This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.

Objective:To recognize the characteristics of necrotizing fasciitis patients complicated with sepsis and summarize the experience the treatment.Methods:A retrospective study was conducted. The clinical data of 57 patients with necrotizing fasciitis complicated with sepsis admitted to Guangdong Provincial People's Hospital from July 2009 to December 2019 was analyzed by collecting such factors as gender, age, complications, infection sites, pathogens, surgery information, treatment options and outcome. The patients were divided into debridement group ( n = 14) and control group ( n = 43) according to whether the debridement was completed within 48 hours of admission, and the mortality during hospitalization between the two groups was compared. A telephone follow-up had been done to record the long-term outcome of these patients. Results:Among 57 patients with necrotizing fasciitis complicated with sepsis, there were 43 males and 14 females with the average age of (57.9±12.1) years old. Most of the underlying diseases were diabetes mellitus (70.17%), other diseases included hypertension (8.77%), tumor chemotherapy (7.02%), liver disease (hepatitis, cirrhosis, 7.02%), coronary artery heart disease (3.51%), systemic lupus erythematosus (3.51%), etc. Most of the infection site was lower limbs (71.93%). There were 78 pathogens cultured in 57 patients, in which 52 were non-drug resistant bacteria (66.67%), and 26 were drug resistant bacteria (33.33%). There were 40 Gram positive (G +) bacteria (51.28%), 29 Gram negative (G -) bacteria (37.18%), 8 fungi (10.26%) and 1 mixed bacteria (1.28%). Finally, of 57 patients, 46 patients were cured, and 11 patients died with hospital mortality of 19.30%. Among 57 patients, the hospital mortality in the debridement group was significantly lower than that in the control group [0% (0/14) vs. 25.58% (11/43), P < 0.05]. Among the 46 cured patients, 11 had accepted amputations, accounting for 23.91%. In December 2020, 43 patients who were cured (3 patients were lost to follow-up) were followed up by telephone. Twenty-three patients were completely self-care, 9 patients were partly self-care, 8 patients were completely unable to take care of themselves, and 3 patients died. Conclusions:Necrotizing fasciitis with sepsis mostly occurs in people with weakened immunity, and has a high mortality and disability rate. Early identification and active surgical debridement may be the key to improve the treatment effect.

Objective To investigate the potential protective effects of valproic acid (VPA) on gut barrier function after major burn injury in rats and its mechanism.Methods Forty male Sprague-Dawley (SD) rats were divided into sham + normal saline (NS),sham + VPA,scald + NS,and scald + VPA groups,with 10 rats in each group.Rat with 55％ total body surface area (TBSA) third-degree severe-bums model was reproduced by immersing into 80 ℃ water,and the rats in sham groups were given sham-bums by immersing into 37 ℃ water.The rats after severebums were immediately treated with 0.25 mL of 300 mg/kg VPA or NS by subcutaneous injection.Rats were sacrificed at 2 hours and 6 hours after injury,and abdominal aortic blood and ileal tissue were harvested.The levels of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay (ELISA).The intestinal permeability was evaluated by fluorescein isothiocyanate-dextran (FITC-dextran) determination.The histomorphological changes in gut barrier were evaluated by Chiu grading system.Levels of acetylated lysine at the ninth position of histone 3 protein (Ac-H3K9),hypoxia-inducible factor 1α (HIF-1α),zona occludens 1 (ZO-1) and myosin light chain kinase (MLCK) were determined by immunofluorescence staining and Western Blot.Results Compared with sham + NS group,rats in scald + NS group showed intestinal mucosal damage 2 hours after bum injury,as well as increased mucosal permeability,protein expression levels of HIF-1 α,VEGF,MLCK,and lowered levels of AC-H3K9 and ZO-1.These changes were much more prominent at 6 hours after injury.VPA treatment significantly attenuated the bum-induced intestinal damage.Compared with scald + NS group,the protective effects in scald + VPA group was not evident at 2 hours after injury;however,intestinal damage was much less severe at 6 hours after injury (Chiu score:2.03 ± 0.27 vs.3.12 ± 0.15),intestinal permeability was significantly decreased [FITC-dextran (μg/L):709 ± 76 vs.1138 ± 75],histone acetylation was enhanced [Ac-H3K9 (gray value):1.55 ± 0.12 vs.0.48±0.12],ZO-1 degradation was significantly inhibited (gray value:0.69 ± 0.12 vs.0.43 ± 0.16),the protein expression levels of VEGF and MLCK were significantly down-regulated [VEGF (ng/mg):51.7±3.7 vs.71.2±4.3,MLCK (gray value):1.98±0.20 vs.2.80±0.24],while the HIF-1 α protein expression levels were significantly reduced at both 2 hours and 6 hours after injury (gray value:2.50±0.39 vs.3.88±0.42 at 2 hours,1.83±0.42 vs.4.42±0.41 at 6 hours,all P ＜ 0.05).Conclusions Severe bum injury can induce histone deacetylation,ZO-1 degradation and intestinal barrier dysfunction.VPA can improve the levels of histone acetylation and ZO-1,and protect intestinal epithelial barrier function.These may probably be mediated through inhibiting HIF-1α and its downstream gene VEGF and MLCK.

This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.
Active Transport, Cell Nucleus ; physiology ; Breast Neoplasms ; metabolism ; physiopathology ; Caveolin 1 ; metabolism ; Cell Line, Tumor ; Female ; Humans ; Protein Transport ; physiology ; Radiation ; Receptor, ErbB-2 ; metabolism

Objective To analyze the setup errors by cone-beam computed tomography (CBCT) for breast cancer patients who were immobilized with neck and breast thermoplastic mask and received intensity modulated radiation therapy (IMRT), and to calculate the external margins from the clinical target volume (CTV) to the planning target volume (PTV) (MPTV) of tumors. Methods Twenty-five breast cancer patients who were immobilized with neck and breast thermoplastic mask and received IMRT in the Oncology Department of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from November 2016 to June 2017 were enrolled. The position of the patients were verified by CBCT before treatment . The linear and rotation errors of the X, Y and Z axes were analyzed by online bone registration. The systematic errors (Σ) and random errors (σ) of the patients were also calculated, and then the margins from CTV to PTV margins were calculated based on MPTV=2.5Σ+0.7σ. 25 patients'height, weight, body mass index (BMI) and the maximum diameters of CTV in the lateral, longitudinal and vertical directions were recorded, and the relation between the setup errors and the above mentioned was analyzed by using Spearman method. Results A total of 174 CBCT scans for 25 breast cancer patients were completed. The group Σ were 1.40 mm, 1.50 mm and 1.20 mm, and rotation errors were 0.9°, 0.7° and 0.8° at the X, Y and Z axes, respectively. The group σ were 2.20 mm, 3.00 mm and 1.40 mm, and rotation errors were 0.7°, 0.6° and 0.7° at the X, Y and Z axes, respectively. MPTVwas recommended as 4.90 mm, 6.00 mm and 3.90 mm at the X, Y and Z axes, respectively. There was no correlation between the height, weight, BMI of the patients and the setup errors (all P ＞ 0.05). However, there was a significant correlation between the maximum lateral, longitudinal diameters of the CTV and the setup errors (rs= 0.406, P= 0.044; rs= 0.512, P= 0.009). Conclusions The neck and breast thermoplastic mask can improve the diagnostic accuracy of radiotherapy in breast cancer patients. The data of setup errors verified by CBCT can provide meaningful references for the setting of MPTV.

Objective To investigate the accuracy and application value of optical surface monitoring system in intensity modulated radiotherapy for thoracic tumors patients.Methods Twenty-eight patients with thoracic tumors were included.During each treatment fraction,the patients were immobilized with body surface markers and laser lamps.The surface images obtained by the optical surface monitoring system were registered with the reference images and recorded during the CBCT scan.The translation and rotation errors of x (left-right),y (craniocaudal) and z (anterior-posterior) axes were recorded.After scanning,the CBCT images were registered with the planned CT images and the translation and rotation errors of x,y and z axes were recorded.The setup errors of these two image systems were analyzed and corrected before each treatment.The correlation between the two sets of setup errors were analyzed with Pearson test,and systematic error (∑) and random error (σy) were also calculated.The consistency of the two image systems was evaluated with the Bland-Altman method and the 95％ limits of agreement were calculated.Results There was a good correlation between these two groups,and the correlation coefficients were 0.79,0.62,and 0.53 in x,y and z axes,respectively.The ∑/σr of the optical surface monitoring system were 0.7 mm/1.5 mm,0.9 mm/1.8 mm and 0.9 mm/1.5 mm in x,y and z axes,respectively.The ∑/σ of CBCT were 0.8 mm/1.6 mm,1.3 mm/1.9 mm and 0.7 mm/1.5 mm in x,y and z axes,respectively.The 95％ limits of agreement of translations direction were (-2.0-2.3),(-3.4-3.6) and (-3.3-2.4) mm,and the 95％ limits of agreement of rotation direction were (-2.0 to 1.6)°,(-2.0 to 1.4)° and (-1.6 to 1.6)° inx,y and z axes,respectively.Conclusions The optical surface monitoring system is an effective image guide tool,which can quickly and accurately verify the patient's position and improve the position accuracy.It can be applied for positioning in the intensity modulated radiation treatments for the thoracic tumor patients.

Objective:To explore the clinical application value of Bodyfix fixation device in stereotactic body radiotherapy (SBRT) for elderly patients with lung cancer.Methods:The clinical data of 63 elderly patients with lung cancer who received SBRT in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January to October 2021 were retrospectively analyzed. According to different fixation methods, the patients were divided into Bodyfix combined with vacuum bag fixation device group (Bodyfix group, 20 cases) and 4D respiratory gating technology combined with vacuum bag fixation device group (vacuum bag group, 43 cases). Cone beam CT (CBCT) was used for position verification before each treatment, linear and rotational errors in the horizontal (X), head-to-foot (Y), front-to-back (Z) directions were recorded.Results:The linear errors of Bodyfix group in the X, Y and Z directions were 1.7 mm (1.3 mm, 3.0 mm), 4.6 mm (4.3 mm, 5.3 mm) and 1.3 mm (0.8 mm, 2.8 mm), and the rotational errors were (0.46±0.04)°, (-0.48±0.05)° and 0.64°(0.38°, 1.07°); the linear errors of vacuum bag group in the X, Y and Z directions were 2.1 mm (1.6 mm, 3.3 mm), 2.8 mm (1.8 mm, 3.7 mm) and 3.0 mm (2.3 mm, 3.8 mm), and the rotational errors were (0.69±0.04)°, (-0.70±0.04)° and 0.64° (0.42°, 0.86°). The differences in linear errors in the Y and Z directions and rotational errors in the X and Y directions between the two groups were statistically significant ( P values were <0.001, <0.001, 0.003 and 0.007). Conclusions:Compared with the 4D respiratory gating technology, the Bodyfix fixation device has smaller rotational errors in the X and Y directions and linear errors in the Z direction. It can be used as an effective method of postural fixation for SBRT in elderly patients with lung cancer.

OBJECTIVETo analyze the development of liver damage and reactivation of hepatitis B virus (HBV) during the treatment of extremely severe burn injury in HBsAg positive patients, in order to provide reference for prevention and treatment of liver damage in patients with HBV infection after extremely severe burn.

METHODSMedical records of 54 HBsAg positive patients after extremely severe burn injury admitted from January 2004 to December 2014 were retrospectively analyzed. Development of liver damage and HBV reactivation of these patients during the treatment were analyzed according to the classification of their gender, results of hepatitis B e antigen (HBeAg) and HBV DNA examinations on admission, and development of sepsis in the process of treatment. Data were processed with chi-square test.

RESULTS(1) The incidence of liver damage in the process of treatment of these patients was 85.2% (46/54). Among all the patients, the proportion of liver damage was 35/38 in male, which was significantly higher than that in female (11/16, χ² = 4.867, P<0.05). Liver damage was found in all of 26 patients who were HBeAg positive on admission, 34 patients who were HBV DNA positive on admission, and 36 patients who developed sepsis in the process of treatment; the proportions were significantly higher than those in patients who were HBeAg negative on admission (20/28), patients who were HBV DNA negative on admission (12/20), and patients who did not develop sepsis in the process of treatment (10/18), with χ² values respectively 11.801, 18.384, and 20.574, P values below 0.01. (2) The incidence of HBV reactivation in these patients was 29.6% (16/54). Among all the patients, the proportion of HBV reactivation was 13/38 in male and 3/16 in female, with no statistically significant difference between them (χ² = 0.656, P>0.05). The proportions of HBV reactivation in patients who were HBeAg positive on admission, patients who were HBV DNA positive on admission, and patients who developed sepsis in the process of treatment were respectively 13/26, 16/34, and 15/36, and they were significantly higher than those in patients who were HBeAg negative on admission (3/28), patients who were HBV DNA negative on admission (0/20), and patients who did not develop sepsis in the process of treatment (1/18), with χ² values respectively 9.979, 18.615, and 5.873, P<0.05 or P<0.01.

CONCLUSIONSPatients who are HBsAg positive, HBeAg positive, HBV DNA positive on admission, and develop sepsis in the process of treatment of extremely severe burn injury are more likely to develop liver damage and HBV reactivation. It is necessary to dynamically monitor the changes in HBV DNA and liver function, in order to identity the reactivation of virus.

Alanine Transaminase ; blood ; Burns ; complications ; drug therapy ; Chemical and Drug Induced Liver Injury ; DNA, Viral ; Female ; Hepatitis Antibodies ; blood ; Hepatitis B ; drug therapy ; epidemiology ; virology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B virus ; drug effects ; immunology ; isolation & purification ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Humans ; Incidence ; Liver ; pathology ; Male ; Retrospective Studies