Background and purpose: Pancreatic intraepithelial neoplasia (PanIN) is thought to be a precursor lesion of infiltrating pancreatic ductal adenocarcinoma. The mutation of the phenotypic impact of K-ras G12D alone, silencing of p53 and p16 could promote this process. The role of Smad4 in this progression was poorly understood. In the present study, we investigated the role of Smad4 in the development of pancreatic tumor, based on PanIN cell line from mice with K-ras G12D mutation in order to investigate the effect of Smad4 silencing on PanIN cells in the development and malignant transformation in nude mice. Methods: Smad4 knock-down PanIN cells (PanIN-S) were established by stable transfeetion with lentiviral-mediated Smad4 RNA interference (RNAi). In xenograft model experiments, BALB/c nude mice were randomly divided into 2 groups (5 mice per group) implanted with PanIN or PanlN-S cells subcutaneously. Two weeks after tumor cells inoculation, tumor volume and weight were estimated. PCNA staining was used to evaluate cell proliferation and CD31 polyclonal antibody was used to assess micro-vessel density (MVD) in tumors. Results: Effect of siRNA of Smad4 gene in PanlN cells was confirmed by RT-PCR and Western blot. Compared with PanlN groups, there was a dramatic increase in tumor volume and weight in PanIN-S groups (P<0.05). Furthermore, immunohistochemical analysis of the harvested tumors suggested that Smad4 silencing was associated with 'increased tumor cell proliferation (PCNA reactivity) and angiogenesis (micro-vessel density, MVD). The percentage of PCNA-positive cells in the PanlN-S groups were significantly increased than PanIN groups (P<0.05). CD31 staining revealed a significant increase in the PanlN-S groups compared to the PanlN groups (P<0.05). Conclusion: Silencing of Smad4 in PanlN cells with endogenous expression of K-ras G12D, enhanced progression to invasive adenocarcinomas. Cell proliferation and vascularization may be its important mechanisms.

Objective To explore the effects of cranioplasty onneurological functionin patients based on the cerebral CT perfusion technique. Methods Twenty cases of patients receiving cranioplasty were rerecorded during the study period,and they wererespectively scanned by CT perfusion within 72 hours before and 2 weeks after the cranioplasty. Meanwhile,the neurological function was evaluated by neurological function scale. Results The difference of cerebral blood flow before and after cranioplasty was statistically significant(P<0.05), whereas the difference of cerebral blood volume,transit time to the peak and mean transit time was not statistically significant(P>0.05).Correlation analysis showed that the preoperativedifference ratio of thecerebral blood infusio-nis not correlated with the neurological function score(P > 0.05). The changes of preoperative and postoperative difference rateof the cerebral blood infusionwas correlated with the functional independence measure(P < 0.05), whereas not with mini-mental state examination(P > 0.05). Conclusions The neurological function of the patients after cranioplasty may be improved.This improvement may benefit from the improvement of cerebral blood flow after cranioplasty.

Oral and maxillofacial malignant tumors threaten the life and health of patients, and seriously affect their swallowing, language function and face. 125I seeds brachytherapy for oral and maxillofacial malignant tumors has been widely concerned and studied because of its advantages such as less surgical trauma, large and uniform dose distribution in the target tissue, little damage to the surrounding normal tissue, and reducing radiation exposure of medical staff. Low-dose brachytherapy with 125I seeds can effectively reduce the tumor volume and prolong the survival time of patients. This article reviews the clinical application of 125I seeds brachytherapy in oral and maxillofacial malignant tumors.

Objective To investigate the neuroprotective effect and mechanism of liraglutide on diabetic rats. Methods 24 healthy male SPF Goto-Kakizaki (GK) rats with random blood glucose greater than 11.1 mmol/L were selected as the experimental group, and randomly divided into diabetes mellitus group ( n＝12) and liraglutide group (n＝12). Ten healthy male SPF Wistar rats with the same age and weight as GK rats were selected as normal control group. After adaptively feeded for 2 weeks, the liraglutide group was given liraglutide (400 μg·kg－1·d－1, subcutaneous injection), while the control group and diabetes mellitus group were given the same volume of saline, and continued to be administered for 8 weeks. After 10 weeks, data and biochemical indicators were recorded. Effects of liraglutide on learning and memory in diabetes mellitus rats were detected by Morris water maze test. HE staining observed the hippocampal neurons morphology. Western blotting method detected the expression of p- IκB kinase (IKK) β, p-NF-κB, NF-κB, Klotho, and PRX2 in hippocampus. Results Morris water maze test showed that liraglutide can improve the spatial learning and memory ability of diabetes mellitus rats. HE staining showed that liraglutide significantly reduced the pathological damage of hippocampal neurons of diabetes mellitus rats. Western blotting showed that liraglutide inhibited NF-κB signaling pathway in hippocampus of diabetes mellitus rats. The expression of Klotho protein in hippocampus of diabetes mellitus group was significantly lower than that of control group, while the expression of PRX2 protein was higher than control group (t＝8.298,－7.398,all P<0.01). The expression of Klotho and PRX2 protein in hippocampus of liraglutide group were higher than diabetes mellitus group (t＝－13.059, 14.113, all P<0.01). The expression of Klotho protein of liraglutide group was similar to that of control group ( t ＝ －1. 137, P>0. 05 ). The expression of PRX2 protein was significantly higher than control group (t＝－28.055, P<0.01). Conclusions Liraglutide may enhance the expression of antioxidant stress protein including Klotho and PRX2, by inhibiting NF-κB signaling pathway in hippocampus of diabetes mellitus rats, reduced oxidative stress and improved the injury of hippocampal neuronal in diabetes mellitus rats, which seems to play a neuroprotective effect, to prevent and delay the occurrence of diabetic encephalopathy.

Objective: To explore physical health status and influencing factors of preschool children in Tibet, so as to provide a scientific reference and theoretical basis for the healthy development of physical fitness. Methods: Data were collected from children (3-6 years old) of Xizang national physical fitness monitoring, and a total of 1 521 preschool children were recruited who received questionnaire survey and physical fitness test. Independent sample t-test and one-way ANOVA were used to compare the differences in physical fitness indexes of preschool children in different groups. Chi square test and binary Logistic regression model were used to analyze the factors affecting physical health of preschool children. Results: The total qualified rate of physical fitness was 79.75%, among which the excellent rate was 3.16%, the good rate was 15.12%, the qualified rate was 61.47% and the unqualified was 20.25%. From the perspective of BMI, the excellent physical fitness rate (3.74%) and good physical fitness rate ( 17.47% ) were highest in healthy weight preschool children, and the qualified physical fitness rate of overweight preschool children (69.03%) was higher than that of obese (55.88%) and healthy weight preschool children (60.68%)( χ 2=53.56, P <0.01). From the perspective of ethnic, Tibetan children s physique proficiency (3.69%), good rate ( 17.13% ) than with the elevation of the Han nationality (0.74%, 5.88%), fraction defective (16.97%), lower than that of Han nationality (35.29%) ( χ 2=53.71, P <0.01); The results of chest circumference, skinfold thickness, body fat percentage, tennis throw distance, continuous jump of both feet, sitting forward bend and balance beam walking of Tibetan children were higher than those of Han children, and the results of quiet heart rate and standing long jump were lower than those of Han children ( t = 2.72 , 10.95, 9.66, 3.68, 3.88, 8.04, 3.56, 8.70, -4.39 , -4.40, P <0.01). Binary Logistic regression analysis showed that Tibetans ( OR =2.29), breastfeeding ( OR =1.51), body dynamics outdoor daily exercise duration 30-90 min (30-<60 min ;OR = 2.03 ; 60-90 min: OR =2.22) were positively correlated with physical health of preschool children ( P <0.05). Conclusions The total physical qualification rate of preschool children aged 3-6 years old in Tibet is lower than the national average. Ethnic group, feeding pattern during infancy, and physical activity are all factors that affected the physical fitness of preschool children in Tibet. It is of great significance to improve the physique of preschool children in Tibet to promote their sustainable and healthy development.

Objective:To examine the physical health status and factors affecting individuals aged 60-69 years in certain regions of Tibet.Methods:The study analyzed data collected from 824 individuals aged 60-69 from the Xizang national physique monitoring.Chi-square test and binary logistic regression were used to identify factors that affect the physical health of this age group.Results:The study evaluated the physical fitness of elderly individuals aged 60 to 69 years old in three regions of Tibet.The results indicated that only 11 cases(1.3%)of participants had excellent physical fitness, while 47 cases(5.7%) had good physical fitness, 394 cases(47.8%)were qualified, and 372 cases(45.1%)were unqualified.The study also found significant differences in physical fitness based on age( χ2=8.367, P=0.039), sex( χ2=22.168, P<0.001), urban/rural location( χ2=12.282, P=0.006), region( χ2=60.358, P<0.001), and body mass index(BMI)( χ2=105.134, P<0.001). Specifically, individuals aged 60-64 years old had a higher proportion of qualified physique than those aged 65-69 years old.Women had better physical fitness than men, and individuals living in urban areas had better physical fitness than those in rural areas.Moreover, participants from Lhasa and Nagqu regions had better physical fitness than those from Nyingchi.Obese elderly individuals had worse physical fitness than those with normal weight(all P<0.05 for all). The study found that female gender( OR=1.765, 95% CI: 1.284-2.425), being a Lhasa( OR=2.799, 95% CI: 1.677-4.672)or Naqu resident( OR=2.730, 95% CI: 1.677-4.445), and engaging in regular exercise( OR=1.845, 95% CI: 1.078-3.160)were protective factors for physical health among individuals aged 60-69 years old.However, not working( OR=0.503, 95% CI: 0.289~0.876)and obesity( OR=0.108, 95% CI: 0.066-0.177)were identified as risk factors for physical health in this age group.Additionally, the analysis revealed significant differences in fitness ratios based on exercise frequency( χ2=18.760, P<0.001)and exercise time( χ2=15.485, P<0.001). Specifically, the fitness ratio of individuals who exercised 1-2 times and 3-4 times a week was significantly higher than that of non-exercisers.The fitness ratio of elderly individuals who exercised for 30 to 60 minutes per session was significantly higher than those who exercised for less than 30 minutes.Furthermore, those who exercised 3-4 times per week had a fitness ratio 2.58 times higher than those who did not exercise at all( P<0.05). Conclusions:This study conducted in Nyingchi, Lhasa, and Nagqu of Tibet found that 45.1% of individuals aged 60-69 years had an unqualified physique.There was a negative correlation between non-employment and BMI≥28 kg/m 2 with physical health, whereas female gender, residing in Lhasa or Nagqu, and engaging in regular physical exercise showed a positive correlation.The study suggests that physical exercise for 30-60 minutes, 3-4 times a week, can significantly improve the proportion of qualified physical fitness among the elderly.

Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into induced pluripotent stem cells (iPSCs). These studies indicated that the development of biological rhythms in ESCs might be closely associated with the maintenance and differentiation of ESCs. The core circadian gene Clock is essential for regulation of biological rhythms. Its role in the development of biological rhythms of ESCs is totally unknown. Here, we used CRISPR/CAS9-mediated genetic editing techniques, to completely knock out the Clock expression in mouse ESCs. By AP, teratoma formation, quantitative real-time PCR and Immunofluorescent staining, we did not find any difference between Clock knockout mESCs and wild type mESCs in morphology and pluripotent capability under the pluripotent state. In brief, these data indicated Clock did not influence the maintaining of pluripotent state. However, they exhibited decreased proliferation and increased apoptosis. Furthermore, the biological rhythms failed to develop in Clock knockout mESCs after spontaneous differentiation, which indicated that there was no compensational factor in most peripheral tissues as described in mice models before (DeBruyne et al., 2007b). After spontaneous differentiation, loss of CLOCK protein due to Clock gene silencing induced spontaneous differentiation of mESCs, indicating an exit from the pluripotent state, or its differentiating ability. Our findings indicate that the core circadian gene Clock may be essential during normal mESCs differentiation by regulating mESCs proliferation, apoptosis and activity.
Animals ; Apoptosis ; Base Sequence ; CLOCK Proteins ; genetics ; metabolism ; CRISPR-Cas Systems ; Cell Differentiation ; Cell Proliferation ; Cellular Reprogramming ; Circadian Clocks ; genetics ; Gene Editing ; Gene Expression Regulation ; Gene Knockout Techniques ; Hepatocyte Nuclear Factor 3-beta ; genetics ; metabolism ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Mice ; Mouse Embryonic Stem Cells ; cytology ; metabolism ; SOXB1 Transcription Factors ; genetics ; metabolism

Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies.
Amino Acid Sequence ; Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Differentiation ; genetics ; Gene Expression Regulation, Developmental ; Mesoderm ; embryology ; metabolism ; Mice, Knockout ; Molecular Sequence Data ; Pluripotent Stem Cells ; metabolism ; Sequence Homology, Amino Acid

Dysregulation of circadian rhythms associates with cardiovascular disorders. It is known that deletion of the core circadian gene Bmal1 in mice causes dilated cardiomyopathy. However, the biological rhythm regulation system in mouse is very different from that of humans. Whether BMAL1 plays a role in regulating human heart function remains unclear. Here we generated a BMAL1 knockout human embryonic stem cell (hESC) model and further derived human BMAL1 deficient cardiomyocytes. We show that BMAL1 deficient hESC-derived cardiomyocytes exhibited typical phenotypes of dilated cardiomyopathy including attenuated contractility, calcium dysregulation, and disorganized myofilaments. In addition, mitochondrial fission and mitophagy were suppressed in BMAL1 deficient hESC-cardiomyocytes, which resulted in significantly attenuated mitochondrial oxidative phosphorylation and compromised cardiomyocyte function. We also found that BMAL1 binds to the E-box element in the promoter region of BNIP3 gene and specifically controls BNIP3 protein expression. BMAL1 knockout directly reduced BNIP3 protein level, causing compromised mitophagy and mitochondria dysfunction and thereby leading to compromised cardiomyocyte function. Our data indicated that the core circadian gene BMAL1 is critical for normal mitochondria activities and cardiac function. Circadian rhythm disruption may directly link to compromised heart function and dilated cardiomyopathy in humans.

Cardiomyopathy, Dilated/metabolism* ; Humans ; Microfilament Proteins/metabolism* ; Models, Cardiovascular ; Mutation ; Myocytes, Cardiac ; Pluripotent Stem Cells/metabolism* ; Transcription Factors/metabolism* ; Troponin T/metabolism*