Effect of donepezil hydrochloride combined with memantine on cognitive function in patients with Alzheimer's disease / 中国基层医药

Objective:To analyze the effect of donepezil hydrochloride combined with memantine on cognitive function in patients with Alzheimer's disease (AD).Methods:A randomized controlled trial was conducted among 90 patients with AD who were treated at the Zaozhuang Mental Health Center from January 2021 to March 2023. The patients were divided into a donepezil group and a combination group using a random number table grouping method, with 45 patients in each group. The donepezil hydrochloride group received only oral administration of donepezil hydrochloride tablets, while the combination group received oral administration of both donepezil hydrochloride tablets and memantine tablets. The two groups were continuously treated for 12 weeks. Before and after treatment, the Activities of Daily Living scale (ADL) score, the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD) score, the Mini-Mental State Scale (MMSE) score, and biochemical indicators (homocysteine, neuron-specific enolase, and S100 β) were compared between the two groups. Adverse drug reactions were observed in each group.Results:After treatment, the ADL, BEHAVE-AD, and MMSE scores in the combination group were (78.9 ± 6.1) points, (5.2 ± 0.5) points, and (22.8 ± 2.2) points, respectively, and they were (65.2 ± 5.9) points, (9.6 ± 0.9) points, and (19.4 ± 2.4) points, respectively, in the donepezil hydrochloride group. The ADL and MMSE scores in the combination group were significantly higher than those in the donepezil hydrochloride group ( t = 10.83, 7.01, both P < 0.001). The BEHAVE-AD score in the combination group was significantly lower than that in the donepezil hydrochloride group ( t = -28.67, P < 0.001). After treatment, serum levels of homocysteine, neuron-specific enolase, and S100 β in the combination group were (17.8 ± 3.6) μmol/L, (16.8 ± 2.7) μg/L, and (17.4 ± 7.5) μg/L, respectively, which were significantly lower than (21.5 ± 3.3) μmol/L, (20.4 ± 3.7) μg/L, and (23.5 ± 5.1) μg/L in the donepezil hydrochloride group ( t = -5.08, -5.27, -4.51, all P < 0.001). The incidence of adverse drug reactions in the combination group was 13.3% (6/45), which was slightly, but not significantly, higher than 8.9% (4/45) in the donepezil group ( χ2 = 0.45, P = 0.502). Conclusion:The combination of donepezil hydrochloride and memantine can effectively improve the mental and behavioral symptoms and cognitive function of patients with AD, improve daily living ability, and do not increase adverse reactions. The combined therapy has high clinical application value.

Correlation between changes in homocysteine, uric acid, and lipid metabolism in patients with first-episode schizophrenia and their psychopathological status / 中国基层医药

Objective:To investigate the correlation between the changes in homocysteine, uric acid, and lipid metabolism and the psychopathological status of patients with first-episode schizophrenia.Methods:Between June 2021 and June 2023, a total of 82 patients with first-episode schizophrenia who received treatment at Wenzhou Seventh People's Hospital were included in the study group. Additionally, 82 individuals who underwent physical examination at the same hospital during the same period were included in the control group. The homocysteine levels were measured using the enzyme cycling method, while uric acid levels were determined via the uric acid enzyme method. Lipid metabolism levels were assessed using a fully automated biochemical analyzer. The positive and negative syndrome (PANSS) scale scores, which assess positive, negative, and general psychopathological status in patients with first-episode schizophrenia, were obtained. Serum homocysteine and uric acid levels, as well as lipid metabolism levels, were compared between the two groups. Pearson correlation analysis was used to assess the correlation between general psychopathological status scores and homocysteine, uric acid, and lipid metabolism levels. Multivariate logistic regression analysis was performed to investigate the factors (homocysteine, uric acid, or lipid metabolism levels) that affect first-episode schizophrenia.Results:The patients with first-episode schizophrenia exhibited positive symptom scores of (25.74 ± 5.63) points, negative symptom scores of (23.12 ± 6.76) points, general psychopathological status scores of (37.91 ± 5.87) points, and a total score of (86.77 ± 7.75) points. The serum homocysteine and uric acid levels in the study group were (23.12 ± 3.78) μmol/L and (345.12 ± 27.38) μmol/L, respectively. These values were significantly higher than those in the control group [(8.81 ± 2.19) μmol/L, (218.48 ± 23.51) μmol/L, t = 29.66, -31.78, both P < 0.05]. There were no significant differences in total cholesterol and triacylglycerol levels between the two groups (both P > 0.05). However, serum level of low density lipoprotein cholesterol in the study group was (3.26 ± 1.04) mmol/L, which was significantly higher than that in the control group [(2.34 ± 0.78) mmol/L, t = -6.41, P < 0.05]. Serum level of high density lipoprotein cholesterol in the study group was (1.02 ± 0.13) mmol/L, which was significantly lower than that in the control group [(1.17 ± 0.14) mmol/L, t = 7.11, P < 0.05). Pearson correlation analysis indicated a positive linear correlation between the general psychopathological status score and homocysteine, uric acid, and low density lipoprotein cholesterol levels. Conversely, it exhibited a negative linear correlation with high density lipoprotein cholesterol levels. Multivariate logistic regression analysis further showed that homocysteine, uric acid, and low density lipoprotein cholesterol levels were strongly associated with first-episode schizophrenia. Conclusion:In patients with first-episode schizophrenia, homocysteine, uric acid, and low density lipoprotein cholesterol levels are elevated, while high density lipoprotein cholesterol levels are reduced. There markers are closely related to patients' psychopathological status.

Effects of Zhuyu Pill （茱萸丸） and its Ingredient on Aortic Plaque and the NLRP3/Asc/Caspase-1 Signaling Pathway in Aortic Tissue of Mice in Atherosclerosis Model / 中医杂志

ObjectiveTo investigate the possible mechanism of action and combination of medicinals of Zhuyu Pill （茱萸丸） in the treatment of atherosclerosis. MethodsThirteen normal C57BL/6J mice were used as blank group， and 40 ApoE-/- mice of the same strain were randomly divided into model group， Huanglian （Coptis chinensis Franch.） group， Wuzhuyu （Tetradium ruticarpum） group， and Zhuyu Pill group， with 10 mice in each group. Except the blank group， the other groups were fed with high-fat diet for modeling， and the total modeling cycle was 12 weeks. After modelling， Huanglian group was given Huanglian solution 143.34 mg/（kg·d）， Wuzhuyu group with Wuzhuyu solution 301.78 mg/（kg·d）， Zhuyu Pill group with 261.07 mg/（kg·d） of Zhuyu Pill solution， blank group and model group were given 0.3ml of normal saline. Mice in all groups were gavaged for 12 weeks. The levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C） and fasting blood glucose were measured by biochemical method； fasting insulin in serum was detected by enzyme-linked immunosorbent assay （ELISA） and insulin resistance index was calculated； HE staining was used to observe the pathological and morphological changes of aortic tissue， liver and epididymal fat in mice， and oil red O staining was used to observe the lipid deposition of aortic tissue in mice， and the percentage of positive area of aortic HE staining， the percentage of positive area of aortic oil red O staining， the non-alcoholic fatty liver activity score （NAS） score， and the cross-sectional area of epididymal fat were calculated； serum levels of interleukin 1β （IL-1β） and interleukin 18 （IL-18） were detected by ELISA； protein expression of nucleotide-binding oligomerisation structural domain-like receptor family pyridoxal structural domain protein 3 （NLRP3） in aortic tissues was detected by Western blot； and immunofluorescence was used to detect the levels of apoptosis-associated granulocyte-like protein （ASC）， which contains the CARD structural domain （ASC） and cysteine-containing aspartic protease 1 （Caspase-1） protein expression； and real-time fluorescence PCR was used to detect NLRP3， ASC， Caspase-1， IL-1β， and IL-18 mRNA expression in mouse aortic tissues. ResultsCompared with blank group， serum TC， TG and LDL-C were increased， insulin was decreased， fasting blood glucose and insulin resistance index were increased in model group （P＜0.01）. Aortic plaque area increased， liver lipid deposition increased， epididymal fat cells volume increased， liver NAS score increased， and epididymal fat cross-sectional area increased. Serum IL-1β and IL-18 increased， and the expression levels of NLRP3， ASC， Caspase-1 protein and mRNA in aortic tissue increased （P＜0.01）. Compared with model group， serum TC， TG and LDL-C in Huanglian group， Wuzhuyu group and Zhuyu Pill group decreased， fasting insulin increased， fasting blood glucose and insulin resistance index decreased （P＜0.01）； aortic plaque area decreased significantly， liver lipid deposition decreased， liver NAS score decreased， epididymal fat cell volume decreased， epididymal fat cross section area decreased， serum IL-1β and IL-18 were decreased， the expression levels of NLRP3， ASC， Caspase-1 protein and mRNA in aortic tissue were decreased （P＜0.01）， and the improvement of all indexes in the Zhuyu Pill group was better than that in Huanglian and Wuzhuyu groups （P＜0.01）. ConclusionZhuyu Pill has a good therapeutic effect on atherosclerosis in mice， and the combination of Huanglian and Wuzhuyu has a synergistic effect， the mechanism of which may be related to regulation of the NLRP3/ASC/Caspase-1 aortic signaling pathway， and thus reducing the vascular inflammatory response.

Effect of Modified Shenhong Tongluo Prescription on Cell Apoptosis in Rats with Myocardial Ischemia-reperfusion Injury by Bcl-2/Bax/Caspase-3 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of modified Shenhong Tongluo prescription on cell apoptosis in rats with myocardial ischemia-reperfusion injury （MIRI）. MethodSixty Sprague-Dawley （SD） rats were randomly divided into a blank group， a model group， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription， and a simvastatin group. Except for the blank group， a rat model of MIRI was prepared by ligating the left anterior descending coronary artery. Starting from the first day after successful modeling， the blank group （1.0 mL·kg-1 physiological saline）， model group （1.0 mL·kg-1 physiological saline）， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription （1.031， 2.063， and 4.126 g·kg-1 Shenhong Tongluo prescriptiona standard concentrate）， and simvastatin group （0.71 mg·kg-1 simvastatin） were orally administered once daily for 2 weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of cardiomyocytes. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of serum creatine kinase isoenzyme （CK-MB）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α）. TdT-mediated dUTP nick-end labeling（TUNEL） staining was used to detect the apoptosis rate of rat cardiomyocytes. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and caspase-3. ResultCompared with the blank group， in the model group， HE staining showed disturbed arrangement of cardiomyocytes， incomplete fibers， focal necrosis of cardiomyocytes， and inflammatory cell infiltration； serum CK-MB， IL-6， and TNF-α levels were significantly increased （P<0.05）； apoptosis rate of cardiomyocytes was significantly increased （P<0.01）， with significantly increased expression levels of Bax and Caspase-3 proteins， and significantly decreased Bcl-2 expression （P<0.05）. Compared with the model group， the low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription significantly reduced CK-MB， IL-6， and TNF-α levels （P<0.05）， significantly downregulated cardiomyocyte apoptosis rate （P<0.05）， significantly decreased Bax and Caspase-3 proteins， and significantly increased Bcl-2 expression levels （P<0.01）. In the modified Shenhong Tongluo prescription groups， the expression levels of Bax and Caspase-3 proteins significantly decreased with increasing dosage， while the expression level of Bcl-2 significantly increased with increasing dosage of modified Shenhong Tongluo prescription （P<0.05）. ConclusionShenhong Tongluo prescription can alleviate myocardial tissue pathological damage and reduce myocardial cell apoptosis， possibly by inhibiting Caspase-3 and Bax expression and promoting Bcl-2 expression.

The expression of CRIP1 and STUB1 in cancer tissues of patients with hepatocellular carcinoma and their clinical prognostic significance / 国际检验医学杂志

Objective To investigate the expression of cysteine-rich intestinal protein 1(CRIP1),STIP1 ho-mology and U-box protein 1(STUB1)in cancer tissues of patients with hepatocellular carcinoma and their clinical prognostic significance.Methods From February 2018 to February 2020,112 patients with hepatocel-lular carcinoma were selected as the study objects.The expression of CRIP1 and STUB1 in cancer tissues and adjacent tissues of patients with hepatocellular carcinoma was detected by immunohistochemistry.To analyze the relationship between the expression of CRIP1 and STUB1 and their clinicopathological features in hepato-cellular carcinoma patients.Kaplan-Meier survival analysis of the effects of CRIP1 and STUB1 expression on the prognosis of patients with hepatocellular carcinoma.COX regression analysis was performed to analyze the prognostic factors of hepatocellular carcinoma.Results The positive rate of CRIP1 in cancer tissues of pa-tients with hepatocellular carcinoma was 62.50％(70/112),which was significantly higher than that in adja-cent tissues[7.14％(8/112)],the difference was statistically significant(x2=76.652,P＜0.05).The positive rate of STUB1 in cancer tissues of patients with hepatocellular carcinoma was 26.23％(32/112),significantly lower than that in adjacent tissues[82.14％(92/112)],and the difference was statistically significant(x2=73.284,P＜0.05).The expression of CRIP1 was negatively correlated with STUB1 in cancer tissues(r=-0.678,P＜0.001).There were significant differences in the positive rates of CRIP1 and STUB1 in hepato-cellular carcinoma patients with different TNM stages,histological grades and maximum tumor diameter(P＜0.05).The 3-year cumulative survival rate of CRIP1 positive group was significantly lower than that of CRIP1 negative group,with statistical significance(Log-rank x2=29.601,P＜0.001).The 3-year cumulative survival rate of STUB1 negative group was significantly lower than that of STUB1 positive group,with statistical sig-nificance(Log-rank x2=13.590,P＜0.001).TNM stage Ⅱ-Ⅲ,histological grade Ⅲ,maximum tumor diam-eter＞5 cm,CRIP1 positive and STUB1 negative were independent risk factors for prognosis of hepatocellular carcinoma patients.Conclusion CRIP1 expression is up-regulated and STUB1 expression is down-regulated in hepatocellular carcinoma tissues.The prognosis of patients with hepatocellular carcinoma can be evaluated clinically based on the expression of CRIP1 and STUB1 in hepatocellular carcinoma tissues.

Hei Xiaoyaosan Regulates Fas/FasL/Caspase-8/Caspase-3 Signaling Pathway to Inhibit Neuronal Apoptosis in AD Rats / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in regulating the tumor necrosis factor receptor superfamily member 6 （Fas）/Fas ligand （FasL）/cysteine protease-8 （Caspase-8）/cysteine protease-3 （Caspase-3） signaling pathway to intervene in neuronal apoptosis and prevent Alzheimer's disease （AD）. MethodNinety SPF-grade SD male rats of 4 months old were selected and randomly grouped as follows： 10 rats in the blank group， 10 rats in the sham group （bilateral hippocampus injected with 1 μL normal saline）， and 70 rats in the modeling group ［bilater hippocampus injected with 1 μL amyloid-beta protein 1-42 （Aβ1-42） solution for the modeling of AD］. Fifty successfully modeled rats were selected and randomly assigned into model， donepezil hydrochloride （0.45 mg·kg-1）， and high-， medium-， and low-dose （15.30， 7.65， 3.82 g·kg-1） Hei Xiaoyaosan groups. Rats were administrated with corresponding agents by gavage once a day for 42 days. Terminal-deoxynucleoitidyl transferase-mediated nick end labeling （TUNEL） was employed to observe the apoptosis of neurons in the cortex and hippocampus， and immunohistochemistry （IHC） was used to detect the expression of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to determine the mRNA levels of Fas， FasL， and Fas-associated protein with death domain （Fadd）. Western blot was used to determine the protein levels of Fas， FasL， Fadd， Caspase-3， cleved Caspase-3， Caspase-8， and cleved Caspase-8. ResultCompared with the blank group and sham group， the model group showed increased apoptosis rate in the cortex and hippocampus （P<0.01）， elevated Bax level （P<0.01）， lowered Bcl-2 level （P<0.01）， up-regulated mRNA levels of Fas， FasL， and Fadd in the hippocampus （P<0.01）， and up-regulated protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.01）. Compared with the model group， donepezil hydrochloride and Hei Xiaoyaosan at high and medium doses decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bax level （P<0.01）， elevated the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of Fas， FasL， and Fadd and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05， P<0.01） in the hippocampus. Low-dose Hei Xiaoyaosan decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of FasL and Fadd （P<0.05） and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05） in the hippocampus. ConclusionHei Xiaoyaosan can protect neurons in the cortex and hippocampus of AD rats by inhibiting the apoptosis mediated by the Fas/FasL/Caspase-8/Caspase-3 signaling pathway.

Broad-spectrum ginsentides are principal bioactives in unraveling the cure-all effects of ginseng

Stress and illness connection is complex and involves multiple physiological systems. Panax ginsengs, reputed for their broad-spectrum "cure-all" effect, are widely prescribed to treat stress and related illnesses. However, the identity of ginseng's "cure-all" medicinal compounds that relieve stress remains unresolved. Here, we identify ginsentides as the principal bioactives that coordinate multiple systems to restore homeostasis in response to stress. Ginsentides are disulfide-rich, cell-penetrating and proteolytic-stable microproteins. Using affinity-enrichment mass spectrometry target identification together with in vitro, ex vivo and in vivo validations, we show that highly purified or synthetic ginsentides promote vasorelaxation by producing nitric oxide through endothelial cells via intracellular PI3K/Akt signaling pathway, alleviate α1-adrenergic receptor overactivity by reversing phenylephrine-induced constriction of aorta, decrease monocyte adhesion to endothelial cells via CD166/ESAM/CD40 and inhibit P2Y12 receptors to reduce platelet aggregation. Orally administered ginsentides were effective in animal models to reduce ADP-induced platelet aggregation, to prevent collagen and adrenaline-induced pulmonary thrombosis as well as anti-stress behavior of tail suspension and forced swimming tests in mice. Together, these results strongly suggest that ginsentides are the principal panacea compounds of ginsengs because of their ability to target multiple extra- and intra-cellular proteins to reverse stress-induced damages.

Understanding the complexity of Tityus serrulatus venom: A focus on high molecular weight components

Tityus serrulatus scorpion is responsible for a significant number of envenomings in Brazil, ranging from mild to severe, and in some cases, leading to fatalities. While supportive care is the primary treatment modality, moderate and severe cases require antivenom administration despite potential limitations and adverse effects. The remarkable proliferation of T. serrulatus scorpions, attributed to their biology and asexual reproduction, contributes to a high incidence of envenomation. T. serrulatus scorpion venom predominantly consists of short proteins acting as neurotoxins (α and ß), that primarily target ion channels. Nevertheless, high molecular weight compounds, including metalloproteases, serine proteases, phospholipases, and hyaluronidases, are also present in the venom. These compounds play a crucial role in envenomation, influencing the severity of symptoms and the spread of venom. This review endeavors to comprehensively understand the T. serrulatus scorpion venom by elucidating the primary high molecular weight compounds and exploring their potential contributions to envenomation. Understanding these compounds' mechanisms of action can aid in developing more effective treatments and prevention strategies, ultimately mitigating the impact of scorpion envenomation on public health in Brazil.

Assessing proteases and enzymes of the trypanothione system in subpopulations of Leishmania (Viannia) braziliensis Thor strain during macrophage infection

BACKGROUND Leishmania (Viannia) braziliensis Thor strain exhibits a heterogeneous composition comprised of subpopulations with varying levels of infectivity. Clonal subpopulations were previously obtained from the strain Thor by sorting single-parasites and proceeding cultivation. The subpopulations used in this study are named Thor03, Thor 10 and Thor22. OBJECTIVES Phenotypic characteristics of the parasite, specially focusing on virulence factors and resistance to the antimicrobial mechanisms of macrophages, were investigate in these subpopulations. METHODS Cellular and molecular biology, as well as biochemistry approaches were applied to obtain the data analysed in this study. FINDINGS Relative quantification of gene expression was measured for calpain, cysteine protease B (CPB), and subtilisin proteases but no significant differences in these genes' expression among subpopulations was observed. However, subtilisin and CPB proteins were assessed as more abundant in Thor03 by fluorescence-labelled flow cytometry technique. Western Blotting assays, as semi-quantitative analysis in gel, showed higher concentrations of subtilisin (110 to 50 kDa) and CPB (40 to 18 kDa) in extract of intracellular amastigotes from subpopulations Thor03 and Thor10 and calpain (60 to 25 kDa) showed no significant differences among subpopulations. Complementary, higher trypanothione reductase activity was observed in Thor10 intracellular amastigotes and assays of susceptibility to hydrogen peroxide-inducing agents and nitric oxide donors conducted with promastigotes revealed greater resistance to in vitro oxidative stress induction for Thor10, followed by Thor03. MAIN CONCLUSIONS The data obtained for the virulence factors explored here suggest how multiple coexisting phenotypic-distinct subpopulations may contribute in adaptability of a single L. (V.) braziliensis strain during infection in the host cells.

Possible therapeutics for Pseudomyxoma peritonei: a rare, lethal, and the least investigated disease

Pseudomyxoma peritonei (PMP) refers to a growth disorder characterized by glycoprotein neoplasm in the peritoneum, where mucin oversecretion occurs. The tumors of the appendix region are well associated with PMP; however, ovarian, colon, stomach, pancreas, and urachus tumors have also been linked to PMP. Other mucinous tumors in the pelvis, paracolic gutters, greater omentum, retrohepatic space, and Treitz ligament can be the reason for PMP. Despite being rare and having a slow growth rate, PMP can be lethal without treatment. It is treated with neoadjuvant chemotherapy with the option of cytoreductive surgery and intraperitoneal chemotherapy. In the current study, we hypothesize that there may be novel gentle ways to inhibit or eliminate the mucin. Dr. David Morris has used mucolytics - such as bromelain and N-acetyl cysteine to solubilize mucin. In the present review, we aimed to study the regulation of mucin expression by promoter methylation, and drugs that can inhibit mucin, such as boldine, amiloride, naltrexone, dexamethasone, and retinoid acid receptors antagonist. This review also explored some possible pathways, such as inhibition of Na + , Ca2+ channels and induction of DNA methyltransferase along with inhibition of ten-eleven translocation enzymes, which can be good targets to control mucin. Mucins are strong adhesive molecules that play great roles in clinging to cells or cell to cell. Besides, they have been greatly involved in metastasis and also act as disease markers for cancers. Diagnostic markers may have exclusive roles in disease initiation and progression. Therefore, the present review explores various drugs to control and target mucin in various diseases, specifically cancers. (AU)

Babaodan Alleviates APAP-induced Acute Liver Injury in Mice by Inhibiting NLRP3/Caspase-1 Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect of Babaodan （BBD） on the NOD-like receptor pyrin domain containing 3/cysteine aspartate-specific protease-3 （NLRP3/Caspase-1） pathway proteins in mice with acetaminophen （APAP）-induced acute liver injury. MethodC57BL/6 mice were randomly grouped， and BBD （75， 150， 300 mg·kg-1， ig） was administered twice a day for three days. After 2 hours of the last administration， the mice were treated with APAP （400 mg·kg-1， ip）， and the eyeballs were removed to collect blood after 14 hours. Then they were sacrificed by cervical dislocation for sample collection. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of liver tissue cells， and biochemical methods were used to detect the activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， superoxide dismutase （SOD）， malondialdehyde （MDA） and myeloperoxidase （MPO） in serum of mice in each group. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6， and Western blot was performed to determine the protein expression of cyclooxygenase-2 （COX-2）， inducible nitric oxide synthase （iNOS）， NLRP3， Caspase-1 and IL-18 in the liver of mice. ResultCompared with the conditions in normal group， the hepatic lobule structure of mice in the model group was partially destroyed， and the hepatic sinusoids were dilated. And the expression levels of ALT and AST in serum， the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the mRNA levels of IL-1β， IL-6 and TNF-α were increased （P<0.05， P<0.01）. Compared with the model group， the administration groups had improvement in liver cell rupture and hepatic sinusoidal compression， and a dose-dependent decrease in the levels of ALT and AST in serum as well as the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the the mRNA levels of IL-1β， IL-6 and TNF-α in liver tissue （P<0.05， P<0.01）. ConclusionBBD can reduce APAP-induced acute liver injury in mice. The mechanism may be related to anti-oxidative stress， inhibition of NLRP3/Caspase-1 pathway， and decreased expression levels of IL-1β， IL-18， TNF-α and IL-6.

Study on the role and mechanism of osteogenesis induced by advanced platelet-rich fibrin and β-tricalcium phosphate complex / 口腔疾病防治

Objective @#To investigate the role and mechanism of bone formation caused by the ratio of advanced platelet-rich fibrin (A-PRF) and β-tricalcium phosphate (β-TCP) in rabbit femur defect model, which provides a new idea for clinical treatment of bone defect.@*Methods @#Twenty-four New Zealand white rabbits were divided into model group, 1∶1 complex group (A-PRF∶β-TCP=1∶1), 2∶1 complex group (A-PRF∶β- TCP=2∶1) and 4∶1 complex group (A-PRF∶β- TCP=4∶1）, with 6 rabbits in each group. Femoral defect models were constructed in each group. In the composite group, the bone defect was filled with composite material, while in the model group, no material was filled. After 8 weeks, the animals were euthanized and specimens were collected. Bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.SP) and trabecular number (Tb.N) in femoral defect tissue were measured by micro-CT and photographed. Hematoxylin - eosin staining was used to detect the pathological changes of new bone tissue. The morphological changes of the new bone tissue were observed by scanning electron microscopy. Determination of phospho-mitogen activated protein kinase p38 (p-p38MAPK), CCAAT/enhancer binding protein homologous protein (CHOP) and phospho-cysteine aspartic protease-3 (p-Caspase3) in newborn femur by ELISA. The mRNA expressions of osteoprotegerin (OPG), bone morphogenetic protein-2 (BMP-2), receptor activator of nuclear factor kappa-B ligand (RANKL) and p38MAPK were detected by real-time quantitative PCR. The expression of OPG, BMP-2, RANKL, p-p38MAPK and p-Caspase3 protein in the new bone tissue was observed by immunohistochemistry. @*Results @#In the model group, bone formation in the femoral defect area was slow and osteogenic quality was poor. Compared with the model group, the bone formation and neocapillaries of femoral defect area in the complex group was good, BMD, BV.TV, Tb.Th, Tb.N were increased, and Tb.Sp were decreased, the expressions of p-p38MAPK, CHOP and p-Caspase3 were decreased, and the mRNA and protein expressions of OPG and BMP-2 were increased. The mRNA expression of RANKL and p38MAPK was decreased. Apoptosis in new bone tissue of each group showed the lowest apoptosis rate in samples of the 2∶1 complex group (P<0.05); A-PRF: β-TCP=2∶1 ratio has the best osteogenic effect. @*Conclusion@#The complex composed of A-PRF and β-TCP can promote the expression of OPG, inhibit the expression of RANKL and phosphorylation of p38MAPK, reduce the apoptosis of new bone tissue cells, and promote osteogenic differentiation.

Relationship between preoperative serum cystatin C concentration and postoperative delirium / 中华麻醉学杂志

Objective:To evaluate the relationship between preoperative serum cystatin C (Cys C) concentration and postoperative delirium (POD) in the patients.Methods:Three hundred and ninety patients, aged >50 yr, of American Society of Anesthesiologists Physical Status classification Ⅰor Ⅱ, scheduled for elective knee and hip replacement under combined spinal-epidural anesthesia, with Mini-Mental State Examination scores >23 at 1 day before operation, were included in the study. Peripheral blood samples were collected before operation, and the serum Cys C concentration was measured by the latex-enhanced immunoturbidimetric assay. The cerebrospinal fluid (CSF) 2 ml was collected after successful spinal-epidural puncture for determination of amyloid-β 42 (Aβ42), total tau (T-tau) and phosphorylated tau (P-tau) concentrations by enzyme-linked immunosorbent assay. The patients were divided into POD group and non-POD group according to whether POD occurred. The logistic regression analysis was used to identify the risk and protective factors for POD, and the mediating effect of CSF biomarkers was analyzed. The receiver operating characteristic curve was drawn to evaluate the accuracy of serum Cys C concentration and combination of serum Cys C conceatration and CSF biomarker concentration in predicting POD.Results:Three hundred and twenty-seven patients were finally enrolled, and the incidence of POD was 13.5%. The results of logistic regression showed that increased serum Cys C concentration and increased concentrations of P-tau and T-tau in CSF were risk factors for POD, while increased concentration of Aβ42 and increased Aβ42/P-tau ratio and Aβ42/T-tau ratio in CSF were protective factors for POD ( P<0.05) after adjusting for multiple confounding variables such as age, sex, years of education, Mini-Mental State Examination score, smoking history, drinking history, hypertension and diabetes history. The mediation analysis showed that the relationship between serum Cys C concentration and POD was mediated by T-tau concentration in CSF (11.1%) and by Aβ42/T-tau ratio in CSF (18.0%). The area under the receiver operating characteristic curve of serum Cys C and CSF biomarker concentrations in predicting POD was 0.807 ( P<0.001). Conclusions:Increase in preoperative serum Cys C concentration is a risk factor for POD. T-tau concentration and Aβ42/T-tau ratio in CSF serve as the key mediators in the relationship between preoperative serum Cys C concentration and POD.

Relationship between serum CASP2 expression and NBNA score and prognosis in children with hypoxic-ischemic encephalopathy / 国际检验医学杂志

Objective To study the correlation between serum cysteine aspartate proteinase 2(CASP2)level and neonatal behavioral neurological assessment(NBNA)score in children with hypoxic-ischemic en-cephalopathy(HIE),and the value of CASP2 in evaluating prognosis.Methods A total of 114 neonates with HIE in this hospital from January 2018 to April 2022 were selected as HIE group.According to the severity of the disease,the neonates were divided into mild group(40 cases),moderate group(52 cases)and severe group(22 cases).According to the prognosis after 6 months of treatment,114 neonates with HIE were divided into poor prognosis group(26 cases)and good prognosis group(88 cases).At the same time,106 healthy neonates born in the hospital were selected as the control group.The NBNA score was recorded,and the serum CASP2 level was detected by enzyme-linked immunosorbent assay.Pearson correlation was used to analyze the corre-lation between serum CASP2 level and NBNA score in HIE children.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of NBNA score and serum CASP2 level for the prognosis of ne-onates with HIE.Results The level of serum CASP2 in HIE group was higher than that in control group,and the NBNA score was lower than that in control group(P<0.05).The level of serum CASP2 in the severe group was higher than that of the moderate group and mild group,and the NBNA score was lower than that of moderate group and mild group(P<0.05).The level of serum CASP2 in the moderate group was higher than that of the mild group,and the NBNA score was lower than that of the mild group(P<0.05).The NBNA score was negatively correlated with the serum level of CASP2 in the children with HIE(r=-0.608,P<0.05).The poor prognosis group had a significantly higher serum level of CASP2 and a significantly lower NBNA score than the good prognosis group(P<0.05).The serum level of CASP2 and NBNA score separate and joint area under the curve of the poor children with HIE prognosis were 0.875(95%CI 0.797-0.952),0.748(95%CI 0.639-0.857),0.927(95%CI 0.873-0.981).Conclusion The level of serum CASP2 increases with the severity of HIE,and is closely related to the NBNA score,which has an important prognostic value.

Mechanism of action and clinical significance of CST1 in the progression of gastric cancer / 国际肿瘤学杂志

Cysteine protease inhibitor 1 (CST1) is a member of type 2 Cystatins superfamily, plays a key role in targeted regulation. CST1 is highly expressed in gastric cancer, promotes tumor cell migration and invasion by activating the epithelial-mesenchymal transformation pathway and Wnt pathway, and regulates tumor growth and progression in combination with the corresponding target genes of homeobox C10 and glutathione peroxidase 4. A deeper understanding of the role and function of CST1 in gastric cancer will help to further develop potential therapeutic targets and diagnostic and prognostic markers for gastric cancer.

Effect and mechanism of hyperthermia on cell apoptosis of laryngeal squamous cell carcinoma patients / 中华放射肿瘤学杂志

Objective:To evaluate the effect of hyperthermia on the apoptosis and the expression levels of cysteine-containing aspartate-specific protease-3 (Caspase-3) and phosphorylated protein kinase B (p-AKT) in laryngeal squamous cell carcinoma cells.Methods:A prospective study was conducted on 30 patients with laryngeal squamous cell carcinoma who were treated at the First Affiliated Hospital of Zhengzhou University from October, 2021 to October, 2022. Three times of hyperthermia were performed with a time interval of 24 h. The tumor tissue samples were collected from 30 patients before and after hyperthermia and divided into before hyperthermia group (group A ) and after hyperthermia group (group B). Self-control study mode was adopted for comrparative analysis. The cell apoptosis was detected by TUNEL assay. The expression levels of Caspase-3 and p-AKT in the tissues were detected by immunohistochemistry. Positive cell ratio and immunohistochemistry (IHC) score were recorded. Comparison between two groups was performed by paired t-test. The correlation between the degree of apoptosis and the changes of Caspase-3 and p-AKT molecules was assessed by Pearson correlation analysis. Results:No evident adverse reactions were observed in 30 patients after hyperthermia. The apoptosis index of laryngeal squamous cell carcinoma cells in group A was 2.37%±1.33%, and 4.27%±3.93% in group B ( P=0.006). In group A, the ratio of Caspase-3 positive cells in tumor tissues was 62.31%±19.49% and 80.79%±17.15% in group B ( P=0.001). The ratio of p-AKT positive cells in group A was 31.26%±19.30%, and 26.26%±15.86% in group B ( P=0.023). There was a positive correlation between the degree of apoptosis and the changes of Caspase-3 molecule ( r=0.544, P=0.002), but a negative correlation was noted between the degree of apoptosis and the changes of p-AKT molecule ( r=-0.434, P=0.017). Conclusion:Hyperthermia can promote the apoptosis of tumor cells in laryngeal squamous cell carcinoma, which may be related to Caspase-3 dependent apoptosis, and the inhibition of AKT phosphorylation is also involved in this process.

Jiedu Huoxue Prescription Regulates NF-κB/NLRP3/Caspase-1 to Promote Vascular Reendothelialization in Rat Model of Injured Thoracic Aorta / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Jiedu Huoxue prescription in promoting the reendothelialization of injured vessels by regulating the nuclear factor （NF）-κB/NOD-like receptor protein 3 （NLRP3）/cysteine-aspartic acid protease （Caspase）-1-mediated pyroptosis. MethodA rat model of injured thoracic aorta was established by balloon injury， and 36 rats were assigned into shame surgery， model， low-， medium-， and high-dose Jiedu Huoxue prescription， and atorvastatin calcium tablet groups. The injured aortic segment was collected 28 days after surgery. Hematoxylin-eosin （HE） staining and Evans blue staining were conducted to reveal the changes of vascular structural morphology and the reendothelialization of blood vessels， respectively. The enzyme-linked immunosorbent assay （ELISA） was employed to determine the levels of tumor necrosis factor-α （TNF-α）， intercellular adhesion molecule-1 （ICAM-1）， interleukin （IL）-1β， and nitric oxide （NO） in the serum. Western blotting was employed to determine the expression of endothelial nitric oxide synthase （eNOS）， NF-κB p65， phospho-NF-κB p65 （p-NF-κB p65）， NLRP3， and Caspase-1 in the vascular tissue. ResultThe model group showed thickened endovascular membrane， proliferation and disarrangement of smooth muscle cells of the artery wall， obvious inflammatory cell infiltration， and narrowed luminal area. Jiedu Huoxue prescription and atorvastatin calcium tablets mitigated the pathological changes of the thoracic aorta in different degrees. After balloon injury， the endothelial coverage rate of the model group decreased significantly， while Jiedu Huoxue prescription and atorvastatin calcium tablets increased the reendothelialization rate （P<0.05）. Compared with the shame surgery group， the model group showed elevated levels of TNF-α， ICAM-1， and IL-1β （P<0.01） and lowered NO level （P<0.01） in the serum. In addition， the model group presented down-regulated protein level of eNOS （P<0.01） and up-regulated phosphorylation of pyroptosis-associated proteins NLPR3， Caspase-1， and NF-κB p65 in the vascular tissue （P<0.05， P<0.01）. Compared with the model group， Jiedu Huoxue prescription and atorvastatin calcium tablets lowered TNF-α， ICAM-1， and IL-1β levels （P<0.05， P<0.01） and elevated the NO level in the serum （P<0.05， P<0.01）. Moreover， the drugs up-regulated the expression of eNOS （P<0.01） and down-regulated the expression of NLRP3， Caspase-1， and NF-κB p65 （P<0.05， P<0.01） in the vascular tissue. ConclusionJiedu Huoxue prescription can promote the reendothelialization and inhibit the intimal hyperplasia of vessels after balloon injury by regulating the NF-κB/NLRP3/Caspase-1 pathway to inhibit pyroptosis and reduce endothelial inflammatory injury.

Mechanism of miR⁃148b⁃3p regulating proliferation of keloid derived fibroblasts / 安徽医科大学学报

Objective @#To analyze the effect of miR⁃148b⁃3p on the proliferation of keloid derived fibroblasts. @*Methods @#The expression levels of miR⁃148b⁃3p and SPARC in human keloid derived fibroblasts (HKF) and normal human fibroblasts (NFS) were analyzed by real time PCR. The expression level of SPARC protein was detected by Western blot. The effects of miR⁃148b⁃3p and SPARC on HKF proliferation were analyzed by CCK⁃8 method the luciferase reporter plasmid was constructed. The targeted binding site of miR⁃148b⁃3p and the target gene was analyzed by luciferase reporter gene method.@*Results @#miR⁃148b⁃3p was low expressed in HKF and SPARC was high expressed in HKF. Transfection of miR⁃148b⁃3p in HKF cells could down regulate the expression of SPARC and inhibit cell proliferation. Online analysis software predicted that miR⁃148b⁃3p could target the 3 ′⁃ UTR binding SPARC ; The results of dual luciferase reporter gene further confirmed that miR⁃148b⁃3p could target the 3 ′⁃ UTR of SPARC. Transfection of SPARC eukaryotic expression plasmid into HKF transfected with miR⁃148b⁃3p could counteract the effect of miR⁃148b⁃3p and restore cell proliferation. @*Conclusion @# miR⁃148b⁃3p can inhibit the proliferation of HKF by targeting the 3 ′⁃ UTR of SPARC and inhibiting its expression.

Recent advance in mechanism of Hevin/SPARC in central nervous system synaptogenesis/reorganization / 中华神经医学杂志

Hevin is one of the extracellular matrix proteins secreted by astrocytes. Under physiological conditions, Hevin plays an important role in synaptogenesis in the central nervous system (CNS); secreted protein, acidic and rich in cysteine (SPARC) is its homologue and antagonizes the synaptogenic effects of Hevin. In pathological conditions, the expressions of Hevin and SPARC are altered, suggesting their possible roles at synaptic reorganization in various disease process, such as brain injury, Alzheimer's disease, epilepsy and brain tumors; however, the specific mechanism is not totally understood yet. So this paper reviews the mechanism of Hevin/SPARC in CNS synaptogenesis, reorganization and diseases to provide ideas for further research.

Effects of ICMT gene silencing on the invasion and migration of human salivary adenoid cystic carcinoma cells in vitro / 口腔疾病防治

Objective @#To investigate the effect of isoprene cysteine carboxymethyltransferase (ICMT) gene on the migration and invasion of salivary adenoid cystic cancer cells (SACC) and the related mechanism, to provide experimental evidence for molecular targeted therapy of SACC.@*Methods@# Adenoid cystic cancer cells SACC-LM and SACC-83 were cultured in vitro, and siRNA was transfected into human SACC-LM and SACC-83 cells (experimental group) by transient transfection of a liposome vector. A blank control group and negative control group were set up respectively (transfected NC-siRNA). qRT-PCR was peformed to measure the mRNA expression of ICMT and RhoA in each group after transfection and to determine the silencing efficiency. The expression of ICMT, membrane RhoA, total RhoA, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and Rho associated with coiled helical binding protein kinase 1 (ROCK1) in each group was detected by Western blot. The proliferation abilityies of SACC cells was detected by CCK-8 assay. The migration and invasion ability of SACC cells were detected by comparing the relative healing area of cell scratch assay and the number of Transwell assay cells. @*Results@#After transfection of ICMT-siRNA into SACC-LM and SACC-83 cells, the expression of ICMT gene and protein in the experimental group was significantly decreased compared with the negative control group and blank control group (P<0.05), but there were no significant differences in the expression of RhoA gene and total protein among all groups (P>0.05). The expression of RhoA membrane proteins, ROCK1, MMP-2, MMP-9 in the experimental group was significantly decreased compared with that in the negative control group and blank control group (P<0.05). Cell proliferation ability was significantly decreased (P<0.05). The migration and invasion abilities were significantly decreased (P<0.05). @*Conclusion @#In vitro silencing of ICMT gene can effectively inhibit the migration and invasion of human SACC-LM and SACC-83 cells, and the mechanism may be related to RhoA-ROCK signaling pathway.