In this study, the chloroplast genome of Camellia insularis Orel & Curry was sequenced using high-throughput sequencing technology. The results showed that the chloroplast genome of C. insularis was 156 882 bp in length with a typical tetrad structure, encoding 132 genes, including 88 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Codon preference analysis revealed that the highest number of codons coded for leucine, with a high A/U preference in the third codon position. Additionally, 67 simple sequence repeats (SSR) loci were identified, with a preference for A and T bases. The inverted repeat (IR) boundary regions of the chloroplast genome of C. insularis were relatively conserved, except for a few variable regions. Phylogenetic analysis indicated that C. insularis was most closely related to C. fascicularis. Yellow camellia is a valuable material for genetic engineering breeding. This study provides fundamental genetic information on chloroplast engineering and offers valuable resources for conducting in-depth research on the evolution, species identification, and genomic breeding of yellow Camellia.
Genome, Chloroplast/genetics* ; Phylogeny ; Plant Breeding ; Camellia/genetics* ; Chloroplasts/genetics*

Background Intraseasonal variation in acute health effects of extreme heat remains insufficiently investigated. Emergency ambulance calls (EACs) may offer timely insights into the population's health during such extreme heat events. Objective To analyze intraseasonal variation in the association between extreme heat and hourly EACs during summer in Dezhou City, Shandong Province, China. Methods We collected data on all-cause hourly EACs in Dezhou City from 2021 to 2022 and assigned hourly temperature and humidity data (with a spatial resolution of 0.0625° × 0.0625°) to call addresses. Summer in this study was defined as from June to September each year, with June to July considered as early summer and August to September as late summer. Extreme heat was defined as the 99th percentile of the temperature range during the summer. We employed a time-stratified case-crossover design using conditional logistic regression integrating distributed-lag nonlinear models to compare the association between extreme heat and the risk of hourly EACs in both early and late summer periods. Results A total of 80389 EACs were recorded in Dezhou City during the study period. The analysis revealed a U-shaped association between hourly ambient temperature and EACs during summer, with the most significant effect observed at lag 0-30 h. Using the optimal temperature of 20.0°C as a reference, the cumulative odds ratio (OR) (lag 0-120 h) for extreme heat was 1.55 (95%CI: 1.40, 1.71) throughout summer. The cumulative effect of extreme heat was higher in late summer (OR=2.38, 95%CI: 1.91, 2.97) than in early summer (OR=1.37, 95%CI: 1.22, 1.54) (P<0.0001). Additionally, individuals aged 60 years and above had a higher risk throughout summer (OR=1.98, 95%CI: 1.70, 2.30) compared to those under 60 years (OR=1.23, 95%CI: 1.06, 1.42) (P<0.0001). Conclusion Intraseasonal variation is observed in the strength of association between extreme heat and hourly EACs during summer in Dezhou City. The higher risk observed in late summer than in early summer indicates that repeated exposures to heat may escalate health risks, and older adults are more vulnerable.

SARS-CoV-2 is a new RNA virus affecting humans and spreads extensively throughout the world since its first outbreak in December,2019.Whether the transmissibility and pathogenicity of SARS-CoV-2 in humans after zoonotic transfer are actively evolving,and driven by adaptation to the new host and environments is still under debate.Understanding the evolutionary mechanism underlying epidemiological and pathological characteristics of COVID-19 is essential for predicting the epidemic trend,and providing guidance for disease control and treatments.Interrogating novel strategies for identifying natural selection using within-species polymorphisms and 3,674,076 SARS-CoV-2 genome sequences of 169 countries as of December 30,2021,we demonstrate with popula-tion genetic evidence that during the course of SARS-CoV-2 pandemic in humans,1)SARS-CoV-2 genomes are overall conserved under purifying selection,especially for the 14 genes related to viral RNA replication,transcription,and assembly;2)ongoing positive selection is actively driving the evolution of 6 genes(e.g.,S,ORF3a,and N)that play critical roles in molecular processes involving pathogen-host interactions,including viral invasion into and egress from host cells,and viral inhi-bition and evasion of host immune response,possibly leading to high transmissibility and mild symptom in SARS-CoV-2 evolution.According to an established haplotype phylogenetic relation-ship of 138 viral clusters,a spatial and temporal landscape of 556 critical mutations is constructed based on their divergence among viral haplotype clusters or repeatedly increase in frequency within at least 2 clusters,of which multiple mutations potentially conferring alterations in viral transmis-sibility,pathogenicity,and virulence of SARS-CoV-2 are highlighted,warranting attention.

Objective:To explore the epidemiologic characteristics and the possible risk factors of microtia in China. Meanwhile, the significant variables related to severe cases are integrated into a predictive nomogram.Methods:A total of 593 patients with congenital microtia from July 2015 to July 2018 were included. Patients conforming to congenital microtia with or without associated malformations were enrolled in this study, and patients with clear chromosomal syndromes were excluded. Questionnaire surveys were conducted among the parents to collect the demographic information and risk factors for exposure during perinatal period. Using Chi-Square and Fisher’s tests to statistically analyze the frequencies of variables. Univariate and multivariate logistic regression analysis were used to select variables related to severe cases for constructing nomogram. Concordance index (C-index), calibration plot, Hosmer-Lemeshow test, and receiver operating characteristics (ROC) curve were used to assess the nomogram model.Results:Of the patients, 456 (76.9%) were male and 137 (23.1%) were female. Right side was involved in 329 cases (55.5%), left side in 217 cases (36.6%) and both sides in 47 cases (7.9%). Among them, 16 cases were familial and the rest were sporadic. Multiple deformations were in 392 cases (66.1%). Maternal illness in early pregnancy( OR=2.205, 95% CI: 1.020-4.020)and parternal drinking history( OR=2.221, 95% CI: 1.329-3.677)were independent risk factors for severe microtia. While mother aged from 26 to 35 years old ( OR=0.507, 95% CI: 0.281-0.913; OR=0.258, 95% CI: 0.125-0.531) and father living in plain area( OR=0.512, 95% CI: 0.288-0.913)may be protective factors. All the significant predictors were combined into a predictive nomogram. The C-index was 0.703(95% CI: 0.646-0.760). The calibration plotshowed good performance of the nomogram, and the model passed Hosmer-Lemeshow goodness-of-fit test ( χ2=4.512, P=0.808). ROC curve analysis revealed a high sensitivity and specificity. Conclusions:The majority of microtia patients are male, sporadic, occur on right side, and often associated with other malformations. This nomogram predicting severe microtia based on multiple parental risk factors was with good discrimination and accuracy, which could provide scientific guidance for individualized prevention in clinical practice.

COVID-19 has swept globally and Pakistan is no exception.To investigate the initial introductions and transmissions of the SARS-CoV-2 in Pakistan,we performed the largest genomic epidemiology study of COVID-19 in Pakistan and generated 150 complete SARS-CoV-2 genome sequences from samples collected from March 16 to June 1,2020.We identified a total of 347 mutated positions,31 of which were over-represented in Pakistan.Meanwhile,we found over 1000 intra-host single-nucleotide variants(iSNVs).Several of them occurred concurrently,indicating possible interactions among them or coevolution.Some of the high-frequency iSNVs in Pakistan were not observed in the global population,suggesting strong purifying selections.The genomic epidemiology revealed five distinctive spreading clusters.The largest cluster consisted of 74 viruses which were derived from different geographic locations of Pakistan and formed a deep hierarchical structure,indicating an extensive and persistent nation-wide transmission of the virus that was probably attributed to a signature mutation(G8371T in ORF 1ab)of this cluster.Further-more,28 putative international introductions were identified,several of which are consistent with the epidemiological investigations.In all,this study has inferred the possible pathways of introduc-tions and transmissions of SARS-CoV-2 in Pakistan,which could aid ongoing and future viral surveillance and COVID-19 control.

Objective:To explore the epidemiologic characteristics and the possible risk factors of microtia in China. Meanwhile, the significant variables related to severe cases are integrated into a predictive nomogram.Methods:A total of 593 patients with congenital microtia from July 2015 to July 2018 were included. Patients conforming to congenital microtia with or without associated malformations were enrolled in this study, and patients with clear chromosomal syndromes were excluded. Questionnaire surveys were conducted among the parents to collect the demographic information and risk factors for exposure during perinatal period. Using Chi-Square and Fisher’s tests to statistically analyze the frequencies of variables. Univariate and multivariate logistic regression analysis were used to select variables related to severe cases for constructing nomogram. Concordance index (C-index), calibration plot, Hosmer-Lemeshow test, and receiver operating characteristics (ROC) curve were used to assess the nomogram model.Results:Of the patients, 456 (76.9%) were male and 137 (23.1%) were female. Right side was involved in 329 cases (55.5%), left side in 217 cases (36.6%) and both sides in 47 cases (7.9%). Among them, 16 cases were familial and the rest were sporadic. Multiple deformations were in 392 cases (66.1%). Maternal illness in early pregnancy( OR=2.205, 95% CI: 1.020-4.020)and parternal drinking history( OR=2.221, 95% CI: 1.329-3.677)were independent risk factors for severe microtia. While mother aged from 26 to 35 years old ( OR=0.507, 95% CI: 0.281-0.913; OR=0.258, 95% CI: 0.125-0.531) and father living in plain area( OR=0.512, 95% CI: 0.288-0.913)may be protective factors. All the significant predictors were combined into a predictive nomogram. The C-index was 0.703(95% CI: 0.646-0.760). The calibration plotshowed good performance of the nomogram, and the model passed Hosmer-Lemeshow goodness-of-fit test ( χ2=4.512, P=0.808). ROC curve analysis revealed a high sensitivity and specificity. Conclusions:The majority of microtia patients are male, sporadic, occur on right side, and often associated with other malformations. This nomogram predicting severe microtia based on multiple parental risk factors was with good discrimination and accuracy, which could provide scientific guidance for individualized prevention in clinical practice.

Objective:To find the chromosomal malfomations among microtia patients and the neighbouring genes of chromosomal aberrations or genes in the extra or deleted chromosome fragments would be screened to investigate the possible causative genes.Methods:According to the inclusion criteria, case group was selected from microtia patients referred to Plastic Surgery Hospital, Chinese Academy of Medical Science, between January 2012 and January 2014, and the control group was the normal people of similar age received plastic surgery in the same hospital in the same time who did not have any congenital genetic disease. Blood samples of two groups were collected, and genomic DNA was extracted, then copy number variation (CNV) analysis was performed in the two groups with gene chip technology and associated software for large fragment chromosomal malformations. The variations of chromosome copy number were recorded to further analyze the type and length of chromosome structure variation. The genes at the loci of break points were further screened referring to B allele frequency to interpret associated genes related to the occurrence of microtia. Fisher exact test were used for statistical analysis, and the difference was statistically significant ( P< 0.05). Results:942 patients with congenital microtia were included in the case group, 695 males and 247 females, aged (11.4±3.2) years; 1 802 normal controls, 1 290 males and 512 females, aged (11.6±4.9) years. Large chromosomal fragments variations were detected in 5 patients in chromosome in case group( P=0.003). The difference between the two groups was statistically significant( P=0.003). Three cases were found to carry an extra X chromosome. Among the 3 cases, one patient suffered from XXY karyotype and the other 2 patients X trisomy. Two cases were proved to be associated with chromosome structural variations. The malformations of the first case presented partial duplication of the long arm of chromosome 13 and 14. On searching for causative genes, OTX2, BMP4 and GSC were detected to be in the chromosome structural variations. The second case presented to be partial duplication of the long arm of chromosome 5. FGF pathway associated genes FGF18, FGFR4, FGF1 and BMP pathway associated genes FST, MSX2, SMAD5 were incorporated in the extra duplicated chromosome for possible gene dosage effect. Conclusions:The results indicated the possible association of chromosome abnormity and microtia and provide new insights in microtia-associated chromosome instability. Ten related genes were involved in the occurrence of microtia through various ways.

On January 22, 2020, China National Center for Bioinformation (CNCB) released the 2019 Novel Coronavirus Resource (2019nCoVR), an open-access information resource for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 2019nCoVR features a comprehensive integra-tion of sequence and clinical information for all publicly available SARS-CoV-2 isolates, which are manually curated with value-added annotations and quality evaluated by an automated in-house pipeline. Of particular note, 2019nCoVR offers systematic analyses to generate a dynamic landscape of SARS-CoV-2 genomic variations at a global scale. It provides all identified variants and their detailed statistics for each virus isolate, and congregates the quality score, functional annotation,and population frequency for each variant. Spatiotemporal change for each variant can be visualized and historical viral haplotype network maps for the course of the outbreak are also generated based on all complete and high-quality genomes available. Moreover, 2019nCoVR provides a full collection of SARS-CoV-2 relevant literature on the coronavirus disease 2019 (COVID-19), including published papers from PubMed as well as preprints from services such as bioRxiv and medRxiv through Europe PMC. Furthermore, by linking with relevant databases in CNCB, 2019nCoVR offers data submission services for raw sequence reads and assembled genomes, and data sharing with NCBI. Collectively, SARS-CoV-2 is updated daily to collect the latest information on genome sequences, variants, hap-lotypes, and literature for a timely reflection, making 2019nCoVR a valuable resource for the global research community. 2019nCoVR is accessible at https://bigd.big.ac.cn/ncov/.

Objective:To find the chromosomal malfomations among microtia patients and the neighbouring genes of chromosomal aberrations or genes in the extra or deleted chromosome fragments would be screened to investigate the possible causative genes.Methods:According to the inclusion criteria, case group was selected from microtia patients referred to Plastic Surgery Hospital, Chinese Academy of Medical Science, between January 2012 and January 2014, and the control group was the normal people of similar age received plastic surgery in the same hospital in the same time who did not have any congenital genetic disease. Blood samples of two groups were collected, and genomic DNA was extracted, then copy number variation (CNV) analysis was performed in the two groups with gene chip technology and associated software for large fragment chromosomal malformations. The variations of chromosome copy number were recorded to further analyze the type and length of chromosome structure variation. The genes at the loci of break points were further screened referring to B allele frequency to interpret associated genes related to the occurrence of microtia. Fisher exact test were used for statistical analysis, and the difference was statistically significant ( P< 0.05). Results:942 patients with congenital microtia were included in the case group, 695 males and 247 females, aged (11.4±3.2) years; 1 802 normal controls, 1 290 males and 512 females, aged (11.6±4.9) years. Large chromosomal fragments variations were detected in 5 patients in chromosome in case group( P=0.003). The difference between the two groups was statistically significant( P=0.003). Three cases were found to carry an extra X chromosome. Among the 3 cases, one patient suffered from XXY karyotype and the other 2 patients X trisomy. Two cases were proved to be associated with chromosome structural variations. The malformations of the first case presented partial duplication of the long arm of chromosome 13 and 14. On searching for causative genes, OTX2, BMP4 and GSC were detected to be in the chromosome structural variations. The second case presented to be partial duplication of the long arm of chromosome 5. FGF pathway associated genes FGF18, FGFR4, FGF1 and BMP pathway associated genes FST, MSX2, SMAD5 were incorporated in the extra duplicated chromosome for possible gene dosage effect. Conclusions:The results indicated the possible association of chromosome abnormity and microtia and provide new insights in microtia-associated chromosome instability. Ten related genes were involved in the occurrence of microtia through various ways.

Objective To explore the monitoring of cerebrospinal fluid (CSF) dynamics in a model of brain herniation induced by acute intracranial hypertension in Guangxi Bama-Mini pigs by phasecontrast cine magnetic resonance imaging (PC cine MRI).Methods Femoral artery blood were extracted from 10 pigs,and injected into the frontal and temporal parietal lobe to make a model of brain herniation induced by acute intracranial hypertension.The mean arterial blood pressure (MAP),intracranial pressure (ICP),and cerebral perfusion pressure (CPP) were monitored.Routine T1WI,T2WI,coronal,sagittal and cerebrospinal fluid flow sequence (fast PC cine slice) which positioned on the cervical 3 (C3) vertebral body as the center and perpendicular to the spinal scans were performed on all experimental animals before and after blood injection with 3.0T Magnetic Resonance Imaging.The ICP,MAP,CPP,the absolute values of CSF peak flow velocity and the absolute value of carotid peak flow velocity before and after blood injection were compared.Results The ICP,MAP,CPP,and the absolute value of CSF peak flow velocity before injection of autologous arterial blood were statistically significant as compared with those after blood injection [(6.80±2.044) mmHg vs (52.20±1.619) mmHg,(76.80±7.068) mmHg vs (142.80±12.399) mmHg,(70.00±6.074) mmHg vs (90.50±12.250) mmHg,and the absolute value of CSF peak flow velocity was (243.20±77.671) mm/s vs (201.40±55.482) mm/s,respectively,P＜0.01].The absolute value of the peak velocity of the carotid artery before blood injection was not statistically significant compared with that after blood injection [(876.80±239.908) mm/s vs (799.40±241.829) mm/s,P＞0.05].Conclusion After the formation of brain herniation induced by acute intracranial hypertension,the CSF flow in the C3 level spinal canal showed a low dynamic change,and the CSF flow velocity waveform was disordered and malformed.The non-invasive measurement of CSF dynamics by PC cine MRI can provide an important basis for the change of CSF dynamics in the model of brain herniation induced by acute intracranial hypertension,and provide a theoretical basis for further research on damage control neurosurgery in the future.