Objective To study changes and effects of transforming growth factor-?1(TGF- ?1) and its receptorIII (TGF-?1 R III) in children with idiopathic thrombocytopenic purpura (ITP). Methods Bone marrow were respectively collected from 28 children with acute idiopathic thrombocytopenic pupura(AITP),16 children with chronic idiopathic thrombocytopenic purpura(CITP) and 20 comparably normal children; Percoll density gradient and immunomagnetic beads methods were used to purify megakaryocytes from bone marrow; ABC- ELISA was used to detect TGF - ?1 in bone marrow; in situ hybridization was used to detect TGF-?1 RIIImR-NA expression on megakaryocytes.Results In AITP and CITP group, the levels of TGF-?1 and TGF-?1 RIIImRNA were significant higher than those in control group(P

Objective: To investigate the mechanism of scorpion venom active peptides(SVAPs) on platelet aggregation. Methods: Platelet aggregation in vivo and vitro was determined by turbidimetry. Carotid thrombosis model was induced by electrostimulation. The determination of 6 keto PGF 1 and TXB 2 were performed by radioimmunoassay. Results: SVAPs 0.125,0.25,0.5mg?ml -1 significantly inhibited the rabbit platelet aggregation triggered by thrombase 0.03u.ml -1 , ADP 10u.ml -1 in vitro( P

Objective To observe the protective effect of acetylcysteine against radiation pneumonia.MethodsTotal of 80 patients who were inoperable were randomly allocated into treatment group and control group.Using conformal radiation technology and the total dose was 65 ～ 75Gy.The patients in treatment group were given acetylcysteine and radiotherapy;the patients in control group were given radiotherapy only.ResultsAll patients were treated radiotherapy.The effective rate( CR and PR) of treatment group was 90％,and that of control group was 85％(P ＞ 0.05);The incidence of acute radiation pneumonia and pulmonary fibrosis in treatment group were 15％ and 20％,respectively;and that of control group were 33％ and 45％ respectively.There was significant difference between the two groups ( P ＜ 0.05 ).ConclusionUsing acetylcysteine during radiotherapy could prevent radiation pneumonia in the non-small cell lung cancer patients.

Purpose To investigate the dosimetric differences among RapidArc (RA) plans which were designed on different target volumes in hepatocellular carcinoma (HCC).Methods A total of 10 HCC patients underwent 3D-CT scan under free breathing ( FB),end inspiration hold ( EIH ) associated with active breath coordinator (ABC) and 4D-CT scan.The 4D-CT were sorted into 10 sets of CT images according to respiratory cycle.The gross tumor volume (GTV) was manually contoured on different CT images.The individual internal gross target volume ( IGTV1 ) was obtained from 4D-CT,and the individual margins from GTVFB to IGTV1.IGTV2 were obtained from GTVFB using individual margins.The planned target volumes (PTV-1,PTV-2,PTV-3 and PTV-4 ) were obtained from GTVFB,IGTV1,IGTV2 and GTVEIA applying different margins.The RA plans (RA1,RA2,RA3 and RA4 ) were designed from different PTVs,and for RA1,RA2 and RA3 the simple 358° arc were used,while three 135° arcs were used for RA4.The dosimetric differences were compared.Results The PTV-1 and PTV-3 were larger than PTV-2 and PTV-4; the mean values of PtV-1/PTV-2 and PTV-1/PTV-4 were 2.5 and 1.9,respectively.There were no significant differences in conformal index,homogeneity index,maximum dose,and minimum dose of PTV among 4 RA plans.The irradiation dose of normal liver of RA3 and RA4 were 8.23 Gy and 7.63 Gy respectively,both significantly lower than those of RA1 and RA2 (10.21 Gy,9.62 Gy,x2 =10.68,P ＜0.05 ),and the V30of RA3 and RA4 were 5.24％ and 5.05％ respectively,both significantly lower than those of RA1 and RA2 (7.76％,6.12％,x2 =14.76,P ＜ 0.05 ).There were no significant differences in irradiation doses of stomach and duodenum among different plans.Conclusions Using 4D-CT or ABC technology with RapidArc in HCC can define the target volume accurately and achieve prefect dose distribution sparing more normal liver volume,compared to the traditional margins.4D-CT and ABC play similar roles in sparing normal liver.

We constructed different N-terminal truncated variants based on Bacillus acidopullulyticus pullulanase 3D structure (PDB code 2WAN), and studied the effects of truncated mutation on soluble expression, enzymatic properties, and application in saccharification. Upon expression, the variants of X45 domain deletion existed as inclusion bodies, whereas deletion of CBM41 domain had an effective effect on soluble expression level. The variants that lack of CBM41 (M1), lack of X25 (M3), and lack both of CBM41 and X25 (M5) had the same optimal pH (5.0) and optimal temperature (60 degrees C) with the wild-type pullulanase (WT). The K(m) of M1 and M5 were 1.42 mg/mL and 1.85 mg/mL, respectively, 2.4- and 3.1-fold higher than that of the WT. k(cat)/K(m) value of M5 was 40% lower than that of the WT. Substrate specificity results show that the enzymes exhibited greater activity with the low-molecular-weight dextrin than with high-molecular-weight soluble starch. When pullulanases were added to the saccharification reaction system, the dextrose equivalent of the WT, M1, M3, and M5 were 93.6%, 94.7%, 94.5%, and93.1%, respectively. These results indicate that the deletion of CBM41 domain and/or X25 domain did not affect the practical application in starch saccharification process. Furthermore, low-molecular-weight variants facilitate the heterologous expression. Truncated variants may be more suitable for industrial production than the WT.
Bacillus ; enzymology ; Glycoside Hydrolases ; metabolism ; Molecular Weight ; Protein Conformation ; Sequence Deletion ; Substrate Specificity ; Temperature

Objective:To investigate the application effects of non-invasive ultrasound cardiac output monitoring (USCOM) in fluid resuscitation guidance and hemodynamic evaluation of patients with sepsis.Methods:A total of 80 patients with sepsis who were treated in The Second Hospital of Jiaxing from January 2021 to December 2022 were included in this single-blind randomized controlled study. These patients were randomly divided into a control group ( n = 40) and an observation group ( n = 40). In the control group, continuous cardiac output indicated by pulse waveform monitoring was used to guide fluid resuscitation and monitor hemodynamic status, while in the observation group, USCOM was used to guide fluid resuscitation and monitor hemodynamic status. The fluid intake and outflow at 24, 48, and 72 hours after admission to the ICU were compared between the two groups. The changes in arterial blood lactate and oxygenation index at 1, 2, 3, 5, and 7 days after admission to the ICU were compared between the two groups. The time of admission to ICU, the length of ICU stay, and changes in hemodynamic indicators were compared between the two groups. The incidence of death within 28 days after admission to the ICU was compared between the two groups. Results:The liquid intake and output in the observation group at 24, 48 , and 72 hours after admission to the ICU were (4 178.13 ± 327.19) mL, (7 763.63 ± 324.08) mL, and (10 501.38 ± 376.74) mL, respectively, which were significantly lower than (4 527.35 ± 276.84) mL, (8 778.15 ± 361.42) mL, and (11 589.12 ± 413.27) mL in the control group ( t = 5.15, 13.22, 12.30, all P < 0.001). The arterial blood lactate levels in the observation group were significantly lower than those in the control group at 1, 2, 3, 5, and 7 days ( t = 5.73, 6.73, 9.98, 12.25, 14.47, all P < 0.001). There was no significant difference in oxygenation index between the two groups on the 1 st day ( P > 0.05). The oxygenation index in the observation group at 2, 3, 5 and 7 days was significantly higher than that in the control group ( t = -4.31, -8.19, -5.28, -9.44, all P < 0.05). The time of admission to ICU and the length of ICU stay in the observation group were (10.15 ± 2.43) days and (16.51 ± 1.36) days, respectively, which were significantly shorter than (12.75 ± 2.87) days and (17.68 ± 1.59) days in the control group ( t = 4.37, 3.54, both P < 0.05). After 24 hours of resuscitation, cardiac output, stroke output, and cardiac index in the observation group were (5.89 ± 0.51) L/min, (57.71 ± 3.82) mL, and (3.31 ± 0.35) L·min -1·m -2, respectively, which were significantly higher than (5.30 ± 0.37) L/min, (50.06 ± 3.25) mL, and (2.85 ± 0.34) L·min -1·m -2 in the control group ( t = -5.92, -9.65, -5.96, all P < 0.001). There was no significant difference in the 28-day mortality rate between the two groups ( P > 0.05). Conclusion:The guidance of USCOM on fluid resuscitation and hemodynamic status assessment in sepsis patients has an obvious effect, which can promote the improvement of patient oxygenation index, and shorten the time of admission to the ICU and the length of hospital stay.

OBJECTIVETo study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A.

METHODSEighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment.

RESULTSIn 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment.

CONCLUSIONSDDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.

Child ; Child, Preschool ; Deamino Arginine Vasopressin ; therapeutic use ; Factor VIII ; analysis ; Hemophilia A ; blood ; drug therapy ; Humans ; Infant ; Male ; Partial Thromboplastin Time

Neomycin-resistance gene is widely used as a selectable marker in eukaryotic expression vector. It codes neomycin phosphotransferase II (NPT II) which confers resistance to various aminoglycoside antibiotic such as G418 and kanamycine. In this work, by site-directed mutagenesis the neo gene mutant was obtained. The expression vector pmDNA using the neo gene mutant as selectable marker has been constructed. After inserting interest luciferase gene, the expression plasmid pmDNAluc + was stably transfected CHO-K1 cells. As a result, the expression positive ratio reaches to approximate 95% and the ratio of high expression colonies is apparently higher than the controls.
Amino Acid Sequence ; Base Sequence ; Drug Resistance ; genetics ; Genetic Markers ; Genetic Vectors ; Kanamycin Kinase ; genetics ; Molecular Sequence Data ; Mutation

OBJECTIVETo investigate the influence of gene transduction mediated by lentivirus vector on human CD34+ cord blood cell (CBCs) gene expression.

METHODSCD34+ cells were isolated and transduced with the third-generation self-inactivating ( SIN) lentiviral vector carrying green fluorescent protein (GFP). The total RNA from transduced cells was extracted and the differences of genotypes between the transduced and non-transduced CD34+ cells were determined with cDNA microarray analysis.

RESULTSIn 23000 genes two were upregulated and six downregulated. These changes were not confirmed by semi-quantitative RT-PCR method.

CONCLUSIONSLentiviral vector used in this study do not influence significantly on the gene expression of CD34+ CBCs, and the vector system may be a useful and safe one in clinical gene therapy.

Antigens, CD34 ; Cells, Cultured ; Fetal Blood ; cytology ; Gene Expression Profiling ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Oligonucleotide Array Sequence Analysis ; Transduction, Genetic ; Transfection

BACKGROUNDPrevention is presently the only available method to limit radiation-induced lung morbidity. A good predictor is the key point of prevention. This study aimed to investigate if [(18)F]2-fluoro-2-deoxyglucose (FDG) uptake changes in the lung after radiotherapy could be used as a new predictor for acute radiation pneumonitis (RP).

METHODSForty-one patients with lung cancer underwent FDG positron emission tomography/computed tomography (FDG-PET/CT) imaging before and after radiotherapy. The mean standardized uptake value (SUV) was measured for the isodose regions of 0 - 9 Gy, 10 - 19 Gy, 20 - 29 Gy, 30 - 39 Gy, 40 - 49 Gy. The mean SUV of these regions after radiotherapy was compared with baseline. The mean SUV in patients who developed RP was also compared with that in those who did not. The statistical difference was determined by matched pair t test. The Radiation Therapy Oncology Group (RTOG) criteria were used for diagnosis and grading of RP.

RESULTSWith a median follow-up of 12 months, 11 (26.8%) of the 41 patients developed grade 2 and above acute RP. The mean SUV of regions (10 - 19 Gy, 20 - 29 Gy, 30 - 39 Gy, 40 - 49 Gy) increased after radiation therapy in all 41 patients. The mean SUVs after radiation therapy were 0.54, 0.68, 1.31, 1.74 and 2.27 for 0 - 9 Gy, 10 - 19 Gy, 20 - 29 Gy, 30 - 39 Gy and 40 - 49 Gy, respectively. Before the radiation therapy, the mean SUV in each region was 0.53, 0.52, 0.52, 0.53 and 0.54, respectively. These patients had significantly higher FDG activities in regions receiving 10 Gy or more (P < 0.001). Compared with their counterparts, the elevation of SUV was significantly greater in those patients who developed acute RP subsequently.

CONCLUSIONThe mean SUV of the lung tissue may be a useful predictor for the acute RP. FDG-PET/CT may play a new role in the study of the radiation damage of the lung.

Aged ; Aged, 80 and over ; Female ; Fluorodeoxyglucose F18 ; Humans ; Male ; Middle Aged ; Positron-Emission Tomography ; methods ; Radiation Pneumonitis ; diagnosis ; etiology ; Tomography, X-Ray Computed ; methods