Objective To compare the efficacy of five Blastocystis staining method for staining cells with different morphologies and in different developmental stages to select appropriate staining techniques for specific scenarios and experimental purposes. Methods Blastocystis was stained using Wright-Giemsa, periodic acid-Schiff (PAS), Oil Red O, Nile Red, and Lugol's iodine solution respectively. The staining effects of the five method on Blastocystis were compared. Results Wright-Giemsa staining colored the cytoplasm and nucleus of Blastocystis bluish-purple, with other parts lightly stained; PAS staining rendered Blastocystis cells red; Oil Red O staining showed different structures within Blastocystis in varying intensities of orange-red; Nile Red enabled Blastocystis to fluoresce orange-yellow under a fluorescence microscope; Lugol's iodine solution stained the cytoplasm and nucleus a dark brownish-yellow. Conclusions Wright-Giemsa staining is recommended for clinical diagnosis. For metabolic studies, a combination of periodic acid-Schiff (PAS) staining and Oil Red O staining is suggested; PAS + Oil Red O staining can provide comprehensive morphological evidence for cell metabolism research. Nile Red staining is recommended for dynamic observation of live Blastocystis. Lugol's iodine staining is suitable for preliminary screening of large-scale samples.

BACKGROUND:Currently,there have been studies on three-dimensional digitalization and visualization systems for adult acupoints,but there are not many reports on the visualization of pediatric acupoints based on real pediatric digital sectional anatomical datasets. OBJECTIVE:To design and develop a digital three-dimensional visualization system for children's neck acupoints,to provide a basis for acupuncture and moxibustion,meridian and acupoint science teaching,clinical practice,acupuncture manipulation practice,and acupuncture safety research,and to provide a basis for the development of children's acupoint simulation system. METHODS:Based on a real cross-sectional anatomical dataset of pre-school boys,a three-dimensional digital virtual anatomical model of the neck region of children and internal multi-organ three-dimensional reconstruction were completed using PhotoShop 2021 and Digihuman Reconstruction System software.A database of 11 acupoints was compiled,including Fengfu and Fengchi,using the Unity database language.A three-dimensional model of children's neck anatomy,acupoint database,and writing acupuncture operation codes were integrated in Unity3D software.A three-dimensional digital visualization system for children's neck acupoints was successfully created,which integrated simulation acupoint positioning,three-dimensional acupoint anatomy,acupuncture training,clinical teaching,and acupuncture safety research. RESULTS AND CONCLUSION:(1)This study was based on real child specimens.Manual layer by layer segmentation of cross-sectional images was used to ensure the accuracy of the three-dimensional model to the greatest extent possible.The 3D software Digihuman Reconstruction System was utilized to extract and save independent segmentation data.PhotoShop 2021 software was collaborated with to complete dozens of three-dimensional reconstruction anatomical models of the outer skin of the neck and its internal bone structure,cervical spinal cord,blood vessels and nerves,muscles,and ligaments in children.The basic morphology and overall contour integrity verification of each independent structure were completed in MeshLab software.The 3-material research 13.0 software was applied for final fine tuning and anatomical position confirmation,successfully simulating and restoring the true anatomical morphology of the neck of preschool children.(2)Based on and referring to the national standards of the People's Republic of China,a database of commonly used acupoints in children's neck region was collected and organized,including their names,meridians,positioning,local anatomy,needle insertion levels,acupuncture methods,acupuncture accidents and prevention,acupoint indications,and two-dimensional anatomical sectional images.(3)Unity3D software was employed to integrate the three-dimensional model of children's neck,acupuncture simulation operation,and acupoint database,and a three-dimensional digital children's neck acupoint acupuncture visualization system was successfully constructed.The system displayed information on children's neck acupoints,two-dimensional and three-dimensional anatomical structures,and achieved two-dimensional and three-dimensional acupuncture simulation functions and acupuncture safety research functions for children's neck acupoints.Based on the ultra-thin sectional anatomical dataset of real child specimens,the first three-dimensional digital and visualization system for acupoints in the neck region of children had been constructed.Compared with previous acupoint acupuncture systems,it is more in line with the anatomical and morphological development characteristics of Asian children and has high application value in the fields of acupuncture safety research,clinical teaching,and acupuncture simulation training.

Objectives:To investigate the relationship between serum miR-550a-5p levels and spinal metas-tasis in patients with non-small cell lung cancer(NSCLC).Methods:The clinical data of 175 patients with NSCLC who received treatment in the Affiliated Hospital of Chengde Medical College between May 2021 and May 2023 were retrospectively analyzed.According to the results of whole body bone imaging,70 patients with spinal metastasis were enrolled into the metastasis group,and 104 patients without spinal metastasis were enrolled into the none-metastasis group.There were no statistically significant differences in gender,age,smoking,hypertension,thrombosis,and tumor location between the two groups(P＞0.05).The pathological type,clinical stage,lymph node metastasis,and tumor diameter of the patients were collected and compared between the two groups.qRT-PCR was used to detect the expression level of serum miR-550a-5p,enzyme-linked immunosorbent assay was used to detect the serum tartrate-resistant acid phosphatase-5b(TRACP-5b)and type Ⅰ collagen cross-linked C-terminal peptide(ICTP)levels.The differences in serum levels of TRACP-5b,ICTP,and miR-550a-5p were compared between the two groups.The receiver operating characteristic(ROC)curve was adopted to evaluate the diagnostic value of miR-550a-5p,TRACP-5b,and ICTP alone and in combination for spinal metastasis in NSCLC patients.Results:Compared with the none-metastasis group,serum TRACP-5b,ICTP,and miR-550a-5p levels were significantly higher in patients in the metastasis group(P＜0.05).The miR-550a-5p level was significantly higher in patients with tumor diameter≥5cm,lung adenocarcinoma,stage Ⅲ-Ⅳ,and lymph node metastasis than in patients with tumor diameter＜5cm,squamous carcinoma,stage Ⅰ-Ⅱ,and no lymph node metastasis(P＜0.05).Elevated serum miR-550a-5p,lung adenocarcinoma,stage Ⅲ-Ⅳ,and lymph node metastasis were the independent risk factors for the development of spinal metastasis in NSCLC patients(P＜0.05).The AUC of miR-550a-5p for diagnosing spinal metastasis in NSCLC patients was 0.851,and the sensitivity and specificity were 74.29％and 85.58％,respectively,at the optimal cut-off value of 0.20.The diagnostic efficacy of serum miR-550a-5p for diagnosing spinal metastasis in NSCLC patients was significantly better than that of TRACP-5b(Z=2.309,P=0.023)and ICTP(Z=1.852,P=0.049).The AUC of miR-550a-5p,TRACP-5b,and ICTP combination for the diagnosis of spinal metastasis in NSCLC patients was 0.931,and the sensitivity and specificity were 88.57％and 87.50％,respectively.The diagnostic efficacy of the three-marker combination was significantly better than that of the two-marker combination of TRACP-5b and ICTP(Z=2.205,P=0.027).Conclusions:Elevated level of serum miR-550a-5p is associated with spinal metastasis,which can be used as a molecular marker for predicting spinal metastasis in NSCLC patients,moreover,its combination use with bone metabolism-related tumor markers,the ICTP and TRACP-5b,can improve diagnostic efficacy.

Objectives:To investigate the relationship between serum miR-550a-5p levels and spinal metas-tasis in patients with non-small cell lung cancer(NSCLC).Methods:The clinical data of 175 patients with NSCLC who received treatment in the Affiliated Hospital of Chengde Medical College between May 2021 and May 2023 were retrospectively analyzed.According to the results of whole body bone imaging,70 patients with spinal metastasis were enrolled into the metastasis group,and 104 patients without spinal metastasis were enrolled into the none-metastasis group.There were no statistically significant differences in gender,age,smoking,hypertension,thrombosis,and tumor location between the two groups(P＞0.05).The pathological type,clinical stage,lymph node metastasis,and tumor diameter of the patients were collected and compared between the two groups.qRT-PCR was used to detect the expression level of serum miR-550a-5p,enzyme-linked immunosorbent assay was used to detect the serum tartrate-resistant acid phosphatase-5b(TRACP-5b)and type Ⅰ collagen cross-linked C-terminal peptide(ICTP)levels.The differences in serum levels of TRACP-5b,ICTP,and miR-550a-5p were compared between the two groups.The receiver operating characteristic(ROC)curve was adopted to evaluate the diagnostic value of miR-550a-5p,TRACP-5b,and ICTP alone and in combination for spinal metastasis in NSCLC patients.Results:Compared with the none-metastasis group,serum TRACP-5b,ICTP,and miR-550a-5p levels were significantly higher in patients in the metastasis group(P＜0.05).The miR-550a-5p level was significantly higher in patients with tumor diameter≥5cm,lung adenocarcinoma,stage Ⅲ-Ⅳ,and lymph node metastasis than in patients with tumor diameter＜5cm,squamous carcinoma,stage Ⅰ-Ⅱ,and no lymph node metastasis(P＜0.05).Elevated serum miR-550a-5p,lung adenocarcinoma,stage Ⅲ-Ⅳ,and lymph node metastasis were the independent risk factors for the development of spinal metastasis in NSCLC patients(P＜0.05).The AUC of miR-550a-5p for diagnosing spinal metastasis in NSCLC patients was 0.851,and the sensitivity and specificity were 74.29％and 85.58％,respectively,at the optimal cut-off value of 0.20.The diagnostic efficacy of serum miR-550a-5p for diagnosing spinal metastasis in NSCLC patients was significantly better than that of TRACP-5b(Z=2.309,P=0.023)and ICTP(Z=1.852,P=0.049).The AUC of miR-550a-5p,TRACP-5b,and ICTP combination for the diagnosis of spinal metastasis in NSCLC patients was 0.931,and the sensitivity and specificity were 88.57％and 87.50％,respectively.The diagnostic efficacy of the three-marker combination was significantly better than that of the two-marker combination of TRACP-5b and ICTP(Z=2.205,P=0.027).Conclusions:Elevated level of serum miR-550a-5p is associated with spinal metastasis,which can be used as a molecular marker for predicting spinal metastasis in NSCLC patients,moreover,its combination use with bone metabolism-related tumor markers,the ICTP and TRACP-5b,can improve diagnostic efficacy.

Objective: To investigate the mechanism of signal transducer and activator of transcription 3 (STAT3) and cancer associated fibroblasts (CAF) jointly generate chemo-resistance in epithelial-ovarian cancer and their effect on prognosis. Methods: A total of 119 patients with high-grade ovarian serous cancer who received surgery in Cancer Hospital of Chinese Academy of Medical Sciences from September 2009 to October 2017 were collected. The clinico-pathological data and follow-up data were complete. Multivariate Cox regression model was used to analyze the prognostic factors. Ovarian cancer tissue chips of patients in our hospital were prepared. EnVision two-step method immunohistochemistry was used to detect the protein expression levels of STAT3, the specific markers of CAF activation, fibroblast activating protein (FAP), and type Ⅰ collagen (COL1A1) secreted by CAF. The relationship between the expression of STAT3, FAP, COL1A1 protein and drug resistance and prognosis of ovarian cancer patients was analyzed, and the correlation between the expression of three proteins was analyzed. These results were verified through the gene expression and prognostic information of human ovarian cancer tissues collected in the GSE26712 dataset of gene expression omnibus (GEO) database. Results: (1) Multivariate Cox regression model analysis showed that chemotherapy resistance was an independent risk factor for overall survival (OS) of ovarian cancer (P<0.001). (2) The expression levels of STAT3, FAP, and COL1A1 proteins in chemotherapy resistant patients were significantly higher than those in chemotherapy sensitive patients (all P<0.05). Patients with high expression of STAT3, FAP, and COL1A1 had significantly shorter OS than those with low expression (all P<0.05). According to the human ovarian cancer GSE26712 dataset of GEO database, patients with high expression of STAT3, FAP, and COL1A1 also showed shorter OS than patients with low expression (all P<0.05), the verification results were consistent with the detection results of ovarian cancer patients in our hospital. (3) Correlation analysis showed that the protein level of STAT3 was positively correlated with FAP and COL1A1 in our hospital's ovarian cancer tissue chips (r=0.47, P<0.001; r=0.30, P=0.006), the analysis of GEO database GSE26712 dataset showed that the expression of STAT3 gene and FAP, COL1A1 gene were also significantly positively correlated (r=0.31, P<0.001; r=0.52, P<0.001). Conclusion: STAT3 and CAF could promote chemotherapy resistance of ovarian cancer and lead to poor prognosis.
Female ; Humans ; Cancer-Associated Fibroblasts/pathology* ; Carcinoma, Ovarian Epithelial ; Ovarian Neoplasms/pathology* ; Prognosis ; STAT3 Transcription Factor/metabolism* ; Drug Resistance, Neoplasm

Objective: To investigate the clinical features, morphological characteristics, immunophenotype, and differential diagnosis of goblet cell adenocarcinoma (GCA) in the digestive system. Methods: The clinicopathological data, morphological characteristics, immunophenotypes of 22 cases of GCA in the digestive system diagnosed from January 2010 to January 2021 were collected. Meanwhile, 25 cases of neuroendocrine neoplasm (NEN) and 24 cases of adenocarcinoma were used as controls. Relevant literature was also reviewed. Results: There were 16 males and 6 females, aged from 36 to 79 years with an average of 56 years. The anatomical sites of the 22 GCA were mostly appendix (17 cases) and occasionally extra-appendix (5 cases), including 3 cases in stomach, 1 case in duodenum and 1 case in anal. All 17 cases of appendiceal GCA were pure GCA. Among the 5 cases of extra-appendiceal GCA, One case of gastric GCA was pure, two cases of gastric GCA with NEN or adenocarcinoma, duodenal GCA with NEN and adenocarcinoma, anal GCA with NEN.Low-grade GCAs were composed of goblet, Paneth and neuroendocrine cells, which were arranged in intestinal crypt tubular or cluster structures and distributed in the wall of digestive system. The tubular and cluster structures lacked adhesion. Goblet cells were columnar, located in the base, with clear cytoplasm, small nuclei, inconspicuous atypia, and uncommon mitoses. Extracellular mucus and signet-ring cells with nuclear variations could be seen in some cases. Nerve fiber bundle invasion and tumor thrombus in vessels were often present. High-grade GCAs lacked tubular and cluster structures, and their histological structures were more complex. Tumor cells expressed mixed neuroendocrine and glandular epithelial markers. Similar to the expression patterns of synaptophysin and chromogranin A, CD200 and INSM1 were also dot-like or patch-positive in GCA. Conclusions: GCA is an infrequent tumor of the digestive system and shows the bi-directional differentiation characteristics of neuroendocrine and glandular epithelium. Accurate diagnosis and staging are related to its prognosis.
Adenocarcinoma/pathology* ; Appendiceal Neoplasms/surgery* ; Carcinoid Tumor/surgery* ; Chromogranin A ; Female ; Goblet Cells/pathology* ; Humans ; Male ; Neuroendocrine Tumors/pathology* ; Repressor Proteins ; Synaptophysin

This study was designed to determine the metabolites of Zhali Nusi Prescription(ZLNSP) in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric(UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of ZLNSP in rat plasma, bile, urine and feces. The biological samples were analyzed by ACQUITY UPLC BEH T_3 column(2.1 mm×100 mm,1.7 μm), with 0.1% formic acid water(A)-acetonitrile(B) as mobile phase, and the biological samples were analyzed in negative ion mode by electrospray ionization mass spectrometry(ESI-MS). An analytical method for biological samples of rats was established, and 8 prototype components and 36 metabolites were identified. The results showed that the metabolic pathways of the main components of ZLNSP in rats included methylation, glucuronidation, sulfation and so on. It provi-ded information for the therapeutic effect of ZLNSP in vivo.
Administration, Oral ; Animals ; Bile ; Chromatography, High Pressure Liquid ; Feces ; Plasma ; Prescriptions ; Rats

OBJECTIVE: To summarize the experiences and outcomes of 108 robot-assisted laparoscopic upper urinary tract reconstruction surgeries conducted by a single surgeon. METHODS: We consecutively and retrospectively reviewed 108 patients who underwent robot-assisted laparoscopic upper urinary tract reconstruction surgeries by a single surgeon from November 2018 to January 2020. The patient demographics, perioperative variables, postoperative complications and follow-up data were recorded. Fifty-three modified dismembered pyeloplasties (MDP), 11 spiral flap pyeloplasties (SFP), 11 ure-teroureterostomies (UUT), 4 lingual mucosal onlay graft ureteroplasties (LMU), 5 appendiceal onlay flap ureteroplasties (AU), 11 ureteral reimplantations (UR), 6 Boari flap-Psoas hitch surgeries (BPS) and 7 ileal ureter replacements (IUR) were enrolled finally. The success was defined as the improvement in subjective pain levels, and the improvement in the degree of hydronephrosis at ultrasound. RESULTS: All the surgeries were successfully completed without open or laparoscopic conversion. The median operative time was 141 min (range: 74-368 min), median blood loss was 20 mL (range: 10-350 mL) and median hospital stay was 4 d (range: 3-19 d) in MDP group, with the success rate of 94.3%. The median operative time was 159 min (range: 110-222 min), median blood loss was 50 mL (range: 20-150 mL) and median hospital stay was 5 d (range: 3-8 d) in SFP group, with the success rate of 100%. The median operative time was 126 min (range: 76-160 d), median blood loss was 20 mL (range: 10-50 mL) and median hospital stay was 5 d (range: 4-9 d) in UUT group, with the success rate of 100%. The median operative time was 204 min (range: 154-250 min), median blood loss was 30 mL (range: 10-100 mL) and median hospital stay was 6 d (range: 4-7 d) in LMU group, with the success rate of 100%. The median operative time was 164 min (range: 135-211 min), median blood loss was 75 mL (range: 50-200 mL) and median hospital stay was 8.5 d (range: 6-12 d) in AU group, with the success rate of 100%. The median operative time was 149 min (range: 100-218 min), median blood loss was 20 mL (range: 10-50 mL) and median hospital stay was 7 d (range: 5-10 d) in UR group, with the success rate of 90.9%. The median operative time was 166 min (range: 137-205 min), median blood loss was 45 mL (range: 20-100 mL) and median hospital stay was 5 d (range: 4-41 d) in BPS group, with the success rate of 83.3%. The median operative time was 270 min (range: 227-335 min), median blood loss was 100 mL (range: 10-100 mL) and median hospital stay was 7 d (range: 5-26 d) in IUR group, with the success rate of 85.7%. CONCLUSIONS The surgeon performed and modified numerous complicated upper urinary tract reconstruction surgeries by the robotic platform, which facilitated the development of the standardized upper urinary tract reconstruction surgical technique.
Humans ; Laparoscopy ; Retrospective Studies ; Robotic Surgical Procedures ; Surgeons ; Treatment Outcome ; Ureter

This study aims to compare the differences of Paeonia lactiflora from different habitats by establishing fingerprint. The fingerprint of P. lactiflora was established by UPLC. The samples collected from Sichuan,Hebei,Henan,Shanxi and Anhui were analyzed. The common peaks were identified by UPLC-Q-TOF/MS. The relative peak area of the common peaks was analyzed through similarity evaluation system( 2012 edition) for chromatographic fingerprint of traditional Chinese medicine developed by the National Pharmacopoeia Commission. Twelve common peaks were obtained and ten components were identified by reference substance and literature comparison. The similarity of each sample to the reference fingerprint is greater than 0. 900. However,all samples were clearly divided into 5 groups according to habitats after PLS-DA analysis. The peaks 2,6( ethyl gallate),10( galloypaeoniflorin) and 12( benzoyl paeoniflorin) were found to be the main difference components between the samples from five different habitats through the VIP value map. The study found that the variety of ingredients in the different areas are basically similar. But there are some differences in the content of the four components. The results of this study can provide reference at choosing and utilizing P. lactiflora from different places comprehensively.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Ecosystem ; Paeonia ; chemistry ; Phytochemicals ; analysis ; Plant Roots ; chemistry

Day care practice at children′s specialist hospitals can improve the quality of care for children patients.The hospital adopted such means as improving the service management and regulations, the service process of day care, creating the " green channel" , and moving forward and extending the care service. Their practice proved that the day care mode can significantly shorten the waiting time for hospitalization, cut back medical costs, improve the medical experience of children and their family, and achieve all-win for " community-hospital-patient".