ObjectiveTo investigate the application of endoscopy in obtaining the great saphenous vein （GSV） during coronary artery bypass grafting （CABG） and explore the learning curve， with a particular focus on common challenges encountered during the learning process and their impact on early clinical outcomes. MethodsA retrospective analysis was conducted on clinical data from 83 patients who underwent off-pump CABG with endoscopic GSV harvesting at the First Affiliated Hospital of Zhengzhou University from July 2013 to April 2014. Patients were categorized into four groups based on the chronological order of their hospitalization： Group A （novice group， n=20）， Group B （proficient group， n=20）， Group C （progressive group， n=20）， and Group D （mature group， n=23）. Differences in perioperative and midterm follow-up outcomes among the groups were analyzed to determine the learning curve period. ResultsThe study population had a mean age of （60.22±8.06） years and a mean body weight of （69.77±11.66） kg. Comorbidities included hypertension （24 cases）， diabetes （26 cases）， and subacute cerebral infarction （14 cases）. The novice group exhibited significantly shorter GSV length-to-harvest time ratio relative to the other three groups （P<0.001） and a significantly higher incidence of main vein damage （P=0.006）. However， there was no statistically significant difference in graft patency at the 1-year follow-up. ConclusionThorough and reliable technical training in endoscopic GSV harvesting is essential to minimize vascular injury caused by novice operators. Approximately 20 cases of hands-on experience and a careful self-analysis of procedural challenges are likely required to achieve proficiency in GSV harvesting.

Objective:To investigate the clinical efficacy of ultrasound-guided vacuum-assisted excision (VAE) in the treatment of breast benign phyllode tumor and the risk factors of local recurrence after operation.Methods:A total of 256 cases of patients with breast benign phyllode tumor admitted to Yuncheng Central Hospital from January 2012 to January 2020 were included in the retrospective study, they were all female, of which 160 cases received ultrasound-guided VAE surgical treatment and 96 cases received open surgical treatment. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test was used for comparison between groups. The Chi-square test or Fisher exact probability method was used to compare the data groups. Univariate analysis was performed on the factors related to the local recurrence of benign phyllode tumor after surgery, and then the factors with statistically significant differences in the univariate analysis were further incorporated into multivariate Logistic regression analysis. Results:Univariate analysis results showed that the maximum diameter of tumor ≥25 mm and the history of ipsilateral breast fibroadenoma were associated with postoperative local recurrence of breast benign phyllodes tumor ( P<0.05). Multivariate Logistic regression analysis results showed that tumor diameter ≥25 mm was an independent risk factor for postoperative local recurrence of breast benign phyllodes tumor ( P<0.05). Conclusion:Ultrasound-guided VAE in the treatment of breast benign phyllodes tumor patients with maximum diameter less than 25 mm can reduce the postoperative local recurrence rate, and patients with tumor maximum diameter greater than 25 mm should have the higher local recurrence risk.

Objective:To screen and diagnose cervical lesions in middle-aged and elderly postmenopausal women and analyze the characteristics of cervical lesions in middle-aged and elderly women.Methods:A retrospective analysis was conducted on the screening data of 1 337 postmenopausal women aged between 50 and 86 years who visited the outpatient clinics of the gynecology department of Beijing Hospital between January, 2015 and December, 2019.Participants were divided into groups based on age, with 787 in the 50-59 years old group, 483 in the 60-69 years old group, 60 in the 70-79 years old group, and 7 in the 80-89 years old group.Exfoliated cervical cells were collected, and high-risk human papillomavirus(HPV)detection and liquid-based cervical cytology examination were performed to confirm the diagnosis based on the pathological results of cervical biopsy.Cervical lesions were analyzed.Results:53.0%(708)of the 1 337 enrolled patients made their hospital visits because of symptoms.There were 666 patients(49.8%)with cervical lesions, including 357 with low-grade squamous intraepithelial lesions(LSIL), 252 with high-grade squamous intraepithelial lesions(HSIL)and 57(4.3%)with cervical cancer.The 70-79 group showed the highest incidence of HSIL at 30.0%(18/60), while the 50~59 group had the highest incidence of LSIL at 30.6%(241/787), and there were significant differences in different types of cervical lesions between different age groups( χ2=28.919, P<0.01). Liquid-based cervical cytology examination revealed a positive rate of 51.6%(166/322)for LSIL, a positive rate of 70.8%(143/202)for HSIL, and a rate of 71.4%(25/35)for cervical cancer.There were significant differences in LSIL, HSIL and other above-mentioned positive rates of lesions( P<0.05). In the cervical malignancy group, cytological high-grade squamous epithelial lesions made up 51.4%(18/35). Intraepithelial lesions or cancerous lesions represented 29.2%(59/202)in the HSIL group.The rate of intraepithelial lesions or cancerous lesions in the LSIL group was 48.4%(156/322), with statistically significance in these differences( χ2=118.806, P<0.01). Chronic cervicitis was found in 52.9%(109)of 206 patients with atypical squamous cells and positive HPV, and in 76.2%(16)of 21 patients without HPV.HPV examination was completed in 1 301 patients, with 69.4%(903)having infections by one HPV strain and 30.6%(398)having infections by mixed HPV strains.Biopsy results showed that the rates of HPV infections in LSIL, HSIL and cancer patients were 84.6%(302/357), 95.9(185/193)and 100%(54/54), respectively, and the differences in these pathological results were statistically significant( P<0.01). HPV16 was the most common subtype in single HPV strain infections.The most common subtypes of HPV infections in cervical lesions with HSIL and above were HPV16 and HPV18.The agreement rate was 52.4%(187/357)for LSIL, 60.7%(153/252)for HSIL, and 64.9%(37/57)for cervical cancer between diagnoses based on cervical biopsy and those based on colposcopy.As the severity of lesions increased, the accuracy of colposcopy also increased.Differences between colposcopic diagnosis and pathological diagnosis on different types of cervical lesions were statistically significant( χ2=550.382, P<0.01).1 310 patients underwent endocervical curettage, identifying 120 cases of HSIL and 39 cases of cervical cancer, with a detection rate of 55.8%(159/285). Conclusions:The incidence of cervical cancer is high in elderly women.The incidences of high-grade lesions and cervical cancer in patients with atypical squamous cells plus HPV infections in the cervix are also high.Postmenopausal patients with cervical lesions are mainly infected by a single HPV subtype, with subtype 16 being the most common one.With increasing severity of lesions, the accuracy of colposcopy-based diagnosis increases and the rate of missed diagnosis decreases.Cervical curettage is an important means to reduce missed diagnosis with colposcopy.

Hypoxic microenvironment is a common phenomenon of liver diseases and runs through the whole process of the development and progression of liver diseases. In recent years, the research on liver fibrosis and hypoxic microenvironment of hepatocellular carcinoma has attracted more and more attention. Hypoxia-inducible factor (HIF) is the most important hypoxic stress factor found at present. This article reviews the characteristics of hypoxic microenvironment in the liver and liver diseases and further elaborates on the association of HIF overexpression with hepatocyte damage, formation of fibrosis, and malignant transformation of hepatocytes.

Objective Studying the differential expression of exosomal miRNAs between androgen-dependent prostate cancer cell and androgen-independent prostate cancer cell,in order to further elucidate the mechanism of androgen-independent prostate cancer and find new targets for its treatment.Methods Prostate cancer androgen-dependent LNCaP cell and prostate cancer androgen-independent LNCaP-AI + F cell (induced by androgen and flutamide) were selected as the study subjects.Illumina HiSeqTM 2500 was used to perform high-throughput sequencing between the two groups.The differentially expressed exosomal miRNAs was verified by quantitative real-time PCR(qRT-PCR).The difference was statistically significant by t-test.Results Through the analysis of high-throughput sequencing results,thirteen molecules were screened increased in extracellular exosomes of the androgen-independent cell,including miR-7-5p,let-7a-5p,miR-375,miR-423-3p,miR-378a-3p,and miR-92a-3p,etc.Among them,miR-7-5p was verified by quantitative real-time PCR to be up-regulated by 19.52-fold (t=9.857,P=0.001).Conclusion Differentially expressed exosomal miRNAs may predict the development of androgen-independent prostate cancer and may further regulate the development of androgen-independent prostate cancer.

Objective: To investigate the effects of prostate cancer exosomes on the migration and invasion ability of stromal cells (WPMY-1), and to explore its mechanism. Methods: Exosomes in LNCaP-AI+F prostate cancer cell supernatant were isolated by ultracentrifugation and the typical structure of exosome was captured by electron microscope. The particle size distribution was analyzed by Zetaview, and Wb was used to identify the marker proteins and other proteins.After co-incubation of WPMY-1 cells and prostate cancer exosomes (40 µg/ml), laser confocal microscope was used to observe the uptake of PKH67-labeled exosomes by WPMY-1 cells; Transwell assay was used to detect the migration and invasion ability of WPMY-1 cells; qPCR was performed to detect the expression of three cancer-associated fibroblast (CAF)-related molecules (IL-8, PDGFB and MMP9) at mRNA level; and the phosphorylation of EGFR and ERK1/2 was analyzed by Wb. Results: Typical cup-shaped structure of exosomes was observed under electron microscope. The Zetaview results showed that the particle size distribution was concentrated at about 100 nm. The expression of exosome marker proteins CD63 andALIX further verified that the isolated particles were exosomes. Besides, EGFR, HER2 and SRC, which were related to the progression of prostate cancer, were also enriched in exosomes. After co-incubation, confocal microscope imaging showed a number of PKH67 labeled exosomes in recipient WPMY-1 cells. Transwell experiments showed that exosomes could significantly enhance the migration and invasion ability of WPMY-1 cells (all P<0.01). Compared with the control group, increased secretion of IL-8, PDGFB and MMP9 was observed after exosome treatment (40 µg/ml) (P<0.05 or P<0.01). Wb indicated that exosomes could promote the phosphorylation of EGFR and ERK1/2 of WPMY-1 cells (P<0.01). Conclusion: Prostate cancer cell exosomes could act on the stromal cell WPMY-1 to highly express multiple CAF-related molecules, promote the phosphorylation of EGFR and ERK1/2 and enhance the migration and invasion ability of WPMY-1 cells.

Objective: To optimize the existing methods of isolation and purification for exosomes from urine and explore the effects of different storage conditions on the content of exosomal RNA in urine. Methods: The exosomes in human urine samples were extracted by different precipitation method, i.e., precipitation following first concentrating and direct precipitation, respectively, and the separation efficiency and cost of the two methods were compared. ExoQuick-TCTM precipitation kit was used to extract exosomes. Nanoparticle tracking analysis technique (NTA) was used to detect the concentration and particle size distribution of exosome. Dynamic light scattering (DLS) was used to detect the potential of exosome. Transmission electron microscopy (TEM) was used to observe morphology of exosomes. western blot was used to analyze the exosomal marker molecules CD63 and Alix. The extraction method of the precipitation following first concentrating was used to verify the reliability of the optimized method in 10 clinical urine samples . Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of exosomal RNA marker let-7c and PSA mRNA in the urinary exosomes from 20 patients with prostate cancer after repeated freeze-thaw (0 [i.e., fresh], 1 , 3 and 5 times) and 9 patients with prostate cancer frozen at -80 ℃ for different time (0 [i.e., fresh], 1, 2 and 4 weeks), and were statistically analyzed by Wilcoxon rank sum test for differences between the 2 groups. Results: The size distribution of exosomes extracted by the two methods was 30 to 150 nm by NTA, both of which were displayed as single peaks. The results of DLS showed that the potentials of exosome extracted by the two methods were negative values. The size of the exosomes extracted by the two methods was consistent observed under TEM namely the diameter distribution was 30 to 150 nm. western blot analysis confirmed that CD63 and Alix, the exosome labeling molecules, existed in the optimized method. The concentration of exosomes extracted from the 10 urine samples all reached 10 9 to 10 11 particles/mL. The contents of let-7c and PSA mRNA in exosomes decreased significantly after 5 freeze-thaw cycles, and the Z values were -1.79 and -1.73, respectively (P＜0.05). The RNA content of the exosomes remained stable after freezing at -80 ℃ for 1 month. Conclusion The optimized exosome extraction method could reduce greatly the cost under the premises of ensuring the concentration and quality of exosomes. The isolated exosomes may keep stable RNA content after freezing at -80 ℃ for a short time, but could not be frozen and thawed repeatedly for more than 5 times.

The design of research protocol and quality control are the key to ensure the quality of clinical trial. A randomized clinical trial regarding the effects of medication combined with acupuncture on live birth rate in polycystic ovary syndrome (PCOS), which was initially designed as a comparative effectiveness research, then added with an acupuncture control group and finally became a factorial analysis, is taken as an example to explain the protocol design and optimization process, demonstrating the high level of methodology design and international recognization. By a series of measurements, such as unified purchase of acupuncture equipment, multiple trainings and assessments for acupuncturists' knowledge and operation standardization, in-site supervision of local center experts, the standard operation of acupuncture could be ensured and the credibility and scientificity of research results could be improved.

Objective To investigate the molecular epidemiological characteristics and homology of Staphylococcus aureus (S. aureus)isolated from patients in intensive care units (ICUs)of a hospital,so as to provide laboratory basis for the effective control of healthcare-associated infection(HAI). Methods 62 S. aureus strains isolated from various specimens from ICU patients with infection in March-August 2013 were collected,7 housekeeping genes were amplified with polymerase chain reaction (PCR),the amplified products were sequenced,ST typing of strains was performed by multilocus sequence typing (MLST ), phylogenetic analysis of ST typing was conducted. Results 62S. aureus strains were amplified specific product of 7 housekeeping genes;there were 10 ST genotypes, in which 2 ST genotypes(STn1and STn2)were first discovered,1 ST genotype(ST675)was first discovered domes-tically. ST239 was the main ST type of S. aureus from ICU patients in this hospital,accounting for 74.20% ,which distributed in 6 ICUs,ST5 distributed in 3 ICUs. 62 strains formed 7 main branches in the phylogenetic tree,55 (88.71% )MRSA strains were detected. Conclusion S. aureus isolated from hospital ICUs has some homology, and the small number of types showed the trend of concentrated distribution.

Neural prosthesis control system is based on brain-computer interface and functional electrical stimulation technology, by an-alyzing the electroencephalograph control commands directly into the muscle system or an external device, which compensated efferent pathway from the brain-spinal cord, and recovered motor function of patients with cervical spinal cord injury. This paper described the basic structure, working principle and key technology of neural prosthetic system, summarized the application, problems and prospects of neural prosthetic technology in the rehabilitation of cervical spinal cord injury.