and ceftazidime-sensitive,19.8% were imipenem and ceftazidime both sensitive.Sensibility to imipenem and ceftazidime was no marked change before and after CCCP added,MIC value of ceftazidime was step up 4-12 times by imipenem.CONCLUSIONS The incidence rate of ceftazidime-resistant P.aeruginosa induced by imipenem is high,clinical microbiological laboratory should evolve D-test to guide clinician use antibacterials more reasonable.

Objective To study the feasibility of renal hypothermia achieved by retrograde icecold saline infusion with perfusion pump. Methods Twenty-one patients who received the open radical nephreetomy were divided into three groups,A group with ice-slush renal hypothermia,B group with gravitational retrograde ice-cold infusion,C group with retrograde ice-cold infusion by perfusion pump.After the kidney was dissected and the ureter was divided,the renal vasculars were clamped.The kidney was cooled by three methods respectively.The temperature of renal parenchyma was monitored for 15 min. Results Fifteen minutes later,the temperature of renal parenchyma of A group declined from 34.4℃ to 5.4℃,B group from 34.8℃ tO 23.8℃,C group from 35.1℃ to 22.3℃.Coneiusions Renal hypothermia can be achieved by retrograde cold saline infusion and the perfusion pump may accelerate the speed of cooling.

Objective To investigate the regulatory effects of umbilical cord-derived mesenchymal stem cells (UC -MSCs) on Th17 cells and related cytokines in patients with systemic lupus erythematosus (SLE). Methods Human UC-MSCs were isolated and expanded and infused into fourteen SLE patients. Clinical changes were evaluated before and after transplantation by SLE disease activity index (SLEDAI), 24-hour urine protein, serum albumin and complement C3. The percentages of CD3 +CD8-IL17A + T cells in peripheral blood were detected by flow cytometry. Concentrations of plasma IL-6, TGF-β, IL-17A, IL-22were determined by enzyme-linked immunosorbent assay (ELISA). UC-MSCs and peripheral blood mononindependent samples t-test. Results SLEDAI scores decreased significantly at 3 month (7.8±1.2, t=2.19) and 6 month (6.9±0.9, t=4.2) after UC-MSCs transplantation than pre-transplantation level (10.4±0.9, P< 0.05). Twenty-four-hour proteinuria decreased significantly 6 months after MSCs infusion [(1489±260) mg vs (2454±322) mg, t=2.6, P<0.05]. Meanwhile, serum albumin and complement C3 levels had increased significantly since 1 month after transplantation (P<0.05). The percentages of peripheral blood CD3+CD8-IL17A+T cells decreased obviously in 1 week, 1 month and 6 months after UC-MSCs transplantation (all P<0.05). The coculture of UC-MSCs with PBMC from SLE patients resulted in a statistically significant reduction of CD3+CD8-IL17A+T cells percentage in PBMC (P<0.05), but was not in a dose dependent manner. No change of plasma IL+6, TGF-β, IL-17A and IL-22 levels was observed after UC-MSCs transplantation (P>0.05).Conclusion UC-MSCs transplantation down-regulates the percentages of CD3+CD8-IL17A+T cells in SLE patients, which may be one of the mechanisms for its therapeutic effect in refractory SLE.

Objective To explore the potential predominance and value of percutaneous hepatocholangiostomy (PCH) in treatment of recurrence hepatobiliary stones (RHS). Methods Seventeen cases with RHS were treated by PCH from February 2001 to October 2005, which was an improved technology of percutaneous transhepatic cholangioscopy (PTCS) and made reference to the methods of percutaneous nephrostomy (PCN). Results Seventeen cases were successfully treated. The average blood loss was 40 ml (ranging from 15 to 100 ml), and stones were removed completely in 88.2% (15/17). The average hospital stay was 14 (ranging from 10 to 59) days. No one required postoperative analgesic. No postoperative bleeding and biliary leakage were found. Conclusions PCH has significant advantages of minimal invasion, little blood loss, less pain, less complications and quick recovery in the treatment of RHS.

Objective To study the relationship of Wnt receptor signaling pathway and severe retinopathy of prematurity (ROP).Method From January 2011 to June 2015,preterm infants with severe ROP admitted to the NICU of our hospital were enrolled prospectively.Preterm infants with similar gestational age,gender,and age (in days) admitted to our hospital during the same period were selected as the control group.FZD4,LRP5,and ND gene mutations in Wnt receptor signaling pathway were examined.Result A total of 61 Chinese preterm infants were screened for these three candidate genes of Wnt receptor signaling pathway,32 in ROP group and 29 in control group.ND and FZD4 gene mutations were not found among all cases.Eight types of LRP5 mutations were found in 26 cases of ROP group,including 7 cases of Exon18 missense mutation [c.3989C ＞ T;p.Ala1330Val (rs3736228)],5 cases of Exon8 synonymous mutation (c.1647T ＞ C;p.Phe549Phe),5 cases of Exon6 intronic mutation [c.1412 + 8G ＞ A (rs4988319)],3 cases of Exon2 missense mutation [c.266A ＞ G;p.Gln89Arg (rs41494349)],2 cases of Exon21 intronic mutation [c.4349-17C ＞ T (rs372086596)],2 cases of Exon19 synonymous mutation (c.4089C ＞ T;p.Asp 1363 Asp),one case of Exon9 synonymous mutation (c.1 932G ＞ A;p.Glu644Glu),and one case of Exon16 missense mutation (c.3580C ＞T;p.Arg1194Cys).Three types of LRP5 mutations were found in 6 cases of the control group,including 4 cases of Exon8 synonymous mutation,one case of Exon19 synonymous mutation,and one case of Exon9 synonymous mutation.The positive rates of Exonl8 missense mutation and Exon6 intronic mutation in severe ROP group were significantly higher than the control group (P ＜ 0.05).Conclusion LRP5 gene mutations in Wnt receptor signaling pathway may be associated with the occurrence of severe ROP.

Objective To improve the arthroscopic posterior cruciate ligament (PCL) reconstruction using tibial Inlay technique. Methods The special arthroscopic device and related fixation technique were designed. Five cadaveric knees were used to simulate the process of arthroscopic posterior cruciate ligament reconstruction using tibial Inlay technique. The knees were cut open to observe whether the outlet of the tibial tunnel shape and location met the design requirements. Thirty normal MRI films were measured to identify tunnel angle and localizer angle. Results The inner outlet of tunnel was conical shape(14 mm×7 mm×15 mm) and the outer outlet was cylinder-shaped (a diameter of 7 mm). The tibial drill was designed into a split structure and could be assembled in vitro. According to the data obtained from MRI films, the angle between the plane of posterior cruciate ligament and horizontal place was 36°-47°, and the localizer was fixed at 50°.The achilles tendon was used as implant and the allogft bones were designed into conical shape to fit the inner outlet of tunnel. The other end of implant to the proximal tibia was fixed with button plate. All reconstruction operations were performed under arthroscopy. The outcomes of procedure were satisfactory. There were no vascular or peripheral nerve injuries in the cadaveric knees The tunnel position was accurate and the shape of tunnel had met the design requirements. Conclusion Our results imply that improved arthroscopic of posterior cruciate ligament using tibial Inlay technique is simple, accurate, rapid and stable fixation.

The aim of this study is to investigate the effect and mechanism of angiotensin (Ang) II on E-selectin and vascular cell adhesion molecule-1 (VCAM-1) expression in rat brain microvascular endothelial cells (BMEC) and evaluate the effect of compound EXP-2528, a novel Ang Ⅱ type 1 (AT1) receptor antagonist. The experiment was performed in cultured BMEC of rat. The mRNA and protein expression of E-selectin and VCAM-1 in BMEC was analyzed by RT-PCR and Western blotting, respectively. The results showed that the mRNA and protein expression of E-selectin and VCAM-1 in BMEC were significantly upregulated by 4 h or 18 h exposure to 1×10-7 mol·L-1 Ang Ⅱ. These effects were abolished by pretreatment with the selective AT1 receptor antagonists losartan and compound EXP-2528, but not with the AT2 selective antagonist PD123319. Combining losartan with PD123319 also significantly inhibited Ang Ⅱ-induced E-selectin and VCAM-1 expression in BMEC, but there was no significant difference compared with losartan group. These findings indicated that Ang Ⅱ upregulated E-selectin and VCAM-1 in BMEC by activating AT1 receptor and then involved in the development of cerebrovascular disease.

Objective To investigate the effect and mechanism of murine bone marrow mesenchymal stem cells(BM-MSCs)transplantation on B-cell activating factor(BAFF)expression and B cell activation of MRL/Ipr mice.Methods Eighteen female MRL/Ipr mice were divided into the treatment group and the control group.Five female BAL B/C mice were used as negative controls.At the age of 18 weeks,the treatment group was transplanted with 1×10~6 murine BM-MSCs through vena caudalis,the control group was treated with 0.5ml sodium chloride.Enzyme linked immunoserbent assay(ELISA)was used to measure the level of the BAFF,IFN-γ,IL-2 and IL-10 in the serum.The percentage and numbers of Marginal zone,T1 and T2 B cells in spleen were detected by flow cytometry.Results①Eight weeks after transplantation,the level of BAFF [(32±14)ng/ml]in serum of the treatment group decreased significantly than the control group[(47±13)ng/ml](P＜0.05)as well as the level of serum IL- 10,IFN-γ and IL-2 levels[(19±7)vs(40±13)pg/ml](P＜0.01)[(25±20)pg/ml vs(38±25)pg/ml][(73±10)pg/ml vs(80±15)pg/ml].② Eight weeks after trans-plantation,the mice in the treatment group had lower percentages of marginal zone B cells[(15±4)% vs (21±5)%],and the numbers of marginal zone B cells were significantly decreased in the treatment group as compared with the control group[(9±6)×10~6 vs(19±10)×10~6,P＜0.05].③ Eight weeks after transplantation,the mice in the treatment group had lower percentages of T1 and T2 B cells[(3.4±2.1)% vs(7.3±4.0)%][(2.6+1.4)%vs(4.8±2.7)%],and the numbers of T1[(2.7±1.7)×10~6 vs(5.1±2.0)×10~6,P＜0.05]and T2 B cells[(2.0±1.2)×10~6 vs(3.7±1.7)×10~6,P＜0.05]were both significantly decreased in the treatment group as compared with the control group.Conclusion BM-MSCs transplantation decreases the expression of BAFF in association with the diminished production of the pathogenic cytokines IFN-γ and IL-10.Inhibition of BAFF also results in decreased numbers of T1 and T2 B cells and MZ B cells.

Objective To evaluate the efficacy and safety of 650 nm,810 nm diode laser on the treatment of moderate to severe acne.Methods A total of 87 patients with moderate to severe acne were divided into diode laser group (group A) and medication group (group B).In group A,the patients were irradiated with 650,810 nm diode laser at the fluence of 400-450 mW distance of 2-3 cm.In group B,the patients were treated with roxithromycin combined with vitamin B6 and gel lincomycini.Clinical follow-up was performed every week for 4 months.All the adverse effects were recorded.Results There was no significantly statistical difference in the cure rate between the two groups (group A versus group B at 2 weeks:65.0 ％ versus 42.6 ％,P＞0.05; at 4 weeks:92.5 ％ versus 76.5 ％,P＞0.05).However,the recurrence rate of group A was much lower than that of group B (30.0 ％ versus 61.7 ％,P＜0.01).There were no severe adverse effects in both groups during the period of the follow-up.Conclusions Compared to the medication,650,810 nm diode laser appears to be associated with lower recurrence rate in the treatment of moderate to severe acne.