Objective To study the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSCs) on the resistance of K562cell atd mechanism in vitro.Method MSCs were obtained from AL children bone marrow after derivation, cultivation and identification.The coculture of MSCs and K562 and K562 suspension were established.Effects of MSCs on the growth of K562 cells were investigated in vivo.The two kinds of cells treated with different concentration of adriamycin (ADM) and the rate of apoptosis was evaluated by flow cytometry.Cell cycle was determined by flow cytometry.RT-PCR was used to detect Bcl-2 and Bax in K562 cells.Result Compared with the cell growth curve of K562 alone, the K562 cell co-cultured with MSCs grew slower and the exponential phase of growth was not obvious.The apoptosis index of the K562 cells co- clutured with MSCs was (9.19 ±0.53)% examined by flow cytometry, and that of the K562 cells alone was 4.00 ± 0.37% respectively( P ＜ 0.05 ).The percentage of cells at G0/G1 phase was (50.2 ± 2.26) % and that at S phase was (37.03 ± 3.50) % in the group of K562 alone, but those of the K562 cells co - cultured with MSCs were (80.95 ± 3.83) % and ( 17.40 ± 1.50)% respectively( P ＜0.05).The result of RT-PCR suggested expression of Bcl-2/Bax of the K562 cell co-cultured with MSCs was higher than K562 alone.Conclusion ALL children MSCs suppressed the growth of K562 cell in vitro.Adhesion made K562 depress sensitive to ADM.The mechanism was perhaps caused by adhesion with MSCs, K562 cell cycle was changed and related to Bcl-2 gene high level expression.

OBJECTIVEThe automatic generation of planning targets and auxiliary contours have achieved in Eclipse TPS 11.0.

METHODSThe scripting language autohotkey was used to develop a software for automatically generated contours in Eclipse TPS. This software is named Contour Auto Margin (CAM), which is composed of operational functions of contours, script generated visualization and script file operations. RESULTS Ten cases in different cancers have separately selected, in Eclipse TPS 11.0 scripts generated by the software could not only automatically generate contours but also do contour post-processing. For different cancers, there was no difference between automatically generated contours and manually created contours.

CONCLUSIONThe CAM is a user-friendly and powerful software, and can automatically generated contours fast in Eclipse TPS 11.0. With the help of CAM, it greatly save plan preparation time and improve working efficiency of radiation therapy physicists.

Objective To investigate the expression and significance of glucogen synthase kinase-3β (GSK-3β) in the pathogenesis of chronic allograft nephropathy (CAN) in rats.Methods Kidneys of Fisher (F344) rats as donors were orthotopically transplanted into Lewis (LEW) rats as recipients.The renal function and histopathological changes were observed at 4,8,12,16,and 24week post-transplantation.Phosphorylated GSK-3β (p-GSK-3β) protein and mRNA expression was determined by using immunohistological assays and RT-PCR respectively.Results Our data showed that 24-h urinary protein excretion in CAN rats was increased significantly at week 16 as compared with F344/LEW controls.Allografts showed markedly increased mononuclear cells infiltration and presented with severe interstitial fibrosis and tubular atrophy at 16 and 24 week post-transplantation.p-GSK-3β expression (protein/mRNA) was down-regulated in rat kidneys with CAN,and the decrease became more significant over time after transplantation.p-GSK-3β expression was correlated significantly with 24-h urinary protein excretion,serum creatinine levels,tubulointerstitial mononuclear cells infiltration,smooth muscle cells migration in vascular wall,and interstitial fibrosis.Conclusion It was concluded that GSK-3β down-regulation was the key event that may be involved in mononuclear cells infiltration and vascular SMCs migration at early stage,and interstitial fibrosis and allograft nephroangiosclerosis at later stage of CAN pathogenesis in rats.

Objective To investigate the role and mechanism of phosphate myosin light chain (pMLC) in the rat kidney of chronic allograft nephropathy (CAN) model. Methods The left donor kidneys from Fisher (F344) rats were orthotopically transplanted into Lewis recipients. Meanwhile, the F344 rats and LEW rats with resection of the right kidney served as control groups. Animals were harvested respectively at the 4th, 8th and 12th week after transplantation. The creatinine clearance rate (CCr) was calculated by urine creatinine of 24-h urine. Blood samples were collected from rats for determination of serum creatinine. The expression of pMLC was detected by using Western blotting and immunohistochernistry, and that of integrin-linked kinase (ILK) by using immunohistochemistry. Results Mononuclear cells infiltration of allografts was markedly aggravated as compared to the controls. Allografts got severe interstitial fibrosis and tubular atrophy at 12th week after transplantation. The expression of pMILC and ILK was up-regulated in the kidney of CAN rats after transplantation, and increased more significantly as the time went on. The expression of pMILC was significantly correlated with 24-h urine protein excretion (r= 0. 273, P＜0. 05), serum creatinine levels (r = 0. 434, P＜0. 01 ), the number of tubulointerstitial infiltrated mononuclear cells (r = 0. 525, P＜0. 01 ), the number of smooth muscle cells (SMC) in vascular wall (r= 0. 676, P＜0. 01 ) and the extent of interstitial fibrosis (r= 0. 570, P＜0. 01 ).There was a significantly positive correlation between ILK and pMLC in CAN rats at the 4th week after transplantation (r= 0. 778, P＜0. 01 ). Conclusion pMLC might play an key role in CAN, and the over-expression of ILK might be involve in the pathogenesis of CAN.

Aim To establish a LC-MS/MS method for the determination of hydroxysafflor yellow A ( QA ) in human plasma. Methods After being added into 0. 2M ammonium acetate (1∶1,V/V), QA was extrac-ted using solid-phase extraction technique, and the eluent was directly injected into LC-MS/MS systems. Agilent ZORBAX SB C18 (3. 0 × 100 mm, 3. 5 μm) column and isocratic elution system composing of meth-anol and 0. 2 mM ammonium acetate (70 ∶ 30, V/V) provided chromatographic separation of QA and internal standard isorhamnetin-3-O-neohespeidoside ( SLS) fol-lowed by detection with mass spectrometry. The mass transition ion-pair was followed as m/z 611 . 131→490. 900 for QA and m/z 623. 032→298. 800 for SLS. Results The retention time of QA and SLS was 2. 7 min and 3. 9 min respectively, with no interference in human blank plasma. The proposed method showed good linearity over the concentration range of 8. 57 ~4185 μg·L-1 for QA with a correlation coefficient≥ 0 . 9949 . The lower limit of quantitation was 8. 570 μg ·L-1 . The intra-batch and inter-batch precision and accuracy were within 7%. The average matrix effect ranged from 115. 72% to 119. 06% with RSD less than 5%. The average extraction recovery ranged from 77. 75% to 80. 76% with RSD less than 5%. Stability of human samples after 4 h at room temperature, after the three freeze-thaw cycles and after 31 days at -70℃, and post-preparative stability of the processed sam-ples after 24 h was acceptable. Plasma samples with the concentration beyond the upper quantitation limit could be accurately determined after being diluted using 6. 25 times ( V/V ) of human blank plasma. Conclusion Our current LC-MS/MS method is sensitive, accurate and convenient, and is proved to be suitable for the sys-tematic study on clinical pharmacokinetics of QA.

Aim To investigate the influence of sarpog-relate hydrochloride (SH)on the pharmacokinetic pro-file of dextromethorphan (DM),the typical substrate of CYP2D1 /2,in rats when they were administered co-instantaneously.Methods A total of 1 2 SD rats were randomly divided into two groups:the control group (DM,1 0 mg·kg-1 )and the sarpogrelate group (SH, 1 0 mg·kg-1 ;DM,1 0 mg·kg-1 ),which received in-tragastric administration.Plasma samples were collected immediately before and at different time points after drug administration.A LC-MS /MS method was used to determine the concentrations of DM in rat plasma. Pharmacokinetic parameters were analyzed using Drug and Statistics (DAS 2.0).Results There were signif-icant differences in the pharmacokinetic parameters of DM,including T1 2 (2.49 h ±0.93 h vs 1 .47 h ±0.20 h,P <0.05 ),Cmax (325.7 μg·L -1 ±1 33.2 μg· L -1 vs 1 04.5μg·L -1 ±52.4 μg·L -1 ,P <0.05), AUC0 -t(785.5 μg·L -1 ·h ±451 .9 μg·L -1 ·h vs 244.8 μg·L -1 ·h ±1 68.3μg·L -1 ·h,P <0.05) and AUC0 -∞(804.7 μg·L -1 ·h ±445.6 μg·L -1 ·h vs 251 .4 μg·L -1 ·h ±1 73.4 μg·L -1 ·h,P<0.05 )between the two groups.Conclusion SH could significantly inhibit the elimination of DM,the substrate of CYP2D1 /2 in rats.

Community-acquired pneumonia (CAP) is a leading cause of death in children under 5 years old.Bacterial pneumonia is still the primary cause of severe pneumonia.Prevalence of Staphylococcus aureus CAP and its clinical features remain incompletely understood, complicating the empirical selection of antibiotics. Staphylococcus aureus is the main bacterial pathogen of CAP besides Streptococcus pneumoniae.Due to the virulence its bacterial factors involved in nasal colonization and its relationship with the host immune response, staphylococcus aureus is easy to cause lung damage after the virus infection. Staphylococcus aureus also produces panton-valentine leukocidin, and other toxin factors, which lead to the death of host cells after infection, activate humoral immunity, produce a large number of inflammatory factors, further cause primary and secondary acute lung injury, and may cause systemic collapse infection.At pre-sent, there is little in-death research on the relationship between the toxicity of Staphylococcus aureus and the host immune response.The prevalence of methicillin resistant Staphylococcus aureus makes the anti-infection treatment increa-singly difficult.It is urgent to improve the comprehensive diagnosis and treatment measures, improve the treatment rate, reduce the disability rate, and develop therapies targeted at both pathogen virulence factors and host immunomodulation.

Objective:To verify the clinical applicability of " eCura system" , a scoring system for assessing the risk of lymph node metastasis after non-curative endoscopic submucosal dissection (ESD) for early gastric cancer (EGC).Methods:A retrospective analysis was performed on clinicopathological data of 155 patients with EGC, who underwent non-curative ESD confirmed by postoperative pathology in Drum Tower Hospital Affiliated to Medical School of Nanjing University from January 2012 to March 2018. According to the eCura scoring system, the 155 patients were divided to three groups: 100 cases in the low-risk group (0 to 1 point), 46 cases in the intermediate-risk group (2 to 4 points), and 9 cases in the high-risk group (5 to 7 points). Lymph node metastasis rates and prognosis of the three groups were observed and compared.Results:The follow-up time of the 155 patients was 25±15.0 months, of which median follow-up time was 25 months in the low-risk group, 23 months in the intermediate-risk group, and 34 months in the high-risk group. A total of 57 patients underwent additional surgery in the low-risk group, including 3 cases [5.26% (3/57)] of lymph node metastases; 29 patients underwent additional surgery in the intermediate-risk group, including 2 cases [6.90% (2/29)] of lymph node metastases; all 9 patients in the high-risk group underwent additional surgery and 4 cases had lymph node metastasis. Multivariate Logistic regression analysis showed that the risk of lymph node metastasis in the high-risk group was significantly higher than that in the low-risk group ( P=0.003, OR=14.499, 95% CI: 2.513-97.214), while the risk of lymph node metastasis in the intermediate-risk group was slightly higher than that in the low-risk group ( P=0.767, OR=1.326, 95% CI: 0.165-8.594). During follow-up, there was no metastasis or cancer-specific mortality in the low-risk group, and recurrence was found in 3 cases [6.98% (3/43)] of the 43 patients without additional surgery. Among the 17 patients in the intermediate-risk group, who did not undergo additional surgery, 1 case [5.88% (1/17)] had recurrence and 2 cases [11.76% (2/17)] had metastasis including 1 case [5.88% (1/17)] died of brain metastasis. There was no recurrence, metastasis or cancer-specific mortality in 29 patients in the intermediate-risk group, who underwent additional surgery during follow-up. And there was no recurrence, metastasis or cancer-specific mortality in the all 9 patients in the high-risk group received additional surgery after ESD during follow-up. Conclusion:The eCura scoring system could contribute to predict the lymph node metastasis risk in patients after non-curative ESD for EGC. The benefits of additional surgery are limited for low-risk patients, while for intermediate-risk and high-risk patients, additional surgery can effectively improve prognosis.

Background::Previous studies have shown that bufalin exerts antitumor effects through various mechanisms. This study aimed to determine the antineoplastic mechanism of bufalin, an extract of traditional Chinese medicine toad venom, in ovarian cancer.Methods::The 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT), 5-ethynyl-2 ′-deoxyuridine (EdU), and colony formation assays were used to investigate the antiproliferative effect of bufalin on the ovarian cancer cell line SK-OV-3. Molecular docking was used to investigate the combination of bufalin and epidermal growth factor receptor (EGFR) protein. Western blotting was performed to detect the expression of EGFR protein and its downstream targets. Results::Bufalin inhibited the proliferation of SK-OV-3 cells in a dose- and time-dependent manner. Bufalin was confirmed to combine with EGFR protein using molecular docking and downregulate expression of EGFR. Bufalin inhibited phosphorylation of EGFR, protein kinase B (AKT), and extracellular signal-regulated kinase (ERK).Conclusion::Bufalin suppresses the proliferation of ovarian cancer cells through the EGFR/AKT/ERK signaling pathway.

Objective To survey the basic situation of diagnosis and treatment in clinical nuclear medicine across Guangdong province,evaluate the risks of exposure to the radiation workers and public and explore the countermeasures to control radiation exposre.Methods A survey team was set up to survey,by filling questionnaires,the basic information on nuclear medicine practices for workers,equipment,radionuclide,frequency and dose to workers and radiation protection measures.Results A total of 71 nuclear medicine institutions was involved in the survey with 733 radiation workers in 2016 in Guangdong.The average annual effective dose to nuclear medicine workers was (0.55±0.66) mSv per year.The total pieces of nuclear medical equipment was up to 189 in 2016 including 59 SPECT/CT scanners (5 SPECT),28 PET/CT scanners and 54 thyroid scanners.Total activity of 1.15× 10s MBq in radiopharmaceuticals was used in 325 903 examinations and treatments with the number of frequency of 2.97 examinations per 1 000 population.Concluions There have been a rapid progress in practice of nuclear medicine over the past 20 years in Guangdong province with departments of nuclear medicine set up in 18 of 21 cities.Compared with 1998,the activity in radiopharmaceuticals used has increased by 414％ in 2016 and the number of frequency of examination and treatment has increased by 111％.The rapid expansion of nuclear medicine practice has also caused potential risks of radiation to the public and environment.