ObjectiveBased on non-targeted metabolomics， to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus（T2DM） rats by Fuling Yunhua granules， and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats， half male and half female， were randomly divided into the control group， model group， and high， medium， low dose groups of Fuling Yunhua granules（20.70， 10.35， 5.18 g·kg^-1 in raw drug amount） and the positive drug group（pioglitazone hydrochloride tablets， 8.1 mg·kg^-1）. Except for the control group， other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin（STZ） to establish a T2DM rat model. After successful modeling， the treatment groups were administered the corresponding drugs by gavage， and the control group and model group were treated with an equal volume of saline by gavage， once/d， for 28 d. Fasting blood glucose（FBG） and glycosylated hemoglobin A1c（GHbA1c） levels were measured in all groups of rats during the administration period， and hematoxylin-eosin（HE） staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-LTQ-Orbitrap MS）， and the data were processed using principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Differential metabolites were identified by the Human Metabolome Database（HMDB） and the Kyoto Encyclopedia of Genes and Genomes（KEGG）， and screened for differential metabolites with variable importance in the projection（VIP） value>1， P<0.05， and fold change（FC）<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0， then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic（ROC） curves. ResultCompared with the control group， the FBG level of rats in the model group increased significantly（P<0.01）， the GHbA1c content tended to increase， but the difference was not statistically significant， and the pancreatic tissue of rats was obviously damaged， the number of pancreatic islets decreased， and the pancreatic β-cells were obviously reduced， atrophied and enlarged. Compared with the model group， the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration（P<0.05， P<0.01）， the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced（P<0.05）， and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid， D-amino acid metabolism， steroid hormone biosynthesis， and glycerophospholipid metabolism in rats. Compared with the model group， the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted， and the pathway enrichment analysis found that D-amino acid metabolism， retinol metabolism， glycine， serine and threonine metabolism， tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol， D-piperidinic acid， D-serine， and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.

BACKGROUND:Frog active peptides have rich activities,such as antibacterial and anti-tumor,and are expected to solve the problem of antibiotic resistance. OBJECTIVE:The active peptide QUB2984 was discovered in the skin secretions of Agalychnis callidryas.Its structure and properties were simulated by bioinformatics.The peptide was synthesized,purified,and identified and its biological functions were investigated. METHODS:Agalychnis callidryas skin secretions were collected by electrostimulation.The sequence of QUB2984 was obtained through constructing a cDNA library with isolated mRNA.BLAST was used for peptide sequence alignment.Besides that,Iterative Threading ASSEmbly Refinement(I-TASSER)and HeliQuest tools were used for protein secondary structure simulation.It was synthesized by solid-phase peptide synthesis,purified by reverse-phase high-performance liquid chromatography,and structurally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.The purified peptide was used to evaluate its biological activity.Its antibacterial effect was evaluated by the minimum inhibitory concentration method.Its cytotoxic effect was detected by MTT assay.Its safety was investigated by a hemolysis test. RESULTS AND CONCLUSION:(1)Peptide QUB2984 had basically α-spiral structure,with a relatively intact hydrophobic surface,and a certain destructive ability to biofilm.The third amino acid position of QUB2984 was composed of W and had a G-X-G structure.(2)The minimum inhibitory concentration of QUB2984 against gram-positive Staphylococcus aureus was 2 μmol/L,the minimum inhibitory concentration against gram-negative Escherichia coli was 2 μmol/L,and the minimum inhibitory concentration against the fungus Candida albicans was 8 μmol/L.(3)The active peptide QUB2984 had obvious inhibitory effect on human non-small cell lung cancer cells NCI-H838 at 10-5 mol/L concentration,and the hemolytic effect on horse red cells at 64 μmol/L concentration was 50%.(4)The results showed that QUB2984 had anti-bacterial and anti-cancer activity,and it had a positive charge of +3,which was conducive to contact with bacteria or cells.

Objective To explore the influencing factors of microcirculation dysfunction in patients with anterior wall acute myocardial infarction(AMI)and to establish a relevant prediction model.Methods A total of 130 patients with anterior wall AMI,whose microcirculation function was assessed by caIMR after receiving percutaneous coronary intervention(PCI)at Shanghai Sixth People's Hospital of China from January 2017 to September 2020,were enrolled in this study.The patients were divided into abnormal microcirculation resistance group(n=52)and normal microcirculation resistance group(n=78).The clinical data were compared between the two groups.The regression analysis was used to analyze the influencing factors of microcirculation dysfunction.Results In the abnormal microcirculation resistance group the contrast agent consumption,the onset-to-operation time,the Gensini total score and the LAD Gensini score were(121.92±31.37)mL,(10.51±5.12)min,(97.91±31.77)points and(69.36±13.15)points respectively,which were significantly higher than(109.03±28.2)mL,(4.94±2.94)min,(81.05±35.22)points and(54.45±23.48)points respectively in the normal microcirculation resistance group,the differences in the above indexes between the two groups were statistically significant(all P＜0.05).A prediction model covering interventional strategies was established,and its accuracy was higher than that of a conventional model,its AUC compared with the conventional model was 0.91 to 0.87,indicating that this model could well predict the risk of microcirculation dysfunction in patients with AMI after receiving PCI.Conclusion This prediction model can promptly identify high-risk microcirculation dysfunction patients with anterior wall AMI after receiving PCI.

Objective To investigate the effects of clopidogrel on behavioral,content of inflamma-tory cytokines,and microglial in mice with comorbidity of pain and depression.Methods Twenty-four male pathogen-free C57BL/6J mice,aged 8 weeks,weighing 23-27 g,were selected.The mice were randomly divided into three groups:sham group(group C),spared nerve injury(SNI)group(group S),and SNI+clopidogrel group(group L),8 mice in each group.The surgical procedure in group C was consistent with the SNI model,but the nerve and maintained nerve integrity were not damaged.Group S was prepared with the SNI model,and group L was given clopidogrel 10 mg/kg since 21 days after the SNI model was prepared for 14 days continuously.Mechanical withdrawal threshold(MWT)was detected 1 day before surgery,7,14,21,28,and 35 days after surgery.The tail suspension test(TST)and forced swim test(FST)were performed 35 and 37 days after surgery,and the motionless time was calculated.Then the mice were killed,and the right hippocampal tissue was harvested to detect the concentration of IL-1β,IL-6,and TNF-α by enzyme-linked immunosorbent assay(ELISA),the expression of IL-1β,IL-6,and TNF-α mRNA by reverse transcription-polymerase chain reaction(RT-PCR),and the number of microglia by immunofluores-cence staining.Results Compared with group C,MWT were significantly decreased 7,14,21,28,and 35 days after operation,the immobility time of TST and FST were significantly prolonged,the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α were significantly increased,and the number of micro-glia were significantly increased in groups S and L(P＜0.05).Compared with group S,MWT were signifi-cantly increased 28 and 35 days after operation,the immobility time of TST and FST were significantly short-ened,the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α were significantly decreased,and the number of microglia was significantly reduced in group L(P＜0.05).Conclusion Clopidogrel can reduce the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α,inhibit the inflammatory reaction,reduce the number of activated microglia,and improve the and chronic neuropathic pain and de-pression in mice with comorbidity of pain and depression.

Objective To investigate the effects of denosumab(DEN)and alendronate(AL)on bone mineral density(BMD)around the prosthesis after total hip arthroplasty(THA).Methods A retrospective analysis was conducted on the clinical data of primary osteoporosis(POP)patients who underwent THA at the Affiliated Jinhua Hospital of Zhejiang University School of Medicine from March 2022 to March 2023,the patients were divided into the DEN group and the AL group according to the postoperative protocols.The main observation index was the bone density around the implant(Gruen zone:1st and 7th zones;time points:postoperative day 5(T1),6 months(T2),and 12 months(T3);detection method:dual-energy X-ray absorptiometry).The secondary observation index was the bone metabolism index(bone calcitonin(OC),β-collagen special sequence(β-Crosslaps),and type 1 procollagen N-terminal peptide(P1NP);time points:preoperative(T4),6 months(T2),and 12 months(T3);detection method:enzyme-linked immunosorbent assay).Results A total of 120 patients were included in the study,with 59 in the DEN group and 61 in the AL group.At baseline,there was no statistically significant difference in serum OC,β-Crosslaps and P1NP levels between the two groups(P＞0.05).Periprosthetic BMD showed a decreasing trend in both groups,and periprosthetic BMD was higher in the DEN group than in the AL group in both Gruen zone 1 and Gruen zone 7 at the T2 and T3 time points(P＜0.05).In terms of bone transformation markers,serum OC,β-Crosslaps and P1NP levels showed decreasing trends with prolonged treatment time in both groups(P＜0.05).In addition,serum OC,β-Crosslaps and P1NP levels were significantly lower in the DEN group than in the AL group at T2 and T3 time points(P＜0.05).Conclusion Compared with AL,DEN is more effective in preventing periprosthetic bone loss of the proximal femur and has a better safety profile.

【Objective】 To investigate the feasibility of laparoscopic simple prostatectomy (LSP) in the treatment of large volume benign prostate hyperplasia (BPH). 【Methods】 Clinical and follow-up data of 30 patients with large volume BPH treated with LSP in our hospital during Feb.2019 and Dec.2021 were retrospectively analyzed. All patients underwent extraperitoneal LSP operation. The perioperative and 1-12 month postoperative follow-up data were analyzed. 【Results】 The average prostate volume was (92.4±38.9) mL, operation time (125±45) min, and weight of resected prostate (60.25±16.90) g. The hemoglobin decreased by (12.21±7.25) g/d after operation. No blood transfusion was needed. There was no need for bladder irrigation after operation in 21 cases (70%), and 9 cases (30%) had bladder irrigation time of (0.95±0.49) d. The postoperative catheter indwelling time was (6.92±2.51) d, and hospital stay (5.36±1.63) d. During the follow-up of (9.25±5.4) months, there was 1 case of postoperative intestinal obstruction (Clavien-Dindo grade II), 1 case of transient urinary incontinence (Clavien-Dindo grade I), and 1 case of delayed hematuria (Clavien-Dindo grade I). No urethral stricture occurred. The maximum urinary flow rate (Qmax), post-void residual urine volume (PVR), International Prostate Symptom Score (IPSS) and quality of life (QoL) 3 months after operation were significantly improved compared with those before operation (P<0.05). There was no significant difference in sexual function before and after operation (P>0.05). 【Conclusion】 LSP is safe and effective in the treatment of large volume BPH. It has advantages of complete resection of glands, minor bleeding and short postoperative bladder irrigation time. However, it still needs to be confirmed by a prospective control study of large samples.

Objective : To investigate the effects of aflatoxin B1 (AFB1) on the biophysical properties and cytoskeleton structure of human hepatocellular carcinoma cells (HCCs) . Methods: HepG2 cells were respectively treated with 0 , 0. 01 , 0. 1 , 1 , 5 , 10 μmol/L AFB1 for 24 h and 48 h , and the cell viability was measured by CCK⁃8 kit.Based on this result , the influences of 10 μmol/L AFB1 on the osmotic fragility , membrane fluidity , electrophoretic mobility (EPM) and F ⁃actin structure of cells were analyzed. Subsequently , total RNAs were extracted and the PCR. Results: The increased viability of HepG2 cells was induced by AFB1 in a dose⁃dependent manner after 48h treatment. After treated with 10 μmol/L AFB1 , the anti⁃hypotonic ability and EPM of HepG2 cells were en⁃hanced. The content of F ⁃actin in HepG2 cells increased obviously , while the mRNA expression levels of the main cytoskeleton binding proteins were altered. Conclusion AFB1 can affect the biophysical properties , cytoskeleton structure and its binding proteins of HepG2 cells , which may be directly related to its toxic action.

Objective:To identify the etiology and genetics of the human parainfluenza virus type 3 (HPIV3) virus which caused an acute respiratory tract infection outbreak in a primary school in Shenyang.Methods:Throat swab samples were collected from 17 students of the primary school where the epidemic of acute respiratory infection outbreak in December 2020 in Shenyang, Liaoning province. TaqMan low-density arrays (TLDA) real-time PCR was performed to simultaneously detect multiple respiratory pathogens. The HN gene was amplified using nested RT-PCR and sequenced, followed by phylogenetic analysis for those HPIV3 positive samples.Results:Of the 17 specimens, 10 were HPIV3 positive by TLDA Real-time PCR, and were accompanied by conditional pathogen infection, consequently, amplification result ed in 7 complete HN sequences. Phylogenetic analysis showed that the infected HPIV3 virus of the outbreak belonged to HPIV subtype C3a. All the 7 strains detected in this study belonged to subbranch C3a.1 evolutionary branch, with a nucleotide homology of 99.9%, a nucleotide homology of 94.56 with the prototype strain Wash/47885/57 and 99.5% with the most phylogenetically close strain of ZJ/11-s-165/KP690785/CHN/11.Conclusions:The HPIV3 virus caused the acute respiratory tract infection outbreak in Shenyang in 2020 and HPIV subtype C3a1 was detected firstly in Northeast China.

Objective:To analyze the risk of missed diagnosis in patients with PI-RADS score>3 and negative prostate initial biopsy and to explore its risk factors.Methods:The clinical data of 268 patients with negative prostate biopsy in Northern Jiangsu People's Hospital from May 2013 to December 2018 were retrospectively analyzed. The patients were divided into observation group (PI-RADS score>3) and control group (PI-RADS score≤ 3) according to different PI-RADS scores. There were insignificant differences in age [(67.4(60.0, 74.0)years and 65.6(66.5, 72.0)years], prostate volume of initial biopsy [62.4(40.0, 72.0)ml and 60.8(38.0, 77.0)ml], biopsy cores [ 20.6(18.0, 22.0)cores and 20.4(18.0, 22.0)cores] between the observation group (n=124) and the control group(n=144)(all P>0.05). But there were significant differences in PSA [17.5(6.5, 23.0)ng/ml and 11.5(6.3, 12.0)ng/ml], PSAD[0.316(0.128, 0.363)ng/ml 2 and 0.211(0.106, 0.256)ng/ml 2], prostate inflammation of the initial biopsy [70 (56.5%) and 32 (22.2%)] between the observation group and the control group(all P<0.05). According to the follow-up results after the initial biopsy, the two groups of repeated biopsy were compared.Furthermore, Logistic regression was used to conduct univariate and multivariate analysis to explore the risk factors of patients with PI-RADS>3 for positive repeated biopsy. At the same time, the receiver operating characteristic curve (ROC curve) was used to analyze the accuracy of the risk factors. Results:There were significant differences in repeated biopsy rate [ 27.4%(34/124)and 14.6%(21/144)], CsPCa detection rate[ 41.4%(14/34) and 4.8%(1/21)]between the observation group and the control group(all P<0.05). The positive rate of repeated biopsy in the observation group (41.1%) was higher than that in the control group (23.8%), but there was no statistical difference ( P=0.248). The risk of positive repeated biopsies in the observation group was 2.24 times than that in the control group. Univariate analysis found repeated biopsy PSA ( P =0.02, OR=1.438, 95% CI 1.161-1.896), PSA ratio (repeated biopsy PSA/initial biopsy PSA) ( P=0.011, OR=10.087, 95% CI 1.714-59.36) were risk factors for positive of repeated biopsy in patients with PI-RADS score >3. Multivariate analysis also found that repeated biopsy PSA ( P=0.017, OR=1.15, 95% CI 1.076-2.123), PSA ratio ( P=0.032, OR=10.2, 95% CI 0.883-116.168) were risk factors for positive repeated biopsy. ROC curve analysis, the accuracy of repeated biopsy PSA (AUC=0.971, P<0.001, 95% CI 0.926-1.000), PSA ratio (AUC=0.839, P=0.001, 95% CI0.707-0.971) to predict positive of repeated biopsy were high. The cut-off values were 21.3 ng/ml and 1.4, respectively. The accuracy was higher when combines repeated biopsy PSA with PSA ratio (AUC=0.993, P<0.001, 95% CI 0.974-1.000). Conclusions:Patients with negative PI-RADS score > 3 have a higher risk of missed diagnosis of CsPCa than those with PI-RADS score≤3. When PSA>21.3 ng/ml and PSA ratio>1.4 during follow-up, the possibility of missed diagnosis in the initial biopsy is high.

Objective: To evaluate the effect of multimodal analgesia using periprostatic nerve block anesthesia (PNB) combined with flurbiprofen in patients undergoing transperineal template-guided prostate biopsy (TTPB). Methods: Totally 166 patients (aged (68.2±9.1) years, range: 47 to 81 years) who received TTPB from October 2017 to June 2018 at Department of Urology, Northern Jiangsu People′s Hospital Affiliated to Yangzhou University were enrolled prospectively. All the patients were randomly divided into 2 groups. The observation group (n=79) was given flurbiprofen axetil 1 mg/kg intravenously for half an hour before operation and lidocaine was used for PNB before the biopsy. The control group (n=87) was given normal saline combined with PNB. A visual analog scale (VAS) and visual numeric scale (VNS) were used to assess the patients′ pain and quantify their satisfaction at two time points: VAS-1 and VNS-1: during biopsy procedure, VAS-2 and VNS-2: 30 min after the procedure. The date were compared by t test, χ² test, Fisher exact test and two-way repeated measures anova analysis between the 2 groups. Results: The age, total prostate volume, serum prostate-specific antigen and the number of cores were comparable among the 2 groups (P>0.05). The VAS-1 scores of the control group and the observation group were 2.8±1.7, 1.9±1.2, respectively, and the VNS-1 were 3.1±0.7, 3.4±0.3, respectively. The VAS-1 were significantly lower in observation group than in control group (F=3.904, P=0.000). Conversely, the VNS-1 were higher in observation group (F=3.526, P=0.000). At 30-minute postoperative, the VAS-2 and VNS-2 were 0.7±0.4 and 3.7±0.2 in the control group, respectively. The VAS-2 and VNS-2 were 0.6±0.5 and 3.8±0.1 in the observation group, respectively. There were no significant differences in the pain scores or the satisfaction scores between the 2 groups (F=1.429, 2.825; P=0.136, 0.083). The incidence of overall complications was 26.4% (23/87) in the control group and 25.3% (20/79) in the observation group, with no statistical difference between the 2 groups (χ²=0.027, P=0.869). And the complications had no statistically significant difference among the 2 groups including hematuria, urinary retention, infection, hematospermia, vascular and neurological reactions, nausea, vomiting, dizziness, headache, and respiratory depression (P>0.05). Conclusion The multimodal analgesia induced by PNB and flurbiprofen could effectively relieve the pain for patients who received TTPB.