Investigator-initiated clinical trials （IIT） are an important part of scientific and technological activities involving human study participants. Among them， high-quality IIT can be used to support the marketing and registration application of drugs， medical devices， and other products when conditions permit. Currently， there is a huge gap between IIT and industry-initiated clinical trials. The use of unlisted products in IIT has problems， such as lack of regulatory support， insufficient research funding support， the need to improve the ability of clinical research management departments， the weakness of professional clinical research teams， and the difficulty of ethics review to match the demands. The challenges could be addressed by improving regulations and conducting pilot trials on a small scale， guaranteeing adequate research funding， strengthening the construction of clinical research management systems， building professional clinical research teams， ensuring the quality of ethical reviews and strict follow-up reviews， shifting from ethical reviews to a system for protecting research participants， and reinforcing training for researchers. Ethics committees should strictly review key points， such as the risk-benefit ratio， informed consent， research funding， compensation for damages， qualifications and equipment of research team members， and management of conflict of interest.

To promote the development of extracellular vesicles of herbal medicine especially the establishment of standardization, led by the National Expert Committee on Research and Application of Chinese Herbal Vesicles, research experts in the field of herbal medicine and extracellular vesicles were invited nationwide with the support of the Expert Committee on Research and Application of Chinese Herbal Vesicles, Professional Committee on Extracellular Vesicle Research and Application, Chinese Society of Research Hospitals and the Guangdong Engineering Research Center of Chinese Herbal Vesicles. Based on the collation of relevant literature, we have adopted the Delphi method, the consensus meeting method combined with the nominal group method to form a discussion draft of "Consensus statement on research and application of Chinese herbal medicine derived extracellular vesicles-like particles (2023)". The first draft was discussed in online and offline meetings on October 12, 14, November 2, 2022 and April and May 2023 on the current status of research, nomenclature, isolation methods, quality standards and research applications of extracellular vesicles of Chinese herbal medicines, and 13 consensus opinions were finally formed. At the Third Academic Conference on Research and Application of Chinese Herbal Vesicles, held on May 26, 2023, Kewei Zhao, convenor of the consensus, presented and read the consensus to the experts of the Expert Committee on Research and Application of Chinese Herbal Vesicles. The consensus highlights the characteristics and advantages of Chinese medicine, inherits the essence, and keeps the righteousness and innovation, aiming to provide a reference for colleagues engaged in research and application of Chinese herbal vesicles at home and abroad, decode the mystery behind Chinese herbal vesicles together, establish a safe, effective and controllable accurate Chinese herbal vesicle prevention and treatment system, and build a bridge for Chinese medicine to the world.

In the implementation of exemption from ethical review,medical institutions equate exemption from ethical review with no ethical review or simple review,misunderstand the scope of exemption from ethical review,confuse the concepts of de-identification and anonymization,and equate privacy with personal information.The implementation faced challenges such as the coordination of conditions for exempt review with other regulations,the lack of clear decision-making subjects for exempting from ethical review,the legality and compliance of using general informed consent for biological samples and information data,as well as non-traceability and the risk of being re-identified of anonymous information for exemption from ethical review.Measures such as improving relevant laws and regulations,perfecting the construction and management of information databases and biological sample libraries,strengthening the project management and process supervision of exemption from ethical review,and implementing scientific review can ensure the legal and compliant implementation of exemption from ethical review by medical and health institutions.

Background: As Actinobacillus pleuropneumonniae (APP) infection causes considerable losses in the pig industry, there is a growing need to develop effective therapeutic interventions that leverage host immune defense mechanisms to combat these pathogens. Objectives: To demonstrate the role of microRNA (miR)-127 in controlling bacterial infection against APP. Moreover, to investigate a signaling pathway in macrophages that controls the production of anti-microbial peptides. Methods: Firstly, we evaluated the effect of miR-127 on APP-infected pigs by cell count/ enzyme-linked immunosorbent assay (ELISA). Then the impact of miR-127 on immune cells was detected. The cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 were evaluated by ELISA. The expression of cytokines (anti-microbial peptides [AMPs]) was assessed using quantitative polymerase chain reaction. The expression level of IL-6, TNF-α and p-P65 were analyzed by western blot. The expression of p65 in the immune cells was investigated by immunofluorescence. Results: miR-127 showed a protective effect on APP-infected macrophage. Moreover, the protective effect might depend on its regulation of macrophage bactericidal activity and the generation of IL-22, IL-17 and AMPs by targeting sphingosine-1-phosphate receptor3 (SIPR3), the element involved in the Toll-like receptor (TLR) cascades. Conclusions Together, we identify that miR-127 is a regulator of S1PR3 and then regulates TLRuclear factor-κB signaling in macrophages with anti-bacterial acticity, and it might be a potential target for treating inflammatory diseases caused by APP.

Objective:To explore the treatments and outcomes of heart and kidney transplantation(HKTx)and summarize its management experiences.Methods:From October 2016 to October 2020, clinical data, treatment strategies and prognosis of 11 patients received HKTx were analyzed retrospectively.In 11HKTx cases, the ratio of male-to-female was 10∶1, the age(50.6±12.9)years and the preoperative body mass index(26.72±3.29)kg/m 2.The preoperative cardiac function was class Ⅳ and the preoperative left ventricular ejection fraction(29.40±4.48)%.All patients were in uremic state pre-operation and underwent regular dialysis.The mean duration of dialysis was 2.5(0.5-7.0)years, preoperative creatinine 753.5(434-1144)μmol/L and preoperative predictive glomerular filtration rate 5.59(3.93-17.23)ml/(min preop 2). Non-staged transplant was performed and donor heart and kidney were from the same donor.The median time of cold cardiac ischemia 2.75(2.5, 4.0)hours, the median time of cold renal ischemia 9(8.5, 15.0)hours and the median time from the end of heart transplantation to the beginning of kidney transplantation 2(1.0, 3.5)hours.The immunosuppressive regimen was a combination of tacrolimus, mycophenolate mofetil and methylprednisolone. Results:Normal cardiac function and renal function normalized in 9 cases.At Month 6 post-operation, the postoperative left ventricular ejection fraction was(57.55±2.51)%, creatinine 107.7(85-132)μmol/L and urine volume in 24h 1988(1800-2200)ml.The long-term survival time was 6-62 months.No such complications as infection or rejection occurred in 9 patients.The cardiac function was class Ⅰ at Month 6 post-operation.One patient died from pulmonary mucor infection at Month 4 post-operation.Another death was due to gastrointestinal fungal infection at Month 1 after HKTx.Conclusions:HKTx is an effective treatment for end-stage heart disease with renal failure.

BACKGROUND: Accurate serum total thyroxine (TT4) measurement is important for thyroid disorder diagnosis and management. We compared the performance of six automated immunoassays with that of isotope-diluted liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) as the reference method. We also evaluated the correlation of thyroid stimulating hormone (TSH) with TT4 measured by ID-LC-MS/MS and immunoassays. METHODS: Serum was collected from 156 patients between October 2015 and January 2016. TT4 was measured by immunoassays from Abbott (Architect), Siemens (ADVIA Centaur XP), Roche (E601), Beckman-Coulter (Dxi800), Autobio (Autolumo A2000), and Mindray (CL-1000i), and by ID-LC-MS/MS. Results were analyzed using Passing-Bablok regression and Bland-Altman plots. Minimum requirements based on biological variation were as follows: a mean bias of ≤4.5% and total imprecision (CV) of ≤3.7%. RESULTS: All immunoassays showed a correlation >0.945 with ID-LC-MS/MS; however, the slope of the Passing-Bablok regression line varied from 0.886 (Mindray) to 1.23 (Siemens) and the intercept from −12.8 (Siemens) to 4.61 (Mindray). Only Autobio, Beckman-Coulter, and Roche included the value of one in the 95% confidence interval for slope. The mean bias ranged from −10.8% (Abbott) to 9.0% (Siemens), with the lowest value noted for Roche (3.5%) and the highest for Abbott (−10.8%). Only Abbott and Roche showed within-run and total CV ≤3.7%. CONCLUSIONS: Though all immunoassays correlated strongly with ID-LC-MS/MS, most did not meet the minimum clinical requirement. Laboratories and immunoassay manufacturers must be aware of these limitations.
Bias (Epidemiology) ; Diagnosis ; Humans ; Immunoassay ; Mass Spectrometry ; Methods ; Thyroid Gland ; Thyrotropin ; Thyroxine

We developed a method to identify serological humoral immunodominant proteinic antigen of Mycoplasma hyopneumoniae (Mhp). After constructing the recombinant plasmid pGEX-6P-1-mhp366 and transforming it into Escherichia coli BL21(DE3), the recombinant GST-Mhp366 protein was expressed successfully. The lysates of the recombinant GST-Mhp366 and genetic engineering GST were added into glutathione coated plates and reacted with 17 positive sera or 13 negative sera. Meanwhile, the optimization of experimental conditions, including coated antigen, blocking buffer, dilutions of sera and second antibody were determined. The optimal concentration of the coated antigen was the original bacteria lysates without dilution, and the optimal blocking buffer contained 10% FBS and 2.5% skim milk in PBS. Besides, the working concentration of serum samples and the HRP-tagged rabbit anti-pig IgG secondary antibody were 1:500 and 1:40 000, respectively. Thus, an indirect ELISA was established for identification of immunodominant protein antigens of Mhp. Meanwhile, this method was confirmed by the identified serological humoral immunodominant proteinic antigen Mhp156 and Mhp364. This method can be used for identification of the candidate vaccine antigens on a genome-wide scale. Furthermore, it can lay the foundation for identifying the candidate vaccine antigens through colostra and the nasal mucosal secretions.

Objective To summarize the outcomes and clinical experience of combined heart and kidney transplantation.Methods The clinical data of one case of combined heart and kidney transplantation were retrospectively analyzed.The kidney transplant was completed immediately after the heart transplant.The immunosuppressive therapy strategies included tacrolimus,corticosteroids and mycophenolate mofetil.Results For heart transplantation,heart cold ischemia time was 200 min,aorta blocking time was 136 min,and extracorporeal circulation time was 201 min.The kidney was transplanted to the right iliac fossa after heart transplantation.The endotracheal tube was removed 15 h after surgery.The patient was transferred to the general ward on the 8th day after surgery.The patient was discharged from the hospital at 27th day after surgery,the renal function was normal and no activity was restricted.Conclusion Reasonable perioperative management and selection of surgical methods are the keys to the success of combined heart and kidney transplantation.

OBJECTIVE:To establish HPLC fingerprints of Potentilla discolor,and to conduct authenticity identification.METHODS:HPLC method was adopted.The determination was performed on InertSustain C18 column with mobile phase consisted of 0.1％ formic acid solution-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min,detection wavelength of 360 nm,colunn temperature of 30 ℃,sample size of 10 μL.Using rutin as reference,HPLC chromatograms of 19 batches of P.discolor and 2 batches of P.chinesis were determined.TCM Fingerprint Similarity Evaluation System (2004) was used for similarity evaluation of 21 batches of samples,and common peak identification of 19 batches of P discolor SPSS 21.0 statisticl software was used for main component analysis and cluster analysis.RESULTS:There were 18 common peaks in HPLC fingerprints of 19 batches of P.discolor,the similarity was higher than 0.9.-HPLC chromatogram was in good agreement with control fingerprint.The similarity of 2 batches of P chinesis was lower than 0.7.The 21 batches of medicinal materials could be grouped into 2 categories,2 batches of P chinesis could be grouped into a category,19 batches of P.discolor could be grouped into a category.P discolor could be grouped into 4 categories.Rutin and quercitrin were main ingredients in 19 batches of P discolor.CONCLUSIONS:Established fingerprint can provide reference for authenticity identification and quality evaluation of P.discolor.

Objective To deveplope construct and validate a novel multiplex PCR system comprised of 30 Y-STR markers only with low and moderate mutation rates. Methods 30 Y-STRs characterized by low/moderate mutation rate and middle/high polymorphic was amplified simultaneously in a multiplex PCR system using the six color labeling fluorescence. PCR product was analyzed in a ABI 3500XL Genetic Analyzer. The accuracy, specifity, sensitivity and stability of the system and its validation on the mixtures were evaluated. Results The validation studies demonstrated that the system is a stable, accurate, and sensitive multiplex PCR system. The sensitivity was 0.0625ng DNA. Y-STR could be detection in a male/female DNA mixture ratio of 1:4. Conclusion The primary study demonstrates that this multiplex PCR system is effective and reliable for forensic routine DNA analysis. It will be very helpful for constructing Chinese forensic Y-STR database and population genetic research.