1.Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing
Li Ya ZHANG ; Zhe Wen SU ; Chen Rui WANG ; Yan LI ; Feng Jun ZHANG ; Hui Sheng LIU ; He Dan HU ; Xiao Chong XU ; Yu Jia YIN ; Kai Qi YIN ; Ying HE ; Fan LI ; Hong Shi FU ; Kai NIE ; Dong Guo LIANG ; Yong TAO ; Tao Song XU ; Feng Chao MA ; Yu Huan WANG
Biomedical and Environmental Sciences 2024;37(3):294-302
Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
2.Gene Mutation Types of Thalassemia in Chongzuo Childbearing-age Population of Guangxi Zhuang Autonomous Region of China.
Dong-Ming LI ; Xiu-Ning HUANG ; Huan ZHAO ; Xiang CHEN ; Wan-Wei YANG ; Zhen-Ren PENG ; Li-Fang LIANG ; Bi-Yan CHEN ; Sheng HE
Journal of Experimental Hematology 2023;31(6):1804-1810
OBJECTIVE:
To investigate the gene mutation and genotype distribution of thalassemia in the population of childbearing age in Chongzuo area of Guangxi.
METHODS:
Six α-thalassemia and 17 β-thalassemia gene mutations common in Chinese were detected by gap-polymerase chain reaction (gap-PCR) combined with agarose gel eletrophoresis and reserve dot bolt hybridization in 29 266 cases of child-bearing age suspected of thalassemia.
RESULTS:
A total of 19 128 (65.36%) cases were identified with thalassemia. The detection rate of α-thalassemia, β-thalassemia and α-combining β-thalassemia was 45.25% (13 242/29 266), 15.47% (4 526/29 266) and 4.65% (1 360/29 266), respectively. A total carrying rate of 8 kinds of α-thalassemia gene mutations was 26.74% (15 649/58 532), including 12.51% for --SEA, followed by 5.70% for -α3.7, and 0.24% for --Thai. Among 32 α-thalassemia genotypes, the most common five were --SEA/αα, -α3.7/αα, αCSα/αα, -α4.2/αα and αWSα/αα, accounting for 47.27%, 18.31%, 8.56%, 8.52% and 7.91%, respectively, as well as 0.97% for --Thai/αα. A total carrying rate of 13 kinds of β-thalassemia gene mutations was 10.07% (5 897/58 532), including 3.63% for CD41-42, followed by 2.55% for CD17, and 0.003% for -50 (G>A). Among 17 β-thalassemia genotypes, the most common six were CD41-42/N, CD17/N, CD71-72/N, CD26/N, 28/N and IVSI-1/N, accounting for 36.15%, 25.81%, 9.43%, 8.18%, 8.09% and 7.75%. The homozygous genotype CD26/CD26 [hemoglobin (Hb): 121 g/L] and -28/-28 (Hb: 56 g/L) were respectively detected in one case, and double heterozygous genotype were detected in 5 cases, including 3 cases of CD41-42/CD26 (Hb: 41 g/L, 51 g/L, 63 g/L, respectively), 1 case of -28/IVSI-1 (Hb: 53 g/L), and 1 case of CD71-72/CD26 (Hb: 89 g/L), in which patients with moderate or severe anemia had a history of blood transfusion. Among 104 α-combining β-thalassemia genotypes, the most common were --SEA/αα, -α3.7/αα combining CD41-42/N and --SEA/αα combining CD17/N, accounting for 12.13%, 9.63% and 9.26%, respectively. In addition, 1 case of --SEA/-α3.7 combining -28/IVSI-1 (Hb: 83 g/L) and 1 case of -α3.7/αα combining CD41-42/ CD41-42 (Hb: 110 g/L) were detected without history of blood transfusion, while 1 case of αWSα/αα combining CD41-42/CD17 (Hb: 79 g/L) and 1 case of --SEA/αα combining CD17/-28 (Hb: 46 g/L) were detected with history.
CONCLUSIONS
The detection rate of thalassemia genes is high and the mutations are diverse in the population of childbearing age in Chongzuo area of Guangxi. The common deletion genotype is --SEA/αα in α-thalassemia and CD41-42/N in β-thalassemia, and deletion genotype --Thai is not rare. There is a certain incidence of intermediate and severe β-thalassemia, and most patients require transfusion therapy. The results are beneficial for genetic consultation and intervention of thalassemia.
Humans
;
beta-Thalassemia/genetics*
;
alpha-Thalassemia/genetics*
;
Dipeptidyl Peptidase 4/genetics*
;
China/epidemiology*
;
Genotype
;
Mutation
3.Clinical features and FGFR3 mutations of children with achondroplasia.
Hui-Qin ZHANG ; Dong-Ying TAO ; Jing-Jing ZHANG ; Huan-Hong NIU ; Jian-Feng LUO ; Sheng-Quan CHENG
Chinese Journal of Contemporary Pediatrics 2022;24(4):405-410
OBJECTIVES:
To study the clinical features and fibroblast growth factor receptor 3 (FGFR3) gene mutations of children with achondroplasia (ACH) through an analysis of 17 cases.
METHODS:
A retrospective analysis was performed on the clinical data and FGFR3 gene detection results of 17 children with ACH who were diagnosed from January 2009 to October 2021.
RESULTS:
Of the 17 children with ACH, common clinical manifestations included disproportionate short stature (100%, 17/17), macrocephaly (100%, 17/17), trident hand (82%, 14/17), and genu varum (88%, 15/17). The common imaging findings were rhizomelic shortening of the long bones (100%, 17/17) and narrowing of the lumbar intervertebral space (88%, 15/17). Major complications included skeletal dysplasia (100%, 17/17), middle ear dysfunction (82%, 14/17), motor/language developmental delay (88%, 15/17), chronic pain (59%, 10/17), sleep apnea (53%, 9/17), obesity (41%, 7/17), foramen magnum stenosis (35%, 6/17), and hydrocephalus (24%, 4/17). All 17 children (100%) had FGFR3 mutations, among whom 13 had c.1138G>A hotspot mutations of the FGFR3 gene, 2 had c.1138G>C mutations of the FGFR3 gene, and 2 had unreported mutations, with c.1252C>T mutations of the FGFR3 gene in one child and c.445+2_445+5delTAGG mutations of the FGFR3 gene in the other child.
CONCLUSIONS
This study identifies the unreported mutation sites of the FGFR3 gene, which extends the gene mutation spectrum of ACH. ACH is a progressive disease requiring lifelong management through multidisciplinary collaboration.
Achondroplasia/genetics*
;
Child
;
Humans
;
Mutation
;
Osteochondrodysplasias/genetics*
;
Receptor, Fibroblast Growth Factor, Type 3/genetics*
;
Retrospective Studies
4.Genomic and transcriptomic analysis unveils population evolution and development of pesticide resistance in fall armyworm Spodoptera frugiperda.
Furong GUI ; Tianming LAN ; Yue ZHAO ; Wei GUO ; Yang DONG ; Dongming FANG ; Huan LIU ; Haimeng LI ; Hongli WANG ; Ruoshi HAO ; Xiaofang CHENG ; Yahong LI ; Pengcheng YANG ; Sunil Kumar SAHU ; Yaping CHEN ; Le CHENG ; Shuqi HE ; Ping LIU ; Guangyi FAN ; Haorong LU ; Guohai HU ; Wei DONG ; Bin CHEN ; Yuan JIANG ; Yongwei ZHANG ; Hanhong XU ; Fei LIN ; Bernard SLIPPERS ; Alisa POSTMA ; Matthew JACKSON ; Birhan Addisie ABATE ; Kassahun TESFAYE ; Aschalew Lemma DEMIE ; Meseret Destaw BAYELEYGNE ; Dawit Tesfaye DEGEFU ; Feng CHEN ; Paul K KURIA ; Zachary M KINYUA ; Tong-Xian LIU ; Huanming YANG ; Fangneng HUANG ; Xin LIU ; Jun SHENG ; Le KANG
Protein & Cell 2022;13(7):513-531
The fall armyworm (FAW), Spodoptera frugiperda, is a destructive pest native to America and has recently become an invasive insect pest in China. Because of its rapid spread and great risks in China, understanding of FAW genetic background and pesticide resistance is urgent and essential to develop effective management strategies. Here, we assembled a chromosome-level genome of a male FAW (SFynMstLFR) and compared re-sequencing results of the populations from America, Africa, and China. Strain identification of 163 individuals collected from America, Africa and China showed that both C and R strains were found in the American populations, while only C strain was found in the Chinese and African populations. Moreover, population genomics analysis showed that populations from Africa and China have close relationship with significantly genetic differentiation from American populations. Taken together, FAWs invaded into China were most likely originated from Africa. Comparative genomics analysis displayed that the cytochrome p450 gene family is extremely expanded to 425 members in FAW, of which 283 genes are specific to FAW. Treatments of Chinese populations with twenty-three pesticides showed the variant patterns of transcriptome profiles, and several detoxification genes such as AOX, UGT and GST specially responded to the pesticides. These findings will be useful in developing effective strategies for management of FAW in China and other invaded areas.
Animals
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China
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Genomics
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Humans
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Male
;
Pesticides
;
Spodoptera/genetics*
;
Transcriptome
5.A new dimeric dehydrocorydaline alkaloid from vinegar-prepared Corydalis yanhusuo
Dong-jie FANG ; Gui-yang XIA ; Ling-yan WANG ; Huan XIA ; Yan-bin SUN ; Sheng LIN
Acta Pharmaceutica Sinica 2021;56(4):1096-1099
Bidehydrocorydaline A (
6.Biotransformation of α-asarone by Alternaria longipes CGMCC 3.2875.
Jian ZOU ; Shuai ZHANG ; Huan ZHAO ; Yong-Heng WANG ; Zheng-Qun ZHOU ; Guo-Dong CHEN ; Dan HU ; Ning LI ; Xin-Sheng YAO ; Hao GAO
Chinese Journal of Natural Medicines (English Ed.) 2021;19(9):700-705
Biotransformation of α-asarone by Alternaria longipes CGMCC 3.2875 yielded two pairs of new neolignans, (+) (7S, 8S, 7'S, 8'R) iso-magnosalicin (1a)/(-) (7R, 8R, 7'R, 8'S) iso-magnosalicin (1b) and (+) (7R, 8R, 7'S, 8'R) magnosalicin (2a)/(-) (7S, 8S, 7'R, 8'S) magnosalicin (2b), and four known metabolites, (±) acoraminol A (3), (±) acoraminol B (4), asaraldehyde (5), and 2, 4, 5-trimethoxybenzoic acid (6). Their structures, including absolute configurations, were determined by extensive analysis of NMR spectra, X-ray crystallography, and quantum chemical ECD calculations. The cytotoxic activity and Aβ
7.A new dimeric benzylisoquinoline alkaloid from Corydalis yanhusuo
Gui-yang XIA ; Biao-bing XIAO ; Ling-yan WANG ; Dong-jie FANG ; Huan XIA ; Sheng LIN
Acta Pharmaceutica Sinica 2021;56(2):553-556
(±)-Bicoryanhunine B (
8. Analysis on protein expression of ferritin light chain in alveolar macrophages of patients with occupational silicosis
Jie-jie QIAO ; Lu ZHANG ; Ke HAN ; Huan WANG ; Sheng-qiang DU ; Dong-xu REN ; Yu-qing JIN ; Xin XU ; Fu-hai SHEN
China Occupational Medicine 2021;48(05):519-522
OBJECTIVE: To observe the protein expression of ferritin light chain(FTL) in alveolar macrophages(AM) of patients with occupational silicosis(hereinafter referred to as silicosis). METHODS: The male patients with silicosis at stage Ⅰ, Ⅱ, and Ⅲ were separately selected as the silicosis groupⅠ, Ⅱ, and Ⅲ using judgment sampling method, with 15 patients in each group. Meanwhile, 15 male silicon dust workers with small lung shadows but not diagnosed as silicosis were selected as the control group. Bronchoalveolar lavage fluid was collected from the four groups, and AM was separated and purified, and protein was extracted after lysis of the AM. Western blotting was used to detect the relative expression of FTL protein in the AM. RESULTS: The relative expression of FTL protein in AM of silicosis groupⅠ, Ⅱ, and Ⅲ was lower than that in the control group(all P<0.05). The relative expression of FTL protein in AM decreased with the increase of silicosis stage(P<0.05).CONCLUSION: The expression of FTL protein in AM was down-regulated in patients with silicosis in a dose-response manner. It is speculated that FTL may have a negative regulatory effect in the progress of silicosis fibrosis.
9.Epidemiological analysis of novel coronavirus pneumonia in Huangpu District of Shanghai
Yu-liang HUANG ; Fei SU ; Ying-jun HAN ; Jia-ying LI ; Sheng-ying DU ; Yu-ming MAO ; Huan-zhu ZHANG ; Zhen-dong ZHANG ; Jing-xiong HE ; Xiao LIU ; Jing-xin ZHOU ; Qiang GAO ; Ling YAN ; Huai-xia YANG ; Yi-jun WANG ; Min SHU ; Fu-jie SHEN ; Lu LU
Shanghai Journal of Preventive Medicine 2020;32(9):726-
Objective To understand the epidemiological characteristics of COVID-19 epidemic in Huangpu District of Shanghai, and to provide scientific evidence for prevention and control of COVID-19. Methods Descriptive statistics were used to study the suspected and confirmed cases of COVID-19 reported from January 21 through March 10, 2020 in Huangpu District, Shanghai. Results A total of 120 suspected cases of COVID-19 were reported, of which 12 were diagnosed and 108 were excluded.The first confirmed case was reported on January 21, and the last case was on February 10; the majority (11/12) of the confirmed cases were reported from January 21 through February 1.The average duration of time from the symptom onset to the first medical visit was 2.6 days, whereas the average duration from the first medical visit to the hospital diagnosis was 2.2 days.There were 15 suspected cases with a confirmed history of residence or tourism in Wuhan, in which 6 were confirmed cases.Moreover, 5 suspected cases had a confirmed history of contact with other confirmed cases, in which 3 were confirmed cases.Thus, exposure in Wuhan and exposure to confirmed cases were the most significant risk factors at this stage of the epidemic. Conclusion The 12 cases identified in Huangpu District of Shanghai are all adults, half of whom had confirmed history of exposure in Wuhan.The first cluster of COVID-19 cases in Shanghai is documented in Huangpu District.Epidemiological investigation reveals that the confirmed cases might be infectious the day before the symptom onset.
10.Comparison of epidemiological and clinical characteristics of COVID-19 patients with and without Wuhan exposure history in Zhejiang Province, China.
Jiang-Shan LIAN ; Huan CAI ; Shao-Rui HAO ; Xi JIN ; Xiao-Li ZHANG ; Lin ZHENG ; Hong-Yu JIA ; Jian-Hua HU ; Shan-Yan ZHANG ; Guo-Dong YU ; Jue-Qing GU ; Chan-Yuan YE ; Ci-Liang JIN ; Ying-Feng LU ; Ji-Fang SHENG ; Yi-da YANG
Journal of Zhejiang University. Science. B 2020;21(5):369-377
BACKGROUND:
A novel coronavirus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), first identified in Wuhan, China, has been rapidly spreading around the world. This study investigates the epidemiological and clinical characteristics of coronavirus disease 2019 (COVID-19) patients in Zhejiang Province who did or did not have a history of Wuhan exposure.
METHODS:
We collected data from medical records of confirmed COVID-19 patients in Zhejiang Province from Jan. 17 to Feb. 7, 2020 and analyzed epidemiological, clinical, and treatment data of those with and without recorded recent exposure in Wuhan.
RESULTS:
Patients in the control group were older than those in the exposure group ((48.19±16.13) years vs. (43.47±13.12) years, P<0.001), and more were over 65 years old (15.95% control vs. 5.60% exposure, P<0.001). The rate of clustered onset was also significantly higher in the control group than in the exposure group (31.39% vs. 18.66%, P<0.001). The symptom of a sore throat in patients in the exposure group was significantly higher than that in the control group (17.30% vs. 10.89%, P=0.01); however, headache in the exposure group was significantly lower than that in the control group (6.87% vs. 12.15%, P=0.015). More patients in the exposure group had a significantly lower level of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) than those in the control group. There was no significant difference in any degree of COVID-19 including mild, severe, and critical between the two groups.
CONCLUSIONS
From the perspective of epidemiological and clinical characteristics, there was no significant difference between COVID-19 patients with and without Wuhan exposure history.
Adolescent
;
Adult
;
Aged
;
Aspartate Aminotransferases
;
blood
;
Betacoronavirus
;
Case-Control Studies
;
Child
;
Child, Preschool
;
China
;
epidemiology
;
Coronavirus Infections
;
epidemiology
;
physiopathology
;
therapy
;
Female
;
Humans
;
Infant
;
Infant, Newborn
;
L-Lactate Dehydrogenase
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blood
;
Male
;
Middle Aged
;
Pandemics
;
Pneumonia, Viral
;
epidemiology
;
physiopathology
;
therapy
;
Retrospective Studies
;
Young Adult

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