Objective To use direct dilution method to pretreat human urine, and to determine 19 elements in human urine using inductively coupled plasma mass spectrometry, and to evaluate the uncertainty of the entire experimental process. Methods The relevant mathematical models were established according to JJF 10591-2012 ^“Evaluation and Expression of Measurement Uncertainty^” and CNAS-GL006 ^“Guidance for Evaluation of Uncertainty in Chemical Analysis^”. Taking molybdenum as an example, the uncertainty sources in the determination of 19 elements in human urine by inductively coupled plasma method were analyzed and evaluated, including sample pretreatment, sample repeated measurement, standard solution preparation and standard curve fitting. Results The extended uncertainty of molybdenum in human urine is 2.12μg/L, and the measurement result of molybdenum is (44.8±2.12)μg/L. The measurement result of 19 elements in human urine ranges from less than the detection limit to 601μg/L, and the extended uncertainty range is 0.38~33.6μg/L.Conclusion It was found from the calculation that the uncertainty of the determination result was mainly affected by the uncertainty of the sample repeated measurement and the standard curve fitting. By adjusting the range of standard curve and increasing the number of parallel sample measurement, the uncertainty was reduced and the quality of detection was improved.

ObjectiveThis study observes the intervention effect of Longmu Piyan prescription on oxidative stress in BALB/c mice with atopic dermatitis （AD） induced by 2，4-dinitrochlorobenzene （DNCB） and explores its mechanism. MethodThe AD model was established using the method of DNCB sensitization on the back skin of BALB/c mice. Forty male BALB/c mice were randomly divided into a blank group， a model group， a vitamin C control group （0.5×10^-3 mg·kg^-1）， and a Longmu Piyan prescription group （26 g·kg^-1）. Except for the blank group， other groups were sensitized with different concentrations of DNCB on the back to induce AD， and the blank group was treated with matrix coating. The gastric administration was started on the seventh day after sensitization with 2% DNCB and on the 24th day after sensitization with 0.2% DNCB continuously for 21 days. The changes in skin lesions of each group were directly observed after the experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin （IL）-4， tumor necrosis factor （TNF）-α， immunoglobulin E （IgE）， and reactive oxygen species （ROS） in the serum of each group. The total antioxidant capacity determination kit-trace method (ABTS method) was used to measure the level of total antioxidant capacity （TAOC） in serum. The Hematoxylin eosin （HE） staining method was used to observe the pathological and morphological changes of the skin lesion site. The immunohistochemical method was used to detect the expression of thymic stromal lymphopoietin （TSLP） in the skin lesion site. Western blot was used to detect the expression of filaggrin （FLG） in the dorsal skin lesions. ResultThe results showed that compared with the blank group， the skin lesion score of the model group mice was significantly increased （P<0.01）， and HE staining showed characteristic pathological changes of AD in the skin lesion site. At the same time， the expression of TSLP in the skin lesion was significantly increased， and that of FLG was reduced （P<0.05）. The levels of TNF-α， IL-4， IgE， and ROS in serum increased， while the activity of TAOC decreased （P<0.01）. Compared with the model group， the Longmu Piyan prescription group showed a significant decrease in skin lesion scores and a significant improvement in skin lesion pathology. At the same time， the expression of TSLP decreased， and the expression of FLG increased in the skin lesions （P<0.05）. In addition， compared with the model group， the serum levels of TNF-α， IL-4， IgE， and ROS also decreased to varying degrees （P<0.05，P<0.01）， and TAOC activity increased in the Longmu Piyan prescription group （P<0.01）. ConclusionThere is a significant correlation among the degree of oxidative stress， the severity of skin lesions in AD， and the levels of inflammatory cytokines. Longmu Piyandu prescription can improve AD-like skin lesions in BALB/c mice by promoting ROS clearance， enhancing TAOC， and inhibiting oxidative stress， thus protecting the skin barrier and reducing inflammation.

Objective:To evaluate the effect of Kuntai capsules combined with progestin-primed ovarian stimulation (PPOS) protocol on the ovarian response, laboratory parameters and embryo transfer outcomes of expected poor ovarian response (POR) patients undergoing invitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods:A retrospective cohort study was performed at Center for Reproductive Medicine, Jiangxi Maternal and Child Health Hospital from June 2020 to July 2023. A total of 1 733 expected POR patients were enrolled and divided into the PPOS only group (control group) and the Kuntai capsules combined with PPOS regimen group (trial group). After a 1∶1 propensity score matching, 423 cases were included in each group. Further analyses and comparisons were made between the two groups, concerning the ovarian stimulation outcomes, embryo laboratory parameters and pregnancy rates after the first frozen-thawed embryo transfer cycles. The primary outcome measure was the number of oocytes retrieved.Results:No statistically significant difference was found in baseline characteristics after matching between the two groups (all P>0.05). Compared with control group, the number of oocytes retrieved did not differ significantly in the trial group ( P=0.295). The level of luteinizing hormone (LH) on the trigger day [3.3 (2.2, 5.0) U/L], the number of available blastocysts formed [1 (0, 1)] and the available blastocyst formation rate [46.2% (85/184)] in the trial group were significantly higher than those in control group [2.9 (1.9, 4.5) U/L, P=0.004; 0 (0, 1), P=0.034; 30.0% (48/160), P=0.002]. However, there were no significant differences in the duration and dosage of gonadotropin used, progesterone and estradiol levels on the trigger day, incidence of premature LH surge, number/rate of fertilized oocytes, number/rate of cleaved embryos, number/rate of high-quality embryos, as well as rate of unavailable embryos (all P>0.05). Compared with control group, the trial group demonstrated comparable implantation rate, clinical pregnancy rate, early miscarriage rate, ongoing pregnancy rate and live birth rate after the first frozen-thawed embryo transfer cycles (all P>0.05). Conclusion:Compared with the PPOS only regimen, the combination of Kuntai capsules and PPOS did not increase the number of oocytes retrieved but could increase the number and rate of available blastocysts formed in expected POR patients.

Objective:To explore the clinical application value of third-generation sequencing technology in preimplantation genetic testing (PGT) of an ATR-X syndrome pedigree with chromosomal microduplication.Methods:The study selected a pedigree with a suspected ATR-X syndrome child at Assisted Reproductive Center of Jiangxi Maternal and Child Health Hospital in October 2022. After chromosome copy number variation sequencing (CNV Seq) detection, it was found that the female carried a 550 kb heterozygous microrepeat with unclear clinical significance in the Xq21.1 region, which involved partial sequences of the ATRX gene. The third generation long read sequencing technology was used to detect the female genome sequence, determine the physical location of the insertion of the above repeat into the genome, clarify the pathogenicity of the repeat, and obtain a single nucleotide polymorphism (SNP) haplotype linked to the above micro repeat inheritance. After the couple's full informed consent, PGT was performed to assist pregnancy. One haploid embryo without pathogenic microduplication was selected for transfer. To verify the consistency with PGT test results, amniocentesis prenatal diagnosis was performed in the second trimester after successful pregnancy, and the fetus was followed up after birth. Results:The results of the third generation long read sequencing and Sanger sequencing verification showed that the Xq21.1 microrepeat carried by the female was inserted into the genome chrX: 76804463-76804464 (GRCh37/hg19), which is an intra tandem repeat of the ATRX gene, and it is predicted that it may cause damage to the normal function of the ATRX protein. After one cycle of PGT treatment, 27 oocytes were obtained and 13 blastocysts were successfully developed after intracytoplasmic sperm injection (ICSI). Through genetic testing, it was found that two blastocysts were haploid embryos without carrying the aforementioned pathogenic microduplication. After thawing and transferring one of the blastocysts, the pregnancy was achieved, and the prenatal diagnosis results of amniocentesis in the second trimester were consistent with the PGT results. In November 2023, at 39 +5 weeks of pregnancy, a female live baby was delivered by natural delivery, and she is in good health. Conclusion:The third-generation sequencing technology has significant advantages in PGT detection of clinically ambiguous microreplicates with functional deficiency due to its long read length characteristics. It can not only determine the location of microreplicates inserted into the genome and determine their pathogenicity, but also obtain SNP haplotypes that are linked to the target mutation, thus preparing for subsequent embryo detection.

Objective:To evaluate the effect of Kuntai capsules combined with progestin-primed ovarian stimulation (PPOS) protocol on the ovarian response, laboratory parameters and embryo transfer outcomes of expected poor ovarian response (POR) patients undergoing invitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods:A retrospective cohort study was performed at Center for Reproductive Medicine, Jiangxi Maternal and Child Health Hospital from June 2020 to July 2023. A total of 1 733 expected POR patients were enrolled and divided into the PPOS only group (control group) and the Kuntai capsules combined with PPOS regimen group (trial group). After a 1∶1 propensity score matching, 423 cases were included in each group. Further analyses and comparisons were made between the two groups, concerning the ovarian stimulation outcomes, embryo laboratory parameters and pregnancy rates after the first frozen-thawed embryo transfer cycles. The primary outcome measure was the number of oocytes retrieved.Results:No statistically significant difference was found in baseline characteristics after matching between the two groups (all P>0.05). Compared with control group, the number of oocytes retrieved did not differ significantly in the trial group ( P=0.295). The level of luteinizing hormone (LH) on the trigger day [3.3 (2.2, 5.0) U/L], the number of available blastocysts formed [1 (0, 1)] and the available blastocyst formation rate [46.2% (85/184)] in the trial group were significantly higher than those in control group [2.9 (1.9, 4.5) U/L, P=0.004; 0 (0, 1), P=0.034; 30.0% (48/160), P=0.002]. However, there were no significant differences in the duration and dosage of gonadotropin used, progesterone and estradiol levels on the trigger day, incidence of premature LH surge, number/rate of fertilized oocytes, number/rate of cleaved embryos, number/rate of high-quality embryos, as well as rate of unavailable embryos (all P>0.05). Compared with control group, the trial group demonstrated comparable implantation rate, clinical pregnancy rate, early miscarriage rate, ongoing pregnancy rate and live birth rate after the first frozen-thawed embryo transfer cycles (all P>0.05). Conclusion:Compared with the PPOS only regimen, the combination of Kuntai capsules and PPOS did not increase the number of oocytes retrieved but could increase the number and rate of available blastocysts formed in expected POR patients.

Objective:To explore the clinical application value of third-generation sequencing technology in preimplantation genetic testing (PGT) of an ATR-X syndrome pedigree with chromosomal microduplication.Methods:The study selected a pedigree with a suspected ATR-X syndrome child at Assisted Reproductive Center of Jiangxi Maternal and Child Health Hospital in October 2022. After chromosome copy number variation sequencing (CNV Seq) detection, it was found that the female carried a 550 kb heterozygous microrepeat with unclear clinical significance in the Xq21.1 region, which involved partial sequences of the ATRX gene. The third generation long read sequencing technology was used to detect the female genome sequence, determine the physical location of the insertion of the above repeat into the genome, clarify the pathogenicity of the repeat, and obtain a single nucleotide polymorphism (SNP) haplotype linked to the above micro repeat inheritance. After the couple's full informed consent, PGT was performed to assist pregnancy. One haploid embryo without pathogenic microduplication was selected for transfer. To verify the consistency with PGT test results, amniocentesis prenatal diagnosis was performed in the second trimester after successful pregnancy, and the fetus was followed up after birth. Results:The results of the third generation long read sequencing and Sanger sequencing verification showed that the Xq21.1 microrepeat carried by the female was inserted into the genome chrX: 76804463-76804464 (GRCh37/hg19), which is an intra tandem repeat of the ATRX gene, and it is predicted that it may cause damage to the normal function of the ATRX protein. After one cycle of PGT treatment, 27 oocytes were obtained and 13 blastocysts were successfully developed after intracytoplasmic sperm injection (ICSI). Through genetic testing, it was found that two blastocysts were haploid embryos without carrying the aforementioned pathogenic microduplication. After thawing and transferring one of the blastocysts, the pregnancy was achieved, and the prenatal diagnosis results of amniocentesis in the second trimester were consistent with the PGT results. In November 2023, at 39 +5 weeks of pregnancy, a female live baby was delivered by natural delivery, and she is in good health. Conclusion:The third-generation sequencing technology has significant advantages in PGT detection of clinically ambiguous microreplicates with functional deficiency due to its long read length characteristics. It can not only determine the location of microreplicates inserted into the genome and determine their pathogenicity, but also obtain SNP haplotypes that are linked to the target mutation, thus preparing for subsequent embryo detection.

Objective:To explore eye movement characteristics in newly diagnosed, drug-naive Parkinson′s disease (PD) patients and their correlation with motor and non-motor symptoms.Methods:Seventy-five newly diagnosed, drug-naive PD patients and 46 healthy controls (HCs) were included in this cross-sectional study. Patients were recruited from the Department of Neurology, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine from November 2017 to December 2021, while HCs were recruited from the local community during the same period. For PD patients, motor severity was measured with the modified Hoehn and Yahr stage, Movement Disorder Society Unified Parkinson′s Disease Rating Scale part Ⅲ and the Freezing of Gait questionnaire. Non-motor symptoms were evaluated by serial scales such as Non-Motor Symptoms Questionnaire, 16-item odor identification test from Sniffin Sticks, 17-item Hamilton Rating Scale for Depression, Chinese version of Mini-Mental State Examination, Montreal Cognitive Assessment Basic and REM Behavior Disorder Screening Questionnaire. All subjects underwent oculomotor test including pro-saccade task and smooth pursuit eye movement (SPEM) task in the horizontal direction via videonystagmography. Visually guided saccade latency, saccadic accuracy and gain in SPEM at three frequencies (0.1, 0.2, 0.4 Hz) of the horizontal axis were compared between the 2 groups. The association between key oculomotor parameters and clinical phenotypes was explored in PD patients. The receiver operating characteristic (ROC) analyses of eye movement parameters as independent factors were also performed for detecting PD from HCs, then combining the saccadic latency, saccadic accuracy and the most significant SPEM gain (0.4 Hz) as the model to distinguish PD from HCs.Results:Relative to HCs, newly diagnosed, drug-naive PD patients showed prolonged saccadic latency [(210.4±41.3) ms vs (191.3±18.9) ms, t=-3.445, P=0.001] and decreased saccadic accuracy (88.4%±6.8% vs 92.2%±6.1%, t=3.064, P=0.003). SPEM gain in PD was uniformly reduced at each frequency(0.1 Hz: 0.68±0.15 vs 0.74±0.14, t=2.261, P=0.026; 0.2 Hz: 0.72±0.16 vs 0.79±0.16, t=2.704, P=0.008; 0.4 Hz: 0.67±0.19 vs 0.78±0.19, t=2.937, P=0.004). The ROC analyses of saccade latency, saccadic accuracy and gain in SPEM at 0.1, 0.2, 0.4 Hz as independent factors for detecting PD from HCs showed that the area under the curve (AUC) of each parameter was lower than 0.7: the AUC of saccade latency was 0.641 ( P=0.010), the AUC of saccadic accuracy was 0.681 ( P=0.001), the AUC of gain in SPEM at 0.1 Hz was 0.616 ( P=0.032), at 0.2 Hz was 0.652 ( P=0.005), at 0.4 Hz was 0.660 ( P=0.003). Combining the saccadic latency, saccadic accuracy and the most significant SPEM gain (0.4 Hz) revealed that the model could significantly distinguish PD from HCs with an 80.4% sensitivity and a 73.3% specificity (AUC=0.780, P<0.001). Prolonged saccadic latency was correlated with long disease duration ( β=0.334, 95% CI 0.014-0.654, P=0.041), whereas decreased SPEM gain was associated with severe motor symptoms in newly diagnosed drug-naive PD patients (0.1 Hz: β=-0.004, 95% CI -0.008--0.001, P=0.036; 0.4 Hz: β=-0.006, 95% CI -0.011--0.001, P=0.012). Conclusions:Ocular movements are impaired in newly diagnosed, drug-naive PD patients. These changes could be indicators for disease progression in PD.

Objective:To analyze the clinical practice of fertility preservation in unmarried young female breast cancer patients in China.Methods:This was a retrospective study of 13 unmarried female patients who underwent fertility preservation due to breast cancer in the Reproductive Medicine Center of International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong University School of Medicine from January 2019 to December 2020.Results:The age of the patients was (31.00±3.98) years old. Nine patients used follicular phase letrozole (LE)+gonadotropin (Gn) ovarian stimulation protocol, 3 patients used luteal phase LE+Gn stimulation protocol, and 1 patient used natural cycle protocol. Eleven patients completed the fertility preservation procedure, with a total Gn used dosage of (1 865.91±501.03) U, estradiol level at trigger of (1 223.85±709.75) ng/L, and 9.91±4.41 frozen oocytes. All of them used vitrification freezing.Conclusion:Oocyte vitrification frozen after LE+Gn ovarian stimulation is the preferred solution in young unmarried breast cancer patients for fertility preservation. However, promoting the awareness of fertility preservation, improving communication between clinical departments, and increasing clinical practice still need to be promoted.

Objective:To analyze the clinical practice of fertility preservation in unmarried young female breast cancer patients in China.Methods:This was a retrospective study of 13 unmarried female patients who underwent fertility preservation due to breast cancer in the Reproductive Medicine Center of International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong University School of Medicine from January 2019 to December 2020.Results:The age of the patients was (31.00±3.98) years old. Nine patients used follicular phase letrozole (LE)+gonadotropin (Gn) ovarian stimulation protocol, 3 patients used luteal phase LE+Gn stimulation protocol, and 1 patient used natural cycle protocol. Eleven patients completed the fertility preservation procedure, with a total Gn used dosage of (1 865.91±501.03) U, estradiol level at trigger of (1 223.85±709.75) ng/L, and 9.91±4.41 frozen oocytes. All of them used vitrification freezing.Conclusion:Oocyte vitrification frozen after LE+Gn ovarian stimulation is the preferred solution in young unmarried breast cancer patients for fertility preservation. However, promoting the awareness of fertility preservation, improving communication between clinical departments, and increasing clinical practice still need to be promoted.

Objective To establish an evaluation method of uncertainty for the determination of lead in ambient air PM_2.5 by inductively coupled plasma mass spectrometry (ICP-MS). Methods According to HJ657-2013 ^“Determination of Lead and Other Metal Elements in Air and Exhaust Particulates by Inductively Coupled Plasma Mass Spectrometry^”, the concentration of lead in ambient air PM2.5 was determined. A mathematical model was established to analyze the source of uncertainty and to calculate each component of uncertainty. The components were combined into the standard uncertainty, and the relative expanded uncertainty was finally obtained. Results The expanded uncertainty of lead in ambient air PM2.5 was 0.16 ng/m³, and the measurement result of lead was (1.75±0.16)ng/m³. Conclusion The main source of uncertainty of this method comes from the relative standard uncertainty introduced by sample pretreatment.