Advances of environmental DNA technology in schistosomiasis surveillance / 中国热带医学

@#Abstract: Schistosomiasis, an important zoonotic parasitic disease, is one of the six major tropical diseases identified by WHO, and also one of the most important parasitic diseases for prevention and control in China. After more than 70 years of efforts, the prevention and control of schistosomiasis in China has made great achievements, and the current epidemic of schistosomiasis in China has entered an extremely low epidemic state, but the distribution base of the only intermediate host of schistosomiasis, Oncomelania hupensis, is still large. For now, the techniques used to monitor schistosomiasis have shortcomings such as time-consuming, laborious and low sensitivity, which cannot meet the current needs of China. Environmental DNA (eDNA) refers to DNA that can be extracted from environmental samples (such as soil, water or air) without isolating any target organisms, which is a complex mixture of genomic DNA and its degradation products from different organisms in the same environment. eDNA technology can reflect the community or species composition information in the ecosystem through DNA extraction and detection of environmental samples. Compared with traditional biological monitoring methods, eDNA technology has the advantages of high efficiency, high sensitivity and environmental friendliness. eDNA has been successfully used for the specific detection of Schistosoma mansoni, Schistosoma haematobium and Schistosoma japonicum. This paper reviews the current detection methods of eDNA, the application and technical limitations of eDNA technology in schistosomiasis monitoring, aiming to provide scientific reference for research in the field of schistosomiasis surveillance.

Analysis on the difference of circulating threshold of polymerase chain reaction in blood screening of external quality assessment / 中国输血杂志

【Objective】 To analyze the difference of circulating threshold (Ct) of polymerase chain reaction (PCR) in blood station laboratories during the external quality assessment, and to put forward suggestions for the quality improvement of participating laboratories. 【Methods】 From 2018 to 2021, the blood station laboratories participated in the external laboratory quality assessment of CITIC including blood screening items with nucleic acid testing method. The data of Roche diagnostic reagent group were used as the source, and the detected Ct values of three groups of quality control samples of HBV A subtype (400 IU/mL), HCV 1b subtype (400 IU/mL) and HIV B genotype (500 IU/mL) were used as the objects. The data were grouped according to quality control (sample) batches, reagent batches and different laboratories. Using the statistical method of variance analysis (assuming P<0.05 as significant), the detected Ct value of each group was analyzed. 【Results】 For the three items (HBV/HCV/HIV), the grouping data involving 42 batches of quality control (13/12/17), 28 batches of reagent (11/8/9) and 57 laboratories (19/19/19) were selected. The grouping analysis of quality assessment batches shows that there was no significant difference between HBV and HCV quality assessment batches, and there was no significant difference between other HIV batches except the two batches of HIV quality assessment samples released in 2021. The grouping analysis of each reagent batch showed that there was no significant difference between each reagent batch for HCV and HIV detection, while there was significant difference between two batches of HBV reagents. After excluding the data groups with significant differences in the quality control batch groups and the reagent batch groups, the detected Ct value of each laboratory group had extremely significant differences in the three items of HBV, HCV and HIV. Through pairing analysis, it was found that four laboratories had significant differences with most other laboratories in the three items, mainly manifested in the high mean value of Ct. 【Conclusion】 For the blood station laboratories with correct test results of quality assessment samples, there are differences in Ct values detected by PCR, which may be mainly caused by the detection ability of the participating laboratories.

HBV infection in window period among repeated blood donors: Residual risk assessment and trend analysis / 中国输血杂志

【Objective】 To investigate the status of blood safety and the effectiveness of preventive measures. 【Methods】 The data of Fengxian Blood Bank from 2018 to 2020 were extracted from Shanghai blood collection and supply information system. HBsAg sero-conversion samples of repeated blood donors were confirmed, and HBV serologic supplemental test were performed to obtain the number of new infections during the blood donation interval. The incidence and residual risk of HBV infection were evaluated by the sero-conversion model in donation intervals for repeated donors, and residual risk trend between the study period of 2002 to 2005, 2007 to 2011, 2011 to 2013 and 2018 to 2020 was compared. 【Results】 During 2018~2020, nine new HBV infections occurred among repeated donors during blood donation interval, with an incidence rate of 2.71 per 10 000. The residual risk of window period HBV transmission by transfusion could be reduced by 58.33% using HBsAg test plus NAT (HBsAg test 1∶30 637 vs HBsAg test plus NAT 1∶73 529). The residual risk of HBV transmission was decreasing when stratifying by periods, especially one order of magnitude dropped in 2018~2020 as in comparison of 2002 to 2005. 【Conclusion】 The residual risk of HBV transmission by transfusion showed a decrease trend. Although NAT could greatly reduce the risk, comprehensive preventive measures are needed to further reduce the risk.

External quality assessment of NAT blood screening: an inter laboratory analysis / 中国输血杂志

【Objective】 To summarize and analyze the results of local laboratories participating in China International Transfusion Infection Control (CITIC) Nucleic Acid Testing (NAT) External Quality Assessment (EQA). 【Methods】 The basic situation, test reagents, and abnormal results of 9 domestic laboratories participating in NAT EQA from 2018 to 2020 were collected and analyzed. 【Results】 Among 7 545 testing results, submitted by 48 laboratories using 8 test reagents throughout 9 occasions of CITIC, 64% (4 830/7 545) used imported and 36% (2 715/7 545), domestic reagents. Thirty-one abnormal results were reported, with false negative in 61.29% (19/31), false positive 6.45% (2/31), and others 32.25% (10/31). False negative results only appeared in samples with low viral load of HBV A genotype(40 IU/mL), HCV 1b subtype(40 IU/mL) and HIV B genotype(250IU/mL), relatively concentrated in a few laboratories. The frequency of abnormal results was 0.08 per laboratory per CITIC test. 【Conclusion】 The detection capacity of domestic blood stations has been significantly improved along with the routine NAT practice and regular NAT EQA participation over 5~10 years, but laboratory management still needs to be further strengthened to ensure the reagent testing performance and blood safety.

Treatment and pharmaceutical care of one patient with Chlamydia psittaci pneumonia complicated by drug-induced liver injury / 药学实践杂志

Objective To explore the role of clinical pharmacists in rational drug use through the pharmacy care of an elderly pneumonia patient with Chlamydia psittaci infection and drug-induced liver injury. Methods The clinical pharmacists participated in the treatment of one patient with Chlamydia psittaci pneumonia and drug-induced liver injury. Based on the results of second-generation gene sequencing, the characteristics of the pathogen were learned by literature search. The clinical pharmacists monitored the patient’s liver and kidney function, provided a new medication treatment plan to Doctors, and performed patient education during the treatment. Results The initial empirical anti-infective treatment with teicoplanin and imipenem-cilastatin was not effective. After the diagnosis of Chlamydia psittaci and Candida albicans infection, the combination of doxycycline with azithromycin and fluconazole was administered. Drug-induced liver injury was found with this treatment. The clinical pharmacist proposed to switch to doxycycline and clarithromycin with co-administration of magnesium isoglycyrrhizinate and polyene phosphatidylcholine to protect the liver. With this new regime, patient's liver function was improved and the infection was under control. Conclusion Individualized pharmaceutical cares provided by clinical pharmacists helped the safe, rational and effective use of medications.

The role of LL-37 in tumor development / 中国肿瘤生物治疗杂志

@# LL-37是人体内发现的唯一一种Cathelicidin类抗菌肽，由其前体hCAP-18经丝氨酸蛋白酶3剪切后产生。研究发现， LL-37可在卵巢癌、肺癌、恶性黑色素瘤、皮肤鳞状细胞癌、前列腺癌等肿瘤中发挥促癌作用；而在胃癌、结直肠癌、白血病等肿瘤 中发挥抑癌作用。本文就LL-37在这些恶性肿瘤发生发展过程中所起的作用作一综述。

Establishment of allele-specific PCR for the detection of CYP3A5 and MDR-1 gene polymorphisms and its clinical application / 临床检验杂志

Objective To establish an allele-specific PCR method for the detection of cytochrome P-450 CYP3A5 (A6986G) and multiding resistance gene MDR-1 (C3435T) polymorphisms,and investigate the correlations of their polymorphisms with blood tacrolimus (Tac) concentration/dose (C/D) ratio in renal transplant recipients.Methods The allele-specific PCR primers were designed according to the polymorphism sites of CYP3A5 (A6986G) and MDR-1 (C3435T) genes.Then,their polymorphisms in the genomic DNA of peripheral blood samples from 72 renal transplant recipients were analyzed,and the results were validated by DNA sequencing.The blood Tac concentration was determined by the chemiluminescence microparticle immunoassay and the differences of concentration,dose and C/D ratio of blood Tac in renal transplant recipients with different genotypes were compared at 1 month after transplantation.Results The coincidence rate between the established allele-specific PCR and DNA sequencing was 100％.The frequencies of CYP3A5 * 1/* 1,* 1/* 3 and * 3/* 3 genotypes in 72 renal transplant recipients were 18.1％,31.9％ and 50.0％,respectively,and those of MDR-1 C/C,C/T and T/T genotypes were 27.8％,58.3％ and 13.9％,respectively.There were significant differences in blood Tac concentration (P =0.014) and Tac C/D ratio (P =0.019) between different CYP3A5 genotypes of renal transplant recipients.Further analysis found that the Tac C/D ratio of CYP3A5 * 3/* 3 genotype was significantly higher than that of CYP3A5 * 1/* 1 and * 1/* 3 genotypes (P ＜ 0.05).Conclusion The allele-specific PCR method for the detection of CYP3A5 and MDR-1 polymorphisms is successfully established and the polymorphism of CYP3A5 * 3 gene in renal transplant recipients is obviously correlated with the pharmacokinetics of Tac.

The study on nucleic acid testing for bacterial detection and its preliminary application on concentrated platelet / 中国输血杂志

Objective To establish a nucleic acid testing assay for bacterial detection with meeting automatic requirement,and use it preliminarily to detect bacterial contamination in concentrated platelet.Methods Optimizing the bacterial DNA extraction methods,evaluating the sensitivity and specificity of the optimized assay with different bacterial strains,and applying the assay on bacterial detection for 100 units of concentrated platelet.Results The optimized extraction methods met the criteria of automatic testing.The sensitivity of the assay is 10,150,65,15 CFU/mL for Escheriehia coli,Stphylococcus aureus,Bacillus subtilis,and Pseudomonas aeruginas respectively.There was no non-specific amplification detected.And the assay could detect 1 contaminated bacterial from 100 units of concentrated platelet as same as BactALT bacterial culture.Conclusions Nucleic acid testing for bacterial genome 16S DNA could detect common contaminated bacterial sensitively and fast from platelet product.This assay could have the same effect when compared to bacterial culture methods when it was applied in concentrated platelet products.

The clinical and pathological features of patients with infective endocarditis diagnosed at autopsy / 中华内科杂志

Objective To study the characteristics and misdiagnosis of infective endocarditis (IE).Methods Clinical and pathological data of 15 IE patients diagnosed by autopsy were collected,compared with the cohort of 29 regular IE cases.Results In the autopsy patients,IE in the left heart was predominat (14/15,93.3％).Arterial embolism was the most common comorbidity (10/15,66.7％),followed by pneumonia,malignancies,pyelonephritis (9/15,60.0％;7/15,46.7％;6/15,40.0％ respectively).The age was elder [(64.0 ± 21.2) years vs (50.8 ± 18.3) years,t =-2.15,P =0.037] in autopsy patients than in control group.More patients of missed diagnosis were combined with malignancies or deep vein catheterization [7/15 (46.7％) vs 1/29 (3.4％);5/15 (33.3％) vs 0/29 (0) respectively].Fever and cardiac murmur were rare in misdiagnosed cases [11/15 (73.7％) vs 28/29 (96.9％),0/15 (0) vs 15/29 (51.7％) respectively].Echocardiography and blood culture were performed in only 33.3％ (5/15) and 26.7％ (4/15) cases of missed diagnosis with low positive rates compared with regular IE patients [0/15 (0) vs 27/29 (93.1％);1/15(6.7％) vs 15/29(55.6％)].Conclusions Infective endocarditis should be suspected in patients with the risk factors of IE even without fever or cardiac murmur.Echocardiography and blood culture should be done as screening tests of IE.

The clinical and pathological features of patients with infective endocarditis diagnosed at autopsy / 中华内科杂志

Objective To study the characteristics and misdiagnosis of infective endocarditis (IE).Methods Clinical and pathological data of 15 IE patients diagnosed by autopsy were collected,compared with the cohort of 29 regular IE cases.Results In the autopsy patients,IE in the left heart was predominat (14/15,93.3％).Arterial embolism was the most common comorbidity (10/15,66.7％),followed by pneumonia,malignancies,pyelonephritis (9/15,60.0％;7/15,46.7％;6/15,40.0％ respectively).The age was elder [(64.0 ± 21.2) years vs (50.8 ± 18.3) years,t =-2.15,P =0.037] in autopsy patients than in control group.More patients of missed diagnosis were combined with malignancies or deep vein catheterization [7/15 (46.7％) vs 1/29 (3.4％);5/15 (33.3％) vs 0/29 (0) respectively].Fever and cardiac murmur were rare in misdiagnosed cases [11/15 (73.7％) vs 28/29 (96.9％),0/15 (0) vs 15/29 (51.7％) respectively].Echocardiography and blood culture were performed in only 33.3％ (5/15) and 26.7％ (4/15) cases of missed diagnosis with low positive rates compared with regular IE patients [0/15 (0) vs 27/29 (93.1％);1/15(6.7％) vs 15/29(55.6％)].Conclusions Infective endocarditis should be suspected in patients with the risk factors of IE even without fever or cardiac murmur.Echocardiography and blood culture should be done as screening tests of IE.

Preliminary histological and biomechanical study about the timing of surgical repair for acute rotator cuff tears in rabbits / 中华外科杂志

<p><b>OBJECTIVES</b>To establish the animal model of acute rotator cuff tear in rabbits, and study the effect of timing of surgical repair on healing of tendon-bone interface, formation and distribution of collagens in the supraspinatus tendon insertion and biomechanical properties of supraspinatus.</p><p><b>METHODS</b>Supraspinatus tenotomy was performed in the right shoulder of 90 skeletally matured male New Zealand white rabbits to establish the animal model of acute rotator cuff tear. The rabbits were randomly divided into 3 groups : group of early repair, repaired at 1 week after tenotomy; group of late repair, repaired at 4 weeks after tenotomy; and group without repair, used as control. At 2 weeks, 4 weeks and 8 weeks after repair, healing of tendon-bone interface was observed by HE staining. Collagens were observed by Sirius Red F 3B (SR) in saturated carbazotic acid staining. The areas of type I and III collagens were measured by using imaging analysis software and the ratio of type I and III collagens were calculated. Failure loads of supraspinatus on both sides were measured. The percentage of failure loads of the surgical side was calculated and contralateral supraspinatus were uninjured.</p><p><b>RESULTS</b>There was no obvious fatty infiltration and muscle atrophy in supraspinatus in all groups. At 8 weeks, the formation of a new enthesis of supraspinatus in groups of early and late repair were observed. In groups of early and late repair, the ratio of areas of type I and III collagens at 8 weeks (2.02 ± 0.77 and 2.06 ± 0.58) was larger than that at 2 weeks (1.10 ± 0.24 and 1.14 ± 0.50, t = 3.082, 3.655, P < 0.01). At 2, 4 and 8 weeks, the percentages of failure loads of the surgical side and uninjured contralateral supraspinatus in group of early repair(38% ± 11%, 66% ± 7%, 89% ± 4%) and group of late repair (41% ± 16%, 63% ± 7%, 89% ± 9%) were both higher than that in group without repair (14% ± 6%, 32% ± 4%, 56% ± 12%); the differences were all statistically significant (group of early repair: t = 3.311, 8.549, 5.719; group of late repair: t = 3.713, 8.063, 6.044; P < 0.01). The percentage of failure loads of the surgical side and uninjured contralateral supraspinatus at 8 weeks was higher than those at 4 weeks (t = 3.878 - 4.613, P < 0.01) and 2 weeks (t = 7.158 - 10.024, P < 0.01) in all groups.</p><p><b>CONCLUSIONS</b>Surgical repair within 4 weeks of acute rotator cuff tear lead to formation of a new enthesis of supraspinatus, improvement of both ratio of type I collagen in the supraspinatus tendon insertion and biomechanical properties of supraspinatus.</p>

Expression of translationally controlled tumor protein in human breast cancer and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of translationally controlled tumor protein (TCTP) in human breast cancer tissues and its clinical significances.</p><p><b>METHODS</b>The expression of TCTP in 94 human breast cancer and the corresponding adjacent normal mammary tissues were detected using immunohistochemistry.</p><p><b>RESULTS</b>The expression rate of TCTP was 64.89% in human breast cancer tissues, significantly higher than that in normal benign mammary tissues (39.36%, P<0.001). TCTP overexpression was positively correlated to the tumor size, clinical stage, lymph node metastasis and histological grade of breast cancer (P<0.05). Patients with positive TCTP expression had a significantly shorter mean survival time than those with negative expression (P<0.001).</p><p><b>CONCLUSION</b>TCTP may play an important role in the tumorigenesis and development of breast cancer, and can be an important prognostic factor for this malignancy.</p>

Joint application of mathematic models in assessing the residual risk of hepatitis C virus transmitted through blood transfusion / 中华流行病学杂志

The practicable and effective methods for residual risk assessment on transfusion-transmitted disease was to establish the mathematic models. Based on the characteristics of the repeat donors which donated their blood on a regular base, a model of sero-conversion during the interval of donations was established to assess the incidence of the repeat donors. Based on the characteristics of the prevalence in the population, a model of ‘prevalence increased with the age of the donor' was established to assess the incidence of those first-time donors. And based on the impact of the windows period through blood screening program, a model of residual risk associated with the incidence and the length of the windows period was established to assess the residual risk of blood transfusion. In this paper, above said 3 kinds of mathematic models were jointly applied to assess the residual risk of hepatitis C virus (HCV) which was transmitted through blood transfusion in Shanghai,based on data from the routine blood collection and screening program. All the anti-HCV unqualified blood donations were confirmed before assessment. Results showed that the residual risk of HCV transmitted through blood transfusion during Jan. 1st,2007 to Dec. 31st,2008 in Shanghai was 1∶101 000. Data showed that the results of residual risk assessment with mathematic models was valuable. The residual risk of transfusion-transmitted HCV in Shanghai was at a safe level, according to the results in this paper.