Objective To study the immunosuppressive mechanism of alkaloid sinomenine (SIN) by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells (NF-AT) and interferon-gamma (IFN-γ) in CD4+ T lymphocytes of human periphery blood. Methods CD4+ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. (1) Negative control group: no medicine was added to cell culture medium; (2) Positive control group: CsA solution (final concentration: 50ng/ml) was added to cell culture media; (3) Low-concentration SIN group (L-SIN): low-concentration SIN solution (final concentration: 10 μmol/L) was added to cell culture media; (4) Middle-concentration SIN group (M-SIN): middle-concentration SIN solution (final concentration: 200 μmol/L) was added to cell culture media; (5) High-concentration SIN group (H-SIN): high-concentration SIN solution (final concentration: 1000 tmol/L) was added to cell culture media. The proliferations of CD4+ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups (P＜0. 01 ). SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood (P＜0.01). The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4+ T lymphocytes of human periphery blood (rs = - 0. 969, P = 0. 000). Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood probably by decreasing protein levels of NF-AT to inhibit the activity and proliferation of CD4+ T lymphocytes.

Objective To evaluate the clinical value of multiplex ligation-dependent probe amplification (MLPA) technique used in karyotype analysis of chorionic villi from missed abortion. Methods Feb 2008 to Oct 2008, 91 patients with missed abortion diagnosed by hormonal measurement, type B ultrasound and physical exam matched with 20 normal pregnant women undergoing artificial abortion were enrolled in this study. Chorionic villi was obtained by suction dilation and curettage in aseptic condition, then those villi was cultured and analyzed by traditional cytogenetic karyotyping method, in the mean time, the DNA extracted from villi was detected by MLPA. The results of chromosomal G-banding of chorionic villi were compared between two methods. Results The diagnostic concordance of MLPA and traditional karyotyping was observed in 92% (84/91) cases, there were 84 cases in the case group with diagnostic concordance by traditional karyotyping and MLPA except 7 cases of euploidy could not be detected by MLPA. The 84 cases included 40 normal karyotype,29 trisomy of euchromosome, 1 double trisomy of euchromosome, 10 monosomy X , 1 monosomy X combined with trisomy of euchromosome, 2 chimaera of X chromosome, 1 structural abnormity of euchromosome. Among 7 cases with discordance diagnosis, 2 cases with trisomy and 5 cases with tetrasomy of euchromosome were identified in traditional karyotyping, however, they were all diagnosed with normal disomy by MLPA. Of 20 villi from normal pregnancy, two methods got the consistent results. Conclusion The MLPA was rapid and efficacy method used for analyzing aneuploids in chorionic villi.

Objective To analyze karyotype,genetic characteristics and clinical features of Turner syn-drome.Methods Cytogenetic analysis and genetic counseling were performed for patients with the diagnosis of Turner syndrome.Results Analyzed karyotype of forty-nine patients with Turner syndrome,there are five major categories such as simple type,numerical abnormality and chimeras,structural abnormality and chime-ras,both numerical abnormality and structural abnormality chimeras,contained the Y chromosome.Abnormal karyotypes were presented at eighteen species.Simple type was the most prevalent type of patients with Turn-er syndrome(43%).Conclusion Different karyotype exhibit some different phenotype.The clinical manifesta-tion of chimeras might depend on the proportion of abnormal karyotype,Case contained the Y chromosome would be hermaphrodismy mostly.Timely diagnosis of the disease would have positive significance to preven-tion and therapy.Hormonal therapy could improve patient's height and gonad development,the case contained the Y chromosome should pay attention to prevent gonadoblastoma.

Objective: To explore the safety and strategy of thoracic endovascular aortic repair (TEVAR) combining coronary artery bypass grafting (CABG) as one-stop performance in treating the patients with coronary artery disease (CAD) and thoracic aorta disease. Methods: A total of 20 patients received one-stop treatment of TEVAR combining CABG in our hospital from 2009-04 to 2016-01 were retrospectively analyzed. There were 18 male and the mean age of patients was (65.2±8.5, 51-82) years. The performance strategy and peri-operative management were studied. Results: There were 1/20 patient received 2 stents implantation in thoracic aorta and 19 received 1 stent in thoracic aorta those including 1 case with endovascular repair of abdominal aortic aneurysm, 1 with right iliac artery stent implantation and 1 with carotid endarterectomy at meanwhile. The average number of coronary artery bypass branch was (2.4±0.94, 1-4) and 10 (50%) patients received internal mammary artery grafting. The average in-hospital time in all 20 patients was (22.4±11.6, 8-58) days. There were 6 (30%) patients received blood transfusion; 1 (5%) having low cardiac output syndrome received extracorporeal membrane oxygenation (ECMO), then received the second thoracotomy for hemostasis due to excessive pleural effusion; 2 (10%) patients died at 30 days post-operation. 1 patient lost contact and 17 received clinical or telephone follow-up visit at the average of (13.4+13.6, 1-49) months; 2 patients died for cerebral hemorrhage at 12 and 49 months post-operation, the rest 15 had disappeared symptoms and improved quality of life, no operation related death occurred. Conclusion: TEVAR combining CABG as one-stop performance presented good mid-term effect in treating the patients with CAD and thoracic aorta disease; in otherwise, the operative time and risk might be increased by two step performance.

Objective To analyze the effects of health risk factors on presenteeism in enterprise employees.Methods A hospital in Jinan of Shandong province and a publishing enterprise in Beijing were selected as target settings of this cross-sectional study. Questionnaire survey was carried out to collect information on demographic data,health condition and presenteeism,and multiple logistic regression was used to analyze the influence of health risk factors on labor-related presenteeism.Results Insufficient physical activity was found in 42％ percent of the participants,and those with heavy-loaded pressure,overweight or obesity,high blood pressure and unhealthy diet habit accounted for 46％,41％,40％ and 34％,respectively. Proportions of employees with presenteeism varied with age ( x2 =10.1665,P =0.0377),occupation ( x2 =35.3579,P ＜ 0.05 ) and education level ( x2 =50.5377,P ＜ 0.05 ).Female employees ( P =0.0323,OR =1.728,95％ CI 1.047 to 2.850),graduates ( P ＜ 0.05,OR =6.159,95％ CI 2.510 to 15.109),overweight or obesity individuals ( P =0.0076,OR =1.673,95％ CI 1.147 to 2.44) and adults with poor self-perception of health ( P =0.0049,OR =2.284,95％ CI 1.285 to 4.060)seemed to be more likely to be presenteeism.Conclusions Overweight or obesity and poor self-perception of health may be associated with labor-related presenteeism.

OBJECTIVETo investigate a patient featuring a complex neuromuscular disease phenotype.

METHODSA comprehensive analysis integrating clinical investigation, electrophysiological testing, pathological analysis and mutation screening was carried out.

RESULTSThe patient has presented clinical and pathological manifestations mimicking Duchenne muscular dystrophy. However, genetic analysis has identified no deletion in 21 exons of Dystrophin gene, no pathologic expansion of CTG repeats in DMPK gene or CCTG repeats in ZFN9 gene. Instead, a homozygous deletion of exons 7 and 8 in SMN gene was discovered.

CONCLUSIONA rare case of spinal muscular atrophy (SMA) was verified by genetic diagnosis. SMA is a group of neuromuscular disorders with great phenotypic heterogeneity and sometimes cannot be diagnosed by clinical manifestations, electrophysiological and pathological changes alone. Genetic diagnosis has become indispensable for accurate diagnosis for patients suspected to have the disease.

Adult ; Diagnosis, Differential ; Humans ; Male ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; pathology ; Myotonic Dystrophy ; diagnosis ; genetics ; pathology ; Myotonin-Protein Kinase ; Phenotype ; Protein-Serine-Threonine Kinases ; genetics ; SMN Complex Proteins ; genetics ; Young Adult

OBJECTIVETo investigate the association between the expression of galectin- 3 protein and peritoneal metastasis in gastric cancer.

METHODThe expressions of galectin- 3 was detected in matching- samples including primary gastric cancer lesions,lymph node metastases,peritoneal metastases and paratumor normal tissues by immunohistochemistry. All specimens were gained from 35 patients who had synchronous peritoneal metastasis from gastric cancer.

RESULTSThe over- expression of galectin- 3 was observed in 97% (34/35) of the gastric cancer lesions, the peritoneal metastases and the lymph node metastases,whereas in 14% (5/35) of paratumor normal tissues. There were significant differences in the expression of galectin- 3 between paratumor normal tissues and the gastric carcinoma lesions,peritoneal metastases and lymph node metastases (P< 0.05),but there were no significant differences among the gastric cancer lesions,the peritoneal metastases,and the lymph node metastases (P> 0.05).

CONCLUSIONThe expression of galectin- 3 in gastric cancer lesions can be used as a biological marker of peritoneal metastasis from gastric cancer before operation and as a prognostic factor of gastric cancer.

Galectin 3 ; metabolism ; Humans ; Immunohistochemistry ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Neoplasm Staging ; Peritoneal Neoplasms ; metabolism ; pathology ; secondary ; Stomach Neoplasms ; metabolism ; pathology

AIMTo compare sonoporation effect of two phospholipids-based vectors-liposomes and microbubbles on cultured cell membrane.

METHODSA breast cancer cell line SK-BR-3 was exposed to ultrasound alone, 2% or 5% liposome + ultrasound and 2% or 5% microbubble + ultrasound, separately. Immediately after the experiment and 24 h after ultrasound exposure, atomic-force microscopy (AFM) scanning was used to observe the membrane change of SK-BR-3 cells.

RESULTSAfter ultrasound exposure, normal SK-BR-3 cells more or less lost their natural shape, showing elliptic outline with obtuse curved boundary. In groups added with phospholipids-based microbubbles, more obtuse curved boundary of cells was observed. The membrane pores of SK-BR-3 cells had apparent changes after ultrasound exposure. With AFM technique, membrane pores under ultrasound alone or ultrasound with liposomes conditions were enlarged, the diameter of some pores exceeding 1 microm. But all the membrane pores in these conditions returned to normal appearance after 24 hours. In ultrasound with 2% microbubble condition, most membrane pores were about 1 - 3 microm in size and returned to normal appearance after 24 h. In ultrasound with 5% microbubble condition, however, pores of most cell membrane porosity was about 2 - 4 pm and did not totally return to normal appearance after 24 h.

CONCLUSIONAt 2% concentration, phospholipids-based microbubble could enhance ultrasonic sonoporation effect and produce reparable membrane pores on SK-BR-3 cells, which appeared to be a promising vehicle for drug and gene delivery.

Cell Membrane Permeability ; Drug Carriers ; Liposomes ; Microbubbles ; Phospholipids ; chemistry ; Porosity ; Sonication ; instrumentation ; Technology, Pharmaceutical

AIMTo investigate the feasibility of transfer antisense oligodeoxynucleotides (AS-ODNs) by the phospholipids-based gas-filled microbubbles (PGM) under ultrasound activation.

METHODSAn antisense oligodeoxynucleotides sequence ZL combined with luciferase reporter plasmid was used. A breast cancer cell line SK-BR-3 was exposed to different conditions to investigate the effects of such factors as ZL concentration, PGM concentration, mechanical index (MI) and ultrasound exposure duration on transfection efficiency and cell viability. The transfection efficiency and cell viability by other lipid vectors such as lipofectamine and liposome were also tested, whose results were comparied with that of PGM. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by PI (propidium iodide) assay.

RESULTSAmong the factors tested, ultrasound exposure duration, MI and PGM concentration had obvious impacts on transfection efficiency and cell viability. The results showed that the optimal ultrasound condition was the exposure to ultrasound at MI 1.0 for 30 s with 2% PGM concentration, which gave an overall transfection efficiency of 78% +/- 10%, increased nearly 18 folds over the transfection by PGM (4.0%) or lipofectamine (4.3%) without ultrasound. Under same ultrasound conditions, different vectors showed significant difference in transfection efficiency while there are similar results in cell viability.

CONCLUSIONUnder proper ultrasound conditions, PGM can markedly enhance AS-ODNs transfection efficiency.

Breast Neoplasms ; genetics ; pathology ; Cell Line, Tumor ; Cell Survival ; Drug Carriers ; Female ; Humans ; Lipids ; Liposomes ; Luciferases ; genetics ; metabolism ; Microbubbles ; Microscopy, Fluorescence ; Oligodeoxyribonucleotides, Antisense ; genetics ; Phospholipids ; Transfection ; methods ; Ultrasonics

OBJECTIVETo analyze the drift of the complementarity-determining region 3 (CDR3) of T cell receptor beta (TCRbeta) chain variable region in T cells of healthy volunteers cultured with interleukin-2 (IL-2).

METHODST cells were isolated from the peripheral blood and cultured in vitro in the presence of IL-2. The non-specific killing effect of the cells was analyzed by LDH releasing assay, and the distribution of TCRbeta chain CDR3 in healthy volunteers by immunoscope spectratyping method to evaluate the clonality of the T cells.

RESULTSThe results showed Gaussian distribution of TCR Vbeta gene CDR3 in healthy volunteers. The T cell cultured with IL-2, however, displayed some anomalous and oligoclonal expansion in different TCR Vbeta families without killing effect against nasophargngal carcinoma cell line CNE2.

CONCLUSIONIL-2 may affect TCRbeta chain CDR3 distribution in T cells cultured in vitro.

Cells, Cultured ; Complementarity Determining Regions ; genetics ; Genetic Drift ; Humans ; Interleukin-2 ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; T-Lymphocytes ; immunology ; metabolism