Bombesin receptor subtype-3 (BRS3) is an orphan G protein-coupled receptor (GPCR) that plays critical roles in energy homeostasis, glucose metabolism, and insulin secretion. Recent structural studies have elucidated BRS3 signaling mechanisms using synthetic ligands, including BA1 and MK-5046. However, the molecular basis of BRS3 activation by bioactive natural compounds and their derivatives, particularly those derived from traditional Chinese medicine, remains unclear. Here, we present high-resolution cryogenic electron microscopy (cryo-EM) structures of the human BRS3-G_q complex in both unliganded and active states bound by two herb-derived compounds (DSO-5a and oridonin), at resolutions of 2.9, 2.8, and 2.9 Å, respectively. These structures display distinct ligand recognition patterns between DSO-5a and oridonin. Although both compounds bind to the orthosteric pocket, they differentially engage the interaction network of BRS3, as demonstrated by mutagenesis studies assessing calcium mobilization and inositol phosphate 1 (IP1) accumulation. These findings enhance our understanding of BRS3 activation and provide valuable insights into the development of small-molecule BRS3 modulators with therapeutic potential.

Objective:To explore the value of 18F-fibroblast activation protein inhibitor (FAPI) PET/CT in the assessment of early-stage graft fibrosis (S1-S2) after liver transplantation (LT). Methods:From November 2021 to April 2022, 17 adult liver transplant recipients (12 males and 5 females; age (52.6±7.9) years) in the Affiliated Hospital of Qingdao University were enrolled retrospectively in this study. All 17 patients received laboratory examinations, FibroScan, 18F-FAPI PET/CT and liver biopsy. According to the Scheuer scoring system, hepatic tissue was divided into no fibrosis (S0) and early fibrosis (S1-S2). Independent-sample t test was used to compare SUV max between two groups, and Mann-Whitney U test was used to compare liver stiffness measurement (LSM). ROC curve analysis was used to evaluate the diagnostic efficacy of LSM and SUV max in the early fibrosis of liver grafts. Delong test was used to compare the difference of AUCs. Results:Among 17 adult LT recipients, 11 were in stage S0, 5 were in stage S1, and 1 was in stage S2. There were significant differences in LSM and SUV max between no fibrosis group and early fibrosis group (LSM: 5.4(4.7, 6.6) vs 12.9(5.6, 19.9) kPa, z=-2.01, P=0.044; SUV max: 1.7±0.8 vs 3.9±1.6, t=-3.14, P=0.019). The threshold value of LSM in predicting early-stage graft fibrosis was 8.2 kPa and the AUC was 0.80 (95% CI: 0.54-0.95), which was 2.0 and 0.92 (95% CI: 0.78-1.00) for SUV max respectively. There was no significant difference in AUC between the two tools ( z=0.80, P=0.421). Conclusion:18F-FAPI PET/CT can precisely evaluate the early fibrosis of allografts, with the similar diagnostic efficacy with FibroScan (LSM), which is expected to be a new non-invasive diagnostic tool for predicting the early-stage of graft liver fibrosis.

Objective To investigate the expression of miR-143-3p and ATG2B in serum of patients with bladder cancer and their relationship with clinicopathology,prognosis and survival of patients.Methods Eighty-eight patients who underwent radical bladder cancer surgery in our hospital from January 2017 to January 2019 were regarded as the research group,and 80 volunteers who underwent physical examination in our hospital were included as the control group.Quantitative real-time PCR(qRT-PCR)method was performed to detect the levels of miR-143-3p and ATG2B;The predictive value of miR-143-3p and ATG2B for bladder cancer was evaluated by ROC curve.Draw survival curve using Kaplan Meier method;The prognostic factors of bladder cancer patients were analyzed by Cox regression.Results The expression level of miR-143-3p in the serum of bladder cancer patients was lower than that in the control group(0.68±0.12 vs.0.87±0.26),and the expression level of ATG2B was higher(1.02±0.30 vs.0.61±0.18)(P＜0.05);the expression level of miR-143-3p in serum of postoperative patients was higher than that before operation(0.77±0.23 vs.0.68±0.12),and the expression level of ATG2B was lower than that before operation(0.85±0.26 vs.1.02±0.30)(P＜0.05);The study group showed a negative correlation between miR-143-3p and ATG2B expression(r=-0.454,r=-0.407,P＜0.01);The AUC of miR-143-3p in the diagnosis of bladder cancer was 0.846(95％CI:0.783-0.897),the sensitivity and specificity were 79.55％and 78.75％respectively;The AUC of ATG2B in the diagnosis of bladder cancer was 0.883(95％CI:0.824-0.927),the sensitivity and specificity were 78.41％and 81.25％respectively;The AUC for the diagnosis of bladder cancer was 0.939(95％CI:0.891-0.970),the sensitivity and specificity were 87.50％and 88.75％,respectively.The 3-year survival rate of the miR-143-3p low expression group was 34.88％(15/43)lower than that of the high expression group 77.78％(35/45),and the 3-year survival rate of the ATG2B high expression group was 40.91％(18/44)lower than that of the low expression group 72.73％(32/44).The differences were statistically significant(Log rank x2=18.055,8.658,P=0.000,0.003).Lymph node metastasis,distant metastasis,tumor pathological grade,TNM stage,miR-143-3p,ATG2B are all factors influencing the survival of bladder cancer patients.Conclusion The serum expression of miR-143-3p in patients with bladder cancer is low,and the expression of ATG2B is high,and the expressions of the two are closely related to the clinicopathological characteristics and prognosis of patients with bladder cancer.

Objective To investigate the role and underlying mechanism of atorvastatin on hyper-glycemia induced hemorrhagic transformation(HT)in a mouse model of cerebral ischemia.Meth-ods A total of 36 SPF-grade male C57BL/6 mice were randomly divided into sham operation group,HT model group and atorvastatin group,with 12 mice in each group.HE staining was used to observe cerebral hemorrhage,immunofluorescent staining was employed to detect the integrity of blood-brain barrier,and Western blotting was applied to measure the protein expression of IgG,ZO-1,occludin,claduin5,MMP-2 and-9 in ischemic penumbra brain tissues.Results Com-pared with sham operation group,the neurological deficit score,mortality rate,HT incidence,HT grading score,IgG fluorescence intensity,and protein levels of IgG,MMP-2 and-9 were signifi-cantly increased,while the protein levels of ZO-1,occludin and claudin5 were obviously decreased in the HT model group(P＜0.01).Atorvastatin treatment resulted in significantly lower neuro-logical deficit score(2.73±1.19 vs 3.91±0.94),mortality rate(16.7％vs 41.6％),HT incidence(58.3％vs 91.6％),HT grading score(1.00±1.04 vs 2.58±1.13),IgG fluorescence intensity(504.30±105.52 a.u vs 859.91±153.28 a.u),and protein levels of IgG(4.55±1.40 vs 12.06± 3.73),MMP-2(1.87±0.41 vs 2.95±0.68)and-9(1.47±0.24 vs 2.12±0.23)(P＜0.05,P＜0.01),and increased protein levels of ZO-1(1.55±0.20 vs 0.53±0.10),occludin(0.92±0.11 vs 0.35±0.07)and claudin5(0.58±0.04 vs 0.30±0.05)(P＜0.01)when compared with the HT model group.Conclusion Atorvastatin can reduce the permeability of blood-brain barrier by in-hibiting the activation of MMP-2 and MMP-9 and up-regulating the protein levels of ZO-1,occlu-din and claudin5,and thus attenuate hyperglycemia-induced HT.

Objective:To investigate the effect of mechano-growth factor（MGF）on osteoclast activity and its mechanism.Methods:The RAW264.7 precursor osteoclast cell line was cultured with 25 ng/ml macrophage-colony stimulating factor（M-CSF）and 30 ng/ml receptor activator of NF-κB ligand（RANKL），and identified by tartrate resistant acid phosphatase（TRAP）staining after 7 days of culture. Western blot anslysis was used to determine the effect of 45 ng/ml MGF on the phosphoinositide-3-kinase/protein kinase B（PI3K/AKT）signaling pathway in separated osteoclasts，including levels of AKT，phosphorylation（p）-AKT，lactation mammalian target of rapamycin（mTOR），p-mTOR and TRAP at 0，4，8 and 12 hours. Real-time fluorescence quantitative PCR was used to expressions of TRAP in osteoclasts at 0，4，8 and 12 hours. The PI3K/Akt phosphorylation inhibitor LY294002（20 μmol/L）combined with MGF（45 ng/ml）was used to act on osteoclasts，and expression levels of Akt，p-Akt，mTOR，p-mTOR and TRAP were detected by Western blot at 0，4，8 and 12 hours.Results:After culturing RAW264.7 cells with M-CSF and RANKL for 7 days，a large number of osteoclasts with positive TRAP staining can be obtained. Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-Akt and p-mTOR increased continuously from（2.18±0.34）pg/ml and（0.83±0.10）pg/ml at 0 hour to（3.86±0.36）pg/ml and（1.56±0.19）pg/ml at 12 hours（ P<0.05），and expression level of TRAP decreased significantly over time，from（5.66±0.47）pg/ml at 0 hour to（3.76±0.38）pg/ml at 12 hours（ P<0.05）. Real-time fluorescence quantitative PCR analysis of expression of TRAP in osteoclasts showed that MGF inhibited the expression of TRAP in osteoclasts，which decreased from 1.02±0.06 at 0 hour to 0.53±0.11 at 12 hours（ P<0.05）. After acting LY294002 combined with MGF on osteoclasts，Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-AKT and p-mTOR decreased significantly from（3.28±0.18）pg/ml and（3.29±0.22）pg/ml at 0 hour to（2.06±0.34）pg/ml and（2.04±0.20）pg/ml at 12 hours（ P<0.05），and expression level of TRAP had no significant difference over time（ P>0.05）. Conclusions:MGF inhibits osteoclast activity by inhibiting the expression of TRAP in osteoclasts through PI3K/Akt signaling pathway. LY294002 inhibits the expression of PI3K/Akt signaling pathway in osteoclasts，further verifying the mechanism of MGF inhibiting osteoclast activity，and this finding puts forward new ideas for clinical prevention and treatment of osteoporosis.

Objective To understand the prevalence,treatment and influence factors of hypertension in maintenance hemodialysis (MHD) patients in Anhui Province.Methods A total of 2724 adult patients on MHD from January 1st 2014 to March 31st 2014 in 26 hospitals of southern,northern and central Anhui Province were investigated.Their demographic characteristics,primary disease,complications,medications,dialysis and laboratory examination were explored.The prevalence treatment rate and control rate of hypertension were analyzed.Associated factors for controlling hypertension [systolic blood pressure (SBP) ＜ 140 mmHg and diastolic blood pressure (DBP) ＜ 90 mmHg] were assessed by logistic regression analysis.Results (1) The prevalence of hypertension in the hemodialysis patients was 87.0％.Their treatment rate and control rate were 93.2％ and 23.9％ respectively.The average of SBP was (145.90±21.18) mmHg,and the DBP on average was (83.60± 12.21) mmHg.The most commonly used anti-hypertensive drug is calcium channel blocker (88.2％).Over one third (45.7％) of patients were treated with two kinds of anti-hypertensive drug,26.2％ with 1 kind,21.7％ with 3 kinds,and 6.4％ with 4 kinds or more.(2) Compared with non-hypertension patients,patients with hypertension have older age,higher body mass index (BMI),phosphorus,SBP and DBP,as well as lower hemoglobin and Kt/V (all P ＜ 0.05).(3) The multivariate logistic regression analysis showed that Ca ＞ 2.50 mmol/L (OR=2.084,95％CI 1.008-4.307,P=0.047) positively correlated with controlling hypertension,while smoke (OR=0.594,95％CI 0.356-0.911,P=0.046) and BMI 18.5 ～ 23.9 kg/m2 (OR=0.516,95％CI 0.293-0.907,P=0.022) negatively correlated with it.Conclusions High prevalence yet low control rate of hypertension in MHD patients in Anhui Province were observed.Hypocalcemia may be a protective factor for hypertension control,while smoke and BMI may be risk factors for it.

[Objective]The purpose of this study is to provide a new idea for the prevention and treatment of PCOS in around-adolescence period through the cooperation of Wenshen Huatan prescription and food exchange. [Methods]By referring to and sorting out the related literatures, combined with clinical practice, this paper discusses how to form a specific plan of prevention and treatment of around -adolescence PCOS complicated with IGT by Wenshen Huatan prescription and food exchange. [Results]Food exchange can scientifically control diet, combined with kidney-warming and phlegm-Tiaojing, spleen phlegm, which has a remarkable effect on the treatment of around-adolescence PCOS complicated with IGT, such as improving clinical symptoms, shortening the menstrual cycle, improving endocrine disorders and insulin resistance.[Conclusion]It is significant for patients with IGT-type PCOS around-adolescence by the exchange of food with Wenshen Huatan prescription combined with diet intervention, which provides a new train of thought for clinical research.

Objective To prepare a 10-hydroxycamptothecin (10-HCPT) loaded folate-receptor targeted phase-change contrast agent (FR-HCPT-PNPCA),and to study the general characteristics including drug loading,phase changing and targeting capability in vitro.Methods Using a method of two-step emulsification,the phase-change nanoparticles loading anticancer drug (10-HCPT) with lipids shell and liquid pefluorocarbon core were prepared.The entrapment efficiency and the drug-loading amounts were studied by high performance liquid chromatography,and the phase transition of the nanoparticles after heating was observed.The targeting ability was evaluated on liver cancer cell line 7721 in vitro.Results The FR-HCPT-PNPCA,with a drug encapsulation rate of about 70.42 ％ and drug loading amounts of about 20.05 ％,was prepared successfully.When being heated to 70℃,obvious phase changing and microbubbles generating could be observed under microscope.In addition,a large amount of FR-HCPT-PNPCA particles could adhere specifically around the 7721 cells.Conclusion The prepared FR-HCPT-PNPCA,which has a stable characteristic and high performance of drug loading and tumor targeting,is expected to become a promising multifunctional molecular ultrasound probe for diagnosis and treatment of tumor.

Objective To investigate the expression of microRNA‐101 (miRNA‐101 ) in human gastric cancer ,and to explore its effects on proliferation ,migration and invasion of gastric cancer cell . Methods The expression of miRNA‐101 in 28 human gastric cancer tissues ,human gastric cell lines BGC‐823 , SGC‐7901 , MKN‐45 , AGS and human normal gastric epithelial cell line GES‐1 were determined by real time polymerase chain reaction (PCR) .Recombinant miRNA‐101 adenovirus vector was constructed . The effects of miRNA‐101 on gastric cancer proliferation was detected with cell proliferation assay .The ability of gastric cancer cell migration and invasion was assessed with Transwell assay .Gastric xenograft cancer model was established in BALB /c nude mice and the tumor size was compared .The t test was used for the statistical analysis .Results The expression of miRNA‐101 in gastric cancer tissues was 0 .661 ± 0 .396 ,which was lower than that of corresponding para carcinoma tissues (1 .128 ± 0 .697) ,and the difference was statistically significant (t = 10 .091 , P < 0 .01) .The expression of miRNA‐101 in normal gastric epithelial cell line GES‐1 was higher than those of gastric cancer cell lines BGC‐823 ,SGC‐7901 , MKN‐45 and AGS . There was significant suppression role of miRNA‐101 on MKN‐45 cells proliferation , and which also had inhibition role on cell migration and invasion of gastric cancer cell lines BGC‐823 ,SGC‐7901 ,MKN‐45 and AGS .At five weeks after MKN‐45 gastric xenograft cancer nude mice model established ,the tumor size of Ad‐miRNA‐101 group ((333 .56 ± 46 .71) mm3 ) was smaller than that of Ad‐enhanced green fluorescent protein (EGFP) group (806 .41 ± 51 .83) mm3 ,and the difference was statistically significant (t = 21 .431 , P < 0 .01 ) .Conclusion In gastric tissues and cells ,miRNA‐101 is a tumor suppressive miRNA and its downregulated expression involved in the genesis and development of gastric cancer ,which may be a new target of biological target therapy in gastric cancer .

Objective:To explore the relationship between thyroid function and cardiovascular diseases .Methods:A total of 980 patients undergoing thyroid function examination during hospitalization were selected . According to their thyroid function ,they were divided into normal thyroid function group (normal group ,n= 930) , hyperthy-roidism group (n=18) ,and hypothyroidism group (n=32) .Clinical data were analyzed ,blood lipids and coagula-tion function indexes were examined and compared among three groups . Results:Compared with normal group ,the incidence rate of atrial fibrillation significantly rose ,incidence rate of hypertension ,cardiac insufficiency signifi-cantly reduced;levels of TC、TG、 LDL-C、 HDL-C significantly reduced ,activated partial thromboplastin time significantly extended in hyperthyroidism group , P<0.01 all;incidence rate of coronary heart disease significantly rose ,levels of TG、HDL-C significantly rose ,levels of TC 、LDL-C significantly reduced in hypothyroidism group , P<0.01 all;Compared with hyperthyroidism group ,the incidence rate of hypertension ,coronary heart disease sig-nificantly rose ,atrial fibrillation significantly reduced ,P<0.05 or <0.01 ;levels of TG、LDL-C、HDL-C signifi-cantly rose ,TC level significantly reduced in hypothyroidism group , P<0. 01 all .Conclusion:Thyroid function is closely related to cardiovascular diseases .so it′s suggested that thyroid function detection should be regarded as a routine examination in patients with cardiovascular diseases and a follow-up index for those with thyroid dysfunc-tion .