Objective To construct HEK 293T cells that express tardigrade Dsup protein fused with green fluorescent protein copGFP in order to study the effect of Dsup protein on proliferation of HEK 293T cells.Methods The CRISPR/Cas9 gene knock-in system was constructed.The target gene fragments of Dsup,copGFP,EF1α and puromycin were amplified by PCR and inserted into pAAVS1-SFFV to construct the fusion vector of Dsup and copGFP,which was known as pAAVS1-SFFV-Dsup-copGFP-EF1α-Puro.pAAVS1-SFFV-Dsup-copGFP-EF1 α-Puro and pAAVS1-CRISPR-Cas9 vector were co-transfected into HEK 293T cells before Dsup gene was inserted into the AAVS1 region of HEK 293T cells via homologous recombination.The HEK 293T cells expressing Dsup gene were obtained following puromycin selection,flow cytometry sorting and genome identification.The expression of Dsup at mRNA and protein levels and proliferation-related genes(MCM2,MCM4,PCNA,Ki-67)were examined to investigate the effects of Dsup gene on the proliferation of HEK 293T-Dsup-copGFP cells.Results The pAAVS1-SFFV-Dsup-copGFP-EF1α-Puro recombinant vector was constructed,and the HEK 293T-Dsup-copGFP cells with Dsup gene inserted in the AAVS1 region were obtained,where both Dsup mRNA and protein were expressed.The cell proliferation rate of HEK 293T-Dsup-copGFP was higher than that of HEK 293T-Control-copGFP(P＜0.001).Further investigation revealed that the expressions of Ki-67 and MCM4 protein in HEK 293T-Dsup-copGFP were significantly higher than in the control group,indicating that the knock in of Dsup gene might enhance the proliferation ability of human cells by promoting the expression of Ki-67 and MCM4 protein.Conclusion A gene editing vector is constructed,and stable cell line HEK 293T-Dsup-copGFP for Dsup fusion expression with copGFP is established.The expression of Dsup gene in HEK 293T cells can promote cell proliferation,possibly by upregulating the expressions of Ki-67 and MCM4 protein.

Objective To investigate associations between three-dimensional(3D)morphology of cervical vertebrae and skeletal mat-uration by cone-beam computed tomography(CBCT)and establish corresponding regression models for quantitatively evaluating cervical vertebral maturation(CVM).Methods The analyzed sample consisted of 358 CBCT images(175 male,183 female),of which 277 images were randomly selected as the model development group and 81 as the performance test group.Twenty-one 3D morphological pa-rameters were defined and measured,incorporating all parts of the cervical vertebrae,including the cervical vertebral bodies,transverse processes,spinous processes,pedicles,lamina,and articular processes.The cervical vertebral maturation index(CVMI)was determined by experienced orthodontists as reference standard.Spearman's rank correlation coefficient and multivariable stepwise regression analysis were used to identify the associations and build regression models.The performance test group was employed to ex-amine each model's reliability.Paired-samples Wilcoxon signed-rank test compared the CVMI of the model prediction with the reference standard.Results Three-dimensional morphological changes in various parts of the cervical vertebrae correlated with CVMI(P＜0.05).Six 3D morphometric parameters were each recognized for male and female models,three of which were identical.The adjusted R2 was 0.899 for males and 0.902 for females,with corresponding accuracies of 85.0%and 85.4%,respectively.These models showed no difference as compared with the reference standard(P＞0.05).Con-clusion New associations were found between 3D morphology of cer-vical vertebrae and skeletal maturation.The 3D-driven morphometric CVM assessment method and corresponding regression models exhibited good credibility and high consistency with experts.

OBJECTIVE To investigate the effects of Tongxie yaofang （TXYF） on the symptoms of rats with irritable bowel syndrome with diarrhea （IBS-D） by regulating colonic tryptophan hydroxylase 1 （TPH1）， serotonin transporter （SERT） and intestinal flora. METHODS Forty-two SD rats were randomly divided into control group （7 rats） and modeling group （35 rats）. In modeling group， rat model of IBS-D was established by intragastrical administration of 0.45 g/L senna leaf solution [10 mL/（kg·d）] combined with chronic unpredictable stimulation. Thirty-five successfully modeled rats were randomly divided into model group， pinaverium bromide group [15 mg/（kg·d）] and TXYF low-dose， medium-dose and high-dose groups [3.75、7.5、15 g/（kg·d）， calculated by crude drug]， with 7 rats in each group. Each administration group was orally administered the corresponding drug， once a day， for 10 consecutive days. The general condition and weight changes of each group of rats were compared before modeling， after modeling and before administration， after the last drug intervention； the diarrhea index and visceral sensitivity were detected， and pathological changes of colon tissue were observed after modeling and before administration， after the last drug intervention. The level and expression of 5-hydroxytryptamine （5-HT）， protein and mRNA expressions of TPH1 and SERT were determined in colon tissue. The diversity and structural changes of fecal intestinal flora of rats were analyzed. RESULTS There was no significant change in colon histopathology in each group. Compared with model group， the general condition of rats in each medication group improved. The daily body weight gain of rats was significantly increased， while diarrhea index， visceral sensitivity， the expressions of 5-HT and TPH1 in colon tissue were significantly decreased； SERT expression of colon tissue was significantly increased in TXYF medium-dose and high-dose groups （P＜0.05 or P＜0.01）. The diarrhea index， colon TPH1 protein expression and colon 5-HT protein positive rate in the TXYF low-dose group decreased while the mRNA expression of SERT increased significantly （P＜0.05）. There was a dose- dependent trend in the effect of TXYF. Compared with model group， Chao1 index and Shannon index of the rats in TXYF high- dose group were significantly decreased （P＜0.05 or P＜0.01）， the beneficial bacteria such as Firmicutes and Lactobacillus increased significantly， while the pathogenic bacteria such as Proteobacteria， Escherichia-Shigella and Rikenellaceae_RC9_gut_ group decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS TXYF can decrease the level of 5-HT and improve intestinal flora disorder by inhibiting the expression of TPH1 and up-regulating the expression of SERT in colon tissue， thus promoting the symptoms of IBS-D rats.

Atherosclerosis(AS) is an important pathological feature of cardiovascular diseases such as myocardial infarction,stroke and other highly fatal diseases. Phlegm and dampness are considered to be an important pathogenesis of AS,which is difficult to heal and can cause complications. The establishment of an animal model with AS and phlegm-dampness syndrome,which could reflect the features of traditional Chinese medicine(TCM),and objective evaluation system are an important element of modern integrated TCM and western medicine research on cardiovascular diseases. It is of great significance for TCM to prevent and treat cardiovascular diseases. This article summarizes the scientific connotations of traditional Chinese and western medicine for AS and phlegm-dampness syndrome,comprehensively summarizes the current status of construction and evaluation in experimental animal model,analyzes the problems of current model,and discusses the factors of model construction and evaluation. Our aim is to establish normalized and standardized animal model with AS of phlegm-dampness syndrome.

Recently,with the rapid growth of the global population and the exhaustion of resources,exploration activities in extreme environments such as the polar regions,the outer space,the deep sea,the deep underground and highlands are becoming increasingly more frequent.This in-depth exploration of the external environment and the consequent dramatic changes in lifestyles impact on sleep,a basic life activity of humans,in ways that cannot be overlooked.the basic life activity of human beings.Sleep,a basic life activity and the result of the evolution of organisms to adapt to their environment,is closely associated with sleep homeostasis and endogenous rhythms.However,external environmental changes and lifestyle shifts in extreme environments have had a significant impact on the patterns and the quality of sleep in humans.Furthermore,this impact can lead to many physiological and psychological problems,posing a great threat to human health.In this review,we delved into the specific effects of different extreme natural environments and enclosed environments on sleep,elaborating on how these environments alter the patterns and the quality of sleep in humans.In addition,we summarized the changes in human sleep under extreme environments to help gain a better understanding of the mechanisms by which these specific environments impact human sleep.It is expected that this review will provide a solid theoretical foundation for optimizing long-term survival strategies in extreme environments and help humans adapt to and overcome the challenges posed by extreme environments more effectively.

【Objective】 To investigate the efficacy of negative pressure suction and topical testosterone cream in the treatment of simple micropenis in school-aged obese children and the effects on blood lipids and serum sex hormones. 【Methods】 A total of 79 children aged 7 to 14 (10.50±1.62) years treated and followed up during Nov.2020 and Jul.2022 were involved. The patients were randomly enrolled in the negative pressure suction group (n=39) and the topical testosterone cream group (n=40). The negative pressure suction group was treated with negative pressure suction for 30 min/time, 1 time/day, for 30 d. The topical testosterone cream group was treated with topical testosterone cream applied to the scrotum of the penis 2 times/day for 30d. The transverse and longitudinal diameter of the glans, penile flaccidity, retraction length, serum sex hormones and blood lipids were measured before and after treatment. 【Results】 In both groups, penile flaccidity, retracted length and transverse and longitudinal diameter of the glans were significantly greater at 30 days of treatment and 2 months of follow-up than those before treatment (P<0.01). The increase in penile flaccidity, retraction length and transverse and longitudinal diameter of glans at 30 days of treatment and 2 months of follow-up were greater in the topical testosterone cream group than in the negative pressure suction group (P<0.01). There were no significant differences in the blood lipids and sex hormones in the negative pressure suction group before treatment and at 30 days of treatment and 2 months of follow-up (P>0.025). In the topical testosterone cream group, at 30 days of treatment, there were significant differences in serum total cholesterol (CHOL), apolipoprotein B (Apo B), apolipoprotein A1 (Apo A1), high-density lipoprotein (HDL), low-density lipoprotein (LDL), sex hormone binding protein (SBG), estradiol (E2) and testosterone (TES) compared to those before treatment; at 2 months of follow-up, the differences in Apo A1, HDL, TES and DHS were still significant (P<0.025). 【Conclusion】 Both topical testosterone cream and negative pressure suction have significant efficacy in the treatment of micropenis in school-aged obese children, while topical testosterone cream has some side effects and retraction may occur after treatment. Negative pressure suction is safe and easy to operate, without side effects and retraction, and can be promoted for the treatment of micropenis in obese children.

OBJECTIVE: To investigate the changes in spinal-pelvic sagittal parameters from preoperative standing to prone position in old traumatic spinal fractures with kyphosis. METHODS: The clinical data of 36 patients admitted between December 2016 and June 2021 for surgical treatment of old traumatic spinal fractures with kyphosis, including 7 males and 29 females, aged from 50 to 79 years (mean, 63.9 years), were retrospectively analyzed. Lesion segments included 2 cases of T ₁₁, 12 cases of T ₁₂, 2 cases of T _{11, 12}, 4 cases of T ₁₂ and L ₁, 12 cases of L ₁, 2 cases of L ₂, 1 case of L _{2, 3}, and 1 case of L ₃. The disease duration ranged from 4 to 120 months, with an average of 19.6 months. Surgical procedures included Smith-Petersen osteotomy in 4 cases, Ponte osteotomy in 6 cases, pedicle subtraction osteotomy in 2 cases, and improved fourth level osteotomy in 18 cases; the remaining 6 cases were not osteotomized. The bone mineral density ranged from -3.0 to 0.5 T, with a mean of -1.62 T. The spinal-pelvic sagittal parameters from preoperative standing to prone positions were measured, including local kyphosis Cobb angle (LKCA), thoracic kyphosis (TK), lumbar lordosis (LL), sacral slope (SS), pelvic tilt (PT), and PI and LL mismatch (PI-LL). The kyphotic flexibility=(preoperative standing LKCA-preoperative prone LKCA)/preoperative standing LKCA×100%. Spinal-pelvic sagittal parameters were compared between standing position and prone position before operation, and Pearson correlation was used to judge the correlation between the parameters of standing position and prone position before operation. RESULTS: When the position changed from standing to prone, LKCA and TK decreased significantly ( P<0.05), while SS, LL, PT, and PI-LL had no significant difference ( P>0.05). Pearson correlation analysis showed that LL was significantly correlated with SS and PI-LL in both standing and prone positions ( P<0.05), and the correlation strength between LL and SS in prone position was higher than that in standing position. In the standing position, LKCA was significantly correlated with SS and PT ( P<0.05). However, when the position changed from standing to prone, the correlation between LKCA and SS and PT disappeared, while PT and PI-LL was positive correlation ( P<0.05). The kyphotic flexibility was 25.13%-78.79%, with an average of 33.85%. CONCLUSION For the patients of old traumatic spinal fractures with kyphosis, the preoperative LKCA and TK decrease significantly from standing position to prone position, and the correlation between spinal and pelvic parameters also changed, which should be taken into account in the formulation of preoperative surgical plan.
Male ; Female ; Humans ; Spinal Fractures/surgery* ; Standing Position ; Retrospective Studies ; Lumbar Vertebrae/surgery* ; Kyphosis/surgery* ; Lordosis/surgery*

This study designs an intermittent pneumatic pressurization device with STM32 series single chip as the core. The working state of the air pump and the plurality of air chambers is controlled by the IO port of the single chip microcomputer, and the circulating inflation of the plurality of air bags is realized. The pressure monitoring system consists of a silicon piezoresistive pressure sensor, a high-precision operational amplifier and a 12-bit AD converter which monitors the gas pressure of each gas path in real time to ensure the safety of the equipment. The system is easy to operate, simple in function, and has strong practicability.
Blood Circulation ; Cardiovascular System/physiopathology* ; Equipment Design ; Humans ; Microcomputers ; Pressure

OBJECTIVE: To establish the system software of multi-parameter monitoring by embedded Linux kernel and Qt library. METHODS: To determine the hardware system needed for the development of the system, carry out system Bootloader (Bootloader), Linux kernel, file system and Qt/Embedded (QtE) tailoring and transplantation and application development on the basis of the hardware system, and achieve the characteristic UI design. RESULTS: The changes of physiological parameters were observed in real time to improve the stability and real-time performance of the whole system and increase users' experience with QtE. CONCLUSIONS The embedded Linux+Qt multi-reference monitoring system can improve the stability, operability and functionality of real-time monitoring and multi-physiological information, and has good extensibility and maintainability.
Computers ; Electrocardiography ; Humans ; Monitoring, Physiologic/instrumentation* ; Software

Objective To observe the effects of excessive fluoride on osteocalcin and glucose metabolism in mice.Methods Thirty-two male C57 mice (body mass:18-24 g) were selected as study subjects which were randomized into four groups (8 mice in each group) according to their body mass by random number table method:0 mg/L group (control group),50 mg/L fluorine group,100 mg/L fluorine group and 150 mg/L fluorine group.Sodium fluoride in distilled water was freely taken by these animals to replicate fluorosis animal model.After 12 weeks,total osteocalcin,uncarboxylated osteocalcin,insulin and glucagon were measured by enzyme-linked immunosorbent assay,fasting blood glucose and glycated hemoglobin were measured by chemiluminescence.Results After 12 weeks of intervention with sodium fluoride,serum total osteocalcin,uncarboxylated osteocalcin,fasting blood glucose,glycated hemoglobin,insulin,and glucagon were significantly different between different groups (F =17.23,22.29,4.43,45.57,4.45,55.21,P ＜ 0.05).Total osteocalcin,uncarboxylated osteocalcin,fasting blood glucose,glycated hemoglobin,and insulin in the 100,150 mg/L fluorine groups were higher than those of control group [(30.02 ± 5.35),(35.22 ± 4.98) vs (20.23 ± 3.22) μg/L;(8.72 ± 1.34),(11.01 ± 1.02) vs (5.80 ± 1.60) μg/L;(7.53 ± 2.29),(8.53 ± 2.81) vs (4.99 ± 1.60) mmol/L;(6.74 ± 0.68),(7.12 ± 0.25) vs (4.95 ± 0.28) mmol/L;(2.65 ± 0.25),(2.74 ± 0.47) vs (2.13 ± 0.28) mU/L,P＜ 0.05].The serum glucagon levels in the 50,100,150 mg/L fluorine groups were lower than that in the control group [(20.90 ± 3.00),(23.68 ± 2.58),(21.63 ± 2.42) vs (38.61 ± 3.73) ng/L,P ＜ 0.05].Conclusion Excessive fluoride can lead to elevated osteocalcin level and abnormal glucose metabolism in mice.