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.

The gastrointestinal tract is an extremely complex ecosystem,and a healthy and intact intestinal mucosal barrier is the basic defense against the translocation of harmful substances.The impaired function of the intestinal barrier in acute-on-chronic liver failure(ACLF)is further aggravated by the dysregulation of intestinal microecology,which leads to bacterial translocation and endotoxemia.Maintaining the normal function of the intestinal mucosal barrier is important for the treatment of ACLF.This paper reviews the function of the normal intestinal mucosal barrier,the relationship between the damaged intestinal barrier and ACLF,and the impact of the damaged intestinal barrier on ACLF progression.

Objective To investigate the clinical effects and pregnancy outcomes of using luteal phase long protocol and GnRH antagonist protocol in patients with polycystic ovary syndrome(PCOS)who have failed their first GnRH antagonist protocol therapy.Methods The clinical data of 163 PCOS patients who underwent IVF/ICSI-ET were retrieved.After the failure of their first GnRH antagonist protocol treatment,they were divided into two groups in the second controlled ovarian hyperstimulation(COH)cycle:Luteal phase long protocol group(n=95)and Gn-RH antagonist protocol group(n=68).A retrospective analysis and comparison of basic clinical data,clinical and laboratory indicators,and pregnancy outcomes between two groups were conducted.Results ① There was no sta-tistically significant difference in basic clinical indicators between two group except LH.② Compared the first and second cycle treatments of patients in the luteal phase long protocol group,the initiation dose of gonadotropin(Gn),total number of Gn days,total Gn usage,estradiol(E2)on the day of hCG injection,number of retrieved eggs,oocyte maturation rate,2PN fertilization rate,2PN cleavage rate,blastocyst formation rate,high-quality blas-tocyst formation rate,and moderate to severe OHSS rate were significantly higher than those in the first GnRH an-tagonist cycle(P＜0.05).The GnRH antagonist protocol group also showed similar improvements.③ The com-parison of the second COH cycle between two groups showed that the total number of Gn days,total Gn usage,and total Gn cost in the luteal phase long protocol group were significantly higher(P＜0.05),while the E2 and LH on the day of hCG injection,and the maturation rate of eggs were significantly lower than those in the GnRH antagonist protocol group(P＜0.05).However,there was no statistically significant difference in the number of retrieved eggs,2PN fertilization,2PN cleavage,blastocyst formation rate,high-quality blastocyst formation rate,and OHSS rate between the two groups;④ The comparison of fresh transplantation cycles for the second COH cycle between the two groups showed that the luteal phase long protocol fresh transplantation rate,implantation rate,clinical preg-nancy rate,and live birth rate were slightly higher than those of the GnRH antagonist protocol group,but the differ-ence was not statistically significant.Comparing the outcomes of pregnancy following the initial frozen-thawed em-bryo transfer(FET)between two groups,the biochemical pregnancy rate and clinical pregnancy rate of the GnRH antagonist protocol group were higher than those of the luteal phase long protocol group(P＜0.05).However,no significant statistical variations were found in implantation rate,live birth rate,neonatal gestational age,and birth weight.Conclusion For PCOS patients who fail the first GnRH antagonist protocol,an appropriate increase in the initiating dose and usage of Gn can achieve satisfactory pregnancy outcomes with both protocols.Compared with change to a luteal phase long protocol,reusing the GnRH antagonist protocol still maintains its long-standing advan-tages,such as shorter total Gn days,lower costs,and better patient compliance.

Objective To explore the technology frontiers for neuroblastoma treatment from the perspective of patent citation network. Methods Through patent analysis for neuroblastoma treatment, highly cited patents and highly cited papers in the citation network were taken as the research objects. The title and abstract of the citing patents were analyzed by text clustering to identify the technology frontiers. Through social network analysis, the core patents were identified from the indices of degree centrality, betweenness centrality, closeness centrality, and eigenvector centrality. Results A total of 6240 patent applications for neuroblastoma treatment were found, including 71304 patent citations and 88698 journal-article citations. Four technology frontiers were identified based on patent citation network, namely, drug target, drug design, tumor-indication expansion, and gene-expression regulation. Three technology frontiers were identified based on journal-article citation network. They were drug target, drug design, and tumor-indication expansion. Conclusion The development of technology for neuroblastoma treatment continues to be active. Drug target and drug design are the most important technology frontiers. This study could provide certain reference for neuroblastoma treatment from the perspective of information science.

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 .

Objective : To develop a prediction model for live birth based on the basic characters of patients after transcervical resection of adhesions (TCRA) undergoing frozen⁃thawed embryo transfer (F⁃ET) . Methods : A total of 491 cycles who went F ⁃ET after TCRA were included. The cycles were randomly divided into training (n = 347) and validation (n = 144) cohorts at 7 ∶ 3 ratio. Relevant factors of live birth were selected via logistic regression analysis based on the data from patients in the training cohort. Multivariate logistic regression analysis was used to establish a nomogram , and then this predict model was calibrated and verified. Results : Age , body mass index (BMI) , miscarriages and endometrial thickness were significantly related to the live birth. The statistical nomogram was built with the area under the ROC cu2rve for the training cohort was 0. 713 (95% CI: 0. 519 - 0. 815) , Hosmer⁃Lemeshow goodness of fit test were χ = 7. 062 , P = 0. 530 , and the slope of calibration curve was close to 1. AUC of validation cohort was 0. 609 (95% CI: 0. 660 - 0. 561) , presenting a well⁃pleasing goodness⁃of⁃fit and stability in this model. Conclusion Age , BMI , miscarriages and endometrial thickness can better predict the probability of live birth undergoing F ⁃ET after TCRA.

Objective:By building a COVID-19 related scientific research progress monitoring system, to mitigate the information needs of different stakeholders such as government decision-makers, enterprises and scientific research institutions, provide reference for global scientific research progress monitoring for possible public health emergencies in the future.Methods:Literature review and expert consultation were adopted to systematically sort out the data sources of journal papers, preprints and other scientific papers, formed their respective monitoring catalogs and determined the tracking path.Results:COVID-19-related papers mainly include journal papers and preprints. The data source monitoring range of journal papers includes the official websites of top journals in biomedical and natural sciences, the database Web of Science and Science Direct, and the monitoring scope of the preprint platform includes medRxiv, bioRxiv, and SSRN, etc. In addition, the virological websites virological and nextstrain should also be monitored.Conclusions:COVID-19-related scientific research progress monitoring system can be used for daily monitoring to provide support for decision-making and deployment of prevention and control work.

OBJECTIVE: In this study,computational modelling was used to study whether different conduction pathways have impact on the AF initiation and maintenance based on a human atrial model. METHODS: MRI images from one patient who has atrial fibrillation were used to reconstruct the human atrial anatomical model.The number of MRI images was 112.In order to study the effect of different conduction pathways to AF,4 different atrial models were generated in this study,which were biatrial conduction via only LFO;biatrial conduction via only CS;biatrial conduction via both LFO and CS;model without biatrial conduction. RESULTS: In the case of biatrial conduction simulation via only LFO,the same re-entries were induced as in the LFO and CS case.But for the two cases of only CS conduction and both LFO and CS cannot conduct electricity,one stimulus site induced unsustained re-entry which only lasted for 2 cycles,and then terminated.The other two sites induced the same re-entries as in the previously described two cases,which were biatrial conduction simulation via both LFO and CS and only LFO. CONCLUSIONS Our simulation showed that these conduction pathways played a minor role in the re-entry initiation and maintenance,the possible explanation is that all the re-entries induced in our simulation is local drivers and maintained by fibrosis.
Atrial Fibrillation ; Catheter Ablation ; instrumentation ; Heart Atria ; Heart Rate ; Humans

INTRODUCTION: Paraquat (PQ) intoxication is frequently associated with a high mortality rate. No specific treatment has been shown to reduce mortality in victims within the first 72 hours. We investigated the protective effects of rapamycin (Rapa) against PQ-induced toxicity in a zebrafish model. METHODS: To determine the maximum nonlethal concentration (MNLC) and lethal concentration 50 (LC50) of Rapa, zebrafish were treated at 2-5 days post fertilisation (dpf) and their mortality was recorded every 24 hours. At 5 dpf, the zebrafish were treated with PQ 100 µg/mL or PQ+Rapa (MNLC, 1/3 MNLC or 1/9 MNLC) for 72 hours, and the rate of survival was recorded every 24 hours. Reverse transcription-polymerase chain reaction was used to test the signalling pathway of mTOR (mammalian target of rapamycin). RESULTS: MNLC and LC50 of Rapa were determined to be 6.7 µg/mL and 28.9 µg/mL, respectively. At 48 hours, the PQ+Rapa groups had much lower mortality than the PQ group. The rates of survival of the PQ+Rapa groups were 43.33% (MNLC), 53.89% (1/3 MNLC) and 44.45% (1/9 MLNC), as compared to 19.45% in the PQ group, with the 1/3 MNLC group showing the highest rate of survival (p < 0.001). atg1 was slightly activated in the PQ group. In the PQ+Rapa groups, the expression of atg1 was markedly increased, suggesting strengthening of the autophagy process. CONCLUSION Rapa can increase the rate of survival of PQ-intoxicated zebrafish by inhibiting mTOR complex 1 and activating autophagy. Rapa could be an alternative first-line drug in the treatment of PQ poisoning.

Objective To use the liquid protein combined with MALDI-TOF-MS for screening the serum differential peptides markers in lung adenocarcinoma patients and to establish the lung adenocarcinoma diag-nosed prediction model for founding the potential markers for the diagnosis of lung adenocarcinoma.Methods 37 patients with lung adenocarcinoma and 33 healthy subjects and benign lung disease which were made up in control group were collected,in the two groups the age and the sex were matched.The two groups were ran-domly divided into training group(30 cases of lung adenocarcinoma,26 cases of control)and test group(7 ca-ses of lung adenocarcinoma,7 cases of control)according to 3:1.T he differential diagnosis of lung adenocarci-noma and control group was performed by liquid chip-time-of-flight mass spectrometry and software ClinPro-Tools 3.0 to establish a prediction model of lung adenocarcinoma.The diagnostic model was validated by using serum samples from the test group to assess the diagnostic efficacy of the model.Results Nine peptide peaks with significant differences(P<0.05)were obtained by ClinProTools 3.0 software analysis.The up-regulated peaks in lung adenocarcinoma(m/z)were 8 976.5,4 469.05,4 966.78,8 925.5,4 531.05,and the down-reg-ulated m/z were 3 304.44,8 594.76,3 266.82,3 195.52.According to the genetic algorithm(GA),the lung ad-enocarcinoma diagnosis and prediction model was established.The overall recognition ability of the model was 94.49%.The model was evaluated by the test group.The results showed that the sensitivity of the model was 100.0% and the specificity was 85.7%.Conclusion Among lung adenocarcinoma patients,serum benign lung disease and healthy,there are differences in the serum peptide.T he use of differential peptide peaks to estab-lish lung adenocarcinoma diagnostic prediction model for the early diagnosis of lung adenocarcinoma provides a new method.