Background and purpose:CD4+CD25+ regulatory T (Treg) cells are an important naturally accruing subpopulation of regulatory T cells. They not only prevent the spontaneous emergence of autoimmune diseases and contribute to the establishment of dominant tolerance on allogeneic transplantation, but also play an essential role in the generation, development and immunotherapy of many different kinds of malignant tumors. We studied the expression of CD4+CD25+ regulatory T cells(Tregs) in colorectal cancer patients and its significance. Methods:The tissues of tumors and counterpart normal tissue of 30 colorectal cancer patients were collected, the proportion of CD4+CD25+ Tregs as the percentage of the total CD4+ T cells were analyzed using flow cytometry and immunohistochemistry.Results:Compared with counterpart normal tissue(5.5?1.3)%, the percentage of CD4+CD25+ Tregs in tumor tissues was significantly higher (24.1?4.8)%(t=5.155,P=0.002). The proportion of CD4+CD25+ Tregs in the patients with lymph nodes metastasis was (27.9?3.6)% which was significantly higher than that in patients without lymph nodes metastasis[(20.3?1.3)% (t=3.489,P=0.025)].Conclusions:The proportion of CD4+CD25+ Tregs in tumor tissues of colorectal cancer patients was significantly increased, and they may play an essential role in the development of colorectal cancer.

Objective To explore and evaluate the efficiency of the leucocyte depletion filter on eliminating tumor cells from blood, and its influence on the morphology and function of red blood cells. Methods Blood from operation field was collected, filtered, centrifuged and washed. Intestinal cancer cells (LOVO) and gastric cancer cells (SGC) were cultured in vitro and mixed with the collected blood. Blood contaminated with known number of tumor cells was then centrifuged and washed with cell retriever, and then the mixture was filtered with leucocyte depletion filter. The amount of residual tumor cells in filtered blood was counted after trypan blue staining before and after culturing. Unwashed and unfiltered blood containing tumor cells were used as control and were also cultured. The morphology and Na+-K+-ATPase activity of treated red blood cells was assessed. Results Tumor cells were detectable after only being washed with cell retriever and grew well after culturing for two weeks. No tumor cells were found after both washed with cell retriever and filtered with leucocyte depletion filter, and no tumor cell grew two weeks after culturing. No change was found in morphology and Na+-K+-ATPase activity of red blood cells after being washed and filtered. Conclusion Cell retriever is not able to eliminate tumor cells from blood completely. Further filtration with leucocyte depletion filter is capable of completely eliminating the residual tumor cells in blood. The morphology and Na+-K+-ATPase of red blood cells do not change after being washed with cell retriever and being filtered with leucocyte depletion filter.

Objective To investigate the incidence of glucose metabolism disorder during pregnancy and establish the diagnosis criteria for gestational diabetes mellitus (GDM) among Chinese patients.Method A prospective popolation-bused study of 16 286 pregnant women,who received 50 g glucose challenge test (GCT) for the first time between Apr 1,2006 and Sept 30,2006,was performed throughout 18 cities in China. Results According to national diabetes data group (NDDG)criteria,the overall incidence of GDM and glucose impaired glucose tolerance (GIGT) was 2.763% (450/16 286) and 3.862% (629/16 286),respectively; it changed to 5.078% (827/16 286)and 5.268% (858/16 286) when using American diabetes association (ADA) criteria.If the women who had 2 or more abnormal oral glucose tolerance test (OGTT) values meeting or exceeding ADA criteria was classified as group 1,and the women who had one or more meeting or exceeding NDDG criteria was group 2,the ratio of women who met both the criteria of ADA and NDDG in group 1 was 94.2%.The 95% CI of normal glucose when using ADA criteria were fasting glucose 5.3 mmol/L,1 hour 10.4 mmol/L,2 hour 8.7 mmol/L,3 hour 7.7 mmol/L,which is close to ADA criteria.Conclusions ADA criterion is more suitable for Chinese patients.According to NDDG criteria,it is reasonable to treat the patient with 1 or more abnormal OGTT values,and if choosing ADA criteria,2 or more abnormal OGrIT values is more reasonable.

Objective To evaluate the effect of pretreatment with butorphanol to prevent injection pain of rocuronium bromide.Methods One hundred and fifty ASA Ⅰ-Ⅱ grade patients,undergoing elective surgery were divided into 3 groups by random digits table with 50 cases each.Butorphanol group received butorphanol 2 mg,fentanyl group received fentanyl 100 μg,and control group received 0.9％sodium chloride when general anesthesia induced.Anesthesia was induced with propofol 2 mg/kg and the test drug was injected over 30 s,120 s after the test drug injection,1％ rocuronium bromide 0.6 mg/kg was injected.Nausea,vomiting,apnea and bucking were recorded after drugs injection.Results The rates of rocuronium bromide injection pain in fentanyl group[10％(5/50)]and butorphanol group[8％(4/50)]were significant lower than that in control group[82％(41/50)](P ＜ 0.01).None of the patients discovered nausea,vomiting,apnea and bucking in 3 groups.Conclusion Pretreatment with 2 mg butorphanol reduced the incidence of rocuronium bromide injection pain,furthermore,there is no untoward reaction such as nausea,vomiting,apnea and bucking.

Objective To localize the upper airway obstruction of patients with obstructive sleep apnea hypopnea syndrome (OSAHS)with the Cinema Magnetic Resonance(Cine-MR) and fiber optic laryngoscope with Müller maneuver(FLMM)before operation and discuss the clinical application values.Methods Before operation, FLMM and Cine-MR were applied to 22 patients diagnosed as OSAHS by Polysomnography(PSG).Medical examinations conducted in this study from September 2015 to April 2016 to examine the obstruction of the soft palate region, the lingual region and epiglottis.Results There were complete agreements between the Cine-MR and FLMM at locating obstruction sites of the soft palate (n=22/n=22),and there were moderate agreements between the Cine-MR and FLMM in locating obstruction sites of the retroglottal region(n=13/n=6),epiglottal region (n=4/n=2)and multiple level(n=13/n=6), respectively.Conclusion For those moderate and severe OSAHS patients with multiple sites obstruction , the preoperative application of the Cine-MR and FLMM together will be better in locating the obstruction sites.

ObjectiveTo evaluate the accuracy of cerebral state index (CSI) as an indicator of anesthesia depth in the induction of anesthesia with target-controlled infusion (TCI) with propofol and remifentanil patients.Methods Forty-four ASA Ⅰ or Ⅱ patients undergoing elective surgery were anesthetized with TCI with propofol and remifentanil.Anesthesia was induced with TCI with remifentanil and propofol.The target effect-site concentration of remifentanil was 4 ng/ml.The initial effect-site concentration of propofol was 1.5 μ g/ml and was increased by0.5 μ g/ml every 4 min,till 1 min after the level of observer’s assessment of alertness sedation (OAA/S) score was 0 score.Electric tetanic stimulation was given when the level of OAA/S score was 1 score.The CSI,mean arterial pressure (MAP),heart rate (HR),OAA/S score and the effect-site concentration of propofol were recorded.ResultsCSI values declined with the decrease of OAA/S score,CSI was 91 ±5,77 ±7,70 ±7,62 ±6,49 ± 12,36 ± 10 at OAA/S score with 5,4,3,2,1score.CSI values were statistically different between 0 score and 1 score,1 score and 2 scores,3 scores and 4 scores,4 scores and 5 scores of OAA/S score(P ＜0.05).The differences of MAP,HR had no statistical significance between two scores of OAA/S score (P ＞0.05).The Spearman rank correlation coefficients between CSI,MAP,HR and OAA/S score were 0.899,0.342,0.125,respectively.The prediction probabilities to differentiate different OAA/S score for CSI,MAP,and HR were 0.89 ± 0.05,0.62 ± 0.08,0.53 ±:0.11,respectively.There was linear regression relationship between CSI and the effect-site concentration of propofol (the coefficient of determination was 0.812,P ＜ 0.01 ).ConclusionDuring the induction of patients with TCI with propofol and remifentanil,the CSI is accurate as an indicator of awakening and different levels of consciousness after anesthesia,and can reliably predict the anesthesia depth.

Objective To investigate the regulation of the immune function of cytokine-induced killer cells(CIK) by dendritic cell (DC) vaccine in the patients with chronic hepatitis B(CHB) in vitro.Methods CIK cells from 30 patients with CHB were cultured in vitro,and were randomly divided into two groups,DC vaccine-treated group and the control group.After 14 days of culture,the percentages of CD3 + CD4+ T,CD3 +CD8 +T and CD3 +CD56+ T cells among CIKs were analyzed by flow cytometry.The levels of IL-12,IFN-γand IL-6 in cell culture supernatant was detected by ELISA.Results The percentages of CD3 + CD4 + T,CD3 +CD8+T and CD3+ CD56+ T cells were 18.27％,64.36％ and 20.00％ in CIKs in DC vaccine group,and 17.79％ ( P ＞ 0.05 ),54.69％ ( P ＜ 0.01 ) and 13.39％ ( P ＜ 0.01 ) in the control group,respectively.The perentage of CD3 + CD4 + T cells were similar between the two groups ( P ＞ 0.05 ),but for the perentage of CD3 + CD8 +T and C D3 + CD56 + T cells were significantly different between the two groups (t =4.130 and 5.601respectively,Ps ＜ 0.01 ).The concentrations of IL- 12,IL-4 and IFN-γin supernatant were ( 177.82 ± 130.06),(31.77 ± 9.52) and (86.99 ± 56.30) ng/L in DC vaccine-treated group respectively,which were significantly different from those of (80.83 ±50.15) ng/L (t =3.811,P ＜0.01 ),(40.33 ± 19.74) ng/L( t =2.141,P ＜0.05) and (42.07 ± 19.68)ng/L(t =4.125,P ＜0.01) in the control group,respectively.Conclusion DC vaccine could enhance the killing function of CIK cells.

BACKGROUND:At present, the incidence rates of knee joint diseases such as knee osteoarthritis, knee joint degenerative are high. The major clinical treatment is total knee replacement in the clinic, so it is necessary to evaluate the changes in stress and bone mineral density of the regions surrounding the prosthesis after replacement. ＜br> OBJECTIVE:To explore periprosthetic stress and bone mineral density and to analyze their correlation after total knee arthroplasty. ＜br> METHODS:A total of 20 cases undergoing total knee arthroplasty were chosen.The hospital for special surgery scores were used to evaluate patients’ functional recovery at 12 months after total knee arthroplasty. The periprosthetic femur was divided into four regions of interest (ROI), respectively ROI 1-4;periprosthetic tibia was divided into three regions of interest, respectively ROI 5-7. Stress surrounding the prosthesis was analyzed using three-dimensional finite element analysis at 1, 3, 6 months, 1, 2, 3 years after replacement. Simultaneously, bone mineral density surrounding the prosthesis was measured using dual-energy X-ray absorptiometry. ＜br> RESULTS AND CONCLUSION:No patients affected infection or loosening of the prosthesis. At 12 months after replacement, the score of hospital for special surgery was (90.23±2.37), which showed significant differences as compared with before replacement (39.68±1.31) (P＜0.05). The level of stress shielding was highest in ROI 5 and lowest in ROI 3. Stress shielding rate of ROI increased with statistical difference at 6 months after operation (P＜0.05). At 1, 2, 3 years after operation, shielding rate in periprosthetic femoral stress in ROI 1 decreased. Compared with 1 month after operation, the difference was statistical y significant (P＜0.05). However, shielding rate of tibial periprosthetic stress in ROI 6 increased. Compared with 1 month after operation, the difference was statistical y significant (P＜0.05). Bone mineral density after 1 month after operation had no significant decrease (P>0.05). At 3 months after operation, bone mineral density began to decline significantly (P＜0.01). The decrease was most obviously in ROI 5 and the change was least in ROI 3. After 1 year of operation, bone mineral density did not change significantly. These data indicated that changes in bone mineral density were correlated with stress shielding after total knee arthroplasty. Monitoring two variations can provide theoretical data for preventing bone loss, which provides references for clinical rehabilitation guidance.

Objective To compare SPECT/CT fusion imaging and MRI in the diagnosis of benign hip lesions. Methods Twenty-two patients suspected avascular necrosis of femoral head with hip discomfort, pain or action limited were analyzed retrospectively. All patients underwent radionuclide bone scan and MR examination within 5 days, and the diagnosis was proved with clinical follow-up. Results Eighteen necrosis of the femoral head and 4 hip arthritis including 1 patient with ankylosing spondylitis were found in 44 hip joints of 22 patients. MRI detected 17 femoral head necrosis and 4 hip arthritis, while SPECT/CT fusion image found out 18 femoral head necrosis and 4 hip arthritis. There was corresponding relationship in signs of hip lesions between MRI and SPECT/CT fusion imaging. Conclusion SPECT/CT fusion imaging and MRI has no markedly difference in the diagnosis of hip benign lesions, and is complementary to each other. SPECT/CT fusion image can distinguish the hip lesions from the femoral head lesions, and has a higher accuracy of diagnosing hip lesions than whole body bone scanning.

Objective To understand the mutation characteristics of drug resistance-associated genes rpoB, katG and inhA in Mycobacterium tuberculosis (MTB) strains using gene chip method, and evaluate its clinical application value. Methods A total of 76 MTB strains were collected from Shijiazhuang area in 2013 to 2014. Gene chip was used to detect the mutations of rpoB, katG and inhA, and the L-J proportion drug susceptibility test was used as the gold standard to evaluate the overall concordance, sensitivity and specificity of gene chip. The consistency of microarray and phenotypic resistance was evaluated by Kappa test. Results Of all the 76 strains detected, 69 harbored mutations in katG/inhA. The predominant mutation site of katG was 315 codon with the mutation rate of 89.9%(62/69), and 5.8%(4/69) carried mutations at inhA-15(C→T), and 4.3%(3/69)carried combined mutations of katG 315 and inhA-15. The rpoB mutations were detected in 73 strains, of which 64.4%(47/73)carried mutations at codon 531, 15.1%(11/73)at codon 526, 12.3%(9/73)at 516 codon, 1.4%(1/73)at 513 codon, 1.4%(1/73)at 533 codon and 5.5%(4/73)had combined mutations. Compared with results from the L-J proportion method, the sensitivity, specificity and concordance rates of gene chip for RFP were 96.1%(73/76), 100%(50/50)and 97.6%(123/126). The sensitivity, specificity and concordance rates of gene chip for INH were 90.8%(69/76), 100%(50/50)and 94.4%(119/126). The sensitivity, specificity and concordance rates of gene chip for MDR-TB were 86.8%(66/76), 100%(50/50) and 92.1%(116/126). Conclusion The predominant mutation loci of MDR strains in Shijiazhuang area are katG315 and rpoB531. Gene chip is a fast and useful tool for clinical diagnosis of MDR strains.