Objective To investigate the impact of desmoglein 3 (Dsg3) on the proliferation of peripheral T lymphocytes from patients with pemphigus vulgaris (PV).Methods Peripheral blood mononuclear cells (PBMCs) were obtained from 12 patients with PV and 22 normal human controls,cultured with or without the presence of Dsg3 or phytohemagglutinin for 3 days.Flow cytometry was performed to detect the changes of T-lymphocyte subsets in PBMCs stimulated with Dsg3 and proliferation of T-lymphocytes.Results In patients with PV,the percentage of Th2 and Th1 cells was 12.17%±5.32% and 4.08%±1.50%,respectively in Dsg3stimulated PBMCs,9.84%±5.41% and 3.91%±1.38%,respectively in non-stimulated PBMCs.Increased percentage of Th2 cells was observed in Dsg3-stimulated and non-stimulated PBMCs from patients with PV compared with those from normal human controls (both P<0.05).After stimulation with Dsg3,there was a significant proliferation of T cells from patients,and the proliferation rate of CD4+T cells was 4.65%±3.28%,which was significantly higher than that from normal controls(P<0.05).Conclusion Dsg3 can induce the specific proliferation of CD4+T cells,especially Th2 type CD+4 T cells,from patients with PV.

Objective To observe the effects of fetal bovine serum(FBS) on differentiation of human neural stem cells (NSCs). Methods The effects of FBS with different concentrations on differentiation of human fetal NSCs were observed by cell culture, immunocytochemistry and flow cytometry. Results Human fetal NSCs could be induced to differentiate mainly three types of nerve system cells(neuron, astrocyte and oligodentrocyte). There were 80%～90% astrocytes of differentiated cells from human fetal NSCs with the concentration of 15% FBS induced. Conclusion Concentration dependent FBS in culture medium may have effect on the ratio of neurons to glial cells differentiated from human NSCs in vitro .

Objective To investigate whether IL-17 could stimulate the vascular endothelial growth factor (VEGF) production on HaCaT cells alone. We also investigated whether shikonin could inhibited the proinflamation effects of interleukin-17(IL-17) acting on HaCaT cells. MethodsWe examined the expression of VEGF by double antibody sandwich enzyme-linked immunosorbent assay ( ELISA ) and realtime polymerase chain reaction(RT-PCR) in HaCaT cells and the cell supernatant. The viability of HaCaT cells in the drug group was detected by the Cell Counting Kit-8 (CCK-8). ResultsThe expression of VEGF in different time IL-17-stimulated groups on HaCaT cells and the cell supernatant were higher than the control group( P＜0.001 ). The expression of VEGF in different drug treatment groups on HaCaT cells and the cell supematant were lower than the stimulated group by IL-17 ( P＜0. 001 ). The cell viability of different drug treatment groups have no significant difference( P＞0.05 ). ConclusionWe show that IL-17 specifically and time-dependently augmented and induced VEGF expression on HaCaT cells and the cell supernatantThen shikonin markedly inhibited the increase tengency of IL-17 effection on HaCaT cells and the cell supematant level.

OBJECTIVE: To observe and analyze the function and morphology of pharyngeal ostium of the eustachian tubes in secretory otitis media and chronic rhinosinusitis in children under direct vision,in order to provide an objective basis for clinical treatments. METHOD: Fifty cases of secretory otitis media,50 cases of chronic rhinosinusitis and a control group of 50 cases with hoarseness were examined under video laryngoscope to observe the pharyngeal ostium morphological changes of the eustachian tubes, and their functional statuses were tested by using acoustic impedance instrument. All the data were analyzed by statistical methods. RESULT: (1) In the secretory otitis group, the abnomal rate of the pharyngeal ostium of the eustachian tubes was 94% while the chronic rhinosinusitis group was 80%,and between them there was no significant differences (P > 0.05). But both of them had significant differences with the control group (P < 0.05). (2) In the secretory otitis group, the rate of the eustachian tube dysfunction was 70% while the chronic rhinosinusitis group was 26%, and between them there was significant differences (P < 0.05), and both of them have significant differences when compared with the control group (P < 0.05). CONCLUSION There are some abnormal points exist in the function and the morphology of the eustachian tube in secretory otitis media and chronic rhinosinusitis in children. Eustachian tube dysfunction played a dominant role in the pathogenesis of secretory otitis media in children rather than the morphological change did compared to the chronic rhinosinusitis in children.
Case-Control Studies ; Child ; Chronic Disease ; Eustachian Tube ; pathology ; physiopathology ; Humans ; Otitis Media with Effusion ; pathology ; physiopathology ; Rhinitis ; pathology ; physiopathology ; Sinusitis ; pathology ; physiopathology

Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.

Objective To analyze the MR imaging performances and pathological characteristics of nodular muscular sarcoidosis,and to improve the diagnostic accuracy.Methods Six cases with pathologically confirmed nodular muscular sarcoidosis were included,the MRI imaging findings and pathologic appearance were analyzed,and the literature was reviewed.Results Six patients underwent plain and enhanced MRI examination with 3 cases of unilateral gastrocnemius muscle single lesion,1 case of unilateral anterior medial thigh muscle lesions,and 2 cases of bilateral gastrocnemius muscle lesions.MRI plain scan showed a strip-shaped or fusiform with slightly short TI-long T2 signal along the muscle fibers,and the occupancy effect was not obvious."Dark star" sign showed in lesion center with axial T2WI and enhanced T1WI,and "three stripes" sign could be seen along the long axis view image lesions.The lesion obviously enhanced after enhancement,with mild enhancement center.Similar small nodular satellite lesions were seen around 5 large lesions,and 4 cases showed similar small nodule enhancement in distant muscle groups.Pathological manifestations:granulomatous inflammation of muscle fibers,intermuscular epithelial cells,and multinucleated tissue cells with or without lymphocytic infiltration;no cheese-like necrosis in the nodules,visible fusion of nodules.Conclusion On MRI,it is characterized by strips or fusiform lesions along the muscle fibers,visible "three-layer stripe sign" and "black star sign",and the characteristics of the enhanced muscle nodules in the surrounding muscle groups,which contributes to the diagnosis and differential diagnosis of muscular sarcoidosis.

Objective:To investigate the value of CT tumor-thyroid marginal contact range (MCR) for predicting capsular invasion and cervical lymph node metastasis in papillary thyroid carcinoma (PTC) with a diameter>1.0 cm and papillary thyroid microcarcinomas (PTMC) with a diameter ≤ 1.0 cm, and to evaluate the diagnostic efficacy of direct CT signs for lymph node metastasis.Methods:The CT data of 148 patients with PTC (>1.0 cm) and 193 patients with PTMC confirmed by surgery and pathology were retrospectively analyzed from January 2017 to April 2020 at Hangzhou First People′s Hospital. MCR was evaluated based on CT images and classified as<1/4 tumor circumference or ≥1/4 tumor circumference. Direct CT signs of cervical lymph nodes were observed, including cystic change, microcalcification, hyperenhancement, short/long diameter≥0.5, clustered lymph nodes or central area turbidity. The difference in the distribution of MCR between PTC (>1.0 cm) and PTMC was compared using the χ 2 test, and the efficiency of MCR and direct CT signs for diagnosing capsular invasion and lymph node metastasis was calculated using the pathological results as the gold standard. Results:In 148 PTC (>1.0 cm) and 193 PTMC patients, the pathological results showed capsular invasion in 88.5% (131/148) and 57.0% (110/193), and lymph node metastasis in 71.6% (106/148) and 44.0% (85/193), respectively. In PTC (>1.0 cm) patients, MCR≥1/4 tumor circumference was more common in patients with capsular invasion (χ2=22.211, P<0.001) and lymph node metastasis (χ2=4.746, P=0.029), and the corresponding sensitivity and specificity for predicting capsular invasion and lymph node metastasis were 84.7% (111/131) and 64.7% (11/17), 83.0% (88/106) and 33.3% (14/42), respectively. In PTMC patients, MCR≥1/4 tumor circumference was more common in patients with capsular invasion (χ2=66.066, P<0.001) and lymph node metastasis (χ2=5.343, P=0.021), and its sensitivity and specificity for predicting capsular invasion and lymph node metastasis were 87.3% (96/110) and 69.9% (58/83), 71.8% (61/85) and 44.4% (48/108), respectively. The sensitivity and specificity of direct CT signs for diagnosing lymph node metastasis in PTC (>1.0 cm) and PTMC were 89.6% and 73.8%, 69.4% and 76.9%, respectively. Conclusions:Both direct CT signs and MCR ≥ 1/4 tumor circumference can predict cervical lymph node metastasis in PTC patients, and the former had higher sensitivity and specificity. MCR≥1/4 tumor circumference has high efficiency for predicting capsular invasion in PTC patients.

Objective: To investigate the prognostic value of radiomics analysis in predicting axillary lymph nodes (ALN) metastasis of breast cancer based on dynamic contrast-enhanced MR imaging (DCE-MRI). Methods: One hundred and ninety-six patients with suspected breast cancer were prospectively collected for dynamic breast DCE-MRI. Enhanced MR imaging data of 72 axillary lymph nodes were evaluated separately by a chief radiologist and a resident, and the consistency analysis was performed. Lymph nodes were dichotomized according to the pathology results derived from operation or biopsy under real-time virtual sonography based on MRI data. Clinical and imaging data were also divided into corresponding groups. (Imaging) Data from both groups were respectively classified as training set and testing set by stratified sampling in proportion with 3∶1. AK software was applied to extract 6 major categories of 385 features (including histogram, morphology, texture parameters, gray level co-occurrence matrix, run-length matrix and grey level zone size matrix from imaging), and a set of statistically significant features were subsequently obtained by dimension reduction. The prediction model was established through binary classification logistic regression and employed to externally test the validation set by the method of confusion matrix. Meanwhile, ROC analysis was applied to assess the diagnostic performance of the model. Results: Of the 72 axillary lymph nodes, 35 were metastatic negative and 37 were positive. The consistency of enhanced MRI radiomics features was good, between 0.841 and 0.980. Uniformity, ClusterProminence_AllDirection_offset1_SD, Correlation_AllDirection_offset1, LongRunEmphasis_angle90_offset7 and SurfaceVolumeRatio were statistically significant differences (P<0.01), the area under the ROC between 0.747 and 0.931. In the training and testing group, the areas under the ROC, sensitivity, specificity and accuracy of the model were 0.953, 0.893, 0.926, 92.6% (50/54) and 0.944, 0.900, 1.000, 88.9% (16/18) respectively. Conclusion The prediction model based on radiomic features may provide a non-invasive and effective approach to the assessment of the risk of ALN metastasis of breast cancer.

We aimed to express and purify three rabies virus glycoproteins with different tags and sizes. After analyzing their binding function, we wish to obtain a rabies virus glycoprotein with higher affinity and ability to specifically bind memory B cells. Experiments were carried out to express full length, as well as the ectodomain RVG by gene engineering method. Combined with the antibody of CD19 and CD27, the candidate protein labeling with fluorescence was used to analyze its binding function. Flow cytometry was used to detect the anti-rabies virus specific memory B cells in PBMCs, and confirm the binding ability between the candidate proteins and anti-rabies virus-specific memory B cells. We successfully constructed three expression vectors pGEX-5X-1-RVG, pET28a-RVG and pET30a-G. Three glycoproteins GST-RVG, His-RVG and His-G were obtained by optimized expression and purification conditions. The antigen specificity of purified GST-RVG, His-RVG and His-G were identified by Western blotting and ELISA. The affinity of these three purified glycoproteins to anti-rabies virus antibody were detected by competitive ELISA. Anti-rabies virus specific memory B cells in positive PBMCs gained from people who had ever been injected with the vaccine can be detected by flow cytometry. Thus, we got a recombinant rabies virus glycoprotein that had high-affinity and could sort antigen specific memory B cells.

To scientifically evaluate the intervention effect of Chinese medicine preventive administration(combined use of Huo-xiang Zhengqi Oral Liquid and Jinhao Jiere Granules) on community population in the case of coronavirus disease 2019(COVID-19), a large cohort, prospective, randomized, and parallel-controlled clinical study was conducted. Total 22 065 subjects were included and randomly divided into 2 groups. The non-intervention group was given health guidance only, while the traditional Chinese medicine(TCM) intervention group was given two coordinated TCM in addition to health guidance. The medical instructions were as follows. Huoxiang Zhengqi Oral Liquid: oral before meals, 10 mL/time, 2 times/day, a course of 5 days. Jinhao Jiere Granules: dissolve in boiling water and take after meals, 8 g/time, 2 times/day, a course of 5 days, followed up for 14 days, respectively. The study found that with the intake of medication, the incidence rate of TCM intervention group was basically maintained at a low and continuous stable level(0.01%-0.02%), while the non-intervention group showed an overall trend of continuous growth(0.02%-0.18%) from 3 to 14 days. No suspected or confirmed COVID-19 case occurred in either group. There were 2 cases of colds in the TCM intervention group and 26 cases in the non-intervention group. The incidence of colds in the TCM intervention group was significantly lower(P<0.05) than that in the non-intervention group. In the population of 16-60 years old, the incidence rate of non-intervention and intervention groups were 0.01% and 0.25%, respectively. The difference of colds incidence between the two groups was statistically significant(P<0.05). In the population older than 60 years old, they were 0.04% and 0.21%, respectively. The incidence of colds in the non-intervention group was higher than that in the intervention group, but not reaching statistical difference. The protection rate of TCM for the whole population was 91.8%, especially for the population of age 16-60(95.0%). It was suggested that TCM intervention(combined use of Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules) could effectively protect community residents against respiratory diseases, such as colds, which was worthy of promotion in the community. In addition, in terms of safety, the incidence of adverse events and adverse reactions in the TCM intervention group was relatively low, which was basically consistent with the drug instructions.
Adolescent ; Adult ; Betacoronavirus ; Coronavirus Infections ; drug therapy ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional ; Middle Aged ; Pandemics ; Pneumonia, Viral ; drug therapy ; Prospective Studies ; Young Adult