Inducing the degradation of KRAS represents a novel strategy to combat cancers with KRAS mutation. In this study, we identify ubiquitin-specific protease 2 (USP2) as a novel deubiquitinating enzyme of KRAS in multiple myeloma (MM). Specifically, we demonstrate that gambogic acid (GA) forms a covalent bond with the cysteine 284 residue of USP2 through an allosteric pocket, inhibiting its deubiquitinating activity. Inactivation or knockdown of USP2 leads to the degradation of KRAS, resulting in the suppression of MM cell proliferation in vitro and in vivo. Conversely, overexpressing USP2 stabilizes KRAS and partially abrogates GA-induced apoptosis in MM cells. Furthermore, elevated USP2 levels may be associated with poorer prognoses in MM patients. These findings highlight the potential of the USP2/KRAS axis as a therapeutic target in MM, suggesting that strategically inducing KRAS degradation via USP2 inhibition could be a promising approach for treating cancers with KRAS mutations.

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.

Background Attack of primary acute angle-closure glaucoma (PAACG) often cause damage of retinal function.To measure the structural change after acute angle-closure glaucoma attack is of an important clinical significance.Flourier domain OCT(FD-OCT) is available.Objective This study was to measure and compare the thickness of peripapillary and macular retinal nerve fiber layer (RNFL) and macular volume in PAACG patients and normal subjects.Methods A case-controlled study was designed.Twenty-five eyes after acute angle-closure glaucoma attack and 25 fellow eyes were collected from April,2011 to February,2012 in Affiliated Eye Hospital of Wenzhou Medical College,and 25 eyes of normal subjects with the matched age and gender were enrolled at the same period.The thicknesses of RNFL and macula as well as the volumes of macula were measured by FD-OCT in all the eyes 2 weeks after acute angle-closure glaucoma attack.The outcomes were compared among the attacked eyes,fellow eyes and normal control eyes.Results The average thicknesses of RNFL were (125.72 ± 28.57),(108.36 ±9.31) and (106.10± 10.97)μm in the attacked eyes,fellow eyes and normal control eyes,showing a significant increase in the attacked eyes (all at P ＜ 0.05).The RNFL thickness values in the superior,interior and nasal quadrants at optical disc were significantly higher in the attacked eyes than those of the fellow eyes and normal control eyes (all at P＜0.05),but no significant differences were found in temporal and temporal-lower quadrants among the three groups (P=0.081,0.766).The thickness values of macular area were (283.72 ± 18.33),(280.28 ± 16.85) and (289.14±10.60)μm in the attacked eyes,fellow eyes and normal control eyes,with no significant difference among them (Fgroup =2.048,P =0.136),and fellow eyes were significantly reduced in comparison with the normal control group (P＜0.05),and those in the attacked eyes was not significantly reduced in comparison with the normal control group (P =0.224).The macular volumes were (5.589±0.355),(5.532±0.325) and (5.720±0.241)mm3in the attacked eyes,fellow eyes and normal eyes,without statistically significant difference among them (Fgup =1.027,P=0.363).The macular thickness values were lower in the outer race than those of the inner race and followed the pattern of nasal ＞superior＞ inferior＞temporal quadrants.At the temporal,superior,nasal and inferior quadrants of inner race,the temporal quadrant of the outer race,the central area,the macular thickness and volume values were declined in the attacked eyes and fellow eyes compared with the normal control eyes (all at P＜0.05).Conclusions The edema of RNFL is more serious than that of macula in the eyes within 2 weeks after acute angleclosure glaucoma attack.The retinas of macular zone are thinner,and the macular volumes are smaller in the attacked eyes and fellow eyes than those in the normal control eyes.