Objective To identify a rapid and efficient fungal genomic DNA extraction method for PCR amplification.Methods Genomic DNA was extracted from Penicillum marneffei,Rhizopus microsporus,Cryptococcus neoformans and Candida albicans by heating pyrolysis,microwave,repeated freezing and thawing,lysozyme digestion,overnight snail enzymatic and Qiagen kit methods.DNA electrophoretogram was observed by gel imaging system.The concentration and purity of extracted DNA were determined with an ultramicro nucleic acid protein tester and the yields were calculated.PCR amplification and sequencing were also performed.ANOVA and SNK-q test were used for data analysis.Results There were statistical differences in concentrations and yields of the fungal DNA extracted from Penicillum marneffei (hyphal phase and yeast phase),Rhizopus microsporus,Coptococcus neoformans and Candida albicans by six methods (F=750.83,220.95,669.35,132.01,510.20 and 1658.35,287.10,963.64,1147.77,4521.22,all P ＜0.01).Of six methods,microwave method gained the highest DNA concentration and yield,followed by heating pyrolysis method,while Qiagen kit method obtained the lowest concentration and yield.All DNA extracted by 6 kinds of methods were positive in PCR amplification.Conclusion All of the six methods can be used for fungal DNA extraction which is sufficient for PCR amplification,but microwave and heating pyrolysis methods are more easy and simple to perform.

Objective To investigate the epidemiological characteristics of mycoplasma pneumoniae (MP),Epstein-Barr virus (EB virus) and cytomegalovirus (CMV) infection in children with acute upper respiratory tract infections in Hangzhou.Methods Throat swabs and sputum samples were collected from 5942 children with acute upper respiratory tract infections in Hangzhou First People's Hospital during January 2011 and December 2012.MP,EB virus and CMV were detected using quantitative PCR.The distribution and seasonal changes of the above pathogens in children of different ages were analyzed using Chi-square tests.Results MP,EB virus and CMV were positive in 29.91％ (1777/5942),22.92％ (1362/5942) and 8.55％ (508/5942) children,respectively.Mixed infections were found in 556 (9.36％) children.The positive rates of MP varied among different age groups (x2 =113,P =0.000),and the highest one was detected in children ＞ 6-year old (448/1012,44.36％).EB virus infection was rare in age group 0-1 year,and the positive rate was of statistical difference from those in other age groups (x2 =167,181 and 187,P =0.000).The highest positive rate of CMV (23.78％) was found in children aged 0-1 year old.The positive rates of MP varied in different months of the year (x2 =208 and 211,P =0.000),and the highest positive rate was found in July and August.Conclusion The predominant pathogen of acute upper respiratory tract infection in children is MP in Hangzhou,and MP plus EB virus infection is common,particularly in older children;while CMV infection more likely occures in 0-1 year old babies,and usually in summer.

Xenotransplantation is one of the potential approaches to mitigate the shortage of donor kidneys. With the progress of gene modification techniques and the development of immunosuppressant, significant progress has been made in the preclinical research of genetically modified pig to non-human primate (NHP) xenotransplantation. The longest survival time of recipients exceeds 500 d. However, the number of recipients surviving for over 1 year is extremely low, and most recipients die within postoperative 1-2 months. Therefore, several problems remain to be clarified and resolved. In this article, rejection, refractory coagulation dysfunction, persistent inflammation, the selection of immunosuppressant, the selection of clinical recipients and the risk of cross-infection in genetically modified pig to NHP xenotransplantation were reviewed, and current problems and potential solutions of genetically modified pig to NHP xenotransplantation were summarized, aiming to provide reference for promoting xenotransplantation in clinical settings.

Objective:To explore the risk factors affecting the incidence of acute kidney injury(AKI)after liver transplantation(LT).Methods:From November 2019 to November 2022, clinical data were retrospectively reviewed for 105 recipients of classic orthotopic LT.There are 89 males and 16 females with an age range of(50.52±10.35)years.They are assigned into two groups of AKI(66 cases)and non-AKI(39 cases)according to the AKI diagnostic and staging criteria of Global Kidney Disease Prognosis Organization in 2012.General profiles and clinical data(e.g.previous medical history, MELD score, total bilirubin, albumin, serum creatinine level, coagulation function, anhepatic phase and time to surgery)of two groups of recipients are compared.The factors with statistically significant differences are included into multivariate Logistic regression analysis for obtaining independent risk factors for early AKI post-LT.Results:Among them, 66 patients developed AKI within 7 days post-operation with an incidence rate of 62.86%(66/105).The clinical stages of AKI are Ⅰ(46 cases, 69.70%), Ⅱ(10 cases, 15.15%)and Ⅲ(10 cases, 15.15%).Statistically significant inter-group differences exists in age, abdominal surgery history, preoperative serum level of creatinine, operative duration, anhepatic phase and intraoperative plasma transfusion(all P<0.05).Multivariate Logistic regression analysis indicated that abdominal surgery history( OR=5.803, 95% CI: 1.008~33.401, P=0.049), anhepatic phase( OR=1.054, 95% CI: 1.008~1.101, P=0.020)and preoperative serum level of creatinine( OR=0.968, 95% CI: 0.943~0.994, P=0.016)are independent risk factors for early AKI after classical orthotopic LT recipients. Conclusions:Abdominal surgery history, anhepatic phase, and preoperative serum level of creatinine are independent risk factors for early AKI in classic orthotopic LT recipients.

Objective To investigate the attitudes and influencing factors of transplantation-related populations towards kidney xenotransplantation. Methods From June 2022 to January 2023, stratified random sampling was performed from patients awaiting kidney transplantation, patients after kidney transplantation, patients' relatives and medical students. Four hundred subjects were collected from each population and 1600 subjects were investigated using a self-designed questionnaire. Baseline data of the respondents, their attitudes towards kidney xenotransplantation and the reasons of rejecting kidney xenotransplantation were analyzed. The influencing factors of attitudes towards kidney xenotransplantation were also identified. Results A total of 1 493 valid questionnaires were collected, and the questionnaire retrieval rate was 93.31%. About 93.10% of the respondents accepted allogeneic kidney transplantation, and 66.78% had heard of kidney xenotransplantation. Seven hundred and ninety-five respondents suggested that they could accept kidney xenotransplantation "when kidney xenotransplantation and allogeneic kidney transplantation yielded the same results and risks". Six hundred and ninety-eight respondents indicated that they were "unable" or "uncertain" whether they could accept kidney xenotransplantation (χ²=16.409，P=0.001). Among these 698 respondents, the proportion of them who were willing to accept kidney xenotransplantation when they did not meet the conditions of allogeneic kidney transplantation was 10.9%. About 35.8% of respondents were willing to accept kidney xenotransplantation if it yielded less risk and better prognosis compared with allogeneic kidney transplantation. If the time of awaiting kidney xenotransplantation was shorter than that of allogeneic kidney transplantation, 21.2% were willing to accept kidney xenotransplantation. If the cost of kidney xenotransplantation was less than that of allogeneic kidney transplantation, 24.5% of them were willing to accept kidney xenotransplantation. The main reasons of rejecting kidney xenotransplantation included surgical risk and other unknown risks. Multivariate analysis showed that respondents residing in cities and towns for a long period of time, those who accept allogeneic kidney transplantation and those who have heard of kidney xenotransplantation showed more positive attitudes towards kidney xenotransplantation. Conclusions Different transplantation-related populations have different attitudes towards kidney xenotransplantation, and the overall attitudes are positive. Active promotion of kidney xenotransplantation research and carrying out relevant popular science education contribute to improving public attitudes towards the acceptance of kidney xenotransplantation.