Objective We aimed to explore the mechanism by which Bushen Yangjing Soup improves elderly infertility based on oocyte mitochondrion.Methods C57BL/6J mice were used,including 15 young females(6-8 weeks old),45 elderly females(10 months old),and ten young males(6-8 weeks old).The female 6-8-week-old young mice were assigned to the young group,and the female 10-month-old elderly mice were randomly divided into the model group,the dehydroepiandrosterone(DHEA)group,and the Bushen Yangjing Soup group.Mice in the DHEA and Bushen Yangjing Soup groups were given DHEA[11.26 mg/(kg·d)]and Bushen Yangjing Soup[11.41 g/(kg·d)].Mice in the other groups were given normal saline.After 45 days,oocytes were collected,the number of oocytes and the proportion of mature oocytes were recorded,and in vitro fertilization(IVF)was carried out,followed by calculation of the successful fertilization rate.Real-time PCR was used to determine the copy number of mitochondrial DNA in oocytes,mito-tracker green was used to observe the distribution of mitochondria,and the mitochondrial membrane potential(MMP)of oocytes was determined using the JC-1 kit.The mRNA and protein levels of mitofusin 2(Mfn2)in oocytes were determined by real-time PCR and Western blotting,respectively.Results Compared with the young group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly reduced(P＜0.01),the copy number of mitochondrial DNA in oocytes was decreased,the rate of abnormal mitochondrial distribution was increased,the MMP was decreased(P＜0.01),and the mRNA and protein levels of Mfn2 were decreased(P＜0.01)in the elderly model mice.Compared with the elderly model group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly increased(P＜0.05),the copy number of mitochondrial DNA in oocytes was increased(P＜0.05),the rate of abnormal mitochondrial distribution was decreased,the MMP was increased(P＜0.05),and the mRNA and protein levels of Mfn2 were increased(P＜0.01)in the Bushen Yangjing Soup group.Compared with the DHEA group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were increased,the copy number of mitochondrial DNA in oocytes was increased,the rate of abnormal mitochondrial distribution was decreased,the MMP was increased,the mRNA and protein levels of Mfn2 were increased in the Bushen Yangjing Soup group,and there was a significant difference in the number of oocytes and the expression of Mfn2 protein(P＜0.05).Conclusion Bushen Yangjing Soup may improve the quantity,distribution,and function of oocyte mitochondria by increasing the mRNA and protein levels of Mfn2,thereby improving oocyte quality.This is one of the possible mechanisms for clinical application of Bushen Yangjing Soup to improve ovarian function and treat elderly infertility.

Objective:To understand molecular characteristics and antibiotic resistance of Clostridioides ( C.) difficile isolated from children in China, and provide data support the development of disease risk assessment and burden studies. Methods:A total of 155 strains of C. difficile isolated from children aged <12 years in 14 provinces (autonomous regions, municipalities) in China from 2010 to 2023 were used for the analyses on molecular characteristics and antibiotic resistance of C. difficile by PCR and drug susceptibility test. Results:A total of 26 sequence types (STs) and 18 ribotypes (RTs) were identified in the 155 C. difficile isolates, in which ST3 (20.65%), ST54 (16.13%), ST35 (12.90%), and RT012/ICDC007 (14.84%), RT001/ICDC001 (11.61%), RT046/ICDC018 (8.39%) were the most common. One highly virulent strain with RT078 and 27 non-toxin-producing strains were also found; the predominant toxin gene was tcdA+ tcdB+ cdt-. All the strains were sensitive to metronidazole and vancomycin, and there were 29 multidrug-resistant strains, in which 1 strain was resistant to all the seven antibiotics except for vancomycin and metronidazole. Conclusions:Molecular characteristics and antibiotic resistance of C. difficile in children were similar to those in whole population in China, but there were regional distribution differences. It is necessary to strengthen the routine drug-resistance surveillance for C. difficile infection in children in China.

Objective:To analyze the accuracy of lung ultrasound and chest X-ray in the diagnosis of neonatal pulmonary disease.Methods:We prospectively collected newborns that needed chest X-ray examination to diagnose pulmonary disease from twelve neonatal intensive care units across the country between June 2019 and April 2020.Each newborn was examined by lung ultrasound within two hours after chest X-ray examination.All chest X-ray and lung ultrasound images were independently read by a radiologist and a sonographer.When there was a disagreement, a panel of two experienced physicians made a final diagnosis based on the clinical history, chest X-ray and lung ultrasound images.Results:A total of 1 100 newborns were enrolled in our study.The diagnostic agreement between chest X-ray and lung ultrasound(Cohen′s kappa coefficient=0.347) was fair.Lung ultrasound(area under the curve=0.778; 95% CI 0.753-0.803) performed significantly better than chest X-ray(area under the curve=0.513; 95% CI 0.483-0.543) in the diagnosis of transient tachypnea of the newborn( P<0.001). The accuracy of lung ultrasound in diagnosing neonatal respiratory distress syndrome, meconium aspiration syndrome, pneumonia and neonatal pulmonary atelectasis was similar to that of chest X-ray. Conclusion:Lung ultrasound, as a low-cost, simple and radiation-free auxiliary examination method, has a diagnostic accuracy close to or even better than that of chest X-ray, which may replace chest X-ray in the diagnosis of some neonatal lung diseases.It should be noted that both chest X-ray and lung ultrasound can only be used as auxiliary means for the diagnosis of lung diseases, and it is necessary to combine imaging with the clinical history and presentation.

Objective To design an automated detection protocol for morphological detection of sperm head based on deep learning and evaluate its efficiency,accuracy and reliability.Methods Fourteen pictures for each of 1 000 samples were analyzed by using the pre-trained YOLO target detection model and VGG16 classification model.At least 200 sperm were detected for each sample.Equal amount of samples were analyzed by manual microscope examination,and the efficiency,accuracy and correlation between the two methods were compared.Results The morphology of sperm heads which were manually classified but untrained was detected by pre-trained classification model,and the prediction accuracy reached to 95.5％.The detection time for clinical each sample was only 10 seconds,and its accuracy and efficiency were higher than those of manual microscope examination.The percentages of the sperms with normal morphology were significantly positively correlated(r=0.84,P＜0.01)in the detections of both the methods.Conclusion The detec-tion protocol proposed in this study can greatly improve work efficiency,and its reliability and accuracy exceed those of manual micro-scope examination.

Objective:To investigate the effects of CT images reconstructed using different field of view (FOV) sizes on the automatic segmentation of organs at risk and dose calculation accuracy in radiotherapy after radical mastectomy.Methods:Under the same scanning conditions, CT values-electron density conversion curves were established by reconstructing the original CT images of a phantom placed at the isocenter and extended FOV (eFOV) positions using FOV sizes of 50, 60, 70 and 80 cm. Then, these curves were compared. A standard phantom with a known volume was scanned, and the automatic segmentation result of the phantom on CT images reconstructed using different FOV sizes was compared. A total of 30 patients in Guangdong Second Provincial General Hospital from January 2020 to June 2022 with breast cancer were randomly selected. Through simulated positioning, their CT images were reconstructed using different FOV sizes for the purpose of automatic segmentation of organs at risk, followed by comparison between the outcomes of automatic segmentation and physicians′segmentation. The treatment plan established based on CT images reconstructed using a FOV size of 50 cm (FOV 50 images for short) was applied to CT images reconstructed using FOV sizes of 60, 70 and 80 cm (FOV 60, FOV 70 and FOV 80 images for short) for dose calculation, and the dose calculation result were compared. Results:The CT values - electron density conversion curves derived from CT images reconstructed using different FOV sizes were roughly consistent. At the isocenter, the difference between the segmented volume and actual volume of the standard phantom increased up to a maximum of 6 cm 3 (4.8%) with an increase in the FOV size. As indicated by the automatic segmentation result, the segmentation accuracy of the spinal cord, trachea, esophagus, thyroid, healthy mammary gland, and skin decreased with an increase in the FOV size ( t = -28.43-8.23, P < 0.05). The comparison of dose calculated based on CT images reconstructed using different FOV sizes showed that there was no statistically significant differences( P>0.05) in the dose to target volume ( V95) and the maximum and average doses in the supraclavicular lymph node region, as well as the dose to organs at risk. The coverage for planned target volume decreased with an increase in the FOV size, with a maximum difference of 4.06%. Conclusions:It is recommended that, for radiotherapy after radical mastectomy, FOV 50 images should be selected for the automatic segmentation of organs at risk, CT-values-electron density conversion curves should be established based on the electron density phantom images of the eFOV region, and the eFOV 80 images should be preferred for dose calculation.

Clostridioides difficile infection (CDI) is an infectious disease with fever, abdominal pain and diarrhea as the main clinical manifestations. At present, CDI is mainly treated with antibiotics and faecal microbiota transplantation. As recurrent and refractory CDI continues to increase, it is important to seek a more effective alternative therapy. However, many of the studies on the prevention and control of CDI by probiotics are still in the early stage. This paper summarized the research on the types, mechanisms and technical means of probiotics in the treatment of CDI.

Clostridioides difficile is a key pathogen of antibiotic related diarrhea and hospital associated infection, causing several outbreaks in Europe and North Americans and resulting in severe disease burden. However, the standardized diagnostic principle and detection specifications in C. difficile infection (CDI) survey are limited in China, and the infection rate and disease burden of CDI in China are unclear. Therefore, National Institute for Communicable Disease Control and Prevention,National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, together with another 11 institutions, draft the group standard entitled "Diagnosis of Clostridium difficile infection (T/CPMA 008-2020)" of Chinese Preventive Medicine Association. Based on the principle of "legality, scientificity, advancement, and feasibility", this standard clarifies risk factors, diagnosis principles, diagnoses and differential diagnoses in order to improve the accuracy of CDI diagnosis in clinical practice, guide the surveillance for CDI, and understand the infection rate and disease burden of CDI in China .

Objective:To study clinical pathological characteristics, immunohistochemical, molecular genetical changes and prognosis in pediatric eosinophilic solid and cystic renal cell carcinoma (ESC RCC) with TSC2 gene mutations.Methods:The tissue samples were collected from two pediatric ESC RCC patients between 2017 and 2018. The tissues were subjected to histological examination and immunohistochemistry using EnVision system. The TFE3, TFEB gene rearrangements were tested using FISH and molecular genetic study. The paraffin sections were used for DNA extraction, PCR amplification and NGS sequencing.Results:The two patients with ESC RCC were both male, aged at 9 years and 8 months, and 13 years, respectively. The tumors were from the right kidney, 5 cm and 7 cm in size, respectively, with solid and cystic changes in cross section, and grey-reddish or grey-whitish fish meat appearance. Microscopic observation revealed the tumors had fibrous capsules, which were infiltrated by the tumor cells. The tumor cells were diffusely distributed, round-shaped, or polygon-shaped, and had voluminous cytoplasm, eosinophilic cytoplasm, various sizes of vacuoles and clear cell-like appearance. There were papillary structures in some areas, with visible fiber septa. The nuclei were round and vesicular, with multi-nucleated cells and megakaryocytes. The mitoses were not seen. A few cystic structures were visible in different sizes, and capsule walls were covered with a single layer of spike-like tumor cells. Thick-walled blood vessels were seen in the stroma, with focal lymphocytic infiltration, eosinophilic necrosis, calcifications and cholesterol crystals. Immunohistochemistry of the tumor cells was positive for PAX8 (diffuse), CK20 (focal), CKpan (focal), CK10 (1 focal, 1 diffuse), INI1, vimentin, CD68, and Ki-67 (5%~10%); the tumor cells were negative for HMB45, S-100, Melan A, p53, desmin, TFE3, CK7, CK19, EMA, CD56, CgA, Syn, CD30, CD117, WT1 and SMA. Molecular genetic study showed that TFE3 and TFEB gene rearrangements were not detected by FISH. NGS sequencing showed TSC2 p.Lys574Ter (0.198) was found in patient one and TSC2 p.Arg406Ter (0.355) in patient two.Conclusions:ESC RCC in children is a rare disease, and can be misdiagnosed easily. It has unique pathological characteristics, and immunohistochemical, molecular and genetic changes. The prognosis is relatively good.

Objective: To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China. Methods: Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency. Results: Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance. Conclusions The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories.

Objective To summarize the clinical experience and efficacy of surgical treatment for Stanford type A aortic dissection leading to acute lower limb ischemia.Methods From January 2014 to January 2018,12 patients with severe lower limb ischemia caused by acute type A aortic dissection were treated with Suns surgery.Among them,11 patients were treated with restoration of lower limb blood supply preferentially,including 10 cases of femoral artery bypass and 1 case of abdominal aorta-iliac artery stent graft implantation.Another case was treated with ascending aorta-femoral artery bypass after Sung surgery.Results 3 cases died of ischemia and necrosis of the lower extremities.Two of them died of multiple organ failure due to amputation and one died of low cardiac output due to refractory acidosis.Acute renal failure performed bedside CRRT in 5 patients and ECMO in 1 patient.The remaining 9 patients were discharged from the hospital and the symptoms of lower limb ischemia disappeared.After an average follow-up of 23 months,the re-examination of the aorta CTA showed that the bypass artery was unobstructed and the distal femoral artery was well developed.One patient infecting vascular prosthesis was cured by taking out the unit.Conclusion For acute lower limb ischemia caused by type A aortic dissection,blood flow of lower extremities should be restored as soon as possible to reduce mortality and complications.Femoral artery bypass and abdominal aorta-iliac arterial repair are simple and effective in reconstructing lower limb blood supply.