[Effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells and ciprofloxacin on Staphylococcus aureus in vitro].

Objective: To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on Staphylococcus aureus (SA) in vitro. Methods: hUCMSCs were isolated from umbilical cord tissue of full-term healthy fetus after cesarean section and cultured. Cells in the third passage were used in the experiments after identification. SA strains isolated from wounds of burn patients in our burn wards were used in the experiments. Cells were divided into 0, 10, 100, and 1 000 ng/mL lipopolysaccharide (LPS) groups according to the random number table (the same dividing method below). Cells were cultured with culture medium of mesenchymal stem cells (MSCs) after being treated with medium containing the corresponding mass concentrations of LPS for 12 h. At post culture hour (PCH) 6, 12, and 24, 6 wells of culture supernatant of cells in each group were obtained to measure the content of LL-37 with enzyme-linked immunosorbent assay. Ninety blood agar plates were divided into ciprofloxacin control group (CC), ciprofloxacin+ supernatant group (CS), and ciprofloxacin+ supernatant+ LL-37 antibody group (CSL), with 30 blood agar plates in each group. Blood agar plates in group CC were coated with 1.5×10(8) colony forming unit (CFU)/mL bacteria solution prepared with normal saline. Blood agar plates in group CS were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline and culture supernatant of hUCMSCs (cultured by culture medium of MSCs, the same below) in double volume of normal saline. Blood agar plates in group CSL were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline, culture supernatant of hUCMSCs in double volume of normal saline, and 2.6 µL LL-37 antibody in the concentration of 2 µg/mL. At PCH 12, 24, and 48, 10 blood agar plates of each group were harvested to observe the distribution of SA colony on blood agar plate and to measure the diameter of bacterial inhibition ring of ciprofloxacin. The minimum inhibitory concentration (MIC) of ciprofloxacin against SA of each group was recorded. Fractional inhibitory concentration (FIC) indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were calculated, and the effect of synergy was evaluated. Data were processed with analysis of variance of factorial design, one-way analysis of variance, LSD-t test, Kruskal-Wallis test, and Mann-Whitney U test. Results: (1) At each PCH, the content of LL-37 in culture supernatant of cells in 10, 100, and 1 000 ng/mL LPS groups was higher than that in 0 ng/mL LPS group (with t values from 11.22 to 33.36, P values below 0.01); the content of LL-37 in culture supernatant of cells in 100 and 1 000 ng/mL LPS groups was higher than that in 10 ng/mL LPS group (with t values from 2.24 to 18.73, P<0.05 or P<0.01); the content of LL-37 in culture supernatant of cells in 1 000 ng/mL LPS group was higher than that in 100 ng/mL LPS group (with t values from 12.46 to 14.70, P values below 0.01). (2) At PCH 12, 24, and 48, the bacterial colonies in groups CC, CS, and CSL began to integrate over time. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in group CC were 26, 24, and 23 mm, respectively, with no obvious change. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in groups CS and CSL were 82, 71, 68 mm, and 74, 59, 56 mm, respectively, significantly longer than those of group CC. (3) At each PCH, the MIC of ciprofloxacin against SA was significantly higher in group CC than in groups CS and CSL (with Z values from 6.22 to 6.71, P values below 0.01); the MIC of ciprofloxacin against SA was significantly higher in group CSL than in group CS (with Z values all equal to 6.72, P values below 0.01). (4) FIC indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were 0.011, 0.032, 0.032, and 0.122, 0.350, 0.350, respectively. The results indicated that culture supernatant of hUCMSCs had synergistically antibacterial effect on ciprofloxacin. Conclusions: hUCMSCs can secrete LL-37, and the secretion level is increased with increase of LPS concentration. Combination of culture supernatant of hUCMSCs and ciprofloxacin can decrease the dosage of ciprofloxacin in resisting SA. Once LL-37 is neutralized, the synergistically antibacterial effect of culture supernatant of hUCMSCs is decreased.

Predicting precipitation on nonpoint source pollutant exports in the source area of the Liao River, China.

The source area of the Liao River is an important grain growing area in China which experiences serious problems with agricultural nonpoint source pollution (NPS) which is impacting the regional economy and society. In order to address the water quality issues it is necessary to understand the spatial distribution of NPS in the Liao River source area. This issue has been investigated by coupling a wavelet artificial neural network (WA-ANN) precipitation model with a soil and water assessment tool (SWAT) model to assess the export of nonpoint source pollutants from the Liao River source area. The calibration and validation of these models are outlined. The WA-ANN models and the SWAT model were run to generate the spatial distribution of nonpoint source nutrient (nitrogen and phosphorus) exports in the source area of the Liao River. It was found that the SWAT model identified the sub-catchments which not only receive high rainfall but are also densely populated with high agricultural production from dry fields and paddy fields, which are large users of pesticides and chemical fertilizer, as the primary source areas for nutrient exports. It is also concluded that the coupled WA-ANN models and the SWAT model provide a tool which will inform the identification of NPS issues and will facilitate the identification of management practices to improve the water environments in the source area of the Liao River.

Surgical management of the hypopharyngeal and cervical esophageal cancer.

Between December 1979 and December 1992, 15 patients with hypopharyngeal and cervical esophageal cancer were treated surgically. Total pharyngolaryngectomy and partial cervical esophagectomy were performed and the defects were reconstructed with cervical skin flaps in 2 cases. The free jejunal segments were used in 6 cases following total pharyngolaryngectomy and cervical esophagectomy, cervical esophagectomy (larynx preserved) was repaired with free jejunal graft in 1 case. The pharyngogastric anastomosis following total pharyngolaryngoesophagectomy were performed in 4 cases, one of them, used pectoralis major myocutaneous flap for resection of soft tissue and skin of the neck. The pectoralis major myocutaneous flap and forearm free flap in 1 case respectively were used to reconstruct the deficits of total laryngectomy and partial pharyngectomy, and partial pharyngolaryngectomy. In our series, 1, 3, 5 years survival rates were 73.3% (11/15), 50% (6/12) and 55.6% (5/9), respectively. The advantages and disadvantages of a variety of operative procedures and the cervical lymph nodes management will be discussed.