Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Gene Rearrangement, T-Lymphocyte ; Humans ; Lymphoma ; genetics ; Lymphoma, Extranodal NK-T-Cell ; genetics ; Lymphoma, Large-Cell, Anaplastic ; genetics ; Lymphoma, T-Cell, Peripheral ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Young Adult

Objective To investigate the effects of the folic acid on human T lymphoid leukemia cell line CEM cells. Methods 1. MTT method was used to detect the proliferation of CEM cells co- cultured with folic acid of different concentrations and time;2. E-xamine the changes of morphology by light microscopy with Giemsa stain;3. Detect the percentage of apoptosis and cell cycle distribution as well as the expression of the apoptosis protein(Bcl- 2,C- myc) by flow cytometry;4. Detect DNA fragments by Agaiose elec-trophoresis;5. Detect the influence of folic acid to the anticancer effects of methotrexatc(MTX) by MTT methods. Results 1 Folic acid could inhibit the proliferation of CEM cells, and the optimal inhibitive concentrations range from 0. 4 ? 10-4 ?g/L to 3. 0 ? 10 -4 ?g/L,the inhibition rate was about 30% - 40% ; 2. Co - cultured with folic acid at 24,48, 72 hours, examined by light microscopy with Giemsa stain, apoptosis cells were found in all study groups but the higher apoptosis rate was found co - cultured with folic acid at concentration of (0.4 - 3.0) ? 10-4?g/L;3.The highest apoptosis rate was 6. 19% found at the concentration of 3 ? 10-4 ?g/L, but the cell cycle distribution had no statistical difference with control group, the expression of apoptosis related protein Bcl - 2 and C-myc was decreased;4 DNA was extracted from CEM cells co - cultured with 0.4? 10 -4 ?g/L and 3 ? 10-4 ?g/L folic acid for 48 hours. UNA ladders were visible by agarose electrophoresis of DNA fragments; 5. Folic acid did not affect the antitumor effect of MTX at the concentration from 0 2? 10-4 ?g/L to 12.0?10-4 ?g/L. Conclusion Folic acid may suppress proliferation and induce apoptosis of CEM cell

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

Forensic Medicine ; Humans ; Kidney Transplantation ; Polycystic Kidney Diseases

Objective To investigate the correlation between family cohesion and adaptability and quality of life in patients with enterostomy. Methods Using Chinese version of Family Adaptability and Cohesion Scale (FACESⅡ-CV) and Chinese version of Stoma-Quality of Life (STOMA-QOL-C) to investigate the status of family cohesion and adaptability, family type and their impact on quality of life of 110 patients with enterostomy. Results Scores of family cohesion and adaptability averaged 59.15 ± 11.94, 47.32 ± 9.40,were significantly lower than 63.90 ± 8.00 and 50.90 ± 6.20 in the norm,and the difference was statistically significant (t=-4.171,-3.990, P<0.01).The family cohesion was positively correlated with the score of quality of life, social interaction and psychological burden(r=0.274, 0.284, 0.263, P<0.05), and the family adaptability was positively correlated with the score of quality of life,social interaction and psychological burden(r=0.316, 0.338, 0.228, P<0.05 or 0.01). The balance type family was 30 cases;scores of quality of life averaged 45.10±7.26, the intermediate type family was 50 cases;scores of quality of life averaged 43.48±9.98, the extreme type family was 28 cases;scores of quality of life averaged 43.37 ± 16.68, and difference between the three was no statistically significant(F=0.442, P=0.665). Conclusions In the nursing process of patients with enterostomy, health care workers should pay attention to improve family cohesion and adaptability, as to achieve the purpose of improving the quality of life of the patients.

DNA, Neoplasm ; Humans ; Image Cytometry ; Pleural Effusion, Malignant ; diagnosis

The present study aimed to investigate the role of spinal p38 mitogen-activated protein kinase (p38 MAPK) activation in chronic constriction injury (CCI) of the sciatic nerve induced neuropathic pain. CCI model was produced by loosely ligating the left sciatic nerve proximal to the sciatica's trifurcation with 4-0 silk thread in male Sprague-Dawley rat. SB203580, a specific inhibitor of the p38 MAPK, was intrathecally administered on day 5 post-CCI. Thermal and mechanical nociceptive thresholds were assessed with the paw withdrawal lantency (PWL) to radiant heat and the paw withdrawal threshold (PWT) to von Frey filaments respectively. The protein levels of the phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated cAMP response element binding protein (pCREB) were assessed by Western blot analysis. The results showed that CCI significantly increased the expressions of cytosolic and nuclear p-p38 MAPK in the spinal cord. Intrathecal administration of SB203580 dose-dependently reversed the established mechanical allodynia and thermal hyperalgesia induced by CCI. Correlated with behavior results, SB203580 dose-dependently inhibited the CCI-induced increase of the expressions of cytosolic and nuclear p-p38 MAPK and nuclear pCREB in the spinal cord. Taken together, these findings suggest that the activation of p38 MAPK pathway contributes to the development of neuropathic pain induced by CCI, and that the function of p-p38 MAPK may partly be accomplished via the CREB-dependent gene expression.
Animals ; Female ; Ligation ; Male ; Neuralgia ; enzymology ; etiology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; Spinal Cord ; enzymology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the difference of postoperative drainage and systemic trauma response between endoscopic and traditional near total thyroidectomy to provide the basis for selecting appropriate operative methods.

METHODSIn this prospective clinical controlled study, 80 patientsscheduled for near total bilateral thyroidectomy for the first time were divided equally into endoscopic surgery group (group A) and open surgery group (group B). The total drainage volume after operation, postoperative extubation time, and postoperative daily drainage volume were recorded after the operation. The contents of triglyceride (TG) and total protein (TP) were determined in the postoperative drainage fluid onthe first day. The levels of interleukin 6 (IL6), high sensitive C reactive protein (HSCRP), alpha 1 acid glycoprotein (AAG), ceruloplasmin (CER) and haptoglobin (HPT) in venous blood were tested before the operation and on the first day after surgery.

RESULTSCompared with those in group B, the postoperative drainage volumein group Aincreased significantly (P=0.000) and the postoperative extubation time was significantly prolonged (P=0.000); the mean postoperative daily drainage volume was significantly larger ingroup A than in group B (P=0.000) and tended to decrease with time in both groups. There was no significant difference in the content of triglycerideortotal protein in the drainage fluid between the two groups on the first day after operation (P=0.429 and 0.324, respectively). In both groups, the contents of AAG, ceruloplasmin and haptoglobin on the first postoperative day were all similar with those measurement before operation (P>0.05), but significant variations occurred in the levels of IL6 and HSCRP on the first postoperative day (P=0.000). The serum levels of IL?6 or HS?CRP did not differ significantly between the two groups on the first day after operation (P=0.054 and 0.066, respectively).

CONCLUSIONCompared with open surgery, endoscopic near total bilateral thyroidectomyis associated with an increased the volume of postoperative drainage and a prolonged time of extubationbut not an increased systemic trauma response. Therefore, endoscopic surgery can serve as one of the routine options for patients who are concerned with neckscars resulting from open surgeries.

OBJECTIVETo investigate the gene defect in a hereditary coagulation factor V (FV) deficiency family.

METHODSThe plasma FV actigen was measured by one-stage clotting assay. The FV antigen was assayed by Biotin-Avidin enzyme linked immunosorbent assay (BA-ELISA). The full length of exon 1 to exon 25 and the 5' untranslated sequence of FV genomic DNA were analyzed by polymerase chain reaction (PCR) and direct sequencing of the amplified fragments, meanwhile the defect was identified by T/A cloning sequencing.

RESULTSThe plasma coagulant activity and amount of FV of the proband were marked deficient (1% and 1.54%, respectively). DNA sequence analysis for the proband revealed a causative mutation in a heterozygous status. It was one base pair deletion in exon 4 at nucleotide 675 inherited from her mother.

CONCLUSIONSA novel mutation in the FV gene was identified in the proband with congenital FV deficiency. The mutation was 675delA in exon 4 resulting in a frameshift and a premature termination codon.

Adolescent ; Blood Coagulation ; Factor V ; analysis ; genetics ; Factor V Deficiency ; blood ; genetics ; Female ; Humans ; Mutation

OBJECTIVETo investigate effect of oxidized low-density lipoprotein (ox-LDL) on memory CD8T cell subpopulation differentiation in mice with autoimmune diabetes.

METHODSCultured splenic CD8T cells from pre-diabetic NOD mice isolated with magnetic beads were treated with 30 µg/mL ox-LDL and 10 U/mL interleukin-2 (IL-2) for 24 h and the control cells were treated with IL-2 only. Flow cytometry was used to determine the percentage of splenic CD8IFN-γT cells, expressions of CD8, CD44 and CD62L on the T cells, and the activation of T cell factor-1 (TCF-1) and STAT-3. The CD127memory T cells were purified and transplanted into the pre-diabetic NOD mice via the tail vein, and the blood glucose was recorded weekly and survival time of the mice was monitored.

RESULTSTreatment with ox-LDL significantly reduced islet β cell-specific cytotoxic CD8T cells as compared with the control group [(0.7∓0.03)% vs (2.7∓0.14)%, P<0.01]. The percentage of effector memory CD8T cells (Tem) in the total memory CD8T cells was reduced [(10.3∓0.71)% vs (30.3∓1.36)%, P<0.01] and that of stem cell-like memory T cells was significantly increased [(72.3∓3.8)% vs (55.1∓2.61)%, P<0.05] following ox-LDL treatment, which also resulted in significantly decreased activation of TCF-1 [(14.5∓0.82)% vs (34.2∓1.23)%, P<0.01] and pSTAT-3 [(3.3∓0.12)% vs (22.1∓1.1)%, P<0.01]. Transplantation of ox-LDL-treated memory T cells in pre-diabetic NOD mice obviously inhibited the increase of blood glucose and prolonged the survival time of the mice (P<0.05).

CONCLUSIONOx-LDL decreases the activation of transcriptional factors TCF-1 and phosphorylation of STAT-3, inhibits the formation of effector memory CD8T cells with long-term cytotoxicity, but promote the generation of stem cell-like memory CD8T cells, which result in suppression of islet β cell-specific effector cytotoxic CD8T cell differentiation to lessen autoimmune injury to the islet β cells.