OBJECTIVE To compare the antihypertensive efficacy of sacubitril/valsartan versus benazepril in patients with perimenopausal hypertension， as well as their impacts on ventricular remodeling and inflammatory fibrosis. METHODS A total of 206 perimenopausal hypertensive patients in our hospital from January 1， 2023 to December 30， 2024 were retrospectively included.These patients were enrolled and divided into benazepril group （105 cases） and sacubitril/valsartan group （101 cases）. Benazepril group received Benazepril hydrochloride tablet， and sacubitril/valsartan group received Sacubitril valsartan sodium tablet. All patients were treated for 6 months. The blood pressure（systolic blood pressure and diastolic blood pressure） and blood pressure control status before and after treatment， echocardiographic indicators （left ventricular ejection fraction， left ventricular mass index， relative wall thickness， and early-diastolic peak transmitral flow velocity/early-diastolic peak velocity of the mitral annulus）， inflammatory fibrosis related indicators（high-sensitivity C-reactive protein，ratio of monocytes to lymphocytes，and ratio of neutrophils to lymphocytes）， as well as the occurrence of adverse reactions（hypotension，hyperkalemia，and angioedema） were observed in both groups before and after treatment. RESULTS The blood pressure control rate was significantly higher in the sacubitril/valsartan group than in benazepril group （ P ＜0.05）. After treatment， the blood pressure， echocardiographic indicators（except for left ventricular ejection fraction） ，and inflammatory fibrosis related indicators were significantly lower than those before treatment within the same group， and the sacubitril/valsartan group were significantly lower than the benazepril group （ P ＜0.05）. There were no statistically significant differences in the incidence of hypotension， hyperkalemia， angioedema， and overall adverse drug reactions between the two groups （ P ＞0.05）. CONCLUSIONS Compared with benazepril， sacubitril/valsartan provides superior blood-pressure control， reverses ventricular remodeling， attenuates inflammatory fibrosis in perimenopausal hypertensive patients， while maintaining a similar safety profile.

Objective : To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect. Methods : Brain tissue from 4-week-old Sprague Dawley rats was screened, pre-digested and solidified to obtain brain microvascular segments. These segments were subsequently placed in a CO2incubator for primary culture. The target cells were identified by cell morphology and immunocytochemical staining for factor Ⅷ-related antigen. Results : After a 48-hour culture periodin vitro, the short spindle cells crawled out from around the brain microvascular segments. After 72 hours, island-like cell culsters formed. After 96 hours the clusters fused and the cells formed a typical monolayer, cobble stone-like, and mosaic arrangement. Factor Ⅷ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red, indicating positive expression; DAB stained the nucleus, showing blue-dark. Conclusion The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.

ObjectiveThis study aims to enhance of the farnesyl pyrophosphate（FPP） pool in Saccharomyces cerevisiae by heterologously expressing different farnesyl diphosphate synthases（FPSs） from various plants， thereby increasing the production of terpenoid compounds by the engineered yeast. MethodsRNA from mixed samples of roots， stems， and leaves of seven plants including Arabidopsis thaliana， Rosa rugosa， Artemisia annua， Centella asiatica， Humulus lupulus， Medicago sativa， and Panax ginseng was extracted by column chromatography and reverse transcribed into the first strand of complementary DNA（cDNA）， and based on the transcriptome data of the seven species of plants， sequence-specific primers were designed for CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS， the full-length of the genes was cloned， and the genes were analyzed for bioinformatics in order to construct a pESC yeast shuttle vector. These seven plant-derived FPSs were further heterologously expressed in the previous constructed β-elemene-producing yeast， and the yield of β-elemene was indicated for their catalytic acivities. ResultsThe coding sequences of CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS were all of 1 021 bp in length and encoding 301 amino acids， all of which were similarly related to the endogenous FPS-encoding gene（ERG20） in S. cerevisiae. After heterologous expression， RrFPS was identified as the most effective in catalyzing the synthesis of FPP from isopentenyl pyrophosphate（IPP） and dimethylallyl pyrophosphate（DMAPP）. Compared to the control strains， the RrFPS overexpressed yeast strains YB-1-Rr and YB-3-Rr increased the production of β-elemene by 231.25% and 189.3%， respectively. ConclusionBy comparing the functions of FPS-encoding genes from seven different plant sources， it is determined that the protein encoded by the RrFPS from R. rugosa has the best catalytic ability， which can provide key genetic elements for the construction of engineered yeast strain constructs with high terpenoid production.

ObjectiveTo investigate the mechanism of Huazhuo Jiedu prescription in treating ulcerative colitis （UC） by regulating mitophagy. MethodsThe genes related to mitophagy and UC were retrieved from GeneCards， and then the common genes of mitophagy and UC were analyzed by metascape to identify the genes related to mitophagy in UC. Animal experiments were carried out to decipher the mechanism by which Huazhuo Jiedu prescription treated UC by regulating mitophagy. Sixty C57BL/6 male mice were randomized into normal， model， high-， medium-， and low-dose （50， 25， 12.5 g·kg^-1， respectively） Huazhuo Jiedu prescription， and mesalazine （0.52 g·kg^-1·d^-1） groups， with 10 mice in each group. After successful modeling by the dextran sulfate sodium-free drinking method， the colonic mucosal damage was observed by hematoxylin-eosin staining， and the ultracellular structure of colon mucosa was observed by transmission electron microscopy. The expression levels of mitophagy-related proteins PTEN-induced putative kinase 1 （PINK1） and Parkin protein were determined by Western blot. The expression of prohibitin 2 （PHB2）， ubiquitin-specific protease 15 （USP15）， ubiquitin-specific protease 30 （USP30） in the colon tissue was detected by immunofluorescence （IF）. ResultsAll the drug intervention groups showed ameliorated pathological manifestations of the colonic mucosa and improved mitochondrial structures in UC mice. Compared with the normal group， the model group demonstrated up-regulated protein levels of PINK1 and Parkin （P<0.05）， enhanced average fluorescence intensity of PHB2 （P<0.05）， and weakened average fluorescence intensity of USP15 and USP30 （P<0.05）. Compared with the model group， the mesalazine group and the high- and medium-dose Huazhuo Jiedu prescription groups showcased down-regulated protein levels of PINK1 and Parkin （P<0.05）， decreased average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. The low-dose Huazhuo Jiedu prescription group showed down-regulated protein levels of PINK1 and Parkin （P<0.05）， weakened average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. ConclusionHuazhuo Jiedu prescription can attenuate the intestinal mucosal injury and improve the mitochondrial cell ultrastructure in UC mice by regulating the expression of PINK1-Parkin pathway and inhibiting excessive mitophagy.

ObjectiveTo investigate the mechanism of Huazhuo Jiedu prescription in treating ulcerative colitis （UC） by regulating mitophagy. MethodsThe genes related to mitophagy and UC were retrieved from GeneCards， and then the common genes of mitophagy and UC were analyzed by metascape to identify the genes related to mitophagy in UC. Animal experiments were carried out to decipher the mechanism by which Huazhuo Jiedu prescription treated UC by regulating mitophagy. Sixty C57BL/6 male mice were randomized into normal， model， high-， medium-， and low-dose （50， 25， 12.5 g·kg^-1， respectively） Huazhuo Jiedu prescription， and mesalazine （0.52 g·kg^-1·d^-1） groups， with 10 mice in each group. After successful modeling by the dextran sulfate sodium-free drinking method， the colonic mucosal damage was observed by hematoxylin-eosin staining， and the ultracellular structure of colon mucosa was observed by transmission electron microscopy. The expression levels of mitophagy-related proteins PTEN-induced putative kinase 1 （PINK1） and Parkin protein were determined by Western blot. The expression of prohibitin 2 （PHB2）， ubiquitin-specific protease 15 （USP15）， ubiquitin-specific protease 30 （USP30） in the colon tissue was detected by immunofluorescence （IF）. ResultsAll the drug intervention groups showed ameliorated pathological manifestations of the colonic mucosa and improved mitochondrial structures in UC mice. Compared with the normal group， the model group demonstrated up-regulated protein levels of PINK1 and Parkin （P<0.05）， enhanced average fluorescence intensity of PHB2 （P<0.05）， and weakened average fluorescence intensity of USP15 and USP30 （P<0.05）. Compared with the model group， the mesalazine group and the high- and medium-dose Huazhuo Jiedu prescription groups showcased down-regulated protein levels of PINK1 and Parkin （P<0.05）， decreased average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. The low-dose Huazhuo Jiedu prescription group showed down-regulated protein levels of PINK1 and Parkin （P<0.05）， weakened average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. ConclusionHuazhuo Jiedu prescription can attenuate the intestinal mucosal injury and improve the mitochondrial cell ultrastructure in UC mice by regulating the expression of PINK1-Parkin pathway and inhibiting excessive mitophagy.

Objective To analyze the influencing factors of poor incision healing in postoperative patients with breast cancer.Methods The clinical data of 150 patients with breast cancer diagnosed by the Department of Nail Milk surgery of the Fifth affiliated Hospital of Xinjiang Medical University from January 2016 to December 2023 were retrospectively analyzed.According to the postoperative wound healing,the patients were divided into two groups:good healing group(n=94)and poor healing group(n=56).The general data,operation-related data and clinicopathological characteristics of the two groups were collected for univariate analysis,and the single-factor indexes with statistical significance were analyzed by multivariate Logistic analysis to screen the risk factors of poor incision healing after operation.The statistically significant indexes of regression analysis were analyzed by receiver operating characteristic(ROC)curve to further explore its predictive value in poor incision healing after breast cancer operation.Results The data of the two groups were compared and analyzed.Multivariate Logistic regression analysis showed that BMI,diabetes,age,axillary lymph node dissection and hemoglobin were independent risk factors for poor incision healing(P＜0.05).ROC curve analysis of independent risk factors showed that when the age was more than 57.5,the area under the curve(AUC)was 0.635,the sensitivity was 55.4％,the specificity was 68.1％,and the critical value was 57.5.When BMI＞24.9 kg/m2,the area under the curve(AUC)was 0.735,the sensitivity was 87.5％,the specificity was 61.7％,and the critical value was 24.9(P＜0.001).When hemoglobin＜101.5 g/L,the area under the curve(AUC)was 0.829,the sensitivity was 57.1％,the specificity was 94.7％,and the critical value was 101.5 g/L(P＜0.001).Conclusion BMI,diabetes,hemoglobin,age and axillary lymph node dissection are independent risk factors for poor wound healing after breast cancer operation.When BMI＞24.9 kg/m2,age＞57.5 years old and hemoglobin＜101.5 g/L,it can predict the occurrence of poor incision healing in patients with breast cancer.

Objective:To investigate the risk factors of hypoparathyroidism (HPT) and hypocalcemia in patients undergoing total thyroidectomy (TT), and to explore the changes of parathyroid hormone and blood calcium after TT.Methods:The clinical data of 101 patients undergoing TT from November 2018 to September 2022 in the First Hospital of Hebei Medical University were retrospectively analyzed. The basic clinical data were recorded. The blood calcium and parathyroid hormone levels were measured before surgery and 1 d, 1 week after surgery. The occurrence of postoperative hypocalcemia was recorded. According to postoperative parathyroid hormone level, the patients were divided into control group (normal parathyroid function) and HPT group (reduced parathyroid hormone level). The patients with postoperative hypocalcemia symptoms were classified as the hypocalcemia symptoms group, and the patients without postoperative hypocalcemia symptoms were classified as the non-hypocalcemia symptoms group. Multivariate Logistic regression was used to analyze the independent risk factors of HPT and hypocalcemia in TT patients.Results:The postoperative parathyroid hormone level decreased in 41 cases (HPT group) and normal in 60 cases (control group). There were 24 patients with postoperative hypocalcemia symptoms (hypocalcemia symptoms group) and 77 patients without postoperative hypocalcemia symptoms (non-hypocalcemia symptoms group). The rate of using bipolar electric coagulation forceps in HPT group was significantly lower than that in control group: 31.71% (13/41) vs. 76.67% (46/60), while the rate of central lymph node dissection was significantly higher than that in control group: 82.93% (34/41) vs. 60.00% (36/60), and there were statistical differences ( P<0.01 and <0.05). Multivariate Logistic regression analysis result showed that TT combined with unilateral or bilateral central lymph node dissection was an independent risk factor for HPT in TT patients ( OR = 1.706 and 1.501, 95% CI 1.019 to 2.856 and 1.052 to 2.140, P<0.05). The preoperative serum calcium, postoperative serum calcium and postoperative parathyroid hormone in hypocalcemia symptoms group were significantly lower than those in hypocalcemia symptoms group: (2.32 ± 0.11) mmol/L vs. (2.37 ± 0.11) mmol/L, (2.16 ± 0.21) mmol/L vs. (2.25 ± 0.18) mmol/L and 3.00 (1.00, 5.45) ng/L vs. 19.90 (8.50, 33.80) ng/L, and there were statistical differences ( P<0.05 or <0.01). Multivariate Logistic regression analysis result showed that postoperative parathyroid hormone was an independent risk factor of hypocalcemia symptoms in TT patients ( OR = 0.927, 95% CI 0.883 to 0.974, P<0.01). In patients with HPT, the blood calcium at 1 week after surgery was significantly lower than that at 1 d after surgery: (2.07 ± 0.19) mmol/L vs. (2.17 ± 0.25) mmol/L, and there was statistical difference ( t = 2.05, P<0.05); the parathyroid hormone at 1 week after surgery was significantly higher than that at 1 d after surgery: 8.30 (3.55, 19.55) ng/L vs. 3.60 (1.00, 6.85) ng/L, and there was statistical difference ( Z = - 3.78, P<0.01). Conclusions:When performing TT, standardizing the surgical techniques, reducing unnecessary central lymph node dissection, and using bipolar electric coagulation forceps as much as possible can help to reduce the occurrence of postoperative HPT. The levels of postoperative parathyroid hormone and blood calcium should be promptly detected, the change of both should be given attention, and do a good job in preventing and treating hypocalcemia.

Objective:To establish a simple and efficient method for in vitro extraction,culture and identification of brain microvascular endothelial cells(BMECs)of rabbits.Method:The cerebral cortex of 3-week-old rabbits was cut up,sieved and digested with type II collagenase to obtain pure brain microvascular segments for primary culture and identification.Results:The cultured target cells grew in a typical"cobblestone-like"arrangement.Immunohistochemi-cal staining for von Willebrand factor(vWF)related antigen showed that the cytoplasm was presented as brownish red and the expression was positive.Conclusion:The physical sieving method combined with the chemical enzyme diges-tion method can simply and efficiently culture primary rabbit BMECs with strong activity and high purity.

Sepsis is a common critical illness in clinical practice,characterized by rapid progression and high mortality.Its complex pathogenesis remains a major focus and challenge in the field of critical care medicine.Disseminated intravascular coagulation(DIC)is one of the most frequent and severe complications of sepsis,featuring systemic activation of the coagulation cascade and microthrombus formation,significantly increasing the mortality.Coagulation factorⅫ(FⅫ),a serine protease,is considered to have therapeutic potential for thrombosis without impairing normal hemostasis.Study reveal that neutrophil extracellular trap(NET),web-like DNA structures released through a unique process known as NETosis,provide negatively charged scaffolds that promote FⅫ binding and activation,thus triggering the intrinsic coagulation cascade and contributing to a hypercoagulable state.In recent years,increasing attention has been paid to the interaction between NET and FⅫ in sepsis complicated with DIC.These 2 factors play central roles in intravascular thrombus formation and coagulation activation.Beyond their antimicrobial function,NET can aggravate tissue injury and coagulation abnormalities by releasing proinflammatory mediators such as myeloperoxidase(MPO),neutrophil elastase(NE),and reactive oxygen species(ROS).FⅫ activation can further trigger the kallikrein-kinin system(KKS)and activate FⅪ,amplifying inflammation and thrombosis in a vicious cycle.Traditional Chinese medicine(TCM),as a key component of Chinese medical heritage,has demonstrated unique advantages in managing sepsis and its complications.Based on therapeutic principles such as"strengthening healthy qi and eliminating pathogenic factors"and"tonifying qi and activating blood circulation",TCM is believed to regulate immune function and correct coagulation disorders,thereby interfering with the hypercoagulable state mediated by NET and FⅫ,slowing the progression of DIC,and improving clinical outcomes.Several herbs,including Salvia miltiorrhiza,Astragalus membranaceus,Rheum officinale,Ligusticum chuanxiong,and Curcuma longa,have shown anticoagulant,antiplatelet,and anti-inflammatory properties.In addition,compound formulations such as Xuebijing injection and Qingwen Baidu decoction have demonstrated clinical efficacy in improving coagulation parameters,reducing D-dimer levels,and protecting organ function.Although current evidence on the effects of TCM on NET formation and FⅫactivation is still limited,its potential mechanisms and clinical value warrant further investigation.This review summarizes the critical roles and interplay of FⅫand NET in sepsis complicated with DIC and discusses the advances in TCM-based interventions,aiming to provide new perspectives for mechanism-oriented research and integrative therapeutic strategies.

Objectives:To establish an efficient in vitro culture method for spinal cord microvascular en-dothelial cells(SCMECs)of rabbits.Methods:15 Japanese large-ear white rabbits(2-week-old,either sex)were selected.Under sterile conditions,the spinal cord was obtained by incising the back skin,muscle layers and vertebral canal along its longitudinal axis.The spinal cord was minced,and myelin impurities were removed by density gradient centrifugation using bovine serum albumin.The spinal cord microvascular segments were obtained by filtration through a sieve.The enzymatic digestion method was used,with 0.1％type Ⅱ collage-nase digesting the microvascular segments for 20min,after which,the segments were seeded into culture flasks,and then placed in a CO2 incubator for static culture for 72h.The above procedures were performed with five rabbits at a time,repeated three times,and the target cells from the third culture were taken as the objects of observation and identification.The growth and morphological characteristics of the cells were ob-served under an inverted phase-contrast microscope.Immunocytochemical staining was used to detect the ex-pression of factor Ⅷ-related antigen to identify the cultured SCMECs.Results:Microscopically,the segments showed short-rod or branched shapes post-seeding.After 24h of culture,short spindle-shaped cells migrated out from around the microvascular segments and grew in a"budding"manner.After 48h,"island-like"cell colonies formed.After 72h,the cells covered the bottom of the dish,growing in a typical monolayer,"paving stone-like"mosaic pattern.Immunocytochemical staining showed that over 99％of the cells had brownish-red cytoplasm,with positive expression of factor Ⅷ-related antigen,hematoxylin stained the nuclei and was shown in blue.Conclusions:Enzymatic digestion can successfully isolate and culture SCMECs of Japanese large-ear white rabbits with good activity and high purity.