Aim To study gyrA and parC mutations of clinical Pseudomonas aeruginosa strains. Methods MIC values of 55 clinical P.aeruginosa isolates were determined by agar dilution test and 1 sensitive strain and 8 resistant strains were selected with standard sensitive strain ATCC27853 as control, the quinolone determining region (QRDR) of the gyrA and parC genes were amplified by PCR, the lengths of PCR products were 351 bp and 397 bp. The gyrA PCR products(351 bp) were digested with enzyme sacⅡ. The gyrA and parC gene were sequenced. Results In this study, gyrA genes of all resistant strains had an ACC to ATC mutation in codon 83, leading to the amino acid substitution of an isoleucine for a threonine, and three high level resistant strains also showed a GAC to GGC mutation in codon 87, leading to the substitution of a glycine for an aspartic acid. In addition, four resistant strains also had an TCG to TTG mutation in codon 87 of parC gene, leading to the amino acid substitution of a serine for a leucine. The strains with both gyrA and parC mutations were two to sixteen times more resistant than the strains which had only gyrA mutations. At the same time, a silent mutation (CAC to CAT) in codon 132 of gyrA gene and a silent mutation(GCT to GCG) in codon 115 of parC gene occured, which did not lead to amino acid change. Conclusion The mutations of 83 and 87 codons of gyrA and the mutatations of 87 codon of parC gene were related to fluroquinolone resistance, and the mutations of the 83 codon of gyrA gene were more important.

The effect of chitin on human umbilical vein endothelial cells (HUVECs) injured by oxygen free radicals was investigated in vitro. The degree of injury was determined by the amount of malondialdehyde (MDA) and lactic dehydrogenase (LDH) in the culture medium. The results indicated that chitin could reduce the amount of MDA and LDH significantly, showing that chitin could protect HUVECs from oxygen free radicals injury.

In this paper, a qualitative and quantitative analysis method was established for the scopoletin in herba Artemisiae hedinii hy HPLC. The chromatographic column was Agilent TC-C18 (4.6 mm×250 mm, 5μm);the tem-perature of column was 30℃, eluted with a mobile phase consisted of methanol and 0.3%phosphoric acid solution and gradient elution at a flow rate of 1.0 mL·min-1. The detection wavelength was 344 nm. The results showed that the mass concentration and peak area of scopoletin had a good linear relationship within the range of measurement (r=0.999 9). The reproducibility was good;and the RSD was 1.51%. The average sample recovery rate was 101.42%. It was concluded that the established method was stable, reliable and simple. It provided theoretical evidences for the quality evaluation and quality control of herba A. hedinii.

OBJECTIVE To ascertain the causes of unreported hospital infection in our hospital and apply countermeasures. METHODS To adopt investigation way to review and analyze 34 694 hospital files in our hospital from Aug 2002 to Jul 2004. RESULTS From them 1164 cases had experienced hospital infection,264 were unreported,the monthly unreported rate decreased from 52.50% to 0;the yearly unreported rate decreased from 36.17% to 10.67%. CONCLUSIONS The unreported reason is because the relevant staff lack infection knowledge,the control and supporting system isn′t so efficient.So the key to reduce the missing report rate is to enhance the awareness of the hospital infection control among the staff in hospital,to strengthen communication and cooperation,and to implement the administrative regulation.

Objective To improve the awareness,diagnosis and treatment of pneumocystis carinii pneumonia (PCP) after renal transplantation.Methods A retrospective review was performed in 28 patients who underwent renal transplantation and developed PCP afterwards.The main clinical manifestations were fever(28 cases),nonproductive cough(28 cases),chest distress (12 cases).Occurrences of PCP were described 1.5 to 7 months after the renal transplantation.There were 10 patients treated with tacrolimus (FK506 2-6 rag/d,FK506 concentration 4-10 ng/ml) and 18 patients treated with cyclosporine (CsA 200-500 mg/d,CsA trough level:150-250 ng/ml) based immunosuppressive regimen.Anti-CD_(25)~+ monoclonal antibody (anti-CDCD_(25)~+mAb) was used in 10 cases for immune induction before operation while single steroid in 18 cases.Creatinine of patients with PCP was 70 to 106 μmol/L.CD_4~+ lymphocyte counts of the peripheral blood were 245±32/μl before PCP treatment and 536±25/μl after recovery.The most abnormal chest radiological findings were bilateral patchy ground-glass opacity.All the patients were diagnosed with PCP by bronchoalveolar lavage.Treatment was performed by reducing immunosuppressive agents and giving SMZco.Nineteen patients who had a PaP2 less than 70 mm Hg were given intravenous small-dose steroid.Results All the patients recovered from PCP 2 to 3 weeks after treatment.One patient experienced recurrence half year later.Five patients with higher creatinine after treatment recovered to normal levels after stopping the treatment of SMZco.No significant differences were seen in PCP patients treated with CsA and FK506,P＞0.05.The similar results were observed in use of anti-CDCD_(25)~+ mAb and single steroid,P＞0.05.Significant differences were observed in PCP patient peripheral blood CD_4~+ lymphocyte counts before and after treatment (P=0.001).Conclusions Patients who have fever,cough and hypoxia,chest imaging showing bilateral lung interstitial inflammation,might be PCP patients in the early post-renal transplantation period.Effective treatment should be performed by reducing immunosuppressive agents and giving SMZco.

Objective To evaluate the safety,feasibility and results of the hand-assisted retroperitoneal laparoscopic living donor nephrectomy ( HRPLDN ) with a modified technique. Methods Living donors (n =32) were divided into HRPLDN group (n =16) and open group (n =16) according to surgical technique.Operative data and postoperative outcomes including operative time,estimated blood loss,warm ischemia time,length of hospital stay and complication rate,were collected. Results All procedures were completed successfully.In HRPLDN group,the mean operative time was 101.3 ± 21.2 min (range from 70 to 150 min),with an estimated blood loss of 53.8 ±25.5 ml (range from 20 to 100 ml) and warm ischemia time of 2.4 ± 0.6 min ( range from 1.5 to 3.5 min).No living donor needed conversion to open surgery and the urine volume of transplanted kidney after first 24 hours was 5036 ml (range from 3500 -6500 ml).The mean postoperative on bed time were (2.8 ± 0.7 ) d (ranging from 2 -4 d).All parameters of HRPLDN were significantly better than that of open groups. Conclusion Living donor nephrectomy with HRPLDN is a safe and reliable surgical technique.

Objective To investigate the merit and demerit for surgical excision of polycystic kidney at the same time of kidney transplantation and to analyze its effect on complications and prognosis.Methods The data of 63 cases of polycystic kidney were retrospectively analyzed.Among the 63 recipients,43 recipients were combined with polycystic liver,and 2 with pancreatic cyst.For the large size of polycystic kidney,in 31 patients with hematuria or urinary tract infection,the polycystic kidney was resected during kidney transplantation(kidney-cut group,31 cases).The polycystic kidneys in The remaining 32 cases were preserved during kidney transplantation (reservation group).All the recipients were treated with CsA(Tacrolimus),mycophenolate mofetil (MMF)and prednisone after transplantation.The general conditions of recipients,the occurrence of delayed graft function(DGF),acute graft rejection.operative complications and infection,and survival rate of recipients and grafts were observed.Results Operative time in kidney-cut group was (300±31)min,and perirenal drainage tube duration was(4.6±1.4)days in kidney-cut group,significantly longer than in reservation group(both P＜0.01).Volume of red blood cells transfusion in kidney-cut group was(4.31±1.05)U,significantly more than reservation group(P＜0.01).29.0%(9/31)recipients in kidney-cut group had surgical complications,significantly higher than reservation group(6.2 0A,2/32)(P＜0.05).The ineidence of urinary tract irdection was 31.2% (10/32)in reservation group,significantly higher than in kidney-cut group(6.5%,2/31,P＜0.05).12.5%(4/32)patients in reservation group needed surgical excision of polycystic kidney after kidney transplantation due to polycystic kidney infection.In 24 recipients with preoperative high blood pressure in each group,the blood pressure of 8 recipients(33.3%)in kidney-cut group returned to normal,compared with only 2 recipients(8.3%)in reservation group(P＜0.05).The incidence of DGF,incidence of acute rejection,human/kidney 1-and 5-year survival rate between 2 groups had no statistically significant difference.Conclusion As long as the detailed operation.it is safe to resect patient's polycystic kidney during renal transplantation,but there are no effects on patient/kidney survival rate.

Objective:To detect the expression of microRNA-218 (miR-218) in human acute lymphocyte leukemia (ALL) T lymphocytes ( CCRF-CEM) ,explore its effects on the biological features of CCRF-CEM cells and the expression of its target gene c-kit, so as to provide new insights for leukemia treatment.Methods: Using the quantitative real-time polymerase chain reaction ( qRT-PCR) ,we detected the expression of miR-218 in the normal peripheral blood T lymphocytes and CCRF-CEM cells.Forty-eight hours after the miR-218 mimic was transfected into the CCRF-CEM cells,the expression of miR-218 in the CCRF-CEM cells was detected by qRT-PCR.The effect of miR-218 on the CCRF-CEM cell viability was detected using MTT.The effect of miR-218 on the proliferation and apoptosis of CCRF-CEM cell was analyzed using flow cytometry.c-kit gene was identified to be a target gene of miR-218 by luciferase reporter enzyme system,and the effect of miR-218 on the expression of KIT protein in cells were determined using Western blot.Results:As shown by qRT-PCR,compared with that in the normal peripheral blood T lymphocytes,the expressions of miR-218 in ALL T lymphocytes cell lines were significantly decreased ( P<0.01 ) .Compared with the control group, the expression of miR-218 increase significantly in CCRF-CEM cells transfected with miR-218 mimic for 48 hours ( P<0.01).MTT showed that the cell viability decreased significantly after the over-expression of miR-218 in the CCRF-CEM cells ( P<0.05 ) .Flow cytometry showed that the S-phase fraction significantly declined after the over-expression of miR-218 ( P<0.01 ) , and meanwhile the apoptosis of cells also significantly increased (P<0.01).Luciferase reporter gene assay showed that,compared with the control group,the relative luciferase activity significantly declined in the miR-218 mimic transfection group (P<0.01).Compared with the control group,the expression of KIT protein in the CCRF-CEM cells transfected with miR-218 mimic for 48 hours significantly decreased ( P<0.01).Conclusion:The expression of miR-218 decreases in ALL T lymphocytes cell lines.MiR-218 can negatively regulate the expression of KIT protein,inhibit the proliferation and increase the apoptosis of CCRF-CEM cells.Treatment based on the enhanced expression of miR-218 may be a promising strategy for leukemia.

Objective:To investigate the expression and significance of B cell activating factor (BAFF) and a proliferation-inducing ligand ( APRIL) in children with acute lymphoblastic leukemia ( ALL).Methods:The mRNA and protein expressions in ALL.

Objective To investigate the effect of Ser473-Akt phosphorylation in the protection of atorvastatin to cerebral ischemia-re-perfusion (I/R) injury in rats. Methods Forty male Sprague-Dawley rats were randomly divided into normal group (n=10), sham group (n=10), I/R group (n=10) and intervention group (n=10). A model of cerebral ischemia-reperfusion in rats was establishied, with ischemia for 2 hours and reperfusion for 72 hours. The normal group and the sham group received no injury. I/R group was administered with normal saline only, and the intervention group received atorvastatin 10 mg/kg prepared with normal saline at palinesthesia, 24 and 48 hours after reperfu-sion. All rats were sacrificed 72 hours after reperfusion. HE staining and TUNEL staining were performed in the brain specimens. The ex-pression of Akt and Ser473-Akt in the prefrontal cortex of the brain were detected with Western blotting. Results Compared with I/R group, 72 hours after reperfusion, the damage of nerve cells significantly lessened in the intervention group;the apoptosis positive cells significant-ly reduced in the intervention group (t=-6.014, P<0.001). The expression of Ser473-Akt in prefrontal cortex was higher in I/R group than in the normal group and the sham group (t>20.327, P<0.001), and was higher in the intervention group than in I/R group (t=3.649, P=0.007). Conclusion The Ser473-Akt phosphorylation plays an important role in the protection of atorvastatin in nerve cell through anti-apoptosis of nerve cells, and reducing cerebral I/R injury.