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.

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.

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 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 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 observe hypoxia-induced pulmonary arterial endothelial-to-mesenchymal transition and investigate the role of transforming growth factor β1 (TGF-β1) in the process. Methods Pulmonary arterial cells improved by adherence method were cultured in normoxia (containing 21%O2,5%CO2 and 74%N2) or hypoxia (containing 1%O2,5%CO2 and 94%N2) for 1,4,or 7 days,respectively. Endothelial-to-mesenchymal transition was confirmed with morphological observation and expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. Expression of TGF-β1 was evaluated by RT-PCR and Western blot,and α-SMA by Western blot. Results Hypoxia-induced paving-stone-like pulmonary arterial endothelial cells transdifferentiating to polygonal cells with high-expression of α-SMA. TGF-β1 expression was increased significantly after 7 days of hypoxia. TGF-β1 stimulating alone increasedα-SMA expression of pulmonary arterial en-dothelial cells;while,SD-208,inhibitor of TGF-β1,abolished the above effect. Conclusion Hypoxia can induce endothelial-to-mesenchymal transition. And TGF-β1 plays an important role in the process.

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.

Background Recent studies indicated that human amnion mesenchymal stem cells (hAMSCs) can be induced to differentiate into multiple types of cells in vitro,but whether the hAMSCs can differentiate into ocular surface cells has not been reported yet.Objective This study was to investigate the feasibility of inducing differentiation of hAMSCs into ocular surface cells by co-culturing with human bulbar conjunctiva fibroblasts (hBCFs).Methods This study was approved by Ethic Committee of Affiliated Second Hospital of Guangzhou Medical University.HAMSCs were isolated from placenta under the informed consent of healthy delivery women.hAMSCs were cultured,passaged and identified by detecting the expressions of CD44,CD45,CD73,CD90 in the cells with flow cytometer,osteogenesis and adipogenic differentiation experiments.Human conjunctival tissue was obtained during the eye operation under the informed consent of patients and hBCFs were isolated and cultured with explant culture.The cells were divided into the hAMSCs culture group and the hAMSCs and hBCFs co-culture group and cultivated in Transwell chambers for 7 days.The expressions of cytokeratin 19 (CK19) and α-smooth muscle actin (α-SMA) in the cells were assayed by immnofluorescence technique.Results Cultured hAMSCs showed the slender shape and cell body enlarged with passage.CD44,CD73 and CD90 were expressed in the cells,and the expression of CD45 was absent.After 3-4 weeks of osteogenesis and adipogenic induce,the cells showed red staining for alizarin and oil red O.In the co-culture group of hAMSCs and hBCFs,hAMSCs presented the epithelioid cell-like in shape and showed the positive response for CK19 and weaker response for α-SMA.However,in the hAMSCs culture group,the cells showed the positive response for α-SMA and absent response for CK19.Conclusions The hAMSCs can differentiate into ocular surface cells after being induced by hBCFs.And the differentiation mechanism is possibly relevant to mesenchymal cells epithelium.

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.