1.Action mechanism of Bushenhuoxue decoction on promoting nucleus pulposus-like differentiation of adipose-derived stem cells
Zehua GUO ; Zhaoyong LI ; Long CHEN ; Jiahao DUAN ; Haobo JIANG ; Guangxue CHEN ; Youxian SU ; Enxu LIU ; Shaofeng YANG
Chinese Journal of Tissue Engineering Research 2024;28(25):3974-3980
BACKGROUND:Stem cell transplantation is a new way to prevent and cure intervertebral disc degeneration.However,whether the transplanted stem cells can survive,proliferate,differentiate,and restore the function of nucleus pulposus cells after transplantation,is the key and difficult point to overcome. OBJECTIVE:To explore the effects of Bushenhuoxue decoction on survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells. METHODS:A Transwell chamber was used to construct a co-culture model of human adipose-derived stem cells and human degenerative nucleus pulposus cells.The experiment was divided into control group,model group,drug-containing serum group,and drug-free serum group.Except for the control group,the co-culture system of other groups was treated with 50 μmol/L tert-butyl hydrogen peroxide for 24 hours.The drug-containing serum group and drug-free serum group were treated with DMEM low-glucose complete culture medium containing drug-containing serum of Bushenhuoxue decoction or drug-free serum with 20%volume fraction for 48 hours.The sublayer adipose-derived stem cells were taken.Toluidine blue staining was used to detect proteoglycan synthesis levels.Real-time PCR method was used to detect mRNA expression of type Ⅱ collagen,proteoglycan and SRY-box transcription factor 9.The protein expression of SOX9 was detected by western blot assay.Lactate dehydrogenase assay was used to detect cytotoxicity.Flow cytometry was used to detect reactive oxygen species,and β-galactosidase staining was used to detect cell senescence. RESULTS AND CONCLUSION:(1)Compared with the control group,the proportion of necrotic cells in the model group increased;toluidine blue staining became lighter,and the expression levels of type Ⅱ collagen,proteoglycan,SOX9 mRNA and SOX9 protein decreased(P<0.05).Compared with the model group,the drug-containing serum of Bushenhuoxue decoction could significantly reduce cell injury and promote the expression of type Ⅱ collagen,proteoglycan,SOX9 mRNA,and SOX9 protein(P<0.05),but the improvement in the drug-free serum group was not significant(P>0.05).(2)Compared with the control group,the contents of cytotoxicity,reactive oxygen species,and cell senescence in the model group were significantly increased.Compared with the model group,the microenvironment of the coculture system was significantly improved by drug-containing serum of Bushenhuoxue decoction(P<0.05),while drug-free serum had no significant effect on the microenvironment of the co-culture system(P>0.05).(3)The results show that Bushenhuoxue decoction can promote the survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells.
2.Establishment and validation of a predictive model for the progression of pancreatic cystic lesions based on clinical and CT radiological features
Wenyi DENG ; Feiyang XIE ; Li MAO ; Xiuli LI ; Zhaoyong SUN ; Kai XU ; Liang ZHU ; Zhengyu JIN ; Xiao LI ; Huadan XUE
Chinese Journal of Pancreatology 2024;24(1):23-28
Objective:To construct a machine-learning model for predicting the progression of pancreatic cystic lesions (PCLs) based on clinical and CT features, and to evaluate its predictive performance in internal/external testing cohorts.Methods:Baseline clinical and radiological data of 200 PCLs in 177 patients undergoing abdominal thin slice enhanced CT examination at Peking Union Medical College Hospital from July 2014 to December 2022 were retrospectively collected. PCLs were divided into progressive and non-progressive groups according to whether the signs indicated for surgery by the guidelines of the European study group on PCLs were present during three-year follow-up. 200 PCLs were randomly divided into training (150 PCLs) and internal testing cohorts (50 PCLs) at the ratio of 1∶3. 15 PCLs in 14 patients at Jinling Affiliated Hospital of Medical School of Nanjing University from October 2011 to May 2020 were enrolled as external testing cohort. The clinical and CT radiological features were recorded. Multiple feature selection methods and machine-learning models were implemented and combined to identify the optimal machine-learning model based on the 10-fold cross-validation method. Receiver operating characteristics (ROC) curve was drawn and area under curve (AUC) was calculated. The model with the highest AUC was determined as the optimal model. The optimal model's predictive performance was evaluated on testing cohort by calculating AUC, sensitivity, specificity and accuracy. Permutation importance was used to assess the importance of optimal model features. Calibration curves of the optimal model were established to evaluate the model's clinical applicability by Hosmer-Lemeshow test.Results:In training and internal testing cohorts, the progressive and non-progressive groups were significantly different on history of pancreatitis, lesions size, main pancreatic duct diameter and dilation, thick cyst wall, presence of septation and thick septation (all P value <0.05) In internal testing cohort, the two groups were significantly different on gender, lesion calcification and pancreatic atrophy (all P value <0.05). In external testing cohort, the two groups were significantly different on lesions size and pancreatic duct dilation (both P<0.05). The support vector machine (SVM) model based on five features selected by F test (lesion size, thick cyst wall, history of pancreatitis, main pancreatic duct diameter and dilation) achieved the highest AUC of 0.899 during cross-validation. SVM model for predicting the progression of PCLs demonstrated an AUC of 0.909, sensitivity of 82.4%, specificity of 72.7%, and accuracy of 76.0% in the internal testing cohort, and 0.944, 100%, 77.8%, and 86.7% in the external testing cohort. Calibration curved showed that the predicted probability by the model was comparable to the real progression of PCLs. Hosmer-Lemeshow goodness-of-fit test affirmed the model's consistency with actual PCLs progression in testing cohorts. Conclusions:The SVM model based on clinical and CT features can help doctors predict the PCLs progression within three-year follow-up, thus achieving efficient patient management and rational allocation of medical resource.
3.Anti-pancreatic cancer effect of recombinant mouse peroxidase reductase-5 in vivo
Lin YANG ; Huiping XIE ; Miao WANG ; Jianing FENG ; Yuanyuan JIN ; Zhifei ZHANG ; Zhaoyong YANG
Chinese Journal of Immunology 2024;40(5):905-909
Objective:To investigate whether murine peroxidase reductase-5(mPRDX5)has anti-tumor activity in mice,so as to further confirm the anti-tumor activity and mechanism of recombinant peroxidase reductase-5.Methods:High purity mPRDX5 was obtained by heterologous expression and purification in vitro.Pancreatic cancer Pan02 cells were inoculated subcutaneously on the left axillary back of mice to establish a tumor bearing mouse model.Mice were randomly divided into PBS(solvent control)group,GEM(gemcitabine)50.0 mg/kg group and mPRDX5 10.0 mg/kg group,with 10 mice in each group,and the tumor related indexes were detected in mice.Results:Compared with PBS group,weight of tumor-bearing mice in GEM group was decreased obviously,while weight of mPRDX5 group was increased to a certain extent.Tumor growth was good in PBS group,according to tumor volume,com-pared with PBS group,tumor growth inhibition rates in D7 were 87.07%in GEM group and 52.82%in mPRDX5 10.0 mg/kg group,re-spectively;according to tumor weight,compared with PBS group,GEM group and mPRDX5 10.0 mg/kg group had tumor growth inhibi-tion rates of 95.39%and 48.33%in D7,respectively.Polarization state of macrophages in tumor tissues of mice in PBS group and mPRDX5 group was analyzed,and it was found that compared with PBS group,M1 macrophages expressing CD86 in tumor tissues of mice in mPRDX5 group were significantly increased,while M2 macrophages expressing CD206 were significantly decreased.Conclu-sion:mPRDX5 has significant anti-pancreatic cancer activity in mice,and the activity is exerted by promoting M1-type polarization of macrophages in the tumor microenvironment.
4.Effects of butylphthalide combined with hyperbaric oxygen therapy on post-stroke cognitive impairment
Zhi WANG ; Liangbing ZHAO ; Chao SUN ; Zhaoyong YU ; Hao WANG ; Ying ZHANG ; Guofang CHEN
Chinese Journal of Primary Medicine and Pharmacy 2023;30(4):497-500
Objective:To investigate the clinical efficacy of butylphthalide combined with hyperbaric oxygen therapy on post-stroke cognitive impairment in patients with acute ischemic stroke.Methods:A total of 90 patients with post-stroke cognitive impairment who were hospitalized within 72 hours of onset in Suining County People's Hospital from December 2019 to November 2020 were included in this study. They were randomly divided into a control group and an observation group ( n = 45/group). The control group was given conventional treatment and the observation group was given butylphthalide combined with hyperbaric oxygen therapy in addition to conventional treatment. The National Institutes of Health Stroke Scale score, Montreal Cognitive Assessment score, and Activities of Daily Living score were compared between the two groups before and after treatment. Results:Before treatment, there were no significant differences in the National Institutes of Health Stroke Scale score, Montreal Cognitive Assessment score, and Activities of Daily Living score between the two groups (all P > 0.05). At 14 days and 1 month after surgery, the National Institutes of Health Stroke Scale scores in the observation group were (4.02 ± 2.18) points and (3.21 ± 2.03) points, which were significantly lower than (5.21 ± 2.24) points and (4.62 ± 2.68) points in the control group ( t =2.55, 2.81, both P < 0.05). At 1 and 3 months after treatment, the Montreal Cognitive Assessment score in the observation group were (19.79 ± 5.67) points and (23.69 ± 2.67) points, which were significantly higher than (16.88 ± 5.12) points and (19.74 ± 2.29) points in the control group ( t = 2.56, 7.53, both P < 0.05). At 1 and 3 months after treatment, Activities of Daily Living scores in the observation group were (54.85 ± 5.69) points and (74.38 ± 4.98) points, which were significantly higher than (46.78 ± 6.24) points and (63.21 ± 5.24) points in the control group ( t = 6.41, 9.76, both P < 0.05). Conclusion:Butylphthalide combined with hyperbaric oxygen therapy for the treatment of post-stroke cognitive impairment in patients with acute ischemic stroke can alleviate neurologic deficits, and improve cognitive function and the ability of daily life.
5.Sports and health integration: logic, dilemma and alleviation
Yanfeng ZHANG ; Yibo GAO ; Sen LI ; Zhaoyong WEN ; Aoyu ZHANG ; Jin HE
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(6):481-487
By combing out the historical development logic of sports and health integration and the current development achievement, the underlying logic of sports and health integration is described systematically in multiple dimensions, and the prominent dilemmas in development are analyzed as follows: lack of residents' health literacy, excessive dependence on medical treatment, low bridging of talent team construction, lack of the guidance of funds centralization, insufficient financial support, imperfect management system and mechanism, and unspecific departmental collaboration.The strategies for solving the problems are as follows: strengthening the integration of sports science and clinical medicine, consolidating the technical integration of sports prescription and clinical standardization process, further improving the business integration of exercise intervention and clinical treatment, and enhancing the industrial integration of sports products and medical treatment.
6.Clinical evaluation of laboratory diagnosis of Mycoplasma pneumoniae
Liangyu WANG ; Xiaohua HAN ; Ran WEI ; Lina HAN ; Xijie LIU ; Jingyi LI ; Haiwei DOU ; Zhaoyong WU ; Shaogang LI ; Deli XIN ; Kunling SHEN
Chinese Journal of Applied Clinical Pediatrics 2022;37(16):1252-1256
Objective:To compare the laboratory diagnostic methods of Mycoplasma pneumonia(MP) and evaluate its clinical value.Methods:A prospective study.Throat swabs and double sera of children with MP infection were collected from December 2016 to January 2017 in Shengjing Hospital Affiliated to China Medical University; throat swab samples of healthy children aged 3 to 5 in Chaoyang District, Beijing were collected from March to May 2017.Passive agglutination (PA) was used to detect the double serum.Taking the 4-fold increase or decrease of the specific antibody titer of the double serum as the gold standard, the receiver operating characteristic curve (ROC) was drawn, and the laboratory methods for detecting MP infection were compared and evaluated.Results:(1)A total of 93 children with MP infection were clinically diagnosed, including 42 males (45.2%) and 51 females (54.8%), with an average age of 5.5 years.Sixty cases (64.5%) of MP infection were diagnosed.There were 349 healthy children, 198 males and 151 females, with an average age of 4.3 years.The positive rate of throat swab culture was 0.6% (2 cases), and the positive rate of fluorescent quantitative PCR(qPCR) was 18.9% (66 cases). (2) The culture specificity was the highest (100.0%) and the sensitivity was the lowest (65.0%). PA and enzyme linked immunosorbent assay (ELISA) were used to detect a single serum in the acute phase, the sensitivity was 71.7% and 86.5% respectively.ROC curve suggested that the current clinical diagnostic threshold MP specific antibody IgM ≥ 1∶160 was not the best diagnostic threshold.Molecular biological diagnostic methods were the most sensitive, RNA simultaneous and testing (SAT) was 85.0% and qPCR was 93.0%; while the specificity was low, 75.7% (SAT) and 63.6% (qPCR), respectively.(3) At the same time, MP nucleic acid (SAT, PCR) of throat swabs and a single serum (ELISA, PA) of children in acute phase were detected, the sensitivity was increased to 95.0%-100.0%, and the specificity was 63.6%-75.7%.Conclusions:Molecular biology is highly sensitive in diagnosing MP infection.It has asymptomatic infection or is carried after infection.Whether it needs treatment needs to be combined with clinical practice, when MP detection is positive.The detection of a single serum in the acute phase with a course of about 1 week has high sensitivity and is of reference value for the diagnosis of MP infection, but the diagnosis needs to be combined with clinical practice.The sensitivity and accuracy of detecting MP infection by single serological test combined with SAT in acute phase are higher than that by single application.
7.Epidemic and clinical characteristics of 1 976 children with hand-feet-mouth disease in Chengdu, 2018 to 2019
Wenxiao PANG ; Zhaoyong ZHANG ; Ying LAN ; Yilan ZENG
Chinese Journal of Infectious Diseases 2020;38(9):575-577
Objective:To analyze the epidemiological and clinical characteristics of hand-feet-mouth disease (HFMD).Methods:The clinical and pathogenic data of 1 976 cases with HFMD hospitalized in Public Health Clinical Center of Chengdu from August 2018 to September 2019 were retrospective analyzed.Results:Among the 1 976 cases of HFMD, 1 094 cases (55.36%) were one to two years old, and 803 cases (40.64%) lived in the main urban area. The majority were scattered patients (1 344 cases, 68.02%). There were 1 348(72.7%) of 1 854 cases mainly infected by Coxsackie virus 6 (CV-A6). The main distribution of rashes in children infected by CV-A6 were palm/soles, trunks, limbs, oral ulcer and perioral areal, lips and hip. The types of rash were maculopapule, vesicle, and bulla. Fever (1 543 cases, 78.09%) was the main concomitant symptom. Acute tonsillitis (811 cases, 41.04%), myocardial injury (767 cases, 38.82%), and herpetic angina (658 cases, 33.30%) were the most common complications. The incidence of onychomadesis were nine (7.03%) among the 128 patients during follow-up.Conclusions:CV-A6 is the main pathogen of the HFMD prevalence. Children less than two years old are susceptible and the main symptoms of HFMD are rash and fever.
8.A case-control study to identify risk factors associated with carbapenem resistant Enterobacteriaceae nosocomial infection
Zhaoyong ZHANG ; Hongwei YANG ; Fei YU ; Youqing WANG ; Fei ZHAO ; Jicai ZHANG
International Journal of Laboratory Medicine 2018;39(13):1573-1577,1581
Objective To investigate the characteristics of nosocomial infection of carbapenem resistant En-terobacteriaceae ,then to study the risk factors and prognosis of patients ,as to provide evidence for effective control of CRE hospital infection .Methods A retrospective epidemiology study was conducted of CRE infec-ted patients from January 2016 to June 2017 .This was a 1∶2 matched case-control study of patients with in-fection due to carbapenem resistant Enterobacteriaceae and randomly select 218 patients with non CRE infec-tion in the same hospital as control group ,then the risk factors of CRE infection were investigate .Results A-mong the 109 cases CRE infection patients ,the most common bacteria were Klebsiella pneumoniae ,Enter-obacter cloacae and Escherichia coli .The CRE strains were more sensitive to minocycline than other common antibiotics ,the resistance rate to other antibiotics was more than 40% .Univariate analysis showed that ICU staying time more than 7 d ,using beta lactamase inhibitors and carbapenem antibiotics ,combination therapy and mechanical ventilation were the potential risk factors of CRE nosocomial infection .The non conditional multivariate Logistic regression analysis showed that Check in ICU more than 7 d (OR= 4 .981 ,95% CI 2 .451-10 .122 ) ,the use of containing beta lactamase inhibitor antibiotics (OR= 3 .718 ,95% CI 2 .162-6 .394) ,use of carbapenem antibiotics (OR=3 .232 ,95% CI 1 .879-5 .561) ,and mechanical ventilation (OR=5 .26 ,95% CI 2 .576-10 .742) were independent risk factors of CRE nosocomial infection .The nosocomial in- fection CRE strain was with highly antibiotic resistance rate ,and the average hospitalization time and mortality were significantly higher than those of the control group .Conclusion The carbapenem resistant Enterobacte-riaceae infection had multiple independent risk factors ,strengthening of these independent risk factor control can effectively prevent the spread of CRE isolates infection .
9.The antibacterial effect of Fusidic acid on Mycoplasma pneumoniae and antibiotic resistant Mycoplasma pneumoniae in vitro
Ran WEI ; Dawei SHI ; Haiwei DOU ; Shaogang LI ; Zhaoyong WU ; Dongxing GUO ; Liangyu WANG ; Deli XIN
Chinese Journal of Applied Clinical Pediatrics 2018;33(10):787-790
Objective To investigate the antibacterial effect of Fusidic acid on Mycoplasma pneumoniae and antibiotic resistant Mycoplasma pneumoniae in vitro.Methods Twenty-eight clinical strains of Mycoplasma pneumoniae isolated from patients with respiratory tract infection at Beijing Friendship Hospital Affiliated to the Capital University of Medical Sciences from January to December 2016 and 2 Mycoplasma pneumoniae reference strains were enrolled.The minimum inhibitory concentration (MIC) of Fusidic acid and Azithromycin were determined by using micro-dilution ration method.The chessboard method was used to check the antibacterial effect of combination between Fusidic acid and Azithromycin.The antibacterial activity of the Fusidic acid was evaluated by measuring the antibacterial rate of different concentrations.Results One isolate showed no mutation in 23SrRNA,26 isolates had one point mutation in loci 2063 and 1 isolate had one point mutation in loci 2064 among the 28 clinical isolates.The findings by micro-dilution method results showed that the MIC values of all the clinical isolates with mutations associated with macrolide resistance to Azithromycin were > 1.000 0 mg/L,and the MIC values of all the clinical isolates with no mutations to azithromycin were < 0.500 0 mg/L.The findings by micro-dilution method results showed that the MIC value of Fusidic acid for Mycoplasma pneumonia and drug resistance Mycoplasma pneumoniae was 1.000 0 mg/L.The Fractional Inhibitory Concentration index of Fusidic acid and Azithromycin combination was ≤0.500 0 mg/L.When the concentration of the Fusidic acid was lower than or equal to 32 MIC,the antibacterial effect of Fusidic acid against Mycoplasma pneumoniae increased with its higher concentration.When the concentration of the Fusidic acid was lower than or equal to 8 MIC,the longer the strain was exposed to the drug,the stronger antibacterial effect was against Mycoplasma pneumoniae.Conclusion If the treatment of Mycoplasma pneumoniae infection is not effective or the infection of patient is combined with bacteria,the application or combination of Fusidic acid may inhibit pathogenic bacteria effectively.Of course,how to use Fusidic acid in clinical treatment needs further study and discussion.
10.Effect of preoperative respiratory exercise on pulmonary function and complications in elderly patients after abdominal operation
Qingfeng WANG ; Xianwen YUAN ; Zhaoyong DING
Chinese Journal of Primary Medicine and Pharmacy 2018;25(2):172-175
Objective To compare and analyze the effects of preoperative respiratory function exercise on postoperative pulmonary function and complications in elderly patients who undergoing abdominal surgery . Methods A total of 92 elderly patients who underwent abdominal surgery were recruited .They were randomly divided into observation group(46 cases) and control group(46 cases) according to the random number table method . The control group was treated with routine preoperative care methods , the observation group was given preoperative respiratory exercise on the basis of the control group .The pulmonary function tests were performed pre and post surgery.Then,the postoperative pulmonary complications ,VC,FEV1,FVC,MVV and the average length of hospital stay of the two groups were recorded and compared .Results After operation,the levels of VC,FEV1,FVC and MVV in the control group were (2.48 ±0.71)L,(51.40 ±10.10)L,(1.79 ±0.17)L,(62.40 ±13.00)L/min,respectively, which in the observation group were (2.89 ±0.72) L,(63.20 ±9.60) L,(2.21 ±0.16) L and(73.30 ± 12.70)L/min,respectively,the differences between the two groups were significant (t =2.750,5.743,12.202, 4.068,all P<0.05).The average hospitalization time in the observation group was (11.62 ±1.41) d,which was significantly shorter than (15.56 ±6.23) d in the control group (t =4.163,P <0.001).The incidence rate of pulmonary infection in the control group was 26.09%,which was significantly higher than 8.69% in the observation group(χ2 =4.842,P<0.05).Conclusion The respiratory function training can effectively reduce the incidence of postoperative pulmonary complications in elderly patients with abdominal surgery ,improve the patients ' postoperative pulmonary function, and shorten the patients ' hospital recovery time .Respiratory function exercise is a simple method,and is worthy of promotion in related fields .

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