1.Effects of Sanhuang Gel on p38MAPK Signaling Pathway and Its Key Molecules in Rat Auricle Acne Complex Model
Dan LI ; Xueyang DU ; Sinong WANG
Chinese Journal of Modern Applied Pharmacy 2024;41(8):1035-1046
OBJECTIVE
To observe the anti-inflammatory mechanism of Sanhuang gel on rat model of auricle acne and its possible mechanism with p38MAPK signaling pathway as the entry point.
METHODS
Sixty-two male Wistar rats were divided into blank group and modelling group. The modeling group replicated the experimental animal model of acne(application of 100% oleic acid+subcutaneous injection of Propionibacterium acnes), and 2 rats were randomly selected for HE staining after 21 days. The remaining rats were randomly divided into model group, positive control group(Vitamin A cream 1.48 g·d−1 ), Sanhuang gel high, medium and low-dose group(1.1, 0.568, 0.3 g·d−1 ). They were administered for 8 weeks, and observed on the 7, 14 and 28 day of treatment. The right auricle tissues were collected, and the histopathological changes were observed by HE stain. The mRNA expressions of p38MAPK, MKK3 and MKK6 were detected by RT-PCR. The protein expressions of MCP-1, HMGB1 and α-SMA in auricle tissues were detected by IHC and Western blotting.
RESULTS
Compared with blank group, the epidermis of auricular tissue in model group was severely keratinized, inflammatory cell infiltration was obvious, mRNA levels of p38MAPK, MKK3, MKK6 and protein expression levels of MCP-1, HMGB1 and α-SMA were increased(P<0.05). After drug therapy intervention, auricle apparent score at different time points in each treatment group was improved to varying degrees. The mRNA levels of p38MAPK, MKK3, MKK6 and protein levels of MCP-1, HMGB1 and α-SMA in Sanhuang gel high-dose and medium-dose groups and positive control group were decreased(P<0.05), while in Sanhuang gel low-dose group, the expressions of MKK3 mRNA and MCP-1, HMGB1 protein were decreased(P<0.05), the expression of p38MAPK, MKK6 mRNA and α-SMA protein decreased not significantly.
CONCLUSION
The mechanism of the effect of Sanhuang gel on the inflammatory response of rat compound acne animal model may be related to its intervention of p38MAPK protein feedback regulation signaling pathway and inhibition of MCP-1, HMGB1 and α-SMA expression.
2.MRI study of the relationship between the cerebral small vessel disease total burden and imaging markers and degree of middle cerebral artery stenosis
Xinbo XING ; Xueyang WANG ; Jinhao LYU ; Qi DUAN ; Caohui DUAN ; Xiangbing BIAN ; Kun CHENG ; Mingliang YANG ; Tingyang ZHANG ; Chenglin TIAN ; Xin LOU
Chinese Journal of Radiology 2024;58(1):34-40
Objective:To investigate the relationship between the cerebral small vascular disease (CSVD) total burden and the imaging markers and the degree of unilateral middle cerebral artery (MCA) stenosis.Methods:The study was a cross-sectional study. Clinical and imaging data of patients with chronic unilateral MCA stenosis who underwent multimodal MRI from October 2015 to January 2019 in the First Medical Center of PLA General Hospital were retrospectively analyzed. A total of 261 patients were included, 187 males and 74 females. According to the degree of MCA stenosis, the patients were divided into 102 cases in severe stenosis-occlusion group (stenosis degree ≥70%) and 159 cases in mild-moderate stenosis group (stenosis degree <70%). CSVD imaging marker scores (including white matter hyperintensity, perivascular space, cerebral microbleed, and lacune of presumed vascular origin) were assessed according to the ?standards for reporting vascular changes on neuroimaging 1 in the 2 groups, and the CSVD total burden score was calculated. Mann-Whitney U test was used to compare the indicators between the two groups, and the CSVD total burden score and imaging marker scores were ultimately included in a multifactorial binary logistic regression to assess the association of CSVD imaging markers with severe stenosis-occlusion of the MCA after adjusting for vascular risk factors (age, gender, drinking, smoking, hypertension, hyperlipidemia, atrial fibrillation and coronary heart disease). Results:There were significant differences in the CSVD total burden, centrum semiovale perivascular space and lacune of presumed vascular origin score between the mild-to-moderate stenosis group and the severe stenosis-occlusion group (all P<0.05), and none of the differences in the remaining imaging marker scores were statistically significant (all P>0.05). Multivariate binary logistics regression analysis showed CSVD total burden score ( OR=1.300, 95% CI 1.047-1.613, P=0.017), centrum semiovale perivascular space score ( OR=2.099, 95% CI 1.540-2.860, P<0.001) and lacune of presumed vascular origin score ( OR=2.609, 95% CI 1.294-5.261, P=0.007) were independent associated with severe stenosis-occlusion of MCA. Conclusion:The higher CSVD total burden score, centrum semiovale perivascular space score and lacune of presumed vascular origin score are associated with severe stenosis-occlusion of MCA.
3.Long-term donor safety analysis after related renal transplantation in a single center
Jiyuan WANG ; Yu CHEN ; Jinghui YANG ; Xueyang ZHENG ; Yue DING ; Shu HAN
Academic Journal of Naval Medical University 2024;45(11):1390-1394
Objective To evaluate the long-term safety of related kidney donors after unilateral nephrectomy.Methods A total of 91 related donors who received nephrectomy in our hospital from 2006 to 2011 were followed up for at least 10 years by outpatient,telephone,or WeChat.During the follow-up period,the serum creatinine,serum uric acid,blood urea nitrogen,estimated glomerular filtration rate(eGFR),hematuria,urinary protein,blood pressure,blood glucose and blood lipids of the donors were detected,and the changes before and after nephrectomy were analyzed.Results At 1 month after operation,the levels of serum creatinine,blood urea nitrogen and serum uric acid of the donor were significantly higher than those before operation(all P<0.05),but still within the normal range.The patients were followed up for 1,3,5 and 10 years after operation.Compared with 1 month after operation,the serum creatinine,blood urea nitrogen and serum uric acid were relatively stable(all P>0.05).The eGFR of donors of different ages remained relatively stable for a long time after operation.There were 3 cases of endoscopic hematuria and 4 cases of proteinuria after surgery,and these symptoms were relieved after rest and symptomatic treatment.Ten(11.0%)donors developed hypertension 5(5.5%)developed hyperlipidemia,and 5(5.5%)developed diabetes mellitus.No patient died.Conclusion Nephrectomy is safe and feasible for healthy related donors.To ensure the safety of the donors,comprehensive evaluation before nephrectomy and regular follow-up after nephrectomy are essential.
4.Clinical characteristics and risk factors of complications after kidney transplantation in children at a single-center
Fanyuan ZHU ; Xueyang ZHENG ; Jinghui YANG ; Jiyuan WANG ; Yue DING ; Yu CHEN ; Shu HAN
Chinese Journal of Organ Transplantation 2024;45(6):391-398
Objective:To explore the clinical characteristics and risk factors of pediatric kidney transplantation (KT).Methods:From January 1, 2010 to September 30, 2022, retrospective analysis was performed for the relevant clinical data of 81 pediatric recipients of primary KT at Organ Transplant Center of Shanghai Changzheng Hospital. The occurrences of acute rejection (AR) ,delayed graft function (DGF), infection, myelosuppression, tumor and other complications were observed within 1 year post-KT. They were grouped according to whether or not AR/DGF occurred. Univariate analysis speculated the effect of AR and DGF on renal function at 1 year after transplantation. Binary Logistic regression was employed for examining the risk factors related to AR/DGF.Results:During follow-ups, transplanted kidney was removed due to an embolization of renal vessels and dialysis resumed (n= 5). One child had failed graft due to the recurrence of original disease and dialysis resumed. The remaining 75 children had an excellent recovery of graft function. At the end of follow-ups, survival for transplant recipients and transplanted kidneys was 100% (81/81 ) and 92.6% (75/81) respectively. 23 patients (28.4%) developed DGF, including 20 child recipients of C-I donors. Among DGF recipients, 21 (91.3%) were immune induced with anti-CD25 humanized monoclonal antibody and 2 (8.7 %) with porcine antihuman lymphocyte immunoglobulin (pALG). Within the first year post-KT, 13 patients (16.1%) developed AR, including 11 child recipients of C-I donors. Induction was made with anti-CD25 humanized monoclonal antibody (n=8), pALG (n=4) and anti-human T lymphocyte rabbit immunoglobulin (n=1). And 12 cases were reversed with MP (methylprednisolone) shock therapy while another ineffective case was rescued by an intravenous infusion of rATG (rabbit anti-human thymocyte immunoglobulin). During postoperative follow-ups, 14 (17.3 %) KT recipients had an onset of pulmonary infection (n=7), upper respiratory tract infection (n=3), urinary tract infection (n=5), gastrointestinal infection (n=2) and abdominal cavity infection (n=1). The causative pathogens were bacteria (n=14) and viruses (n=4). Among 7 cases (8.6%) of myelosuppression, there were leukopenia (n=6) and thrombocytopenia (n=1 ). During 1-year follow-ups, no malignancy occurred. At the last follow-up, blood creatinine was (72.79±21.07) μmol/L in non-AR/DGF recipients. For AR/DGF recipients, blood creatinine levels were (68.83±10.78) and (74.20±18.70) μmol/L. There was no significant inter-group difference ( F=0.14, P=0.87). In groups with and without DGF, the incidence of bone marrow suppression in the children with DGF was significantly higher (21. 74 %) than that in the untreated group (3.45%), with a statistically significant difference ( P=0.02). However, there was no statistically significant difference in the age, sex, donor source, infection, and types of immune-induced drugs in AR, DGF occurrence and no occurrence group. logistic Regression analysis showed that immunoinduction therapy with lymphocyte inhibitor ( OR=0.074, 95 %CI: 0.009-0.0643, P=0.018) and bone marrow suppression ( OR=0.045, 95%CI: 0.004-0.515, P=0.013) were risk factors for DGF. Conclusion:KT in children may obtain decent outcomes. Immunoinduction therapy with lymphocyte inhibitors and occurrence of myelosuppression are risk factors for postoperative DGF. The occurrence of AR/DGF in early postoperative period does not affect the level of kidney function in children at 1 year post-KT. It is recommended to closely follow up and accumulate experiences for optimizing long-term outcomes.
5.Application of fast susceptibility weighted imaging based on deep learning in assessment of acute ischemic stroke
Qi DUAN ; Caohui DUAN ; Shiqing ZHOU ; Jinhao LYU ; Xiangbing BIAN ; Dekang ZHANG ; Kun CHENG ; Mingliang YANG ; Xueyang WANG ; Tingyang ZHANG ; Xinbo XING ; Chenglin TIAN ; Xin LOU
Chinese Journal of Radiology 2023;57(1):34-40
Objective:To explore the value of fast susceptibility weighted imaging (SWI) generated by a deep learning model in assessment of acute ischemic stroke (AIS).Methods:From January 2019 to January 2021, 118 AIS patients [75 males and 43 females, aged 23-100 (66±14) years] who underwent MR examination and SWI sequence scanning within 24 h of symptom onset in the First Medical Center of PLA General Hospital were retrospectively analyzed. MATLAB ′s randperm function was used to divide 118 patients into a training set of 96 cases and a test set of 22 cases at a ratio of 8∶2. Fourty-seven AIS patients [38 males and 9 females, aged 16-75 (58±12) years] from one center of a multicenter study were selected to build the external validation set. SWI image and filtered phase image were combined into complex value image as full sampling reference image. Undersampled SWI images were obtained by retrospective undersampling of reference fully sampled images, and the undersampling multiple was five times which could save 80% of the scanning time, then the complex-valued convolutional neural network (ComplexNet) was used to develop reconstruct fast SWI. Interclass correlation coefficient (ICC) or Kappa tests were used to compare the consistency of image quality and the diagnostic consistency for the presence of susceptibility vessel sign (SVS), cerebral microbleeds and asymmetry of cerebral deep medullary veins (DMVs) in AIS patient on fully sampled SWI and fast SWI based on ComplexNet.Results:In test set, score of image quality was 4.5±0.6 for fully sampled SWI image and 4.6±0.7 for fast SWI based on ComplexNet, and coefficient was excellent (ICC=0.86, P<0.05). Full sampling SWI had good agreement with fast SWI based on ComplexNet in detecting SVS (Kappa=0.79, P<0.05), microbleeds (Kappa=0.86, P<0.05), and DMVs asymmetry (Kappa=0.82, P<0.05) in AIS patients. In the external validation set, score of image quality was 4.1±1.0 for fully sampled SWI image and 4.0±0.9 for fast SWI based on ComplexNet, and coefficient was excellent (ICC=0.97, P<0.05). Full sampling SWI had good agreement with fast SWI based on ComplexNet in detecting SVS (Kappa=0.74, P<0.05), microbleeds (Kappa=0.83, P<0.05), and DMVs asymmetry (Kappa=0.74, P<0.05) in AIS patients. Conclusions:Deep learning techniques can significantly accelerate the speed of SWI, and the consistency of image quality and detected AIS signs between fast SWI based on ComplexNet and fully sampled SWI is good. The fast SWI based on ComplexNet can be applied to the radiographic assessment of clinical AIS patients
6.Role of soy lecithin combined with soy isoflavone on cerebral blood flow in rats of cognitive impairment and the primary screening of its optimum combination
Hongrui LI ; Xianyun WANG ; Xiaoying LI ; Xueyang ZHOU ; Xuan WANG ; Tiantian LI ; Rong XIAO ; Yuandi XI
Nutrition Research and Practice 2023;17(2):371-385
BACKGROUND/OBJECTIVES:
Soy isoflavone (SIF) and soy lecithin (SL) have beneficial effects on many chronic diseases, including neurodegenerative diseases. Regretfully, there is little evidence to show the combined effects of these soy extractives on the impairment of cognition and abnormal cerebral blood flow (CBF). This study examined the optimal combination dose of SIF + SL to provide evidence for improving CBF and protecting cerebrovascular endothelial cells.MATERIALS/METHODS: In vivo study, SIF50 + SL40, SIF50 + SL80 and SIF50 + SL160 groups were obtained. Morris water maze, laser speckle contrast imaging (LSCI), and hematoxylineosin staining were used to detect learning and memory impairment, CBF, and damage to the cerebrovascular tissue in rat. The 8-hydroxy-2′-deoxyguanosine (8-OHdG) and the oxidized glutathione (GSSG) were detected. The anti-oxidative damage index of superoxide dismutase (SOD) and glutathione (GSH) in the serum of an animal model was also tested. In vitro study, an immortalized mouse brain endothelial cell line (bEND.3 cells) was used to confirm the cerebrovascular endothelial cell protection of SIF + SL. In this study, 50 μM of Gen were used, while the 25, 50, or 100 μM of SL for different incubation times were selected first. The intracellular levels of 8-OHdG, SOD, GSH, and GSSG were also detected in the cells.
RESULTS:
In vivo study, SIF + SL could increase the target crossing times significantly and shorten the total swimming distance of rats. The CBF in the rats of the SIF50 + SL40 group and SIF50 + SL160 group was enhanced. Pathological changes, such as attenuation of the endothelium in cerebral vessels were much less in the SIF50 + SL40 group and SIF50 + SL160 group. The 8-OHdG was reduced in the SIF50 + SL40 group. The GSSG showed a significant decrease in all SIF + SL pretreatment groups, but the GSH showed an opposite result. SOD was upregulated by SIF + SL pretreatment. Different combinations of Genistein (Gen)+SL, the secondary proof of health benefits found in vivo study, showed they have effective antioxidation and less side reaction on protecting cerebrovascular endothelial cell. SIF50 + SL40 in rats experiment and Gen50 + SL25 in cell test were the optimum joint doses on alleviating cognitive impairment and regulating CBF through protecting cerebrovascular tissue by its antioxidant activity.
CONCLUSIONS
SIF+SL could significantly prevent cognitive defect induced by β-Amyloid through regulating CBF. This kind of effect might be attributed to its antioxidant activity on protecting cerebral vessels.
7.Research Advances of Chemical Constituents and Analytical Methods of Shengmai Formula
Zixuan QIAN ; Xueyang SUN ; Chenming ZHANG ; Longchan LIU ; Linnan LI ; Haoyue ZHANG ; Li YANG ; Zhengtao WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2699-2708
Shengmai formula,composed of Ginseng Radix et Rhizoma,Ophiopogon Radix and Schisandrae Chinensis Fructus,is a classic and famous formula.It is a representative formula for"supplementing qi,nourishing yin,and generating fluid"in Traditional Chinese Medicine theory.To date,a wide range of Shengmai formulae have been developed according to different medical applications,but the quality evaluation standards are at a relatively low level,and most of them only specify the individual components of a single herb,making it difficult to ensure clinical efficacy and safety.At the same time,the physical and chemical identification methods of Shengmai formula have been constantly updated,allowing for greater progress in research on its main chemical components such as saponins,lignans and flavonoids.However,there is little systematic summarization of the chemical components and analytical methods.Based on the existing references,we systematically summarized ginsenosides,ophiopogonins,schisandra lignans,homoisoflavonoids and some other compounds in this paper,as well as the quality standards of Shengmai formulae and their analytical methods in order to aid clinical research and formulation manufacture.
8.Feasibility of a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection for middle and low rectal cancer
Wenjuan LI ; Dechang DIAO ; Jiaxin LIN ; Jiahao WANG ; Weilin LIAO ; Xin TANG ; Jiaxin XIE ; Lin AO ; Xueyang ZHANG ; Xiaojiang YI ; Xiaochuang FENG ; Hongming LI ; Xinquan LU
Chinese Journal of Gastrointestinal Surgery 2023;26(10):968-976
Objective:To explore the feasibility and value of performing a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection (LLND) for middle and low rectal cancer.Methods:This was a retrospective review. The study cohort comprised patients who met the diagnostic criteria for rectal cancer according to the Chinese Guidelines for the Diagnosis and Treatment of Colorectal Cancer, had a short lymph node diameter of >5 mm on the lateral side within the 15 days before surgery, were evaluated as feasible candidates for laparoscopic total mesorectal excision+LLND surgery, had been diagnosed with low or intermediate level rectal cancer, and whose tumor was less than 8 cm away from the anal verge according to pathological examination of the operative specimen. Patients with a history of other malignant tumors of the abdomen or with incomplete follow-up data were excluded. Forty-two patients with middle and low rectal cancer who had undergone lateral lymph node dissection in diagnosis and treatment center of Gastrointestinal Cancer of Guangdong Hospital of Chinese Medicine from Jan.2018 to Dec.2022 were enrolled. There were 24 men (57.1%) and 18 women (42.9%) aged 58.4±11.8 years and the median BMI was 22.5 (19.3–24.1) kg/m 2. The main point of the three-sided encapsulation procedure is to expand the external side medial to the external iliac artery and vein, narrowing the range of exterior side dissection. The anterior-medial side is designed to expand the vesical fascia to define the range of anterior-medial side extension. The internal side is fully extended to the ureterohypogastric nerve fascia; the distal point of the caudal extension reaches the level of the Alcock canal and the bottom reaches the piriformis, enabling dissection of the obturator nerve and No.283 lymph nodes. No.263D lymph nodes are dissected by exposing the internal iliac artery and its branches, dissecting the group No.263P lymph nodes, and severing the inferior vesical artery. Finally, the lateral lymphatic tissue is completely resected. Relevant variables were recorded, including the number of lateral lymph nodes detected, the rate of lymph node metastasis, operation duration, intraoperative blood loss, postoperative complications, postoperative hospital stay, and 3-year overall survival rate. Results:Laparoscopic surgery was successfully completed in all patients with no conversions to open surgery and no intraoperative complications. Twenty-seven (64.3%) of the study patients underwent left-sided LLND, 10 (23.8%) right-sided LLND, and five (11.9%) bilateral LLND, with lymph nodes cleared on both sides. All patients' lymph nodes were examined pathologically. A median of 17.0 (11.7, 26.0) lymph nodes was detected, the median of lateral lymph nodes being 5.0 (2.0, 10.2). The median operation time was 254.5 (199.0, 325.2) minutes. The median intra-operative blood loss was 50.0 (30.0, 100.0) mL. All patients were diagnosed with adenocarcinoma by pathological examination of the operative specimen. Two patients developed postoperative intestinal obstruction, one lymphatic leakage, and one a perineal incision infection. There were no cases of anastomotic leakage. The median postoperative hospital stay was 6.0 (5.0, 7.0) days and the median follow-up time 23.5 (9.0, 36.7) months. During follow-up, three patients (7.1%) died of tumor recurrence and metastasis. Two (4.8%) experienced mild urinary dysfunction, and one (2.4%) had moderate postoperative erectile dysfunction. One patient (2.4%) was found to have prostate and lung metastases 3 month after surgery. The 3-year overall survival rate was 74.4%.Conclusions:Three sided encapsulation is a safe and feasible procedure for LLND, achieving accurate and complete clearance of lateral lymphatic tissue.
9.Feasibility of a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection for middle and low rectal cancer
Wenjuan LI ; Dechang DIAO ; Jiaxin LIN ; Jiahao WANG ; Weilin LIAO ; Xin TANG ; Jiaxin XIE ; Lin AO ; Xueyang ZHANG ; Xiaojiang YI ; Xiaochuang FENG ; Hongming LI ; Xinquan LU
Chinese Journal of Gastrointestinal Surgery 2023;26(10):968-976
Objective:To explore the feasibility and value of performing a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection (LLND) for middle and low rectal cancer.Methods:This was a retrospective review. The study cohort comprised patients who met the diagnostic criteria for rectal cancer according to the Chinese Guidelines for the Diagnosis and Treatment of Colorectal Cancer, had a short lymph node diameter of >5 mm on the lateral side within the 15 days before surgery, were evaluated as feasible candidates for laparoscopic total mesorectal excision+LLND surgery, had been diagnosed with low or intermediate level rectal cancer, and whose tumor was less than 8 cm away from the anal verge according to pathological examination of the operative specimen. Patients with a history of other malignant tumors of the abdomen or with incomplete follow-up data were excluded. Forty-two patients with middle and low rectal cancer who had undergone lateral lymph node dissection in diagnosis and treatment center of Gastrointestinal Cancer of Guangdong Hospital of Chinese Medicine from Jan.2018 to Dec.2022 were enrolled. There were 24 men (57.1%) and 18 women (42.9%) aged 58.4±11.8 years and the median BMI was 22.5 (19.3–24.1) kg/m 2. The main point of the three-sided encapsulation procedure is to expand the external side medial to the external iliac artery and vein, narrowing the range of exterior side dissection. The anterior-medial side is designed to expand the vesical fascia to define the range of anterior-medial side extension. The internal side is fully extended to the ureterohypogastric nerve fascia; the distal point of the caudal extension reaches the level of the Alcock canal and the bottom reaches the piriformis, enabling dissection of the obturator nerve and No.283 lymph nodes. No.263D lymph nodes are dissected by exposing the internal iliac artery and its branches, dissecting the group No.263P lymph nodes, and severing the inferior vesical artery. Finally, the lateral lymphatic tissue is completely resected. Relevant variables were recorded, including the number of lateral lymph nodes detected, the rate of lymph node metastasis, operation duration, intraoperative blood loss, postoperative complications, postoperative hospital stay, and 3-year overall survival rate. Results:Laparoscopic surgery was successfully completed in all patients with no conversions to open surgery and no intraoperative complications. Twenty-seven (64.3%) of the study patients underwent left-sided LLND, 10 (23.8%) right-sided LLND, and five (11.9%) bilateral LLND, with lymph nodes cleared on both sides. All patients' lymph nodes were examined pathologically. A median of 17.0 (11.7, 26.0) lymph nodes was detected, the median of lateral lymph nodes being 5.0 (2.0, 10.2). The median operation time was 254.5 (199.0, 325.2) minutes. The median intra-operative blood loss was 50.0 (30.0, 100.0) mL. All patients were diagnosed with adenocarcinoma by pathological examination of the operative specimen. Two patients developed postoperative intestinal obstruction, one lymphatic leakage, and one a perineal incision infection. There were no cases of anastomotic leakage. The median postoperative hospital stay was 6.0 (5.0, 7.0) days and the median follow-up time 23.5 (9.0, 36.7) months. During follow-up, three patients (7.1%) died of tumor recurrence and metastasis. Two (4.8%) experienced mild urinary dysfunction, and one (2.4%) had moderate postoperative erectile dysfunction. One patient (2.4%) was found to have prostate and lung metastases 3 month after surgery. The 3-year overall survival rate was 74.4%.Conclusions:Three sided encapsulation is a safe and feasible procedure for LLND, achieving accurate and complete clearance of lateral lymphatic tissue.
10.Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies.
Shenli ZHANG ; Qianru XU ; Jifei YANG ; Qingmei LI ; Yaning SUN ; Xueyang LI ; Yanan WANG ; Xiangxiang NIU ; Xiaotian QU ; Jinxuan CHEN ; Erqin ZHANG ; Gaiping ZHANG
Chinese Journal of Biotechnology 2022;38(5):1981-1993
The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
Animals
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Antibodies, Viral
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Chickens
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HN Protein/metabolism*
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Newcastle Disease/prevention & control*
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Newcastle disease virus/metabolism*
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Oryza/genetics*


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