ABSTRACT
Objective:To analyze the value of low dose enteral nutrition (EN) in treatment of septic shock combined with acute gastrointestinal injury Ⅲ (AGI Ⅲ).Methods:Clinical data of septic shock patients combined with AGI Ⅲ admitted at our hospital were analyzed.Patients were divided into two groups according to the nutrition therapy they received:treatment group (EN,n =41) and control group (no EN,n =46).The mortality and ICU hospital stays were collected.The intestinal barrier,inflammatory cytokines,and oxidative stress were evaluated before and after EN treatment.Results:For patients in the treatment group,the dosages of EN ranged from 200 to 410 kcal/d,with the median dose of 350 kcal/d.No significant differences were found on death rates between the two groups (24.4％vs 32.6％,P =0.398).Patients in the treatment group had shorter ICU hospital stays than those of the control group (11.8 ± 3.7 vs 16.2 ± 5.3,P ＜0.01).After one week EN treatment,patients in the treatment group had lower levels of CRP,IL-6,TNF-α,diamine oxidase,endotoxin and D-lactate than those of the control group (P ＜ 0.05).Conclusion:For septic shock patients combined with AGI Ⅲ,low dose EN can improve the intestinal barrier function and systemic inflammatory responses.
ABSTRACT
Objective To investigate the therapeutic effect of Guizhi-Fuling pill combined with danazol for endometriosis and its possible therapeutic mechanisms. Methods A total of 316 patients with endometriosis were recruited and randomly divided into a routine treatment group and a combined treatment group, 158 in each group. The conventional treatment group were treated by oral danazol tablets, and the combined treatment group by oral danazol and Guizhi-Fuling pill for 28 days. The serum levels of vascular endothelial growth factor (VEGF), VEGF receptor Flk-1, estrogen receptorα(ERα), 17β-estradiol (E2), IL-2, IL-6 and IL-8 were detected by the enzyme-linked immunosorbent assay. The expressions of VEGF and Flk-1 mRNAs in the ectopic endometriotic stromal cells were determined with the reverse transcriptase-polymerase chain reaction. Results The total effective rate in the combined treatment group was significantly higher than that in the routine treatment group (93.7% vs. 78.5%; χ2=13.968, P<0.01), and the recurrence rate was significantly lower (5.4%vs. 13.7%;χ2=19.984, P<0.01). After the treatment, the serum levels of VEGF (2.93 ± 0.07 pg/ml vs. 6.85 ± 0.12 pg/ml;t=354.679, P<0.01), Flk-1 (2.97 ± 0.12 pg/ml vs. 5.87 ± 0.17 pg/ml;t=175.179, P<0.01), ERα(4.93 ± 0.29 pg/ml vs. 6.85 ± 0.37 pg/ml;t=51.337, P<0.01), E2 (11.69±1.09 pg/ml vs. 14.10 ± 0.78 pg/ml;t=22.601, P<0.01), IL-2 (2.27 ± 0.12 pg/ml vs. 2.31 ± 0.16 pg/ml;t=2.514, P<0.05), IL-6 (0.39 ± 0.03 pg/ml vs. 0.59 ± 0.06 pg/ml;t=37.476, P<0.01) and IL-8 (0.47 ± 0.04 pg/ml vs. 0.81 ± 0.09 pg/ml;t=43.393, P<0.01) in the combined treatment group were significantly lower than those in the routine treatment group;the expressions of VEGF (0.69 ± 0.08 vs. 2.11 ± 0.09; t=148.229, P<0.01) and Flk-1 (4.67 ± 0.14 vs. 5.61 ± 0.22;t=45.311, P<0.01) mRNAs in the ectopic endometriotic stromal cells in the combined treatment group were significantly lower than those in the routine treatment group. Conclusions Guizhi-Fuling pill combined with danazol can downregulate the expressions of VEGF and Flk-1 in the ectopic endometriotic stromal cells, reduce the serum levels of VEGF, Flk-1, ERαand E2, alleviate inflammation, and improve symptoms.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the suitable transfection condition of human epidermal cells (hECs) with human epidermal growth factor (EGF) gene by adenovirus vector (Ad-hEGF) and its effects on the biological characteristics of hECs.</p><p><b>METHODS</b>hECs were isolated from deprecated human fresh prepuce tissue of circumcision by enzyme digestion method and then sub-cultured. hECs of the third passage were used in the following experiments. (1) Cells were divided into non-transfection group and 5, 20, 50, 100, 150, and 200 fold transfection groups according to the random number table (the same grouping method below), with 3 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in the latter six groups were transfected with Ad-hEGF gene in multiplicities of infection (MOI) of 5, 20, 50, 100, 150, and 200 respectively. The morphology of the cells was observed with inverted phase contrast microscope, and expression of green fluorescent protein of the cells was observed with inverted fluorescence microscope at transfection hour (TH) 24, 48, and 72. (2) Another three batches of cells were collected, grouped, and treated as above, respectively. Then the transfection rate of Ad-hEGF gene was detected by flow cytometer (n=3), the mass concentration of EGF in culture supernatant of cells was detected by enzyme-linked immunosorbent assay (n=6), and the proliferation activity of cells was detected by cell counting kit 8 (CCK8) and microplate reader (n=6) at TH 24, 48, and 72, respectively. (3) Cells were collected and divided into non-transfection group and transfection group, with 6 wells in each group. Cells in non-transfection group were cultured with culture supernatant of cells without transfection, while cells in transfection group were cultured with culture supernatant of cells which were transfected with Ad-hEGF gene in the optimum MOI (50). CCK8 and microplate reader were used to measure the biological activity of EGF secreted by cells on culture day 1, 3, and 5. (4) Cells were collected and divided into non-transfection group and transfection group, with 12 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in transfection group were transfected with Ad-hEGF gene in the optimum MOI (50). The expression levels of cytokeratin 14 (CK14) and CK19 of cells were measured by immunofluorescence staining at TH 24. (5) Cells were collected, grouped, and treated as in (4), with 6 wells in each group. At post scratch hour (PSH) 0 (immediately after scratch), 12, 24, and 48, the migration distance of cells was observed and measured with inverted phase contrast microscope. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, and LSD test.</p><p><b>RESULTS</b>(1) At TH 24 and 48, morphology of cells in each transfection group and non-transfection group were similar. Compared with that in non-transfection group, the cell debris increased significantly in 200 fold transfection group at TH 72. At TH 24, 48, and 72, the expression of green fluorescent protein was not seen in cells of non-transfection group, whereas it increased in cells of transfection group over transfection time. (2) The transfection rate of Ad-hEGF gene of cells in each transfection group increased gradually over transfection time. At TH 72, the transfection rates of Ad-hEGF gene of cells in 50-200 fold transfection groups were all above 90%, while the transfection rates of Ad-hEGF gene of cells in non-transfection group, 5, and 20 fold transfection groups were (0.51±0.20)%, (62.44±6.23)%, and (75.00±5.43)% respectively, which were obviously lower than the rate in 50 fold transfection group [(93.12±2.55)%, with P values below 0.01]. The mass concentration of EGF in culture supernatant of cells in each transfection group increased gradually over transfection time. At TH 72, the mass concentration of EGF in culture supernatant of cells in 50 fold transfection group was obviously higher than that in each of the other groups (with P values below 0.01). The proliferation activity of cells in each group at TH 24 and 48 was similar (with P values above 0.05). At TH 72, the proliferation activity of cells in 200 fold transfection group was obviously lower than that in other groups (with P values below 0.05). (3) On culture day 1, the biological activity of EGF secreted by cells in two groups was similar (P>0.05). On culture day 3 and 5, the biological activity of EGF secreted by cells in transfection group were obviously higher than that in non-transfection group (with P values below 0.01). (4) At TH 24, the expression levels of CK14 and CK19 of cells in transfection group were higher than those in non-transfection group. (5) The width of scratch in two groups was nearly the same at PSH 0. At PSH 12-48, the migration distance of cells in transfection group was obviously longer than that in non-transfection group (with P values below 0.01).</p><p><b>CONCLUSIONS</b>The suitable range of MOI of hECs transfected with Ad-hEGF gene is 50-150, and 50 is the optimum. hECs transfected with Ad-hEGF gene with MOI 50 can effectively express the EGF gene and keep its good abilities of proliferation, differentiation, and migration, as well.</p>
Subject(s)
Humans , Male , Adenoviridae , Cell Differentiation , Cell Proliferation , Cells, Cultured , EGF Family of Proteins , Genetics , Metabolism , Epidermis , Cell Biology , Genetic Vectors , Keratins , Metabolism , TransfectionABSTRACT
PurposeTo optimize the imaging parameters of clinical MRI scanner in rat pancreas imaging to improve the image quality and to provide better MRI image quality and more economical research method for imaging study of rat pancreas. Materials and Methods Twenty-four healthy male Wistar rats were randomly divided into the conventional sequence (CS) group, the adjustment sequence (AS) group and the optimization sequence (OS) group, with 8 rats in each group. The rats in the CS group were scanned with conventional parameters using a clinical MRI scanner. The principle of parameter adjustment was: parameters associated with T1WI or T2WI imaging quality (TR, TE, slice thickness, NEX, FOV and matrix) was set with four changes, and only one of the six parameters was changed in each scan, image quality was evaluated by two senior radiologists, the parameter corresponded the best image quality evaluated consistently by two radiologists were selected as the optimal imaging parameter, all the optimized parameters were set up step by step in this way which formed the imaging parameters in OS group. The pancreatic signal intensity and signal to noise ratio was compared between CS group and OS group after imaging.Results The optimized sequence parameters in clinical MRI scanner were listed below: T1WI sequence (M3D/FSPGR/15): TR 6 ms, TE 2.5 ms, slice thickness 2.0 mm, NEX 8, FOV 7 cm×7 cm, Matrix 120×120; T2WI sequence (FSE-XL/90): TR 4000 ms, TE 71 ms, slice thickness 2.0 mm, NEX 1, FOV 8 cm×8 cm, Matrix 192×160. The pancreatic SI in T1WI and T2WI sequence of the OS group were significantly higher than those in the CS group (t=5.16 and 3.80,P<0.01), while the pancreatic SNR in T1WI and T2WI sequence of the OS group were significantly higher than those in the CS group (t=5.65 and 3.26,P<0.01).Conclusion The optimized parameters can improve the imaging quality of rat pancreas MRI significantly, thus provide a reference for the related experimental study.
ABSTRACT
<p><b>OBJECTIVE</b>To study the application of VSD in the treatment of severe necrotizing fasciitis in extremities of patients.</p><p><b>METHODS</b>Eight patients, suffering from severe necrotizing fasciitis, who had been traditionally treated with iodophor-soaked gauze for 21 to 365 days in other hospitals, were transferred to our institute because of the nonhealing wounds and systemic toxic symptoms induced by infection, from January 2011 to August 2013. After admission, surgical debridement was performed timely, and the necrotic tissue was collected during the operation for pathological observation after HE staining. After the operation, VSD was started with negative pressure ranging from -100 to -80 kPa, and the furacilin solution (0.2 g/L) and oxygen (2 L/min) were continuously infused into the wound during the treatment. Surgical debridement was performed repeatedly according to the wound condition followed by change of VSD dressings to continue VSD treatment. The wounds were closed by suturing or with autologous skin grafts after being covered by fresh granulation tissue. The times of surgical debridement, times of change of VSD materials, wound healing status, and length of stay in our institute were recorded. All patients were followed up for a long time. Results HE staining showed that there were diffuse necrotic adipose and fibrous connective tissues in the necrotic tissue, and the normal tissue structure disappeared accompanied by significant infiltration of inflammatory cells. The number of surgical debridement was 2 to 10 (3.9 +/- 2.8) times. The number of VSD materials change was 2 to 10 (4.0 +/- 2.9) times. Wounds were closed by suturing and healed in two patients; wounds in the other six patients were partially sutured, their residual wounds were healed by autologous skin grafting. The length of stay in our institute was 20 to 49 (33 +/- 10) days. All patients were discharged after recovery. Patients were followed up for 2 to 24 months, and their wounds were found to be in good condition without ulceration or recurrence.</p><p><b>CONCLUSIONS</b>VSD can effectively remove the necrotic tissues and exudates from the fascial spaces and promote proliferation of granulation tissue. Therefore it serves as an effective approach to the treatment of severe necrotizing fasciitis in extremities.</p>