1.High-level expression and purification of human creatine kinase MM isozymes in E.coli
Yan HUANG ; Wenshuang YANG ; Guobin XU ; Tiean XIA
Chinese Journal of Clinical Laboratory Science 1985;0(04):-
Objective To construct a prokaryotic expression system for human creatine kinase(CK) MM isozyme,purify the recombinant protein of CK expressed in Eschericheia coli(E.coli) and examine the stability of the recombinant protein for its application in CK measurement system as the quality control.Methods Total RNA of extracted from fetal cardiac muscle was reversetranscripted and cDNA encoding human CK was amplified which was inserted into pET-15b plasmid vector.The recombinant plasmid was transfered into E.coli BL21(DE3) and induced by isopropyl-?-D-thiogalactopyranoside(IPTG).Recombinant CK-MM was separated from bacterial proteins by affinity chromatography on a Ni2+-Sepharose column.The activity of the recombinant enzyme was observed in different matrixes.Results The enzymatic activity of the crude extracts of CK-MM was up to 280,000U/L.After one step affinity chromatography,the fusion protein showed a single band in SDS-PAGE gel.The purified protein was stable and the enzymatic activity remained unchanged for a month in the matrix containing bovine serum albumin,EDTA and 1,4-Dithiothreitol(DTT).Conclusion The recombinant CK-MM showed key properties of the native creatine kinase isozyme and may be used as control and calibrator for determination of serum CK-MM.
2.Analysis of 2864 cases of pregnant women in a single center results of oral 75 g anhydrous glucose tolerance test
Le CHANG ; Yaping JIANG ; Shiju JIANG ; Hongyun YANG ; Haixia LI ; Xuejing WANG ; Wenshuang YANG ; Guobin XU
Chinese Journal of Laboratory Medicine 2012;35(6):517-520
Objective Women with gestational diabetes mellitus (GDM) are at risk to develop Type 2 diabetes mellitus ( DM2).The aim of this study is to compare the effect of three diagnostic criteria on the prevalence of GDM.Methods 2864 pregnant women receiving obstetric care at 24 to 28 weeks of gestation at Peking University First Hospital,underwent the 75 g oral glucose tolerance test for routine GDM screening.The pregnancies were evaluated bythe American Diabetes Association (ADA 2004) Cuideline,the new criteria which was proposed by the International Association of Diabetes and Pregnancy Study Groups (IADPSG 2010) based on the HAPO study,not by the criteria of American College of Obstetricians and Chinese Obstetrics and Gynecology (6th edition 2005).Venous plasma glucose was measured by the IDMS-traceable glucose oxidase method.Results The mean age of the women was 30 years.590 women were diagnosed with GDM using the IADPSG criteria.According to the IADPSG standard,GDM diagnostic rate reached 20.6%.The diagnostic rate of new criteria was increased by 307% and 199% compared with the Chinese Obstetrics and Gynaecology (6th edition) and the old ADA standard,respectively.If increasing the number of abnormal glucose values in the IADPS standard to 2 or more,the number of positive cases dropped to 227 people.The diagnostic rate of GDM plummeted to 7.93%,similar with that of the old ADA diagnostic criteria (6.8%).Conclusions The diagnosis rate of GDM was significantly higher using the IADPSG criteria,due to more stringent glucose abnormality recommended by the IADPSG.
3.Value of EAT-10 in screening dysphagia:a meta-analysis
Wenshuang YANG ; Houqiang HUANG ; Silin ZHENG
Modern Clinical Nursing 2017;16(8):25-30
Objective To evaluate the diagnostic value of the Eating Assessment Tool (EAT-10) in screening dysphagia by using 2 and 3 as cut-off value with systematic review. Methods A retrieval on the diagnostic value of EAT-10 in dysphagia was done all through PubMed, Embase, the Cochrane Library, CBM, CNKI, WANFANG and VIP databases (from January 2008 to November 2016)."Quality As-sessment of Diagnostic Accuracy Studies" (QUADAS) was used to appraise included papers. Meta-Disc1.4 software was used to analyze heterogeneity and the combined effect of the data. Results Three papers were included and 394 subjects were included in the review. By using 2 and 3 as the cut-off value of EAT-10, the value of the weighted sensitivity was 0.84 [95%CI (0.79, 0.88)] and 0.77 [95%CI(0.71, 0.82)], the specificity was 0.64[95%CI (0.55, 0.72)] and 0.74 [95%CI(0.66, 0.81)],and the area under the receiver-operating-characteris-tic-curve (AUC-ROC) was 0.8621 and 0.8343, which illustrated the diagnostic accuracy of EAT-10 on dysphagia at the medium level. Conclusion The diagnostic ability of EAT-10 is good, but the sensitivity and specificity are different by using 2 and 3 as the cut-off value. The subjects should consider the specific clinical situation to determine the best cut-off value.
4.The application values of MR quantitative susceptibility mapping in differentiation between knee meniscus degeneration and tear
Ye LI ; Aocai YANG ; Tao GONG ; Sai SHAO ; Xianyun CAI ; Cong SUN ; Tuantuan WANG ; Wenshuang ZHANG ; Weibo CHEN ; Guangbin WANG
Chinese Journal of Radiology 2021;55(5):522-527
Objective:To evaluate the feasibility and the application values of quantitative susceptibility mapping (QSM) for the assessment of meniscal injury and in distinguishing meniscus degeneration and tears.Methods:The clinical and imaging data of 70 patients suspected of meniscus injury and scheduled for arthroscopy in Shandong Medical Imaging Research Institute, Cheeloo College of Medicine, Shandong University from November 2019 to June 2020 were analyzed retrospectively. Thirty age-and sex-matched healthy subjects were also examined as controls. All subjects received knee joint QSM and routine MR imaging. According to the results of arthroscopy, the patients was divided into meniscus degeneration and meniscus tear groups, respectively. The conventional MR was evaluated by two radiologists. The meniscus injury area was delineated on the original QSM magnitude images (the central area of the posterior corner of the lateral meniscus was selected in the healthy controls) and mapped to the corresponding QSM maps, and the magnetic susceptibility values were measured. Kruskal-Wallis H test was used to analyze the magnetic sensitivity values of meniscal degeneration, meniscal tear and healthy control groups; and Bonferroni was used to correct the pairwise comparison. ROC curve was established to evaluate the threshold and efficacy of magnetic susceptibility value in the diagnosis of meniscal tear. The results were compared with those of conventional MRI. Results:The magnetic susceptibility values of meniscus of healthy controls, meniscal degeneration and meniscal tear groups were (0.035±0.016)ppm, -0.031(-0.040,-0.005)ppm, and(-0.122±0.115)ppm, respectively, with significant difference found among the three groups (χ2=44.419, P<0.05). The magnetic susceptibility values of meniscus of healthy controls was significantly higher than those of meniscus degeneration patients and meniscus tear patients (χ2=-23.843, -48.253, P<0.05). The magnetic susceptibility values of meniscus of meniscus tear group was significant lower than those of meniscus degeneration group (χ2=-24.410, P<0.05). Taking magnetic susceptibility values of -0.062 5 ppm as threshold, the area under the ROC curve for the diagnosis of meniscal tears was 0.949, with the sensitivity as 87% and the specificity as 100%. The sensitivity and specificity of conventional MRI in the diagnosis of meniscal tears were 86.8% and 87.5%, respectively. Conclusion:QSM can quantitatively evaluate meniscus injury and can be used as an effective supplement method to conventional MRI, which is helpful to improve the diagnosis of meniscus tear.
5.Plasmablastic lymphoma: a clinicopathologic analysis of 11 cases with review of literature.
Fen LI ; Wenshuang DING ; Zhuo ZUO ; Ning GENG ; Huai YANG ; Xiulan LIU ; Jianchao WANG ; Wenqing YAO ; Weiping LIU
Chinese Journal of Pathology 2016;45(1):37-42
OBJECTIVETo investigate the clinicpathologic features and diagnosis of plasmablastic lymphoma (PBL).
METHODSEleven cases of PBL were collected and followed up, with review of the literature. HIV and EBV status and their relationships with the tumor were specially compared as well.
RESULTSIn the current cohort, 10 patients were serologically HIV negative; the male to female ratio was 8 to 3, and the median age was 57 years. Ten cases showed extranodal involvement and one case was nodal based. At presentation, five patients had mid-facial involvement, including sinonasal area (3 cases) and oral cavity (2 cases). Histologically, six were PBL of oral mucosa type, and five were PBL with plasmacytic differentiation. In all cases, the neoplastic cells expressed CD138 and MUM-1, and were negative for CD20 and CD3ε; the median Ki-67 index was 80%. Five cases were EBER1/2 in situ hybridization positive. IgH or/and Igκ gene rearrangement was detected in all five cases examined.
CONCLUSIONSMost patients were no congenital or acquired immunodeficiency in the retrospective study. Of the died patients, EBER1/2 in situ hybridization were negative and their disease staging were Ⅳ, The neoplastic cells were immunoblastic or plasmablastic, sometimes the plasmacytoid cell can be seen and the neoplastic cell had mature plasma cell phenotype, the pathologic diagnosis of the lymphoma is still controversial now. Differentiate with plasma cell neoplasm is difficult, it is necessary to accumulate more cases for advanced study and observation in the future.
Female ; Gene Rearrangement ; Humans ; In Situ Hybridization ; Male ; Middle Aged ; Multiple Myeloma ; Plasma Cells ; Plasmablastic Lymphoma ; diagnosis ; pathology ; RNA, Viral ; metabolism ; Retrospective Studies
6.The application of three statistical methods for reference intervals establishment of coagulation assays during pregnancy
Jingchen ZHAO ; Wenshuang YANG ; Shiju JIANG ; Ying XING ; Linzi MIAO ; Chenxue QU
Chinese Journal of Laboratory Medicine 2020;43(8):823-830
Objective:To compare the practical value of three statistical methods in establishing reference intervals by the indirect method.Methods:This is a methodological evaluation study. The data of coagulation parameters were obtained from laboratory information system, which were from pregnant women who had done prenatal examination in Peking University First Hospital from January 2017 to December 2019. The test results from 32 401 pregnant women were collected. Those healthy pregnant women were divided into three groups: early pregnancy group(n=11 151), middle pregnancy group(n=4 872) and late pregnancy group(n=16 378). Statistical analysis was performed for the result of PT, APTT, FIB, TT, D-D and FDP, and the necessity of stratification based on age in different periods of pregnancy was analyzed. Stratification based on age was necessary in three pregnancy groups for PT and APTT, in late pregnancy group for FIB and TT, and in early pregnancy group for D-D. The reference intervals of coagulation parameters were calculated by three statistical methods: non-parametric method, Hoffmann method and Q-Q plot method. Forty-two healthy pregnant women from October 2019 to January 2020 were enrolled as reference individuals for the validation of the reference intervals.The proportions of test results outside the reference intervals in the reference population calculated and compared.Results:The levels of the six coagulation assays vary significantly during the three periods of pregnancy, stratification based on age was necessary in three pregnancy groups for PT and APTT, in late pregnancy group for FIB and TT, and in early pregnancy group for D-D. If the number of test results was large, non-parametric and Hoffmann method provided more similar results, while the reference intervals calculated with Q-Q plot method was slightly wider than Hoffmann method. If the number of test results was small, reference intervals calculated with Hoffmann and Q-Q plot method were more reliable. For pregnant women during early pregnancy under the age of 35, the reference intervals of PT, APTT, FIB and TT calculated by this method were (10.44-13.11)s, (25.29-35.88)s, (2.61-4.64)g/L and (11.53-15.58)s.Conclusion:When establishing the reference interval, stratification according to pregnancy period and age was needed. Hoffmann method can be used as an alternative to the direct method.
7.A rare case of dicentric ring chromosome and derivative ring chromosome Chimera.
Junzhen ZHU ; Xiaoping YU ; Xiaofeng QI ; Qinying CAO ; Wenshuang ZHU ; Dan YANG ; Haoyu ZHANG ; Zhanyun SONG ; Shibo WANG ; Cuixia WANG
Chinese Journal of Medical Genetics 2022;39(5):534-536
OBJECTIVE:
Utilize high-resolution chromosome analysis and microarray detection to determine the genetic etiology of infertility of a 32-year old female patient.
METHODS:
The peripheral blood of the patient was cultured for high-resolution chromosome G and C banding karyotype analysis, and then 750K SNP-Array chip detection was performed.
RESULTS:
Karyotype analysis results showed that the patient's karyotype was 45,XX,-13 [7]/46,XX,r(13) (p13q34) [185]/46,XX,dic r(13;13)(p13q34;p13q34) [14]/ 47,XX,+der(13;13;13;13) (p13q34;p13q34;p13q34; p13q34), dic r(13;13) [1]/ 46,XX [3]. The microarray results showed that the patient had a 3.3 Mb deletion in the 13q34 segment of chromosome 13, which may be related to infertility.
CONCLUSION
Infertility of the patient reported in this article may be related to the deletion of chromosome segment (13q34-qter).
Adult
;
Chimera
;
Chromosome Banding
;
Chromosome Deletion
;
Chromosome Disorders/genetics*
;
Dacarbazine
;
Female
;
Humans
;
Infertility/genetics*
;
Ring Chromosomes
8.The value of MRI anterior cartilaginous acetabulum-head-index to evaluate hip function after treatment of developmental dysplasia of the hip
Wenshuang ZHANG ; Yanzhou WANG ; Tianyou LI ; Cong SUN ; Qinhua LUAN ; Yongguang BAN ; Yufan CHEN ; Aocai YANG ; Ye LI ; Guangbin WANG
Chinese Journal of Radiology 2021;55(10):1076-1081
Objective:To evaluate the feasibility of high resolution MRI for the measurement of anterior cartilaginous acetabulum-head-index (A-CAHI) and the value of A-CAHI for predicting hip clinical function after treatment in developmental dysplasia of the hip (DDH).Methods:The imaging data of 92 hips from 61 children with treated DDH were retrospectively reviewed in Shandong Medical Imaging Research Institute from January 2019 to January 2020. All children underwent conservative treatments or surgical interventions 3 years ago. Hip function after treatment was evaluated clinically based on the modified MacKay criteria. The hips were divided into satisfactory clinical function group (McKay rating excellent or good, n=46) and unsatisfactory group (McKay rating fair or poor, n=46). All patients were imaged with conventional MRI, high resolution fat suppressed proton density weighted image (FS-PDWI) of the unilateral hip joint in oblique sagittal view, and anteroposterior hip radiographs. A-CAHI and lateral cartilaginous acetabulum-head-index (L-CAHI) were measured respectively on high-resolution oblique sagittal PDWI and conventional coronal T 1WI. Acetabulum head index (AHI) was also measured on anteroposterior hip radiograph. Mann-Whitney U test or independent-samples t test was used to compare the difference of A-CAHI, L-CAHI and AHI between satisfactory and unsatisfactory clinical function groups. The diagnostic value using A-CAHI, L-CAHI, AHI, or A-CAHI combined with L-CAHI for unsatisfactory clinical function were investigated by the ROC curve. The area under the curve (AUC) and the Z statistic were used to compare diagnostic performance. Results:The values of A-CAHI, L-CAHI and AHI were significantly higher in satisfactory clinical function group compared with the unsatisfactory group ( Z=-7.746, -7.735, t=-7.199, all P<0.001).A-CAHI combined with L-CAHI had the significant highest diagnostic accuracy compared with A-CAHI, L-CAHI and AHI (AUC were 0.994, 0.969, 0.968, 0.861, respectively), with significant differences ( Z=1.975, 2.006, 3.553, P=0.048, 0.051,<0.001). The sensitivity and specificity of A-CAHI combined with L-CAHI for the diagnosis of prognosis were 95.7% and 97.8%, respectively. Conclusions:A-CAHI measured by high resolution MRI was found to have the highest diagnostic accuracy for prediction of hip clinical function in the treated DDH, and combined with L-CAHI can improve the diagnostic accuracy significantly.
9.Age-related study of brachial plexus MRI contrast to noise ratio and diffusion tensor imaging values in normal adults
Yufan CHEN ; Shanshan WANG ; Cong SUN ; Hong TANG ; Wenshuang ZHANG ; Aocai YANG ; Guangbin WANG
Chinese Journal of Radiology 2020;54(11):1095-1100
Objective:To investigate the correlation of age with diffusion tensor imaging (DTI) values as fractional anisotropy (FA), apparent diffusion coefficient (ADC) and contrast to noise ratio (CNR) of three-dimensional nerve-sheath signal increased with inked rest-tissue rapid acquisition of relaxation imaging (3D SHINKEI) of the brachial plexus in normal adults.Methods:A total of 54 adult healthy volunteers and 6 patients with Guillain-Barre syndrome were prospectively enrolled from October 2018 to April 2019. Healthy volunteers were divided into 3 groups according to age as 21-40 years old group ( n=20), 41-60 years old group ( n=20), and ≥61 years old group ( n=14). All of them underwent MRI examination of the brachial plexus, including DTI and 3D SHINKEI sequences. The average FA and ADC values of the brachial plexus were measured and calculated through the fusion of DTI and 3D SHINKEI by 2 physicians independently. The contrast to noise ratio (CNR) of brachial plexus nerve was measured in 3D SHINKEI sequence images. Intraclass correlation efficient (ICC) was used to analyze the consistency between the two physicians. A simple linear regression model and Pearson correlation analysis were used to detect the correlation between FA, ADC, CNR and age. One-way analysis of variance was used to compare the differences of FA, ADC and CNR in different age groups. The FA and ADC values in different genders were compared by independent sample t-test. Results:Inter-observer agreements of the 2 physicians were good for FA and ADC values with ICC values of 0.811 and 0.901, respectively. For different groups, FA values were 0.397±0.023, 0.368±0.023, and 0.334±0.018 and ADC values were (1.376±0.072) × 10 -3 mm 2/s, (1.466±0.086) × 10 -3 mm 2/s, (1.486±0.080) × 10 -3 mm 2/s, for 21-40, 41-60, and ≥61 years old groups, respectively with statistical significant difference ( F=25.311, P<0.001; F=9.948, P<0.001). The CNR of the brachial plexus were 510.583±192.846, 502.581±128.821, and 426.782±113.648 for 21-40, 41-60, and ≥61 years old group without statistical difference ( F=1.429, P=0.249). The FA value of brachial plexus was highly negatively correlated with age ( r=-0.745, P<0.001), while the ADC value was moderately positively correlated with age ( r=0.596, P<0.001). The CNR of 3D SHINKEI sequence was negatively correlated with age ( r=-0.292, P=0.033). There was no statistically significant difference in brachial plexus FA and ADC values between male and female subjects ( t=1.496, P=0.141; t=-1.557, P=0.126). The FA value of Guillain-Barre syndrome patients was lower than that of healthy volanteers in the same age group ( t=6.129, P<0.001), and the ADC value had no statistical diference ( t=-1.335, P=0.186). Conclusion:The values of FA, ADC and CNR of brachial plexus in normal adults change with age. Among them, FA value is more significant.
10.A multi-center survey on the application of antibacterial agents in Chinese children in 2019
Jiaosheng ZHANG ; Xiang MA ; Lanfang TANG ; Daiyin TIAN ; Li LIN ; Yanqi LI ; Jing QIAN ; Wenshuang ZHANG ; Wei LI ; Gen LU ; Ligang SI ; Ping JIN ; Liang ZHU ; Keye WU ; Jikui DENG ; Yuejie ZHENG ; Yonghong YANG
Chinese Journal of Applied Clinical Pediatrics 2022;37(14):1074-1081
Objective:To analyze the patterns of antibacterial agents in Chinese children surveyed by the China multi-center monitoring network for the application of antibacterial agents in children and neonates in 2019 by using World Health Organization (WHO) Access, Watch, Reserve and Not-recommended (AWaRe) and typical anatomical/therapeutic/chemical (ATC) in this study.Methods:The cross-sectional method was adopted.A multi-center cross-sectional survey was conducted on one day from September to December 2019.The information of all inpatients taking antibiotics was uploaded to the network-based data collection system (https: //garpec-31.mobilemd.cn/login.aspx? relogin=true). This study covered 13 hospitals from 10 provinces and cities in China.All hospitalized children in the Respiratory Department, Infectious Disease Department, General Surgery Department, Pediatric Intensive Care Units, Neonatal Intensive Care Units and Neonatology joined in this survey.The clinically used antibacterial agents were classified by AWaRe and ATC, and the AWaRe and ATC distributions of antibacterial agents prescribed for Chinese children and neonates were described.Results:Of the 2 644 antibiotic prescriptions included from 13 hospitals, 2 134 (80.71%) were for children and 510 (19.29%) were for neonates.Of all antibiotic prescriptions, there were 368 (13.92%) Access antibiotics prescriptions, 1 973 (74.62%) Watch prescriptions, 60 (2.27%) Reserve prescriptions and 243 (9.19%) Not-recommended prescriptions.The top-five antibiotics prescribed for children and neonates were third-generation cephalosporins (1 056, 39.94%), macrolides (492, 18.61%), carbapenems (275, 10.40%), beta lactam-beta lactamase inhibitors (246, 9.30%), and second-generation cephalosporins (136, 5.14%). The use ratios of Access, Watch, Reserve and Not-recommended antibiotics in each center ranged from 0 to 30.00%, 36.67% to 97.20%, 0 to 17.02% and 0 to 33.33%, respectively.In 1 360 antibiotic prescriptions for children and neonates with pneumonia, there were 152 (11.18%) Access antibiotics, 1 051 (77.28%) Watch antibiotics, 37 (2.72%) Reserve antibiotics, and 120 (8.82%) Not-recommended antibiotics.The top-five antibiotics prescribed for children with pneumonia were third-generation cephalosporins (522, 38.38%), macrolides (388, 28.53%), beta lactam-beta lactamase inhibitors (141, 10.37%), carbapenems (117, 8.6%) and penicillins (49, 3.60%).Conclusions:Watch antibiotics and broad spectrum antibiotics such as third-generation cephalosporins and macrolides prone to induce resistance are the main antibacterial agents used in Chinese children and neonates with pneumonia.Broad-spectrum antibiotics may be overused in Chinese children and neonates.