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ObjectiveTo resolve the problems related to the abnormal interpretations of real-time fluorescence polymerase chain reaction （PCR） results for tri-allele， to formulate the interpretation methods of real-time fluorescence PCR by referring to multiplex PCR fragment analysis， so as to obtain an accurate， simple and cheap detection method for ABCB1 tri-allele. MethodsA total of 2 794 DNA samples were collected from Xi'an Mental Health Center from March 2020 to March 2021， and 5% of which were selected as experiments. Real-time fluorescence PCR method and multiplex PCR fragment analysis method were performed respectively. According to the comparison of Ct values of PCR curves and the comparison of base peak intensity in multiplex PCR fragment analysis， comparison and analysis were conducted on the interpretation results of the two methods， and samples with different interpretation results were verified， thereafter， PCR interpretation method was formulated. ResultsA total of 139 samples were collected， of which 120 alleles and 19 tri-allele were detected. The results of allele detection by two methods were absolutely consistent. In combination with the results of multiplex PCR fragment analysis， a method for the interpretation of real-time fluorescence PCR for 19 cases of tri-allele was developed as follows： when ∣∣Ct.G－Ct.T∣－∣Ct.G－Ct.A∣∣ in amplification curve was less than 3， the interpretation results were the combination of the base pairs with small Ct values； when ∣∣Ct.G－Ct.T∣－∣Ct.G－Ct.A∣∣ was greater than or equal to 3， the interpretation results were homozygotes from the base pairs with minimum Ct values. According to the interpretation method， the results of real-time fluorescence PCR were revised， and it showed 1 case of G/G， 1 case of A/A， 4 cases of T/G， 5 cases of T/A and 8 cases of T/T， which were consistent with the results of multiplex PCR fragment analysis. ConclusionReferring to the multiplex PCR fragment analysis method， the interpretation of ABCB1 tri-allele by real-time fluorescence PCR is developed， and the two interpretation methods are in good agreement.
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The diagnosis of food allergy in children is one hotspot attracting people′s attention in recent years.The incidence of it shows an increasing trend which exposes problems in the understanding of children′s food allergy in China, especially in the misdiagnosis and missed diagnosis.To further standardize the diagnosis and treatment of food allergy in children, based on the current domestic, foreign guidelines and relevant research evidence, the guideline recommends 16 clinical hot-button issues in the 4 aspects of diagnosis, treatment, prognosis, and prevention.Finally, a diagnosis flowchart has been formulated.The guideline aims to improve the standard diagnosis and treatment of food allergies in children in China.
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Objective To describe a case of lower limbs infection of Helcococcus Kunzii in North China University of Science and Technology Affiliated Hospital in October 2017 and to analyze the etiology and drug susceptibility??Methods The bacteria were identified by French Meriere mass spectrometer and BD Phoenix?100 automatic bacterial identification／drug sensitivity system??The drug resistance of the bacteria was detected by disk diffusion method,and collected the clinical information and related literature information of the Helcococcus Kunzi to analyze??Results The Gram?positive cocci was isolated from the foot secretion of a patient with venous thrombosis of lower limbs complicated with infection??The result of smear was Gram?positive cocci in double,agglomerate and varied in size??On the blood plate,there was a small colony of α hemolysis??Thixozyme negative,β galactosidase negative??The results of mass spectrometry identification and two kinds of fully automatic bacterial identification ／ drug sensitivity system were identified as Helcococcus Kunzii,the isolates was resistant to erythromycin and clindamycin among the 10 antibiotics tested??By searching,sorting out and analyzing the literature information, it is revealed that the infection of this bacterium is not related to sex??Middle?aged and elderly people are mostly infected??And the cocci easily infected patients with lower immunity??The infection of lower limbs is 47??37%,the infection of blood flow is 15??79% and the other ways of infection are 42??11%( lung, mammary gland, etc??)??Conclusion Helcococcus Kunzii is a conditional pathogen, and patients with lower immunity are more common??Erythromycin resistance could identify it from the green balloon bacteria??Moreover, the bacteria could cause a wide spectrum of diseases,and easily cause sepsis and lower limbs infections,which reminds us that we should be vigilant in clinical practice??
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Objective To investigate clinical manifestations,pathological features and outcomes of primary osteoma cutis in children.Methods Eleven children with confirmed primary osteoma cutis diagnosed in Department of Dermatology,Beijing Children's Hospital between 2011 and 2018 were included into this study.The clinical manifestations,histopathological features,and outcomes were analyzed retrospectively.Results Among the 11 patients,7 were males and 4 were females.Primary osteoma cutis occurred within 22 months after birth in all the children,the median age of onset was 1 month,and the disease occurred during the first 6 months of life in 10 children.The skin lesions were characterized by skincolored or reddish indurated papules,plaques or nodules of varying size with slight epidermal atrophy.Three patients had local skin lesions,and 8 had multiple skin lesions.Serum calcium and parathyroid hormone levels were within normal limits in all the children,and no developmental deformity was observed at birth in any of the children.Histopathological examination revealed the formation of mature lamellar bone in the dermis in all the cases,which involved the subcutaneous adipose tissue in 5 cases.The skin lesions became stable 8-18 months after the occurrence in 10 patients,which was consistent with primary plaquelike osteoma cutis.Only 1 patient underwent a slowly progressive course,and the skin lesions involved subcutaneous deep tissues,leading to dyskinesia,which was consistent with progressive osseous heteroplasia.Conclusions Primary osteoma cutis in children mostly occurs in infancy,whose clinical manifestations are atrophic,indurated plaques or nodules,and its main pathological feature is the formation of mature lamellar bone.Long-term follow-up is needed,and attention should be paid to the occurrence of progressive osseous heteroplasia.
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Objective Try to find the new biological compounds, the research on the chemical constituents in the fermentation products of an endophytic fungus Phomopsis fukushii had been carried out. Methods The chemical constituents in this fermentation products were isolated by silica gel, Sephadex LH-20 column chromatographies and RP-HPLC methods. Their structures were elucidated by using various spectroscopic techniques. Results Four pentylated diphenyl ethers (1-4) were isolated from this fermentation products, and the new compound (1) was evaluated for its anti-methicillin-resistant Staphylococcus aureus (anti-MRSA) activity. Compounds 2-4 were identified as diorcinol C, diorcinol D, and diorcinol E. Conclusion Compounds 2-4 are isolated from the fermentation products of endophytic fungus Phomopsis fukushii for the first time. Compound 1 is a new compound named phomodiphenyl ether A and given the system name of 1-[4-(3-hydroxy-5-methylphenoxy)-2-methoxy-6-methylphenyl]-3-methylbut- 3-en-2-one. Compound 1 also shows strong anti-MRSA activity with MIC90 value of (54 ± 4) μg/mL. This valve is close to that of positive control, levofloxacin with MIC90 value of [≥ (56 ± 6) μg/mL].
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Objective To investigate the antimicrobial resistance of Streptococcus pneumoniae strains isolated from multiple medical institutions across Hebei Province in 2015.Methods The bacterial data were collected from 53 member hospitals of Hebei Antimicrobial Resistance Investigation Net (HEBARIN) according to the unified surveillance program in Hebei province.WHONET 5.6 was used to review,analyze and summarize the surveillance data.The results were interpreted according to CLSI guideline 2014.Results A total of 2 408 strains of S.pneumoniae were included in this analysis.S.pneumoniae was the third most frequently isolated gram positive bacteria.More than 95％ of these S.pneumoniae strains were susceptible to vancomycin and moxifloxacin.However,96.4％,89.3％ and 67.4％ of these strains were resistant to erythromycin,clindamycin,and trimethoprim-sulfamethoxazole,respectively.The antimicrobial susceptibility profile was similar between the strains isolated from adults and those isolates from children.Conclusions The antimicrobial resistance profile ofS.pneumoniae isolates in Hebei Province is generally consistent with the nation-wide data,except higher resistance level to a few antimicrobial agents.We should be alert to and control the emergence of resistant S.pneumoniae.
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We have carried out the chemical investigation on the roots of Alangium chinense. The chemical constituents from the roots of A.chinense were isolated and purified by various chromatographic techniques, such as silica gel, MCI-Gel resin, Sephadex LH-20 and high performance liquid chromatography. As a result, three alkaloids (1-3) were isolated from 90% EtOH extracts of the roots of this plant. Their structures were elucidated by physical-chemical properties and spectral data. Among them, compound 1 is a new compound, determined as 8-hydroxy-3-hydroxymethyl-6,9-dimethyl-7H-benzo[de]isoquinolin-7-one. Cytotoxicity of the compounds was evaluated by the MTT method. Compound 1 displayed cytotoxicity against NB4, A-549, SHSY5Y, PC-3 and MCF-7 cell lines with IC₅₀ values of 4.2, 3.5, 5.7, 2.8 and 3.9 μmol•L⁻¹, respectively.
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For the purpose of finding new bioactive agents from ethnic medicines, the chemical study on Dai Medicine Cassia occidentalis was carried out. The chemical constituents from the seeds of C. occidentalis were isolated by column chromatographic methods on silica gel, MCI-Gel resin, Sephadex LH-20, and high performance liquid chromatography. Their structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. The cytotoxicity of the compound for NB4, A549, SHSY5Y, PC3, and MCF7 cells line was also assayed by using the MTT method. Two sesquiterpenes (1 and 2) were isolated from this plant. Compound 1 is a new compound and named as methyl 6-(hydroxymethyl)-4-isopropyl-7-methoxynaphthalene-1-carboxylate. Compound 1 also displayed high cytotoxicity with the tested cancer cell-lines.
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A new isoindole alkaloid (1), has been isolated from the leaves of Cassia siamea by using various chromatographic techniques. Compound 1 is a new compound, determined as 5-(hydroxymethyl)-2-methyl-6-prenylisoindolin-1-one, and it displayed cytotoxicity against NB4, A549, SHSY5Y, PC3 and MCF7 cell lines with IC₅₀ values of 3.2,4.6,2.8,6.4, 2.5 μmol•L⁻¹, respectively.
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A new furan-2-carboxylic acid, 5-[3-(hydroxymethyl)-4,5-dimethoxyphenyl]-3-methylfuran-2-carboxylic acid(1),has been isolated from the bark of Cassia alata by using various chromatographic techniques. It displayed cytotoxicity against NB4, A549, SHSY5Y, PC3 and MCF7 cell lines with IC₅₀ values of 2.5, 1.2, 2.2, 3.6 and 1.9 μmol•L⁻¹, respectively.
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The phytochemistry investigation on the Cassia occidentalis, a Dai Medicine, was carried out. The C. occidentalis was extracted with ethanol and then partitioned with EtOAc. The EtOAc soluble materials were subjected repeatedly to column chromatography on silica gel and preparative RP-HPLC, leading to isolation of a nor-sesquiterpene, 3-isopropyl-1,6-dimethoxy-5-methyl-naphthalen-7-ol (1), and a sesquiterpene, 2,7-dihydroxy-4-isopropyl-6-methyl-naphthalene-1-carbaldehyde (2). Their structures were determined by means of spectroscopic studies. Compound 1 is a new compound. Compound 2 is also isolated from C. occidentalis for the first time. In addition, the cytotoxicity of compound 1 for NB4, A549, SHSY5Y, PC3, and MCF7 cells line was assayed by using the MTT method, and it displayed potential cytotoxicity for the tested cancer cell-line with IC₅₀ valves of (1.8±0.2), (1.2±0.2), (0.9±0.1), (2.2±0.3), (2.6±0.3) μmol•L⁻¹, respectively.
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Objective To investigate clinical characteristics of tuberous sclerosis(TSC)without nervous system abnormalities in infants. Methods Clinical manifestations of 12 infants with TSC were analyzed. Reflectance confocal microscopy (RCM)was performed in 5 of the 12 infants. Two patients underwent histopathological examination and electron microscopic examination. Results The age at first clinic visit for TSC ranged from 2 to 18 months (median, 8.6 months)among these patients. Before occurrence of nervous symptom, TSC mainly manifested as hypomelanotic macules, nodules in the subependymal layer or walls of lateral ventricles of the brain, cardiac rhabdomyoma and multiple renal cysts. RCM revealed intact dermal papillary rings with low reflectivity and obscure boundaries in 5 infants. Histopathological examination showed a local decrease in melanocytes and pigment granules in basal cells. Electron microscopic examination showed a normal number of melanin granules in the cytoplasm of melanocytes in the basal layer. Conclusions Before appearance of nervous system abnormalities, TSC mainly manifests as hypomelanotic macules in infants, and is easily confused with vitiligo in clinic. RCM examination may serve as a useful method to distinguish infant TSC from hypopigmented diseases.
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Cancer, an abnormal cell proliferation resulted from multi-factors,has the highest morbidity and mortality among all the serious diseases. Considerable progress has been made in cancer biology in recent years. Tumor immunology, cancer stem cells (CSCs), autophagy, and epithelial-mesenchymal transition (EMT) have become hot topics of interests in this area. Detailed dissection of these biological processes will provide novel directions, targets, and strategies for the pharmacological evaluation, mechanism elucidation, and new drug development of traditional Chinese medicine.
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Animals , Humans , Antineoplastic Agents, Phytogenic , Drugs, Chinese Herbal , Chemistry , Neoplasms , Drug Therapy , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To explore the characteristics, diagnosis and treatment on traumatic dislocation.</p><p><b>METHODS</b>From April 2000 to August 2010, 12 patients with acute traumatic dislocation of superior tibiofibular joint were treated including 10 males and 2 females with an average age of 30.6 years old ranging from 18 to 60 years. According to Ogden classification, 8 cases were the anterolateral dislocation, 2 were posterior-medial dislocation, and 2 were upward dislocation. All patients had swelling in lateral-inferior of knee, fibular head prominent, fibular head pain, floating feeling in head of fibula. One case was treated by conservative treatment, and the remaining 11 cases by surgical treatment.</p><p><b>RESULTS</b>All patients were followed-up for 10 months to 3 years (means 18 months). Evaluation by Lysholm scoring,the total scores were 95.08 +/- 2.02, involving limbing 4.92 +/- 0.28, support 4.92 +/- 0.28, interlocking 15.00 +/- 0.00, instability 24.58 +/- 0.79, pain 22.50 +/- 1.24, swell 8.50 +/- 0.90, climbing stairs 9.75 +/- 0.62, squatting 4.92 +/- 0.28; 11 cases achieved excellent results and 1 good. Nerve functional recovered. X-ray was no longer dislocation.</p><p><b>CONCLUSION</b>The diagnosis of traumatic dislocation is easy misseddiagnosis, surgery is the main treatment method, the prognosis is good.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Arthrography , Fibula , Joint Dislocations , Diagnostic Imaging , Therapeutics , Joints , Wounds and Injuries , Tibia , Tomography, X-Ray Computed , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To investigate clinical effects of coracohumeral ligament reconstruction with autologous double-strand of long palmaris longus tendon and artificial ligament for the treatment of acromioclavicular joint dislocation.</p><p><b>METHODS</b>From April 2006 to June 2009, 31 patients with acromioclavicular joint dislocation were treated with coracohumeral ligament reconstruction using autologous double-strand palmaris longus tendon and artificial ligament. There were 18 males and 13 females, ranging in age from 18 to 60 years, with an average of 35 years. Twenty-six patients were acute trauma and other 5 patients were chronic trauma. Preoperative symptoms included different degrees of pain, restricted movement, and instability of acromioclaviecular joint. The X-ray showed acromioclavicular joint dislocation.</p><p><b>RESULTS</b>The patients had good incision union without vascular and nerve injuries. All the patients were followed up, and the average duration was 23 months. The JOA scores decreased from preoperative (38.8 +/- 1.5) to (73.2 +/- 1.1) at 1 month after operation,and (93.5 +/- 0.8)at the last follow-up. Twenty-eight patients got an excellent result, 2 good and 1 fair.</p><p><b>CONCLUSION</b>The reconstruction of coracohumeral ligament using autologous double-strand palmaris longus tendon and artificial ligament is an effective method for the treatment of acromioclavicular joint dislocation.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Acromioclavicular Joint , Wounds and Injuries , General Surgery , Artificial Organs , Clavicle , Follow-Up Studies , Joint Dislocations , General Surgery , Ligaments, Articular , General Surgery , Plastic Surgery Procedures , Methods , Scapula , Tendons , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To explore the effect and mechanism of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma SMMC-7721 cells.</p><p><b>METHODS</b>SMMC-7721 cells were treated with different doses of Poly I:C for 24, 48, and 72 h, and the cell growth inhibition rate was analyzed with CCK-8 assay. The cell cycle and the apoptosis were analyzed using flow cytometry with Annexin-V and PI staining, and quantitative RT-PCR analysis were used to detect the expression of TLR3, TRIF, and IFN-beta mRNA in cells.</p><p><b>RESULTS</b>In the cells exposed to Poly I:C at low, moderate, and high doses, the inhibitory rates was the highest in high-dose Poly I:C group, and at a given Poly I:C dose, prolonged exposure resulted in significantly increased cell growth inhibition rate (P<0.05). Flow cytometry showed that Poly I:C induced cell apoptosis in a time- and dose-dependent manner and significantly increased the percentage of G1-phase cells as compared with that in the control group. The mRNA level of TLR3, TRIF, and IFN-beta were also increased following Poly I:C treatment in comparison with the control group.</p><p><b>CONCLUSION</b>Poly I:C can induce significant growth inhibition and apoptosis of SMMC-7721 cells in a dose- and time-dependent manner possibly by causing cell cycle arrest and TLR3 signaling pathway activation that leads to IFN-beta production and cell apoptosis.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Cell Proliferation , Dose-Response Relationship, Drug , Interferon-beta , Genetics , Metabolism , Liver Neoplasms , Pathology , Poly I-C , Pharmacology , RNA, Messenger , Genetics , Metabolism , Receptors, Cholecystokinin , Metabolism , Signal Transduction , Toll-Like Receptor 3 , Genetics , MetabolismABSTRACT
Atopic dermatitis (AD) is a chronic, recurrent, and inflammatory skin disease. It usually occurs in children with a clinical manifestation of severe itching. In recent years the incidence of AD is slowly increasing, which severely impacts the physical and mental health of children. This article summarizes the epidemiology, influencing factors, and management of this diseases.
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Humans , Dermatitis, Atopic , Epidemiology , Therapeutics , PrognosisABSTRACT
To develop a fed-batch fementation process of E. coli TOP10 containing a recombinant plasmid pBAD/HBs Fab. Cells were grown in semi-defined medium at 37 degrees C, and the feed operation using glycerol as carbon source was performed when dissolved oxygen increased. When the target cell concentration reached to 64g/L, arabinose was added to a final concentration of 0.02%. Cells were grown for another 5h with the culture temperature decreased from 37 degrees C to 30 degrees C. In the whole process, cell growth was monitored by measuring OD600 of samples taken at 1/2h intervals and the dissolved oxygen was kept above 30%. After the fementation, E. coli pellets were collected for purification of Fab protein. The specificity of Fab protein was confirmed by Western blot, and binding activity to HBsAg was verified by Dot blot. Cell concentration we got is 96g wet bacteria per liter, the Fab protein is about 6% of total protein of the host, that is 80mg per liter. Stable fermentation parameters were obtained for fermentation to improve productivity of the Fab protein. The Fab protein was produced in the form of soluble biologically active protein, it's better than inclusion bodies from which biologically active protein can only be recovered by complicated and costly denaturation and refolding process.
Subject(s)
Blotting, Western , Escherichia coli , Genetics , Metabolism , Fermentation , Physiology , Hepatitis B Surface Antigens , Allergy and Immunology , Immunoglobulin Fab Fragments , Genetics , Allergy and Immunology , TemperatureABSTRACT
Objectives To determine the prevalence of Staphylococcus aureus (S. Aureus) colonization and S. aureus-derived exotoxins in lesions of childhood atopic dermatitis (AD) and evaluate the role of S.aureus-derived exotoxins in the pathogenesis of childhood AD. Methods Specimens were taken from the skin lesions of 148 patients, non-lesional skin of 30 patients, and the skin of 250 controls for bacterial cultures. S. aureus-derived exotoxins were detected by reverse passive latex agglutination. Total IgE levels were determined with immunoradiometric assay. Results The prevalence of S. aureus colonization was significantly increased in both the lesional and non-lesional skin of patients with AD in comparison with the controls (P 0.05). However, patients with increased total IgE levels showed significantly high SCORAD indices (P
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Objective To study microbial flora on skin lesions and their antim icrobial susceptibilities and to conduct a topical treatment trial in atopic der matitis (AD). Methods Seventy one patients with AD were diagnosed according to the criteria of Hanifin-Rajka′s in departments of dermatology in China-Japan Friendship Hospital and Beijing Children Hospital during November,2001 to March, 2002. Bacteria and fungi were examined by culture and microscopy. Among 71 case s, 66 were divided into two groups randomly with one group receiving 1%econazol e+0.1%triamcinolone acetonide cream topically twice daily for 4 weeks, and the control group receiving 0.1%hydrocortisone butyrate ointment. Severity of AD w as measured by SCORAD index. Results It was shown that AD mainly affected infant s and children with a prevalence of 73.24%in patients less than 12 years old. T he rates of bacterial and fungal colonization were 53.51%, 1.41%, respectively on skin lesions. S.aureus was detected in 35.21%of patients with AD. Antimicro bial susceptibility test indicated that S.aureus was sensitive to rifampin, vanc omycin, amikacin, ciprofloxacin, cefazolin, cefuroxime, etc. It was found that 1 %econazole+0.1%triamcinolone acetonide cream had significantly more advantage over hydrocortisone butyrate ointment in the therapeutical effects (P