Objective To investgate the expression of Tfh and Th9 in the peripheral blood of rheumatoid arthritis ( RA),and to analyze the relationship between those cells and disease activity,organs of involvement,ect.,and to underly the possible immunological mechananism of RA.Methods Peripheral blood Tfh cells and Th9 cells,define as the CD4+ CXCR5 + ICOS+ and CD3 + CD8+ IL-9+ population,were examined by flow cytometry in 36 patients with RA and 22 case of age-and gender-matched health controls.According to 28-joint Disease Activity Score (DAS28),patients were divied into two groups:high disease activity(n=22),moderate disease activity(n=14).And then to analyzed the correlation among the Tfh,Th9 and clinical data of RA,such as ESR,CRP,RF,the number of tender joints,the number of swollen joints and bone destruction,etc..While to analyzed the correlation between Th9 and Tfh.Results The expression of Tfh (Z=-6.082,P=0.000) and Th9 (0.989±0.498 vs 0.213±0.084,t=13.063,P =0.000 )in PBMCs of RA was significantly higher than normal controls.Meanwhile,the expression of Tfh ( 3.880 ±1.255 vs 2.678±1.022,t=2.990,P=0.005 ) and Th9( 1.181 ±0.523 vs 0.686±0.254,t =4.043,P=0.000) in high activity of group were higher than those moderate.The percentage of Tfh and Th9 in the two groups were all signifieantly higher than controls(P＜0.01 ),There was a positive correlation with Tfh and DAS28 ( r =0.571,P=0.000),ESR ( r =0.375,P =0.029),CRP( r =0.357,P =0.032 ),the number of tender joints ( r=0.598,P =0.000 ),RF ( r =0.421,P =0.023 ) and anti-CCP ( r =0.421,P =0.023 ).There were no relationship with Tfh and course,morning stiffness,the number of swollen joints,bone erosion and the abnormal of EKG.There was a positive correlation with Th9 and DAS28( r=0.461,P=0.005 ),ESR(r=0.347,P=0.042),CRP(r=0.384,P=0.210),the number of tender joints(r=0.341,P=0.042),the number of swollen joints (r =0.347,P=0.038 ) and RF( r =0.379,P =0.025 ) respectively.There were no relationship with Th9 and course,morning stiffness,CCP,the abnormal of EKG,bone erosion.There was a positive correlation with Tfh and Th9 (r=0.727,P =0.000),Conclusion The expression of Tfh and Th9 were increase significantly,which those cells were close relationship with the disease activity and some data,These results indicate that the abnormality of Tfh and Th9 may play an important role in the pathogenesis of RA.

Objective To detect the expression of T helper cells 9 (Th9) in the peripheral blood and the level of interleukin (IL)-9 in the plasma of rheumatoid arthritis (RA) patients,and to analyze the relationship between Th9 and clinical characteristics,and their possible role in the immunological pathogenesis of RA were explored.Methods Thirty-six patients with RA and 22 healthy controls were included.RA patients were divided into high disease activity group (n=22) and moderate disease activity group (n=14).Th9 cells were obtained from RA patients and healthy controls and detected by flow cytometery.The levels of IL-9 in RA patients and healthy controls were detected by ELISA tests.T-test and Spearman's correlation were used for statistical analysis.Results The expression of Th9 and the level of IL-9 of RA patients [(0.99±0.50)％,( 1.06±0.42 ) pg/ml,respectively] were significantly higher than normal controls[ (0.21±0.08)％,(0.69±0.13)pg/ml,respectively,all P＜0.01 ].Meanwhile,the expression of Th9 and the level of IL-9 in high disease activity group [ ( 1.18±0.52 )％,( 1.37±0.47 ) pg/ml ] were higher than those of the moderate disease activity group [ (0.69±0.25)％,(0.94±0.30) pg/ml ].There were positive correlations between Th9 and disease activity score(DAS) 28,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),the number of tender joints,the number of swollen joints and rheumatoid factor (RF),the level of IL-9 respectively.There were positive correlations between IL-9 and DAS28,ESR,the number of tender joints,and the number of swollen joints.Conclusion High expression of Th9 and level of IL-9 maybe involved in the pat-hogenesis of RA,and is associated with disease activity.These results indicate that the abnormality of Th9 may play an important role in the pathogenesis of RA.

Objective To test the level of cell factor interleukin (IL)-21,CXCL13 in the plasma of patients with rheumatoid arthritis (RA),and to analyze the relationship between Follicular helper T cells(Tfh)and clinic features and discuss the possible immunological pathogenesis of RA.Methods The Tfh cells were obtained from patients and healthy controls (NC) and detected by Flow cytometery.While the levels of IL-21,CXCL13 in patients and NC were measured by ELISA tests.Those analysis were performed by student's t-test,one-way ANOVA,SNK-q test,Chi-square test,Spearman's correlation and multiple linear regression.Results The expression of CD4+CXCR5+ICOS+ cells (Tfh) in PBMCs of RA was significantly higher than normal controls (3.0±1.2 vs 1.1±0.4,P＜0.01).Meanwhile,the three RA groups of patients were divided to low,moderate and high disease activity groups,and the results showed that the expression of Tfh were increased accordingly (1.8±0.7,2.5±0.6,4.0±1.2).The expression of Tfh in the three groups were all significantly higher than that of controls (P＜0.01).There was a positive correlation between Tfh and DAS28,ESR,CRP,TJC,and bone erosion,RF and anti-CCP respectively.The expression of Tfh in those patients who had bone destruction was higher than those with no or mild bone destructions (2.7±1.1vs 3.4±1.3).The expression of Tfh in patients with un-treated RA patients,when compared to those RA patients who were treated appropriately and those who were not treated appropriately,was decreased significantly.The expression of Tfh in appropriately treated RA patients was lower than that without appropriately treatment.The level of IL-21,CXCL13 was decreased in patients with RA in the order of high,moderate,low disease activity and NC.Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,and are closely related to disease activity and bone ersions.The expression of Tfh is decreased after relevant treatment.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of RA.

To screen antitumor active compounds, drug-like or leading compounds from Chinese traditional and herbal drugs.

Objective:To construct a cognitive training program suitable for elderly patients with mild cognitive impairment based on horticultural therapy, so as to effectively slow down the cognitive decline of patients with mild cognitive impairment.Methods:Through searching the Chinese and English database literature of cognitive intervention from July 2000 to July 2020 and field visits to nursing homes, the draft intervention plan was formed. Two rounds of focus group interview were held to consult experts in cognitive impairment and geriatric care, etc., and to revise the intervention plan.Results:In the two rounds of focus group interview, the expert positive coefficient was 100%, the expert judgment basis was 0.84, the expert familiarity degree was 0.84, and the expert authority coefficient was 0.84. In the end, a 10-week cognitive intervention program targeting six cognitive domains -- "visuospatial/executive ability", "memory ability", "language ability", "attention ability", "abstract ability" and "naming ability" was formed, and the implementation steps of the program were improved.Conclusions:The construction process of cognitive training program for patients with mild cognitive impairment based on horticultural therapy theory is rigorous, scientific and feasible, and can be used to guide the cognitive training of patients with mild cognitive impairment.

Objective To introduce an optimized practical method of making and detecting pipettes for microinjection.Methods Transfer pipette was made from hard glass capillary. We softened the hard glass capillary by rotating it in a spirit-lamp flame,then moved out from the flame and quickly pulled it into two transfer pipettes.After broken by a grinding wheel,the tip of the pipette was fire-polished by quickly touching the flame to make a fine opening.A hard glass capillary (1.0 mm,ouside diametre)was pulled into two holding pipettes by pipette Puller.The pipette shoulder was broken at desired position with a grinding wheel,then the fine pipette tip opening was heated by a microforge and shrinked into a diameter -15 μm.Injection pipette could be made directly from a capillary with filament by Puller.The solution loaded injection pipette and holding pipette were assembled into the micromanipulator and could be checked before use.We transfered both pipettes into the zygotes media drop,touched the holding pipette with the tip of the injection pipette to make a "suitable"opening.Then we switched injection pipette to the mineral oil and applied injection pressure through the injector to check whether the solution could come out of the tip smoothly and at a proper speed.It could be further verified by pronucleus microinjection of zygotes.Results The results showed that the method introduced in this paper could produce suitable pipettes for zygote microinjection.In particular,the method of detecting the opening of the injection pipette was helpful for achieving high efficiency of zygote microinjection.Conclusion The method introduced here to make and detect pipettes for microinjection is very helpful for establishing a standard microinjection manipulation procedure and improving the efficiency of zygote microinjection.

Neural recording electrodes enable the acquisition and collection of electrical signals from neu-rons,and these recorded neural electrical signals are an important means of understanding neuronal activity.As a major component of the brain-machine interface,neu-ral recording electrodes serve as a bridge between the nervous system and external devices.The extracted information can be used to understand the state of the brain and acts as a feedback signal to regulate external devices,thus providing important information for the clini-cal treatment of neurological diseases.Moreover,the electrodes can be used as a vehicle for drug injection to directly treat diseases.Since the time that Strumwas-ser used microwires to achieve long-term recordings of neural activity in hibernating squirrels,implantable elec-trode technology has gradually improved over three gen-erations of development,and progress has been made in improving the biocompatibility,mechanical performance(size,shape,density,etc.),and signal-to-noise ratio.Implantable neural recording electrodes can acquire sig-nals from cortical and deep neural clusters,with the advantages of high signal-to-noise ratio,information con-tent,and spatial/temporal resolution.However,there is still a need to improve the structure and performance of these electrodes;for example,their high invasiveness and lack of biocompatibility pose technical difficulties in the process of translation to the clinic.This paper reviews the basic requirements for electrodes,main recording methods and signal types,common types of implant-able neural recording electrodes,and their challenges and future development directions.With the continuous development of electrode materials,equipment,systems,and neurotechnology,it should be possible to apply neu-ral recording electrodes in clinical practice,to promote safe and efficient treatment of human diseases.

Objective To explore the preparation of a rat model of pneumonia model induced by Pseudomonas aeruginosa( PA) using different methods,and to lay the foundation for further studies. Methods 48 SD rats were randomly divided into 4 groups:the control group (A), the intratracheal injection group (B), the trachea cannulation group (C) and the intranasal inoculation group ( D) . After intervention with different treatment modalities, the body weight,tempera-ture,white blood cell count and lung pathological changes in the rats of all groups were detected at 5, 10, 15 days. Results 1. The behavior, body weight, temperature, leukocytes and pathological inflammatory changes of the lung in rats of the model groups were significantly different from that of control group. 2. Pseudomonas aeruginosa was detected in rats of all the model groups, but the control group was negative. Conclusions Rat model of Pseudomonas aeruginosa infected pneu?monia can be successfully established by intranasal inoculation. This method can avoid the inflammatory interference from operation, and is simple and suitable for popularization.

The aim of this study was to identify molecular markers associated with oncogenic differentiation in hepatocellular carcinoma (HCC). Using an unsupervised clustering method with a cDNA microarray, HCC (T) gene expression profiles and corresponding non-tumor tissues (NT) from 40 patients were analyzed. Of total 217 genes, 72 were expressed preferentially in HCC tissues. Among 186 differentially regulated genes, there were molecular chaperone and tumor suppressor gene clusters in the Edmondson grades I and II (GI/II) subclass compared with the liver cirrhosis (LC) subclass. The Edmondson grades III and IV (GIII/IV) subclass with a poor survival (P = 0.0133) contained 122 differentially regulated genes with a cluster containing various metastasis- and invasion-related genes compared with the GI/II subclass. Immunohistochemical analysis revealed that ANXA2, one of the 72 genes preferentially expressed in HCC, was over-expressed in the sinusoidal endothelium and in malignant hepatocytes in HCC. The genes identified in the HCC subclasses will be useful molecular markers for the genesis and progression of HCC. In addition, ANXA2 might be a novel marker for tumor angiogenesis in HCC.
Annexin A2/genetics ; Carcinoma, Hepatocellular/*genetics/pathology ; Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; Liver Neoplasms/*genetics/pathology ; Molecular Chaperones/genetics ; Multigene Family ; Oligonucleotide Array Sequence Analysis ; Oncogenes ; Tumor Markers, Biological/*genetics

OBJECTIVETo detect the presence of endothelial injury in patients with severe acute respiratory syndrome (SARS) via enhanced levels of tissue-type plasminogen activator (t-PA) and soluble thrombomodulin (sTM).

METHODSCase patients were from Xuanwu Hospital (Capital University of Medical Sciences, Beijing, China), and all of them met clinical criteria for SARS. Healthy controls were some of the hospital employees. Endothelial injury bio-markers tPA and sTM were detected by commercial ELISA-methods.

RESULTSClassic plasma markers of endothelial injury, tPA and sTM significantly elevated in SARS patients in comparison to controls [t-PA: 1.48 +/- 0.16 nmol/L versus 0.25 +/- 0.03 nmol/L (P<0.0001), and sTM: 0.26 +/- 0.06 nmol/L versus 0.14 +/- 0.02 nmol/L (P<0.05)]. The only patient who died had extremely high levels of these endothelial injury markers (t-PA: 2.77 nmol/L and sTM: 1.01 nmol/L). The likelihood ratio analysis indicated the excellent discriminating power for SARS at the optimal cut-point of 0.49 nmol/L for tPA and 0.20 nmol/L for sTM, respectively. Significant numerical correlations were found among these endothelial injury markers in SARS patients. The numerical coefficient of correlation Pearson r between t-PA and sTM was 0.5867 (P<0.05).

CONCLUSIONIncreased plasma concentrations of tPA and sTM in patients with SARS suggest the possibility of endothelial injury. SARS patients might need anticoagulant therapy or fibrinolytic therapy in order to reverse intraalveolar coagulation, microthrombi formation, alveolar and interstitial fibrin deposition. It may not only provide a useful treatment and prognostic index but also allow a further understanding of the pathological condition of the disease.

Adult ; Biomarkers ; blood ; Case-Control Studies ; China ; Female ; Humans ; Male ; Prognosis ; Severe Acute Respiratory Syndrome ; blood ; Thrombomodulin ; blood ; Tissue Plasminogen Activator ; blood