Objective To explore the effect of miR-296-3p on hepatic fibrosis induced by bile duct ligation(BDL)in rats.Methods 25 SD rats were randomly divided into sham group,model(BDL)group,NC adv group,miR-296-3p adv group and miR-296-3p sponge adv group,with 5 rats in each group.The pathological changes were ob-served in rat liver tissue via HE,Masson and Sirius Red staining;the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bilirubin(TBIL)in the serum of rat in each group were detected;the expression levels of miR-296-3p,interleukin(IL)-6,IL-1 β,tumor necrosis factor(TNF)-α and smooth muscle actin(α-SMA),type Ⅰ collagen(Col1A1),and connective tissue growth factor(CTGF)mRNA in rat liver tissue were detected by qRT-PCR;the expression levels of α-SMA,Col1A1 and CTGF proteins were detected by Western blot.Immunohistochemical staining(IHC)was performed to detect the expression of α-SMA in liver tissue.Target genes of miR-296-3p was predicted by bioinformatic analysis using the online database.Zinc finger and BTB do-main-containing protein20(ZBTB20)mRNA and protein expression levels were detected.Results The pathologi-cal staining results showed that compared with sham group,a large number of infiltrated inflammatory cells and col-lagen deposition were observed in the liver tissues of rats in the BDL group and NC adv group.Compared with NC adv group,the inflammatory cells and collagen deposition decreased in the liver tissues of miR-296-3p adv group.However,in miR-296-3p sponge adv group,collagen product and inflammatory reaction increased.Compared with sham group,the contents of ALT,AST and TBIL in serum of rats in BDL group and NC adv group increased,the expression level of miR-296-3p decreased,the mRNA expression levels of IL-6,IL-1β,TNF-α increased,and the mRNA and protein expression levels of α-SMA,Col1A1 and CTGF increased(all P＜0.05).Compared with the NC adv group,the contents of ALT,AST and TBIL in serum of rats in miR-296-3p adv group decreased,the ex-pression level of miR-296-3p increased,the mRNA expression levels of IL-6,IL-1 β and TNF-α,and the mRNA and protein expression levels of α-SMA,Col1A1 and CTGF in liver tissues decreased(all P＜0.05).The results of miR-296-3p sponge adv group were opposite to those of miR-296-3p adv group(all P＜0.05).The bioinformat-ics website predicted that ZBTB20 might be a candidate target gene of miR-296-3p.Compared with sham group,the expression of ZBTB20 mRNA and protein in the liver tissues of BDL group and NC adv group increased(P＜0.05),and the expression of ZBTB20 in the liver tissues of miR-296-3p adv group decreased compared with NC adv group(P＜0.05).However,the expression of ZBTB20 in liver tissues of miR-296-3p sponge adv group in-creased(P＜0.05).Conclusion miR-296-3p expression decreases in BDL-induced hepatic fibrosis in rats,and miR-296-3p may inhibit hepatic fibrosis in BDL rats by targeting ZBTB20.

Objective To investigate the effect of miR-199a-5p on common bile duct ligation(BDL)-induced liver fibrosis in rats by regulating intestinal flora.Methods The 25 SD rats were randomly divided into five groups:the Sham group,the BDL group,the negative control adenovirus(NC adv)group,the miR-199a-5p adv group and the miR-199a-5p sponge adv group.The pathological changes of liver tissue and the degree of liver fibrosis were ob-served by HE,Masson and Sirius Red staining.The levels of aspartate aminotransferase(AST),alanine amin-otransferase(ALT),total bilirubin(TBIL)and direct bilirubin(DBIL)in serum of rats were determined by a fully automatic biochemical analyzer.The mRNA expression level of miR-199a-5p in liver tissue of rats was detec-ted by qRT-PCR.The protein expression levels of α-smooth muscle actin(α-SMA)and collagen type 1 alpha 1(COL1A1)in liver tissue of rats were detected by double immunofluorescence staining and Western blot experi-ment.Rat feces were collected for 16S rRNA high-throughput sequencing.Results The expression of miR-199a-5p was up-regulated in the liver tissue of BDL rats(P＜0.01).Compared with the NC adv group,the degree of liver injury and collagen deposition were relatively serious,the levels of AST,ALT,TBIL and DBIL in serum and the expression levels of α-SMA and COL1A1 in liver tissue increased in the miR-199a-5p adv group(all P＜0.05).However,the results of miR-199a-5p sponge adv intervention were opposite(all P＜0.05).The 16S rRNA sequencing results showed that rats treated with miR-199a-5p adv were characterized by increased diversity and richness of intestinal microbiota,changed composition of intestinal microbiota,while the results of miR-199a-5p sponge adv interfering with the bacterial community were opposite(all P＜0.05).Conclusion miR-199a-5p promotes liver fibrosis of BDL rats,and its mechanism may be related to regulating the diversity and abundance of intestinal microbiota.

Objective To investigate the effect of CXXC finger protein 4 (CXXC4) on the proliferation and apoptosis of pancreatic cancer PANC-1 cells.Methods The expression level of CXXC4 in pancreatic canc-er tissues and its relationship with prognosis and clinicopathological stage of the patients were analyzed in on-line databases.The qRT-PCR technique was used to detect the mRNA expression level of CXXC4 in human normal pancreatic ductal epithelial cell line (HPNE) and pancreatic cancer cell PANC-1,AsPC-1 and BxPC-3. si-NC,si-CXXC4,pcDNA3.1 and pCDNA3.1-CXXC4 were respectively transfected into PANC-1 cells.West-ern blot was conducted to detect the effectiveness of CXXC4 knockdown and overexpression.CCK-8,colony formation,EdU and immunofluorescence assays were conducted to analyze the effect of CXXC4 knockdown or overexpression on the proliferation and apoptosis of PANC-1 cells.The bioinformatic websites was used to predict the upstream microRNA (miRNA) of CXXC4.The Starbase database was adopted to analyze the cor-relation between miR-450b-5p and CXXC4 expression in pancreatic cancer tissues.Results The TCGA data-base results showed that the expression of CXXC4 in pancreatic cancer tissues was lowly expressed compared with in paracancerous pancreatic tissues (P<0.001),moreover which was associated with the overall survival and poor prognosis in the patients with pancreatic cancer (P<0.05).The GEPIA database analysis results showed that compared with stage Ⅰ pancreatic cancer,the CXXC4 expression in stage Ⅱ pancreatic cancer was decreased (P<0.05).Compared with HPNE cells,the CXXC4 expression in 3 kinds of pancreatic cancer cells was decreased (P<0.05).Compared with the si-NC group,the proliferation and colony formation ability of PANC-1 cells in the si-CXXC4 group were enhanced,the expressions of proliferation markers Ki67 and PC-NA were increased,and the expressions of apoptosis markers Bax,caspase-3 and caspase-9 were decreased;compared with the pcDNA3.1 group,the PANC-1 cells in the pcDNA3.1-CXXC4 group obtained the opposite results (all P<0.05).The bioinformatic websites predicted that miR-450b-5p was the upstream miRNA of CXXC4,CXXC4 in pancreatic cancer tissues was negatively correlated with the miR-450b-5p expression (r=-0.227) and miR-450b-5p in various mammalian species was highly conserved.Conclusion CXXC4 inhibits the proliferation of PANC-1 cells in pancreatic cancer and promotes theirs apoptosis.

Hip fractures are among the most common fractures in the elderly, presenting to be a leading cause of disability and mortality. Surgical treatment is currently the main treatment method for hip fractures. The incidence of perioperative malnutrition is increased after hip fractures in the elderly due to the comorbidities, decreased basal metabolic rate, accelerated protein breakdown, weakened anabolism and surgical stress. However, malnutrition not only increases the incidence of postoperative complications, but also leads to increased mortality, indicating an important role of perioperative nursing management of nutrition for the elderly patients with hip fractures. At present, there still lacks scientific guidance and application standards on perioperative nursing management of nutrition for the elderly patients with hip fractures. Therefore, the Orthopedic Nursing Committee of Chinese Nursing Association and the Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Expert consensus on perioperative nursing management of nutrition for elderly patients with hip fractures ( version 2023) according to evidence-based medical evidences and their clinical experiences. Fourteen recommendations were made from aspects of nutrition screening, nutrition assessment, nutrition diagnosis, nutrition intervention and nutrition monitoring to provide guidance for perioperative nursing management of nutrition in elderly patients with hip fractures.

[摘 要] 目的：探讨miR-203a-3p对胰腺癌BxPC-3细胞增殖、迁移和侵袭能力的影响。方法：运用癌症基因组图谱（TCGA）数据库筛选胰腺癌组织和癌旁组织中差异表达的miRNA，分析miRNA高表达与低表达时胰腺癌患者的生存率和临床分期；利用TarBase数据库分析miRNA与癌症相关的GO功能与KEGG通路，利用DIANA Tools、miRDB和TargetScan网站预测miR-203a-3p的靶基因。将miR-203a-3p mimic及NC mimic、miR-203a-3p inhibitor及NC inhibitor转染至BxPC-3细胞，用qPCR法检测胰腺癌细胞和胰腺正常上皮细胞HPNE中miR-203a-3p、miR-192-5p和miR-451a表达水平，以CCK-8法、Transwell小室法和克隆形成实验分别检测BxPC-3细胞的增殖、迁移、侵袭和集落形成能力。结果：通过TCGA数据库筛选出18个胰腺癌组织中差异表达的miRNA，其中miR-203a-3p、miR-192-5p、miR-451a具有物种保守性，且其与胰腺癌临床癌症分期、细胞周期和患者生存率相关（均P<0.05）；生物信息学网站预测显示miR-203a-3p的候选靶基因是PPM1A，PPM1A与多基因存在相互作用。miR-203a-3p、miR-192-5p和miR-451a在BxPC-3和Aspc-1细胞中均高表达（均P<0.01）。miR-203a-3p mimic组BxPC-3细胞中miR-203a-3p表达水平以及细胞增殖、迁移和侵袭能力均显著提高（均P<0.01）；miR-203a-3p inhibitor组细胞中miR-203a-3p表达水平以及细胞增殖、迁移和侵袭能力均显著降低（均P<0.01）。结论：miR-203a-3p在胰腺癌组织及细胞中均高表达，其表达与患者生存和临床分期相关，可调控BxPC-3细胞的增殖、迁移和侵袭能力。

Objective:To investigate the clinical application of noninvasive high frequency oscillatory ventilation(nHFOV)and heated humidified high flow nasal cannula(HHHFNC)in sequential ventilator evacuation of preterm infants with very low birth weight with respiratory distress syndrome(RDS).Methods:A total of 88 preterm infants of very low birth weight with RDS were enrolled in the study, who had received endotracheal intubation invasive ventilation and were ready to be replaced by non-invasive ventilation at neonatal intensive care unit(NICU)of Xuzhou Central Hospital from May 2017 to January 2020.All premature infants were routinely treated with caffeine citrate.They were divided into two groups through random number table: nHFOV/HHHFNC group(45 cases)and continuous positive airway pressure(nCPAP)/oxygen hood group(43 cases). nHFOV was given after invasive ventilator removal and HHHFNC transition was followed after nHFOV withdrawal in the nHFOV/HHHFNC group, while nCPAP was given after invasive ventilator removal and oxygen hood was followed after nCPAP withdrawal in the nCPAP/oxygen hood group.The main observation consequences were compared with arterial blood gas indexes after invasive ventilator evacuation, weaning effect and the incidence of related complications.Results:(1)There were no statistically differences between the two groups in terms of gender, gestational age, birth weight, Apgar score at 1 min and 5 min after birth, the number of glucocorticoid usage in 24 h before delivery, the number of pulmonary surfactant usage, invasive ventilation time and RDS grading( P>0.05). (2)The PaO 2, PaCO 2and oxygenation index(OI=100×MAP×FiO 2/PaO 2)of the nHFOV/HHHFNC group at 1 h and 24 h after invasive ventilator removal showed significant difference compared with the nCPAP/oxygen hood group( P<0.05). The differences as the following listed were statistically significant( P<0.05)between the two groups, including the failure rate of invasive ventilation weaning during 72 h [9%(4/45)vs.26%(11/43)], the incidence of frequent apnea [7%(3/45)vs.23%(10/43)], the failure rate of nHFOV and nCPAP noninvasive ventilation weaning [4%(2/45)vs.21%(9/43)], the oxygen-used time [12.02(9.08~12.31)d vs.14.44(11.32~13.26)d] and the incidence of nasal injury [4%(2/45)vs.26%(11/43)]. (3)The time of the first noninvasive ventilation of nHFOV and nCPAP, the incidences of lung air leakage, neonatal necrotizing enterocolitis, grade Ⅲ to Ⅳ intraventricular hemorrhage, above stageⅡretinopathy, bronchopulmonary dysplasia and the mortality rate between the two groups showed no statistical significance( P>0.05). Conclusion:nHFOV and HHHFNC used in the sequential ventilator evacuation of RDS in preterm infants with very low birth weight could improve oxygenation, reduce CO 2retention, improve the success rate of machine weaning and reduce the occurrence of apnea and nasal injury.

Objective:To explore the application effect of micro-classes combined with flipped classrooms based on short videos and images in "early rehabilitation nursing of stroke patients with hemiplegia".Methods:A total of 70 nursing students who were interns in our department from June 2019 to April 2021 were selected as the research objects, among which, 35 nursing students who entered the department from June 2019 to March 2020 were selected as the control group, using traditional teaching, and 35 nursing students who entered the department from July 2020 to April 2021 were selected as the research group, adopting the micro-classes combined with flipped classrooms teaching based on short videos and images. After the teaching, the test scores of the two groups of students, the students' satisfaction with the teaching methods, their critical thinking ability and the students' autonomous learning ability were evaluated. SPSS 17.0 was used for independent-sample t test and paired-sample t test. Results:After the teaching, the theoretical scores and clinical practice assessment scores of the research group were better than those of the control group, and the difference was statistically significant ( P<0.05); the students in the research group were more satisfied with the teaching method than those in the control group. After the teaching, the scores of 7 dimensions of critical thinking ability of the students in the research group were higher than those of the students in the control group; the scores and total scores of 4 dimensions of the self-learning ability evaluation were higher in the research group than those in the control group. Conclusion:Micro-classes combined with flipped classrooms teaching based on short videos and images can improve the teaching effect of "early rehabilitation nursing for patients with stroke and hemiplegia".

Objective:To investigate the effects of microRNA-10a(miR-10a)on the proliferation and migration of tumor-associated fibroblasts(TAFs)in the liver microenvironment, as well as on the mRNA expressions of interleukin(IL)-6, IL-8 and IL-1β in TAFs.Methods:The normal liver tissues adjacent to cancer and focal tissues of metastatic colon cancer to the liver from the same patient were collected, and then primary normal fibroblasts(NFs)and the primary cell line of TAFs were established by tissue cultivation.The NFs and TAFs were identified by morphological observation and immunofluorescence staining, and their purity was determined by flow cytometry.The real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-10a in NFs and TAFs, and then miR-10a was over-expressed in the lower ones.Subsequently, the effects of miR-10a on cell proliferation, migration and the mRNA expression levels of IL-6, IL-8 and IL-1β were detected by the cholecystokinin(CCK-8)test, wound healing assay and RT-qPCR.Results:Immunofluorescence staining showed that human cytokeratin 18(CK-18)was neither expressed in NFs nor in TAFs, while fibroblast-specific protein 1(FSP-1)was expressed in NFs and TAFs, and alpha-smooth muscle actin(α-SMA)was weakly expressed in NFs but strongly expressed in TAFs.The results of flow cytometry showed that the positive rates of α-SMA in NFs and TAFs were 95.6% and 95.3%, respectively.The mRNA expression of miR-10a in TAFs was 0.65 times of that in NFs( P<0.01). After overexpression of miR-10a, the proliferation abilities at the 3th, 4th and 5th day were lower in TAFs than in NFs( P<0.05 and 0.01), the migration abilities at 24 h and 48 h were 25% and 15% lower in TAFs than in NF group( P<0.01 and 0.05), and the mRNA levels of IL-6, IL-8, IL-1 β were 54%, 27% and 42% lower in TAFs than in NFs, respectively( P<0.01, 0.01 and 0.05). Conclusions:The overexpression of miR-10a in TAFs inhibits the cell proliferation and migration and reduces the mRNA expressions of inflammatory factors IL-6, IL-8 and IL-1β, which may be an important factor for TAFs’ inhibiting liver metastasis.

Objective: To explore the clinical value of new rapid pathological diagnosis technology in the examination of cervical lesions in primary medical institution. Methods: In the Affiliated Hospital of Yangzhou University, 25 cases of cervical scraping and 22 cases of cervical biopsy were selected.Twenty-five cases of cervical scraping scraped two wipers during operation, the results were compared with conventional pathological techniques and new rapid pathological diagnosis techniques.Twenty-two cases of cervical biopsy used a new rapid pathological diagnosis technique to perform cytological diagnosis of biopsy tissue rolls in biopsy operation.According to the results of rapid pathological diagnosis, the biopsy operation plan was adjusted to analyze the clinical value of the new rapid pathological diagnosis technique in cervical lesions in primary medical institution. Results: The new rapid pathological diagnosis technology required less space and no pollution, and the diagnosis took about 2-3 min, which did not affect the normal inspection operation.Twenty-five cases of cervical scraping were produced with new rapid pathological diagnosis technique, and the quality of cervical scraping produced with conventional pathological technique was consistent, which could satisfy the needs of pathological cytological diagnosis; twenty-two cases of cervical biopsy under the vaginal device were assisted by this technology and it could increase the positive rate of the first biopsy. Conclusion Cervical scraping and cervical biopsy are still the main methods for cervical lesions examination in primary hospitals.The new rapid pathological diagnosis technology can provide the real-time pathological diagnosis of cervical scraping, improve the positive detection rate of the first biopsy of cervical biopsy, and the cost is low, suitable for promotion in primary medical institution.

Objective: To investigate the effect of overexpression of wild-type phosphatase and tensin homolog (PTEN) deleted on chromosome 10 and its mutant G129E (exhibiting the activity of protein phosphatase and losing the activity of lipid phosphatase) on F-actin in activated hepatic stellate cells (HSCs) cultured in vitro. Methods: The activated hepatic stellate cell-T6 (HSC-T6) cells were cultured in vitro, and activated HSCs were transfected with adenovirus that carried wild-type PTEN gene and G129E gene using transient transfection. The HSCs were divided into the following groups: control group, which was transfected with DMEM medium instead of virus solution; Ad-GFP group, which was transfected with the empty adenovirus vector with the expression of green fluorescent protein (GFP); Ad-PTEN group, which was transfected with the recombinant adenovirus with wild-type PTEN gene and GFP expression; Ad-G129E group, which was transfected with the recombinant adenovirus with G129E gene and GFP expression. Western blot and quantitative real-time PCR were used to measure the protein and mRNA expression of PTEN in activated HSCs; under a laser scanning confocal microscope (LSCM), phalloidine labeled with the fluorescein tetramethylrhodamine isothiocyanate (TRITC) was used to observe the morphology of HSCs, distribution and fluorescence intensity of F-actin, and changes in pseudopodia and stress fibers, and a calcium fluorescence probe (Rhod-2/AM) was used to measure the changes in Ca²⁺ concentration in HSCs. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference test was used for comparison between two groups. Results: Wild-type PTEN and G129E genes were highly expressed in activated HSCs. In the control group and the Ad-GFP group, HSCs had a starlike or polygonal shape, F-actin was reconfigured and formed a large number of stress fibers which stretched across the whole cell, and layered pseudopodia were seen around the cell. In the Ad-PTEN group and the Ad-G129E group, the HSCs had a fusiform shape, F-actin was mainly seen around the cell, a small number of stress fibers were seen inside the cell, and layered pseudopodia around the cell disappeared. The Ad-PTEN group and the Ad-G129E group had significant reductions in the fluorescence intensity of F-actin compared with the control group and the Ad-GFP group (357.67±13.39/377.25±14.55 vs 961.87±27.33/954.68±20.71, F = 1783.486, P < 0.05), while there were no significant differences between the Ad-PTEN group and the Ad-G129E group, as well as between the control group and the Ad-GFP group (P > 0.05). The Ad-PTEN group and the Ad-G129E group had significant reductions in the relative concentration of Ca²⁺ compared with the control group and the Ad-GFP group (251.60±90.88/352.18±146.01 vs 1953.95±132.99/1937.57±115.17, F = 834.988, P < 0.05), while there were no significant differences between the Ad-PTEN group and the Ad-G129E group, as well as between the control group and the Ad-GFP group (P > 0.05). Conclusion The overexpressed wild-type PTEN and its mutant G129E can significantly inhibit the formation and reconfiguration of cytoskeletal protein F-actin and reduce the concentration of Ca2+ in activated HSCs in vitro. In addition, there are no significant differences in the above effects between wild-type PTEN and G129E.