Objective : To explore the role of semaphoring 6d(SEMA6D) in the malignant progression of triple-negative breast cancer(TNBC). Methods : Bioinformatics and Immunohistochemistry(IHC) were used to analyze the expression level of SEMA6D in TNBC and paracancer non-tumor tissues and its relationship with patients′ clinicopathological features. MDA-MB-231 cell line stably knocking down the expression of SEMA6D was constructed, and the effects of SEMA6D on migration and invasion of TNBC cells were investigated by Wound-healing assays and Transwell assays. cBioPortal and GEPIA2 databases were used to screen out the gene negatively associated with it, namely aurora kinase A(AURKA). Bioinformatics and IHC were used to analyze the expression level of AURKA in TNBC and paracancer non-tumor tissues and its relationship with patients' clinicopathological features. Western blot assay was used to analyze the expression of AURKA and the effect of epithelial-mesenchymal transition(EMT) makers Claudin-1, N-cadherin and Vimentin after knocking downSEMA6D. Results: Bioinformatics analysis and IHC results showed that the expression of SEMA6D in TNBC tissues was significantly lower than that in paracancer non-tumor tissues(bothP<0.05). The expression of AURKA in TNBC tissues was significantly higher than that in paracancer non-tumor tissues(bothP<0.05), SEMA6D and AURKA were significantly negatively correlated in TNBC(P<0.01). Both low expression of SEMA6D and high expression of AURKA were positively correlated with tumor size, tumor histological grade, clinical stage and lymph node metastasis in TNBC patients(allP<0.05). The knockdown ofSEMA6Dsignificantly promoted the migration and invasion ability of TNBC cells(bothP<0.01). Western blot results showed that the knockdown ofSEMA6Dupregulated AURKA expression, promoted the expression of N-cadherin and Vimentin, and inhibited the expression of Claudin-1 in tumor cells. Conclusion Down-regulation of SEMA6D expression in TNBC may be involved in the malignant progression of TNBC through up-regulation of AURKA expression and promotion of EMT.

Objective : To explore the expression of prohibitin2(PHB2) in breast cancer and its effect on the biological behaviors of tumor cells. Methods : Immunohistochemistry was used to detect the expression of PHB2 protein in breast cancer tissues and its relationship with clinicopathologic features. Breast cancer stable transient cell lines were constructed with knockdown and overexpression ofPHB2, respectively. The effects of PHB2 on cell proliferation, migration and invasion ability were detected by clone formation assay, scratch assay and Transwell assay. Western blot(WB) was used to detect the effects of PHB2 on the expression of epithelial-mesenchymal transition(EMT)-related markers, including E-cadherin, N-cadherin, Snail family transcriptional repressor 1(Snail) protein, Vimentin, and Claudin-1. The effect of PHB2 on tumorigenicityin vivowas detected by subcutaneous tumor formation assay in nude mice. Results: The result of immunohistochemical showed that PHB2 was highly expressed in breast cancer and the expression of PHB2 was significantly positive correlated with tumor size, human epidermal growth factor receptor-2(HER-2) status and proliferation index Ki-67 levels(P<0.05). Clone formation assay, scratch assay and Transwell assay revealed that knockdown ofPBH2significantly inhibited the proliferation, migration and invasion ability of breast cancer cells(P<0.01), while the overexpression ofPHB2significantly promoted cell proliferation, migration and invasion(P<0.01). The result of subcutaneous tumor formation experiment in nude mice revealed a significant decrease in tumor volume and weight in knockdownPHB2mice(P<0.000 1), whilePHB2overexpression tumors significantly increased in volume and weight(P<0.001).WB assay showed that the protein expression of epithelial marker E-cadherin increased, while the expressions of mesenchymal markers N-cadherin, Snail and Vimentin decreased significantly afterPHB2knockdown with them in control cells(P<0.01). The expression of Claudin-1 decreased, while the expressions of N-cadherin, Snail and Vimentin increased significantly inPHB2overexpression cells(P<0.05). Conclusion PHB2 is highly expressed in breast cancer and promotes multiple malignant biological behaviors in tumor cells, suggesting PHB2 may be a potential target for breast cancer diagnosis and treatment.

Objective: To simulate the tumor microenvironment though the 3D microtumor model which was constructed using droplet microfluidics.To explore its feasibility as a model for in vitro breast cancer research through3D microtumour fabrication,characterisation and sensitivity testing to chemotherapeutic drugs. Methods: Breast cancer cells were encapsulated with hydrogel shells in collagen-rich microencapsulated cores to obtain breast cancer microtumours in vitro;breast cancer microtumours were co-cultured with 3D microencapsulated endothelial cells by Transwell system.The structure and growth characteristics of the microtumours were directly observed by microscopy;the CCK-8 assay was used to detect the proliferation of the cells under different culture models and the drug sensitivity to doxorubicin;flow cytometry was used to compare the differences in apoptosis during the proliferation process;and the differences in the migratory and invasive abilities of the cells were assessed by scratch assay and Transwell assay;the expression of epithelial-mesenchymal transition-related proteins was detected by Western blot. Results: Breast cancer cells grew well in hydrogel nucleus-shell microcapsules;cell proliferation assays showed that 3D culture and 3D co-culture cells proliferated at a significantly lower rate than 2D culture;3D culture and 3D co-culture cells had enhanced migration and invasion ability and showed higher expression of EMT-related proteins compared to 2D culture;3D culture and 3D co-culture cells were significantly less sensitive to chemotherapeutic drugs compared to 2D culture.The sensitivity of 3D and 3D co-cultured cells to chemotherapeutic drugs was significantly reduced compared to 2D culture. Conclusion 3D cultures show similar morphology and biology to in vivo tumours and are more resistant to chemotherapeutic agents.

Purpose To explore the expression of autoph-agy-related 7(ATG7)in breast cancer and its effect on the breast cancer development.Methods Immunohistochemistry(IHC)was used to detect ATG7 protein expression in breast cancer tissues and the relationship between ATG7 and clinico-pathological features was analyzed.ShRNA was used to interfere with the expression of ATG7 in breast cancer cell line MCF-7.Puromycin was used to screen for stably transfected cells and Western blot was used to detect transfection efficiency.The effect of ATG7 knockdown cells on proliferation ability was de-tected by CCK8 and clone formation experiments.The effect of ATG7 knockdown cells on tumorigenicity in vivo was detected by subcutaneous tumor formation experiment in nude mice.Results IHC showed that ATG7 expression in breast cancer tissues was mainly localized in cytoplasm,and its expression was significant-ly correlated with tumor size and Ki67 expression(P＜0.05).ATG7-shRNA significantly interfered with ATG7 expression in breast cancer cells MCF-7.CCK8 and clone formation experi-ments showed that ATG7 knockdown promoted the cell prolifera-tion compared with the control group.The experiment of subcu-taneous tumor formation in nude mice showed that the tumor for-mation ability of mice was significantly increased after ATG7 knockdown compared with the control group.Conclusion ATG7 may inhibit the proliferation capacity of breast cancer and could be a potential target for breast cancer therapy.

Objective:To compare the clinical efficacy and safety of Shuo Tong ureteroscopy(ST-URS) and flexible ureteroscope(FURS)combined with holmium laser lithotripsy in the treatment of upper ureteral calculi with CT numerical value ≥ 1000 HU.Methods:A retrospective analysis of the clinical data of patients of upper ureteral calculi with CT numberical value≥1000 HU in the First Affiliated Hospital of Xiamen University was made from January 2018 to November 2020.There were 61 cases treated with ShuoTong ureteroscopy holmium laser lithotripsy (ST-URS group), including 45 males and 16 females, with 40 on the left and 21 on the right, age of(48.3±12.7) years, body mass index of(24.7±2.7)kg/m 2, the diameter of stone of(1.50±0.45)cm, and the CT numberical value of(1 288.8±179.0)(1 017-1 738)HU. There were 87 cases were treated with flexible ureteroscopy holmium laser lithotripsy (FURS group), including 58 males and 29 females, with 56 on the left and 31 on the right, age of(48.5±13.0) years, body mass index of(24.1±3.8)kg/m 2, the stone diameter of(1.45±0.40)cm, and the CT numberical value of(1 311.3±188.9)(1 009-1 817)HU. There were no significant differences in gender, age, body mass index, the location of stone, the diameter of stone and the CT numberical value of stone( P>0.05)between the two groups. For ST-URS group, a rigid ureteral channel sheath and standard mirror(F7.5/11.5)were placed under direct vision, exiting the standard mirror, leaving the channel sheath, inserting a lithotripsy mirror(F4.5/6.5)and a holmium laser[Power: 8-30 W(0.4-1.0 J/20-30 Hz)], and withdrawing the stone fragments after crushing the stone by "nibbling method" . For FURS group, a hard ureteroscope(F8/9.8)was used to explore the lesion side of the ureter, inserting a guide wire and placing a soft ureteral sheath, then inserting a flexible ureteroscope(F8)for holmium laser lithotripsy, and useing a stone basket to remove larger stone fragments. Ureteral stent was routinely indwelled after the operation. On the day 1 and 1 month after the operation, imaging examinations were performed to evaluate the stone-free rate. No residual stones or the diameter of stone was ≤0.4 cm and no urinary tract infection or any symptoms were defined as stone free. The operation time, blood loss, success rate of stage Ⅰ ureteral access sheath placement, incidence of postoperative complications, stone-free rate(SFR) at 1 day after operation, SFR at 1 month after operation, postoperative hospital stay and hospitalization costs were compared between the two groups. According to the size of calculi, the 2 groups were divided into 2 subgroups(≥1.5 cm and <1.5 cm)in order to make further analysis. The operation time, stone-free rate(SFR) at day 1 after operation and SFR at 1 month after operation were compared between the two groups. Results:The operation time of the ST-URS group was shorter than the FURS group(40.10 min vs. 49.43 min, P=0.020), and the incidence of postoperative complications was lower than the FURS group[3.28%(2/61)vs. 13.79%(12/87), P=0.031]. The SFR at day 1 after operation was significantly higher than the FURS group[60.7%(37/61)vs. 25.3%(22/87), P<0.01], and the hospitalization cost was lower than that of the FURS group(27 686 yuan vs. 32 281 yuan, P<0.010). There were no significant differences in the blood loss[(4.92±9.51)ml vs.(3.95±6.04)ml, P=0.452], success rate of stageⅠureteral access sheath placement[ 96.7%(59/61)vs. 96.6%(84/87), P=1.000], SFR at 1 month after operation[81.97%(50/61) vs. 75.86%(66/87), P=0.375] and postoperative hospital stay[(2.5±1.4)d vs.(2.4±0.8)d, P=0.543] between the two groups. When the size of calculi was ≥1.5cm, the operation time of the ST-URS group was shorter than the FURS group (43.67 min vs 55.00 min), the SFR at 1 day after operation was higher than the FURS group[40.00%(12/30)vs. 9.38%(3/32)], and the above differences are all statistically significant ( P<0.05). Conclusions:Compared with the FURS, for the treatment of upper ureteral calculi with CT numerical value ≥1000 HU, the ST-URS has shorter in operative time, lower in hospitalization cost and incidence of postoperative complications and higher SFR at day 1 after operation. The ST-URS is a safe and effective surgical technique, which is superior in the treatment of larger(≥1.5 cm) stones.

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults and is poorly controlled. Previous studies have shown that both macrophages and angiogenesis play significant roles in GBM progression, and co-targeting of CSF1R and VEGFR is likely to be an effective strategy for GBM treatment. Therefore, this study developed a novel and selective inhibitor of CSF1R and VEGFR, SYHA1813, possessing potent antitumor activity against GBM. SYHA1813 inhibited VEGFR and CSF1R kinase activities with high potency and selectivity and thus blocked the cell viability of HUVECs and macrophages and exhibited anti-angiogenetic effects both in vitro and in vivo. SYHA1813 also displayed potent in vivo antitumor activity against GBM in immune-competent and immune-deficient mouse models, including temozolomide (TMZ) insensitive tumors. Notably, SYHA1813 could penetrate the blood-brain barrier (BBB) and prolong the survival time of mice bearing intracranial GBM xenografts. Moreover, SYHA1813 treatment resulted in a synergistic antitumor efficacy in combination with the PD-1 antibody. As a clinical proof of concept, SYHA1813 achieved confirmed responses in patients with recurrent GBM in an ongoing first-in-human phase I trial. The data of this study support the rationale for an ongoing phase I clinical study (ChiCTR2100045380).

Objective:To explore the clinical value of urine semi-quantitative colorimetry by sodium dithionite reduction method in the diagnosis and treatment of diquat poisoning.Methods:The data of 49 patients with acute diquat poisoning treated in the First Affiliated Hospital of Nanjing Medical University from December 3, 2020 to November 23, 2022 were retrospectively analyzed, the correlation between urine colorimetric results and plasma diquat concentration was observed, and the predictive value of urine colorimetric results for target organ damage and prognosis were evaluated.Results:There was a significant correlation between urine colorimetric results and plasma diquat concentration, the correlation coefficient was r=0.89, P <0.01. The cut-off value of urine colorimetry for the predicting the damage of gastrointestinal tract, liver, kidney, and central nervous system injury were 2.5, 3.5, 3.5, 5.5, respectively; in which the urine colorimetric results showed the highest sensitivity in predicting digestive tract injury [ AUC 0.93 (95% CI:0.89-1.00)]. The cut-off value of urine colorimetry for the prognosis of death was 4.5, the positive predictive value was 64.2%, and the negative predictive value was 95.2%. Conclusions:The urine semi-quantitative method can be used for rapid prediction of the plasma diquat concentration range on admission. The urine colorimetry results can also effectively predict the occurrence of organ injury and clinical outcome related to diquat poisoning, which provides evidence for the clinical diagnosis and therapy.

Purpose To investigate the expression of PD-1,PD-L1,CD3 and CD8 in the immune microenvironment of non-small cell lung cancer(NSCLC)and their clinical signifi-cance.Methods The clinical data of 39 patients with NSCLC brain metastasis(BM)were collected.The expression of PD-1,PD-L1,CD3 and CD8 in the tumor and stroma of BM was detec-ted using an immunofluorescence-based tissue microenvironment analysis panel.Targeted sequencing was carried out to catalog cancer-related genes.The clinical pathological features were an-alyzed with review of relevant literature.Results Thity-nine patients with NSCLC presented with tumor-infiltrating lympho-cytes(TIL)in different degree.CD3+TIL(P=0.000 7)and CD8+TIL(P=0.0006)were more prominent in the tumor stroma,and the positive PD-L1 expression was significantly higher in the interstitial tissues of tumor(P=0.025 8).Com-pared with the whole wild-type driver gene cohort,the expres-sion of PD-L1 in the stroma of BM was significantly increased in the EGFR mutation cohort(P=0.039).Patients with high in-filtration of stroma CD8+TIL had longer median overall survival than those with low infiltration(16 months vs 6 months,P=0.032);PD-L1-positive patients(36 months)were longer sur-vival time than PD-L1-negative patients(5.5 months,P=0.056).Compared with lung primary lesions,the variant allele frequencies(VAFs)in BM generally increased,and samples with higher VAFs corresponded to higher expression of PD-1,PD-L1,CD3 and CD8.Conclusion The TIL infiltration is most prominent in the stroma of NSCLC BM.The EGFR muta-tion of a tumor might affect the immune microenvironment of me-tastases;PD-L1 expression and TIL infiltration were correlated with overall survival.

Objective To investigate the influencing factors for the prognosis of adult patients with chronic drug-induced liver injury (DILI). Methods A total of 255 patients who were diagnosed with chronic DILI by liver biopsy in The Fifth Medical Center of Chinese PLA General Hospital from January 2014 to December 2018 were enrolled, and according to the liver function after 2 years, they were divided into non-recovery group and recovery group. The two groups were analyzed in terms of the clinical data including age, sex, body mass index, types of drugs used, type of DILI injury, severity of DILI injury, underlying diseases, laboratory markers, liver histology, and 2-year prognosis. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. Univariate and multivariate logistic regression analyses were used to investigate the independent risk factors for the prognosis of chronic DILI. Results After 2 years of follow-up, 195 patients (76.5%) achieved the recovery of liver function, while 60 patients (23.5%) did not achieve such recovery. There were significant differences between the two groups in the type of DILI injury ( P =0.028), the proportion of patients with diabetes ( P =0.048), and the degree of liver fibrosis ( P < 0.001), and compared with the recovery group, the non-recovery group had significantly higher levels of baseline white blood cell count, platelet count (PLT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase, and total bile acid and a significantly lower level of cholinesterase (ChE) (all P < 0.05). The baseline characteristics were included in the univariate logistic regression analysis, and the results showed that PLT, ALT, AST, ChE, and fibrosis degree were significantly associated with the prognosis of chronic DILI (all P < 0.05). The multivariate logistic regression analysis of the above variables showed that PLT < 100×10 9 /L (odds ratio [ OR ]=3.592, 95% confidence interval [ CI ]: 1.128-11.438, P =0.003) and ALT > 2×upper limit of normal (ULN) ( OR =3.080, 95% CI : 1.331-7.127, P =0.009) were independent risk factors for the prognosis of chronic DILI. Conclusion When patients meet the diagnostic criteria for chronic DILI, the independent risk factors PLT < 100×10 9 /L and ALT > 2×ULN may be used to screen out the patients who are more likely to have poor prognosis.

Objective:To establish surgical difficulty scoring system of partial nephrectomy based on holographic imaging and explore its application value in partial nephrectomy.Methods:A total of 184 patients including 110 males and 74 females with renal tumors diagnosed as stage cT 1 to cT 2 before surgery in the First Affiliated Hospital of Xiamen University from October 2019 to January 2022 were included. Among the 184 patients, 141 patients were treated with partial nephrectomy and 43 patients were treated with radical nephrectomy (3 partial nephrectomies were finally changed to radical nephrectomies due to vascular and tumor location). 60 patients had hypertension. 24 patients had diabetes. 7 patients had hyperuricemia. The median age was 55(47, 62) years. The median BMI was 23.7(21.8, 26.4) kg/m 2. The median maximum tumor diameter was 3.9(2.9, 5.2) cm. The median preoperative eGFR was 99.7(83.4, 114.2) ml/(min·1.73m 2). The median R. E.N.A.L. score was 8(6, 9). The median PADUA score was 9(8, 10). 153 patients were diagnosed as stage cT 1 and 31 patients were diagnosed as stage cT 2. The hologram reconstruction was performed according to preoperative CT or MRI examination. The maximum diameter of the tumor in the kidney (D), the compression degree of the renal segmental vessels by tumor(C), the area of the renal sinus occupied by tumor(O) and the mass of exophytic rate(M) were comprehensively considered and finally constituted the difficulty scoring system named DCOM score for partial nephrectomy. The DCOM score divided the complexity of tumor surgery into mild (4-6 points), moderate (7-8 points) and high (≥ 9 points). Meanwhile, the MIC (surgical margins are negative, WIT is <20 min, and no major complications)was used to evaluate the overall surgical effect. The DCOM, R. E.N.A.L. and PADUA scores were performed on all patients and compared with each other to evaluate the surgical effect of DCOM score in partial nephrectomy. Results:All surgeries in this study were successfully completed, including 141 partial nephrectomies and 43 radical nephrectomies. The DCOM score was 10(9, 11) for radical nephrectomy and 6(5, 8) for partial nephrectomy, and the difference was statistically significant ( P=0.001). There were 23 patients (37.7%) in highly complex group, 39 patients (88.6%) in moderately complex group and 79 patients (100.0%) in mildly complex group underwent partial nephrectomy, respectively. According to multifactorial analysis, patients in highly and moderately complex group of DCOM score had 8.88 times ( P=0.001) and 1.76 times ( P=0.005) less reach MIC than those in mildly complex group, respectively. Patients in highly and moderately complex group of PADUA score had 4.86 times ( P=0.005)and 3.41 times ( P=0.006)less reach MIC than patients in mildly complex group of DCOM score, respectively. What’s more, patients in moderately complex group of R. E.N.A.L. score had 3.11 times ( P=0.003) less reach MIC than patients in mildly complex group of DCOM score. In the ROC curves to predict MIC achievement, the AUC values of R. E.N.A.L., PADUA and DCOM scores were 0.657, 0.655 and 0.746, respectively. Comparing:R. E.N.A.L. score with DCOM score, the AUC value was statistically significant ( P=0.025). Conclusions The surgical difficulty scoring system (DCOM score) based on holographic imaging can predict the outcome of partial nephrectomy, but further verification is needed.