EGFR mutation, expression of PD-L1/Ki-67 and their correlation with clinical features in elderly NSCLC patients / 中华检验医学杂志

Objective:To investigate the mutation of epidermal growth factor receptor (EGFR), the expression of programmed death ligand 1 (PD-L1), cell proliferation-associated antigen (Ki-67) in elderly patients with non-small cell lung cancer (NSCLC), and their correlation with clinical feature such as gender, histological type and TNM stage.Methods:The tissue samples of 340 elderly NSCLC patients with definite histopathological diagnosis were collected from January 2020 to December 2020 in Huadong Hospital Affiliated to Fudan University, including 195 males and 145 females, age between 68.9±6.0 years. Patients were grouped according to clinical features such as gender, histological type and TNM stage. The expression of EGFR mutation, PD-L1 and Ki-67 were detected by Super-ARMS and immunohistochemistry. The correlation between tnem and clinical features was statistically analyzed, and the correlation between EGFR mutation and PD-L1/Ki-67 expression was further analyzed separately.Results:In elderly NSCLC patients′ tissues, the positive rate of EGFR mutation was 48.53% (165/340). L858R and 19del mutations were the most common types, which were 56.36% (93/165), 30.30% (50/165) respectively. The mutation rate of EGFR was higher in women, lung adenocarcinoma, well-differentiated, and low-stage patients, which were 65.52% (95/145), 53.77% (164/305), 56.75% (143/252), 52.53% (135/257) respectively. In addition, the positive rate of PD-L1 expression was higher in elderly patients with non-adenocarcinoma lung cancer and poorly differentiated adenocarcinoma, which were 37.14% (13/35), 24.53% (13/53) respectively. The negative rate of PD-L1 expression was higher in elderly patients with NSCLC in stage Ⅰ+Ⅱ, no lymph node metastasis and weakly positive Ki-67, which were 89.11% (229/257), 87.63% (248/283), 94.71% (197/208) respectively. Correlation analysis showed that EGFR mutation was negatively correlated with the expression of PD-L1 and Ki-67 (PD-L1: r=-0.22, P<0.001; Ki-67: r=-0.32, P<0.001). Conclusion:There is a negatively correlation between EGFR mutation and the expression of PD-L1 and Ki-67 in elderly NSCLC, suggesting that the combined detection of EGFR mutation and PD-L1 expression could provide the basis for precise targeted therapy for elderly NSCLC patients.

Potential diagnostic and therapeutic targets and anti-tumor mechanisms of Neddylation pathway in digestive system tumors / 中华检验医学杂志

Neddylation,a novel post-translational modification of proteins, is overactivated in digestive system tumors and can be used as a potential anti-tumor molecular target. Targeting Neddylation pathway plays an anti-tumor role by inducing cell cycle arrest, apoptosis, senescence and autophagy of digestive system tumor cells, as well as enhancing the sensitivity of digestive system tumor cells to the radiotherapy and chemotherapy. Targeting Neddylation pathway and its inhibnitor MLN4924 can act as poential targets against digestive system tumors.

Social net work analysis of referral of stroke inpatients in Changsha / 中华医院管理杂志

Objective:To analyze the current status of referral of stroke inpatients and explore the characteristics of the referral network in Changsha, for the reference to improve and promote the hierarchical medical system of stroke.Methods:Data of the inpatient medical record of stroke patients and the annual reports of medical institutions in Changsha in 2018 were collected from the health statistics network direct reporting system of Health Commission of Hunan province, for analysis of the referrals of inpatients in different medical institutions. Social network analysis was adopted to analyze the density, centrality and K-core of the referral network of stroke inpatients.Results:A total of 82 medical institutions for stroke inpatients were included with 2 859 referrals of patients. Most of the referrals were made between tertiary hospitals(1 515), especially within hospitals of a stroke alliance(1 123). The density of referral network was 0.613.Tertiary hospitals were in the center of the network, the entry points of secondary hospitals were in the center of the network and primary medical institutions were located at peripheral positions. Most of the tertiary hospitals in the 15-core(14, 72.68%), 12 of them were the units of Hunan Stroke Alliance.Conclusions:Tertiary hospitals played an important role in the region, secondary hospitals were able to receive patients referred by tertiary hospitals, but few patients were transferred to primary care institutions; The primary medical institutions failed to play due roles in the referral network. The establishment of stroke alliances could promote the cooperation of hospitals in the alliance, but the division of labor and cooperation among different levels of medical institutions in the region needed to be further optimized.

Research on the anti-lung cancer effects of targeted Neddylation modifying pathway and its inhibitor MLN4924 and its mechanism / 国际肿瘤学杂志

Neddylation is overactivated in lung cancer, which promotes the development of lung cancer by activating its downstream CRL ubiquitin ligase and promoting the CRL tumor-suppressor protein substrate degradation. MLN4924, a small molecule inhibitor of Neddylation, plays an anti-lung cancer role by inducing cell cycle arrest, apoptosis and senescence. Furthermore, targeting the key enzymes of Neddylation and their substrates, Cullin family proteins, can inhibit the development of lung cancer.

Progress of cell death induced by Neddylation modification pathway inhibitor MLN4924 / 肿瘤研究与临床

Protein ubiquitin-like modification of Neddylation is an important post-translational modification mode of protein to widely regulate cell cycle, growth, development and other biological processes. Recent studies have found that MLN4924, small molecule inhibitor of Neddylation modification, has a significant anti-tumor effect, which can inhibit the tumor growth by inducing cell cycle arrest, senescence and apoptosis, and inhibiting tumor angiogenesis. This paper reviews the related mechanism research of cell programmed death induced by Neddylation modification pathway inhibitor MLN4924.

Establishment, evaluation, and determination of saliva glucose concentration by ion chromatography / 中华内分泌外科杂志

Objective:To establish an analytical method for measuring the concentration of glucose in saliva by ion chromatography.Methods:The proteins in saliva were removed by thermal denaturation method, CarboPac PA20 (3×30 mm) was used as a protective column and CarboPac PA20 (3×150 mm) was used as an analytical column for ion chromatography analysis. Gradient elution was carried out with A: ultra-pure water, B: 250 mmol/L NaOH solution and C: 500 mmol/L NaAc solution. Pulsed ampere detector was used for detection.Results:This method had a good linear relationship in the range of 0.04 to 0.12 mg/L, with a linear relation coefficient of 0.9967. The detection limit of glucose was 2 μg/L, the mean value of the relative standard deviation (RSD) of the repeatability measurement was 0.75%, and the average spike recovery was 103.07%.Conclusion:This method is simple, sensitive, accurate and stable, and can be used for the determination of glucose concentration in saliva.

Clinical application of circulating tumor cells in the diagnosis and treatment of urothelial carcinoma / 中华检验医学杂志

Objective:To study the correlation between circulating tumor cells (CTC) and the degree of pathological invasion, recurrence and metastasis of urothelial carcinoma, and so to explore the clinical value of CTC detection in bladder cancer.Methods:A total of 142 patients with urothelial carcinoma in Huadong Hospital Affiliated to Fudan University were enrolled as cancer group from July 2016 to January 2018. According to the degree of tumor invasion, cancer group was divided into the non-muscle-invasive group (49 cases) and the muscle-invasive group(93 cases). In addition, 52 patients with benign urinary tract lesions admitted were selected as the benign group and 56 patients with non-urinary tract diseases and non-tumor as the control group. A total of 3.2 ml of venous anticoagulant blood from each subject was collected. CTC was enriched by negative enrichment using the magnetic beads coated with monoclonal antibody Cluster 45 of differentiation (CD45) to capture and remove white blood cells, and identified by chromosome 8 probe(CEP8) fluorescence in situ hybridization (FISH) technique. CD45-/4′,6′-diamidino-2-phenylindole+/CEP8>2(CD45-/DAPI+/CEP8>2) cells were judged as CTC. SPSS22.0 statistical software was used for statistical analysis.Results:≥2 CTCs/3.2 ml in blood was set as cutoff value. CTC positive rates in bladder cancer group, benign group and control group were 70.42%(100/142), 28.85%(15/52) and 8.93%(5/56), respectively, and there was a significant difference (χ 2=70.496, P=0.000). There was a statistically difference ( U=2 863.5, P=0.011) in the mean count of CTC(2 CTCs/3.2 ml vs 4 CTCs/3.2 ml) between the two groups. The proportion of≥5 CTCs/3.2 ml in the muscle-invasive group was 40.86% (38/93), which was significantly higher than that in the non-muscle-invasive group, 18.37% (9/49) (χ 2=7.330, P=0.007). Cystoscope follow-up of 65 patients treated with transurethral resection of the bladder tumor showed that the recurrence and metastasis rate in patients with≥5 CTCs/3.2 ml was as high as 47.62% (10/21), compared with 11.36% (5/44) of patients with<5 CTCs/3.2 ml (χ 2=10.530, P=0.001). Among 59 patients undergoing radical cystectomy, no significant difference was found in tumor diameter >3 cm, positive surgical margins and positive lymph nodes among all groups according to CTC negative or positive and CTC number ( P>0.05). But the recurrence and metastasis rate of patients with ≥5 CTCs/3.2 ml (59.10%) was significantly higher than that of patients with <5 CTCs/3.2 ml (6/30)(χ 2=8.364, P=0.004). Conclusion:The number of CTC increased with the deepening of tumor invasion; Tumor recurrence and metastasis increased significantly in the patients with ≥5/3.2 ml CTCs in blood.

Methods of packaging lentivirus library for CRISPR/cas9 -based screening / 中华实验和临床病毒学杂志

Objective@#To obtain the optimum of lentiviral library packaging based on CRISPR/cas9 (Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9).@*Methods@#Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence antibody (IFA) and enzyme linked immunosorbent assay (ELISA) were used to detect the lentivirus titers in condition of different ratio of packaging plasmids, different addition of lipofectamine 3000 reagent and different time points post-transfection. Then, high-throughput sequencing was performed to evaluate the representation and distribution of single guide (sg)RNAs in the library.@*Results@#The lentivirus titer was the highest when the molar ratio of psPAX2∶pMD2.0G∶Lentivirus library was 2∶1∶1, and the optimum addition of Lipofectamine 3000 reagent was 10 μl, while the result of ELISA were correspondent to that of RT-PCR. The IFA result showed that the lentivirus titer was the highest at 60 h post-transfecion. The coverage of sgRNAs in the lentivirus library packaged with the optimum we obtained was 99.3%, and the read counts of sgRNAs was observed in a normal distribution.@*Conclusions@#The optimal lentivirus library packaging was obtained, and this can provide basis for CRISPR/cas9-based screening.

Clinical application of circulating tumor cells in the diagnosis of prostate cancer / 中华检验医学杂志

Objective To evaluate the diagnostic value of circulating tumor cells(CTCs) in prostate cancer (Pca) through studying the relationship between CTCs and Gleason scores and pathological TNM stage in Pca patients. Methods A total of 238 patients including 161 Pca patients as cancer group, 35 male patients with benign prostatic diseases as benign group and 42 male with non-prostate disease as control group, who were treated in our hospital from July 2016 to January 2018,were enrolled. Venous blood of every patient was collected and CTCs were enriched and identified by immunocytochemistry CD45 capturing leukocyte and fluorescence in situ hybridization with chromosome 8 (CEP8-FISH). Cells displaying CD45-/DAPI+/CEP8>2 were characterized as CTCs. One-way ANOVA was used to exam the correlations of the number of CTCs with Gleason scores and pathological TNM stage. Results CTCs ≥2 were detected in 74.53％(120/161) of Pca patients and 20.00％(7/35)of benign prostatic diseases patients and 7.14％(3/42)of control group (χ2=79.605,P<0.05). In group Gleason scores 6, the numbers of CTCs were 2.00 ± 2.42, the ratios of CTCs≥5 and tetraploid were 13.33％ (2/15)and 26.67％(4/15) respectively. In 7 scores group, the results were 3.14±2.68,17.72％(14/79) and 34.18％(27/79)respectively;In 8 scores group, the results were 3.57 ± 2.70, 33.33％(7/21)and 42.86％ (9/21)respectively; In 9 scores group, these three results were 4.65±4.41, 43.48％(20/46) and 45.65％(21/46)respectively. The numbers of CTCs in the≤pT2b (20), pT2c(27), pT3a(19), pT3b(16)and≥pT4(12) groups were 2.25±2.45, 3.56±2.79, 4.05±3.47, 4.69±2.12 and 5.17±3.21 respectively. The ratios of CTCs≥5 were 25.00％(5/20), 25.93％(7/27), 26.32％(5/19), 50.00％(8/16) and 58.33％ (7/12)respectively. The proportions of tetraploid were 20.00％(4/20), 25.93％ (7/27), 31.58％(6/19), 50.00％(8/16) and 58.33％(7/12) respectively. There were significant differences between CTC and Gleason scores (F=3.200, P<0.05)and pathological stage (F=2.673, P<0.05). The ratios of CTCs≥5 increased with the increase of Gleason scores (χ2=11.592, P<0.05). Conclusions The detection of CTCs could be used for the differential diagnosis of Pca and benign prostatic disease. There were notable correlations between the numbers of CTCs and Gleason scores and pathological stage in Pca patients, especially between CTCs≥5 and Gleason scores.

Epidemiological characteristics of tick borne encephalitis in China, 2014 / 中华实验和临床病毒学杂志

Objective@#To analyze the epidemiological characteristics and distribution characteristics of tick-borne encephalitis in China in 2014, and to provide scientific basis for formulating specific prevention and control measures.@*Methods@#The epidemic data were obtained from the "infectious disease report information management system" , using Excel 2016, GIS and other software to summarize and analyze the cases of tick borne encephalitis (TBE) reported, using the number of cases, incidence, composition ratio and other indicators to analyze and describe the TBE epidemiological characteristics in China in 2014.@*Results@#In 2014, a total of 322 cases of TBE were reported in 9 provinces in China, with an annual incidence of 0.024/100, 000 and 1 death of patient. The provinces with high number of cases were Jilin province, Inner mongolia autonomous region and Heilongjiang province, and the number of cases in the other six provinces is no more than two. TBE was distributed in spring and summer, and it is concentrated in May to July. The age of the affected population was mostly concentrated in 40-49 years old, the male-female ratio was 1.6∶1 (198/124), and the patients were dominantly farmers, household and unemployed workers, and forestry workers, they accounted for 49.40% (159/322), 26.40% (85/322) and 18.60% (60/322) of the national TBE cases respectively. The three hospitals that reported the most TBE cases in 2014 were Inner mongolia forestry general hospital, Jiangyuan People′s hospital of Baishan city, Jilin province and Mudanjiang forestry central hospital of Heilongjiang province. The number of reported cases in these three hospitals accounted for 68.6% of the whole country. The laboratory diagnosis rate of Inner mongolia forestry general hospital was the highest (91.9%).@*Conclusions@#In 2014, the incidence of TBE in China has continued to rise compared with the previous two years. The geographical focus is mainly on the forest areas of Daxing′anling, Xiaoxing′anling and Changbai Mountain.

Inhibitory effect of andrographolide on angiogenesis induced by the supernatant from cultured tumor cells / 中南大学学报(医学版)

To determine the effect of andrographolide (Andro) on angiogenesis of human umbilical vein endothelial cells (HUVECs).
 Methods: HUVECs were treated with different concentrations of Andro and the cell viability was detected with Cell Counting Kit-8 (CCK-8). HUVECs were treated with half lethal dose (IC50) of Andro. Matrigel was used to make capillary formation of HUVECs and the effect of Andro on capillary formation was evaluated by calculating the percentage of capillary formation. Moreover, the effects of Andro and the supernatant from cultured A549 tumor cells on capillary formation were evaluated by calculating the percentage of capillary formation. The effect of Andro on the expression of matrix metalloproteinase-9 (MMP-9) was determined with Western blot.
 Results: The cell viability of HUVECs decreased with the increase of Andro concentrations. IC50 was 20 μmol/L. The capillary formation of HUVECs was inhibited when treated with 20 μmol/L Andro for 24 hours. Moreover, Andro was able to antagonize the promotion of the capillary formation induced by the supernatant from cultured tumor cells. Andro could suppress the expression of MMP-9 and antagonize the capillary formation.
 Conclusion: Andro inhibits the capillary formation of HUVECs and can antagonize the promotion of angiogenesis induced by the supernatant from cultured tumor cells.

Application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry for the ;identification of Helicobacter pylori / 中华检验医学杂志

Objective To optimize the pre-treatment method before detecting Helicobacter pylori ( H.pylori ) by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry ( MALDI-TOF MS) and evaluate the ability of the Superspectra by MALDI-TOF MS for identifying clinical isolated H.pylori strains.Methods H.pylori were isolated from 469 biopsy samples of gastric mucosa collected from January 2015 to July 2016 in Huadong Hospital affiliated to Fudan University , 16 s rRNA sequencing were then performed to validate the strains.Then 91 isolated H.pylori strains were used for the subsequent MALDI-TOF analysis.The effect of pre-treatment of 50%isopropanol, formic acid and acetonitrile (1∶1), 70%formic acid were compared before H.pylori detecting by MALDI-TOF MS.40 out of 91 clinical H.pylori strains were detected by MALDI-TOF MS and the spectra were randomly assigned to 4 groups including 5, 10, 20, 30 spectra, each group had 3 repeats.Then 4 groups with different amount of spectra were used for creating Superspectra with SARAMIS Premium software , respectively.The remaining 51 H.pylori strains including 306 spectra were used for validating the identification rate of the Superspectra.Results With the use of 70%formic acid, the greater number of ion signals and higher relative intensity of the main peaks were observed than other pre-treatment reagents.The identification rate of Superspectra created by 30 strains group was the highest ( 90.2%).Among the 306 spectra, 46.1% of them achieved highly reliable identification , 22.2% achieved lower degree of reliable identification , and 31.7% of them achieved “no identification”.Conclusions The study optimized the pre-treatment method before H.pylori detecting by MALDI-TOF MS.The Superspectra was created with the good ability to rapidly identify clinical isolated H.pylori strains.

Digitalization of information resources in domestic TCM / 中华医学图书情报杂志

After the following aspects of information resources digitalization in domestic TCM were described, in-cluding different types and contents of digital information resources, effectively developed ancient book resources, expanded team construction, and strengthened cooperation between departments. Measures were put forward for the digitalization of information resources in domestic TCM according to the existed problems, such as insufficient de-velopment, lack of audiovisual products, and low accessibility of TCM information resources.

Serological investigation of vector-borne disease in dogs from rural areas of China

Objective:To evaluate the Anaplasma phagocytophilum (A. phagocytophilum), Ehrlichia canis (E. canis), Dirofilaria immitis (D. immitis) (canine heartworm), Borrelia burgdorferi (B. burgdorferi) infections in countryside dogs from Yunnan, Hainan and Anhui provinces. Methods: Serum samples were collected from 26 dogs in Yunnan, Hainan and Anhui provinces. The samples were tested using a commercial ELISA rapid diagnostic assay kit (SNAP? 4Dx?; IDEXX Laboratories, Inc. U.S.A.). Meanwhile, indirect immunofluorescence assay (IFA) recommended by WHO was conducted to detect IgG to A. phagocytophilum. Two methods were analyzed and compared. Results: The number of serologically positive dogs for IgG to A. phagocytophilum was only 2 which was from Hainan province and none of the 26 dogs responded positive for E. canis, D. immitis (canine heartworm), and B. burgdorferi by ELISA rapid diagnostic method. The number of serologically positive dogs for IgG to A. phagocytophilum was 13 (50%) by IFA method. Data of the two methods were analyzed by statistical software and the difference was statistically significant (P=0.002). Conclusions: It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method. However, we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including A. phagocytophilum, E. canis, D. immitis (canine heartworm), and B. burgdorferi which have public health significance.

Mutational analysis of KCNJ11 in Chinese elderly essential hypertensive patients / 老年心脏病学杂志（英文版）

Objective To compare the distribution of KCNJ11 polymorphisms between elderly Chinese population with and without hypertension. Methods We examined the mutation of KCNJ11 gene by directly sequencing. Data for the present study were obtained from 250 hypertensive subjects (60 to 83 years old) as well as 250 normotensive subjects (60 to 86 years old). Results We found nine different mutations in KCNJ11, including six novel mutations (I131M, L147I, L147V, L147L, Q235H, G245C). None of the novel mutations were found in the normotensive subjects, and all the residues were conserved in other species. These sequence variants in Chinese population indicate the diversity of the human library and the complexity of hypertension. Conclusions The consistent finding of our present study provided a basis for the development of new strategies to diagnosis and treat hypertension in the elderly.

Serological investigation of vector-borne disease in dogs from rural areas of China

<p><b>OBJECTIVE</b>To evaluate the Anaplasma phagocytophilum (A. phagocytophilum), Ehrlichia canis (E. canis), Dirofilaria immitis (D. immitis) (canine heartworm), Borrelia burgdorferi (B. burgdorferi) infections in countryside dogs from Yunnan, Hainan and Anhui provinces.</p><p><b>METHODS</b>Serum samples were collected from 26 dogs in Yunnan, Hainan and Anhui provinces. The samples were tested using a commercial ELISA rapid diagnostic assay kit (SNAP(®) 4Dx(®); IDEXX Laboratories, Inc. U.S.A.). Meanwhile, indirect immunofluorescence assay (IFA) recommended by WHO was conducted to detect IgG to A. phagocytophilum. Two methods were analyzed and compared.</p><p><b>RESULTS</b>The number of serologically positive dogs for IgG to A. phagocytophilum was only 2 which was from Hainan province and none of the 26 dogs responded positive for E. canis, D. immitis (canine heartworm), and B. burgdorferi by ELISA rapid diagnostic method. The number of serologically positive dogs for IgG to A. phagocytophilum was 13 (50%) by IFA method. Data of the two methods were analyzed by statistical software and the difference was statistically significant (P=0.002).</p><p><b>CONCLUSIONS</b>It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method. However, we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including A. phagocytophilum, E. canis, D. immitis (canine heartworm), and B. burgdorferi which have public health significance.</p>

The effects of mycophenolate mofetil on immunologic compromise in rats with severe acute pancreatitis / 中华急诊医学杂志

Objective To observe the modulatory effects of NF-κB on dynamic changes of TNF-α、IL-1β and ICAM-1 in rats with severe acute pancreatitis (SAP),and to assess the effects of pretreatment with mycophenolate mofetil (MMF) on them.Methods A total of 104 male Wistar rats were randomly divided into normal group,sham operation group,SAP group and MMF group.(1) In normal group,rats had just an incision on abdomen wall and sutured (n =8) ; (2) In sham operation group,rats underwent laparotomy with flipping over the pancreas and gendy striking five times by using blunt instrument and abdomen was closed (n =32); (3) In SAP group,SAP in rats was induced by the retrograde injection of sodium taurocholate (STC) into the pancreatic duct (n =32) ; (4) In MMF group,MMF (250mg/kg)was injected intraperitoneally one hours after injection of STC (n =32).Rats were saerificed by abdominalaorta exsanguination separately at intervals of 3,6,12 and 24 h after modeling of pancreatitis.Histologic score of pancreatic tissue was assessed,and serum amylase (AMS) and serum C-reactive protein (CRP)were measured by automatic biochemical detector.The serum levels of inflammatory cyctokines (TNF-α,1L-1β and 1CAM-1 ) were measured by using ELISA,and expression of NF-κB mRNA in pancreatic tissue was measured by using quantitative PCR.Results Compared with sham operation group,histologic score of pancreatic tissue,serum amylase,TNF-α、IL-1β and ICAM-1 in SAP group were increased significantly (P ＜ 0.05 ).Pretreatment with MMF obviously deceased histologic score,and the serum levels of amylase,TNF-α,IL-1β and ICAM-1 and the expression of NF-κB mRNA were higher than those in SAP group in early stage (P ＜0.05 ).Conclusions Cytokines play an important role in early stage of SAP.MMF could attenuate the severity of inflammatory response through inhibiting cellular immunity and modulating the proinflammatory cytokines produced.

A rapid, sensitive and reliable diagnostic test for scrub typhus in China.

Purpose: To evaluate the performances for detection of IgM and IgG antibodies to Orientia. tsutsugamushi (Ot) using a gold conjugate-based rapid diagnostic test (RDT). Materials and Methods: The RDT employing mixture recombinant 56-kDa proteins of O. tsutsugamushi and the mIFA assay was performed on 33 patients from Fujian and Yunnan province respectively and 94 positive sera (36 from Hainan province and 58 from Jiangsu province) from convalescent stages of the patients with scrub typhus respectively and 82 negative sera from healthy farmers from Anhui province and Beijing City respectively in 2009. A comparison of the RDT and mIFA assay was performed by using the c2 test and the P level of ≤0.05 was considered to be significant. Results: Among these 94 positive sera from convalescent stages of the illness and 82 sera from control farmers, the specificity of RDT was 100% for both IgM and IgG tests. In 33 cases with scrub typhus, 5 cases were positively detected earlier by RDT than by mIFA for the IgM test, and 2 cases were positive for the IgG test. The sensitivities of RDT were 93.9% and 90.9% for IgM and IgG, respectively. Considering IgM and IgG together, the sensitivity was 100%. The geometric mean titre (GMT) of IFA and the RDT assay in diluted sera from confirmed cases were 1:37 versus 1:113 respectively (P<0.001) for IgM test and 1:99 versus 1:279 respectively (P<0.016) for IgG. Conclusions: The RDT was more sensitive than the traditional IFA for the early diagnosis of scrub typhus and was particularly suitable for use in rural areas.

Comparison of a rapid diagnostic test and microimmunofluorescence assay for detecting antibody to Orientia tsutsugamushi in scrub typhus patients in China

OBJECTIVE@#To evaluate the detection of IgM and IgG antibodies to Orientia tsutsugamushi (O. tsutsugamushi) by rapid diagnostic test (RDT) and microimmunofluorescence assay (mIFA).@*METHODS@#RDT using a mixture of recombinant 56-kDa proteins of O. tsutsugamushi and mIFA assay were performed on 20 patients from Fujian and 13 patients from Yunnan Province, and 82 sera samples from healthy farmers in Anhui Province and Beijing City in 2009. Comparison of the RDT and mIFA assay was performed by using X(2) test and the P level of <0.05 was considered to be significance.@*RESULTS@#Among these 82 normal sera samples, the specificity of RDT was 100% for both IgM and IgG tests. In 33 samples from patients with scrub typhus, 5 cases were positively detected earlier by RDT than by mIFA in IgM test, and 2 cases were positive in IgG test. Sensitivities of RDT were 93.9% and 90.9% for IgM and IgG, respectively. The sensitivity of combination test of IgM and IgG was 100%. Geometric mean titer diluted sera from confirmed cases by IFA and RDT assay were 1:37 vs. 1:113 (P<0.001) in IgM test and 1:99 vs. 1:279 (P<0.05) in IgG test.@*CONCLUSIONS@#RDT is more sensitivite than mIFA in the early diagnosis of scrub typhus and it is particularly applicable in rural areas.

Investigation of anaplasmosis in Yiyuan County, Shandong Province, China

OBJECTIVE@#To investigate the situation of anaplasmosis in Yiyuan county, Shandong Province.@*METHODS@#A total of 26 blood samples from febrile patients suspected of anaplasmosis, 48 blood samples from healthy farmers, 8 from dogs, and 10 from goats and 170 ticks were collected in the same area during 2005-2007, and detected by serological and molecular methods.@*RESULTS@#Eight confirmed cases and 6 probable cases were determined using serologic and molecular methods. The seroprevalence of Anaplasma phagocytophilum (A. phagocytophilum) was 26.7% in healthy cases. Nine out of 10 sheep samples and 7 out of 8 dog samples reacted positively to the A. phagocytophilum antigen. PCR amplification and sequencing of the 16SrRNA of A. phagocytophilum gene showed that some samples from patients, goats and ticks were 100% identical. The seroprevalence of Rickettsia typhi was 22.9%, Orientia tsutsugamushi 6.3%, Rickettsia sibirica 27.1%, Coxiella burnetii 18.8%, Bartonella henselae 31.3%, and Borrelia burgdorferi 41.6%.@*CONCLUSIONS@#It is important to make differential diagnosis of febrile patients and to apply treatment with specific antibiotics. It is needed to enforce essential prevention and control measures including tick control and to improve sanitation conditions.