ABSTRACT
Background N-nitrosamines, a group of by-products of drinking water disinfection, have strong cytotoxicity to mammals. N-nitrosamines in drinking water are at the ng·L−1 level, and its accurate qualitative and quantitative analysis is difficult, so it is necessary to develop a sensitive and accurate method to determine N-nitrosamines in drinking water. Objective To establish a solid phase extraction-gas chromatography tandem mass spectrometry (GC-MS/MS) method for simultaneous determination of 10 kinds of N-nitrosamines in drinking water. To apply the established method to determine the levels of 10 kinds of N-nitrosamines in drinking water in Nanjing, and to understand the pollution status. Methods Coconut charcoal solid phase extraction (SPE) cartridge and HLB Pro SPE cartridge were compared for the extraction efficiency of 10 N-nitrosamines in drinking water. A coconut charcoal SPE cartridge and a HLB Pro SPE cartridge were concatenated using a SPE connector, and then formed two combinations: coconut charcoal (top)-HLB Pro (bottom) and HLB Pro (top)-coconut charcoal (bottom), to extract the spiked samples, and combined with direct and independent elution ways to obtain the best extraction efficiency. From November to December 2021, 9 raw water, 10 finished water, and 7 tap water samples were collected from 9 municipal water supply units in Nanjing with 1 L brown glass sampling bottles. An 1.0 L drinking water sample was added with the isotope internal standard to prepare a test sample containing an isotope internal standard concentration of 25 ng·L−1. The automatic SPE instrument loaded all the 1.0 L drinking water samples to the tandem SPE cartridge of the HLB Pro (top)-coconut charcoal (bottom) at the rate of 15 mL·min−1. After extraction, the HLB Pro SPE cartridge and coconut charcoal SPE cartridge were transferred to the solid phase extraction vacuum device and eluted with 10 mL of dichloromethane respectively, then the dichloromethane eluents were combined, and concentrated to about 1.0 mL by nitrogen blowing after a small amount of the upper aqueous phase was removed. The concentrated solution was detected by GC-MS/MS and quantified by isotope internal standard method. Results The comparison of sample spike recovery experiments showed that coconut charcoal solid phase extraction (SPE) cartridge and HLB Pro SPE cartridge presented highcomplementarity for the extraction efficiency of 10 N-nitrosamines in drinking water. Using HLB Pro (top)-coconut charcoal (bottom), independent elution, and combined with eluents, the optimal extraction efficiency was obtained. Under these conditions, by GC MS/MS, the 10 N-nitrosamines showed a good linear relationship within the range of 2–50 ng·L−1, the correlation coefficients were all greater than 0.9996, the method detection limit was 0.149–0.211 ng·L−1, and the limit of quantification was 0.596–0.844 ng·L−1. At the spiked concentrations of 5.0, 15, and 30 ng·L−1, the average recoveries of the 10 kinds of N-nitrosamines were 88.0%–104.8%, and the relative standard deviations were 1.22%–4.87%. When applying the method to determine the concentrations of the 10 N-nitrosamines in Nanjing drinking water, the results showed that the 10 N-nitrosamines were positive in different degrees in raw water, finish water, and terminal water, the detection rates were 0%–100%, and the concentrations were ND–27.6 ng·L−1. Conclusion This tandem solid phase extraction-gas chromatography tandem mass spectrometry method can achieve simultaneous determination of a variety of N-nitrosamines in drinking water with high sensitivity and high throughput.
ABSTRACT
Background N-nitrosodimethylamine (NDMA), a new disinfection by-product in drinking water, has attracted extensive attention due to its high detection rate and concentration. Objective To investigate the concentration of NDMA in drinking water in Nanjing situated in the lower Yangtze River Basin of China, and to evaluate associated human health risk. Methods In dry period (January–March) and wet period (July–September) of 2021, raw water, finished water, and tap water from 16 centralized water treatment plants in Nanjing were collected to detect the concentration of NDMA in water samples by solid phase extraction and gas chromatography-triple quadrupole mass spectrometry with programmable temperature vaporizer-based large volume injection. The concentrations of NDMA in water samples of different water types, water periods, and disinfection methods were analyzed, and the daily exposure levels and carcinogenic risk values of NDMA in drinking water of different exposure routes and different exposed populations were calculated. Monte Carlo simulation was implemented with Crystal Ball 11 software to establish a health risk assessment model and conduct sensitivity analysis. Results A total of 61 drinking water samples were collected in Nanjing, and NDMA was positive in all the water samples, with concentrations ranging from 1.36 to 25.65 ng·L−1 and an overall average concentration of (8.00±4.06) ng·L−1. There were no statistically significant differences in the average concentrations of NDMA among raw water, finished water, and tap water samples (F=2.875, P=0.064), between wet season and dry season (t=−0.855, P=0.397), or among different disinfection methods by liquid chlorine, sodium hypochlorite, and chlorine dioxide (F=0.977, P=0.385). The results of health risk assessment showed that the average carcinogenic risk of NDMA and its P95 were 5.95×10−6 and 1.12×10−5 respectively for oral intake of drinking water, and the values for dermal contact were both lower than 1.00×10−6. The mean carcinogenic risks of exposure to drinking water NDMA in children, adolescents, and adults were 1.84×10−6, 8.27×10−7, and 3.28×10−6, respectively. The results of sensitivity analysis showed that the contributions of daily drinking water volume and NDMA concentration in drinking water to the calculated health risk were high, and the contribution of body weight was negative. Conclusion There is a potential carcinogenic risk of NDMA in the drinking water of Nanjing section of the lower Yangtze River Basin，but it is within the acceptable range.
ABSTRACT
Objective:To investigate the risk factors for wheezing in the children with HAdV pneumonia.Methods:Ninety-seven cases of children aged 2 to 5 years with HAdV pneumonia were selected from Pediatric Department of Shengjing Hospital of China Medical University from January 2019 to December 2019.Meanwhile, 100 children of the same age without adenovirus pneumonia were recruited as the control group.According to whether wheezing or not, the children with HAdV pneumonia were divided into wheezing group and non-wheezing group.The general characteristics, clinical characteristics and laboratory examination data of HAdV pneumonia group and non-HAdV pneumonia group were statistically analyzed, and the risk factors of wheezing after HAdV pneumonia were analyzed by multivariate logistic regression.Results:Compared with the non-HAdV group, fever duration, serious cases, cases of wheezing in HAdV group were significant different( P<0.05). In laboratory examination, the white blood cell(WBC)count in HAdV group was significantly higher than those in control group( P<0.05). Whereas, hospitalization time, C-reactive protein(CRP), alanine aminotransferase(ALT), creatinine(Cr), creatine kinase(CK), CKMB, and lactic dehydrogenase(LDH)levels in HAdV group were not statistically significant compared with those of the children in non-HAdV pneumonia group( P>0.05). Wheezing occurred in 52.6%(51/97)of children with HAdV pneumonia.Compared with the non-wheezing group, history of wheezing, cases with atopy, serious cases, eosinophils(EO)count, and cases with small airway changes in wheezing group were significantly different( P<0.05). Whereas, atopic family history, hospitalization time, fever duration, lymphocyte(LY)count, WBC count, CRP level, the cases with co-infection and consolidation in lung images in wheezing group were not statistically significant compared with those of the children in non-wheezing group( P>0.05). Multivariate logistic regression analysis showed that the risk of wheezing in children with severe HAdV pneumonia was 2.949 times as much as those without severe HAdV pneumonia.The risk of wheezing in children with HAdV pneumonia with atopic constitution was 3.930 times as much as those without atopiy. Conclusion:The incidence of wheezing in children with HAdV pneumonia is higher than those in children of non-HAdV pneumonia.Severe cases and atopy are the independent risk factors for wheezing in the children with HAdV pneumonia.
ABSTRACT
Human adenovirus(HAdV)is one of the most common pathogens in children with community-acquired pneumonia.Due to the heterogeneity of the lesion site of adenovirus pneumonia and the easy involvement of small airways, it is often associated with a wheezing attack, which can also lead to an acute attack of asthma, or even an independent risk factor for bronchial asthma.Childhood adenovirus infection can often lead to latent infection and long-term carrying, and some children can be left with different degrees of pulmonary sequelae.In recent years, the number of children with wheezing after HAdV infection has increased gradually.As an important pathogen leading to asthmatic diseases in children, HAdV has been paid more and more attention in clinical practice, and it is particularly important to explore its pathogenesis.
ABSTRACT
Objective:Intensive physicians have relatively insufficient knowledge and experience in treating patients with decompensated schistosomiasis cirrhosis (DSC) admitted in intensive care unit (ICU), but are relatively familiar with patients with decompensated alcoholic cirrhosis (DAC). For this purpose, the clinical characteristics and prognosis of these patients were compared and analyzed.Methods:A retrospective analysis was performed from January 2013 to May 2019 in our hospital and Quzhou People’s hospital. The demographic data, laboratory examination, liver function, Child-Pugh classification, complications of cirrhosis, ultrasonic imaging gastroscopy manifestations were recorded and analyzed. In addition, the treatments and prognosis were also compared.Results:A total of 30 patients (12 males and 18 females) with DSC (aged 57-88) and 31 patients with DAC (aged 41-75) were collected. Compared with patients with DAC, DSC patients were more likely to have coronary heart disease, lower proportion of hyponatremia and lower need of ventilator support. Although the incidences of jaundice and hepatic encephalopathy were significantly reduced ( P<0.05), but parameters of liver function and coagulation were no significant differences in both groups ( P>0.05). B-mode ultrasound of liver in patients with DSC displayed more proportion of patchy and diffuse echo changes and liver volume reduction ( P<0.05), whereas the manifestations of gastroscope in both groups were similar. No significant difference in main treatment measures like uses of somatostatin and three-chamber and two-capsule tube was observed. After treatment, the stop time of gastrointestinal bleeding was similar between groups of DAC and DSC [1.25 (0.5-4.125) days vs. 1.75 (1-2.375) days] ( P>0.05). In addition, the length of ICU stay in DAC group was similar to DSC group [(4.96±3.58) days vs. (3.82±1.99) days], so did the 28-day mortality [14.29% (2/14) vs. 18.18% (2/11)] (both P>0.05). Conclusions:In genenal, patients with decompensated schistosomiasis cirrhosis have the similar clinical characteristics, major biochemical indicators and accessory examination results like ultrasound and gastroscopic examinations to patients with decompensated alcoholic cirrhosis. After timely treatments, both of these patients could achieve a good prognosis.
ABSTRACT
Objective: To explore the psychological and behavioral characteristics of problematic social networks usage. Methods: Totally 11 problematic social networks users were selected from 269 Beijing college students who often used social networks, and they were also deeply interviewed. The Interpretative Phenomenological Analysis (IPA) was used to analyze transcriptional texts. Results: Problematic social networks usage was classified into the following 6 aspects, deviant behaviors, poor attention state, impaired social function, bad moods, abstinence reaction (withdrawal) and tolerance. Conclusion: The present study generalizes the psychological and behavioral characteristics of problematic social networks usage and offers the basis of exploring its measurement tools.
ABSTRACT
Objective To evaluate the diagnostic efficiency of Q?SPECT, CTPA, Q?SPECT/CT, and Q?SPECT/CTPA for pulmonary embolism (PE) in rabbit models. Methods (1) The PE models were constructed by injecting Gelfoam into the femoral vein of New Zealand rabbits ( n=30) . Q?SPECT, CTPA, Q?SPECT/CT and Q?SPECT/CTPA fusion images were obtained by integrated SPECT/CT. (2) All images were interpreted by two experienced nuclear radiologists who were blind to pathologic findings. The locations and numbers of lung lobes with PE were recorded respectively. ( 3) Serial sectioning of the lungs was per?formed and pathologically determined. (4) Se, Sp and Ac of different methods were compared using McNemar test;PPV and NPV were compared usingχ2 test. Kappa test was used to analyze the consistency between two nuclear radiologists. Kappa values0.75 as good consistency. Results (1) Histologically confirmed emboli were present in a total of 26 pulmonary lobes and absent in 79 lobes. (2)The Se, Sp, Ac, PPV, and NPV of 4 imaging methods were:53.8%(14/26), 93.7%(74/79), 83.8%(88/105), 14/19, 86.0%(74/86) for Q?SPECT;73.1%(19/26), 96.2%(76/79), 90.5%(95/105), 86.4%(19/22), 91.6%(76/83) for CTPA;76.9%(20/26), 93.7%(74/79), 89.5%(94/105), 80.0%(20/25), 92.5%(74/80)for Q?SPECT/CT;88.5%(23/26), 91.1%(72/79), 90.5%(95/105), 76.7%(23/30), 96.0%(72/75) for Q?SPECT/CTPA. (3) McNemar test showed Q?SPECT/CT and Q?SPECT/CTPA had higher diagnostic Se for the detection of PE than Q?SPECT (χ2=4.167, 7.111, both P0.05) . Q?SPECT/CT had higher diagnostic Ac than Q?SPECT (χ2=4.167, P0.05). (4)Kappa values of 4 imaging methods for radiologist 1 and 2 were 0.902, 0.915, 0.973, and 0.884. Conclusions Q?SPECT/CT imaging provides good Se and Sp. The diag?nostic efficiency of Q?SPECT/CT is better than that of Q?SPECT and is corresponded roughly to the efficien?cy of CTPA, Q?SPECT/CTPA. The diagnosis of two radiologists on Q?SPECT/CT images has the best con?sistency.
ABSTRACT
<p><b>OBJECTIVE</b>To elucidate the role of transforming growth factor-beta1(TGF-β1) in epithelial-mesenchymal transition of mesothelial cells and peritoneal metastasis of gastric cancer.</p><p><b>METHODS</b>HMrSV5 cells, a human peritoneal mesothelial cell line, were incubated with TGF-β1, and their morphological changes were observed by phase contrast microscopy. Expressions of α-smooth muscle actin (α-SMA), vimentin, cytokeratin, E-cadherin, phosphorylated-Smad2 and Smad2 were examined by Western blotting. After fibroblastic-like mesothelial cells were co-incubate with HSC-39 cells(gastric cancer cell line), the adhesion and invasion potential of HSC-39 were evaluated by adhesion and invasion assay in vitro.</p><p><b>RESULTS</b>Few mesothelial cells converted to spindle fibroblast-like morphology for 24 h, and remarkable phenotypic changes were observed at 72 h of TGF-β1 activation. TGF-β1 could induce α-SMA and vimentin expression, and down-regulate cytokeratin and E-cadherin expression in mesothelial cells (P<0.05). TGF-β1 induced phosphorylation of Smad2 within 15 min of stimulation, reached a maximum at 30 min after treatment and remained high level during the experiment without affecting total Smad2 expression(P>0.05). The percentage of HSC-39 gastric cancer cells adhered were significantly increased as compared to the control. When the mesothelial cells were treated by TGF-β1 for 72 h, the increased adhesion percentage was(146±17)%(P<0.05). After fibroblastic-like mesothelial cells co-incubated with HSC-39 cells for 48 h, more cancer cells [(61.1±11.4) cells/view field] invaded the coated membrane as compared to the control group [(31.9±8.1) cells/view field] (P<0.05).</p><p><b>CONCLUSION</b>TGF-β1 can induce the transition of mesothelial cells into myofibroblasts and Smad2 signal pathway may play a role in this transition, which is associated with increased adhesion and invasiveness of gastric cancer cells, and provides favorable environment for the dissemination of gastric cancer.</p>
Subject(s)
Humans , Cadherins , Cell Line, Tumor , Epithelial Cells , Epithelial-Mesenchymal Transition , Epithelium , Fibroblasts , Peritoneal Neoplasms , Signal Transduction , Smad2 Protein , Stomach Neoplasms , Transforming Growth Factor beta1 , VimentinABSTRACT
Objective To investigate the value of heart-type fatty acid-binding protein in the diagnosis of early myocardial infarc-tion.Methods In 186 cases of suspected acute myocardial infarction due to chest pain,chest tightness for 3 h,plasma CK-MB, troponin-I(CTn-I)and heart-type fatty acid-binding protein(H-FABP)were detected.The sensitivity and specificity in diagnosing early myocardial infarction were compared among 3 kinds of indexes.Results Compared with the non-infarction group and the con-trol group,plasma CK-MB,CTn-I and H-FABP in the acute myocardial infarction group were significantly increased (P <0.05 );compared with CK-MB and CTn-I,the sensitivity of H-FABP to the diagnosis of acute myocardial infarction within 3 h was higher, but its specificity was lower than that of CTn-I and higher than that of CK-MB.Conclusion For the patients with acute myocardial infarction within 3 h after onset,detecting H-FABP can increase the diagnostic rate of early myocardial infarction to a certain extent.
ABSTRACT
Objective:To construct recombinant lentiviral vectors harboring interference RNA ( RNAi ) targetting murine TNF-αgene,so as to lay the foundation on the RNAi gene therapy.Methods: Three small interfering RNA ( siRNA) sequences targeting murine TNF-αgene ( siRNA1,siRNA2,siRNA3) and negative-control siRNA were designed and synthesized.The inhibition effects of siRNAs on TNF-α,IL-1βand IL-6 secretion of LPS-stimulated RAW264.7 macrophages were observed using real-time PCR and ELISA methods.DNA oligo was designed and synthesized according to the most effective siRNA 2 sequence.The recombinant lentiviral shuttle plasmid expressing short hairpin RNA ( shRNA) was constructed and sequenced.The lentiviral shuttle plasmids with packaging plasmids were transfected into 293T cells to produce lentiviral particles.Results: ①The TNF-αmRNA relative expression levels of siRNA1, siRNA2 and siRNA3 were 0.24±0.01,0.16±0.02,0.19±0.01 respectively,significantly lower than that of negative control (0.95± 0.02) (F=531.3,P0.05).②The TNF-αprotein expression levels of siRNA1,siRNA2 and siRNA3 were (23.95±1.21),(17.27±1.46),(19.07± 1.57)ng/ml respectively,significantly lower than that of negative control (35.37±2.93)ng/ml (F=18.1,P=0.000 6<0.001).The inhibition rates of protein expression were 32.29%, 51.16%, 46.08%, respectively comparing with negative control.③The PCR product electrophoresis showed that recombinant vectors yielded 343 bp fragments,non-constructed vectors yielded 306 bp fragments.DNA sequencing partially showed insertion sequence.④Lentiviral particles were obtained by transfecting 293T cells with recombinant lentiviral shuttle plasmids and lentiviral packaging plasmids.Cells grew well during virus production with strong fluorescence expression.The titer of concentrated virus was 2×106 TU/μl.Conclusion:The lentiviral vector harboring RNAi targeting murine TNF-αgene has been successfully constructed.
ABSTRACT
Objective To evaluate the relationship between the level of AMP-activated protein kinase (AMPK) in hippocampus and cognitive dysfunction after splenectomy in aged rats.Methods Sixty-three male aged Sprague-Dawley rats,weighing 480-550 g,were randomly divided into 3 groups (n =21 each):control group (group C),anesthesia group (group A),and surgery group (group S).Morris water maze (MWM) test was performed to evaluate the spatial learning and memory ability before surgery and at 1,3 and 7 days after surgery.The escape latency and swimming distance were recorded.Seven rats were chosen after MWM test was performed at 1,3 and 7 days after surgery and sacrificed.Their hippocampi were removed for detection of the expression of AMPK and phosphorylated (p-AMPK) (by Western blot).Results Compared with the baseline,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery in group S (P ＜ 0.05).Compared with group C,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery,the expression of AMPK was up-regulated at 1,3 and 7 days after surgery,and the expression of p-AMPK was up-regulated at 1 and 3 days after surgery in group S (P ＜ 0.05),and no significant changes were found in the indices mentioned above in group A (P ＞ 0.05).Conclusion Increased AMPK level in hippocampus is the regulatory mechanism of the body adapting to the development of cognitive dysfunction after splenectomy in aged rats.
ABSTRACT
Objective To explore clinical application of the combined therapy of manual small incision cataract surgery (MSICS) with intraocular lens (IOL) implantation and trabeculectomy for patients with refractory glaucoma and coexisting cataract.Methods In our hospital,68 patients (76 eyes) with angle-closure glaucoma of continuously high intraocular pressure (lOP) and coexisting cataract were included in this study from April 2008 to October 2011.All of them were treated with the combination of MSICS with IOL implantation and trabeculectomy.Postoperative visual acuity,IOP,anterior chamber depth (ACD),filtering bleb,ocular fundus and complications were observed.Results Each operation was successfully completed.Postoperative visual acuity and IOP were significantly improved and decreased,respectively,compared to the preoperative.ACD was deepen and filtering bleb was effectively established in the postoperative eyes without significant complications.Conclusion The combined surgery of MSICS with IOL implantation and trabeculectomy might effectively increase visual acuity and decrease IOP,which was a safe and effective approach for patients with refractory glaucoma and coexisting cataract.
ABSTRACT
Objective To explore the deficit features of verbal working memory of dyslexia children by comparing with normal children. Methods Seventeen normal children and fifteen dyslexia children finished the classic verbal n-back task which involved three memory updating difficulty levels. Results Dyslexia children ((81.7±7.0)%,(70.8±12.0)%,(60.0±7.0)%)performed worse than NC ((88. 6 ±6. 0)% , (83. 5 ± 7.0)% ,(75.6 ±9.0)%)in all memory updating levels(n = 0,1 ,2) (P<0.01). In the performances related to memory updating operating,when the difficulty of the updating rised dyslexia children(25. 9 ± 12. 0)% revealed greater decline than normal children(14.2 ± 12.0)% (P<0.01). Conclusion Dyslexia children demonstrate the deficit in the basic level of working memory, which was showed as the bad performance of n-back task in updating level zero, and also they had problems in memory updating,which was showed as the greater updating effect. Memory updating was regarded as the advanced procession of verbal working memory.
ABSTRACT
Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.
ABSTRACT
Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.
ABSTRACT
Objective To evaluate the status of lymph node micrometastases in "non-metastatic" No11P lymph nodes as judged by conventional pathology in the lower third of gastric cancer. Methods In this study 43 No11P lymph nodes harvested from 43 patients which was histologically free of metastasis were examined by consecutive sections and TRAP( telomeric repeat amplification protocol)-ELISA (enzyme linked immunosorbent assay). The data were statistically analyzed according to the clinicopathological features of the patients. Results Micrometastasis was discovered in 4 lymph nodes from 4 patients by consecutive sections. The micrometastatic rate of the conventional pathologic non-metastatic No11P lymph nodes was 9%. The micrometastatic rate of the conventional pathologic non-metastasis No11P lymph nodes detected by TRAP-ELISA was 44%, including 4 lymph nodes observed by consecutive sections It revealed that lymph nodes micrometastases were correlated with the size of the tumor( x2 = 8. 488, P < 0. 05 )、and tumor stage (x2 = 12. 022,P < 0. 05 ). It also showed that the micrometastatic rate increased proportionally to tumor infiltration depth(x2 =6. 473, P <0. 05), not correlated with patients' demographic features, general type and histological differentiation of the tumor. Conclusions There was a high rate of micrometastasis in No11P lymph nodes. This lymph nodes micrometastasis was correlated with the size of the tumor, invasion depth of primary tumor and patients' clinical stage.
ABSTRACT
Objective To establish a UV spectrophotometry method for determination of diphenylamine in air of workplaces.Methods The air samples were collected by glass fiber filter treated with sulfuric acid,desorbed with alcohol by ultrasonic for 10 min,determined by UV spectrophotometry.Results The detection range of the method was 0.19-60 mg/m3(the air volume was15 L).The regression equation:y=0.13x-0.016.The correlation coefficients was 0.999 9.The detection limit was 0.28 ?g/ml.The relative standard deviations was 1.9% -8.4%.Sample-collected efficiency was 99%,desorption efficiency was 98% and the penetration capacity was more than 2.2 mg.Sample stability was 10 days at least by being stored in a refrigerator at 4℃ or by ambient storage.Conclusion This method is applicable to the determination of diphenylamine in air of workplaces.
ABSTRACT
Objective To develop a method for the determination of DBP and DEHP in cosmetics by GC-MS. Methods The samples was extracted by methanol or acetic ether, DBP and DEHP were separated by GC and determined by MS. Results The linear range, regression equation of calibration curve and correlation coefficient were 0.5-100.0 ?g/ml, y=2?106 x-2?106, 0.999 6 for DBP respectively and 5.5-110.0 ?g/ml, y=988 223 x-7?106, 0.995 9 for DEHP respectively. Based on threefold ratio of signal and noise, as the sample was 1.0 and 0.2 g respectively, the volume was 10.0 ml, the detection limits were 1.0 and 5.0 mg/kg respectively for both DBP and DEHP. The recovery rates were 90.8%-119.0% for DBP and 90.4%-115.3% for DEHP. Relative standard deviations were 4.8%-9.8% for DBP and 6.0%-8.6% for DEHP. Conclusion This method is sensitive, accurate and high reproducible, and was applicable to the determination of DBP and DEHP in the cosmetics.
ABSTRACT
Objective To evaluate the clinical diagnostic criteria of latent autoimmune diabetes of adults(LADA).Methods A total of 268 subjects consisted of 168 patients with type 2 diabetes(T2DM) and of 100 normal controls were involved in the study.The body mass index(BMI) was calculated by formula(kg/m~2) and the levels of fasting blood glucose(FBG),postprandial blood glucose(PBG),fasting C-peptide(FC-P),postprandial Cpeptide(PC-P),GAD antibody(GADA),ICA,IAA and IA-2 antibody were detected. The patients with positive antibody in T2DM group were considered as LADA and the others in T2DM group still as T2DM.Results The GADA,ICA,IA-2 antibody and IAA antibody were detected in 21.18%,14.29%,1.19% and 0.60% of patients with T2DM,respectively.The indexes such as the age of onset ≤ 40 years old,overt symptom,ketosis,without chronic complication and hypertension,BMI90.00%).Conclusion The GADA is the most specific antibody in the diagnosis of patients with autoimmune diabetes.The indexes such as the age of onset≤40 years,overt symptom,ketosis,without chronic complication and hypertension,BMI
ABSTRACT
Objective To clone and express Bacillus thuringiensis subsp. isrealensis(B.t.i.) crystal protein CryIVD gene and determine its mosquito larvicidal activity. Methods The gene encoding CryIVD (2.0 kb or so) was amplified by PCR, the amplified fragment was inserted into E.coli plasmid pUC18 to construct the recombinant cloning and expression vector pUC18 CryIVD, which was named pUC18 1. The ligation was transformed into competent E.coli DH 5? and the recombinant vector pUC18 1 was confirmed by restriction enzyme digestion and DNA sequencing. After being inducted by IPTG, the expression of CryIVD gene in positive clone was detected by SDS PAGE and the mosquito larvicidal activity of CryIVD was also determined by standard bioassay. Results The results showed that the CryIVD gene was successfully cloned and expressed in E.coli DH 5? . Mosquito larvicidal activity of engineered E.coli (LC 50 ) to Cx.pipiens pallens and Ae.albopictus Ⅱ-Ⅲ instar larvae was 2.38?10 6 cells/ml and 1.6?10 7cells/ml respectively. Conclusion The CryIVD gene was successfully cloned and expressed, and a high mosquito larvicidal activity was observed.