In black tea,theaflavins (TFs) are one important class of substances that determine sensory quality and have significant medicinal activities. In addition to the four kinds of common TFs,there may be many other theaflavin analogues (TFAs) with similar chemical structures in tea,but the study on them is very limited. Based on the characteristic sub-structure,mass spectrometry (MS) and MS/MS information,a method for screening and annotation of TFAs from the complex ultra high performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) data was proposed in this work. By analyzing the oxidation and polymerization process of a few TFs,the theoretical reaction model of TFs were summarized,which was used to calculate the precursor ion values of potential TFAs. Meanwhile,the diagnostic fragmentation ions and neutral loss of TFAs according to the fragmentation pathways obtained from chemical standards or documented in literatures were summarized. As a result,36 kinds of compounds were successfully annotated based on the calculated precursor ion values and the MS fragmentation patterns,among which 6 kinds of compounds were reported for the first time in tea. In vitro synthesis experiments were carried out to verified the annotation results. Based on the results of quantitation of 36 kinds of TFAs,a partial least squares-discriminant analysis model was used to investigate the changes of these components during black tea manufacturing. The results indicated that these novel TFAs could be used to effectively distinguish the black tea samples before and after fermentation.

The current situation of childrearing in the elderly and low birth rate showed that the prevention and treatment of infertility risk caused by the decline of male reproductive function are increasingly becoming prominent.The decline of male reproductive function is closely related to the hypothalamic-pituitary-gonadal axis,oxygen free radicals,blood-testis barrier,and the apoptosis of spermatogenic cells.Traditional Chinese medicine believes that the kidney stores essence and is in charge of reproduction,and the decline of reproductive function is closely related to the imbalance of yin and yang of the kidney.Therefore,kidney-supplementing method is helpful for treating the decline of reproductive function caused by kidney deficiency.Kidney-supplementing prescriptions were effective on systematically regulating the function of the hypothalamic-pituitary-gonadal axis,thus to significantly delay the decline of reproductive function,were effective on removing excessive oxygen free radicals in the body and protecting cells from oxidative damage,and were effective on improving spermatogenesis by restoring the blood-testis barrier and regulating the apoptosis of spermatogenic cells.The representative kidney-supplementing prescriptions for improving the decline of reproductive function caused by kidney deficiency include Wuzi Yanzong Pills,Shenqi Pills,Yougui Pills,Liuwei Dihuang Pills,Zuogui Pills and Guilu Erxian Soft Extract.The above formulas presented no single therapeutic mechanism but multiple-way,multiple-level and multiple-target mechanism,and the formulas can comprehensively regulate the internal environment of the body,so as to achieve the purpose of improving male reproductive function.The kidney-supplementing therapy of traditional Chinese medicine plays an irreplaceable role in the treatment of male reproductive function decline.The in-depth study of its therapeutic mechanism will provide guide for clinical practice and will supply theoretical basis for improving male reproductive function decline.

ObjectivesTo investigate the ultrasound features and etiological distribution of non-immune hydrops fetalis. MethodsA total of 232 cases of diagnosed non-immune hydrops fetalis were recruited from December 2012 to January 2019 in The First Affiliated Hospital of Sun Yat-sen University. The ultrasound features and the results of prenatal diagnosis of hydrops fetalis were retrospectively analyzed. Results1. Non-immune hydrops fetalis was often associated with TTTS stage Ⅳ （50/232， 21.55%）； skin edema （159/232， 68.53%） was the mostly identified fluid collection； the most frequently combined malformations were anomalies of the cardiovascular system （15/232， 6.47%）. 2. Totally 185 cases accepted further prenatal genetic test and the abnormal detection rate was 40.54% （75/185）， while the abnormal detection rate of chromosome examination （including chromosome karyotype analysis and CMA） was 26.49% （49/185）. The abnormal detection rate of isolated NIHF was lower than that of non-isolated NIHF （32.64% vs 68.29%， p< 0.05）. Chromosome karyotype analysis was only performed in 57 cases， and abnormalities were detected in 14 cases （24.56%）. CMA only was performed in 31 cases and abnormalities were detected in 13 cases （41.94%）. Both Chromosome karyotype analysis and CMA were performed in 88 cases. Variation was detected in 22 cases （25%）， 3 cases （3.41%） showed abnormalities detected only by Chromosome karyotype analysis， while 6 cases （6.82%） had abnormalities detected only by CMA. The extra detection rate of CMA was 3.41% （3 cases） compared with Chromosome karyotype analysis. Variation was both detected by Chromosomal karyotype analysis and CMA in 13 cases （14.77%）. In this study， 30 cases of variation were detected by Chromosomal karyotype analysis （30/145， 20.69%）. The most common was 45， X （19/145， 13.10%）， followed by 47， XX， + 21 / 47， XY， + 21 （3/145， 2.07%）. CMA detected 32 cases of variation， including 12 cases of CNVs （10.08%）， 4 of which were pathogenic CNVs （3.36%）. Genetic analysis detected abnormalities in 26 of 27 cases. The most common abnormality in gene detection cases was αThalassemia --^SEA/--^SEA （21/26， 80.77%）， followed by PTPN11 gene mutation （2/26，7.69%）. 3. The causes of genetically normal NIHF included fetal-maternal blood transfusion， infection and unexplained anemia. ConclusionsThe most common etiology of non-immune hydrops fetalis is TTTS stage Ⅳ and the most frequently associated malformations are cardiovascular system abnormalities in our center. The most common abnormal karyotypes of non-immune hydrops fetalis are 45， X. CMA offers extra detection rate compared with Chromosome karyotype analysis in NIHF. The most common monogenic disease is αThalassemia--^SEA/--^SEA. The genetic abnormality detection rate of non-isolated NIHF is higher.

Objective: To investigate the daily variation of LPR and the significance of 48-hour oropharyngeal pH monitoring in the diagnosis of LPRD. Methods: 72 subjects with suspected LPRD who were treated in our department from June 2018 to June 2021 were included. All patients were hospitalized to complete continuous 48-hour oropharyngeal Dx-pH monitoring. The consistency of Ryan index and W index and the correlation of various reflux parameters between the first and second 24-hour were compared. SPSS 24.0 was used for statistical analysis. Results: All 72 subjects successfully completed 48-hour oropharyngeal Dx-pH monitoring. Ryan index was positive in 11 cases (15.2%) in the first 24-hour, in 17 cases (23.6%) in the second 24-hour, in 5 cases (6.9%) both first and second, and in 23 cases (31.9%) in either 24-hour, Kappa=0.211 (P=0.064), 18 cases (25%) had inconsistent results of the first 24-hour and the second 24-hour, and there was no significant difference in the positive rate between the first and second (P=0.234). The number of positive cases in 48-hour monitoring increased by 109.1% compared with 24-hour monitoring.For W index, 49 cases (68.1%) were positive in the first 24-hourf 53 cases (73.6%) were positive in the second 24-hour, 42 cases (58.3%) were positive both first and second, and 58 cases (80.6%) were positive in either 24-hour, Kappa=0.477 (P<0.001), 16 cases (22.2%) had inconsistent results of the first and second, and there was no significant difference in the positive rate between the first and second (P=0.804). The number of positive cases in 48-hour monitoring increased by 18.4% compared with 24-hour monitoring. There was no significant difference in all the reflux parameters of first and second (P>0.05). The correlation comparison showed that the correlation of various reflux parameters in the upright position was lower than that in the supine position. Conclusion: Laryngeal reflux has daily variability. Extending the monitoring time of Dx-pH to 48-hour can help reduce the missed diagnosis caused by daily variability; the use of W index can reduce the influence of daily variability on the diagnostic results of LPRD.
Humans ; Laryngopharyngeal Reflux/diagnosis* ; Hydrogen-Ion Concentration ; Hypopharynx ; Larynx

We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.
Exosomes ; Humans ; MicroRNAs/genetics* ; Microfluidics ; Plasma ; Ultracentrifugation

Lung cancer is a malignant tumor with high incidence and mortality in the world, which seriously threatens people's safety and health. Early diagnosis of lung cancer is the key part in the process of prevention and treatment of lung cancer. It can improve the survival of patients with lung cancer. Exosomes are closely related to the invasion and metastasis process of tumor, it plays an important role in the development of lung cancer. Biomarkers based on exosomes have become a powerful diagnostic tool of lung cancer. Exosomes are lipid bilayer vesicles with uniform size and diameter of 30 nm-200 nm secreted by cells. Exosomes contain different types of nucleic acids and proteins. These nucleic acids and proteins are derived from their parent cells (including parent cancer cells), which have a wide range of physiological functions, including immune regulation, intercellular communication and other physiological activities. Biomacromolecules in exosomes, such as single-stranded RNA, long noncoding RNA, microRNA, protein and lipids, which can provide valuable genetic information for early clinical diagnosis of lung cancer. This review described the origin, structural characteristics, extraction methods, biological characteristics of exosomes and the relationships of exosomes in the early diagnosis of lung cancer.
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Objective: To investigate the value of CT wavelet texture analysis based on primary tumor of papillary thyroid carcinoma (PTC) in predicting central lymph node metastasis (CLNM). Methods: A retrospective analysis was performed to 250 patients (307 nodules) who pathologically confirm with PTC in the First Affiliated Hospital of Kunming Medical University from December 2013 to August 2019. Thyroid dual phase scanning was performed in all patients within two weeks before surgery. All patients underwent central or total cervical lymph node dissection, among which 160 cases (189 nodules) were classified as training sets, while 90 cases (118 nodules) were in the verification sets. Besides, all patients were divided into CLNM group and no CLNM group according to pathology. The DeepWise software were used to manually delineate PTC primary nodules on venous phase CT images, and 576 wavelet texture features were extracted. The differences of texture feature parameters between the two groups were compared. The top 10 wavelet texture features of the area under curve (AUC) value were manually selected as the best parameters. Multivariable logistic regression analysis was used to establish and verify the model, the optimal cutoff value was found by using receiver operating characteristic curve analysis. Results: Totally 124 features were statistical difference between the two groups. The top 10 characteristic parameters for manual diagnosis with AUC values ranged from 0.599 to 0.630 (P<0.05), multi-collinearity test and multi-logstic regression analysis showed that there was no collinear correlation between the above 10 features, and small-area low-gray emphasis was an independent predictor of risk factors. The AUC value, sensitivity, specificity, and accuracy of the predictive model for the diagnosis of CLNM in the training set were 0.693, 62.84%, 60.47%, 62.96% and validation set were 0.602, 64.95%, 33.33%, and 59.32%, respectively. Conclusion Wavelet texture analysis in CT venous phase may allow detection of CLNM of PTC.

Respiratory management in patients with acute severe brain injury is an important part of the integrated critical management.A series of special pathophysiological changes and clinical manifestations after brain injury make it different from non-central nervous system diseases.Lung protective ventilation strategy and evidence from clinical trials have challenged the traditional view of respiratory management:hypocarbonemia and low PEEP strategy are no longer routine options for respiratory management in patients with brain injury,but respiratory management still needs to follow the principle of "avoiding secondary injury".Multimodal neuromonitoring shows good prospects,which is helpful to achieve the precision of respiratory management by optimizing the targets of the management of intracranial pressure,cerebral perfusion and cerebral oxygen metabolism.

"Adversity" is one primary element that impacts the pharmacology components of authentic Chinese herbal medicine. Knowledge about "adversity" is a precondition of yield estimation, quality monitoring, location selection and the geo-herbalism protection. Used 3S(GIS, RS and GPS) technology to combine multi-source key ecological factors of "Anling", and used parasitic relationships between organisms to extracting its suitable region for the first time. Results showed that the "Anling" were mainly distributed in Dabie coteau. Suitable area amount to 36.8 km², Yuexi, Shucheng, Jinzhai and Qianshan which account for about 93.55% of whole congenial region. The first three accounts for about 80.82%. It was Yuexi that account for 1/3 above especially. Field investigation verify accuracy of extraction about 91.67%, which has confirmed it is feasible that using the relationship between parasitic host and parasitic to extract native environment of parasitic traditional medicine based on 3S technology.

BACKGROUNDThymectomy is an established treatment for myasthenia gravis (MG), and video-assisted thoracoscopic surgery (VATS) thymectomy has become an acceptable surgical procedure. This study aimed to compare the results of VATS thymectomy and open thymectomy and to identify the prognostic factors after thymectomy.

METHODSThe clinical data of 187 consecutive thymectomies performed between July 2000 and December 2009 were retrospectively reviewed; 75 open thymectomies and 112 VATS thymectomies. Clinical efficacy and variables influencing outcome were assessed by Kaplan-Meier survival curves and Cox proportional hazards regression analysis.

RESULTSThe operative blood loss in the VATS group was significantly less than that in the open group ((62.14 ± 55.43) ml vs. (137.87 ± 165.25) ml, P < 0.05). The postoperative crisis rate increased with the severity of preoperative MG and the prescription dose of anticholinesterase. Complete follow-up information of patients more than 12 months after the thymectomy was obtained on 151 cases, 89 cases from the VATS group and 62 cases from the open group, with a mean follow-up period of 59.3 months, range from 12 to 117 months. Complete stable remission (CSR) was the end point for evaluation of the treatment results. The overall five-year CSR rate was 57.5%. Two good prognostic factors were identified; preoperative prescription of anticholinesterase alone (P = 0.035) and non-thymomatous MG (P = 0.003). The five-year CSR rate of the ocular type of MG reached a high level of 67.4%.

CONCLUSIONSThymectomy can achieve good long-term CSR in MG, and VATS is an ideal alternative method. High-dose prescription of anticholinesterase and the advanced stage by Myasthenia Gravis Foundation of America (MGFA) classification have higher risks of postoperative crisis. Preoperative prescription of anticholinesterase alone and non-thymomatous MG are good prognostic factors. Thymectomy should also be considered for the ocular type of MG.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; surgery ; Proportional Hazards Models ; Thoracic Surgery, Video-Assisted ; methods ; Thymectomy ; adverse effects ; methods ; Time Factors ; Treatment Outcome