Objective To introduce a modified technique of right ventricular outflow tract (RVOT) reconstruction using a handmade bicuspid pulmonary valve crafted from expanded polytetrafluoroethylene (ePTFE) and to summarize the early single-center experience. Methods Patients with complex congenital heart diseases (CHD) who underwent RVOT reconstruction with a handmade ePTFE bicuspid pulmonary valve due to pulmonary regurgitation at Guangdong Provincial People’s Hospital from April 2021 to February 2022 were selected. Postoperative artificial valve function and right heart function indicators were evaluated. Results A total of 17 patients were included, comprising 10 males and 7 females, with a mean age of (18.18±12.14) years and a mean body weight of (40.94±19.45) kg. Sixteen patients underwent reconstruction with a handmade valved conduit, with conduit sizes ranging from 18 to 24 mm. No patients required mechanical circulatory support, and no in-hospital deaths occurred. During a mean follow-up period of 12.89 months, only one patient developed valve dysfunction, and no related complications or adverse events were observed. The degree of pulmonary regurgitation was significantly improved post-RVOT reconstruction and during follow-up compared to preoperative levels (P<0.001). Postoperative right atrial diameter, right ventricular diameter, and tricuspid regurgitation area were all significantly reduced compared to preoperative values (P<0.05). Conclusion The use of a 0.1 mm ePTFE handmade bicuspid pulmonary valve for RVOT reconstruction in complex CHD is a feasible, effective, and safe technique.

Neuropathic pain is a debilitating pathological condition that presents significant therapeutic challenges in clinical practice. Unfortunately, current pharmacological treatments for neuropathic pain lack clinical efficacy and often lead to harmful adverse reactions. As G protein-coupled receptors (GPCRs) are widely distributed throughout the body, including the pain transmission pathway and descending inhibition pathway, the development of novel neuropathic pain treatments based on GPCRs allosteric modulation theory is gaining momentum. Extensive research has shown that allosteric modulators targeting GPCRs on the pain pathway can effectively alleviate symptoms of neuropathic pain while reducing or eliminating adverse effects. This review aims to provide a comprehensive summary of the progress made in GPCRs allosteric modulators in the treatment of neuropathic pain, and discuss the potential benefits and adverse factors of this treatment. We will also concentrate on the development of biased agonists of GPCRs, and based on important examples of biased agonist development in recent years, we will describe universal strategies for designing structure-based biased agonists. It is foreseeable that, with the continuous improvement of GPCRs allosteric modulation and biased agonist theory, effective GPCRs allosteric drugs will eventually be available for the treatment of neuropathic pain with acceptable safety.

Pyran- and furanocoumarins are key representatives of tetrahydropyrans and tetrahydrofurans, respectively, exhibiting diverse physiological and medical bioactivities. However, the biosynthetic mechanisms for their core structures remain poorly understood. Here we combined multiomics analyses of biosynthetic enzymes in Peucedanum praeruptorum and in vitro functional verification and identified two types of key enzymes critical for pyran and furan ring biosynthesis in plants. These included three distinct P. praeruptorum prenyltransferases (PpPT1-3) responsible for the prenylation of the simple coumarin skeleton 7 into linear or angular precursors, and two novel CYP450 cyclases (PpDC and PpOC) crucial for the cyclization of the linear/angular precursors into either tetrahydropyran or tetrahydrofuran scaffolds. Biochemical analyses of cyclases indicated that acid/base-assisted epoxide ring opening contributed to the enzyme-catalyzed tetrahydropyran and tetrahydrofuran ring refactoring. The possible acid/base-assisted catalytic mechanisms of the identified cyclases were theoretically investigated and assessed using site-specific mutagenesis. We identified two possible acidic amino acids Glu303 in PpDC and Asp301 in PpOC as vital in the catalytic process. This study provides new enzymatic tools in the epoxide formation/epoxide-opening mediated cascade reaction and exemplifies how plants become chemically diverse in terms of enzyme function and catalytic process.

BACKGROUND:Previous studies have confirmed that the new compound cottonrose hibiscus leaf gel plaster has a good effect in the treatment of acute soft tissue swelling. OBJECTIVE:To observe the clinical efficacy of compound cottonrose hibiscus leaf gel plaster in the treatment of synovitis of the knee joint. METHODS:Seventy-two patients with knee synovitis were selected from Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from December 2019 to May 2021.These patients were randomly divided into a trial group and a control group,with 36 cases in each group.The trial group was treated with compound cottonrose hibiscus leaf gel plaster,once a day,12 hours each time,while the control group was treated with Diclofenac Diethylamine Emulgel,twice a day.After 28 days of treatment,visual analog scale score,WOMAC Osteoarthritis Index score,quality of life score(SF-36),thickness of knee synovium and comprehensive curative effect were compared between the two groups. RESULTS AND CONCLUSION:(1)Visual analog scale scores after treatment were lower than those before treatment(P<0.05).Visual analog scale scores in the trial group after 7,14 and 28 days of treatment were lower than those in the control group(P<0.05).The WOMAC Osteoarthritis Index scores of the two groups after treatment were lower than those before treatment(P<0.05),and the WOMAC Osteoarthritis Index scores in the trial group after 7,14 and 28 days of treatment were lower than those in the control group(P<0.05).(3)The SF-36 quality of life score in the two groups after 28 days of treatment was higher than that before treatment(P<0.05).SF-36 quality of life score in the trial group after 28 days of treatment was higher than that in the control group(P<0.05).(4)After 28 days of treatment,the thickness of knee synovium in the trial group was less than that in the control group(P<0.05),and the effective rate in the trial group was higher than that in the control group(P<0.05).(5)These findings indicate that compared with Diclofenac Diethylamine Emulgel,the compound cottonrose hibiscus leaf gel plaster can better relieve knee pain,enhance knee joint function,reduce synovial hyperplasia,and elevate the overall quality of life of patients.

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.

Objective To analyze the current status and frontier trends of research on the application of cognitive-motor dual task training(CMDT)in stroke. Methods Relevant literatures on the application of CMDT in stroke were retrieved from CNKI,Wanfang data,VIP,SinoMed and Web of Science Core Collection from inception to October 11,2023,and was analyzed with CiteSpace 6.2R4. Results A total of 285 articles were included with 124 in Chinese and 161 in English.The annual number of publications showed a general upward trend.United States,Canada,Netherlands,China and United Kingdom were the lead-ing countries in terms of output in English.The scholar with the most publications in Chinese was Zheng Jiejiao,and the Department of Rehabilitation Medicine in Huadong Hospital of Fudan University was the leading institu-tion for Chinese publications.Vrije University Amsterdam was the leading institution for English publications.The most frequent Chinese keywords were gait,falls,balance,cognition and postural control.The most frequent English keywords were dual-task,walking,gait,balance and cognitive-motor interference.Bursting keywords from the past two years included gait training,cognitive tasks,balance ability and cognitive-motor interference. Conclusion The researches on the application of CMDT in stroke are on the rise,with hotspots including gait training,cognitive tasks and cognitive-motor interference.The mechanisms of CMDT and the development of optimal CMDT rehabilitation protocols for stroke may be researched more in the future.

Objective:To investigate the effect of Chinese medicine He's Yangchao recipe on premature ovarian insufficiency(POI)and its relationship with mitochondrial function of ovarian granulose cells in an animal model.Methods:Thirty-six female C57BL/6J mice were randomly divided into blank control group,model group,low-,medium-and high-dose He's Yangchao recipe treatment group and coenzyme Q10(Q10)treatment group(positive control).The POI model was induced by a single intraperitoneal injection of cyclophosphamide(90 mg/kg).The animals were sacrificed after 21 days.Primary granulose cells were obtained from POI mice and treated with He's Yangchao recipe,ERβ inhibitor PHTPP,and He's Yangchao recipe+PHTPP in vitro for 24 h,respectively.Ovarian histopathological changes were observed by hematoxylin-eosin(HE)staining,ATP levels were detected by luciferase assay,mtDNA copy numbers were detected by quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR),mitochondrial structure changes were observed by transmission electron microscopy,protein and mRNA expression levels of estrogen receptor β(ERβ),peroxisome proliferator-activated receptor γ coactivator 1α(PGC1α),mitochondrial transcription factor A(TFAM),and superoxide dismutase 2(SOD2)were detected by Western blotting and qRT-PCR.Results:The ovarian tissue in model group exhibited few secondary and tertiary follicles,whereas the He's Yangchao recipe groups and Q10 group had abundant secondary and tertiary follicles.Compared with the blank control group,ATP and mtDNA levels in model group decreased(P＜0.01),mitochondrial crista disappeared or abnormal vacuolated structure increased;the protein and mRNA levels of ERβ,PGC1α,TFAM,and SOD2 decreased(all P＜0.01).ATP production increased in granulose cells of high-dose He's Yangchao recipe group and Q10 group;mtDNA copy numbers increased(P＜0.05 or P＜0.01);abnormal mitochondrial structure was reduced;the protein and mRNA expressions of ERβ,PGC1α,TFAM,and SOD2 increased(P＜0.05 or P＜0.01).Compared with the PHTPP intervention group,the proportion of normal mitochondrial structure in the granulose cells of He's Yangchao recipe+PHTPP group was higher;ATP content increased(P＜0.05 or P＜0.01);mtDNA copy numbers increased(P＜0.05 or P＜0.01);the protein and mRNA expression of ERβ,PGC1α,TFAM and SOD2 increased(P＜0.05 or P＜0.01).Conclusions:He's Yangchao recipe can regulate mitochondrial biogenesis through ERβ/PGC1α/TFAM pathway to improve ovarian function in POI mice.

Objective:To investigate the effect of methylene blue (MB) on motor dysfunction and its mechanism in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) mouse models.Methods:Forty healthy male C57BL/6 mice were randomly divided into 4 groups: control group, model group, low-dose treatment group and medium-dose treatment group ( n=10); PD mouse models were established by intraperitoneal injection of 25 mg/kg/d MPTP for a consecutive 7 d; low-dose treatment group and medium-dose treatment group were pretreated intraperitoneally with MB 2 mg/kg/d or MB 10 mg/kg/d for a consecutive 3 d, respectively; and then, MPTP 25 mg/kg/d+MB 2 mg/kg/d or MPTP 25 mg/kg/d+MB 10 mg/kg/d were injected intraperitoneally into the low-dose treatment group or medium-dose treatment group for a consecutive 7 d (MPTP and MB were given at 12 h of interval). Eight d after modeling, open field experiment, pole climbing experiment and rod rotating experiment were carried out to evaluate the spontaneous movement, coordination, endurance and motor ability. And then, the mice were sacrificed; immunofluorescent staining was used to observe tyrosine hydroxylase (TH) expression in the substantia nigra; Western blotting was used to detect the expressions of TH, α-synuclein, nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-Caspase-1 and Gasdermin D (GSDMD) in the striatum and substantia nigra of mice. Contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-18 in the substantia nigra and striatum of mice were detected by ELISA. Results:Compared with the control group, the model group had shortened residence time in rod rotating, prolonged descent time in rod climbing, reduced total movement distance in open field, decreased number of TH-positive cells in the substania nigra, decreased TH protein levels in the substania nigra and striatum, and increased NLRP3, ASC, cleaved-Caspase-1, GSDMD and GSDMD-N protein levels in the substania nigra and striatum, and increased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). Compared with the model group, low-dose treatment group and medium-dose treatment group had prolonged residence time in rod rotating, shortened descent time in rod climbing, increased total movement distance in open field, increased number of TH-positive cells in the substania nigra, and increased TH protein levels in the substania nigra and striatum, decreased NLRP3, ASC, and cleaved-Caspase-1 levels in the substania nigra and striatum, and decreased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). No statistical differences in the above indexes were noted between the low-dose treatment group and medium-dose treatment group ( P>0.05). Conclusion:Low-/medium-dose MB can ameliorate motor dysfunction in PD mouse models, whose mechanism may be related to downregulate NLRP3 inflammasome and inhibit neuroinflammatory response to reduce dopaminergic neuron pyroptosis.

OBJECTIVE To explore the effects of the water extract of Morinda officinalis on the expressions of sex hormones and receptors in bisphenol A （BPA）-contaminated mice. METHODS Totally 60 male Kunming mice were randomly divided into control group， model group， and low-dose， medium-dose and high-dose groups of M. officinalis water extract （20， 40， 60 mg/kg）， with 12 mice in each group. The model group and M. officinalis water extract groups were given BPA intragastrically [50 mg/（kg·d）， once a day， for 4 consecutive weeks] to establish the BPA-contamination model of mice. After modeling， each drug group was gavaged with the corresponding drug solution， once a day， for 4 consecutive weeks. After the last medication， the body weight and testicular weight of the mice in each group were weighed， the histopathological changes in the testis were observed， and the serum sex hormones [luteinizing hormone （LH）， follicle-stimulating hormone （FSH）] contents and the mRNA and protein expressions of LH receptor （LHR） and FSH receptor （FSHR） in the testicular tissues were detected. RESULTS Compared with the control group， the testicular tissues of mice in the model group had structural degeneration， loose connections between spermatogenic cells and Sertoli cells， obvious lacunae and reduced number of spermatogenic cells； the mRNA and protein expressions of LHR and FSHR in testicular tissues were significantly down-regulated （P＜0.01）， but there were no significant changes in their body weights， testicular weights， and serum contents of LH and FSH （P＞0.05）. Compared with the model group， the histopathological changes of testicular tissues of mice in each dose group of M. officinalis water extract were improved to different degrees， and the mRNA and protein expressions of LHR and FSHR in testicular tissues were up-regulated to different degrees （P＜0.05 or P＜ 0.01）， and some indicator levels were similar to those of the control group （P＞0.05）. However， there were no significant changes in their body weights， testicular weights， and serum contents of LH and FSH （P＞0.05）. CONCLUSIONS The water extract of M. officinalis has a certain improvement effect on testicular injury in BPA-contaminated mice， which might be related to its increase in the mRNA and protein expressions of LHR and FSHR.

Objective:To investigate the predictive value of myocardial perfusion-related parameters measured by echocardiography on the prognosis of patients with coronary artery disease at 90 d after percutaneous coronary intervention (PCI).Methods:Eighty-five patients with coronary artery disease who underwent PCI in Linping Branch of the Second Affiliated Hospital of Zhejiang University from October 2020 to October 2022 were selected retrospectively. Patients were divided into the occurrence myocardial perfusion injury group (40 cases) and the non-occurrence myocardial perfusion injury group (45 cases). The quantitative echocardiographic parameters of left ventricular end-diastolic internal diameter (LVEDD) and left ventricular end-systolic internal diameter (LVESD) were compared between the two groups, and the effects of LVEDD and LVESD on the risk of myocardial perfusion injury after PCI in patients with coronary artery disease were analyzed. Logistic regression was used to analyze the factors influencing the major adverse cardiovascular events (MACE) of patients with coronary artery disease at 90 d after PCI. A line graph model was constructed, the receiver operating characteristic (ROC) curve and decision curve analysis (DCA) were constructed to verify the predictive efficiency of the line graph model.Results:Compared with the day of admission, the levels of LVEDD and LVESD were increased at 2 and 6 h after PCI, and showed an increasing trend at each time point ( P<0.05). The levels of LVEDD and LVESD at 2 and 6 h after PCI in the occurrence myocardial perfusion injury group were higher than those in the non-occurrence myocardial perfusion injury group: (45.56 ± 2.35) mm vs. (43.27 ± 2.12 ) mm, (47.87 ± 3.56) mm vs. (45.73 ± 2.98) mm; (33.49 ± 2.32) mm vs. (31.29 ± 2.29) mm, (35.62 ± 3.03) mm vs. (33.74 ± 2.12) mm, there were statistical differences ( P<0.05). The risk of myocardial perfusion injury was 4.469 and 6.081 times higher in patients with high levels of LVEDD and LVESD than in patients with low levels. The results of Logistic regression analysis showed that coronary multiple lesions, time from onset to balloon dilation, coronary Gensini score and LVEDD, LVESD were independent influencing factors of MACE at 90 d after PCI in patients with coronary artery disease ( P<0.05). Based on the above five independent influencing factors, a column line graph model was established to predict the risk of poor prognosis at 90 d after PCI. According to the column line graph model, the coronary multiple lesions were scored as 5; the corresponding prognostic adverse risk increased with the increase of time from onset to balloon dilation, coronary Gensini score and quantitative echocardiographic parameters LVEDD and LVESD; the internal validation of the column line graph prediction model was performed, and the C-index of the model was 0.978; the calibration curve showed that the model had good differentiation and accuracy in predicting the risk of MACE at 90 d after PCI. The results of ROC curve showed, the area under the curve (AUC) was 0.955 (95% CI 0.952 - 1.000). The DCA showed that when the line graph model was in the high risk threshold range (0 - 0.9), the prediction of the model had clinical practical value and the net benefit of patients was high. Conclusions:The abnormal increase of LVEDD and LVESD determined by echocardiography is associated with myocardial perfusion injury in patients with coronary heart disease after PCI, and a line graph model based on LVEDD and LVESD can predict the occurrence of MACE at 90 d after PCI, so as to guide early risk assessment and prevention.