BACKGROUND: P16 inactivation is frequently accompanied by telomerase reverse transcriptase ( TERT ) amplification in human cancer genomes. P16 inactivation by DNA methylation often occurs automatically during immortalization of normal cells by TERT . However, direct evidence remains to be obtained to support the causal effect of epigenetic changes, such as P16 methylation, on cancer development. This study aimed to provide experimental evidence that P16 methylation directly drives cancer development. METHODS: A zinc finger protein-based P16 -specific DNA methyltransferase (P16-Dnmt) vector containing a "Tet-On" switch was used to induce extensive methylation of P16 CpG islands in normal human fibroblast CCD-18Co cells. Battery assays were used to evaluate cell immortalization and transformation throughout their lifespan. Cell subcloning and DNA barcoding were used to track the diversity of cell evolution. RESULTS: Leaking P16-Dnmt expression (without doxycycline-induction) could specifically inactivate P16 expression by DNA methylation. P16 methylation only promoted proliferation and prolonged lifespan but did not induce immortalization of CCD-18Co cells. Notably, cell immortalization, loss of contact inhibition, and anchorage-independent growth were always prevalent in P16-Dnmt&TERT cells, indicating cell transformation. In contrast, almost all TERT cells died in the replicative crisis. Only a few TERT cells recovered from the crisis, in which spontaneous P16 inactivation by DNA methylation occurred. Furthermore, the subclone formation capacity of P16-Dnmt&TERT cells was two-fold that of TERT cells. DNA barcoding analysis showed that the diversity of the P16-Dnmt&TERT cell population was much greater than that of the TERT cell population. CONCLUSION P16 methylation drives TERT -mediated immortalization and transformation of normal human cells that may contribute to cancer development.
Humans ; Telomerase/genetics* ; DNA Methylation/physiology* ; Fibroblasts/cytology* ; Cyclin-Dependent Kinase Inhibitor p16/metabolism* ; Cell Line ; Cell Transformation, Neoplastic/genetics*

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To explore the application effect of pelvic floor outpatient intervention during pregnancy on vaginal delivery of full-term primiparous women.Methods:Single birth primipara who were willing to establish a card and have a desire to give birth naturally in the First People's Hospital of Longquanyi District from January 2021 to December 2021 were divided into odd and even number groups according to the order of filing:the odd group received routine obstetric outpatient management and pelvic floor outpatient intervention(Kegel exercise joint perineum massage)in 200 cases(research group),and even number group received routine obstetric outpatient management in 200 cases(control group).The incidence of vaginitis,cesarean section rate,vaginal delivery rate,obstetric laceration,vaginal midwifery rate,perineal incision,delivery outcome and the length of perineal body and reproductive hiatus before and 42 days after delivery,vaginal width andother vaginal delivery complications(urina-ry retention and puerperal hemorrhoids)were compared between the two groups.Results:87 cases were includ-ed in the final study group,while 104 cases were included in the control group.The perineal incision rate,perineal integrity rate,vaginal midwifery rate,l degree and Ⅱ degree perineal laceration rate,postpartum hemorrhage in the study group were better than those in the control group(P＜0.05).The length of reproductive hiatus 42 days after delivery in the study group was shorter compared with that in the control group(P＜0.05).There were no signifi-cant differences in the incidence of vaginitis,cesarean section rate,time of the second stage of labor,vaginal wall laceration rate,Apgar score of newborns neonatal birth trauma rate,urinary retention,puerperal hemorrhoids,post-partum vaginal width and perineal body length between the two groups(P＞0.05).Conclusions:Pelvic floor outpa-tient intervention during pregnancy can reduce the harm caused by vaginal delivery in full-term primiparas to a cer-tain extent,and does not increase the incidence of neonatal delivery complications,it is worthy of clinical promotion.

Objective:To analyze the use of policy tools for rural-oriented tuition-waived medical student training policy and to provide relevant suggestions for the continuous promotion of the policy.Methods:With "rural-oriented tuition-waived medical students" as the key word, the policy texts were collected and screened from government portals. Using ROTHWELL disaggregated method to build the rural order directional medical students training policy analysis framework, applying Excel 2019 software for classification and coding of policy texts.Results:A total of 13 rural-oriented medical student training policy texts were screened and obtained. The X dimension of the policy analysis framework for rural order-oriented medical student training included three policy tools, namely, supply, environment and demand, and the Y dimension included three policy objectives, namely, available, usable, and retained. In X dimension, environmental policy tools were most frequently used. In Y dimension, the "retained" target had the highest frequency of use.Conclusion:There were differences in the frequency of using policy tools for targeted medical student cultivation in different policies. The frequency of using environmental tools is higher, which highlighted the attention of the state to medical and health services. The internal structure of policy tools is unbalanced, so the configuration of supply-oriented policy tools should be optimized, and the construction of demand-oriented policy tools should be emphasized. It is suggested to continuously optimize the combination of policy tools, improve the compatibility between policy tools and rural order-oriented medical student training, and pay attention to the sustainability of policy tools.