OBJECTIVE To investigate the repressor SmeT and other sequences of efflux pump SmeDEF in the clinic isolates of Stenotrophomonas maltophilia to involve in antibiotic resistance. METHODS The mRNA level of smeD examined the resistance profile of the clinical strains to antimicrobials was by agar dilution and their reaction to pump inhibitor was investigated by RT-PCR,and the smeD-smeT fragment was amplified by PCR,and then sequenced.Adding poly(A) to 3′ end of RNA,applying 3′RACE,nested PCR,then sequence-analysis. RESULTS The amino acid sequences of the forepart of SmeT were conservative in all the 7 strains.In antibiotic-resistant strains with positive reaction to pump inhibitor,the intergenic sequences of smeD-smeT were different from those in sensitive ones.The variations in(-165 to-82)nt of smeT and several nucleic acids before the start codon of smeT,possibly involved in antimicrobial resistance.All isolates didn′t have Leu166Gln mutation within the SmeT protein of D457R,which was reported possibly associated with antibiotic resistance.The resistant strains with efflux positively inhibited had Asp218Glu substitution,different from the negatively-inhibited ones. CONCLUSIONS The expression level of SmeDEF is related with the antibiotic resistance of S.maltophilia.The possible resistance mechanism includes the variations in the intergenic smeD-smeT and/or smeT codon sequence.

Objective To investigate the biological characteristics of recombinant hIFN-α-2B-BCG and its direct effect to bladder tumor cells in vitro. Methods BCG and recombinant BCG(rBCG) wild-type growth and morphology were compared. After 10 generations, hIFN-α-2B content was analyzed by ELISA, of bacteria in vivo and in vitro. The effects of rBCG on bladder cancer cells EJ, MB49, were detected by elec-tron microscopy and cell inhibitory rate from MTT. Results The normal BCG and rBCG had no significant difference in growth phase. They were both positive for acid-fast stain, maintaining the characteristics of cell's connection. While rBCG slightly larger than normal BCG, no else abnormality was obvious between them. IFN-α-2B was 997.2 pg/ml in secretions, 99.3 pg. In bacteria in vivo, 990.3 pg/ml in 10th genera-tion's sections, of rBCG. Compared with 1st generation, rBCG had no significant differences in morphology and interferon expression. Normal BCG and rBCG both had, anti-proliferation directly on bladder tumor cell in vitro, the rBCG is most effective in all. After rBCG cultivated with bladder tumor cell together, tumor cell's slow proliferation, detach, quantity decreasing and death were observed under microscope. Different degeneration in degree on most of tumor cells, disorganization on organelle, aggregation on chromatin, pyc-nosis on nucleolus, and cytoplasm lysis were on tumor cell under transmission electro microscopy. MTT as-say showed rBCG inhibited the proliferation of bladder caner cell and more active than normal BCG. Conclu-sion These results suggest that the rBCG have the same characteristics in growth phase as normal BCG, and stable properties in interferon expression and morphology by generations, rBCG has more anti-tumor effects in vitro than normal BCG.

Objective According to the characteristics of skin defects in ankle and foot select the posterior lateral leg pedide skin flaps supplyed by different blood to repair,providing advice and reference.Methods Between January,2001 and December,2015,163 cases of soft tissue defects at the foot and ankle were treated in our department.①The sural neurovascular flap was used in 93 cases in ankle and foot defect.②The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery in 36 cases in ankle and foot defect.③The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery including sensory reconstruction in 16 cases in heel rejion defect.④Peroneal artery perforator flap in 10 cases in anterior ankle,lateral malleolus and posterior malleolus defect.⑤The use of the sural fasciocutaneous flap alonged with peroneal artery and perforators in 8 cases in dorsal foot defect.The donor site with skin graft.Results Of the 93 sural neurovascular flaps,8 had partial loss,which were cured after dressing.All the sural nerve nutritional vessel flaps pediele with the perforating branch of the peroneal artery survived.Sensory grading standard by UK Medical Research Council was used to evaluate the recovery of sensory function on the last follow-up.The sensory function recovery of heel region flaps with reconstruction of the sensory was between S0 and S1.All peroneal artery perforator flaps survived.One of the sural fasciocutaneous flap alonged with peroneal artery and perforators occurred distal epidermis,which were cured after dressing.All patients were followed up 6-50 months (mean 20 months).All patients had recoveryed walking function,and infection wound had no recurrence after surgery.Conclusion According to the location,size,severity and the injury of peripheral vascular,select the most simple,safe,minimal damage flap for the soft tissue defects at the foot and ankle.The right choice and the exact design can improve flap survival rate,and recieve good clinical results.

BACKGROUND:Effective sorting of prostate cancer stem cels is the basis of experimental studies in prostate cancer developing. The most common sorting method is magnetic-activated cel sorting. OBJECTIVE:To separate CD133+/CD44+ cels in prostate cancer tissues using self-designed magnetic beads folowed by culture, passage and immunological identification. METHODS:Self-designed magnetic microspheres were applied to establish immunomagnetic beads to sort CD133+/CD44+ cels in prostate cancer tissues. The sorted cels were cultured in serum-free medium. The sphere formation, cel morphology, and proliferation ability after cel passage were statisticaly compared between the sorted cels and the normal tumor cel lines. Immunofluorescence detection was performed to detect the expression of specific antibodies. RESULTS AND CONCLUSION:Self-designed immunomagnetic beads had smal diameter and a high-sorting effect. The sorted cels possessed a high capacity of microsphere formation. After cel culture and passage, the cels highly expressed CD133 and CD44 antigens. The sorted cels with no induction had varying shapes and grew vigorously. After induction with transforming growth factor-β, the cultured cels were noted to have a single shape and grow slowly. The cel proliferation ability of sorted cels in these two groups differed significantly from that of the normal cancer cel lines (bothP < 0.05). In conclusion, the CD133+/CD44+ cels sorted from prostate tumor cels possessed cel morphology and function characteristics of stem cels which can provide a basis for extraction and culture of prostate cancer stem cels. Cite this article:Gong R, Li SY, Huo ZX, Ding H, Sun EL. Extraction of prostate cancer stem cels using self-designed magnetic beads. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):36-41.

Objective To investigate the effects of levofloxacin,ciprofloxacin,ceftazidime,pip-eracillin,cefoperazone/sulbactam,erythromycin,sulfamethoxazole and gentamycin on the bacterial biofilms of Stenotrophomonas mahophilia.Methotis Biofilm and conventional susceptibilities were determined for S.maltophilia isolates from 42 patients.The model of S.maltophilia biofflms in vitro was developed in the Mueller-Hinton broth--micmtiter inoculator or silica films.After antibiotic challenge plate 20 h,each plate was sonicated and the absorbance value at 620 nm(A620)was measured on a microtiter plate colorimeter be-fore and after incubation for 6 h.Then the biofilm inhibitory concentrations were calculated.Finally,based on the acquired data.the experiments of combinafion effects of erythromycin with the 3 antibiotic agents on the formed biofilms of 5 picked strains were designed and worked out.Results The sensitive rate of 42 S.maltaphilia to levofloxacin.sulfamethoxazole and piperacillin were 83.33%,66.67%and 54.76%,re-spectively.The bilfilm inhibitory concentrations were much higher than the corresponding minimal inhibitory concentrateion after formed biofflms.Conclusion Forty-two S.maltophilia are multi-resistant to antibiotic agent.And levofioxacin may have a better effect against biofilms compared with others.The inhibition effect of combination erythromycin with levofloxacin is more obvious among all the 3 antibiotic agents.

OBJECTIVETo explore the clinical diagnostic features and treatment of desmoplastic small round cell tumor (DSRCT), and to improve the understanding and management of this tumor.

METHODSThe clinicopathological data of nine patients treated in our hospital from October 2004 to June 2014 were retrospectively analyzed and a review of the literature was made. The clinical manifestations, pathological characteristics, diagnosis and differential diagnosis, treatment and prognosis of this tumor were summarized and analyzed.

RESULTSNine patients with DSRCT, 5 males and 4 females, with an average age of 21 years (range 8-56 years) were included in this study. Ultrasound examination revealed irregular low-density mass shadow in the abdominal cavity. CT examination found that 6 cases had abdominal and retroperitoneal multiple solid tumor nodules, uneven density, and visible low density fluid area. Postoperative pathological examination revealed that the tumor cells were small, mostly elliptic, gathered to form clear structure of nests with clear irregular boundaries. The central portion of large tumor nests often showed necrosis. Scattered fibroblasts and large amount of hyalinization of collagen fibers were seen in the interstitial tissue around the nests. Six patients received laparotomy surgery, however, all failed to resect the tumor completely. Three patients received postoperative chemotherapy, i. e. two cases had carboplatin and paclitaxel chemotherapy, and one case of chemotherapy regimen not specified. Two patients had radiation and chemotherapy (no concrete plan was available). Another case was lost to follow-up. Two of the three patients without surgery received chemotherapy with CAP (cyclophosphamide+adriamycin+carboplatin) and total rectal lesions, pelvic and inguinal lymph nodes, ilium metastases radiation therapy. Another one patient received EP regimen (DDP+VP16) which was then changed into a TP chemotherapy alone. Eight of the nine cases died shortly after surgery, and only one patient treated with chemotherapy alone was still alive after 11 months of follow-up.

CONCLUSIONSDesmoplastic small round cell tumor is a very rare, special type of soft tissue tumor, with very poor prognosis. This tumor may be preliminarily diagnosed according to the imaging characteristics and detection of tumor markers, however, final diagnosis is made by pathology. Surgery is the priority of treatment, combined with complementary radiation and chemotherapy.

Abdominal Neoplasms ; complications ; diagnosis ; mortality ; therapy ; Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; analysis ; Carboplatin ; administration & dosage ; Child ; Combined Modality Therapy ; methods ; Cyclophosphamide ; administration & dosage ; Desmoplastic Small Round Cell Tumor ; complications ; diagnosis ; mortality ; therapy ; Doxorubicin ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Paclitaxel ; administration & dosage ; analysis ; Prognosis ; Retrospective Studies