Objective Percoll density gradient centrifugation and Ficoll-Hypaque density gradient cen-trifugation, which are frequently-used methods for separation of tumor-associated macrophages (TAMs) from solid carcinoma were compared, in order to find an effective way to separate TAMs from colorectal carcinoma (CRC). Furthermore, we studied the best adherence time of separating macrophage among mononuclear cells. Methods specimens were collected from CRC patients , after digesting into single cells , TAMs were separated from the same specimen by 100% Ficoll, 35% percoll and 25% combined with 65% percoll respectively. After these pre-liminary separation, the collected cells were purified a second time by adherence separation. The purity of TAMs were detected by immunofluorescence. Results TAMs purity from Ficoll-Hypaque density gradient centrifugation was 80.18%, statistically higher than that from Percoll density gradient centrifugations (54.33% and 10.93% re-spectively). Conclusion Compared to Percoll density gradient centrifugation, Ficoll-Hypaque density gradient centrifugation is a more effective and simple way to isolate TAMs from colorectal carcinoma , suggesting it can be wildly used in clinical and basic medical research. 2-4 hours is the best adherence time for isolating macrophage.

Objective To investigate the level of human cytomegalovirus(HCMV)specific CD8+ T lymphoeytes in peripheral blood and the immune reaponae of HCMV reactivation after kidney transplantation.Methods Thirty-eight HCMV seropesitive HLA-A*0201 kidney transplant recipients(9 with HCMV infection and 29 without HCMV infection)and 54 healthy individuals were enrolled.The levels of total HCMV specific CD8+ T cells were measured using HLA-A2 pentamer folded with HCMV-peptide NLVPMVATV.The levels of IFN-γ secreting CD8+ T cells were measured by intracelluhr IEN-γ staining pulsed with the same peptide.Results The median levels of pentamer stained CD8+ T cells were 1.19%(0-19.42%),1.20%(0-18.40%)and 3.2%(0.51%-18.90%)in healthy group,negative HCMV group and positive HCMV group(H=5.34,P＞0.05),respectively.The median levels of IFN-γ secreting CD8+ T cells were 0.72%(0-0.70%),0.47%(0-5.61%)and 0.67%(0.07%-4.00%),respectively(H=0.58,P＞0.05).However,the mean proportions of IFN-γ secreting pentamer stained T cells relative to total HCMV specifc CIL were(60.18±19.16)%,(39.19±17.22)% and(20.02±13.26)%,respectively.There were significant differences among the groups(P＜0.01).Condusiorm There was no significant difference of levels of HCMV specific CD8+ T lymphocytes in peripheral blood between the kidney transplant recipients and healthy individuals.However,the proportion of HCMV-specific IFN-γ producing CD8+ T cells in pentamer stained cells was reduced in the kidney transplant recipients especially in those with active HCMV infection,which may contribute to the inability to control HCMV reactivation.

Objective To discuss the feasibility of positioning the acetabular center,fixing acetabular implant correctly and reconstructing hip rotation center according to Harris fossa and the remaining anatomical markers of acetabular notch in revision hip arthroplasty.Methods Twenty-eight patients underwent revision hip arthroplasty from April 2007 to June 2009.Based on Paprosky type,3 cases with type Ⅰ were treated with biological fixed acetabular component;8 cases with ⅡA and ⅡB were reconstructed with using of morselized bone grafting and large diameter cemented acetabular prosthesis;17 cases with type ⅡC,ⅢA and ⅢB were treated with using of morselized bone grafting and fixation of acetabular reinforcement ring.Among them,5 patients with massive bone loss in acetabular wall were reconstructed with the use of the structural and morselized bone grafting.The center of the original acetabulum was believed to be in the lunate cartilage surface which was closed to Harris fossa.During the operation,the center was located in the site which was 25-28 mm above in line with perpendicular bisector of acetabular notch connecting line.The acetabular center was the point of positioning acetabular prosthesis (Ⅰ type) or making new acetabulum by impaction bone grafting.Acetabular reinforcement ring (Ⅱ,Ⅲ type) was fixed in accordance with proper transverse angle and anteversion angle.The vertical distance from hip rotation center to teardrop connection and the horizontal distance from hip rotation center to teardrop were measured on preoperative and postoperative radiograph.And the outcomes of reconstruction of rotation center were evaluated.Results The vertical distance was changed from (14.22±3.39) mm preoperatively to (32.64±4.51) mm postoperatively.The difference was statistically significant (t=3.65,P＜ 0.05).The horizontal distance was changed from (25.13±3.46)mm preoperatively to (32.87±4.73) mm postoperatively.The difference was statistically significant (t=2.72,P＜0.05).Conclusion Using residual Harris fossa and acetabular notch as the anatomical markers in revision hip arthroplasty,the restoration of the anatomical hip center has shown to be favorable.

Objective: To test applicability of the Center for Epidemiological Studies Depression Scale (CES-D) in different age groups in urban China, and to develop age norms. Methods: In this cross-sectional study, the CES-D was administrated to 16047 community participants with average age of (37.7±21.3) years (age ranged 11～100) in 21 provinces, who were as the norming sample. Criterion validity was tested in 349 psychiatric patients with average age of (32.0±12.1) years (age ranged 16～81) in 4 cities. A subsample (199 workers, 100 col-lege students, and 30 teachers in Beijing, Dongguan, and Baotou) was drawn from the national sample to provide 8 week interval test-retest reliability. Results: The Cronbach α was 0.90 for the scale, and 0.68～0.86 for its fac-tors. The 8 week interval test-retest correlation was 0.49 for the scale (P <0.01) and 0.39～0.51 for factors (P<0.01) . The result of confirmatory factor analysis supported the original 4-factor structure (RMSEA=0.057, CFI =0.976, GFI=0.948) . Patients scored higher than community sample [(21.72±13.39 ) vs.(13.24±10.33),P <0.01], and depression patients scored the highest [(27.82±14.42), P<0.01] . Age difference was signifi-cant. Age groups over 60-year-old scored higher than all the other age groups under 60-year-old (P<0.01). Con-clusion: The Chinese version of CES-D shows good refiability and validity across all ages in urban population-

Objective To analyze the characteristics of TCM syndromes of children abnormal sweating;To investigate the correlation between diseases and primary and secondary syndromes. Methods Through a multi-center epidemiology investigation, 400 cases were collected to statistically analyze the five TCM syndromes of children abnormal sweating, and characteristics of each syndrome were analyzed. Results In the 400 cases of children abnormal sweating, there were 129 cases (32.25%) of dysfunction of lung-wei pattern, 50 cases (12.50%) of nutrient qi and defensive qi discordance, 84 cases (21.00%) of qi and yin deficiency pattern, 57 cases (14.25%) of dampness heat forced steamed pattern, and 80 cases (20.00%) of qi and yang deficiency pattern. Spontaneous sweating and night sweating were found in the four patterns besides qi and yin deficiency pattern, and night sweating can be mainly found in qi and yin deficiency. Each type is consistent in the syndrome differentiation and cluster analysis. Conclusion Spontaneous sweating and night sweating, each has the certification of yin and yang. Spontaneous sweating can be caused by yin deficiency while night sweating can be caused by yang deficiency. Each pattern of children abnormal sweating has its own syndrome characteristic, which can provide the basis for clinical diagnosis and treatment.

Objective To investigate the metabolite changes in systemic lupus erythematosus (SLE) patients with and without neuropsychiatric symptoms using magnetic resonance spectroscopy (MRS) and explore the associations between image findings and clinical variables. Methods Twenty-two SLE patients with neuropsychiatric symptoms (NPSLE), twenty-one SLE patients without neuropsychiatric symptoms (non-NPSLE) and twenty healthy controls (HCs) underwent routine MRI scan and multivoxel magnetic reson-ance spectroscopy (MVS). The absolute metabolite concentrations were measured bilaterally in the posterior cingulate gyrus (PCG), dorsal thalamus (DT), lentiform nucleus (LN) and posterior paratrigonal white matter (PWM) using LCModel and SAGE software. The relationships between metabolite con-centrations and cognitive function scores were analyzed by Spearman rank correlation. Single-factor Chi-square analysis and t-test were used for analysis. Results ① Compared to control subjects, NPSLE patients had significantly lower N-acetylaspartate (NAA) values in bilateral PCG and DT, with the mean differences of -1.504 [95% confidence interval ( CI) (-2.335, -0.672), P=0.001], -1.460 [95%CI (-2.349, -0.570), P=0.002], -1.259 [95%CI (-1.894, -0.625), P=0.000] and -1.022[95%CI (-1.688, -0.356), P=0.003] for RPCG, LPCG, RDT and LDT, respectively. The concentration of total creatinine were observed to decline in RPCG and RDT, with the mean differences of-1.094 [95%CI (-1.845, -0.342), P=0.003], -0.955 [95%CI (-1.630, -0.280), P=0.006], -1.259 [95%CI (-1.894,-0.625), P=0.006] respectively. Glutamine and glutamate-values decreased significantly in RDT [mean difference=-2.586, 95%CI (-4.139, -1.033), P=0.002]. ② Compared to non-NPSLE patients, NPSLE patients had a lower NAA level in LPCG [mean difference=-1.256, 95%CI (-2.146, -0.367), P=0.006]. Positive correlations between mini-mental state examination scores [RPCG: rs=0.312, P<0.05; LPCG: rs=0.355, P<0.01], Montreal cognitive assessment scores (RPCG: rs=0.362, P<0.01; LPCG: rs=0.285, P<0.05) and NAA values in bilateral PCG were detected. Conclusion Both NPSLE and non-NPSLE patients may have metabolite dysfun-ctions in different brain regions. The cognitive disorder in SLE patients may be interpreted by neuronic damage of PCG.

OBJECTIVE: To investigate the diagnosis applying effects of ocular vestibular evoked myogenic potentials(oVEMP) in peripheral BPPV disease. METHOD: During September 2012 to January 2015, we selected 80 healthy people in our hospital medical center as the control group, choose the same period of primary benign paroxysmal positional vertigo as the observation group of 80 patients. Two groups were carried out fully functional auditory evoked potential analysis, determination of oVEMP and cervical vestibular evoked myogenic potentials (cVEMP) anomaly amplitude threshold, P1 latencies, N1 incubation period. RESULT: The cVEMP abnormal rate in the observation group was 28.8%, the oVEMP abnormal rate was 38.8%, while cVEMP and oVEMP abnormal rates in the control group was 1.3% and 2.5% respectively that compared to significant differences between the two groups (P < 0.05). The oVEMP test amplitude in the observation group was (5.98 ± 2.15) µv, the N1 incubation period was (10.03 ± 0.76)ms, while the control group were (4.09 ± 2.11)µv and (11.67 ± 0.78) ms that compared difference were statistically significant (P < 0.05). The cVEMP test amplitude in the observation group was (154.8 ± 43.9)2 µv, while the control group was (180.49 ± 45.34)µv, compared the difference was statistically significant (P < 0.05). CONCLUSION Paroxysmal positional vertigo patients ocular vestibular evoked myogenic potentials abnormal rate is relatively high, the utricle dysfunction for more severe than the balloon can be the subject of an objective function of the ear stone judgment, judgment in favor of the disease.
Benign Paroxysmal Positional Vertigo ; diagnosis ; Case-Control Studies ; Humans ; Saccule and Utricle ; physiopathology ; Vestibular Evoked Myogenic Potentials

Objective To evaluate the diagnostic and prognostic value of BRAF V600E mutation screening of ultrasound-guided fine-needle aspiration (FNA)specimens in patients with thyroid nodule. Methods The BRAF V600E mutation status were assessed in FNA specimens of 104 patients with thyroid nodules before operations.The BRAF mutation status,clinical,and pathology records of the patients were reviewed and the associations between these characteristics and papillary thyroid cancer (PTC ) were analyzed.Results Seventy-one PTC and 14 benign thyroid nodules were included in this study.BRAF V600E mutations were found in 57/71 (80%)PTC.All benign thyroid nodules had no BRAF V600E mutation.The sensitivity,specificity,positive predictive value and negative predictive value of BRAF V600E mutations in differentiation between PTC and benign thyroid nodules were 80%,100%,100% and 50%(P < 0.001 ).In 44 patients with PTC who underwent surgery,the central compartment lymph node metastases and extrathyroidal invasion were not significantly different between BRAF-positive and BRAF-negative PTC (P = 0.283 and 0.307 ).Conclusions BRAF V600E mutation may be a potential tool to facilitate ultrasound in diagnosis of PTC.In patients with PTC,the presence of the BRAFV600E mutation was not significantly associated with prognostic factors.

Objective To compare the clinical efficacies of vasiculation moxibustion with pastes prepared in different ways and at different acupoint groups in treating allergic rhinitis due to deficient cold of lung qi. Method Eighty subjects were randomized into 4 groups, 20 in each group. Raw Bai Jie Zi (Semen Brassicae) was used in the vesiculation paste for group 1 and 2;stir-baked Bai Jie Zi (Semen Brassicae) was used for group 3 and 4. In group 1 and group 3, bilateral Feishu (BL 13), Fengmen (BL12), and Gaohuang (BL 43) were selected; in group 2 and group 4, bilateral Dazhu (BL11), Geshu (BL17), and Shenshu (BL23) were selected. The symptom and sign scores were compared before and after intervention. Result There were significant differences in comparing the symptoms and signs among the four groups (P＜0.05), and paired comparisons showed that the therapeutic efficacies of group 1 and group 3 were significantly higher than that of group 2 and group 4 (P＜0.05), while the differences in the therapeutic efficacy between group 1 and group 3, group 2 and group 4 were statistically insignificant (P>0.05). Conclusion Vesiculation moxibustion at the point group of Fengmen (BL12), Feishu (BL13), and Gaohuang (BL43) can markedly improve the symptoms and signs of allergic rhinitis due to deficient cold of lung qi;raw and stir-baked Bai Jie Zi (Semen Brassicae) can work equally in vesiculation paste for allergic rhinitis.

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.