Changes of CD4＋ CD25＋ Regulatory T Cells in Peripheral Blood in Patients with Hepatocellular Carcinoma Before and after TACE

This study investigated the changes of CD4 CD25 regulatory T cells (Tregs) in periph-eral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoem-bolization (TACE). The proportion of CD4 CD25 Tregs among CD4 T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4 CD25 Tregs among CD4 T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4 CD25 Tregs among CD4 T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4 CD25 Tregs in peripheral blood. TACE did not significantly affect the level of CD4 CD25 Tregs within short time (such as 1 week). The proportion of CD4 CD25 Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.     

Changes in immunological function after treatment with transarterial chemoembolization plus radiofrequency ablation in hepatocellular carcinoma patients

Background Different strategies for hepatocellular carcinoma （HCC） may have distinct effects on the immune system.The aim of this research was to investigate changes in the immunological function after transcatheter arterial chemoembolization （TACE） plus radiofrequency ablation （RFA） in HCC patients.Methods A total of 51 consecutive HCC treatment-naive patients was enrolled in this study and 20 healthy subjects served as controls.The therapeutic strategy was selected according to the tumor stage and general conditions.TACE was performed in 25 cases,TACE plus RFA in 17 and RFA in nine.All the patients underwent routine examinations and peripheral blood was harvested for the detection of lymphocyte subset by flow cytometry 1 day before,and 2 and 4 weeks after the treatment.The serum levels of alpha-fetoprotein （AFP）,ALT and AST were also measured before and 4 weeks after treatment for the evaluation of therapeutic efficacy and liver function impairment.Results When compared with healthy controls,the CD4/CD8 ratio and the number of B cells and natural killer （NK） cells were significantly decreased in HCC patients before treatment （P 〈0.05）.When compared with before treatment,the CD4＋ cells and CD4/CD8 ratio decreased but CD8＋ cells increased in the TACE group （P 〈0.05）; the CD4/CD8 ratio and NK cells decreased but CD8＋ cells increased in the TACE-RFA group （P 〈0.05）; the CD3＋ cells,CD4＋ cells,CD4/CD8 ratio and NK cells increased in the RFA group （P 〈0.05）.Significant differences in the CD3＋ cells,CD8＋ cells,CD4/CD8 ratio and NK cells were observed among groups （P 〈0.05）.Moreover,the AFP level decreased and transaminase level increased in all groups （P 〈0.05）.Differences of pre and post treatment between groups were statistically significant （P =0.016,0.025,0.018 respectively）.Conclusions Immunity was compromised in HCC patients; TACE and TACE plus RFA lowered immunologic function to a certain extent.RFA improved it accompanied by a protective effect on liver function.     

Changes of CD4^＋CD25^＋ Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

The function of CD4＋CD25＋ regulatory T lymphocytes （Treg） in patients with acute coronary syndrome （ACS） and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups： group C receiving conventional therapy （n=24）, and group C＋A receiving conventional therapy＋atorvastatin （10 mg/day, n=24）. T lymphocytes from ACS patients （before and 2 weeks after the treatment） or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction （MLR）. ELISA was used to measure the serum levels of cytokines （IL-10, TGF-β1 and IFN-γ） before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly （P〈0.01）, the inhibitory ability of Treg on the T lymphocytes proliferation was reduced （P〈0.01）, IFN-γ levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvastatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were significantly increased in ACS patients. Serum IFN-γ was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory effects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restoration of immune homeostasis.     

CD4^＋CD25^high Regulatory Cells in Peripheral Blood of NSCLC Patients

The proportion and changes of CD4 CD25high regulatory T cells (Trs) in peripheral blood of non-small cell lung cancer (NSCLC) patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets (CD3 , CD4 and CD8 ) and CD4 CD25high Tr cells. The results showed that the proportion of CD4 CD25high Tr cells in NSCLC group was significantly higher than in control group [(4.36±2.07) % vs (2.04±1.03) %, P<0.01]. The proportion of CD4 CD25 high Tr cells in late stage was higher than that in early stage [stages I II (2.26 0.6) %; stage III (3.28 1.38) %; stage IV (6.06±4.08) %] (P<0.05). Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4 CD25high Tr cells in peripheral blood was worse (P=0.0026). In conclusion, the relative increase in CD4 CD25high Tr cells in peripheral blood may be related to im-munosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4 CD25 high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4 CD25high Tr cell therapy to treat NSCLC patients may be an effective strategy.     

Effect of Tiotropium Bromide on Expression of CD_8~＋CD_（25）~＋FoxP_3~＋ Regulatory T Cells in Patients with Stable Chronic Obstructive Pulmonary Disease

The expression of CD8+CD25+FoxP3+ regulatory T cells(CD8+Tregs) in the peripheral blood of patients with stable chronic obstructive pulmonary disease(COPD),and the effect of muscarinic cholinergic receptor antagonist tiotropium bromide on the expression of CD8+Tregs were investigated.Twenty-three patients with moderate to severe stable COPD were enrolled in this study.All patients inhaled tiotropium bromide(18 μg daily) for 3 months.Before and after inhalation of tiotropium bromide,peripheral blood samples were collected from the patients,and T cells were labeled by three-color labeled monoclonal antibodies.Flow cytometry was used to detect the quantity and percentage of CD8+T cells,CD8+CD25+T cells,CD8+Tregs,CD4+T cells,CD4+CD25+T cells and CD4+CD25+FoxP3+ regulatory T cells(CD4+Tregs) respectively.The percentage of CD4+T cells was increased from(27.82±2.18)% to(35.53±1.3)%(t=3.20,P=0.004) in the peripheral blood of patients with stable COPD after inhalation of tiotropium bromide for 3 months,that of CD4+CD25+T cells was decreased from(10.03 ±1.42)% to(4.21 ±0.65)%(t=3.78,P=0.001),and that of CD8+Tregs was increased from(8.41 ±1.68)% to(21.34 ±4.20)%(t=2.72,P=0.013).At baseline,CD8+T cells,CD8+CD25+T cells and CD4+Tregs were detectable in the peripheral blood,but no significant changes were observed after treatment.Linear correlation analysis revealed that the difference before and after treatment in CD4+T cells and CD4+CD25+T cells was negatively correlated with the ratio of change in CD8+Tregs before and after treatment(r=-0.61,P=0.013;r=-0.72,P=0.001 respectively).In the peripheral blood of patients with stable COPD,there was the expression of CD8+Tregs and CD4+Tregs.Muscarinic receptor antagonist,tiotropium bromide,can promote the amplification of CD4+T cells,inhibit the expression of CD25+T cells,and enhance the expression of CD8+Tregs.CD8+Tregs and CD4+Tregs can be used as new indicators to understand the immune status of patients.They are helpful in judging the treatment efficacy and disease immunophenotype.     

Changes of Regulatory T Cells in Graves＇ Disease

The immune mechanism of Graves＇ diseases （GD） and the roles of regulator T cells were investigated. In 32 patients with GD （GD group） and 20 healthy volunteers （control group）, flow cytometry was used to detect the proportion of CD4^＋CD25^＋ cells, MACS to isolate CD4^＋ CD25^＋ cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the leyel of IL-10, respectively. It was found that there was no significant change in the proportion of CD4^＋CD25^＋ T cells between GD group and control group （P〉0.05）, while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group （P〈0.01 and P〈0.05, respectively）. In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.     

Relationship between the Changes of VEGF Level and Dendritic Cells in Peripheral Blood of Patients with Hepatocellular Carcinoma after Transcatheter Arterial Chemoembolization

In order to investigate the relationship between the VEGF level and the counts of den-dritic cells (DCs) in peripheral blood of patients with hepatocellular carcinoma (HCC) before and af-ter transcatheter arterial chemoembolization (TACE), the peripheral blood was obtained from 37 pa-tients with HCC who treated by TACE. The blood was obtained on the day before TACE, the first day, the 7th day and the 15th day after TACE respectively. The counts of DCs were quantified by flow cytometry. The plasma VEGF level was measured by ELESA kit. It was shown after TACE, the counts of DCs in peripheral blood were decreased significantly (P<0.05), and the VEGF level in pe-ripheral blood was increased significantly (P<0.05). The counts of DCs in peripheral blood had an inverse correlation with the plasma VEGF level (r=-0.57, P<0.05) after TACE. It was concluded that in patients with HCC after TACE, the increased plasma VEGF level appeared to have the effect to suppress the maturation of DCs, which may contribute to reduction of the body's anti-tumor im-munity effect, with a consequence of recur and metastasis of tumor.     

Analysis of CD4^＋CD25^＋ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4 CD25 regulatory T cells (CD4 CD25 Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4 CD25 Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs of exac-erbation and persistent groups were lower than that of remission and normal control groups (P<0.05). Although the CD4 CD25 Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them (P>0.05). As compared with persistent group, exacerbation group had lower CD4 CD25 Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4 CD25 Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.     

Changes of Regulatory T Cells in Graves' Disease

The immune mechanism of Graves' diseases (GD) and the roles of regulator T cells were investigated. In 32 patients with GD (GD group) and 20 healthy volunteers (control group), flow cy-tometry was used to detect the proportion of CD4 CD25 cells, MACS to isolate CD4 CD25 cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the level of IL-10, respectively. It was found that there was no significant change in the proportion of CD4 CD25 T cells between GD group and control group (P>0.05), while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group (P<0.01 and P<0.05, respectively). In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.     

Correlation of Thl7 cells and CD4＋CD25＋ regulatory T cells with clinical parameters in patients with systemic sclerosis

Background Systemic sclerosis （SSc） is an autoimmune disease that has three major components： inflammation, fibrosis, and vasculopathy. T-helper 17 cell （Th17） and regulatory T cell （Treg） are considered to be critical for autoimmune disease pathogenesis. The role of Th17 and Treg in SSc is still unclear. The aim of this study was to detect the presence of Th17s and CD4＊CD25~ Tregs in peripheral blood samples from SSc patients and to investigate the possible roles of these two T cell subsets in SSc pathogenesis. Methods Th17s （CD4 and IL-17 positive） and CD4＊CD25~ Tregs （CD4, CD25 and Foxp3 positive） in the peripheral blood mononuclear cells of 53 SSc patients and 27 healthy controls were counted by flow cytometry. The differences between SSc and control patients were analyzed. Clinical parameters, including disease duration, duration of the second symptoms, Modified Rodnan Skin Score （MRSS）, anti-topoisomerase I antibody, anti-U1 ribonucleoprotein （RNP） antibody, systemic involvements, pulmonary function test （PFT） and high resolution computed tomography （HRCT） score were prospectively collected following EUSTAR （EULAR scleroderma trial and research group） protocols. The correlations between the experimental and clinical data were investigated. Results The ratio of Th17 in SSc patients was significantly elevated compared to healthy controls （8.74% vs. 4.41%, P 〈0.001）. The amount of Th17 was positively correlated with disease duration （R=-0.531, P=-0.013） and duration of the second symptoms （R=-0.505, P=0.023）. The ratio of CD4＊CD25＊ Treg in SSc patients also significantly differed from the healthy controls （3.04% vs. 2.24%, P=0.018）. Elevated Tregs were more frequently observed in patients with a high interstitial lung disease （ILD） score on computed tomography （24/36） compared with patients with normal ILD scores （4/12, ,P=-0.043）. Elevated Tregs were also more often observed in patients with low carbon monoxide diffusing capacity     

Foxp3 expression in CD4<Superscript>+</Superscript>CD25<Superscript>+</Superscript>Foxp3<Superscript>+</Superscript> regulatory T cells promotes development of colorectal cancer by inhibiting tumor immunity

The mechanism underlying CD4⁺CD25⁺Foxp3⁺ regulatory T cells (Tregs) promoting the development of colorectal cancer (CRC) was elucidated in the present study. Forty-eight cases of colorectal carcinomas, 22 cases of colon polyps and 21 cases of normal colorectal tissues were collected. The correlation among Foxp3, IL-10 and Stat3, and the clinical relevance of these three indexes were analyzed. The results showed that the levels of Foxp3 expressed in infiltrating CD4⁺CD25⁺Foxp3⁺Tregs, and IL-10 and Stat3 in CRC tissues were all significantly higher than those in polypus tissues and normal colon tissues (P< 0.01). Pearson correlation analysis indicated that the expression level of Foxp3 was positively correlated with Stat3 at mRNA level (r=0.526, P=0.036), and was positively correlated with IL-10 at protein level (r=0.314, P=0.030). The Foxp3 expressed in CD4⁺CD25⁺Foxp3⁺Tregs was correlated with the histological grade, lymph node metastasis and TNM stage of CRC (P<0.05 for all). The IL-10 expression was correlated with the histological grade and TNM stage (both P<0.05). The Stat3 expression was correlated with the lymph node metastasis and TNM stage (both P<0.05). It was concluded that CD4⁺CD25⁺Foxp3⁺Tregs can inhibit tumor immunity in combination with some other related inhibitory cytokines and that Foxp3 expression in CD4⁺CD25⁺Foxp3⁺Tregs correlates with CRC progression.     

Cyberknife立体定向放射治疗对恶性肿瘤患者外周血MDSCs和Tregs影响

目的 动态监测恶性肿瘤患者Cyberknife大剂量分割放射治疗后外周血髓系抑制细胞(MDSCs)和调节性T细胞(Tregs)的变化。 方法 收集25例(152个样本)Cyberknife立体定向放射治疗恶性肿瘤患者放疗前(Before RT)、放疗结束(After RT)、放疗后1周(After 1W)和放疗后1月(After 1M)外周血标本,并在CT/MR图像上测量Before RT组和After 1M组肿瘤最大直径。流式细胞术检测 CD11b⁺CD33⁺HLA-DR^-/low MDSCs在单核细胞中所占比率和CD4⁺CD25⁺CD127^-/low Tregs占外周血CD4⁺细胞的百分比; 分析MDSCs和Tregs在不同时间点的变化和两者之间的相关性; 以实体瘤疗效评价标准评估疗效。 结果 MDSCs和Tregs所占比率(%):Before RT组(4.12±1.22)%和(8.57±3.72)%,After RT组(4.05±1.39)%和(8.27±3.68)%,After 1W组(3.82±0.79)%和(7.97±3.67)%,After 1M组(4.60±1.37)%和(11.16±3.67)%。外周血中的MDSCs和Tregs在Cyberknife大分割放疗后减少,1周后最低,1月后急剧增加,高于放疗前。组间配对样本t检验:After 1M组分别与Before RT组(MDSCs P=0.046,Tregs P=0.005)和After 1W组(MDSCs P=0.011,Tregs P=0.024)比较,差别有统计学意义。Before RT与After RT组之间变化MDSCs和Tregs呈正相关(P=0.032)。实体瘤疗效评价标准:稳定患者12例(48%),部分缓解患者10例(40%),完全缓解患者1例(4%),进展患者2例(8%)。Spearman相关性分析:Before RT组与After 1M组的肿瘤最大直径变化与After 1M组的MSDCs水平呈负相关(P=0.040)。 结论 Cyberknife可作为大剂量分割模型治疗恶性肿瘤; 立体定向放射治疗影响恶性肿瘤患者外周血中MDSCs和Tregs的变化,动态监测MDSCs和Tregs可能在评价恶性肿瘤患者机体免疫抑制状态和最佳免疫干预时间点选择方面具有潜在的临床应用价值。     

Liposome-mediated functional expression of multiple drug resistance gene in human bone marrow CD34<Superscript>+</Superscript> cells

Summary The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34⁺ cells was observed. Human normal bone marrow CD34⁺ cells were enriched with magnetic cell sorting (MACS) system, and then liposome-mediated MDR1 gene was transferred into bone marrow CD34⁺ cells. Fluorescence-activated cell sorter was used to evaluate the expression and functional activity of P-glycoprotein (P-gp) encoded by MDR1 gene. It was found that the purity of bone marrow CD34⁺ cells was approximately (91±4.56) % and recovery rate was (72.3±2.36) % by MACS. The expression of P-gp in the transfected CD34⁺ cells was obviously higher than that in non-transfected CD34⁺ cells. The amount of P-gp in non-transfected CD34⁺ cells was (11.2±2.2) %, but increased to (23.6±2.34) % 48 h after gene transfection (P<0.01). The amount of P-gp was gradually decreased to the basic level one week later. The accumulation and extrusion assays showed that the overexpression of P-gp could efflux Rh-123 out of cells and there was low fluorescence within the transfected cells. The functional activity of P-gp could be inhibited by 10 μg/ml verapamil. It was suggested that the transient and highly effective expression and functional activity of P-gp could be obtained by liposome-mediated MRD1 transferring into human normal bone marrow CD34⁺ cells. CAO Wenjing, female, born in 1968, Doctor in Charge     

肝动脉化疗栓塞术对原发性肝癌合并HIV/AIDS患者T淋巴细胞亚群的影响

目的 探讨原发性肝癌(primary hepatic carcinoma,PHC)合并人类免疫缺陷病毒(human immunodeficiency virus,HIV)/获得性免疫缺陷综合征(acquired immune deficiency syndrome,AIDS)患者肝动脉化疗栓塞术(transcatheter arterial chemoembolization,TACE)前后T细胞亚群的变化.方法 采用回顾性病例对照研究,选取2014年7月至2016年6月在成都市公共卫生临床医疗中心就诊的17例PHC合并HIV/AIDS患者为试验组,同期HIV阴性的PHC患者32例为对照组.采用流式细胞术(FCM)检测两组患者术前1 d及试验组患者术后1、4周外周血CD3、CD4、CD8 T细胞及CD4/CD8的比值,比较两组患者术前1 d及试验组术后1、4周外周血T细胞亚群各指标的动态变化.结果 试验组术前外周血T细胞亚群CD3、CD4、CD4/CD8比值明显低于对照组,而CD8明显高于对照组(P<0.05);试验组治疗1周后,外周血T细胞亚群CD3、CD4、CD4/CD8比值明显较治疗前下降,CD8明显增高(P<0.05),而4周后外周血T细胞亚群CD3、CD4及CD4/CD8比值较治疗1周后明显增高,CD8明显下降(P<0.05).结论 TACE对PHC合并HIV/AIDS患者是安全、有效的治疗方法.PHC合并HIV/AIDS患者细胞免疫较HIV阴性的PHC患者明显低下,TACE治疗PHC合并HIV/AIDS患者,细胞免疫状态呈现先抑制后恢复的趋势.     

Toll样受体4在乙型肝炎相关性肝癌患者外周血CD14+单核细胞表面的表达及意义

目的 通过观察TLR4在乙型肝炎相关性肝癌患者外周血CD14⁺单核细胞表面的表达,探讨其在疾病发生、发展中的作用。方法 实验共选取慢性乙型肝炎患者25例（慢性乙型肝炎组）、乙型肝炎并肝癌23例（乙型肝炎相关性肝癌组）、健康对照16例（对照组）,利用流式细胞术检测研究对象外周血CD14⁺单核细胞表面TLR4阳性细胞表达百分率和平均荧光强度的几何均数（GMF）,并分析TLR4阳性细胞表达百分率和GMF与内毒素水平及CRP、PCT的相关性。结果 乙型肝炎相关性肝癌组TLR4阳性细胞表达百分率和GMF分别为79.56%±8.12%、54.14±12.28,较慢性乙型肝炎组（46.15%±5.35%、32.27±6.46）及正常对照组（14.89%±8.21%、15.21±6.87）显著升高,差异有统计学意义（P<0.05）。乙型肝炎相关性肝癌患者TLR4阳性细胞百分率和TLR4 GMF与内毒素水平呈正相关（P<0.05）,与临床指标PCT、CRP无显著相关性（均P>0.05）。结论 TLR4在乙型肝炎相关性肝癌患者外周血CD14⁺单核细胞表面表达有显著升高,提示TLR4可能在乙型肝炎相关性肝癌的发生和发展中具有促进作用。     

Apoptosis of Leukemia Cells Induced by CD34^＋ Cells Transferred Exogenous Fas Ligand

Summary To assess the value of CD34⁺ cells transferred exogenous Fas ligand (FasL) in inducing apoptosis of human leukemic cells, the CD34⁺ cells transfected with FasL or without, pretreated with mitomycin C, was mixed with leukemic cell line U937 cells in presence or absence of daunorubicin (DNR) or cytosine arabinoside (Ara-C). After 18 h, apoptosis of cells was detected by FCM and TUNEL. Induced for 18 h by CD34⁺ cells transfected with FasL or without, the ratio of apoptosis of U937 cells was (5.0±1.3)%, (10.8±0.6)% (P<0.01), respectively. Induced by FasL⁺CD34⁺+DNR, FasL⁺CD34⁺+Ara-C, the ratio was (13.4±1.0) % (P<0.05), (17.9±1.3)% (P<0.01), respectively. The result demonstrated that CD34⁺ cells transfected with exogenous FasL could induce apoptosis of human leukemic cells and showed a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs, suggesting that it was possible to develop a new method in treatment of leukemia. XIAO Juan, Female, born in 1974, Doctor in Charge This project was supported by the grant of National Nature Science Foundation of China (Serial No. 39770767).