1.Development and preliminary application of a one-step RT-qPCR method for de-tection of porcine epidemic diarrhea virus based on immunomagnetic beads
Shunli YANG ; Xiaojuan LYU ; Li LI ; Xiaoqing ZHANG ; Yupeng FANG ; Tian ZHAO ; Jiqiao XIA ; Jie ZHANG ; Zhixin FU ; Yongsheng LIU
Chinese Journal of Veterinary Science 2025;45(9):1817-1823
In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.
2.Development and preliminary application of a one-step RT-qPCR method for de-tection of porcine epidemic diarrhea virus based on immunomagnetic beads
Shunli YANG ; Xiaojuan LYU ; Li LI ; Xiaoqing ZHANG ; Yupeng FANG ; Tian ZHAO ; Jiqiao XIA ; Jie ZHANG ; Zhixin FU ; Yongsheng LIU
Chinese Journal of Veterinary Science 2025;45(9):1817-1823
In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.
3.Application of lidocaine in enhanced recovery after radical prostatectomy
Yupeng LYU ; Pei WANG ; Lihong HU
China Modern Doctor 2024;62(19):93-96,99
Objective To investigate the effect of intravenous lidocaine on enhanced recovery of elderly men after laparoscopic radical prostatectomy.Methods A total of 78 patients who planned to undergo laparoscopic radical prostatectomy in Affiliated Lihuili Hospital of Ningbo University from September to December 2023 were selected as study objects,and they were divided into control group and experimental group according to random number table method,with 39 patients in each group.Experimental group received intravenous lidocaine of 1.5mg/kg during induction of anesthesia,followed by micropump injection at a rate of 2mg/(kg·h),and control group was injected with an equal amount of normal saline.The intraoperative dosage of propofol and remifentanil was recorded.Visual analogue scale(VAS)score,postoperative postanesthesia care unit(PACU)residence time,first anal exhaust time,first defecation time,first getting out of bed activity time,length of hospital stay,number of effective patient-controlled intravenous analgesia(PCIA)compressions within 48h after surgery,number and dosage of sufentanil addition,quality of recovery-40(QoR-40)scale score,and occurrence of adverse reactions within 48h after surgery were compared between two groups.Results The intraoperative dosage of propofol and remifentanil,the number of effective PCIA compression,the number of sufentanil addition and the amount of sufentanil in postoperative 48h in experimental group were significantly lower than those in control group(P<0.05).The VAS scores of experimental group at 2h,6h,12h,24h and 48h were significantly lower than those of control group(P<0.05).The PACU residence time,first anal exhaust time,first defecation time,first getting out of bed activity time and hospital stay time of experimental group were significantly shorter than those of control group(P<0.05).The incidence of nausea and vomiting in experimental group was significantly lower than that in control group within postoperative 48h(P<0.05),and no adverse reactions such as drowsiness,respiratory depression,and severe bradycardia occurred in both groups.48h after surgery,the comfort,emotion,pain scores and total scores of QoR-40 in experimental group were significantly higher than those in control group(P<0.05).Conclusion In elderly men undergoing laparoscopic radical prostatectomy,intravenous infusion of lidocaine can reduce the amount of perioperative anesthetic drugs,reduce postoperative pain and adverse reactions,and promote postoperative recovery.
4.Application of ultrasound guided percutaneous portal vein puncture indocyanine green positive staining in laparoscopic anaphylectomy of S8 segments of liver
Yupeng TANG ; Yao HUANG ; Jianxing ZENG ; Jiahui LYU ; Guangwen LIU ; Qian LIN ; Xiaoling YU ; Yongyi ZENG
Chinese Journal of Hepatobiliary Surgery 2024;30(12):881-885
Objective:To investigate the value of ultrasound guided preoperative percutaneous portal vein puncture indocyanine green (ICG) in laparoscopic anatomic liver S8 resection.Methods:The clinical data of 15 patients with hepatocellular carcinoma who underwent laparoscopic anabolic S8 hepatectomy in Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2023 to December 2023 were retrospectively analyzed, including 13 males and 2 females with the age of (57.77±11.53) years. Clinical data such as China liver cancer staging (CNLC), target portal vein puncture operation time, operation time, intraoperative blood loss, postoperative complications and perioperative death were recorded.Results:Among the 15 patients, 11 were CNLC Ia stage and 4 were CNLC Ib stage. The operation time of target portal vein puncture for the 15 patients was (11.33±4.51) min, and they all successfully completed percutaneous portal vein puncture ICG fluorescence injection under ultrasound guidance. However, the dyeing effect was not up to expectations for 2 patients. All patients completed laparoscopic anatomical resection of the liver S8 segment, including 6 cases of resection of the liver S8 segment, 8 cases of resection of the posterior segment of the liver S8 segment, and 1 case of resection of the anterior segment of the liver S8 segment. The operative time of laparoscopic anabolic S8 hepatectomy was 181.3 (119.0, 237.0) min, the intraoperative blood loss was 203.3 (100.0, 300.0) ml, there was no intraoperative blood transfusion, and the postoperative hospital stay was (12.00±3.88) d. No positive surgical margin was found in postoperative pathology, and the tumor diameter of 15 patients was (3.29±1.10) cm. No complications of Clavien-Dindo grade Ⅲ or above occurred after surgery, and no perioperative death occurred, and only 1 patient developed pulmonary infection after surgery.Conclusion:Preoperative ultrasound-guided percutaneous portal vein puncture ICG can effectively assist laparoscopic anatomic S8 hepatectomy in patients with hepatocellular carcinoma.
5.Application of ultrasound guided percutaneous portal vein puncture indocyanine green positive staining in laparoscopic anaphylectomy of S8 segments of liver
Yupeng TANG ; Yao HUANG ; Jianxing ZENG ; Jiahui LYU ; Guangwen LIU ; Qian LIN ; Xiaoling YU ; Yongyi ZENG
Chinese Journal of Hepatobiliary Surgery 2024;30(12):881-885
Objective:To investigate the value of ultrasound guided preoperative percutaneous portal vein puncture indocyanine green (ICG) in laparoscopic anatomic liver S8 resection.Methods:The clinical data of 15 patients with hepatocellular carcinoma who underwent laparoscopic anabolic S8 hepatectomy in Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2023 to December 2023 were retrospectively analyzed, including 13 males and 2 females with the age of (57.77±11.53) years. Clinical data such as China liver cancer staging (CNLC), target portal vein puncture operation time, operation time, intraoperative blood loss, postoperative complications and perioperative death were recorded.Results:Among the 15 patients, 11 were CNLC Ia stage and 4 were CNLC Ib stage. The operation time of target portal vein puncture for the 15 patients was (11.33±4.51) min, and they all successfully completed percutaneous portal vein puncture ICG fluorescence injection under ultrasound guidance. However, the dyeing effect was not up to expectations for 2 patients. All patients completed laparoscopic anatomical resection of the liver S8 segment, including 6 cases of resection of the liver S8 segment, 8 cases of resection of the posterior segment of the liver S8 segment, and 1 case of resection of the anterior segment of the liver S8 segment. The operative time of laparoscopic anabolic S8 hepatectomy was 181.3 (119.0, 237.0) min, the intraoperative blood loss was 203.3 (100.0, 300.0) ml, there was no intraoperative blood transfusion, and the postoperative hospital stay was (12.00±3.88) d. No positive surgical margin was found in postoperative pathology, and the tumor diameter of 15 patients was (3.29±1.10) cm. No complications of Clavien-Dindo grade Ⅲ or above occurred after surgery, and no perioperative death occurred, and only 1 patient developed pulmonary infection after surgery.Conclusion:Preoperative ultrasound-guided percutaneous portal vein puncture ICG can effectively assist laparoscopic anatomic S8 hepatectomy in patients with hepatocellular carcinoma.
7.A case of postoperative obstructive hydrocephalus after pinealoma surgery onset with depressive symptoms
Meiti WANG ; Chongze WANG ; Ni ZHOU ; Yupeng CHEN ; Yinghua TAO ; Fang WANG ; Qinyu LYU ; Wu HONG
Chinese Journal of Psychiatry 2023;56(6):453-456
Pinealoma often comorbid with obstructive hydrocephalus due to their specific location and type, therefore, patients with pinealoma need to be treated by a combined ventriculoperitoneal shunt (VPS) along with surgical resection. Shunt failure is the most common complication after surgery. In this paper, we report a case of obstructive hydrocephalus caused by shunt obstruction after pineal gland tumor resection combined with ventriculoperitoneal shunt. The patient first showed low mood, decreased interest and energy, and gradually developed into manifestations of less eating, less talking, less movement, repeated psychiatric visits, and showed no improvement after treatment with antidepressants. The purpose of the current study is to increase the knowledge of these diseases and reduce misdiagnosis.
8.A case of postoperative obstructive hydrocephalus after pinealoma surgery onset with depressive symptoms
Meiti WANG ; Chongze WANG ; Ni ZHOU ; Yupeng CHEN ; Yinghua TAO ; Fang WANG ; Qinyu LYU ; Wu HONG
Chinese Journal of Psychiatry 2023;56(6):453-456
Pinealoma often comorbid with obstructive hydrocephalus due to their specific location and type, therefore, patients with pinealoma need to be treated by a combined ventriculoperitoneal shunt (VPS) along with surgical resection. Shunt failure is the most common complication after surgery. In this paper, we report a case of obstructive hydrocephalus caused by shunt obstruction after pineal gland tumor resection combined with ventriculoperitoneal shunt. The patient first showed low mood, decreased interest and energy, and gradually developed into manifestations of less eating, less talking, less movement, repeated psychiatric visits, and showed no improvement after treatment with antidepressants. The purpose of the current study is to increase the knowledge of these diseases and reduce misdiagnosis.
9.Study on the consistency and correlation of different methods for measuring energy consumption in patients with severe acute pancreatitis
Jing LIU ; Yao WU ; Xin HUANG ; Huajing KE ; Yupeng LEI ; Wenhua HE ; Yin ZHU ; Nonghua LYU ; Liang XIA
Chinese Journal of Digestion 2022;42(6):378-382
Objective:To analyze the consistency and correlation of indirect calorimetry (IC) and Harris-Benedict estimation method (H-B) in measuring energy consumption in patients with severe acute pancreatitis (SAP), as well as the characteristics of energy metabolism at resting state of SAP patients with different etiologies, and so as to guide the formulation of clinical energy nutrition support program.Methods:From February 1 to December 31, 2019, 61 SAP patients admitted into the intensive care unit of the First Affiliated Hospital of Nanchang University were enrolled. Collected relevant data of patients, such as the etiology of SAP, whether mechanical ventilation was needed, as well as the metabolic status. All SAP patients accepted IC test on the first day after admission to intensive care unit. According to the energy consumption measured by IC (hereinafter referred to as IC value) and the energy consumption measured by H-B (hereinafter referred to as H-B value), SAP patients were determined to be in high, normal or low metabolic state. Bland-Altman method and Pearson linear regression were used to analyze the consistency and correlation of the two methods in measuring energy consumption, and the linear equation was fitted. Paired t test was used for statistical analysis. Results:Among the 61 SAP patients, 13 cases were biliary SAP, 30 cases were hyperlipidemic SAP, 6 cases were alcoholic SAP, and 12 cases were SAP of other causes or unknown causes. Nineteen cases needed mechanical ventilation and 42 cases did not need mechanical ventilation. There were 70.5% (43/61) of SAP patients in high metabolic state, 13.1%(8/61) of SAP patients in normal metabolic state, and 16.4% (10/61) of SAP patients in low metabolic state. The IC value in 61 SAP patients was higher than H-B value((8 604.7±367.8) kJ/d vs. (6 491.2±133.7) kJ/d), and the difference was statistically significant ( t=5.95, P<0.001). The IC value in patients with hyperlipidemic, alcoholic and biliary SAP was (8 815.2±537.9), (7 631.2±890.5), and (8 108.0±933.1) kJ/d, respectively, and the H-B value was (6 869.6±204.5), (5 916.8±153.7), and (5 974.2±200.9) kJ/d, respectively. The IC value in patients with biliary or hyperlipidemic SAP were higher than H-B value, and the differences were statistically significant( t=2.29 and 3.38, both P<0.05). The IC value and H-B value in SAP patients with mechanical ventilation and without mechanical ventilation were (10 485.0±741.1) and (6 462.6±222.8) kJ/d, (6 595.1±364.7) and (6 503.2±166.7) kJ/d, respectively. The IC value in SAP patients with mechanical ventilation was higher than H-B value and the IC value in SAP patients without mechanical ventilation, and the differences were statistically significant ( t=4.71 and 5.20, both P<0.001). The consistency analyzed by Bland-Altman method showed that the two methods had obvious biases, and the average bias value was 92.2 kJ/d. The result of Pearson linear regression analysis indicated that there was a linear relationship between two methods ( r=0.44, P<0.001). The linear equation fitted by the univariate regression was Y=1.240 6 X+ 154.42, in which X was the H-B value and Y was the IC value. Conclusions:SAP patients are mostly in a hypermetabolic state in the early stages of the disease. The IC value is higher than H-B value, and it is recommended to use IC to measure energy consumption in SAP patients.
10.Drug resistance and virulence characteristics of of Staphylococcus aureus isolated from skin sites of suppurative infections in Inner Mongolia
Jianlin SUN ; Yang ZHANG ; Yupeng ZHANG ; Xinxiang LYU ; Junrui WANG
Chinese Journal of Dermatology 2022;55(3):200-207
Objective:To analyze drug resistance, virulence and molecular epidemiological characteristics of Staphylococcus aureus ( S. aureus) isolated from skin sites of suppurative infections, and to provide an experimental basis for clinical anti-infective therapies. Methods:Swab samples from suppurative skin lesions and nasal secretions were collected from inpatients in Department of Dermatology, the Affiliated Hospital of Inner Mongolia Medical University from May 2020 to December 2020, and subjected to bacterial isolation and culture. Suspected S. aureus colonies were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Drug sensitivity test was conducted by using the broth microdilution method. Virulence genes of S. aureus were amplified by PCR, and real-time fluorescence-based quantitative PCR was performed to determine the relative expression of 4 virulence genes including tsst-1, pvl, hla and clfA in S. aureus strains from different sources. S. aureus strains were genotyped by multilocus sequence typing. Drug resistance rates and detection rates of virulence genes were compared by using chi-square test or Fisher′s exact test, and measurement data among groups were compared by using t test or Mann-Whitney U test. Results:A total of 85 strains of S. aureus were isolated from 210 inpatients, including 54 isolates from skin sites of suppurative infections (case group) and 31 isolates from the nasal cavity (control group) . Drug sensitivity test showed that 14 strains of methicillin-resistant S. aureus (MRSA) were identified among 85 strains of S. aureus. The resistance rate to penicillin was the highest (90.59%, 77/85) in the 85 S. aureus strains; the resistance rates to clindamycin and erythromycin were 60.00% (51/85) and 61.18% (52/85) respectively; no strains showed resistance to rifampicin, vancomycin or linezolid. PCR showed that the detection rate of the pvl gene was 33.33% (18/54) in the case group, which was significantly higher than that in the control group (12.90%, 4/31; χ2= 4.28, P= 0.038) . Real-time fluorescence-based quantitative PCR showed that the relative expression level of the clfA gene was significantly higher in the control group (3.87[2.30, 5.94]) than in the case group (1.63[0.95, 2.62], P= 0.007) . A total of 17 ST types were identified among the 85 strains of S. aureus, and the dominant types were ST398-methicillin-susceptible S. aureus (20/71) and ST22-MRSA (9/14) . The detection rate of the virulence gene pvl was significantly higher in the ST22-MRSA strain (14/14) than in the non-ST22 MRSA strains (0, P < 0.001) . Conclusions:S. aureus strains isolated from the skin sites of suppurative infections were highly resistant to penicillin, clindamycin and erythromycin, so these antibiotics should not be used as the first-choice empiric treatment. The occurrence of cutaneous S. aureus infections may be associated with the virulence gene pvl, and the nasal colonization of S. aureus may be associated with the clfA gene.

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