To investigate the effect of OGT expression inhibited by RNAi on the alteration of tau phosphorylation and glycosylation level in HEK293T cells.The siRNAs targeting OGTgene(OGT-siRNA1～3) were designed and chemically synthesized.The OGT-siRNA1～3 were transfected into HEK293T cells via lipofectamine2000.The efficacy of RNA interference was detected by RT-PCR.The fluorescence of GFP/OGT was counted by fluorescence microscopy after pEGFP/OGT and OGT-siRNA1～3 cotransfected to HEK293T cells for 24 h.The level of tau phosphorylation and glycosylation were detected by Western blot after Plasmid pCI/tau441 and the siRNA cotransfected HEK293T cells for 48 h.OGT-siRNA3(100 nmol/L) could effectively downregulate the expression of OGT gene compared with Mock group(80.0% at mRNA level and 51.3% at protein level).The level of phosphorylation at various sites of the tau was significantly upregulated and the glycosylation was downregulated following the inhibition of OGT gene expression.There is an apparent negative correlation between the modification of phosphorylation and glycosylation of tau protein, the deficient in glucose uptaken/metabolism may be an important pathogenesis of AD.

Objective: To explore the application value of computed tomography angiography (CTA) and three-dimensional reconstruction in repairing high-voltage electrical burn wounds in necks, shoulders, axillas, and upper arms with tissue flaps. Methods: From December 2014 to December 2018, 12 patients with high-voltage electrical burns in necks, shoulders, axillas, and upper arms were hospitalized. The size of wounds ranged from 13 cm×10 cm to 32 cm×15 cm after complete debridement. Before tissue flap repair, the subclavian artery-axillary artery-brachial artery and their branches were examined by CTA. The main target vessels and their branches were conducted by three-dimensional reconstruction, and the development of the axis vessels for the tissue flaps planning to dissect and their branches were observed. For wounds in upper arms, amputation stump bone exposed wounds, and wounds in axillas and the anterior, the latissimus dorsi myocutaneous flap is the first choice for repair, if the thoracodorsal artery and internal and external branches are well developed according to CTA examination. Latissimus dorsi myocutaneous flaps were used in 6 patients with the area of myocutaneous flap ranging from 16 cm×12 cm to 32 cm×17 cm. All the donor sites were covered by split-thickness skin graft of thighs. For large wounds in occiputs, necks, and scapulas, the contralateral lower trapezius myocutaneous flap is the first choice for repair, if the superficial descending branch and deep branch of the contralateral transverse cervical artery are well developed according to CTA examination. For small wounds in necks and scapulas, the ipsilateral lower trapezius myocutaneous flap can be used for repair, if the superficial descending branch of the ipsilateral transverse cervical artery is well developed according to CTA examination. Lower trapezius myocutaneous flaps were used in 4 patients with the area of myocutaneous flap ranging from 18 cm×12 cm to 25 cm×17 cm. The donor site of one patient was sutured directly and the donor site of the other 3 patients was covered by split-thickness skin graft of thighs. For wounds in the posteromedial side of upper arms and the anterior side of axillas, the lateral thoracic skin flaps can be used for repair, if the latissimus dorsi myocutaneous flap can not be utilized for reasons of back burn or no muscle is needed for dead space, when the blood supply of side chest skin is reliable according to CTA examination. Lateral thoracic skin flaps were used in 2 patients with the area of skin flap ranging from 16 cm×12 cm to 17 cm×14 cm. The donor site of one patient was sutured directly and the donor site of the other one patient was covered by split-thickness skin graft of thigh. Results: During the operation of tissue flap repair in 12 patients, the orientation and starting position of the axis vessels were consistent with those observed by CTA examination before operation. All the tissue flaps survived after operation. During follow-up of 1 to 24 months, the patients were satisfied with no serious scar contracture affecting the function nor secondary infection or chronic ulcer. Conclusions CTA and its three-dimensional reconstruction technique can clearly reconstruct the subclavian artery-axillary artery-brachial artery and their branches before repair of high-voltage burn wounds in necks, shoulders, axillas, and upper arms. It can be used to observe whether the vessels are embolized or not and the starting position and orientation of blood vessels, which can provide an important reference for the selection of tissue flap transplantion.

Objective: To observe the analgesic and sedative effect and safety of application of dexmedetomidine combined with remifentanil in dressing change of conscious patients with non-intubation in burn intensive care unit. Methods: Forty patients conforming to the study criteria hospitalized in our burn intensive care unit from April 2015 to April 2017 were selected. Prospective, randomized, and double-blind method was used for the design. Patients were divided into dexmedetomidine group and dexmedetomidine+ remifentanil group according to the random number table, with 20 cases in each group. Patients in the two groups were respectively given corresponding drugs during dressing change. The frequency and time of dressing change, Verbal Rating Scale (VRS) score of patients during dressing change (at drug administration for 25 minutes) and after dressing change (25 min after dressing change), Ramsay Sedation Score (RSS) during dressing change, satisfaction level for anesthesia of the patients and physicians after dressing change, dosage of remifentanil, and various adverse effects during and after dressing change were recorded. The heart rate, mean arterial blood pressure (MAP), respiratory rate, and pulse oxygen saturation (SpO₂) before drug administration and at 10, 15, and 25 minutes after drug administration were also recorded. Data were processed with analysis of variance for repeated measurement, t test, chi-square test, and Fisher′s exact probability test. Results: (1) Totally 38 patients completed the trial. There were no statistically significant differences between patients in two groups in gender, American Association of Anesthesiologist Grading, age, weight, and total burn area (χ²=0.230, 0.146, t=0.224, 0.351, 0.367, P>0.05). (2) The frequency of dressing change of patients in two groups were both 48 times. The time of dressing change and VRS scores during dressing change of patients in two groups were similar (t=0.821, 1.522, P>0.05). The VRS score of patients in dexmedetomidine+ remifentanil group after dressing change was (3.1±0.4) points, obviously lower than (3.8±0.8) points in remifentanil group (t=2.213, P<0.05). The RSS, satisfaction level scores for anesthesia of the patients and physicians after dressing change in dexmedetomidine+ remifentanil group were (3.13±0.32), (3.44±0.41), and (3.13±0.25) points, respectively, obviously better than (1.82±0.24), (2.71±0.23), (2.53±0.41) points in remifentanil group (t=2.226, 2.684, 7.702, P<0.01). The dosage of remifentanil of patients in dexmedetomidine+ remifentanil group was (282±19) μg, obviously less than (340±31) μg in remifentanil group (t=9.896, P<0.01). There were no statistically significant differences between patients in two groups in rates of respiratory inhibition and hypotension (χ²=0.211, 0.154, P>0.05). Compared with those in remifentanil group, the rates of nausea, vomiting, and other gastrointestinal symptoms of patients in dexmedetomidine+ remifentanil group were obviously reduced (P<0.05), but the rate of bradycardia was obviously increased (χ²=6.008, P<0.05). (3) There were no statistically significant differences between patients in two groups in heart rate, MAP, respiratory frequency, and SpO₂ before drug administration (t=0.444, 0.892, 1.059, 1.039, P>0.05). The heart rates of patients in dexmedetomidine+ remifentanil group at 10, 15, and 25 minutes after drug administration were (83±11), (78±10), and (82±14) times per minute, respectively, significantly lower than (95±10), (87±12), and (89±12) times per minute in remifentanil group (t=5.592, 3.992, 2.630, P<0.05 or P<0.01). The MAP of patients in dexmedetomidine+ remifentanil group at 15 and 25 minutes after drug administration were (69.4±3.1) and (73.8±2.2) mmHg (1 mmHg=0.133 kPa), respectively, significantly lower than (75.4±3.0) and (78.1±3.5) mmHg in remifentanil group (t=9.181, 7.206, P<0.01). There were no statistically significant differences between patients in two groups in respiratory frequency at each time point after drug administration (t=1.489, 1.862, 1.963, P>0.05). The SpO₂ of patients in dexmedetomidine+ remifentanil group at 15 minutes after drug administration was 0.972±0.018, obviously lower than 0.979±0.015 in remifentanil group (t=2.070, P<0.05). Conclusions Application of remifentanil with small dosage has effective analgesia for conscious burn patients with non-intubation during dressing changes, however, adverse effects such as nausea and vomiting are likely to occur. Remifentanil combined with dexmedetomidine not only guarantee the analgesic effect, but also reduce the dosage of analgesics, improve the sedative effect and satisfaction of the patients for anesthesia, and reduce various adverse effects. However, it will increase the incidence of bradycardia and has some inhibition effect on circulation at the same time.

Objective: To observe the effects of arnebia root oil on wound healing of rats with full-thickness skin defect, and to explore the related mechanism. Methods: Eighty SD rats were divided into arnebia root oil group and control group according to the random number table, with 40 rats in each group, then full-thickness skin wounds with area of 3 cm×3 cm were inflicted on the back of each rat. Wounds of rats in arnebia root oil group and control group were treated with sterile medical gauze and bandage package infiltrated with arnebia root oil gauze or Vaseline gauze, respectively, with dressing change of once every two days. On post injury day (PID) 3, 7, 14, and 21, 10 rats in each group were sacrificed respectively for general observation and calculation of wound healing rate. The tissue samples of unhealed wound were collected for observation of histomorphological change with HE staining, observation of expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) with immunohistochemical staining, and determination of mRNA expressions of VEGF and bFGF with real time fluorescent quantitive reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, t test, and Bonferroni correction. Results: (1) On PID 3, there were a few secretions in wounds of rats in the two groups. On PID 7, there were fewer secretions and more granulation tissue in wounds of rats in arnebia root oil group, while there were more secretions and less granulation tissue in wounds of rats in control group. On PID 14, most of the wounds of rats in arnebia root oil group were healed and there was much red granulation tissue in unhealed wounds, while part of wounds of rats in control group was healed and there were a few secretions and less granulation tissue in unhealed wounds. On PID 21, wounds of rats in arnebia root oil group were basically healed, while there were still some unhealed wounds of rats in control group. (2) On PID 3 and 7, the wound healing rates of rats in arnebia root oil group were (39±5)% and (46±4)% respectively, which were close to (34±3)% and (44±4)% of rats in control group (with t values respectively 0.807 and 0.481, P values above 0.05). On PID 14 and 21, the wound healing rates of rats in arnebia root oil group were (76±4)% and (90±3)% respectively, which were significantly higher than (60±6)% and (73±5)% of rats in control group (with t values respectively 2.308 and 3.072, P<0.05 or P<0.01). (3) On PID 3, 7, and 14, granulation tissue, fibroblasts, and nascent capillaries in unhealed wound tissue of rats in the two groups both gradually increased, and more ranulation tissue, fibroblasts, and nascent capillaries were seen in unhealed wound tissue of rats in arnebia root oil group. On PID 21, granulation tissue, fibroblasts, and nascent capillaries in unhealed wound tissue of rats in the two groups both gradually decreased. (4) On PID 3, 7, and 14, the numbers of VEGF positive cells and bFGF positive cells in unhealed wound tissue of rats in the two groups both gradually increased; there were more VEGF positive cells and bFGF positive cells in unhealed wound tissue of rats in arnebia root oil group than those in control group. On PID 21, positive expressions of VEGF and bFGF both decreased in unhealed wound tissue of rats in the two groups. (5) On PID 3, 7, and 14, mRNA expressions of VEGF in unhealed wound tissue of rats in arnebia root oil group were higher than those of control group (with t values from 2.967 to 4.173, P values below 0.01). On PID 21, mRNA expression of VEGF in unhealed wound tissue of rats in arnebia root oil group was lower than that of control group (t=-4.786, P<0.001). From PID 3 to 21, mRNA expressions of bFGF in unhealed wound tissue of rats in arnebia root oil group were higher than those of control group (with t values from 2.326 to 4.702, P<0.05 or P<0.01). Conclusions Arnebia root oil can promote wound healing of rats with full-thickness skin defect, which may relate to increasing expressions of VEGF and bFGF.

OBJECTIVETo compare the accuracy of intaoperative methylene blue alone and in combination with (99m)Tc-sulfur colloid isotopic tracing for detection of sentinel lymph nodes (SLNs) in early-stage non-small cell lung cancer (NSCLC).

METHODSSixty-one patients with operable NSCLC who did not receive previous radiotherapy or chemotherapy were enrolled. Methylene blue and (99m)Tc-sulfur colloid were injected into the subserosal layer adjacent to the tumor, and SLNs were defined as those with blue staining or those containing 3 times more radioactivity than the surrounding tissue detected with a gamma probe. The SLN were removed with systematic lymph node dissection. All the removed lymph nodes were examined histopathologically with HE staining and immunohistochemistry.

RESULTSMethylene blue alone showed a low detection rate (60.0%) and sensitivity (58.33%) for SLNs compared with the combination of methylene blue and isotope tracing (96.15% and 92.86%, respectively).

CONCLUSIONThe combination of methylene blue and (99m)Tc-sulfur colloid isotopic tracing allows accurate detection of the SLNs in early-stage NSCLC.

Carcinoma, Non-Small-Cell Lung ; diagnosis ; Colloids ; Humans ; Immunohistochemistry ; Isotopes ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Methylene Blue ; Sentinel Lymph Node Biopsy ; Sulfur ; Technetium Tc 99m Sulfur Colloid

BACKGROUND:Abnormal extracellular matrix accumulation and excessive proliferation of fibroblasts are the main manifestations of pathological scars.Excessive proliferation of fibroblasts leads to the production of large amounts of collagen-based extracellular matrix.Therefore,to investigate the role of fibroblast fibrosis in the formation of pathological scar will provide a new idea for revealing the mechanism of pathological scar and biological therapy. OBJECTIVE:To investigate the effect of RAS-selective lethal small molecule 3(RSL3)on the fibrosis of human pathological scar fibroblasts. METHODS:Then cases of pathological scar tissue and normal skin tissue samples from the same individuals,provided by the Department of Burn Plastic Surgery,General Hospital of Ningxia Medical University,were collected.Fibroblasts of human pathological scar and human normal skin were extracted and used in the following experiments.The general condition of the pathological scar tissue and the normal skin tissue was detected by hematoxylin-eosin staining.The appearance of fibroblasts from pathological scar and normal skin were observed by inverted microscope.The fibroblasts were verified by immunofluorescence assay.The cells were treated with different concentrations of RSL3(1,3,5,7,9,11,13 μmol/L).The inhibitory concentration of RSL3 on fibroblasts was detected by cell counting kit-8.Control group(without treatment)and RSL3 intervention group(treated with 7 μmol/L RSL3 for 24 hours)were set up.The mRNA and protein expressions of glutathione peroxidase 4,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by Qrt-PCR and western blot,respectively.Level of malondialdehyde in cells was detected.The residual scratch area was measured by cell scratch test after 24 hours to calculate the percentage of residual scratch area. RESULTS AND CONCLUSION:The expression of glutathione peroxidase 4 in the pathological scar group was higher than that in the normal skin group(Mrna:t=3.252,P<0.01;protein:t=5.075,P<0.01).The expression of glutathione peroxidase 4 in the pathological scar fibroblast group was higher than that in the normal skin fibroblast group(Mrna:t=10.32,P<0.01;protein:t=26.22,P<0.01).Compared with the control group,the expression of glutathione peroxidase 4 was decreased(Mrna:t=2.798,P<0.05;protein:t=4.643,P<0.01),the content of malondialdehyde was increased(t=2.917,P<0.05),the expression of type Ⅰ collagen(Mrna:t=15.84,P<0.01;protein:t=4.610,P<0.01),type Ⅲ collagen(Mrna:t=28.86,P<0.01;protein:t=7.713,P<0.01)and α-smooth muscle actin(Mrna:t=2.671,P<0.05;protein:t=7.417,P<0.01)were decreased in the RSL3 intervention group.Compared with the control group,the migration ability was weakened in the RSL3 intervention group(t=14.06,P<0.01).To conclude,RSL3 can inhibit the expression of glutathione peroxidase 4 and then inhibit the ability of fibrosis and migration of pathological scar fibroblasts.

Objective: To evaluate the value of MRI-T₂WI texture analysis in the differentiation of atypical medulloblastoma and ependymoma of the fourth ventricle. Methods: Preoperative MRI data of 36 cases of fourth ventricle tumor (19 cases of medulloblastoma and 17 cases of ependymoma) confirmed by the Central Theater General Hospital of the Chinese People′s Liberation Army were retrospectively analyzed. Manually sketch areas of interest (ROI) were made using texture analysis software to get histogram parameters, including mean, median, standard deviation, skewness, kurtosis, maximum, minimum, heterogeneity, entropy, the 5th percentile (T₂WI_5th), the 10th percentile (T₂WI_10th), the 25th percentile (T₂WI_25th), the 50th percentile (T₂WI_50th), the 75th percentile(T₂WI_75th), the 95th percentile(T₂WI_95th). Independent sample t test or mann-whitney U test was used for statistical analysis of each parameter value between the two groups. Logistic regression analysis was used to perform multiparameter joint analysis on meaningful parameters. The area under the curve (AUC) was obtained by using the subject operating characteristic (ROC) curve to determine the threshold and diagnostic efficacy for distinguishing the two group. Results: The mean, median, T₂WI_5th, T₂WI_10th, T₂WI_25th, T₂WI_50th, T₂WI₇5th values of the medulloblastoma group were smaller than those of the ependymoma group, and difference was statistically significant (P<0.05). The T₂WI_5th had the best diagnostic performance. When the critical value was 1 343, the diagnostic specificity was 75.0% and the sensitivity was 92.9%. Logistic regression was used to predict the joint analysis of probability parameters. T₂WI_5th+T₂WI_10th had the best performance, and AUC was 0.928. When the critical value was 0.75, the diagnostic specificity was 75.0% and the sensitivity was 100.0%. Conclusions MRI-T₂WI texture analysis can provide quantitative information for preoperative diagnosis of atypical medulloblastoma and ependymoma and improve the accuracy of preoperative diagnosis, which has obvious clinical value.