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1.
Acta Pharmaceutica Sinica ; (12): 507-513, 2022.
Article in Chinese | WPRIM | ID: wpr-922927

ABSTRACT

The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm, suitable cultivation area and the proper processing methods for herbal raw materials. Gentiana crassicaulis in Gentiana (Sect. Cruciata), Gentianaceae is one of the original plants of the Chinese herb Qinjiao (Gentianae Macrophyllae Radix), and its type specimen was collected in Lijiang, Yunnan. There is a long planting history of the herb in this area. In this study a sampling plot was designated in these traditional planting areas. G. crassicaulis was planted and herbal raw materials were harvested from the plot. The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as "sweating" or "no sweating", "slicing" or "no slicing" (whole root), and "stoving" or "no stoving" (air drying). The quality of all processed samples was evaluated. In addition, molecular markers were determined for identifying cultivated and wild samples from Lijiang, Yunnan. The results are as follows: ① Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis. ② A total of 270 sequences of trnC-GCA-petN, atpB-rbcL, psbN, ndhB-rps7 and ycf1 were obtained, and three genotypes were determined from the cultivated samples; the type III was shared by both cultivated and wild plants. Based on the molecular markers, a DNA barcoding method to identify cultivated and wild samples of G. crassicaulis from Lijiang, Yunnan was established. ③ Total content of loganic acid and gentiopicroside in all samples was ≥ 2.5%, and above the Chinese Pharmacopoeia (2020) limit. ④ In HPLC fingerprinting, 9 common peaks were assigned and similarity between all samples was > 0.999; and ⑤ In a PCA score plot all slice samples were clustered, while whole root samples were scattered. Therefore, our studies could provide basic data for optimizing the processing method, producing best quality Gentianae Macrophyllae Radix, and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.

2.
Article in Chinese | WPRIM | ID: wpr-888175

ABSTRACT

As the main chemical constituents, iridoids are widely distributed within Gentiana, Gentianaceae, with promising bioactivities. Based on the previous work, the transcriptome of G. lhassica, an original plant of Tibetan herb "Jieji Nabao", was sequenced and analyzed in this study, and the transcriptome databases of roots, stems, leaves, and flowers were constructed so as to explore unigenes that may encode the key enzymes in the biosynthetic pathway of iridoids. Then, qRT-PCR was used to validate the relative expression levels of 11 genes named AACT, DXS, MCS, HDS, IDI, GPPS, GES, G10H, 7-DLNGT, 7-DLGT, and SLS in roots, stems, leaves, and flowers. Also, the total contents of gentiopicroside and loganic acid were determined by HPLC, respectively. The results are as follows:(1)a total of 76 486 unigenes with an average length of 852 bp were obtained;(2)335 unigenes were involved in 19 stan-dard secondary metabolism pathways in KEGG database, with phenylpropanoid biosynthesis having the maximum number(75 unigenes), and no isoflavone biosynthetic pathway was annotated;(3)171 unigenes participatedin 27 key enzymes encoding in the biosynthetic pathway of iridoids, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR) gene was highly expressed;(4)qRT-PCR results were approximately consistent with RNA-Seq data and the relative expression levels of the 11 genes were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root);(5)the total contents of gentiopicroside and loganic acid were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root), and the difference was significant. This study provides basic scientific data for accurate species identification, evaluation of germplasm resources, research on secondary pro-duct accumulation of medicinal plants within Gentianaceae, and protection of endangered alpine species.


Subject(s)
Gene Expression Profiling , Gene Expression Regulation, Plant , Gentiana/genetics , Iridoids , Transcriptome
3.
Acta Pharmaceutica Sinica ; (12): 2005-2014, 2021.
Article in Chinese | WPRIM | ID: wpr-886999

ABSTRACT

italic>Gentiana crassicaulis Duthie ex Burk. in Gentiana (Sect. Cruciata), Gentianaceae, is one of the original plants of both Gentianae Macrophyllae Radix and Tibetan herb Jie-Ji Na-Bao, which contain such bioactive iridoids as gentiopicroside, loganic acid and others. In this study, based on previous work, the transcriptome of G. crassicaulis was sequenced and analyzed to construct transcriptome databases of roots, stems, leaves and flowers. qRT-PCR verification was conducted for parts of unigenes that may be key enzymes in the pathway of iridoid biosynthesis. The results are as follows: ① a total of 159 534 unigenes were obtained, with an average length of 679 bp. According to the functional classification of GO, unigenes can be divided into 3 categories with 67 branches. The unigenes were aligned in the KOG database and were classified into 25 categories according to function. ② In the KEGG database, 215 unigenes were implicated in 20 standard secondary metabolism pathways. The analysis shows that 305 unigenes encoded 28 key enzymes in the pathway of iridoid biosynthesis, and their expression in different organs is different; and ③ qRT-PCR was approximately consistent with RNA-Seq results. The 7 annotated unigenes identified in this study, HMGS, DXS, MCS, GPPS, G10H, 7-DLNGT and STR, all had higher relative expression levels in the above-ground parts (stem, leaf and flower) than in the underground part (root). Iridoids are common active and index components of such traditional Chinese medicines as Qinjiao, Longdan, Dangyao, and Qingyedan, among others. Therefore, this work provides basic scientific data for further development including obtaining active components or intermediates through biotechnology, exploring the accumulation of effective components, evaluating the quality of different ecotype varieties, and identifying authentic biosynthesis pathways of medicinal materials.

4.
Acta Pharmaceutica Sinica ; (12): 2584-2591, 2021.
Article in Chinese | WPRIM | ID: wpr-886949

ABSTRACT

As two original plants of Tibetan herb Jieji, Gentiana waltonii Burk. and Gentiana lhassica Burk. belong to Section Cruciata of Gentiana, Gentianaceae. Here, we report on whole chloroplast genome sequences in the alpine species, respectively, and the features of plastomes were investigated. The plastome of G. waltonii is 148 705 bp long (148 652 bp in G. lhassica) and encodes 112 genes, including 78 protein-coding genes, 30 transfer RNA genes, and 4 ribosomal RNA genes. Two pseudogenes, namely ψrps16 and ψinfA, were found in plastomes. In addition, two novel loci were detected, and a species-specific polymerase chain reaction assay was developed for differentiating G. waltonii and G. lhassica from 10 alpine species in Section Cruciata. Gentiana. Our study provides basic data for identifying Tibetan herbs, alpine species conservation and molecular phylogenetic studies of Gentiana and Gentianaceae.

5.
Article in Chinese | WPRIM | ID: wpr-871677

ABSTRACT

Objective:To analyze the risk factors for postoperative renal replacement therapy(RRT) in patients with acute Stanford type A aortic dissection. Develop and validate a prediction model based on the risk factors with the purpose of early intervention.Methods:A retrospective analysis of 215 patients who underwent surgery for acute Stanford type A aortic dissection in our hospital from April 2016 to April 2019 were performed. Clinical variables including age, gender, basal blood pressure, preoperative serum creatinine, intraoperative blood pressure, operation time, cardiopulmonary bypass time, aortic occlusion time, intraoperative blood transfusion(including autologous blood), intraocular fluid infusion, colloidal fluid infusion, intraoperative urine volume, bleeding volume, total fluid balance, and postoperative blood lactate value were collected and their association with renal replacement therapy were analysed. Clinical variables were screened using lasso regression. Applying the post-filtering variables to construct a predictive model, calculating the area under the receiver operating characteristic curve( AUC) of the predictive model and the sensitivity and specificity under the optimal threshold for model evaluation. Results:In the 215 patients with acute Stanford type A aortic dissection, 38 patients required renal replacement therapy, accounting for 17.67%. Preoperative serum creatinine, operation time, cardiopulmonary bypass time, aortic occlusion time, intraoperative blood pressure less than 80mmHg time, intraoperative blood pressure less than 55% of basal blood pressure time, intraoperative blood transfusion, intraoperative crystal fluid dosage, intraoperative urine volume and lactate value after ICU admission were important risk factors for postoperative renal replacement therapy(RRT) in patients with acute Stanford type A aortic dissection. The AUC for the predictive model established using these variables was 0.955(95% CI: 0.897-1.000). The specificity under the optimal threshold was 96.1% and the sensitivity was 90.9%. Conclusion:Perioperative clinical variables can predict the possibility of RRT in patients with acute Stanford type A aortic dissection after surgery, which may provide the possibility for early intervention.

6.
Journal of Chinese Physician ; (12): 549-554, 2020.
Article in Chinese | WPRIM | ID: wpr-867286

ABSTRACT

Objective:To detect serum thyroid hormone level, HBV-DNA expression level and liver function index in patients with chronic hepatitis B (CHB) in plateau Tibetan inhabited areas, and to analyze their correlations.Methods:From June 2016 to June 2018, 420 patients with HBsAg + CHB infection in our hospital were selected as the observation group, which was divided into Tibetan observation group ( n=300)and Han observation group ( n=120). According to the level of HBV-DNA, the observation group was divided into three groups: A, B and C group. The healthy people in the same period were selected as the control group, 220 cases in Tibetan control group and 120 cases in Han control group. The expression level of HBV-DNA was detected by fluorescence quantitative polymerase chain reaction (qRT-PCR). Serum levels of thyroid stimulating hormone (TSH), triiodothyronine (T3), tetraiodothyronine (T4), free triiodothyronine (FT3), and free tetraiodothyronine (FT4) were measured by chemiluminescence. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bile acid (TBA) levels were measured by automatic biochemical analyzer and kit; the prothrombin time (PT) was measured by hemagglutination analyzer. Results:The levels of serum ALT, AST, HBV-DNA, TBA, PT, laminin (LN), procollagen-Ⅲ (PC-Ⅲ), collagen-Ⅳ (C-Ⅳ) and hyaluronidase (HAC) in Tibetan and Han observation group were significantly higher than those in Tibetan and Han control group ( P<0.05). The level of serum HAC in Tibetan patients with CHB was significantly higher than that in Han CHB patients ( P<0.05); the level of serum ALT, AST, PT, TBA, C-Ⅳ, PC-Ⅲ, HAC, LN increased with the level of serum HBV-DNA in patients with CHB ( P<0.05); the level of serum TSH, T3, T4, FT3, FT4 and T3/T4 ratio in Tibetan and Han patients with CHB had no significant difference ( P>0.05). With the increase of serum HBV-DNA level in Tibetan patients, serum T3 level and T3/T4 ratio in group B and C were significantly lower than those in the control group ( P<0.05), T4 level was significantly higher than that in the control group ( P<0.05), and the levels of FT3, FT4 and TSH were not significantly different from those in the control group ( P>0.05). The results of Pearson test showed that serum T3 level and T3/T4 ratio were negatively correlated with ALT, AST, PT, TBA, LN, HBV-DNA, C-Ⅳ and HAC levels in Tibetan CHB patients ( P<0.05); serum T4 level was positively correlated with ALT, AST, PT, TBA, LN, HBV-DNA, C-Ⅳ and HAC levels in Tibetan CHB patients ( P<0.05). Conclusions:Thyroid hormones TSH, T3, and T4 levels in CHB patients in plateau Tibetan inhabited areas are decreased significantly with the increase of HBV-DNA levels, and they are negatively correlated with ALT, AST, PT and TBA levels, suggesting that HBV infection may cause liver injury to some extent, and then affect the level of thyroid hormone.

7.
Article in Chinese | WPRIM | ID: wpr-865624

ABSTRACT

Objective:To investigate the incidence of delirium and related risk factors in patients after cardiovascular surgery.Methods:From May 2012 to May 2019, 7 001 patients underwent cardiovascular surgery in the Nanjing First Hospital were retrospectively analyzed. The general clinical data, operation name, operation time, cardiopulmonary bypass time, aortic occlusion time, analgesic and sedative drugs use during postoperative ICU treatment, confusion assessment method for the intensive care unit (CAM-ICU) score, length of ICU stay, total hospital stay, adverse prognosis and postoperative delirium were recorded. The influence of postoperative delirium on hospital stay and adverse prognosis was analyzed. The risk factors of postoperative delirium were explored.Results:Among the 7 001 patients, 573 (8.18%) had postoperative delirium (delirium group), while 6 428 patients had no delirium (non-delirium group). The incidence of postoperative delirium in patients with acute aortic dissection (AAD) after Sun′s operation was significantly higher than that in patients with other cardiovascular surgery: 45.03% (204/453) vs. 5.64% (369/6 548), and there was statistical difference ( P<0.05). The age, length of ICU stay, total hospital stay, incidence of adverse prognosis, operation time, cardiopulmonary bypass time and aortic occlusion time in delirium group were significantly higher than those in non-delirium group: (59.72 ± 12.48) years vs. (58.81 ± 12.16) years, 5.49 (2.87, 9.49) d vs. 1.12 (0.90, 1.95) d, 21.92 (17.90, 28.22) d vs. 17.85 (14.93, 21.76) d, 7.33% (42/573) vs. 2.13% (137/6 428), (5.43 ± 2.51) h vs. (4.06 ± 1.33) h, (140.01 ± 55.13) min vs. (108.07 ± 42.98) min and (85.23 ± 37.30) min vs. (72.50 ± 34.15) min, and there were statistical differences ( P<0.01). Multivariant Logistic regression analysis result showed that intraoperative deep hypothermic circulatory arrest and selective cerebral perfusion was independent risk factor of postoperative delirium in patients with cardiovascular surgery ( OR = 10.922, 95% CI 7.444 to 16.120, P < 0.01). After excluding AAD patients, the incidences of postoperative delirium were 2.63% (11/418), 4.16% (34/817), 4.37% (71/1 625), 5.13% (122/2 379), 9.34% (114/1 221) and 19.32% (17/88) for patients<40 years, 40 to 49 years, 50 to 59 years, 60 to 69 years, 70 to 79 years and ≥ 80 years respectively. The incidence of postoperative delirium increased with age ( Z= 2.63, P= 0.009). The incidences of postoperative delirium were 1.47% (45/3 056), 3.22% (63/1 954), 5.69% (34/597), 12.14% (38/312), 18.18% (22/121), 22.62% (38/168), 25.93% (21/81) and 41.70% (108/259) for patients who stayed 1, 2, 3, 4, 5, 6, 7 and>7 d in ICU. The longer stay in ICU, the higher the incidence of postoperative delirium ( Z= 3.34, P = 0.001). Sequential organ failure score (SOFA) was used to evaluate the organ functions of patients. The scores of respiratory system, circulatory system, liver function and renal function in delirium group were significantly worse than those in non-delirium group, and there were statistical differences ( P<0.01); there was no significant difference in coagulation function between 2 groups ( P > 0.05). According to the use of analgesic and sedative drugs during the postoperative ICU stay, the patients were divided into dexmedetomidine alone group (3 355 cases) and dexmedetomidine combined with dezocine group (1 396 cases). The incidence of postoperative delirium in dexmedetomidine combined with dezocine group was significantly higher than that in dexmedetomidine alone group: 19.20% (268/1 396) vs. 5.66% (190/3 355), and there was statistical difference ( P<0.01). Conclusions:Age, operation time, extracorporeal circulation time, aortic occlusion time, intraoperative hypothermic circulatory arrest with selective cerebral perfusion, severity of disease and length of ICU stay are independent risk factors for postoperative delirium in patients after cardiovascular surgery. The choice of analgesic and sedative drugs during the perioperative period may affect the occurrence of postoperative delirium.

8.
Article in Chinese | WPRIM | ID: wpr-837555

ABSTRACT

@#Objective    To analyze whether hypernatremia within 48 hours after cardiac surgery will increase the incidence of delirium which developed 48 hours later after surgery (late-onset delirium). Methods    We conducted a retrospective analysis of 3 365 patients, including 1 918 males and 1 447 females, aged 18-94 ( 60.53±11.50) years, who were admitted to the Department of Cardiothoracic and Vascular Surgery of Nanjing First Hospital and underwent cardiac surgery from May 2016 to May 2019. Results    A total of 155 patients developed late-onset delirium, accounting for 4.61%. The incidence of late-onset delirium in patients with hypernatremia was 9.77%, the incidence of late onset delirium in patients without hypernatremia was 3.45%, and the difference was statistically different (P<0.001). The odds ratio (OR) of hypernatremia was 3.028 (95% confidence interval: 2.155-4.224, P<0.001). The OR adjusted for other risk factors including elderly patients, previous history of cerebrovascular disease, operation time, cardiopulmonary bypass time, lactate, hemoglobin≥100 g/L, prolonged mechanical ventilation, left ventricular systolic function, use of epinephrine, use of norepinephrine was 1.524 (95% confidence interval: 1.031-2.231, P=0.032). Conclusion    Hypernatremia within 48 hours after cardiac surgery may increase the risk of delirium in later stages.

9.
Acta Pharmaceutica Sinica ; (12): 1941-1950, 2020.
Article in Chinese | WPRIM | ID: wpr-825172

ABSTRACT

Jieji Nabao is a common Tibetan herb. According to our ethnobotanical studies, one of its original plants is identified as Gentiana crassicaulis Duthie ex Burk. (Gentianaceae). Endemic to the Qinghai-Tibet Plateau, this medicinal alpine plant is a threatened species. In this study, 163 individuals from 20 populations of G. crassicaulis were collected throughout its geographical range and amplified fragment length polymorphism (AFLP) was used to investigate genetic variation in this species. A cluster analysis was performed on the AFLP data with Halenia elliptica and Gentiana straminea as the outgroups. From 64 pairs of AFLP primer combinations, 12 pairs were selected for amplification and a total of 315 bands were amplified, of which 254 bands were polymorphic, accounting for 80.63%. High genetic differentiation was detected between populations (87%), and low within populations (13%). The UPGMA (unweighted pair-group method with arithmetic means) tree was topologically consistent with the traditional taxonomic treatments at the species level, and the populations of G. crassicaulis were divided into two branches: one from Yunnan and Guizhou, the other from Tibet, Qinghai, Sichuan and Gansu. PCA analysis and the Mantel test showed that there was a positive correlation between genetic distance and geographical distance. In addition, combined with SSR and SNP markers within cpDNA, the genetic differentiation within the Sichuan population S1 was validated.

10.
Chinese Medical Journal ; (24): 682-689, 2020.
Article in English | WPRIM | ID: wpr-878000

ABSTRACT

BACKGROUND@#Although a variety of risk factors of pneumonia after clipping or coiling of the aneurysm (post-operative pneumonia [POP]) in patients with aneurysmal subarachnoid hemorrhage (aSAH) have been studied, the predictive model of POP after aSAH has still not been well established. Thus, the aim of this study was to assess the feasibility of using admission neutrophil to lymphocyte ratio (NLR) to predict the occurrence of POP in aSAH patients.@*METHODS@#We evaluated 711 aSAH patients who were enrolled in a prospective observational study and collected admission blood cell counts data. We analyzed available demographics and baseline variables for these patients and analyzed the correlation of these factors with POP using Cox regression. After screening out the prognosis-related factors, the predictive value of these factors for POP was further assessed.@*RESULTS@#POP occurred in 219 patients (30.4%) in this cohort. Patients with POP had significantly higher NLR than those without (14.11 ± 8.90 vs. 8.80 ± 5.82, P 10 had significantly worse POP survival rate than patients having NLR ≤10. NLR at admission might be helpful as a predictor of POP in aSAH patients.


Subject(s)
Humans , Lymphocytes , Neutrophils , Pneumonia/etiology , Prognosis , Subarachnoid Hemorrhage , Treatment Outcome
11.
Article in English | WPRIM | ID: wpr-758951

ABSTRACT

Porcine epidemic diarrhea (PED) is a highly contagious enteric swine disease. The large economic impact of PED on the swine industry worldwide has made the development of an effective PED vaccine a necessity. S0, a truncated region of the porcine epidemic diarrhea virus (PEDV) spike protein, has been suggested as a candidate antigen for PED subunit vaccines; however, poor solubility problems when the protein is expressed in Escherichia coli, and the inherent problems of subunit vaccines, such as low immunogenicity, remain. Flagellin has been widely used as a fusion partner to enhance the immunogenicity and solubility of many difficult-to-express proteins; however, the conjugation effect of flagellin varies depending on the target antigen or the position of the fusion placement. Here, we conjugated flagellin, Vibrio vulnificus FlaB, to the N- and C-termini of S0 and evaluated the ability of the fusion to enhance the solubility and immunogenicity of S0. Flagellin conjugation in the presence of the trigger factor chaperone tig greatly improved the solubility of the fusion protein (up to 99%) regardless of its conjugation position. Of importance, flagellin conjugated to the N-terminus of S0 significantly enhanced S0-specific humoral immune responses compared to other recombinant antigens in Balb/c mice. The mechanism of this phenomenon was investigated through in vitro and in vivo studies. These findings provide important information for the development of a novel PED vaccine and flagellin-based immunotherapeutics.


Subject(s)
Animals , Diarrhea , Escherichia coli , Flagellin , Immunity, Humoral , In Vitro Techniques , Mice , Porcine epidemic diarrhea virus , Solubility , Swine , Swine Diseases , Vaccines, Subunit , Vibrio vulnificus , Vibrio
12.
Acta Pharmaceutica Sinica ; (12): 944-953, 2019.
Article in Chinese | WPRIM | ID: wpr-780207

ABSTRACT

The roots and flowers of Gentiana waltonii and Gentiana robusta are used as Tibetan herb Jie-Ji in traditional Tibetan medicine, with iridoids as the main active ingredient and index components. To study the pathway of iridoid biosynthesis, roots, stems, leaves and flowers of G. waltonii and G. robusta were subjected to a high-throughput transcriptomic sequencing analysis by Illumina HiseqXTen. After removing insignificant reads and de novo splicing, 79 455 and 78 466 unigenes were obtained from G. waltonii and G. robusta respectively, with average length as 834 bp and 862 bp. The unigene GO functions could be divided into three categories of 65 branches. The unigenes were aligned in KOG database and were classified into 25 classes according to function. In KEGG database, 315 and 340 unigenes of G. waltonii and G. robusta were implicated in 20 standard secondary metabolic pathways, respectively. Furthermore, 80 and 57 unigenes of the two species were analyzed to encode 24 key enzymes in the pathway related to iridoid biosynthesis. There were differences in gene expression among different organs. Based on sequence data, significant amounts of SSRs, SNPs and InDels were detected in each dataset. This study provides a platform for further development of molecular markers, excavation of functional genes, and research into metabolic pathways and their regulatory mechanism within G. waltonii and G. robusta.

13.
Acta Pharmaceutica Sinica ; (12): 166-172, 2019.
Article in Chinese | WPRIM | ID: wpr-778674

ABSTRACT

italic>Gentiana section Cruciata (Gentianaceae) is a medicinally important section of herbs, including Chinese traditional medicine Gentianae Macrophyllae Radix and Tibetan herb Jieji. Here, we assess the taxonomic significance using mtDNA nad1/b-c and nad5/d-e sequence data. A total of 144 nad1/b-c and nad5/d-e sequences from 11 species within Gentianaceae were obtained, including 138 sequences from 10 species within Gentiana section Cruciata and 6 sequences from Halenia elliptica (outgroup). The results showed that mtDNA nad1/b-c has species- level resolution within the section of Cruciata, i.e. the variable in the position 45 “C” could be used as a stable marker locus to distinguish G. robusta from other taxa; the variable in the position 352 and 353 “GA” could distinguish G. crassicaulis and G. tibetica from other taxa within the section. Intraspecies genotype variability was detected in nad1/b-c sequences of G. officinalis and G. siphonantha, respectively. These genotypes could be used as potential DNA barcode. In addition, intraspecies genotype variability was detected in nad5/d-e sequences of G. macrophylla, G. officinalis and G. siphonantha, respectively. Based on the stable marker locus, a species-specific PCR protocol was developed using the primer PF to identifying G. robusta in the section. This study could expand the understanding of the diversity of mtDNA nad1/b-c and nad5/d-e in the genus Gentiana, and provide the essence for the species identification within Gentiana section Cruciata.

14.
Article in Chinese | WPRIM | ID: wpr-744561

ABSTRACT

Objective To investigate the prediction by intravascular ultrasound (IVUS) with monocyte to high density lipoprotein-cholesterol (HDL-C) ratio (MHR) of 12-month prognosis in patients with intermediate non-left main coronary lesions after percutaneous coronary intervention (PCI). Methods Patients with intermediate non-left main coronary lesions diagnosed by coronary angiography were tested of monocyte counts and HDL-C levels at admission with MHRs calculated. IVUS was used to examine plaque stability in target lesions. Patients were dviided into stable plaque group (n=44) and unstable plaque group (n=140) according to the IVUS results. PCI was then operated in patients with unstable plaque or with minimum lumen area<4 mm2. The major adverse cardiovascular events (MACE) were recorded during the follow-up period of 12 months after PCI. Results MHR was significantly higher in unstable plaque group than that in stable plaque group[(22.6±8.4) vs.(14.1±7.2),P<0.001]. Receiver-operating characteristic (ROC) analysis revealed that an MHR cut-off of 16.05 had 74.2% sensitivity and 77.0% specificity for prediction of 12-month MACE after PCI (AUC 0.78, 95% CI 0.71–0.85, P<0.001). Besides, unstable plaque with MHR over 16.05 was an independent risk factor for 12-month MACE after PCI (adjusted HR 3.26, 95% CI 2.48–4.14, P=0.020). Conclusions IVUS combined with MHR is a valuable index predicting the prognosiso f patients with intermediate non-left main coronary lesions who underwent PCI.

15.
National Journal of Andrology ; (12): 322-328, 2019.
Article in Chinese | WPRIM | ID: wpr-816833

ABSTRACT

Objective@#To investigate the influence of the Rho/ROCK signaling pathway on the anti-cryodamage ability of human sperm and provide some theoretical evidence for the development of high-efficiency semen cryoprotectants.@*METHODS@#We collected semen samples from 25 healthy males, each divided into a fresh, a normal cryopreservation control and an Rho-inhibition group. Before and after freezing, we detected sperm motility, viability, membrane integrity, morphology, DNA fragmentation index (DFI), acrosomal enzyme activity (AEA) and mitochondrial membrane potential (MMP) and determined the expressions of RhoA and ROCK proteins in the sperm by immunofluorescence staining.@*RESULTS@#Compared with the normal cryopreservation control, the frozen-thawed sperm of the Rho-inhibition group showed significantly increased sperm motility ( [51.20 ± 7.70]% vs [57.50 ± 6.83]%, P = 0.002), survival rate ( [52.87 ± 5.07]% vs [60.24 ± 5.53]%, P = 0.001), membrane integrity ([59.78±5.56]% vs [67.10 ± 4.43]%, P = 0.001), percentage of morphologically normal sperm ([4.83 ± 1.11]% vs [7.46 ± 1.28], P = 0.001) and MMP (56.30 ± 4.28 vs 63.11 ± 2.97, P = 0.001), but decreased DFI ([27.64 ± 6.64]% vs [18.87 ± 4.07]%, P = 0.001). There was no statistically significant difference in the AEA of the frozen-thawed sperm between the control and Rho-inhibition groups (97.65 ± 9.31 vs 98.30 ± 11.33, P > 0.05). Immunofluorescence staining revealed extensive expressions of RhoA and ROCK proteins in the head and neck of the sperm.@*CONCLUSIONS@#The Rho/ROCK signaling pathway plays a role in the cryodamage to human sperm, and inhibiting the activity of Rho/ROCK can significantly improve the ability of sperm to resist cryodamage.

16.
Acta Pharmaceutica Sinica ; (12): 1009-1015, 2018.
Article in Chinese | WPRIM | ID: wpr-779964

ABSTRACT

As a common Tibetan herb, Bawo Sebo was mainly used in the treatment of rheumatoid arthritis and urarthritis in Traditional Tibetan medicine. Based on our ethnobotanical survey, the origin of the herb was determined as Swertia verticillifolia T. N. Ho et S. W. Liu (Gentianaceae), endemic to the region of the Qinghai-Tibet Plateau. The diagnostic characters:perennial; stem leaves in whorls; corolla campanulate, yellow-green, 4-lobed; nectary 1 per corolla lobe, naked. Also, its complete chloroplast (cp) genome was sequenced. It is 151 682 bp in length, including a large single copy (LSC) region of 82 623 bp, a small single copy (SSC) region of 18 335 bp and a pair of inverted repeats (IRs) of 25 362 bp. It contains 129 unique genes, including 84 protein-coding genes, 37 tRNAs and 8 rRNAs. This study provides information for understanding the diversity of Swertia cp genomes, and the alpine species identification, conservation and molecular phylogenetic researches of Swertia and Gentianaceae.

17.
Chinese Medical Journal ; (24): 330-338, 2018.
Article in English | WPRIM | ID: wpr-342043

ABSTRACT

<p><b>BACKGROUND</b>Mitofusin-2 (MFN2), a well-known mitochondrial fusion protein, has been shown to participate in innate immunity, but its role in mediating adaptive immunity remains poorly characterized. In this study, we explored the potential role of MFN2 in mediating the immune function of T lymphocytes.</p><p><b>METHODS</b>We manipulated MFN2 gene expression in Jurkat cells via lentiviral transduction of MFN2 small interfering RNA (siRNA) or full-length MFN2. After transduction, the immune response and its underlying mechanism were determined in Jurkat cells. One-way analysis of variance and Student's t-test were performed to determine the statistical significance between the groups.</p><p><b>RESULTS</b>Overexpression of MFN2 enhanced the immune response of T lymphocytes by upregulating Ca2+ (359.280 ± 10.130 vs. 266.940 ± 10.170, P = 0.000), calcineurin (0.513 ± 0.014 vs. 0.403 ± 0.020 nmol/L, P = 0.024), and nuclear factor of activated T cells (NFATs) activation (1.040 ± 0.086 vs. 0.700 ± 0.115, P = 0.005), whereas depletion of MFN2 impaired the immune function of T lymphocytes by downregulating Ca2+ (141.140 ± 14.670 vs. 267.060 ± 9.230, P = 0.000), calcineurin (0.054 ± 0.030 nmol/L vs. 0.404 ± 0.063 nmol/L, P = 0.000), and NFAT activation (0.500 ± 0.025 vs. 0.720 ± 0.061, P = 0.012). Furthermore, upregulated calcineurin partially reversed the negative effects of MFN2 siRNA on T cell-mediated immunity evidenced by elevations in T cell proliferation (1.120 ± 0.048 vs. 0.580 ± 0.078, P = 0.040), interleukin-2 (IL-2) production (473.300 ± 24.100 vs. 175.330 ± 12.900 pg/ml, P = 0.000), and the interferon-γ/IL-4 ratio (3.080 ± 0.156 vs. 0.953 ± 0.093, P = 0.000). Meanwhile, calcineurin activity inhibitor depleted the positive effects of overexpressed MFN2 on T cells function.</p><p><b>CONCLUSIONS</b>Our findings suggest that MFN2 may regulate T cell immune functions primarily through the Ca2+-calcineurin-NFAT pathway. MFN2 may represent a potential therapeutic target for T cell immune dysfunction-related diseases.</p>

18.
Article in Chinese | WPRIM | ID: wpr-657306

ABSTRACT

Objective:To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture. Methods:A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected. Results:Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05,P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05,P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (allP<0.05). Conclusion: EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.

19.
Chinese Journal of Zoonoses ; (12): 841-844, 2017.
Article in Chinese | WPRIM | ID: wpr-657269

ABSTRACT

Mycoplasma pneumonia is one of the important pathogens leading to respiratory tract infection.Recently,the incidence of M.pneumoniae infection increased rapidly,which contributes about 30% of lung disease in children.The early clinical symptoms for M.pneumoniae infection is not typical,which is not sensitive to antibiotics acting on cell walls.Therefore,early laboratory detection of M.pneumoniae is very important for later treatment.Herein,this paper aims to sum up the recent development progress of M.pneumoniae detection.

20.
National Journal of Andrology ; (12): 873-877, 2017.
Article in Chinese | WPRIM | ID: wpr-812865

ABSTRACT

Objective@#To investigate the role of pH2AX in the reversibility of mouse testicular reproductive function impaired by single heat stress.@*METHODS@#Twenty-four C57 male mice were randomly divided into heat stress and control groups and immersed in water at 43℃ and 25℃, respectively, for 15 minutes. At 1, 7, and 14 days of heat exposure, all the mice were sacrificed and their testis tissues collected for determining the apoptosis of the germ cells by TUNEL and measuring the expression level of the pH2AX protein by immunohistochemistry and Western blot.@*RESULTS@#The highest percentage of apoptotic cells were found in the seminiferous tubules of the mice in the heat stress group on the 1st day of the exposure and almost no apoptosis was observed at 7 and 14 days. The pH2AX protein was expressed in the nuclei of the basement membrane of adjacent seminiferous tubules. Compared with the control group, the expression of pH2AX was significantly increased on the 1st day of exposure (0.47 ± 0.02 vs 1.61 ± 0.04, P <0.01), then decreased at 7 days (0.85 ± 0.03) in comparison with that on the 1st day (P <0.01), and again elevated at 14 days (1.72 ± 0.02) as compared with either those at 1 and 7 days (P <0.01) or that of the control (P <0.01).@*CONCLUSIONS@#Heat stress causes dynamic changes of the pH2AX expression in the testis of the mouse, which are associated with heat stress-induced proliferation and division of the testicular spermatogenic cells.


Subject(s)
Animals , Apoptosis , Blotting, Western , Heat Stress Disorders , Histones , Metabolism , Hot Temperature , Immunohistochemistry , In Situ Nick-End Labeling , Male , Mice , Mice, Inbred C57BL , Random Allocation , Seminiferous Tubules , Cell Biology , Spermatozoa , Cell Biology , Metabolism , Testis , Time Factors
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