Objective To investigate the application value of single nucleotide polymorphism(SNP)linkage analysis based on next-generation sequencing(NGS)technology in preimplantation genetic testing(PGT)of families with autosomal recessive polycystic kidney disease(ARPKD).Methods A family with ARPKD was selected,where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus.Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1),c.10444C>T(paternal)and c.4303del(maternal),with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene,335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR)and NGS.The couple′s SNP risk haplotypes carrying gene mutations were constructed.After in vitro fertilization,blastocyst culture was performed.Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification,and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos.Sanger sequencing was used to verify the results of embryo linkage analysis.Results Among the 6 biopsied embryos,4 were mutation-free and euploid,1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data,making it impossible to judge.One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus,resulting in the full-term delivery of a healthy baby.Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families,while avoiding abortion issues caused by aneuploid embryos.This study is also the first PGT report target-ing the PKHD1 gene c.4303del mutation.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.

BACKGROUND: Maturity-onset diabetes of the young (MODY) is the most common monogenic diabetes. The aim of this study was to assess the prevalence of MODY in phenotypic type 2 diabetes (T2DM) among Chinese young adults. METHODS: From April 2015 to October 2017, this cross-sectional study involved 2429 consecutive patients from 46 hospitals in China, newly diagnosed between 15 years and 45 years, with T2DM phenotype and negative for standardized glutamic acid decarboxylase antibody at the core laboratory. Sequencing using a custom monogenic diabetes gene panel was performed, and variants of 14 MODY genes were interpreted as per current guidelines. RESULTS: The survey determined 18 patients having genetic variants causing MODY (6 HNF1A , 5 GCK , 3 HNF4A , 2 INS , 1 PDX1 , and 1 PAX4 ). The prevalence of MODY was 0.74% (95% confidence interval [CI]: 0.40-1.08%). The clinical characteristics of MODY patients were not specific, 72.2% (13/18) of them were diagnosed after 35 years, 47.1% (8/17) had metabolic syndrome, and only 38.9% (7/18) had a family history of diabetes. No significant difference in manifestations except for hemoglobin A1c levels was found between MODY and non-MODY patients. CONCLUSION The prevalence of MODY in young adults with phenotypic T2DM was 0.74%, among which HNF1A -, GCK -, and HNF4A -MODY were the most common subtypes. Clinical features played a limited role in the recognition of MODY.
Humans ; Diabetes Mellitus, Type 2/diagnosis* ; Cross-Sectional Studies ; Mutation ; Prevalence ; Phenotype

Objective To observe the effect of pre-hospital and intra-hospital collaborative ther-apeutic model in the treatment of patients with acute cerebral infarction.Methods A total of 67 pa-tients with acute cerebral infarction were selected as the research objects,and they were divided into observation group(n=37)and control group(n=30)according to the therapeutic model.The con-trol group was conducted with the traditional therapeutic model,while the observation group was con-ducted with the pre-hospital and intra-hospital collaborative therapeutic model.The time efficiency of intravenous thrombolysis,early recovery of nerve function and oxidative stress indexes were compared between the two groups.Results There was no significant difference in the time from onset to visit between the two groups(P＞0.05);the time from seeing a doctor to thrombolysis and the time from seeing a doctor to signing the informed consent for intravenous thrombolysis in the observation group were significantly shorter than those in the control group(P＜0.05).On the hospital admission,there was no significant difference in the National Institutes of Health Stroke Scale(NIHSS)score be-tween the two groups(P＞0.05);at the time points of 7 days after admission and 90 days after thrombolysis,the NIHSS scores of patients in the observation group were significantly lower than that in the control group(P＜0.05).There were no significant differences in the levels of serum glutathione peroxidase(GSH-Px)and malonaldehyde(MDA)between the two groups(P＞0.05);the level of serum superoxide dismutase(SOD)in the observation group was significantly higher than that in the control group(P＜0.05).One patient died in the control group,with a mortality rate of 3.33％;no patient died in the observation group.Conclusion Pre-hospital and intra-hospital collaborative therapeutic model can effectively improve the time efficiency of intravenous thrombolysis for patients with acute cerebral infarction,alleviate the neurological damage,and reduce degree of oxidative stress reaction and death risk.

Objective To observe the effect of pre-hospital and intra-hospital collaborative ther-apeutic model in the treatment of patients with acute cerebral infarction.Methods A total of 67 pa-tients with acute cerebral infarction were selected as the research objects,and they were divided into observation group(n=37)and control group(n=30)according to the therapeutic model.The con-trol group was conducted with the traditional therapeutic model,while the observation group was con-ducted with the pre-hospital and intra-hospital collaborative therapeutic model.The time efficiency of intravenous thrombolysis,early recovery of nerve function and oxidative stress indexes were compared between the two groups.Results There was no significant difference in the time from onset to visit between the two groups(P＞0.05);the time from seeing a doctor to thrombolysis and the time from seeing a doctor to signing the informed consent for intravenous thrombolysis in the observation group were significantly shorter than those in the control group(P＜0.05).On the hospital admission,there was no significant difference in the National Institutes of Health Stroke Scale(NIHSS)score be-tween the two groups(P＞0.05);at the time points of 7 days after admission and 90 days after thrombolysis,the NIHSS scores of patients in the observation group were significantly lower than that in the control group(P＜0.05).There were no significant differences in the levels of serum glutathione peroxidase(GSH-Px)and malonaldehyde(MDA)between the two groups(P＞0.05);the level of serum superoxide dismutase(SOD)in the observation group was significantly higher than that in the control group(P＜0.05).One patient died in the control group,with a mortality rate of 3.33％;no patient died in the observation group.Conclusion Pre-hospital and intra-hospital collaborative therapeutic model can effectively improve the time efficiency of intravenous thrombolysis for patients with acute cerebral infarction,alleviate the neurological damage,and reduce degree of oxidative stress reaction and death risk.

Objective : To analyze the pregnancy outcome of preimplantation genetic testing for aneuploidy (PGT-A) in cycles with different indications． Methods : The clinical information of 549 couples who underwent PGT-A were retrospectively analyzed．The cycles were divided into 6 groups according to the indication for PGT-A，namely : recurrent pregnancy loss group(n = 304) ，repeated implantation failure group(n = 57) ，advanced age group( ≥38 years old，n = 80) ，history of adverse pregnancy group(chorionic trisomy or adverse pregnancy，n = 24) ，male factor infertility group (n = 67) ，and abnormal sex chromosome number group (n = 17) ．The basic information，the number of retrieved oocytes，embryo biopsy result and pregnancy outcome were compared among different indication groups. Results: The average age and days of gonadotropin ( Gn) used among the six groups were statistically different (P＜0. 001) ．The average number of retrieved oocytes，the rate of good-quality embryos，mosaic embryos， abnormal embryos and normal embryos，the average ovarian sensitivity index ( OSI) among the six groups were statistically different (P = 0. 03，P ＜0. 001，P = 0. 03，P ＜0. 001，P ＜0. 001，P ＜0. 001 ) ．Advanced age group had the highest rate of abnormal embryos，the least average number of retrieved oocytes，the lowest OSI and the lowest rate of normal embryos．There were statistical differences in clinical pregnancy rate，ongoing pregnancy rate and cumulative pregnancy rate per oocyte retrieved cycle (P＜0. 001) among the six groups，but there were no statistical differences in clinical pregnancy rate，ongoing pregnancy rate per transfer cycle and cumulative pregnancy rate among the five groups except for the male factor infertility group． Conclusion PGT-A can detect euploid embryo to transfer thereby improving pregnancy efficiency．The advanced age women have normal embryo to transfer and can obtain a better pregnancy rate，which may shorten their time of“take- baby -home ”．At the same time， PGT-A can significantly improve the pregnancy outcome of those with male factor infertility.

Objective : To analyze the data related to the clinical outcome of preimplantation genetic testing (PGT) for double frozen , double biopsied blastocysts and double frozen , once biopsied blastocysts , in order to expand the existing data and provide some guidance for the clinical value and safety of PGT for frozen⁃thawed embryos . Methods : Retrospective analysis was made on the 38 PGT cycles of frozen⁃thawed blastocysts . According to the frequency of biopsy , cases in the study were divided into two groups : double frozen , double biopsy ( DFDB) group and double frozen , single biopsy ( DFSB) group . The freezing method was vitrification . Results : There were 24 patients in DFDB group , 34 blastocysts were not diagnosed in the last PGT cycle , 32 blastocysts survived after thawing , and the survival rate of thawed blastocysts was 94. 12% . After the second biopsy of these 32 blastocysts , genetic testing was performed , and all of them were definitely diagnosed , including 15 normal blastocysts (46. 88% ) and 17 abnormal blastocysts (53 . 13% ) . There were 14 patients in DFSB . The remaining 50 blastocysts in the last ICSI cycle were thawed and all blastocysts survived after thawing . Biopsy of these 50 blastocysts and genetic analysis showed that 47 blastocysts were diagnosed , including 9 normal blastocysts (18 . 00% ) , 28 abnormal blastocysts (56. 00% ) , 10 mosaic blastocysts (20. 00% ) , and 3 undiagnosed blastocysts (6. 00% ) . In DFDB group and DFSB group , 8 patients and 5 patients transferred the normal blastocystswhich all survivedafter thawing . There were 5 clinical pregnancies and 3 clinical pregnancies , respectively . One healthy live birth was obtained respectively in each group . Conclusion Acceptable pregnancy rate can be obtained whatever DFSB or DFDB blastocyst , which is of clinical value . However , due to the small sample size , we need to expand the sample size to further explore its safety .

Globally, the epidemic of diabetes mellitus has brought a series of health and economic burden, and the prevalence of diabetes mellitus in China is also rising. In recent years, with more insight into the mechanisms of diabetes mellitus, early diagnosis, accurate classification and effective treatment using genetic testing has been gained increasing attention. This article discusses the genetic susceptibility or pathogenicity genes of diabetes, and summarizes the progress of gene diagnosis in different types of diabetes.

Osteoarthritis (OA) is the most common form of arthritis and the leading cause of old age disability, affecting an estimated 302 million people worldwide. OA is seriously overlooked in the world. The awareness of OA and the popularization of standardized diagnosis and treatment are all lacking. Knees, hips, and hands are the most commonly affected joints in OA. Based on the experience of diagnosis and treatment, consensus and guidelines, we formulated this diagnosis and treatment standard in order to standardize the diagnosis and treatment of OA. We hope that our standard can reduce misdiagnosis and mistreatment and improve the prognosis of OA.

OBJECTIVES: To analyze the clinical characteristics of fecal severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid-positive in patients with coronavirus dasease 2019 (COVID-19) and to provide a scientific basis for the prevention and control of this disease. METHODS: The clinical data of 16 patients with fecal SARS-CoV-2 nucleic acid positive, who hospitalized in the North Branch of the First Hospital of Changsha (Changsha Public Health Rescue Center) from January to February 2020, were retrospectively analyzed. Their clinical manifestations, laboratory data and imaging data were summarized. RESULTS: Among the 16 patients, there were 9 males (56.25%) and 7 females (43.75%), the ratio of males to females was 1∶1.29. The age of onset was (43.3±14.6) years. There were 15 patients with contact history of Wuhan, 1 patient with contact history of local patient.Twelve patients were common type (75%), and 4 patients were severe type (25%). Clinical symptoms included fever in 14 patients (87.5%), cough in 12 patients (75%), shortness of breath in 5 patients (31.25%), pharyngalgia in 10 patients (62.5%), fatigue in 7 patients (43.75%), and diarrhea in 4 patients (25%). There were 14 patients (87.5%) with normal or decreased white blood cell count, 11 patients (68.75%) with decreased lymphocyte count, 15 patients (93.75%) with increased erythrocyte sedimentation rate, 13 patients (81.25%) with increased hypersensitivity C-reactive protein, 5 patients (31.25%) with increased procalcitonin, and 8 patients (50%) with increased serum ferritin in peripheral blood, and stool routine was basically normal. Compared with the common type, there was significant difference in the white blood cell and lymphocyte counts in the severe type (<0.01); the infection indicators, such as hypersensitivity C-reactive protein and serum ferritin, were significantly increased, with significant difference (all <0.01); but the procalcitonin and erythrocyte sedimentation rate was not significantly different (both >0.05). Chest CT mainly showed patchy shadows and interstitial changes. According to imaging examination, 4 patients (25%) showed unilateral pneumonia and 12 patients (75%) showed bilateral pneumonia. CONCLUSIONS The patients have the clinical symptoms of COVID-19, but gastrointestinal symptoms (such as diarrhea) are more common, and the changes of white blood cell count, lymphocyte count, hypersensitivity C-reactive protein, ferritin are more obvious in severe patients.The positivity of fecal nucleic acid suggests the possibility of digestive tract transmission of SARS-CoV-2, and fecal nucleic acid testing can be used as a routine testing method in clinical practice.
Adult ; Betacoronavirus ; isolation & purification ; C-Reactive Protein ; analysis ; China ; Coronavirus Infections ; diagnosis ; physiopathology ; Diarrhea ; virology ; Feces ; virology ; Female ; Ferritins ; analysis ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; physiopathology ; Retrospective Studies