Objective:To assess the value of optical genome mapping (OGM) for the detection of chromosomal structural abnormalities including ring chromosomes, balanced translocations, and insertional translocations.Methods:Clinical data of four patients who underwent pre-implantation genetic testing concurrently with OGM and chromosomal microarray analysis at the Center of Reproductive Medicine of the Sixth Affiliated Hospital of Sun Yat-sen University from January to October 2022 due to chromosomal structural abnormalities were selected as the study subjects. Some of the results were verified by multi-color fluorescence in situ hybridization. Results:The OGM has successfully detected a balanced translocation and fine mapped the breakpoints in a patient. Among two patients with insertional translocations, OGM has provided more refined breakpoint locations than karyotyping analysis in a patient who had chromosome 3 inserted into chromosome 6 and determined the direction of the inserted fragment. However, OGM has failed to detect the chromosomal abnormalit in a patient with chromosome 8 inserted into the Y chromosome. It has also failed to detect circular signals in a patient with ring chromosome mosaicism.Conclusion:OGM has successfully detected chromosomal structural variations in the four patients and provided assistance for their diagnosis.

Objective:To compare the results of invasive prenatal diagnosis and preimplantation genetic testing (PGT) and explore the underlying mechanism.Methods:Clinical data of pregnant women undergoing PGT and invasive prenatal diagnosis at the Sixth Affiliated Hospital of Sun Yat-sen University from January 2019 to December 2022 were collected. The results of PGT and invasive prenatal diagnosis were compared, and the outcomes of pregnancies were followed up. This study has been approved by the Medical Ethics Committee of the the Sixth Affiliated Hospital of Sun Yat-sen University (No. 2022SLYEC-491).Results:A total of 172 couples were included in this study, and 26 non-targeted variants were discovered upon prenatal diagnosis, including 10 cases (38.5%) by chromosomal karyotyping, 15 (57.7%) by chromosomal microarray analysis (CMA), and 1 (3.8%) by whole exome sequencing. The 10 karyotypic anomalies had included 6 chromosomal polymorphisms, 2 chromosomal mosaicisms, 1 paternally derived translocation, and 1 missed maternal chromosomal inversion. CMA has identified 15 copy number variations (CNVs), which included 11 microdeletions and microduplications, 3 loss of heterozygosity, and 1 low-level mosaicism of paternal uniparental disomy. One CNV was classified as pathogenic, and another one was likely pathogenic, whilst the remaining 13 were classified as variants of uncertain significance. Therefore, 8.7% of CNVs was detected by invasive prenatal diagnosis after PGT. 92.3% (24/26) of the non-targeted variants have been due to technological limitations of next-generation sequencing (NGS).Conclusion:Invasive prenatal diagnosis after PGT can detect non-targeted variants, which may further reduce the incidence of birth defects.

Objective:To investigate the molecular epidemiological characteristics of coxsackievirus A16 (CVA16) in Fujian province from 2020 to 2023.Methods:The epidemiological characteristics of CVA16 associated hand, food and mouth disease (HFMD) in Fujian province from 2020 to 2023 was analyzed. The complete VP1 gene of CVA16 was amplified by RT-PCR and then sequenced, and genetic evolution was analyzed by MEGA X and other softwares.Results:From 2020 to 2023, there were 13 120 cases of HFMD in Fujian province, and the proportion of HFMD which caused by CVA16 was 16.5% (2 160/13 120). From 2020 to 2023, the proportion of accounted cases was 4.7% (94/2 019), 14.1% (457/3 243), 47.6% (1 521/3 199) and 1.9% (88/4 659) respectively. HFMD caused by CVA16 was mainly concentrated in children aged 1 to 5 years, and most of them were 3 years old. The genetic evolution and genotype analysis of 92 complete VP1 gene sequences obtained from 2020 to 2023 showed that the genetic distance between CVA16 strains in Fujian province and the prototype strain was far away. The CVA16 genotype in Fujian province from 2020 to 2023 has three clusters of B1a, B1b and B1c, among which the composition ratio of B1a and B1b in Fujian province in 2020 was 40% and 60% respectively. In 2021, B1a and B1b accounted for 81.8% and 18.2% respectively. Only B1a in 2022; in 2023, there were B1a, B1b and B1c, which respectively accounted for 44.4%, 7.4% and 48.2%. During the period from January to September, B1a was the main cluster. After October we observed an emergence of B1c cluster, which had never been found in Fujian province and was rare in China, was detected and became the dominant cluster.Conclusions:The evolutionary cluster of CVA16 dominant changed from B1b in 2020 to B1a in 2021-2023. After October 2023, the newly discovered B1c became the dominant cluster in Fujian province.

Objective:To investigate the protective effects of Huoxue Rongluo Decoction on hypoxic-ischemic brain damage (HIBD) rats; To clarify the expression of AMPK signaling pathway during HIBD injury and repair.Methods:On the basis of the hypoxia ischemia reperfusion model, an improved hypoxia bottle was used to establish a full-term HIBD rat model. Totally 60 successfully modeled mice were divided into model group and TCM group using a random number table method, with 30 mice in each group and 30 mice in a control group. Two hours before surgery, TCM group was orally administered with Huoxue Rongluo Decoction 1.17 g/100 g (equivalent dose). The control group and model group were orally administered with equal volume of physiological saline. Two hours after surgery, it was orally administered again, twice a day, for consecutive 5 d. Behavioral tests (convulsion assessment, Longa score, suspension test, water maze test) were used to evaluate the motor nerve function and long-term learning ability of young rats in different groups. HE staining and electron microscopy were used to observe the pathological changes of brain tissue and mitochondrial damage. Evans blue (EB) staining and brain water content measurement were used to detect blood-brain barrier permeability and brain edema. The expression of AMPK signaling pathway related genes in brain tissues of different groups of young rats was detected by PCR array technique.Results:Compared with the model group, the convulsions of the rats in the TCM group were improved, the Longa score was significantly reduced, the grip strength test score and suspension time were significantly improved, the escape latency was significantly shortened, and the number of crossing platforms significantly increased ( P<0.05); the degree of neuronal and mitochondrial damage in the TCM group was reduced, and cerebral vascular permeability and brain water content were significantly reduced ( P<0.01). The results of PCR array showed that compared with the control group, 2 genes were significantly up-regulated and 12 genes were significantly down-regulated in the brain tissue of the model group; compared with TCM group, 15 genes were relatively down-regulated in the model group. Conclusion:Huoxue Rongluo Decoction can significantly reduce the degree of brain injury after HIBD, improve the motor nerve function and long-term learning and cognitive ability of model rats, and its damage repair mechanism is related to the expression regulation of AMPK signaling pathway related genes.

Objective:To analyze the clinically acceptable and reproducible bladder and rectum volumes of prostate cancer patients during radiotherapy under bladder and bowel preparation, aiming to provide quantitative indicators for bowel and bladder preparation before and after radiotherapy.Methods:Clinical data of 275 prostate cancer patients with strict bladder and bowel preparation and completion of whole course radical radiotherapy at Sun Yat-sen University Cancer Center from April 2015 to December 2020 were retrospectively analyzed. Patients were scanned with cone beam CT (CBCT) before each treatment and the setup error was recorded. Sixty-six patients were selected by simple random sampling and the bladder and rectum on daily CBCT was outlined using MIM software. The relationship between the ratio of daily bladder or rectum volume to the planned bladder or rectum volume (relative value of volume) and setup error was analyzed. Quantitative data were expressed as mean±SD. Normally distributed data were analyzed by paired t-test while non-normally distributed data were assessed by Kruskal-Wallis test.Results:The bladder and rectum volume on planning CT were (370.87±110.04) ml and (59.94±25.07) ml of 275 patients. The bladder and rectum volumes on planning CT were (357.51±107.38) ml and (65.28±35.37) ml respectively of the 66 selected patients with 1611 sets of CBCT images. And the bladder and rectum volumes on daily CBCT were (258.96±120.23) ml and (59.95 ± 30.40) ml. The bladder volume of patients was decreased by 3.59 ml per day on average during the treatment and 0.37 ml for the rectum volume. According to the bladder volume on planning CT, all patients were divided into three groups: <250 ml, 250-450 ml and >450 ml groups. The relative value of volume in the 250-450 ml group during the course of radiotherapy was the smallest. And the setup error in the superior and inferior (SI) direction was (0.28±0.24) cm and (0.19±0.17) cm in the left and right (LR) direction, significantly lower than those in the other two groups (both P≤0.027). According to the rectum volume on planning CT, all patients were divided into four groups: <50 ml, 50-<80 ml, 80-120 ml and >120 ml groups. The <50 ml group had the smallest relative value of volume during radiotherapy, and the setup error in the SI direction was (0.26±0.22) cm and (0.24±0.22) cm in the anterior and posterior (AP) direction, significantly smaller than those in the other groups (both P≤0.003). The setup errors in the SI, LR, AP directions of the enrolled 66 patients were (0.30±0.25) cm, (0.20±0.18) cm and (0.28±0.27) cm, respectively. Among them, the relative value of bladder volume in the AP direction was (0.73±0.37) in the setup error <0.3 cm group, which was statistically different from those in the setup error 0.3-0.5 cm and >0.5 cm groups (both P<0.05). Conclusion:Under the bladder and bowel preparation before planning CT, the appropriate bladder and rectum volumes are in the range of 250-450 ml and <50 ml, which yields higher reproducibility and smaller setup error.

Objective:To investigate the outcomes and influencing factors of newly diagnosed prediabetic subjects aged 40 years and above in Guiyang.Methods:A total of 10 015 residents aged 40 years and above were recruited from the Yunyan community, Guiyang, from May to August 2011. Physical examination, laboratory measurements, and questionnaires were conducted. The follow-up survey was conducted in July 2014. A total of 2 530 newly diagnosed prediabetic subjects at baseline were included in the analysis.Results:The 3-year cumulative morbidity of diabetes mellitus was 14.3%, and the risk of diabetes mellitus in combined impaired fasting glucose(IFG)and impaired glucose tolerance(IGT)groups was significantly higher than that in isolated IFG(i-IFG)or isolated IGT(i-IGT)group( P<0.01). High baseline fasting plasma glucose, 2 h plasma glucose, and HbA 1C levels were the independent risk factors for the development of diabetes( OR=1.836, 95% CI 1.374-2.454; OR=1.398, 95% CI 1.261-1.550; OR=2.526, 95% CI 1.804-3.538, all P<0.01)and the inhibitory factors for reversion to normal glucose tolerance( OR=0.511, 95% CI 0.409-0.638; OR=0.715, 95% CI 0.661-0.774; OR=0.638, 95% CI 0.500-0.816, all P<0.01). High level of high density lipoprotein-cholesterol(HDL-C)was an promoting factor for reversion to normal glucose tolerance( OR=1.306, 95% CI 1.017-1.678, P=0.036). Subjects in the highest tertile of baseline HbA 1C level and body mass index(BMI)change before and after follow-up(ΔBMI=follow-up BMI minus baseline BMI)had a higher risk of diabetes mellitus than those in the lowest tertile( OR=2.398, 95% CI 1.733-3.322; OR=2.402, 95% CI 1.859-3.105, both P<0.01). The risk of diabetes mellitus in the significant weight loss group was reduced by 40.4% compared with the non-significant weight loss group when the subjects were divided into two groups according to the cutoff of the lower tertile of ΔBMI( RR=0.596, 95% CI 0.463-0.766, P<0.01). Conclusion:The risk of diabetes mellitus in combined IFG/IGT group was significantly higher than that in i-IFG or i-IGT group. High baseline fasting plasma glucose, 2 h plasma glucose, and HbA 1C levels were the independent risk factors for the development of diabetes. High level of HDL-C was an promoting factor for reversion to normal glucose tolerance. Weight loss can significantly reduce the risk of progression to diabetes in individuals with prediabetes.

Objective To establish and preliminarily apply an effective PCR assay for detection of Tupaia(tree shrew)paramyxovirus(TPMV). Methods Using TPMV genomic DNA from NCBI GenBank, bases 8231 to 8720 were synthesized and inserted into a plasmid as a positive standard. One primer pair was designed based on this sequence. In total,60 respiratory swabs and 12 lung tissues from the tree shrews were tested in this PCR assay. Results A PCR method for detection of TPMV was successfully established,with high specificity and sensitivity of 11.5 × 10 -5μg/mL. PCR result testing 60 respiratory swabs and 12 lung tissues were negative. Conclusions PCR for detecting TPMV has good specificity and high sensitivity and can be used for conventional tree shrew paramyxovirus detection.

Objective Four different methods were examined to identify a safer and more reliable method for tail vein punctures in C57BL/6 mice. Methods In total,320 mice were randomly divided into four groups: a blank group, incandescent lamp baking method group,three-line method group,and combined method group. Blood samples were taken from the left or right peripheral vein of puncture mice. Puncture success rate of each group was recorded. SPSS 13.0 software was used to compare statistical difference among groups. Results Compared with the blank group,success rates of the other three methods were significantly higher(P < 0.001). Further, the three-line method was better than the incandescent lamp baking method(P< 0.001). The success rate of the combined method was significantly higher than the three-line and incandescent lamp baking methods(P< 0.001). Conclusions The combined method greatly improved the success rate of tail vein punctures in C57BL/6 mice. This method is more reliable and should be more widely used in the future.

Objective To understand the characteristics of minipigs infected withJapanese encephalitis virus(JEV).Methods After the brain tissues were treated, the pig brain tissue treatment solution was inoculated with BHK21 cells.Then, virus culture,indirect immunofluorescence assay, neutralization test, electron microscopic observation, and reverse transcription-polymerase chain reaction (RT-PCR) amplification of the new isolate E segment and PrM segment nucleotide sequence were performed and the genotype was identified.Results BHK21 cells were inoculated into 25 pigbrain tissues.Among them, three tissue-treated fluid couldinduce shrinkage and aggregation of BHK21 cells, and immunofluorescence staining showed strong green fluorescence response.The results of neutralization test showed that the neutralization titer of these three new isolates was 1:64, and the size of the virus particles was about 40nm under the electron microscope.The homology of both RT-PCR product sequencing results and E-segment of vaccine strain were 95%.Three new isolates were type GIII JEV.Conclusion The results ofthisstudydemonstrate that there is G III type Japanese encephalitis virus infection in the minipig farm.

Objective To verify the detection ability of experimental animal quality detection laboratories in China for Staphylococcus aureus.Methods The testing samples for Staphylococcus aureus detection were prepared by bacterial culture, homogeneity test and stability test, according to the study plan approved by CNAS.Then the samples and operation instruction were sent to the participant laboratories.The detection reports from these laboratories should be submitted before the deadline expires, and the collected data were summarized and analyzed.Results There were 28 laboratories which joined to this test plan.Among them 22 laboratories ( 78.57%) achieved satisfactory test results, and six laboratories (21.43%) had unsatisfactory test results.27 Laboratories used the national standard detection assay, while only one labo-ratory used PCR assay.Conclusions Most of experimental animal quality testing laboratories in China have sufficient pro-ficiency in detection of Staphylococcus aureus.The obtained information are very helpful for the laboratory ability verification testing in future.