AIM:To investigate the role of NFATc1 in vascular generation in the nude mice transplanted with human ovarian cancer SKOV3 cells.METHODS: NFATc1 expression was silenced by siRNA in SKOV3 cells.Human ovarian cancer transplantation nude mouse model was established by transplanting with SKOV3 cells in which the NFATc1 gene was silenced by siRNA technique.The expression of NFATc1, CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels was determined by RT-PCR, Western blotting and immunohistochemical staining.The tumor growth, angiogenesis and lymphangiogenesis were also observed.RESULTS:Over-expression of NFATc1 was observed in human ovarian cancer tissues.The silencing of NFATc1 expression by siRNA decreased tumorigenesis of transplanted ovarian cancer cells in the nude mice, reduced tumor vascular generation and inhibited the expression of CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels.CONCLUSION:NFATc1 is overexpressed in ovarian cancer.NFATc1 silencing regulates the tumor vascu-lar generation.NFATc1 thus has potential as a therapeutic target and for use in the diagnosis and evaluating prognosis of epithelial ovarian cancer.

OBJECTIVETo develop a rapid, simple, cost-effective, accurate and sensitive method for quantitative detection of mitochondrial DNA (mtDNA) 3243A→G mutation in order to provide reference for selecting the best detection method under different conditions.

METHODSGenomic DNA was extracted from peripheral leucocytes of 17 individuals from a Wenzhou family featuring maternally inherited diabetes and deafness (MIDD). Heteroplasmic level of mtDNA 3243A→G mutation was determined respectively with polymerase chain reaction-restriction fragment length polymorphism (PCR-RLFP), real time-amplification refractory mutation system-quantitative PCR (RT-ARMS-qPCR) and pyrosquencing. Eleven plasmids with various heteroplasmic levels of the 3243A→G mutation (ranging from 0 to 100%)were constructed as the standards. The reliability of above methods was compared by correlation coefficient based on observed and expected values.

RESULTSFor all three methods, measurement of the standards showed a linear correlation between the expected and detected values, i.e., PCR-RFLP (R(2)=0.828), RT-ARMS-qPCR (R(2)=0.998) and pyrosquencing (R(2)=0.997). For the MIDD family, it was consistent that there are 13 members carrying the A3243G mutation with different heteroplasmic levels. And there was no significant difference between the results by RT-ARMS-qPCR and pyrosquencing.

CONCLUSIONPCR-RFLP is not appropriate for the quantitative detection but could be used for early clinical screening. Both RT-ARMS-qPCR and pyrosquencing are suitable for the detection of low heteroplasmic level of A3243G mutation. Compared with pyrosquencing, RT-ARMS-qPCR is rapid, reliable and cost-effective, and is the best choice for detecting low mutation loads.

Adolescent ; Adult ; Base Sequence ; Child ; DNA, Mitochondrial ; genetics ; Deafness ; diagnosis ; genetics ; Diabetes Mellitus ; diagnosis ; genetics ; Female ; Genetic Testing ; methods ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Young Adult

Objective:To summarize the clinical characteristics, causes of misdiagnosis and preventive measures of infectious mononucleosis (IM) in children, and to improve the ability of clinicians in early diagnosis of IM in children.Methods:The clinical data of 468 children with IM in Xi′an Children′s Hospital from January 2018 to December 2021 were retrospectively analyzed, including general situation, disease onset, diagnosis and misdiagnosis.Results:Among the 468 children, 33 cases were clinically diagnosed and 435 cases were a definitely diagnosed; 281 males (60.04%) and 187 females (39.96%); the incidence rate was highest in preschool children (43.80%, 205/468) and in autumn (33.12%, 155/468). The first symptoms were fever (52.99%, 248/468), eyelid edema (15.38%,72/468) and neck mass (14.96%, 70/468). The fever rate was 90.38% (423/468), and the median time of first fever appearance was on the first (first, second) day of disease course, and the median duration of fever was 6 (4, 8) d. The median time of first visit was on the third (first, fifth) day of disease course, and the time of diagnosis was on the seventh (fifth, ninth) day of disease course. Blood routine examination showed that the proportion of white blood cell count increased was 51.92% (243/468), the proportion of lymphocytes increased was 61.75% (289/468), and the proportion of abnormal lymphocytes increased (≥10%) in peripheral blood was 58.97% (276/468). The lymphocyte subsets of 364 children were detected, the rate of helper T lymphocytes (Th cells) decreased was 80.22% (292/364), the rate of suppressor T lymphocytes (Ts cells) increased was 99.45% (362/364), the value and decreased rate of Th cells/Ts cells were 0.24 (0.16, 0.40) and 100.00% (364/364), rate of B lymphocytes decreased was 93.96% (342/364), rates of natural killer cells decreased and increased were 35.16% (128/364) and 0.55% (2/364). The misdiagnosis rate was 55.13% (258/468), and the misdiagnosis time was on the fifth (fourth, seventh) day of disease course. Among the 258 misdiagnosed children, 105 cases (40.70%) were misdiagnosed as upper respiratory tract infection, 65 cases (25.19%) as acute suppurative tonsillitis, 27 cases (10.47%) as acute cervical lymphadenitis or neck mass.Conclusions:Due to the complex and diverse clinical manifestations of IM in children, it is easy to be misdiagnosed in the early stage of the disease. So, it is necessary for clinicians to master the clinical characteristics of IM in children, constantly improve the level of diagnosis and treatment, and reduce the misdiagnosis rate.

Objective:To investigate the reliability and repeatability of 3 classification systems for coronal fractures of distal humerus.Methods:Included were the imaging data (X-ray and CT scan) of the 64 patients with distal humeral coronal fracture who had been admitted to the outpatient and emergency departments of Beijing Jishuitan Hospital from January, 2015 to December, 2021. Four orthopedic surgeons (1 chief surgeon, 1 associate chief surgeon, 1 attending surgeon, and 1 resident surgeon) served as observers who classified the fractures by their imaging data respectively according to the Bryan-Morrey, Ring and Dubberley classifications. Four weeks later when the imaging data of 64 patients were renumbered, the 4 orthopedic surgeons performed the same classifications again. Inter-observer and intra-observer Kappa values were calculated for the 3 classifications. Landis and Koch Kappa's Benchmark Scale was used to interpret the results.Results:Interobserver confidence: The average agreement of the Bryan-Morrey classification was 75.0%, and its Kappa value was 0.546, indicating moderate agreement. The average consistency of Ring classification was 74.7%, and its Kappa value was 0.702, indicating high consistency. The average consistency of Dubberley classification was 84.1%, and its Kappa value was 0.785, indicating high consistency. Intra-observer repeatability: The average consistency of Bryan-Morrey classification was 89.9% and its Kappa value was 0.824 for the same observer. The average consistency of Ring classification was 91.0%, and its Kappa value was 0.884. The average consistency of Dubberley classification was 91.8%, and its Kappa value was 0.888.Conclusions:The 3 classification systems for distal humeral coronal fractures all demonstrate high reproducibility. As the Dubberley classification system is relatively more reliable, it has a certain guiding value for the treatment of coronal fractures of the distal humerus. However, in clinical practice, an optimal surgical strategy should take into account various factors that may affect fracture typing.

Objective To analyze TCM master Yan Zhenghua's medication rules of ascending,descending,floating and sinking properties for the treatment of respiratory diseases,inherit his valuable academic experience.Methods Using four books edited by Professor Yan Zhenghua's disciples as the main source,Professor Yan Zhenghua's prescriptions for clinical treatment of respiratory diseases were systematically collected and analyzed.Statistical analysis was conducted on the patients'gender and age,differentiation of diseases and syndrome types of the prescriptions,as well as the properties of ascending,descending,floating and sinking,dosage,and commonly used pairs of Chinese materia medica.Results Totally 208 prescriptions were included in this study,involving 178 kinds of Chinese materia medica and 64 kinds of monarch drug,most of them were descending and sinking drug,and the whole prescription was mainly descending and sinking.On average,each prescription used 13.2 kinds of Chinese materia medica,and most dosage points were within the range of conventional dosage.Among later adding medicines,Houttuyniae Herba had the highest frequency of use.The medicinal pairs of Asteris Radix et Rhizoma-Cynanchi Stauntonii Rhizoma et Radix,Cynanchi Stauntonii Rhizoma et Radix-Stemonae Radix and Asteris Radix et Rhizoma-Stemonae Radix were often used.Armeniacae Semen Amarum,Fritillariae Thunbergii Bulbus,and Asteris Radix et Rhizoma were commonly used in those descending and sinking prescriptions.Conclusion In the treatment of respiratory diseases,Professor Yan Zhenghua prefers to use descending and sinking drugs with ascending and floating drugs and dual trend drugs to regulate qi activity,and has the characteristics of Menghe medical school,which is"mild and flexible medication".

Objective:To design a novel electromagnetic ejection device for endoscopic suturing to achieve continuous deployment of suture nails.Methods:An electromagnetic ejection device and its accompanying suture nail structure were designed and a prototype was fabricated based on electromagnetic ejection principles. A finite element model of the electromagnetic ejection device was constructed to study the effects of armature-coil center distance and different driving voltages on suture nail ejection speed. An experimental platform for testing electromagnetic ejection velocity was constructed, and a high-speed camera was used to detect the ejection velocity. A platform for the suture embedding experiment was built to measure the effects of different voltages on the inserting speed of suture into the gastric wall tissue. A platform for a suture extraction force experiment was built to evaluate the extraction force of sutures embedded in tissues under different driving voltages.Results:A suture nail structure and electromagnetic ejection device were designed, and a prototype was fabricated. The ejection velocity increased and then decreased with the increase of the armature-coil center distance, and the maximum ejection velocity was 15.81 m/s at the center distance of 18 mm. At this distance, the voltage was linearly related to the ejection velocity, and the experimental values of the staple basically coincided with the simulated values. When the driving voltage was in the range of 150 to 180 V, the suture nails could successfully insert in the tissues, and the 180 V voltage group had a greater insertion depth. The extraction force of the suture nails at 120, 150, 180, and 210 V voltages were (0.49 ± 0.19), (1.14 ± 0.19), (1.23 ± 0.15), and (1.85 ± 0.31) N, respectively.Conclusions:A novel electromagnetic ejection device for endoscopic suturing is proposed that is capable of continuous firing of suture nails. This device provides a new long-distance driving method for intelligent, minimally invasive surgical instruments.