Feces and intestinal contents of pigs suspected with porcine epidemic diarrhea virus were collected from a farm in Minqin County,Gansu Province,China.After the suspected positive sam-ples were detected by RT-PCR,Vero cells were used to isolate and culture them in vitro.The suc-cessfully isolated virus was identified in the laboratory,and its whole genome sequence was ana-lyzed for genetic evolution.The pathogenicity was evaluated by animal regression test.The results showed that typical syncytial lesions could be observed when the PEDV-positive treatment solu-tion was inoculated with Vero cells in the 4th generation,and the virus titer in the 6th generation reached 10-4 75TCID50/mL.PEDV-like virions with a diameter of about 100 nm and a round shape with obvious capsular membranes and spikes were observed by electron microscopy.Whole genome sequencing analysis showed that the total length of this strain was 28 085 bp,which was far from the G1 subtype represented by the classical strain CV777(96.6％),and had a high homology with the G2b strains BC-2011-1,IA1,USA/Colorado/2013 and WELL(98.6％).This indicated that the strain belonged to the G2b epidemic strain.The animal regression test showed that the 5-day-old piglets developed vomiting,acute watery diarrhea,emaciation and mental depression within 12 h after the attack,and the symptoms worsened and died within 24 h.After autopsy,the infected piglets could be observed with stomach swelling,high intestinal heave,thin and transparent intesti-nal wall,and undigested milk clots inside.In summary,a PEDV G2b epidemic strain was success-fully isolated and identified in this study,and its whole genome sequence and pathogenicity were analyzed,providing research materials for future studies on PEDV gene function,pathogenic mech-anism and vaccine development.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

【Objective】 To investigate the effectiveness of multilink real-time fluorescence quantitative PCR (qPCR) in the detection of common pathogens in transplantation. 【Methods】 The primers of the qPCR detection system were designed for 24 common infectious pathogens after clinical transplantation, and the standard plasmids of each pathogen were used to verify the qPCR reaction.After the primer probe effect and concentration of each pathogen reaction system in this experiment was optimized, the sensitivity, correlation coefficient (R2) and amplification efficiency (E) of qPCR method were analyzed and confirmed.Twenty-two samples from patients, who underwent liver and kidney transplantation in transplant ICU of Sichuan Provincial People′s Hospital, were used to verify the application of the detection system.The total nucleic acid of 100 μL was extracted from each individual and divided into two aliquots, which were detected by multi-link qPCR reaction system and analyzed by high-throughput sequencing method (NGS). At the same time, samples (2 mL each) were taken from the transplanted patients for microbial culture.The results of the three detection methods were compared, and the NGS method was taken as the gold standard to analyze the positive detection rate of the multi-link qPCR method and its difference with the culture method and NGS. 【Results】 The lower limit of qPCR detection for 24 pathogens in the established qPCR detection system was 10¹cp/μL(R²>0.99), with the positive rate of pathogens at 59.1% (13/22), showing significant difference versus microbial culture (18.2%, 4/22)(P<0.05), but not versus NGS (63.6%, 14/22)(P>0.05). Percentage of pathogens detected was as follows: human herpetic virus type 6 (HHV-6) 30.8% (4/13), cytomegalovirus (HCMV) 23.1% (3/13), Epstein-Barr virus (EBV) 23.1% (3/13), human parvovirus B19 15.4% (2/13), Haemophilus influenzae (Hin) 15.4% (2/13), Enterococcus faecium (EFM) 15.4% (2/13), Clostridium difficile 15.4% (2/13), Escherichia coli 7.7% (1/13), Stenotrophomonas maltophilia (Sma) 7.7% (1/13), Klebsiella pneumoniae (Kpn) 7.7% (1/13), Enterococcus faecalis (Efa) 7.7% (1/13) and Streptococcus pneumoniae (Spn) 7.7% (1/13). The consistency rate of pathogens detected by the three methods was 32% (7/22), among which the consistency rate of multi-link qPCR with NGS method was 59% (13/22), and multi-link qPCR with microbial culture was 41% (9/22). 【Conclusion】 Compared with the microbial culture, the multi-link qPCR method demonstrated high sensitivity, accurate quantification, short time and low cost for the detection of common pathogens in clinical transplantation.Multi-link qPCR combined with NGS and microbial culture is helpful to quickly predict the pathogen infection status of patients after transplantation.

Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.

OBJECTIVE:To explore losses and gains (L&G) and L&G ratio induced by Essential Medicine System in a county. METHODS:By choosing a county in western China as sample area,field investigation was used to collect outpatient and inpatient visits,outpatient and inpatient income,drug income,total length of stay and medical insurance reimbursement criteria in primary medical institutions (township health centers,village health rooms) of the county during 2009-2015. By setting the year 2009 as the baseline year,the drug cost reimbursed by medical insurance was simulated and calculated when Essential Medicine System were not implemented;L&G and L&G ratio of medical insurance were calculated by comparing with actual drug cost reimbursed by medical insurance. RESULTS:The year 2012,in which the sample county fully implemented the Essential Medicine System was the turning year. Medical insurance funds lost in primary medical institutions of the county during 2010-2011(lost 437000,915000 yuan,respectively),but gained during 2012 to 2015(gained 199000,494000,858000,1290000 yuan, respectively);the L&G ratio increased from -0.67％ to 1.21％. For reimbursed outpatient drug cost and inpatient cost,L&G of medical insurance were different. For reimbursed drug cost of village health room and township health center,L&G of medical insurance were also different. CONCLUSIONS:The implementation of Essential Medicine System benefits to medical insurance within the county and Medical insurance funds can be saved.

Objective To investigate the correlation between tumor-associated macrophages (TAMs) and the development of high risk human papilloma virus (hr-HPV)-related cervical cancer. Methods A total of 112 cases of cervical tissue were collected, including 16 normal cervical tissues, 55 cervical intraepithelial neoplasia (CIN) tissues and 41 squamous cervical cancer (SCC) tissues. The expression of CD163+ macrophages in the cervical tissues was detected by immunohistochemical method, and the results were analyzed in relation with the clinical data of the patients. Results Immunohistochemical analysis showed that the cell density of CD163+macrophages increased progressively with the increase in the tissue malignancy, in the order of normal cervical tissue, CIN I, CIN II-III, and SCC. Correlation analysis revealed a positive relationship between CD163 + macrophage density and tissue malignancy (P=0.000). The density of CD163 + macrophages was significantly upregulated in HR-HPV-positive SCC tissue (P<0.05). CD163+ macrophages were positively correlated with cervical lymph node metastasis (P=0.005) and FIGO stage (P=0.004) of SCC. Conclusion The expression of CD163+macrophages is positively correlated with malignant transformation of cervical tissues, and hr-HPV infection is significantly correlated with CD163 expression level in the macrophages. CD163+ macrophages can be used as predictors of the occurrence and progression of cervical cancer caused by hr-HPV infection.

Objective To investigate the correlation between tumor-associated macrophages (TAMs) and the development of high risk human papilloma virus (hr-HPV)-related cervical cancer. Methods A total of 112 cases of cervical tissue were collected, including 16 normal cervical tissues, 55 cervical intraepithelial neoplasia (CIN) tissues and 41 squamous cervical cancer (SCC) tissues. The expression of CD163+ macrophages in the cervical tissues was detected by immunohistochemical method, and the results were analyzed in relation with the clinical data of the patients. Results Immunohistochemical analysis showed that the cell density of CD163+macrophages increased progressively with the increase in the tissue malignancy, in the order of normal cervical tissue, CIN I, CIN II-III, and SCC. Correlation analysis revealed a positive relationship between CD163 + macrophage density and tissue malignancy (P=0.000). The density of CD163 + macrophages was significantly upregulated in HR-HPV-positive SCC tissue (P<0.05). CD163+ macrophages were positively correlated with cervical lymph node metastasis (P=0.005) and FIGO stage (P=0.004) of SCC. Conclusion The expression of CD163+macrophages is positively correlated with malignant transformation of cervical tissues, and hr-HPV infection is significantly correlated with CD163 expression level in the macrophages. CD163+ macrophages can be used as predictors of the occurrence and progression of cervical cancer caused by hr-HPV infection.

Objective To explore the effect of fime particulate matters with an aerodynamic diameter less than 2.5 μtm (PM2.5) on daily cardiovascular disease mortality in seven cities of China.Methods Daily average concentrations of PM2.5,cardiovascular disease mortality data and environmental data were collected from January 1,2013 to December 31,2015 in seven cities of China,including Shijiazhuang,Haerbin,Shanghai,Wuhan,Guangzhou,Chengdu and Xi'an.We linked generalized additive model with Quasi-Poisson distribution to evaluate the association between daily concentrations of PM2.5 and cardiovascular disease mortality at single-city level and multi-city level,after adjusting for the long-term and seasonal trend,as well as meteorological factors and the effect of "days of week".Results The single-pollutant model indicated that there were marked differences in association strength in these cities,among which the effect in Guangzhou was strongest.At multi-city level,a 10 μg/m3 increase of PM2.5 was associated with an increase of 0.315％ (95％CI:0.133％-0.497％) of daily cardiovascular disease mortality.From lag0 to lag2,the effect of PM2.5 on cardiovascular disease mortality decreased,while it was strongest on lag01.In the two-pollutant model,the estimated effect decreased in all the cities with the adjustments of SO2 or NO2.The insignificant combined results suggested that PM2.5 might have combined effect with other pollutants.Each 10 μg/m3 increase of PM2.5 was associated with increases of 0.371％ (95％CI:0.141％-0.600％) and 0.199％ (95％ CI:0.077％-0.321％) of cardiovascular disease mortality in males and females,respectively.The effect of PM2.5 on cardiovascular disease mortality increased with age and decreased with educational level,although the differences between different subgroups were insignificant.The dose-response relationship between PM2.5 and cardiovascular disease mortality was non-linear and non-threshold,with a steeper curve at lower concentrations.Conclusion The increases of PM2.5 concentration can result in the increase of daily cardiovascular mortality.

Objective To investigate the correlation between carotid intraplaque neovascularization and acute ischemic cerebrovascular disease. Methods The patients examined with contrast-enhanced ultrasound were enroled and divided into either a symptomatic group or an asymptomatic group according to their cerebral ischemic symptoms. The patients were also divided into a low-echo group, an equal-echo group, and an heterogeneous echo group according to the plaque echo characteristics on conventional ultrasound. The carotid intraplaque neovascularization was evaluated with contrast-enhanced ultrasound. Multivariate logistic regression analysis was used to identify the correlation between carotid intraplaque neovascularization and acute ischemic cerebrovascular disease. Results A total of 73 patients with acute ischemic cerebrovascular disease were enroled, 32 patients in the symptoms group (19 ischemic stroke, 13 transient ischemic attack), 41 patients in the asymptomatic group. Plaque echo characteristics: low-echo 15, equal-echo 41, and heterogeneous echo 17. The proportions of the patients with plaque enhancement (84. 4% vs. 61. 0% ; χ2 = 4. 802, P = 0. 028) and enhanced intensity (21. 78 ± 8. 50 dB vs. 15. 93 ± 8. 82 dB; t = 2. 440, P = 0. 018) in the symptomatic group were significantly higher than those in the asymptomatic group. The proportions of the patients with enhanced plaque in the low-echo, equal-echo and heterogeneous echo group were 93. 3% , 58. 5% , and 82. 4% , respectively (χ2 = 7. 826, P = 0. 020 ). The low-echo group and heterogeneous echo group were significantly higher than the equal-echo group (al P < 0. 05), but there was no significant difference between the low-echo group and the heterogeneous echo group (P > 0. 05). The intraplaque enhanced intensities in the low-echo group, equal-echo group, and heterogeneous echo group were 22. 62 ± 9. 33 dB, 14. 38 ± 8. 02 dB, and 18. 15 ± 9. 64 dB, respectively (F = 3. 877, P = 0. 027). The low-echo group was significantly higher than the equal-echo group (P = 0. 024 ). Multivariate logistic regression analysis showed that carotid intraplaque neovascularization (odds ratio 3. 456, 95% confidence interval 1. 103 - 10. 828; P = 0. 033) was independently associated with acute ischemic cerebrovascular disease. Conclusions Carotid intraplaque neovascularization is closely associated with acute ischemic cerebrovascular disease.

BACKGROUND:Currently, hematopoietic stem celltransplantation mainly depends on unrelated donors. Mental state of the unrelated donors is very important to ensure the successful celltransplantation. OBJECTIVE:To compare mental and physical health status of relative and unrelated donors during the hematopoietic stem cellcol ection. METHODS:We compared the mental (Symptom Checklist-90) and physical (temperature, breath, pulse, and blood pressure) health status of relative and unrelated donors at admission, 1 day before cellcol ection, and 1-2 days after cellcol ection.RESULTS AND CONCLUSION:At admission, there was no difference in the mental health status of relative and unrelated donors (P>0.05), while the scores on Symptom Checklist-90 were significantly higher in the unrelated donors than the relative donors, including total score, forced, depression, anxiety, hostility, and fear (P<0.05). The physical signs were steady in the unrelated and relative donors, but the difference in breath and systolic blood pressure was of great significance before and after cellcol ection in the two groups. These findings indicate that during cellcol ection, the unrelated donors exhibit heavier mental load than the relative donors, and psychological counseling and health guidance are necessary.