Objective:To assess the feasibility of first polar body transfer (PB1T) combined with preimplantation mitochondrial genetic testing for blocking the transmission of a pathogenic mitochondrial DNA 8993T>G mutation.Methods:A Chinese family affected with Leigh syndrome which had attended the Reproductive Medicine Centre of the First Affiliated Hospital of Anhui Medical University in September 2021 was selected as the study subject. Controlled ovarian hyperstimulation was carried out for the proband after completing the detection of the mitochondrial DNA 8993T>G mutation load among the pedigree members. Mature MII oocytes were inseminated by intracytoplasmic sperm injection (ICSI), cultured in vitro for 5 to 6 days to the blastocyst stage, and trophoblastocytes were obtained by microbiopsy. Mitochondrial DNA testing (PGT-MT) and chromosomal aneuploidy (PGT-A) analyses were carried out after whole-genome amplification, and the embryos with zero mutation load were selected for transfer. Amniotic fluid and umbilical cord blood samples were collected during middle pregnancy and after birth respectively for mitochondrial DNA testing to verify the reliability of embryo screening. As an attempt, PB1 with good morphology of MⅡ oocytes was selected for transfer into the enucleated oocytoplasm from healthy donors, followed by ICSI fertilization, blastocyst culture and PGT of embryos using the same procedure. This study has been approved by the Ethics Committee of the First Affiliated Hospital of Anhui Medical University (No. 2021zhyx-B12). Results:An antagonist protocol was used for ovarian stimulation, and a total of 19 oocytes were obtained, of which 14 MⅡ were fertilized by ICSI, and 2 had developed into blastocysts. PGT-MT was carried out on biopsied trophoblastocytes, in which the mitochondrial DNA 8993T>G mutation load was not detected in one embryo, the other was 100% mutated, and the mutation loads of the remaining unfertilized eggs and developmentally arrested embryos ranged from 0% ~ 100%, presenting a clear biased distribution. With fully informed consent, one PGT-MT zero mutation load blastocyst was transferred and clinical pregnancy was achieved. Mitochondrial DNA and chromosomal testing of amniotic fluid cells during middle pregnancy had revealed no abnormalities. The proband had delivered a healthy boy through Caesarean section at 39+ 5 weeks of gestation, and no mutation was detected in the cord blood sample. Five well-formed PBs from 14 eggs were selected for PB1 transfer, followed by ICSI and culture, and two of the reconstituted embryos had formed blastocysts, with none of the above mutations detected in the biopsied samples.Conclusion:The PGT-MT technology can help families affected with mitochondrial diseases to have healthy offspring. PB1 transfer in combination with ICSI and PGT-MT holds the promise of turning waste into treasure and providing an alternative means of fertility for such families.

OBJECTIVE: To assess the feasibility of first polar body transfer (PB1T) combined with preimplantation mitochondrial genetic testing for blocking the transmission of a pathogenic mitochondrial DNA 8993T>G mutation. METHODS: A Chinese family affected with Leigh syndrome which had attended the Reproductive Medicine Centre of the First Affiliated Hospital of Anhui Medical University in September 2021 was selected as the study subject. Controlled ovarian hyperstimulation was carried out for the proband after completing the detection of the mitochondrial DNA 8993T>G mutation load among the pedigree members. Mature MII oocytes were inseminated by intracytoplasmic sperm injection (ICSI), cultured in vitro for 5 to 6 days to the blastocyst stage, and trophoblastocytes were obtained by microbiopsy. Mitochondrial DNA testing (PGT-MT) and chromosomal aneuploidy (PGT-A) analyses were carried out after whole-genome amplification, and the embryos with zero mutation load were selected for transfer. Amniotic fluid and umbilical cord blood samples were collected during middle pregnancy and after birth respectively for mitochondrial DNA testing to verify the reliability of embryo screening. As an attempt, PB1 with good morphology of MII oocytes was selected for transfer into the enucleated oocytoplasm from healthy donors, followed by ICSI fertilization, blastocyst culture and PGT of embryos using the same procedure. This study has been approved by the Ethics Committee of the First Affiliated Hospital of Anhui Medical University (No. 2021zhyx-B12). RESULTS: An antagonist protocol was used for ovarian stimulation, and a total of 19 oocytes were obtained, of which 14 MII were fertilized by ICSI, and 2 had developed into blastocysts. PGT-MT was carried out on biopsied trophoblastocytes, in which the mitochondrial DNA 8993T>G mutation load was not detected in one embryo, the other was 100% mutated, and the mutation loads of the remaining unfertilized eggs and developmentally arrested embryos ranged from 0% ~ 100%, presenting a clear biased distribution. With fully informed consent, one PGT-MT zero mutation load blastocyst was transferred and clinical pregnancy was achieved. Mitochondrial DNA and chromosomal testing of amniotic fluid cells during middle pregnancy had revealed no abnormalities. The proband had delivered a healthy boy through Caesarean section at 39⁺⁵ weeks of gestation, and no mutation was detected in the cord blood sample. Five well-formed PBs from 14 eggs were selected for PB1 transfer, followed by ICSI and culture, and two of the reconstituted embryos had formed blastocysts, with none of the above mutations detected in the biopsied samples. CONCLUSION The PGT-MT technology can help families affected with mitochondrial diseases to have healthy offspring. PB1 transfer in combination with ICSI and PGT-MT holds the promise of turning waste into treasure and providing an alternative means of fertility for such families.
Humans ; Preimplantation Diagnosis/methods* ; Female ; DNA, Mitochondrial/genetics* ; Genetic Testing/methods* ; Pregnancy ; Mitochondrial Diseases/genetics* ; Polar Bodies ; Adult ; Feasibility Studies ; Sperm Injections, Intracytoplasmic/methods* ; Embryo Transfer/methods* ; Mutation ; Male ; Blastocyst/metabolism* ; Pedigree

OBJECTIVE To get the current situation of healthcare associated infection(HAI)and implement targe-ted management through analyzing the trends of HAI incidence rate,the distribution of clinical departments,the sites of HAI and the composition of pathogens in a children's hospital in a five-year period.METHODS Data on HAI cases were collected from all inpatients in Children's Hospital of Soochow University from 2019 to 2023 through the real-time monitoring system of XINGLIN Healthcare acquired Infection Surveillance;Cochran-Armit-age test was used to check for the significant changes of distribution of HAI cases.RESULTS A total of 383,376 hospitalized patients were monitored from 2019 to 2023,of which HAI occurred on 6670 cases and 7209 case-times with an incidence rate of 1.74%and an incidence case rate of 1.88%.The top five departments of HAI inci-dence rates were cardiac,neonatal,surgical and pediatric intensive care unit and department of hematology.HAI mainly occurred in blood and alimentary system,upper and lower respiratory tract.A total of 2668 strains of path-ogenic bacteria were isolated,of which 1346 strains were gram-negative bacteria,1140 strains were gram-positive bacteria and 182 strains were fungi.The top three gram-negative bacteria were Klebsiella pneumoniae,Escherich-ia coli and Pseudomonas aeruginosa;the top three gram-negative pathogens were Streptococcus,Staphylococcus aureus and Staphylococcus epidermidis;and the top three fungi were Candida albicans,Candida parapsilosis and Candida tropicalis.CONCLUSIONS The HAI incidence rate of this hospital steadily declines in the past five years,however the same changes are not observed in the departments with high incidence of HAI.Attention should be paid to the raising bloodstream infections and detection rate of S.epidermidis.

Objective To explore the distribution characteristics and influencing factors of nasal colonized bacteria of healthcare workers(HCWs)in pediatric intensive care unit(PICU).Methods A cross-sectional study was con-ducted.Nasal swab specimens from 104 HCWs in the PICU of a hospital were collected for bacterial culture and an-timicrobial susceptibility testing.According to the identification and antimicrobial susceptibility testing results of strains,distribution characteristics of colonized bacteria was analyzed.Basic information of studied subjects were collected through questionnaire survey,and risk factors for colonized bacterial infection were conducted using logis-tic regression analysis.Results Among 104 specimens,colonized bacteria were detected from 66 specimens,with an overall detection rate of 63.46％.Gram-positive bacteria was mainly Staphylococcus aureus,with a detection rate of 34.62％(n=36),out of which methicillin-resistant Staphylococcus aureus(MRSA)accounted for 2.88％(n=3).Gram-negative bacteria was mainly Klebsiella spp.,with a detection rate of 21.15％(n=22).Multiva-riate logistic regression analysis showed that HCWs with junior professional titles(OR=11.400,95％CI:2.329-55.801,P=0.003)was an independent risk factor for Staphylococcus aureus colonization,and male(OR=4.260,95％CI:1.160-15.653,P=0.029)was an independent risk factor for Klebsiella spp.colonization.Conclusion Nasal cavity of HCWs in PICU has a high detection rate of colonized bacteria,with Staphylococcus aureus and Klebsiella spp.being the major colonized bacteria.

Objective To explore the distribution characteristics and influencing factors of nasal colonized bacteria of healthcare workers(HCWs)in pediatric intensive care unit(PICU).Methods A cross-sectional study was con-ducted.Nasal swab specimens from 104 HCWs in the PICU of a hospital were collected for bacterial culture and an-timicrobial susceptibility testing.According to the identification and antimicrobial susceptibility testing results of strains,distribution characteristics of colonized bacteria was analyzed.Basic information of studied subjects were collected through questionnaire survey,and risk factors for colonized bacterial infection were conducted using logis-tic regression analysis.Results Among 104 specimens,colonized bacteria were detected from 66 specimens,with an overall detection rate of 63.46％.Gram-positive bacteria was mainly Staphylococcus aureus,with a detection rate of 34.62％(n=36),out of which methicillin-resistant Staphylococcus aureus(MRSA)accounted for 2.88％(n=3).Gram-negative bacteria was mainly Klebsiella spp.,with a detection rate of 21.15％(n=22).Multiva-riate logistic regression analysis showed that HCWs with junior professional titles(OR=11.400,95％CI:2.329-55.801,P=0.003)was an independent risk factor for Staphylococcus aureus colonization,and male(OR=4.260,95％CI:1.160-15.653,P=0.029)was an independent risk factor for Klebsiella spp.colonization.Conclusion Nasal cavity of HCWs in PICU has a high detection rate of colonized bacteria,with Staphylococcus aureus and Klebsiella spp.being the major colonized bacteria.

OBJECTIVE To get the current situation of healthcare associated infection(HAI)and implement targe-ted management through analyzing the trends of HAI incidence rate,the distribution of clinical departments,the sites of HAI and the composition of pathogens in a children's hospital in a five-year period.METHODS Data on HAI cases were collected from all inpatients in Children's Hospital of Soochow University from 2019 to 2023 through the real-time monitoring system of XINGLIN Healthcare acquired Infection Surveillance;Cochran-Armit-age test was used to check for the significant changes of distribution of HAI cases.RESULTS A total of 383,376 hospitalized patients were monitored from 2019 to 2023,of which HAI occurred on 6670 cases and 7209 case-times with an incidence rate of 1.74%and an incidence case rate of 1.88%.The top five departments of HAI inci-dence rates were cardiac,neonatal,surgical and pediatric intensive care unit and department of hematology.HAI mainly occurred in blood and alimentary system,upper and lower respiratory tract.A total of 2668 strains of path-ogenic bacteria were isolated,of which 1346 strains were gram-negative bacteria,1140 strains were gram-positive bacteria and 182 strains were fungi.The top three gram-negative bacteria were Klebsiella pneumoniae,Escherich-ia coli and Pseudomonas aeruginosa;the top three gram-negative pathogens were Streptococcus,Staphylococcus aureus and Staphylococcus epidermidis;and the top three fungi were Candida albicans,Candida parapsilosis and Candida tropicalis.CONCLUSIONS The HAI incidence rate of this hospital steadily declines in the past five years,however the same changes are not observed in the departments with high incidence of HAI.Attention should be paid to the raising bloodstream infections and detection rate of S.epidermidis.

Objective:To assess the feasibility of first polar body transfer (PB1T) combined with preimplantation mitochondrial genetic testing for blocking the transmission of a pathogenic mitochondrial DNA 8993T>G mutation.Methods:A Chinese family affected with Leigh syndrome which had attended the Reproductive Medicine Centre of the First Affiliated Hospital of Anhui Medical University in September 2021 was selected as the study subject. Controlled ovarian hyperstimulation was carried out for the proband after completing the detection of the mitochondrial DNA 8993T>G mutation load among the pedigree members. Mature MII oocytes were inseminated by intracytoplasmic sperm injection (ICSI), cultured in vitro for 5 to 6 days to the blastocyst stage, and trophoblastocytes were obtained by microbiopsy. Mitochondrial DNA testing (PGT-MT) and chromosomal aneuploidy (PGT-A) analyses were carried out after whole-genome amplification, and the embryos with zero mutation load were selected for transfer. Amniotic fluid and umbilical cord blood samples were collected during middle pregnancy and after birth respectively for mitochondrial DNA testing to verify the reliability of embryo screening. As an attempt, PB1 with good morphology of MⅡ oocytes was selected for transfer into the enucleated oocytoplasm from healthy donors, followed by ICSI fertilization, blastocyst culture and PGT of embryos using the same procedure. This study has been approved by the Ethics Committee of the First Affiliated Hospital of Anhui Medical University (No. 2021zhyx-B12). Results:An antagonist protocol was used for ovarian stimulation, and a total of 19 oocytes were obtained, of which 14 MⅡ were fertilized by ICSI, and 2 had developed into blastocysts. PGT-MT was carried out on biopsied trophoblastocytes, in which the mitochondrial DNA 8993T>G mutation load was not detected in one embryo, the other was 100% mutated, and the mutation loads of the remaining unfertilized eggs and developmentally arrested embryos ranged from 0% ~ 100%, presenting a clear biased distribution. With fully informed consent, one PGT-MT zero mutation load blastocyst was transferred and clinical pregnancy was achieved. Mitochondrial DNA and chromosomal testing of amniotic fluid cells during middle pregnancy had revealed no abnormalities. The proband had delivered a healthy boy through Caesarean section at 39+ 5 weeks of gestation, and no mutation was detected in the cord blood sample. Five well-formed PBs from 14 eggs were selected for PB1 transfer, followed by ICSI and culture, and two of the reconstituted embryos had formed blastocysts, with none of the above mutations detected in the biopsied samples.Conclusion:The PGT-MT technology can help families affected with mitochondrial diseases to have healthy offspring. PB1 transfer in combination with ICSI and PGT-MT holds the promise of turning waste into treasure and providing an alternative means of fertility for such families.

Nucleotide-binding oligomerization domain 1 (NOD1), a cytosolic pattern recognition receptor protein, plays a crucial role in innate immune responses. However, the functional expression of NOD1 in mesenchymal stem cells (MSCs) derived from endometriosis remains unclear. The aim of this study was to explore the functions of NOD1 in ectopic endometrial lesions. Tissues and MSCs were isolated from both normal endometrium and endometriosis.Immunohistochemistry and real time quantitative polymerase chain reaction (RT-qPCR) were used to determine the expression of NOD1 in the tissues/MSCs. Quantification of various cytokines was performed using RT-qPCR and enzyme-linked immunosorbent assay. To confirm the proliferation, invasion/migration, and apoptotic viabilities of the samples, Cell Counting Kit-8, clonogenic formation, transwell assays, and apoptotic experiments were conducted.Higher levels of NOD1 expression were detected in the ectopic-MSCs obtained from endometriosis compared to those from the endometrium. The expression of interleukin-8 was higher in the ectopic-MSCs than in the eutopic-MSCs.Pretreatment with NOD1 agonist significantly enhanced the proliferation and invasion/migration of eutopic-MSCs.Additionally, the NOD1 inhibitor ML-130 significantly reduced the proliferation, clone formation, invasion, and migration abilities of the ectopic-MSCs, having no effect on their apoptosis capacity. Our findings suggest that the expression of NOD1 in ectopic-MSCs may contribute to the progression of ectopic endometrial lesions.

Objective To explore the application value of popaverine combined with stage rehabilitation for patients after free flap grafting for hand trauma.Methods Data of postoperative patients who underwent hand trauma from January 2021 to January 2022 in The 903rd Hospital of the Joint Logistics Support Force of the People's Liberation Army of Chinawere retrospectively collected,and they were divided into the combined rehabilitation group(poppadine combined with stage rehabilitation care)and the stage rehabilitation group(only stage rehabilitation care was applied)according to the treatment methods.Visual analog scale(VAS)scores,rehabilitation exercise adherence scores,postoperative rates of excellent cutaneous sensory function of the hand,total active mobility(TAM)of the fingers,Jamar grip strength,upper extremity functional assessment(DASH)scale scores,and the incidence of vascular crises were compared between the two groups.Results A total of 120 cases were included in the study,with 57 cases in the combined rehabilitation group and 63 cases in the stage rehabilitation group.There was no statistically significant difference in VAS scores,Jamar grip strength and DASH scores between the two groups before surgery(P＞0.05);at 14 days and 3 months of intervention,VAS scores of the two groups decreased compared with the pre-intervention period(P＜0.05),and VAS scores of the patients in the combined rehabilitation group were significantly lower than those in the stage rehabilitation group(P＜0.05).After 3 months of intervention,the rehabilitation exercise adherence score,the rate of excellent hand skin sensory function,and finger TAM of the patients in the combined rehabilitation group were higher than those in the stage rehabilitation group(P＜0.05);Jamar grip strength were elevated in both groups compared with the pre-intervention period,but the combined rehabilitation group were higher than those in the stage rehabilitation group(P＜0.05);DSAH scores were lower in both groups compared to pre-intervention,and were lower in the combined rehabilitation group than in the stage rehabilitation group;and during the 3 months of intervention,the incidence of vascular crisis was significantly lower in patients in the combined rehabilitation group than in the stage rehabilitation group(P＜0.05).Conclusion Opium poppy alkaloids combined with stage rehabilitation can effectively reduce patients'postoperative pain,enhance patients'adherence to rehabilitation exercises,effectively improve postoperative hand function and upper limb motor function,and reduce the incidence of vascular crisis.

Objective:To investigate the change of chemical exchange saturation transfer (CEST) imaging in the bilateral substantia nigra (SN) and red nucleus (RN) of Parkinson disease (PD), and to explore the value of CEST-MRI for the clinical application of PD.Methods:This was a cross-sectional study. A total of 45 PD patients (PD group) and 21 sex-, age-, and cognitive-function matched normal control subjects (NC group) were retrospectively enrolled from December 2012 to July 2015 in Beijing Hospital. All subjects underwent brain CEST-MRI and routine MRI. Based on the MATLAB software package, the 4-pool Lorentz fitting model was applied to analyze the signal change of CEST imaging, which could acquire the mean amplitudes of the 4-pool parameters including Amide, nuclear overhauser enhancement (NOE), direct water saturation (DS) and magnetization transfer (MT) in the bilateral SN and RN. Independent samples t test and Mann-Whitney U test were used to compare CEST parameters between the PD group and the NC group and controlled by Bonferroni correction. The combined model was constructed based on parameters with inter-group differences after correction. The receiver operating characteristic curve and area under the curve (AUC) were used to evaluate the diagnostic efficiency of the CEST parameters and the combined model. Results:Compared with the NC group, the left SN Amide value, left RN Amide value, and right SN NOE value were reduced in the PD group, and the difference was statistically significant ( t=-3.59, corrected P=0.026; t=-3.77, corrected P=0.016; Z=-3.27, corrected P=0.017). The left SN Amide value, the left RN Amide value, the right SN NOE value, and the combined model all had good diagnostic efficacy in the differentiation of the PD group from the HC group (AUCs of 0.78, 0.79, 0.75, and 0.81, respectively). The combined model had the highest AUC value (0.81) and specificity (97.78%), the Amide value of left SN had the highest sensitivity (93.33%). Conclusions:Quantitative analysis of CEST-MRI based on the 4-pool Lorentz fitting model shows significant differences in the CEST quantitative indicators of the SN and RN between the PD group and the NC group, demonstrating good potential for clinical application in the diagnosis of PD.