An in vitro detection method of the gastrointestinal absorption of Pilose Antler protein was established for mixed protein activity. Five bands of protein with molecular weight of 17.8-160 kD derived from the Pilose Antler were extracted and sufficiently labeled with FITC (FITC-PE). The stability and variation of FITC-PE in gastrointestinal circumstances were detected by native polyacrylamide gel electrophoresis and confocal laser scanning microscope. Results showed that the main component of FITC-PE kept invariant after being reacted with artificial gastric fluid and artificial intestinal fluid. The fluorescence signal was detected 20 min after administration in the valgus intestinal purse experiment, and three kinds of protein, with molecular weight of 45, 25, and 17.8 kD, were detected in the mixture of absorbent protein. The research laid the foundation for the further in vivo study of Pilose Antler protein. Meanwhile, it would be an in vitro screening method for the absorption, distribution and metabolism of mixed protein from traditional Chinese medicine.

Objective To compare the cytology diagnostic accuracy of DNA quantitative cytology and thinprep cytology test(TCT) in cervical cancer screening for exploring effective method in cervical cancer screening.Methods TCT and DNA quantitative cytology were carried out in 7 470 women.Women with positive results additionally underwent high risk human papillomavirus (HPV) detection.Positive cytologic diagnosis included atypical squamous cells(ASC) or above in TCT and DNA index 2.5 or above in DNA quantitative cytology.Results The positive rate was 13.0％ in method of DNA quantitative cytology and 13.7％ in method of TCT in 7 470 cases.Positive rate of the two methods had no significant difference in cervical cancer screening(x2 =1.813,P =0.178).There was significant difference in positive rate of TCT between cases with DNA index≥2.5,＜4.5,heteroploid cells more than 3 or DNA index≥4.5 and cases with DNA index≥2.5,＜4.5,heteroploid cells less than 3.Every grade of TCT abnormality had abnormal DNA index.Abnormality of DNA index had an increasing trend with the severity of TCT.Infection rate of high risk HPVs had significant difference in different grades of DNA index (x2 =62.648,P =0.000).Conclusion Combination of DNA quantitative cytology and TCT is an effective method in cervical cancer screening,which can reduce misdiagnosis,guide cervical biopsy and suggest infection of high risk of HPVs.

To observe the effects of Huanglian Jiedu Decoction (HJD), a compound traditional Chinese herbal medicine, on lipid metabolism and its related gene expressions in rats with hyperlipidemia.

Objective:To observe and analyze the accuracy of the optic disc positioning and segmentation method of fundus images based on deep learning.Methods:The model training strategies were training and evaluating deep learning-based optic disc positioning and segmentation methods on the ORIGA dataset. A deep convolutional neural network (CNN) was built on the Caffe framework of deep learning. A sliding window was used to cut the original image of the ORIGA data set into many small pieces of pictures, and the deep CNN was used to determine whether each small piece of picture contained the complete disc structure, so as to find the area of the disc. In order to avoid the influence of blood vessels on the segmentation of the optic disc, the blood vessels in the optic disc area were removed before segmentation of the optic disc boundary. A deep network of optic disc segmentation based on image pixel classification was used to realize the segmentation of the optic disc of fundus images. The accuracy of the optic disc positioning and segmentation method was calculated based on deep learning of fundus images. Positioning accuracy=T/N, T represented the number of fundus images with correct optic disc positioning, and N represented the total number of fundus images used for positioning. The overlap error was used to compare the difference between the segmentation result of the optic disc and the actual boundary of the optic disc.Results:On the dataset from ORIGA, the accuracy of the optic disc localization can reach 99.6%, the average overlap error of optic disc segmentation was 7.1%. The calculation errors of the average cup-to-disk ratio for glaucoma images and normal images were 0.066 and 0.049, respectively. Disc segmentation of each image took an average of 10 ms.Conclusion:The algorithm can locate the disc area quickly and accurately, and can also segment the disc boundary more accurately.

Objective:To evaluate the efficiency of ResNet50-OC model based on deep learning for multiple classification of color fundus photographs.Methods:The proprietary dataset (PD) collected in July 2018 in BenQ Hospital of Nanjing Medical University and EyePACS dataset were included.The included images were classified into five types of high quality, underexposure, overexposure, blurred edges and lens flare according to clinical ophthalmologists.There were 1 000 images (800 from EyePACS and 200 from PD) for each type in the training dataset and 500 images (400 from EyePACS and 100 from PD) for each type in the testing dataset.There were 5 000 images in the training dataset and 2 500 images in the testing dataset.All images were normalized and augmented.The transfer learning method was used to initialize the parameters of the network model, on the basis of which the current mainstream deep learning classification networks (VGG, Inception-resnet-v2, ResNet, DenseNet) were compared.The optimal network ResNet50 with best accuracy and Micro F1 value was selected as the main network of the classification model in this study.In the training process, the One-Cycle strategy was introduced to accelerate the model convergence speed to obtain the optimal model ResNet50-OC.ResNet50-OC was applied to multi-class classification of fundus image quality.The accuracy and Micro F1 value of multi-classification of color fundus photographs by ResNet50 and ResNet50-OC were evaluated.Results:The multi-classification accuracy and Micro F1 values of color fundus photographs of ResNet50 were significantly higher than those of VGG, Inception-resnet-v2, ResNet34 and DenseNet.The accuracy of multi-classification of fundus photographs in the ResNet50-OC model was 98.77% after 15 rounds of training, which was higher than 98.76% of the ResNet50 model after 50 rounds of training.The Micro F1 value of multi-classification of retinal images in ResNet50-OC model was 98.78% after 15 rounds of training, which was the same as that of ResNet50 model after 50 rounds of training.Conclusions:The proposed ResNet50-OC model can be accurate and effective in the multi-classification of color fundus photograph quality.One-Cycle strategy can reduce the frequency of training and improve the classification efficiency.

Objective: To analyze the application of accelerated rehabilitation surgery in elderly patients with gastric cancer surgery and its influence on inflammation and nutritional indicators. Methods: From October 2017 to October 2018, 80 elderly patients with gastric cancer who underwent gastrectomy in Shaoxing People's Hospital were selected.According to random number table method, they were randomly divided into traditional control group and ERAS group, with 40 cases in each group.The traditional control group was treated by traditional perioperative treatment + operation, while ERAS group was treated with ERAS perioperative treatment + operation.The recovery and complications, inflammation and nutritional changes before operation, 1 day after operation and 3 days after operation, and the improvement of quality of life after operation were compared and analyzed between the two groups. Results: In the ERAS group, the first exhaust time[(2.3±0.8)d] and defecation time[(2.5±0.4)d]were shorter than those in the traditional control group[(3.5±0.5)d and (3.7±0.6)d], and the incidence rate of complications (7.5%) was lower than that in the traditional control group (35.0%), the differences were statistically significant (t=8.72, 10.52, χ²=9.04, all P<0.05). On the first day after operation, the serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6(IL-6) in the ERAS group were (28.3±4.9)μg/L and (23.1±5.8)μg/L, respectively, which were lower than those in the traditional control group[(39.8±6.7)μg/L and (34.5±8.1)μg/L](t=8.76, 7.24, all P<0.05). The serum TNF-α and IL-6 levels in the ERAS group on the 3rd day after operation were (18.4±6.2)μg/L and (18.3±3.2)μg/L, respectively, which were lower than those in the traditional control group[(26.2±5.1)μg/L and (26.5±4.8)μg/L](t=6.14, 8.99, all P<0.05). The serum levels of albumin (Alb) and prealbumin (PRE) in the ERAS group on the first day after operation were (31.1±0.7)g/L and (161.3±7.2) mg/L, respectively, which were lower than those in the traditional control group[(30.2±0.9)g/L and (154.3±4.8)mg/L](t=4.99, 5.12, all P<0.05). The serum levels of Alb and PRE in the ERAS group on the 3rd day after operation were (33.4±0.5)g/L and (172.1±5.0)mg/L, respectively, which were lower than those in the traditional control group[(31.9±0.8)g/L and (165.3±3.2)mg/L](t=10.06, 7.24, all P<0.05). The WHOQOL-BREF score of the ERAS group[(91.4±6.5)points]was higher than that of the traditional control group[(80.3±7.8)points](P<0.05). Conclusion Accelerated rehabilitation surgery has good effect for elderly gastric cancer surgery.It can maintain the inflammation and nutritional status of patients after operation.It is worthy of clinical reference.

Objective:To analyze the damage in hippocampal tissues of mice after whole-body irradiation with high- or low-dose ionizing radiation and to investigate the roles of microglia/macrophages polarization in the injury.Methods:C57BL/6 mice were randomly divided into three groups: sham irradiation group, low-dose group (0.05 Gy) and high-dose group (7 Gy). Low- and high-dose groups were respectively treated by whole-body irradiation with single dose of 60Co γ-rays. Hippocampal tissues of the mice were collected at 6 h, 1 d, 3 d and 7 d after irradiation. The morphology, structure and apoptosis of neurons were detected by HE staining, Nissl staining and Tunnel staining, respectively. RT-PCR and immunofluorescence assay were performed to detect the expression of M1 and M2 microglial markers at mRNA and protein levels in hippocampus tissues. The cognitive and emotional behaviors of mice were evaluated one month after the irradiation by Morris water maze, open field test, elevated plus maze and tail suspension test. Results:There were morphological and structural changes in the nerve cells in the hippocampus region of mice after irradiation, accompanied by apoptosis. Acute injuries occurred at 6 h after radiation, alleviated at 1 d and 3 d, and persisted at 7 d in a dose-dependent manner. The results of immunofluorescence staining and confocal imaging analysis showed that compared with the sham irradiation group, the high-dose group showed increased number of microglia, down-regulated expression of M1 microglial markers and up-regulated expression of M2 microglial markers in the hippocampus at 6 h and 1 d after radiation, while M2 microglial markers decreased at 3 d and 7 d after irradiation. PCR results showed that the expression of M1 and M2 microglial markers at mRNA level in the irradiation groups increased at 6 h after irradiation, but there was no statistical significance. The expression of related proinflammatory/anti-inflammatory factors was significantly up-regulated. The results of behavioral experiments showed that compared with the sham irradiation group, there was no statistical difference in cognitive or emotional behaviors at one month after irradiation.Conclusions:60Co γ-rays could damage mouse hippocampal tissues and result in the overexpression and different polarization patterns of microglia/macrophages in mice.

Objective: To investigate the clinical features of adult B-cell acute lymphoblastic leukemia (B-ALL) with E2A-PBX1 fusion gene positive. Methods: The clinical data of 91 Philadelphia chromosome negative B-ALL patients who received chemotherapy regularly in Nanfang Hospital of Southern Medical University from March 2015 to December 2017 were collected. According to the fusion gene detection results, the patients were divided into E2A-PBX1 fusion gene positive group and E2A-PBX1 fusion gene negative group. The clinical features and prognosis of both groups were retrospectively analyzed. And then E2A-PBX1 fusion gene negative group was further divided into MLL-AF4 fusion gene positive group and the other ALL groups,which were compared with E2A-PBX1 fusion gene positive group in the prognosis. Results: There were 11 ALL patients with E2A-PBX1 fusion gene positive, and 80 ALL patients with E2A-PBX1 fusion gene negative. The level of lactic dehydrogenase (LDH) in E2A-PBX1 fusion gene positive group was higher than that in E2A-PBX1 fusion gene negative group [4 063 U/L (1 070- 9 554 U/L) vs. 454 U/L (103- 18 651 U/L), U=-4.700, P < 0.01], and there were no statistically significant differences in age, gender, white blood cell count and extramedullary invasion (all P > 0.05). The immunophenotypes of ALL patients with E2A-PBX1 fusion gene positive were all common B-cell phenotype. Out 4 of 7 ALL patients showed the different expression of Philadelphia-like ALL phenotype CRLF2 or phosphorylated CRKL or phosphorylated STAT5 by using flow cytometry. The complete remission (CR), minimal residual disease (MRD)-negative and the recurrence for 11 cases of E2A-PBX1 fusion gene positive group included 9, 7, 8 cases, respectively after first course of induction chemotherapy. The CR, MRD-negative and the recurrence for 80 cases of E2A-PBX1 fusion gene negative group included 71, 53, 26 cases, respectively after the first course of induction chemotherapy. There were no statistically significant differences in the CR and MRD-negative between the two groups after the first course of induction chemotherapy (P= 0.721, P= 0.487), but there was a statistical difference in the recurrence (χ²= 7.751, P= 0.012). The median follow-up time was 17 months (1-44 months), and the 1-year overall survival (OS) rate and 1-year event-free survival (EFS) rate of E2A-PBX1 fusion gene positive ALL group were lower than those of the negative group (OS: 43.6% vs. 70.0%, P= 0.020; EFS: 13.6% vs. 60.0%, P= 0.002). Subgroup analysis showed that there were no statistical differences between E2A-PBX1 fusion gene positive group and MLL-AF4 fusion gene positive group in 1-year OS rate and 1-year EFS rate (P= 0.617, P= 0.984). Conclusions E2A-PBX1 fusion gene positive rate is low in adult B-ALL patients with a good response to traditional chemotherapy in the early stage, but its cumulative recurrence rate is high. It is a high-risk cytogenetic abnormality which is similar to MLL gene rearrangement and needs to explore novel therapy strategies.

Objective:To detect the status of PIK3CA in triple-negative breast cancer (TNBC) , and to analyze the relationships between PIK3CA mutation and clinical features and its impact on prognosis.Methods:From January 1, 2016 to December 31, 2018, 50 patients with primary TNBC admitted to Xinxiang Central Hospital of Henan Province were collected. The PIK3CA mutation status was detected, and the relationships between PIK3CA mutation and clinical characteristics of patients with TNBC and its impact on prognosis were analyzed.Results:PIK3CA gene mutation was detected in 9 of 50 TNBC patients, with a mutation frequency of 18.0%. H1047R mutation was found in 4 cases, E545K mutation in 3 cases and E542K mutation in 2 cases. PIK3CA gene mutation was not associated with age ( χ2=3.55, P=0.060) , tumor location ( χ2=1.01, P=0.315) , tumor size ( χ2<0.01, P>0.999) , lymph node status ( χ2=0.76, P=0.385) , clinical stage ( χ2=0.65, P=0.420) , Ki-67 value ( χ2<0.01, P>0.999) , P53 status ( χ2=0.02, P=0.894) and human epidermal growth factor receptor-2 (HER-2) status ( χ2=1.65, P=0.200) . Prognostic analysis showed that 3-year disease-free survival rates of wild-type PIK3CA patients was significantly higher than that of mutant PIK3CA patients (80.5% vs. 11.1%, χ2=28.23, P<0.001) . Conclusion:The frequency of PIK3CA gene mutation is higher in TNBC patients. There is no correlation between PIK3CA mutation and clinicopathologic features in TNBC patients. PIK3CA gene mutation may be significantly associated with poor prognosis of TNBC patients.

Objective:To analyze the relationship between hematological and genotype characteristics of fetuses and patients with hemoglobin (Hb) H disease and their natural disease progression.Methods:From 2010 to 2022, a total of 1 252 blood samples from fetuses and patients with Hb H disease who visited the Guangxi Zhuang Autonomous Regional Maternal and Child Health Hospital were collected (including 174 umbilical cord blood samples, 1 062 peripheral blood samples from patients over 2 years old, and 16 peripheral blood samples from patients with rare cases of genotype Hb H). Additionally, 278 peripheral blood samples were collected from patients aged 0 - 2 years old with Hb H 3.7, Hb H 4.2, Hb H CS, and Hb H WS disease for the study of trends in red blood cell development. Multiple probe hybridization and microarray comparative genomic hybridization technology combined with first-generation Sanger sequencing were used for rare mutation detection.Results:Among the 1 062 Hb H disease patients over 2 years old, 62.34% (662/1 062) had gene deletion (--/-α), of which Hb H 3.7 (-- SEA/-α 3.7) and Hb H 4.2 (-- SEA/-α 4.2) were the most common, accounting for 42.28% (449/1 062) and 19.11% (203/1 062) of the total, respectively. Among the non-deletion genotypes (--/αα T or α Tα/αα T), Hb H CS (-- SEA/α CS), Hb H WS (-- SEA/α WS) and α CSα/α CSα accounted for 16.85% (179/1 062), 16.48% (175/1 062) and 1.98% (21/1 062), respectively. The 81.12% (537/662) of patients with deletional Hb H disease showed mild to moderate anemia, with Hb H detection rates ranging from 75% to 80%. Among non-deletional Hb H disease, Hb H WS disease showed the mild (blood Hb concentration > 95 g/L in 90% of patients) phenotype while Hb H CS and Hb H QS (-- SEA/αα QS) patients had moderate to severe anemia, with Hb H detected in peripheral blood at higher levels than in other types of Hb H disease patients. Except for Hb H CS and Hb H QS, which did not show a significant increase in Hb A2 levels when complicated with β-thalassemia, Hb A2 levels were increased (> 3.5%) in all other types of Hb H disease patients. When Hb H disease was complicated with β-thalassemia, Hb H peaks were not detected in either type of Hb H disease. The results of red blood cell development trend detection showed that erythrocyte counts were elevated in patients with Hb H disease compared to their normal counterparts; whereas, blood Hb, mean erythrocyte volume (MCV) and mean erythrocyte hemoglobin content (MCH) were lower than in their normal counterparts ( P < 0.05) and decreased to the minimum at 6 months to 1 year of age. Patients with Hb H CS disease, as the most severe form of anemia, had the highest MCV values ( P < 0.001). The results of fetal cord blood with Hb H disease showed that α CSα/α CSα caused severe intrauterine anemia, followed by Hb H QS and Hb H CS. The content of Hb Bart's in umbilical cord blood was negatively correlated with the severity of anemia ( rs = - 0.58, P < 0.001). When Hb H disease was complicated with β-thalassemia, there was no significant improvement in fetal anemia, and the Hb Bart's content did not change significantly ( P > 0.05). In addition, Hb H 21.9 (-α 21.9kb/-- SEA) and Hb H 2.4 (-α 2.4/-- SEA) were common in patients with deletion rare Hb H. In patients with non-deletion rare Hb H, αα Amsterdam-A1/-- SEA and αα Hb G-Georgia/-- SEA were both first reported. Conclusions:There is heterogeneity in clinical manifestations of patients with different types of Hb H disease or same type of Hb H disease at different developmental stages. When patients with Hb H are complicated with β-thalassemia, the phenotype of patients with the deletion type is improved, while that of patients with the non-deletion type is not. Compared to normal individuals, patients with Hb H disease have lower blood Hb concentration, MCV and MCH, and more rapid physiological changes in red blood cells.