BACKGROUND: The determination of monoamine neurotransmitters and their metabolites is important in the experimental.research of Parkinson disease (PD) and experimental diagnosis of phaochromocytoma.OBJECTIVE: To develop a high performance liquid chromatography (HPLC) method for simultaneous and rapid determination of monoamine neurotransmitters and their metabolites which include epinephrine(E), norepinephrine(NE), dopamine(DA), 5'-hydroxytryptamine(5'-HT), 5-hydroxy3-indoleacetic acid(5-HIAA), homovanillic acid(HVA) and 3,4-dihydroxyphenylacetic acid(DOPAC).DESIGN: Case control observation and a randomized controlled animal experiment.SETTING: Department of Clinical Laboratory, Affiliated Hospital of Medical College, Qingdao University; Haiyang Branch, Affiliated Hospital of Medical College, Qingdao University.PARTICIPANTS: This experiment was carried out at Staff Room of Physiology, edical College, Qingdao University and Department of Clinical Laboratory, Affiliated Hospital of Medical College of Qingdao University from October to December 2003. ①Two patients with phaeochromocytoma were all confirmed by CT, MRI or pathologic section as well as physical sign. Twelve health volunteers were all university students and were excluded to suffer hypertension, endocrine system diseases and other diseases. ② Total 12 healthy female Wister rats weighed between 150 g to 180 g were chosen and randomly divided into control group and PD group with 6 rats in each.METHODS: The standard chromatograms of the standard solution of mixtures of seven substances (E, NE, DA, 5'-HT, 5-HIAA, HVA and DOPAC)were prepared and the precisions and recovery rates of the same samples were determined respectively. Urine samples of 24 hours obtained from 12 healthy volunteers and 2 patients with phaeochromocytoma were analyzed respectively by HPLC. The unilateral forebrain bundles in PD group were destroyed with 6-hydroxydopamine (6-OHDA) for monitoring release characteristics of monoamine neurotransmitters and their metabolites in healthy and marred caudate putamen nucleus striatum of normal and PD rats striatum.MAIN OUTCOME MEASURES: ①Determination of the precisions and recovery rates of monoamines. ②The contents of catecholamines in 24 hours urine of 12 health volunteers and 2 patients with phaeochromocytoma .③The release characteristics of monoamines and their metabolites in rats striatum. RESULTS: The results and analysis of catecholamines substances in 24hours urine were all obtained from 12 health volunteers and 2 patients with pheochromocytoma. There were no animals died in the experinment, and all of them were involved in the result analysis. ①There was a favorable linear relationship in the range of 2-1 000 μg/L for DA, E, NE and DOPAC, 10-1 000 μg/L for 5-HIAA, HVA and 5-HT for 30-1 000 μg/L with HPLC. The minimal detection limit of DA, E, NE and DOPAC was 2 μg/L, 5-HIAA 10 μg/L, HVA and 5-HT 30 μg/L. The coefficient of variation (CV) in the same batch was 3.0%-8.6%, and the CV in the different batchs was 3.8%-9.7%. The average rates of recovery of DA, NE, E and other 4 kinds of substances were 107.3%, 89.4%, 80.1% and 89.2%-95.7%, respectively.②The contents of CA in 24 hours urine of the 2 patients with phaeochromocytomas were higher than those of the 12 healthy volunteers. One of patient samples showed that E was mainly higher than that of control group. Other samples showed that NE was mainly higher than that of control group. Otherwise , E and NE were ten times higher than that of control group. ③The contents of DA and its metabolites such as DOPAC and HVA in intact sides of hemiparkinsonian rats had no significant changes ,while the contents of DA, DOPAC and HVA in lesion sides were significantly decreased compared with the controls (P ＜ 0.01 ).There were no significant changes in the contents of 5-HT and its metabolites 5-HIAA. ④The turnover rates of DA (DOPAC/DA and HVA/DA) in intact and lesion sides of hemiparkinsonian rats were all higher than those of the controls(P ＜ 0.05).CONCLUSION: The method we developed is suitable for the experimental research of normal and hemiparkinsonian rat models and for the diagnostic testing of phaeochromocytoma.

The total protein of Toxoplasma gondii RH strain tachyzoites was separated by two-dimensional electrophoresis (2-DE), and Western blotting was performed to find out distinct antigens. 209 spots were detected through Coomassie brilliant blue-stained small gels (7?8 cm, pH 3-10). Western blotting showed 17 specific antigen spots with sera of the experimental group and two nonspecific spots with the control sera.

Objective To evaluate the efficacy of total glucosides of paeony(TGP) in the treatment of undifferentiated spondyloarthropathies(uSpA) and investigate its mechanism.Methods One hundred and sixteen cases of uSpA were randomly assigned to 2 groups,one group was treated with TGP and diclofenac sodium,the other group with SASP and diclofenac sodium.The efficacy and side effects of the two groups were compared after 12 weeks treatment.The level of IL-6,IL-10 and TNF-? was determined at the same time.Result After 12 weeks treatment,the major clinical manifest and the indexes of disease activity were no difference of statistical significance in the two groups.The incidence of parenterally side effects was 8.6% in TGP group,and 25.9% in SASP group.There was difference at statistical significance between the two groups(? 2 =6.042,P=0.014).Before treatment,the level of IL-6,IL-10 and TNF-? in the serum of two groups were no statistical difference,while there was obviously difference after 12 weeks treatment(t= 2.672,2.483 and 2.088,P=0.009,0.014 and 0.039).Conclusion The efficacy of TGP and SASP has no significant difference in the treatment of uSpA.But the incidence of parenterally side effects in the TGP group was less than SASP group.The mechanism of efficacy of TGP may be related to inhibiting the production of IL-6,IL-10 and TNF-?.

Objective To improve the understanding of osteoarticular pain presented as rheumatic syndromes of cancers and malignant skeleton metastasis associated. Methods To analyse the clinical data of 32 patients initially manifested as osteoarticular pain. Results Forteen patients of 32 cases was diagnosed as cancer associated rheumatic syndromes, characterized by gradually deteriorated chronic recurrent vague pain. The effect of non-steroid anti-inflammatory drugs (NSAIDs) and glucocorticoid was not satisfactory. Pain subsided after removal of carcinous mass. Eighteen patients had malignant skeleton metastasis, vague pain at onset, but gradually progressed to be, severe persistent pain. NSAIDs and glucocorticoid were ineffective. Narcotic analgesics could produce short time relief. The nature and distribution of tumour were comparatively extensive. The clinical misdiagnosis rate was high. Conclusion Clinical doctors often neglect the coexistence of tumor because they are not familiar with the relation between osteoarticular pain and tumor. So patients who is over sixty and is not sensitive to commonly used analgesics and glucocorticoid with osteoarticular muscular pain should be considered the possibility of malignant tumor after excluding diffuse connective tissue disease.

Objective To detect the frequency of peripheral blood CD4+ CD25high Tr and CD4+CD25lowT cells and expression of programmed death-1 ( PD-1 ) on their surface in the patients with systemic lupus erythematosus(SLE) and their clinical signifcance is analyzed.Methods The expression of PD-1 on the CD4+ CD25highTr and CD4+CD25lowT cells was examined in patients with 33 active SLE,18 inactive SLE and 38 healthy controls(HC) by flow cytometry.Clinical manifestations and laboratory findings of SLE were collected.Patients were divided into two groups according to their disease activity.SLE disease activity index(SLEDAI) score≥10 was defined as high disease activity and ＜10 as inactivity.The percentage of CD4+ CD25highTr and CD4+CD25lowT cells and proportions of expression of PD-1 on their surface were compared between not only inactive or active SLE patients and healthy controls (HC),but also between patients with lupus nephritis and without lupus nephritis.Correlation with clincal manifestations and laboratory findings was analyzed.Results (1)The proportions of CD4+CD25highTr and CD4+CD25low T cells were significantly decreased in active and inactive SLE patients as compared with HC.(2)The percentage of CD4+ CD25lowPD-1 +Tr and in active and inactive SLE patients were higher than HC.The proportions of CD4+CD25lowPD-1+ T cells were significantly increased in HC and inactive SLE patients as compared with inactive SLE patients.(3) The proportions of CD4+CD25highPD-1+Tr in SLE patients with nephritis were higher than those in patients without nephritis.But the percentage of CD4+ CD25 lowPD-1 + T cells was significantly decreased in SLE patients with nephritis as compared those without nephritis.(4)A positive correlation was observed for proportions of CD4+CD25highPD-1 + T cells with SLEDAI score.Proportions of CD4+ CD25high Tr,CD4+ CD25low PD-1 + T cells was inversely correlated with SLEDAI score.Conclusion The aberrations of proportions of CD4+CD25highpD-1+,CD4+CD25lowPD-1+,CD4+CD25high and CD4+CD25low T cells were observed in patients with SLE.The abnormality of PD-1 and PD-L1 pathway may play an important role in the function and number of Tr.

Objective:To study the clinical effect of Maizhiling tablets combined with salmon calcitonin in the treatment of elderly femoral neck fracture after artificial femoral head replacement and the influencing factors of hip joint function 6 months after operation.Methods:Prospective research methods were used in this article.A total of 84 elderly patients with femoral neck fracture who underwent artificial femoral head replacement in the Affiliated Hospital of North China University of Science and Technology from January 2017 to June 2019 were selected and randomly divided into the treatment group (41 cases) and the control group (43 cases) by the random number table method.The treatment group was treated with Maizhiling tablet and salmon calcitonin nasal spray plus basic calcium; the control group was treated with basic calcium.Independent sample t test was used to compare the visual analogue score (VAS) on the 5th day after operation, the time of complete disappearance of congestion in thigh, and the time of first ambulation after operation between the two groups.The bone mineral density (BMD) of contralateral femoral neck and Harris score of hip joint function were compared between the two groups at 14 days, 3 months and 6 months after operation by repeated measurement variance analysis.Multivariate Logistic regression was used to analyze the influencing factors of hip joint function at 6 months after operation.Results:VAS score on the 5th day after surgery: (6.34±1.54) points in the treatment group and (7.02±1.50) points in the control group.Complete regression time of thigh congestion: (12.12±2.27) d in the treatment group and (13.88±2.58) d in the control group.The first postoperative activity time was (5.93±1.52) d in the treatment group and (7.84±1.05) d in the control group, and the differences of the above indexes between the two groups were statistically significant ( t value was 2.051, 3.313, 6.673, respectively, all P<0.05). The BMD values of the contralateral femoral neck in the treatment group were (0.543±0.123), (0.561±0.119), (0.606±0.105) g/cm 2 at 14 days, 3 months and 6 months after operation, and (0.530±0.092), (0.517±0.089), (0.526±0.090) g/cm 2 in the control group.The results of variance analysis of repeated measurements showed that Fintra-group=55.726, P<0.01; Finter-group=4.206, P<0.05; Finteraction=57.654, P<0.01.There were significant differences between 3 months, 6 months and 14 days after operation in the treatment group (all P<0.01), and between 3 months and 6 months after operation ( P<0.01). In the control group, there was significant difference between 14 days and 3 months ( P<0.01), and between 3 months and 6 months ( P<0.01); 6 months after operation, the difference was statistically significant ( P<0.01). The Harris scores of hip joint in the treatment group were (82.12±8.18), (85.49±6.61), (87.10±6.57) points and (78.91±5.75), (81.44±6.42), (83.67±6.97) points in the control group.The results of repeated measurement ANOVA showed that Fintra-group=64.526, P<0.01; Finter-group=6.529, P<0.05; Finteraction=0.501, P>0.05.There were significant differences between 3 months, 6 months and 14 days after operation (all P<0.01). There were significant differences between 3 months and 6 months after operation ( P<0.05). There were significant differences between the groups 14 days, 3 months and 6 months after operation ( P<0.05 or P<0.01). Multivariate logistic regression analysis showed that age( OR=1.318, 95% CI1.002-1.732, P=0.048) and treatment method ( OR=29.168, 95% CI 1.030-824.623, P=0.048) were risk factors for hip function on the replacement side, and Harris score of hip function 14 days after surgery ( OR=0.624, 95% CI 0.447-0.873, P=0.006) were protective factors for hip function 6 months after surgery. Conclusion:Maizhiling tablets and salmon calcitonin nasal spray have good effects on removing blood stasis, relieving pain, effectively improving bone mineral density and promoting the recovery of joint function after artificial femoral head replacement for senile femoral neck fracture.The younger age of patients, postoperative treatment with Maizhiling tablets combined with salmon calcitonin and good recovery of hip function in 14 days after operation are the protective factors of good hip function in 6 months after operation.

Objective To investigate the effects and toxicity of pirarubicin and chemotherapy regimen in treatment of malignant lymphoma in the elderly. Methods 50 cases(over 60 years of age) with non Hodgkin's lymphoma collected in Lishui Central Hospital of Zhejiang province from January 2011 to January 2013 were randomly divided into control group(n=25)and experimental group(n=25). Experimental group were received CTOP chemotherapy, control group were received the standard CHOP chemotherapy, vomiting, stomach, blood pressure, blood glucose control and other appropriate symptomatic treatment were adopted according to the speciifc circumstances of each patient in both two groups. Regular inspection were given to all patients, and bone marrow suppression, treatment of lymphoma, complications and other indicators were observed. Results Complete remission in experimental group after treatment was 13 cases, accounted for 52%within group, partial remission were 9 cases, accounted for 36%. The total effective number was 22, and the rate was 88%. Complete remission in control group after treatment was 8 cases, accounted for 32%within group, partial remission were 5 cases, accounted for 20%. The total effective number was 13 cases, and the rate was 52%. Bone marrow suppression in experimental group was signiifcantly less than control group (P<0.05). Conclusion Elderly patients with malignant lymphoma in chemotherapy plus pirarubicin can improve the therapeutic effect, reduce bone marrow suppression and complications.

AIM: To observe the effects of H_2O_2 on apoptosis of skeletal muscle satellite cells (SMSC)and mitochondrial membrane potential (MMP), and protective effect of erythropoietin(EPO). METHODS: SMSC in vitro were divided into three groups: H_2O_2 group, H_2O_2+EPO group and control. Apoptosis rate and the means were obverted by monofluorescence flow cytometry. The morphological change of apoptosis cells were observed under fluorescence microscopy after Hoechst 33258 staining. RESULTS: The cells in H_2O_2 group show the highest apoptosis rate (22.13±1.79)%. In H_2O_2+EPO group, apoptosis rate were (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% according to the EPO treated levels (10, 20 or 40 kU/L), respectively. MMP level in H_2O_2 group was the lowest 9.70±0.09. MMP levels in H_2O_2+EPO group were 12.67±0.32, 27.90±0.66, 44.53±0.93, respectively according to the EPO treated levels (10, 20 or 40 kU/L). In control group, apoptosis rate was 1.93±0.57 and MMP was 51.37±0.64. In H_2O_2 group and H_2O_2+ low dosage EPO group, Hoechst 33258 staining showed obvious apoptosis. CONCLUSION: EPO inhibits the apoptosis induced by H_2O_2 and stabilizes the MMP, which is related to the dosage of EPO.

Objective To investigate the association between a few single nucleotide polymorphisms (SNPs)in Fc receptor like 3(FcRL3)and rheumatoid arthritis(RA)in Han nationality of Anhui patients.Methods One hundred and forty RA patients along with one hundred and eighty-seven healthy controls were included in the study.SNPs of FcRL3-1(rs0158440),FcRL3-2(rs2225828),FcRL3-3(rs7528684).FcRL34(rs11284799),FcRL3-5(rs945635),FcRL3-6(rs3761959),FcRL3-7(rs2210913),FcRL3-8(rs2282284)and FcRL3-9(rs2282283)in the FcRL3 gene were genotyped by MALDI-TOF technology.Haplotypes were estimated using PHASE v 2.1 software.Results The frequency of FcRL3-3-1 69C alleles in RA patients was significantly increased compared with healthy controls(X~2=7.348,P=0.007,OR=1.558,CI:1.130～2.150).The decreased compared with healthy controls.ConclusionFcRL3 may be associated with RA susceptibility in Anhui Han population.

Objective To obtain genes encoding the novel molecules for diagnosis of schistosomiasis.Methods Juvenile S.japonicum cDNA library was immunoscreened to obtain positive clones.By DNA sequencing and sequence analysis,the target gene was amplified by PCR and subcloned into prokaryotic plasmid pET28a.The recombinant plasmid was transformed into E.coli BL21 followed by expression of the protein induced by IPTG.The protein was identified by Western blotting.Results 34 positive clones were obtained,24 of which were chosen to be sequenced,13 of which were Sj22600 gene.The protein were recognized with sera of infected rabbits and patients with acute and chronic schistosomiasis by Western blotting.Conclusion The gene coding for Sj22600 membrane protein was screened with high frequency in the cDNA library.The E.coli BL21 transformed with the recombinant plasmid can express the fusion protein,which shows immunoactivity.