OBJECTIVETo investigate the capability of semi-quantitative and quantitative parameters of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to predict the response to concurrent chemoradiotherapy( CCRT) in patients with non-small cell lung cancer (NSCLC).

METHODSA total of 24 patients with stage III or IIIB NSCLC, who underwent 3.0T DCE-MRI before CCRT, were enrolled in this study. Semi-quantitative and quantitative parameters were calculated by Funtool and Omnikinetics software. The relationship between these obtained parameters and tumor response was evaluated by Spearmen' s correlation analysis. The patients were classified into two groups according to the tumor regression rate after treatment, as response group (group A) and non-response group ( group B). Mann-Whitney U test was used to compare the parameters of responders and non-responders. The value of the parameters on predicting response was calculated by receiver operating characteristic curve (ROC).

RESULTSThe tumor regression rate after treatment was negatively correlated with time to peak (TTP) and the extravascular-extracellular volume fraction (Ve), and was positively correlated with signal enhancement ratio (SERmax) and volume transfer constant (Ktrans) (P < 0.05 for all). Statistical significant differences were found between group A and group B both in semi-quantitative and quantitative parameters (P < 0.05). Group A had a lower TTP value [(34.66 ± 16.37) s vs. (44.09 ± 17.41) s] and Ve value [(0.19 ± 0.03) vs. (0.25 ± 0.05)] than group B, whereas group A had a higher SERmax [(166.50 ± 44.95)% vs. (113.57 ± 46.62)%] and Ktrans [(0.41 ± 0.17) min(-1) vs. (0.28 ± 0.12) min(-1)] than group B (P < 0.05 for all). The ROC analysis indicated that when setting the threshold of Ve on ≤ 0.21 for predicting response, the specificity, sensitivity and accuracy were 85.7%, 80.0% and 83.3%, respectively, with an area under curve of 0.875 (P < 0.001).

CONCLUSIONSBoth the semi-quantitative and quantitative DCE-MRI parameters are helpful for predicting the response after CCRT of NSCLC. Quantitative parameters seem to be more meaningful than semi-quantitative parameters.

Carcinoma, Non-Small-Cell Lung ; pathology ; therapy ; Chemoradiotherapy ; methods ; Contrast Media ; Humans ; Lung Neoplasms ; pathology ; therapy ; Magnetic Resonance Imaging ; methods ; ROC Curve ; Remission Induction ; Sensitivity and Specificity ; Time Factors

Objective To construct a prokaryotic expression system for major histocompatibility complex class Ⅰ chain-related gene B ( MICB) and to establish an ELISA method for the detection of anti-MICB antibodies in patients with kidney transplantation. Methods The MICB cDNA fragments were ob-tained by RT-PCR with a pair of specific primers. The MICB cDNA and the prokaryotic expression vector pET-28a were digested by two restriction enzymes and ligated to construct the recombinant expression plas-mid pET-28a-MICB. The transformed E. coli BL21 DE3 strains carrying recombinant expression plasmid were induced by IPTG to express MICB protein. The expressed recombinant proteins were identified by Western blot assay and purified by Ni-NTA Spin column. The purified proteins were coupled to ELISA for the detection of anti-MICB antibodies in patients with kidney transplantation. Results Three common MICB fragments contained the exons 2 and 3 were obtained. The recombinant proteins were expressed in E. coli BL21 DE3 strains carrying pET-28a-MICB and successfully purified by the Ni-NTA Spin column. Results of the Western blot assay confirmed that the obtained proteins were the target proteins. The ELISA method was successfully established and used for the detection of anti-MICB antibodies in 24 patients with kidney trans-plantation. The absorbance values indicated that the sensitivities of three recombinant MICB proteins were different. Conclusion The expression system for MICB gene was successfully constructed. The established ELISA for the detection of anti-MICB antibodies would pave the way for further investigation on the correla-tions between MICB protein and transplantation immunity.

Objective: To explore the correlation between depression severity and neurocognitive function in patients with late-onset depression . Methods: The patients with late-onset depression treated in Jinhua Second hospital from February 2015 to December 2017 were assigned into the mild,moderate and severe groups according to the severity of depression assessed by the Hamilton Depression Scale-17（HAMD-17）. At the same time,some healthy persons were selected as the control group. Wisconsin Card Sorting Test（WCST）,Verbal Fluency Test（VFT）and Stroop Test were carried out,and the scores of these tests were compared in the four groups. The correlations of WCST, VFT, Stroop Test and HAMD-17 scores were analyzed . Results: There were 32,28,35 and 35 subjects involved in the mild,moderate,severe and control group,respectively. The subjects of the mild group,moderate group and severe group had more total errors,perseverative responses and perseverative errors than the control group,and less percent conceptual level responses than the control group （all P<0.05）. The total errors,perseverative responses,perseverative errors and percent perseverative errors increased and the percent conceptual level responses decreased gradually with the severity of depression（all P<0.05）. The correct numbers of Stroop-consistent group and VFT in the severe group were less than those in the control,mild and moderate group（all P<0.05）,which was significantly different between the mild,moderate and control group （P>0.05）. The HAMD-17 scores were negatively correlated with the correct numbers of Stroop congruent group（r=-0.448,P<0.001）and VFT（r=-0.401,P<0.001）,and were positively correlated with perseverative responses in the WCST（r=0.784,P<0.001） . Conclusion The neurocognitive impairment in patients with late-onset depression aggravated with the severity of depression.

Objective To propose an optimal therapeutic scheme, finite element analysis was used to evaluate the biome?chanical status of femoral head under daily activities. Methods Non?necrotic femoral head volunteers (11 people) and necrotic femoral head patients (7 cases) were included in this study. All these participants were divided into two groups:control group and abnormal group. The DICOM imaging files of each participant were imported into ScanIP software to form a mask showing the prox?imal femurs. Then the finite element models of proximal femurs were progressively reconstructed. The generated models were im?ported into Abaqus, and the forces were added on these models to simulate daily activities, including standing, sitting, upstairs, and squatting etc. The loads and boundary conditions of digital daily activities simulation were collected from Orthoload database. All of the femoral finite element models were progressed in Abaqus under above activity conditions. After the models were pro?cessed, stress concentrating region was defined in each models respectively, which expressed the main bearing structure in each model based on the result of finite element analysis. At last, the elastic modulus (E?modulus) and strain of the stress concentrating region in each femoral model were extracted and calculated. Results There was no significant difference in age, weight, and height between these two groups. In control group, no significant differences of E?modulus (7 125±745 MPa on left, and 7 112±751 MPa on right) and strain (micro?strain:952 on left, and 954 on right)were found between their bilateral proximal femurs;however in abnormal group, the patients' E?modulus (5 798±1036 MPa on normal side, and 6 667±888 MPa on necrotic side) and strain (mi?cro?strain:1 261 on normal side, and 987 on necrotic side) of necrotic side were significantly lower than normal side. As to all the daily activities which were considered in this study, in abnormal group, the E?modulus among each postures of abnormal side were significantly lower than the E?modules of healthy side;the micro?strain in downstairs was also significantly lower than other pos?tures. Conclusion Daily activities with different intensity could generate different mechanical effects on femoral head, espe?cially on necrotic femoral head. The higher intensity of posture, the greater damage would be found in necrotic femoral head.

OBJECTIVE: To study the clinical effect and safety of different maintenance doses of caffeine citrate in the treatment of apnea in very low birth weight preterm infants. METHODS: A total of 78 very low birth weight preterm infants with primary apnea were enrolled who were admitted from January 2016 to January 2018. They were randomly divided into high-dose caffeine group with 38 children and low-dose caffeine group with 40 children. Both groups received a loading dose of 20 mg/kg caffeine citrate, and 24 hours later, the children in the high-dose caffeine group were given a maintenance dose of 10 mg/kg, and those in the low-dose caffeine group were given a maintenance dose of 5 mg/kg. The two groups were compared in terms of response rate and incidence rate of adverse events. RESULTS: The high-dose caffeine group had a significantly higher response rate than the low-dose caffeine group (71% vs 48%; P<0.05). Compared with the low-dose caffeine group, the high-dose caffeine group had significantly shorter duration of apnea and time of caffeine treatment (P<0.05). There were no significant differences between the two groups in length of hospital stay and incidence rates of tachycardia, feeding intolerance, bronchopulmonary dysplasia, necrotizing enterocolitis, and intracranial hemorrhage (P>0.05). There was no significant difference in the mortality rate between the two groups (P>0.05). CONCLUSIONS Higher maintenance dose of caffeine citrate has a better clinical effect than lower maintenance dose of caffeine citrate in the treatment of apnea in very low birth weight preterm infants, without increasing the incidence rates of adverse drug reactions and serious complications in preterm infants.
Apnea ; drug therapy ; Caffeine ; therapeutic use ; Child ; Citrates ; therapeutic use ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Infant, Very Low Birth Weight ; Prospective Studies

Objective:To identify the differentially expressed proteins that caused hippocampal damage after liver ischemia-reperfusion (I/R) in rats.Methods:Eighteen clean-grade healthy juvenile male Sprague-Dawley rats, aged 2 weeks, weighing 20-30 g, were divided into 2 groups ( n=9 each) using a random number table method: sham operation group (S group) and liver I/R group (IR group). A rat model of liver I/R injury was prepared by restoring perfusion after 1 h of liver ischemia. The rats were sacrificed after being anesthetized at day 3 of reperfusion, and the hippocampal tissue was isolated and analyzed to obtain gene expression profiles. Differentially expressed genes were identified using the R software, and further protein interaction networks were constructed through Cytoscape and Kyoto Encyclopedia Genes and Genomes pathway analysis to determine the differentially expressed proteins. Quantitative real-time polymerase chain reaction and Western blot were used for validation. Results:A total of 45 differentially expressed proteins were identified by the proteomic analysis of hippocampal tissues, including 36 significantly up-regulated proteins and 9 significantly down-regulated proteins. The proteins with significant expression related to injury were identified from the PPI network complex using the CytoHubBA plug-in cystscape: Ras-related C3 botulinum toxin substrate (RAC2), HRAS, phosphatidylinositol-3-kinase inhibitor phosphatase and tensin homologue (PTEN), and N-methyl-D-aspartate ionotropic glutamate receptor 2b (GRIN2b). The results of quantitative real-time polymerase chain reaction and Western blot showed that the expression of RAC2, HRAS, PTEN, and GRIN2b in the hippocampal tissue was significantly up-regulated in IR group compared with S group ( P<0.05). The results of Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that differentially expressed proteins were significantly enriched in the expression of PD-L1 and its checkpoint pathway, long-term potentiation, and regulation of the Wnt signaling pathway in cancer. Conclusions:The mechanism by which liver I/R induces hippocampal injury may be related to the up-regulation of the expression of RAC2, HRAS, PTEN and GRIN2b in rats.

This study was aimed to investigate the effects of different stimulatory factors on proliferation and function of cytokine induced killer (CIK) cells. Peripheral blood mononuclear cells (PBMNC) were separated by Ficoll-Hypacue gradient. According to supplement of different stimulatory factors (CD28 mAb, IL-15 and IL-21), the experiment was divided into five groups:control group (CIK), CB28+IL-15+IL-21 group, IL-15+IL-21 group, CD28+IL-15 group and CD28+IL-21 group. Effects of different stimulatory factors on the proliferation of CIK cells were assayed by an automated hematology analyzer. Changes of granzyme B,perforin and CD107a were detected by flow cytometry. IL-10, IL-12, INF-γ and TNF-α were quantified by ELISA. Cytotoxicities on lung cancer cell line A549, breast adenocarcinoma cell line MFC-7 and human melanoma cell line HME1 were examined by lactate dehydrogenase release method. The results showed that there were significant differences among different groups. The highest proliferation index on days 10 was observed in group CD28mAb, IL-15 and IL-21(255.3 ± 6.3), which was higher than control group, IL-21+IL-15 group and CD28 mAb+IL-21 group (166.6 ± 13.5, 199.4 ± 15.0 and 228.8 ± 16.6) (P < 0.05). The expression of perforin in CD28 mAb+IL-15 group was higher than the other groups. The expression of perforin,GranB and CD107a of costimulatory groups was higher than control group. The cytotoxicities of CD28 mAb+IL-15 group on A549, MFC-7 and HME1 cells (82.2%, 59.3% and 70.6%) were much higher than that of control group (60.9%, 49.6% and 48.4%) (P < 0.05). The highest IFN-γsecretion was found in CD28 mAb, IL-15 and IL-21 groups. It is concluded that there are significant difference of proliferative capacity, cytokine secretion and cytotoxicity after being activated by different stimulatory factors. Adding corresponding stimulatory factors into the culture system displays a great value for target cells culture.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytokine-Induced Killer Cells ; cytology ; drug effects ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-15 ; pharmacology ; Interleukins ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore the function of a novel nonsense mutation c.1476G>A of ITGB3 gene using an in vitro expression system.

METHODSAn eukaryotic expression vector containing ITGB3 c.1476G>A cDNA was generated by site-directed mutagenesis and transformed into E.coli. Plasmid DNA was extracted and sequenced to confirm the target mutations. Wild-type and mutant recombination plasmids were transfected into Chinese hamster ovarian cancer (CHO) cells by nonliposome method, and the stable expression cells were harvested by G418 screening. The ITGB3 gene mRNA transcription and GPIIIa expression level in CHO cells were detected with real-time quantitative PCR, Western blotting and flow cytometry, respectively.

RESULTSThe eukaryotic expression vectors of wild ITGB3 cDNA and c.1476G>A mutant were successfully constructed. CHO cells with stable expression were obtained after transfection and screening. Compared with the wild-type transfected cells, the amount of CD61 antigen expression was 37% and mRNA transcription level was only 6% in the mutant-transfected cells. Full length GPIIIa protein was found only in the stably wild-type-transfected cells, but not in mutant-transfected cells by Western blotting analysis.

CONCLUSIONThe ITGB3 c.1476G>A mutation can decrease the transcription level and further affect GPIIIa synthesis and CD61 antigen expression.

Animals ; Base Sequence ; Blood Platelets ; cytology ; metabolism ; CHO Cells ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Cricetinae ; Cricetulus ; Humans ; Integrin beta3 ; genetics ; metabolism ; Molecular Sequence Data ; Plasmids ; genetics ; metabolism ; Point Mutation

DNA Polymerase gamma ; DNA-Directed DNA Polymerase ; genetics ; Diffuse Cerebral Sclerosis of Schilder ; diagnosis ; drug therapy ; etiology ; genetics ; Heterozygote ; Humans ; Infant ; Male ; Mutation

BACKGROUNDUnstable pelvic fractures are complex and serious injuries. Selection of a fixation method for these fractures remains a challenging problem for orthopedic surgeons. This study aimed to compare the stability of Tile C pelvic fractures fixed with two iliosacral (IS) screws and minimally invasive adjustable plate (MIAP) combined with one IS screw.

METHODSThis study was a biomechanical experiment. Six embalmed specimens of the adult pelvis were used. The soft tissue was removed from the specimens, and the spines from the fourth lumbar vertebra to the proximal one-third of both femurs were retained. The pubic symphysis, bilateral sacroiliac joints and ligaments, bilateral hip joints, bilateral sacrotuberous ligaments, and bilateral sacrospinous ligaments were intact. Tile C pelvic fractures were made on the specimens. The symphysis pubis was fixed with a plate, and the fracture on the posterior pelvic ring was fixed with two kinds of internal fixation in turn. The specimens were placed in a biomechanical machine at a standing neutral posture. A cyclic vertical load of up to 500 N was applied, and displacement was recorded. Shifts in the fracture gap were measured by a grating displacement sensor.

STATISTICAL ANALYSIS USEDPaired-samples t-test.

RESULTSUnder the vertical load of 100, 200, 300, 400, and 500 N, the average displacement of the specimens fixed with MIAP combined with one IS screw was 0.46, 0.735, 1.377, 1.823, and 2.215 mm, respectively, which was significantly lower than that of specimens fixed with two IS screws under corresponding load (P < 0.05). Under the vertical load of 500 N, the shift in the fracture gap of specimens fixed with MIAP combined with one IS screw was 0.261 ± 0.095 mm, and that of specimens fixed with two IS screws was 0.809 ± 0.170 mm. The difference was significant (P < 0.05).

CONCLUSIONThe stability of Tile C pelvic fractures fixed with MIAP combined with one IS screw was better than that fixed with two IS screws.

Adult ; Bone Screws ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; surgery