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Objective:To establish the HPLC fingerprint of MaRSDenia tenacissima; To evaluate the different origins of MaRSDenia tenacissima by combining chemometric methods.Methods:High performance liquid chromatography (HPLC) method was adopted, with DiKMA C18 column (250 mm× 4.6 mm, 5 μm), acetonitrile-0.05% phosphoric acid aqueous solution as mobile phase gradient elution, flow rate of 1.0 ml/min. The detection wavelength was set at 230 nm, the column temperature was 30 ℃, and sample size was 10 μl. Chromatographic information was imported into the similarity evaluation system for TCM chromatographic fingerprints (2012 version) for similarity analysis. SPSS Statistics 26 was used for system clustering analysis, and SIMCA 14.1 software was used for principal component analysis and partial least squares discriminant analysis (PLS-DA).Results:Totally 12 common peaks were identified. Two chromatographic peaks were identified as tenacissoside G and tenacissoside I. The relative similarity of fingerprints of 15 batches of samples and references ranged from 0.942 to 0.995. When the square Euclidean distance was 20, the samples could be grouped into two categories: S1-S3, S13-S15 were grouped into one category, and S4-S12 were grouped into another category. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that there were significant differences among 15 batches of MaRSDenia tenacissima, and there was a certain correlation with the origin.Conclusion:The results can provide a reference for analyzing the differences of MaRSDenia tenacissima from different producing areas and the quality standards of related formula granules in the later stage.
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Objective:A nested-PCR assay is developed to detect and identify the genomic DNA of Brucella vaccine A19 strain. Methods:The whole genomic sequences of Brucella vaccine A19 strain and other Brucella spp. strains were compared and analyzed. The primers were designed by nucleotide difference sites. The nested-PCR assay was established to detect and identify Brucella vaccine A19 strain. The genomic DNA of Brucella vaccine A19 strain was extracted and diluted. The diluted template DNA was tested for sensitivity of using nested-PCR assay. And the specificity of nested-PCR assay was tested for the genomic DNA of other Brucella spp. strains and non- Brucella spp. strains. Results:The minimum detection limit of the nested-PCR assay was 3.43 fg. The nested-PCR assay established for amplification of Brucella vaccine A19 strain showed 246 bp electrophoresis bands, while other Brucella spp. strains showed 314 bp electrophoresis bands, and non- Brucella spp. strains did not produce electrophoresis bands. Conclusions:The nested-PCR assay established has the characteristics of high sensitivity and specificity. It can be detected when there is one copy of Brucella vaccine A19 strain genomic DNA in the reaction system. This method is particularly suitable for the detection and identification of trace genomic DNA of Brucella vaccine A19 strain in sample.
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Objective:To establish a quantitative real-time PCR detection system for Brucella S2 vaccine strain. Methods:Based on the differences in the entire genome sequence between Brucella S2 vaccine strain and other reference strains of Brucella, primers and probes were designed to establish a quantitative real-time PCR detection system for Brucella S2 vaccine strain. The DNA of 22 reference strains of Brucella and 8 non- Brucella control strains were obtained from the National Institute for Infectious Disease Control and Prevention of the Chinese Center for Disease Control and Prevention. At the same time, environmental samples were obtained from the brucellosis vaccine manufacturers, and bacterial DNA from environmental samples was extracted using a blood/tissue genomic DNA extraction kit. The obtained DNA was pre-amplified by conventional PCR, and then subjected to quantitative real-time PCR secondary amplification (nested fluorescence quantitative PCR) using the amplified PCR product as a template. The specific fluorescence curve and corresponding number of cycles (Ct value) were observed, and the sensitivity was tested. Results:The quantitative real-time PCR detection system established did not detect specific fluorescence curves (without Ct values) for 21 reference strains of Brucella and 8 non- Brucella control strains, except for S2 vaccine strains. The established detection system had a minimum detection limit of 4.34 fg (genomic DNA) for detecting the DNA of Brucella S2 vaccine strain; DNA of Brucella S2 vaccine strain was detected in 3 of the 14 environmental samples collected. Conclusion:The quantitative real-time PCR detection system established can detect Brucella S2 vaccine strain in samples, with good sensitivity and specificity.
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ADP-ribosylation factor 6 (Arf6), a small G-protein of the Ras superfamily, plays pivotal roles in multiple cellular events, including exocytosis, endocytosis, actin remodeling, plasma membrane reorganization and vesicular transport. Arf6 regulates the progression of cancer through the activation of cell motility and invasion. Aberrant Arf6 activation is a potential therapeutic target. This review aims to understand the comprehensive function of Arf6 for future cancer therapy. The Arf6 GEFs, protein structure, and roles in cancer have been summarized. Comprehending the mechanism underlying Arf6-mediated cancer cell growth and survival is essential. The structural features of Arf6 and its efforts are discussed and may be contributed to the discovery of future novel protein-protein interaction inhibitors. In addition, Arf6 inhibitors and mechanism of action are listed in the table. This review further emphasizes the crucial roles in drug resistance and attempts to offer an outlook of Arf6 in cancer therapy.
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OBJECTIVE@#Thinking on the construction of the medical device type archives information system.@*METHODS@#This paper introduces the concept and significance of medical device variety archives, and puts forward the overall construction idea and system framework of medical device variety archives by analyzing its construction difficulties.@*RESULTS@#Considering the long-term nature and complexity of the construction of medical device variety archives, the system can be constructed in accordance with the three steps of system building, platform building and data management, and the overall technical architecture can be designed from the eight aspects of user layer, business application layer, application support layer, data resource layer, infrastructure layer, security, standards and operation and maintenance management.@*CONCLUSIONS@#Architecture design is the foundation of system construction, and its design rationality is very important for the success of system construction. The architecture design proposed in this study has a certain reference role for promoting the construction of medical device variety archives management system.
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Information Systems , Reference StandardsABSTRACT
Objective:To investigate the feasibility and safety of endovascular repair for traumatic thoracic aortic pseudoaneurysm.Methods:From Oct 2015 to Oct 2018, the clinical and followup data of 7 patients diagnosed as traumatic thoracic aortic pseudoaneurysm in Fuwai Hospital of Chinese Academy of Medical Sciences were analyzed retrospectively.Results:The patients average age was (51.2±11.0) years old. All patients underwent surgery in the hybrid operating room under general anesthesia. Two did thoracic endovascular aortic repair (TEVAR), three did TEVAR combined with chimney technique to reconstruct the left subclavian artery, and 1 had TEVAR combined with fenestration to reconstruct the left subclavian artery. One did TEVAR with left common carotid artery and left subclavian artery bypass. The mean operative time was (90.1±27.4) min, the mean postoperative hospital stay was (8.9±3.7) d, and the mean postoperative follow-up time was 42.4 months. All the patients received CTA reexamination of the aorta after 1, 6, 12 months and yearly thereafter. TypeⅠendoleak was found in one patient with chimney technique to reconstruct of left subclavian artery after operation. CT showed that the type Ⅰ endoleak disappeared 6 months after operation. There was no death, paraplegia or stroke during the perioperative period and follow-up period, and there was no aortic related reintervention.Conclusion:TEVAR is a safe and effective method for the treatment of traumatic pseudoaneurysm of thoracic aorta, and the early and mid-term results were satisfactory.
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Objective To compare and analyse the detection performance of different 2019-new coronavirus (2019-nCoV) nucleic acid detection kits, in order to provide references for laboratory. Methods Six kinds of domestic reagents (A—F reagent) were selected for parallel detection of a series of samples from one patient in this hospital whose 2019-nCoV nucleic acid result was confirmed weakly positive. The samples were taken at three different times, the RNAs were extracted and amplified, and two parallel tests were performed each time by use of these six kits. The detection performance was compared according to the results of each kit. Results The three parallel test results (ORF1ab and N gene) of C and F reagents were positive, the results of D reagent showed the N gene was not detected, and the results of A, B, E reagents showed the ORF1ab gene was not detected sometimes. The reproducibility of in-batch detections by C reagent was the best, and the CT values of F reagents (N and ORF1ab), E reagents (ORF1ab) and A reagents (ORF1ab) showed changes in trend. Conclusion There are differences in the detection ability of six 2019-nCoV nucleic acid detection reagents for weakly positive samples, and the accuracy, sensitivity and reproducibility of some reagents are not good. There is an urgent need to further optimize and improve their performance in order to better meet the needs of large-scale screening.
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Objective@#To explore the association between the content and distribution of body fat and early puberty among children and adolescents, and to provide a basis for the study of the mechanism of early puberty.@*Methods@#The questionnaire survey and physical examination were conducted among the students from 2 primary schools with girlsof 3 rd-4 th grade and boys of 4 th-5 th grade and boys and girls in 7 th-8 th grade from 2 middle schools with by purposive sampling in Beijing in early January 2016. Descriptive statistics were used to describe the general information of samples. Logistic regression analyses were used to test the effects of body composition on the early puberty.@*Results@#A total of 1 527 students were included, of which 177 were early puberty and the prevalence of early puberty was 11.6%. The prevalence was 12.2% for girls and 11.0% for boys. The average value of the three skinfold thicknesses of the participants was 15.2±4.8 mm (triceps skinfold), 13.4±6.3 mm (subscapular skinfold), 14.6±6.6 mm (suprailiac skinfold), the average value of the body fat was 22.2 ± 6.2 kg, and the average value of the total fat weight was 11.2 ± 6.2 kg. After adjusting for age, single-child, family economic level and parental education level, multivariate logistic regression showed that girls with high triceps skinfold, subscapular skinfold and suprailiac skinfold were more likely to be early puberty (for triceps skinfold: OR=2.03, 95%CI=1.26-3.27; for subscapular skinfold: OR=2.14, 95%CI=1.32-3.46; for suprailiac skinfold: OR=2.05, 95%CI=1.26-3.31). Body fat content and total fat weight were also the risk factor of early puberty in girls (for body fat content: OR=1.88, 95%CI=1.17-3.02; for total fat weight: OR=2.08, 95%CI=1.31-3.32). For boys, only high subscapular skinfold increased the risk of early puberty(OR=1.90, 95%CI=1.16-3.10).@*Conclusion@#Body fat content and body fat distribution were positive associated with early puberty in children and adolescents, and there are significant gender differences.
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Objective:To investigate the effects of an atypical antipsychotic drug olanzapine on the body mass index(BMI), leptin and hypothalamic neurohormone related to appetite regulation in the first-attack psychotic patients after 4-week treatment. Methods:Totally 38 first-attack psychotic patients meeting DSM-Ⅳ diagnostic criteria were treated with olanzapine for 4 weeks. The BMI,blood lipids,blood glucose and neurohormone were measured before and after the treatment. The normal control group was also considered the above indices. Results:After the 4-week treatment, the body weight and the body mass index increased significantly when compared with those before the treatment (P < 0.01),and the plasma levels of TC,TG and LDL were higher than those before the treatment(P < 0.05). Both leptin and NPY levels significantly increased in the patients after the treatment (P <0.05),while the NPY levels were significantly lower before the treatment when compared with those in the control group(P< 0.05). The α-MSH levels before and after the treatment were significantly lower than those in the control group(P < 0.01). The CART levels had no significant difference among the pre-treatment group, post-treatment group and control group (P >0.05). Conclusion:The levels of leptin and neurohormone may be changed in the early treatment of olanzapine in the patients with schizophrenia,which may cause weight gain.
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Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.
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Objective@#To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression.@*Methods@#UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by 32P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager.@*Results@#UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA.@*Conclusions@#UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.
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Objective To design a scientific, standardized, fully functional, easy-to-use Guillain-Barré syndrome (GBS) clinical informatization management system, with the purpose to meet the needs of diagnosis and clinical research in patients with GBS. This system can manage and excavate clinical data and provide a platform for clinical research of GBS. Methods The clinical data items of GBS were identified. The Visual Basic 6.0 and Vista DB software development environment was used to develop the system and the related database based on embedded database. Results A GBS database including 560 clinical indicators were set up. Through the built-in data mining and analysis functions, the system can realize the functions of visualized data input, combination search, statistical analysis, data exchange and literature update. This system has been used in Xijing Hospital, and a total 274 clinical and prognostic records were recorded from the patients with GBS. The practical results proved that the system can achieve the design goals. Conclusions Based on the information technology and GBS-related clinical epidemiology and neurology expertise, a standard GBS clinical data management and analysis system and related database were established, which could provide a basis for GBS information storage, update, epidemiological investigation and prognosis.
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Macrobrachium nipponensis is delicious and has high economic value, but its susceptibility to white-spot syndrome virus (WSSV) is unknown. Susceptibility, morbidity, and multiplication of WSSV in M. nipponense were studied by epidemiological survey, infection experiment and qPCR. M. nipponense was the natural host of WSSV, and the natural carrying rate was about 8.33%. M. nipponense could be infected with WSSV via oral administration, muscle injection and immersion, and the cumulative infection rate of 10 d exposure was 100%, and the cumulative mortality rates were 100%, 75% and 0%, respectively. The infection of WSSV is fast by muscle injection. The virus content after 5 day's injection is 1 000 times higher than that of the first day of infection, and the mortality rate reached 100% after 8 days. The median lethal dose (LD₅₀) measured as the mortality of infected M. nipponense via injection indicated the LD₅₀ in the concentration of WSSV of 2.71×10⁵ virions/μL. In shrimp farming, M. nipponense can be infected by ingesting WSSV infected shrimp or dead shrimp, and also by soaking in WSSV-containing water and thus become a vector, consequently affecting the spread and pathogenicity of WSSV.
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Objective To determine the association between weight gain induced by atypical antipsychotic and the polymorphisms of MC4R gene rs12970134.Methods 62 patients who had weight gain more than 7% of their pre -drug body weight were selected as study group,and 62 patients who had weight gain less than 7% of their pre -drug body weight were selected as control group.The polymorphism of MC4R gene rs12970134 was analyzed by using polymerase chain reaction and directly sequencing technology.Results There were no significant differences in the frequency of MC4R gene rs12970134 genotypes and alleles between the two groups(χ2 =0.648,P =0.723;χ2 =0.679,P =0.410).While after the treatment with atypical antipsychotic,the weight gain degree in patients with GG genotypes was less than patients with GA /AA genotypes[(22.18 ±0.33)kg/m2 vs.(23.53 ±0.58)kg/m2 ](t =-2.167,P =0.032).Conclusion The polymorphisms of MC4R gene rs12970134 maybe affect the weight gain degree in patients after treatment with antipsychotic.
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Objective To investigate the influence of different intervention models on autism spectrum disorder (ASD) children. Methods Eighty-eight children aged from 12-46 months and newly diagnosed ASD were randomly assigned to different intervention models, including standard intervention group (T1, n=55), non-standard intervention group (T2, n=11), and family intervention group (T3, n=22). The intervention data was recorded including time and methods. Chinese revised version of Psycho-Educational Profile (C-PEP) and social adaptive behavior scale were used to test the development quota?tion (DQ) before and after intervention. Results There were significant statistical differences in C-PEP scale and pathologi?cal score before and after intervention in T1 group (P<0.01). There were significant differences in the pathological score, in?terpersonal and cooperative behaviors, sensory patterns and language barriers after intervention in T2 group (P<0.05). And there were no significant changes in the developmental quotient. The perception, gross motor, cognitive performance and the developmental quotient of oral cognition were significantly reduced after the intervention in T3 group (P<0.05). There was no significant change in pathology score. Results showed that there were significant differences in the imitation, perception, cognitive performance, oral cognition and general development before and after the intervention between three groups ( P<0.05). Conclusion A significant effect is found in children with autism spectrum disorder after standard intervention.
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Aim To explore the inhibitory effects of ophiopogonin-B (OP-B ) on cell adhesion,invasion and migration in non-small cell lung cancer (NSCLC) A549 cells in vitro and its possible mechanism.Meth-ods Cell adhesion assay and transwell chamber assay were used to detect the ability of cell adhesion,migra-tion and invasion.qRT-PCR was used to detect the mRNA levels of MMP-2 and 9 .Meanwhile ,Western blot assay was performed to determine the protein levels of MMP-2/9 and p-Akt.Results OP-B significantly inhibited the ability of cell adhesion,invasion and mi-gration in A549 cells at the concentration of 10 μmol· L-1 (P<0.01 ).Meanwhile,it inhibited the mRNA and protein levels of MMP-2 and MMP-9 and down-regulated the phosphorylation of Akt (P <0.0 1 ). Conclusion OP-B inhibits cell adhesion,invasion and migration in A549 cells through down-regulation of the mRNA and protein expression of MMP-2/9 ,and the inhibitory effect on the expression of p-Akt.
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Aim To study the mechanism of DXM and NAC inhibiting the expression of IL-8 and ICAM-1 in A549 cells. Methods The expression of IL-8 and ICAM-1 was detected by ELISA and flow cytometry re-spectively; the expression of GR,HDAC,AP-1,NF-κB was detected by Western blot, while the activity of HDAC was detected by spectrophotometry. ResultsThe increasing expression of IL-8 and ICAM-1 induced by TNF-α could be inhibited by DXM and NAC in A549 cells. DXM could inhibit the transcribed activa-tion of AP-1,NF-κB, and the expression of HDAC and its activity induced by TNF-α and LPS; NAC only in-hibited the transcribed activation of NF-κB, while it had no affection on the transcribed activation of AP-1 and the expression of HDAC and its activity. Conclu-sions DXM and NAC both have the anti-inflammatory effect. DXM plays the role of anti-inflammation through increasing the expression and activation of HDAC, in-hibiting the transcribed activation of AP-1 and NF-κB, while NAC has no effect on the expression and activa-tion of HDAC, which shows that NAC does not exert anti-inflammatory effect through acetylation signal.
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Objective To investigate the value of Cytochrome P450 (CYP3A5) * 3 gene polymorphism in providing individualized administration for the use of tacrolimus (Tac) in renal transplantation recipients.Method Pyrophosphate sequencing method was used to determine the CYP3A5 * 3 genotype of renal transplant patients in the first day after surgery.Sixty recipients were divided into experiment group and control group.Both groups of patients were routinely given the initial dose of Tac-4.0 mg/day in the first day after surgery.The experiment group of patients were given different doses of Tac based on the different CYP3A5 * 3 genotypes at the third day after surgery [for AA:0.12 mg/(kg· day),and for GG:0.06 mg/(kg· day)],and the control group of patients were given different dosages of Tac according to drug concentration.Different parameters were compared between two groups of patients:percentage of patients reaching the target concentration (3-8 μg/L) at the fifth day after surgery,days required to reach the target concentration level,times needed to adjust the dosage of Tac within two weeks.Result The percentage of patients reaching the target concentration in experiment group and control group was 90% and 46.67%,respectively (P< 0.05).Days required to reach the target concentration were (3.67 ± 1.32) and (7.57 ± 3.42) on average,respectively (P < 0.05).Times of adjusting the Tac dose in experiment group was significantly less than those in the control group (P<0.05).In the experiment group,the target concentration was obtained even without dosage adjustment (70%).Conclusion Individualized adjustment of Tac doses for patients according to recipients' different CYP3A5 * 3 genotypes is beneficial for reaching target concentration as soon as possible,which is superior to traditional dosage regimen.
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Objective To investigate the influence of sevoflurane and total intravenous anesthesia with propofol on hepatic and renal function in elder patients under gastrectomy .Methods 55 patients between the ages of 60-75 ,ASA physical status class Ⅰor Ⅱ ,scheduled for an elective gastrectomy were randomly divided into two groups .Anesthesia was maintained with sevoflurane 1% -1 .2% and remifentanil (0 .1-0 .2)μg · kg -1 · min-1 in the group S and propofol (1 -2)mg · kg -1 · h-1 and remifentanil (0 .1-0 .2)μg · kg -1 · min-1 in the group T .The hepatic and renal function ,aspartate aminotransferase (AST ) ,alanine amin-otransferase(ALT),bloodureanitrogen(BUN)andcreatinineweretestedatpreoperation(baseline),postoperative1dayand3 days .Results AST was increased at postoperative 1 day and 3 day ,compared with that of the preoperation in the group S and group T .Serum BUN at 3 day and creatinine at 1 day and 3 day were significantly higher from the preoperative values in group S (P0 .05) .There were no significant difference in the AST ,ALT , BUN and creatinine between the groups(P>0 .05) .Conclusion The changes of hepatic and renal effect after inhalation anesthesia with sevoflurane and remifentanil and TIVA with propofol and remifentanil for gastrectomy are clinically insignficant ,and there is no difference between the two methods .
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Objective To evaluate the effect of transforming growth factor-β (TGF-β) on the expressions of epithelial-to-mesenchymal transition (EMT)-related molecules in human A431 epidermal squamous cell carcinoma cells.Methods Cultured A431 cells were classified into two groups:TGF-β group treated with TGF-β of 7.5 μg/L for 72 hours,and control group remaining untreated.Subsequently,cell morphology was observed under an inverted microscope,and real time-PCR and Western blot were performed to quantify the mRNA and protein expressions of EMT-related molecules including E-cadherin,N-cadherin,vimentin and β-catenin respectively.Results After TGF-β treatment,the cells became dispersed and spindle in shape.The mRNA expression levels of E-cadherin,Ncadherin,vimentin,and β-catenin in TGF-β group were 0.317,2.475,11.340 and 2.615 folds those in the control group respectively.Western blot showed a marked increase in the expression of N-cadherin,vimentin,and β-catenin proteins but a notable decrease in that of E-cadherin in the TGF-β group compared with the control group.Conclusions TGF-β can induce EMT in A431 cells,and increased expression of TGF-β may contribute to the invasion and metastasis of SCC.