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1.
Acta Pharmaceutica Sinica ; (12): 482-488, 2024.
Article in Chinese | WPRIM | ID: wpr-1016642

ABSTRACT

The gene GeDRP1E encoding dynamin-related protein 1E in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDRP1E gene was carried out by using ExPASy, ClustalW, MEGA, etc. Positive transgenic Arabidopsis plant and potato minituber were obtained with the genetic transformation system of Arabidopsis and potato. The plant height and seed setting rate of transgenic Arabidopsis, and agronomic characters, such as size, weight and starch content of potato minituber of transgenic potato were tested and analyzed. And GeDRP1E gene function was preliminarily investigated. The results showed that the open reading frame of GeDRP1E gene was 1 899 bp in length and 632 amino acids residues were encoded, with a relative molecular weight of 69.90 kDa and a molecular formula of C3079H4973N883O933S19. It was predicted that the theoretical isoelectric point was 7.27, the instability coefficient was 43.34, and the average hydrophilicity index was -0.259, which was indicative of an unstable hydrophilic protein. GeDRP1E has no transmembrane structure and signal peptide, and was localized in the cytoplasm. The phylogenetic tree showed that GeDRP1E was highly homologous with DRP1E proteins of other plant species, among which GeDRP1E had the highest homology with DcDRP1E (XP_020689662.1) in Dendrobium candidum, reaching 90.05%. GeDRP1E plant expression vector pCambia1300-35Spro-GeDRP1E was constructed by double digests, and Arabidopsis complementary mutant and potato overexpression strain of GeDRP1E gene were obtained by Agrobacterium-mediated gene transformation. Compared with the Arabidopsis AtDRP1E mutant, the height and seed setting rate of the GeDRP1E complementation mutant were rescued. The minituber of GeDRP1E overexpression potato had larger size, heavier weight and higher starch content, comparing to wild-type potato. It was preliminarily induced that GeDRP1E was involved in mitochondrial morphology regulation, which related to the growth and development of Arabidopsis plants and potato miniature. The research results laid a foundation for further elucidating the molecular mechanisms underlying the growth and development of G. elata tuber development.

2.
China Pharmacy ; (12): 890-895, 2024.
Article in Chinese | WPRIM | ID: wpr-1013556

ABSTRACT

Tyrosine kinase inhibitors (TKIs) represent a class of small-molecule targeted drugs that improve the survival time of patients with gastrointestinal stromal tumor (GIST). Imatinib, sunitinib, regorafenib, ripretinib, and avapritinib are commonly used TKIs in the clinical treatment of various types of GIST. This article provides a comprehensive review of the pharmacokinetics and therapeutic drug monitoring (TDM) of these five drugs, finding that there is significant individual variability in the pharmacokinetics of these drugs. Among them, the absorption of imatinib, regorafenib, and avapritinib are influenced by food intake. Imatinib should be taken with meals and 200 mL of water, regorafenib is taken with a low-fat meal, while avapritinib is taken on an empty stomach. TKIs are mainly metabolized by cytochrome P450 3A4 (CYP3A4), and when used in combination with CYP3A4 inducers or inhibitors, drug exposure levels will significantly change; apart from metabolic enzymes, the exposure levels of TKIs are also influenced by interactions with the transporter proteins P-glycoprotein and breast cancer resistance protein. Currently, research on TDM for TKIs is still in the exploratory stage, with a substantial amount of literature reporting the effective concentrations of imatinib, sunitinib and regorafenib. However, the precise relationship between exposure levels and efficacy/ toxicity needs further exploration. Currently, there is a lack of research on the correlation between exposure levels and efficacy/ toxicity of ripretinib and avapritinib. It is recommended to implement TDM in patients taking these drugs and explore their therapeutic window in combination with pharmacokinetic models. The commonly used methods for clinical TDM of TKIs include immunoassay, chromatography, and surface-enhanced Raman spectroscopy, providing a technical basis for clarifying the therapeutic window of TKIs.

3.
JOURNAL OF RARE DISEASES ; (4): 57-62, 2024.
Article in English | WPRIM | ID: wpr-1006918

ABSTRACT

ObjectiveTuberous sclerosis complex (TSC) is a rare autosomal dominant genetic disease involving multiple organ system. Renal angiomyolipoma (RAML) is a leading cause of death for adult TSC patients. Our study aims at investigating the clinical manifestations of TSC-RAML to enable clinicians to have a better understanding of the disease in Chinese patients.MethodsWe reviewed retrospectively the data collected from patients with TSC-RAML in Peking Union Medical College Hospital from January 2014 to January 2023. We also collected information about the age, gender, imageological examination, RAML stage, and comorbidities.ResultsA total of 186 TSC-RAML patients were registered, 65 of whom were males and 121 were females(male-to-female ratio 1∶1.86). The median age of all patients was 31 years old. Totally, 117 cases (62.9%)of RAML were rated stage 6. Twenty-two cases (11.8%) had a history of tumor rupture and bleeding. Research shows statistical differences between high and low grade on RAML rupture bleeding(P=0.0475). Angiofibromas/fibrous cephalic plaque (155/186, 83.3%), subependymal nodules(103/146, 70.5%), lymphangioleiomyomatosis (102/157, 65.0%), hypomelanotic macules (114/186, 61.3%), and shagreen patch (83/186, 44.6%)were the most common clinical manifestations. All patients with lymphangioleiomyomatosis(LAM) were female(P < 0.0001).ConclusionsFemale patients predominated TSC-RAML patients. Most RAML were in stage 6. About 11.8% cases had a history of tumor rupture and hemorrhage. The higher grade meant the higher proportion of tumor rupture and hemorrhage. Dermatological and nervous lesions were the most common comorbidities. All patients with LAM in this study were female.

4.
Chinese Journal of Clinical Pharmacology and Therapeutics ; (12): 283-289, 2023.
Article in Chinese | WPRIM | ID: wpr-1014669

ABSTRACT

AIM: Previous studies have suggested that big stick design (BSD) method can only be used in clinical trials of two treatments with equal proportion, which has good statistical performance and has become the recommended choice of randomized methods. This study expands BSD method, so that it can be applied to three groups, and provides more randomized methods for clinical trials. METHODS: On the basis of BSD method used in two treatments with equal proportion, the derivation conditional allocation probability of BSD method used in three treatments with equal proportion was carried out. BSD method was compared with simple randomization (SR) method, permuted block design (PBD) method and block urn design (BUD) method by Monte-Carlo simulation in balance and randomness. RESULTS: In terms of balance, PBD method was the best, followed by BUD method, BSD method, and SR method was the worst. In terms of randomness, SR method was the best, followed by BSD method, BUD method and PBD method. The comprehensive performance showed that BSD method was better than BUD method, PBD method and SR method. CONCLUSION: The expanded BSD method used in three treatments with equal proportion has good comprehensive performance, and it can be the recommended randomization method for clinical trials of three treatments with equal proportion.

5.
Chinese Pharmacological Bulletin ; (12): 212-218, 2023.
Article in Chinese | WPRIM | ID: wpr-1013845

ABSTRACT

Malignant tumor is one of the important reasons threatening human health and safety at present. The application of antiangiogenic drugs and immune checkpoint inhibitors has brought great hope for tumor treatment, but the complex interlaced relationship between tumor blood vessels and immune microenvironment leads to unsatisfactory efficacy. In addition,VEGF, a key driver of tumor angiogenesis, interferes with the maturation of dendritic cells, thereby inhibiting the initiation of T cells. VEGF also induces depletion of CD8+T cells. At the same time, various innate and acquired immune cells secrete angiogenic factors that accelerate uncontrolled angiogenesis and promote vascular immaturity. Therefore targeting tumor blood vessels and immunity is a potential strategy for enhancing tumor immunotherapy. In recent years numerous studies have found that using the blood vessels and the immune intervention strategies combined with antitumor immune therapy has achieved good results. In this review the immune and vascularized tumor microenvironment are reviewed and discussed, and the research progress of vascularized immune interintervention strategy for tumor treatment in recent years is reviewed so as to provide reference for further improving the efficacy of tumor immunotherapy.

6.
Chinese Pharmacological Bulletin ; (12): 463-469, 2023.
Article in Chinese | WPRIM | ID: wpr-1013831

ABSTRACT

Aim To explore the effect of γ-ray on the mRNA,protein expression levels and metabolic activity level of the key drug metabolic enzyme CYP3A1 in rat liver. Methods Wistar rats were randomly divided into control group, 24 h post-radiation group and 72 h post-radiation group. The experimental group was exposed to total body irradiation of single 6 Gy γ-ray. Blood was collected from the orbital venous plexus for blood routine examination and biochemical analysis 24 h and 72 h after irradiation, and liver tissue was prepared for quantifying expression of CYP3A1 mRNA and liver-specific microRNA (miR-122-5p) through RT-PCR. The expression level of CYP3A1 protein was analyzed by Western blot, and the metabolic activity level of CYP3A1 detected by the specific substrate midazolam combined with LC-MS method. Results Com¬pared with the control group, the weights of the rats in the radiation group significantly decreased, and the number of white blood cells was markedly reduced. Simultaneously, the activities of alanine aminotrans-ferase and alkaline phosphatase continuously descended, as well as the levels of total bilirubin and bile acid significantly increased, which indicated that the liver may be damaged after radiation. The relative expression of CYP3A1 mRNA continued to increase significantly 24 h and 72 h after irradiation. CYP3A1 protein expression and metabolic activity levels showed an obvious increasing trend 24 h after irradiation, and rose significantly 72 h after irradiation compared with the control group. At the same time, the expression of miR-122-5p in liver of rats in the 24 h and 72 h post-radiation group continued to decrease rapidly compared with the control group. Conclusions γ-ray radiation may arouse damage effect on liver, which leads to the continuous up-regulation of the mRNA and protein expression levels of the capital metabolic enzyme CYP3A1 in liver tissue, as well as the elevation of the metabolic activity level. The regulatory mechanism might be related to miR-122-5p.

7.
Chinese Pharmacological Bulletin ; (12): 1819-1823, 2023.
Article in Chinese | WPRIM | ID: wpr-1013688

ABSTRACT

Pericytes are essential components of vessel mural cells that function to regulate blood flow, clear or phagocytose debris, and are contractile cells enwrapping capillaries throughout the body. It controls vascular permeability and is involved in the development of blood vessels and is an important regulator and potential drug target of angiogenesis and vascular function. Pericytes are also thought to play a key role in the tumor microenvironment, especially during tumor growth and distal metastasis. Therefore,in this review we discuss the relationship between pericytes involved in tumor angiogenesis and tumor metastasis, as well as the use of targeted pericytes to treat tumors,with a view to providing a basis for subsequent studies.

8.
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery ; (12): 982-988, 2023.
Article in Chinese | WPRIM | ID: wpr-996721

ABSTRACT

@# Objective     To analyze the efficacy of off-pump coronary artery bypass grafting (OPCABG) in elderly patients with coronary artery disease complicated with moderate ischemic mitral regurgitation. Methods     The clinical data of patients aged≥70 years with coronary artery disease complicated with moderate mitral regurgitation, and undergoing OPCABG from January 2009 to January 2020 in Beijing Anzhen Hospital were retrospectively analyzed. The echocardiographic indicators of the patients were compared preoperatively, postoperatively before discharge and during the follow-up. Results     Finally 239 patients were enrolled. There were 136 males and 103 females, aged 74.1±3.2 years. Before postoperative discharge, 49 (20.5%) patients had no mitral regurgitation, 144 (60.3%) mild regurgitation, 46 (19.2%) moderate regurgitation, and 0 severe regurgitation. The area of mitral regurgitation was significantly improved (2.5±1.8 cm2 vs. 5.6±1.0 cm2, P<0.001). There were 10 (4.2%) patients of hospital death, 23 (9.6%) of low cardiac output, 3 (1.3%) of myocardial infarction, and 8 (3.3%) of nervous system injury after operation. As a result, 208 (90.8%) patients were followed up and the mean follow-up time was 3.4 years (range 1-9 years). The cumulative survival rates at postoperative 2, 4, 6, and 8 years were 95.8%, 88.0%, 78.4%, and 73.1%, respectively. Postoperative follow-up showed significant improvements compared with those before surgery in the area of mitral regurgitation, left ventricular ejection fraction, left ventricular end-diastolic and left ventricular end-systolic diameters (all P<0.05). Duirng the follow-up, the major adverse cardiac and cerebrovascular events were all cause death in 22 (10.6%) patients, including cardiac death in 17 (8.2%) patients, myocardial infarction in 7 (3.4%) patients, heart failure in 24 (11.5%) patients, cerebrovascular events in 11 (5.3%) patients, re-hospitalization due to heart disease in 23 (11.1%) patients, and none of the patients with myocardial infarction were revascularized. Conclusion     The mid- and long-term outcomes of OPCABG in the treatment for elderly patients with coronary artery disease complicated with moderate ischemic mitral regurgitation is good.

9.
Chinese Journal of Medical Aesthetics and Cosmetology ; (6): 33-35, 2023.
Article in Chinese | WPRIM | ID: wpr-995898

ABSTRACT

Objective:To explore the method and effect of blepharoplasty of upper eyelid combined with orbicularis oculi myocutaneous flap in repair of the canthus defect after excision of xanthelasma.Methods:According to the relaxation of the upper eyelid skin and the width of the xanthoma to be removed, the double eyelid line and the peeling line were designed individually, and the myocutaneous flap was removed to repair the inner canthus defect. After resection of the xanthelasma, the orbicularis oculi myocutaneous flap was transferred retrogradely to repair the defect, and then the upper eyelid plasty was performed.Results:During the follow-up of 3-12 months after operation, 13 cases (24 eyes) of the transferred flaps in the inner canthus survived well, the incision scars were hidden, and the double eyelids on both sides were naturally symmetrical.Conclusions:The method of upper eyelid plasty combined with the reverse transfer of orbicularis oculi myocutaneous flap provides more options for the treatment of defects after resection of xanthoma of the inner canthus. The flap survives well, improves the shape of the upper eyelid while completely removing the focus, and avoids complications such as postoperative deformation and obvious scars.

10.
Chinese Journal of Urology ; (12): 42-46, 2023.
Article in Chinese | WPRIM | ID: wpr-993969

ABSTRACT

Objective:To compare the efficacy of modified "double flaps" pyeloplasty and traditional dismembered pyeloplasty in the treatment of special types of hydronephrosis with small pelvis and long proximal ureteral stricture in children.Methods:The data of 39 children with special types of hydronephrosis treated in Seventh Medical Center, General Hospital of PLA from June 2018 to June 2019 were retrospectively analyzed. Among them, 33 were boys and 6 were girls. The median age of the patients was 12.0(4.5, 63.5) months. Nine of them had left hydronephrosis and four children had right hydronephrosis. These patients with small pelvis existed the characteristics that the anteroposterior diameter of pelvis was smaller than 2.5 cm and these patients existed the symptom of hematuria, flank pain or recurrent urinary tract infection with the imaging revealing ureteral obstruction. The length of proximal ureteral stenosis ranged from 2.0 to 4.0 cm. Among 39 cases, 19 cases were operated with modified "double flaps" pyeloplasty, which was the modified "double flaps" pyeloplasty group. 20 cases were operated with traditional Anderson-Hynes pyeloplasty, which was traditional pyeloplasty group. The technique of modified "double flaps" pyeloplasty mainly included that the renal pelvis was cut into double flaps, the inferior flap was anastomosed with the spatulated ureter and the superior was covered, so that the length and caliber of the ureter were partial extended. The median age of two groups were 12.0 (6.0, 44.0) months and 12.0 (4.8, 62.8) months respectively, the anterior and posterior diameter of renal pelvis were (2.8±0.8)cm and (2.6±0.6)cm respectively, and split renal function were (36.7±5.1)% and (36.0±6.8)% respectively. There were no statistically significant differences in above parameters between the two groups( P>0.05). The clinical efficacy of the two groups were compared by collecting and comparing the operation related data and postoperative follow-up data. Results:The operation of 39 children in this study was successfully completed without conversion to open surgery.The operation time of "double flap" pyeloplasty group and traditional pyeloplasty group were (142.6±9.6) min and (124.5±8.6) min respectively, and the intraoperative anastomosis time were (56.1±7.2) min and (47.6±4.8) min respectively. There were significant difference in operation time and intraoperative anastomosis time between the two groups( P<0.05). Thirty-nine children were followed up normally without loss. The mean follow-up time was (27.7±2.5) months after surgery. In the "double flaps" pyeloplasty group, 2 cases suffered with fever who were diagnosed as urinary tract infection and improved after antibiotic treatment. In the traditional pyeloplasty group, 2 cases suffered with fever who were diagnosed as urinary tract infection and improved after antibiotic treatment. Two children had flank pain during follow-up to more than one year and the examination revealed that the anteroposterior diameter of the renal pelvis gradually increased. So surgery were performed again and the two children recovered. There were no significant differences in complication rate (2/19 and 4/20) and short-term surgical success rate(19/19 and 18/20) between the two groups ( P>0.05). Conclusions:The operation time and anastomosis time of the modified "double flap" technique for treating hydronephrosis are longer than those of the traditional method. But in the treatment of special types of hydronephrosis with small renal pelvis or long proximal ureteral stricture, it may have application prospects in reducing complications.

11.
Chinese Journal of Health Management ; (6): 222-227, 2023.
Article in Chinese | WPRIM | ID: wpr-993659

ABSTRACT

Objective:To analyze the current situation and effect of mobile applications for physical activity management in China.Methods:In this systematic review, search terms such as “exercise”, “sports”, “exercise”, “physical activity” and “mobile health” were used to retrieve the applications for physical activity management in the IOS Application Store developed in mainland of China. Then the Mobile Application Rating Scale (MARS) and the Behavior Change Technology (BCT) were used to evaluate the quality of the applications and analyze the behavior change technologies that applications adopted. Moreover, a systematic review of relevant experimental studies was applied to summarize the effectiveness of the applications in promoting physical activity.Results:Total of 141 applications were included in this analysis, and 58 behavior change techniques in 16 categories were applied. The most frequently used 4 categories of BCT were behavior comparison (67.4%), repetition and substitution (67.4%), knowledge construction (67.4%), feedback and monitoring (58.2%). The overall objective quality score of MARS was 3.17±0.42, of which, the score of engagements, functionality, aesthetic design and information quality was 2.67±0.73, 3.75±0.54, 3.44±0.59 and 2.83±0.55, respectively. The number of BCT adopted in each application had a linear relationship with the application′s objective quality score in MARS ( B=0.023, 95% CI: 0.018-0.029) and user rating ( B=0.040, 95% CI: 0.019-0.060) (both P<0.001). A total of 4 articles were included to test the effects of techniques (i.e., physical activity behavior monitoring, physical activity planning and physical activity guidance) in 4 applications. The overall quality of the literatures was good. The main outcomes were physical activity duration (3 articles) and step count (1 article). Compared with those who did not use the application or only used the accelerometer to monitor physical activity, subjects who used the application to monitor physical activity had significant longer physical activity durations and step count (both P<0.05). Conclusions:The number, function and technology of physical activity management applications in China are abundant, but the quality of the applications is moderate. The number of BCT is an important factor in evaluating application′s quality, but its effect on behavior change is still not clear.

12.
China Pharmacy ; (12): 2233-2237, 2023.
Article in Chinese | WPRIM | ID: wpr-988783

ABSTRACT

OBJECTIVE To establish a method for concentration determination of caffeine and its three metabolites, theophylline, paraxanthine and theobromine in urine, and apply it in clinical practice. METHODS Using caffeine-13C3-d3 as internal standard (IS), and the urine samples were protein precipitated with acetonitrile; HPLC-MS/MS method was adopted to determine the concentrations of caffeine and its three metabolites. The determination was performed on Waters ACQUITY UPLC® BEH HILIC column with mobile phase consisting of 60 mmol/L ammonium acetate (A)-acetonitrile (B) (gradient elution) at the flow rate of 0.5 mL/min. The column temperature was set at 38 ℃ , and the sample size was 2 μL. The electrospray ionization detection was operated in a positive mode by multiple reaction monitoring. The detection ions for quantitative analysis were m/z 195.1→110.0 for caffeine, m/z 181.1→124.0 for theophylline, m/z 181.1→124.0 for paraxanthine, m/z 181.1→138.0 for theobromine, and m/z 198.1→ 140.1 for IS. The above method was used to determine the concentrations of caffeine and its three metabolites in the urine of 19 infants with apnea of prematurity (AOP). RESULTS The linear ranges of mass concentration of caffeine, theophylline, paraxanthin and theobromine were 0.200-200, 0.050-50.0,0.050 0-50.0, and 0.100-100 μg/mL, respectively. The lower limits of quantification were 0.200, 0.050, 0.050 and 0.100 μg/mL (r>0.990), respectively. RSDs of intra-day and intra- day precision were not above 10.37%, and matrix factors were 85.68%-109.90%; extraction recoveries were 93.53%-109.40% (RSD≤15%), and RSDs of stability tests were all lower than 15%. The concentrations of caffeine and its three metabolites in the urine of 19 cases were (27.346±7.951), (0.351±0.223), (0.428±0.395) and (0.472±0.374) μg/mL, respectively. CONCLUSIONS The established HPLC-MS/MS method is simple, sensitive and can be used for the determination of caffeine and its three metabolites in urine samples of AOP.

13.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 870-877, 2023.
Article in Chinese | WPRIM | ID: wpr-988736

ABSTRACT

ObjectiveTo develop a deep learning system for early ultrasound screening of developmental dysplasia of the hip (DDH), a new smart-hip ultrasound technique (S-hip), and to validate its clinical application. MethodsWe selected 11,100 annotated and reviewed coronal ultrasound images of infant hips between November 2021 and August 2022, 8,100 of which were used for the training set and 3,000 for the test set, to build a S-hip deep learning system. To verify the consistency between the automated measurement by S-hip and the manual measurements by sonographers, 174 standard coronal ultrasound images of 87 infants' bilateral hips were acquired, then α angle, β angle and femoral head coverage (FHC) were measured by S-hip, an ultrasound expert and a resident. The measurement data and the time required for the measurements were recorded and statistically analyzed. Another 100 standard coronal ultrasound images of the hips were randomly selected and measured twice respectively by the ultrasound expert and resident to assess the intra-sonographer repeatability. ResultsThe intraclass correlation coefficient (ICC) (95% CI) values of α angle, β angle and FHC results measured by S-hip and ultrasound expert were 0.799 (0.738, 0.847), 0.798 (0.737, 0.846) and 0.934 (0.954, 0.975), respectively. Those values measured by the ultrasound expert and resident were 0.725 (0.645, 0.789), 0.674 (0.583, 0.748) and 0.931 (0.908, 0.949), respectively. The mean absolute errors (MAE) of α angle, β angle and FHC results between measurements by S-hip and ultrasound expert were 2.69 °, 4.43 ° and 2.47%, respectively. The time required for measurements by S-hip, ultrasound expert and resident was (1.59±0.36) s, (18.76±2.23) s and (19.45±2.76) s, respectively. The automated measurement by S-hip cost much shorter time than the manual measurements by sonographers and the difference was statistically significant (P<0.001). The ICC (95% CI) values of α angle, β angle and FHC results between two measurements by the ultrasound expert were 0.943 (0.916, 0.961), 0.959 (0.940, 0.972), and 0.981 (0.971, 0.987), respectively. Those values by the ultrasound resident were 0.884 (0.833, 0.921), 0.921 (0.884, 0.946), and 0.962 (0.944, 0.974). ConclusionThe S-hip based on a deep learning system is a highly reliable automated technique to accurately measure α angle, β angle and FHC. Compared with ultrasound residents, S-hip allows for a more simplified and significantly quicker measurement, which may enhance the widespread use of hip ultrasound screening in infants.

14.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 768-775, 2023.
Article in Chinese | WPRIM | ID: wpr-988722

ABSTRACT

ObjectiveTo investigate the effect of liquiritigenin (LG) on intestinal flora in menopausal APP/PS1 mice. MethodsA total of forty 3-month-old female APP/PS1 mice were randomly divided into sham surgery group (n=20) and ovariectomy group (n=20). Seven days after surgery, the ovariectomy group was randomly divided into ovariectomy control group (OVX, n=10), ovariectomy + liquiritigenin treatment group (OVX + LG, n=10), and the sham surgery group was randomly divided into liquiritigenin treatment group (LG, n=10) and reagent control group (Sham, n=10), and ten C57BL/6J mice were taken as WT group. The dose of LG group and OVX + LG group was 30 mg•kg-1•d-1. After 90 days of drug treatment, fecal samples were gathered, genomes were extracted, and intestinal flora were analyzed by 16S rDNA Amplicon Sequencing. Morris water maze was performed to evaluate learning and memory abilities of mice. Immunofluorescence was used to observe the deposition of senile plaques (SP) in the brain of mice. ResultsThe results of water maze showed that LG significantly improved the learning memory ability of APP/PS1 mice with/without OVX (P<0.05), and reduced the number of SPs in the brain of APP/PS1 mice with/without OVX, and the differences were statistically significant (P<0.000 1). 16s rDNA sequencing analysis of the relative abundance of gut microbiota proved that LG treatment significantly increased the relative abundance of Firmicutes and Lactobacillus (P<0.05) and reduced the relative abundance of harmful bacteria belong to Bacteroidetes (P<0.05) in APP/PS1 mice intestines with/without menopause. After LG treatment, the relative abundance of Allobaculun elevated in the intestines of APP/PS1 mice, while declined in the intestines of menopausal APP/PS1 mice, but the difference was not statistically significant. LEfSe analysis revealed the bacteria with the most differential abundance of the gut microbiota of WT mice were Firmicutes, Bacillus, and Lactobacillales (P<0.05); Lactobacillus reuteri had a greater influence on the LG group (P<0.05); Bacteroidia, Bacteroidales and Bacteroides gathered in the intestines of mice in the Sham group (P<0.05). Firmicutes and Allobaculum were the dominant in the WT group (P<0.05); Bacteroides, Bacteroidia and Bacteroidales were more abundant in the Sham group(P<0.05); Bacterroidaceae and Bacteroides had the most differential abundances in the OVX group (P<0.05); Lactobacillaceae and Lactobacillus were more abundant in the intestines in the OVX + LG group (P<0.05). ConclusionLG could improve the ratio of beneficial and harmful bacteria in the intestines of APP/PS1 mice before and after menopause. Liquiritigenin treatment showed consistent variations in intestinal flora in APP/PS1 mice with or without ovariectomy. It is presumed that menopausal APP/PS1 mice have lipid metabolism disorders which requires further study.

15.
China Tropical Medicine ; (12): 186-2023.
Article in Chinese | WPRIM | ID: wpr-979614

ABSTRACT

@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

16.
Journal of Experimental Hematology ; (6): 135-140, 2023.
Article in Chinese | WPRIM | ID: wpr-971115

ABSTRACT

OBJECTIVE@#To analyze the clinical features and prognosis of patients with Castleman's disease (CD) and improve the diagnosis and treatment of CD.@*METHODS@#Clinical data of patients diagnosed with CD by pathological biopsy in Gansu Provincial Hospital from January 2009 to November 2020 were retrospectively analyzed. According to clinical classification, the patients were divided into two groups: UCD (unicentric CD) group (n=20) and MCD (multicentric CD) group (n=9). The clinical manifestations, laboratory examination, treatment regimens, pathological examination and follow-up data were statistically analyzed.@*RESULTS@#There were no significant differences in average age and gender ratio between UCD group and MCD group. In UCD patients, 80.0% were hyaline vascular type, and 20.0% were plasma cell type. In MCD patients, 33.3% were hyaline vascular type, 55.6% were plasma cell type, and 11.1% were mixed type. There was significant difference in pathological classification between the two groups (P=0.039). The UCD patients usually presented asymptomatic single lymph node enlargement with mild clinical symptoms, while the MCD patients were characterized by multiple superficial and deep lymph node enlargement throughout the body. The incidences of asthenia, splenomegaly, serous effusion in MCD group were higher than those in UCD group (P<0.05). Meanwhile, the incidences of anemia, hypoproteinemia, increased ESR, elevated serum globulin and elevated β2-microglobulin were significantly higher than those in UCD group too (P<0.05). There was no significant difference in the incidences of abnormal WBC, PLT and elevated LDH between the two groups (P>0.05). Among 20 patients with UCD, 13 cases reached complete remission (CR), 1 case achieved partial remission (PR). Among 9 patients with MCD, 3 cases received CR and 4 cases received PR.@*CONCLUSION@#Patients with CD requires pathological examination for diagnosis. Patients with UCD show mild clinical symptoms, good surgical treatment effect and good prognosis. Patients with MCD have diversified clinical manifestations and relatively poor prognosis, and these patients require comprehensive treatment.


Subject(s)
Humans , Castleman Disease/therapy , Retrospective Studies , Prognosis , Splenomegaly , Anemia
17.
Protein & Cell ; (12): 477-496, 2023.
Article in English | WPRIM | ID: wpr-982528

ABSTRACT

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.


Subject(s)
Male , Mice , Animals , Cellular Reprogramming/genetics , Tetraploidy , Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/metabolism , DNA Methylation , Spermatogonia/metabolism , Germ Cells/metabolism
18.
Chinese journal of integrative medicine ; (12): 299-307, 2023.
Article in English | WPRIM | ID: wpr-982286

ABSTRACT

OBJECTIVE@#To evaluate the efficacy and safety of Jianpi Jieyu Decoction (JJD) for treating patients with mild-to-moderate depression of Xin (Heart)-Pi (Spleen) deficiency (XPD) syndrome.@*METHODS@#In this multi-center, randomized, controlled study, 140 patients with mild-to-moderate depression of XPD syndrome were included from Xiyuan Hospital of China Academy of Chinese Medical Sciences and Botou Hospital of Traditional Chinese Medicine from December 2017 to December 2019. They were randomly divided into JJD group and paroxetine group by using a random number table, with 70 cases in each group. The patients in the JJD group were given JJD one dose per day (twice daily at morning and evening, 100 mL each time), and the patients in the paroxetine group were given paroxetine (10 mg/d in week 1; 20 mg/d in weeks 2-6), both orally administration for a total of 6 weeks. The primary outcome was the change of 17-item Hamilton Depression Rating Scale (HAMD-17) score at week 6 from baseline. The secondary outcomes included the Hamilton Anxiety Scale (HAMA) score, Traditional Chinese Medicine Symptom Scale (TCMSS), and Clinlcal Global Impression (CGI) scores at the 2nd, 4th, and 6th weekends of treatment, HAMD-17 response (defined as a reduction in score of >50%) and HAMD-17 remission (defined as a score of ⩽7) at the end of the 6th week of treatment. Adverse events (AEs) were also recorded.@*RESULTS@#From baseline to week 6, the HAMD-17 scores decreased 10.2 ± 4.0 and 9.1 ± 4.9 points in the JJD and paroxetine groups, respectively (P=0.689). The HAMD-17 response occurred in 60% of patients in the JJD group and in 50% of those in the paroxetine group (P=0.292); HAMD-17 remission occurred in 45.7% and 30% of patients, respectively (P=0.128). The differences of CGI scores at the 6th week were not statistically significant (P>0.05). There were significant differences in HAMD-17 scores between the two groups at 2nd and 4th week (P=0.001 and P=0.014). The HAMA scores declined 8.1 ± 3.0 and 6.9 ± 4.3 points from baseline to week 6 in the JJD and paroxetine groups, respectively (P=0.905 between groups). At 4th week of treatment, there was a significant difference in HAMA between the two groups (P=0.037). TCMSS decreased 11.4 ± 5.1, and 10.1 ± 6.8 points in the JJD and paroxetine groups, respectively (P=0.080 between groups). At the 6th week, the incidence of AEs in the JJD group was significantly lower than that in the paroxetine group (7.14% vs. 22.86%, P<0.05).@*CONCLUSION@#Compared with paroxetine, JJD was associated with a significantly lower incidence of AEs in patients with mild-to-moderate depression of XPD syndrome, with no difference in efficacy at 6 weeks. (Trial registration No. ChiCTR2000040922).


Subject(s)
Humans , Paroxetine/adverse effects , Spleen , Anxiety , Syndrome , Medicine, Chinese Traditional , Treatment Outcome , Double-Blind Method
19.
China Journal of Chinese Materia Medica ; (24): 3156-3161, 2023.
Article in Chinese | WPRIM | ID: wpr-981451

ABSTRACT

Baby Boom(BBM) gene is a key regulatory factor in embryonic development and regeneration, cell proliferation, callus growth, and differentiation promotion. Since the genetic transformation system of Panax quinquefolius is unstable with low efficiency and long period, this study attempted to transfer BBM gene of Zea mays to P. quinquefolius callus by gene gunship to investigate its effect on the callus growth and ginsenoside content, laying a foundation for establishing efficient genetic transformation system of P. quinquefolius. Four transgenic callus of P. quinquefolius with different transformation events were obtained by screening for glufosinate ammonium resistance and molecular identification by PCR. The growth state and growth rate of wild-type and transgenic callus were compared in the same growth period. The content of ginsenoside in transgenic callus was determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The results showed that transgenic callus growth rate was significantly higher than that of wild-type callus. In addition, the content of ginsenoside Rb_1, Rg_1, Ro, and Re was significantly higher than that in wild-type callus. The paper preliminarily proved the function of BBM gene in promoting growth rate and increasing ginsenoside content, which provided a scientific basis to establish a stable and efficient genetic transformation system for Panax plants in the future.


Subject(s)
Female , Pregnancy , Humans , Ginsenosides , Panax/genetics , Chromatography, Liquid , Tandem Mass Spectrometry , Cell Proliferation
20.
China Journal of Chinese Materia Medica ; (24): 3149-3155, 2023.
Article in Chinese | WPRIM | ID: wpr-981450

ABSTRACT

This study explored the preservation effect of strigolactone analogs on Gastrodia elata tubers and screened out the suitable preservation measures of G. elata to provide a safer and more effective method for its storage and preservation. Fresh G. elata tubers were treated with 7FGR24, 2,4-D isooctyl ester, and maleic hydrazide, respectively. The growth of flower buds, the activities of CAT, and MDA, and the content of gastrodin and p-hydroxybenzyl alcohol were measured to compare the effects of different compounds on the storage and preservation of G. elata. The effects of different storage temperatures on the preservation of 7FGR24 were compared and analyzed. The gibberellin signal transduction receptor gene GeGID1 was cloned, and the effect of 7FGR24 on the expression level of GeGID1 was analyzed by quantitative polymerase chain reaction(qPCR). The toxicity of the G. elata preservative 7FGR24 was analyzed by intragastric administration in mice to evaluate its safety. The results showed that compared with 2,4-D isooctyl ester and maleic hydrazide, 7FGR24 treatment had a significant inhibitory effect on the growth of G. elata flower buds, and the CAT enzyme activity of G. elata was the highest, indicating that its preservation effect was stronger. Different storage temperatures had different effects on the preservation of G. elata, and the preservation effect was the strongest at 5 ℃. The open reading frame(ORF) of GeGID1 gene was 936 bp in length, and its expression level was significantly down-regulated after 7FGR24 treatment, indicating that 7FGR24 may inhibit the growth of flower buds by inhibiting the gibberellin signal of G. elata, thereby exerting a fresh-keeping effect. Feeding preservative 7FGR24 had no significant effect on the behavior and physiology of mice, indicating that it had no obvious toxicity. This study explored the application of the strigolactone analog 7FGR24 in the storage and preservation of G. elata and preliminarily established a method for the storage and preservation of G. elata, laying a foundation for the molecular mechanism of 7FGR24 in the storage and preservation of G. elata.


Subject(s)
Animals , Mice , Gastrodia , Gibberellins , Maleic Hydrazide , Esters
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