Objective To determine whether p38 mitogen-activated protein kinase (p38MAPK) signaling pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.Methods Two hundred and twenty-five male Kunming mice weighing 30-40 g were randomly divided into 4 groups:group control ( group C,n =55 ) ;group fractalkine (group F,n =60); group anti-CX3CR1 + fractalkine (group CF,n =55) and group SB203580 (p38MAPK inhibitor) + fractalkine (group SF,n =55).Fractalkine 100 ng was injected into cerebral lateral ventricle (i.c.v.) in groups F,CF and SF.Anti-CX3CR1 1 μg and SB203580 1 μg were injected i.c.v.at 1 h before fractalkine injection in groups CF and SF respectively.Paw withdrawal latency to a thermal nociceptive stimulus (PWL) was measured at 30 min before the drugs were injected into cerebral lateral ventricle and 30,60,120 and 240 min after fractalkine injection.Five animals were sacrificed after PWL measurement at each time point and their brains were removed for determination of phosphorylated p38MAPK protein expression (by Western blot analysis).Five animals were sacrificed at 30 min before the drugs were injected into cerebral lateral ventricle and 6,12 and 24 h after fractalkine injection for determination of IL-1β and TNF-α contents in the brain (by ELISA) in all the 4 groups.In group F 5 animals were sacrificed at 4 h after fractalkine injection for determination of action of fractalkine on microglia or astrocyte (by immunofluorescence).Results Fractalkine i.c.v.injection significantly reduced PWL and increased phosphorylated 38MAPK,IL-1β and TNF-α levels in group F as compared with group C.Pretreatment with anti-CX3CR1 or SB203580 significantly decreased fractalkine-induced hyperalgesia and phosphorylated-p38MAPK,IL-1β and TNF-α levels in groups CF and SF as compared with group F.Fractalkine was localized at microglia.Conclusion p38MAPK signal transduction pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.

Objective To observe the perioperative management of cardiac surgery and extracorporeal circulation method in patients with glucose-6-phosphate dehydrogenase deficiency(G6PD).Methods Ten patients with G6PD deficiency underwent uneventful cardiac surgery procedures between January 2005 and December 2010.Twenty patients who had non-G6PD deficiency were as a control group,the selected conditions were the same gender,age,body mass,the risk of heart disease surgery.The preoperative management in patients with G6PD deficiency mainly focused on avoiding the drugs implicated in haemolysis,reducing the surgical stress,using moderate hypothermia extracorporeal circulation and enhancing blood conservation.Observed indicators included the assisted ventilation time,urine volume,the drainage volume of chest tube,the amount transfusion of red blood cells and plasma,the level of hemoglobin and serum total bilirubin in the 2nd day after surgery,ICU stay.Results Compared with the control group,patients with G6PD deficiency had no significant difference in duration of ventilation after the operation,drainage,urine,Hgb,bilirubin levels,and blood transfusion[(9.3 ± 4.5)h vs (8.6 ± 5.7)h,(2100 ±670)ml vs (1950 ± 490) ml,(253 ± 146)ml vs (260 ± 120)ml,(1.3 ± 1.0)U vs (1.8 ± 1.2)U,(96 ± 25)g/L vs (99 ± 12)g/L,and (24 ± 8)μmol/L vs (27 ± 1 l)μmol/L,t =0.978,2.032,1.257,0.891,2.182,2.271,and 1.329,all P ＞ 0.05].The duration of ICU discharge was significantly longer in the glucose-6-phosphate dehydrogenase deficient group[ (2.6 ± 0.6)d vs (1.8 ± 1.5)d,t =2.704,P ＜ 0.05].Conclusions Cardiac surgery can be performed safely in patients with G6PD deficiency with enhanced perioperative management.

Objective　To explore the expression of Fas ligan d (FasL) protein in human non-small cell lung cancer (NSCLC) and its clinic al significance. Methods　Expression of FasL protein was detecte d by immunohistochemical method in 32 resected tumors of NSCLC. Results 　FasL protein was detected in all of these 32 resected tumors with variant pos itive expression levels ranging from 3.0% to 98.7% ［mean (62.0±32.0)%］. T here wa s significant difference between the tumors of different pathological types no s ignificant difference was found between the tumors of different differentiation nor among the tumors of different pTNM stages (P>0.05). Conclusion　 The results indicated that NSCLC can counterattack the immune cells of t he body and may be the molecular basis for the easily metastasis of adenocarcino ma to in the early stage.

Objective: To analyse the relating factors that influence the prognosis of chronic subdural hematoma(CSDH) after drilling drainage and to explore the bilateral bleeding risk factors of CSDH. Methods: Four hundred and forty patients with CSDH who underwent drilling drainage were retrospectively analysed. Clinical data, including medical history, signs, blood coagulation function and imaging examination were collected. Spearman correlation analysis and ordinal Logistic regression analysis were performed to assess the relationships between various variables and the prognosis, and univariate analysis and multivariate Logistic regression analysis were performed to explore the risk factors that lead to the occurrence of bilateral bleeding. Results: Spearman correlation analysis indicated that age, medication history of anticoagulant drugs, bilateral hematoma or not, the thickness of the hematoma and volume of intracranial gas on the first post-surgery day and preoperative Markwalder grade were associated with postoperative Markwalder grade (P<0.05). Ordinal Logistic regression analysis showed that medication history of anticoagulant drugs and preoperative Markwalder grade were the independent risk factors (P=0.028, P=0.000). Univariate analysis of the bilateral bleeding indicated that age, medication history of anticoagulant drugs and thrombin time were statistically different between unilateral and bilateral CSDH (P<0.05). Multivariate Logistic regression analysis revealed that thrombin time was the independent risk factor (OR=1.147, 95%CI 1.005-1.309, P=0.042). Conclusion: Age and medication history of anticoagulant drugs were closely related to the bilateral chronic subdural hematoma. The higher the preoperative Markwalder grade is, the higher the postoperative Markwalder grade is. The prognosis of patients without anticoagulant drugs is better than those users.

The T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is an inhibitory receptor mainly expressed on active T-cells, or natural killer cells (NK cells) that activate negative stimulus signals in immune cells by combining with multiple ligands on the surface of target cells including tumor cells and infected cells. TIGIT plays an important regulatory role in the immune pathogenesis of tumors, viral infections and various autoimmune diseases by inhibiting the over activation of cells and the over secretion of proinflammatory cytokines. Recent researches show that TIGIT is highly expressed in T cells and NK cells of cancer patients, and is related to disease progression and poor clinical prognosis. Researchers try to enhance the activity of T cells or NK cells by blocking the binding of TIGIT and its ligand for therapeutic intervention. At present, there have been many reports about the use of anti-TIGIT monoclonal antibody treatment in different mouse tumor models leading to tumor regression, TIGIT has received extensive attention in cancer immunotherapy as a promising target for next generation cancer immunotherapy. Several clinical trials are currently evaluating the efficacy of anti-TIGIT monoclonal antibodies (mAbs) in patients with several cancers. The most advanced candidate, tiragolumab, has exhibited remarkable efficacy in programmed cell death ligand 1 (PD-L1)-positive non-small cell lung carcinoma (NSCLC) patients in phase Ⅱ clinical trials, in combination with PD-L1 blockade. However, the specific mechanism of TIGIT blockade remains to be fully elucidated.

OBJECTIVETo discuss the anti-tumor effect of Xihuang pill on tumor-bearing rats and its effect on the immune clearance function of tumor-bearing organisms.

METHODWalker256 tumor cells were adopted to establish the tumor-bearing rat model. The rats were randomly divided into five groups: the normal control group, the model control group, the lentinan group and Xihuang pill low dose, middle dose and high dose groups, with 10 rats in each group, and continuously treated and given drugs for 14 d after modeling. Blood and tumors were collected from abdominal aorta to calculate the tumor inhibition rate. The content of CD3+, CD4+, CD8+ T cells and adhesion molecule B7-1 (CD80) in peripheral blood were detected by flow cytometry (FCM). The expressions of IL-2 and IFN-gamma in were determined by ELISA.

RESULTThe tumor inhibition rate of the Xihuang pill high dose group was 33. 1 percent. Compared with the model group, the Xihuang pill large dose group showed significantly low IL-2, IFN-gamma, CD3+, CD4+, B7-1 in peripheral blood, with statistical significance in their differences (P < 0.05).

CONCLUSIONXihuang pill could show its anti-tumor effect by enhancing the immune clearance function and increasing IL-2, IFN-gamma, CD3+ T, CD4+ T, B7-1 in peripheral blood.

Animals ; Antineoplastic Agents ; administration & dosage ; B7-1 Antigen ; genetics ; immunology ; Breast Neoplasms ; drug therapy ; genetics ; immunology ; CD4-Positive T-Lymphocytes ; drug effects ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flow Cytometry ; Humans ; Immune System ; drug effects ; Immunologic Factors ; administration & dosage ; Interferon-gamma ; genetics ; immunology ; Interleukin-2 ; genetics ; immunology ; Rats ; Rats, Wistar ; Tumor Burden ; drug effects

OBJECTIVE: To establish a 15-plex rapid STR multiplex amplification system. METHODS: Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives. RESULTS: Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure. CONCLUSION The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Alleles ; Chromosome Mapping ; DNA/genetics* ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Humans ; Microsatellite Repeats/genetics* ; Polymerase Chain Reaction/methods* ; Racial Groups/genetics* ; Sensitivity and Specificity ; Tandem Repeat Sequences

Objective:To investigate polymorphism of human leukocyte antigen (HLA)-DRB1 and -DQB1 genes in Bai ethnic group in Dali,Yunnan province.Methods:Polymerase chain reaction-sequence specific primers (PCR-SSP) were used to determine HLA-DRB1 and -DQB1 alleles in 124 unrelated healthy Bai ethnic individuals living in Eryuan County of the Dali Bai autonomous prefecture,Yunnan province.Results:Among all the 21 DRB1 alleles and 15 DQB1 alleles were identified,the predominant alleles were DRB1*1202(26.61%),DRB1*0901(13.89%) and DRB1*0803(9.92%) on DRB1 locus and DQB1*0301(31.45%),DQB1*0601(10.08%),DQB1*0401(8.06%)and DQB1*0502(8.06%)on DQB1 locus.The most common haplotypes were DRB1*1202-DQB1*0301(20.08%)and DRB1*0803-DQB1*0601(7.19%).Conclusion:The phylogenetic tree constructed according to the HLA-DRB1,-DQB1 allele frequencies of Bais with those of other 10 populations suggests that the Bai ethnic group belongs to the southern group of China,but it keeps genetic distance from others and the HLA genes exhibits a unique profile.This study would provide HLA polymorphism information of Bai for the future investigation on the disease related to the genetic polymorphism.

Adolescent ; Adult ; Aged ; Biopsy ; Delayed Graft Function ; diagnosis ; pathology ; Female ; Graft Rejection ; diagnosis ; pathology ; Humans ; Kidney ; pathology ; physiopathology ; Kidney Function Tests ; Kidney Transplantation ; pathology ; Male ; Middle Aged ; Young Adult