Objective To validate the clinical performance of a domestic fibrinogen reagent by freezing method(Clauss method) on the coagulometer Japan Sysmex CA1500.Methods The domestic fibrinogen reagent as the reagent A and the Germany Siemens Dade thrombin reagent as the reagent D,the Clauss method was adopted to measure the within-run precision and between-run preci-sion in two levels of quality control respectively.The reference value range was verified by the reagent A in 165 cases of normal clin-ical samples.The fibrinogen detection results in 200 cases of clinical samples were compared between by the reagent A and the rea-gent D.The significance test and the equivalence test were performed.Results The within-run precision CV of the reagent A and D in two levels of quality control were 4.28%,6.98% and 3.45%,5.22% respectively,the between-run precision CV of the reagent A and D in two levels of quality control were 6.23%,10.34% and 6.20%,9.89% respectively,the differences had no statistical significance(P >0.05).The reference value range of the reagent A was 2.08 -3.92 g/L.The fibrinogen detection results of the clinical samples by the reagent A and D had the statistically significant differences (P =0.025).But the 90% bilateral confidence in-terval(90%CI :-0.09,0.15)of the difference in the mean detection results by these two reagents located in the equivalent interval (-0.27,0.27).Conclusion The domestic fibrinogen reagent for Clauss method has reliable detection results and is suitable for the coagulometer Japan Sysmex CA1500,which is equivalent to the clinical application of Germany Siemens Dade thrombin reagent.

Objective To study the effects of resin micro-leakage of different angles of cavity margins with composite resin filling.Methods Thirty extracted human molar with occlusal surfaces preparing hole into 3mm × 3mm × 3mm.Specimens were randomly divided into 3 groups,1 mm slopes of cavity margins were designed respectively,and three kinds of angles of cavity margins(45°,70°,90°)were filled on the ocelusive surfaces,then using Prime&Bond NT and Composite resin(ChrismaA2)to repair the holes directly.After 500 thermal cycling,all specimens were immersed in Indian ink for one week and the penetration depth was examined under stereo microscope.Results Cavity margin angles of 70°was better than those of 90° and 45°,the results was statistically significant(t=3.530,4.179,all P＜0.05).Conclusion To reduce the occurrence of the resin micro-leakage,cavity margin angles should be designed as short slant with an angle of 70°or so.

ObjectiveTo investigate the relationship between polymorphism in the PITX3 gene and hereditary susceptibility of Parkinson's disease (PD). MethodsThree PITX3 single nucleotide polymorphisms ( SNPs ),including rs2281983,rs4919621 and rs3758549 were examined in 509 late-onset PD patients ( LOPD ),290 early-onset PD(EOPD) and 494 healthy controls.Genotyping was carried out in all subjects using a ligase detection reaction( LDR).ResultsAllele and genotype frequencies did not differ between the 799 PD patients and 494 controls ( P values of genotype were 0.494,0.343,0.951 ; P values of allele were 0.369,0.297,0.823 ),between 509 LOPD patients and 494 controls ( P values of genotype were 0.522,0.350,0.630 ; P values of allele were 0.413,0.328,0.571 ),between 290 EOPD patients and 494 controls ( P values of genotype were 0.499,0.492,0.552; P values of allele were 0.321,0.301,0.931 ),and between 509 LOPD and 290 EOPD patients ( P values of genotype were 0.577,0.710,0.127 ; P values of allele were 0.346,0.472,0.077 ) for all three SNPs (rs2281983,rs4919621 and rs3758549).There were no association petween the three PITX3 SNPs and PD.ConclusionThree PITX3 SNPs do not contribute to the risk of developing PD in Chinese population.

Objective To observe the relation between onset age and the motor complications involved Parkinaon' s disease (PD).Methods The detailed clinical information of 195 patients with idiopathic PD and good response to L-dopa were recorded and followed up.The data were calculated with SPSS statistic software.Results Although the time interval between the onset of the disease and the use of L-dopa was significantly longer in the 57 early-onset patients as compared to the 138 later-onset ones ((3.88±3.33) years vs (2.36±2.21) years, t = 3.142, P = 0.002), the time interval between the use of L-dopa and the occurrence of motor complications was not significantly longer in the early-onset group ((3.81±2.06) years vs (4.24±2.00) years, t = -0.888, P = 0.378).There was no difference in the constituent ratio of non-L-dopa use from onset between early-onset and later-onset PD groups (28.07% (16/57) vs 27.54% (38/138), χ2 = 0.006, P = 0.940).There was also no difference in the dosage of daily L-dopa use when motor complications occurred between early-onset and later-onset PD groups ((601.8± 296.7) mgvs (655.6±192.5) mg, t = -0.912, P=0.365).Seven-tenths of the patients with an onset age younger than 40 years old carried the risk of motor complications after using L-dopa for 5-years and those older than 70 years had the risk at a rate of 1/10.Conclusions Delaying the of use of L-dopa may not necessarily delay the onset of motor complications.The high incidence of motor complications among younger patients may not be related with drug dosage and the type of drug firstly chosen.Younger onset age does inerease the ineidence of motor complicatious.

Objective To investigate the expressions of matrix metalloproteinases(MMPs) in Kashin-Beck disease(KBD) cartilage as well as in a KBD rat model of T-2 toxin poisoning under selenium deficient conditions, and to investigate the effect of T-2 toxin on MMP-13 expression in human chondrocytes in vitro in order to determine a possible mechanism underlying KBD. Methods Samples of articular cartilage were divided into 2 groups:controls(samples from 5 normal children, traffic accident or operation), and KBD(samples from 5 children with KBD, auctopsy). Thirty-two Sprague-Dawley rats were divided into two groups by body weight using random number table: normal diet group(n = 16) and selenium-deficient diet group(n=16). The selenium level in normal diet was 101.500μg/kg, and in selenium-deficient diet was 1.118μg/kg. Rats were fed for 4 weeks with selenium-deficient or normal diet, respectively. After successful build up of the low selenium rat model, normal diet group was then subdivided into 2 sub-groups: normal group(n = 8) and normal diet plus low T-2 toxin group(n = 8);and selenium-deficient diet group was also subdivided into 2 sub-groups: selenium-deficient group ( n = 8 ) and selenium-deficient diet plus T-2 toxin group ( n = 8 ) . T-2 toxin of 100 μg·kg-1·d-1 was administered by intragastric administration for 30 days. Then the rats were sacrificed, and their knee joints were processed for histopathological evaluation. MMP-1 and MMP-13 locations in cartilages were performed by inmmunohistochemistry. Human chondrocytes C28/I2 were cultured in vitro. The experiment was divided into 4 groups: empty vector plasmid group, MMP-13 promoter plasmid group, MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40 μg/L T-2 toxin group. MMP-13-luciferase reporter plasmid and vector plasmid were transiently transfected into C28/I2 cells for 24 hours, and then treated with 20 - 40 μg/L T-2 toxin for 24 hours. Transactivation of human MMP-13 promoter was analyzed using luciferase reporter constructs containing sequences spanning-1602 to+20 bp in C28/I2 chondrocytes. Results The percentages of chondrocytes staining for MMP-1 in the superficial and middle zones of KBD samples [(29.73 ± 10.12)%, (28.27 ± 0.91)%] were significantly higher than those of controls[(2.47 ± 0.11)%, (0.00 ± 0.00)%, all P < 0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of KBD samples [(13.21 ± 4.32)%, (41.85 ± 6.32)%] were significantly higher than those of controls[(5.72 ± 0.31)%, (0.00 ± 0.00)%, all P<0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of rats fed with selenium-deficient diet plus T-2 toxin group[(13.21 ± 4.32)%, (61.85 ± 8.68)%] were significantly higher than those of the normal and selenium-deficient groups[(2.43 ± 0.22)%, (5.89 ± 0.69)%, (3.03 ± 0.29)%, (25.99 ± 0.57)%, all P < 0.05]. Moreover, T-2 toxin activated the MMP-13 promoter detected with luciferase reporter assays in C28/I2 cells. The luciferase activities in MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40μg/L T-2 toxin group(0.082 78 ± 0.008 40, 0.103 35 ± 0.013 19) were significantly higher than those in empty vector plasmid group and MMP-13 promoter plasmid group(0.024 19 ± 0.000 96, 0.040 32 ± 0.003 56, all P < 0.05). Conclusions These data suggest that T-2 toxin induces cartilage matrix degradation through up-regulation of MMP-13 promoter expression. Increased MMPs staining intensity in KBD cartilage and the rat KBD model of T-2 toxin poisoning under selenium deficient conditions suggest that matrix degradation appear to be driven by MMPs activity.

Objective To investigate the expressions of interleukin-1β(IL-1β), interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in cartilage of children with Kashin-Beck disease(KBD) in order to provide a possible mechanism of the disease. Methods Articular cartilage tissues of 5 KBD children(KBD group) were selected from KBD children autopsy samples keeping in Institute of Endemic Diseases, Medical School of Xi’an Jiaotong University; articular cartilage tissues of 5 normal children ( control group ) were selected from non-KBD areas of Shaanxi Province, three cases were from accident death children, two cases were the samples of congential malformation of six finger. Expressions of IL-1β, IL-6 and TNF-α in the cartilage were detected using immunohistochemistry; the cells of articular cartilage were divided into three areas (superficial zone, middle zone and deep zone) to analyze the expressions of IL-1β, IL-6 and TNF-α. Results The expressions of IL-1β in superficial zone , middle zone and deep zone of articular cartilage of KBD group (63.50 ± 7.19, 54.75 ± 5.50, 66.20 ± 9.91) were significantly higher than those of control group(5.75 ± 1.26, 0.00 ± 0.00, 0.00 ± 0.00, all P<0.05). The expression of IL-6 in superficial zone of articular cartilage in KBD group(55.25 ± 6.24) was significantly higher than that of control group(0.00 ± 0.00, P<0.05). The expressions of TNF-αin all zone of articular cartilage of KBD group(33.25 ± 6.50, 3.75 ± 0.96, 29.80 ± 1.92) were significantly higher than those of control group (3.74 ± 0.82, 0.00 ± 0.00, 0.00 ± 0.00, all P < 0.05). Conclusion The levels of IL-1β, IL-6 and TNF-α are up-regulated in articular cartilage of KBD children, suggesting that cytokines may play an important role in matrix degradation in KBD children cartilage.

BACKGROUND: The transplantation of microencapsulated cell is becoming a hotspot modality in the therapy of Parkinson disease (PD). The application of Alginate-polysysine-alginate (APA) is currently limited due to fragility and pericystic fibrosis although it has been used in clinic. In this study, the native Alginate-chitosan-alginate(ACA)microencapsulated pheochromocytoma cells (PC12 cells) are transplanted into the region of corpus striatum in the injured side of the brain of the PD rat model, the functional recovery of rotational behavior and pathological changes are also observed in the control, sham and treated groups.OBJECTIVE: To observe whether the transplantation of ACA microencapsulated PC12 cells into the brain can improve the rotational behavior in the rat model of PD.DESIGN: Randomized controlled experiment.SETTING: Dalian Research Institute of Physiochemistry, Chinese Academy of Sciences.MATERIALS: Totally 40 adult male Wistar rats with body mass of(220±10) g, ACA microcapsule and PC12 cells were used in this study.METHODS: The experiment was carried out in the animal experimental laboratory of Second Hospital, Jilin University and Dalian Research Institute of Physicochemistry, Chinese Academy of Sciences between May and December 2002. Native ACA were used to microencapsulate the PC12cells. These rats were randomly divided into the following three groups,treated group (10 rats received microencapsulated PC12 cell transplantation), control group (7 rats received unencapsulated PC12 cell transplantation) and sham group (6 rats received empty microencapsule transplantation). The transplantation site was the region of corpus striatum in the injured side of brain. The difference of rotational behavior included by apomorphine was compared before and after the transplantation in these rats,the morphological changes of the transplanted microcapsules and activity of the microencapsulated cells were also detected.MAIN OUTCOME MEASURES: ①Rotational behavior of the rats before and after transplantation. ②Pathological change in the regions of substantia nigra and corpus striatum. ③ The integrality of retrieved microencapsule and the bioactivity of retrieved PC12 cells.RESULTS: ① At the 4th week of transplantation, rotational behavior was significantly decreased in the encapsulated PC12 cells treated group compared with that of the groups received empty microencapsules transplantation [(6.9±2.8),(10.5±1.6) r/min, P ＜ 0.05].Tbis behavioral improvement could last at least three months. Although the unencapsulated PC12 cells also can improve the rotational behavior compared with before transplantation[(5.6±l.1 ), (9.5±1.5) r/min, P ＜ 0.05], which only lasted two months and fetal tumor formed in the skull of some rats. There was no significant difference in rotational behavior of the rats before and after transplantation in the empty microencapsule transplantation group. PC12 cells of retrieved microencapsulate grew well after re-culture, and have bioactivity.CONCLUSION: Transplantation of ACA microencapsulated PC12 cells into the brain can improve can improve the rotational behavior of rat PD model induced by apomorphine. ACA microcapsule can both isolate the host's immune system effectively and prevent the formation of tumor, and have a promising application in clinic.

Objective To investigate the death of chondrocytes in rats which feed with T-2 toxin under selenium (Se) deficient conditions.Methods Thirty two healthy male SD rats were divided into two groups by weight which were normal diet group and Se deficiency diet group,16 rats in each group.Rats in normal diet group were fed with Se 101.5 μg/kg diet,and rats in Se deficiency diet group were fed with Se 1.1 μg/kg diet for 30 d.Normal diet group was divided into control group and T-2 toxin group,and Se deliciency diet group was randomly divided into Se-deficiency group and Se-deficiency plus T-2 toxin group,8 rats in each group.After that,rats in T-2 toxin and Se-deficiency plus T-2 toxin groups were administrated intragastrically with T-2 toxin (100 μg/kg) everyday for 30 d.Rats were put to death,the left knee was taken and stained with hematoxylin-eosin and SafraninFast green,pathological changes of rat's knee joint cartilage were observed under light microscopy,expression levels of active caspase-3 and receptor interacting protein 3 (RIP3) in rat's articular cartilage cells were determined via the immunohistochemical method.The apoptosis was also detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL).Results Red ghost outlines of chondrocyte and multiple chondral cell clusters surrounded the non-cell areas in deep zone of articular cartilage of knee joint stained with hematoxylineosin were seen in Se-deficiency plus T-2 toxin group under light microscope.In the superficial zone of cartilage,the positive percent of TUNEL and active caspase-3 in Se-deficiency plus T-2 toxin group was higher than those of control group,Se-deficiency group and T-2 toxin group [(7.47-± 0.34)％ vs (4.68 ± 0.54)％,(2.67-± 0.64)％,(2.56 ±0.54)％;(4.75 ± 0.67)％ vs (1.24 ± 0.25)％,(0.00 ± 0.00)％,(0.00 ± 0.00)％,P ＜ 0.05].In the middle zone of cartilage,the positive percent of TUNEL,active caspase-3 and RIP3 in Se-deficiency plus T-2 toxin group was significantly higher than those of control group,T-2 toxin group and Se-deficiency group [(72.06 ± 6.15)％ vs (16.10 ± 3.00)％,(19.57 ± 3.49)％,(19.33 ± 5.19)％;(51.13 ± 4.18)％ vs (10.97-± 3.01)％,(15.36 ± 4.37)％,(15.23 ± 3.13)％;(25.91 ± 13.39)％ vs (1.59 ± 1.14)％,(4.32 ± 2.91)％,(7.50 ± 5.00)％,P ＜ 0.05].The positive percents of TUNEL,active caspase-3 and RIP3 were not significantly different in the deep zone (P ＞ 0.05).Conclusion The death of the middle zone in the rat cartilage induced by T-2 toxin under selenium deficient conditions isapoptosis and necroptosis.

The researches on quality evaluation, chemical composition, pharmacological actions and plantation of Amomum villosum in Xishuangbanna was reviewed. The further research point were proposed.
Amomum ; chemistry ; growth & development ; Analgesics ; pharmacology ; Animals ; China ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Oils, Volatile ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; growth & development

Objective:To investigate the characteristics and associated factors of depression in elderly patients with Parkinson's disease(PD).Methods:A total of 1138 elderly PD patients from Xuanwu Hospital of Capital Medical University were consecutively enrolled.The Hamilton Depression 17-item Scale(HAMD-17)was used to diagnose depression(scores ≥14). The depressive characteristics of PD patients with depression and those without depression were compared, and Logistic regression model was used to analyze the clinical risk factors of depression in PD patients.Results:Among the 1 138 PD patients, 233 cases had depression.The incidence of depression in PD patients was 20.5%, and the treatment rate was only 6.0%(14 cases). The main symptoms of depression in PD patients included sleep disturbance(101 cases, 43.3%), depression(57 cases, 24.5%), work and interest loss(49 cases, 21%). Compared to PD patients without depression, PD patients with depression were more likely women(49.4% vs36.3%), and had lower cognitive scores[(25.56 ± 4.22)scores vs(27.07 ± 3.08)scores], higher rate of freezing of gait(51.0% vs24.0%), higher incidence of disability(87.1% vs28.5%)( P<0.05). A comparison of each part of the unified Parkinson's disease rating scale(UPDRS)between the two groups revealed that the scores of activities of daily living[(16.52 ± 8.71)scores vs(10.15 ± 5.48)scores], the motor examination[(33.78 ± 19.48)scores vs(22.87 ±13.08)scores]and the complications of therapy[2.0(0.0, 6.0)scores vs0.0(0.0, 2.0)scores]were higher in the PD patients with depression group than in the PD patients without depression group( P<0.05). Logistic regression analysis showed that women( OR=1.532, 95% CI: 1.073-2.187, P=0.019), disability( OR=6.357, 95% CI: 4.399-9.186, P<0.001), activities of daily living evaluated by UPDRS( OR=1.093, 95% CI: 1.043-1.146, P<0.001)and motor complications( OR=1.100, 95% CI: 1.014-1.193, P=0.022)were independent risk factors for PD depression. Conclusions:Depression was common in PD patients and usually manifested as sleep disturbance and low motivation, women, motor complications, disability and decreased quality of daily living were independent risk factors for depression.