BACKGROUND:Little data have been available concerning the mechanism of drag resistance and anti-apoptosis in leukemic cells of leukemia children.The majority of studies focus on normal bone marrow mesenchymal stem cells (MSCs) and established stroma cells,but not interaction of MSCs and leukemic cells in leukemia children.OBJECTIVE:To explore the effect of MSCs in leukemia children on the proliferation and apoptosis of leukemic cell strain K562/AO_2.DESIGN,TIME AND SETTING:In vitro cytology experiment was performed at the laboratory of Department of Pediatrics,First Affiliated Hospital of Guangzhou Medical College from December 2007 to August 2008.MATERIALS:MSCs were provided by 30 leukemia children admitted to First Affiliated Hospital of Guangzhou Medical College,including 22 acute lymphoblastic leukemia and 8 acute myeloblastic leukemia.Written informed content was obtained from all families.K562/AO_2 was provided by Tianjin Institute of Hematopathy.METHODS:MSCs were isolated and cultured by Ficoll density gradient method.They were cultured in two conditions:the co-culture of MSCs and K562/AO_2 and K562/AO_2 suspension alone.In co-culture group,1×10~8 /L K562/AO_2 cells at log phase were added to confluent MSCs,and free floating K562/AO_2 cells were discarded after 24 hours.MAIN OUTCOME MEASURES:Effect of MSCs on the growth of K562/AO_2 cells was observed;effect of addamycln on K562/AO_2 cell apoptosis was detected by AnnexinV-FITC method.Cell cycle was determined by flow cytomtry,mdrl gene of K562/AO_2 cell was detected by Taqman-MGB probe real-time PCR.RESULTS:Compared with K562/AO_2 alone,the K562/AO_2 cell co-cultured with MSCs grew slower and the log phase of growth was not significant;the rate of apoptosis in earlier period was significantly decreased (P ＜ 0.05);co-cultured K562/AO_2 G_0-G_1 phase increased,but S phase decreased.No changes in mdr1 gene in cells were found between two culture conditions (P ＞ 0.05).CONCLUSION:In vitro cytology has demonstrated that leukemia children MSCs induce drug resistance of K562/AO_2 cells by changing K562/AO2 cell cycle through adhesion to avoid pro-apoptotic effect of drugs but not related with mdr1 gene.

Objective To investigate the role of Ephrin A2 in the regeneration and reinnervation of hair cells in the chick cochlea following kanamycin ototoxicity.Methods 66 newly hatched Roman chickens (3 days old) were randomly divided into experimental group and control groups. Experimental chickens (n=48) received intramuscular kanamycin (200 mg/kg:Sigma,St Louis,MO) for 10 consecutive days and were subsequently sacrificed 2 days before the last injection,and 1,3,7,10,15,21,30,and 60 days after the last injection (n=6 per subgroup). Control chickens (n=18) were untreated and sacrificed 3,13 and 43 days after hatching (n=6 per subgroup). Ephrin A2 protein expression in acoustic ganglia was determined by western blot analysis in all chickens after sacrifice. Results Ephrin A2 protein expression was found and the protein level was almost same in acoustical ganglia of all normal chickens. After kanamycin exposure,the Ephrin A2 protein expression level in the cochlea of the experiment chickens from 2 to 7 days after last kanamycin injection was lower than that in control chickens,respectively. Ephrin A2 expression increased obviously at 15 days after kanamycin last injection. By 30 days after the cessation of kanamycin treatment,the level of Ephrin A2 protein approximated to that in normal control group.Conclusion The expression of Ephrin A2 protein in the acoustical ganglia basically synchronizes with the regeneration and the reinnervation of the hair cells in the chicken cochlea following kanamycin damage,indicating that Ephrin A2 may play an important role in this process.

Objective:To evaluate the clinical advantage and disadvantage of anatomical landmark registration(ALR) and surface registration(SR) in computer-assisted endoscopic sinus surgery(CAESS).Method:Twenty-six patients were selected for the CAESS, the preparatory times and mean target registration errors (TRE) were recorded in order to compare the difference between them two, their convenience and their value were also analyzed.Result:CAESSs were successfully used in 26 cases without complications. The average preparation time of SR was(8.5±1.9)minutes, that of ALR was(6.5±1.7)minutes. In the SR group, the TRE of naso-labial angle was(1.43±0.26)mm, that of front end of middle turbinate was(1.92±0.47)mm, that of front end of inferior turbinate was (1.82±0.49)mm, and that of back end of inferior turbinate was (2.03±0.42)mm. Them in ALR group were (1.58±0.35)mm,(2.05±0.37)mm,(1.92±0.31)mm and (2.48±0.64)mm ,respectively.24 cases (92.2%) were not affected or were slightly affected by the navigation system. The value of navigation was affirmative in 22 cases (84.6%), and its value was mainly related to TRE.Conclusion:The accuracy of surface registration was superior to that of anatomical landmark registration, but the anatomical landmark registration was more convenient and need less preparation time. The value of navigation system is its accuracy, convenience and no disturbance to surgery. The navigation system is more valuable in the complex cases than that in the general case.

PURPOSE: Cancer stem cells have recently been thought to be closely related to tumor development and reoccurrence. It may be a promising way to cure malignant glioma by using glioma stem cell-targeted dendritic cells as a tumor vaccine. In this study, we explored whether pulsing dendritic cells with antigens of glioma stem cells was a potent way to induce specific cytotoxic T lymphocytes and anti-tumor immunity. MATERIALS AND METHODS: Cancer stem cells were cultured from glioma cell line U251. Lysate of glioma stem cells was obtained by the repeated freezing and thawing method. Dendritic cells (DCs) were induced and cultured from the murine bone marrow cells, the biological characteristics were detected by electron microscope and flow cytometry. The DC vaccine was obtained by mixing DCs with lysate of glioma stem cells. The DC vaccine was charactirizated through the mixed lymphocyte responses and cell killing experiment in vitro. Level of interferon-gamma (IFN-gamma) in the supernatant was checked by ELISA. RESULTS: After stimulation of lysate of glioma stem cell, expression of surface molecules of DC was up-regulated, including CD80, CD86, CD11C and MHC-II. DCs pulsed with lysate of glioma stem cells were more effective than the control group in stimulating original glioma cells-specific cytotoxic T lymphocytes responses, killing glioma cells and boosting the secretion of IFN-gamma in vitro. CONCLUSION: The results demonstrated DCs loaded with antigens derived from glioma stem cells can effectively stimulate naive T cells to form specific cytotoxic T cells, kill glioma cells cultured in vitro.
Animals ; Antigens, Neoplasm/immunology ; Apoptosis ; Brain Neoplasms/*therapy ; Cancer Vaccines/*therapeutic use ; Cell Line, Tumor ; Cell Proliferation ; Dendritic Cells/*cytology ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Glioma/*therapy ; Humans ; Interferon-gamma/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplastic Stem Cells/*cytology ; T-Lymphocytes, Cytotoxic/immunology

Objective To explore the changes of SOX9 and WT1 expressions in rat Sertoli cells irradiated by EMP ( electromagnetic pulse),S-HPM ( S-band high power microwave) and X-HPM ( Xband high power microwave).Methods Primary Sertoli cells were isolated from 3-week-old Wistar rats and its purity was immunocytochemistrically indentified with WT1.After exposure to 6 × 104 V/m EMP,100 mW/cm2 S-HPM and X-HPM for 4 min respectively,SOX9 and WT1 expressions in Sertoli cells were determined with real-time PCR and Western blot,respectively.Results SOX9 mRNA expression was decreased at 6 and 12 h post-irradiation of three different bands of electromagnetic microwave ( F =15.20and 4.84,P ＜ 0.05 ).SOX9 protein expression was also decreased at 6 and 24 h after irradiation ( F =8.46 and 7.47,P＜0.05).WT1 mRNA expression was decreased at6 and 12 h (F=13.46 and 5.08,P ＜ 0.05 ),but its protein expression was decreased only at 24 h post-irradiation ( F =10.26,P ＜ 0.05 ).Conclusions Three bands of electromagnetic radiation reduce the expressions of SOX9 and WT1 in rat Sertoli cells,which may provide molecular foundation for genital system hazards induced by microwave radiation.

Objective To calculate the electric field intensity and transmembrane voltage of spherical cells and ellipsoi -dal red blood cells ( RBC) in an extremely low frequency electromagnetic field .Through this calculation , we can provide reference to the search for interaction targets and mechanics between the extremely low frequency electromagnetic field and organisms.Methods The Finite Element Method was used in the numerical computation for the spherical cell model and the ellipsoidal RBC model .Results The electric field intensity of the two types of cells on the cellular membrane was both significantly higher than the applied electric field strength , and the values of the induced field strength and transmembrane voltage varied with the direction of the electric field periodically .Conclusion The cell shape and direction of the applied electric field are not the main determinants of the cellular membrane electric field intensity and the transmembrane voltage compared with electromagnetic parameters .The distribution of the electric field intensity and transmembrane voltage are re-lated to the direction of the applied electric field.

Objective: To evaluate the prognostic value of the preoperative Geriatric Nutritional Risk Index (GNRI) in patients with esophageal squamous cell carcinoma after radical resection. Methods: Clinicopathological and laboratory data of 315 elderly patients with esophageal squamous cell carcinoma who were older than 60 years and underwent radical resection in Tianjin Medical University Cancer Institute and Hospital from January 2008 to December 2012 were retrospectively analyzed. The GNRI formula was as follows:1.489×serum albumin (g/L)+41.7×(current body weight/ideal body weight). According to the GNRI, patients were divided into the normal and abnormal GNRI groups. The χ2 test was used to analyze the relationship between the GNRI and the clinicopathological char-acteristics of patients. The Kaplan-Meier method was used to analyze the survival rate, and survival analysis was conducted using the Log-rank test. Multivariate survival analysis was conducted using the Cox proportional risk regression model. Results: There were 259 patients in the normal GNRI group (GNRI>98) and 56 patients in the abnormal GNRI group (GNRI≤98). The GNRI was closely correlated with age, tumor location, tumor diameter, serum albumin level, body mass index (BMI), and lymph node metastasis (all P<0.05). The 5-year survival rates in the normal and abnormal GNRI groups were 41.2% and 27.0%, respectively, with statistical significance (P=0.002). Univariate analysis showed that age, tumor diameter, serum albumin level, BMI, GNRI, platelet-lymphocyte ratio, tumor invasion depth, and lymph node metastasis were risk factors for the prognosis of patients with esophageal squamous cell carcinoma (all P<0.05). Multivariate analysis showed that the preoperative GNRI (hazard ratio=0.687, 95% confidence interval: 0.487-0.968, P=0.032) was an independent factor affecting the prognosis of patients with esophageal squamous cell carcinoma. Subgroup analysis showed that the survival rates in the normal GNRI group were significantly higher than those in the abnormal GNRI group (P=0.036 and 0.010, respectively), regardless of lymph node metastasis. Conclusions: The preoperative GNRI is associated with malignant biological behav-ior in elderly patients with esophageal squamous cell carcinoma and can be used as a useful indicator for predicting survival after radi-cal resection.

Objective:To evaluate the application value of transabdominal wall suspended laparoscopic appendectomy (LA) in patients with acute suppurative appendicitis or concurrent periappendiceal abscess.Methods:The clinical data of 107 patients with acute suppurative appendicitis or concurrent periappendiceal abscess in Hexi University, Zhangye People′s Hospital from September 2019 to December 2022 were retrospectively analyzed. Among them, 53 patients underwent open appendectomy (OA) (OA group), and 54 patients underwent transabdominal wall suspended LA (LA group). The operation time, postoperative pain score, postoperative hospitalization time, recovery time of gastrointestinal function, postoperative complications and hospital cost were compared between two groups.Results:In the LA group, 2 cases were transferred to OA due to heavy abdominal adhesion and unclear anatomy, and 3 cases could not undergo transabdominal wall suspended LA due to perforation of the root of the appendix or gangrene of the appendix. There was no statistical difference in operation time between two groups ( P>0.05); the postoperative hospitalization time, recovery time of gastrointestinal function, postoperative pain score and total incidence of postoperative complications in LA group were significantly lower than those in OA group: (4.92 ± 1.70) d vs. (7.51 ± 3.96) d, (20.64 ± 7.37) h vs. (35.32 ± 10.13) h, (5.62 ± 1.12) scores vs. (6.83 ± 0.93) scores and 5.56% (3/54) vs. 24.53% (13/53), the hospital cost was significantly higher than that in OA group: (8 325.47 ± 856.22) yuan vs. (6 458.64 ± 2 085.93) yuan, and there were statistical differences ( P<0.05). Conclusions:Transabdominal wall suspended LA has the advantages of wide indications, easy operation, minimal trauma, fast recovery, and fewer complications, but with relatively high hospitalization cost.

Objective:To analyze the differences and similarities of pre-treatment and post-treatment lung microbiome of acute respiratory distress syndrome (ARDS) and find out the change rules of the lung microbiome in the progression of ARDS according to different prognosis.Methods:A retrospective study was conducted. Patients with ARDS caused by severe pneumonia admitted to intensive care unit (ICU) of Jiangmen Central Hospital from February 2019 to January 2020 were enrolled as the study subjects. The patients were divided into pre-treatment (ARDS-preT) group (24 cases), post-treatment survival (ARDS-poT-Survival) group (17 cases), and post-treatment death (ARDS-poT-Dead) group (7 cases). ICU patients with mild pulmonary infection and non-ARDS admitted to ICU during the same period were enrolled as control group (25 cases). The similarities and differences of lung microbiome in four groups were analyzed and compared, and the possible pathogenic bacteria (potential risk factors for death) and probiotics (potential survival and protective factors) related to death caused by ARDS were screened.Results:In terms of pathogenic microorganisms, the positive rates of Escherichia coli and Candida albicans in the ARDS-poT-Dead group were significantly higher than those in the ARDS-poT-Survival group [57.1% (4/7) vs. 5.9% (1/17) and 57.1% (4/7) vs. 0% (0/7), both P < 0.05]. In the screening of background bacteria, the decrease of bacteria in the ARDS-preT group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the control group, the reduced bacteria might be pulmonary probiotics (potential protective factor for ARDS). The screening result was Hydrobacter [ARDS-preT group vs. ARDS-poT-Survival group: 62.5% (15/24) vs. 94.1% (16/17); ARDS-poT-Dead group vs. ARDS-poT-Survival group: 14.3% (1/7) vs. 94.1% (16/17); ARDS-poT-Dead vs. control: 14.3% (1/7) vs. 96.0% (24/25), all P < 0.05]. In the screening of background bacteria, the increase of bacteria in the ARDS-poT-Dead group compared with the ARDS-preT group, the ARDS-poT-Dead group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the control group, and the increased bacteria might be potential pulmonary pathogen (potential risk factor for death of ARDS), which belonged to Enterobacteria: Edwardsiella, Enterobacteriaceae, Escherichia, Klebsiella, Kluyvera, Lelliottia, Pantoea, Raoultella. Conclusions:The results revealed the increase of Escherichia coli or Candida albicans in pulmonary pathogenic microorganisms, or the increase of Enterobacteria in background bacteria may be the risk factors for the death of ARDS. Additionally, background bacteria Hydrobacter probably is a protective factor for the survival of ARDS. Whether it can be used as a novel treatment for ARDS is worth further investigation.

Myocardial fibrosis is a key link in the occurrence and development of diabetic cardiomyopathy. Its etiology is complex, and the effect of drugs is not good.Cardiomyocyte apoptosis is an important cause of myocardial fibrosis. The purpose of this study was to investigate the effect of gaseous signal molecule sulfur dioxide (SO2 ) on diabetic myocardial fibrosis and its internal regulatory mechanism. Masson and TUNEL staining, Western-blot, transmission electron microscopy, RT-qPCR, immunofluorescence staining, and flow cytometry were used in the study, and the interstitial collagen deposition, autophagy, apoptosis, and changes in phosphatidylinositol 3-kinase (PI3K)/AKT pathways were evaluated from in vivo and in vitro experiments. The results showed that diabetic myocardial fibrosis was accompanied by cardiomyocyte apoptosis and down-regulation of endogenous SO2 -producing enzyme aspartate aminotransferase (AAT)1/2 . However, exogenous SO2 donors could up-regulate AAT1/2 , reduce apoptosis of cardiomyocytes induced by diabetic rats or high glucose, inhibit phosphorylation of PI3K/AKT protein, up-regulate autophagy, and reduce interstitial collagen deposition. In conclusion, the results of this study suggest that the gaseous signal molecule SO2 can inhibit the PI3K/AKT pathway to promote cytoprotective autophagy and inhibit cardiomyocyte apoptosis to improve myocardial fibrosis in diabetic rats. The results of this study are expected to provide new targets and intervention strategies for the prevention and treatment of diabetic cardiomyopathy.