Objective To investigate the phenotype and the immunoregulatory function of CD8αα+TCRαβ+regulatory T cells in peripheral blood samples from mice.Methods The distribution profile and the phenotype of CD8αα+TCRαβ+regulatory T cells in C57BL/6 mice were detected by flow cytometry.The cytokines released by CD8αα+TCRαβ+regulatory T cells upon the stimulation with anti-CD3 antibody were analyzed by cytometric bead array.The in vitro immunosuppressive activity of CD8αα+TCRαβ+regulatory T cells on activated CD4+T cells was analyzed by using flow cytometry and carboxyfluorescein succinimidyl ester ( CFSE ) .An adoptive cell transfer assay was set up to evaluate the immunoprotective effects of CD8αα+TCRαβ+ regulatory T cells in a mouse model of experimental autoimmune encephalomyelitis ( EAE) .Results CD8αα+TCRαβ+regulatory T cells were detected in liver, spleen and peripheral blood samples collected from na?ve C57BL/6 mice.Compared with CD8αβ+TCRαβ+regulatory T cells, CD8αα+TCRαβ+regulatory T cells showed a memory-activated phenotype of CD25+CD122high CD44high CD62Llow CD69high NK1.1+DX5+.CD8αα+TCRαβ+regulatory T cells could produce IL-2 after 24 hours stimulation with anti-CD3 antibody, followed by producing IFN-γ, TNF-α, IL-4, IL-17A and traces of IL-6 and IL-10. In vitro, CD8αα+TCRαβ+regulatory T cells specifically suppressed the proliferation of activated CD4+T cells ( P<0.01 ).Moreover, they could delay the onset of EAE in mice and reduce clinical score (P<0.01).Conclusion CD8αα+TCRαβ+regulatory T cells were a unique population with immunoregula-tory function, which could be used as a potential therapeutic target in the treatment of autoimmune disease.

A method was proposed for the separation and determination of paraffin waxes in food by HPLC-evaporative light scattering detection (ELSD). A normal-phase column was used to separate nonparaffinic and paraffinic materials without resolving the latter into individual components. The t-test method was adopted for the evaluation of mean difference between response factors of n-alkanes in paraffin waxes on ELSD detector. No mean difference was obtained between response factors, which can be used for quantitative determination of paraffin waxes in food. The determination results obtained by HPLC-ELSD were compared with those by GC-MS. The linear range for the determination of paraffin waxes was in the range from 10 to 500 mg/L with a correlation coefficient of 0.9988, and the limit of detection was 1.0 mg/L. With the spiking level of 10, 50 and 100 mg/kg, the recovery ranged from 84.6% to 105.4% and the relative standard deviation ranging from 5.4% to 7.2%. The proposed method is simple, fast and sensitive.

OBJECTIVETo evaluate the clinical results after arthroscopic rotator cuff repair using a suture-bridge technique in late middle-aged adults.

METHODSA retrospective analysis of 30 patients with rotator cuff tears, who were treated consecutively between December 2012 and December 2013 at our clinic, was carried out to evaluate the outcome of arthroscopic suture-bridge technique. The visual analogue scale (VAS), the range of motion (ROM), the rating scale of the University of California at Los Angeles (UCLA), and the shoulder index of the American Shoulder and Elbow Surgeons (ASES) were determined as the evaluation criteria.

RESULTSThe mean follow-up period was 13 months. The follow-up rate was 100%. The study included 0 small, 16 medium, 12 large and 2 massive rotator cuff tears. At the latest follow-up, the VAS score decreased significantly from preoperative 6.3±3.2 to 0.8±1.4 at rest,and from preoperative 7.7±2.1 to 1.7±1.6 during motion. The UCLA score improved significantly from 15.1±4.6 preoperatively to 31.2±5.0 at follow-up. The ASES shoulder index improved significantly from 30.2±16.0 preoperatively to 80.4±13.0 at follow-up. The ROM improved significantly. The satisfactory rate was 94%. There were no complications.

CONCLUSIONThe results of this study indicate that the arthroscopic suture-bridge technique is an effective and reliable treatment for patients with rotator cuff tear in late middle-aged adults.

Age Factors ; Aged ; Arthroscopy ; methods ; Female ; Humans ; Male ; Middle Aged ; Range of Motion, Articular ; Rotator Cuff ; surgery ; Suture Techniques

This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg kg(-1) dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg kg(-1) dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.
Animals ; Apoptosis ; drug effects ; Asteraceae ; chemistry ; Brain Ischemia ; metabolism ; pathology ; Cerebral Cortex ; metabolism ; pathology ; Dose-Response Relationship, Drug ; Glucosides ; administration & dosage ; isolation & purification ; pharmacology ; Male ; Neurons ; drug effects ; pathology ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; Rhizome ; chemistry ; Vasodilator Agents ; administration & dosage ; isolation & purification ; pharmacology ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo find out the mechanisms of redifferentiation and reversion of malignant human gastric cancer cells induced by ascorbic acid.

METHODSHuman gastric cancer cells grown in the laboratory were used. The Trypan blue dye exclusion method was used to determine the cell doubling time. The electrophoresis rate and colonogenic potential were the indices used to measure the rate of redifferentiation. The content of malondialdehyde (MDA) was measured using the thiobarbituric acid (TBA) method. The activities of superoxide dismutase (SOD), catalase (CAT) and the content of H202 were evaluated by spectrophotography.

RESULTSSix mmol/L ascorbic acid was used as a positive control. Human gastric cancer cells were treated with 75 microm hydrogen peroxide, which alleviated many of the malignant characteristics. For example, the cell surface charge obviously decreased and the electrophoresis rate dropped from 2.21 to 1.10 microm x s(-1) x V(-1) x cm(-1). The colonogenic potential, a measure of cell differentiation, decreased 90.2%. After treatment with ascorbic acid, there was a concentration- and time-dependent increase in hydrogen peroxide (H202) and the activity of superoxide dismutase (SOD). However, the activity of catalase (CAT) resulted in a concentration- and time-dependent decrease. SOD and 3-amino-1,2,4-triazole (AT) exhibited some effects, but there were statistically significant differences between the SOD and AT group and the H202 group.

CONCLUSIONSAscorbic acid induces growth inhibition and redifferentiation of human gastric cancer cells through the production of hydrogen peroxide.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cell Differentiation ; drug effects ; Humans ; Hydrogen Peroxide ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured

Objective To evaluate the feasibility and safety in the treatment of liver cancer located under the diaphragm with cool-tip radiofrequency ablation(RFA)percutaneously under CT guidance.Methods 20 patients with total 25 lesions were treated by CT-guided RFA with cool-tip electrode involving the induced necroses.The postoperative efficacy was evaluated by enhanced CT or MRI.Results 72% lesions were completely necrotized(18/25),28% lesions were majorly necrotized(7/25).No severe complications occurred. Conclusion CT-RFA with cool-tip electrode is effective and safe in treating liver cancer located under the diaphragm.

Objective:To observe the effects of electroacupuncture (EA) of different frequencies on the expression levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the colon of rats with slow transit constipation (STC).Methods:One hundred healthy male Sprague-Dawley (SD) rats were randomly divided into a normal group,a model group,a low-frequency EA group,a high-frequency EA group and a variable-frequency EA group,with 20 rats in each group.The rats in the normal group were fed with normal diet.The rats in the other groups were fed with phenethylpiperidine in the diet at a dose of 8 mg/(kg·bw) per day,for 120 d,to establish the STC model.Rats in the normal group and the model group did not receive any treatment;rats in the low-frequency EA group were treated with 2 Hz continuous wave EA,rats in the high-frequency EA group were treated with 100 Hz continuous wave EA,and rats in the variable-frequency EA group were treated with 2 Hz/100 Hz sparse-dense EA.The current intensity of the EA was determined by the slight vibration of the rat limbs without painful screaming.The intervention was performed once a day,15 min/time for continuous 15 d.After treatment,the intestinal transit function and the expression levels of SP and VIP in the colon of the rats in each group were determined.Results:After treatment,the defecation duration of the first dark stool in the model group was significantly longer than that in the normal group (P＜0.05);the defecation durations of the first dark stool in the low-frequency EA group,high-frequency EA group and variable-frequency EA group were significantly shorter than the duration in the model group (all P＜0.05);compared with the low-frequency EA group,the first dark stool defecation duration of rats in the variable-frequency EA group was significantly shorter (P＜0.05);compared with the normal group,the SP and VIP expression levels in the colon of the model group were significantly decreased (both P＜0.01);the SP and VIP expression levels in the colon of the low-frequency EA group,the high-frequency EA group and the variable-frequency EA group were significantly higher than those in the model group (all P＜0.05);compared with the high-frequency EA group,the SP expression levels in the colon in the low-frequency EA group and the variable-frequency EA group were significantly increased (both P＜0.05);compared with the low-frequency EA group,the VIP expression levels in the colon in the high-frequency EA group and the variable-frequency EA group were significantly increased (both P＜0.05).Conclusion:EA improves the intestinal function of STC model rats by regulating the expression levels of SP and VIP in rat colon.The EA stimulation with 100 Hz continuous wave,2 Hz/100 Hz sparse-dense wave shows a better improvement in the colonic transit function in STC rats,followed by 2 Hz continuous wave.

Objective To explore the current status of the prenatal care quality and the influencing factors in Hangzhou City. Methods We used sociodemographic characteristics questionnaire and the Chinese version of QPCQ to investigate 383 pregnancy women who were at 36 weeks' gestation or greater by convenient sampling method from September to December in 2016. Results A total of 350 questionnaires were valid. The average age of 350 samples was (29.71±3.67) years old. There were 231 primiparas which accounted for 66.00％. Among them, 196 (56.00％) were satisfied with the prenatal care they received, and 153 (43.70％) think general, and only 1 (0.30％) felt unsatisfied. The score of the Chinese version of QPCQ was 3.61±0.50. The item "My prenatal care professional (s) always answered my questions honestly" had the highest scores of 4.10±0.62, while "My prenatal care professional (s) did things hastily" had the lowest scores of 2.44±0.96 . By multiple linear regression analysis, those people who had more visiting time (β' = 0.21, P<0.01), less waiting time (β' = -0.18, P<0.01) and the cognition of the essentiality of pregnancy school (β' = 0.12, P=0.02) were more likely to give higher regards for the quality of prenatal care. Conclusion Sufficient time scored relative lower in the whole questionnaire. So, hospitals could take efforts to shorten the waiting time by promoting the hospital process optimization, making good use of this period time and other strategies, so as to improve the quality of prenatal care.

OBJECTIVETo investigate the relationship of erectile dysfunction (ED) with blood vessel-, nerve- and androgen-related factors in young and middle-aged men with type 2 diabetes mellitus (T2DM) in order to provide some clinical evidence for early prevention and treatment of ED.

METHODSWe divided 53 male T2DM patients under 50 years into an ED group (IIEF-5 score < or = 21, n = 28) and a non-ED (NED) group (IIEF-5 score > or = 22, n = 25). We detected the levels of blood lipid, glucose, total testosterone (TT), sex hormone-binding globulin (SHBG), sulfate dehydroepiandrosterone (DHEA-S), calculated free testosterone (cFT), and examined the complications of macroangiopathy (MA), diabetic retinopathy (DR) and diabetic peripheral neuropathy (DPN), and compared the above indicators between the two groups.

RESULTSThere were no significant differences between the two groups in age, diabetes duration, body mass index, blood pressure, and blood lipid and glucose levels (P > 0.05). The incidence rate of DR was significantly higher in the ED than in the NED group (39.3% vs 4.0%, P < 0.05), but no statistically significant differences were found in the levels of TT, cFT, SHBG and DHEA-S and the incidence rates of MA and DPN between the two groups (P > 0.05).

CONCLUSIONThe incidence of ED is closely related to DR in young and middle-aged men with T2DM. Therefore particular attention should be paid to the erectile function of T2DM patients with DR as early as possible.

Adult ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Neuropathies ; complications ; Diabetic Retinopathy ; complications ; Erectile Dysfunction ; etiology ; Humans ; Male ; Middle Aged

OBJECTIVETo identify associated proteins involved in the molecular response of ischemic preconditioning (IPC) against intestinal ischemia/reperfusion (II/R) in the intestinal mucosa of rats.

METHODSSixteen SD rats were randomly divided into II/R and IPC groups. II/R injury in rats was produced by clamping superior mesenteric artery for 60 min followed by 60 min reperfusion. IPC was elicited by 20 min ischemia and 5 min reperfusion before index ischemia. The intestinal mucosa was scratched immediately after 60 min of reperfusion and total proteins were separated by immobilized pH gradient (IPG) based on two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed using Image Master 2D Elite 5.0 image analysis software and identified by MALDI-TOF-MS. The biological information of these proteins was searched in the database of these peptide mass finger-printing (PMF). Western blotting and RT-PCR were used to validate the differentially expressed proteins.

RESULTSImage analysis revealed that averages of 1404+/-20 and 1338+/-20 were detected in II/R and IPC groups. A total of 10 spots yielded good spectra, and 8 spots matched with known proteins after database searching. These proteins were mainly involved in anti-oxidation, inhibiting apoptosis and energy metabolism. Western blot confirmed up-regulation of aldehyde dehydrogenase and RT-PCR confirmed up-regulation of aldose reductase in IPC group.

CONCLUSIONThe clues provided by comparative proteome strategy will shed light on molecular mechanisms of IPC against II/R injury.

Animals ; Intestinal Diseases ; metabolism ; pathology ; Intestinal Mucosa ; metabolism ; pathology ; Ischemic Preconditioning ; Male ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology