Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method to simultaneously determine 10 main components, including berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in Compound Dihuang oral solution. Methods An UPLC-MS/MS method was established with an ACQUITY UPLC BEH-C18 （2.1 mm×100 mm, 1.7 μm）column and mobile phase of 0.1% formic water（A）-methanol solution（B） in a gradient elution manner. The flow rate of mobile phase was 0.2 ml/min.The temperature of column was 30℃. The injection volume was 2 μl. The MS detection was in MRM mode. Results 10 components in Compound Dihuang oral solution had a good linear relationship within their concentration range,and the precision, repeatability, stability and recovery met the requirements. The contents of berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in 7 batches of samples were （89.7-95.6） μg/ml, （164.0-177.7） μg/ml, （540.0-610.0） μg/ml, （408.7-429.0） μg/ml, （726.0-825.0） μg/ml, （503.7-572.0） μg/ml, （1380.0-1540.0） μg/ml, （2596.7-2896.7) μg/ml, （4866.7-5520.0) μg/ml, （61.8-67.2) μg/ml, respectively. Conclusion The established method was easy to be repeated and operated. The contents of 10 components in Compound Dihuang oral solution could be determined quickly and accurately, which provided a reference for the quality control of hospital preparation.

[摘 要] 目的：探讨RNA结合蛋白15（RBM15）通过三磷酸腺苷酶家族蛋白3A（ATAD3A）调控Wnt/β-catenin通路对宫颈癌细胞恶性生物学行为的影响。方法：利用TCGA数据库分析宫颈癌组织中RBM15 mRNA表达水平及其与患者预后的关系。收集2024年1月至10月期间在赣州市人民医院手术切除的32例宫颈癌组织及癌旁组织标本，以及宫颈癌细胞HeLa、MS-751、C-33A和SiHa，通过免疫组化、WB法检测宫颈癌组织和细胞中RBM15的表达水平。利用SRAMP在线数据库筛选ATAD3A mRNA的m6A修饰位点。通过RNA免疫沉淀实验、RNA衰变实验及挽救实验鉴定RBM15与ATAD3A mRNA的相互作用。采用RNA干扰技术和病毒感染技术，在宫颈癌HeLa和SiHa细胞敲低或过表达RBM15和ATAD3A，qPCR和WB法检测mRNA和蛋白表达，CCK-8法、划痕实验和Transwell实验检测各组细胞的增殖、迁移和侵袭能力。结果：宫颈癌组织中RBM15 mRNA和蛋白阳性率均显著高于癌旁组织（均P < 0.001）。宫颈癌HeLa、MS-751、C-33A和SiHa细胞RBM15蛋白表达水平显著高于正常宫颈细胞Ect1/E6E7（均P < 0.01）。RBM15 mRNA高表达组患者5年无进展生存率低于低表达组患者（P < 0.01）。宫颈癌组织中ATAD3A的表达水平显著高于癌旁组织（P < 0.001），RBM15 mRNA与ATAD3A mRNA呈正相关（r = 0.601，P < 0.05）。ATAD3A mRNA的501、5 312、12 137位点存在高可信度的m6A修饰位点。HeLa、SiHa细胞中过表达RBM15后，ATAD3A mRNA和蛋白表达升高，而敲低RBM15后，ATAD3A mRNA和蛋白表达降低（均P < 0.001）。RNA免疫沉淀实验显示，与IgG组相比，RBM15抗体的免疫沉淀中ATAD3A mRNA显著富集（均P < 0.001）。MeRIP-qPCR实验显示，ATAD3A mRNA 501、5 312、12 137位点均存在明显的m6A甲基化富集（均P < 0.001）。RNA衰变实验显示，敲低RBM15能降低HeLa、SiHa细胞ATAD3A mRNA的半衰期和稳定性（均P < 0.001）。敲低HeLa、SiHa细胞RBM15的表达，显著抑制癌细胞的增殖、迁移及侵袭，Wnt/β-catenin通路相关蛋白Wnt3、β-catenin、vimentin表达均显著降低（均P < 0.001）；而过表达ATAD3A可完全逆转上述抑制作用（均P < 0.001）。结论：RBM15通过m6A修饰ATAD3A mRNA调控Wnt/β-catenin信号通路，从而促进宫颈癌细胞增殖、迁移及侵袭。

Vascular damage plays a significant role in the onset and progression of Alzheimer's disease (AD). However, the precise molecular mechanisms underlying the induction of neuronal injury by vascular damage remain unclear. The present study aimed to examine the impact of fibrinogen (Fg) on tau pathology. The results showed that Fg deposits in the brains of tau P301S transgenic mice interact with tau, enhancing the cytotoxicity of pathological tau aggregates and promoting tau phosphorylation and aggregation. Notably, Fg-modified tau fibrils caused enhanced neuronal apoptosis and synaptic damage compared to unmodified fibrils. Furthermore, intrahippocampal injection of Fg-modified tau fibrils worsened the tau pathology, neuroinflammation, synaptic damage, neuronal apoptosis, and cognitive dysfunction in tau P301S mice compared to controls. The present study provides compelling evidence linking Fg and tau, thereby connecting cerebrovascular damage to tau pathology in AD. Consequently, inhibiting Fg-mediated tau pathology could potentially impede the progression of AD.
Animals ; tau Proteins/metabolism* ; Alzheimer Disease/metabolism* ; Fibrinogen/metabolism* ; Mice, Transgenic ; Mice ; Disease Models, Animal ; Memory Disorders/metabolism* ; Male ; Mice, Inbred C57BL ; Brain/metabolism* ; Hippocampus/metabolism* ; Protein Aggregation, Pathological/metabolism* ; Apoptosis ; Phosphorylation

Objective:To explore the association between cardiovascular health behaviors and hyperuricemia (HUA) among community residents of different age groups.Methods:It was a cross-sectional study. A total of 2 138 community residents aged (47.1±11.6) years with 1 012 males (47.3%) were selected by cluster sampling method as study subjects in Taiyuan Nanzhai Community from March to November 2020. There were 104 cases aged 8-18 years (underage group), 868 cases aged 19-44 years (youth group), 625 cases aged 45-59 years (middle-aged group), 375 cases aged 60-74 years (young elderly group), and 166 cases aged≥75 years (elderly group). Blood uric acid was measured and>420 μmol/L was defined as HUA, there were 385 cases with high uric acid level (HUA group) and 1 753 cases with normal uric acid levels (control group). The general information was collected by questionnaires, and general ergonomic indicators were measured on-site by medical personnel. The cardiovascular health behaviors included smoking, exercise, diet, and sleep in study subjects were documented. Multivariate logistic regression model was used to analyze the related factors of HUA.Results:Among 2 138 participants, 1 161 (54.3%) had never smoked (up to standard), and the order of proportion of non-smokers from high to low was underage group, elderly group, young elderly group, young group and middle-aged group ( P<0.001); 486(22.7%) people met the dietary standards, and the order of proportion of meeting the dietary standards from high to low was underage group, elderly group, young elderly group, middle-aged group, and young group ( P<0.001); 554(25.9%) people achieved physical fitness standards, and the order of the proportion of achieving physical fitness standards from high to low was young elderly group, middle-aged group, elderly group, underage group, and young group ( P<0.001); 783 (36.6%) people met the sleep standards, and the order of proportion meeting the sleep standards from high to low was underage group, youth group, middle-aged group, young elderly group, and elderly group ( P<0.001). Among the participants, only 39 (1.8%) met the standards for all 4 types of behaviors; 485 (22.7%), 1 229 (57.5%), and 424 (19.8%) had low, medium, and high total scores for ideal cardiovascular health behaviors, respectively. The total score of ideal cardiovascular health behaviors ranked from high to low was elderly group, young elderly group, underage group, middle-aged group, and young group ( P<0.001). The multivariate logistic regression analysis showed that body mass index (BMI) ( OR=1.125, 95% CI: 1.086-1.165) was positively correlated with HUA, while female ( OR=0.241, 95% CI: 0.182-0.320), total score of cardiovascular health behaviors (compared to lower level, intermediate level: OR=0.186, 95% CI: 0.127-0.273, high level: OR=0.038, 95% CI: 0.020-0.072), and number of achieved healthy behavior items ( OR=0.757, 95% CI: 0.621-0.922) were negatively correlated with HUA. Conclusions:The underage group has the highest number of people who meet the smoking, dietary, and sleep standards, the young and elderly group has the highest number of people who meet the exercise standards, and the elderly group has the highest total score for ideal cardiovascular health behaviors. There is a positive correlation between BMI and HUA, while there is a negative correlation of HUA with female, the total score and number of achieved items of cardiovascular health behaviors.

Objective To analyze the epidemiological characteristics of an outbreak of carbapenem resistant Klebsiella pneumoniae(CRKP)hospital infection in the neonatal intensive care unit(NICU),and to explore possible causes of the outbreak.Methods Case analysis,epidemiological studies and environmental detection methods were used to investigate the detection of CRKP in the NICU of a hospital from March 11,2022 to May 3,2022,and the causes and corresponding prevention and control measures were analyzed.Results This CRKP outbreak in the NICU occurred in two phases.The first phase was from March 11,2022,to March 30,2022,during which 3 people contracted CRKP hospital infections.The second phase was from April 13,2022,to May 3,2022,during which 4 people contracted CRKP hospital infections.The source of the hospital infection outbreak was patient No.1.All 13 CRKP cases showed sensitivity only to amikacin and tigecycline,with intermediate sensitivity to levofloxacin,and high resistance to other antibiotics.All 13 CRKP cases carried resistant genes for metalloenzymes.After the outbreak,the ward environment was sampled three times,with 35 samples taken in total,and 14 of them tested positive for CRKP.After strict environmental cleaning,disinfection,and multiple control measures,no new hospital infection cases similar to this outbreak were detected after May 13,2022.Conclusion The outbreak of NICU hospital infections may be due to the contamination of the ward environment,medical equipment,and inadequate hand hygiene of medical staff,leading to its spread.Therefore,strict cleaning and disinfection of the environmental surfaces,strengthening hand hygiene of medical staff,and implementing multiple measures can effectively control hospital infections of CRKP.

Abdomen/diagnostic imaging* ; Artificial Intelligence ; Body Composition ; Magnetic Resonance Imaging ; Practice Guidelines as Topic

Objective:To determine mutations in the PTPN11 gene in a family with LEOPARD syndrome.Methods:Clinical evaluation was carried out in a large pedigree with confirmed LEOPARD syndrome diagnosed in Hwa Mei Hospital, University of Chinese Academy of Sciences. Peripheral blood samples were obtained from 4 patients and 2 unaffected healthy members in the family, as well as 100 unrelated healthy controls. DNA was extracted from the blood samples, and PCR was performed to amplify all exons of the PTPN11 genes, followed by Sanger sequencing.Results:There were 14 members in 3 generations of the family, 6 of whom were affected (3 males and 3 females) , demonstrating an autosomal dominant inheritance pattern. Skin lesions were mainly distributed on the face, trunk and limbs, accompanied by special facial features and cardiovascular system abnormalities. A missense mutation c.1632G>T (p.R558L) in the PTPN11 gene was identified in the 4 patients, which resulted in the substitution of arginine by leucine at amino acid position 558. This mutation had not yet been reported previously. No mutation was detected in the PTPN11 gene in the 2 unaffected family members or 100 healthy controls.Conclusion:The missense mutation c.1632G>T in exon 13 of the PTPN11 gene may be the molecular basis for LEOPARD syndrome in this family.

Objective To investigate the correlations of P2Y12 gene polymorphisms with clopidogrel resistance and long-term outcome in patients with acute ischemic stroke. Methods From June 2015 to June 2017, consecutive patients with acute ischemic stroke admitted to the Department of Neurology, Nanjing Jiangbei People's Hospital were enrolled. Thromboelastography was used to measure platelet inhibition rate and assess clopidogrel resistance. Polymerase chain reaction was used to assay C34T and G52T polymorphisms of P2Y12 gene. The patients were followed up at 12 months after discharge. The primary outcome was combined outcome of stroke recurrence, myocardial infarction, and death due to cardiocerebrovascular events. Results A total of 214 patients were enrolled, 51 (23.8%) had clopidogrel re-sistance and 29 (13.4%) had major outcome events. One hundred twenty-eight (59.8%) patients were C34T CC genotype and 86 (40.2%) were CT+TT genotype. The proportion of clopidogrel resistance in patients with CT+TT genotype was significantly higher than that with CC genotype ( 76.5% vs.28.8%;χ2=25.672, P=0.001). There were 131 patients (61.2%) with G52T GG genotype and 83 (38.8%) with GT+TT genotype. There was no significant difference in the proportion of clopidogrel resistance between the GT+TT genotype and the GG genotype (43.1% vs.37.4%; χ2=0.534, P=0.465). Multiple logistic regression analysis indicated that age (odds ratio [OR] 1.064, 95%confidence interval [CI] 1.009-1.115;P=0.021), diabetes ( OR 3.773, 95%CI 1.672-8.475; P=0.004), and C34T CT+TT genotype ( OR 9.087, 95%CI 4.416-22.665; P=0.002) were the independent risk factors fot clopidogrel resistance. Cox proportional hazards model analysis showed that age (Hazard ratio [HR] 1.058, 95%CI 1.001-1.121; P=0.049), hypertension ( HR 3.105, 95%CI 1.149-9.523; P=0.028), homocysteine ( HR 1.101, 95%CI 1.020-1.190; P=0.014), and C34T CT+TT genotype ( HR 2.588, 95%CI 1.121-5.967; P=0.026) were independent risk factors for the composite outcome. Conclusion C34T polymorphism of P2Y12 gene in patients with acute ischemic stroke may be a risk factor for clopidogrel resistance and is independently associated with the risk of long-term recurrence of vascular events.

Objective:To modify Ganodermalucidum polysaccharides(GLP) with sulfate and observe the protective effect of Ganodermalucidum polysaccharide sulfate (GLPS) on the cerebral ischemia reperfusion injury in the rats,and to investigate its mechanism.Methods:GLP was modified by sulfation to obtain GLPS.A total of 100 SD rats were randomly divided into sham operation group, model group, GLP group (40 mg·kg-1·d-1), GLPS group (40 mg·kg-1·d-1) and nimodipine group (1 mg·kg-1·d-1).The cerebral ischemia reperfusion models were established by middle cerebral artery occlusion method in the rats.The neurologic deficit score and the content of water in brain tissue of the rats with cerebral ischemia reperfusion injury were detected and the activities of superoxide dismutase(SOD) and the levels of malondialdehyde (MDA) were detected.The levels of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB),tumor necrosis factor-alpha (TNF-α), interleukin-1 (IL-1) and interleukin-6 (IL-6) in the brain tissue homogenate were detected by ELISA.Western blotting method was used to detect the protein expression levels of HSP70 and p-Akt in the brain tissue of the rats.Results:Compared with model group, the neurological function scores of the rats in GLP group and GLPS group were decreased(P<0.01),the water contents in brain tissue were decreased(P<0.05), the SOD activities were increased and the MDA levels were decreased(P<0.05), and the levels of NF-κB, TNF-α, IL-1 and IL-6 were decreased(P<0.05);the effect in GLPS group was significantly better than that in GLP group(P<0.05).The results of Western blotting method showed that the p-Akt protein expression levels in the brain tissue of the rats in GLP and GLPS groups were increased compared with model group (P<0.05);compared with model group, the HSP-70 protein expression level in the brain tissue of the rats in GLPS group was increased(P<0.01),but the effect in GLP group was not obvious.Conclusion:Sulfation can significantly improve the protective effect of GLP on the cerebral ischemia reperfusion injury in the rats and its mechanism may be related to regulating the HSP70/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.

Objective To explore the clinical application value of blood routine examination in judgment of the activity of inflammatory bowel diseases.Methods 102 patients with inflammatory bowel disease were selected as the main research objects,60 healthy persons in the same period were selected as the control group,all of the subjects were received routine testing of blood,C-reactive protein and erythrocyte sedimentation rate,and the relationships between disease and blood routine indexes were compared.Results In UC group,the PLT,WBC RDW index levels of remission were higher than those of the control group,the differences were statistically significant(t =6.75,8.21,5.73,all P ＜0.05).In patients with UC,the HGB,HCTand MPV index levels were lower than those in the remission patients and the control group,the differences were statistically significant(t =8.12,11.03,6.72,9.35,10.23,8.71,all P ＜0.05).But the PLT index was higher than that of patients in remission,the RDW and WBC indexes were higher than those in the control group and the remission period,there were statistically significant differences (t =7.59,8.51,6.03,all P ＜ 0.05).The routine blood indexes of the relief patients of CD group had significant differences compared with the control group(t =6.17,7.29,9.01,10.35,8.27,7.69,8.01,9.91,allP ＜ 0.05),the HCT,MCV,HGB and MPV indexes were lower than those of the control group,the differences were statistically significant (t =9.01,10.35,8.27,9.9i,all P ＜ 0.05),the WBC,PLT,N and RDW indexes were higher than those of the control group,the differences were statistically significant (t =6.17,7.29,7.69,8.01,all P ＜ 0.05).In UC group,the HGB,HCT,MCV and MPV indexes were lower than those in the remission patients and the control group,there were statistically significant differences(t =8.19,6.97,7.53,9.02,all P ＜ 0.05).But the PLT,RDW,WBC and N indexes were higher than those in the remission patients and the control group,the differences were statistically significant (t =9.81,7.94,8.36,7.61,P ＜ 0.05).In the active patients of CD group,the WBC and N indexes were significantly higher than those of UC group,the differences were statistically significant (t =8.15,9.07,all P ＜ 0.05).In the remission patients of CD group,the PLT index was significandy higher than that in the remission patients of UC group,the difference was statistically significant (t =10.35,P ＜ 0.05).Conclusion Indexes of blood routine has obvious significance in activity of inflammatory bowel disease,and it is significant correlation with inflammation index,which can be used as one of the routine clinical method to determine the activity of inflammatory bowel diseases.