1.Establishment of a method for acquisition, perfusion, preservation and transportation of the genetically modified donor pig kidneys
Feiyan ZHU ; Yaobo ZHAO ; Hongfang ZHAO ; Taiyun WEI ; Wenjie CHENG ; Kai LIU ; Yuexiao BAO ; Yaling LOU ; Hongjiang WEI ; Kaixiang XU
Organ Transplantation 2025;16(2):272-279
Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.
2.The management of infantile laryngeal cysts.
Fan LOU ; Cheng MING ; Yan GAO ; Jinyan ZU ; Jing MA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(2):120-123
Objective:To explore the therapeutic strategy for laryngeal cysts in infants. Methods:A retrospective analysis of the clinical data of 19 children with laryngeal cysts treated in Kunming Children's Hospital from January 2020 to January 2023. All patients were diagnosed through electronic laryngoscopy examination. Twelve neonates were admitted to the neonatal intensive care unit. Five of them received mechanical ventilation with tracheal intubation before surgery due to severe respiratory distress, and seven received oxygen therapy with a head mask. The remaining seven children were admitted to Department of Otolaryngology Head and Neck Surgery, of which three cases were treated with oxygen therapy through a mask during sleep due to frequent shortness of breath during sleep. All patients underwent low-temperature plasma radiofrequency ablation under general anesthesia to remove the cysts. Results:Three newborns were unable to have their tracheal tubes removed due to complications with pneumonia after surgery, while the rest of the children were able to have their tubes successfully removed after complete anesthesia, and no gastric tubes were placed. All postoperative respiratory difficulties in the children disappeared, and there were no complications such as bleeding, hoarseness, or laryngeal stenosis. Five pediatric patients had incomplete relief of laryngeal ringing symptoms one month after surgery, and electronic laryngoscopy diagnosed laryngeal softening. Regular follow-up is recommended. One child relapsed 4 months after surgery and underwent a follow-up surgery six months later without recurrence. Conclusion:Endoscopic low-temperature plasma radiofrequency ablation is an effective surgical method for treating laryngeal cysts, with a low postoperative recurrence rate. Laryngeal cysts may be accompanied by laryngeal softening, which may be a possible reason for the postoperative symptoms not improving.
Humans
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Retrospective Studies
;
Cysts/surgery*
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Laryngeal Diseases/surgery*
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Infant
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Laryngoscopy
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Infant, Newborn
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Male
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Female
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Radiofrequency Ablation
3.Clinical trial of salmeterol and fluticasone propionate combined with tiotropium in treatment of elderly patients with stable COPD
Xia ZHOU ; LI-Li LOU ; Liu YANG ; Hui-Xin CHENG
The Chinese Journal of Clinical Pharmacology 2024;40(17):2459-2463
Objective To analyze the efficacy of salmeterol rotticasone combined with tiotropium bromide in the treatment of elderly patients with stable chronic obstructive pulmonary disease(COPD)and its influence on serum procalcitonin(PCT),serum amyloid A(SAA)and albumin-to-fibrinogen ratio(AFR)levels.Methods The elderly patients with stable COPD were divided into control group and treatment group.The control group was treated with salmeterol rotticasone inhalation powder(1 inhalation each time,twice a day),while the treatment group was given tiotropium bromide inhaled powder aerosol treatment(1 inhalation each time once a day)on the basis of the control group.Patients in two groups were treated for 3 months.The clinical efficacy and differences in pulmonary function[forced expiratory volume in one second(FEV1),FEV1/forced vital capacity(FEV1/FVC),peak expiratory flow(PEF)],modified British Medical Research Council respiratory questionnaire(mMRC)score,COPD Assessment Test(CAT)score and serum PCT,SAA and AFR levels were compared;and the safety was evaluated.Results Fifty-two patients and fifty-eight patients were included in treatment group and control group,respectively.After treatment,the total effective rates in treatment group and control group were 90.38%(47 cases/52 cases)and 75.86%(44 cases/58 cases)(P<0.05).The FEV1 values in treatment group and control group were(1.49±0.33)and(1.36±0.29)L;the FEV1/FVC values were(61.28±10.56)%and(57.41±10.03)%;PEF values were(5.32±0.88)and(4.93±0.75)L·s-1;mMRC scores were(0.99±0.24)and(1.21±0.27)points;the CAT scores were(11.58±3.02)and(14.26±3.71)points;PCT levels were(0.76±0.11)and(0.85±0.18)μg·L-1;SAA levels were(34.59±4.15)and(37.66±5.34)mg·L-1;AFR levels were 5.97±1.12 and 5.53±1.06,respectively.Compared with the control group,the above indexes in the treatment group were statistically significant(all P<0.05).The main adverse drug reactions in treatment group were dry mouth,constipation and dizziness and nausea,and the main adverse drug reactions in control group were dry mouth,constipation,and dizziness and nausea.The total incidence rates of adverse drug reactions in treatment group and control group were 21.15%(11 cases/52 cases)and 13.79%(8 cases/58 cases)respectively,with no statistical significance(P>0.05).Conclusion Salmeterol ticasone inhalation powder and tiotropium bromide inhaled powder aerosol can effectively enhance the clinical efficacy of elderly patients with stable COPD,improve the pulmonary function,relieve the body's inflammation,and promote the improvement of disease condition,with good safety.
4.Determination of Anlotinib in Human Plasma by UPLC-MS/MS and Its Clinical Application
Jiang LOU ; Hong JIANG ; Junjie LAO ; Ling CHEN ; Cheng YI ; Xinmei WU ; Jing WANG ; Gang WANG
Chinese Journal of Modern Applied Pharmacy 2024;41(11):1515-1523
OBJECTIVE
To establish a ultra-high-performance liquid chromatography-mass spectrum/mass spectrum(UPLC-MS/MS) method for the determination of anlotinib in human plasma and assessment of clinical application.
METHODS
Zanubrutinib was used as internal standard and the extraction process was performed through protein precipitation method using acetonitrile, followed by separation on an Ultimate XB-C18(100 mm×2.1 mm, 3.0 μm) column using acetonitrile and 10 mmol·L−1 ammonium acetate-0.1% formic acid step-elution gradient. The flow rate was 0.6 mL·min−1 and injection volume was 5 μL. The mass analysis was performed by positive ion electrospray ionization in multiple-reaction monitoring mode, and the mass spectrometer was set at m/z 408.1→339.1 for anlotinib and m/z 472.2→290.1 for internal standard, respectively. The specificity, standard curve and lower limit of quantification, precision and recovery, matrix effect and stability of the method and clinical application were investigated.
RESULTS
The method was validated over the concentration range of 1.0−100.0 ng·mL−1, with R2=0.998 4. The precision RSD was<9%, the recovery and matrix effect were 104.81%−107.32% and 102.54%−105.26%, respectively, and this method had good stability and was not affected by matrix effect. The method had been used for determined 52 advanced non-small cell lung cancer patients treated with anlotinib. The trough plasma concentration (Ctrough) was measured on day 43 after initiation of anlotinib treatment. Anlotinib Ctrough were higher than lower limit of quantitation (1.0 ng·mL−1) from 52 patients. The plasma concentration of anlotinib Ctrough was (11.38±4.29)ng·mL−1 with 37.66% coefficients of variation, which were shown large inter-patient variability.
CONCLUSION
This method is high sensitivity, specificity and accurate, and suitable for determination of anlotinib in human plasma.
5.Treatment of chronic sinus tract leakage at rectal anastomosis with anal fistula endoscopy
Liqiang JI ; Jialing ZHOU ; Cheng XIN ; Shuyuan LI ; Ye WANG ; Zheng LOU ; Wei ZHANG
Chinese Journal of Gastrointestinal Surgery 2024;27(10):1080-1082
Objective:To introduce the method of using anal fistula endoscope to treat chronic sinus tract leakage at rectal anastomosis site.Methods:We used anal fistula endoscopy to treat a patient with chronic sinus tract leakage after radical resection of rectal cancer, mainly including the following 5 steps: (1) establishing a water injection circulation system through the anus; (2) scraping off purulent coating and mucosa on the surface of the sinus tract with the brush; (3) hemostasis and removal of necrotic tissue with electrocoagulation rods; (4) filling the sinus tract with bioprotein gel; (5) compressing the sinus tract with transanal drainage tube.Results:The patient is a 70 year old male with rectal cancer. After undergoing 3D laparoscopic assisted radical resection of rectal cancer via abdominal anterior resection (Dixon's procedure) and diverting ileostomy surgery for more than 3 months, leakage of the rectal anastomosis was found through colonoscopy and anal iodine water contrast imaging .The patient started eating and flowing juice 6 hours after surgery, got out of bed 24 hours after surgery, and was discharged 48 hours after the removal of the anal canal. Three months after surgery, colonoscopy and transanal iodine hydrography showed that the sinus repair was intact. The diverting ileostomy was reduced 4 months after surgery.Conclusion:Anal fistula endoscope is safe and feasible for the treatment of chronic sinus tract anastomotic leakage in selected patients.
6.Influencing factors of futile recanalization after endovascular intervention in patients with acute basilar artery occlusion
Yi YAN ; Kemeng ZHANG ; Wansi ZHONG ; Shenqiang YAN ; Bing ZHANG ; Jianhua CHENG ; Min LOU
Journal of Zhejiang University. Medical sciences 2024;53(2):141-150
Objective:To explore the influence factors for futile recanalization following endovascular treatment(EVT)in patients with acute basilar artery occlusion(BAO).Methods:Clinical data of patients with acute BAO,who underwent endovascular treatment within 24 h of onset from January 2017 to November 2022,were retrospectively analyzed.The futile recanalization was defined as modified thrombolysis in cerebral infarction(mTICI)grade≥2b or 3 after successful reperfusion,but the modified Rankin Scale score>2 at 3 months after EVT.Binary logistic regression model was used to analyze the influencing factors of futile recanalization.Results:A total of 471 patients with a median age of 68(57,74)years were included and 68.9%were males,among whom 298(63.27%)experienced futile recanalization.Multivariate analysis revealed that concomitant atrial fibrillation(OR=0.456,95%CI:0.282-0.737,P<0.01),bridging thrombolysis(OR= 0.640,95%CI:0.416-0.985,P<0.05),achieving mTICI grade 3(OR=0.554,95%CI:0.334-0.918,P<0.05),arterial occlusive lesion(AOL)grade 3(OR=0.521,95%CI:0.326-0.834,P<0.01),and early postoperative statin therapy(OR=0.509,95%CI:0.273-0.948,P<0.05)were protective factors for futile recanalization after EVT in acute BAO patients.High baseline National Institutes of Health Stroke Scale(NIHSS)score(OR=1.068,95%CI:1.049-1.087,P<0.01),coexisting hypertension(OR=1.571,95%CI:1.017-2.427,P<0.05),multiple retrieval attempts(OR=1.237,95%CI:1.029-1.488,P<0.05)and postoperative hemorrhagic transformation(OR=8.497,95%CI:2.879-25.076,P<0.01)were risk factors.For trial of ORG 10172 in acute stroke treatment(TOAST)classification,cardiogenic embolism(OR=0.321,95%CI:0.193-0.534,P<0.01)and other types(OR=0.499,95%CI:0.260-0.961,P<0.05)were related to lower incidence of futile recanalization.Conclusions:The incidence of futile recanalization after EVT in patients with acute BAO is high.Bridging venous thrombolysis before operation and an early postoperative statin therapy may reduce the incidence of futile recanalization.
7.Effect of anesthesia mode during endovascular treatment on neurological functional outcomes in patients with acute posterior circulation stroke
Xiaofeng ZHU ; Zheyu ZHANG ; Wansi ZHONG ; Yaode HE ; Zhongyu LUO ; Ningyuan ZHANG ; Chaochan CHENG ; Jianhong YANG ; Min LOU
Journal of Zhejiang University. Medical sciences 2024;53(2):151-159
Objective:To compare the effect of anesthesia mode on the neurological functional outcomes in patients undergoing endovascular treatment for acute posterior circulation ischemic stroke.Methods:Clinical data of 656 patients undergoing intravascular therapy for acute posterior circulation ischemic stroke registered in online Acute Stroke Patients for Stroke Management Quality Evaluation Database from January 2017 to December 2022 were retrospectively analyzed.The data included 163 cases with conscious sedation and 493 cases with general anesthesia during the procedure.After propensity score matching,428 patients were included in the analysis,including 155 cases in the conscious sedation group and 273 cases in the general anesthesia group.The differences of operation mode,etiology type,vascular recanalization,hemorrhagic transformation at 24 h,modified Rankin Scale(mRS)score at 3 months and mortality within 3 months were compared between the two groups.Binary logistic regression was used to explore the effect of different anesthesia mode on neurological functional outcomes.Results:There was a significant difference in operation mode between the two groups(P<0.01),while there were no significant differences in etiology type,vascular recanalization,hemorrhagic transformation at 24 h,mRS score at 3 months or mortality within 3 months(all P>0.05).Binary logistic regression analysis revealed that anesthesia modes were not significantly associated with functional outcomes of patients(OR=1.151,95%CI:0.751-1.765,P>0.05).Conclusion:Anesthesia mode(conscious sedation or general anesthesia)will not affect the neurological functional outcomes in patients with acute posterior circulation ischemic stroke undergoing endovascular treatment.
8.Analysis of perioperative risk factors in young children with obstructive sleep apnea
Fan LOU ; Cheng MING ; Jinyan ZU ; Liping ZHAO ; Jing MA
Chinese Archives of Otolaryngology-Head and Neck Surgery 2024;31(5):326-329
OBJECTIVE To explore the perioperative risk factors in young children with obstructive sleep apnea.METHODS The 86 young OSA children admitted to Kunming Children's Hospital from January 2020 to December 2022 were divided into general ward group and ICU ward group according to their postoperative treatment.The clinical data of the two groups were compared and analyzed.RESULTS The course of disease and operation time of children in ICU ward group were significantly longer than those in general ward group,OAHI and ODI were significantly greater than those in general ward group,the intraoperative blood loss was significantly more than that in general ward group,MSaO2 and LSaO2 were significantly lower than those in general ward group,and the tonsil size and operation method composition ratio were significantly different from those in general ward group(P<0.05).There were no significant differences in sex composition ratio,age,weight,height,BMI and adenoid size grading ratio between the two groups(P>0.05).The OAHI values of the two groups were significantly negatively correlated with MSaO2 and LSaO2(r=-0.676,-0.724),and significantly positively correlated with tonsil size grade,ODI,operation time and intraoperative blood loss(r=0.242,0.967,0.321,0.446,P<0.05).There was no significant correlation with the course of disease(r=0.172,P>0.05).Multiple linear regression analysis showed that LSaO2 and ODI were independent risk factors for the severity of the child's condition.CONCLUSION The severity of the condition in young OSA children determines the perioperative risk and is influenced by the type of surgery.LSaO2 and ODI are independent risk factors which should be taken seriously by clinicians.
9.Application of Apoptosis Small-Molecule Probe in Acute Middle Cerebral Artery Embolization and Recanalization Model
Cheng QIAN ; Tao WANG ; Yinghao LI ; Wensheng LOU ; Jianping GU ; Guoping CHEN
Chinese Journal of Medical Imaging 2024;32(10):977-981,987
Purpose To investigate the feasibility of apoptosis small-molecule probe CYS-F for in vivo molecular imaging in acute middle cerebral artery embolization and recalculation models,and to explore its ability to reflect lesions.Materials and Methods Hela cell apoptosis was induced by Adriamycin,and the targeting ability of the probe was verified in vitro.Clinical middle cerebral artery embolization was simulated,acute ischemic stroke model(n=15)and recanalization model(n=15)were constructed,and the vessels were evaluated by magnetic resonance angiography imaging.The distribution of CYS-F was investigated by injecting the probe into the caudal vein.24 hours after modeling,the lesion volume rate was assessed on T2WI,and apoptosis was assessed in vivo by near infrared imaging.Niss and c-fos staining were used to compare the differences between the two groups.Results CYS-F showed a good ability to target apoptosis in vitro.After modeling,Doppler flowmeter and magnetic resonance angiography showed that the middle cerebral artery was successfully embolized in the embolization group,and the artery was recanalized in the recanalization group.Near infrared imaging showed the loss of fluorescence signal in the middle cerebral artery region in the embolized group.24 hours after modeling,T2WI showed that the infarct volume rate in the recanalization group was significantly lower than that in the embolization group(0.055±0.015 vs.0.512±0.220;t=19.761,P<0.001).The fluorescence intensity of the embolization group was significantly stronger than that of the recusing group,the target to background ratio was 1.215±0.162 and 0.731±0.085,respectively(t=10.252,P<0.001).In the embolization group,a large number of activated neurons expressed c-fos protein,and a large number of cells underwent nuclear condensation and lysis by Niss staining.Conclusion The acute embolization model and recalculation model of middle cerebral artery in mice are close to the clinic.CYS-F can be used to image apoptosis in vivo,reflect the lesion degree,and reflect the local tissue blood perfusion at the initial stage of injection.
10.Clinical efficacy of AcoStream peripheral thrombus aspiration system combined with catheter-directed thrombolysis in treatment of lower extremity deep vein thrombosis
Tao WANG ; Wensheng LOU ; Haobo SU ; Cheng QIAN ; Yinghao LI ; Guoping CHEN ; Xu HE ; Jianping GU
Chinese Journal of Radiology 2024;58(5):523-528
Objective:To investigate the efficacy and safety of AcoStream peripheral thrombus aspiration system combined with catheter-directed thrombolysis in the treatment of lower extremity deep vein thrombosis.Methods:The clinical data of 16 lower extremity deep vein thrombosis cases treated with AcoStream peripheral thrombus aspiration system combined with catheter-directed thrombolysis, admitted to the authors′ hospital from May 2022 to November 2022, were retrospectively analyzed. The differences in circumferential diameter between the affected limb and the healthy side, venous patency score, thrombus clearance grade and intraoperative blood loss were observed and compared. The Villalta score was used during the follow up. Paired sample t-test and Wilcoxon rank sum test were used to compare the changes in the observed indicators before and after treatment to evaluate the efficacy. Results:Treatment were successfully performed in all patients. Before treatment, the circumference differences between the affected and unaffected thighs and calves were (3.69±0.97) and (3.34±0.75)cm, respectively, the venous patency score of the affected side was 8(7.25,9) points. After treatment, the circumference differences between the affected and unaffected thighs and calves were (0.81±0.68) and (0.84±0.70)cm, respectively. The venous patency score of the affected side was 1(0,1)points, with statistically significant differences ( P<0.001). Grade Ⅲ thrombus clearance was achieved in 7 patients, grade Ⅱ thrombus clearance was achieved in 9 patients. The average blood loss during thrombus aspiration was (133.1±12.0) ml. Following up for 6 months, the Villalta score was 0(0,1.75) points. Conclusion:Acostream peripheral thrombus aspiration system combined with catheter-directed thrombolysis is safe and effective for the treatment of lower extremity deep venous thrombosis, with satisfactory short-term efficacy and high clinical application value.


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