Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P ＜ 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P ＜ 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P ＜ 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P ＜ 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P ＜0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P ＜ 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.

ObjectiveTo explore the normal range of the volume of internal capsules in Chinese adults of the Han nationality and its relationship with age,body habitus,and craniocerebral volume.Methods One thousand healthy volunteers (age range =18 to 80 years) were divided into 5 groups according to their age;Group A ( 18 to 30 years old),group B (31 to 40 years),group C (41 to 50 years),group D (51 to 60 years),and group E (61 to 80 years).Each group consisted of 100 males and 100 females.MR imaging was performed in all of the volunteers using T1 weighted three-dimensional nagnetization prepared rapid acquisition gradient echo sequence. After three dimension data reconstruction,the volumes of bilateral internal capsules were manually measured. The volumes of bilateral internal capsules were compared by paired sample t test.The internal capsule volumes were compared between male and female by independent sample t test,and the differences among 5 age groups were compared by one-way ANOVA.The relationship between the volumes of internal capsule and age,body habitus or cerebral volume were analyzed using bivariate correlation.ResultsThe left and right internal capsule volumes were (2809 ± 393) and (2677 ± 343 ) mm3 respectively.The left internal capsule volumes were significantly larger than that of right (t =12.078,P ＜ 0.05 ).The left and right side of internal capsule volumes in male were (2863 ± 396) and (2744 ±358) mm3 respectively,and (2754 ±385) and (2609 ±314) mm3 in female.The left and right internal capsule volumes were larger in males than in female (t =1.982,2.851 ;P ＜ 0.05 ).The left internal capsule volume of the 5 age groups were ( 3273 ± 361 ),( 2943 ± 299 ),( 2777 ± 255 ),( 2607 ± 199 ),(2444 ±213) mm3,and the right were (2993 ± 361 ),(2814 ± 270),(2682 ± 239),(2543 ± 219),(2351 ±210) mm3.There were significant differences among 5 age groups between left and right internal capsule volume ( F =55.244,34.493 ; P ＜ 0.05 ).There was significant negative correlation between the volume of left and right internal capsule and age ( r =- 0.718,- 0.637 ; P ＜ 0.05 ).Conclusions1.5 T MR scanner can be used to accurately measure the internal capsule volumes.There is a significant negative correlation between age and internal capsule volumes.

Myeloid differentiation factor MyD88 is a critical adaptor molecule that integrates and transduces intracellular signals in inducing the differentiation of dendritic cells (DCs).The domain regions within MyD88 was searched,it could potentially affect the function of dendritic cells and found that MyD88 aa155-171 motif not only regulate the activity of transcription factor NF-?B, but also control the production of cytokines and expression of costimulatory molecules. Indeed, aa155-171 motif deleted type MyD88 (MyD88155-171) transfected RAW264.7 cells exhibited the reduced NF-?B and AP-1 activity and interrupted the expression of CD86 and B7H1. Meanwhile, lower level expression of cytokines such as IL-12,IFN-? were also observed by means of cytokine array in MyD88-/-DC trasfected with MyD88155-171 as compared to the MyD88 transfected cells. Thus, aa155-171 motif inside MyD88 could affect the expression of costimulatory molecules, production of cytokines and transduction of Toll like receptor signal pathway, suggesting that this motif may play an important role in regulating responses of innate immune system.

Destructive injury is defined as a very serious damage both to the bone and the soft tissues. But in clinical practice we found that in some fracture cases, the damage to soft tissues is not as severe as "destructive injury" indicates, whereas comminuted fractures still cannot show the severity of bone damage. Therefore we proposed a new term "destructive fractures" after combining the definition of destructive injury with typical clinic cases. Destructive fractures refer to the fractures whose osseous tissues are damaged too seriously to be repaired, but soft tissues, nerves and veins are less severely injured and can be repaired. From the year 2001 to 2010, 75 cases of destructive fractures were admitted in our department. According to whether the fractures interlinked with the external environment, together with the fracture sites, they were divided into 6 types: a1 type, closed diaphysis destructive fracture; a2 type, open diaphysis destructive fracture; b1 type, closed joint-involved destructive fracture; b2 type, open joint-involved destructive fracture; c1 type, closed mixed destructive fracture; c2 type, open mixed destructive fracture. Corresponding clinical treatments were conducted for each type.The new classification criterion of destructive fracture is simple and practical and thus can be used as an important guide to make reasonable treatment plans for destructive fractures.
Fracture Fixation, Internal ; Fractures, Closed ; Fractures, Comminuted ; surgery ; Fractures, Open ; surgery ; Humans

OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

ObjectiveTo measure the volume of thalamus in 1000 healthy Chinese Han nationality adults,and to analyze the relationship between thalamic volume and age,sex,weight and cerebral volume,to provide reliable data for the construction of database of Chinese adults' digital standard brain.Methods Totally 1000 healthy Chinese adults of Han nationality aged from 18 to 80 years were recruited.They were divided into 5 groups by age:18-30,31-40,41-50,51-60 and 61-80 years.Each group included 100 males and 100 females.Brain images were obtained on a 1.5 T MR,and the outline of thalami was drawn with Aquariusws software.Then the thalamic volume was calculated automatically.The volumes of left and right thalamus were compared by paired sample t-test.Thalamic volumes of the same side were compared between males and females by independent sample t-test.And thalamic volumes of different age groups were compared by one-way ANOVA.The relationships between thalamic volume and age,sex,weight and cerebral volume were analyzed respectively.ResultsThe males' standardized volumes of left and right thalamus of healthy Chinese Han nationality adults were (5776 ± 780),(5655 ± 759) mm3,and they were (5464 ±573 ),(5360 ± 542) mm3 for female.The males' thalamic volume was more than the females' on the same side ( t =2.245,2.200,P ＜ 0.01 ).The left thalamic volumes of various age groups were ( 6180 ± 534 ),(6047 ± 562 ),( 5426 ± 471 ),( 5552 ± 526 ),( 4866 ± 552 ) mm3,respectively,while the right thalamic volumes of the 5 groups were ( 6069 ± 532 ),( 5895 ± 539 ),( 5357 ± 480 ),( 5396 ± 445 ),( 4791 ± 558)mm3,respectively.There were statistically significant difference among the 5 groups( F =165.686,165.235,P ＜ 0.01 ).The left and right thalamic volume were all negatively correlated with age ( r =- 0.633,-0.645,P ＜ 0.05 ).ConclusionsWith high resolution 1.5 T MR scanner,grey matter and white matter can be depicted clearly and the outline of thalamic can be discriminated specifically.It is useful for the measurement of thalamic volume and scientific data to build Chinese adults' digital standard brain.

OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.

METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.

RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.

CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.

Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology

OBJECTIVETo study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA).

METHODRats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA.

RESULTCompared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA.

CONCLUSIONXLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

Animals ; Arthritis, Experimental ; immunology ; pathology ; prevention & control ; Cell Proliferation ; drug effects ; Colubridae ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Edema ; immunology ; pathology ; prevention & control ; Female ; Immunoglobulin G ; metabolism ; Lymphocytes ; drug effects ; pathology ; secretion ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Spleen ; pathology ; secretion ; Thymus Gland ; pathology ; Toes ; pathology

Objective To analyze the acupoint compatibility law and the application characteristics of acupuncture moxibustion treatment for depression based on complex network technology;To provide a basis and ideas for formulating prescriptions of acupuncture antidepressant therapy.Methods Related clinical literature about acupuncture for the treatment of depression was retrieved from CNKI,Wanfang Data,VIP,CBM,PubMed,Embase,Cochrane Library,and Web of Science from the establishment of the databases to 21st,Sep.2022.According to the inclusion and exclusion criteria,the literature was screened,and the clinical literature database of acupuncture in anti-depression was established.Excel 2010 was used for frequency analysis of acupoints.SPSS Modeler 18.0 was used for association rule analysis,and Gephi 0.9.5 was used for complex network analysis.Results Totally 579 articles were included,involving 172 acupoints,with a total frequency of 3 222.The results of descriptive analysis showed that the main meridians of acupoints were Governor Vessel,bladder meridian and liver meridian,and the specific acupoints such as Yuan-primary point,Shu-stream point,Back-shu point,Luo-connecting point,Eight confluence points,Front-mu point,He-sea point were frequently used.Association rules analysis showed that the combination of the highest correlation acupoints were"Yintang(GV29)-Baihui(GV20)".Through the complex network k-core analytic hierarchy process and community analysis,two core acupoint groups were finally obtained.The acupuncture angle,depth and direction were based on safety and meridian circulation direction,the tonifying and reducing method was based on syndrome differentiation,and the prescription was based on syndrome differentiation and symptomatic selection.Conclusion Acupuncture for depression has the following characteristics:the principle of acupoint selection is based on meridian selection and specific acupoints,the selection of core acupoints focuses on the treatment of spirit and the regulation of viscera,channels and functions,and the emphasis on syndrome differentiation and symptomatic acupoint selection in clinical treatment.While,it was important to pay more attention to the specificity of acupuncture and moxibustion.

OBJECTIVETo study the influence of the mutants of hepatitis B surface antigen on the cell immunity.

METHODSThe recombinant plasmids of NS2Swt, NS2S126, NS2S133, NS2S141 and NS2S145 were transfected into Chinese hamster ovary (CHO) cells and the expressed proteins were detected by means of ELISA. Following PHA-activated lymphoblasts proliferation assay and IFN-gamma, IL-2, IL-10 induction assay were done with these proteins.

RESULTSIt was identified that these proteins of HBsAg could stimulate human lymphoblasts proliferation. Besides, there were no significant difference between the mutants and the wild. It was deserved to point out that the HBsAg with T126S mutation could increase the expression of IFN-gamma in the culture medium while the HBsAg with M133T mutation induced more expression of IL-10.

CONCLUSIONThe results suggested that the cellular immune response to mutants of HBV might not be strengthened or weakened. But it should not be ignored that HBV T126S or M133T mutation may assert a potential impact on the cell immunity.

Animals ; CHO Cells ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Surface Antigens ; genetics ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Lymphocytes ; cytology ; drug effects ; metabolism ; Mutant Proteins ; genetics ; Mutation ; Plasmids ; genetics ; Recombinant Proteins ; pharmacology ; Transfection