Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.

Objective To systematically evaluate the effect of preoperative transarterial chemoembolization (TACE) on perioperative safety of patients with resectable hepatocellular carcinoma (HCC).Methods Literatures were researched using Chinese Journal Full-text Database,Wanfang database,VIP database,PubMed,Medline from December 1,1994 to May 30,2016 with the key words including “肝细胞癌,肝切除,术前化疗栓塞,经动脉化疗栓塞,liver cancer,hepatocellular carcinoma,liver resection,hepatectomy,transcatheter arterial chemoembolization,transarterial chemoembolization,preoperative” Manual retrieval was also conducted simultaneously.The randomized controlled trials (RCTs) about TACE on perioperative safety of patients with resectable HCC were received and enrolled.Patients undergoing surgery after preoperative TACE were allocated into the case group and patients undergoing first-stage resection were allocated into the control group.Two reviewers independently screened literatures,extracted data and assessed the risk of bias.Count data were described as relative risk (RR) and 95％ confidence interval (CI).Measurement data were represented as standardized mean difference (SMD) and 95％CI.The heterogeneity of the studies was analyzed using the I2 test.Results Five RCTs were enrolled in the Meta analysis,and the total sample size was 430 cases including 212 in the case group and 218 in the control group.Results of Meta analysis showed that there was no statistically significant difference in the hemihepatic resection rate between the 2 groups (RR=0.99,95％CI:0.81～ 1.20,P＞0.05).The combined resection rate of perihepatic organs in the case group was significantly higher than that in the control group (RR=3.42,95％CI:1.91-6.12,P＜0.05).Results of subgroup analysis showed that operation time and incidence of postoperative complications of patients with an average tumor diameter ＞5 cm in the case group were respectively longer and higher than these in the control group (SMD=0.31,RR=1.65,95％CI:0.06-0.57,1.01-2.69,P＜0.05).Conclusion There is no obvious effect of preoperative TACE on resectable HCC,and it can evaluated combined resection rate of perihepatic organs,operation time and incidence of postoperative complications of patients with resectable HCC and an average tumor diameter ＞ 5 cm,and also reduce the perioperative safety.

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.

OBJECTIVETo detect the expression levels of co-inhibitory molecules, including CTLA-4, LAG-3, PD-1 and CD39, on CD4⁺ T cells in peripheral blood or tumor tissues from NSCLC patients and to investigate their potential internal relationships with the progression of NSCLC.

METHODSEighty-eight patients including 53 NSCLC, 17 disease control cases and 18 healthy controls were studied. All the peripheral blood and 13 cases of tumor and tumor-adjacent tissues from surgically treated NSCLC patients were obtained. The expression levels of co-inhibitory molecules CTLA-4, LAG-3, PD-1 and CD39 were assayed by flow cytometry (FCM).

RESULTSThe ratios of CD4⁺ CTLA-4⁺ T cells, CD4⁺ LAG-3⁺ T cells, CD4⁺ PD-1⁺ T cells and CD4⁺ CD39⁺ T cells in the peripheral blood of NSCLC patients were (2.49 ± 2.43)%, (2.47 ± 3.50)%, (12.94 ± 5.96)% and (6.78 ± 5.21)%, respectively, the ratio of CD4⁺ CTLA-4⁺ T cells was significantly higher in the peripheral blood of NSCLC patients than that in the disease controls and healthy controls (P < 0.05) . The ratio of CD4(+)PD-1⁺ T cells was also highly raised in the peripheral blood of NSCLC patients than that in the healthy controls (P < 0.05). Further stratified analysis indicated that the ratio of CD4⁺ PD-1⁺ T cells was (13.21 ± 5.96)% in NSCLC patients entering stages III and IV, also significantly increased as compared with that of (11.06 ± 3.42)% in the patients undergoing stages I and II (P < 0.05). More CD4⁺ CTLA-4⁺ T cells, CD4⁺ LAG-3⁺ T cells and CD4⁺ PD-1⁺ T cells were verified in the cancer tissues (5.07 ± 2.11)%, (7.86 ± 3.24)% and (40.20 ± 18.84)%, respectively, than those in their matched peripheral blood (3.13 ± 1.01)%, (2.65 ± 1.48)% and (15.79 ± 5.69)%, (P < 0.05 for all), and especially, CD4⁺ CTLA-4⁺ T cells and CD4⁺ PD-1⁺ T cells were also highly increased than those in matched cancer-adjacent tissues (P < 0.05 for all).

CONCLUSIONSThe increased expression levels of co-inhibitory molecules CTLA-4, LAG-3 and PD-1 on CD4⁺ T cells in peripheral blood and tumor tissues may be one of the mechanisms related to immune escape of tumor cells, acceleration of disease progression and poor prognosis in NSCLC patients.

Antigens, CD ; metabolism ; Apyrase ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; metabolism ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Disease Progression ; Flow Cytometry ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Programmed Cell Death 1 Receptor ; metabolism

Objective To analyze the clinical characteristics of patients with metastatic carcinoma of bone. Methods The clinical data of 217 patients with metastatic carcinoma of bone was retrospectively analyzed. The clinical features were summarized,including primary tumor,age distribution,first symptom,time of onset and bone metastatic site. Visual analogue scale(VAS)was used to evaluate the difference of pain in patients with bone metastases before and after zoledronic acid treatment. Results The bone metastasis occurred more frequently in men(male:female = 1.28:1). The majority of primary tumors were lung cancer(51.61％)and breast cancer (15.67％),respectively. The peak ages of the patients with metastatic carcinoma of bone were between 50 and 70 years old. Most of the patients with metastatic carcinoma of bone initially showed varying degrees of pain ,some of the patients with dysfunction,pathological fracture,or local lumps as the initial symptoms,only a tiny minority of patients were inadvertently found. Onset time was often within 3 months ,which accountings for 54.84％. Bone metastases were mostly in the spine,pelvis,femur and ribs,and most of the bone metastases were multiple. The VAS score of 108 patients who with bone metastases pain treated by zoledronic acid was 0～4(1.31 ± 1.07),with statistically significant differences compared with pretherapy(P<0.01). Conclusion In order to reduce the pain of patients,and to improve the quality of life and prolong the survival time,bone metastases should be diagnosed early and given comprehensive treatment.

Objective To investigate the effects of leptin on Treg cells and the possible mecha-nism. Methods Leptin-deficient ( ob/ob) mice and homologous wild-type mice were used in this study. The percentages of Treg cells in spleen tissues and peripheral blood samples were measured by flow cytometry ( FCM) . Differences in Treg cell functionality were compared between the two groups. Splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by magnetic beads, were respectively cultured with leptin and anti-leptin neutralization antibody to evaluate the effects of leptin on Treg cells. Quantitative real-time PCR was performed to analyze the expression of Treg cell-related cytokines at transcriptional level. The levels of IL-10 and TGF-β in the supernatants of CD4+T cell culture were measured with Luminex technolo-gy. Results Compared with the wild-type mice, the ob/ob mice showed higher percentages of Treg cells in both peripheral blood samples and spleen tissues [(11. 56 ± 0. 72)％ vs (5. 47 ± 0. 81)％, (10. 16 ± 0.93)％ vs (6.29±0. 69)％]. Treg cells isolated from the ob/ob mice had stronger immunosuppressive effects on the proliferation of effector T ( Teff) cells and the secretion of TNF-α and IFN-γ than those from the wild-type mice [TNF-α:(1. 6±0. 2)％ vs (2. 4±0. 5)％, IFN-γ:(4. 3±0. 3)％ vs (7. 2±1. 2)％]. The percentages of Treg cells were decreased from (12. 2±1. 8)％ to (7. 6±0. 9)％ upon the in vitro treat-ment of CD4+ T cells from the ob/ob mice with leptin and the immunosuppressive effects of Treg cells were also weakened. However, the percentages of Treg cells were increased from (7. 8±0. 85)％ to (13. 1± 1. 5)％ upon the in vitro treatment of CD4+T cells from the wild-type mice with anti-leptin antibody and the immunosuppressive effects of Treg cells were improved as well. Moreover, the expression of Foxp3, IL-10 and TGF-β at transcriptional level and the levels of IL-10 and TGF-β in the ob/ob group were higher than those in the wild-type group. Conclusions Leptin deficiency significantly promoted the generation of Treg cells in mice and resulted in an increased expression of Foxp3, IL-10 and TGF-βat mRNA level and elevat-ed levels of IL-10 and TGF-β. The treatment of CD4+T cells with leptin might inhibit the generation of Treg cells through down-regulating the transcription of Foxp3, IL-10 and TGF-β.

Objective: To compare the surgical accuracy of the 3D printing surgical guide and traditional occlusal splint in the treatment of skeletal facial asymmetry cases. Methods: 12 facial asymmetric patients underwent joint orthognathic and orthodontics treatments were included in this research. In the 3D printing group (n=6) the pre-surgical CT scan for the skeleton and laser scan for the dentures were performed and 3D orthognathic procedures including Le FortⅠ and BSSRO combined with genioplasty were planned. The 3D printing surgical guides were manufactured and applied during surgeries. Another 6 cases were received same procedures by traditional occlusal splint techniques as the control group. Post-surgical CT reconstruction and 3D superimposition with pre-surgical planning was carried out for outcome comparison. Results: The maximal error of maxillar was 0.65 mm and average error was less than 1 mm in 3D printing group, while the maxillary maximal error was 2.11 mm and average error was greater than 1 mm in traditional group which showed the statistical significance (P<0.05). Conclusions The usage of 3D printing surgical guide could improve the orthognathic surgical precision by controlling the jaw bone in a three-dimensional way.

Objective To analyze the setup errors by cone-beam computed tomography (CBCT) for breast cancer patients who were immobilized with neck and breast thermoplastic mask and received intensity modulated radiation therapy (IMRT), and to calculate the external margins from the clinical target volume (CTV) to the planning target volume (PTV) (MPTV) of tumors. Methods Twenty-five breast cancer patients who were immobilized with neck and breast thermoplastic mask and received IMRT in the Oncology Department of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from November 2016 to June 2017 were enrolled. The position of the patients were verified by CBCT before treatment . The linear and rotation errors of the X, Y and Z axes were analyzed by online bone registration. The systematic errors (Σ) and random errors (σ) of the patients were also calculated, and then the margins from CTV to PTV margins were calculated based on MPTV=2.5Σ+0.7σ. 25 patients'height, weight, body mass index (BMI) and the maximum diameters of CTV in the lateral, longitudinal and vertical directions were recorded, and the relation between the setup errors and the above mentioned was analyzed by using Spearman method. Results A total of 174 CBCT scans for 25 breast cancer patients were completed. The group Σ were 1.40 mm, 1.50 mm and 1.20 mm, and rotation errors were 0.9°, 0.7° and 0.8° at the X, Y and Z axes, respectively. The group σ were 2.20 mm, 3.00 mm and 1.40 mm, and rotation errors were 0.7°, 0.6° and 0.7° at the X, Y and Z axes, respectively. MPTVwas recommended as 4.90 mm, 6.00 mm and 3.90 mm at the X, Y and Z axes, respectively. There was no correlation between the height, weight, BMI of the patients and the setup errors (all P ＞ 0.05). However, there was a significant correlation between the maximum lateral, longitudinal diameters of the CTV and the setup errors (rs= 0.406, P= 0.044; rs= 0.512, P= 0.009). Conclusions The neck and breast thermoplastic mask can improve the diagnostic accuracy of radiotherapy in breast cancer patients. The data of setup errors verified by CBCT can provide meaningful references for the setting of MPTV.

The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.
Adjuvants, Immunologic ; Animals ; Antibodies, Neutralizing ; Antibody Formation ; Birds ; Bursa of Fabricius ; Cytokines ; Immunity, Cellular ; Immunity, Humoral ; Influenza A Virus, H9N2 Subtype ; Influenza in Birds ; Lung ; Mice ; Peptides ; T-Lymphocytes, Cytotoxic

Objective To evaluate the effectiveness of computer aided design (CAD) and three bit printing in the management of orthognathic surgery.Methods A total of 5 cases of patients with jaw deformity were involved in this study;jaw teeth and CT scanning laser scanning hefore surgery,virtual surgery design of 3D reconstruction and fusion data were analyzed,according to the design scheme of double jaw surgery combined with genioplasty;design and 3D printing of maxillary Le Fort Ⅰ osteotomy,genioplasty titanium alloy resin osteotomy and positioning guide,sagittal split ramus osteotomy by 3D printing and plate technology were used in this approach.The postoperative results were compared with the surgical planning by three-dimensional measurement and statistical analysis.Results When the operation guide plate was applied smoothly,the maximum error for maxilla was 1.2 mm (0.3-1.2 mm),and the maximum error for genioplasty was 1.7 mm,(0.5-1.7 mm),and the mean error was less than 1 mm.Follow-up for 12 months showed no adverse reaction.Conclusions Three dimensional printing surgical guide plate can accurately provide the osteotomy information,effectively control the jaw movement,and improve the orthognathic surgery accuracy of patients with partial jaw deformity.