ObjectiveTo investigate the mechanisms by which Bushen Tongluo Jiangzhuo prescription improves renal fibrosis in rats with chronic kidney disease （CKD） through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsSeventy specific pathogen-free （SPF） Sprague-Dawley （SD） rats were randomly divided into a control group （n=15） and a modeling group （n=55）. Rats in the modeling group were administered a 2.5% adenine suspension at a dose of 200 mg·kg^-1·d^-1 by gavage for 4 weeks to establish a CKD model. Successfully modeled rats were randomly divided into a model group， an irbesartan group （20.25 mg·kg^-1·d^-1）， and Bushen Tongluo Jiangzhuo prescription low-， medium-， and high-dose groups （5.82， 11.64， and 23.28 g·kg^-1·d^-1， respectively）， with 10 rats in each group. Each group was administered an equal volume of physiological saline， the corresponding concentration of irbesartan， or Bushen Tongluo Jiangzhuo prescription by gavage for 12 weeks. Body weight and renal function indices were dynamically monitored. Serum creatinine （SCr）， blood urea nitrogen （BUN）， urine albumin-to-creatinine ratio （ACR）， 24-hour urinary total protein （24 hUTP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） levels were measured using an automatic biochemical analyzer. Renal histopathological changes were observed by hematoxylin-eosin （HE） and Masson staining. Immunohistochemistry （IHC） was used to detect the expression of PI3K， Akt， phosphorylated Akt （p-Akt）， and mTOR in renal tissues. Western blot was performed to assess the protein expression of PI3K， p-Akt， Akt， phosphorylated mTOR （p-mTOR）， and mTOR in renal tissues. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to determine the mRNA expression levels of PI3K， Akt， and mTOR in renal tissues. ResultsCompared with the model group， rats in the irbesartan group and the low-， medium-， and high-dose Bushen Tongluo Jiangzhuo prescription groups showed significantly decreased levels of SCr， BUN， ACR， 24 hUTP， IL-1β， IL-6， and TNF-α （P<0.01）. AST levels were significantly increased （P<0.01）， while no significant difference was observed in ALT levels. Histopathological examination revealed that， compared with the model group， renal tubular epithelial cell edema and necrosis and Bowman's capsule dilation were alleviated， inflammatory cell infiltration was reduced， and interstitial and glomerular fibrosis was markedly improved in all treatment groups， with the most pronounced effect observed in the high-dose Bushen Tongluo Jiangzhuo prescription group. Real-time PCR results showed that mRNA expression levels of PI3K， Akt， and mTOR were significantly downregulated in the high-dose group （P<0.01）. IHC results demonstrated that PI3K and p-Akt expression levels in renal tissues were significantly decreased in the high-dose group （P<0.01）. Western blot analysis further confirmed that the expression levels of PI3K， p-Akt/Akt， and p-mTOR/mTOR were significantly reduced in the high-dose group （P<0.01）. ConclusionBushen Tongluo Jiangzhuo prescription improves renal function indices in CKD rats， reduces collagen deposition in renal tissues， and decreases serum inflammatory factor levels. Its protective effect on renal function may be achieved by activating autophagy through downregulation of the PI3K/Akt/mTOR signaling pathway， thereby alleviating renal fibrosis.

Objective: To observe the effect of Xipayimaizibizi oral liquid (XP) in an overactive bladder (OAB) experimental rat model and to explore its pharmacological mechanisms. Methods: Network pharmacology was used to explore the potential mechanisms of action of XP. The rats underwent bladder outlet obstruction surgery and were administered the corresponding drug concentrations by gavage for 4 weeks. The study observed the body weight, water intake, bladder and kidney indices (to evaluate their general status), urination behavior pattern (to observe frequency and urgency), and urodynamics (to measure bladder parameters). Hematoxylin and eosin and Masson's trichome staining were used to observe changes in the bladder structure. Enzyme-linked immunosorbent assay was used to measure the levels of nerve growth factor, brain-derived neurotrophic factor, and acetylcholine in the urine. The key targets involved in these mechanisms were validated using reverse transcription-quantitative polymerase chain reaction, immunohistochemistry, and western blot in vivo/vitro experiments. Result: Network pharmacological analysis predicted that XP may alleviate OAB by affecting the cholinergic synapse and calcium signaling pathways. XP treatment significantly reduced the bladder index, improved urine behavior and urodynamic parameters, decreased the neurotransmitters in urine, and reduced the thickness of the bladder wall and collagen ratio. These results indicate that XP can alleviate OAB symptoms and improve the bladder structure. In vivo/vitro experiments further demonstrated that XP can inhibit targets, such as muscarinic acetylcholine receptor 2, and participate in cholinergic synapses to further regulate the parasympathetic nervous system. It can also reduce the overexpression of Ca2+ caused by agonists, inhibit targets such as transient receptor potential vanilloid type 1, and participate in calcium signaling pathways to maintain Ca2+ homeostasis. Conclusion These results suggest that XP inhibited bladder overactivity by maintaining Ca2+ homeostasis and regulating the parasympathetic nervous system.

OBJECTIVES: To investigate the effect of BRD4 inhibition on migration of esophageal squamous cell carcinoma (ESCC) cells with low acetyl-CoA carboxylase 1 (ACC1) expression. METHODS: ESCC cell lines with lentivirus-mediated ACC1 knockdown or transfected with a negative control sequence (shNC) were treated with DMSO, JQ1 (a BRD4 inhibitor), co-transfection with shNC-siBRD4 or siNC with additional DMSO or C646 (an ahistone acetyltransferase inhibitor) treatment, or JQ1combined with 3-MA (an autophagy inhibitor). BRD4 mRNA expression in the cells was detected using RT-qPCR. The changes in cell proliferation, migration, autophagy, and epithelial-mesenchymal transition (EMT) were examined with CCK8 assay, Transwell migration assay, and Western blotting. RESULTS: ACC1 knockdown did not significantly affect BRD4 expression in the cells but obviously increased their sensitivity to JQ1. JQ1 treatment at 1 and 2 μmol/L significantly inhibited ESCC cell proliferation, while JQ1 at 0.2 and 2 μmol/L promoted cell migration. The cells with ACC1 knockdown and JQ1 treatment showed increased expresisons of vimentin and Slug and decreased expression of E-cadherin. BRD4 knockdown promoted migration of ESCC cells, and co-transfection with shACC1 and siBRD4 resulted in increased vimentin and Slug expressions and decreased E-cadherin expression in the cells. C646 treatment of the co-transfected cells reduced acetylation levels, decreased vimentin and Slug expressions, and increased E-cadherin expression. Treatment with JQ1 alone obviously increased LC3A/B-II levels in the cells either with or without ACC1 knockdown. In the cells with ACC1 knockdown and JQ1 treatment, additional 3-MA treatment significantly decreased the expressions of vimentin, Slug and LC3A/B-II and increased the expression of E-cadherin. CONCLUSIONS BRD4 inhibition promotes autophagy of ESCC cells via a histone acetylation-dependent mechanism, thereby enhancing EMT and ultimately increasing cell migration driven by ACC1 deficiency.
Humans ; Cell Movement ; Transcription Factors/metabolism* ; Esophageal Neoplasms/metabolism* ; Cell Line, Tumor ; Cell Cycle Proteins ; Azepines/pharmacology* ; Epithelial-Mesenchymal Transition ; Carcinoma, Squamous Cell/metabolism* ; Esophageal Squamous Cell Carcinoma ; Triazoles/pharmacology* ; Nuclear Proteins/genetics* ; Cell Proliferation ; Acetyl-CoA Carboxylase/genetics* ; Transfection ; Autophagy ; Bromodomain Containing Proteins

Objective To explore the effect and the possible mechanism of knockdown acetyl CoA carboxylase 1(ACC1)on the migration of KYSE450 cells.Methods KYSE450 cells during the logarithmic phase were randomly divided into the shNC group,shACC1 group,shNC+AEB071 group,and shACC1+AEB071 group.The KYSE450 cells in the shNC group were transfected with empty plasmid;the KYSE450 cells in the shACC1 group were transfected with lentiviral plasmid;the KYSE450 cells in the shNC+AEB071 group were transfected with empty plasmid and then added with 5 μL of 2 mmoL·L-1 protein kinase C(PKC)inhibitor AEB071(final concentration 5 μmoL·L-1);The KYSE450 cells in the shACC1+AEB071 group were transfected with lentiviral plasmid and then added with 5 μL of 2 mmoL·L-1 PKC inhibitor AEB071(final concentration 5 μmoL·L-1).The migration of KYSE450 cells was detected by Transwell assay.The morpho-logical changes of KYSE450 cells were observed under the microscope.The expression levels of ACC1,histone H3(H3),histone H3 lysine 9 acetylation(H3K9Ac),and epithelial-mesenchymal transition(EMT)markers such as β-catenin,Vimentin and Snail were measured by Western blot.Results The migration of KYSE450 cells in the shACC1 group was significantly higher than that in the shNC group(P＜0.05);there was no significant difference in the migration ability of KYSE450 cells between the shNC+AEB071 group and the shNC group(P＞0.05);the migration of KYSE450 cells in the shACC1+AEB071 group was significantly lower than that in the shACC1 group(P＜0.05).The KYSE450 cells in the shNC group revealed an elliptical epithelial-like cell morphology;the KYSE450 cells in the shACC1 group exhibited a spindle-like interstitialcell morphology;the KYSE450 cells in the shNC+AEB071 and shACC1+AEB071 groups showed an elliptical epithelial-like cell morphology.The relative expression level of ACC1 in KYSE450 cells in the shACC1 group was significantly lower than that in the shNC group(P＜0.05),while the relative expression levels of β-catenin,Vimentin and Snail as well as the ratio of H3K9Ac/H3 were significantly higher than those in the shNC group(P＜0.05);the relative expression levels of ACC1,β-catenin,Vimentin and Snail as well as the ratio of H3K9Ac/H3 showed no significant difference between the shNC+AEB071 group and the shNC group(P＞0.05);the relative expression levels of β-catenin,Vimentin and Snail as well as the ratio of H3K9Ac/H3 in the shACC1+AEB071 group were significantly lower than those in the shACC1 group(P＜0.05);there was no significant difference in the relative expression level of ACC1 in KYSE450 cells between the shACC1+AEB071 group and the shACC1 group(P＞0.05).Conclusion Knockdown of ACC1 promotes migration of KYSE450 cells and thus aggravates the tumor,which may be mediated by PKC-related signaling pathways.

Objective To investigate the efficacy of methylprednisolone in preventing postoperative pulmonary infections in locally advanced esophageal cancer patients undergoing neoadjuvant chemotherapy combined with immunotherapy.Methods A total of 89 patients with locally advanced esophageal cancer treated at the First Affiliated Hospital of Xinxiang Medical University from January 2022 to December 2023 were selected as the research subjects.All patients underwent thoracolaparoscopic radical esophagectomy for esophageal cancer.The patients were randomly divided into an observation group(n=45)and a control group(n=44)using a random number table method.In the observation group,one patient with intraoperative thoracotomy,two patients with extensive pleural adhesion,and one patient with preoperative upper respiratory tract infection were excluded.In the control group,one patient with extensive pleural adhesion and one patient with preoperative upper respiratory tract infection were excluded.As a result,a total of 83 patients were included in the study,with 41 in the observation group and 42 in the control group.Preoperatively,a neoadjuvant treatment regimen of paclitaxel(albumin-bound)+nedaplatin in combination with camrelizumab was given to patients in both groups for 2 cycles.Patients in the control group received conven-tional anti-infection treatment after surgery,while patients in the observation group were intravenously injected with methylpred-nisolone at a dose of 1 mg·kg-1 daily from the first to the third day after surgery.Postoperative inflammatory markers,incidence of postoperative pulmonary infections,incidence of anastomotic fistula,postoperative hospital stay,and total hospitalization costs were compared between the two groups.Results There were no statistically significant differences in leukocyte count,neutrophil ratio,procalcitonin(PCT),high-sensitivity C-reactive protein(hs-CRP),and interleukin-6(IL-6)levels of patients between the two groups before treatment(P＞0.05).On day 1 and 4 after treatment,patients in the observation group had significantly higher leu-kocyte count,neutrophil ratio,PCT,hs-CRP,and IL-6 levels compared to those before treatment(P＜0.05).On postoperative day 4,the leukocyte count,neutrophil ratio,PCT,hs-CRP,and IL-6 levels were significantly lower than those on day 1 postopera-tively(P＜0.05).On postoperative days 1 and 4,the leukocyte count,neutrophil ratio,PCT,hs-CRP and IL-6 levels of patients in the control group were significantly higher than those in the preoperative period(P＜0.05),and the leukocyte count,neutro-phil ratio and hs-CRP level were significantly lower on day 4 after surgery than on day 1 after surgery(P＜0.05);the differe-nces in PCT and IL-6 level of patients between day 4 after surgery and day 1 after surgery were not statistically significant(P＞0.05).On postoperative day 1,there were no statistically significant differences in leukocyte count,neutrophil ratio,PCT,hs-CRP,and IL-6 levels between patients in the observation group and the control group(P＞0.05).On postoperative day 4,the leukocyte count,neutrophil ratio,PCT,hs-CRP,and IL-6 levels of patients in the observation group were significantly lower than those in the control group(P＜0.05).The incidence of pulmonary infections in patients in the control group and the ob-servation group was 30.9％(13/42)and 12.2％(5/41),respectively;the incidence of pulmonary infections in patients in the observation group was significantly lower than that in the control group(x2=4.298,P＜0.05).The incidence of anasto-motic fistula in patients in the observation group and the control group was 9.76％(4/42)and 21.43％(9/42),respectively;there was no statistically significant difference between the two groups(x2=2.140,P＞0.05).The postoperative hospital stay was significantly longer in the control group than in the observation group(P＜0.05),and the total hospitalization costs were significantly higher in the control group than in the observation group(P＜0.05).Conclusion Methylprednisolone can effec-tively reduce the levels of inflammatory markers and the incidence of postoperative pulmonary infections in esophageal cancer patients undergoing neoadjuvant chemotherapy combined with immunotherapy before surgery.It is a highly safe treatment thera-py without increasing the incidence of anastomotic fistula.

Concurrent chemoradiotherapy (CCRT) refers to the simultaneous treatment of chemotherapy and radiotherapy, and the effect of radiotherapy is enhanced with low-dose chemotherapy, which can reduce tumor recurrence and metastasis and improve clinical prognosis of patients. At present, the main factors for the increase of radiosensitivity of concurrent chemotherapy is that concurrent chemotherapy prevents the repair of tumor cells, and chemotherapy and radiotherapy act on different cell cycles and have synergistic effects. However, even for patients with locally advanced cervical cancer (LACC) who have undergone CCRT, the 5-year survival rate is only 60%, which is still not ideal. In order to improve the efficacy, researchers have conducted a series of exploratory studies, which consist of the combination of targeted drugs and immunodrugs, and neoadjuvant regimens before CCRT, etc. Although targeted or immunologic drugs are effective treatment of LACC, in view of the lack of large-scale evidence-based medical evidence, multi-center prospective and randomized phase III clinical trials and high-level articles are needed to improve the level of evidence-based medicine. This consensus summarizes several key evidence-based medical studies published recently, especially the clinical research progress in targeted and immunological therapies, providing reference for domestic peers.

Objective To systematically evaluate the psychological feelings and real experience of ICU nurses'moral dilemma,and the aim is to provide references for the formulation of targeted psychological intervention programs.Methods Pubmed,CINAHL,Embase,Web of Science,Cochrane Library,PsycNET,CNKI,Wanfang database,VIP database,and China Biomedical Literature Database were searched by computer to retrieve qualitative studies on psychological perceptions and real experiences of moral dilemmas of ICU nurses,and the search time was from the establishment of databases to March 2023.The quality of literature was evaluated using the 2016 version of the Australian JBI Centre for Evidence-Based Health Care Quality Evaluation Criteria for qualitative studies,and the results were integrated and analyzed by the Meta integration method.Results A total of 10 pieces of literature were included,including 3 phenomenological studies and 7 descriptive studies.The results of 27 studies were summarized into 8 new categories,and 3 integrated results were obtained:①The internal and external causes of moral dilemmas,including conflicting concepts of life and death,differences of the idea of the superior death,conflict of clinical decision-making,and insufficient organizational support;②The progressive effects of moral dilemmas,including aggravating negative emotional burden and breeding professional identity bias;③ Coping strategies for moral dilemmas,including active self-debugging and seeking support from colleagues.Conclusion It is urgent to popularize the scientific concept of life and death of patients and families,deepen the concept of superior death of ICU nurses,and alleviate the conflict of nursing and patient decision-making.It is important to pay attention to the negative emotional state of ICU nurses,improve their professional identity,improve the self-adjustment ways of ICU nurses,and to provide multi-organization support for ICU nurses.

Objective:To investigate the effects of Shenghui Yizhi Decoction on learning and memory ability of SAMP8 mice; To explore its mechanism.Methods:Totally 24 male SAMP8 mice were randomly divided into model group, donepezil hydrochloride group (1 mg/kg) and Shenghui Yizhi Decoction group (15.6 g/kg), with 8 mice in each group, and eight anti rapid aging mice (SAMR1) of the same age and strain were set as the control group. The mice in each group were given corresponding medication or the same volume of distilled water for 90 days. After administration, Morris water maze was used to detect the learning and memory ability and space exploration ability of mice. Immunohistochemistry was used to detect expression of β Amyloid protein 1-42 (Aβ 1-42) in hippocampal. ELISA was used to detect the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Real time PCR was used to detect expressions of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cysteine protease-1 (Caspase-1), and IL-1β mRNA. Western Blot was used to detect expression of NLRP3, Caspase-1and IL-1β protein. Results:Compared with model group, the spatial exploration ability and memory ability of mice in Shenghui Yizhi Decoction group and donepezil hydrochloride group were improved ( P<0.05, P<0.01), the expression of Aβ 1-42 in hippocampus decreased ( P<0.05, P<0.01), the contents of IL-1β, IL-6 and TNF-α decreased ( P<0.05, P<0.01), and the mRNA and protein expressions of NLRP3, Caspase-1 and IL-1β decreased ( P<0.05, P<0.01). Conclusion:Shenghui Yizhi Decoction can effectively improve the learning and memory ability of SAMP8 mice, and its mechanism may be related to inhibiting the activation of inflammasome and neuroinflammatory response by regulating the NLRP3/ Caspase-1/ IL-1β signaling pathway.

ObjectiveTo observe the effect of Xiaoke drink on insulin resistance in ob/ob mice and explore the mechanism. MethodEighteen ob/ob mice were randomly assigned into model， Xiaoke drink （17.68 g·kg^-1）， and atorvastatin （0.01 g·kg^-1） groups （n=6）， and six C57BL/6 mice were selected as the normal group. Mice in the normal and model groups were administrated with the same amount of distilled water. Fasting body weight， weekly food intake， and weekly water intake were measured at a fixed time. Fasting plasma glucose （FPG） and 2-hour post-load plasma glucose （2 hPG） were measured before and after 8-week intervention. After intervention， total cholesterol （TC）， triglyceride （TG）， fasting insulin （FINS）， Homeostasis Model Assessment-Insulin Resistance （HOMA-IR）， blood routine， and alkaline phosphatase （ALP） were measured. Western blot was employed to determine the expression levels of ubiquitin-specific protease 20 （USP20） and 3-hydroxy-3-methylglutaryl coenzyme A reductase （HMGCR） in the liver. The pancreas was stained with hematoxylin-eosin for observation. ResultCompared with the model group， the Xiaoke drink group showed decreased body weight of ob/ob mice （P<0.05， P<0.01）， declined growth trend of body weight （P<0.05， P<0.01）， reduced weekly average water intake， lowered levels of FPG， 2 hPG， TC， and HOMA-IR （P<0.05， P<0.01）， and down-regulated expression level of USP20 in the liver （P<0.05）. HMGCR content was positively correlated with USP20 expression. In addition， Xiaoke drink promoted the recovery of islet tissue morphology and function in ob/ob mice. ConclusionXiaoke drink can ameliorate insulin resistance in ob/ob mice by inhibiting USP20/HMGCR expression， reversing cholesterol biosynthesis process， and reducing cholesterol level.

As people's living standards improve， the development trend of diabetes has gradually become severe. Diabetes is a chronic inflammatory disease associated with abnormal expression of nuclear factor-kappa B （NF-κB） in patients. NF-κB exists in various tissue cells and participates in the regulation of a variety of genes related to immune function and inflammation. Varieties of factors can activate NF-κB when the body is stimulated by external factors， so as to produce inflammation and other reactions. Previous studies on NF-κB mainly focus on cancer， and the pathological mechanism of the treatment of diabetes by related signaling pathways and the progress of traditional Chinese medicine （TCM） treatment have not been systematically elaborated on. By referring to the relevant literature in China and abroad， it was found that NF-κB is not isolated in the development and progression of diabetes but is associated with signal molecules related to inflammation， oxidative stress， and energy metabolism， and it is involved in mediating inflammation， pancreatic β cell apoptosis， insulin signal transduction， and other physiological functions. Therefore， blocking the transmission of NF-κB signaling pathway is beneficial to the treatment of diabetes. At present， Western medicine for the treatment of diabetes mainly includes oral hypoglycemic drugs and insulin injections， but the adverse reactions are obvious. TCM has been characterized by multi-target， extensive action， and excellent curative effects in the treatment of diabetes. TCM and its compounds with functions of tonifying Qi and promoting blood circulation， regulating qi and eliminating phlegm， clearing heat and detoxifying， and nourishing Yin and moistening dryness can effectively intervene in the abnormal expression of NF-κB signaling pathway in vivo through anti-inflammatory effects. In this paper， the association between NF-κB signaling pathway and diabetes was summarized， and the modern research progress of TCM intervention of NF-κB signaling pathway in the treatment of diabetes in the past five years was reviewed， so as to lay a laboratory foundation for the study of a new pathological mechanism of diabetes based on NF-κB signaling pathway and provide new targets and research direction for the prevention and treatment of diabetes and development of related TCM.