Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

Objective:To evaluate the diagnostic value of coronary angiography-derived fractional flow reserve (FFR) and index of microcirculatory resistance (IMR) for identifying coronary functional abnormalities.Methods:This diagnostic study enrolled patients with clinically suspected or diagnosed coronary artery disease who underwent coronary angiography at Beijing Anzhen Hospital, TEDA International Cardiovascular Hospital, and Qilu Hospital of Shandong University between December 2021 and June 2022. All enrolled patients successfully underwent invasive wire-based FFR and IMR measurements during angiography. In a core laboratory, FFR and IMR for the target vessels were measured using artificial intelligence technology based on coronary angiographic images. Spearman correlation analysis was used to evaluate the correlation between angiography-derived FFR and wire-based FFR, and between angiography-derived IMR and wire-based IMR. Coronary hemodynamic abnormality was defined as FFR≤0.80; the diagnostic performance of angiography-derived FFR for identifying this abnormality was evaluated. Microcirculatory dysfunction was defined as IMR≥25; the diagnostic performance of angiography-derived IMR for identifying microcirculatory dysfunction was evaluated.Results:A total of 181 patients, aged (60.6±8.8) years, with 62 (34.3%) females, and 181 target vessels were included in the final analysis. Angiography-derived FFR showed a significant positive correlation with wire-based FFR ( r=0.78, P<0.001). For identifying coronary hemodynamic abnormality, angiography-derived FFR showed an accuracy of 89.0%, sensitivity of 88.8%, specificity of 89.1%, positive predictive value (PPV) of 88.8%, negative predictive value (NPV) of 89.1%, and an area under the receiver operating characteristic curve ( AUC) of 0.88. Angiography-derived IMR showed a significant positive correlation with wire-based IMR ( r=0.93, P<0.001). For identifying microcirculatory dysfunction, angiography-derived IMR demonstrated an accuracy of 89.5%, sensitivity of 86.8%, specificity of 90.2%, PPV of 70.2%, NPV of 96.3%, and an AUC of 0.95. Conclusion:Angiography-derived FFR and IMR exhibit strong correlations with their invasive wire-based counterparts and demonstrate high diagnostic value for assessing coronary hemodynamics and coronary microcirculatory function.

Objective:To observe the effects of Qizhi Tongluo Prescription on renal interstitial fibrosis in rats with membranous nephropathy based on the Shh/Gli1 signaling pathway; To explore its intervention mechanism.Methods:Totally 60 male SD rats were divided into blank group ( n=10) and model group ( n=50) using random number table method. The model of membranous nephropathy was established according to the modified Border method. The successfully modeling rats were divided into model group, benazepril group and Qizhi Tongluo Prescription low-, medium- and high-dosage groups using random number table method. Benazepril group was gavaged with benazepril hydrochloride 10 mg/kg, Qizhi Tongluo Prescription low-, medium- and high-dosage groups were gavaged with Qizhi Tongluo Prescription solution 1.22 g/kg, 2.43 g/kg and 4.86 g/kg, and blank group and model group were gavaged with equal volume of normal saline, once a day, for 4 weeks. The 24-hour urine was collected to detect the 24-hour urinary protein quantification, and the blood was taken from the abdominal aorta to detect the levels of total cholesterol (TC), triglyceride(TG), and serum albumin(ALB); the pathological changes of rat kidney were observed by light microscope and transmission electron microscope; the protein expressions of sonic hedgehog factor (Shh), zinc finger protein 1 (Gli1) and α-smooth muscle actin (SMA) in renal tissues were detected by immunohistochemistry; the protein expressions of Shh, Gli1, α-SMA, TGF-β1, Collagen Ⅳ and plasminogen activator inhibitor-1 (PAl-1) in renal tissues were detected by Western blot. Results:Compared with the model group, the quantitative level of 24-hour urinary protein of rats in each administration group decreased ( P<0.05), serum TC and TG levels increased ( P<0.05), ALB level decreased ( P<0.05), the positive expressions of Shh, Gli1, α-SMA protein in renal tissue decreased ( P<0.05), and the protein expressions of Shh, Gli1, α-SMA, Collagen Ⅳ, TGF-β1, PAI-1 in renal tissue decreased ( P<0.05). Conclusion:Qizhi Tongluo Prescription can improve renal interstitial fibrosis in membranous nephropathy rats, possibly by inhibiting the Shh/Gli1 signaling pathway to delay renal interstitial fibrosis.

Objective:To evaluate the diagnostic value of coronary angiography-derived fractional flow reserve (FFR) and index of microcirculatory resistance (IMR) for identifying coronary functional abnormalities.Methods:This diagnostic study enrolled patients with clinically suspected or diagnosed coronary artery disease who underwent coronary angiography at Beijing Anzhen Hospital, TEDA International Cardiovascular Hospital, and Qilu Hospital of Shandong University between December 2021 and June 2022. All enrolled patients successfully underwent invasive wire-based FFR and IMR measurements during angiography. In a core laboratory, FFR and IMR for the target vessels were measured using artificial intelligence technology based on coronary angiographic images. Spearman correlation analysis was used to evaluate the correlation between angiography-derived FFR and wire-based FFR, and between angiography-derived IMR and wire-based IMR. Coronary hemodynamic abnormality was defined as FFR≤0.80; the diagnostic performance of angiography-derived FFR for identifying this abnormality was evaluated. Microcirculatory dysfunction was defined as IMR≥25; the diagnostic performance of angiography-derived IMR for identifying microcirculatory dysfunction was evaluated.Results:A total of 181 patients, aged (60.6±8.8) years, with 62 (34.3%) females, and 181 target vessels were included in the final analysis. Angiography-derived FFR showed a significant positive correlation with wire-based FFR ( r=0.78, P<0.001). For identifying coronary hemodynamic abnormality, angiography-derived FFR showed an accuracy of 89.0%, sensitivity of 88.8%, specificity of 89.1%, positive predictive value (PPV) of 88.8%, negative predictive value (NPV) of 89.1%, and an area under the receiver operating characteristic curve ( AUC) of 0.88. Angiography-derived IMR showed a significant positive correlation with wire-based IMR ( r=0.93, P<0.001). For identifying microcirculatory dysfunction, angiography-derived IMR demonstrated an accuracy of 89.5%, sensitivity of 86.8%, specificity of 90.2%, PPV of 70.2%, NPV of 96.3%, and an AUC of 0.95. Conclusion:Angiography-derived FFR and IMR exhibit strong correlations with their invasive wire-based counterparts and demonstrate high diagnostic value for assessing coronary hemodynamics and coronary microcirculatory function.

Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

Humans ; Urinary Bladder Neoplasms/pathology* ; Carcinoma, Transitional Cell/pathology* ; Genetic Markers ; Biomarkers, Tumor/genetics* ; Urothelium/pathology*

Objective:To investigate the feasibility,safety and effectiveness of robot-assisted laparo-scopic buccal mucosa graft ureteroplasty in the treatment of complex long proximal ureteral stricture.Methods:The clinical data of 20 patients with proximal ureteral stricture undergoing robot-assisted lapa-roscopic buccal mucosa graft ureteroplasty admitted to the Department of Urology,Peking University First Hospital and Beijing Jiangong Hospital from July 2022 to January 2023 were prospectively collected and analyzed.Intraoperative conditions,postoperative complications and follow-up data were also recorded and analyzed.Results:The operations under robot-assisted laparoscopy were performed successfully in all the 20 patients without conversion to traditional laparoscopic surgery or open surgery.The study in-cluded 14 males and 6 females with a mean age of(41±11)years(range:19 to 60 years)and a mean body mass index of(24.3±3.6)kg/m2(range:18.2 to 31.8 kg/m2).There were 9 cases on the left side and 11 cases on the right side.The strictures of all the patients were located in the proximal segment of the ureter(including the ureteropelvic junction).The mean preoperative serum creatinine was(92.2±23.3)μmol/L(range:49.2 to 138.9 μmol/L),and the mean length of ureteral stricture was(2.8±0.9)cm(range:1.0 to 4.0 cm).Ten patients had previously undergone unsuccessful reconstructive surgery.During the operation,12 patients received posteriorly augmented anastomosis with ventral onlay.The mean length of the buccal mucosa graft harvested during the operation was(3.1±0.6)cm(range:2.0 to 4.3 cm),and the median width was 1.5 cm(range:1.0 to 2.0 cm).The omentum flap was used to wrap the reconstructed ureteral segment in all the 20 cases.The median operative time was 154 min(range:113 to 300 min),and the median estimated blood loss was 45 mL(range:0 to 100 mL).The median postoperative hospital stay was 4 d(range:4 to 14 d).The mean postoperative follow-up time was(15.0±1.7)months(range:12.5 to 17.9 months),and the surgical success rate was 100.0％in this study.After surgery,11 patients reported mild discomfort at the oral donor site,2 patients deve-loped urinary tract infection,and no postoperative complications were reported in the other 7 patients.The mean serum creatinine was(90.9±23.9)μmol/L(range:60.0 to 153.0 μmol/L)six months after surgery.Conclusion:Robot-assisted laparoscopic buccal mucosa graft ureteroplasty for the treatment of complex long proximal ureteral stricture has satisfactory efficacy without severe complications,which has shown good feasibility,safety and effectiveness.However,large sample studies and long-term follow-up are still needed to evaluate its long-term efficacy.

Objective:To summarize the clinical features, electroencephalogram (EEG) and magnetoencephalogram (MEG) of patients with pure paroxysmal kinesigenic dyskinesia (PKD) and PKD with epilepsy, so as to better distinguish them and guide the treatments.Methods:The clinical data of 200 patients diagnosed with PKD in the Outpatient Department of Xuanwu Hospital, Capital Medical University from 2000 to 2023 were analyzed retrospectively. The patients were divided into 2 groups: pure PKD (174 cases) and PKD with epilepsy (26 cases) according to whether accompanied by epilepsy. The differences in clinical features, drug therapy, EEG and MEG were compared between the 2 groups.Results:The clinical features of the 2 groups were essentially similar, and the proportion of PKD dyskinesia induced by emotional stress in the pure PKD group (54/174, 31.03%) was higher than that in the PKD with epilepsy group (2/26, 7.69%; χ 2=5.010, P=0.025). In terms of pharmacological response, carbamazepine was the most commonly used medication in both groups, but patients with PKD with epilepsy may need a higher therapeutic dosages (0.2-0.4 g/d, and gradually increased to 0.8 g/d) to effectively manage both dyskinesia and seizures. Regarding the EEG and MEG, the proportion of EEG abnormalities was higher in PKD patients with epilepsy, mainly manifested as focal spikes [1/70(1.43%) vs 9/21(42.86%), χ 2=24.268, P<0.001], together with aberrant MEG discharge (4/18 vs 3/5, χ 2=1.155, P=0.282). The MEG dipoles were mainly distributed in the brain regions close to the frontal lobe and central region. Conclusions:The clinical manifestations of motor symptoms of pure PKD and PKD with epilepsy are similar, and carbamazepine remains the most effective treatment. PKD patients with epilepsy have a higher proportion of abnormal EEG, mainly manifested as focal spikes, and are more likely to show abnormal discharge of MEG, which could be used to distinguish them.

OBJECTIVE: Diterpenoids with a wide variety of biological activities from Anoectochilus roxburghii, a precious medicinal plant, are important active components. However, due to the lack of genetic information on the metabolic process of diterpenoids in A. roxburghii, the genes involved in the molecular regulation mechanism of diterpenoid metabolism are still unclear. This study revealed the complex metabolic genes for diterpenoids biosynthesis in different organs of A. roxburghii by combining analysis of transcriptomics and metabolomics. METHODS: The differences in diterpenoid accumulation in roots, stems and leaves of A. roxburghii were analyzed by metabonomic analysis, and its metabolic gene information was obtained by transcriptome sequencing. Then, the molecular mechanism of differential diterpenoid accumulation in different organs of A. roxburghii was analyzed from the perspective of gene expression patterns. RESULTS: A total of 296 terpenoid metabolites were identified in the five terpenoid metabolic pathways in A. roxburghii. There were 38, 34, and 18 diterpenoids with different contents between roots and leaves, between leaves and stems, and between roots and stems, respectively. Twenty-nine metabolic enzyme genes with 883 unigenes in the diterpenoid synthesis process were identified, and the DXS and FDPS in the terpenoid backbone biosynthesis stage and CPA, GA20ox, GA3ox, GA2ox, and MAS in the diterpenoid biosynthesis stage were predicted to be the key metabolic enzymes for the accumulation of diterpenoids. In addition, 14 key transcription factor coding genes were predicted to be involved in the regulation of the diterpenoid biosynthesis. The expression of genes such as GA2ox, MAS, CPA, GA20ox and GA3ox might be activated by some of the 14 transcription factors. The transcription factor NTF-Y and PRE6 were predicted to be the most important transcription factors. CONCLUSION This study determined 29 metabolic enzyme genes and predicted 14 transcription factors involved in the molecular regulation mechanism of diterpenoid metabolism in A. roxburghii, which provided a reference for the further study of the molecular regulation mechanism of the accumulation of diterpenoids in different organs of A. roxburghii.

Objective:To compare the differences of reference intervals (RI) established by two types of indirect methods for 34 biochemical analytes, and to explore the possible factors that affect the consistency of the two methods.Methods:This was a retrospective study. Based on data of albumin (Alb), alkaline phosphatase (ALP), alanine aminotransferase (ALT), apolipoprotein A1(ApoA1), ApolipoproteinB (ApoB), aspartate aminotransferase (AST), calcium (Ca), cholinesterase (ChE), chloride (Cl), creatinine (Cr), high-sensitivity C-reactive protein (hsCRP), Cystatin (CysC), direct bilirubin (DBil), free fatty acid (FFA), glycated albumin(GA), gamma-glutamyltransferase (GGT), glucose (Glu), high density lipoprotein cholesterol (HDL-C), potassium (K), lactate dehydrogenase (LD), low density lipoprotein cholesterol (LDL-C), lipoprotein a [Lp (a)], sodium (Na), phosphorus (P), prealbumin (PA), superoxide dismutase (SOD), total bile acid (TBA), total bilirubin (TBil), total cholesterol (TC), total carbon dioxide (TCO 2), triglyceride (TG), total protein (TP), uric acid (UA) and urea (UR) of individuals who underwent physical examination at Peking Union Medical College Hospital from January 1, 2018 to December 31, 2019, Box-Cox algorithm was used to improve the data distribution and Tukey method was used to identify outliers. Variance component model was established, and standard deviation ratio (SDR) was calculated to determine whether the RIs of 34 biochemical analytes should be established according to age or sex The non-parametric method and kosmic algorithm were used to establish the RIs and 90% confidence intervals (CIs) of 34 biochemical analytes, and the coincidence of the 90% CIs of the reference limits for two methods was compared. Results:The skewness coefficients of ALP(male, female18-59), ALT, AST, hsCRP, DBil, GGT, Lp (a), TBA, TBil, TG, Glu, HDL-C(male) and CysC, GA, UR in the elderly group deviated from 0, and their kurtosis coefficients also deviated from 3. For these biochemical analytes, the point estimates of the RIs established by the two methods differed greatly and the 90% CIs did not overlap. The analytes with good normality were Alb, ApoA1, ApoB, Ca, ChE, Cl, Cr(E), CysC(18-59), FFA, GA(18-59), HDL-C(female), K, LDL-C, Na, P, PA, SOD, TC, TCO 2, TP and UR. The consistency is good. Except for Ca, 90% CIs of reference limits for some analyte between the two methods coincide with each other. Conclusions:The consistency of different indirect methods is affected by the normality of data.