Objective To study the clinical value of the combination detection of serum tumor markers alpha fetal protein hetero-geneity-L3(AFP-L3) ,Golgi protein 73(GP73) and phosphatidylinositol proteoglycan-3(GPC-3) in the diagnosis of primary hepatic cancer .Methods The serum AFP-L3 ,GP73 and GPC-3 levels were measured in 34 patients with primary hepatic cancer (PHC) ,20 patients with liver cirrhosis ,20 patients with chronic hepatitis B ,37 patients with other tumors by the enzyme linked immunosor-bent assay(ELISA) .Meanwhile ,20 individuals with healthy physical examination were selected as the control group .The expression situation of various indexes were compared and analyzed .Results The expression levels of AFP-L3 ,GP73 and GPC-3 in the PHC group were significantly higher than those in the control group and the other tumors groups ,difference was statistically significant (P<0 .05) .The areas under the receiver operating characteristic (ROC) curve of PHC in the single item detection of 3 markers were 0 .909 ,0 .832 and 0 .817 respectively .The areas of the ROC curve in the combination examinations of two items were 0 .935 , 0 .945 and 0 .912 respectively .The area of the ROC curve in the 3-item combined detection of PHC could be up to 0 .960 .Conclusion The combined detection of AFP-L3 ,GP73 and GPC-3 can increase the detective rate of PHC and has important clinical significance to early diagnosis of PHC .

Objective To investigate the clinical value of serum tumor marker alpha-fetoprotein heterogeneity 3 (AFP-L3),golgin 73 (GP73) and glypican 3 (GPC-3) in primary hepatic cancer by ROC curve.Methods The AFP-L3,GP73,GPC-3 levels in serum were detected in 34 patients of primary hepatic cancer by enzyme immunoassay.Meanwhile,same terms from 20 healthy volunteers were detected as normal group.The area under curve was made by SPSS 17.0.Each laboratory indicator levels were compared and analyzed.Results The AFP-L3,GP73,GPC-3 levels in primary hepatic cancer group were significantly higher than those in normal control [(1890.13±506.47) ng/L vs (623.40±317.89) ng/L,(219.53±136.33) ng/ml vs (56.40± 25.63) ng/ml,(14.28±7.15) μg/L vs (7.33±3.71) μg/L,P ＜ 0.01].The areas under the concentration-time curve of receiver operating characteristic of single tumor marker were 0.909,0.832,0.817,the areas of AFP-L3+ GP73,AFP-L3+GPC-3,GP73+GPC-3 were 0.935,0.945,0.912,the area of combined detection of primaryhepatic cancer was 0.96.Conclusion AFP-L3 combines with GP73 and GPC-3 can increase the positive rate in patients with primary hepatic cancer and has important clinical significance.

AIM: To evaluate the safety and efficacy of ultrasonic cycloplasty(UCP)combined with anti-vascular endothelial growth factor(VEGF)+ panretinal photocoagulation(PRP)in the treatment of advanced neovascular glaucoma(NVG).METHODS: Retrospective study. A total of 45 patients(45 eyes)with advanced NVG who received surgery in our hospital from August 2020 to September 2022 were collected and divided into UCP+ anti-VEGF +PRP group(16 patients, 16 eyes), transscleral cyclophotocoagulation(TCP)+anti-VEGF+PRP group(20 patients, 20 eyes), UCP alone group(9 patients, 9 eyes). The intraocular pressure, pain scores, postoperative medication, effective rate, total success rate and the incidence of complications of the patients in the three groups were compared before surgery and at 1 d, 1 wk, 1 and 3 mo after surgery.RESULTS: There was no significant difference in preoperative intraocular pressure, pain scores and preoperative medication of patients in the three groups(all P>0.05). While there were statistical significance in the intraocular pressure and pain scores at 1 d, 1 wk, 1 and 3 mo after surgery(all P<0.01). The intraocular pressure of the UCP alone group(31.78±10.23 mmHg)was found to be higher than that of both the UCP+ anti-VEGF +PRP group(19.44±8.23 mmHg)and the TCP+ anti-VEGF +PRP group(20.80±10.27 mmHg)at 1 mo postoperatively(all P<0.017). The pain score of the TCP+ anti-VEGF +PRP group at 1 d and 1 wk postoperatively was higher than both the UCP+ anti-VEGF +PRP group and the UCP alone group(all P<0.017). The effective rates of UCP+ anti-VEGF +PRP group, TCP+ anti-VEGF +PRP group and UCP alone group were 81%(13/16), 75%(15/20)and 67%(6/9), respectively,(P=0.675), and the success rates were 69%(11/16), 50%(10/20), and 0(0/9), respectively(P=0.003). There was no significant difference in complications of patients in the three groups(P>0.05).CONCLUSION: UCP combined with anti-VEGF +PRP and TCP combined with anti-VEGF +PRP showed comparable efficacy in reducing intraocular pressure in advanced NVG. UCP combined with anti-VEGF+PRP was more effective in relieving pain and with no serious complications in advanced NVG. UCP alone can effectively control intraocular pressure and alleviate the pain of patients in the early postoperative period, but long-term control still requires anti-VEGF+PRP.

Objective:To explore the role of NKG2D ligand MHC-I related molecule A (MICA) in chemotherapy combined with NK cell immunotherapy in patients with advanced esophageal cancer after surgery. Methods:A total of 90 patients with esophageal cancer from Fujian Provincial Tumor Hospital were divided into three groups after surgery:40 patients of chemotherapy alone, 25 patients of chemotherapy combined with NK cell therapy with negative expression of MICA (MICA-group), and 25 patients of chemotherapy combined with NK cells therapy with positive expression of MICA (MICA+group). The efficacy was then compared. Results:Compared with the chemotherapy alone and MICA-groups, the positive rates of CD3+, CD4+T cells, NK cells, and the CD4+/CD8+ratio in peripheral blood from MICA+group were higher than those before treatment (64.2%± 6.4%vs. 51.3%± 5.6%, 39.8%± 8.2%vs. 29.5%± 3.2%, 25.3%± 2.1%vs. 16.4%±4.3%, 1.4%± 0.5%vs. 1.1%± 0.7%;P<0.05). Meanwhile, the levels of T-reg cells were lower than those before treatment (6.3%± 4.5%vs. 17.3%± 2.4%, P<0.05). No significant difference was observed between the disease control rate and response rate. Chemotherapy-induced neutropenia and peripheral neurotoxicity symptoms were significantly improved, and time to progression (TTP) and overall survival (OS) were significantly prolonged (P<0.05). No statistically significant difference was observed between the chemotherapy alone group and MICA-group (P>0.05). Conclusion:Treatment with chemotherapy and autologous NK cells on patients with advanced esophageal carcinoma and MICA positive expression can be safely transfused with only minor side effects and can effectively improve a patient's immune system, quality of life, and survival.

Objective To evaluate the clinical effects of cytokine-induced killer (CIK) cells combined with chemotherapy on the treatment of patients with advanced colorectal cancer.Methods CIK cells were prepared from 50 ml peripheral blood mononuclear cells by stimulated with IL-2,IFN-γ,anti-CD3 monoclone antibody,IL-1 for 8 d.The clinical effects and survival rate were compared between CIK cells combined with chemotherapy group and the chemotherapy group (50 patients with advanced colorectal cancer for each).T cells and NK cells of patients were tested by FCM before and after CIK cells treatment.The improvement of quality of life and toxicity of this therapy were observed.Results The percentages of CD3+,CD4+,CD8+ T cells and NK cells were (54.779±14.228) ％,(30.821±11.554) ％,(16.676±6.256) ％,(18.705±9.347) ％ before CIK cells transfusion.After transfusion,the percentages were (65.236±14.901) ％,(37.292±8.880) ％,(25.229±6.711) ％,(22.950±8.933) ％,respectively.The percentages were expanded greatly (P ＜ 0.05).The patients quality of life were improved clearly with lower toxicity.The DCR of CIK cells combined with chemotherapy group (64 ％,32/50) was higher than the chemotherapy group (40 ％,20/50) (P ＜ 0.05).The survival rate between two groups had no statistical significance (P ＞ 0.05).Conclusion Administration of CIK cells combined with chemotherapy can enhance immune function in patients with advanced colorectal cancer and improve their quality of life,and get good clinical efficacy.

Objective:To investigate the cytotoxic effects of CTL cell induced by DCs loaded with exosomes derived from hepatoma Huh-7 cells(T-exo).Methods: Exosomes derived from hepatoma Huh-7 cells were isolated and purified by combination of ultrafiltration centrifugation and sucrose density gradient centrifugation.Morphology of exosomes was observed under transmission electron microscopy and the expression of CD 9,CD63,HSP70 and AFP was detected by Western blot.DCs were induced with peripheral blood monocytes isolated from healthy donors.Flow cytometry was used to analysis surface markers of the DCs loaded with T-exo.WST-1 light absorption measurement was adopted to evaluate the T cell proliferation ability.Annexin-V/PI Flow cytometry were respectively used to examined cytotoxicity against the tumor cells.Results:Exosomes isolated and extracted from culture supernatant of Huh-7 cells presented as circular or elliptical vesicle with bilayer membrane , unequal in size , and with diameter of 50 to 100 nm.Western blot showed that the T-exo expressed CD9,CD63,HSP70 and AFP molecules.DCs loaded with T-exo caused significantly higher T cell pro-liferation and cytotoxic effect against AFP positive Huh-7 cells as compare to gainst AFP negative SMMC 7721 cells and un-loaded control group ( P<0.05 ).Conclusion: T-exosome loaded-DC can promote proliferation and induce significant cytotoxic effect of CTL against Huh-7 cells.

Background and purpose: Renal-cell carcinoma (RCC) is susceptible to immune therapy including the use of the nonmyeloablative allogeneic transplantation(NAT). However, NST can produce severe toxicity, so it might not be appropriate for many patients with metastatic RCC. Other novel allogeneic immunotherapies have been designed to induce an autologous immune response directed against the malignancy. This study evaluated the efficacy and safety of infusions of partially HLA-matched irradiated allogeneic blood mononuclear cells for advanced renal-cell carcinoma. Methods: Patients with histologically proven diagnosis of advanced RCC received infusions of partially HLA-matched allogeneic blood mononuclear cells. Repeat infusions were given every 8 weeks. Treatment was continued until disease progressed, unacceptable toxicity, or patient (or donor) choice. Results: Eight patients were enrolled. After every infusion, 6 patients received an oral administration of thalidomide daily with 100-300 mg/d for 2 months. One patient had durable complete response. Five stable diseases and two progress diseases were observed. In eight patients, time to progression and survival were 320 and 879+days, respectively. Severe toxicity was not observed. Conclusion: Infusions of partially HLA-matcbed irradiated allogeneic blood mononuclear cells for advanced RCC may induce some antitumor effects and deserves further study.

[摘 要] 目的：制备并表征包载CPI-444并偶联CD8抗体的纳米微粒（CNP/αCD8），探讨其对CD8+ T细胞活化、增殖和抗肿瘤作用的影响。方法: 采用复乳溶剂蒸发法和EDC/NHS法制备包载腺苷受体A2A（A2AR）特异性拮抗剂CPI-444（C）或香豆素6（C6）荧光素的纳米微粒并分别在其表面偶联CD8抗体，制得CNP/αCD8和C6NP/αCD8。扫描电镜和NanoPlus粒度测定仪表征纳米微粒形态和粒径，液相色谱与串联质谱联用（LC-MS/MS）法和离心法测定纳米微粒的载药量和药物释放情况，荧光显微镜和流式细胞仪检测CD8+ T细胞内化C6NP/αCD8的情况，流式细胞仪、ELISA和LDH法检测CNP/αCD8对CD8+ T细胞增殖、活化、细胞毒活性和杀瘤能力的影响。结果: CNP/αCD8纳米微粒为圆形、粒径约150 nm，能有效包载CPI-444和偶联CD8抗体，药物包封率和CD8抗体偶联效率分别约为60%和53.4%；CNP/αCD8纳米微粒具有良好稳定性，能被CD8+ T细胞内化，抑制A2AR分子表达。生物学功能实验显示，CNP/αCD8增强CD8+ T细胞的增殖能力、促进T细胞活化、分泌细胞因子及产生颗粒酶B和穿孔素，并增强CD8+ T细胞杀伤肿瘤细胞的能力。结论: CNP/αCD8纳米微粒能显著增强CD8+ T细胞免疫效应功能，其增强CD8+ T细胞功能可能是通过抑制A2AR分子的表达起作用。

Objective: : To investigate the effects of GBX2 gene on the proliferation, migration and invasion of human cervical carcinoma SiHa cells and to explore the mechanism. Methods: Recombinant plasmid over-expressing GBX2 gene (pCMV6-entry-GBX2, experimental group) and empty vector plasmid (pCMV6-entry, negative control group) were transfected into cervical cancer SiHa cells by plasmid transfection technique. The proliferation, colony formation and cell cycle of transfected cells were detected by WST-1 method, Colony formation assay and flow cytometry, respectively. The cell migration and invasion were detected by wound healing assay and Transwell assay. The expression level of IL-6 in cell culture supernatant was detected by ELISA. WB was used to detect the expression changes of EMT-related proteins and to explore its possible mechanism. Results: Compared with the SiHa/pCMV6 negative control group, after up-regulation of GBX2, (1) the proliferation, colony formation, migration and invasion of SiHa/GBX2 cells in the experimental group were significantly enhanced (all P<0.01); The proportion of cells in G0/G1 phase decreased while the proportion of cells in S phase and G2/M phase increased (all P<0.01); (2) the expression of E-cadherin decreased, and the expressions of N-cadherin, vimentin and snail increased (all P<0.01); (3) the expression of IL-6 in the culture supernatant of SiHa/GBX2 cells was significantly up-regulated (P<0.01); (4) STAT3 phosphorylation in SiHa/GBX2 cells was enhanced, and could be inhibited by STAT3 inhibitor S31-201 (P<0.01). Conclusion: GBX2 may induce EMT of cervical cancer SiHa cells through IL-6/STAT3 pathway, thereby promoting the proliferation, migration and invasion of cervical cancer cells.