Objective To study B lymphocyte subsets(na(?)ve B cells,memory B cells and plas- mablasts)of peripheral blood in patients with rheumatoid arthritis(RA)and its relationship with autoantibod- ies and clinical manifestation.Methods Blood samples and clinical data of 60 patients with RA were enrolled into this study.They were divided into three groups:active,inactive and refractory RA based on clinical mani- festations and 24 healthy controls were included.CD19 and CD27 of B cells in peripheral blood of RA patients and healthy controls were detected using flow cytometry at single-cell level.Frequence of na(?)ve B cells (CD19~+CD27~-),memory B cells(CD19~+CD27~(dim)),plasmablasts(CD19~+CD27~(high))and average fluorescence in- tensity of CD19 were analyzed,and their relationship with clinical manifestations and rheumatoid factor(RF), anti-typeⅡcollagen(anti-CⅡ),anti-cyclic citrullianted peptide(CCP)antibodies were investigatied.Results Frequence of na(?)ve B cells and plasmablasts in peripheral blood of patients with RA was increased compared with normal control.In contrast,memory B cells in patients with RA were decreased.The na(?)ve B cells subset in inactive and refractory RA was higher than that of healthy controls(P＜0.05),and the memory B cells subset in those groups was lower than that of healthy controls(P＜0.05).The plasmablasts in active and refractory groups of RA were higher than those of healthy controls(P＜0.05).The average fluorescence intensity of CD19 in peripheral blood in patients with RA was positively correlated with ESR,C-reactive protein(CRP),healthy assessment questionaire(HAQ),and plasmablasts was positively correlated with arthrocele index.Na(?)ve B cells,memory B cells and plasmablasts subsets had no relation with RF,anti-CⅡand anti-CCP antibodies. Conclusion B cell subsets in peripheral blood of patients with RA are significantly abnormal,characterized by expanded naive B cells and plasmablasts but diminished memory B cells.Plasmablasts are increasesd in active and refractory groups of RA,and have positive correlation with swollen joint index.B cells may play an important rote in the pathogenesis of RA.

Objective:To investigate the m echanism of D- amino acid oxidase/D- Alanine system in killing K5 6 2 e cells in vitro.Methods:The killing effects of D- Ala on K5 6 2 e cells stably expressing DAAO and GFP were observed.H2 O2 production by DAAO+ cells were m easured by the phenol red oxidation assay.L owry method was used to determ ine the protein quantities of cells and fluorescent intensities of GFP+ cells were assayed by flow cytom eter.Results:KDf Gd cells were killed completely after treated with 2 5 mm ol/L D- Ala for 2 4 h.The effect of D - Ala at 2 0 m mol/L on KDf Gd cells increased apparently within 4 8h,but the same effect was not observed if D - Ala was below 15 m mol/L .The cytotoxicity of D- Ala in KDf Gd cells was more sensitive than in parental K 5 6 2 e cells.The H2 O2 levels in the medium were consistent with the killing effects of D- Ala.Conclusion:The killing effects of DAAO/D- Ala system is m ediated by H2 O2 . [

Objective To explore the clinical significance of preoperative ultrasound-assisted drawings in location of thyroid micronodule resection .Methods A total of 88 patients ( 137 nodules ) who underwent thyroid micro-nodule resection were enrolled in the prospective randomized controlled study .All patients were randomly divided into two groups:46 patients (68 micronodules) in experimental group and 42 patients (69 micronodules) in control group.Inclusion criteria:the maximum diameter of nodule≤1.0 cm. Preoperative thyroid ultrasound was conducted .Patients in experimental group also underwent ultrasound-assisted location of thyroid micronodule .Results The diagnostic accuracy rate of US was 81.82%(69/88) in all patients .With the help of ultrasound-assisted location , all nodules in experimental group were found quickly and accurately.The resection rate of experimental group was 100%(46/46).Whereas 4 nodules (in 4 patients ) were missed in control group with a resection rate of 90.5% (38/42).The postoperative US examinations after 3 months showed that all nodules were completely removed in experimental group while 4 nodules retained in control group .Conclusions Preoperative ultrasound-assisted drawings in location of thyroid micronodule plays an important role in thyroid nodule resection .It is great value of clinical application .

BACKGROUNDBipolar electro-coagulation has a reported efficacy in treating epilepsy involving functional cortex by pure electro-coagulation or combination with resection. However, the mechanisms of bipolar electro-coagulation are not completely known. We studied the acute cortical blood flow and histological changes after bipolar electro-coagulation in 24 patients with intractable temporal lobe epilepsy.

METHODSTwenty-four patients were consecutively enrolled, and divided into three groups according to the date of admission. The regional cortical blood flow (rCBF), electrocorticography, the depth of cortex damage, and acute histological changes (H and E staining, neuronal staining and neurofilament (NF) staining) were analyzed before and after the operation. The t-test analysis was used to compare the rCBF before and after the operation.

RESULTSThe rCBF after coagulation was significantly reduced (P < 0.05). The spikes were significantly reduced after electro-coagulation. For the temporal cortex, the depth of cortical damage with output power of 2-9 W after electro-coagulation was 0.34 ± 0.03, 0.48 ± 0.06, 0.69 ± 0.06, 0.84 ± 0.09, 0.98 ± 0.08, 1.10 ± 0.11, 1.11 ± 0.09, and 1.22 ± 0.11 mm, respectively. Coagulation with output power of 4-5 W completely damaged the neurons and NF protein in the molecular layer, external granular layer, and external pyramidal layer.

CONCLUSIONSThe electro-coagulation not only destroyed the neurons and NF protein, but also reduced the rCBF. We concluded that the injuries caused by electro-coagulation would prevent horizontal synchronization and spread of epileptic discharges, and partially destroy the epileptic focus.

Adult ; Electrocoagulation ; methods ; Epilepsy ; surgery ; Epilepsy, Temporal Lobe ; surgery ; Female ; Humans ; Male ; Temporal Lobe ; surgery ; Young Adult

The internal ribosome entry site (IRES) sequence was derived from encephalomyocarditis virus. It allows to translate two open reading frames at one mRNA, so two genes conjoined by IRES have the same expression rate. K(DfGC) and K(DfGd) cell lines, stably expressing D-amino acid oxidase (DAAO) gene and green fluorescence protein (GFP) genes, were obtained by transfection of K562e cells with retroviral vector pLDfG containing IRES sequence, DAAO cDNA and GFP gene. Fluorescence positive rate and fluorescence intensity of the two cell lines were measured with flow cytometry. H(2)O(2) production by K(DfGC) and K(DfGd) cells treated with D-alanine was measured by the phenol red oxidation assay. The fluorescence positive rate and fluorescence intensity in K(DfGC) and K(DfGd) cell were 94.64% and 96.31% and 202 units and 174 units per 2 x 10(4) cells, respectively. There was exponential correlation between fluorescence intensity and H(2)O(2) level. The above-mentioned results demonstrate that DAAO gene and GFP gene were simultaneously expressed in K562e cell line by the regulation of IRES sequence, and DAAO level was correlated with fluorescence intensity of GFP.
Binding Sites ; genetics ; D-Amino-Acid Oxidase ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; Humans ; Hydrogen Peroxide ; metabolism ; K562 Cells ; Luminescent Proteins ; genetics ; metabolism ; Retroviridae ; genetics ; Ribosomes ; metabolism ; Transfection

BACKGROUND: Adipose-derived mesenchymal stem cells (ADMSCs) have been reported to improve wound healing. However, type I collagen secreted by ADMSCs will contribute to scar formation. Therefore, inhibiting type I collagen secretion from ADMSCs will strengthen its clinical application. OBJECTIVE: To investigate the effect of 1,25(OH)2D3on secretion of type I collagen by ADMSCs and its mechanism. METHODS: Human ADMSCs were isolated by collagenase digestion, and identified by flow cytometry. ADMSCs at passage 4 were cultured in DMEM/F12 medium containing different concentrations of 1,25(OH)2D3(10-7, 10-8, 10-9, 10-10and 0 mol/L) respectively for 4 days. Then, the concentration of type I collagen in cell supernatant was measured by ELISA. Real-time PCR and western blot were used to detect the expression of Smad3 at mRNA and protein levels and phosphorylated protein Smad3 level in ADMSCs cultured with and without 1,25(OH)2D3. To analyze the contribution of Smad3 to the effect of 1,25(OH)2D3, Smad3 inhibitor was added to culture medium 30 minutes before adding 1,25(OH)2D3, and type I collagen in cell supernatant was detected by ELISA at 4 days after addition of SMAD3 inhibitor. RESULTS AND CONCLUSION: 1,25(OH)2D3inhibited the secretion of type I collagen by ADMSCs in a dose-dependent manner. The results of real-time PCR and western blot showed that the expression of Smad3 was upregulated by 1,25(OH)2D3, and the results of western blot showed that the phosphorylated Smad3 protein level in ADMSCs was significantly increased by 1,25(OH)2D3. Moreover, the inhibition of type I collagen secretion by 1,25(OH)2D3could be blocked by Smad3 inhibitor. These results indicate that 1,25(OH)2D3can inhibit the secretion of type I collagen from ADMSCs by up-regulating the expression of Smad3.

OBJECTIVETo study the effects of potassium salts on direct induction of microtubers from different explants in Pinellia ternata.

METHODLeaves, petioles and tubers were cut and cultured on the medium with different kinds of potassium salts and plant growth regulators.

RESULT AND CONCLUSIONLow concentration of potassium salts, which were lower than 4.02 mmol x L(-1), could promote the microtuber formation in vitro. While high concentration of potassium salts, which were more than 12.06 mmol x L(-1), inhibited the formation of microtubers.

Culture Media ; Dose-Response Relationship, Drug ; Nitrites ; administration & dosage ; pharmacology ; Phosphates ; administration & dosage ; pharmacology ; Pinellia ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; growth & development ; Plant Tubers ; growth & development ; Plants, Medicinal ; growth & development ; Potassium Chloride ; administration & dosage ; pharmacology ; Potassium Compounds ; administration & dosage ; pharmacology ; Tissue Culture Techniques

OBJECTIVETo investigate the association between rs142167, rs7216231 single nucleotide polymorphism (SNP) in Wnt3 and nonsyndromic cleft lip and palate (NSCL/P) in Hui and Han population of Ningxia Autonomous Region.

METHODSThe study consisted of 371 NSCL/P patients from Ningxia Hui and Han population (Han population 166, Hui population 205), their parents (196 fathers, 224 mothers, 150 trios) and 258 normal controls (Han population 190, Hui population 68). Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) was used to identify rs142167, rs7216231 genotypes of the samples. The data was analyzed by case-control analysis, transmission disequilibrium test (TDT) and family based associated test (FBAT).

RESULTSCase-control study showed that no differences in cleft lip, cleft palate, cleft lip and palate, and the total case group compared with the control group at rs142167 and rs7216231 (P > 0.05) in Hui and Han population and in stratified comparison. TDT test showed that rs142167 and rs7216231's allele had not over-transmitted (P > 0.05) in NSCL/P. FBAT test showed that G-G specific haplotypes showed statistically significant (P < 0.05).

CONCLUSIONWnt3 gene polymorphism is not relevant with NSCL/ P in Ningxia Hui and Han population.

Brain ; abnormalities ; Case-Control Studies ; Cleft Lip ; Cleft Palate ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide

Objective To evaluate the predicting value of Wells,Kahn,St.André and Constans scores for the diagnosis of deep venous thrombosis in Chinese patients.Methods A total of 274 patients suspected with deep venous thrombosis was prospectively blinded evaluated with the four clinical-score systems.Sensitivity,specificity,positive predictive value,negative predictive value and receiver operation curves were calculated for four clinical scores according sonography results.Results Sonography evidenced deep venous thrombosis in 88 out of 274 patients.The sensitivity,specificity,positive predictive value and negative predictive value was 77.3％,65.6％,51.5％ and 85.9％,respectively,for Wells score; 58％,55.9％,38.3％ and 73.8％,respectively,for Kahn score; 64.8％,55.4％,40.7％ and 76.9％,respectively,for St.André score and 86.4％,37.6％,39.6％ and 85.4％,respectively,for Constans score.ROC was 0.761 for Wells score,which was similar as that of Constans score (0.759),then followed by St.André score (0.627) and Kahn score (0.591 ).Conclusion Our results showed that Wells score and Constans score are superior to Kahn score or St.André score for diagnosing patients with suspected deep venous thrombosis in terms of sensitivity,negative prediction value and ROC values.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Hepatitis B ; epidemiology ; Humans ; Infant ; Middle Aged ; Seroepidemiologic Studies ; Young Adult