Objective To investigate the effects of Danlong Xingnao Prescription on learning and memory ability and microglia activation in rats with vascular dementia(VD)based on HMGB1/RAGE/NF-κB pathway.Methods Ten rats were randomly selected from 72 rats as a sham-operation group.The remaining rats were treated with modified bilateral common carotid artery ligation method to prepare the VD model.The 50 successful model rats were randomly divided into model group,nimodipine group(10.8 mg/kg)and Danlong Xingnao Prescription low-,medium-and high-dosage groups(3.7,7.4,14.8 g/kg),with 10 rats in each group.The administration groups were given relevant solution for gavage,the sham-operation group and model group were given the same amount of normal saline for consecutive 28 d.Morris water maze test was performed to evaluate learning and memory abilities of rats,the morphology in the hippocampus were observed by HE staining,the contents of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α in hippocampal tissue were detect by ELISA,RT-PCR was used to detect high mobility group protein B1(HMGB1),receptor of advanced glycation end product(RAGE),nuclear factor(NF)-κB p65 and regulatory RNase-1(Regnase-1)mRNA expression in hippocampal tissue,immunohistochemistry and Western blot were used to detect the protein expressions of ion calcium binding adapter molecule 1(Iba1),HMGB1,RAGE,NF-κB p65 and Regnase-1 in hippocampal tissue.Results Compared with the sham-operation group,the escape latency of rats was prolonged,and the number of crossings through the original platform was increased in the model group(P<0.01),the pyramidal cells in the hippocampus were reduced and irregularly shaped,with unclear cell and nuclear membranes,and a significant number of necrotic neurons were visible,the contents of IL-1β,IL-6 and TNF-α in hippocampal tissue increased(P<0.01),the mRNA expressions of HMGB1,RAGE and NF-κB p65 in hippocampal tissue increased(P<0.01),while the mRNA expression of Regnase-1 decreased(P<0.01),the protein expressions of Iba1,HMGB1,RAGE and NF-κB p65 increased(P<0.01),while the protein expression of Regnase-1 decreased(P<0.01).Compared with the model group,the escape latency of rats was shortened in Danlong Xingnao Prescription groups,the number of crossings through the original platform was reduced(P<0.05,P<0.01),the neuronal structure of hippocampal tissue was significantly improved,the number of necrotic neurons was reduced,and the contents of IL-1β,IL-6 and TNF-α in hippocampal tissue reduced(P<0.05,P<0.01),the mRNA expressions of HMGB1,RAGE and NF-κB p65 in hippocampal tissue decreased,the mRNA expression of Regnase-1 increased(P<0.05,P<0.01),the protein expression of Iba1,HMGB1,RAGE and NF-κB p65 decreased,the protein expression of Regnase-1 increased(P<0.05,P<0.01).Conclusion Danlong Xingnao Prescription can improve the learning and memory ability of VD rats,and its mechanism may be related to inhibiting the activation of HMGB1/RAGE/NF-κB pathway and increasing Regnase-1 expression,thereby inhibiting the activation of microglia.

Objective:To observe the effects of electroacupuncture(EA)on learning and memory ability and nucle-ar factor E2 related factor 2(Nrf2),glutathione peroxidase 4(GPX4),NAD(P)H-quinone oxidoreductase 1(NQO1),reactive oxygen species(ROS)in hippocampus of neonatal rats with hypoxic-ischemic brain damage(HIBI)and its anti-oxidative stress mechanism.Methods:A total of 76 neonatal rats were randomly divided into Sham opera-tion group(Sham),HIBI model group(HIBI),electroacupuncture group(HIBI+EA),and Nrf2 inhibitor ML385 group(HIBI+EA+ML385).The neonatal rat model of HIBI was established by classical Rice method.The HIBI+EA group was treated with electroacupuncture for 30 min/d for 14 consecutive days.In the HIBI+EA+ML385 group,30 mg/kg ML385 was intraperitoneally injected at 1 h before each electroacupuncture intervention.Morris water maze test was performed on 21 days after modeling to test the learning and memory ability of neonatal rats in each group.Nissl staining and HE staining were used to observe the morphology of neurons in the hippocampal CA1 region,and DHE flu-orescent probe was used to detect the expression of ROS in the hippocampal CA1 region.The contents of Nrf2,GPX4 and NQO1 in hippocampus were detected by Western blot.Results:Compared with the Sham group,the HIBI group had severe pathological damage,a prolonged escape latency,a decreased number of platform crossings,a significantly increased expression of ROS,and a significantly decreased expression of Nrf2,GPX4,and NQO1(P<0.05).Com-pared with HIBI group,the pathological damage was significantly attenuated,the escape latency was shortened,the number of platform crossings was increased,the expression of ROS was decreased,and the expression of Nrf2,GPX4 and NQO1 was increased in HIBI+EA group(P<0.05).Compared with HIBI+EA group,the pathological damage was aggravated,the escape latency was prolonged,the number of crossing the platform was decreased,the expression of ROS was increased,and the expression of Nrf2,GPX4,and NQO1 was decreased in HIBI+EA+ML385 group(P<0.05).Conclusion:Electroacupuncture can effectively improve the learning and memory ability of hypoxic-ischemic neonatal rats,which is related to its effect in reducing oxidative stress in hippocampal neurons by regulating Nrf2 signa-ling pathway.

Objective To investigate the effects of Danlong Xingnao Prescription on learning and memory ability and microglia activation in rats with vascular dementia(VD)based on HMGB1/RAGE/NF-κB pathway.Methods Ten rats were randomly selected from 72 rats as a sham-operation group.The remaining rats were treated with modified bilateral common carotid artery ligation method to prepare the VD model.The 50 successful model rats were randomly divided into model group,nimodipine group(10.8 mg/kg)and Danlong Xingnao Prescription low-,medium-and high-dosage groups(3.7,7.4,14.8 g/kg),with 10 rats in each group.The administration groups were given relevant solution for gavage,the sham-operation group and model group were given the same amount of normal saline for consecutive 28 d.Morris water maze test was performed to evaluate learning and memory abilities of rats,the morphology in the hippocampus were observed by HE staining,the contents of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α in hippocampal tissue were detect by ELISA,RT-PCR was used to detect high mobility group protein B1(HMGB1),receptor of advanced glycation end product(RAGE),nuclear factor(NF)-κB p65 and regulatory RNase-1(Regnase-1)mRNA expression in hippocampal tissue,immunohistochemistry and Western blot were used to detect the protein expressions of ion calcium binding adapter molecule 1(Iba1),HMGB1,RAGE,NF-κB p65 and Regnase-1 in hippocampal tissue.Results Compared with the sham-operation group,the escape latency of rats was prolonged,and the number of crossings through the original platform was increased in the model group(P<0.01),the pyramidal cells in the hippocampus were reduced and irregularly shaped,with unclear cell and nuclear membranes,and a significant number of necrotic neurons were visible,the contents of IL-1β,IL-6 and TNF-α in hippocampal tissue increased(P<0.01),the mRNA expressions of HMGB1,RAGE and NF-κB p65 in hippocampal tissue increased(P<0.01),while the mRNA expression of Regnase-1 decreased(P<0.01),the protein expressions of Iba1,HMGB1,RAGE and NF-κB p65 increased(P<0.01),while the protein expression of Regnase-1 decreased(P<0.01).Compared with the model group,the escape latency of rats was shortened in Danlong Xingnao Prescription groups,the number of crossings through the original platform was reduced(P<0.05,P<0.01),the neuronal structure of hippocampal tissue was significantly improved,the number of necrotic neurons was reduced,and the contents of IL-1β,IL-6 and TNF-α in hippocampal tissue reduced(P<0.05,P<0.01),the mRNA expressions of HMGB1,RAGE and NF-κB p65 in hippocampal tissue decreased,the mRNA expression of Regnase-1 increased(P<0.05,P<0.01),the protein expression of Iba1,HMGB1,RAGE and NF-κB p65 decreased,the protein expression of Regnase-1 increased(P<0.05,P<0.01).Conclusion Danlong Xingnao Prescription can improve the learning and memory ability of VD rats,and its mechanism may be related to inhibiting the activation of HMGB1/RAGE/NF-κB pathway and increasing Regnase-1 expression,thereby inhibiting the activation of microglia.

Objective To establish an analytical method for the simultaneous determination of 18 perfluoroalkyl compounds(PFCs)in tea leaves using a quick,easy,cheap,effective,rugged,and safe(QuEChERS)method for sample pretreatment combined with ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Methods The target analytes—18 PFCs—included 13 carboxylic acid PFCs(perfluorobutanoic acid[PFBA],perfluoropentanoic acid[PFPeA],perfluorohexanoic acid[PFHxA],perfluoroheptanoic acid[PFHpA],perfluorooctanoic acid[PFOA],perfluorononanoic acid[PFNA],perfluorodecanoic acid[PFDA],perfluoroundecanoic acid[PFUdA],perfluorododecanoic acid[PFTrDA],perfluorotridecanoic acid[PFTeDA],perfluorotetradecanoic acid[PFHxDA],perfluorohexadecanoic acid[PFHpS],and perfluorooctadecanoic acid[PFODA])and 5 sulfonic acid PFCs(perfluorobutanesulfonic acid[PFBS],perfluorohexanesulfonic acid[PFHxS],perfluoroheptanesulfonic acid[PFHpS],perfluorooctanesulfonic acid[PFOS],and perfluorodecanesulfonic acid[PFDS]).The QuEChERS pretreatment parameters were systematically optimized using the response surface methodology.The tea leave samples were extracted with an 80%acetonitrile solution and subsequently purified by adding a mixed absorbent consisting of 20 mg N-propyl-ethylenediamine(PSA),210 mg graphitized carbon black GCB),and 60 mg octadecylsilane(C18).The supernatant was concentrated by nitrogen blowing and subsequently re-dissolved in 50%methanol-2 mmol/L ammonium acetate solution.The re-dissolved solution was injected into the UHPLC-MS/MS for analysis.The target analytes were separated on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm,1.7 μm).The mobile phases consisted of methanol(phase A)and 2 mmol/L aqueous ammonium acetate(phase B),with a gradient elution procedure.The total running time was 18 min.The mass spectrometry analysis was conducted using an electrospray ionization source in negative ionization mode and multi-reaction monitoring(MRM),with quantification performed using the internal standard curve method.The greenness of the analytical method was assessed using Analytical GREEnness calculator(AGREE)and the Analytical Eco-Scale method(AES).Results Under the optimized conditions,the limits of detection(LODs)and limits of quantification(LOQs)of the method were 0.005 7-1.23 ng/g and 0.019-4.09 ng/g,respectively.The average recoveries of most target compounds were 71.1%-117.9%,with relative standard deviations(RSDs)below 15%.The AGREE index of the method was 0.49,and the AES score was 76.At least one PFC was detected in each of the 132 tea leave samples,and the detection rate of carboxylic acid PFC was higher than that of sulfonic acid PFC.The highest detection rates were observed for PFBA at 97.74%,PFHpA at 93.23%,and PFOA at 92.24%.In contrast,PFHpS,PFUdA,PFDoA,PFHxDA,and PFODA were not detected in the samples.Conclusion The proposed method has the advantages of simplicity,rapidity and sensitivity,and is suitable for the analysis of PFCs in tea leaves.The method has high greenness with minimal impact on the operator and the environment.The widespread presence of PFC contamination in tea leaves available in the market warrants strengthened monitoring and regulatory control.

Objective:To observe the effects of electroacupuncture(EA)on learning and memory ability and nucle-ar factor E2 related factor 2(Nrf2),glutathione peroxidase 4(GPX4),NAD(P)H-quinone oxidoreductase 1(NQO1),reactive oxygen species(ROS)in hippocampus of neonatal rats with hypoxic-ischemic brain damage(HIBI)and its anti-oxidative stress mechanism.Methods:A total of 76 neonatal rats were randomly divided into Sham opera-tion group(Sham),HIBI model group(HIBI),electroacupuncture group(HIBI+EA),and Nrf2 inhibitor ML385 group(HIBI+EA+ML385).The neonatal rat model of HIBI was established by classical Rice method.The HIBI+EA group was treated with electroacupuncture for 30 min/d for 14 consecutive days.In the HIBI+EA+ML385 group,30 mg/kg ML385 was intraperitoneally injected at 1 h before each electroacupuncture intervention.Morris water maze test was performed on 21 days after modeling to test the learning and memory ability of neonatal rats in each group.Nissl staining and HE staining were used to observe the morphology of neurons in the hippocampal CA1 region,and DHE flu-orescent probe was used to detect the expression of ROS in the hippocampal CA1 region.The contents of Nrf2,GPX4 and NQO1 in hippocampus were detected by Western blot.Results:Compared with the Sham group,the HIBI group had severe pathological damage,a prolonged escape latency,a decreased number of platform crossings,a significantly increased expression of ROS,and a significantly decreased expression of Nrf2,GPX4,and NQO1(P<0.05).Com-pared with HIBI group,the pathological damage was significantly attenuated,the escape latency was shortened,the number of platform crossings was increased,the expression of ROS was decreased,and the expression of Nrf2,GPX4 and NQO1 was increased in HIBI+EA group(P<0.05).Compared with HIBI+EA group,the pathological damage was aggravated,the escape latency was prolonged,the number of crossing the platform was decreased,the expression of ROS was increased,and the expression of Nrf2,GPX4,and NQO1 was decreased in HIBI+EA+ML385 group(P<0.05).Conclusion:Electroacupuncture can effectively improve the learning and memory ability of hypoxic-ischemic neonatal rats,which is related to its effect in reducing oxidative stress in hippocampal neurons by regulating Nrf2 signa-ling pathway.

Standardized reporting is a crucial factor affecting the use of patient guidelines （PGs）， particularly in the reporting and presentation of recommendations. This paper introduced the current status of PG reporting， including the research on PG content and presentation formats， and provided comprehensive recommendations for PG reporting from aspects such as overall framework， recommendations， presentation format， and readability. First， the presentation of PG recommendations should include clearly defined clinical questions， recommendations and their rationale， and guidance on how patients should implement the interventions； for specific content in the PG， such as level of evidence， level of recommendation， it is recommended to explain in text the reasons for giving different levels of recommendation， i.e.， to present the logic behind giving the level of recommendation to the patient； additional information needed in the recommendation framework should be supplemented by tracing references or authoritative textbooks and literature that support the recommendations. Subsequently， the PG text should be written based on the Reporting Checklist for Public Versions of Guidelines （RIGHT-PVG） reporting framework. Finally， to enhance readability and comprehension， it is recommended to refer to the Patient Education Materials Assessment Tool （PEMAT） for translating PG content. To enhance the readability of PGs， it is suggested to present the PG content in a persona-lized and layered manner.

Since the concept of patient versions of guidelines （PVGs） was introduced into China， several PVGs have been published in China， but we found that there is a big difference between the concept of PVG at home and abroad， and the reason for this difference has not been reasonably explained， which has led to ambiguity and even misapplication of the PVG concept by guideline developers. By analyzing the background and purpose of PVGs， and the understanding of the PVG concept by domestic scholars， we proposed the term patient guidelines （PGs）. This refers to guidelines developed under the principles of evidence-based medicine， centered on health issues that concern patients， and based on the best available evidence， intended for patient use. Except for the general attribute of providing information or education， which is typical of common health education materials， PGs also provide recommendations and assist in decision-making， so PGs include both the patient versions of guidelines （PVG） as defined by the Guidelines International Network （GIN） and "patient-directed guidelines"， i.e. clinical practice guidelines resulting from the adaptation or reformulation of recommendations through clinical practice guidelines.

At present， the process and methodology of patient guidelines （PGs） development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers， we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus， so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage， the construction stage， and the completion stage in general， but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network， i.e.， team formation， screening of recommendations， guideline drafing， user testing and feedback， approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process （forming a team， determining the scope of the PG， searching， evaluating and integrating evidence， forming recommendations， writing the guideline， and expert review）. Based on the different models， we sort out the process and methods of PG development and introduce the specific methods of PG development， including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines， with a view to providing reference for guideline developers and related researchers.

Objective To study the changes of serum metabolites in the patients with chronic kidney disease osteoporosis(CKD-OP)to provide the new biomarkers for the early diagnosis of CKD-OP.Methods A total of 22 patients with definitely diagnosed CKD visiting in this hospital from April to Novem-ber 2023 were selected as the study subjects,and 22 subjects with physical examination in this hospital at the same period were included for conducting the control study.With the lumbar vertebral T value ≤-2.5 as the standard,the subjects were divided into the CKD-OP group(n=11),CKD non-OP group(CKD-NOP group,n=11),simple osteoporosis group(OP group,n=11)and healthy control group(NC group,n=11).The liq-uid chromatography-mass spectrometry(LC-MS)coupling technique was used to analyze the differences in se-rum metabolites among the four groups,the potential biomarkers of CKD-OP was screened,and the correla-tion between the potential biomarkers with lumbar vertebra bone mineral density(BMD),serum bone-derived alkaline phosphatase(BALP)and serum tartrate-resistant acid phosphatase-5b(TRACP-5b)was studied.Re-sults With the receiver's operating characteristic(ROC)curve's area under curve(AUC)＞0.9 as the con-dition,and four potential biomarkers of CKD-OP were screened,which were phosphorylcholine,lysophosphati-dylcholine(18∶2/0∶0),capric acid,and allantoin respectively.Serum phosphorylcholine was positively corre-lated with lumbar vertebra BMD(r=0.601,P＜0.05)and negatively correlated with serum BALP and TRACP-5b(r=-0.729,-0.623,P＜0.05).Serum allantoin was positively correlated with lumbar vertebra BMD(r=0.483,P＜0.05)and negatively correlated with serum BALB(r=-0.494,P＜0.05).Serum lyso-phosphatidylcholine(18∶2/0∶0)was positively correlated with lumbar vertebra BMD(r=0.640,P＜0.05),and negatively correlated with serum BALP and TRACP-5b(r=-0.628,-0.548,P＜0.05).Serum capric acid was negatively correlated with lumbar vertebra BMD(r=-0.444,P＜0.05)and positively corre-lated with serum BALB(r=0.587,P＜0.05).Conclusion The screened four endogenous potential biomarkers provide the new research ideas for the early diagnosis and treatment efficacy monitoring of CKD-OP.

Objective To investigate the influence of spectral CT monochromatic imaging technique on image quality and liver vol-ume measurement in computed tomography portal venography(CTPV).Methods A total of 120 patients who underwent contrast-enhanced abdominal CT examination were prospectively selected and randomly divided into group A and group B.The group A(n=60)was scanned with conventional parameters such as 120 kVp,Smart mA mode,and image reconstruction of 60％adaptive statistical iterative reconstruction-Veo(ASIR-V);while the group B(n=60)was with instantaneous switching 80/140 kVp,gemstone spectral imaging(GSI)Assist mode,and image reconstruction of 60％ASIR-V.In group B,six subgroups of images from 75 to 50 keV(with 5 keV interval)were recorded as subgroups B1 to B6.On the axial images,the CT values and standard deviation(SD)values of por-tal vein,liver and erector spinae were measured,and then the contrast-to-noise ratio(CNR)of portal vein and liver were calculated.The Liver Segmentation software was used to segment the liver in groups A and B,the liver volumes by automatic segmentation and manual segmentation(golden standard)were recorded,and then the volume difference rate was calculated.The overall image quality and automatic liver segmentation results were evaluated by two radiologists using a 5-point scale.Results In terms of overall image quality,subgroup B6 achieved the highest score and was superior to group A(P＜0.001).In terms of liver segmentation,subgroup B3 had the highest score and was superior to group A(P＜0.001).With the decrease of keV,the CT values,SD values and CNR of portal vein and liver in group B were gradually increased(P＜0.05),in which subgroup B6 was higher than that in group A(P＜0.001).The volume difference rate initially decreased and then increased with the decrease of keV.Except for subgroups B2 and B3,the differences were statistically significant between other subgroups and group A(P＜O.001),and the subgroup B3 had the lowest volume difference rate.Conclusion Spectral CT monochromatic ima-ging technique has an influence on CTPV image quality and liver volume measurement.The 50 keV images are the best for displaying portal vein,and the automatic liver segmentation volume of 65 keV images is closest to the real liver volume.