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1.
Chinese Pharmacological Bulletin ; (12): 308-316, 2024.
Article in Chinese | WPRIM | ID: wpr-1013586

ABSTRACT

Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.

2.
Journal of Southern Medical University ; (12): 225-231, 2023.
Article in Chinese | WPRIM | ID: wpr-971519

ABSTRACT

OBJECTIVE@#To explore the association between depressive symptoms and the risks of rapid decline in renal function and chronic kidney disease (CKD) in middle-aged and elderly with normal kidney function.@*METHODS@#The residents aged 40- 75 years with eGFR≥60 mL·min-1·1.73 m-2 without proteinuria in Lanzhou region, who participated in the "REACTION" study carried out in 2011, were selected and followed up in 2014. A total of 4961 individuals with complete and qualified data from the two surveys were included in the subsequent analysis. Based on PHQ-9 questionnaire scores, the baseline population was divided into two groups with and without depressive symptoms. Cox proportional hazard analysis was used to compare the incidences of rapid renal function decline and CKD between the two groups and study the association of depressive symptoms with the risk of these renal conditions.@*RESULTS@#PHQ-9 questionnaire scores were not found to correlate with baseline SCr, ALB, UACR or eGFR levels among the participarts (P>0.05). After a mean follow-up time of 3.4±0.6 years, 33.9% of the participants with depressive symptoms at baseline experienced a rapid decline in renal function and 3.6% progressed to CKD. During the follow-up, the incidence of rapid decline in renal function and the risk of developing CKD were not found to correlate with depressive symptoms in these participants (P>0.05) regardless of the type of the depressive syndromes.@*CONCLUSION@#Depressive symptoms are not associated with the risks of rapid renal function decline or progression to CKD in middle-aged and elderly with normal kidney function.


Subject(s)
Aged , Middle Aged , Humans , Cohort Studies , Depression , Glomerular Filtration Rate , Disease Progression , Renal Insufficiency, Chronic/epidemiology , Kidney/physiology , Risk Factors
3.
Chinese Journal of Surgery ; (12): 761-768, 2023.
Article in Chinese | WPRIM | ID: wpr-985820

ABSTRACT

Objectives: To analyze the influencing factors of No. 253 lymph node metastasis in descending colon cancer, sigmoid colon cancer, and rectal cancer, and to investigate the prognosis of No. 253 lymph node-positive patients by propensity score matching analysis. Methods: A retrospective analysis was performed on clinical data from patients with descending colon cancer, sigmoid colon cancer, rectosigmoid junction cancer, and rectal cancer who underwent surgery between January 2015 and December 2019 from the Cancer Hospital of the Chinese Academy of Medical Sciences, China-Japan Friendship Hospital, Peking Union Medical College Hospital, General Hospital of the Chinese People's Liberation Army, and Peking University Cancer Hospital. A total of 3 016 patients were included according to inclusion and exclusion criteria, comprising 1 848 males and 1 168 females, with 1 675 patients aged≥60 years and 1 341 patients aged<60 years. Clinical and pathological factors from single center data were subjected to univariate analysis to determine influencing factors of No. 253 lymph node metastasis, using a binary Logistic regression model. Based on the results of the multivariate analysis, a nomogram was constructed. External validation was performed using data from other multicenter sources, evaluating the effectiveness through the area under the receiver operating characteristic curve and the calibration curve. Using data from a single center, the No. 253 lymph node-positive group was matched with the negative group in a 1∶2 ratio (caliper value=0.05). Survival analysis was performed using the Kaplan-Meier method and Log-rank test. The Cox proportional hazards model was used to determine independent prognostic factors. Results: (1) The tumor diameter≥5 cm (OR=4.496,95%CI:1.344 to 15.035, P=0.015) T stage (T4 vs. T1: OR=11.284, 95%CI:7.122 to 15.646, P<0.01), N stage (N2 vs. N0: OR=60.554, 95%CI:7.813 to 469.055, P=0.043), tumor differentiation (moderate vs. well differentiated: OR=1.044, 95%CI:1.009 to 1.203, P=0.044; poor vs. well differentiated: OR=1.013, 95%CI:1.002 to 1.081, P=0.013), tumor location (sigmoid colon vs. descending colon: OR=9.307, 95%CI:2.236 to 38.740, P=0.002), pathological type (mucinous adenocarcinoma vs. adenocarcinoma: OR=79.923, 95%CI:15.113 to 422.654, P<0.01; signet ring cell carcinoma vs. adenocarcinoma: OR=27.309, 95%CI:4.191 to 177.944, P<0.01), and positive vascular invasion (OR=3.490, 95%CI:1.033 to 11.793, P=0.044) were independent influencing factors of No. 253 lymph node metastasis. (2) The area under the curve of the nomogram prediction model was 0.912 (95%CI: 0.869 to 0.955) for the training set and 0.921 (95%CI: 0.903 to 0.937) for the external validation set. The calibration curve demonstrated good consistency between the predicted outcomes and the actual observations. (3) After propensity score matching, the No. 253 lymph node-negative group did not reach the median overall survival time, while the positive group had a median overall survival of 20 months. The 1-, 3- and 5-year overall survival rates were 83.9%, 61.3% and 51.6% in the negative group, and 63.2%, 36.8% and 15.8% in the positive group, respectively. Multivariate Cox analysis revealed that the T4 stage (HR=3.067, 95%CI: 2.357 to 3.990, P<0.01), the N2 stage (HR=1.221, 95%CI: 0.979 to 1.523, P=0.043), and No. 253 lymph node positivity (HR=2.902, 95%CI:1.987 to 4.237, P<0.01) were independent adverse prognostic factors. Conclusions: Tumor diameter ≥5 cm, T4 stage, N2 stage, tumor location in the sigmoid colon, adverse pathological type, poor differentiation, and vascular invasion are influencing factors of No. 253 lymph node metastasis. No. 253 lymph node positivity indicates a poorer prognosis. Therefore, strict dissection for No. 253 lymph node should be performed for colorectal cancer patients with these high-risk factors.

4.
Chinese Journal of Surgery ; (12): 753-760, 2023.
Article in Chinese | WPRIM | ID: wpr-985819

ABSTRACT

Objective: To examine a predictive model that incorporating high risk pathological factors for the prognosis of stage Ⅰ to Ⅲ colon cancer. Methods: This study retrospectively collected clinicopathological information and survival outcomes of stage Ⅰ~Ⅲ colon cancer patients who underwent curative surgery in 7 tertiary hospitals in China from January 1, 2016 to December 31, 2017. A total of 1 650 patients were enrolled, aged (M(IQR)) 62 (18)years (range: 14 to 100). There were 963 males and 687 females. The median follow-up period was 51 months. The Cox proportional hazardous regression model was utilized to select high-risk pathological factors, establish the nomogram and scoring system. The Bootstrap resampling method was utilized for internal validation of the model, the concordance index (C-index) was used to assess discrimination and calibration curves were presented to assess model calibration. The Kaplan-Meier method was used to plot survival curves after risk grouping, and Cox regression was used to compare disease-free survival between subgroups. Results: Age (HR=1.020, 95%CI: 1.008 to 1.033,P=0.001), T stage (T3:HR=1.995,95%CI:1.062 to 3.750,P=0.032;T4:HR=4.196, 95%CI: 2.188 to 8.045, P<0.01), N stage (N1: HR=1.834, 95%CI: 1.307 to 2.574, P<0.01; N2: HR=3.970, 95%CI: 2.724 to 5.787, P<0.01) and number of lymph nodes examined (≥36: HR=0.438, 95%CI: 0.242 to 0.790, P=0.006) were independently associated with disease-free survival. The C-index of the scoring model (model 1) based on age, T stage, N stage, and dichotomous variables of the lymph nodes examined (<12 and ≥12) was 0.723, and the C-index of the scoring model (model 2) based on age, T stage, N stage, and multi-categorical variables of the lymph nodes examined (<12, 12 to <24, 24 to <36, and ≥36) was 0.726. A scoring system was established based on age, T stage, N stage, and multi-categorical variables of lymph nodes examined, the 3-year DFS of the low-risk (≤1), middle-risk (2 to 4) and high-risk (≥5) group were 96.3%(n=711), 89.0%(n=626) and 71.4%(n=313), respectively. Statistically significant difference was observed among groups (P<0.01). Conclusions: The number of lymph nodes examined was an independent prognostic factor for disease-free survival after curative surgery in patients with stage Ⅰ to Ⅲ colon cancer. Incorporating the number of lymph nodes examined as a multi-categorical variable into the T and N staging system could improve prognostic predictive validity.

5.
Acta Pharmaceutica Sinica ; (12): 3675-3685, 2022.
Article in Chinese | WPRIM | ID: wpr-964317

ABSTRACT

2-Oxoglutarate/Fe(II)-dependent dioxygenases (2-ODD) play an important role in plant primary and secondary metabolism. Based on the high-throughput sequencing platform Illumina NovaSeq 6000, the transcriptome of Salvia apiana Jepson was sequenced, and the obtained reads were de novo assembled. A total of 38 534 unigenes were obtained from the transcriptome. The assembled unigenes were annotated and 29 982 unigenes were given functional annotations. The 2-ODD genes were identified from the assembled S. apiana transcriptome database by bioinformatics methods, and the genes were analyzed, including the homology of the sequences, physicochemical characteristics, signal peptides, transmembrane domains, subcellular localization, secondary structure and tertiary structure, etc. The evolutionary relationships and the expression patterns of the identified 2-ODD genes were also analyzed. 39 full-length 2-ODD genes were identified from the transcriptome of S. apiana. The average length of these 2-ODD encoding proteins was 320 amino acids, the molecular weight was about 36.00 kDa, and most of them were hydrophilic proteins. Phylogenetic analysis divided these 2-ODD genes into several subfamilies. Gene expression analysis indicated that the 2-ODD genes were expressed in different parts of S. apiana, and the expression level of most genes was much higher in roots than that in leaves. This study can lay a foundation for further study of 2-ODD genes in S. apiana.

6.
Chinese Pharmacological Bulletin ; (12): 1579-1585, 2022.
Article in Chinese | WPRIM | ID: wpr-1014236

ABSTRACT

Aim To investigate whether and how Huy- izhenbao tablets regulated osteoarthritis pain.Methods We transected the meniscotibial ligament of mice, which caused osteoarthritis by destabilizing the medial meniscus ( DMM).Different doses of Huyizhenbao tablets (12.5,25,50 mg • kg-1) were administered intragastrically.Dynamic and static mechanical allo- dynia were measured.The spinal cord slices were pre¬pared to record miniature excitatory postsynaptic cur-rents (niEPSCs) and miniature inhibitory postsynaptic currents (mlPSCs) by using patch clamp electrophysi¬ological recordings.The phosphorylation of NMDA re¬ceptor ( N-methyl-D-aspartate receptors) (rluNl sub- unit at S897 residue ( pS897-GluNl ) was observed by immunohistochemistry.Results Huyizhenbao tablets dose-dependently attenuated the dynamic and static mechanical allodynia induced by DMM, reduced the frequency of niEPSCs and inhibited the pS897-GluNI level.Huyizhenbao tablets had no effects on mlPSCs.Conclusions Huyizhenbao tablets effectively alleviate osteoarthritis pain by blocking the presynaptic release of excitatory transmitter glutamate and inhibiting the phosphorylation of NMDA receptor in spinal cord dorsal horn.

7.
Chinese Pharmacological Bulletin ; (12): 387-393, 2022.
Article in Chinese | WPRIM | ID: wpr-1014138

ABSTRACT

Aim To discuss the effect of berberine ( BE) on the activity of HSV-1 virus infected HEp-2 cells and its related molecular mechanisms.Methods Hie infected cell model was constructed and divided into control group, infection group, low concentration group ( 5 (xmol • L 1 -BE) , medium concentration group ( 10 (xmol • L '-BE) and high concentration group ( 15 (xmol • L '-BE) ) , and then incubated for 24 hours.qRT-PCR was used to determine HSV-1 infection-related genes ( gD, ICP-4, ICP-8, ICP-27 ) and mRNA expression levels of LncBNA NRAV, miR- 299-3p, RAB5C.CCK-8 method and flow cytometry were applied to detect cell viability and apoptotic rate.The expression levels of PI3K/AKT signaling pathway and JNK/p38 MAPK signaling pathway related protein were analysed by WB.Results It was found that BE j j reduced the mRNA expression of gD, ICP-4, ICP-8, anrl ICP-27, improved cell viability, and inhibited eell apoptosis.BE promoted the expression of miR-299-3p by inhibiting LncRNA NRAV and RAB5C.BE inhibited the protein expression levels of PBK/AKT signaling pathway and JNK/p38 MAPK signaling pathway proteins PI3K, AKT, JNK, and P38.Conclusions The mechanism that BE enhances the activity of HEp-2 cells after HSV-1 infection and suppresses its apoptosis may be related to LncRNA NRAV and RAB5C targeting competitive binding to miH-299-3p, inhibiting the activation of PBK/AKT signaling pathway and JNK/ p38 MAPK signaling pathway.

8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 92-99, 2022.
Article in Chinese | WPRIM | ID: wpr-940293

ABSTRACT

Malignant tumor is a serious threat to human life and health. The prevalence and mortality of malignancies in China are increasing year by year. Conquering cancer has become a difficult problem for human beings. Chemical drug therapy combined with molecular targeted therapy is a general and preferred anti-tumor clinical scheme, but the side effects and the drug resistance of cancer cells often hinder the efficacy. Therefore, it is of great significance to study the mechanism of drug resistance and the methods to reverse drug resistance. Chinese medicine has the characteristics of complex components, multiple targets, low toxicity, etc. A large number of experimental studies have demonstrated that the effective components or extracts of Chinese medicine can inhibit the proliferation, migration, and invasion of cancer cells, and induce apoptosis, autophagy, differentiation, and senescence. In clinical practice, Chinese medicine has been applied to the protection against ttumor, adjuvant treatment, and later consolidation. The research on Chinese medicine is expected to promote drug resistance reversal and cancer therapy. Studies have shown that the combination of Chinese medicine and chemotherapy can reverse drug resistance and increase efficacy, which has become the mainstream trend of cancer treatment. This study reviewed the mechanisms of the drug resistance of cancer cells induced by self-protective autophagy, gene mutation, high expression of enzymes, abnormal signaling pathways, and abnormal expression of RNA and protein, and summarized how compounds isolated from Chinese medicine, single drug and its extract, and classic anti-cancer prescription reversed the drug resistance to lay a solid foundation for the further investigation of the anti-tumor effect of Chinese medicine.

9.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 85-91, 2022.
Article in Chinese | WPRIM | ID: wpr-940292

ABSTRACT

ObjectiveTo investigate the effect of Draconis Sanguis petroleum ether fraction (DSPEF) on the proliferation, apoptosis, migration, and autophagy of human gastric cancer HGC-27 and MGC-803 cells, and preliminarily elucidate its molecular mechanism. MethodCell counting kit-8 (CCK-8) assay was used to detect the effect of DSPEF at different concentrations (0, 20, 40, 60, 80 mg·L-1) on the proliferation of HGC-27 and MGC-803 cells after 24, 48, 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment, respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells, respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h. ResultCompared with the control group, DSPEF(30 mg·L-1) inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner (P<0.05), and induced the apoptosis (P<0.01) and autophagy of HGC-27 and MGC-803 cells. DSPEF (60 mg·L-1) down-regulated the protein levels of phosphorylated mammalian target of rapamycin (p-mTOR) (P<0.05, P<0.01) and down-regulated phospho-signal transducer and activator of transcription 3 (p-STAT3) in HGC-27 and MGC-803 cells (P<0.01), suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway. ConclusionThe down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.

10.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 79-84, 2022.
Article in Chinese | WPRIM | ID: wpr-940291

ABSTRACT

ObjectiveTo investigate effect of aqueous extract of Trametes robiniophila (TRM,Huaier) on autophagy of human prostate cancer VCaP cells and Lamin B1 expression, so as to uncover its role in the proliferation of VCaP cells. MethodThe inhibitory effect of 0, 2, 4, 6, 8, 10 g·L-1 TRM aqueous extract on the proliferation of human prostate cancer VCaP cells at different time points were determined by cell counting kit-8 (CCK-8) assay. Acridine orange staining was conducted for analyzing the effect of TRM aqueous extract on the formation of autolysosomes in VCaP cells. After medication, the expression of microtubule-associated protein Ⅰ light chain 3 (LC3), autophagy-related protein 3 (Atg3), autophagy-related protein 5 (Atg5), and autophagy-related protein 7 (Atg7) in VCaP cells were detected by Western blot. The effect of TRM aqueous extract alone and its combination with autophagy inhibitor bafilomycin A1 on the proliferation of VCaP cells were assayed by CCK-8 assay. RNA interference technology was used to explore the role of Lamin B1 in anti-proliferation of VCaP cells by TRM. ResultCompared with the blank group, TRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells in a time- and concentration-dependent manner (P<0.01). Acridine orange staining showed that TRM aqueous extract promoted the formation of autolysosomes in VCaP cells. As revealed by Western blotting, TRM aqueous extract up-regulated the expression levels of LC3-Ⅱ, Atg3, Atg5, and Atg7 in contrast to those in the blank group (P<0.05). All these indicated that TRM aqueous extract induced the autophagy of VCaP cells. In addition, autophagy inhibition impaired the sensitivity of VCaP cells to TRM aqueous extract (P<0.05). The comparison with the blank group showed that TRM aqueous extract inhibited Lamin B1 protein expression in VCaP cells (P<0.01), which in turns weakened the sensitivity of VCaP cells to TRM aqueous extract. ConclusionTRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells possibly by inducing autography and down-regulating Lamin B1 expression. This study has provided a theoretical basis for the clinical application of TRM.

11.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 72-78, 2022.
Article in Chinese | WPRIM | ID: wpr-940290

ABSTRACT

ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila(Huaier) has proved to have anti-hepatoma activity, and this paper aims to explore the molecular mechanism. MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 (0, 0.05, 0.1, 0.25, 0.5, 1 g·L-1). Then cell survival was detected by methyl thiazolyl tetrazolium (MTT) and apoptosis by flow cytometry. In addition, expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1. ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group (P<0.01). After treatment with 1 g·L-1 TPG-1 for 48 h, the apoptosis rate of SK-HEP-1 cells increased (P<0.01), and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase (Caspase)-3 and Caspase-7, the key mediators of apoptosis (P<0.01). Additionally, TPG-1 (1 g·L-1) suppressed the migration of SK-HEP-1 cells (P<0.05). A total of 971 differentially expressed genes (DEGs) were identified in SK-HEP-1 cells after treatment with TPG-1, with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology (GO) terms of interleukin-6 (IL-6) biosynthesis, antigen processing and presentation, superoxide dismutase activity, positive regulation of mitogen-activated protein kinase kinase kinase (MAPKKK) cascade, nature killer (NK) cell chemotaxis, and chemokine biosynthesis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of nucleotide-binding oligomerization domain (NOD)-like receptor signaling pathway, apoptosis, Toll-like receptor signaling pathway, retinoic acid-inducible gene-Ⅰ (RIG-Ⅰ)-like receptor signaling pathway, T-cell receptor signaling pathway, and chemokine signaling pathway. Western blot results showed that TPG-1 (1 g·L-1) activated mitogen-activated protein kinase (MAPK) signaling pathway in SK-HEP-1 cells (P<0.01). ConclusionProteoglycan TPG-1 inhibited the proliferation and migration, and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.

12.
Journal of Biomedical Engineering ; (6): 84-91, 2022.
Article in Chinese | WPRIM | ID: wpr-928202

ABSTRACT

In order to improve the motion fluency and coordination of lower extremity exoskeleton robots and wearers, a pace recognition method of exoskeleton wearer is proposed base on inertial sensors. Firstly, the triaxial acceleration and triaxial angular velocity signals at the thigh and calf were collected by inertial sensors. Then the signal segment of 0.5 seconds before the current time was extracted by the time window method. And the Fourier transform coefficients in the frequency domain signal were used as eigenvalues. Then the support vector machine (SVM) and hidden Markov model (HMM) were combined as a classification model, which was trained and tested for pace recognition. Finally, the pace change rule and the human-machine interaction force were combined in this model and the current pace was predicted by the model. The experimental results showed that the pace intention of the lower extremity exoskeleton wearer could be effectively identified by the method proposed in this article. And the recognition rate of the seven pace patterns could reach 92.14%. It provides a new way for the smooth control of the exoskeleton.


Subject(s)
Humans , Algorithms , Exoskeleton Device , Lower Extremity , Motion , Support Vector Machine
13.
Chinese Journal of Biotechnology ; (12): 737-748, 2022.
Article in Chinese | WPRIM | ID: wpr-927740

ABSTRACT

CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.


Subject(s)
Artemisinins , Fermentation , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism
14.
Tissue Engineering and Regenerative Medicine ; (6): 887-893, 2021.
Article in English | WPRIM | ID: wpr-896406

ABSTRACT

BACKGROUND@#Bile duct injury (BDI), which may occur during cholecystectomy procedures and living-donor liver transplantation, leads to life-altering complications and significantly increased mortality and morbidity. Tissue engineering, as an emerging method, has shown great potential to treat BDI. Here, we aimed to explore the application of small intestinal submucosa (SIS) matrix composites with bone marrow mesenchymal stem cells (BMSCs) to treat BDI in a rabbit model. @*METHODS@#Rabbit-derived BMSCs were used as seed cells. Porcine SIS was used as the support material. Five centimetres of the common bile duct was dissected, and 1/3–1/2 of the anterior wall diameter was transversely incised to construct the rabbit BDI model. Then, SIS materials without/with BMSCs were inserted into the common bile duct of the BDI rabbits. After 1, 2, 4, and 8 weeks of implantation, the common bile duct was removed. Haematoxylin and eosin (HE) staining was used to assess pathological alterations in the common bile duct, while immunohistochemical staining and western blotting were used to detect expression of the epithelial cell markers CK19 and E-cadherin. Scanning electron microscopy was used to evaluate BMSC growth. @*RESULTS@#Compared with BMSCs alone, SIS-attached BMSCs had increased growth. HE staining showed that the injured bile duct healed well and that the complex gradually degraded as the time from implantation increased. Immunohistochemical staining and western blotting showed that compared with the control group, the in vivo complex group had significantly elevated expression levels of CK19 and E-cadherin. @*CONCLUSION@#BMSC implantation into SIS could improve BDI in rabbits, which might have clinical value for BDI treatment.

15.
China Journal of Orthopaedics and Traumatology ; (12): 321-327, 2021.
Article in Chinese | WPRIM | ID: wpr-879436

ABSTRACT

OBJECTIVE@#To explore the treatment strategy and clinical efficacy for os odontoideum complicated with atlantoaxial dislocation.@*METHODS@#The clinical data of 17 patients with os odontoideum complicated with atlantoaxial dislocation surgically treated from January 2006 to January 2015 were retrospectively analyzed, including 7 males and 10 females, aged 17 to 53 (43.1±11.3) years old;course of disease was 3 to 27(10.2±6.9) months. All patients received cranial traction before operation, 12 of 14 patients with reducible dislocation were treated by posterior atlantoaxial fixation and fusion, and 2 patients with atlantooccipital deformity were treated by posterior occipitocervical fixation and fusion;3 patients with irreducible alantoaxial dislocation were treated by transoral approach decompression combined with posterior atlantoaxial fixation and fusion. The operation time, intraoperative blood loss and perioperative complications were recorded. Visual analogue scale (VAS) and Japanese Orthopaedic Association (JOA) score were used to evaluate the change of neck pain and neurological function. Atlantoaxial joint fusion rate was evaluated by CT scan.@*RESULTS@#The operation time of posterior fixation and fusion ranged from 86 to 170 (92.2±27.5) min, and the intraoperative blood loss was 200-350 (250.7±65.2) ml. No vertebral artery injury and spinal cord injury were recorded. Among the patients underwent atlantoaxial fixation and fusion, 1 patient with reducible dislocation fixed by C@*CONCLUSION@#Surgical treatment of os odontoideum complicated with atlantoaxial dislocation can achieve satisfactory results, improve the patient's neurological function and improve the quality of life, however the surgical options needs to be individualized.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Atlanto-Axial Joint/surgery , Axis, Cervical Vertebra , Joint Dislocations/surgery , Quality of Life , Retrospective Studies , Spinal Fusion , Treatment Outcome
16.
Chinese Pharmacological Bulletin ; (12): 1524-1529, 2021.
Article in Chinese | WPRIM | ID: wpr-1014492

ABSTRACT

Aim To investigate the potential protective function of nervonic acid (NA) on the motor disorder in mice subjected to MPTP and the underlying mechanism. Methods The PD mice model was constructed and divided into control group, model group, nervonic acid (20 mg • k g

17.
Chinese Pharmacological Bulletin ; (12): 1213-1218, 2021.
Article in Chinese | WPRIM | ID: wpr-1014361

ABSTRACT

Aim To explore the effects of linoleic acid on the joint swelling in rats caused by rheumatoid arthritis (RA), and to reveal possible mechanism underlying the inhibitory effect of linoleic acid on TLR4/NF-κB signaling pathway in RA. Methods The RA rat model was constructed and divided into control group, model group, linoleic acid (0. 1 mL) group, linoleic acid (0.2 mL) group, linoleic acid (0.4 mL) group and methotrexate (MTX) group. After the model was successfully established, the corresponding drugs were given by gavage for seven days. The control group and the model group were given normal saline. The changes of rat joint swelling were measured; joint pathological damage was assessed by HE staining; the protein expression levels and the mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10 were determined by ELISA and qRT -PCR; the protein expression levels of TLR4 and p-p65 were determined by immunohisto-chemistry and Western blot. Results Compared with model group, linoleic acid significantly alleviated the joint swelling of RA rats; linoleic acid significantly inhibited the joint pathological damage, and markedly reduced the protein expression levels and mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10; linoleic acid inhibited the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway. Conclusions Linoleic acid inhibits the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway, thereby inhibiting the expression of inflammatory factors TNF-α, IL-1β, IL-6 and IL-10.

18.
Frontiers of Medicine ; (4): 438-447, 2021.
Article in English | WPRIM | ID: wpr-888752

ABSTRACT

Thoracic aortic dissection (TAD) without familial clustering or syndromic features is known as sporadic TAD (STAD). So far, the genetic basis of STAD remains unknown. Whole exome sequencing was performed in 223 STAD patients and 414 healthy controls from the Chinese Han population (N = 637). After population structure and genetic relationship and ancestry analyses, we used the optimal sequence kernel association test to identify the candidate genes or variants of STAD. We found that COL3A1 was significantly relevant to STAD (P = 7.35 × 10


Subject(s)
Humans , Aortic Dissection/genetics , Case-Control Studies , Cluster Analysis , Cohort Studies , Collagen Type III/genetics , Computational Biology , Genetic Predisposition to Disease
19.
China Journal of Chinese Materia Medica ; (24): 3330-3336, 2021.
Article in Chinese | WPRIM | ID: wpr-887982

ABSTRACT

The present study aimed to explore the correlation between agronomic traits and quality indexes of Dendrobium nobile and its application value in agricultural breeding. The cultivated strains of D. nobile in Hejiang-Chishui producing areas were extensively collected,and the main agronomic traits and quality indexes were measured. The agronomic traits with significant correlation with quality indexes were screened out by the correlation analysis,and then the parental lines and self-bred F_1 generation plants were furtherverified. Among 96 lines of D. nobile,the content of soluble polysaccharides showed a significant negative correlation with dendrobine( P < 0. 01),and no significant correlation with agronomic traits in stems and leaves. The content of dendrobine exhibited a significant positive correlation with the stem width-thickness ratio( at the largest cross section; P < 0. 01),and no significant correlation with other agronomic traits. Regression analysis further verified the positive correlation between dendrobine content and stem width-thickness ratio( R2> 0. 9). Two lines,JC-10 and JC-35,with significant differences in stem width-thickness ratio were screened out( P <0. 05). The corresponding F1 generation plants by self-pollination both showed that the dendrobine content was higher with greater stem width-thickness ratio( P < 0. 01). The experimental results suggested that within a certain range,the dendrobine content was higher in D. nobile with flatter stem. Therefore,in the breeding of D. nobile,this specific trait could be used for screening plants with high content of quality indexes such as dendrobine.


Subject(s)
Agriculture , Dendrobium/genetics , Plant Breeding , Plant Leaves/genetics , Polysaccharides
20.
China Journal of Chinese Materia Medica ; (24): 2037-2044, 2021.
Article in Chinese | WPRIM | ID: wpr-879127

ABSTRACT

As a traditional Chinese medicine, Chinese dragon's blood has multiple effects, such as activating blood to remove blood stasis, softening and dispelling stagnation, astringent and hemostasis, clearing swelling and relieving pain, regulating menstruation and rectifying the blood, so it is called "an effective medicine of promoting blood circulation". It has been widely used clinically to treat a variety of diseases. With the further research on Chinese dragon's blood, its anti-tumor medicinal value is gradually emerging. Modern pharmacological studies have shown that Chinese dragon's blood exerts anti-tumor effects mainly by inhibiting cell proliferation, inducing apoptosis, inducing DNA damage and cell cycle arrest, inducing senescence and autophagy of tumor cells, inhibiting metastasis and angiogenesis, as well as reversing multidrug resistance. This article focuses on the research progress on anti-tumor effects of Chinese dragon's blood extract and its chemical components, with a view to provide new references for the in-depth research and reasonable utilization of Chinese dragon's blood.


Subject(s)
Female , China , Dracaena , Plant Extracts , Resins, Plant
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