The needle-recording electrodes deviced by the author and the improved recording method were applied in this article. Firstly, the lumbar spinal evoked potentials (LPs) and cortial somatosensory evoked potentials (CPs)to the stimulation of tiable nerves and segmental (radcular) cutaneous nerves (superficial peroneal and sural nerves )were recorded in 50 normal volunteers. Then similiar thacings were recorded and observed in 48 patients with unilateral L_5 or S_1 root compression. We found that the LP to the segmental cutaneous nerve stimulation was abnormal in 85%, and its CP in 48%. The LP to the tiable nerve stimulation was abnormal in 33%,and its CP only in 8%.

OBJECTIVETo study the application of VSD in the treatment of severe necrotizing fasciitis in extremities of patients.

METHODSEight 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.

CONCLUSIONSVSD 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.

Debridement ; Drainage ; Extremities ; surgery ; Fasciitis, Necrotizing ; surgery ; Granulation Tissue ; Humans ; Negative-Pressure Wound Therapy ; Oxygen ; Pressure ; Skin ; Skin Transplantation ; Ulcer ; Vacuum

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.

To determine whether the pathological changes caused by injury to the spinal cord can be correlated with values obtained by the Magnetic Motor Evoked Potential (MEPs) technique, we studied spinal cords from 41 adult cats who were divided into 4 groups. The groups ranged from normal cats whose spinal cords were not compressed, to slightly, moderately and severely injured. MEPs were recorded before compression and in 30 minutes, 6 hours, 1 week, 2 week and 4 week after the compression unit was installed. Pathological changes with increased pressure were seen in blood vessels, nerve cells and fibers, Nissl substance and the central canal. A reversal of pathological changes was observed in slight or moderate injury during the 4 weeks of the experiment. Extensive injury, however, caused irreversible changes in the nerve cells with loss of motor function. The latency of MEPs at 30 minutes and 6 hours in the slightly injured group was 0.37 and 0.38 times greater than the baseline and returned to normal levels in 4 weeks. In the moderately injured group, the latency was increased 0.77 and 0.81 times and in the severely injured 1.32 and 1.36 times over the baseline. Recovery in the second group was partial and not at all in the severely injured. Thus, there appears to be good correlation between observed pathological changes, motor functions and MEPs.

Objective To find out an effective therapeutic method for and observe whether there is any synergistic action or not between fetal spinal cord transplantation (FST) and methylprednisolone (MP).Methods Fifty male adult SD rats were randomly divided into group A,B,C,D and E,10 in each group.Group A was treated with both large dosage of MP and FST,group B with MP only, grop C with FST only and group D without any treatment.Group E served as blank control.Fetal spinal cord was obtained from 14-day pregnant rats .Spinal cord Somatosensory evoked potential (SSEP) examination and behavior observation were performed in 24 hours and in 8 months after treatment By the way of reduced silver staining, the condition of nerve plerosis and regeneration could be observed.Results There were significant differences in the latent period and amplitude of N1 wave in SSEP between group A and group B,C and D (P＜0.05).No obvious behavior changes were found except partial sensory recovery in the left lower limbs in Group A.Histologically,more nerve fibers contacting with branches at injury area could be found in Group A than in Group B,C and D.Conclusion The combination of large dosage of MP and FST can produce synergistic effect in the recovery of the injured spinal cord.

In order to observe the damaged nerve successively, we used superficial magnetic stimulation motor evoked potential (MEP) in the pathological model of chronic compressed nerve of the cervical nerve roots of cats. We synthesized various change of the pathomorphology of the nerve damaged to different degrees, and discussed the relationship between the MEP and the pathomorphologic change of chronic compressed nerve roots. The results showed that the initial pathologic change of nerve with myelin was degeneration of myelin shealth. The MEP of the nerve also showed increased latency and dispersed wave forms. After that the axon of the demylinated nerve degenerated, splitted and had a peripheral Wallerian Degeneration. The MEP showed an increased latency along with decreased amplitude. The degree of the MEP's change accompanyed with pathologic change. So we believe that the magnetic compressed nerve. It has some reference value in figuring out the damage by analysing factors.

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.

OBJECTIVETo 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.

METHODShECs 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.

RESULTS(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).

CONCLUSIONSThe 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.

Adenoviridae ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; EGF Family of Proteins ; genetics ; metabolism ; Epidermis ; cytology ; Genetic Vectors ; Humans ; Keratins ; metabolism ; Male ; Transfection

Objective To investigate the risk factors for myocardial injury after Nano-Knife surgery in patients with pancreatic cancer, and to establish a nomogram model for risk prediction. Methods A retrospective analysis was performed for the clinical data of 92 patients with pancreatic cancer who underwent Nano-Knife surgery in The Fifth Affiliated Hospital of Zhengzhou University from September 2020 to November 2021, with serum cardiac troponin I > 0.03 ng/mL within 3 days after surgery as the diagnostic criteria for myocardial injury, the patients were divided into myocardial injury group with 51 patients and non-myocardial injury group with 41 patients. Related baseline data were collected for all patients, including age, sex, body mass index, American Society of Anesthesiologists classification, smoking history, alcohol abuse history, and preoperative comorbidities. The Mann-Whitney U test was used for comparison of continuous data between groups, and the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups. Univariate and multivariate logistic regression analyses were used to screen out the variables with statistical significance, and the factors screened out were used to establish a nomogram for predicting the risk of myocardial injury after Nano-Knife surgery in patients with pancreatic cancer. The receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to evaluate the discriminatory ability and clinical utility of the model. Results Compared with the non-myocardial injury group, the myocardial injury group had a significantly longer ablation time ( χ 2 =7.410, P =0.006), a significantly greater number of probes ( χ 2 =6.130, P =0.047), and a significantly higher proportion of patients with preoperative hypertension ( χ 2 =12.124, P < 0.001) or chronic kidney disease ( χ 2 =12.829, P < 0.001). The univariate logistic regression analysis showed that tumor diameter, ablation time, surgical procedure, number of probes, history of hypertension, and history of chronic kidney disease were associated with the development of myocardial injury (all P < 0.05), and the multivariate logistic regression analysis showed that tumor diameter (odds ratio [ OR ]= 3.94, 95% confidence interval [ CI ]: 1.09-14.18, P =0.036), ablation time ( OR =4.15, 95% CI : 1.30-13.27, P =0.016), surgical procedure ( OR =6.92, 95% CI : 1.92-25.07, P =0.003), and history of hypertension ( OR =4.07, 95%CI: 1.12-14.77, P =0.034) were independent risk factors for myocardial injury after Nano-Knife surgery in patients with pancreatic cancer. An AUC of 0.859 showed that the nomogram had good discriminatory ability and clinical utility. Conclusion There is a relatively high incidence rate of myocardial injury after Nano-Knife surgery in patients with pancreatic cancer. Preoperative hypertension, tumor diameter > 4 cm, and ablation time > 1 hour are independent risk factors for myocardial injury, and the surgical procedure of Nano-Knife surgery and bypass anastomosis can increase the risk of myocardial injury. The nomogram has a good effect in predicting the risk of myocardial injury.