Objective To understand the base of the neuroanatomy underlying the nociceptive syndrome occurring in cervical zygapophyseal joints,in which pain in upper limb was accompanied. Methods The capsules of the C4-T1 zygapophyseal joints of SD rats were immunostained with anti-calcitonin-gene-related peptide(CGRP) antibody,and a modified tricetry labeling was done as following:the fluorescents,fast blue(FB) and nucleus yellow(NY),were injected,respectively,into the right radial nerve and capsules of the zygapophyseal joints of neck,then, the ipsilateral dorsal root ganglia(DRG) were sectioned and the retrogradely labeled sensory neurons in the DRG were observed and photographed,finally,the sections were further immunostained with anti-CGRP antibody,eventually,the neurons in both of the photos labeled by fluorescents and anti-CGRP antibody were comparatively observed and analyzed. Results The nervers fibers containing CGRP distributed throughout the capsules of the zygapophyseal joints from C4-T1 in the rat.Most of these fibers coursed straightly and individually,and some of them anastomosed to each other to form the networks of the nerve fibers.The neurons retrograde labeled by fluorescents were observed in each of the DRG from C5-T1.These neurons shared a labeling populations of FB single(76%),NY single(16%) and FB/YN double labeling(8%) respectively.Moreover, about 40% fluorescents labeled neurons were immunoreacted to CGRP.Conclusion The peripheral processes from a population of sensory neurons that contain CGRP in the cervical DRG simultaneously innervated both the capsules of cervical zygapophyseal joints and the fore limb with the radial nerve.The present results suggest that the nociceptive syndrome of cervical zygapophyseal joints with pain in human upper limb might be the referred pain occurring in the level of the DRG.

Medical laboratories provide test data and consultation services and are engaged in ensuring test results being timely,accurate and reliable,which are satisfactory for their service recipients.A summary of the customer satisfaction theory and quality management system,and an analysis of problems found in the quality management of such laboratories,attempt to pinpoint underlying causes of service quality setbacks.Based on such studies,the quality management system is built in accordance with customer satisfaction theory and CNAS-CL02 Accreditation Criteria for the Quality and Competence of Medical Laboratories (ISO 15189:2007).Such efforts aim to continuously improve service quality and ensure customer satisfaction.

Background and purpose:The BRAF V600E mutation is a highly attractive drug target. Therefore, determining the BRAF gene mutation status of patients is essential in order to assess patients' eligibility for targeted BRAF geneinhibitor therapy. The aim of this study was to validate the utility of immunohistochemistry to rapidly obtain the BRAF gene mutation status. This study aimed to analyze the correlation of the BRAF V600E gene mutation and VE1 protein ex-pression with the clinical pathological characteristics in papillary thyroid carcinoma (PTC).Methods:The mutation status of BRAF V600E was detected by DNA sequencing. Immunohistochemistry was used to detect the expression of BRAF V600E protein in 108 cases of PTC, 54 cases of thyroid adenoma and 54 cases of normal thyroid tissue.Results:The gene mutation rate of BRAF V600E is 67.6%, and VE1 protein expression rate is 64.8% in 108 cases of PTC. The differences were statistically significant compared with thyroid adenoma and goiter (P<0.05), but have no correlation with the clinical pathological characteristics.Conclusion:BRAF V600E gene mutation and VE1 protein expression are useful biomarkers for the pathological diagnosis of PTC. High consistency was observed between the immunohistochemical staining results and the DNA sequencingresults of BRAF V600E gene mutations. Immunohistochemical technique detecting the BRAF V600E protein expression can effectively reflect indirectly BRAF V600E gene mutation status in PTC.BRAF V600E gene mutation has no contribution to the development of papillary thyroid carcinoma.

Objective To evaluate the immune response triggered by an in-house constructed hu-man metapneumovirus multi-epitope antigen ( MEA) in a mouse model .Methods Female SPF BALB/c mice at age 4-6 weeks were used in the study and divided into 7 groups.Mice in the five groups including MEA+oligodeoxynucleotides containing CpG motifs ( CpG ODN) intraperitoneal injection ( i.p.) treatment group, MEA+Alum i.p.treatment group, MEA+Alum+CpG ODN i.p.treatment group, MEA+CpG ODN intranasal (i.n.) treatment group and MEA+Alum+CpG ODN i.n.treatment group were immunized three times on days 0, 14 and 21, and those in the other experimental group were immunized intramuscularly with MEA+Quickantibody5W on days 0 and 21.A control group without treatment was set up accordingly .All mice were sacrificed two weeks after the last immunization .Antibodies including IgG , IgG1, IgG2a and IgA in serum samples were detected by ELISA .MTS assay was performed to analyze the proliferation of lympho-cytes.The cytotoxicity of cytotoxic T lymphocytes (CTL) was measured by LDH assay.Flow cytometry was used to detect T lymphocyte subsets .The cytokines secreted by T helper cells ( Th1 and Th2) were analyzed with Bio-Rad Liquid Chips.Results High titers of IgG, IgG1 and IgG2a antibodies were produced in MEA treated mice except for those in intranasal treatment groups .Serum samples from three groups including the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups were positive for IgA antibody .The highest titer of IgA antibody was detected in mice from the MEA+Alum+CpG ODN i.p.treatment group, which was 2.15×103.Compared with the control group, significantly enhanced proliferation of lymphocytes was observed in the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups (P<0.05).Enhanced cytotoxic activities of CTL were observed in mice with ip.and i.m.treatments as compared with those in control group (P<0.05).The levels of CD4+/CD8+T cells were slightly increased in mice from the MEA+CpG ODN i.p., MEA+Alum+CpG ODN i.p. and MEA+Quickantibody5W i.m.treatment groups as compared with those in control group (P<0.05).In-creased secretion of IL-2, IFN-γand Th2-type cytokines including IL-4, IL-5 and IL-10 were detected in mice from the MEA+CpG ODN i.p.treatment group.The MEA+Alum i.p.treated mice showed a slightly increased secretion of IFN-γand significantly increased secretions of IL-4, IL-5 and IL-10.Significantly in-creased secretions of IFN-γ, IL-4, IL-5 and IL-10 were detected in mice from the MEA+Alum+CpG ODN i.p.treatment group.Significantly increased secretions of IFN-γ, IL-5, IL-10 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were detected in mice from the MEA+Quickantibody5W i.m.treatment group.Conclusion MEA together with different adjuvants could stimulate high titers of specific antibodies , increase the proliferation of lymphocytes and enhance the cytotoxic activities of CTL .CpG ODN could bal-ance the Th1/Th2-mediated immune responses , and the balance could be enhanced when using CpG ODN in combination with Alum .A similar effect could be achieved by using the commercial adjuvant Quickanti -body5w.This study has paved the way for further investigation on the development of hMPV epitope vaccines and diagnostic reagents for hMPV as well as the epidemiological study of hMPV .

OBJECTIVE:To optimize the decoction technology of Zhuanggufang decoction powder. METHODS:Central com-posite design with 2 factors and 5 levels was conducted to design the test,taking water (fold) and decoction time as independent variables,the extraction amount of icariin and extraction yield overall desirability value of as dependent variables,regression equa-tion were fitted;response surface method was used to optimize the decoction technology of Zhuanggufang decoction powder,and was verified. It was compared with the traditional decoction piece effect. RESULTS:The optimized technology was decocted twice with 16-fold water,20 min each time;relative error of predicted and theoretical value by overall desirability value was 2.97%;af-ter decoction powder and piece decocted in the same conditions,the extraction amounts of icariin were 1.2343 μg/g and 1.1324μg/g,extraction yields were 23.73% and 17.84%,respectively. CONCLUSIONS:Central composite design-response surface meth-od optimizing decoction technology is simple and reliable,the optimized decoction technology is stable and feasible,and the decoc-tion capability of Zhuanggufang decoction powder is better than the traditional pieces.

Objective To investigate the therapeutic effect of Xingnaojing in treatment of hypertensive cerebral hemorrhage.Methods 90 patients diagnosed as cerebral hemorrhage patient hospitalized were randomly divided into treatment group and control group,treatment group 45 cases,control group 45 cases,including 23 males and 22 females aged 52～75 years(71 ±4).Results Treatment group:After 14 days treatment,neurological deficits had been greatly improved,the total effective was 97.3％ ;Control group:Aftenr treatment,the improvement of neurological deficit was worse,the total effective was 77.3％.Hematoma volume in the two groups did not change significantly after one week treatment,but significant changed after two weeks treatment.There was significant difference between the two groups.Conclusion Xingnaojing had significant effect in treatment of hypertensive cerebral hemorrhage.

To explore the prevalence of hepatitis B virus (HBV) in multiple myeloma (MM) patients, as well as to compare the clinical characteristics and outcome between HBV infected and non-HBV infected patients. Methods:The serology markers of HBV were detected in 363 MM patients and 11227 cases of healthy controls through chemiluminescence. HBV-DNA was measured via real-time quantitative chain reaction. Results:Sixteen out of 363 MM patients (4.4%) were HBsAg-positive, showing significant difference with healthy controls (2.4%). No statistically significant differences were observed in terms of sex, age, type of monoclonal (M) protein, International Staging System (ISS) stage, stem cell transplantation, and risk stratification between HBsAg-positive and HBsAg-negative patients. No significant effect of HBV infection was found on the OS of MM patients. HBV reactivation was observed in two HBsAg-positive MM patients who were treated with combination chemotherapy, including bortezomib and dexamethasone. The replication of HBV could be inhibited by anti-HBV drugs. Conclusion:A higher prevalence of HBV infection was revealed in MM patients. Close monitoring of HBV replication should be conducted in MM patients with HBV infection before and during the courses of chemotherapy.

Objective To investigate the effect of mice gender on the TSH receptor antibody(TRAb)titers, the levels of TT4,and the degree of thyroid hyperplasia by establishing an animal model of Graves′ disease in male and female BALB/c mice. Methods Male and female BALB/c mice were immunized with recombinant adenovirus expressing TSHRA subunit(Ad-TSHR289)to induce Graves′ disease. Animals were injected 3 times at intervals of 3 weeks. All mice were sacrificed 4 weeks after the last injection to obtain blood for measurement of TSHR antibody titers and TT4evels, and thyroid glands for histological examination. Results TRAb positive rates were 100% both in female or male mice. No significant difference was observed in titers of TRAb between them. The incidence of hyperthyroidism in female mice was higher than that in male mice, being 75.0% and 41.7% respectively. There was statistical difference in levels of TT4between females and males(P＜0.01). Mice with high TT4exihibited marked thyroid hyperplasia. Conclusion Despite TSHR antibodies were similar between female and male mice, the incidence and degree of hyperthyroidism showed sex bias in Graves′s animal model. The results indicated that it was easier to induce model in females than in males by immunizing BALB/c mice with Ad-TSHR289.

Objective To investigate the feasibility of inducing neonatal immunotolerance against Graves'disease by gene TSH receptor (TSHR) 289 and its possible mechanism.Methods Neonatal (0-24 h) female BALB/c mice were divided into intraperitoneal injection group,intramuscular injection group,model group,and normal control group.The intraperitoneal group and the intramuscular group were further divided into low-dosage,middle-dosage,high-dosage tolerance groups,and the coresponding control groups.The tolerance groups and the controls were intraperitoneally or intramuscularly pretreated with low-dosage( 1×106 particles),middle-dosage( 1 × 108particles),high-dosage( 1 × 1010 particles)of Ad-TSHR 289 or Ad-lacz respectively.6 to 7 weeks later,the normal control group received intramuscular injection with Ad-lacz; the other groups were immunized with Ad-TSHR289,three times at 3 weeks interval.10 days after the first immunization,serum TRAb was detected.4 weeks after the last immunization,serum TRAb,TT4,splenic CD4 + CD25 + Foxp3/CD4 + were tested,and the thyroid tissues were examinated histologically.Results Ten days after the first immunization,no antibody response against TSHR was detected in the two high-dose tolerance groups,but the TRAb titer in respective controls was significantly higher( P＜0.05 ).4 weeks after the last injection,in high-dose tolerance groups,only 1/10 of mice immunized by intraperitoneal or intramuscular injection elicited anti-TSHR antibody,and no mice immunized intraperitoneally had elevated serum TT4.Two of ten mice challenged intramuscularly showed slightly increased TT4 levels,but the respective controls displayed a strong antibody response( P＜0.01 ) and elevated TT4 level ( P＜0.05 ).The similar percentages of high TT4 and thyroid hyperplasia were found in all groups.Additionally,the frequencies of CD4+CD25 +Foxp3/CD4+in two high-dose tolerance groups were significantly increased as compared to those in controls( P＜0.05 ).The incidence of Graves' disease in the other groups by intraperitoneal or intranuscular injections was not statistically different from those in the corresponding control groups and the model group.Conclusions The immune tolerance against Graves'disease is induced in neonatal mice by either intraperitoneal or intramuscular pathway with specific antigen of TSHR 289,carried by adenovirus vector,and then inhibits Graves' disease in adults. Stimulation with the high-dosage antigen is liable to induce immune unresponsiveness.CD4 + CD25 + Foxp3 +T cells may play an important role in the induction and maintenance of tolerance.

Objective To compare the efficacy of Graves disease animal models induced by thyroid stimulating hormone receptor (TSHR) plasmid DNA (pcDNA3.1-TSHR) and by TSHR A subunit recombinant adenovirus(Ad-TSHR289),and to investigate the influence of duration for preparing animal model induced by Ad-TSHR289 on Graves hyperthyroidism and its related indices.Methods The plasmid group and the adenovirus group were set up respectively.The plasmid group:21 female BALB/c mice were randomly divided into model group (n =12) and control group (n =9).The model group were injected intradermally with pcDNA3.1-TSHR 50 μg,once every 3 weeks,totally 3 times.Then 4 weeks after the last immunization,the mice were euthanized to obtain blood for testing TSHR antibody (TRAb),total T4,and thyroid tissue for histological examination.The controls were injected with the same dose of pcDNA3.1 in the same way.The adenovirus group:52 female BALB/c mice were divided into 10-week model group (n =8),14-week model group (n =10) and 18-week model group (n =8),and the respective controls (n =8,n =10,n =8) were set up.All model groups were injected intramuscularly with Ad-TSHR289,three times at three weekly intervals.Then the mice were euthanized at 4,8 and 12 weeks to test TRAb,total T4 level and to observe the change of thyroid histology.The controls were treated with the same dose of Ad-lacz in the same way.Another 8 mice were scheduled to test the dynamic variation of TRAb before and after the 3 times immunization.Results In the plasmid model group,only two of 12 mice developed weak antibody responses against TSHR,and no elevated total T4 level and no hyperplasia changes of thyroid were observed.In the 10-week model group,all mice had high level TRAb [(807.65 ± 136.33)U/L,Six-eighths mice had hyperthyroidism exhibited hyperplasia changes.In the 14-week model group,the TRAb level [(650.12 ± 192.88) U/L]and the incidence of hyperthyroidism (3/10) were lower than those in 10-week group.Histologically,the degree of thyroid hyperplasia lightened to a small extent,but its positive rate did not decline.In the 18-week model group,only 2 of 8 mice displayed slightly elevated TRAb level,and no mice showed increased total T4 level.Additionally,thyroid tissues of 2 mice were mildly abnormal.Compared with the model groups at different time,the change of antibody levels of the mice for TRAb dynamic observation exhibited the similar trend.Conclusions Being good at repeatability and high incidence of hyperthyroidism,the animal model of Graves disease induced by Ad-TSHR289 is still an ideal research tool presently.The duration of model ean be maintained 18 weeks,and 10 weeks is the best period to snstain characteristic of Graves disease.