Objective:To determine the contents of four flavonoids by establishing HPLC specific chromatogram for Alpiniae Katsumadai Semen; To evaluate the differences of Alpiniae Katsumadai Semen from different producing areas.Methods:The specific chromatogram was developed on a column of Thermo Acclaim C18 with acetonitrile-0.1% phosphoric acid solution as the mobile phase by gradient elution at a flow rate of 1.0 ml/min. The detective wavelength was 260 nm, and the column temperature was 30 ℃. Similarity evaluation, PCA analysis, and OPLS-DA analysis were conducted. The contents of Alpinetin, Pinocembrin, Cardamonin, Alnustone in 16 batches of Alpiniae Katsumadai Semen.Results:There were 9 characteristic peaks in the specific chromatogram of Alpiniae Katsumadai Semen. Except the sample of S2 (Hainan producing area), the similarity of Alpiniae Katsumadai Semen in different producing areas was greater than 0.90; PCA analysis divided 16 batches of Alpiniae Katsumadai Semen into 2 categories, and OPLS-DA analysis identified 4 differential biomarkers, with the order of impact being peak 3>peak 5>Alpinetin>Cardamonin. Among them, the quality of Alpiniae Katsumadai Semen from Guangdong producing area was generally stable. Moreover, there were significant differences in the contents of Alpinetin and Cardamonin among the indicator components of Alpiniae Katsumadai Semen from different producing areas.Conclusion:This method can effectively analyze the differences in the quality of Alpiniae Katsumadai Semen from different producing areas, providing reference for the quality evaluation of Alpiniae Katsumadai Semen.

Intestinal stem cells(ISCs),locating at the base of intestinal crypts and pivotal in orchestrating the homeostasis and damage repair of the intestinal epithelium,are characterized by their self-renewal and multipotential differentiation capabilities.With the continuous discoveries of new ISCs and related markers,the ISC migration and regeneration model has been further improved,greatly promoting the research in related fields.Diet can regulate ISC glycolipid and energy metabolism through influencing the stem cell niche,subsequently modulating overall metabolism.This paper summarizes the biological features of both classical and newly discovered ISCs,and analyzes the effects of common nutrients and different dietary patterns on ISCs,hoping to provide insights for the precise nutrition prevention and treatment of chronic intestinal diseases.

Objective:To explore the effect of integrating open laboratories into Medical Microbiology experimental teaching for students majoring in Medical Laboratory Technology to enhance teaching quality.Methods:A total of 58 students from the grade 2020 majoring in Medical Laboratory Technology at a medical college were selected as the control group,receiving traditional teaching methods without open laboratory contents.A total of 58 students from the grade 2021 majoring in Medical Laboratory Technology were selected as the experimental group,integrating open laboratory contents into Medical Microbiology experimental teaching.The impact of open laboratories on teaching quality in Medical Microbiology experimental teaching was evaluated through analysis of final examination scores and questionnaire surveys.Results:Compared with the control group,the experimental group showed significant improvements in experimental report scores,experimental theory assessment scores,practical skill assessment scores,classroom performance scores,and overall scores(P＜0.01).The learning interest,multi-channel learning,mastery of key concepts,and ability to apply and expand knowledge of the experimental group also significantly improved(P＜0.01).The satisfaction rate of the experimental group in terms of participation level,acquired knowledge,mastered skills,and improved learning outcomes was all above 90%.Conclusion:Integrating open laboratory content into Medical Microbiology experimental teaching for students majoring in Medical Laboratory Technology can effectively stimulate students'engagement,enhance teaching quality,and help students better understand and master professional knowledge.

Objective To investigate the expression of the circular RNA circ-PHC3 in cervical cancer tissues and its regulatory mechanisms in the proliferation,migration,and invasion of cervical cancer cells.Methods The expression levels of circ-PHC3 in cervical cancer tissues and adjacent non-tumor tissues were analyzed using the GEO database.The correlation between circ-PHC3 expression and the clinical stage as well as prognosis of cervical cancer patients was also evaluated.The expression of circ-PHC3 in cervical cancer cell lines HCC94,C33A,HeLa,HCC1106,and SiHa was detected by real-time quantitative polymerase chain reaction(qRT-PCR).The cell line with the highest circ-PHC3 expression was selected for transfection with a circ-PHC3 inhibitor.The interaction between circ-PHC3 and miR-1179 was validated using a dual luciferase reporter gene assay.The expression levels of miR-1179 in transfected cells were further assessed by qRT-PCR.Functional assays,including colony formation,flow cytometry,wound healing,and Transwell assays,were conducted to evaluate cell proliferation,cell cycle progression,migration,and invasion,respectively.Western blot analysis was performed to determine the expression of key proteins associated with proliferation,migration,and invasion in circ-PHC3-modulated cells.Finally,in vivo experiments were carried out to investigate the impact of circ-PHC3 silencing on the growth and metastasis of cervical cancer cells in animal models.Results The expression level of circ-PHC3 in cervical cancer tissues was significantly higher than that in adjacent normal tissues(P<0.01).Furthermore,circ-PHC3 expression was significantly associated with the clinical stage of cervical cancer(P<0.01).Patients with high circ-PHC3 expression exhibited a notably lower survival rate compared to those with low circ-PHC3 expression(P<0.01).In cervical cancer cell lines including HCC94,C33A,HeLa,HCC1106,and SiHa,circ-PHC3 expression was markedly upregulated(all P<0.01),with the highest expression observed in HCC1106 cells(P<0.01).Circ-PHC3 was found to directly interact with miR-1179(P<0.01),and silencing circ-PHC3 significantly increased miR-1179 expression(P<0.01).Transfection of HCC1106 cells with a circ-PHC3 inhibitor significantly suppressed cell proliferation,migration,and invasion(all P<0.01),and induced cell cycle arrest(P<0.01);these effects were partially reversed by co-transfection with a miR-1179 inhibitor(all P<0.05).In HCC1106 cells with circ-PHC3 knockdown,the expression levels of key proteins associated with proliferation,migration,and invasion—Cyclin E,CDK2,MMP-9,and N-cadherin—were significantly reduced(all P<0.01),and this reduction was partially attenuated by miR-1179 inhibition(all P<0.01).In vivo experiments further demonstrated that circ-PHC3 knockdown significantly inhibited tumor growth and metastasis of HCC1106 cells(all P<0.01).Conclusions Circ-PHC3 is highly expressed in cervical cancer tissues,and its overexpression is significantly correlated with poor prognosis in patients with cervical cancer.Knockdown of circ-PHC3 upregulates the expression of miR-1179 and suppresses the proliferation,migration,and invasion of cervical cancer cells.

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 understand the species of mosquitoes and the status of important mosquito-borne viruses in the Ningxia surveyed regions, to identify the dominant mosquito species and virus types, and to analyze their genetic characteristics, providing a scientific basis for predicting and controlling mosquito-borne infectious diseases.Methods:Mosquitoes were collected using light traps in Yinchuan and Wuzhong cities of the Ningxia Hui Autonomous Region, and the collected mosquitoes were classified and identified. Real-time polymerase chain reaction (PCR) was used to detect the Japanese encephalitis virus (JEV), West Nile virus (WNV), Chikungunya virus (CHIKV), Sindbis virus (SINV) carried by mosquitoes. The positive sample was subjected to sequencing the whole genome, and the phylogenetic tree of virus strains was constructed using bioinformatics methods.Results:From June to August 2023, a total of 8 561 mosquitoes of 3 genera and 6 species were collected in Yinchuan and Wuzhong cities, Ningxia, among which Culex pipiens pallens was the dominant species with 3 050 individuals, accounting for 35.63%; Anopheles sinensis with 2 379 individuals, accounting for 27.79%; Culex tritaeniorhynchus with 1 489 individuals, accounting for 17.39%; Caspian Aedes with 1 468 individuals, accounting for 27.79%; Aedes vexans with 152 individuals, accounting for 1.78%; and Culex modestus with 23 individuals, accounting for 0.27%. JEV GIb type was detected in the specimens of Culex tritaeniorhynchus collected in Qingtongxia city. Conclusions:The dominant mosquito species in the surveyed areas of Ningxia are primarily Culex pipiens pallens, and JEV GIb virus was detected in Culex tritaeniorhynchus in Qingtongxia city. This study provides basic data for understanding the current status of mosquitoes and mosquito-borne viruses in the Ningxia region and offers scientific guidance for further public health prevention and control measures.

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 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 understand the species of mosquitoes and the status of important mosquito-borne viruses in the Ningxia surveyed regions, to identify the dominant mosquito species and virus types, and to analyze their genetic characteristics, providing a scientific basis for predicting and controlling mosquito-borne infectious diseases.Methods:Mosquitoes were collected using light traps in Yinchuan and Wuzhong cities of the Ningxia Hui Autonomous Region, and the collected mosquitoes were classified and identified. Real-time polymerase chain reaction (PCR) was used to detect the Japanese encephalitis virus (JEV), West Nile virus (WNV), Chikungunya virus (CHIKV), Sindbis virus (SINV) carried by mosquitoes. The positive sample was subjected to sequencing the whole genome, and the phylogenetic tree of virus strains was constructed using bioinformatics methods.Results:From June to August 2023, a total of 8 561 mosquitoes of 3 genera and 6 species were collected in Yinchuan and Wuzhong cities, Ningxia, among which Culex pipiens pallens was the dominant species with 3 050 individuals, accounting for 35.63%; Anopheles sinensis with 2 379 individuals, accounting for 27.79%; Culex tritaeniorhynchus with 1 489 individuals, accounting for 17.39%; Caspian Aedes with 1 468 individuals, accounting for 27.79%; Aedes vexans with 152 individuals, accounting for 1.78%; and Culex modestus with 23 individuals, accounting for 0.27%. JEV GIb type was detected in the specimens of Culex tritaeniorhynchus collected in Qingtongxia city. Conclusions:The dominant mosquito species in the surveyed areas of Ningxia are primarily Culex pipiens pallens, and JEV GIb virus was detected in Culex tritaeniorhynchus in Qingtongxia city. This study provides basic data for understanding the current status of mosquitoes and mosquito-borne viruses in the Ningxia region and offers scientific guidance for further public health prevention and control measures.

Objective:To explore the effect of integrating open laboratories into Medical Microbiology experimental teaching for students majoring in Medical Laboratory Technology to enhance teaching quality.Methods:A total of 58 students from the grade 2020 majoring in Medical Laboratory Technology at a medical college were selected as the control group,receiving traditional teaching methods without open laboratory contents.A total of 58 students from the grade 2021 majoring in Medical Laboratory Technology were selected as the experimental group,integrating open laboratory contents into Medical Microbiology experimental teaching.The impact of open laboratories on teaching quality in Medical Microbiology experimental teaching was evaluated through analysis of final examination scores and questionnaire surveys.Results:Compared with the control group,the experimental group showed significant improvements in experimental report scores,experimental theory assessment scores,practical skill assessment scores,classroom performance scores,and overall scores(P＜0.01).The learning interest,multi-channel learning,mastery of key concepts,and ability to apply and expand knowledge of the experimental group also significantly improved(P＜0.01).The satisfaction rate of the experimental group in terms of participation level,acquired knowledge,mastered skills,and improved learning outcomes was all above 90%.Conclusion:Integrating open laboratory content into Medical Microbiology experimental teaching for students majoring in Medical Laboratory Technology can effectively stimulate students'engagement,enhance teaching quality,and help students better understand and master professional knowledge.