Objective:To develop a novel magnetic composite nanoparticle with excellent targeting and biocompatibility and verify synergistic magnetic hyperthermia/radiotherapy efficacy in HeLa cells.Methods:Bovine serum albumin(BSA)was used to modify ferroferric oxide(Fe 3O 4)nanoparticles, and then L-selenocystine and folic acid(FA)were coupled on the surface of BSA by carbodiimide method to synthesize magnetic composite nanoparticles Fe 3O 4@BSA@SeCyc2@FA.The as-prepared nanoparticles were characterized to determine the structure, composition, and properties of the nanoparticles to support their biological applications.By evaluating the magnetothermal properties of the as-prepared nanoparticles, analyzing the internalization effect of HeLa cells on the particles and using CellTiter and reactive oxygen species(ROS)kits to detect the cell viability and ROS production of different experimental groups, respectively, the killing and inhibiting effects on HeLa cells were evaluated when hyperthermia was combined with radiotherapy. Results:The average particle size of the Fe 3O 4@BSA@SeCyc2@FA nanoparticles was(19.31 ± 4.84)nm, and the zeta potential was -25.4 mV(pH=7), which indicated that the as-prepared nanoparticles maintained acceptable dispersion and stability for biological experiments.By BSA and FA characteristic absorption peaks combined with the measured content of Se(10.89 μM), L-selenocystine and FA were successfully modified on the surface of the composite nanoparticles.The saturation magnetization value of the Fe 3O 4@BSA@SeCyc2@FA nanoparticles was 47.2 emu/g.The as-prepared nanoparticles maintained the specific absorption rate(SAR)value of 125.4 W/g at the alternating magnetic field of 518 kHz/16 kAm -1, indicating excellent heat generation efficiency(the temperature level ΔT=7 ℃ within 15 min).No significant toxicity was found for HUVEC cells even within the concentration range from 0 to 200 μg/ml and inhibition of cell viability of the HeLa was shown.When magnetic hyperthermia combined with high-energy X-ray, the cell viability of the HeLa decreased significantly to 54.7%, and the intracellular ROS production increased by 108.2% compared with the control group, which showed that the as-prepared nanoparticles significantly enhanced the radiosensitivity of HeLa cells, and the synergistic effect of magnetic hyperthermia combined with radiotherapy was more effective in inhibiting and killing them. Conclusions:The as-prepared nanoparticles show great promise for use as a multifunctional nanoplatform for the synergistic magnetic hyperthermia/radiotherapy to overcome the limitations of monotherapy for cancer treatment.

This article reported a case of pyruvate dehydrogenase E1-α deficiency suggested by abnormal brain development during prenatal ultrasound imaging. Prenatal ultrasound revealed a mild enlargement of bilateral cerebral ventricles and the possibility of intracranial hemorrhage in the fetus at 25 +1 weeks of gestation. MRI showed the fetus with absent corpus callosum, enlarged bilateral cerebral ventricles and paraventricular cysts. After genetic counseling and careful consideration, the couple opted for pregnancy termination. To clarify the cause of the disease, whole-exome sequencing was performed on the fetal skin to detect possible variants, and which revealed a frameshift mutation c.924_930dup(p.R311Gfs*5) in exon 10 of the PDHA1 gene. Sanger sequencing confirmed the mutation was a de novo pathogenic variant, indicating that the fetus was affected by pyruvate dehydrogenase E1-α deficiency.

Objective:To investigate the role of growth arrest specific protein 6 (Gas6) in regulating macrophage polarization in wound healing.Methods:Clean male B6 mice were randomly(random number) divided into the normal group, skin defect group, skin defect group + normal saline group (PBS group), skin defect + Gas6 (1 μg) group, skin defect + Gas6 (5 μg) group, and skin defect + Gas6 (10 μg) group. Ten mice in each group were used to observe the healing of skin wounds. Macrophages were isolated from the wound tissues of the remaining 6 mice on the fifth day after modeling. The levels of IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA), the mRNA expression levels of arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) were detected by RT-PCR, and flow cytometry was used to detect the expression of M1 marker CD197, M2 marker CD163 and F4/80. HE staining was used to detect the pathological changes of skin wounds. Masson staining was used to analyze the granulation tissue and collagen deposition.Results:Scab began to form on the surface of the wound on the third day after the skin defect model was established. The wound area of the Gas6 treatment group was smaller than that of the PBS group, and the wound healing was better than that of the PBS group. Compared with the normal group, the proportion of CD197 in macrophages of the skin defect group was significantly increased ( P=0.00 49), the proportion of CD163 and F4/80 double positive was significantly decreased ( P=0.00 86), the level of IL-6 was significantly increased ( P=0.00 13), the level of IL-10 was significantly increased ( P=0.00 14), the level of iNOS mRNA was significantly increased ( P=0.00 8), and Arg-1 was significantly increased in the skin defect group The mRNA level was significantly decreased ( P=0.01 21), and the inflammatory infiltration was aggravated. Compared with the PBS group, the proportion of CD197 in the Gas6 treatment group was significantly decreased ( P=0.00 0), the double positive rates of CD163 and F4/80 were significantly increased ( P = 0.00 0), the level of IL-6 was significantly decreased (P = 0.00 0), the level of IL-10 was significantly increased ( P=0.00 03), the level of iNOS mRNA was significantly decreased ( P=0.00 18), the level of Arg-1 mRNA was significantly increased ( P=0.00 1), and the number of inflammatory cells and the number of collagen fibers were increased. Conclusions:Gas6 can promote the transformation of macrophages from M1 to M2 in mice with skin defect, which is beneficial to the wound healing of skin defect.

Objective: To explore the clinical effects of split-thickness scalp and allogenic acellular dermal matrix (ADM) in repairing deep wounds of hands in patients with extremely extensive burns. Methods: Six patients with extremely extensive burns complicated by deep burn in hands were admitted to our department from December 2014 to December 2017, including 4 males and 2 females, aged 21 to 58 years. Their total burn areas were from 85% to 95% total body surface area (TBSA), and the sum of deep-partial thickness and full-thickness wounds was larger than 50% TBSA. Under general anesthesia, deep wounds of 10 dorsal hands were repaired by combined transplantation of split-thickness scalp and allogenic ADM in 4 to 6 weeks after injury. The skin grafting range was beyond the metacarpophalangeal joint, including partial dorsal digital deep wounds. Survival of skin grafts on dorsal hands, follow-up of hand shape and functional recovery were observed and recorded. Results: Six patients were successfully treated. There was no infection on the dorsal deep wounds of 10 hands after surgery, and the grafts survived well. During follow-up of 1 to 2 years after operation, there were no recurrent tension blisters or dorsal extension deformity of the metacarpophalangeal joints in 10 hands, the shape and function of hands recovered well, and hand Carroll scores were from 90 to 99 points, with functional levels of Ⅴ and Ⅵ. Conclusions The combined transplantation of split-thickness scalp and allogenic ADM is a good method for repairing deep hand wounds in patients with extremely extensive burns, which can alleviate the cicatrix hyperplasia and contracture of healed hand wounds, and improve the shape and function of hands.

Objective: To investigate the clinicopathological features of indolent T-cell lymphoproliferative disorder of the gastrointestinal tract. Methods: Five cases of indolent T-cell lymphoproliferative disorder of the gastrointestinal tract from the Affiliated Hospital of Qingdao University from 2016 to 2019 were retrospectively reviewed. The clinical and pathological parameters were analyzed by combining clinical data and reviewing the available literature of 35 cases (34 cases abroad and 1 case in China). Results: There were 4 males and 1 female with a median age of 47 years (18-66 years). All patients had abdominal pain and constitutional symptoms including diarrhea, emaciation, intermittent mucous stool or oral and epiglottic ulcers. Endoscopic manifestations included multiple punctate congestion, erosion and ulcer at the terminal ileum and colorectum. Two cases had congestion and erosion of antrum and angle of stomach, and the lesions did not fuse and form tumors. Histologically, the lamina propria was expanded by a dense, medium to small lymphocyte infiltration, which was monomorphic, with slightly irregular nuclei without prominent nucleolus or lymphoepithelial lesions. There were admixed small amount of plasma cells and eosinophils. In 4 cases, immunohistochemistry showed the lesional cells were positive for CD3, CD8, TIA1, and negative for CD4, CD56, granzyme B and Ki-67 index was ≤10%. In situ hybridization showed that EBER was negative and clonal TCR gene rearrangement was detected. One consultation case was CD3⁺, CD5^- and Ki-67 index of 10%, although other indicators were not done. All five patients were treated with symptomatic support. In follow-up observation for 2 to 25 months, all patients were alive with the disease. Conclusions Indolent T-cell lymphoproliferative disorder of the gastrointestinal tract is a newly classified monoclonal T-cell proliferative disease, with low incidence, clinical inertia and long-term survival. It has unique clinicopathological features but pathologically it is easily misdiagnosed as inflammatory bowel disease or T-cell lymphoma. Correct diagnosis is of great important clinical significance.

Objective To investigate the clinicopathological features of indolent T?cell lymphoproliferative disorder of the gastrointestinal tract. Methods Five cases of indolent T?cell lymphoproliferative disorder of the gastrointestinal tract from the Affiliated Hospital of Qingdao University from 2016 to 2019 were retrospectively reviewed. The clinical and pathological parameters were analyzed by combining clinical data and reviewing the available literature of 35 cases (34 cases abroad and 1 case in China). Results There were 4 males and 1 female with a median age of 47 years (18-66 years). All patients had abdominal pain and constitutional symptoms including diarrhea, emaciation, intermittent mucous stool or oral and epiglottic ulcers. Endoscopic manifestations included multiple punctate congestion, erosion and ulcer at the terminal ileum and colorectum. Two cases had congestion and erosion of antrum and angle of stomach, and the lesions did not fuse and form tumors. Histologically, the lamina propria was expanded by a dense, medium to small lymphocyte infiltration, which was monomorphic, with slightly irregular nuclei without prominent nucleolus or lymphoepithelial lesions. There were admixed small amount of plasma cells and eosinophils. In 4 cases, immunohistochemistry showed the lesional cells were positive for CD3, CD8, TIA1, and negative for CD4, CD56, granzyme B and Ki?67 index was ≤10%. In situ hybridization showed that EBER was negative and clonal TCR gene rearrangement was detected. One consultation case was CD3+, CD5-and Ki?67 index of 10%, although other indicators were not done. All five patients were treated with symptomatic support. In follow?up observation for 2 to 25 months, all patients were alive with the disease. Conclusions Indolent T?cell lymphoproliferative disorder of the gastrointestinal tract is a newly classified monoclonal T?cell proliferative disease, with low incidence, clinical inertia and long?term survival. It has unique clinicopathological features but pathologically it is easily misdiagnosed as inflammatory bowel disease or T?cell lymphoma. Correct diagnosis is of great important clinical significance.

Objective To evaluate the clinical efficacy and adverse events of intensity-modulated radiotherapy ( IMRT ) in the treatment of intermediate risk localized prostate cancer, and analyze the significance of prostate-specific antigen ( PSA) level changes. Methods Clinical data of 66 patients with intermediate risk localized prostate cancer admitted to our hospital between 2007 and 2018 were retrospectively analyzed. Sixty patients were treated with endocrine therapy before radiotherapy. The radiation field covered the pelvic lymph node drainage area in 6 cases. Forty-seven patients received image-guided radiotherapy ( IGRT) . The median dose in the prostate and seminal vesicle was 78 Gy and 48 Gy in the pelvic lymph node drainage area. The survival rate was calculated using the Kaplan-Meier method. Results The median age was 77 years. The median follow-up time was 71. 3 months. The 5-year sample size was 47. The 3-and 5-year overall survival (OS) was 98％ and 90％.The 3-and 5-year cancer-specific survival (CSS) was 100％ and 93％.The 3-and 5-year biochemical relapse-free survival was 97％ and 86％. The mean time of PSA declining to the nadir was 5. 83 months. The median level of PSA nadir was 0. 06 ng/ml after IMRT. The incidence of grade I andⅡearly adverse events in the urinary system was 38％ and 6％. The incidence of grade I andⅡearly adverse events in the gastrointestinal system was 21％ and 3％. The incidence of grade I andⅡadvanced-stage adverse events in the urinary system was 9％ and 2％. The incidence of grade I advanced-stage adverse events in the gastrointestinal system was 5％. Conclusions IMRT yields high clinical efficacy in the treatment of intermediate risk localized prostate cancer with a low risk of adverse events in the early and advanced stage. The monitoring of PSA after IMRT contributes to the assessment of clinical prognosis.

Objective To analyze the data from intensity-modulated radiotherapy ( IMRT) for prostate cancer guided by kilovoltage cone-beam computed tomography (CBCT), and to provide a clinical basis for selecting the optimal image registration method and reasonable target volume margins.Methods A total of 16 patients with prostate cancer who received radical IMRT were enrolled, and CBCT for online position verification was performed 214 times.The images were obtained after conventional skin marking and laser alignment, and automatic registration, bone registration, soft tissue registration, and manual registration were performed for CBCT images and planned CT images.The differences between these four registration methods were evaluated, and the margins for extending clinical target volume into planning target volume (PTV) were calculated.Results The setup errors in left-right, anterior-posterior, and cranial-caudal directions for automatic registration, bone registration, soft tissue registration, and manual registration were-0.6±2.8 mm/-0.6±4.5 mm/-0.6±3.8 mm,-0.7±2.7 mm/-0.9±4.5 mm/-0.8±4.1 mm,-0.8± 2.6 mm/-0.3±4.4 mm/-1.1±4.0 mm, and-0.6±2.9 mm/-0.7±5.1 mm/-0.9±3.9 mm, respectively. There were no significant differences between the four registration methods.The margins for extension in the left-right, anterior-posterior, and cranial-caudal directions were calculated as 4.7 mm, 5.2 mm, and 6.5 mm, respectively.Conclusions With a comprehensive consideration of various factors, a default setting of automatic registration and manual fine adjustment is appropriate for CBCT-guided radiotherapy for prostate cancer.The margins for extension in the left-right, anterior-posterior, and cranial-caudal directions are 4.7 mm, 5.2 mm, and 6.5 mm, respectively.

Objective To observe the motions of the rectum and bladder by image?guided radiotherapy ( IGRT) and to analyze their impact on treatment. Methods Eighteen patients with prostate cancer undergoing intensity?modulated radiotherapy ( IMRT) were enrolled in the study and 247 cone?beam computed tomography ( CBCT) images were obtained from this study. The clinical target volume, bladder, and rectum were contoured on all simulated CT and CBCT to examine their volume and position changes. The dose distributions were recalculated based on the data of the x?, y?, and z?axis setup errors. The doses to planning target volume ( PTV) and organs at risk were calculated in the replanning, and their impact on treatment was analyzed. Comparison of the planning and replanning results was made by paired t?test. The effects of displacements and volumes of the bladder and rectum on target doses were analyzed by Pearson correlation method. Results Great changes in the volumes of the bladder and rectum were observed during the treatment. For the planning and replanning results, PTVD95% was 7777. 37 cGy vs. 7628. 56 cGy ( P=0. 027), PTV Dmin was 87. 91 cGy vs. 83. 35 cGy (P=0. 000), and RVP was 5. 89% vs. 8. 31%(P=0. 000). There were correlations between PTVD95% and the motions of the bladder and rectum, with correlation coefficients of 0. 296 and 0. 177, respectively. The correlation coefficient between rectal volume and PTVD95% was 0. 115, indicating a certain correlation. There is a certain correlation between and PTV Dmin and bladder volume, with a correlation coefficient of?0. 128. Conclusions The recovery of the state during localization for the bladder and rectum, especially the latter, has great significance to ensure the target dose and reduce exposure of the rectum in the IMRT for prostate cancer.

OBJECTIVETo build a protocol of separation and induction of megakaryocytes derived from cord blood mononuclear cells.

METHODSRed blood cells were precipitated by hydroxyethyl starch (HES). Mononuclear cells were obtained by density gradient centrifugation with Ficoll. The inducing efficiencies of megakaryocytes by using of different cytokine cocktails and culture media were analyzed.

RESULTSThe best choice for erythrocyte sedimentation and high efficiency of nucleated cells retrieving were obtained by using of 1.5% HES. The isolated cord blood mononuclear cells were cultured with domestic serum-free medium supplemented with 116t (IL-11, IL-6, TPO), st36(SCF, TPO, IL-3, IL-6), pt36 (PDGF,TPO,IL-3,IL-6) or pst36 for 7 days. St36 group (50 ng/ml SCF, 50 ng/ml TPO, 20 ng/ml IL-3 and 50 ng/ml IL-6) yielded the most CD41/CD61 positive [(6.79±1.97)×10⁴]. The cell viability [(82.85 ± 0.64)%] of st36 group by using of imported serum-free medium was better than [(60.90±6.93)%] that in domestic medium on day 7 after induction, and CD41/CD61 positive cells count [(18.60±1.97)×10⁴] were more than domestic serum-free medium group. Therefore, we chose imported serum-free medium containing st36 to induce cord blood mononuclear cells. After a prolonged culture, the total cell numbers increased accompanied with an elevated percentage of CD41/CD61 positive cells, which reached (54.27 ± 6.31)% on day 14. Wright-Giemsa staining showed that different phase cells, such as megakaryoblast, promegakaryocyte and granular megakaryocyte, occurred after 10 days'culture. Clone forming unit-megakarocytes (CFU-MK) assay showed that the colonies count increased with the prolonged incubation. CFU-MK colonies were [1 236.0±32.9] on day 14, which was higher than that in medium without induction (P<0.01). Platelets from megakaryocytes showed agglutination function after 10 days'culture.

CONCLUSION1.5% HES was the best solution to precipitate erythrocytes. The combination of an imported serum-free medium with IL-3, IL-6, SCF and TPO showed better induction efficiency than domestic medium or other cytokine cocktails. Meanwhile, induced megakaryocytes produced functional platelets.

Cell Culture Techniques ; Cell Differentiation ; Cell Division ; Cell Separation ; methods ; Cells, Cultured ; Culture Media, Serum-Free ; Fetal Blood ; cytology ; Humans ; Megakaryocyte Progenitor Cells ; cytology