Application Progress of Electrochemical Methods in Quality Control of Traditional Chinese Medicine / 分析化学

The quality control of traditional Chinese medicine(TCM)is the core issue to ensure the modernization,industrialization and internationalization of TCM.Compared with other detection methods,electrochemical analysis method has many advantages such as high sensitivity,fast detection speed and low cost,making it an important means of quality control for TCM and having broad development prospects.This article reviewed the research progress of electrochemical methods in quality control of TCM in recent years,discussed the application of electrochemical fingerprinting technique in identification of TCM,and comprehensively summarized the application of electrochemical technology in analyzing effective components and harmful substances in TCM,including flavonoids,alkaloids,quinones,glycosides,heavy metals and pesticide residues.Finally,the development prospects of electrochemical methods in the field of quality control of TCM were discussed.

Mechanism of Anti-inflammatory Effects of Bupi Yichang Pills on Inhibiting Glycolytic Metabolic Pathway in Mice with Experimental Colitis / 中药新药与临床药理

Objective To investigate the anti-inflammatory effects of Bupi Yichang Pills on mice with experimental colitis and its potential mechanism of action.Methods Dextran sulfate sodium(DSS)was used to model the experimental colitis,and low-,medium-and high-doses of Bupi Yichang Pills(1.5,3.0,6.0 g·kg-1·d-1)and Mesalazine(300 mg·kg-1·d-1)were fed at the same time.Mice were observed for general behavior and weighed.Hematoxylin-eosin staining was used to observe the pathological injury of colonic tissues.qPCR and ELISA were used to detect the levels of inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10,IL-35 and TGF-β1),qPCR and Western Blot were used to detect the mRNA and protein levels of glucose transporters and glycolytic kinases.Results Low-,medium-and high-doses of Bupi Yichang Pills significantly down-regulated disease activity index in colitis mice(P＜0.05,P＜0.01).The body mass and colon length were significantly increased,while colon mass,colon mass index and unit colon mass index were significantly reduced(P＜0.05,P＜0.01),and ulcer formation and inflammatory cell infiltration in colonic tissue were significantly improved.In addition,medium-and high-doses of Bupi Yichang Pills significantly down-regulated the mRNA levels and concentrations of pro-inflammatory cytokines including TNF-α,IL-1β and IL-6(P＜0.01),while significantly up-regulated the mRNA levels and concentrations of anti-inflammatory cytokines such as IL-10,IL-35 and TGF-β1(P＜0.01).We further found that high-dose of Bupi Yichang Pills significantly down-regulated the mRNA and protein expressions of glucose transporters(Glut1,Glut2,Glut4)and glycolytic kinases(HK2,Aldolase A,PKM2)in colonic tissue(P＜0.01).Conclusions Bupi Yichang Pills effectively alleviates DSS-induced experimental colitis,and its specific mechanism of action is related to the improvement of glycolytic metabolic pathways and the regulation of inflammatory cytokine expression.

Mechanism of Active Ingredients of Astragalus Membranacus in Treating Heart Failure： A Review / 中国实验方剂学杂志

Heart failure is one of the main cardiovascular system diseases at present， and it is a clinical syndrome caused by changes in cardiac structure and function， resulting in impaired ejection function or ventricular filling. Therefore， heart failure has become the most important cardiovascular disease in the 21st century. In recent years， the incidence of heart failure is increasing， and the survival rate of patients with heart failure is very low. Traditional Chinese medicine has rich experience in preventing and treating heart failure. With the modernization of traditional Chinese medicine， more and more attention has been paid to the research， development， and application of active ingredients in traditional Chinese medicine. Traditional Chinese medicine has unique advantages in improving the heart function of patients with heart failure by treating multiple targets and multiple pathways through syndrome differentiation. Astragalus membranacus， a traditional Chinese medicine， is a kind of medicine that benefits Qi and blood circulation and removes evil spirits. It has the functions of improving myocardial energy metabolism and hemodynamics， protecting myocardial muscle， and promoting angiogenesis. Astragalus membranaceus is often used to treat patients with heart failure， yielding remarkable results. In recent years， it has been found that astragaloside， Astragalus polysaccharide， quercetin， calyx isoflavones， and other main active ingredients of Astragalus membranacus can improve cardiac function and treat heart failure by inhibiting inflammatory response， myocardial apoptosis， and myocardial fibrosis. This paper reviewed the research progress of the action and mechanism of the active ingredients of Astragalus membranacus in the treatment of heart failure by studying relevant literature， with a view to providing a reference for its further research， development， and application in the prevention and treatment of heart failure.

Effect of Yixintai on Mitochondrial Fission Proteins Fis1 and Mff in Rat Model of Chronic Heart Failure / 中国实验方剂学杂志

ObjectiveTo study the effect and mechanism of Yixintai on mitochondrial fission proteins in the rat model of chronic heart failure. MethodTen of 60 SD rats were randomly selected as the sham operation group， and the remaining 50 rats were subjected to ligation of the left anterior descending coronary artery for the modeling of heart failure post myocardial infarction. The successfully modeled rats were randomized into model， low-， medium-， and high-dose （1.4， 2.8， and 5.6 g·kg-1， respectively） Yixintai， and trimetazidine （10 mg·kg-1） groups. The rats were administrated with corresponding doses of drugs by gavage， and the rats in the model group and sham operation group were given an equal volume of normal saline by gavage for 28 consecutive days. Enzyme-linked immunosorbent assay （ELISA） was then employed to measure the levels of amino-terminal pro-B-type natriuretic peptide （NT-pro BNP）， B-type natriuretic peptide （BNP）， and adenosine triphosphate （ATP） in the serum. Color Doppler ultrasound imaging was conducted to examine the cardiac function indicators. Hematoxylin-eosin staining and Masson staining were conducted to observe the pathological changes in the heart， and Image J was used to calculate collagen volume fraction （CVF）. Transmission electron microscopy was employed to observe the ultrastructural changes of myocardial cells. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to measure the apoptosis rate of myocardial cells. Western blot was employed to determine the protein levels of mitochondrial fission protein 1 （Fis1） and mitochondrial fission factor （Mff） in the outer mitochondrial membrane of the myocardial tissue. ResultCompared with the sham operation group， the model group showed elevated levels of NT-pro BNP and BNP in the serum， decreased ATP content， left ventricular ejection fraction （LVEF）， and left ventricular fraction shortening （LVFS）， increased left ventricular end-diastolic diameter （LVIDd） and left ventricular end-systolic diameter （LVIDs）， disarrangement of myocardial cells， inflammatory cell infiltration， increased collagen fibers and CVF， damaged myocardium and mitochondria， and increased apoptosis rate of myocardial cells， and up-regulated expression of Fis1 and Mff in the cardiac tissue （P<0.01）. Compared with the model group， different doses of Yixintai and trimetazidine lowered the serum levels of NT-pro BNP and BNP （P<0.05）， increased the ATP content （P<0.05）， increased LVEF and LVFS （P<0.01）， decreased LVIDd and LVIDs （P<0.01）. Moreover， the drugs alleviated the myocardial inflammatory damage and fibrosis， reduced CVF （P<0.01）， repaired the myocardial mitochondrial structure， and decreased the apoptosis rate of myocardial cells （P<0.01）. Medium- and high-dose Yixintai and trimetazidine down-regulated the expression of Fis1 and Mff in the myocardial tissue （P<0.05）. ConclusionYixintai can improve mitochondrial structure， reduce myocardial cell apoptosis， and improve cardiac function by inhibiting the expression of Fis1 and Mff in the myocardial tissue.

The Path Analysis of Integrating Sun Simiao’s Medical Ethics Thought into the Humanistic Spirit of Medical Students / 中国医学伦理学

Medicine is the subject of studying human and its inherent humanistic attribute endows medicine with the temperature it deserves. However, with the continuous improvement of medical testing technology and treatment technology, medical workers pay more attention to diseases and ignore humanistic care, which has become an important factor in the aggravation of patients’ burden and the tension relationship between doctors and patients. There are many virtues in Chinese traditional culture. Sun Simiao’s medical ethics thought of "great doctor with professionalism and sincerity" embodies the core value of the humanistic spirit of Chinese traditional medicine. In his medical ethics thought, the moral and ethics of "great doctor with professionalism and sincerity", the medical practice attitude of "being cautious and diligent", the values of "benevolence for the world" and the professional conduct of "honesty and truth" still have strong practical significance at present. It is the good material to cultivate the humanistic spirit of medical students. This is of great value to integrate Sun Simiao’s medical ethics thought into cultivate medical students with both "professionalism" and "sincerity", practice the original intention of medical and health undertakings, and carry forward cultural self-confidence.

The ethical review and reflection on in-utero pediatrics / 中国医学伦理学

In-utero pediatrics is a fetal general medicine for perinatal medicine redevelopment,with the main purpose of preventing and treating fetal diseases,which studies the early prevention,screening,diagnosis,and treatment of diseases from gametes,fertilized eggs,and embryos sequential to children,adolescents,adolescence,and even the entire life cycle.Medical ethics provides strong support for the normative development of this discipline.This paper summarized the formation and development of in-utero pediatrics,analyzed the vulnerability of service subjects within in-utero pediatrics,and sorted out their ethical issues in the prevention and control of birth defects,fetal intrauterine diagnosis and treatment,as well as multidisciplinary collaborative diagnosis and treatment.It was proposed that in-utero pediatrics should follow the medical principle of maternal and fetal interests first,the principle of respect,and the principle of no harm.Finally,suggestions for ethical review of clinical and research projects on in-utero pediatrics were proposed,including strengthening the advisory service role of the ethics committee in clinical practice,timely launching the guidelines of ethical review for clinical research,and enhancing the ethical awareness of medical staff.

Discussion on the Effects of Mitochondrial Homeostasis in Heart Failure Based on"Yin-yang Theory" / 中国中医药信息杂志

Heart failure is a group of complex clinical syndromes in the middle and late stages of cardiovascular diseases.Mitochondrial homeostasis imbalance is one of the pathological mechanisms in the occurrence and development of heart failure.This article revolved around the"yin-yang theory"in TCM and explained the pathological mechanism of heart failure through mitochondrial homeostasis.Heart failure is the syndrome of deficiency in nature and excess in superficiality fundamental.Its basic pathogenesis is"yang deficiency and yin excess".Based on the deficiency of heart yang qi and the stagnation of yin pathogens,the combination of deficiency and excess runs through the entire disease.Mitochondrial homeostasis imbalance is a manifestation of yin-yang imbalance at the cellular micro level,mainly manifested as inhibition of mitochondrial biosynthesis,mitochondrial dynamics imbalance,mitophagy disorder,etc.,which affects mitochondrial structure and function and leads to abnormal myocardial energy metabolism.Therefore,based on the"yin-yang theory",the basic treatment method is to"tonify deficiency and damage excess"to regulate mitochondrial biosynthesis,mitochondrial dynamics,and mitophagy,thereby maintaining mitochondrial homeostasis and improving myocardial energy metabolism,which is of great significance for the prevention and treatment of heart failure.

Effect of Erxian Decoction-containing serum on H_2O_2-induced proliferation and osteogenic differentiation of MC3T3-E1 cells via BK channels / 中国中药杂志

This study aimed to investigate the effects of Erxian Decoction(EXD)-containing serum on the proliferation and osteogenic differentiation of MC3T3-E1 cells under oxidative stress through BK channels. The oxidative stress model was induced in MC3T3-E1 cells by H_2O_2, and 3 mmol·L~(-1) tetraethylammonium(TEA) chloride was used to block the BK channels in MC3T3-E1 cells. MC3T3-E1 cells were divided into a control group, a model group, an EXD group, a TEA group, and a TEA+EXD group. After MC3T3-E1 cells were treated with corresponding drugs for 2 days, 700 μmol·L~(-1) H_2O_2 was added for treatment for another 2 hours. CCK-8 assay was used to detect cell proliferation activity. The alkaline phosphatase(ALP) assay kit was used to detect the ALP activity of cells. Western blot and real-time fluorescence-based quantitative PCR(RT-qPCR) were used to detect protein and mRNA expression, respectively. Alizarin red staining was used to detect the mineralization area of osteoblasts. The results showed that compared with the control group, the model group showed significantly blunted cell proliferation activity and ALP activity, reduced expression of BK channel α subunit(BKα), collagen Ⅰ(COL1), bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG), and phosphorylated Akt, decreased mRNA expression levels of Runt-related transcription factor 2(RUNX2), BMP2, and OPG, and declining area of calcium nodules. EXD-containing serum could significantly potentiate the cell proliferation activity and ALP activity, up-regulate the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt, and forkhead box protein O1(FoxO1), promote the mRNA expression of RUNX2, BMP2, and OPG, and enlarge the area of calcium nodules. However, BK channel blockage by TEA reversed the effects of EXD-containing serum in promoting the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt and FoxO1, increasing the mRNA expression of RUNX2, BMP2, and OPG, and enlarging the area of calcium nodules. EXD-containing serum could improve the proliferation activity, osteogenic differentiation, and mineralization ability of MC3T3-E1 cells under oxidative stress, which might be related to the regulation of BK channels and downstream Akt/FoxO1 signaling pathway.

The role of Keap1/Nrf2/HO-1 signal pathway in liver injury induced by rare earth neodymium oxide in mice / 中华劳动卫生职业病杂志

Objective: To investigate the role of Keap1/Nrf2/HO-1 signaling pathway in liver injury induced by neodymium oxide (Nd(2)O(3)) in mice. Methods: In March 2021, forty-eight SPF grade healthy male C57BL/6J mice were randomly divided into control group (0.9% NaCl), low dose group (62.5 mg/ml Nd(2)O(3)), medium dose group (125.0 mg/ml Nd(2)O(3)), and high dose group (250.0 mg/ml Nd(2)O(3)), each group consisted of 12 animals. The infected groups were treated with Nd(2)O(3) suspension by non-exposed tracheal drip and were killed 35 days after dust exposure. The liver weight of each group was weighed and the organ coefficient was calculated. The content of Nd(3+) in liver tissue was detected by inductively coupled plasma mass spectrometry (ICP-MS). HE staining and immunofluorescence was used to observe the changes of inflammation and nuclear entry. The mRNA expression levels of Keap1, Nrf2 and HO-1 in mice liver tissue were detected by qRT-PCR. Western blotting was used to detect the protein expression levels of Keap1 and HO-1. The contents of catalase (CAT), glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) were detected by colorimetric method. The contents of interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were determined by ELISA. The data was expressed in Mean±SD. Two-independent sample t-test was used for inter-group comparison, and one-way analysis of variance was used for multi-group comparison. Results: Compared with the control group, the liver organ coefficient of mice in medium and high dose groups were increased, and the Nd(3+) accumulation in liver of mice in all dose groups were significantly increased (P<0.05). Pathology showed that the structure of liver lobules in the high dose group was slightly disordered, the liver cells showed balloon-like lesions, the arrangement of liver cell cords was disordered, and the inflammatory exudation was obvious. Compared with the control group, the levels of IL-1β and IL-6 in liver tissue of mice in all dose groups were increased, and the levels of TNF-α in liver tissue of mice in high dose group were increased (P<0.05). Compared with the control group, the mRNA and protein expression levels of Keap1 in high dose group were significantly decreased, while the mRNA expression level of Nrf2, the mRNA and protein expression levels of HO-1 were significantly increased (P<0.05), and Nrf2 was successfully activated into the nucleus. Compared with the control group, the activities of CAT, GSH-Px and T-SOD in high dose group were significantly decreased (P<0.05) . Conclusion: A large amount of Nd(2)O(3) accumulates in the liver of male mice, which may lead to oxidative stress and inflammatory response through activation of Keap1/Nrf2/HO-1 signal pathway. It is suggested that Keap1/Nrf2/HO-1 signal pathway may be one of the mechanisms of Nd(2)O(3) expose-induced liver injury in mice.

Huazhi Rougan Granules attenuates steatosis in cell model of nonalcoholic fatty liver disease by inducing autophagy / 中国中药杂志

To investigate the effect of Huazhi Rougan Granules(HZRG) on autophagy in a steatotic hepatocyte model of free fatty acid(FFA)-induced nonalcoholic fatty liver disease(NAFLD) and explore the possible mechanism. FFA solution prepared by mixing palmitic acid(PA) and oleic acid(OA) at the ratio of 1∶2 was used to induce hepatic steatosis in L02 cells after 24 h treatment, and an in vitro NAFLD cell model was established. After termination of incubation, cell counting kit-8(CCK-8) assay was performed to detect the cell viability; Oil red O staining was employed to detect the intracellular lipid accumulation; enzyme-linked immunosorbnent assay(ELISA) was performed to measure the level of triglyceride(TG); to monitor autophagy in L02 cells, transmission electron microscopy(TEM) was used to observe the autophagosomes; LysoBrite Red was used to detect the pH change in lysosome; transfection with mRFP-GFP-LC3 adenovirus was conducted to observe the autophagic flux; Western blot was performed to determine the expression of autophagy marker LC3B-Ⅰ/LC3B-Ⅱ, autophagy substrate p62 and silent information regulator 1(SIRT1)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK) signaling pathway. NAFLD cell model was successfully induced by FFA at 0.2 mmol·L~(-1) PA and 0.4 mmol·L~(-1) OA. HZRG reduced the TG level(P<0.05, P<0.01) and the lipid accumulation of FFA-induced L02 cells, while elevated the number of autophagosomes and autophagolysosomes to generate autophagic flux. It also affected the functions of lysosomes by regulating their pH. Additionally, HZRG up-regulated the expression of LC3B-Ⅱ/LC3B-Ⅰ, SIRT1, p-AMPK and phospho-protein kinase A(p-PKA)(P<0.05, P<0.01), while down-regulated the expression of p62(P<0.01). Furthermore, 3-methyladenine(3-MA) or chloroquine(CQ) treatment obviously inhibited the above effects of HZRG. HZRG prevented FFA-induced steatosis in L02 cells, and its mechanism might be related to promoting autophagy and regulating SIRT1/AMPK signaling pathway.

Artificial intelligence cell image analysis technology can improve the accuracy of bone marrow cells / 中华检验医学杂志

Objective:To evaluate the screening efficacy of AI for bone marrow cell morphology.Method:Bone marrow specimens of patients attending the Second Hospital of Hebei Medical University from December 1,2019 to December 21,2020;(1) Selected from one hundred bone marrow specimens, The cases included chronic myeloid cell leukemia ( n=23), myelodysplastic syndrome ( n=4), chronic lymphocytic leukemia ( n=4), multiple myeloma ( n=5), 7 acute leukemia ( n=7), chronic anemia ( n=32), infection ( n=6) and healthy control ( n=15). Including 45 males and 55 females, with age 52(37,66)years old.The bone marrow smear prepared with Wright-Giemsa, The AI analysis system and manual audit were applied to classify 13 types of bone marrow nucleated cell, taking the results of manual audit as the gold standard, comparing the difference between the results of the two methods, using statistical software to draw the confusion matrix, The compliance between the manual audit results and the pre-classification results of the AI analysis system was calculated by the Kappa consistency test method; The consistency analysis between the pre-classification results of AI and those of the manual microscopic examination was performed by the Pearson test; (2)Statistics analyzed the blast cell differential count differences of AI and manual microscopy, to evaluate the clinical application value of AI analysis system, which soured from thirty bone marrow samples of patients diagnosed with MDS and AML. Results:76 630 images of 13 nucleated cells were obtained by AI analysis system; the weighted average experimental diagnostic efficiency parameters of 13 types of bone marrow nucleated cells, are as follows: sensitivity(%)=95.82, specificity(%)=99.19, accuracy(%)=98.89, false positive rate(%)=0.81, false negative rate (%)=4.18; the correlation results, between the pre-classification results of AI and manual microscopic classification results,showed that blast cell, promyelocytes, neutrophilic myelocyte, neutrophilic metamyelocyte, band neutrophil, segmented neutrophi,eosinophil, basophil, polychromatic erythroblast, orthochromatic erythroblast, and lymphocytes have good positive correlation ( r>0.70,all P<0.001), while basophilic erythroblast and monocytes have no obvious correlation ( r=0.32,0.30, all P> 0.001); the count results of the blast cells in bone marrow smears of MDS and AML, got by AI and manual microscopy respectively, showed that the average percentage of blast cells was 8.19% by AI and 8.68% by manual microscopy in MDS, there was no significant difference between the two methods ( P>0.05); the average percentage of blast cells was 48.52% by AI analysis system and 53.77% by manual microscopy in AML, and although there was a significant difference in blast cell count ( P<0.01), coincidence the classification diagnostic criteria for AML (blast cells ≥ 20%). Conclusion:The AI analysis system performed good sensitivity, specificity and accuracy for 13 types of bone marrow nucleated cells, which showed potential application value for the rapid classification and diagnosis of MDS and AML.

Study on mechanism of Bupi Yichang pill in alleviating experimental ulcerative colitis by restoring the homeostasis of CD4+T cell subpopulations / 天津医药

Objective To investigate the regulatory effect of Bupi Yichang pill(BPYCP)on CD4+T cell subsets of ulcerative colitis(UC)mice.Methods Forty-eight C57BL/6 mice were randomly divided into 4 groups:the control group(n=10),the model group(DSS group,n=13),the model +BPYCP group(DSS+BPYCP group,n=13)and the model+ mesalazine(5-ASA)group(DSS+5-ASA group,n=12).The mouse UC model was induced by 2.5%dextrosan sulfate(DSS)solution.The DSS+BPYCP group and the DSS+5-ASA group were given BPYCP or 5-ASA for 2 weeks,respectively,and fecal viscosity and blood in stool were observed.The colon length was measured.Colonic mass index and unit colonic mass index were calculated.Hematoxylin-eosin(HE)staining was used to observe pathological changes of colon and to score the pathological tissue damage.The level of CD4+T cell subsets in mesenteric lymph nodes was detected by flow cytometry.The expression levels of cytokines interferon-γ(INF-γ),interleukin(IL-4),IL-17A,IL-10 and IL-21 secreted by CD4+T cell subsets in colon tissue were detected by ELISA.Real-time fluorescence quantitative PCR was used to detect colon tissue CD4+T cell subset nuclear transcription factors,mRNA expression levels of T-frame protein 21(T-bet),GatA-binding protein 3(GATA-3),retinoa-associated nuclear orphan receptor γt(RORγt),B cell lymphoma-6(Bcl-6)and Foxp3 in rats.Results Compared with the DSS group,the diarrhea and hematostoecium symptoms of UC mice in the DSS+BPYCP group and the DSS+5-ASA group were significantly improved,body weight and colon length of mice were increased,and colon mass,colon mass index and unit colon mass index were decreased(P＜0.05).The mucosal epithelium was more complete than that in the DSS group,and gland arrangement was more regular.The inflammatory cell infiltration was less,and the pathological tissue damage score was significantly decreased(P＜0.01).The proportion of Th2 cells in mesenteric lymph nodes was decreased,the proportion of Th17 cells and the level of IL-17A were decreased,and the mRNA levels of T-bet,GATA-3,RORγt and Bcl-6 in colon tissue were decreased(P＜0.05).In the DSS+BPYCP group,the proportion of Th1 cells decreased,the proportion of CD4+CD25+Treg cells,CD4+CD25+Foxp3+Treg cells and the level of IL-10 increased,and the proportion of CD4+CXCR5+Tfh cells and the level of IL-21 decreased.The level of Foxp3 mRNA increased(P＜0.05).The proportion of Th1 cells and the level of IFN-γ were decreased in the DSS+5-ASA group(P＜0.05).Conclusion BPYCP may alleviate UC by remodeling the homeostasis of CD4+T cell subpopulations.

Perilla frutescens PfFUL Regulates the Flowering and Silique Development in Transgenic Arabidopsis thaliana / 中国生物化学与分子生物学报

The FRUITFULL (FUL) gene belongs to the AP1/ FUL subfamily of the plant MADS-box family and has functions in regulating flowering time, floral meristem differentiation and fruit development. PfFUL gene sequence was derived from the perilla transcriptome data, and the basic physicochemical properties of PfFUL were analyzed by bioinformatics methods. Evolutionary relationships of PfFUL with other species of FUL were analyzed by phylogenetic tree. Plant expression vector 35S::PfFUL was constructed and used to transform wild type Col-0 and mutant ful-7 Arabidopsis to obtain overexpression 35S::PfFUL/ Col-0 and complemented mutation 35S::PfFUL / ful-7 plants respectively. Comparative phenotypic analysis was performed to preliminarily clarify the function of PfFUL gene in plant flowering and fruit development. The functions of the PfFUL gene during flowering and angular fruit development of the plants were initially clarified. The CDS of PfFUL gene is 738 bp and encodes 245 amino acids. Phylogenetic tree showed that the perilla PfFUL was closely related to Solanum lycopersicum, Salvia splendens and Salvia hispanica, but far related to Arabidopsis thaliana, Nicotiana tabacum and Vitis vinifera. Compared to Col-0 and ful-7, the transgenic plants showed early flowering (P0. 05), and the amount of wrinkled seed was significantly reduced (P<0. 01). In addition, phenotypic observations revealed that the transgenic plants also exhibited increased internode length and narrowed and curled cauline leaves. In conclusion, this study confirmed that the PfFUL gene regulates early flowering and fruit development in plants and participates in nutritional growth.

Association Between Plasma Homocysteine Level and Hyperuricemia in Elderly Patients With Hypertension / 中国医学科学院学报

Objective To explore the association between plasma homocysteine (Hcy) level and hyper-uricemia (HUA) in the elderly patients with hypertension.Methods From March to August in 2018,9902 hypertensive patients ≥ 60 years were routinely tested for blood biochemical indicators in Wuyuan county,Jiangxi province.The patients were assigned into a HUA group and a normal uric acid group.Multivariate Logistic regression was adopted to analyze the relationship between Hcy level and the risk of HUA.Results Compared with the normal uric acid group,the HUA group showed increased incidence of hyperhomocysteinemia (99.9% vs.98.7%,P<0.001) and elevated Hcy level[16.8 (13.8-21.5) μmol/L vs.14.4 (12.3-17.7) μmol/L,P<0.001].The multivariate Logistic regression analysis showed that after adjusting for influencing factors,the risk of HUA in the patients with hyperhomocysteinemia was 2.92 times of that in the patients with a normal Hcy level.The threshold effect analysis showed that the Hcy level was positively correlated with the occurrence of HUA in the case of Hcy<20 μmol/L (OR=1.05,95%CI=1.04-1.07,P<0.001).In the case of Hcy ≥ 20 μmol/L,there was no correlation between Hcy level and HUA (OR=1.00,95%CI=0.99-1.00,P=0.055),and the likelihood ratio test showed statistically significant results (P<0.001).Conclusion The elderly with hypertension should pay attention to control the Hcy level,which will be helpful to prevent the occurrence of HUA.

Analysis of biofilm-forming ability and drug resistance for Hypervirulent Klebsiella pneumoniae / 中华预防医学杂志

Investigate the biofilm-forming ability and drug resistance of Hypervirulent Klebsiella pneumoniae (HvKP) to provide scientific basis for the treatment of HvKP-infection. A total of 96 Klebsiella pneumoniae strains isolated from clinical infection specimens in Changsha Central Hospital from January to December in 2021 were retrospectively collected, and the clinical data of patients were collected. The string test preliminarily distinguished between HvKP and classic Klebsiella pneumoniae (CKP). The biofilm-forming ability of clinical strains of Klebsiella pneumoniae (KP) was determined by microplate method. The Vitek 2 Compact automatic microbial identification/drug sensitivity analysis system was used for bacterial identification and drug sensitivity test. The clinical data of patients, biofilm forming ability and drug resistance in the HvKP group and those in the CKP group were compared and analyzed. The results showed that a total of 20 strains of HvKP were isolated from 96 non-repetitive KP, and the detection rate was 20.8%. HvKP mainly come from respiratory specimens, up to 75.0%.The prevalence of hepatobiliary diseases and the infection rate of multiple sites in patients with HvKP infection were higher than those in patients with CKP infection, and the difference was statistically significant(χ2=5.184,7.488;P=0.023,0.006).There was no significant difference between the two groups in terms of gender, age, ICU admission, hypertension, diabetes, coronary heart disease, lung disease, urinary system disease, central nervous system disease and laboratory test indexes (all P>0.05).17 (85.0%) strains of HvKP can form biofilm, including 2 strains with weak biofilm-forming ability (10.0%), 10 strains with moderate biofilm-forming ability (50.0%) and 5 strains with strong biofilm-forming ability (25.0%). Among the 76 CKP, 71 (93.4%) could form biofilm, including 13 (17.1%) with weak biofilm-forming ability, 30(39.5%) with moderate biofilm-forming ability and 28 (36.8%) with strong biofilm-forming ability. There was no significant difference in biofilm-forming ability between HvKP and CKP (χ2=1.470,P=0.225).The overall resistance rate of HvKP was not high, but a multi-resistant HvKP resistant to carbapenems was found. The detection rate of multi-resistant HvKP (5.0%) was lower than that of multi-resistant CKP (28.9%), and the difference was statistically significant (χ2=4.984, P=0.026).The resistance rate of HvKP to piperacillin/tazobactam, aztreonam, ciprofloxacin, levofloxacin, ceftazidime, cefepime, tobramycin, minocycline, doxycycline, and compound sulfamethoxazole was lower than that of CKP, and the difference was statistically significant (all P<0.05). In conclusion, most of hypervirulent Klebsiella pneumoniae can form biofilm in this study, but the difference of biofilm-forming ability is not obvious compared with classic Klebsiella pneumoniae. HvKP maintains high sensitivity to commonly used antibacterial drugs, but the drug resistance monitoring of the bacteria cannot be ignored.

Analysis of biofilm-forming ability and drug resistance for Hypervirulent Klebsiella pneumoniae / 中华预防医学杂志

Investigate the biofilm-forming ability and drug resistance of Hypervirulent Klebsiella pneumoniae (HvKP) to provide scientific basis for the treatment of HvKP-infection. A total of 96 Klebsiella pneumoniae strains isolated from clinical infection specimens in Changsha Central Hospital from January to December in 2021 were retrospectively collected, and the clinical data of patients were collected. The string test preliminarily distinguished between HvKP and classic Klebsiella pneumoniae (CKP). The biofilm-forming ability of clinical strains of Klebsiella pneumoniae (KP) was determined by microplate method. The Vitek 2 Compact automatic microbial identification/drug sensitivity analysis system was used for bacterial identification and drug sensitivity test. The clinical data of patients, biofilm forming ability and drug resistance in the HvKP group and those in the CKP group were compared and analyzed. The results showed that a total of 20 strains of HvKP were isolated from 96 non-repetitive KP, and the detection rate was 20.8%. HvKP mainly come from respiratory specimens, up to 75.0%.The prevalence of hepatobiliary diseases and the infection rate of multiple sites in patients with HvKP infection were higher than those in patients with CKP infection, and the difference was statistically significant(χ2=5.184,7.488;P=0.023,0.006).There was no significant difference between the two groups in terms of gender, age, ICU admission, hypertension, diabetes, coronary heart disease, lung disease, urinary system disease, central nervous system disease and laboratory test indexes (all P>0.05).17 (85.0%) strains of HvKP can form biofilm, including 2 strains with weak biofilm-forming ability (10.0%), 10 strains with moderate biofilm-forming ability (50.0%) and 5 strains with strong biofilm-forming ability (25.0%). Among the 76 CKP, 71 (93.4%) could form biofilm, including 13 (17.1%) with weak biofilm-forming ability, 30(39.5%) with moderate biofilm-forming ability and 28 (36.8%) with strong biofilm-forming ability. There was no significant difference in biofilm-forming ability between HvKP and CKP (χ2=1.470,P=0.225).The overall resistance rate of HvKP was not high, but a multi-resistant HvKP resistant to carbapenems was found. The detection rate of multi-resistant HvKP (5.0%) was lower than that of multi-resistant CKP (28.9%), and the difference was statistically significant (χ2=4.984, P=0.026).The resistance rate of HvKP to piperacillin/tazobactam, aztreonam, ciprofloxacin, levofloxacin, ceftazidime, cefepime, tobramycin, minocycline, doxycycline, and compound sulfamethoxazole was lower than that of CKP, and the difference was statistically significant (all P<0.05). In conclusion, most of hypervirulent Klebsiella pneumoniae can form biofilm in this study, but the difference of biofilm-forming ability is not obvious compared with classic Klebsiella pneumoniae. HvKP maintains high sensitivity to commonly used antibacterial drugs, but the drug resistance monitoring of the bacteria cannot be ignored.

Antimicrobial resistance of bacteria:surveillance report from Hunan Pro-vincial Antimicrobial Resistance Surveillance System,2012-2021 / 中国感染控制杂志

Objective To understand the distribution of frequently isolated pathogenic bacteria from clinical speci-mens and their antimicrobial resistance changes in Hunan Province from 2012 to 2021,and to provide scientific evi-dence for the formulation and evaluation of antimicrobial clinical administration policies.Methods Species identifi-cation,selection of quality control strains and antimicrobial susceptibility testing agents were conducted according to the technical scheme of the China Antimicrobial Resistance Surveillance System(CARSS).Duplicate strains were excluded based on the principle of counting the first strain in each case.Statistical analysis was performed by WHO-NET 5.6 software.The the variations in constituent ratio and resistance rate of strains were analyzed with linear trend test,and the magnitude of change was described with Pearson correlation coefficient.Results From 2012 to 2021,the number of clinically isolated bacteria in the analysis increased from 82 759 to 312 914,with Gram-negative bacteria accounting for 69.5％-72.4％.The major Gram-positive bacteria were Staphylococcus aureus,Staphylo-coccus epidermidis,Streptococcus pneumoniae,Enterococcus faecalis and Enterococcus faecium,and the major Gram-negative bacteria were Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa,Acinetobacter baumannii and Enterobacter cloacae.Isolation rate of Gram-positive bacteria increased yearly(r=0.022,P=0.001).Isolation rate of methicillin-resistant Staphylococcus aureus(MRSA)decreased from 34.3％to 24.8％.Isolation rates of vancomycin-resistant Enterococcus faecium and Enterococcus faecalis were less than 3％and 2％,respectively,presenting a downward trend.The detection rate of penicillin-resistant Streptococcus pneumoniae(PRSP)decreased from 5.6％to 1.0％.Except cefoperazone sulbactam,resistance rates of Escherichia coli to other tested antimicrobial agents showed decreasing trends(r＜0,P=0.001).Isolation rates of third-generation cephalo-sporin-resistant Escherichia coli(CTX/CRO-R-EC)and carbapenem-resistant Escherichia coli(CREC)decreased year by year(from 70.5％to 45.3％,and 12.2％to 2.0％,respectively).Resistance rates of Klebsiella pneumo-niae to imipenem and meropenem have increased year by year,reaching 9.1％and 11.0％respectively in 2021,while isolation rate of carbapenem-resistant Pseudomonas aeruginosa(CRPA)decreased from 28.5％to 15.0％.Resistance rates of Acinetobacter baumannii to most antimicrobial agents were 40％-60％,and remained relatively stable.Isolation rate of carbapenem-resistant Acinetobacter baumannii(CRAB)ranged from 39.5％ to 59.6％.Conclusion The clinical isolation rates of most important special antimicrobial-resistant bacteria have been decrea-sing year by year,while the resistance rate of Klebsiella pneumoniae to carbapenem agents gradually increased.Antimicrobial stewardship as well as the prevention and control of healthcare-associated infection on specific antimi-crobial-resistant bacteria should continue to be implemented in the future.The coverage and quality of antimicrobial resistance surveillance in H unan Province should continue to be improved.

Antimicrobial resistance of bacteria from pleural and peritoneal effusion:surveillance report from Hunan Provincial Antimicrobial Resistance Sys-tem,2012-2021 / 中国感染控制杂志

Objective To understand the distribution and antimicrobial resistance changes of bacteria isolated from pleural and peritoneal effusion in Hunan Province,and provide reference for correct clinical diagnosis and rational antimicrobial use.Methods Data reported by member units of Hunan Provincial Antimicrobial Resistance Survei-llance System from 2012 to 2021 were collected.Bacteria antimicrobial resistance surveillance method was imple-mented according to technical scheme of China Antimicrobial Resistance Surveillance System(CARSS),and WHO-NET 5.6 software was used to analyze the data of bacteria isolated from pleural and peritoneal effusion as well as antimicrobial susceptibility testing results.Results From 2012 to 2021,a total of 28 934 bacterial strains were iso-lated from specimens of pleural and peritoneal effusions from member units of Hunan Provincial Antimicrobial Re-sistance Surveillance System,with 5 752 strains from pleural effusion and 23 182 from peritoneal effusion.The top five bacteria isolated from pleural effusion were Escherichia coli(n=907,15.8％),Staphylococcus aureus(n=535,9.3％),Klebsiella pneumoniae(n=369,6.4％),Staphylococcus epidermidis(n=452,7.9％),and Staphy-lococcus haemolyticus(n=285,5.0％).The detection rate of methicillin-resistant Staphylococcus aureus(MR-SA)from pleural effusion was 24.3％-39.2％,and that of methicillin-resistant coagulase negative Staphylococcus(MRCNS)was 58.8％-77.1％.The top five bacteria isolated from peritoneal effusion were Escherichia coli(n=8 264,35.6％),Klebsiella pneumoniae(n=2 074,9.0％),Enterococcus faecium(n=1 458,6.3％),Staphylo-coccus epidermidis(n=1 383,6.0％),and Pseudomonas aeruginosa(n=1 152,5.0％).The detection rate of MRSA from peritoneal effusion was 22.1％-52.4％,which presented a decreasing trend(P=0.004).The detec-tion rate of MRCNS was 60.4％-79.4％.The resistance rates of Enterobacterales from peritoneal effusion to ce-fazolin,cefuroxime,ceftriaxone and cefepime all showed decreasing trends(all P＜0.05).Vancomycin-,linezo-lid-,and teicoplanin-resistant Staphylococcus strains were not found in pleural and peritoneal effusions.The resis-tance rates of Enterococcus faecium to most tested antimicrobial agents were higher than those of Enterococcus fae-calis.The resistance rates of Enterobacterales to imipenem and meropenem were ≤8.5％.The resistance rates of non-fermentative Gram-negative bacilli to imipenem and meropenem were ≤43.3％.Conclusion The data structure of Hunan Antimicrobial Resistance Surveillance System for pleural and peritoneal effusions from 2012 to 2021 is relatively complete.The constituent and antimicrobial susceptibility of isolated pathogenic bacteria vary in different years.

Antimicrobial resistance of Pseudomonas aeruginosa:surveillance report from Hunan Provincial Antimicrobial Resistance Surveillance System,2012-2021 / 中国感染控制杂志

Objective To understand the distribution and changes in antimicrobial resistance of clinically isolated Pseudomonas aeruginosa(P.aeruginosa)in the member hospitals of Hunan Provincial Antimicrobial Resistance Surveillance System from 2012 to 2021.Methods Antimicrobial susceptibility testing by disk diffusion or automa-ted instrument was performed on clinical isolates.Testing results were determined according to the standards of 2022 edition from American Clinical Laboratory Standards Institute(CLSI).Statistical analysis was performed by WHONET 5.6 software.Data were analyzed by trend test(Cochran-armitage)and Chi-square test with SPSS.Results A total of 176 441 strains of P.aeruginosa were surveilled by Hunan Provincial Antimicrobial Resistance Surveillance System from 2012 to 2021.99.4％of the strains were isolated from hospitalized patients,and about 70％of the strains were isolated from respiratory specimens.8.4％of P.aeruginosa were from children(0-17 years old),91.6％were from adults.Antimicrobial susceptibility testing results showed that P.aeruginosa was most sensitive to polymyxin B over 10 years,with a resis-tance rate of less than 6％.Resistance rates to piperacil-lin,piperacillin/tazobactam,ceftazidime,cefepime,aztreonam,imipenem,amikacin,gentamicin,tobramycin,cip-rofloxacin,levofloxacin,and polymyxin B all showed downward trends.A total of 29 920 carbapenem-resistant P.aeruginosa(CRPA)strains were detected.The average isolation rate of CRPA in this province was 18.0％over 10 years.CRPA detection rate from adult was 18.5％,higher than that from children(12.3％),and both showing downward trends.Conclusion The resistance rate of clinically isolated P.aeruginosa in Hunan Province to most commonly used antimicrobial agents is decreasing.

Antimicrobial resistance of Acinetobacter baumannii:surveillance report from Hunan Provincial Antimicrobial Resistance System,2012-2021 / 中国感染控制杂志

Objective To understand the epidemiology of clinically isolated Acinetobacter baumannii(A.bauma-nnii)in Hunan Province.Methods Bacterial antimicrobial resistance surveillance was carried out according to the requirements of the technical program of National Antimicrobial Resistance Surveillance System.Clinical data of Acinetobacter spp.reported to Hunan Provincial Antimicrobial Resistance Surveillance System by multiple centers in Hunan Province from 2012 to 2021 were summarized and analyzed with reference to the standards of the American Clinical and Laboratory Standards Institute.Results A total of 169 438 strains of Acinetobacter spp.were detected during the 10-year period,with the detection rate of A.baumannii being the highest(82.74％).70 923 strains(53.63％)of carbapenem-resistant A.baumannii(CRAB)and 58 149 strains(43.97％)of carbapenem-sensitive A.baumannii(CSAB)were detected respectively.Both CRAB and CSAB were detected most frequently in the age group＞70 years,which were 34.44％and 32.02％,respectively.The percentage of CRAB and CSAB detected in the intensive care unit were 34.80％and 11.31％,respectively.CRAB and CSAB were mainly isolated from spu-tum/bronchoalveolar lavage fluid,followed by pus/secretion,urine,and blood.The resistance rates of CRAB to commonly used antimicrobial agents didn't change much during the 10-year period.Resistance rates of CRAB to ceftazidime and cefepime were both＞84％,to ampicillin/sulbactam and piperacillin/tazobactam were both＞82％,to aminoglycosides and quinolones were both＞59％,to minocycline and polymyxin B were 15.9％-25.0％and 1.3％-6.9％,respectively.CSAB were sensitive to commonly used antimicrobial agents.Conclusion The isolation rate of CRAB is high and there is no significant change in resistance to commonly used antimicrobial agents.