1.Study on the biological characteristics of human nucleus pulposus decellularized extracellular matrix scaffolds with different degrees of degeneration
Rong CHEN ; Bing RAN ; Ruidan ZHONG
Chinese Journal of Spine and Spinal Cord 2024;34(9):937-949
Objectives:To explore the biological characteristics of human nucleus pulposus decellularized extracellular matrix(NP-dECM)scaffolds with different degrees of degeneration(Ⅱ,Ⅲ,Ⅳ),and to select the NP-dECM scaffold with the best biological characteristics.Methods:Nucleus pulposus mesenchymal stem cells(NPMSCs)were isolated by collagenase type n digestion method.Cell adhesion,cell morphology and growth of NPMSCs were observed under microscope.The cell surface markers CD73,CD90,CD105,CD34,CD45,HLA-DR were detected by flow cytometry,and the differentiation ability of NPMSCs was observed by adipogenic,osteogenic and chondrogenic induction,so as to confirm that they were the seed cells needed for this study.Human NP-dECM scaffolds with different degrees of degeneration were prepared by physical,stain remover and nuclease(group Ⅱ-D,Ⅲ-D and Ⅳ-D).The decellular effect of NP-dECM scaffold was evaluat-ed by visual observation,quantitative and qualitative detection of DNA and extracellular matrix components,and SEM.The cytocompatibility of human NP-dECM scaffolds with different degrees of degeneration was e-valuated using CCK-8 method and living/dead cells in vitro.Results:Human NPMSCs isolated and cultured from the nucleus pulposus tissue grew adherent,and the cells were short spindle shaped and spindle-shaped.The third passage of human NPMSCs expressed high levels of surface markers such as CD73,CD90 and CD105,and low levels of surface markers such as CD34,CD45 and HLA-DR.It met the criteria of mes-enchymal stem cells.Human NP-dECM scaffolds with three degrees of degeneration remained intact,and the scaffolds of each group were loose and porous under electron microscope.HE staining,DAPI staining and DNA determination showed that the NP-dECM scaffold effectively removed cell components in the three de-grees of degeneration.The quantitative and qualitative detection results of GAG and HYP showed that ECM was well retained,and the GAG retention was in the order of group Ⅱ-D>group Ⅲ-D>group Ⅳ-D from high to low.There was no statistical significance in HYP content between groups Ⅱ-D and Ⅲ-D,which was higher than that in group Ⅳ-D.The results of CCK-8 and live/dead cell staining showed that groups Ⅱ-D,Ⅲ-D and Ⅳ-D showed good cytocompatibility.Conclusions:The Ⅱ degree degeneration of human nucleus pulposus tissue is the best choice for the preparation of human NP-dECM scaffold.
2.Exploration of cross-cultivar group characteristics of a new cultivar of Prunus mume 'Zhizhang Guhong Chongcui'.
Xiaotian QIN ; Mengge GUO ; Shaohua QIN ; Ruidan CHEN
Chinese Journal of Biotechnology 2024;40(1):239-251
'Zhizhang Guhong Chongcui' is a new cultivar of Prunus mume with cross-cultivar group characteristics. It has typical characteristics of cinnabar purple cultivar group and green calyx cultivar group. It has green calyx, white flower, and light purple xylem, but the mechanism remains unclear. In order to clarify the causes of its cross-cultivar group traits, the color phenotype, anthocyanin content and the expression levels of genes related to anthocyanin synthesis pathway of 'Zhizhang Guhong Chongcui', 'Yuxi Zhusha' and 'Yuxi Bian Lü'e' were determined. It was found that the red degree of petals, sepals and fresh xylem in branches was positively correlated with the total anthocyanin content. MYBɑ1, MYB1, and bHLH3 were the key transcription factor genes that affected the redness of the three cultivars of flowers and xylem. The transcription factors further promoted the high expression of structural genes F3'H, DFR, ANS and UFGT, thereby promoting the production of red traits. Combined with phenotype, anthocyanin content and qRT-PCR results, it was speculated that the white color of petals of 'Zhizhang Guhong Chongcui' were derived from the high expression of FLS, F3'5'H, LAR and ANR genes in other branches of cyanidin synthesis pathway, and the low expression of GST gene. The green color of sepals might be originated from the relatively low expression of F3'H, DFR and ANS genes. The red color of xylem might be derived from the high expression of ANS and UFGT genes. This study made a preliminary explanation for the characteristics of the cross-cultivar group of 'Zhizhang Guhong Chongcui', and provided a reference for molecular breeding of flower color and xylem color of Prunus mume.
Animals
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Anthocyanins
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DNA Shuffling
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Flowers/genetics*
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Porifera
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Prunus/genetics*
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Glutamine/analogs & derivatives*
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Plant Extracts
3.The dynamic expression and internal relationship of matrix metalloproteinase-9, interleukin-1β and tissue inhibitor of metalloproteinase-1 in hypoxic-ischemic injury of neonatal rats
Na LI ; Chunying LIU ; Yang WANG ; Ruidan CHEN ; Xiaofeng LI
Chinese Pediatric Emergency Medicine 2021;28(1):45-49
Objective:To investigate the expression levels and internal relations of matrix metalloproteinase-9(MMP-9), interleukin(IL)-1β and tissue inhibitor of metalloproteinase-1(TIMP-1) within 24 h after cerebral hypoxia and ischemia(HI) in neonatal rats.Methods:SD rats aged 7 days were randomly divided into HI group( n=40), and sham operation group(sham group, n=40). The HI models were established and divided into 5 subgroups with those at 8 cases in each group at 5 postoperative time points(0, 4, 8, 12, 24 h). Pathological changes of cerebral cortex were observed by HE staining, and the expressions of MMP-9, IL-1β and TIMP-1 were detected by Western blot and RT-PCR.In addition, the hypoxia models of mouse hippocampal neuron HT22 cells were established.With adding MMP-9 inhibitor and MMP-9 agonist, the relations among MMP-9, IL-1β and TIMP-1 were further confirmed. Results:Slight nuclear abnormalities and cell body swelling occurred in cerebral cortex at 4 h after HI, and neuronal eosinophilic changes were most obvious at 12 h. Compared with those at 8 h and 12 h, the expressions of MMP-9 and IL-1β at protein and mRNA level significantly increased at 24 h after HI( P<0.01). While MMP-9 inhibitor was added to HT22 cells under hypoxia at 4 h, IL-1β mRNA expression was down-regulated. Conclusion:In the early stage of hypoxic ischemic encephalopathy in newborn rats, the expressions of MMP-9 and TIMP-1 are time-dependent.Inhibition of MMP-9 expression could reduce the level of Interleukin-1β.
4.Energy metabolism characteristic with risk of secondary bacterial infection in patients with hepatitis B virus-related chronic liver disease
Shenglong LIN ; Minghua LIN ; Xiangmei WANG ; Xiumin CHEN ; Hanhui YE ; Huaxi MA ; Dongqing ZHANG ; Wenjun WU ; Jiahuang LIN ; Ziyuan LIAO ; Ruidan ZHENG ; Haibing GAO
Chinese Journal of Hepatology 2021;29(6):558-564
Objective:To investigate and analyze the energy metabolism characteristics and the correlation between energy metabolism and the risk of secondary bacterial infection in patients with hepatitis B virus-related chronic liver disease (HBV-CLD).Methods:Data of 183 cases admitted to the Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2020 were retrospectively analyzed. 79 cases of chronic hepatitis B, 51 cases of hepatitis B-related liver cirrhosis, and 53 cases of hepatitis B-related liver failure were collected. Among them patients with liver failure and decompensated liver cirrhosis were defined as severe liver disease group. The Quark RMR indirect calorimetry (COSMED Corporation, Italy) was used to exam the patients' energy metabolism condition, and the incidences of secondary bacterial infection of the patients during hospitalization were recorded. Shapiro-Wilk test and normal QQ plot were used to analyze the normal distribution of continuous variable data, which was consistent with the normal distribution and was described by mean ± standard deviation. In addition, if it did not conform to the normal distribution, the median and interquartile distance were used to describe it. Levene’s test was used to test the homogeneity of variance of the data, which was consistent with the normal distribution. The t-test was used to compare the means of the two groups of samples. One-way analysis of variance was used to compare the mean values of the three groups of samples, and then the Tukey's test was used to compare the two groups. If the variance was uneven or did not conform to the normal distribution, the Wilcoxon rank sum test was used to compare the differences between the two groups. Kruskal-Wallis test (H test) was used to compare the differences between the three groups of samples, and then the Dunnett’s test (Z test) was used for comparison between the two groups. Categorical variable data were analyzed using chi-square test. Logistic regression analysis was used to screen independent risk factors, and the criteria for variable inclusion ( P < 0.05). Results:The respiratory entropy (RQ) and non-protein respiratory entropy (npRQ) of the three groups had statistically significant difference ( P < 0.05). Among them, the RQ and npRQ of the chronic hepatitis B group were higher than hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group. There were statistically significant differences in fat oxidation rate (FAT%) and carbohydrate oxidation rate (CHO%) between the three groups ( P < 0.05). Compared with hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group, chronic hepatitis B group ( P < 0.05) had lower FAT% and higher CHO%. There were no statistically significant differences in the measured and predicted resting energy expenditure and protein oxidation rate (PRO%) between the three groups. The incidence of secondary bacterial infection in patients with severe liver disease was 48.39% (45/93). Compared with the non-infected group, the RQ and npRQ values ??of the infected group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Logistic regression analysis showed that glutamyltransferase, cholesterol, and npRQ were independent risk factors for secondary bacterial infections in patients with severe liver disease. Glutamyltransferase elevation, and cholesterol and npRQ depletion had suggested an increased risk of secondary bacterial infection. Subgroup analysis of patients with hepatitis B-related liver failure also showed that compared with non-infected group, RQ value and npRQ value of secondary bacterial infection group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Conclusion:Patients with hepatitis B virus-related chronic liver disease generally have abnormal energy metabolism. Low RQ, npRQ, CHO% and high FAT% are related to the severity of the disease; while npRQ reduction is related to the risk of secondary bacterial infection in patients with severe liver disease, and thus can be used as a clinical prognostic indicator.
5.Analysis of information about coronavirus disease 2019 on WeChat official accounts of CDCs in Zhejiang Province
YAN Xiaotong ; LAI Ruidan ; REN Shaofan ; WU Chao ; XIE Jun ; CHEN Xifan ; XU Jinhang
Journal of Preventive Medicine 2020;32(3):226-229
Objective:
To analyze the information about coronavirus disease 2019 (COVID-19) on WeChat official accounts of centers for disease control and prevention (CDCs) in Zhejiang Province from January 20 to February 5,2020,so as to provide reference for improving the effects of health communication by WeChat official accounts.
Methods:
The number,content and pageview of the information about COVID-19 on WeChat official accounts of one provincial and eleven municipal CDCs from January 20 to February 5 were collected and analyzed. The number of new followers and WeChat communication power index (WCI) were employed to evaluate the communication effect.
Results:
By February 5,those WeChat public official accounts pushed 629 pieces of information about COVID-19. The pageviews were 3 713 428 in total and 5 903.70 on average. There were totally 633 008 followers,including 110 341 new followers which contributed to a growth rate of 21.11%. The average WCI was 677.81. The WCIs of eight official accounts were higher than 500,with “Zhejiang Health Education” the highest (1 021.95). The daily pageviews peaked on January 20,21,25 and 31. Among the top 15 pieces of information in pageviews,there were 7 pieces for epidemic announcements,3 pieces for popular science and 5 pieces for behavioral intervention.
Conclusions
The WeChat official accounts of CDCs in Zhejiang Province pushed the information about COVID-19 in line with the progress of the epidemic and the demand of the public,leading to a higher attention and better communication effect.
6.Clinical research advances in traditional Chinese medicine in treatment of icteric hepatitis
Qiyu GUO ; Jianneng CHEN ; Ruidan ZHENG
Journal of Clinical Hepatology 2017;33(5):833-837
The pathogenesis of icteric hepatitis is that pathogenic dampness blocks the middle energizer or blood stasis blocks the bile duct,so that the bile is not excreted normally and moves to the skin surface,which makes the skin yellow.Traditior al Chinese medicine (TCM) has unique advantages in the prevention and treatment of icteric hepatitis.This article systematically introduces the etiology and pathogenesis of icteric hepatitis and the TCM syndrome differentiation therapy for icteric hepatitis,in order to provide a reference for the clinical treatment of icteric hepatitis and the improvement in its prognosis.
7.Clinical analysis of enteral nutrition in 47 children
Ruidan ZHUANG ; Lujing TANG ; Youhong FANG ; Kerong PENG ; Jie CHEN
Chinese Journal of Pediatrics 2016;54(7):500-503
Objective To explore the efficacy and safety of the application of enteral nutrition (EN) in gastrointestinal disease in children,and to explore the possibility of the implementation of family EN.Method Retrospective analysis of disease spectrum,EN approach,preparation,speed and time as well as adverse reactions and outcomes in 47 pediatric patients with gastrointestinal disease underwent EN therapy during July 2014 to March 2015.The nutrition indicators before and after EN therapy were compared by paired t-test.Result A total of 47 patients were selected,27 male (57%) and 20 female (43%),aged 0.8 (0.3,4.0) years,9 with mechanical or chemical damage to the esophagus,7 with inflammatory bowel disease (including ulcerative colitis and Crohn's disease),6 with chronic diarrhea,5 with acute pancreatitis,3 with acute diarrhea and severe malnutrition,3 with short bowel syndrome,3 with improper feeding,3 with feeding difficulties,3 with protein losing enteropathy,2 with post-enterostomy,2 with enterocolitis,1 with gastroesophageal reflux,were diagnosed.Of 47 cases,22 were given oral nutrition,28 were fed with nasogastric tube and 4 with nasojejunal tube feeding,2 with percutaneous endoscopic gastrojejunostomy tube feeding for each.In these tube-feeding cases,20 cases were treated with continuous infusion and 21 cases with intermittent infusion.Eleven cases were fed with amino acid formula;21 cases took the choice of peptide formulations;16 cases chose whole protein formula,including six cases who chose 3.3-4.2 kJ/ml higher energy density formula,10 cases selected common energy density formula including breast milk.Twenty-one cases suffered from different degrees of adverse reactions,including vomiting in 7 cases,abdominal pain and bloating in 3,diarrhea in 12,secondary respiratory infections in 5.Five patients were discharged after giving up of treatment by parents due to poor efficacy on primary disease;3 cases were transferred to other departments for further treatment;15 cases were discharged with a feeding tube for family nutrition and specialist out-patient treatment.The rest 24 cases were all improved and discharged.There were significant differences in nutrition indicators before and after EN,weight-for-age Z score (WAZ)(-2.3 ± 1.9 vs.-1.9 ± 1.8,t =4.156,P =0.000),weight-for-height Z score (WHZ) (-1.9 ± 1.7 vs.-1.2±1.5,t=3.714,P=0.001),albumin ((35 ±9)g/L vs.(39 ±6) g/L,t=3.017,P=0.005) and prealbumin ((0.11 ±0.05)g/L vs.(0.18 ±0.07)g/L,t=5.144,P=0.000).Conclusion EN is suitable for a variety of children's digestive diseases,which can improve the nutritional status of the patients and was safe for clinical application.As the implementation of EN is simple and has good compliance,family EN is proven to be feasible.
8.A multi-center clinical study of a novel controlled attenuation parameter for assessment of fatty liver.
Feng SHEN ; Ruidan ZHENG ; Yuqiang MI ; Junping SHI ; Guofeng CHEN ; Jianneng CHEN ; Liang XU ; Qin PAN ; Leiming XU ; Jiangao FAN
Chinese Journal of Hepatology 2014;22(12):926-931
OBJECTIVETo evaluate the controlled attenuation parameter (CAP) assessment of fatty liver and choose a cut-off value of hepatic steatosis more than 5%.
METHODSConsecutive patients, 18 years or older, who had undergone percutaneous liver biopsy and CAP measurement were recruited from five liver healthcare centers in China. All enrollees were categorized as hepatic steatosis grade S0 (<5%) or S1 (5%). An M-probe equipped FibroScan 502 was used to capture CAP values. Receiver operating characteristic (ROC) curves were plotted, and the areas under (AU) the curves were calculated to determine the diagnostic efficacy. The CAP cut-off values at the optimal thresholds were defined by maximum Youden indices; sensitivity and specificity were also calculated.
RESULTSA total of 332 patients were enrolled in the study, including 67 patients with non-alcoholic fatty liver disease (NAFLD) and 265 with chronic hepatitis B (CHB) viru: infection. The median age (inter quartile range, IQR) of the study cohort was 39.0 (32.0-50.5) years-old. There were 46 males (68.7%) in the NAFLD group, with a median age of 37.0 (28.0-45.0) years-old, and 182 males (68.7%) in the CHB group; the differences between the two groups in median age and male: female ratio did not reach statistical significance. Multivariate linear regression analysis identified steatosis grade and body mass index (BMI) as independently associated with CAP. The median (IQR) CAP values among patients with S0 and S1 grade steatosis were 215.0 (190.0-241.0) dB/m and 294.0 (255.0-325.5) dB/m (P<0.001), respectively. For all patients, when BMI was <25 kg/m2, the ability of the AUROC of the CAP to discriminate hepatic steatosis more than or equal to 5% was 0.853, and the optimal cut-off value was 244.5 dB/m; however, when BMI≥25 kg/m2, the AUROC was 0.835 and the optimal cut-off value 269.5 dB/m.
CONCLUSIONCAP can identify hepatic steatosis more than or equal to 5% and is applicable for the diagnosis of fatty liver if it is adjusted for BMI.
Adult ; Area Under Curve ; Bile ; Biopsy ; Body Mass Index ; China ; Fatty Liver ; Female ; Hepatitis B, Chronic ; Humans ; Linear Models ; Male ; Middle Aged ; Multivariate Analysis ; ROC Curve ; Tissue Extracts
9.Population ecology of medical shellfish and the infection rate of Angiostrongylus cantonensis in Longhai City,Fujian Province, China
Guohua LIN ; Mingsong HUANG ; Youzhu CHENG ; Ruidan ZHENG ; Shaohong CHEN ; Cuilan YAN ; Yanyan FANG ; Shaoxing LIN ; Yaoxiong ZHOU
Chinese Journal of Zoonoses 2014;(8):821-827
The aim of the study was to investigate the population ecology of medical shellfish and the infection of An-giostrongylus cantonensis in Longhai ,Fujian Province ,China .Aquatic and terrestrial shellfish were collected in survey points according to different types of breeding grounds .Then ,lung-microscopy method was involved in the detection of the lung tis-sue in Ampullaria gigas .Other shellfishes were mashed to detect the third-stage larvae of Angiostrongylus cantonensis .Hom-ogenization and lung microscopy were compared in the detection of the larvae of A .cantonensis in Achatina snails .Factors re-lated to the environment and influence of shellfish hosts were also included .Results showed that 8 species of molluscans were found ,including Pila gigas ,Bellamya aeruginosa ,Bellamya lithophaga ,Melanoides tuberculata ,Achatina fulica ,Vag-inulus alte ,Philomycus bilineatus ,and Bradybaenasimilaris with 1 673 specimens in 27 survey points from 9 townships .The infectionratewas19.78% inaverage.TheinfectionrateinV.altewas56.63% (47/83);theinfectionratesforA.fulicaand P .gigas were 39 .32% (92/234) and 27 .14% (130/234) ,respectively .The infection rate of each survey point was closely re-lated to the distances from the residents living area .Morever ,A .cantonensis larvae were detected in M .tuberculata .Lung mi-croscopy and homogenization method detection rate was 87 .1%and 100 .0% ,respectively .The difference was statistically sig-nificant .In conclusion ,V .alte ,A . fulica and P .gigas were A . cantonensist infection dominant population . The infection rate was closely related to micro-ecological environment for all kinds of shellfish .M .tuberculata was the new host of A .can-tonensis .Lung microscopy method should not be used in the qualitative screening detection of A . f ulica infected with A .can-tonensist .
10.Clinical characteristics of chronic hepatitis B patients with metabolic syndrome
Jianneng CHEN ; Ruidan ZHENG ; Jiangao FAN ; Qin PAN ; Bifen CHEN ; Feng SHEN
Chinese Journal of Infectious Diseases 2014;32(5):289-292
Objective To explore the clinical characteristics of chronic hepatitis B patients with metabolic syndrome (MS).Methods A total of 127 patients with untreated chronic hepatitis B infection were recruited.The patients were divided into two groups according to the diagnosis of MS,with 45 in MS group and 82 in non-MS group.Age,gender,body mass index (BMI),waist-to-hip ratio (WHR),fasting blood glucose (FBG),triglyceride (TG),total cholesterol (TC),aspartate aminotransferase (AST),alanine aminotransferase (ALT),hepatitis B e antigen (HBeAg),hepatitis B virus deoxyribonucleic acid (HBV DNA) were compared between groups.Liver biopsy was performed in 94 patients,including 31 in MS group and 63 in non-MS group,to compare the histological characteristics of liver between two groups.A chi-square test and t test were used for the data analysis.Results BMI,WHR,TG,TC,and FBG results of patients in MS group and nonMS group were (28.88±3.00) kg/m2,0.93±0.03,(2.77±0.17) mmol/L,(6.51±0.95) mmol/L,(6.67±0.45) mmol/L,and (24.64± 2.21) kg/m2,0.91±0.04,(1.50±0.65) mmol/L,(4.38±0.71) mmol/L,(4.91±0.92) mmol/L,respectively.Patients in MS group had higher BMI,WHR,FBG,TG and TC compared with those in non-MS group (t=9.109,3.245,2.642,3.762 and 2.586,respectively; all P<0.05).No statistical significant differences were found in age,ALT or AST between two groups (t=2.224,0.703 and 0.141,respectively; all P>0.05).Neither any statistical difference was found in gender,the positive rate of HBeAg and HBV DNA between two groups (x2 =1.662,0.037 and 0.944,respectively; all P>0.05).In inflammation activity comparison,the result of liver biopsy showed that 48.39 % (15/31) were classified as G0-G1 and 51.61%(16/31) as G2-G4 in MS group,and those in non MS group were 49.21%(31/63) and 50.79%(32/63),respectively.No statistical significance was reached (x2 =0.006,P>0.05).In fibrosis stage comparison,patients in MS group classifies as S0-S1 and S2-S4 were 32.26% (10/31) and 67.74%(21/31),respectively,and those in non-MS group were 60.32% (38/63)and 39.68% (25/63),respectively.The liver fibrosis in MS group was significantly more severe than that in non-MS group (x2 =6.546,P<0.05).Conclusions The CHB patients with MS have higher BMI,WHR,FBG,TG and TC.The presence of MS may promote the progress of liver fibrosis in CHB patients.


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