Objective To explore the value of diffusion-weighted imaging(DWI),amide proton transfer-weighted imaging(APTWI)and glucose chemical exchange saturation transfer imaging(glucoCEST)for predicting lymph node metastasis of rectal cancer.Methods Sixty cases of rectal cancer confirmed by pathology were retrospectively enrolled and divided into metastasis group(n=36)and non-metastasis group(n=24)based on the presence or absence of lymph node metastasis.The apparent diffusion coefficient(ADC)value,as well as the values of magnetization transfer ratio asymmetry at 3.50 ppm and 1.20 ppm(MTRasym3.50 ppm and MTRasym1.20ppm)of the lesions were respectively obtained based on DWI,APTWI and glucoCEST and then compared between groups.Receiver operating characteristic curve was drawn,the area under the curve(AUC)was calculated to evaluate the efficacy of each parameter alone and their combination for predicting lymph node metastasis of rectal cancer.Results In metastasis group,ADC value of the lesions was lower,while MTRasym3.50 ppm and MTRasym1.20 ppm values were both higher than those in non-metastasis group(all P＜0.05).AUC of the combination of the above parameters for predicting lymph node metastasis of rectal cancer based on ADC,MTRasym3.50ppm and MTRasym1.20 ppm values was 0.970,significantly higher than that of each single parameter(0.858,0.804 and 0.755,respectively;Z=2.442-3.231,all P＜0.05).Conclusion DWI,APTWI and glucoCEST could all be used to predicte lymph node metastasis of rectal cancer.Combination of ADC,MTRasym3.50 ppm and MTRasym1.20 ppm had the highest predictive efficacy.

Objective To explore the value of diffusion-weighted imaging(DWI),amide proton transfer-weighted imaging(APTWI)and glucose chemical exchange saturation transfer imaging(glucoCEST)for predicting lymph node metastasis of rectal cancer.Methods Sixty cases of rectal cancer confirmed by pathology were retrospectively enrolled and divided into metastasis group(n=36)and non-metastasis group(n=24)based on the presence or absence of lymph node metastasis.The apparent diffusion coefficient(ADC)value,as well as the values of magnetization transfer ratio asymmetry at 3.50 ppm and 1.20 ppm(MTRasym3.50 ppm and MTRasym1.20ppm)of the lesions were respectively obtained based on DWI,APTWI and glucoCEST and then compared between groups.Receiver operating characteristic curve was drawn,the area under the curve(AUC)was calculated to evaluate the efficacy of each parameter alone and their combination for predicting lymph node metastasis of rectal cancer.Results In metastasis group,ADC value of the lesions was lower,while MTRasym3.50 ppm and MTRasym1.20 ppm values were both higher than those in non-metastasis group(all P＜0.05).AUC of the combination of the above parameters for predicting lymph node metastasis of rectal cancer based on ADC,MTRasym3.50ppm and MTRasym1.20 ppm values was 0.970,significantly higher than that of each single parameter(0.858,0.804 and 0.755,respectively;Z=2.442-3.231,all P＜0.05).Conclusion DWI,APTWI and glucoCEST could all be used to predicte lymph node metastasis of rectal cancer.Combination of ADC,MTRasym3.50 ppm and MTRasym1.20 ppm had the highest predictive efficacy.

Chinese hamster ovary (CHO) cells play an irreplaceable role in biopharmaceuticals because the cells can be adapted to grow in suspension cultures and are capable of producing high quality biologics exhibiting human-like post-translational modifications. However, gene expression regulation such as transgene silencing and epigenetic modifications may reduce the recombinant protein production due to the decrease of expression stability of CHO cells. This paper summarized the role of epigenetic modifications in CHO cells, including DNA methylation, histone modification and miRNA, as well as their effects on gene expression regulation.
Cricetinae ; Animals ; Humans ; Cricetulus ; CHO Cells ; Epigenesis, Genetic/genetics* ; DNA Methylation ; Gene Expression Regulation ; Recombinant Proteins/genetics*

ObjectiveTo explore the clinical efficacy and safety of Qifu Yixin prescription in treating chronic heart failure in the patients with the syndrome of heart Qi deficiency， so as to provide clinical evidence for the treatment of chronic heart failure with this prescription and promote the clinical application and transformation of this prescription. MethodA total of 106 chronic heart failure patients with the syndrome of heart Qi deficiency who met the criteria in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine （TCM） from September 2022 to May 2023 were selected and randomized into an observation group （53 cases） and a control group （53 cases）. Both groups received routine Western medicine treatment. In addition， the observation group received Qifu Yixin prescription， while the control group received placebo. The treatment course for both groups was 12 weeks. The New York heart association （NYHA） cardiac function grading， N-terminal pro-B-type natriuretic peptide （NT-proBNP）， soluble growth-stimulating expression gene 2 （sST2）， left ventricular ejection fraction （LVEF）， the ratio of early diastolic maximum mitral flow velocity （E） to early diastolic mitral annular motion velocity （e'） （E/e'）， left ventricular end diastolic diameter （LVEDD）， TCM syndrome scores， 6-minute walking test （6MWT）， Minnesota Living with Heart Failure Questionnaire （MLHFQ）， and Kansas City Cardiomyopathy Questionnaire （KCCQ） were determined before and after treatment. ResultFinally， 102 patients were included for analysis， including 51 patients in the observation group and 51 patients in the control group. After treatment， 42 patients in the observation group showed improved cardiac function grading， with a total response rate of 82.35%， and 32 patients in the control group showed improved cardiac function grading， with a total response rate of 62.75%. The total response rate regarding the cardiac function in the observation group was higher than that in the control group （χ²=4.923， P<0.05）. The observation group outperformed the control group in lowering the NT-proBNP level， elevating LVEF， decreasing the E/e' ratio （P<0.05）， reducing LVEDD and sST2 levels， and recovering TCM syndrome score， 6MWT score， MLHFQ score， and KCCQ score （P<0.05）. None of the safety indexes in the two groups showed abnormal values before and after treatment， and no serious adverse reaction was observed. ConclusionQifu Yixin prescription can improve the heart function， exercise tolerance， and quality of life and alleviate the TCM syndrome of the chronic heart failure patients with the syndrome of heart Qi deficiency by inhibiting myocardial fibrosis.

Objective:To understand the current situation of nurses' willingness to engage in telecare in county-level hospitals in Henan Province and analyze its influencing factors.Methods:Using the convenient sampling method, a total of 1 300 clinical nurses from 6 Class Ⅱ Grade A county-level hospitals in Zhengzhou, Hebi, Zhoukou and other cities in Henan Province were selected as the research objects from August 2022 to January 2023. General Data Questionnaire and Clinical Nurses' Willingness to Engage in Telecare Questionnaire were used to investigate the research objects. Non-parametric test and multiple linear regression were used to analyze the influencing factors of clinical nurses' willingness to engage in telecare.Results:A total of 1 300 questionnaires were sent out in this study, and 1 183 were effectively collected, with an effective recovery rate of 91.00% (1 183/1 300). The total score of Clinical Nurses' Willingness to Engage in Telecare Questionnaire of 1 183 nurses in county-level hospitals was [105.00 (98.00, 112.00) ] points, and all items were scored [3.50 (3.27, 3.73) ]. Multiple linear regression analysis showed that professional title, post, whether they knew about telecare, and the number of times of receiving training on nursing information each year were the influencing factors of nurses' willingness to engage in telecare in county-level hospitals ( P<0.05) . Conclusions:In Henan Province, 1 183 nurses in county-level hospitals have a high willingness to engage in telecare. Relevant departments should further strengthen the publicity of relevant contents of telecare, carry out practical operation training related to nursing information, improve the nursing information ability of nurses in county-level hospitals and strengthen the awareness of health management services, so as to promote the successful implementation of telecare services in rural areas.

Objective:To investigate the effect of dydrogesterone (DYD) on the ultrastructure of the endometrium in the window of implantation.Methods:We prospectively studied endometrium tissues of patients who underwent in vitro fertilization and embryo transfer (IVF-ET) from July 2020 to July 2022 at the Center for Reproductive Medicine of Zhejiang Provincial People's Hospital. The ultrastructure of mid-luteal endometrium was examined by scanning electron microscopy and transmission electron microscopy. Firstly, we used self-control study to compare the ultrastructure of endometrium between the natural cycle group and the DYD 40 mg/d group in 20 patients with male factor infertility. Secondly, we used self-control study to compare the ultrastructure of the endometrium between the DYD 40 mg/d group and the DYD 80 mg/d group in 43 patients with recurrent implantation failure (RIF). Results:There were no significant differences in endometrial thickness, ultrasound grading, positivity rate of nucleolar channel system (NCS), pinopode score and secretor status score, in addition to number and morphology score of mitochondria, between the natural cycle group and the DYD 40 mg/d group (all P>0.05). In RIF patients, the DYD 80 mg/d group rendered significantly higher pinopode score (-46.26±19.97), NCS positivity rate [65.12% (28/43)] and mitochondrial morphology score [0 (0, 1)] than those in the DYD 40 mg/d group [-67.62±15.94, P<0.001; 39.53% (17/43), P=0.043; 0 (0, 0), P=0.023], while other characteristics were not significantly different between the two groups (all P>0.05). Conclusion:The ultrastructure of endometrium in DYD supplementation cycle was similar to that in the natural cycle. Increasing the dosage of DYD significantly improved pinopode maturity, NCS positivity rate and mitochondrial morphology of the endometrium in RIF patients.

Objective:To investigate the effect of dydrogesterone (DYD) on the ultrastructure of the endometrium in the window of implantation.Methods:We prospectively studied endometrium tissues of patients who underwent in vitro fertilization and embryo transfer (IVF-ET) from July 2020 to July 2022 at the Center for Reproductive Medicine of Zhejiang Provincial People's Hospital. The ultrastructure of mid-luteal endometrium was examined by scanning electron microscopy and transmission electron microscopy. Firstly, we used self-control study to compare the ultrastructure of endometrium between the natural cycle group and the DYD 40 mg/d group in 20 patients with male factor infertility. Secondly, we used self-control study to compare the ultrastructure of the endometrium between the DYD 40 mg/d group and the DYD 80 mg/d group in 43 patients with recurrent implantation failure (RIF). Results:There were no significant differences in endometrial thickness, ultrasound grading, positivity rate of nucleolar channel system (NCS), pinopode score and secretor status score, in addition to number and morphology score of mitochondria, between the natural cycle group and the DYD 40 mg/d group (all P>0.05). In RIF patients, the DYD 80 mg/d group rendered significantly higher pinopode score (-46.26±19.97), NCS positivity rate [65.12% (28/43)] and mitochondrial morphology score [0 (0, 1)] than those in the DYD 40 mg/d group [-67.62±15.94, P<0.001; 39.53% (17/43), P=0.043; 0 (0, 0), P=0.023], while other characteristics were not significantly different between the two groups (all P>0.05). Conclusion:The ultrastructure of endometrium in DYD supplementation cycle was similar to that in the natural cycle. Increasing the dosage of DYD significantly improved pinopode maturity, NCS positivity rate and mitochondrial morphology of the endometrium in RIF patients.

ObjectiveTo observe the behavioral and pain threshold alterations， as well as the changes in indexes related to depression and pain in the serum and central system in mice stressed by maternal separation and chronic neuropathic pain， and explore the underlying mechanism of Wenyang prescription （WY）， Jieyu prescription （JY）， and Wenyang Jieyu prescription （WYJY） in improving depression and pain sensitivity. MethodThe birth date of mice was recorded as PD0. After birth， the mice were divided into a blank group and an experimental group. The neonatal mice in the experimental group underwent maternal separation in PD5-14 at 8 h·d^-1. After ablactation， the mice were divided into a maternal separation group， a WY group （Erxian decoction， 5.84 g·kg^-1）， a JY group （Xiaoyaosan， 12.00 g·kg^-1）， a WYJY group （16.68 g·kg^-1）， and a fluoxetine group （2.60 mg·kg^-1）， with 15 mice in each group. Meanwhile， 15 male mice of the same age without maternal separation were assigned to the normal control group. Mice in the blank group and the maternal separation group were fed on a regular chow diet in PD21-PD90， while the remaining groups were fed on the corresponding drugs. In PD91， sciatic nerve ligation was performed to induce a model of maternal separation and chronic neuropathic pain. The open field test was used to observe the depression-like behaviors of mice in each group， and the mechanical and temperature pain thresholds were measured to detect the pain sensitivity of mice in each group. The serum levels of corticosterone （CORT）， substance P， and β-endorphin （β-EP） were determined by enzyme-linked immunosorbent assay （ELISA）， and the expression of the glucocorticoid receptor （GR） in the amygdala and β-EP protein in the hypothalamus was detected by immunohistochemistry. The mRNA expression levels of amygdala GR gene （Nr3c1）， FK506 binding protein 5 gene （FKBP5）， metabolic glutamate receptor 5 gene （GRM5）， and brain-derived neurotrophic factor （BDNF） were detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultCompared with the blank group， the maternal separation group showed reduced stay time and total distance traveled in the 5-min open field test （P<0.01）， reduced mechanical pain threshold （P<0.01）， increased serum CORT and β-EP （P<0.01）， declining FKBP5 mRNA expression （P<0.01）， and increased hypothalamic β-EP expression （P<0.05）. Compared with the maternal separation group， the groups with drug intervention showed prolonged stay time （P<0.05， P<0.01） and up-regulated pain thresholds to different degrees. The total distance traveled in the 5-min open field test increased in the WY group， the WYJY group， and the fluoxetine group （P<0.05， P<0.01）. The JY group showed decreased serum CORT （P<0.01）， reduced β-EP ， and increased BDNF mRNA （P<0.01）. Nr3c1 and GRM5 mRNA decreased in the WY group （P<0.05， P<0.01）. The WYJY group showed decreased serum CORT （P<0.05）and decreased Nr3c1， GRM5， and BDNF mRNA （P<0.05， P<0.01）. The levels of β-EP expression were elevated to different degrees in the groups with drug intervention， but the differences were not significant. The levels of GR expression in the WY group， the JY group， and the WYJY group increased （P<0.05）. ConclusionWYJY can inhibit central pain sensitization and regulate hypothalamic-pituitary-adrenal gland （HPA） axis function by enhancing the expression of GR in the amygdala and inhibiting neuroplasticity and excitability in the amygdala to relieve depression-like behaviors and improve somatic hyperalgesia.

Teaching rounds is an important part of clinical teaching practice, therefore, we established a demonstration team for teaching ward rounds. By formulating standard operation procedure for teaching rounds and encouraging innovation on teaching models, the team played a demonstration role in the clinical teaching rounds, which not only made up the shortcomings in teaching, but also improved the teaching level of clinical teachers and the quality of clinical training.

The aim of this study is to investigate the effect of the chimeric intron in different directions on the expression of the nerve growth factor (NGF) in recombinant Chinese hamster ovary (CHO) cells. The chimeric intron that contained the splice sequence of the first intron of the human β-globin and the human immunoglobulin heavy chain variable region intron was used. NGF gene was cloned into the expression vectors containing the chimeric intron in the forward or reverse direction, followed by transfecting into CHO cells, and screened under G418 to produce the stable transfected CHO cells. Fluorescence quantitative PCR, ELISA, and Western blotting were performed to detect the recombinant NGF gene expression in CHO cells. The results showed that the chimeric introns could significantly enhance the expression of NGF in recombinant CHO cells. Moreover, the enhancing effect on NGF expression level by the intron in the forward direction showed stronger than that of the reverse direction both at mRNA and protein level. In conclusion, the chimeric intron could increase NGF expression in stably transfected CHO cells and the effect is associated with the direction of the intron insertion.
Animals ; Animals, Genetically Modified ; CHO Cells ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Introns ; Transfection