Objective:To investigate the effects of different semen sample collection methods on sperm DNA fragmentation index (DFI) test results, then to evaluate the accuracy of the current semen sample collection method in the assessment of male fertility.Methods:In this study, 50 semen sample obtained on the day of oocyte retrieval from patients undergoing in vitro fertilization at the Reproductive Medical Center in Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region from August 2021 to January 2022 were collected. For each semen, a small amount of samples were collected in three different retention methods for routine semen and sperm DFI testing. Three different ways of retaining samples were as follows: group A, after mixing of the semen, 50 μL sample was directly collected; group B, after density gradient centrifugation, 50 μL sample was collected at the interface between semen and gradient fluid; group C, after density gradient centrifugation, the sperm pellet was upstream, then 50 μL sample was collected from upstream liquid. After semen treatment, routine semen testing and sperm DFI testing were performed. Pearson was used to analyze the correlation between DFI and the percentage of immobile sperm and the percentage of forward sperm movement. Results:The sperm motility rate of group C [(96.83±2.28)%] was significantly higher than that of group A [(57.16±11.28)%, P<0.001] and group B [(22.54±9.35)%, P<0.001], and there was a statistical difference among the three groups. The immotile sperm rate of group B sample was (77.46±9.35)%, which was significantly higher than that of samples from group C [(3.14±2.31)%, P<0.001] and group A [(42.83±11.28)%, P<0.001]. There was also a statistical difference in DFI among the three groups ( P<0.001). The DFI of group B [37.18% (30.41%, 47.80%)] was significantly higher than that of group A [22.00% (14.75%, 29.25%), P<0.001] and group C [0.78% (0.00%, 2.07%), P<0.001]. Pearson analysis results showed that the DFI of group A and group B was positively correlated with the percentage of immobile sperm ( r=0.304, P=0.032; r=0.612, P<0.001), while the DFI of group B was negatively correlated with the percentage of sperm forward movement ( r=-0.517, P<0.001). Conclusion:For the same semen, the DFI of immotile sperm was significantly higher than that of motile sperm. Therefore, due to the interference of immotile sperm, the DFI value by the current sample retention method cannot accurately reflect the DNA status of active sperm participating in fertilization. This suggests that the samples used for DFI testing should be collected from motile sperm collected by gradient centrifugation, upstream or other methods, which can more accurately assess male fertility.

Objective:To investigate the effects of different semen sample collection methods on sperm DNA fragmentation index (DFI) test results, then to evaluate the accuracy of the current semen sample collection method in the assessment of male fertility.Methods:In this study, 50 semen sample obtained on the day of oocyte retrieval from patients undergoing in vitro fertilization at the Reproductive Medical Center in Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region from August 2021 to January 2022 were collected. For each semen, a small amount of samples were collected in three different retention methods for routine semen and sperm DFI testing. Three different ways of retaining samples were as follows: group A, after mixing of the semen, 50 μL sample was directly collected; group B, after density gradient centrifugation, 50 μL sample was collected at the interface between semen and gradient fluid; group C, after density gradient centrifugation, the sperm pellet was upstream, then 50 μL sample was collected from upstream liquid. After semen treatment, routine semen testing and sperm DFI testing were performed. Pearson was used to analyze the correlation between DFI and the percentage of immobile sperm and the percentage of forward sperm movement. Results:The sperm motility rate of group C [(96.83±2.28)%] was significantly higher than that of group A [(57.16±11.28)%, P<0.001] and group B [(22.54±9.35)%, P<0.001], and there was a statistical difference among the three groups. The immotile sperm rate of group B sample was (77.46±9.35)%, which was significantly higher than that of samples from group C [(3.14±2.31)%, P<0.001] and group A [(42.83±11.28)%, P<0.001]. There was also a statistical difference in DFI among the three groups ( P<0.001). The DFI of group B [37.18% (30.41%, 47.80%)] was significantly higher than that of group A [22.00% (14.75%, 29.25%), P<0.001] and group C [0.78% (0.00%, 2.07%), P<0.001]. Pearson analysis results showed that the DFI of group A and group B was positively correlated with the percentage of immobile sperm ( r=0.304, P=0.032; r=0.612, P<0.001), while the DFI of group B was negatively correlated with the percentage of sperm forward movement ( r=-0.517, P<0.001). Conclusion:For the same semen, the DFI of immotile sperm was significantly higher than that of motile sperm. Therefore, due to the interference of immotile sperm, the DFI value by the current sample retention method cannot accurately reflect the DNA status of active sperm participating in fertilization. This suggests that the samples used for DFI testing should be collected from motile sperm collected by gradient centrifugation, upstream or other methods, which can more accurately assess male fertility.

Innovative and practical platform safety management is the foundation of the innovation and entrepreneurship for undergraduate students in universities and colleges, which plays a critical role in maintaining the stability of the university and society, and ensuring the safety of teachers and students. In this present paper, the complex situation and problems of innovative and practical platform safety management, such as the imperfect regulations, weak atmosphere, incomplete facilities, unprofessional management, etc., are discussed under the background of "innovation and entrepreneurship" in China. With the combination of the above problems and the actual situation of Chengdu Medical College, the constructions of the regulations, such as access regulation, certification regulation, check regulation, rewards and punishment regulation, responsibility regulation, and a series of standard operating procedures (SOP) have also been carried out. Furthermore, adequate safety education and proper safety drills have been performed during the whole process of safety management. In addition, a diverse and professional management team with putting more emphasis on the development of safety facilities has been formed. In conclusions, the practice and exploration on the safety management of undergraduate students' innovative and practical platform in this paper may provide a theoretical basis and practical value to optimize the safety management in local universities and colleges under the background of "innovation and entrepreneurship" in China.

Objective: To recombine the induced pluripotent stem cells (iPSC) derived from the urine of septic encephalopathy (SE) patients, and provided a specificity cell model to explore the mechanism of the neuronal damage and treatment for SE patients. Methods: Urine of SE patient was collected, and tubular epithelial cells were isolated and cultured from the urine. iPSC were derived from SE patient by introducing 4 transcription factors OCT4, Klf4, Sox2, c-Myc (OKSM) into patient-specific urine cells by Millipore's Human STEMCCA^TM Constitutive Polycistronic (OKSM) Lentivirus Kit. Colony morphology, alkaline phosphatase (AKP) activity, immunofluorescence staining, quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and differentiation ability were used to identify the pluripetency of these iPSC lines. In addition, neurons were derived from these iPSC by inhibiting transforming growth factor-β (TGF-β) pathway. Results: The SE-iPSC exhibited morphological and growth characteristics of human embryonic stem cell (hES), showed positivity for AKP by histochemical staining, and expressed embryonic stem cell (ESC) marker genes. There was a significant statistical difference in ESC-marker mRNA expression between the SE-iPSC and the urine cells [NANOG mRNA (2^-ΔΔCt): 1.153±0.142 vs. 0.126±0.024, t = -10.688; REX1 mRNA (2^-ΔΔCt): 1.419±0.206 vs. 0.103±0.066, t = -14.245; OCT4 mRNA (2^-ΔΔCt): 1.233±0.176 vs. 0.201±0.022, t = -9.028; Sox2 mRNA (2^-ΔΔCt): 1.334±0.119 vs. 0.159±0.017, t = -12.653, all P < 0.01]. Subcutaneous injection of iPSC into NOD-SCID mice resulted in teratomas containing tissues from all the 3 germ layers. Furthermore, neurons were successfully induced from SE-iPSC. Conclusion The SE patient-specific iPSC could be generated from urine cells and differentiated into neurons, furthermore, the SE-iPSC cell line can be used as models for further elucidating the cellular pathology and developing therapeutic strategies for SE.

OBJECTIVE: To recombine the induced pluripotent stem cells (iPSC) derived from the urine of septic encephalopathy (SE) patients, and provided a specificity cell model to explore the mechanism of the neuronal damage and treatment for SE patients. METHODS: Urine of SE patient was collected, and tubular epithelial cells were isolated and cultured from the urine. iPSC were derived from SE patient by introducing 4 transcription factors OCT4, Klf4, Sox2, c-Myc (OKSM) into patient-specific urine cells by Millipore's Human STEMCCA^TM Constitutive Polycistronic (OKSM) Lentivirus Kit. Colony morphology, alkaline phosphatase (AKP) activity, immunofluorescence staining, quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and differentiation ability were used to identify the pluripetency of these iPSC lines. In addition, neurons were derived from these iPSC by inhibiting transforming growth factor-β (TGF-β) pathway. RESULTS: The SE-iPSC exhibited morphological and growth characteristics of human embryonic stem cell (hES), showed positivity for AKP by histochemical staining, and expressed embryonic stem cell (ESC) marker genes. There was a significant statistical difference in ESC-marker mRNA expression between the SE-iPSC and the urine cells [NANOG mRNA (2^-ΔΔCt): 1.153±0.142 vs. 0.126±0.024, t = -10.688; REX1 mRNA (2^-ΔΔCt): 1.419±0.206 vs. 0.103±0.066, t = -14.245; OCT4 mRNA (2^-ΔΔCt): 1.233±0.176 vs. 0.201±0.022, t = -9.028; Sox2 mRNA (2^-ΔΔCt): 1.334±0.119 vs. 0.159±0.017, t = -12.653, all P < 0.01]. Subcutaneous injection of iPSC into NOD-SCID mice resulted in teratomas containing tissues from all the 3 germ layers. Furthermore, neurons were successfully induced from SE-iPSC. CONCLUSIONS The SE patient-specific iPSC could be generated from urine cells and differentiated into neurons, furthermore, the SE-iPSC cell line can be used as models for further elucidating the cellular pathology and developing therapeutic strategies for SE.
Animals ; Brain Diseases ; Cells, Cultured ; Fibroblasts ; Humans ; Kruppel-Like Factor 4 ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Sepsis

Based on MOOC teaching mode and the combination of flipped classroom teaching theory,the role of internship practice is integrated into medical education and the combination of MOOC teaching and traditional teaching mode in medical humanities curriculum is explored in practice applications.This paper analyzed the cur-rent situation of medical humanities curriculum teaching operation under current situation; based on the teaching concept of MOOC,introduced the reform of teaching form and teaching method in blending teaching mode,and ex-plored the practical application and practical thinking of blending teaching mode.This mode not only plays a lead-ing role of teachers teaching,but also fully embodies the subjective status of students'learning,exercises the students'ability of thinking in practice at the same time,and is an effective mode to improve medical humanities curriculum teaching.

Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.

Pulmonary benign metastasizing leiomyoma (PBML) is a rare and otfen to be misdiagnosed disease. In this study, we want to investigate the diseased of the clinicopathological characteristics, diagnosis, treatment and prognosis. hTe retrospective analysis were performed on the clinicopathological data of 5 patients with PBML. All 5 cases were female, mean age 46.8 years old, and were found with single nodule. One case was found with letf kidney metastasis. Surgical procedures in-cludes:VATS biopsy (3 cases), resection of superior lobe of letf lung (1 case), resection of superior lobe of right lung and wedge resection of middle and inferior lobe of right lung (1 case). hTe residual nodules in lung were stable in all cases with followup 3-48 mo. PBML is dominated occurring in females with history of uterine leiomyoma, preferential to metastasize to lung, and surgery is the primary therapy. Since it is hormone dependent, hormonal therapy may be suggested in these patients.

OBJECTIVETo explore the effects of CXCR7 knock-down by CXCR7-shRNA lentiviral vector on the proliferation and invasion of human hepatoma carcinoma cells in vitro.

METHODSCXCR7-shRNA lentiviral vector was transfected into hepatocellular carcinoma HCCLM3 cells. The changes in mRNA and protein expression of CXCR7 in the transfected cells were investigated using real-time PCR and Western blotting, respectively, and MTT assay was employed to assess the cell proliferation changes. In vitro adhesion assay and transwell chamber test were used to observe the adhesion and invasiveness of HCCLM3 cells, respectively.

RESULTSTransfection of HCCLM3 cells with CXCR7-shRNA lentiviral vector resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P<0.01) and obvious suppression of the cell proliferative activity (P<0.05). CXCR7-shRNA also significantly suppressed the invasiveness and adhesion of HCCLM3 cells (P<0.01).

CONCLUSIONCXCR7 knock-down can significantly inhibit the proliferation and invasiveness of human hepatoma carcinoma cells in vitro, suggesting the value of CXCR7 as a potential target for hepatoma carcinoma therapy.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Liver Neoplasms ; genetics ; pathology ; RNA, Small Interfering ; genetics ; Receptors, CXCR ; genetics ; Transfection

With the rapid development of medical genetics,online Medelian inheritance in man (OMIM) manifests a more and more important role in medical genetics teaching.Using the educational form combining ‘ classroom teaching,review writing and seminar’,‘ Query and use of OMIM ’was introduced into the education of medical genetics.Reality practice revealed that this educational practice maintained advanced and timely status of knowledge and deeply activated self-studying and independent thinking ability of students.