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1.
Article in Chinese | WPRIM | ID: wpr-1025086

ABSTRACT

Objective To analyze the effects of miR-181a-5p overexpression on metabolites in the small intestines of mice with subcutaneous oral cancer by detecting changes in metabolites and metabolic pathways.Methods Three groups were included in study:Control group,negative control and miR-181a-5p overexpression group.To establish a subcutaneous oral cancer model in mice,variously treated cell suspensions were subcutaneously injected into the upper right of the groin in female M-NSG severely immunodeficient mice.Changes in pathology and small intestinal tissues were assessed by HE staining.Changes in mouse body weight were also assessed.Tandem orbitrap mass spectrometry and ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry,were used to examine metabolites in the small intestines.By pre-analyzing the original data and quality rating sample data,XCMS was able to assess which metabolites were different among the groups.To identify unique metabolic pathways,KEGG enrichment analysis was used.Results A total of 170 distinct metabolites were found in the small intestinal tissues of Control and NC groups.Choline metabolism,alanine,aspartate,and glutamate metabolism,GABA synaptic metabolism,glycerophospholipid metabolism,cAMP signaling route,cancer center carbon metabolism,and niacin and niacin amine metabolic pathways were important signaling pathways for metabolite enrichment.In the NC group,16 distinct metabolites with VIP values larger than 2 were found in the small intestines compared with the OE group overexpressing miR-181a-5p.Glycerin phosphorylcholine,palmitic acid,3-hydroxybutyl carnitine,and β-hydroxybutyric acid were among the metabolites that significantly varied.The primary enhanced metabolic pathway was the choline pathway.Conclusions Mouse small intestines underwent slight changes from subcutaneous oral cancer with the greatest effect on metabolites critical for energy metabolism.The choline metabolic pathway was the pathway that selected absolutely metabolites in mouse small intestines with subcutaneous grafts of oral cancer.

2.
Article in Chinese | WPRIM | ID: wpr-1026880

ABSTRACT

Objective To observe the effects of electroacupuncture(EA)at"Hegu"and"Taichong"on the expressions of cytochrome C(Cyt-C)and Caspase-9 in the hippocampus of rats with chronic pain and depression comorbidity(CPDC);To explore its potential mechanism for the treatment of CPDC.Methods A total of 60 SD rats were randomly divided into sham-operation group,model group,medication group and EA group,with 15 rats in each group.The CPDC model was induced by twice injection of complete Freund's adjuvant into the plantar of the left hind paw.EA group was applied to bilateral"Hegu"and"Taichong"for 20 min,the medication group were treated with duloxetine suspension 10 mg/kg by gavage for 14 days.The mechanical paw withdrawal threshold and thermal withdrawal latency were measured on day 7,14,21 and 28 after the first injection,tail suspension test and sucrose preference test were performed on day 14 and 28,Nissl staining was used to observe hippocampal neurons and the number of Nissl body,the protein expressions of Cyt-C and Caspase-9 in hippocampal tissue were detected by Western blot and immunohistochemistry.Results Compared with the sham-operation group,the mechanical paw withdrawal threshold and thermal withdrawal latency of the model group significantly decreased(P<0.01),the tail suspension immobility time increased(P<0.01),the percentage of sucrose preference decreased(P<0.01),the neurons were thinly arranged,the neurons were damaged and lost,and the number of Nissl body were less(P<0.01),the apoptotic rate of hippocampal neurons increased(P<0.01),the expression of Cyt-C,Caspase-9 and cleaved-Caspase-9/Caspase-9 ratio in hippocampal tissue increased(P<0.01).Compared with the model group,the mechanical paw withdrawal threshold and thermal withdrawal latency of the EA group and medication group significantly increased(P<0.01),the tail suspension immobility time decreased(P<0.01),the percentage of sucrose preference increased(P<0.01),the loss of neurons in hippocampus was reduced,the cells were arranged neatly,and the nucleoli were clear,the number of Nissl body increased(P<0.01),the apoptotic rate of hippocampal neurons decreased(P<0.01),and the expression of Cyt-C,Caspase-9 and cleaved-Caspase-9/Caspase-9 ratio in hippocampal tissue decreased(P<0.01).Conclusion EA at"Hegu"and"Taichong"can alleviate pain and depression in CPDC rats,which may be related to inhibiting the expressions of Cyt-C and Caspase-9 in hippocampal tissue and inhibiting the apoptosis of hippocampal neurons,thus playing a neuroprotective effect.

3.
Article in Chinese | WPRIM | ID: wpr-1039119

ABSTRACT

General education in natural sciences is a critical component in higher education. In this article, I introduced the exploration and practice of curriculum construction of “Biological Clock and Health” at Tsinghua University. The course has been open to students for 3 semesters so far. Students who selected the course come from 27 departments of the university. The students provided positive feedbacks after taking this class, including self-report of improved sleep quality and better understanding of the research logic and methods in life sciences. The logic and methods of the construction of “Biological Clock and Health” described in this article provide insights to the construction of other courses in natural sciences of general education.

4.
Article in Chinese | WPRIM | ID: wpr-979226

ABSTRACT

ObjectiveTo compare the effects of two different insemination methods, conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), on pregnancy outcomes in patients with frozen-thawed D6(day 6) blastocyst transfer. MethodsA retrospective cohort study was conducted to analyze the clinical data of patients with thawed D6 blastocyst transfer between January 2018 and April 2020 at the Fertility Center of the Third Hospital of Sun Yat-sen University, divided into conventional IVF group (446 cycles ) and ICSI fertilization group (200 cycles) according to the fertilization method. Patients were divided into those with a history of D5(day 5) blastocyst transfer and those without. The patients’ general characteristics, blastocyst quality, and pregnancy outcomes of the two groups were compared. ResultsBMI, years of infertility, and basal FSH were not statistically significant in the IVF and ICSI groups (P > 0.05). Regardless of the history of D5 transfer, patients in the ICSI group were younger than those in the IVF group (P < 0.001), the proportion of primary infertility was significantly higher in the ICSI group (P < 0.001), and the number of oocytes obtained and the number of normally fertilized oocytes in the ICSI group were higher than those in the conventional IVF fertilization group (P < 0.001). The proportion of stage V and Ⅵ blastocysts was significantly higher in the conventional IVF group than in the ICSI group (21.6 % vs. 3.14 %, P < 0.001). High-quality blastocysts with an ICM score of A were significantly higher in the ICSI group than in the IVF group (23.8 % vs. 14.3 %, P = 0.01). The HCG-positive and clinical pregnancy rates were significantly higher in the ICSI group than in the IVF group (65.5 % vs. 48.4 %, P < 0.001; 56 % vs. 41.3 %, P = 0.001), and embryo implantation and live birth rates were also higher in the ICSI group than in the conventional IVF group (43.8 % vs. 30.9 %, P < 0.001; 43.0 % vs. 31.8 %, P = 0.006). After correcting for age and number of oocytes obtained between the two groups, the clinical pregnancy rate was still significantly higher in the ICSI group than in the conventional IVF group (OR: 1.590, 95 % CI: 1.030, 2.455, P = 0.036). Infant birth weight was lower in the ICSI group than in the IVF group (P = 0.016), and the differences in preterm birth rate, sex ratio, and mode of delivery were not statistically significant between the two groups. ConclusionsClinical pregnancy and live birth rates after thawing and transfer of D6 blastocysts fertilized by ICSI are higher than those of D6 blastocysts fertilized by conventional IVF, which may be related to the different factors contributing to the slow development of blastocysts in patients who received different fertilization methods. The relatively good pregnancy outcome after the transfer of thawed D6 blastocysts fertilized by ICSI may compensate to some extent for the difference in pregnancy outcome due to the relatively slow blastocyst development and a relatively higher proportion of D6 blastocysts after ICSI fertilization in male infertility patients.

5.
Article in Chinese | WPRIM | ID: wpr-1009165

ABSTRACT

OBJECTIVE@#To observe the clinical efficacy of intercondylar fossa plasty in preventing intercondylar fossa impingement syndrome after high tibial osteotomy.@*METHODS@#From August 2018 to August 2020, 84 patients with inverted knee osteoarthritis were treated by arthroscopy combined with high tibial osteotomy, and were divided into two groups with 42 cases in each group according to different surgical methods. In the intercondylar fossa plasty group, there were 13 males and 29 females, age ranged from 52 to 67 years old with an average of(58.27±4.32) years old, and arthroscopic intercondylar fossa plasty was performed first, and then high tibial osteotomy. In the arthroscopic cleansing group, 16 males and 26 females, age ranged from 50 to 71 years old with an average of (59.02±5.14) years old, underwent arthroscopic cleansing and then high tibial osteotomy. Postoperative treatment was evaluated using visual analogue scale(VAS), hospital for special surgery (HSS) score for the knee, and the occurrence of intercondylar percussa impingement.@*RESULTS@#All 84 patients were followed up, the duration ranged from 12 to 18 months with an average of (14.1±1.6) months. The VAS and HSS score of knee joint at 6, 12 and 18 months after surgery were significantly improved compared with preoperative period, and there was no significant difference between the two groups (P>0.05), but the incidence of intercondylar fossa index and intercondylar fossa impact between the two groups was significantly compared 18 months after surgery (P<0.05).@*CONCLUSION@#Intercondylar fossa plasty can effectively prevent the incidence of intercondylar fossa impact after high tibial osteotomy, and has a more significant effect on postoperative knee pain and function improvement.


Subject(s)
Male , Female , Humans , Middle Aged , Aged , Tibia/surgery , Osteoarthritis, Knee/surgery , Knee Joint/surgery , Treatment Outcome , Osteotomy/methods , Pain, Postoperative , Retrospective Studies
6.
Chinese Journal of Surgery ; (12): 791-797, 2023.
Article in Chinese | WPRIM | ID: wpr-985824

ABSTRACT

Objectives: To investigate the factors influencing the height of anterior peritoneal reflection (APR) for patients with rectal cancer, and to analyze the relationship between the APR and the lateral lymph node metastasis. Methods: Clinical data of 432 patients with tumor located within and below APR were retrospectively collected from the rectal cancer database at the Department of General Surgery, Peking Union Medical College Hospital from August 2020 to September 2022. Ninty-eight non-rectal cancer patients were also enrolled as a control group. There were 308 males and 124 females in the tumor group, aged (M(IQR)) 62 (16) years (range: 24 to 85 years) and 53 males and 45 females in the control group, aged 60 (22) years (range: 27 to 87 years). The APR height, pelvis, and tumor-related parameters were measured by MRI. A multifactor linear regression model was established to analyze the dependent correlation factors of APR height. These factors of the two groups were matched by propensity score matching and their APR heights were compared after matching. An ordinal Logistic regression model was established to explore the relationship between APR-related parameters and radiographic lateral lymph node metastasis. Results: The APR height of the tumor group was (98.7±14.4) mm (range: 43.3 to 154.0 mm) and the control group was (95.1±12.7) mm (range: 68.0 to 137.9 mm). Multivariable linear regression revealed that the greater the weight (B=0.519, 95%CI: 0.399 to 0.640, P<0.01), the anterior pelvic depth (B=0.109, 95%CI: 0.005 to 0.213, P=0.039) and the smaller the bi-ischial diameter (B=-0.172, 95%CI:-0.294 to -0.049, P=0.006), the higher the APR height. The tumor group had a higher APR height than the control group after propensity score matching ((98.3±14.2) mm vs. (95.1±12.7) mm, t=-1.992, P=0.047). Ordinal Logistic regression indicated that the longer segment of the tumor invade the nonperitoneal rectum was an independent influencing factor of radiographic lateral lymph node metastasis (OR=1.016, 95%CI: 1.002 to 1.030, P=0.021), while the distance between the anal verge and the tumor was not (OR=0.986, 95%CI: 0.972 to 1.000, P=0.058). Conclusions: The higher the weight, the deeper and narrower the pelvis, the higher the APR height. There is a certain relationship between APR and lateral lymph node metastasis on imaging.

7.
Article in Chinese | WPRIM | ID: wpr-992540

ABSTRACT

Objective:To investigate the risk factors of acute lung injury (ALI) caused by Staphylococcus aureus infection, and to construct a risk warning model. Methods:Patients with Staphylococcus aureus infection confirmed by sputum or blood culture admitted to the Affiliated Hospital of Jiangnan University from January 1, 2020 to May 10, 2022 were enrolled and divided into ALI group and non-ALI group. The age, smoking status, C-reactive protein (CRP), procalcitonin (PCT), neutrophil-to-lymphocyte ratio (NLR), albumin, oxygenation index and other clinical data were compared between the two groups. Univariate analysis was performed by using independent sample t test and chi-square test. Binary logistic regression analysis was used to screen the independent risk factors of ALI caused by Staphylococcus aureus infection, and a risk warning model was constructed. The receiver operator characteristic (ROC) curve was used to evaluate the predictive ability of the model. Results:There were 96 cases of Staphylococcus aureus infection, including 68 cases (70.8%) in ALI group, of which 41 cases (60.3%) were positive in sputum culture and 27 cases (39.7%) were positive in blood culture. Compared with the non-ALI group, the proportion of patients aged ≥60 years in ALI group was lower (58.8%(40/68) vs 64.3%(18/28)), the proportion of smoking was higher (58.8%(40/68) vs 35.7%(10/28)), and the differences were both statistically significant ( χ2=0.76 and 0.03, respectively, both P<0.05). The levels of CRP, PCT and NLR in the ALI group were all higher than those in non-ALI group, while oxygenation index and albumin level were both lower, and the differences were all statistically significant ( t=-5.28, -3.46, -9.87, 12.83 and 3.08, respectively, all P<0.05). Binary logistic regression analysis showed that CRP (odds ratio ( OR)=1.973, 95% confidence interval (95% CI) 0.956 to 2.989), PCT ( OR=3.734, 95% CI 1.014 to 13.746), NLR ( OR=1.152, 95% CI 1.058 to 2.254) and albumin ( OR=1.527, 95% CI 1.110 to 2.102) were independent risk factors for ALI caused by Staphylococcus aureus infection. The areas under the ROC curve of CRP, PCT, NLR, albumin and the risk warning model constructed from the combination of four risk factors were 0.69, 0.81, 0.83, 0.78 and 0.93, respectively. The sensitivities were 65.14%, 89.91%, 84.40%, 56.88% and 98.17%, respectively. The specificities were 62.37%, 60.22%, 65.59%, 80.64% and 93.55%, respectively. The accuracy of the effectiveness test of the risk warning model was 84.97%. Conclusions:CRP, PCT, NLR and albumin are the independent risk factors for ALI caused by Staphylococcus aureus infection. The risk warning model based on the above factors has a good early warning effect on ALI caused by Staphylococcus aureus infection.

8.
Article in Chinese | WPRIM | ID: wpr-993142

ABSTRACT

Objective:To investigate the clinical features, treatment options and prognostic factors of extramedullary plasmacytoma of the head and neck.Methods:Clinical data of 31 cases of extramedullary plasmacytoma of the head and neck treated in Eye Ear Nose and Throat Hospital of Fudan University from January 2005 to January 2020 were retrospectively analyzed. According to the involvement sites, the lesions were divided into the Waldeyer's ring involvement and non-involvement groups. The differences between two groups were compared by one-way ANOVA. The overall survival (OS) was analyzed by Kaplan-Meier method. The OS rate and local recurrence free survival (LRFS) rate between two groups were compared by log-rank test.Results:All 31 patients received radiotherapy. The median follow-up time was 98 months (7-192 months). Among them, 16 patients received radiotherapy and surgery, 10 patients received radiochemotherapy, and 2 patients received surgery and radiochemotherapy. Waldeyer's ring involvement was found in 15 patients, and Waldeyer's ring was not involved in 16 patients. The 10-year OS rate was 84.8%. Two cases showed local recurrence and 2 cases progressed to multiple myeloma. The 10-year OS rate in patients ≤55 years old was 100% vs. 67% in those >55 years old ( P=0.039). The 10-year OS rates in patients with and without Waldeyer's ring involvement were 93.3% vs. 75.2% ( P=0.031). The 10-year LRFS rate in patients with Waldeyer's ring involvement was 100% vs. 66.7% in those without Waldeyer's ring involvement ( P=0.022). The 10-year OS rate was 95% and 35.7% for patients with longest diameter of tumor ≤5 cm and >5 cm before radiotherapy, respectively ( P=0.02). Conclusions:Extramedullary plasmacytoma of the head and neck patients with longest diameter of tumor ≤5 cm obtain higher 10-year OS rate. Those with Waldeyer's ring involvement have better 10-year OS and LRFS rates.

9.
Chinese Pharmacological Bulletin ; (12): 1256-1262, 2023.
Article in Chinese | WPRIM | ID: wpr-1013753

ABSTRACT

Aim To explore the effect of Suanzaoren decoction(SZRD) on mitochondrial dysfunction in AD model of APP/PS1 mice via AMPK/SIRT1/PGC-1α signaling pathway and to reveal the possible mechanism. Methods Thirty APP/PS1 mice were randomly divided into app /PS1 group, low-dose SZRD group(L-SZRD) and high-dose SZRD group(H-SZRD). Ten C57BL/6JNju mice were set as control group(WT). Morris water maze test was used to detect the learning and memory ability of mice. Thioflavin T staining was used to observe senile plaques hippocampus. Immunohistochemistry was performed to detect the expression level of Aβ in hippocampus. Transmission electron microscope was used to observe the mitochondrial morph hology in hippocampus. Kits were employed to detect the contents of ATP and ROS in hippocampus; Western blot was employed to detect the expression levels of AMPK, p-AMPKThrK172, SIRT1, PGC-1α, NRF1, NRF2 and TFAM in hippocampus. Results Compared to the APP/PS1 group, L-SZRD and H-SZRD induced mouse cognitive impairment, reduced the deposition of senile plaques, inhibited the expression of Aβ, improved the damage of mitochondrial structure, increased the content of ATP in the hippocampus, reduced the expression level of ROS in hippocampus and increased the expression of p-AMPK-ThrK172, SIRT1, PGC-1α, NRF1, NRF2, TFAM Conclusions SZRD could improve the cognitive impairment, senile plaque deposition and mitochondrial dysfunction of AD mice, and its mechanism may be involved in the up-regulation of the expression of AMPK/SIRT1/PGC-1α signaling pathway.Reduced the Deposition of Senile Plaques, Inhibited the Expression of Aβ, Improved The Damage of Mitochondric Structure, Increased the Content of At in TH. E hippocampus, Reduced the Expression level of Ros in Hippocampus and Increased The Expression of P-Ampk-Thrk172, SIRT1, SIRT1 PGC-1α, NRF1, NRF2, TFAM. Conclusions SZRD could improve the cognitive impairment, senile plaque deposition and mitochondrial dysfunction of AD mice, and its mechanism may be involved in the up-regulation of the expression of AMPK/SIRT1/PGC-1α signaling pathway.Reduced the Deposition of Senile Plaques, Inhibited the Expression of Aβ, Improved The Damage of Mitochondric Structure, Increased the Content of At in TH. E hippocampus, Reduced the Expression level of Ros in Hippocampus and Increased The Expression of P-Ampk-Thrk172, SIRT1, SIRT1 PGC-1α, NRF1, NRF2, TFAM. Conclusions SZRD could improve the cognitive impairment, senile plaque deposition and mitochondrial dysfunction of AD mice, and its mechanism may be involved in the up-regulation of the expression of AMPK/SIRT1/PGC-1α signaling pathway.Senile plaque deposition and mitochondrial dysfunction of AD mice, and its mechanism may be involved in the up-regulation of the expression of AMPK/SIRT1/PGC-1α signaling pathway.Senile plaque deposition and mitochondrial dysfunction of AD mice, and its mechanism may be involved in the up-regulation of the expression of AMPK/SIRT1/PGC-1α signaling pathway.

10.
Chinese Pharmacological Bulletin ; (12): 672-678, 2023.
Article in Chinese | WPRIM | ID: wpr-1013808

ABSTRACT

Aim To investigate the effect of quercetin (Que) on secretory otitis media (SOM) rats and its mechanism. Methods Forty-eight male SD rats were selected and randomly divided into control group (control), model group (model), Que group (100 mg • kg

11.
Article in English | WPRIM | ID: wpr-1010988

ABSTRACT

Gut microbiota dysbiosis is an avenue for the promotion of atherosclerosis (AS) and this effect is mediated partly via the circulating microbial metabolites. More microbial metabolites related to AS vascular inflammation, and the mechanisms involved need to be clarified urgently. Paeonol (Pae) is an active compound isolated from Paeonia suffruticoas Andr. with anti-AS inflammation effect. However, considering the low oral bioavailability of Pae, it is worth exploring the mechanism by which Pae reduces the harmful metabolites of the gut microbiota to alleviate AS. In this study, ApoE-/- mice were fed a high-fat diet (HFD) to establish an AS model. AS mice were administrated with Pae (200 or 400 mg·kg-1) by oral gavage and fecal microbiota transplantation (FMT) was conducted. 16S rDNA sequencing was performed to investigate the composition of the gut microbiota, while metabolomics analysis was used to identify the metabolites in serum and cecal contents. The results indicated that Pae significantly improved AS by regulating gut microbiota composition and microbiota metabolic profile in AS mice. We also identified α-hydroxyisobutyric acid (HIBA) as a harmful microbial metabolite reduced by Pae. HIBA supplementation in drinking water promoted AS inflammation in AS mice. Furthermore, vascular endothelial cells (VECs) were cultured and stimulated by HIBA. We verified that HIBA stimulation increased intracellular ROS levels, thereby inducing VEC inflammation via the TXNIP/NLRP3 pathway. In sum, Pae reduces the production of the microbial metabolite HIBA, thus alleviating the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in AS. Our study innovatively confirms the mechanism by which Pae reduces the harmful metabolites of gut microbiota to alleviate AS and proposes HIBA as a potential biomarker for AS clinical judgment.


Subject(s)
Animals , Mice , Atherosclerosis/drug therapy , Diet, High-Fat , Endothelial Cells , Inflammation/drug therapy , Mice, Inbred C57BL , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Reactive Oxygen Species
12.
Acta Pharmaceutica Sinica B ; (6): 3535-3544, 2023.
Article in English | WPRIM | ID: wpr-1011113

ABSTRACT

C-Glycosides are important natural products with various bioactivities. In plant biosynthetic pathways, the C-glycosylation step is usually catalyzed by C-glycosyltransferases (CGTs), and most of them prefer to accept uridine 5'-diphosphate glucose (UDP-Glc) as sugar donor. No CGTs favoring UDP-rhamnose (UDP-Rha) as sugar donor has been reported, thus far. Herein, we report the first selective C-rhamnosyltransferase VtCGTc from the medicinal plant Viola tricolor. VtCGTc could efficiently catalyze C-rhamnosylation of 2-hydroxynaringenin 3-C-glucoside, and exhibited high selectivity towards UDP-Rha. Mechanisms for the sugar donor selectivity of VtCGTc were investigated by molecular dynamics (MD) simulations and molecular mechanics with generalized Born and surface area solvation (MM/GBSA) binding free energy calculations. Val144 played a vital role in recognizing UDP-Rha, and the V144T mutant could efficiently utilize UDP-Glc. This work provides a new and efficient approach to prepare flavonoid C-rhamnosides such as violanthin and iso-violanthin.

13.
Article in Chinese | WPRIM | ID: wpr-969893

ABSTRACT

The almost all guidelines of allergic rhinitis (AR) diagnosis and treatment in the world agree the strategy of "combination of prevention and treatment, four in one". There are more descriptions about anti-allergic medications and allergen immunotherapy (AIT), but less contents of environmental control and health education. It is necessary to emphasize again that clinicians must attach great importance to environmental control and strengthen health education in order to realize the three-level prevention of AR and reduce its harm.


Subject(s)
Humans , Rhinitis, Allergic/prevention & control , Desensitization, Immunologic/methods
14.
Article in Chinese | WPRIM | ID: wpr-982085

ABSTRACT

OBJECTIVE@#To screen better promoters and provide more powerful tools for basic research and gene therapy of hemophilia.@*METHODS@#Bioinformatics methods were used to analyze the promoters expressing housekeeping genes with high abundance, so as to select potential candidate promoters. The GFP reporter gene vector was constructed, and the packaging efficiency of the novel promoter was investigated with EF1 α promoter as control, and the transcription and activities of the reporter gene were investigated too. The activity of the candidate promoter was investigated by loading F9 gene.@*RESULTS@#The most potential RPS6 promoter was obtained by screening. There was no difference in lentiviral packaging between EF1 α-LV and RPS6-LV, and their virus titer were consistent. In 293T cells, the transduction efficiency and mean fluorescence intensity of RPS6pro-LV and EF1 αpro-LV were proportional to the lentiviral dose. The transfection efficiency of both promoters in different types of cells was in the following order: 293T>HEL>MSC; Compared with EF1 αpro-LV, RPS6pro-LV could obtain a higher fluorescence intensity in MSC cells, and RPS6pro-LV was more stable in long-term cultured HEL cells infected with two lentiviruses respectively. The results of RT-qPCR, Western blot and FIX activity (FIX∶C) detection of K562 cell culture supernatant showed that FIX expression in the EF1 α-F9 and RPS6-F9 groups was higher than that in the unloaded control group, and there was no significant difference in FIX expression between the EF1 α-F9 and RPS6-F9 groups.@*CONCLUSION@#After screening and optimization, a promoter was obtained, which can be widely used for exogenous gene expression. The high stability and viability of the promoter were confirmed by long-term culture and active gene expression, which providing a powerful tool for basic research and clinical gene therapy of hemophilia.


Subject(s)
Humans , Transduction, Genetic , Genetic Vectors , Hemophilia A/genetics , Transfection , Blood Coagulation Factors/genetics , Lentivirus/genetics
15.
Article in Chinese | WPRIM | ID: wpr-1045878

ABSTRACT

Objective: To investigate the species, concentration and seasonal trends of main airborne allergenic pollen in 4 districts and 5 counties of Hohhot City. Methods: The Department of allergy, Beijing Shijitan Hospital Affiliated to Capital Medical University conducted a cross-sectional study about monitoring the airborne allergenic pollen from August 1, 2021 to July 31, 2022 by the gravitational method in 4 districts and 5 counties of Hohhot City, which include Yuquan District, Xincheng District, Huimin District, Saihan District, Tuoketuo County, Helingeer County, Tumotezuoqi County, Wuchuan County and Qingshuihe County. Daily pollens were counted and identified by optical microscopy, and the data were analyzed. Results: The airborne allergenic pollen was collected every month all year round in 4 districts and 5 counties of Hohhot city. Through the whole year of the total quantity of pollens ranged from 24 850 to 50 154 grains per 1 000 mm2 and two peaks of pollen concentration in air were observed,which happened in spring (from March to May) and in summer and autumn (from July to September). In spring, the main pollens were tree pollens, which principally distributed in Populus pollen (18.29%), Ulmus pollen (8.36%), Pinus pollen (6.20%), Cupressaceae pollen (5.23%), Betulaceae pollen (2.73%), Salix pollen (1.80%) and Quercus pollen (1.16%). In summer and autumn, the main pollens were weed pollens, which mainly included Artemisia pollen (42.73%), Chenopodiaceae pollen or Amaranthaceae pollen (7.46%), Poaceae pollen (2.26%), Humulus pollen or Cannabis pollen (0.60%). Conclusion: There were two peaks of main airborne allergenic pollen in 4 districts and 5 counties of Hohhot City. In the spring peak of pollen, the main airborne pollens were tree pollens. In the summer and autumn peak of pollen, the main airborne pollens were weed pollens. The Artemisia pollen was the most major airborne pollen in this area.


Subject(s)
Humans , Cross-Sectional Studies , Pollen , Hospitals
16.
Article in Chinese | WPRIM | ID: wpr-1046201

ABSTRACT

Objective: To investigate the species, concentration and seasonal trends of main airborne allergenic pollen in 4 districts and 5 counties of Hohhot City. Methods: The Department of allergy, Beijing Shijitan Hospital Affiliated to Capital Medical University conducted a cross-sectional study about monitoring the airborne allergenic pollen from August 1, 2021 to July 31, 2022 by the gravitational method in 4 districts and 5 counties of Hohhot City, which include Yuquan District, Xincheng District, Huimin District, Saihan District, Tuoketuo County, Helingeer County, Tumotezuoqi County, Wuchuan County and Qingshuihe County. Daily pollens were counted and identified by optical microscopy, and the data were analyzed. Results: The airborne allergenic pollen was collected every month all year round in 4 districts and 5 counties of Hohhot city. Through the whole year of the total quantity of pollens ranged from 24 850 to 50 154 grains per 1 000 mm2 and two peaks of pollen concentration in air were observed,which happened in spring (from March to May) and in summer and autumn (from July to September). In spring, the main pollens were tree pollens, which principally distributed in Populus pollen (18.29%), Ulmus pollen (8.36%), Pinus pollen (6.20%), Cupressaceae pollen (5.23%), Betulaceae pollen (2.73%), Salix pollen (1.80%) and Quercus pollen (1.16%). In summer and autumn, the main pollens were weed pollens, which mainly included Artemisia pollen (42.73%), Chenopodiaceae pollen or Amaranthaceae pollen (7.46%), Poaceae pollen (2.26%), Humulus pollen or Cannabis pollen (0.60%). Conclusion: There were two peaks of main airborne allergenic pollen in 4 districts and 5 counties of Hohhot City. In the spring peak of pollen, the main airborne pollens were tree pollens. In the summer and autumn peak of pollen, the main airborne pollens were weed pollens. The Artemisia pollen was the most major airborne pollen in this area.


Subject(s)
Humans , Cross-Sectional Studies , Pollen , Hospitals
17.
Article in Chinese | WPRIM | ID: wpr-1025009

ABSTRACT

Objective To observe the effects of plantar injection of complete Freund's adjuvant(CFA)on pain-depressive behavior and changes in hippocampal monoamine neurotransmitters in rats to establish an animal model of related comorbidity.Methods Sixteen male,8-week-old,SPF-grade healthy SD rats were randomly divided into model and control groups with eight rats in each group.In the model group,rats were anesthetized and injected with 100 μL CFA in the left hind paw to induce the comorbid chronic inflammatory pain and depression model.In the control group,rats were injected with the same volume of saline.Pain thresholds were measured using the Von Frey hair and thermal radiation instrument,and depression-like behaviors were assessed using the open field test(OFT),tail suspension test(TST),and forced swim test(FST).Enzyme-linked immunosorbent assays were used to measure 5-hydroxytryptamine(5-HT),dopamine(DA),and norepinephrine(NE)in rat hippocampal tissue.Histological changes in the hippocampal area were observed by hematoxylin-eosin(HE)staining.Results Compared with the control group,the mechanical withdrawal threshold and thermal withdrawal latency in the model group were significantly decreased at 3,7,and 14 days(P<0.01).The total distance in the OFT was significantly reduced at 7 and 14 days(P<0.01),and the time spent in the center zone was significantly decreased at 14 days(P<0.01).Immobility time in the TST was significantly increased at 14 days(P<0.01),and the immobility time in the FST was significantly increased at 7 and 14 days(P<0.05,P<0.01).5-HT,DA,and NE contents in hippocampal tissue of the model group rats were significantly reduced compared with those in the control group(P<0.01),and hippocampal tissue in the model group showed pathological changes,including irregular neuronal shapes,loose and disordered arrangement,increased intercellular space,some unclear cell nuclei,and some neuronal contraction and apoptosis.Conclusions Injection of 100 μL CFA into the footpad causes pain hypersensitivity,depression-like behavior,significant reductions in monoaminergic neurotransmitters in the hippocampus,and histological changes in the hippocampus,effectively simulating the manifestations of comorbid pain and depression,and is an experimental model to study the pathological mechanisms of comorbid chronic inflammatory pain and depression.

18.
Article in Chinese | WPRIM | ID: wpr-954205

ABSTRACT

Objective:To study the differentially expressed genes in chronic periodontitis (CP) and to explore the correlation with disease severity.Methods:Gene expression profile data associated with CP were screened in the Gene Expression Omnibus (GEO) database and analyzed with GEO2R online software to create volcano maps. Kyoto Encyclopedia of Genes and Genomes (KEEG) and Gene Ontology (GO) analyses were performed on the screened CP-associated differentially expressed genes to predict their possible functions and signaling pathways. The protein-protein interaction database (STRING) was used to analyze the interaction relationships between the encoded proteins of the screened CP-related differentially expressed genes. Cytohubba software was used to identify key genes in the signaling pathway. One120 CP patients and 40 healthy controls were selected. The screened CP-related genes were validated by the real-time polymerase chain reaction (q-PCR) method.Results:A total of 1 151 CP differentially expressed genes that met the requirements were screened. These genes were mainly enriched in the GO pathway for positive regulation of granulocyte differentiation, helper T-cell differentiation, leukocyte aggregation, regulation of acute inflammatory response, chemokine-mediated and endoplasmic reticulum unfolded protein response, as well as in the KEGG pathway for NFB pathway, chemokine pathway, cytokine receptor interaction, leukocyte transendothelial migration pathway, B-cell receptor pathway, Toll-like receptor pathway, etc. The protein-protein interaction network was constructed using the screened CP-related differentially expressed genes, which included 78 nodes and 496 links, with a mean aggregation coefficient and mean connectivity of 0.69 and 12.7, respectively. Cytohubba analysis showed that Sell was a key gene in the signaling pathway, and its relative expression levels in the gingival fluids of the three CP groups with different degrees(1.14±0.46, 0.86±0.41, 0.52±0.46) was significantly lower than that of the control group (1.50±0.65) (all P<0.05). The area under the ROC curve (AUC) of subjects diagnosed with CP using Sell expression levels in gingival fluid was 0.79 (95% CI: 0.71 to 0.86). The AUC values were greater than 0.65 at 95% CI when Sell was used as a biological marker to evaluate the severity of CP. Conclusions:CP-related differentially expressed genes are mainly enriched in the number of pathways associated with the inflammatory response of periodontitis. The expression levels of Sell genes were significantly reduced in the gingival sulcus fluid of CP patients and correlated with the severity of the disease. The Sell genes are expected to be a biomarker for CP grading.

19.
Article in Chinese | WPRIM | ID: wpr-936237

ABSTRACT

Objective: To investigate the sensitization characteristics of Juniperus chinensis pollen in patients with allergic rhinitis and/or allergic asthma in Beijing area, and to explore the characteristics of Juniper chinensis pollen sensitized population. Methods: Patients with suspected allergic rhinitis and/or asthma from January 2017 to December 2019 in the outpatient department of Allergy Department of Beijing Shijitan Hospital were selected in this study. Skin prick test (SPT) was performed with Juniper chinensis pollen allergen reagent to compare different age and disease allergen distribution, and to observe the sensitization characteristics of its population. All of the analyses were performed using SAS software version 9.4. Results: A total of 8 380 patients were enrolled in the end. The total positive rate of Juniper chinensis pollen SPT reached 49.92% (4 183/8 380). The positive rate of Juniper chinensis pollen SPT was highest in the 10-14 age group, reaching 60.99% (283/464). Compared with other age groups, there was a statistical difference (χ²=266.77, P<0.01). The SPT positive rate of patients aged less than 10 years increased with the increase of age, while the SPT positive rate of patients aged over 40 years decreased with the increase of age. Single Juniper chinensis pollen was less allergenic, accounting for about 25.05% (1 048/4 183), and the patients' age was (35.21±12.39) years. Regardless of single Juniper chinensis pollen or other pollen allergies, allergic rhinitis was the main disease. Among the patients with SPT positive Juniper chinensis pollen combined with other inhaled pollen allergens, willow pollen accounted for the first (74.99%). The positive rate of Juniper chinensis pollen was the highest in patients with single allergic rhinitis, accounting for 52.05% (3 797/7 295), and the rate in patients with single allergic asthma was the lowest, accounting for 17.49% (53/303), with statistically difference (χ²=138.99, P<0.01). Conclusions: Juniper chinensis pollen is highly sensitized in patients with allergic rhinitis and/or allergic asthma in Beijing . The positive rate of SPT is highest among 10-14 age group, most of which showed strong positive reaction, and allergic rhinitis is more common in Juniper chinensis pollen sensitization diseases.


Subject(s)
Adolescent , Adult , Child , Humans , Allergens , Asthma , Juniperus , Pollen , Rhinitis, Allergic , Skin Tests
20.
Acta Pharmaceutica Sinica ; (12): 2334-2341, 2022.
Article in Chinese | WPRIM | ID: wpr-937038

ABSTRACT

Huachansu is a traditional Chinese medicine widely used in the clinic for cancer therapy, while the underlying mechanism is not fully clarified. This study was to investigate the targets and mechanisms of cinobufagin (CBG), an active component of Huachansu, in terms of blocking mitosis of cancer cells. Propidium iodide (PI) DNA staining was used to analyze the effect of CBG on cell cycle. The effect of CBG on mitosis of cancer cells was examined by α-tubulin and pericentrin staining after synchronization by a double thymidine block. Tubulin turbidity, tubulin polymerization and α‍-tubulin immunofluorescence assays were used to evaluate the effect of CBG on microtubule polymerization. CRISPR/Cas9 gene-editing technology was used to knockout microtubule-severing protein Katanin regulatory subunit B1 (KATNB1) in HCT116 cells, and the inhibitory effect of CBG on wild-type cells and knockout cells was measured by CCK-8. The engagement of CBG with KATNB1 was measured by CETSA and DARTS assays. The effect of CBG on KATNB1 protein and mRNA level was examined by Western blot and real-time PCR, respectively. Our data showed that CBG arrested HCT116 cell cycle at the G2/M phase, disrupted mitosis and induced centriole overduplication. CBG significantly inhibited tubulin polymerization in vitro and in vivo. The cytotoxicity of CBG inhibition on HCT116 was significantly attenuated upon KATNB1 depletion. Moreover, CBG bound to KATNB1 and decreased its protein level, while mutated KATNB1 weakened this effect. In conclusion, CBG inhibited microtubule polymerization via targeting KATNB1, thereby disrupting mitosis in cancer cells.

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