Objective To detect the expressions of CD70 mRNA and protein and to determine the methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus (SLE).Methods Peripheral blood CD4+ and CD8+ T cells were isolated from 15 patients with active SLE,15 patients with inactive SLE and 15 healthy control subjects.Real-time quantitative reverse transcription-PCR was carried out to quantify the mRNA expression of CD70,flow cytometry to determine the frequency of CD4+CD70+ and CD8+ CD70+ T cells,and bisulfite sequencing to evaluate the methylation status of CD70 gene promoter in CD4+ and CD8+ T cells.Differences in these parameters among these groups were analyzed by one-factor analysis of variance and SNK-q test.Results Compared with the healthy controls,the patients with active SLE and inactive SLE showed a significant increase in CD70 mRNA expression in CD4+ T cells (0.82 ± 0.12 and 0.73 ± 0.11 vs.0.45 ±0.09,F =53.017,P ＜ 0.01) and in the frequency of CD70+CD4+ T cells (80.30％ ± 11.04％ and 66.80％ ± 3.98％ vs.12.48％ ± 3.45％,F =311.517,P ＜ 0.01).Also,the expression of CD70 mRNA in CD4+ T cells and the frequency of CD70+CD4+ T cells were significantly higher in patients with active SLE than in patients with inactive SLE (both P ＜ 0.05).There was a positive correlation between the frequency of peripheral CD70+CD4+ T cells and disease activity in SLE in these patients (r =0.792,P ＜ 0.01).The average methylation index of the region between-600 bp and-300 bp of CD70 gene promoter in CD4+ T cells was 0.32 ± 0.05 and 0.36 ± 0.05 respectively in the patients with active and inactive SLE,significantly lower than that in the healthy controls (0.62 ± 0.05,F =152.64,P ＜ 0.01),and the patients with active SLE showed a significantly lower methylation index than those with inactive SLE (P ＜ 0.05).Conclusions The CD70 gene promoter in CD4+ T cells is significantly hypomethylated in patients with SLE,which may directly lead to the overexpression of CD70.

Objective To study the expression of intestinal dipeptide transporter 1 (PEPT1) protein in gastric cancer patients at different nutritional status,and to explore the possible regulatory mechanism.Methods According to Nutritional Risk Screening 2002 (NRS 2002) score,a total of 60 gastric cancer patients were divided into nutritional risk group (NRS 2002 score ≥ 3,n =18) and non-nutritional risk group (NRS 2002 score ＜ 3,n =42).With specimens of the small intestinal mucosa taken during operation,the expression of intestinal PEPT1 protein was detected using Western blot.The serum concentration of tumor necrosis factor (TNF-o) were measured using enzyme-linked immunosorbent assay.The expression of PEPT1 in Caco-2 cells treated with different concentrations of TNF-α (20,50,100 μg/L) were detected using Western blot at different time points (24,48,72 hours).Results The expression of intestinal PEPT1 (0.63 vs.0.23,P =0.000) and serum TNF-o concentration (0.23 μg/L vs.0.17 μg/L,P =0.001) in the nutritional risk group were significantly higher than those in the non-nutritional risk group.In Caco-2 cells,those treated with different concentrations of TNF-α (20,50,100 μg/L) for 24 hours had significantly higher PEPT1 expression than the blank group did (0.68 vs.0.54,P =0.005 ; 0.72 vs.0.54,P =0.001 ;0.78 vs.0.54,P =0.000).The Caeo-2 cells treated with 50 μg/L TNF-α for 24,48,and 72 hours had significantly higher PEPT1 expression compared with the cells in the blank group (0.57 vs.0.52,P =0.004 ; 0.75 vs.0.52,P =0.000 ; 0.77 vs.0.52,P =0.000).Conclusion The expression of intestinal PEPT1 protein was increased in gastric cancer patients with nutritional risk,which was probably attributed to the regulation of TNF-α.

OBJECTIVE:To virtually screen lignanoid compounds with inhibitory effect of histone deacetylase(HDAC)by vir-tual screening method. METHODS:Using“lignanoid”as keyword,requiring CNKI,VIP,PubMed and other database,lignanoid compounds were collected as ligand to establish ligand base, histone deacetylase receptor HDAC2 (PDB code:4LXZ) and HDAC8 (PDB code:1T69) were selected from PDB database,and then ligands and 3D active site of receptors were docked by SYBYL-X 2.0 software. The affinity of receptors to ligand was reflected by total score. RESULTS:345 lignanoid compounds, 4LXZ and 1T69 primary ligand were used to establish ligand base which included 347 ligands. Ligands No.275,271,110,200, 056,258,181,129,037,270,187 were demonstrated good affinity with receptors HDAC2 and HDAC8. Ligands and receptors residue were docked via hydrogen bond. CONCLUSIONS:Lignanoid compounds have inhibitory effect on HDAC;virtual screen-ing method is effective in natural product activity prediction,which can provide quick access to and theoretical guidance for new pharmacological studies of lignanoid compounds.

Objective We undertook this study to analyze anxiety in tinnitus patients .Methods The partici‐pants were recruited from patients seeking treatment for primary tinnitus and were asked to complete self rating anx‐iety scale(SAS)and Chinese (Mandarin) tinnitus handicap inventory(THI -CM) .Results A total of 224 tinnitus patients were included in the study .The mean score of SAS was 46 ± 2 .55 .The average SAS score among different degrees of anxiety (no ,mild ,moderate ,severe) participants were 65 .63% ,20 .54% ,8 .04% ,5 .80% .The average SAS score for male and female were 46 .35 ± 12 .81 ,46 .28 ± 12 .40 ,respectively .No statistic differences were ob‐served among different genders whenever the degrees of anxiety and different ages (p>0 .01) .The mean SAS scores among young ,middle ,old age groups were 47 .72 ± 12 .80 ,44 .56 ± 12 .83 ,40 .28 ± 7 .24 ,respectively .The pearson correlation between tinnitus duration and the score of SAS was 0 .36(P=0 .594) .The pearson correlation between the scores of THI-CM and SAS was 0 .548(P<0 .01) .Conclusion Most tinnitus participants had mild or didn't have anxiety .Only 13 .84% participants have moderate or severe anxiety .Tinnitus handicap degree increased with the increase of anxiety .No anxiety difference was observed among different sex ,tinnitus duration and age groups . The anxiety of young age group was more severe than that of old groups .

Derivation of human embryonic stem cell and embryonic germ cell lines has widespread and far-reaching significance on human basic research and transplantation therapies. Human pluripotential stem cells provide an exciting new model for studying early human embryogenesis, understanding normal human development and abnormal development, provide a powerful system for discovering human novel genes and testing their function, offer new strategies for discovering of novel growth factors and medicines and promise a renewable source of cells for tissue transplantation, cell replacement and gene therapies. Research history of establishment of human ES and EG cell lines is reviewed. Several methods of establishment of these cell lines involving in the protocol, route, significance and possibility are discussed. Selection of the feeder layer, medium, and supplemental cytokines and their roles in establishing and maintaining human ES and EG cell lines at present are illustrated in detail systematically. Effects and used methods of several kinds of digestive en-zyme in propagations are prepared. Several methods for identifying human ES and EG cells are summarized. At the end, some key problems which are urgent to resolve in these studies at present are put forward and analyzed.
Animals ; Cell Culture Techniques ; methods ; Cell Line ; Embryo, Mammalian ; cytology ; Humans ; Stem Cells

Adolescent ; Child ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Killer Cells, Natural ; pathology ; Leukocyte Common Antigens ; analysis ; Lymphoma, T-Cell ; genetics ; immunology ; Male ; Microtubule-Associated Proteins ; genetics

Objective:To evaluate the role of autophagy in ischemia postconditioning (IPO)-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty-two SPF healthy adult male C57BL/6J mice, aged 9-12 weeks, weighing 25-29 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group (group S), intestinal I/R group (group IIR), group IPO and IPO plus 3-methyladenine (3-MA) group (group IPO+ 3-MA). The model of intestinal I/R was established by occlusion of superior mesenteric artery for 45 min followed by 2-h reperfusion in anesthetized animals.The mice underwent 3 cycles of 30-s reperfusion followed by 30-s ischemia before restoration of reperfusion in group IPO.Blood samples from the femoral artery were collected at 2 h of reperfusion for determination of concentrations of serum diamine oxidase (DAO), D-lactate (D-LA) and intestinal fatty acid binding protein (I-FABP). The animals were then sacrificed and intestinal tissues were removed for microscopic examination of the pathologic changes which were scored according to Chiu and for determination of the expression of autophagy-related proteins Beclin-1 and p62 (by Western blot). The water content of intestinal tissues was calculated. Results:Compared with group S, the Chiu′s score, concentrations of serum DAO, D-LA and I-FABP, water content of intestinal tissues, and LC3Ⅱ/LC3Ⅰ ratio were significantly increased, Beclin-1 expression was up-regulated, and p62 expression was down-regulated in IIR, IPO and IPO+ 3-MA groups ( P<0.05). Compared with group IIR, the Chiu′s score, concentrations of serum DAO, D-LA and I-FABP, water content of intestinal tissues, and LC3Ⅱ/LC3Ⅰ ratio were significantly decreased, Beclin-1 expression was up-regulated, and p62 expression was down-regulated in group IPO ( P<0.05). Compared with group IPO, the Chiu′s score, concentrations of serum DAO, D-LA and I-FABP, and water content of intestinal tissues were significantly increased, LC3Ⅱ/LC3Ⅰratio was decreased, Beclin-1 expression was down-regulated, and p62 expression was up-regulated in group IPO+ 3-MA ( P<0.05). Conclusion:Autophagy is involved in ischemia postconditioning-induced attenuation of intestinal I/R injury in mice.

Objective:To evaluate the relationship between mitochondrial calcium uniporter protein (MCU)-mediated mitochondrial dynamics and intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Twenty-four wild-type adult male C57BL/6J mice, aged 6-8 weeks, weighing 18-20 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (S group), intestinal I/R group (IIR group), sham operation+ MCU inhibitor Ru360 group (S+ Ru360 group) and intestinal I/R + Ru360 group (IIR+ Ru360 group). The mouse model of intestinal I/R injury was prepared by clamping the root of the superior mesenteric artery for 45 min followed by 2 h of reperfusion in anesthetized animals. Small intestinal tissues were obtained at the end of reperfusion for examination of the intestinal mucosal injury which was scored according to Chiu and for determination of the content of malondialdehyde (MDA) (TBA method), activity of superoxide dismutase (SOD) (WST-8 method), content of lactic dehydrogenase (LDH) (colorimetric method), and expression of MCU, dynamin-related protein 1 (Drp1), recombinant human mitochondrial fission 1 protein (Fis1), mitofusin-1 (Mfn1) and Mfn2 (by Western blot). Results:Compared with S and S+ Ru360 groups, the Chiu′s score and contents of MDA and LDH were significantly increased, the activity of SOD was decreased, the expression of MCU, Drp1 and Fis1 was up-regulated, and the expression of Mfn1 and Mfn2 was down-regulated in IIR and IIR+ Ru360 groups ( P<0.05). Compared with IIR group, the Chiu′s score and contents of MDA and LDH were significantly decreased, the activity of SOD was increased, the expression of MCU, Drp1 and Fis1 was down-regulated, and the expression of Mfn1 and Mfn2 was up-regulated in IIR+ Ru360 group ( P<0.05). Conclusions:The mechanism underlying intestinal I/R injury may be related to MCU-induced promotion of mitochondrial fission, reduction of mitochondrial fusion and mediation of imbalance in mitochondrial dynamics in mice.

OBJECTIVETo determine the operation procedure of total hip arthroplasty (THA) for Crowe type-IV developmental dysplasia of hip and its relationship with nerve injury.

METHODSA consecutive series of 39 THAs was performed for Crowe type-IV developmental dysplasia of hip in 35 patients (all female). The mean age of the patients at the time of surgery was 46 years (range 36 - 56 years). Thirty-five hips in 31 patients were followed up. The average follow-up period of the whole series was 4 years (range 1 - 8 years). All procedures were carried out through a lateral-posterior approach. In 33 of 35 hips, the cup was inserted in the "true" acetabulum. All the prostheses used were cementless, except for 5 cemented femoral stems in 5 patients. Each patient was evaluated clinically and by radiographs before the operation and during the follow-up period, according to the Harris hip score (HHS).

RESULTSNone of the cups and stems were revised for aseptic loosening, dislocation or infection during the follow-up period. The mean preoperative HHS was 43 compared with the postoperative HHS of 87. The mean amount of postoperative leg lengthening was 5 cm (range 4 - 6 cm).

CONCLUSIONSThe reconstruction of the hip at the level of the "true" acetabulum through a lateral-posterior approach is a safe and effective procedure of THA for Crowe type-IV developmental dysplasia of hip in adults. Acute leg lengthening of less than 6 cm could not cause nerve injury.

Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Humans ; Middle Aged ; Treatment Outcome

OBJECTIVETo evaluate the safety of stapled transanal rectal resection (STARR) for the treatment of obstructed defecation syndrome(ODS).

METHODSA retrospective study was performed in 112 female patients with ODS eligible for STARR. The short-lerm and long-term postoperative complications were recorded and assessed.

RESULTSShort-term postoperative complications and adverse events were reported in 18 patients (16.1%) including fecal incontinence (4.5%), anastomotic bleeding (2.7%), staple line partial dehiscence (0.9%), anal fissure (2.7%), acute urinary retention (1.8%), thrombosed external hemorrhoid (1.8%), hematoma of the rectovaginal septum (0.9%) and fecal impaction (0.9%). Reoperation was required in 2 patients (1.8%) due to the short-term postoperative complications. The median length of follow-up was 24 months. There were 6 patients with long-term postoperative complications (5.4%) including fecal incontinence (1.8%), defecatory urgency (0.9%), chronic pain due to anastomotic inflammation (1.8%), and chronic pain due to anal rectal diverticulum (0.9%). Three patients (2.7%) were reoperated.

CONCLUSIONSTARR appears to be a safe technique for patients with obstructed defecation.

Defecation ; Digestive System Surgical Procedures ; Female ; Humans ; Postoperative Complications ; Rectal Diseases ; surgery ; Reoperation ; Retrospective Studies ; Surgical Stapling ; adverse effects ; Treatment Outcome