Objective:To evaluate the cumulative dose of the target volume and organs at risk (OARs) in intensity-modulated radiation therapy (IMRT) for large volume non-small cell lung cancer (NSCLC) based on rigid and deformation registration methods. The dosimetric changes between the initial and second treatment plans were compared.Methods:Thirty patients treated with IMRT for large volume NSCLC with twice 4DCT scans acquired before radiotherapy and after 20 fractions of radiotherapy were recruited. The initial treatment plan (Plan 1) based on the average density projection CT (CT 1-avg) of the first 4DCT images and the second treatment plan (Plan 2) based on the average density projection CT (CT 2-avg) of the second 4DCT images were calculated. Then, the dose distributions of Plan 1 and Plan 2 were accumulated based on rigid and deformation registration methods to obtain Planrig and Plandef, respectively. Finally, the volume changes of gross tumor volume (GTV) and OARs between two CT scans were compared. The dose-volume parameters between Plan 1 and other plans (including Plan 2, Planrig and Plandef) were also statistically compared. Results:Compared with the initial CT scan, the mean volume of GTV and heart on the second CT was decreased by 44.2% and 5.5%, respectively, while the mean volume of ipsilateral lung, contralateral lung and total lung was increased by 5.2%, 6.2% and 5.8%, respectively (all P<0.05). Compared with Plan 1, the D 95%, D 98% and V 100% of target volume IGTV (GTV fusion of 10 4DCT phases) and PTV in Plan 2 did not significantly change (all P>0.05), and those in Planrig and Plandef were decreased (all P<0.05). The dose-volume parameters of spinal-cord, heart, ipsilateral lung and total lung in Plan 2, Planrig and Plandef were significantly lower than those in Plan 1(all P<0.05). Among them, the V 30Gy and D mean of heart were decreased by 27.3%, 16.5%, 15.3% and 15.2%, 6.6%, 5.6%, respectively. The V 20Gy and D mean of total lung were decreased by 15.6%, 4.5%, 3.7% and 15.7%, 6.2%, 5.1%, respectively. Some dose-volume parameters (including D 95% and D 98% of target volume, V 40Gy of heart, V 20Gy and D mean of the ipsilateral lung and the total lung) of Plandef were higher than those in Planrig (all P<0.05). The Dice similarity coefficients (DSCs) of OARs after deformation registration were significantly higher than those after rigid registration ( P<0.05). Conclusions:The dose-volume parameters of OARs significantly differ between Plan 1 and Plan 2. Hence, all these parameters have a large degree of deviation in predicting radiation-induced injury of OARs. Nevertheless, the dose-volume parameters obtained by deformation registration can enhance the prediction accuracy.

Objective:To investigate the clinical features, imaging findings and prognosis of children with overlapping syndrome of myelin oligodendrocyte glycoprotein (MOG) antibody disease and anti-N-methyl-D aspartate receptor (NMDAR) encephalitis (MNOS).Methods:The clinical manifestations, immunological antibodies in blood and cerebrospinal fluid, cranial image, treatment and follow-up of 11 patients diagnosed as MNOS in the Department of Neurology, Beijing Children′s Hospital from January 2011 to April 2019 were analyzed retrospectively.Results:A total of 11 patients, including 4 males and 7 females were analyzed, the age of onset was (10.4±2.3) years. A total of 29 episodes occurred in 11 children. At the last follow-up, 8 cases showed relapsed remission course, the interval of recurrence was 3 to 60 months. The onset symptoms of 11 patients included convulsions (10 cases), lethargy (6 cases), psychosis (6 cases). Among 29 episodes, the common symptoms were convulsions (16 episodes), psychosis (13 episodes),and lethargy (10 episodes). According to the diagnostic criteria of anti-NMDAR encephalitis and MOG-antibody disease, 29 episodes were divided into three phenotypes, including anti-NMDAR encephalitis(4 episodes), MOG-antibody diseases (10 episodes) and overlapping types (15 episodes).Twenty-seven times of acute stage cranial magnetic resonance imaging (MRI) were available, common lesions included cortical focus (22 times), subcortical white matter (7 times), brainstem (9 times). All patients were sensitive to first-line immunotherapy. Eight patients had recurrence during glucocorticoid reduction, 6 of them were treated with additional second-line immunosuppressive therapy, including cyclophosphamide (1 case) and mycophenolate mofetil (5 cases). The follow-up time of patients were 5-99 months. At the last follow-up, all patients were in remission, the pediatric cerebral performance category (PCPC) score was 1 (10 cases) and 2 (1 cases).Conclusions:MNOS mainly affects older children. In the period of acute episodes, convulsions and psychosis are common. The cranial MRI showed extensive brain involvement and mainly in the cortex. The recurrence rates of MNOS are relatively high, patients are sensitive to first-line immunotherapy. No significant neurological dysfunction was left in the remission stage.

OBJECTIVETo evaluate the replication and encapsidation of HBV mutants with the truncated C gene.

METHODSThe HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome.

RESULTSThe C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay.

CONCLUSIONThe C-truncated HBV mutants are replication-deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon.

Cell Line, Tumor ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Plasmids ; genetics ; Transfection ; Virus Replication

OBJECTIVETo cooperate with the study of HBV vector, hygromycin-resistant packaging cell line was developed that allows encapsidation of plasmids into HBV particles.

METHODSFree of packaging signal, HBV genome was inserted into plasmid pMEP4, which expresses the HBV structural proteins including core, pol and preS/S proteins. HepG2 cell lines were employed to transfect with the construct. Hygromycin selection was done at a concentration of 150 micrograms/ml in the culture medium. The hygromycin-resistant clones with the best expressions of HBsAg and HBcAg were theoretically considered as packaging cell line and propagated under the same conditions. It was infected with recombinant retrovirus vector and hen selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSHygromycin-resistant HBV packaging cell line was generated, which harbored an HBV mutant whose packaging signal had been deleted. Expressions of HBsAg and HBcAg were detectable. Infected with recombinant retrovirus pRV-CP, the hygromycin-resistant packaging cell line was found to secrete mutant HBV particles and no wild-type HBV was detectable in the culture medium.

CONCLUSIONAfter the packaging signal was deleted and transfected into HepG2 cell lines, the partial HBV genome lost its ability to form wild-type HBV, but conserves cis-action providing structural proteins for the packaging of the replication-defective HBV.

Cell Line ; Drug Resistance, Viral ; Genetic Vectors ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Hygromycin B ; pharmacology ; Mutation ; Plasmids ; Retroviridae ; genetics ; Transfection ; Virus Assembly

OBJECTIVETo evaluate the indication and operative program of ventricular septal defect with pulmonary atresia (VSD-PA).

METHODSFrom June 1984 to March 2005, there were 32 patients with VSD-PA, which underwent 33 operations. Among them, 15 were males and 17 were females. The ages ranged from 6 months to 9 years. There were 9 patients with aortopulmonary collateral arteries. The operations included central aorta-pulmonary shunts 3 cases, one stage complete repair 27 cases and one stage unifocalization with VSD open 2 cases.

RESULTSThere were early 5 death including one death after aorta-pulmonary shunt and 4 deaths after one stage complete repair. The causes of death were severe low cardiac output (2 patients), respiratory function failure (1 patient), multiorgan function failure (1 patient) and severe infect (1 patient). Twenty-one patients were followed up from 3 months to 15.5 years. Heart function (NYHA) was class I or II in 19 cases and class III or IV in 2 cases.

CONCLUSIONSThe operative indication and methods mainly depend on the developing of pulmonary arteries and aortopulmonary collateral arteries. Completely surgical repair of patients with VSD-PA can be achieved with acceptable mortality.

Cardiovascular Surgical Procedures ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Septal Defects, Ventricular ; complications ; surgery ; Humans ; Infant ; Male ; Pulmonary Atresia ; complications ; surgery ; Retrospective Studies ; Time Factors ; Treatment Outcome

OBJECTIVETo summarize the experience of surgical treatment of single atrium.

METHODSFrom August 1984 to August 2004, there were 33 patients with single atrium in our study. Plastic surgery for mitral valves were performed for 30 cases with mitral insufficiency. Plastic surgery for tricuspid valves were performed for 18 cases with tricuspid valve insufficiency. There were 3 cases only with complete absence of atrial septum. There were 14 cases with left superior vena cava. All new atrial septums were made with patches including 24 autologous pericardial patches and 9 terylene patches. Complicate abnormalities were corrected in the same time. Tow suture techniques were used in operations to prevent conductive system block, and plastic surgery for mitral valves were performed until the mitral valves were sufficiency.

RESULTSThere weren't death and conductive system block after operation in the group. One case was low-grade mitral insufficiency and the others weren't mitral insufficiency. Twenty-five cases were followed up from 3 months to 11 years, and they could work and study normally.

CONCLUSIONSSingle atrium should be operated as early as possible. The key of surgery is to prevent conductive system block, to properly correct mitral insufficiency and to drastically correct complicated abnormality. The new atrial septum should be made by patch and an autologous pericardial patch is the first selection.

Adolescent ; Adult ; Cardiac Surgical Procedures ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Septal Defects, Atrial ; surgery ; Humans ; Male ; Middle Aged ; Mitral Valve Insufficiency ; surgery ; Treatment Outcome ; Tricuspid Valve Insufficiency ; surgery

OBJECTIVETo study the effects of inhaled nitric oxide (NO) on pulmonary vascular resistance in patients after total cavopulmonary connection (TCPC).

METHODSFifty-two patients after TCPC were evaluated, of them 24 patients were administered with inhaled nitric oxide in the early postoperative period. The cardiac index (CI) and pulmonary vascular resistance (PVR) were compared before and after inhaled NO.

RESULTSIn experimental group, after inhaled NO, partial pressure of oxygen in artery/fraction of inspired oxygen increased from 161 +/- 17 to 193 +/- 23 (t = 2.75, P < 0.01); CI from (2.86 +/- 0.24) L.min(-1).m(-2) to (3.13 +/- 0.22) L.min(-1).m(-2) (t = 2.25, P < 0.05); PVR decreased from (4.2 +/- 0.5) U/m(2) to (3.8 +/- 1.4) U/m(2) (t = 2.29, P < 0.05); central venous pressure (CVP) from (17.0 +/- 1.8) mm Hg to (15.0 +/- 1.1) mm Hg, decreased 11.7%. Compared with the control group, respirator time decreased from (86 +/- 27) h to (54 +/- 18) h (t = 2.29, P < 0.05); ICU time from (6 +/- 2) d to (4 +/- 2) d (t = 2.32, P < 0.05); But hydrothorax drainage and length of stay had no significant difference.

CONCLUSIONSThough inhaled NO, there is no significant long-term effects in patients after TCPC, but it may play an important role in the management of low cardiac output syndrome and high cava pressure caused by reactive elevated pulmonary vascular resistance in the early postoperative period of TCPC.

Administration, Inhalation ; Adolescent ; Adult ; Arteriovenous Shunt, Surgical ; methods ; Cardiac Output ; drug effects ; Child ; Child, Preschool ; Female ; Humans ; Male ; Nitric Oxide ; administration & dosage ; therapeutic use ; Postoperative Period ; Pulmonary Artery ; physiology ; surgery ; Time Factors ; Vascular Resistance ; drug effects ; Vasodilator Agents ; administration & dosage ; therapeutic use ; Venae Cavae ; surgery

The aim of this study was to investigate the effects of gemcitabine(GEM) on apoptosis and c-myc gene expression of HL-60 cells, and feasibility of using GEM in therapy of leukemia. The HL-60 cells were cultured in vitro. The expressions of the c-myc mRNA and C-MYC protein were detected by RT-PCR and Western-blot respectively. The cell apoptosis was analyzed by TUNEL staining. The results showed that after the HL-60 cells were treated with 1.0 microg/ml GEM for 12, 24, 36 and 48 hours, the expression of c-myc mRNA was inhibited to various degree. This inhibitory effect displayed time-dependent manner and the most optimal effective time was 24 hours. Compared GEM group with Ara-C group and blank control group, there were statistical differences (p<0.05). After the HL-60 cells were treated with 1.0 microg/ml GEM for 24, 48, 72 hours, C-MYC protein significantly decreased, and the expression of C-MYC protein reached to lowest level at 48 hours after treating with GEM, and with inhibition rate of 94.16%. Compared GEM group with Ara-C group and blank control group, the differences were significant (p<0.01). There was significant difference between cells treated with GEM for 24, 48 and 72 hours (p<0.01). After the HL-60 cells were treated with 1.0 microg/ml GEM for 24 hours, the apoptotic cells increased obviously. The positive rate was 83.67% in GEM-treated group. Compared GEM group with Ara-C group (positive rate 10.67%) and untreated group (positive rate 3.00%), the differences had statistical significance (p<0.01). It is concluded that GEM can induce the apoptosis and down-regulate c-myc gene expression significantly in HL-60 cells and it may be used as a new therapeutic drug for leukemia.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; RNA, Messenger ; genetics

Animals ; COS Cells ; Female ; Genetic Therapy ; Interleukin-12 ; genetics ; Liver Neoplasms, Experimental ; immunology ; therapy ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo evaluate the possibility of hepatitis B virus (HBV) as a vector in liver-targeting gene therapy.

METHODSA fragment containing the small envelope gene of HBV was replaced with the reporter gene green fluorescent protein (GFP) to construct the recombinant HBV vector, which was transfected into HepG2 cells with liposome. The expression of GFP was observed with fluorescence microscope. The HBV cccDNA was testified using semi-nest PCR. The viral particles of the recombinant HBV in culture medium were detected by PCR as well as Southern blot.

RESULTSThe HBV vector carrying the interesting gene of GFP could express the functional protein in the transfected hepatocytes. However, the recombinant HBV vector was replication-deficient, which could not be packed and replicated in the hepatocytes to secrete mature recombinant HBV particles carrying the interesting gene of GFP when transfected solely but could when cotransfected with the recombinant and helper construct which lacked part of 5'-proximal HBV RNA packaging signal epsilon.

CONCLUSIONIt is possible that HBV is reconstructed as a liver-targeting vector for gene therapy.

Cell Transformation, Viral ; Cells, Cultured ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; physiology ; Hepatitis B virus ; genetics ; physiology ; Hepatocytes ; cytology ; virology ; Humans ; Liver ; cytology ; virology ; Recombinant Proteins ; genetics ; Transfection ; Virus Replication