Shengmai Yin (SMY) is a Chinese herbal decoction that effectively alleviates the side effects of radio-therapy in various cancers and helps achieve radiotherapy's clinical efficacy.In this study,we explored the interaction mechanism among SMY,DNA methylation,and nasopharyngeal carcinoma (NPC).We identified differences in DNA methylation levels in NPC CNE-2 cells and its radioresistant cells (CNE-2R)using the methylated DNA immunoprecipitation array and found that CNE-2R cells showed genome-wide changes in methylation status towards a state of hypomethylation.SMY may restore its original DNA methylation status,and thus,enhance radiosensitivity.Furthermore,we confirmed that the dif-ferential gene Tenascin-C (TNC) was overexpressed in CNE-2R cells and that SMY downregulated TNC expression.This downregulation of TNC inhibited NPC cell radiation resistance,migration,and invasion.Furthermore,we found that TNC was hypomethylated in CNE-2R cells and partially restored to a hypermethylated state after SMY intervention.DNA methyltransferases 3a may be the key protein in DNA methylation of TNC.

Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.

Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.

Objective To investigate the effect of moxibustion or acupuncture combined with prokinetic drugs (metoclopramide injection) on the success rate of placing the nasal and intestinal tubes in critically ill patients at bedside.Methods A total of 107 patients underwent enteral nutrition from January 2015 to August 2017 in the Intensive Care Unit (ICU) of Wenzhou Central Hospital in Zhejiang Province were recruited and given bedside manual placement of spiral nasointestinal tube. According to whether the acupuncture or moxibustion combined with the use of the prokinetic drugs in the process of placing the tube, the cases can be divided into three groups: simple prokinetic drugs group (n=23), moxibustion combined with prokinetic drugs group (n=22), acupuncture and moxibustion combined with prokinetic drugs group (n=62). The success rate of catheterization in three groups was analyzed and compared statistically.Results After 72 hours of catheterization, bedside X-ray abdominal radiography showed that the end of the catheter was located behind the pylorus in 81 cases out of the 107 patients. The success rate of catheterization was 75.70％. The unsuccessful patients were given gastroscopy guided nasointestinal tube. The success rate of catheterization was 30.43％ in simple prokinetic drugs group, 63.64％ in moxibustion combined with prokinetic drugs group, and 96.77％ in acupuncture and moxibustion combined with prokinetic drugs group. Comparing the success rate of the catheterization, the difference was statistically significant (χ2=49.75,P<0.01).Conclusions The combination of moxibustion and acupuncture can significantly increasing the success rate of placing the spiral nasointestinal tube, and provide practical benefits for the critically ill patients. This method should be popularized in clinic.

Objective: To observe the effects of axial vascular network flap of scalp or anterolateral thigh perforator flap with fascia lata on repairing defects after radical resection of scalp carcinoma in patients. Methods: From February 2006 to December 2015, twenty-one patients with scalp carcinoma were admitted to our hospital, and the carcinoma invaded external lamina or full-thickness of skull and dura mater. After perfect preoperative examination, carcinoma and scalp tissue in 3 to 5 cm from the edge of carcinoma, external lamina or full-thickness of skull and invaded dura mater were resected and sentinel lymph nodes around carcinoma were cleaned in 3 to 4 days after admission. The postoperative defects with size reached from 11 cm×8 cm to 22 cm×18 cm. The flap transplantation was performed at the same time when quick frozen pathological examination results of resected scalp carcinoma margin tissue, skull, dura mater margin and basal tissue, and sentinel lymph nodes showed completely negative. Defects in 3 elderly patients were repaired by single or multiple axial scalp vascular network flaps, with the resected flaps size ranged from 12 cm×7 cm to 19 cm×14 cm. Defects in the other 18 patients were repaired by anterolateral thigh perforator flaps with fascia lata, with the resected flaps size ranged from 13 cm×10 cm to 23 cm×19 cm and the resected fascia lata size ranged from 8 cm×7 cm to 10 cm×10 cm. The head donor site of flap was repaired by medium thickness skin of head and back; the thigh donor site of flap was repaired by medium thickness skin of thigh on the same side. All patients gave up postoperative radiotherapy, chemotherapy, and other follow-up treatments. Results: After operation, the flap and skin in all patients survived completely, with no vascular crisis or other condition. During the follow-up for 6 months to 9 years, all patients showed good appearance except for baldness in operation area of head, with no obvious malformation in head donor site of flap and skin, no swollen external hernia in the brain tissue, and no local recurrence or distant metastasis of carcinoma. The appearance of thigh donor site of flap and skin was good, with normal muscle strength and movement of lower limbs. Conclusions Patients with scalp carcinoma were performed with radical resection of carcinoma, and axial vascular network flap of scalp or anterolateral thigh perforator flap with fascia lata were applied to repair the postoperative defects, with good appearance of head operation area and no local recurrence or distant metastasis of carcinoma.

Objective To establish an HPLC method for the simultaneous determination of the 6 aconitum alkaloids in compoundTengwu Ointment.Methods Chromatographic column was Aglient XDB-C18 column (4.6 mm × 250 mm, 5μm), with the mobile phase A of acetonitrile- tetrahydrofuran (25∶8), phase B was 0.1 mol/L ammonium acetate solution (0.5 mL acetic acid added to the 1000 mL) with gradient elution;the flow rate was 1 mL/min;the detection wavelength was set at 235 nm;the column temperature was 25℃.Results Aconitine was in the good linear range of 0.072-0.648μg (r=0.999 5), with the average recovery of 99.29%, RSD=1.25%. Mesaconitine was in the good linear range of 0.062-0.648μg (r=0.999 5), with the average recovery of 99.12%, RSD=0.85%. Hypaconitine was in the good linear range of 0.064-0.384μg (r=0.999 8), with the average recovery of 99.57%, RSD=1.07%. Benzoylaconine was in the good linear range of 0.056-0.672μg (r=0.999 2), with the average recovery of 98.11%, RSD=0.61%. Benzoylmesaconine was in the good linear range of 0.055-0.993μg (r=0.999 9), with the average recovery of 99.27%, RSD=1.10%. Benzoylhypaconine was in the good linear range of 0.078-0.702μg (r=0.999 8), with the average recovery of 99.08%, RSD=1.38%.Conclusion This method is simple, sensitive, repeatable and accurate, which can be used for determination of aconitum alkaloids in compoundTengwu Ointment.

Objective To optimize the matrix formula of propranolol hydrochloride gels. Methods On the basis of single factor experiment, central composite design-response surface method was used to optimize the formula. Addition levels of glycerol (A), PEG-400 (B) and HPMC (C) were evaluated as the independent variables.Eight-hour accumulative penetration amount per unit area measured by HPLC was used as the index. Quadratic polynomial was used to estimate the relationship between the index and the independent variables, and to delineate response surface and overlay contour plots in order to select the optimal formulations.Finally, predicted responses were verified. Results The optimized formula consisted of 18.53% glycerol, 8.54% PEG-400 and 2.35% HPMC.The quadratic polynomial regression model of 8-h accumulative penetration amount per unit area:R1=-7 415. 69+306. 10A+167. 47B+4 820. 59C-8. 26A2-9. 81B2-1 025. 75C2 , and the cumulative transmittance was 49.6%. Conclusion A credible model is established by using central composite design-response surface method and the formula of propranolol hydrochloride gels is optimized and the gel is stable and controllable.

Objective To investigate the biological effects of 125I seeds and 60Co γ-rays on the non-small cell lung cancer cells A549 and the normal bronchial epithelium cells BEAS-2B.Methods A549 and BEAS-2B cells were irradiated with 125I seeds and 60Co γ-rays.The survival fraction was detected by colony formation assay.The cell cycle and cell apoptotic ratio were detected by flow cytometry.The expression of cell apoptotic related proteins was examined by western blot.Results After irradiation with different doses,the survival of A549 cells irradiated with 125I seeds was lower than that irradiated with γ-rays (t =6.06,9.42,4.90,P ＜0.05).After irradiation with 4 Gy of 125I seeds and 60Co γ-rays,the G1 phase percentages of A549 cells were 70.67％ ± 1.49％ and 59.59％ ± 0.71％ (t =10.77,P ＜ 0.05),and the apoptotic ratios of A549 cells were 18.09％ ±0.73％ and 9.81％ ±0.16％ (t =19.40,P＜ 0.05),respectively.125I seeds irradiation remarkably up-regulated the expressions of Bax and cleaved Caspase-3 proteins,down-regulated the expression of Bcl-2 proteins compared with 60Co γ-rays irradiation on A549 cells.However,the apoptotic ratio and the expressions of apoptosis-related proteins in BEAS-2B cells had little difference between two types of radiation.Conclusions The anti-proliferative effect of 125I seeds irradiation on A549 cell is more remarkably than that of 60Co γ-rays.The imbalance of Bcl-2/Bax ratio and the eventually activation of Caspase-3 proteins may play an important role in the anti-proliferative effect induced by the continuous low dose radiation of 125I-seeds.

Objective To determine the eugenol in Dijin spray by GC-MS.Methods The chromato-graphic column was DB-5 fused silica capillary column.Eugenol was determined by program temperature risen method:the initial temperature was 60 ℃ for 1 min,then raised to 200 ℃ at the rate of 10 ℃/min and kept for 5 min,and at last raised to 250 ℃ at 15 ℃/min and kept for 5 min.Electron energy was 70ev.The ions of 164,149,and 77 were selected for eugenol detection.Results The calibration curve of eugenol was linear in the range of0.116～1.16 mg/ml (r=0.999 4) with the average recovery of 97.84％ (RSD=0.47％).Conclusion The method was quick,accurate and repeatable.It was suitable for the quality control of Dijin spray.

Aim To study effects of loganin and morro-niside on aging astrocytes induced by D-galactose. Methods Cortex astrocytes of newly born rats were cultured in vivo and indentified by immunofluorescence method.Firstly,appropriate D-galactose concentration was selected and effects of loganin and morroniside on proliferation activity of aging astrocytes induced by D-galactose were determined by MTT test.Then SOD, MDA were taken as indicators to study the effects of loganin and morroniside on aging astrocytes induced by D-galactose.Thirdly,growth factors like gliar cell line-derived neurotrophic factor (GDNF),basic fibro-blast growth factor (bFGF)tested by ELISA method and expression of Bax,caspase-3,phospho-extracellu-lar signal-regulated kinese (p-ERK1 /2),phospho-mi-togen-actived protein kinase /extracellular signal-regu-lated kinase kinase (p-MEK1 /2)were taken as indi-cators to discuss the potential protection mechanism. Results Loganin and morroniside exerted certain effects on proliferation of aging astrocytes induced by D-galactose,improving SOD,GDNF,bFGF release and lowering MDA release significantly (P <0.05 ). The expressions of Bax and caspase-3 proteins had no difference between model group and loganin group, morroniside group.While the expressions of p-ERK1 /2,p-MEK1 /2 of loganin group,morroniside group were improved significantly compared with model group.Conclusion Loganin and morroniside have protective effects on aging astrocytes induced by D-ga-lactose,and increase the proliferation ability.Protec-ting their antioxidant systems and improving the expres-sion of p-ERK1 /2,p-MEK1 /2 proteins are possible mechanisms.