Objective To establish human lung adenocarcinoma multidrug resistance cell lines in vitro,observe their biological characteristics,and investigate the mRNA expressions of DNA pol?,mdr 1,mrp1,GST-?,lrp and topo Ⅱ genes.Methods Paclitaxel-resistant cell lines(A549/TXL20) were established in vitro by exposure to stepwise increased concentrations of the drug in a cell culture medium.Biological morphology and cell cycles were analyzed by morphometry and flow cytometry.The chemoresistance indexes of cells were measured by methyl tetrazolium assay.Evaluation of growth and in vitro drug sensitivity were performed.RT-PCR was employed to analyze the mRNA expressions of the DNA pol?,mdr 1,mrp1,GST-?,lrp,and topo Ⅱ genes.Results ① Compared with parent cells,the resistant sublines had a lower confluent density.They were smaller and mixed with giant cells in different sizes and with different numbers of nucleoli,and the growth property of A549/TXL20 did not change significantly compared with A549 cell lines.② The resistant cells,A549/TXL20,were 19.3 times more resistant to paclitaxel and 67.4 times more resistant to cisplatin than the parent cells,and also demonstrated cross-resistance to mitomycin,vinblastine,and 5-fluouracil(5-FU). ③ Compared with the A549 celllines,an unreasonably higher level of drug resistance and lower drug concentration was detected in A549/TXL20 cells after exposure to the drug in the culture medium.④ The mRNA expression level of DNA pol?,mdr1,GST-?,mrp1 andlrp genes in A549/TXL20 cells was significantly higher than that in A549 cell lines(P

Background The pathogenesis mechanism of diabetic cataract has not been fully elucidated.Researches showed that multiple biological pathways participate in the pathogenesis of diabetic cataract,including oxidative stress.Astaxanthin can inhibit oxidative stress-mediated injury and lipid peroxidation.However,whether astaxanthin has the preventive effects on diabetic cataract is unclear.Objective This study was to investigate the preventive effects of astaxanthin on metabolic cataract in type 1 diabetic rats.Methods Thirty-eight 6-week-old SPF male SD rats were used in this study,and 1％ streptozocin was intraperitoneally injected to establish type 1 diabetic models in 30 rats,and 24 successful models were assigned to diabetic model group,low-dose astaxanthin group and high-dose astaxanthin group.Equal volume of normal saline solution was injected in the same way in 8 rats as the normal control group.Mixture foods containing 50 mg/(kg · day) or 100 mg/(kg · day) astaxanthin with olive oil and fodder were used continuously for 3 months in the rats of low-dose astaxanthin group and high-dose astaxanthin group,respectively,and mixture food of olive oil with fodder was used in the diabetic model group.Only fodder was used in the same way in the rats of the normal control group.The opacification of lens was examined by slit lamp section radiography system and graded on a scale of 1-5.The specimen of lens were prepared for the hematoxylin & eosin stain.The expression and lation of advanced glycosylation end products (AGEs) in the lens was examined using immunochemistry.The contents of oxidative stress-related indicators in the lens,such as AGEs,malonydialdehyde (MDA),catalase (CAT),superoxide dismutase (SOD) and mass fraction of glutathione (GSH),were assayed by ELISA.The experimental process complied with the national standard (Laboratory Animal Requirements of Environment and Housing Facilities [GB14925-2001]).Results The blood glucose levels of the rats were significantly higher in the diabetic model group,low-dose astaxanthin group and high-dose astaxanthin group than those in the normal control group at 2,4,6,8,10 and 12 weeks after modeling (all at P＜0.05),while the blood glucose levels of rats were not evidently different between low-dose astaxanthin group and high-dose astaxanthin group at various time points(all at P＞0.05).The rat lenses were transparent in the normal control group with scale of grade 1,and serious lens opacification was seen in the rats of the diabatic model group,with the scale of grade 5,while the rat lenses in the low-dose astaxanthin group and high-dose astaxanthin group were in grade 3-4.The contents of AGEs in the lenses were (7.23 ±0.50) μg/ml and (7.01 ±0.37) μg/ml,and M DA contents were (1.43 ± 0.22) mmol/L and (1.35±0.16)mmol/L in the low-dose astaxanthin group and high-dose astaxanthin group respectively,which were significantly lower than (7.61± 0.45) μg/ml and (1.62 ±0.42) mmol/L in the normal control group (all at P＜0.05).GSH contents in rat lenes were (272.70±12.53) ng/L and (283.52±16.17) ng/L,and SOD coneents were (55.45± 6.47) μmol/(min · L) and (56.73±5.12) μmol/(min · L),and CAT concents were (2.91 ±0.41) μmol/(min · L)and (3.02±0.13)μmol/ (min · L) in the low-dose astaxanthin group and high-dose astaxanthin group respectively,which were significantly higher than (241.52 ± 15.13) ng/L,(51.67 ± 5.45) μmol/(min · L) and (2.72 ± 0.27)μmol/(min · L) in the normal control group (all at P＜0.05).The GSH concent and SOD concent in rat lens were lower in the low-dose astaxanthin group than that in the high-dose astaxanthin group (both at P＜0.05).Conclusions Astaxanthin can postpone the pathogenesis and development of diabetic cataract in type 1 diabetic rats by antioxydative stress.

OBJECTIVE: To detect the active efflux gene qac gene in methicillin-resistant Staphylococcus aureus (MRSA) by hem-nested polymerase chain reaction (PCR) and to learn the carrier condition of qac gene. METHODS: The active efflux gene qacA/B and qacB of 80 strains MRSA isolated from clinical specimens from Aug 2006 to March 2008 were amplified in vitro by hem-nested PCR with the primers designed by computers based on qac information of Genbank, and the PCR fragments were sequenced and analyzed. RESULTS: We detected qacA/B in 19 out of the 80 MRSA strains (23.75%) and qacB in 18 out of the 80 MRSA strains (22.5%). Compared with sequences of qacA (NO.X56628) and qacB(NO.AF535087) in the Genbank, 98% and 97% were identical, respectively. CONCLUSION The active efflux gene qac gene in MRSA is detected by hem-nested PCR. The proportion of qac gene positive strains is high in clinical practice, which is related to its multi-antibiotic resistance.
Bacterial Proteins ; analysis ; genetics ; Base Sequence ; Drug Resistance, Multiple ; genetics ; Membrane Transport Proteins ; analysis ; genetics ; Methicillin-Resistant Staphylococcus aureus ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods

Objective:To evaluate the clinical efficacy of the bridge pedicled transplantation of medial leg fascial flap combined with medial hemisoleus muscle flap for contralateral leg soft tissue defect.Methods:Between January of 2012 and January of 2018, 12 patients with soft tissue defect of the leg were treated with bridge pedicled transplantation of contralateral medial leg fascial flap combined with medial hemisoleus muscle flap by posterior tibial artery. There were 9 males and 3 female, aged from 19 to 53 years (mean, 35 years). The size of the soft-tissue defects ranged from 12 cm×8 cm to 18 cm×9 cm. The immediate coverage of the fascial and muscle flaps and vessel pedicle were repaired by a meshed split-thickness skin graft. The donor site was closed directly. After the transplantation of the one pedicle and two flaps survived, vascular pedicle was cut off.Results:All the fascial and muscle flaps survived completely. No clinical vascular deficiency was found on the fascial and muscle flaps postoperatively. One case developed distal muscle flap small skin graft necrosis, and spontaneous healed after 2 weeks of dressing change. Follow-up period ranged from 2.5 to 4.5 years (mean 3.8 years). A good contour was confirmed both at the recipient and donor sites. Satisfactory clinical results were obtained in this series.Conclusion:This method is suitable for the treatment of soft tissue defects of the leg with only one major blood vessel, which reduces the damage to the donor site.

Objective:To evaluate the efficacy of the repairing anterior tibial adjacent double wounds by transposition with single-pedicle two flaps of the medial head of gastrocnemius.Methods:Between January of 2012 and January of 2018, 10 patients with the anterior tibial adjacent double wounds (7 males and 3 female, aged from 21 to 45 years) were treated by transposition with single-pedicle two flaps of the medial head of gastrocnemius. The size of the soft-tissue defects ranged from 2.0 cm×2.5 cm to 4.5 cm×4.0 cm. The medial head of the gastrocnemius was divided into two flaps with a single pedicle to repair two adjacent wounds of the anterior tibial. The muscle flaps were immediately covered by a meshed split-thickness skin graft, and the wound in donor site was closed directly.Results:All the muscle flaps survived completely. No clinical vascular deficiency was found on the muscle flaps postoperatively. Small wound dehiscence was developed in one patient and spontaneously healed 2 weeks after dressing change. Patients were followed up for 2.0 to 4.5 years. A good contour was confirmed at the recipient and donor sites. Satisfactory clinical results were obtained in this series.Conclusion:This method is suitable for the repair of two adjacent small wounds of the anterior tibial which can reduce the damage to the donor site.