1.Transcriptomic Analysis of Wuzi Yanzongwan on Testicular Spermatogenic Function in Semi-castrated Male Mice
Dixin ZOU ; Yueyang ZHANG ; Xuedan MENG ; Wei LU ; Shuang LYU ; Fanjun ZENG ; Kun CHEN ; Chang LIU ; Zhongxiu ZHANG ; Yu DUAN ; Yihang DAI ; Zhaoyi WANG ; Zhimin WANG ; Ruichao LIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):61-69
ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice, and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group, model group, testosterone propionate group(0.2 mg·kg-1·d-1, intramuscular injection) and Wuzi Yanzongwan group(1.56 g·kg-1·d-1, intragastric administration) according to body weight, with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function, while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention, hematoxylin-eosin(HE) staining was used to observe the histopathology of testis, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of serum testosterone(T), luteinizing hormone(LH) and follicle stimulating hormone(FSH). The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group, the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened, and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) to verify the transcriptome data. Through the annotation analysis of Gene Ontology(GO) and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG), the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group, the testicular tissue of mice in the model group showed spermatogenic injury, contraction and vacuolization of the seminiferous tubules, reduction of spermatogenic cells at all levels, widening of the interstitial space, obstruction of spermatogonial cell development and other morphological abnormalities, and serum T significantly decreased, LH significantly increased(P<0.01), and FSH elevated but no statistically significant difference, the count and vitality of epididymal sperm significantly decreased(P<0.01). There were 882 differentially expressed mRNAs in the testicular tissues, of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group, the damage to testicular tissue in the Wuzi Yanzongwan group was reduced, the structure of the seminiferous tubules was intact, vacuolization was reduced, and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased, LH significantly decreased(P<0.01), and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased(P<0.01). Moreover, Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice, of which 32 were up-regulated and 127 were down-regulated, and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis, renewal, differentiation, metabolism, apoptosis and signal transduction of spermatogenic cells, and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function, endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice, and its mechanism may be related to the regulation of testicular transcriptional regulatory network, the synthesis of sex hormones in testicular interstitial cells, the function of spermatogenic stem cells, the whole cell cycle process of spermatogenesis, as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.
2.The value of quantitative parameters of diffusion weighted imaging in differentiating the nature of ovarian epithelial tumors
Jiaojiao FAN ; Yang LIU ; Ruichao LIU ; Guanxiong LI ; Ziteng XIE ; Wei XIA
Journal of Practical Radiology 2024;40(9):1468-1471,1498
Objective To investigate the value of minimum apparent diffusion coefficient(ADCmin)and delta apparent diffusion coefficient(dADC)of quantitative parameters of MR diffusion weighted imaging(DWI)in differentiating benign from stage Ⅰ malignant ovarian epithelial tumors.Methods A total of 112 patients with primary ovarian epithelial tumors confirmed by operation and pathology(43 benign tumors and 69 stage Ⅰ malignant tumors)were retrospectively selected.All patients underwent routine MRI and DWI before operation.The ADCmin and dADC of solid portion were measured and compared between the two groups.Logistic regression anal-ysis and receiver operating characteristic(ROC)curve were used to evaluate diagnostic performance of each parameter alone and in combination in differentiating ovarian epithelial benign and stage Ⅰ malignant tumors.Results The ADCmin was greater in benign tumors than that in stage Ⅰ malignant tumors,while dADC was smaller in benign tumors than that in stage Ⅰ malignant tumors(P<0.05).Logis-tic regression analysis showed that ADCmin and dADC were independent risk factors for stage Ⅰ malignant tumors.With threshold of 1.183 × 10-3 mm2/s for ADCmin and 0.579 × 10-3 mm2/s for dADC,the area under the curve(AUC)was 0.802 and 0.857,sensitivity and specificity of 0.885,0.808,0.714,0.857,respectively.According to regression equation logit(P)=7.154+0.267 × dADC-1.016 X ADCmin,the AUC(0.951)was greater than that of single parameter,the sensitivity was 0.890,the specificity was 0.995.Conclusion ADCmin and dADC are useful for differentiating ovarian epithelial benign from stage Ⅰ malignant tumors,and ADCmin combined with dADC can improve the diagnose accuracy before operation.
3.Advances in the gene therapy of primary hyperoxaluria
Yukun LIU ; Ruichao ZHAN ; Yucheng GE ; Wenying WANG
Chinese Journal of Urology 2023;44(3):237-240
Primary hyperoxaluria (PH) is a rare autosomal recessive hereditary disease, characterized by calcium oxalate kidney stone and nephrocalcinosis caused by defects in enzymes of liver glyoxylate metabolism. Up to now, treatment options for PH are limited. Although medication treatment and liver transplantation can slow down the progression and mitigate the symptoms, the evidence for them turned out to be weak. In recent years, breakthroughs in biotechnology provide novel promising directions for drug development. Small interfering RNA drugs, such as lumasiran and nedosiran, selectively reduce hepatic expression of glycolate oxidase and lactate dehydrogenase respectively, reducing hepatic oxalate production and urinary oxalate levels in PH patients. Gene-editing, such as CRISPR/Cas9, will be a potential treatment method of PH. This review encompasses recent developments in the gene therapy of PH.
4.Renal abscess induced by fish bone ingestion: a case report
Yucheng GE ; Zhenqiang ZHAO ; Ruichao ZHAN ; Yukun LIU ; Chenglin ZHAO ; Tingting ZHANG ; Ye TIAN ; Wenying WANG
Chinese Journal of Urology 2023;44(9):704-705
Renal abscess caused by fish bone ingestion is extremely rare and has not been reported in the literature. A male patient presented with a 1-week history of flank pain and a 2-day history of fever. Urinary ultrasound and CT scan showed an irregular hypodense lesion in the left kidney and blurred thickening of the descending colon wall. Three-dimensional CT reconstruction images revealed a needle-like foreign body, which perforated from the descending colonic lumen to the left kidney. The patient had accidentally eaten fish bone one week prior. On the basis of clinical data, the diagnosis of renal abscess caused by foreign body was suspected. Accordingly, laparotomy was performed, the abscess was drained, and the colon was repaired. The foreign body was confirmed to be fish bone. The postoperative condition of the patient was uneventful, and the patient remained well in the 3 months' follow-up without any further complaints.
5.Advances of monogenic kidney stone diseases associated with purine metabolism
Yukun LIU ; Yucheng GE ; Ruichao ZHAN ; Wenying WANG
Chinese Journal of Urology 2022;43(2):156-160
Some kidney stones are caused by single gene mutations, and monogenic kidney stone diseases associated with purine metabolic disorder mainly including adenine phosphoribosyltransferase(APRT) deficiency, hypoxanthine-guanine phosphoribosyltransferase(HPRT)deficiency, hereditary xanthinuria(HX), and some diseases caused by gene mutations such as PRS1, SLC22A12, SLC2A9 and ABCG2. Such diseases can lead to abnormal metabolism of purine and uric acid, and then form 2, 8-dihydroxyadenine stones, uric acid stones or xanthine stones. This kind of diseases are rare, the genotype and phenotype of different types of monogenic diseases related to purine metabolism have their own characteristics and are not widely recognized. At present, the main treatment is medical therapy. Gene sequencing will make the diagnosis and find more disease-related genes or mutations. Gene editing, such as CRISPR/Cas9 technology, makes it possible to cure monogenic kidney stone diseases associated with purine metabolism disorder in the future.
6.Molecular pathology and clinical implications of diffuse glioma
Ruichao CHAI ; Shengyu FANG ; Bo PANG ; Yuqing LIU ; Yongzhi WANG ; Wei ZHANG ; Tao JIANG
Chinese Medical Journal 2022;135(24):2914-2925
The prognosis for diffusely infiltrating gliomas at World Health Organization (WHO) grade 2-4 remains dismal due to their heterogeneity. The rapid development of genome-wide molecular-profiling-associated studies has greatly promoted the accuracy of glioma classification. Thus, the latest version of the WHO classification of the central nervous system tumors published in 2021 has incorporated more molecular biomarkers together with histological features for the diagnosis of gliomas. Advanced usage of molecular pathology in clinical diagnostic practice provides also new opportunities for the therapy of patients with glioma, including surgery, radiotherapy and chemotherapy, targeted therapy, immunotherapy, and more precision clinical trials. Herein, we highlight the updates in the classification of gliomas according to the latest WHO guidelines and summarize the clinically relevant molecular markers by focusing on their applications in clinical practice. We also review the advances in molecular features of gliomas, which can facilitate the development of glioma therapies, thereby discussing the challenges and future directions of molecular pathology toward precision medicine for patients with glioma.
7.Summary of experience in the establishment of rat models with orthotopic liver transplantation from donation after cardiac death
Ruichao WU ; Zhaoyu HUANG ; Li ZHANG ; Junhan LIU ; Kepu ZHENG ; Jianghua RAN
Organ Transplantation 2018;9(4):304-310
Objective To explore the skills and summarize the experience in the establishment of orthotopic liver transplantation rat models from donation after cardiac death (DCD). Methods According to the time of warm ischemia, 120 rats were divided into 3 groups: group A (warm ischemia for 0 min, n=40 pairs), group B (warm ischemia for 10 min, n=40 pairs) and group C (warm ischemia for 20 min, n=40 pairs). Orthotopic liver transplantation was performed by the modified two-cuff technique in 3 groups. The time of each stage of surgery was recorded in 3 groups. The survival rate at the end of surgery, 24 h, 72 h and 7 d after surgery was recorded in 3 groups. The dead rats were immediately subject to anatomical examination to identify the cause of death. Results The cold ischemia time of donor liver, anhepatic phase and operation time of the recipients did not significantly differ among three groups (all P>0.05). In groups A, B and C, the survival rate at the end of surgery was 97%, 97%, and 100% respectively. The survival rate at postoperative 24 h was 92%, 90% and 92% respectively. The survival rate at postoperative 72 h was 90%, 80% and 77% respectively. The survival rate at postoperative 7 d was 85%, 70% and 57% respectively. The survival rate at the end of surgery, postoperative 24 h and 72 h did not significantly differ among 3 groups (all P>0.05). At postoperative 7 d, the survival rate in group C was significantly lower than that in group A (P<0.05). Surgical operation was the major cause of intraoperative and postoperative 24 h death. Bile leakage and ischemic hepatic failure were the causes of death at postoperative 72 h. Biliary duct complications were the main causes of death at postoperative 7 d. The quantity of rats developing with biliary duct complications was increased along with the prolongation of warm ischemic time. Conclusions The success of stable establishment of rat models with orthotopic liver transplantation from DCD depends upon the protection of the liver and biliary function. The difficulty lies in the anastomosis of the suprahepatic inferior vena cava and the shortening of anhepatic phase.
8.Prevention and management of pure-NOTES transanal total mesorectal excision for rectal cancer with postoperative anastomotic complications
Zhen RONG ; Gang YU ; Xiaoming WANG ; Ruichao GENG ; Xiangzhi WANG ; Longyi CHEN ; Fan YANG ; Xue LIU ; Yongjuan LI
International Journal of Surgery 2018;45(8):519-522,封3
Objective To investigate the the preventive and management methods of pure-NOTES transanal total mesorectal excision (pure-NOTES TaTME) with postoperative anastomotic complications.Methods Retrospectively analyzed the clinical data of 59 cases with low and middle rectal cancer who were underment pure-NOTES TaTME in Linzi District People's Hospital,and discussed the situction of the complications.Results Postoperative anastomotic complications were occurred in 3 cases,anastomotic leakage in 1 case,anastomotic stenosis in 1 case,anastomotic stenosis and leakage in 1 case,accounting for 5.1%.Conclusions For suitable rectal neoplasms patients,pure-NOTES TaTME operation doesn't increase the incidence of anastomotic complication,and it's is safe and feasible.Preoperative preparation,good blood supply,tension-free anastomosis,and correct choice and using of stapler and anastomotic drainage tube are the key to reduce anastomotic complications.
9.The study on behavioral characteristics of circadian rhythm disturbance in transgenic mice of Alzheimer's disease
Xiuli CAO ; Xuesong QU ; Ruichao LIU ; Jianming DING ; Meina WU
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(1):2-7
Objective To investigate the behavioral characteristics of circadian rhythm disturbance in 3xTg-AD mice.Methods The free-running period,average activity per hour,total amount of exercise and circadian amplitude were examined with voluntary wheel-running test in 3-to 9-month-old C57BL/6 and 3xTg-AD mice under 12 h light/12 h dark cycle and constant darkness environment.Results In constant darkness environment,the free-running period in 3-month-old 3xTg-AD mice was (23.2±0.4) h,and shorter than the period((23.5±0.2) h) in C57BL/6 mice.In 6-and 9-month-old 3xTg-AD mice,activity pattern was disorganized,without clear boundary between activity and rest phase.The free-running period was unavailable.The circadian amplitude and total exercise amount were(40.6± 11.5) counts/5 min and(2.6±0.1) × 104 counts/d,(37.0±20.8) counts/5 min and(2.3±0.4) × 104 counts/d,(29.3± 11.0) counts/5 min and(1.6± 0.9) × 104 counts/d in 3-,6-and 9-month-old 3xTg-AD mice,respectively,which was significantly lower than that in age-matched C57BL/6 mice.In 12 h light/12 h dark cycle,the circadian amplitude and total exercise amount of 3-,6-and 9-month-old 3xTg-AD mice were (87.0 ± 37.8) counts/5 min and (2.2 ± 0.8) × 104 counts/d,(25.9± 6.3) counts/5 min and (1.1 ± 0.2) × 104 counts/d,(14.3 ± 5.7) counts/5 min and (0.6 ± 0.3)× 104 counts/d respectively,and with a significant decrease from the age of 6 months.Meanwhile,the locomotor activity decreased at night and increased during the day.Conclusion The endogenous circadian rhythm disturbance emerges in 3xTg-AD mice at 3-month-old,while the exogenous circadian rhythm disorder appears at 6-month-old;the degree of disorder in circadian rhythm is gradually aggravated with the increase of age in of the AD mice.
10.Assessing the quality of rodent laboratory animals in Beijing area by pathological diagnosis in October 2014
Chao LI ; Haodi DONG ; Linkai XU ; Chunyu WANG ; Zhiqi SONG ; Chunfa LIU ; Chaosi LI ; Ruichao YUE ; Guangyu CHENG ; Huajia ZHAO ; Deming ZAO ; Xiaomin YIN ; Xiangmei ZHOU ; Lifeng YANG
Chinese Journal of Comparative Medicine 2015;(5):67-70
Objective To assess the health status of rodent laboratory animals by pathological diagnosis, our lab has being take apart in investigating the quality of laboratory animals in Beijing area for years and offer some advices for standardized breeding to ensure accurate results of scientific research.This paper focuses on the analysis of laboratory rodent samples that collected in October 2014.Methods We collected the heart, liver, spleen, lung, kidney, large intestine and small intestine, and put these organs into 10%Calcium formaldehyde solution for fixation, and then prepared into two different sections for optical microscopy observation including all paraffin specimens stained with H&E and the frozen sections stained with Oil Red-O and PAS.Results The vast majority of laboratory rodents were up to standard, but there still a problem in individual units.The main problem is liver and lung disease.The rate of Hepatocyte swellingis 6%(mouse), 2.5% (rat), 8.2% (guinea pig), moreover part of them were lipidosis, according to Oil Red-O stain.the mainly problem of lung is congestion ,edema and Interstitial pneumonia ,the detectable rate of pulmonarydiseases is 15.5%(guinea pig).Conclusions The vast majority of laboratory rodents were pathologically diagnosed as healthy animals.The liver disease may be caused by improper feeding.And disease of lung may led by haze, unqualified bedding and low temperature.

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