Sine oculis homeobox 1 (Six1) is an important factor for embryonic development and carcinoma malignancy. However, the localization of Six1 varies due to protein size and cell types in different organs. In this study, we focus on the expression and localization of Six1 in male reproductive organ via bioinformatics analysis and immunofluorescent detection. The potential interacted proteins with Six1 were also predicted by protein-protein interactions (PPIs) and Enrichr analysis. Bioinformatic data from The Cancer Genome Atlas and Genotype-Tissue Expression project databases showed that SIX1 was highly expressed in normal human testis, but low expressed in the testicular germ cell tumor sample. Human Protein Atlas examination verified that SIX1 level was higher in normal than that in cancer samples.The sub-localization of SIX1 in different reproductive tissues varies but specifically in the cytoplasm and membrane in testicular cells. In mouse cells, single cell RNA-sequencing data analysis indicated that Six1 expression level was higher in mouse spermatogonial stem cells (mSSCs) and differentiating spermatogonial than in other somatic cells.Immunofluorescence staining showed the cytoplasmic localization of Six1 in mouse testis and mSSCs. Further PPIs and Enrichr examination showed the potential interaction of Six1 with bone morphogenetic protein 4 (Bmp4) and catenin Beta-1 (CtnnB1) and stem cell signal pathways. Cytoplasmic localization of Six1 in male testis and mSSCs was probably associated with stem cell related proteins Bmp4 and CtnnB1 for stem cell development.

Objective:To investigate the value of ultrasound and Ki-67 for early predicting pathological complete response (pCR) of triple negative breast cancer(TNBC) after neoadjuvant chemotherapy (NAC).Methods:Retrospective analysis was performed in 190 patients with TNBC who underwent surgery after NAC treatment at the Cancer Hospital of Fudan University from January 2019 to December 2022. All patients underwent ultrasound examination before and after 2 and 4 cycles of NAC treatment. According to the operation pathological results after NAC, the patients were divided into pCR group and non-pCR group. The differences in ultrasound and Ki-67 parameters were compared between the pCR and non-pCR groups, and binary Logistic regression analysis was performed to determine the independent predictors for pCR. The ROC curve was plotted to evaluate the diagnostic efficacy.Results:Tumor maximum diameter, relative change rates of tumor maximum diameter after 2-cycle and 4-cycle NAC (ΔD2, ΔD4), relative change rate of lymph node short diameter after 2-cycle NAC (ΔS2), T-stage, N-stage and Ki-67 showed statistically significant differences between the pCR group and the non-pCR group (all P<0.05). Logistic regression analysis showed that ΔD4, T-stage, N-stage and Ki-67 were independent predictors for pCR ( OR=1.029, P=0.011; OR=0.300, P=0.009; OR=0.653, P=0.048; OR=1.028, P=0.001). The area under the curve (AUC) of pCR was 0.804 (95% CI=0.742-0.866), the sensitivity and specificity were 67.5% and 83.2% respectively. Conclusions:The combination parameters of ΔD4, T-stage, N-stage and Ki-67 have certain clinical value for predicting pCR of TNBC.

Objective:To compare the effects of two embryo culture reagents on embryonic developmental time dynamics, embryonic development potential and clinical pregnancy outcomes.Methods:The data of patients undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) were retrospectively analyzed at the Center for Reproductive Medicine of the First Affiliated Hospital of Zhengzhou University from September 2016 to May 2018. According to the different embryo culture reagents, they were divided into Vitrolife group and Cook group, comparing the embryonic time dynamics parameters, embryo development potential and clinical pregnancy outcomes of 470 patients who met the criteria. Results:There were significant differences in the kinetic parameters of embryo development between the two different embryo culture reagents. Compared with Cook group, Vitrolife group had shorter time in pronuclei appeare and disappear, and time of development to 3-cell and 8-cell embryos [tPNa: (7.12±2.71) h vs. (7.40±2.61) h, tPNf: (23.83±4.33) h vs. (24.21±4.74) h, t3: (35.75±6.03) h vs. (36.64±6.16) h, t4: (38.30±6.25) h vs. (38.92±6.06) h, t5: (47.59±7.85) h vs. (49.01±7.86) h, t6: (50.77±7.17) h vs. (52.12±6.99) h, t7: (53.05±6.31) h vs. (54.33±6.37) h, t8: (55.35±6.89) h vs. (56.31±6.41) h, all P<0.05]. There was no significant difference in cell cycle s2 and had a significant difference in cc2 [9.71±4.60) h vs.(10.33±4.28) h, P<0.001], synchronized in cell cycle in the sexual comparison. Cook group had a longer cell cycle, cell development time interval t5-t4 [(10.60±5.65) h vs. (11.20±5.90) h], t8-t4 [(18.45±5.76) h vs. (19.28±5.18) h] with statistically significant differences, all P<0.05, but there was no significant difference at t2 and t4-t2. Comparing the embryo development potential of the two embryo culture reagents, embryo utilization rate (59.9% vs. 63.9%, P=0.017) and day 3 (D3) high-quality embryo rate (65.4% vs. 69.5%, P=0.011) in Cook group were higher than those in Vitrolife group. There were no significant differences in implantation rate, embryo implantation rate, blastocyst implantation rate, fertilization rate and cleavage rate ( P>0.05). The clinical pregnancy rate, the continuous pregnancy rate (OPR), the abortion rate, the delivery rate, the live birth rate, the fetal malformation rate, the male to female ratio, the embryo pregnancy rate, and the blastocyst pregnancy rate were not statistically different ( P>0.05). Conclusion:According to the actual situation of the center, each center should find out the embryo time kinetic parameters suitable for the embryo culture reagent of the center, and establish the selection standard of the best embryo culture reagent, so as to improve the clinical pregnancy rate and live birth rate of the patients in the center.

Objective:To compare the effects of two embryo culture reagents on embryonic developmental time dynamics, embryonic development potential and clinical pregnancy outcomes.Methods:The data of patients undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) were retrospectively analyzed at the Center for Reproductive Medicine of the First Affiliated Hospital of Zhengzhou University from September 2016 to May 2018. According to the different embryo culture reagents, they were divided into Vitrolife group and Cook group, comparing the embryonic time dynamics parameters, embryo development potential and clinical pregnancy outcomes of 470 patients who met the criteria. Results:There were significant differences in the kinetic parameters of embryo development between the two different embryo culture reagents. Compared with Cook group, Vitrolife group had shorter time in pronuclei appeare and disappear, and time of development to 3-cell and 8-cell embryos [tPNa: (7.12±2.71) h vs. (7.40±2.61) h, tPNf: (23.83±4.33) h vs. (24.21±4.74) h, t3: (35.75±6.03) h vs. (36.64±6.16) h, t4: (38.30±6.25) h vs. (38.92±6.06) h, t5: (47.59±7.85) h vs. (49.01±7.86) h, t6: (50.77±7.17) h vs. (52.12±6.99) h, t7: (53.05±6.31) h vs. (54.33±6.37) h, t8: (55.35±6.89) h vs. (56.31±6.41) h, all P<0.05]. There was no significant difference in cell cycle s2 and had a significant difference in cc2 [9.71±4.60) h vs.(10.33±4.28) h, P<0.001], synchronized in cell cycle in the sexual comparison. Cook group had a longer cell cycle, cell development time interval t5-t4 [(10.60±5.65) h vs. (11.20±5.90) h], t8-t4 [(18.45±5.76) h vs. (19.28±5.18) h] with statistically significant differences, all P<0.05, but there was no significant difference at t2 and t4-t2. Comparing the embryo development potential of the two embryo culture reagents, embryo utilization rate (59.9% vs. 63.9%, P=0.017) and day 3 (D3) high-quality embryo rate (65.4% vs. 69.5%, P=0.011) in Cook group were higher than those in Vitrolife group. There were no significant differences in implantation rate, embryo implantation rate, blastocyst implantation rate, fertilization rate and cleavage rate ( P>0.05). The clinical pregnancy rate, the continuous pregnancy rate (OPR), the abortion rate, the delivery rate, the live birth rate, the fetal malformation rate, the male to female ratio, the embryo pregnancy rate, and the blastocyst pregnancy rate were not statistically different ( P>0.05). Conclusion:According to the actual situation of the center, each center should find out the embryo time kinetic parameters suitable for the embryo culture reagent of the center, and establish the selection standard of the best embryo culture reagent, so as to improve the clinical pregnancy rate and live birth rate of the patients in the center.

Objective: To investigate the application of single-molecule PCR (SM-PCR) in the detection of plasma ctDNA for the treat-ment of patients with advanced lung adenocarcinoma. Methods: In total, 30 patients diagnosed with advanced lung adenocarcinoma were enrolled between June 2017 and May 2018. ctDNA fragments of the target genes (EGFR, KRAS, BRAF, ALK, HER2, and TP53) from the blood samples were enriched by SM-PCR, and DNA libraries were prepared. Finally, a high-throughput sequencing was performed. The EGFR detection of tumor tissue samples was performed using real-time fluorescence PCR based on the amplification refractory mutation system (ARMS) and consistency in the results of EGFR mutation detection in the plasma and tissue was compared. Results:The results of both the methods were consistent (Kappa=0.867, P<0.001). The McNemar's test also indicated that the results are not statistically different (P=0.500). Conclusions: SM-PCR can be used for the detection of plasma EGFR mutations. The target detection sites are more comprehensive and multiple mutations can be detected at the same time. Results of the analysis are more precise and can be absolutely quantified.

Objective To explore the value of layer-specific strain in assessment of left ventricular function in patients with heterozygous familial hypercholesterolemia (HeFH) with normal left ventricular ejection fraction (LVEF).Methods Thirty-four patients with diagnosed HeFH and underwent transthoracic echocardiography were included as HeFH group,while 29 healthy volunteers were taken as control group.EchoPAC software was used to obtain endocardial longitudinal strain (LSendo),myocardial longitudinal strain (LSmyo) and epicardial longitudinal strain (LSepi) of the epicardial,and then statistical analysis was performed.Results LSendo and LSmyo in HeFH group were significantly lower than those in control group (P＜0.001).LSendo and LSmyo were negatively correlated with total cholesterol and low density lipoprotein cholesterol (all P＜0.05).Conclusion Layer-specific strain of left ventricular is of great value in assessing early myocardial damage in patients with HeFH.

OBJECTIVETo establish a new extrahepatic growing hepatocellular carcinoma cell line.

METHODSA specimen from extrahepatic growing hepatocellular carcinoma was cultured in vitro. Cancer cells were studied morphologically and subjected to karyotype analysis, DNA analysis, and tumor formation evaluation.

RESULTSMorphological observation and functional analysis showed that their features were similar to those of HCC. Chromosomes with a variation of 76 approximately 104 were able to secret AFP in vitro and to form bile canaliculi with microvilli.

CONCLUSIONEGHC-9901 cell line has characteristics of the extrahepatic growing hepatocellular carcinoma.

Adult ; Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Chromosome Aberrations ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Mice ; Mice, Nude ; Tumor Cells, Cultured ; alpha-Fetoproteins ; analysis

To establish a new extrahepatic growing hepatocellular carclnoma cell line. Methods: A speciment that was pathologically identified extrahepatic growing hepatocellular carcinoma were cultured in vitro. The morphology,karyotype analysis, DNA analysis, the tumor formation of heterotransplantation were observed. Results: Morphological observation and functional analysis showed that it had the comnnon features of HOC.It was found to be able to secret AFP in vitro. Conclusion: EGHC-9901 has characteristic of the extrahepatic growing hepatocellular carcinoma cell line.