Culture of glioblastoma U87 stem-like cells and identification of its metabolic phenotype and tumorigenic ability / 基础医学与临床

Objective To cultivate glioblastoma U87 stem-like cells(SLCs)and to detect the level of stemness bio-markers,mitochondrial respiratory capacity and the capacity of in vivo tumorigenesis.Methods B-27,growth factors EGF and bFGF was added into DMEM/F-12 culture in serum-free stem cell culture medium for U87 SLCs.Suspended culture of U87 SLCs was suspended using the neuro-sphere formation assay,while adherent culture of U87 SLCs was achieved by coating Matrigel matrix on the culture surface.The mRNA and protein level of stemness biomarkers in culture were detected using real-time quantitative PCR and Western blot.The proportion of CD133+cells in culture was detected by flow cytometry.The changes of cell oxygen consumption rate were detected by Seahorse cell metabo-lism analysis.Cell tumorigenesis ability was verified by subcutaneous tumor transplantation in animals.Results U87 SLCs in stem cell culture medium would grow into typical sphere morphology within one week,and the spheres would continue to grow as the culture process prolongs.At the appropriate concentration of adhesive,U87 SLCs adhered to and grow well in stem cell culture medium.The mRNA transcription of stemness biomarkers such as CD133,nes-tin,OLIG2,CD44,CD15,and integrin α6(ITGA6)was significantly increased as found in both culture methods,and the protein levels of CD133 and nestin were also increased under both methods(P＜0.05).U87 SLCs showed higher mitochondrial reserve respiratory capacity(P＜0.05).U87 SLCs could form larger subcutaneous tumors with fewer inoculated cells(P＜0.05),and grew faster in vivo with stronger tumorigenic ability.Conclusions U87 SLCs have typical stemness characteristics and may function as tumor cell model with higher stemness properties.

Comparison of protocols for constructing animal models of early traumatic knee osteoarthritis / 中国组织工程研究

BACKGROUND:Current osteoarthritis modeling methods include anterior cruciate ligament transection(ACLT)and ACLT combined with medial meniscal anterior horn resection.ACLT requires excessive postoperative exercise,which is time and labor-intensive.Complete removal of anterior horn of the medial meniscus can cause collateral damage and increase variability in modeling outcomes,requiring higher surgical skills from the surgeon. OBJECTIVE:To modify and simplify the traditional method to create animal osteoarthritis model and compare osteoarthritis symptoms of different modeling methods under a low-load exercise environment. METHODS:Forty-eight Sprague-Dawley rats were randomly assigned in four groups(n=12 per group):sham operation(complete exposure of the knee cavity of the left hind limb followed by suturing the joint cavity and skin),ACLT,ACLT+anterior horn resection(removal of the anterior horn of the medial meniscus)and ACLT+anterior horn tear(anterior horn tear of the medial meniscus).At 4 weeks after modeling,the rats were euthanized and their knee specimens were collected for gross observation,X-ray and CT scans,pathological observation,and PCR detection. RESULTS AND CONCLUSION:Gross observation:Mild meniscal wear was observed in the ACLT group.In the ACLT+anterior horn tear group,severe wear of the lateral condyle articular surface,mild wear of the medial condyle articular surface,severe meniscal wear,and full wear of the medial meniscus were observed.The ACLT+resection group showed severe wear of the lateral condyle articular surface,mild wear of the medial condyle articular surface,absence of the anterior horn of the medial meniscus,and meniscus wear area＞50%.Imaging examinations showed no significant difference among the four groups.However,the anterior tibial translocation sign was observed in the three operation groups and the anterior horn of the medial meniscus was missing in the ACLT+anterior horn resection group.Histopathological section observation:Hematoxylin-eosin,toluidine blue,and Sirius red staining showed smooth joint surfaces in the sham operation group and ACLT group;cartilage damage and matrix degradation were evident in the ACLT+anterior horn tear and ACLT+anterior horn transection groups,with less cartilage damage and matrix degradation in the ACLT+anterior horn tear group.PCR results showed higher mRNA expressions of interleukin 1β,interleukin 6,interleukin 8,tumor necrosis factor α,matrix metalloproteinase 1 and matrix metalloproteinase 3 and lower mRNA expressions of aggrecan in the ACLT+anterior horn tear group and ACLT+anterior horn resection group than in the sham operation group and ACLT group(P＜0.05).The mRNA expressions of interleukin 6,matrix metalloproteinase 1,and matrix metalloproteinase 3 were higher in the ACLT + anterior horn resection group than in the ACLT +anterior horn tear group(P＜0.05).To conclude,ACLT alone is less likely to induce osteoarthritis with obvious cartilage wear.ACLT combined with anterior horn resection or tear of the medial meniscus can induce obvious symptoms of osteoarthritis and achieve similar modeling effects.

Evaluation and Consideration on Equity in Health Technology Assessment / 中国卫生经济

Health technology assessment(HTA)is an important tool to inform health decision-making.Although highly related to ethical issues in the context of HTA,equity has attracted much attention from the academia,a consensus has not yet been reached on how to define and evaluate equity in China and abroad.It introduces the concept of equity,pointes out the necessity to realize health equity and the reflection of equity in healthcare sector,and further elaborates four ways to consider equity,and described the official practice of equity in HTA at home and abroad.It proposes several suggestions for China's HTA:considering equity in HTA and the discussion of equity should depend on specific decision-making scenarios;clarifying what health measurement perspective should be adopted before measuring health equity;paying attention to the value judgment of equity adopted by various stakeholders;conducting basic researches on the general population's preference for health measurement perspectives and value judgments of equity in China in a gesture to improve the evaluation system of equity in HTA.

Effect of acupuncture treatment on nonketotic hyperglycemic hemichorea-hemiballismus: a case report.

Nonketotic hyperglycemic hemichorea-hemiballismus is a rare syndrome in the clinic, and treatment is often delayed. Hypoglycemic therapy is the most widely used and effective treatment, but some patients experience a slower improvement. Other symptomatic treatment medicines have some degree of side effects. Acupuncture treatment is beneficial for hemichorea-hemiballismus. A male patient, aged 59 years, first visited our hospital outpatient department due to motor agitation with involuntary movements of the right limb. He had a history of type 2 diabetes mellitus and poor blood glucose control. His serum glucose was 26.5 mmol/L (normal: 4.4-6.1 mmol/L), and magnetic resonance imaging demonstrated an irregular area of high signal intensity in T1-weighted imaging, low signal intensity on T2-weighted imaging, and high signal intensity in the left corpus striatum in T2-FLAIR imaging. Hospitalization was recommended for the patient. After ruling out other possibilities, he was eventually diagnosed with nonketotic hyperglycemic hemichorea-hemiballismus. Intensive glycemic control was immediately started with subcutaneous injection and acupuncture treatment at "governor vessel 13 acupoints", and the involuntary movements completely disappeared on the ninth day of hospitalization. The pathophysiology of nonketotic hyperglycemic hemichorea-hemiballismus is unclear. Different patient histories lead to different brain tissue conditions, and relapses and uncontrolled blood glucose add difficulties to treatment. According to Traditional Chinese Medicine theory, insufficient kidney essence leads to brain dystrophy and causes the symptoms of hemichorea-hemiballismus. Research evidence has shown that acupuncture at "governor vessel 13 acupoints" has a beneficial treatment effect on nonketotic hyperglycemic hemichorea-hemiballismus.

A double-blind, placebo-controlled, randomized withdrawal trial of sarilumab for the treatment of glucocorticoid-dependent sarcoidosis.

OBJECTIVES: Effective steroid-sparing therapies for the treatment of sarcoidosis are lacking; interleukin-6 (IL-6) antagonists may reduce sarcoidosis disease activity. This study assessed the safety and efficacy of the IL-6 receptor antagonist, sarilumab, in subjects with glucocorticoid-dependent sarcoidosis. METHODS: This phase II, double-blind, placebo-controlled, randomized withdrawal trial enrolled 15 subjects with biopsy-proven sarcoidosis at Stanford University from November 2019 to September 2022. In Period 1, subjects were treated with open-label sarilumab 200mg subcutaneously every two weeks for 16 weeks, with predefined tapering of prednisone. Subjects who completed Period 1 without a sarcoidosis flare entered Period 2 and were randomized to continue sarilumab or to receive matching placebo for 12 weeks. Endpoints included flare-free survival, as well as changes in pulmonary function tests, chest imaging, patient reported outcomes, and laboratory values. RESULTS: Fifteen subjects were enrolled in the study (median age 57 years, 80% male, 73.3% White), and 10 subjects successfully completed Period 1. During Period 1, 4 of 15 subjects (26.7%) discontinued due to worsening of their sarcoidosis, and CT chest imaging worsened in 5 of 15 subjects (35.7%). During Period 2, 0 of 2 subjects in the sarilumab group and 1 of 8 subjects (12.5%) in the placebo group had a flare. Treatment with sarilumab 200 mg was generally well tolerated in subjects with sarcoidosis. CONCLUSION: In this double-blind, placebo-controlled, randomized withdrawal trial, a meaningful signal for improvement in subjects with sarcoidosis treated with sarilumab was not observed. Given the small numbers in this study, no definitive conclusions can be drawn. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04008069.

Construction of a caries diagnosis model based on microbiome novelty score / 华西口腔医学杂志

OBJECTIVES@#This study aimed to analyze the bacteria in dental caries and establish an optimized dental-ca-ries diagnosis model based on 16S ribosomal RNA (rRNA) data of oral flora.@*METHODS@#We searched the public databa-ses of microbiomes including NCBI, MG-RAST, EMBL-EBI, and QIITA and collected data involved in the relevant research on human oral microbiomes worldwide. The samples in the caries dataset (1 703) were compared with healthy ones (20 540) by using the microbial search engine (MSE) to obtain the microbiome novelty score (MNS) and construct a caries diagnosis model based on this index. Nonparametric multivariate ANOVA was used to analyze and compare the impact of different host factors on the oral flora MNS, and the model was optimized by controlling related factors. Finally, the effect of the model was evaluated by receiver operating characteristic (ROC) curve analysis.@*RESULTS@#1) The oral microbiota distribution obviously differed among people with various oral-health statuses, and the species richness and species diversity index decreased. 2) ROC curve was used to evaluate the caries data set, and the area under ROC curve was AUC=0.67. 3) Among the five hosts' factors including caries status, country, age, decayed missing filled tooth (DMFT) indices, and sampling site displayed the strongest effect on MNS of samples (P=0.001). 4) The AUC of the model was 0.87, 0.74, 0.74, and 0.75 in high caries, medium caries, low caries samples in Chinese children, and mixed dental plaque samples after controlling host factors, respectively.@*CONCLUSIONS@#The model based on the analysis of 16S rRNA data of oral flora had good diagnostic efficiency.

Analysis of research hot-spots and development trends of county-level TCM hospitals in China based on CiteSpace / 国际中医中药杂志

Objective:To analyze the current development status, research hot-spots and development trends of county-level Chinese hospitals in China.Methods:By retrieving the papers of county-level Traditional Chinese Medicine (TCM) hospitals in China National Knowledge Infrastructure (CNKI), Chinese Science and Technology Periodicals Full-text Database (VIP), Wanfang and China Biomedical Database (SinoMed ) with CiteSpace 5.5.R2 software to draw maps of the author, institution and keyword and make analysis.Results:A total of 598 papers are included in this study, and the trend of publications shows an upward trend and then a downward trend. Researchers conduct research in the form of forming research teams. The research institutions are mainly medical colleges and scientific research institutions. The most frequently showed keyword is "medical reform". The keyword knowledge map shows that the research content mainly involves medical reform, problem analysis and countermeasures, economic operation efficiency, TCM service capability, data envelopment analysis and so on.Conclusions:The county-level TCM hospitals generally show sound development trend in China. The research trend is dominated by county-level TCM hospitals. Scale development has been transformed into the study of hospital operation efficiency. County-level TCM hospitals need to start from their own actual conditions, strengthen cooperation in the form of teams and strengthen academic cooperation among institutions, so as to realize the basic TCM services for ordinary people.

Function, structure and catalytic mechanism of sucrose phosphate synthase: a review / 生物工程学报

Sucrose is a natural product occurs widely in nature. In living organisms such as plants, sucrose phosphate synthase (SPS) is the key rate-limiting enzyme for sucrose synthesis. SPS catalyzes the synthesis of sucrose-6-phosphate, which is further hydrolyzed by sucrose phosphatase to form sucrose. Researches on SPS in recent decades have been focused on the determination of enzymatic activity of SPS, the identification of the inhibitors and activators of SPS, the covalent modification of SPS, the carbohydrate distribution in plants regulated by SPS, the mechanism for promoting plant growth by SPS, the sweetness of fruit controlled by SPS, and many others. A systematic review of these aspects as well as the crystal structure and catalytic mechanism of SPS are presented.

Clinical significance of mycoplasma pneumoniae-DNA load detection in alveolar lavage fluid of children with refractory mycoplasma pneumoniae pneumonia / 中国小儿急救医学

Objective:To investigate the clinical significance of mycoplasm pneumoniae-DNA(MP-DNA) load detected in bronchoalveolar lavage fluid of children with refractory mycoplasma pneumoniae pneumonia(RMPP) in the assessment of the condition.Methods:A total number of 98 hospitalized children with RMPP diagnosed in the Department of Pediatric Respiratory of Shengjing Hospital of China Medical University from May to November in 2018 were selected as the subjects.Bronchoscopy was performed on each subject and the alveolar lavage fluid was collected to detect the MP-DNA load quantitatively.According to the test results, all children were divided into three groups: high-load group(>10 6/ml, 28 cases), medium-load group(10 3 to 10 6/ml, 54 cases), and low-load group(≤10 3/ml, 16 cases). The clinical manifestations, relevant laboratory data, imaging data, and bronchoscopy findings of the children in different load groups were compared. Results:Compared with the low-load group and medium-load groups, the level of C-reactive protein was the highest in the high-load group with statistical significance( P<0.05). The difference of lactate dehydrogenase levels among the three groups was statistically significant( P=0.001). The positive incidences of nodular protuberances, mucosal erosion and necrosis, bronchial inflammatory stenosis and mucus plugs occlusion in the high-load group were 35.7%, 32.1% and 64.3%, respectively, which were significantly higher than those in the medium-load group(13.0%, 5.6%, 38.95) and low-load group(0, 0, 12.5%), and the differences were statistically significant( P<0.05). The imaging manifestation of large areas consolidation/lung atelectasis and pleural effusion in the high-load group were 100% and 60.7%, respectively, which were significantly higher than those in the medium-load group(61.1%, 27.8%)and low-load group(25.0%, 12.5%), and the differences were statistically significant( P<0.05). Compared with the low-load group and medium-load group, the hospitalization days was the longest in the high-load group, and the fever clearance time after the application of macroclides was also the longest( P<0.05). Conclusion:The MP-DNA load of children with RMPP is closely related with the clinical features.The clinical manifestations of children with higher DNA-load are more serious.The level of local MP-DNA load can guide clinicians to use drugs rationally, better predicting the outcome of the disease.

Application of the Nivolumab in the treatment of advanced non-small cell lung cancer / 国际肿瘤学杂志

Lung cancer has the highest morbidity and mortality among malignant tumors in our country.The treatment researches related to non-small cell lung cancer (NSCLC) such as targeted therapy and immunotherapy are the research hotspots in recent years.Nivolumab is the first programmed death-1 (PD-1)inhibitor which is allowed to enter clinical trials,and it is also the first approved drug for tumor immunotherapy in China.In June 2018,Nivolumab injection was marketed in China,and it was used in the second-line treatment for the patients with advanced NSCLC,opened up a new era of the lung cancer treatment in our country.Review the clinical research related to the Nivolumab and NSCLC,summarizing usage,dosage,adverse reactions,will provide a help for clinical application,bring more benefits for patients at the same time.

Research progress of MP-DNA load in Mycoplasma pneumoniae pneumonia / 国际儿科学杂志

Mycoplasma pneumoniae pneumonia is one of the most common community-acquired pneu-monia in children. Local MP-DNA load objectively reflects the balance of the immune clearance between the pathogen and host. In recent years,scholars have used different quantitative detection of MP-DNA to conduct a lot of research on different objects. It has been found that MP-DNA load can contribute to the early diagnosis of Mycoplasma infection and has association with the severity of the disease,it may guide the rational administra-tions for clinicians and can better predict the prognosis.

Value of ultrasonic calcifications in differential diagnosis of thyroid papillary carcinoma in Hashimoto's thyroiditis background / 中国基层医药

Objective To study the clinical value of ultrasonic calcifications in differential diagnosis of thyroid papillary carcinoma in Hashimoto''s thyroiditis background,to provide basis for clinical diagnosis and treatment.Methods The clinical data of 800 patientss with thyroid nodules who were examined by pathological examination were retrospectively analyzed.All patients underwent ultrasound examination,180 cases with Hashimoto''s thyroiditis were included in the study group,620 cases without Hashimoto''s thyroiditis were included in the control group.The gold standard was the result of pathology.The value of ultrasonography and calcifications for the identification of Hashimoto''s thyroiditis and thyroid papillary carcinoma was compared.Results The average diameter of the study group was (10.78±2.16)mm,which of the control group was (11.98±3.25)mm,there was no significant difference(t=2.153,P=0.083).The accurate rate of ultrasound in the differential diagnosis of benign thyroid nodules and papillary thyroid carcinoma in the study group was 76.67％(138/180),which in the control group was 80.81％(501/620),the difference was not statistically significant(χ2=0.898,P=0.098).There was no significant difference in coarse calcification rate and micro calcification rate of malignant nodules between the two groups(χ2=0.558,P=0.175).In the study group,the rate of micro calcification in malignant nodules was 12.07％,which was significantly higher than 53.13％ in benign nodules(χ2=32.142,P=0.001).In the control group,the micro calcification rate and coarse calcification rate of malignant nodules were 49.28％ and 12.08％,respectively,which were higher than 19.13％ and 6.05％ of benign nodules (χ2=67.368,7.056,P=0.001,0.009).The coarse calcification rate of benign nodules and malignant nodules in the study group were 6.25％ and 11.21％,respectively,there was no significant difference(χ2=1.098,P=0.062).Conclusion Whether associated with Hashimoto''s thyroiditis has no influence in the ultrasound differential diagnosis of thyroid papillary carcinoma,more micro calcification occurs in malignant nodules,but without the background of Hashimoto''s thyroiditis,coarse calcification can also be seen in malignant nodules.

The incidence of post-operative adhesion following transection of uterine septum: a cohort study comparing three different adjuvant therapies.

OBJECTIVE: To investigate the clinical efficacy of postoperative estrogen therapy, intrauterine device (IUD) and intrauterine balloon in preventing intrauterine adhesions after transcervical resection of septum (TCRS). STUDY DESIGN: 238 patients who underwent TCRS in our hospital from March 2012 to December 2013 were allocated into one of four groups. In Group 1 (50 patients), women received postoperative estrogen therapy. In Group 2 (59 patients), an intrauterine contraceptive device (IUD) was placed into the uterine cavity at the end of the operation. In Group 3 (75 patients), a Foley catheter with the balloon inflated with 4ml of normal saline solution was placed into the uterine cavity at the end of the operation for five days. In Group 4 (54 patients), women did not receive any of the above treatment (comparison group). All subjects underwent two further hysteroscopy, one and three months after the initial surgery. RESULTS: The intrauterine adhesion rates among the four groups at one month were 22.0%, 28.81, 26.7% and 24.1% (p>0.05); and at the third month were 0%, 1.7%, 1.3% and 3.4%, respectively (p>0.05). CONCLUSIONS: The use of postoperative estrogen therapy, IUD or intrauterine balloon did not appear to have any benefit in reducing the incidence of postoperative intrauterine adhesion formation.

Determination of Content and Related Substances of Troxipite Tablets by HPLC / 中国药师

Objective:To establish a method for the determination of content and related substances of troxipite tablets by HPLC. Methods:A Waters Symmetry-C18 (150 mm × 4. 6 mm,5 μm) column was used. The mobile phase was methanol -0. 4% phosphoric acid solution (50∶50). The flow rate was 1. 0 ml·min-1. The detection wavelength was 260nm. The column temperature was 35℃and the injection volume was 20 μl. Results:The linear range of troxipite was 3-75 μg·ml-1(r=0.999 7). The average recovery was 99. 7%,RSD=0. 95%(n=9). Conclusion:The method is simple, accurate and specific, and can be used in the quality control of troxipite tablets.

The expression of circulating microRNA-595 in patients with hepatitis B virus-related liver failure and its relationship with disease prognosis / 中华传染病杂志

Objective To measure the expression of circulating microRNA (miRNA)in patients with hepatitis B virus (HBV)-related liver failure and its relationship with disease prognosis.Methods The miRNA expressions in serum of 5 patients with HBV-related liver failure and 5 healthy control subjects were compared using Exiqon miRCURY LNATM miRNA microarray.The sera from 20 patients with chronic hepatitis B (CHB),20 patients hepatitis B related cirrhosis,50 patients with HBV-related liver failure and 40 healthy persons in Ruijin Hospital were collected.The relative expression of miRNA-595 was measured using quantitative real-time polymerase chain reaction (PCR).The relative expressions of miRNAs among groups were analyzed using student t test,the correlations were analyzed by Pearson and Spearman correlation.Results Microarray informed that 92 miRNAs changed significantly in patients with HBV-related liver failure,and miRNA-595 increased most significantly.The results of real-time PCR showed that the relative expressions of miRNA-595 ,miRNA-300 and miRNA-122 were 6.03 (t=3.134, P =0.003),3.12 (t=7.221 ,P <0.01)and 2.77 (t=2.671 ,P =0.021),which were higher compared to those in healthy control group.In the analysis of the relationship between miRNA-595 expression and disease prognosis in patients with HBV-related liver failure,the relative expressions of miRNA-595 in patients with CHB,hepatitis B related cirrhosis and HBV-related liver failure were 2.26 (t =3.780,P =0.001),3.32 (t = 6.111 ,P < 0.01)and 6.03 (t = 3.134,P = 0.003),respectively,which were all increased compared to that of the healthy control.The relative expression of miRNA-595 of patients with HBV-related liver failure was 2.66 times (t=2.450,P =0.043)higher than that of patients with CHB. When dividing patients according to prothrombin activity,miRNA-595 increased significantly in patients with early stage liver failure.When dividing patients according to model of end-stage liver disease (MELD) score,MELD score was positive correlated with the expression of miRNA-595 when MELD score was under 30 (r=0.673,P =0.004).The expression of serum miRNA-595 in survival group (11 .08,n=23) was higher than that in non-survival group (3.67,n = 27,t =4.309,P =0.041).Conclusions The expressions of miRNA595 ,miRNA-300 and miRNA-122 are all increased in patients with HBV-related liver failure,especially the expression of circulating miRNA-595 at early stage of the disease.The miRNA-595 may be used as a new serum biomarker for monitoring the severity of disease.