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 observe the clinical efficacy of Fangxiang Wentong Powder(composed of Salviae Miltiorrhizae Radix et Rhizoma,Chuanxiong Rhizoma,Alpiniae Officinarum Rhizoma,Piper Longum,and Corydalis Rhizoma)acupoint application combined with Kuanxiong Aerosol in treating female patients with coronary slow flow(CSF)associated angina.Methods After sample size estimation,119 female inpatients diagnosed as CSF associated angina and differentiated as chest-qi obstruction with yang deficiency and cold accumulation syndrome of traditional Chinese medicine(TCM)were collected from the cardiovascular departments of the Third People's Hospital of Fujian University of Traditional Chinese Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Chinese Medicine),Zhanjiang First Traditional Chinese Medicine Hospital,and Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2023 to March 2024.Using a table of random numbers,the patients were divided into a treatment group(84 cases)and a control group(35 cases)in a ratio of 5∶2.The control group was treated with Isosorbide Monomitrate Sustained-Release Capsules,while the treatment group was treated with application of Fangxiang Wentong Powder on acupoints of Danzhong(CV17),Gaohuang(BL43),and Xinshu(BL15),together with sublingual administration of Kuanxiong Aerosol.The treatment course for both groups covered 7 days.Before and after treatment,the changes in the simplified Seattle Angina Questionnaire(SAQ-7)scores,Chinese Quality of Life Questionnaire for Cardiovascular Patients(CQQC)scores,6-minute walking distance(6MWD),serum C-reactive protein(CRP)level,and white blood cell-to-mean platelet volume ratio(WMR)were observed.The TCM symptom efficacy was compared between the two groups,and adverse reactions were monitored.Results(1)There were 8 patients withdrew in the treatment group for failing in completing the questionnaires,and 2 patients withdrew in the control group for suffering headaches after taking nitrates.Eventually,109 patients completed the trial,including 76 in the treatment group and 33 in the control group.(2)After 7 days of treatment,the total effective rate for TCM symptom efficacy in the treatment group was 86.84%(66/76),and that in the control group was 72.73%(24/33).The intergroup comparison(tested by the chi-square test)showed that the TCM symptom efficacy in the treatment group was significantly superior to that in the control group.(3)After treatment,both groups showed improvements in the scores of activity limitation and angina frequency items of SAQ-7,and the treatment group also showed improvements in the scores of subjective satisfaction item of SAQ-7(P＜0.05).The treatment group's improvements in the scores of activity limitation,angina frequency,and subjective satisfaction items of SAQ-7 were significantly superior to those of the control group(P＜0.05).(4)After treatment,the CQQC scores in the treatment group was significantly improved(P＜0.05),while no significant improvement was observed in the control group(P＞0.05).The improvement of CQQC scores in the treatment group was significantly superior to that in the control group(P＜0.05).(5)After treatment,both groups showed improvements in 6MWD(P＜0.05),and the improvement in the treatment group was significantly superior to that in the control group(P＜0.05).(6)After treatment,the serum levels of inflammatory indicators of CRP and WMR in the treatment group were significantly improved(P＜0.05),while no significant improvements were presented in the control group(P＞0.05).The improvements in serum CRP level and WMR value in the treatment group were significantly superior to those in the control group(P＜0.05).(7)No adverse reactions were found in the treatment group,indicating high safety.Conclusion Fangxiang Wentong Powder acupoint application combined with Kuanxiong Aerosol is more effective than Isosorbide Monomitrate Sustained-Release Capsules,improving exercise tolerance,decreasing inflammatory factor levels,and improving the quality of life to some extent during the treatment of female patients with CSF.

Objective: To study the impact of disaggregation on the quality of in vitro aggregated apheresis platelets. Methods: The apheresis platelets collected from Guangzhou Blood Center served as the study samples. The in vitro aggregated apheresis platelets after successful disaggregation were designated as the experimental group (referred to as the aggregation group), and apheresis platelets without in vitro aggregation during collection served as the control group. The product volume, platelet content, residual white blood cells, red blood cell contamination, pH value, CD62p expression rate, platelet morphology score and thromboelastography of both groups were respectively detected. Results: The routine quality control indicators such as product volume, platelet content, residual white blood cells, red blood cell contamination, and pH value of both groups all met the quality requirements. There were statistically significant differences in pH value (7.180 vs 7.071) between the two groups (P<0.05). There was no significant difference in product volume, platelet content, residual white blood cells, and red blood cell contamination between the two groups (P>0.05). The CD62p expression rate of the aggregation group was higher than that of the control group (42.386% vs 17.310%, P<0.05), while the cell morphology score of the aggregation group was lower than that of the control group (132.066 vs 141.166, P<0.05). No statistically significant differences were found in thromboelastography parameters (R-CK, K-CK, α angle, MA-CK, CI-CK) between the two groups (P>0.05). Conclusion: After the disaggregation of in vitro-aggregated apheresis platelets, the quality indicators met the national quality requirements. Although the expression rate of CD62p increased and the cell morphology score decreased, there were no statistically significant differences in the thromboelastography parameters between the two groups. This indicates that although some platelet activation occurred, it did not affect the hemostatic function of the platelets.

Objective To investigate the suitable habitats of Phlebotomus chinensis in Gansu Province, so as provide insights into effective management of mountain-type zoonotic visceral leishmaniasis (MT-ZVL). Methods The geographical coordinates of locations where MT-ZVL cases were reported were retrieved in Gansu Province from 2015 to 2023, and data pertaining to 26 environmental variables were captured, including 19 climatic variables (annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the coldest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter), five geographical variables (elevation, annual normalized difference vegetation index, vegetation type, landform type and land use type), and two population and economic variables (population distribution and gross domestic product). Twelve species distribution models were built using the biomod2 package in R project, including surface range envelope (SRE) model, generalized linear model (GLM), generalized additive model (GAM), multivariate adaptive regression splines (MARS) model, generalized boosted model (GBM), classification tree analysis (CTA) model, flexible discriminant analysis (FDA) model, maximum entropy (MaxEnt) model, optimized maximum entropy (MAXNET) model, artificial neural network (ANN) model, random forest (RF) model, and extreme gradient boosting (XGBOOST) model. The performance of 12 models was evaluated using the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS), and Kappa coefficient, and single models with high performance was selected to build the optimal ensemble models. Factors affecting the survival of Ph. chinensis were identified based on climatic, geographical, population and economic variables. In addition, the suitable distribution areas of Ph. chinensis were predicted in Gansu Province under shared socioeconomic pathway 126 (SSP126), SSP370 and SSP585 scenarios based on climatic data during the period from 1991 to 2020, from 2041 to 2060 (2050s), and from 2081 to 2100 (2090s) . Results A total of 11 species distribution models were successfully built for prediction of potential distribution areas of Ph. chinensis in Gansu Province, and the RF model had the highest predictive accuracy (AUC = 0.998). The ensemble model built based on the RF model, XGBOOST model, GLM, and MARS model had an increased predictive accuracy (AUC = 0.999) relative to single models. Among the 26 environmental factors, precipitation of the wettest quarter (12.00%), maximum temperature of the warmest month (11.58%), and annual normalized difference vegetation index (11.29%) had the greatest contributions to suitable habitats distribution of Ph. sinensis. Under the climatic conditions from 1991 to 2020, the potential suitable habitat area for Ph. chinensis in Gansu Province was approximately 5.80 × 10⁴ km², of which the highly suitable area was 1.42 × 10⁴ km², and primarily concentrated in the southernmost region of Gansu Province. By the 2050s, the unsuitable and lowly suitable areas for Ph. chinensis in Gansu Province had decreased by varying degrees compared to that of 1991 to 2020 period, while the moderately and highly suitable areas exhibited expansion and migration. By the 2090s, under the SSP126 scenario, the suitable habitat area for Ph. chinensis increased significantly, and under the SSP585 scenario, the highly suitable areas transformed into extremely suitable areas, also showing substantial growth. Future global warming is conducive to the survival and reproduction of Ph. chinensis. From the 2050s to the 2090s, the highly suitable areas for Ph. chinensis in Gansu Province will be projected to expand northward. Under the SSP126 scenario, the suitable habitat area for Ph. chinensis in Gansu Province is expected to increase by 194.75% and 204.79% in the 2050s and 2090s, respectively, compared to that of the 1991 to 2020 period. Under the SSP370 scenario, the moderately and highly suitable areas will be projected to increase by 164.40% and 209.03% in the 2050s and 2090s, respectively, while under the SSP585 scenario, they are expected to increase by 195.98% and 211.66%, respectively. Conclusions The distribution of potential suitable habitats of Ph. sinensis gradually shifts with climatic changes. Intensified surveillance and management of Ph. sinensis is recommended in central and eastern parts of Gansu Province to support early warning of MT-ZVL.

Objective To analyze the drug resistance and genomic characteristics of clinically isolated hypermuco-viscous Klebsiella pneumoniae(hmKp).Methods Klebsiella pneumoniae isolated from clinical specimens from the sentinel hospitals of National Pathogen Identification Network in Huai'an City from 2019 to 2023 were collected,strains were identified by VITEK 2 Compact,mucus phenotype was determined by string test,resistance of hmKp was determined by microbroth dilution method.Molecular typing was conducted by whole-genome sequencing tech-nology,annotation of virulence and resistance genes carried by strains was performed.Results A total of 60 strains of hmKp were collected,mainly isolated from sputum specimens(33.33％)and blood specimens(28.33％).The average genome size of 60 strains was 5.6 Mb,with an average GC content of 57.09％.Multilocus sequence typing(MLST)showed that there were 28 ST types,with the dominant ST types being ST1 1(11.67％),ST15(11.67％),and ST412(10.00％).Core genome MLST(cgMLST)analysis revealed that some strains were highly homologous,and no outbreak strains were found.Multidrug-resistant strains accounted for 60.00％,while imipe-nem-resistant strains accounted for 28.33％.64 types of resistance genes were carried,84.38％of which were loca-ted on mobile element.The carriage rates of fosfomycin-related resistance genes and extended-spectrum β-lactam an-tibiotic-related resistance genes were relatively high in the strains,at 100％and 98.33％,respectively.Among car-bapenem-resistance gene blaKPC-2,21.67％was carried by ST11,ST1,and ST15 strains,among bla NDM-5 gene,3.33％was carried by ST76 strain.A total of 101 virulence genes were carried,most were Colibactin and type Ⅵsecretion system-related virulence genes.All strains carried capsule synthesis regulation-related genes(rcsA,rcsB),efflux pump-related genes(acrA,acrB),and enterobacterin-related genes(entABCEF,fepABCDFG,fes).Conclusion Clinically isolated hmKp in Huai'an exhibits multidrug resistance,with dominant ST types ST11,ST15,and ST412,carrying multiple horizontally transferable resistance and virulence genes,prevention and control measures should be strengthened.

Agenesis of the dorsal pancreas(ADP)is an extremely rare congenital pancreatic malformation characterized by the absence or hypoplasia of the pancreatic body and tail.Its pathogenesis is closely related to abnormal embryonic development of the ventral and dorsal pancreatic buds,governed by a complex network of transcription factors,including HLXB9,HNF1B,PDX1,PTF1A,GATA4,and GATA6.The clinical spectrum of ADP is highly variable,ranging from asymptomatic cases to manifestations such as abdominal pain,diabetes mellitus,or pancreatitis.Imaging modalities-including ultrasonography,CT,magnetic resonance cholangiopancreatography,and endoscopic retrograde cholangio-pancreatography-serve as the main diagnostic tools,with characteristic findings of absent pancreatic body and tail accompanied by compensatory enlargement of the pancreatic head.ADP is frequently associated with congenital anomalies of the kidney,biliary tract,cardiovascular system,or genital organs.Management is primarily symptomatic,with insulin replacement for diabetes and pancreatic enzyme supplementation for exocrine insufficiency.Advances in genetic sequencing and stem cell research have deepened understanding of the pathogenesis,genetic background,and potential therapeutic strategies of ADP.This review summarizes current progress in embryology,genetics,clinical features,diagnosis,and treatment of ADP,aiming to improve clinical recognition and guide future investigations.

Agenesis of the dorsal pancreas(ADP)is an extremely rare congenital pancreatic malformation characterized by the absence or hypoplasia of the pancreatic body and tail.Its pathogenesis is closely related to abnormal embryonic development of the ventral and dorsal pancreatic buds,governed by a complex network of transcription factors,including HLXB9,HNF1B,PDX1,PTF1A,GATA4,and GATA6.The clinical spectrum of ADP is highly variable,ranging from asymptomatic cases to manifestations such as abdominal pain,diabetes mellitus,or pancreatitis.Imaging modalities-including ultrasonography,CT,magnetic resonance cholangiopancreatography,and endoscopic retrograde cholangio-pancreatography-serve as the main diagnostic tools,with characteristic findings of absent pancreatic body and tail accompanied by compensatory enlargement of the pancreatic head.ADP is frequently associated with congenital anomalies of the kidney,biliary tract,cardiovascular system,or genital organs.Management is primarily symptomatic,with insulin replacement for diabetes and pancreatic enzyme supplementation for exocrine insufficiency.Advances in genetic sequencing and stem cell research have deepened understanding of the pathogenesis,genetic background,and potential therapeutic strategies of ADP.This review summarizes current progress in embryology,genetics,clinical features,diagnosis,and treatment of ADP,aiming to improve clinical recognition and guide future investigations.

Objective To analyze the drug resistance and genomic characteristics of clinically isolated hypermuco-viscous Klebsiella pneumoniae(hmKp).Methods Klebsiella pneumoniae isolated from clinical specimens from the sentinel hospitals of National Pathogen Identification Network in Huai'an City from 2019 to 2023 were collected,strains were identified by VITEK 2 Compact,mucus phenotype was determined by string test,resistance of hmKp was determined by microbroth dilution method.Molecular typing was conducted by whole-genome sequencing tech-nology,annotation of virulence and resistance genes carried by strains was performed.Results A total of 60 strains of hmKp were collected,mainly isolated from sputum specimens(33.33％)and blood specimens(28.33％).The average genome size of 60 strains was 5.6 Mb,with an average GC content of 57.09％.Multilocus sequence typing(MLST)showed that there were 28 ST types,with the dominant ST types being ST1 1(11.67％),ST15(11.67％),and ST412(10.00％).Core genome MLST(cgMLST)analysis revealed that some strains were highly homologous,and no outbreak strains were found.Multidrug-resistant strains accounted for 60.00％,while imipe-nem-resistant strains accounted for 28.33％.64 types of resistance genes were carried,84.38％of which were loca-ted on mobile element.The carriage rates of fosfomycin-related resistance genes and extended-spectrum β-lactam an-tibiotic-related resistance genes were relatively high in the strains,at 100％and 98.33％,respectively.Among car-bapenem-resistance gene blaKPC-2,21.67％was carried by ST11,ST1,and ST15 strains,among bla NDM-5 gene,3.33％was carried by ST76 strain.A total of 101 virulence genes were carried,most were Colibactin and type Ⅵsecretion system-related virulence genes.All strains carried capsule synthesis regulation-related genes(rcsA,rcsB),efflux pump-related genes(acrA,acrB),and enterobacterin-related genes(entABCEF,fepABCDFG,fes).Conclusion Clinically isolated hmKp in Huai'an exhibits multidrug resistance,with dominant ST types ST11,ST15,and ST412,carrying multiple horizontally transferable resistance and virulence genes,prevention and control measures should be strengthened.