[Abstract] T cell receptors (TCR) are specifically expressed on T cell surface, which can recognize different tumor antigens to kill and scavenge cancerous cells. TCR-engineered T cells (TCR-T) therapy is to harbor TCR specific to tumor cells and modify the T cells with genetic engineering techniques to achieve the purpose of treating tumors after transfusion. Despite some achievements in TCR-T therapy, there are still some problems, such as treatment toxicity, limited T cell infiltration and antigen-specific deficiency and so on. So, the safety and effectiveness of TCR-T therapy need to be constantly optimized. Therefore，this paper summarizes the research status of TCR-T therapy for solid tumors in domestic and overseas, as well as the existing problems and countermeasures.

[Abstract] Objective: To explore the role of adhesion molecule with Ig like domain 2 (AMIGO2) in the proliferation of nasopharyn‐ geal carcinoma (NPC) cells and its mechanisms. Methods: A total of 10 NPC tissue samples and 10 normal nasopharyngeal epithelial tissue samples collected at Fujian Cancer Hospital during September 2017 and November 2017 were used for this study; in addition, NPC cell lines (CNE-1, CNE-2, SUNE-1, 6-10B, C666-1) and human immobilized nasopharyngeal epithelial cell line NP69 were also collected. The relative expression of AMIGO2 mRNAin above mentioned tissues and cell lines was detected by qPCR. Lentivirus vectors were constructed to interfere AMIGO2 mRNA expression, and qPCR was used to verify its interference efficiency. CCK-8 method, Clonal formation and Flow cytometry were performed to evaluate the effect of AMIGO2 interference on proliferation, clone formation and apoptosis of NPC cells. The influence of AMIGO2 interference on PI3K/AKT/mTOR signaling pathway and proliferation related molecular markers in NPC cells was assessed by Western blotting. Results: The results of qPCR showed that AMIGO2 was highly expressed in NPC tissues, CNE-2, and SUNE-1 cells (all P<0.01). The interference efficiency of AMIGO2 in CNE-2 and SUNE-1 cells could reach over 50%. The interfering of AMIGO2 expression significantly inhibited the proliferation and clone formation of CNE-2 and SUNE-1 cells (all P<0.01), promoted cell apoptosis (all P<0.01), reduced the phosphorylated protein expression levels of PI3K, AKT and mTOR in SUNE-1 cells (all P<0.01), as well as down-regulated the protein expressions of survivin and PCNA (all P<0.01). Conclusion: AMIGO2 may promote the proliferation as well as inhibit apoptosis of NPC cells by activating the PI3K/AKT/mTOR signaling pathway, suggesting that AMIGO2 may be a potential target for NPC therapy.

Objective:To investigate the clinical significance and correlation of arginase 1 (Arg-1) and inducible nitric oxide synthase (iNOS) expression in hepatocellular carcinoma (HCC).Methods:The expression of Arg-1and iNOS in 146 cases of hepatocellular carcinoma tissues and corresponding adjacent tissues was detected by immunohistochemistry. The clinicopathological characteristics and the correlation between the expressions and prognosis were determined by chi square test, Spearman’s rank correlation, Kaplan-Meier survival analysis and Cox regression analysis.Results:The positive rates of Arg-1 and iNOS were 18.7% (23/123) and 37.0% (54/146), respectively, which was significantly lower than the adjacent tissues [100%(146/146) and 93.8% (137/146)] and the difference was statistically significant ( χ2 = 212.521, P < 0.01, χ2 = 104.276, P < 0.01). There was a positive correlation between the both expression ( r = 0.331, P < 0.01). Arg-1 low expression was correlated with preoperative serum alpha-fetoprotein (AFP) level, tumor size, differentiation degree, histological types and Edmondson’s grade. iNOS low expression was correlated with the differentiation degree and Edmondson’s grade ( P < 0.05). Kaplan Meier survival analysis showed that in patients with recurrence-free survival (RFs), Arg-1 (+) group > Arg-1 (-) group and Arg-1 (+) iNOS (+) group > Arg-1 (+) iNOS (-) group > Arg-1 (-) iNOS (-) group ( P < 0.05). Cox multivariate analysis showed that age, tumor size, Edmondson’s grade, vascular tumor emboli were significantly correlated with RFs ( P < 0.05). Conclusion:There is a positive correlation between Arg-1 and iNOS expressions in HCC, and both may reflect the HCC malignant degree. The reduced/absent expression of both may participate in the occurrence and development of HCC. The combined detection of Arg-1 and iNOS on HCC may have certain significance for the judgment of differentiation degree and prognosis.

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.

Objective To study the influence of breast-feeding on the common neonatal comorbidities of very low birth weight (ELBW) infants.Method A retrospective study was conducted in our hospital from July 2014 to September 2015.The ELBW infants with birth weight between 1 000 g and 1 500 g were enrolled into the study.The infants were assigned into three groups according to human milk intake during hospitalization:high-volume breast feeding group (the high-volume group) (human milk equal to or greater than 75％),small-volume breast feeding group (the small-volume group) (human milk less than 75％) and premature formula group (the formula group) (exclusive formula fed during hospitalization).The general status,weight gain,the incidences of gastrointestinal dysfunction,neonatal necrotizing enterocolitis (NEC),late onset sepsis,premature retinopathy (ROP),bronchopulmonary dysplasia (BPD) and periventricular leukomalacia (PVL) during hospitalization were compared between the three groups.Logistic regression analysis was applied.Result A total of 210 cases were included in the study.32 cases in the high-volume group,73 cases in the small-volume group and 105 in the formula group.The incidence of NEC (Bell's stage Ⅱ and Ⅲ) in the high-volume group was significantly lower than the small-volume group and the formula group (6.3％ vs.24.7％,26.7％) (P ＜ 0.05).No statistical differences existed as for the incidences of gastrointestinal dysfunction,sepsis,ROP,BPD,and PVL (P ＞0.05).Logistic regression analysis showed that the odds ratio (OR) of survival without NEC in the highvolume group were 0.183 compared with the formula group and 0.204 compared with the small-volume group;and the 95％ confidence interval were 0.041 ～0.818,0.044 ～ 0.938 respectively.No statistically significant differences existed among the three groups in the growth rate of body weight,the time needed to regain birth weight,the time needed to reach total enteral nutrition and the length of hospital stay (P ＞0.05).Conclusion High-volume breast milk intake can reduce the incidence of NEC (Bell's stage Ⅱ and Ⅲ).Breast-feeding has little adverse effects on common comorbidities of ELBW infants.Enhanced breastfeeding has similar efficacy comparing with formula feeding in the catch-up growth.

AIM:To study the effcts of caspase recruitment domain membrane-associated guanylate kinase protein 3 (CARMA3) knockdown on the growth, migration and invasion of human colonic carcinoma HCT116 cells and to analyze the mechanism.METHODS:A colonic carcinoma cell line with CARMA3 over-expression was selected.The CARMA3 gene in the HCT116 cells was knocked down by lentivirus technique.After screening by puromycin, the stably-transfected HCT116-shCARMA3 cell line was constructed.CARMA3 expression at mRNA and protein levels was detected by real-time PCR and Western blot,respectively.The cell proliferation was analyzed by WST-1 assay and RTCA S16 system.The colony formation ability was measured by colony-forming assay.The cell cycle was analyzed by flow cytometry.The cell morphological changes were observed under microscope.The abilities of migration and invasion in vitro were observed by wound healing assay and Transwell assay.The changes of related molecules were determined by Western blot to explore the mechanism.RESULTS:The expression of CARMA3 at mRNA and protein levels in the HCT116 cells was the highest in the 4 colonic carcinoma cell lines.HCT116-shCARMA3 cells with stably-silenced CARMA3 gene were successfully established.Among them, HCT116-shCARMA3-93 cells showed the greatest inhibition of CARMA3 at mRNA and protein levels.Therefore,HCT116-shCARMA3-93 cells were chosen as the cell model.Compared with control group, the morphological changes of the HCT116-shCARMA3-93 cells had epithelial-mesenchymal transition (EMT) reversion.The abilities of proliferation, colony formation, migration and invasion in the HCT116-shCARMA3-93 cells were obviously suppressed (P<0.01).G0 /G1 phase proportion was increased and S phase proportion was correspondingly decreased (P<0.05).Bcl10 and NF-κB were down-regulated, and mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1)showed no change.Cyclin D1 was decreased obviously and cyclin A declined slightly.Metastasis-related mar-kers matrix metalloproteinase (MMP)-2 and MMP-9 were reduced,MMP-7 remained unchanged, while tissue inhibitor of metalloproteinase(TIMP)-1 and TIMP-2 were up-regulated.Furthermore, EMT-associated molecule E-cadherin was increased, while N-cadherin, Snail, Slug and Twist were decreased to some extent.CONCLUSION:CARMA3 has an impact on the growth,migration and invasion of colonic carcinoma cell line, which is possibly related to NF-κB signaling pathway to change cell cycle and metastasis-related markers and to regulate EMT.

Objective:To explore the association of microsatellite polymorphism of MICA gene with susceptibility to esophageal cancer.Methods: PCR-STR microsatellite genotyped technique was used to detect the polymorphism of MICA in Exon 5 in 103 cases of esophageal cancer and 84 cases of normal controls.Constructed of eukaryotic expression vector in esophageal carcinoma with high frequency of occurrence of the MICA allele.NK cells killing effect to 293T cells after alleles MICA transfected were assayed by LDH and the effect on target was 20∶1.ELISA was used to test supernatants sMICA of 293T cell after transfected.Results: Identified five allelic genes in MICA Exon 5 with esophageal cancer.Each allele and its frequency respectively were:MICA-A4(9.71%),MICA-A5(22.3%),MICA-A5.1(40.8%),MICA-A6(15.5%),MICA-A9(11.7%).MICA-A5.1 showed significant difference comparison with the control group.After 293T cell line was transfected MICA allele,MICA-A5.1 group was less sensitive to NK cytotoxicity compared to other groups[(30.4±6.3)%,P<0.05].The secretion of soluble MICA increased(135.7±6.2)pg/ml.Conclusion: Esophageal cancer was relevent with the MICA-A5.1 polymorphism of MICA Exon 5 alleles.Its risk is higher than other alleles.

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.

Objective: To explore the significance of Th17 in hepatocellular carcinoma, expecially with HBV infection.Methods:Cytometric bead array(CBA) was employed to detect 5 cytokines(IL-2,IL-4,IL-6,IFN-γ,IL-17A)from 39 tumor and non-tumor tissues of HCC and combined clinical data for comparative statistic analysis.Results:The expression of IL-2,IL-4,IFN-γin liver cancer tissue[(4.61±0.28),(3.37±0.58),(3.08±1.08)pg/ml,respectively] was significant lower than non-cancer tissue [(5.57±0.59),(3.77±0.70),(3.69±1.20)pg/ml,respectively].Otherwise,the expression of IL-6,IL-17A in cancer tissue [(280.09±254.68), (2.66±1.66) pg/ml, respectively] was higher than non-cancer [(6.58 ±1.92), (1.49 ±0.98) pg/ml, respectively].And,whatever cancer or non-cancer tissue,the expression of IL-17A in tissue[(3.45±1.86)pg/ml] with high HBV load (>1 000 U/ml) was significant higher than tissue with low HBV load[(1.97±1.16)pg/ml].Conclusion: IL-17A was highly expressed in HCC,and IL-2,IL-4,IFN-γmay inhibit its expression,and IL-6 may promote it.Hepatitis B virus infection may promote Th17 expression,thereby reducing patient′s prognosis.

Objective To develop a simple method of determination of sorafenib in serum by reversed-phase high performance liquid chromatography (RP-HPLC) and to explore its application in sorafenib therapeutic drug monitoring (TDM).Methods Sorafenib extracted by ethyl ether-petroleum (9∶1) with internal standard of erlotinib from serum was wiped off in 60 ℃ water bath.Sorafenib was redissolved by mobile buffer and analyzed by 40 μl.Chromatographic column was Symmetry Rp18 (5 μm,4.6 mm×250 mm,waters) column in normal temperature.The mobile buffer was 28 mmol/L acetate buffer (pH 5.8)-acetonitrile (37∶63).Sorafenib and erlotinib were detected in 249 nm and 335 nm,respectively.Results The concentration range of sorafenib was 0.50-20.00 μg/ml (r =0.9999).The within-day and between-day accuracies of sorafenib were less than 4.77 ％ and 8.79 ％,respectively.The average recovery rate was 98.48 ％.Sorafenib was stable in serum or after extraction.The concentrations of sorafenib in two patients were detected.Conclusion Detection of sorafenib in serum by RP-HPLC is simple and accurate,which is available to determine sorafenib in serum.The TDM of sorafenib has clinical significance.