Abstract:Due to the tumor heterogeneity，immunosuppressive microenvironment and other factors，CAR-T cells have not shown obvious clinical efficacy in the treatment of solid tumors. For the treatment of solid tumors，great progress has been made in the preparation of CAR-T cells in recent years. This article comments the research progress and tendency of CAR-T cells in solid tumor treatment.

Abstract:Objective：The phenotype of female mice was studied by constructing Id3-Gdf9-Cre+ conditional knockout（CKO）mice to further determine the effect of differentiation inhibitor Id3 on female mice fertility. Methods：The development of follicles was detected in the ovaries of the conditional knockout mice with Id3-Gdf9-Cre+ by HE staining and follicle counting. The oocyte maturation was detected by maturation experiments in vitro and superovulation experiments in vitro. And mice fertility was detected by mating experiment. Results：The results of qPCR and Western blot showed that we had successfully knocked out Id3 in oocytes. HE staining was performed on 3 weeks and 8 weeks mice ovaries and the follicles were counted at various levels，and the follicles developed at all levels were normal. Oocyte maturation experiments and superovulation experiments showed that oocyte maturation was normal in vitro. Mating experiment showed that CKO mice have normal fertility. Conclusion：Under normal feeding conditions，conditional knockout of Id3 does not affect normal follicles development or fertility of female mice.

Abstract:Objective：The aim of this study was to construct zinc transporter 8（ZnT8）chimeric antigen receptor（CAR）expression vector，so as to provide experimental basis for studying the role of antigen-specific regulatory T cells（Tregs）in the autoimmunity of type 1 diabetes. Methods：PLVX-EGFP-ZnT8 scfv lentivector was constructed by molecular cloning technique to produce ZnT8 scfv-CAR lentivirus；the expression of green fluorescent protein（GFP）in Tregs was detected by flow cytometry to determine the efficiency of lentivirus；the proliferation of Tregs was evaluated by cell counting；the expression of CD4，CD25，Foxp3 and Helios in proliferated cells were detected by flow cytometry. Results：The PLVX-EGFP-ZnT8 scfv lentivector was successfully constructed. The titer of concentrated ZnT8 scfv-CAR lentivirus was 2.4×108 TU/μL. The expression rate of GFP was 43.2% ± 4.1% in Tregs infected with ZnT8 scfv-CAR lentivirus. After being cultured in vitro for 14 days，the ZnT8-specific Tregs proliferated（634.3 ± 92.5）times，along with high expression of Foxp3（60.4% ± 3.5%）and Helios（64.3% ± 4.8%）. Conclusion：We obtained the PLVX-EGFP-ZnT8 scfv lentivector and synthesized ZnT8 scfv-CAR lentivirus successfully；Tregs infected with ZnT8 scfv-CAR lentivirus could maintain the CAR expression；CAR-Tregs were a group of CD4+CD25+ cells with high expression of Foxp3 and Helios.

Abstract:Objective：To investigate the effect of glucagon like peptide-1（GLP-1）on the calcification of aortic valve in mice fed with high fat and cholesterol diet，and to reveal the protective effect and mechanism of GLP-1 in the occurrence and development of calcified aortic valve disease（CAVD）. Methods：Thirty six C57BL/6 mice（8 weeks old，female）were randomly divided into four groups（n=9）. The normal diet group and the GLP-1 control group were given normal diet，and the high fat diet group and the GLP-1 intervention group were fed with high fat and high cholesterol diet + vitamin D2. The GLP-1 control group and the GLP-1 intervention group were additionally treated with GLP-1. After 12 weeks，the two-dimensional echocardiographic data of mice in each group were measured，including aortic valve area（AVA），left ventricular ejection fraction（EF），valve leaf thickness（T），left ventricular end systolic diameter（LVDs），transvalvular velocity（TV）. Then the serum calcium concentrations were measured，and von Kossa staining was used to observe the calcification of valve tissue，and immunohistochemistry to detect the expression of osteopontin（OPN）and p38 protein in the aortic valve samples. Results：① In the normal diet group，the GLP-1 control group and the GLP-1 intervention group，T，TV，LVDs and serum calcium concentration were lower than those in the high fat diet group，while AVA and left ventricular EF were higher than those in the high fat diet group（P < 0.05）. ② The expressions of OPN and phosphorylated p38（p-p38）in the aortic valve samples of the normal diet group，the GLP-1 control group，the GLP-1 intervention group were significantly lower than that of the high fat diet group（P < 0.05）. Conclusion：GLP-1 can inhibit the calcification of aortic valve by inhibiting the expression of p-p38 in mice.

Abstract:Objective：To investigate the effect of loss of aryl hydrocarbon receptor（AhR） on intestinal inflammation and its potential mechanism. Methods：Mice were assigned into 4 groups：wild type control group（WT control），wild type colitis group（WT TNBS），AhR knockout control group（AhR-/- control） and AhR knockout colitis group（AhR-/- TNBS）. Acute experimental colitis was established using 2，4，6-trinitrobenzenesulfonic acid（TNBS）. Disease activity index（DAI）score，colon length and histological score were compared among different groups. The expression of regulatory T cell（Treg） specific transcription factor FoxP3 was measured by Western blot（WB） and immunofluorescence（IF）. Proportion of Treg cells in peripheral blood was detected by flow cytometry. Results：Compared with WT mice，AhR-/- mice experienced much more severe colitis after TNBS administration，manifested by more weight loss（P < 0.01），severe diarrhea and bloody stool，higher DAI score（P < 0.001），shorter colon length（P < 0.01） and poorer histological findings（P < 0.001）. Also，expression of FoxP3 in the colon and proportion of Treg cells in peripheral blood were much lower in AhR-/- TNBS mice（P < 0.01）. Conclusion：Deletion of AhR was found to aggravate experimental colitis. This effect could be associated with the decrease of Treg cells in the colon and peripheral blood.

Abstract:Objective：To explore the relationship between nuclear pore membrane protein 121（POM121）expression and pathological characteristics of patients with colon cancer and the significance of POM121 to patients’ prognosis. Methods：Immunohistochemistry was used to detected POM121 level in colon cancer tissues and para-carcinoma tissues. Based on the clinical data of patients，the connection between POM121 expression and clinical pathological characteristics of colon cancer was analyzed，as well as the significance of POM121 to clinical prognosis. Results：The positive expression rate of POM121 in colon cancer tissues（66.03%）was higher than that in para-carcinoma tissues（47.42%）with a statistically significant difference（P < 0.001）. The expression of POM121 was significantly correlated with tumor siae（P=0.041），lymph node metastasis（P < 0.001），distant metastasis（P=0.002），the AJCC staging（P=0.003），venous invasion（P < 0.001），perineural invasion（P < 0.001），preoperative carcino-embryonic antigen（P < 0.001），preoperative carbohydrate antigen 199（P=0.004），and Ki67（P < 0.001）expression，while no correlation was found between the POM121 expression and gender，age，location，histological type，or differentiation. Patients with high POM121 expression had poor overall survival rates. Genes in colon cancer tissues with high POM121 expression were enriched in aminoacyl t-RNA biosynthesis，base excision repair，DNA replication and so on. Conclusion：POM121 is highly expressed in colon cancer and is an independent risk factor affecting the prognosis of patients. POM121 has the potential to become a new prognostic markers and molecular targets for targeted therapy of colon cancer

Abstract:Objective：To investigate the expression level of long non-coding RNA（lncRNA）GM15886 in lung tissues of bronchopulmonary dysplasia（BPD）induced by hyperoxia in neonatal mice and its mechanism in BPD. Methods：High expressed GM15886 was selected by previous lncRNA microarray. IntaRNA，Multi Experiment Matrix and Ensemble database were applied to predict GM15886 target genes. Sixty-four newly born C57BL/6J mice were randomly divided into the hyperoxic group and the air group，with 32 mice in each group. Newborn mice in the hyperoxia group were exposed to 95% oxygen，while those in the air group were exposed to air. Mice were sacrificed on day 0，day 3，day 5 and day 7，respectively，and the pathological changes of pulmonary tissues were analyzed via HE staining，the expression levels of GM15886 and homeodomain-interacting protein kinase 1（HIPK1）mRNA was detected by using QPCR，the expression of HIPK1 at different time points was detected by immunohistochemistry. Results：IntaRNA predicted the end of GM15886 sequence overlaps with the gene HIPK1. The expression of GM15886 in neonatal mice in the hyperoxia group increased gradually on day 3，day 5 and day 7，（1.91±0.28，2.12±0.38，and 2.35±0.43，respectively），and the differences were statistically significant compared with day 0（all P < 0.05）. The relative expressions of HIPK1 on day 3，day 5 and day 7 were 1.16±0.33，0.92±0.31，and 3.12±0.46，respectively. The expression level on day 7 was higher than that on day 0，day 3，day 5（all P < 0.05）. The expression of HIPK1 protein changed in the same way as mRNA. Conclusion：With the extension of hyperoxic exposure time，the expression of GM15886 in lung tissues increased gradually. GM15886 might participate in the pathogenesis of BPD by relugating HIPK1 expression.

Abstract:Objective：To explore correlation between long chain non-coding RNA（lncRNA）BANCR and epidermal growth factor receptor-tyrosine kinase inhibitor（EGFR-TKI）drug resistance of non-small cell lung cancer. Methods：Mutated cell strain PC9 with EGFR exon 19 deletion and drug resistant cell strain PC9/GR induced by EGFR-TKI gefitinib were selected for research. Real time fluorescent quantitative PCR（qRT-PCR）was applied to examine expression difference of BANCR in PC9/GR and PC9. BANCR was over expressed in PC9/GR cells and then CCK-8 method was used to examine drug susceptibility of the cells to gefitinib and clone formation assay to examine cell reproductive capacity. Meanwhile，flow cytometry was adopted to examine cell apoptosis and Western blot to check expression of epithelial-mesenchymal transition related proteins. Results：① BANCR was significantly low expressed in PC9/GR cells（P < 0.05）. ②When BANCR was over-expressed in PC9/GR cells，compared with control group，half inhibition concentration（IC50）of gefitinib was reduced（P < 0.05），cell reproductive capacity was lowered，and cell apoptosis increased after being handled with gefitinib（P < 0.05）. ③ Compared with control group，Western blot showed that expression level of E-cadherin in the PC9/GR cells over-expressed BANCR was significantly elevated and expression level of N-cadherin was remarkably lowered（P < 0.05）. Conclusion：Long chain non-coding RNA BANCR can induce cell apoptosis and inhibit cell reproduction. Over expressed BANCR can increase sensitivity of PC9/GR cells to gefitinib and thus reverse drug resistance. This mechanism might work through regulating EMT process.

Abstract:Objective：To study the effects of bisphenol S（BPS）on lipid metabolism in zebrafish with high-fat diet and its potential mechanism. Methods：The effects of BPS in different concentration on lipid deposition in the blood vessels of zebrafish with high-fat diet were studied，and the changes of lipid metabolism-related proteins and genes were detected by ELISA and quantitative RT-PCR. Results：The cholesterol accumulation in the larvae blood vessels was significantly decreased under the exposure of 1 and 10 μg/L BPS（P < 0.01）. When fed with high-fat diet 42 mg/dish，the lipid accumulation in the larvae blood vessels of the BPS exposed group was also significantly decreased（P < 0.01）. The expression level of RXRα protein was significantly reduced in the 100 μg/L BPS exposed group（P < 0.05）. The gene expression of hmgcrα was significantly induced by 10 μg/L BPS exposure（P < 0.05），and the mRNA levels of ldlrα and rxrα were all significantly decreased in the 100 μg/L and 1 000 μg/L BPS exposed group（all P < 0.05）. While the expression of rorα gene was reduced after exposure of 0.1，1.0，100.0 μg/L BPS（all P < 0.05），and it was lower after exposure of 1 000 μg/L BPS（P < 0.01）. Conclusion：The effect of BPS on lipid metabolism of zebrafish larvae was related to the feeding amount of high-fat diet and abnormal expression of lipid metabolism related genes，and its mechanism may be related to rxr/ror signaling pathway.

Abstract:Objective： To investigate the correlation between total cholesterol to high-density lipoprotein cholesterol ratio（TC/HDL-C）and hepatic steatosis severity in obese patients with nonalcoholic fatty liver disease. Methods：From 2018 to 2019，a total of 98 obese patients hospitalized in our hospital were recruited. Based on the severity of hepatic steatosis diagnosed with pathological grade，patients were divided into groups with mild-grade（group A），moderate-grade（group B），and severe-grade（group C）hepatic steatosis. The correlation between general clinical characteristics and pathological grade of hepatic steatosis were analyzed by Spearman correlation analysis and logistic regression. Results：①Compared to group A，TC/HDL-C，body mass index（BMI），alanine aminotransferase（ALT），aspartate aminotransferase（AST）levels of group B and group C were significantly higher. There was no significant difference in the above indicators between group B and group C. ② Using Spearman correlation analysis，we found that TC/HDL-C was positively correlated with severity of hepatic steatosis，TC，triglyceride and other related factors，while negatively correlated with HDL-C in group A and group B（P < 0.05）. ③ Logistic regression analysis showed that there was a significant correlation between TC/HDL-C and pathological grade of hepatic steatosis in group A and group B（P < 0.05）. Conclusion：TC/HDL-C was significantly higher in patients with moderate steatosis than with mild steatosis，and was significantly correlated with the degree of hepatic steatosis.

Abstract:Objective： To investigate the dynamic change of absolute natural killer cell counts（ANKC） and its prognostic value for the treatment response in the newly diagnosed patients with chronic myeloid leukemia（CML） before and after 3 months and 12 months of tyrosine kinase inhibitors（TKI） treatment. Methods：All 80 patients with chronic-phase CML（CML-CP） diagnosed from 2014 to 2019 were treated with first-generation TKI（imatinib） or second-generation TKI（nilotinib, dasatinib）. According to the treatment response，they were divided into two groups：the optimal response group and the warning/treatment failure group，aiming to analyze the influence of ANKC and related clinical data on prognosis. The receiver operating characteristic（ROC）curve was used to optimize the cut-off values of clinical indicator. Results：After 3 months and 12 months of treatment，the efficacy of patients with ANKC≥0.14×109 /L at the first diagnosis was better than those with lower ANKC（P < 0.05）. Following 3 months of TKI treatment，ANKC in patients with optimal response was significantly lower than that at initial diagnosis（P < 0.001），whereas ANKC in patients without optimal response was significantly higher（P=0.017）. Dynamic analysis was performed on the correlation between the changes in ANKC and the efficacy in 13 patients receiving dasatinib，and we found that in the patients with optimal response，ANKC after 3 months of treatment was significantly lower than that at the initial diagnosis（P=0.025），whereas ANKC after 12 months of treatment was significantly higher than that at the initial diagnosis（P=0.015）. Multivariate analysis showed that ANKC，white blood cells count and second-generation TKI were independent factors for treatment response（P < 0.05）. Conclusion：ANKC in CML-CP patients is an effective prognostic factor for TKI treatment response，meanwhile patients with ANKC≥0.14×109/L at diagnosis can achieve an optimal response.

Abstract:Objective：This article aims to investigate the efficiency of a model combined serum lactate dehydrogenase（LDH）level and model for end-stage liver disease（MELD）MELD-LDH on evaluating the prognosis of patients with acute-on-chronic liver failure（ACLF），and to study the correlation between serum LDH and different levels of multiple system damage in order to improve the awareness and treatment of ACLF. Methods：This study used a retrospective method to collect clinical data of 119 patients with ACLF admitted to the First Affiliated Hospital of Nanjing Medical University from October 2015 to August 2019. According to the prognosis of 28 days after admission，they were divided into the survival group（n=90）and the death group（n=29）. Univariate and multivariate regression analysis methods were used to analyze the relevant factors affecting the prognosis of patients with ACLF. MELD-LDH model was established to verify its efficacy and explore the correlation between serum LDH and the levels of multiple organ damage. Results：The results showed that serum LDH levels were independent risk factors affecting the prognosis of patients with ACLF（OR=1.015，P=0.002）. LDH was significantly higher in patients with renal failure and hepatic encephalopathy（P=0.002，P=0.018）. MELD-LDH model was significantly better than MELD scores in predicting 28-day and 90-day clinical outcomes（P=0.028，P=0.002）. Conclusion：High serum LDH level is an independent risk factor for 28-day mortality in patients with ACLF，and it is closely related to renal failure and hepatic encephalopathy. MELD-LDH model established by combining LDH and MELD scores has a better predictive effect on the 28-day and 90-day prognosis of patients with ACLF.

Abstract:Objective：To investigate the perioperative effect of aortic root enlargement for adult patients with small aortic root undergoing aortic valve replacement. Methods：Twenty cases with small aortic root underwent aortic root enlargement and aortic valve replacement as the research group from 2018 to 2019，150 cases with normal aortic root underwent aortic valve replacement as the normal aortic root group，and 143 patients with small aortic root underwent aortic replacement rather than root enlargement as the small aortic root group from 2013 to 2018. The inverse probability of treatment weighting（IPTW）method was used to balance the difference of baseline parameters among the three groups，and logistic multivariate regression was used to detect the independent risk factors of postoperative increase in aortic valve pressure gradient. Results：After the IPTW，one month after aortic valve replacement，the peak aortic valve pressure gradients of the normal aortic root group，the small aortic root group，and the research group were（26.79 ± 10.85）mmHg，（36.88 ± 19.27）mmHg，and（20.85 ± 6.25）mmHg. The differences of the pairwise comparison were still statistically significant（P < 0.01）. Multivariate logistic regression analysis showed the treatment strategy of aortic root［small aortic root without aortic root enlargement，β（95%CI）：8.48（2.17~14.80），P=0.010；small aortic root underwent aortic root enlargement，β（95%CI）：-12.16（-21.79~2.52），P=0.015］and preoperative peak aortic valve pressure gradient［β（95%CI）：0.17（0.03~0.30），P=0.020］were independent risk factors for increased peak aortic valve pressure gradient. Conclusion：For patients with small aortic root，the short-term hemodynamic effect after aortic root enlargement is better，but the long-term clinical results need further follow-up.

Abstract:Objective：To investigate the short and medium-term effects of simultaneous maze Ⅳ on comorbid atrial fibrillation（AF）in patients underwent atrial septal defect（ASD）repair. Methods：The data of 28 patients with concomitant ASD and AF from February 2014 to May 2019 were analyzed retrospectively. All patients underwent ASD repair and maze Ⅳ procedure at the same time in our hospital. The surgical complications and the success rate of AF ablation from AF in medium-term follow-up was recorded. Results：The median follow-up was 2 years，with no deaths during the follow-up period. The success rates of AF ablation at 6 months，1 year，and 2 years after the operation were 85.7%，78.5%，and 78.5%，respectively. No pacemaker implantation needed due to atrioventricular block，and no stroke occurred during the follow-up period. Conclusion：ASD patients combined with AF should be treated with ASD repair and maze Ⅳ procedure at the same time，which does not increase the risks of surgery. The rates for maintenance of sinus rhythm in short-term and medium-term are satisfactory although the long-term results needs further research.

Abstract:Objective：To investigate the effect of 1% cyclopentolate on the power and axis of astigmatism in 5~6 years old preschool children. Methods：From September 2017 to March 2018，a cross-sectional study was carried out in 1 920 children born between September 2011 and August 2012 in Yuhuatai District. The noncycloplegic refractive status inculding spherical power，astigmatic power，and axis angle were measured using an autorefractor Canon RF10. After cycloplegia was indued by 1.0% cyclopentolate，the refractive status was rechecked. The vector method modified by Thibos was used to analysis asitgmatism. Data only from right eye was analyzed. Results：A total of 1 882 children（response rate 98.92%）had complete data from noncycloplegic autorefraction，1 344 children（cycloplegic rate 71.41%）had complete data from cycloplegic autorefraction，so 1 344 children were included in this study. Astigmatism in 5~6 years old children before and after cycloplegia was mainly manifested as with-the-role（WTR），followed by oblique astigmatism，and then against-the-role（ATR）. Spherical equivalent（SE）values in cycloplegic refraction were significantly more hyperopic than those in noncycloplegic refraction by mean difference of（1.48 ± 1.31）diopters（P < 0.001）. The J0 values increased significantly in cycloplegic refraction except the myopic group（P=0.444），while the J45 values had no significant change in any refractive groups（all P < 0.05）. Blande-Altman plots demonstrated the agreement of astigmatism before and after cycloplegia，the 95% confidence interval of the J0 and J45 were -0.24~+0.32 and -0.15~+0.15，respectively. Conclusion：Cycloplegia has a mild effect on astigmatism，especially in the WTR and ATR astigmatisms，but do not reach clinical significant. Non-cycloplegia is still a reliable tool to measure astigmatism for preschool children.

Abstract:Objective：To quantitatively analyze the clinical application value of electronystagmography in early-stage Alzheimer’s disease（AD）by combining cerebrospinal fluid markers and cognitive neuropsychological scale. Methods：The subjects were divided into three groups：individuals with mild cognitive impairment（aMCI group），mild AD（AD group）and normal controls，with 20 cases in each group. After completing the evaluation of the cognitive scale，the three groups of patients were examined by electronystagmography and cerebrospinal fluid markers，and the results were statistically analyzed. Results：①In the saccade test，there was a significant difference in the number of accuracy abnormalities between the AD group and the other two groups（P < 0.05），and a significant difference in the latency between the control group and the other two groups（P < 0.001）. ②MOCA，SDMT，CDT，MMSE，and ADL scores were negatively correlated with saccade latency，and the attachment test scores were positively correlated with saccade latency. ③There was a significant negative correlation between the level of Aβ1-42 and the latency of saccade. Conclusion：There is eye movement damage in early-stage AD patients，which is mainly manifested by decreased accuracy and prolonged latency. Electronystagmography is helpful for early AD screening.

Abstract:Myelodysplastic syndrome （MDS） refers to a group of diseases derived from heterogeneous cloning of hematopoietic stem cells，which is characterized by cytopenia associated with ineffective hematopoiesis and a high risk of progression to acute leukemia. In the past decade，our knowledge about the genetics of MDS has been substantially improved due to the rapid development of sequencing technology，which facilitates the early diagnosis，risk stratification，interpretation of pathogenesis and the improvement of treatment of the disease. Here we review the recent literatures about the genetic abnormalities related to MDS.

Abstract:Certain chemotherapeutic drugs can elicit immunogenic cell death while inducing apoptosis of tumor cells. Generally，they have better therapeutic effects. This procedure is mainly characterized by up-regulation of certain characteristic protein molecules，such as calreticulin（CRT），high mobility group box 1（HMGB1），on the surface of apoptotic cells. These proteins can induce immature dendritic cells to maturation and present tumor antigens to cytotoxic T cells. Eventually，activated tumor-specific cytotoxic T cells kill tumor cells and ultimately achieve better therapeutic effects. The study of tumor immunogenic cell death will provide new methods and approaches for tumor treatment，and provide new ideas for tumor immunity study.